bluuebunny/crossref_metadata_2025 huggingface dataset:
Overexpression of RhoA enhances peritoneal dissemination: RhoA suppression with Lovastatin may be useful for ovarian cancer
Authors: Horiuchi Akiko , Kikuchi Norihiko , Osada Ryosuke , Wang Cuiju , Hayashi Akiko , Nikaido Toshio , Konishi Ikuo
Published: November 2008

Small guanosine triphosphatase RhoA has been known to re‐organize cytoskeletons and regulate cell migration. The present authors have previously reported that expression of RhoA is significantly increased in advanced ovarian carcinomas and also in the peritoneal disseminated lesions. The present study investigated whether overexpression of RhoA could alter the progressive behavior of ovarian cancer cells. The effect of various Rho inhibitors on the biological behavior of ovarian cancer cells in vitro and in vivo was also examined. A stable RhoA‐transfectant of an ovarian cancer cell line SKOV3 was generated and examined in vitro for alterations of proliferative activity and invasiveness, and also in the nude mice model for peritoneal dissemination. In addition, the effect of a specific Rho inhibitor (C3 exoenzyme), Rho kinase inhibitor (Y27632) and hydroxymethylglutaryl coenzyme A (HMG–CoA) reductase inhibitor (Lovastatin and Pravastatin) were studied in vitro and in vivo. Forced overexpression of RhoA did not alter proliferative activity but significantly increased the invasiveness in vitro, which was suppressed by addition of C3 exoenzyme, Y27834, Lovastatin and Pravastatin. In the nude mice model, the frequency of dissemination and the number of disseminated lesions were significantly increased in the RhoA transfectant than in the control. In addition, oral administration of Lovastatin significantly decreased the number of metastatic sites compared with the control. These findings suggest that upregulation and/or activation of RhoA play an important role in the peritoneal dissemination of ovarian carcinoma, and that Lovastatin might be a candidate for the possible, novel treatment for ovarian carcinoma patients with peritoneal dissemination. (Cancer Sci 2008; 99: 2532–2539)
Quality and reporting of trial design in scientific papers in Anaesthesia over 25 years
Authors: Langford R. A. , Huang G. H. , Leslie K.
Published: November 2008

We determined how the quality of trial design and its reporting in scientific papers published in Anaesthesia has changed in the last 25 years. All articles between the years 1983–87 and 2003–07 were reviewed and classified according to methodology. Reporting and trial design of all prospective, comparative clinical interventional trials were compared between the two time periods using 12 criteria. Fewer articles now originate from the United Kingdom and Ireland than 25 years ago. Although fewer human interventional trials are now published in Anaesthesia, the quality of these trials has improved in terms of study design, bias control and proper disclosure. Significant improvements were observed in all criteria of trial design except for the declaration of non‐primary adverse outcomes and the minimisation of the risk of type I errors. Further improvements could still be made with respect to sample size calculation, description of the method of randomisation, and blinding.
Expression of DLK1 splice variants during porcine adipocyte development in vitro and in vivo
Authors: Samulin J. , Berg P. R. , Sundvold H. , Grindflek E. , Lien S.
Published: November 2008

Delta‐like 1 (DLK1) belongs to the epidermal growth factor‐like transmembrane protein family and is involved in the regulation of adipogenesis. Several splice variants of DLK1 have been identified in various species, of which two have been previously identified in pig. Here, we present two novel porcine DLK1 splice variants DLK1A and DLK1C. The gene expression profile of these variants together with the previously described DLK1B and DLK1C2 variants was studied in adipose tissue depots of pigs and during adipocyte differentiation in vitro. The short DLK1C and DLK1C2 transcripts were most abundantly expressed and their expression was reduced during porcine adipogenesis.
Relationship between HIV protease inhibitors and QTc interval duration in HIV‐infected patients: a cross‐sectional study
Authors: Charbit Beny , Rosier Arnaud , Bollens Diane , Boccara Franck , Boelle Pierre‐Yves , Koubaa Afef , Girard Pierre‐Marie , Funck‐Brentano Christian
Published: November 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• The risks of torsade de pointes and QTc prolongation in HIV patients have been reported.

• Authors have shown that four protease inhibitors (PIs) blocked human ether‐a‐go‐go‐related gene (hERG), the current that underlies QTc prolongation and drug‐induced proarrhythmia, and have also reported cases of torsades and suggested that PIs were responsible for these adverse events.

• This earlier paper has had a major impact on the perception of the arrhythmogenic risk associated with PIs, and regulatory agencies in Europe and the USA have modified the labelling of PIs accordingly.

WHAT THIS STUDY ADDS

• The present study provides alternative explanations for QTc prolongation in a cohort of HIV patients and reports ECG abnormalities found in such patients.

• It does not confirm that PIs play a significant role in QTc interval prolongation in HIV patients.

• In contrast, it shows that QTc prolongation is related to common causes and to the duration of HIV infection rather than to anti‐HIV treatments.

AIMS

QTc interval prolongation and torsades de pointes have been reported in HIV‐infected patients. Protease inhibitors (PIs) are suspected to contribute to this adverse reaction. However, many factors can prolong QTc interval. We examined factors influencing QTc duration in HIV‐infected patients.

METHODS

Unselected HIV‐infected patients (n = 978) were enrolled in this prospective, single‐centre cross‐sectional study. Variables related to infection and treatments were collected. A digital electrocardiographic record was recorded in each patient and QT interval duration was measured and corrected using both Bazett's (QTcB) and Fridericia's (QTcF) formula. Results were analysed with a multivariable linear model.

