bluuebunny/crossref_metadata_2025 huggingface dataset:
In vitro reaction of human osteoblasts on alumina‐toughened zirconia
Authors: Kohal Ralf J. , Baechle Maria , Han Jung S. , Hueren Dennis , Huebner Ute , Butz Frank
Published: August 2009

Objectives: Alumina toughening enhances the mechanical properties of zirconia ceramics but the biocompatibility of this material has rarely been addressed. In this study, we examined the osteoblast response to alumina‐toughened zirconia (ATZ) with different surface topographies.

Material and methods: Human osteoblasts isolated from maxillary biopsies of four patients were cultured and seeded onto disks of the following substrates: ATZ with a machined surface, airborne‐particle abraded ATZ, airborne‐particle abraded and acid etched ATZ. Airborne‐particle abraded and acid etched titanium (SLA) and polystyrene disks served as a reference control. The surface topography of the various substrates was characterized by profilometry (Ra, Rp−v) and scanning electron microscopy (SEM). Cell proliferation, cell‐covered surface area, alkaline phophatase (ALP) and osteocalcin production were determined. The cell morphology was analyzed on SEM images.

Results: The surface roughness of ATZ was increased by airborne‐particle abrasion, but with the Ra and Rp−v values showing significantly lower values compared with SLA titanium (Mann‐Whitney U‐test P<0.05). The proliferation assay revealed no statistically significant differences between the ATZ substrates, SLA titanium and polystyrene (Kruskal–Wallis test, P>0.05). All substrates were densely covered by osteoblasts. ALP and osteocalcin production was similar on the examined surfaces. Cell morphology analysis revealed flat‐spread osteoblasts with cellular extensions on all substrates.

Conclusions: These results indicate that ATZ may be a viable substrate for the growth and differentiation of human osteoblasts. Surface modification of ATZ by airborne‐particle abrasion alone or in combination with acid etching seems not to interfere with the growth and differentiation of the osteoblasts.
Melatonin in diseases of the oral cavity
Authors: Gómez‐Moreno G , Guardia J , Ferrera MJ , Cutando A , Reiter RJ
Published: August 2009

Oral Diseases (2010) 16, 242–247

Background:  Melatonin is the principal secretory product of the pineal gland. It has immunomodulatory and antioxidant activities, stimulates the proliferation of collagen and osseous tissue and acts as a protector against cellular degeneration associated with aging and toxin exposure. Arising out of its antioxidant actions, melatonin protects against inflammatory processes and cellular damage caused by the toxic derivates of oxygen. As a result of these actions, melatonin may be useful as a co‐adjuvant in the treatment of certain conditions of the oral cavity.

Methods:  An extensive review of the scientific literature was carried out using PubMed, Science Direct, ISI Web of Knowledge and the Cochrane base.

Results:  Melatonin, which is released into the saliva, may have important implications for oral diseases. Melatonin may have beneficial effects in certain oral pathologies including periodontal diseases, herpes viral infections and Candida, local inflammatory rocesses, xerostomia, oral ulcers and oral cancer.

Conclusions:  Melatonin may play a role in protecting the oral cavity from tissue damage caused by oxidative stress. The experimental evidence suggests that melatonin may have utility in the treatment of several common diseases of the oral cavity. However, more specific studies are necessary to extend the therapeutic possibilities to other oral diseases.
Implant‐Supported Electrostimulating Device to Treat Xerostomia: A Preliminary Study
Authors: Ami Smidt , Wolff Andy
Published: August 2009

Background: The full accomplishment of salivary function depends on proper salivary flow rate and composition. Salivary secretion is highly essential in the maintenance of health and integrity of oral hard and soft tissue. Xerostomia is a common symptom affecting between one‐fifth and one‐third of the adult population, more commonly women than men. Induction of salivary secretion exists in several pharmacological formulations per os. Electrostimulation to enhance salivary secretion has been used frequently as a research tool but only in limited extent as a clinical method to treat patients with xerostomia.

Purpose: The aims of this preliminary study were to observe and evaluate the therapeutic effect on xerostomia of the Saliwell Crown (Saliwell Ltd., Harutzim, Israel), an innovative saliva electrostimulation device fixed on an implant, placed in the lower third molar area.

Materials and Methods: A Saliwell Crown was placed in the lower third molar area of an 81‐year‐old female patient with complaints of dry and burning mouth. Salivary secretion was measured, and the patient was asked to fill in written satisfaction questionnaires. The patient was monitored for a year, comparing her salivary secretion rates and the written questionnaires.

Results: The results showed a constant slight but significant increase in the salivary secretion and in the patient's personal feelings as presented in the questionnaires.

Conclusions: The saliva stimulation device Saliwell Crown, placed on an implant in an 81‐year‐old patient with dry and burning mouth complaints, presented promising results when both the salivary secretion tests and the self‐assessment questionnaires were examined and compared.
A mutagenic analysis of the RNase mechanism of the bacterial Kid toxin by mass spectrometry
Authors: Diago‐Navarro Elizabeth , Kamphuis Monique B. , Boelens Rolf , Barendregt Arjan , Heck Albert J. , van den Heuvel Robert H. , Díaz‐Orejas Ramón
Published: August 2009

Kid, the toxin of the parD (kis, kid) maintenance system of plasmid R1, is an endoribonuclease that preferentially cleaves RNA at the 5′ of A in the core sequence 5′‐UA(A/C)‐3′. A model of the Kid toxin interacting with the uncleavable mimetic 5′‐AdUACA‐3′ is available. To evaluate this model, a significant collection of mutants in some of the key residues proposed to be involved in RNA binding (T46, A55, T69 and R85) or RNA cleavage (R73, D75 and H17) were analysed by mass spectrometry in RNA binding and cleavage assays. A pair of substrates, 5′‐AUACA‐3′, and its uncleavable mimetic 5′‐AdUACA‐3′, used to establish the model and structure of the Kid–RNA complex, were used in both the RNA cleavage and binding assays. A second RNA substrate, 5′‐UUACU‐3′ efficiently cleaved by Kid both in vivo and in vitro, was also used in the cleavage assays. Compared with the wild‐type protein, mutations in the residues of the catalytic site abolished RNA cleavage without substantially altering RNA binding. Mutations in residues proposed to be involved in RNA binding show reduced binding efficiency and a corresponding decrease in RNA cleavage efficiency. The cleavage profiles of the different mutants were similar with the two substrates used, but RNA cleavage required much lower protein concentrations when the 5′‐UUACU‐3′ substrate was used. Protein synthesis and growth assays are consistent with there being a correlation between the RNase activity of Kid and its inhibitory potential. These results give important support to the available models of Kid RNase and the Kid–RNA complex.
Evaluating the Role of the Serotoninergic System in the Control of Human Seminal Vesicle Smooth Muscle—An in Vitro Approach
Authors: Birowo Ponco , Ückert Stefan , Kedia George T. , Scheller Friedemann , Meyer Markus , Taher Akmal , Rahardjo Djoko , Jonas Udo , Kuczyk Markus A.
Published: August 2009

Introduction

It has been suggested that serotonin re-uptake inhibitors (SRIs) may retard the ejaculatory response by acting directly on the seminal vesicle (SV) and ductus deferens smooth muscle. However, until now, only a very few experimental studies have investigated such potential local (peripheral) effects.

Aim

To elucidate the effects of serotonin (5-HT) and the SRIs clomipramine, fluoxetine and imipramine on the tension induced by norepinephrine (NE) of isolated human SV smooth muscle, as well as on the production of tissue cyclic AMP and cyclic GMP.

Main Outcome Measures

To measure the inhibition exerted by serotonin and SRIs clomipramine, fluoxetine, and imipramine on the contractile response of isolated SV tissue. In addition, the effects of the drugs on the turn-over of cyclic nucleotides cAMP and cGMP were also elucidated.

Methods

The effects of the cumulative addition of serotonin and the SRIs clomipramine, fluoxetine and imipramine (1 nM–10 µM) on the tension induced by the alpha1-adrenoceptor agonist NE (10 µM) of SV strip preparations were studied using the organ bath technique. Cyclic AMP and cyclic GMP were measured by means of specific radioimmunoassays.