RESULTS

After excluding arrhythmias and complete bundle branch blocks, QT interval was measured in 956 patients. The mean (SD) QTcB was 418 ms (23) and QTcF was 405 ms (20). QTc was found prolonged (>450 ms in women and >440 ms in men) in 129 [13.5%; 95% confidence interval (CI) 11.5, 15.8] and 38 (4%; 95% CI 2.9, 5.4) patients using Bazett and Fridericia corrections, respectively. On multivariable analysis, incomplete bundle branch block, ventricular hypertrophy, signs of ischaemic cardiopathy, female gender, White ethnic origin and age were significantly associated with QTc prolongation. The only HIV variable independently associated with QTc prolongation was the duration of infection (P = 0.023). After adjustment, anti‐HIV treatment, in particular PI (P = 0.99), was not associated with QTc prolongation.

CONCLUSIONS

Although PIs block in vitro hERG current, they are not independently associated with QTc interval prolongation. Prolonged QTc interval in HIV‐infected patients is primarily associated with factors commonly known to prolong QT and with the duration of HIV infection.
The use and potential abuse of anticholinergic antiparkinson drugs in Norway: a pharmacoepidemiological study
Authors: Gjerden Pål , Bramness Jørgen G. , Slørdal Lars
Published: November 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Anticholinergic antiparkinson drugs are used to ameliorate extrapyramidal symptoms caused by either Parkinson's disease or antipsychotic drugs, but their use in the treatment of Parkinson's disease is assumed to be in decline.

• Patients with psychotic conditions have a high prevalence of abuse of drugs, including anticholinergic antiparkinson drugs.

WHAT THIS STUDY ADDS

• Anticholinergic antiparkinson drugs in Norway were primarily prescribed to patients using antipsychotic medication.

• The risk of abuse of this group of drugs was small, even among patients who probably abused other drugs.

AIMS

The use of anticholinergic antiparkinson drugs is assumed to have shifted from the therapy of Parkinson's disease to the amelioration of extrapyramidal adverse effects induced by antipsychotic drugs. There is a considerable body of data suggesting that anticholinergic antiparkinson drugs have a potential for abuse. The aim was to investigate the use and potential abuse of this class of drugs in Norway.

METHODS

Data were drawn from the Norwegian Prescription Database on sales to a total of 73 964 patients in 2004 of biperiden and orphenadrine, and use in patients with Parkinson's disease or in patients who were also prescribed antipsychotic agents. Possible abuse of these drugs was assessed by the level of use, skewedness of use, indications of drug‐seeking behaviour and concomitant use of benzodiazepine tranquillizers, a group of prescription drugs with a recognized potential for abuse.

RESULTS

Anticholinergic antiparkinson drugs were prescribed to 4.5% of all outpatients who used antipsychotic drugs. This outnumbered sales to patients with Parkinson's disease by >20 to 1. We found indications of abuse of benzodiazepine tranquillizers among patients using antipsychotics, but there were no clear indications of abuse of anticholinergics, even among patients who were strongly suspected of abuse of benzodiazepines.

CONCLUSIONS

Anticholinergic antiparkinson drugs were used primarily by patients with psychotic illnesses. These patients have a very high prevalence of legal and illegal drug abuse, but the risk of abuse of anticholinergic antiparkinson drugs seemed small.
The availability of prescription‐only analgesics purchased from the internet in the UK
Authors: Raine Connie , Webb David J. , Maxwell Simon R. J.
Published: November 2008

WHAT IS ALREADY KNOWN ABOUT THIS SUBJECT

• Increasing numbers of people are accessing medicines from the internet.

• This online market is poorly regulated and represents a potential threat to the health of patients and members of the public.

WHAT THIS STUDY ADDS

• Prescription‐only analgesics, including controlled opioids, are readily available to the UK public through internet pharmacies that are easily identified by popular search engines.

• The majority of websites do not require the customer to possess a valid prescription for the drug.

• Less than half provide an online health screen to assess suitability for supply.

• The majority have no registered geographical location.

• Analgesic medicines are usually purchased at prices significantly above British National Formulary prices and are often supplied in large quantities.

• These findings are of particular relevance to pain‐management specialists who are trying to improve the rational use of analgesic drugs.

AIMS To explore the availability to the UK population of prescription‐only analgesics from the internet.

METHODS Websites were identified by using several keywords in the most popular internet search engines. From 2000 websites, details of 96 were entered into a database.

RESULTS Forty‐six (48%) websites sold prescription analgesics, including seven opioids, two non‐opioids and 18 nonsteroidal anti‐inflammatory drugs. Thirty‐five (76%) of these did not require the customer to possess a valid prescription.

CONCLUSION Prescription‐only analgesics, including controlled opioids, are readily available from internet websites, often without a valid prescription.
Synergistic effects of cyclic strain and Th1-like cytokines on tenascin-C production by rheumatic aortic valve interstitial cells
Authors: Jiang L , Wei X F , Yi D H , Xu P , Liu H , Chang Q , Yang S M , Li Z F , Gao H B , Hao G J
Published: November 2008

Tenascin-C (TN-C) is a key component of extracellular matrix (ECM) and its expression process is poorly understood during rheumatic heart valvular disease (RHVD). In this study, we found that interferon (IFN)-γ, tumour necrosis factor (TNF)-α and TN-C concentrations in patients with RHVD were significantly higher than in normal controls. More IFN-γ receptors and TNF receptors were found being expressed on rheumatic aortic valves interstitial cells than on non-rheumatic ones and their expression was patients' sera dependent. Antibodies neutralizing IFN-γ or TNF-α could attenuate patients' sera-induced TN-C transcription by isolated rheumatic aortic valves interstitial cells. By application with different protein kinase inhibitors, we found that combined with cyclic strain, TNF-α and IFN-γ induced TN-C transcription through the RhoA/ROCK signalling pathway. At the same time, p38 mitogen-activated protein kinase was involved in TNF-α and IFN-γ induced TN-C transcription. TNF-α also increased TN-C mRNA level by additional PKC and ERK 1/2 activation. Our finding revealed a new insight into ECM remodelling during RHVD pathogenesis and new mechanisms involved in the clinical anti-IFN-γ and anti-TNF-α therapy.
The hinge region fragment of immunoglobulin G improves immunogenicity of recombinant gonadotrophin‐releasing hormone conjugated to the T‐helper epitope in designing peptide vaccines
Authors: Xu Jinshu , Wu Jie , Wang Xuejun , Zhang Yin , Li Wenjia , Zhu Zheng , Zhu Dongya , Hu Zhuoyi , Roque Rouel S. , Liu Jingjing
Published: November 2008

In our previous study, the hinge fragment (225–232/225′–232′) of human immunoglobulin G1 (IgG1) was used as a space peptide linker for synthesizing the GnRH3–hinge–MVP chimeric peptide, whereby three repeated gonadotrophin‐releasing hormone (GnRH) units and a T‐cell epitope from measles virus fusion protein (MVP) were amide‐bond‐linked at the N and C terminus, respectively, to the hinge peptide for producing anti‐GnRH antibody responses. To investigate whether or not the hinge region fragment can improve the immunogenicity of GnRH, we further synthesized and purified GnRH3–hinge–MVP, GnRH3–hinge and GnRH3–MVP using recombinant DNA technology. Under high pH conditions, GnRH3–hinge–MVP was capable of forming double‐chain structures. Immunization of male mice with the immunogens of GnRH3–hinge–MVP resulted in the generation of high‐titre antibodies specific for GnRH. The synthetic GnRH3–hinge and GnRH3–MVP induced a lower titre of anti‐GnRH antibody than GnRH3–hinge–MVP. This was followed by a decrease in serum testosterone levels, which resulted in a low level of expression of the relaxin‐like factor gene in the testis. Our data suggest that peptide and T‐cell epitopes oriented at the N‐terminus or C‐terminus of hinge peptides simplify the antigenic peptide conjugates and may be considered as potential synthetic immunogens.
Identification of the proteome composition occurring during the course of embryonic development of bees (Apis mellifera)
Authors: Li J. , Zhang L. , Feng M. , Zhang Z. , Pan Y.
Published: November 2008

To investigate the proteome during embryonic development of honeybees, Apis mellifera, proteins were identified by two‐dimensional gel electrophoresis, mass spectrometry and protein engine identification tools that were applied to MASCOT and Xproteo search engines. 312, 320, 315 proteins were detected in 24, 48 and 72 h embryos. Thirty‐eight highly abundant proteins were identified at the three time points by MS fingerprinting. All 21 proteins could be identified as products of annotated genes of the honeybee. Identified proteins included six proteins related to the metabolism of carbohydrates and energy production, six proteins belonging to the heat shock protein family, three cytoskeletal proteins, four proteins related to the antioxidant system of the embryo and two proteins related to growth regulation of the embryo. Quantitative proteomics was applied to analyze differences in amounts of these proteins during the three above mentioned developmental stages. Our data present an initial molecular picture of honeybee embryos, and will hopefully pave the way for future research on this animal.
Habitat structure mediates predation risk for sedentary prey: experimental tests of alternative hypotheses
Authors: Chalfoun Anna D. , Martin Thomas E.
Published: November 2008

Predation is an important and ubiquitous selective force that can shape habitat preferences of prey species, but tests of alternative mechanistic hypotheses of habitat influences on predation risk are lacking.

We studied predation risk at nest sites of a passerine bird and tested two hypotheses based on theories of predator foraging behaviour. The total‐foliage hypothesis predicts that predation will decline in areas of greater overall vegetation density by impeding cues for detection by predators. The potential‐prey‐site hypothesis predicts that predation decreases where predators must search more unoccupied potential nest sites.

Both observational data and results from a habitat manipulation provided clear support for the potential‐prey‐site hypothesis and rejection of the total‐foliage hypothesis. Birds chose nest patches containing both greater total foliage and potential nest site density (which were correlated in their abundance) than at random sites, yet only potential nest site density significantly influenced nest predation risk.

Our results therefore provided a clear and rare example of adaptive nest site selection that would have been missed had structural complexity or total vegetation density been considered alone.

Our results also demonstrated that interactions between predator foraging success and habitat structure can be more complex than simple impedance or occlusion by vegetation.
Effect of current reproduction on apparent survival, breeding dispersal, and future reproduction in barn swallows assessed by multistate capture–recapture models
Authors: Schaub Michael , Von Hirschheydt Johann
Published: November 2008

Theoretical models predict a negative effect of current reproduction on breeding dispersal, survival and future reproduction, and many studies confirm these predictions. Yet, results of most previous studies may be difficult to interpret because the fate of the affected individuals cannot always be observed. Detection is almost always imperfect and some individuals emigrate from the study area, resulting in biased estimates of both survival and dispersal.

Most studies bypass these problems with strong assumptions. We use a multistate capture–recapture model that does not require these assumptions. States are defined based on classes of reproductive success and on observed dispersal events within the study area. By accounting for imperfect detection within the study area, the model allows estimation of the effect of reproductive success on apparent survival, dispersal probabilities within the study area and the annual transition probabilities among classes of reproductive success. Based on an assumption about the estimate of real survival, the model allows the estimation of total dispersal that is not specific to a fixed study area.

We applied this model to capture–recapture data of 2262 adult barn swallows (Hirundo rustica) sampled from 1997–2004 in eight local populations in Switzerland.

We found that dispersal within the study area decreased with increasing reproductive success in both sexes, that reproductive success was not affected by preceding dispersal and that apparent survival of females but not of males increased with increasing reproductive success. Apparent survival of females with high reproductive success was identical to apparent survival of males suggesting that this estimate of apparent survival (0·48) was close to true survival. Total breeding dispersal was generally higher in females and it increased with decreasing reproductive success in both sexes. Current reproductive success depended on reproductive success in the preceding year suggesting that individual differences were of importance.

Our study highlights that reproductive success was an important factor affecting breeding dispersal and population turnover. While unsuccessful males mainly remained in the local populations, many unsuccessful females left them. Population turnover of adult swallows was mainly due to unsuccessful females.
The long‐term ecology of the lost forests of La Laguna, Tenerife (Canary Islands)
Authors: De Nascimento Lea , Willis Katherine J. , Fernández‐Palacios José María , Criado Constantino , Whittaker Robert J.
Published: November 2008

Aim  We report the first analysis of the long‐term ecology of Tenerife, in order to establish a pre‐colonization base‐line and to assess the effect of human activity and the role of climatic variation on vegetation during the Late Holocene.

Location  A former lake bed in the city of La Laguna (Tenerife, Canary Islands, Spain).

Methods  A sedimentary sequence of over 2 m was obtained from the former lake bed. Fossil pollen and microfossil charcoal concentrations were analysed. Radiocarbon dating of the sequence indicates that it spans approximately the last 4700 years. The pollen diagram was zoned using optimal splitting within psimpoll 4.25.

Results  Three pollen zones were differentiated: (1) in Zone L1 (c. 4700–2900 cal. yr bp) a mixed forest was dominated by Quercus, Carpinus, Myrica and Pinus; (2) in Zone L2 (c. 2900–2000 cal. yr bp) the laurel forest taxa increased, while Pinus, Juniperus and Phoenix declined; and (3) Zone L3 (c. 2000–400 cal. yr bp) was characterized by the decline of Carpinus and Quercus and the abundance of laurel forest taxa (e.g. Myrica). Neither Carpinus nor Quercus was hitherto considered to be native to the Canary Islands. Their decline started c. 2000 years ago, coinciding with microfossil charcoal evidence of increased burning and with archaeological evidence for the first human settlement on Tenerife.

Main conclusions  Between c. 4700 and 2000 cal. yr bp, the composition of the forest in the valley of La Laguna was very different from what it is at present. In particular, Quercus and Carpinus appear to have been significant components, alongside components of the present‐day laurel forest, and the native pine (Pinus canariensis) forest and thermophilous woodland were also more prevalent in the region (but probably not within the lake basin itself) until 3000 cal. yr bp. The subsequent decline of Quercus and Carpinus led to the establishment of the present laurel forest in the region and a shift to more open vegetation types. These changes indicate that the aboriginal inhabitants of the islands, the Guanches, had a far more profound impact on the vegetation of Tenerife than hitherto realized.
A new methodology for reconstructing climate and vegetation from modern pollen assemblages: an example from British Columbia
Authors: Goring Simon , Pellatt Marlow G. , Lacourse Terri , Walker Ian R. , Mathewes Rolf W.
Published: November 2008

Aim We used modern pollen assemblages to develop a method for climate reconstruction that reduces the spatial autocorrelation of residuals and accounts for the strong topographic and climatic variation that occurs in British Columbia, Canada.

Location British Columbia, Canada, including sites both on the mainland and on adjacent islands (Queen Charlotte Islands and Vancouver Island).

Methods New pollen assemblages from surface‐sediment samples collected in British Columbia were combined with other published and unpublished samples (n = 284). Multivariate rank‐distances between sample sites and a randomized set of sites within the province were calculated for climate parameters to determine whether gaps in the current network of present‐day pollen sample sites exist. Lacustrine surface‐sediment pollen assemblages (n = 145) were ordinated using non‐metric multidimensional scaling (NMDS), and a generalized additive model (GAM) was used to reconstruct modern mean warmest month temperature (MWMT) and mean annual precipitation (MAP) from the NMDS ordination. The results were compared with standard climate reconstruction techniques, including the modern analogue technique, partial least squares, weighted averaging, weighted averaging–partial least squares and factor analysis.

Results Reconstructions of MWMT and MAP using NMDS and GAM were comparable to those of existing models. When reconstructing MWMT, the NMDS/GAM method had a lower root‐mean‐squared error of prediction (RMSEP), lower spatial autocorrelation and higher correlation with observed temperature values than the other methods tested. When reconstructing MAP, the partial least squares method performed better than the NMDS/GAM method for RMSEP and correlation with observed values; however, the NMDS/GAM method had a lower spatial autocorrelation of residuals.

Main conclusions NMDS reveals strong relationships among modern pollen assemblages, vegetation and climate parameters. Climate models using NMDS and GAM are comparable to other palaeoecological reconstruction models, but provide lower spatial autocorrelation of residuals for both parameters tested. An inverse distance‐weighted surface of multivariate rank‐climate distances generated from the network of pollen sample sites indicates that greater sampling intensity in north‐western and central‐interior British Columbia is required in order to obtain an accurate representation of climatic and vegetation diversity in the province.
The role of dispersal ability, climate and spatial separation in shaping biogeographical patterns of phylogenetically distant plant groups in seasonally dry Andean forests of Bolivia
Authors: Linares‐Palomino Reynaldo , Kessler Michael
Published: November 2008

Aim  To assess biogeographical patterns of Acanthaceae, Bromeliaceae, Cactaceae and Pteridophyta in Bolivian Andean seasonally dry forest islands and to explain current floristic differences between these islands by means of extrinsic (precipitation, elevation) and intrinsic (dispersal ability) factors.

Location  Ten isolated and disjunct seasonally dry forest areas in the Bolivian Andes and the adjacent seasonal forest areas of the Chiquitanía and Chaco regions.

Methods  We collated species data from recently updated and revised taxonomic treatments and herbarium collections for Acanthaceae, Bromeliaceae, Cactaceae and Pteridophyta, constructed floristic distance matrices to estimate beta diversity at the study sites and subjected them to Mantel correlation analyses. Multiple regressions on distance matrices allowed us to test the influence of geographical distance and environmental (elevation and precipitation) differences on floristic differentiation.

Results  Acanthaceae and Bromeliaceae, and to a lesser extent Cactaceae, showed coincident biogeographical patterns and suggested the presence of two seasonally dry forest groups in Andean Bolivia: one including all small isolated northern dry valleys and another including all southern valleys with connections to the lowland seasonal forests of the Chiquitanía and the Chaco. Most of the variation in the floristic distance matrices in these plant groups, with seed dispersal typically restricted to short distances, was explained by spatial separation between habitat islands. In contrast, pteridophytes showed a different biogeographical pattern. Their floristic differences between sites were determined by the environmental variables. The anemochorous and spore‐based dispersal system of this plant group seems to be a highly effective mechanism allowing pteridophytes to easily reach even the isolated dry valleys in inter‐Andean Bolivia.

Main conclusions  Current biogeographical patterns in dry Andean habitat islands can provide insights into the factors that control the processes of community assembly. We show that differences in community composition of phylogenetically distant plant groups in the understorey of seasonally dry forest islands can be explained by a combination of the habitat characteristics where the group is present (either precipitation, elevation or both) and, more interestingly, by group specific dispersal limitation (as inferred by geographical distance between island habitats).
Free fatty acid kinetics during long‐term treatment with pioglitazone added to sulfonylurea or metformin in Type 2 diabetes
Authors: Roden M. , Mariz S. , Brazzale A. R. , Pacini G.
Published: November 2008

Background.  Free fatty acids (FFAs) are linked to impaired insulin action, but their role in mediating long‐term insulin sensitization during diabetes treatment is unclear.

Objectives.  To examine the effect of pioglitazone addition to existing therapy on FFA dynamics and insulin action.

Design.  Two 2‐year, randomized, parallel‐group, double‐blind, double‐dummy, clinical trials.

Setting.  One hundred and seventy‐one centres in Europe, Australia and Canada.

Subjects.  Male and female patients with Type 2 diabetes inadequately managed with metformin or sulfonylurea.

Interventions.  Patients were randomized to pioglitazone (15–45 mg day−1; n = 319) or metformin (850–2550 mg day−1; n = 320) as add‐on therapy to gliclazide or pioglitazone (n = 317) versus gliclazide (80–320 mg day−1; n = 313) as add‐on therapy to metformin.

Outcome measure.  Plasma FFA profiles during oral glucose tolerance tests in selected centres before and during treatment (n = 588).

Results.  At Week 104, pioglitazone treatment decreased fasting FFAs by 0.08 mmol L−1 when added to sulfonylurea and by 0.11 mmol L−1 when added to metformin versus the respective sulfonylurea + metformin groups (0.03 mmol L−1, P = 0.05 and 0.04 mmol L−1, P < 0.05), and this was accompanied by significant improvements in fasting adipose tissue insulin sensitivity. Changes in postchallenge FFAs were similar between groups and not related to changes in liver transaminases, insulin action and secretion. However, the sensitivity of FFA to insulin was affected by treatment (P < 0.001) and visit (P < 0.05). Insulin sensitivity of FFA rose when pioglitazone was added to sulfonylurea (P < 0.05), but decreased for gliclazide + metformin (P < 0.05).

Conclusion.  Long‐term improvements in adipose tissue insulin sensitivity and reduction in fasting FFAs with pioglitazone may help to reduce lipotoxicity in Type 2 diabetes.
Effect of prolonged erosive pH cycling on different restorative materials
Authors: HONÓRIO H. M. , RIOS D. , FRANCISCONI L. F. , MAGALHÃES A. C. , MACHADO M. A. A. M. , BUZALAF M. A. R.
Published: November 2008

Summary  This in vitro study evaluated the effect of a prolonged erosive pH cycling on the superficial microhardness change (SMHC) and the erosive wear of different restorative materials. Eighty enamel specimens with prepared cavities of 1·5 × 1·5 mm were randomly divided into eight groups according to the restorative materials used for the fillings (RMGI – resin‐modified glass–ionomer, CGI – conventional glass–ionomer, CR– composite resin, A – amalgam) and immersion media used (ERO – erosive medium or SAL – artificial saliva). During 35 days, half of the specimens were immersed in a cola drink (ERO), for 5 min, three times a day, and they remained in SAL between the erosive cycles. The other half of the specimens was immersed in SAL only, for the entire experimental period (control). Data were tested for significant differences by anova and Tukey’s tests (P < 0·05). Scanning electron microscopy images were made to illustrate the enamel erosive wear and restorative materials alterations. The mean SMHC (%) and mean erosive wear (μm) of the materials were: RMGI‐ERO (30/0·5); CGI‐ERO (37/0·5); CR‐ERO (−0·3/0·3); A‐ERO (−4/0·3); RMGI‐SAL (4/0·4); CGI‐SAL (−6/0·4); CR‐SAL (−3/0·2) and A‐SAL (2/0·4). Scanning electron microscopy images showed pronounced enamel erosive wear on groups submitted to erosive pH cycling when compared with groups maintained in saliva. In conclusion, the prolonged pH cycling promoted significantly higher alterations (SMHC and erosive wear) on the glass–ionomer cements than the CR and amalgam.
Morphological and molecular changes in denture‐supporting tissues under persistent mechanical stress in rats
Authors: TSURUOKA M. , ISHIZAKI K. , SAKURAI K. , MATSUZAKA K. , INOUE T.
Published: November 2008

Summary  The purpose of this study was to determine the effects of mechanical compression on the palatal mucosa using an experimental palatal base. The palatal base was either pressed onto (stress group) or not pressed onto (fit group) rat palatal mucosa. Blood flow was measured and the animals were sacrificed 6–72 h later for analysis. The expression of heat shock protein 70 (HSP70), vascular endothelial growth factor (VEGF) and proliferation cell nuclear antigen (PCNA) was characterized by immunohistochemical staining. For morphometric analysis, connective tissues were divided into bone side and epithelial side tissues. The ratio of PCNA‐positive cells (PCNA score) was calculated, and the expressions of mRNA encoding HSP70 and VEGF was evaluated. Whereas blood flow in the stress group showed ischaemia, none was found in the fit group. Proliferation cell nuclear antigen scores on the bone side were higher than on the epithelial side in the stress group (P < 0·05). Heat shock protein 70‐ and VEGF‐positive cells were observed under compression conditions, particularly in the periosteum. In the stress group, the expressions of mRNA encoding HSP70 and VEGF were highest at 12 h (P < 0·05). These results suggest that mechanical compression of the palatal plate induces ischaemia, and that cells in the underlying denture‐supporting tissue, which includes the periosteum, synthesize HSP70 and VEGF to maintain homeostasis under these conditions.
Investigation of infraorbital nerve injury following zygomaticomaxillary complex fractures
Authors: SAKAVICIUS D. , JUODZBALYS G. , KUBILIUS R. , SABALYS G. P.
Published: November 2008

Summary  The aim of this study was to investigate the severity of infraorbital nerve injury following zygomaticomaxillary complex fractures and to estimate the treatment methods facilitating its functional recovery. A total of 478 patients with unilateral zygomaticomaxillary complex fractures were treated. Infraorbital nerve sensory disturbances were diagnosed in 64·4% of the patients. Injury of the infraorbital nerve was expressed as asymmetry index, which was calculated as a ratio between the affected side and the intact side electric pain detection thresholds at the innervation zone skin before treatment and 14 days, 1, 3, 6 and 12 months postoperatively. A mean asymmetry index of 0·6 ± 0·03 and 1·9 ± 0·5 was registered for 57 (11·9%) patients with hyperalgesia and for 251 (52·5%) patients with hypoalgesia, respectively. As a result of retrospective analysis of infraorbital nerve sensory disturbances and its functional recovery, infraorbital nerve injury severity was classified as mild, moderate and severe. It was found that the dynamics and outcome of the functional infraorbital nerve recovery depend on the severity of the injury and the presence of infraorbital canal damage. Function was completely recovered within 3 months after treatment in cases with mild nerve injury. In moderate cases, complete recovery was seen within 6 months and in 34·6% of the severe cases, within a 12‐month period after treatment when infraorbital nerve decompression was performed according to the stated indication. Treatment based on infraorbital nerve injury classification offers a better prognosis for complete recovery of the infraorbital nerve function.
Low‐level laser therapy improves bone repair in rats treated with anti‐inflammatory drugs
Authors: RIBEIRO D. A. , MATSUMOTO M. A.
Published: November 2008

Summary  Nowadays, selective cyclooxygenase‐2 non‐steroidal anti‐inflammatory drugs have been largely used in surgical practice for reducing oedema and pain. However, the association between these drugs and laser therapy is not known up to now. Herein, the aim of this study was to evaluate the action of anti‐COX‐2 selective drug (celecoxib) on bone repair associated with laser therapy. A total of 64 rats underwent surgical bone defects in their tibias, being randomly distributed into four groups: Group 1) negative control; Group 2) animals treated with celecoxib; Group 3) animals treated with low‐level power laser and Group 4) animals treated with celecoxib and low‐level power laser. The animals were killed after 48 h, 7, 14 and 21 days. The tibias were removed for morphological, morphometric and immunohistochemistry analysis for COX‐2. Statistical significant differences (P < 0·05) were observed in the quality of bone repair and quantity of formed bone between groups at 14 days after surgery for Groups 3 and 4. COX‐2 immunoreactivity was more intense in bone cells for intermediate periods evaluated in the laser‐exposed groups. Taken together, such results suggest that low‐level laser therapy is able to improve bone repair in the tibia of rats as a result of an up‐regulation for cyclooxygenase‐2 expression in bone cells.
Effect of turban use on temporomandibular joint problems
Authors: UYSAL S. , AVCU N.
Published: November 2008

Summary  The aim of this study was to evaluate the effects of daily turban wear on temporomandibular joint (TMJ) problems. This cross‐sectional study was carried out on 249 female patients. Of these, 119 patients were using turban daily, while 130 patients did not use. Patients were asked questions to ascertain the signs of TMJ problems. Thereafter, the TMJ region was examined clinically. Cross‐tabulations and Chi‐square statistics were computed in accordance with Bonferroni correction for multiple comparisons. To investigate the association between continuous turban wear and temporomandibular disorder symptoms, logistic regression analysis was performed. Limited mouth opening, deviation, pain on TMJ palpation and mouth opening were not affected with turban usage. However, turban users more frequently demonstrated pain during palpation of the masticatory muscles than non‐users (P = 0·001). Duration of the turban usage did not affect clinical examination findings except pain on masticatory muscles during palpation (P = 0·001). Complaint of pain on masticatory muscles are more frequently seen among the turban users.
Characterization of the dTDP‐d‐fucofuranose biosynthetic pathway in Escherichia coli O52
Authors: Wang Quan , Ding Peng , Perepelov Andrei V. , Xu Yanli , Wang Ying , Knirel Yuriy A. , Wang Lei , Feng Lu
Published: November 2008

d‐fucofuranose (d‐Fucf) is a component of Escherichia coli O52 O antigen. This uncommon sugar is also the sugar moiety of the anticancer drug – gilvocarcin V produced by many streptomycetes. In E. coli O52, rmlA, rmlB, fcf1 and fcf2 were proposed in a previous study by our group to encode the enzymes of the dTDP‐d‐Fucf (the nucleotide‐activated form of d‐Fucf) biosynthetic pathway. In this study, Fcf1 and Fcf2 from E. coli O52 were expressed, purified and assayed for their respective activities. Novel product peaks from enzyme‐substrate reactions were detected by capillary electrophoresis and the structures of the product compounds were elucidated by electro‐spray ionization mass spectrometry and nuclear magnetic resonance spectroscopy. Fcf1 was confirmed to be a dTDP‐6‐deoxy‐d‐xylo‐hex‐4‐ulopyranose reductase for the conversion of dTDP‐6‐deoxy‐d‐xylo‐hex‐4‐ulopyranose to dTDP‐d‐fucopyranose (dTDP‐d‐Fucp), and Fcf2 a dTDP‐d‐Fucp mutase for the conversion of dTDP‐d‐Fucp to dTDP‐d‐Fucf. The Km of Fcf1 for dTDP‐6‐deoxy‐d‐xylo‐hex‐4‐ulopyranose was determined to be 0.38 mM, and of Fcf2 for dTDP‐d‐Fucp to be 3.43 mM. The functional role of fcf1 and fcf2 in the biosynthesis of E. coli O52 O antigen were confirmed by mutation and complementation tests. This is the first time that the biosynthetic pathway of dTDP‐d‐Fucf has been fully characterized.
Regulation of glutamate metabolism by protein kinases in mycobacteria
Authors: O'Hare Helen M. , Durán Rosario , Cerveñansky Carlos , Bellinzoni Marco , Wehenkel Anne Marie , Pritsch Otto , Obal Gonzalo , Baumgartner Jens , Vialaret Jérome , Johnsson Kai , Alzari Pedro M.
Published: November 2008

Protein kinase G of Mycobacterium tuberculosis has been implicated in virulence and in regulation of glutamate metabolism. Here we show that this kinase undergoes a pattern of autophosphorylation that is distinct from that of other M. tuberculosis protein kinases characterized to date and we identify GarA as a substrate for phosphorylation by PknG. Autophosphorylation of PknG has little effect on kinase activity but promotes binding to GarA, an interaction that is also detected in living mycobacteria. PknG phosphorylates GarA at threonine 21, adjacent to the residue phosphorylated by PknB (T22), and these two phosphorylation events are mutually exclusive. Like the homologue OdhI from Corynebacterium glutamicum, the unphosphorylated form of GarA is shown to inhibit α‐ketoglutarate decarboxylase in the TCA cycle. Additionally GarA is found to bind and modulate the activity of a large NAD+‐specific glutamate dehydrogenase with an unusually low affinity for glutamate. Previous reports of a defect in glutamate metabolism caused by pknG deletion may thus be explained by the effect of unphosphorylated GarA on these two enzyme activities, which may also contribute to the attenuation of virulence.
A single aromatic residue in transcriptional repressor protein KorA is critical for cooperativity with its co‐regulator KorB
Authors: Bingle Lewis E. H. , Rajasekar Karthik V. , Muntaha Sidra tul , Nadella Vinod , Hyde Eva I. , Thomas Christopher M.
Published: November 2008

A central feature of broad host range IncP‐1 plasmids is the set of regulatory circuits that tightly control plasmid core functions under steady‐state conditions. Cooperativity between KorB and either KorA or TrbA repressor proteins is a key element of these circuits and deletion analysis has implicated the conserved C‐terminal domain of KorA and TrbA in this interaction. By NMR we show that KorA and KorB interact directly and identify KorA amino acids that are affected on KorB binding. Studies on mutants showed that tyrosine 84 (or phenylalanine, in some alleles) is dispensable for repressor activity but critical for the specific interaction with KorB in both in vivo reporter gene assays and in vitro electrophoretic mobility shift and co‐purification assays. This confirms that direct and specific protein–protein interactions are responsible for the cooperativity observed between KorB and its corepressors and lays the basis for determining the biological importance of this cooperativity.
The replication fork trap and termination of chromosome replication
Authors: Duggin Iain G. , Wake R. Gerry , Bell Stephen D. , Hill Thomas M.
Published: November 2008

Bacteria that have a circular chromosome with a bidirectional DNA replication origin are thought to utilize a ‘replication fork trap’ to control termination of replication. The fork trap is an arrangement of replication pause sites that ensures that the two replication forks fuse within the terminus region of the chromosome, approximately opposite the origin on the circular map. However, the biological significance of the replication fork trap has been mysterious, as its inactivation has no obvious consequence. Here we review the research that led to the replication fork trap theory, and we aim to integrate several recent findings that contribute towards an understanding of the physiological roles of the replication fork trap. Likely roles include the prevention of over‐replication, and the optimization of post‐replicative mechanisms of chromosome segregation, such as that involving FtsK in Escherichia coli.
A novel component of the division‐site selection system of Bacillus subtilis and a new mode of action for the division inhibitor MinCD
Authors: Bramkamp Marc , Emmins Robyn , Weston Louise , Donovan Catriona , Daniel Richard A. , Errington Jeff
Published: November 2008

Cell division in bacteria is governed by a complex cytokinetic machinery in which the key player is a tubulin homologue, FtsZ. Most rod‐shaped bacteria divide precisely at mid‐cell between segregated sister chromosomes. Selection of the correct site for cell division is thought to be determined by two negative regulatory systems: the nucleoid occlusion system, which prevents division in the vicinity of the chromosomes, and the Min system, which prevents inappropriate division at the cell poles. In Bacillus subtilis recruitment of the division inhibitor MinCD to cell poles depends on DivIVA, and these proteins were thought to be sufficient for Min function. We have now identified a novel component of the division‐site selection system, MinJ, which bridges DivIVA and MinD. minJ mutants are impaired in division because MinCD activity is no longer restricted to cell poles. Although MinCD was thought to act specifically on FtsZ assembly, analysis of minJ and divIVA mutants showed that their block in division occurs downstream of FtsZ. The results support a model in which the main function of the Min system lies in allowing only a single round of division per cell cycle, and that MinCD acts at multiple levels to prevent inappropriate division.
Mhp493 (P216) is a proteolytically processed, cilium and heparin binding protein ofMycoplasma hyopneumoniae
Authors: Wilton Jody , Jenkins Cheryl , Cordwell Stuart J. , Falconer Linda , Minion F. Chris , Oneal David C. , Djordjevic Michael A. , Connolly Angela , Barchia Idris , Walker Mark J. , Djordjevic Steven P.
Published: November 2008

Mycoplasma hyopneumoniaeinduces respiratory disease in swine by colonizing cilia causing ciliostasis, cilial loss and epithelial cell death. Heparin binds toM. hyopneumoniaecells in a dose‐dependent manner and blocks its ability to adhere to porcine cilia. We show here that Mhp493 (P216), a paralogue of the cilium adhesin P97 (Mhp183), is cleaved between amino acids 1040 and 1089 generating surface‐accessible, heparin‐binding proteins P120 and P85. Antiphosphoserine antibodies recognized P85 in 2‐D immunoblotting studies and TiO2chromatography of trypsin digests of P85 isolated a single peptide with an m/z of 917.3. A phosphoserine residue in the tryptic peptide90VSELpSFR96(position 94 in P85) was identified by MALDI‐MS/MS. Polyhistidine fusion proteins (F1P216, F2P216, F3P216) spanning Mhp493 bound heparin with biologically significant Kd values, and heparin, fucoidan and mucin inhibited this interaction. Latex beads coated with F1P216, F2P216and F3P216adhered to and entered porcine kidney epithelial‐like (PK15) cell monolayers. Microtitre plate‐based assays showed that sequences within P120 and P85 bind to porcine cilia and are recognized by serum antibodies elicited during infection byM. hyopneumoniae. Mhp493 contributes significantly to the surface architecture ofM. hyopneumoniaeand is the first cilium adhesin to be described that lacks an R1 cilium‐binding domain.
Mycobacterium tuberculosis expresses methionine sulphoxide reductases A and B that protect from killing by nitrite and hypochlorite
Authors: Lee Warren L. , Gold Benjamin , Darby Crystal , Brot Nathan , Jiang Xiuju , De Carvalho Luiz Pedro S. , Wellner Daniel , St. John Gregory , Jacobs Jr William R. , Nathan Carl
Published: November 2008

Methionine sulphoxide reductases (Msr) reduce methionine sulphoxide to methionine and protect bacteria against reactive oxygen intermediates (ROI) and reactive nitrogen intermediates (RNI). Many organisms express both MsrA, active against methionine‐(S)‐sulphoxide, and MsrB, active against methionine‐(R)‐sulphoxide. Mycobacterium tuberculosis (Mtb) expresses MsrA, which protects ΔmsrA‐Escherichia coli from ROI and RNI. However, the function of MsrA in Mtb has not been defined, and it is unknown whether Mtb expresses MsrB. We identified MsrB as the protein encoded by Rv2674 in Mtb and confirmed the distinct stereospecificities of recombinant Mtb MsrA and MsrB. We generated strains of Mtb deficient in MsrA, MsrB or both and complemented the mutants. Lysates of singly deficient strains displayed half as much Msr activity as wild type against N‐acetyl methionine sulphoxide. However, in contrast to other bacteria, single mutants were no more vulnerable than wild type to killing by ROI/RNI. Only Mtb lacking both MsrA and MsrB was more readily killed by nitrite or hypochlorite. Thus, MsrA and MsrB contribute to the enzymatic defences of Mtb against ROI and RNI.