Results

The tension induced by NE was dose-dependently reversed by the drugs tested. The rank order of efficacy was: imipramine ≥ fluoxetine ≥ clomipramine > serotonin. Mean reversion of tension was measured between 66 ± 6.6% and 52 ± 6.6%. These effects were paralleled by a 1.3-fold to 2.7-fold increase in tissue cAMP in response to exposure to the drugs. In contrast, no significant enhancement in cGMP was noted.

Conclusions

The findings, for the first time, present evidence that SRIs may antagonize the sympathetic contraction of SV smooth muscle via stimulation of tissue cyclic AMP.
Ten-Year Follow-Up of Sildenafil Use in Spinal Cord-Injured Patients with Erectile Dysfunction
Authors: Lombardi Giuseppe , Macchiarella Angelo , Cecconi Filippo , Del Popolo Giulio
Published: August 2009

Introduction

Data are sparse concerning the long-term effects of phosphodiesterase type 5 (PDE5) inhibitors for erectile dysfunction (ED).

Aim

To evaluate the efficacy and safety of long-term sildenafil use in subjects with ED caused by spinal cord injury (SCI).

Methods

Phase 1: From October 1998 to January 1999, 113 SCI patients with ED were given 50 mg of sildenafil after a 4-week treatment-free period. Those with a score lower than 26 on the International Index of Erectile Function (IIEF-15) and with less than 75% total successful sexual attempts the dosage of sildenafil was increased to 100 mg. Attempts were evaluated using the Sexual Encounter Profile Questions 2 and 3 (SEP2 and 3) regarding respectively the capacity to penetrate their partner and to maintain the erection after penetration. Phase 2: Only responding patients entered phase 2 where they were evaluated every 6 months. The final visit was concluded by January 2009.

Main Outcome Measures

Follow-up using the IIEF-15 questionnaire every 6 months.

Results

Seventy-five patients entered Phase 2. Thirty-eight patients were excluded, 35 of them because they did not respond to the drug. Lesions higher than T12, an incompleteness of lesions, and higher residual erection were significant predictable factors for the success of the therapy (P < 0.05). Phase 2: the most frequent reason (68.3%) for discontinuing treatment was the desire to try a new oral therapy especially for patients using 100 mg. Thirty-four individuals continued treatment, 28 of whom took 50 mg.

Conclusion

Sildenafil represents an effective and safe long-term option for SCI subjects with ED. Further investigation of long-term use of oral PDE5 inhibitors in SCI patients is needed for evaluating both factors that are determinant in the choice of a starter treatment and in detecting elements that influence the switching from initial treatment.
Variation in Susceptibility to Hurricane Damage as a Function of Storm Intensity in Puerto Rican Tree Species
Authors: Canham Charles D. , Thompson Jill , Zimmerman Jess K. , Uriarte María
Published: August 2009

One of the most significant challenges in developing a predictive understanding of the long‐term effects of hurricanes on tropical forests is the development of quantitative models of the relationships between variation in storm intensity and the resulting severity of tree damage and mortality. There have been many comparative studies of interspecific variation in resistance of trees to wind damage based on aggregate responses to individual storms. We use a new approach, based on ordinal logistic regression, to fit quantitative models of the susceptibility of a tree species to different levels of damage across an explicit range of hurricane intensity. Our approach simultaneously estimates both the local intensity of the storm within a plot and the susceptibility to storm damage of different tree species within plots. Using the spatial variation of storm intensity embedded in two hurricanes (Hugo in 1989 and Georges in 1998) that struck the 16 ha Luquillo Forest Dynamics Plot in eastern Puerto Rico, we show that variation in susceptibility to storm damage is an important aspect of life history differentiation. Pioneers such as Cecropia schreberiana are highly susceptible to stem damage, while the late successional species Dacryodes excelsa suffered very little stem damage but significant crown damage. There was a surprisingly weak relationship between tree diameter and the susceptibility to damage for most of the 12 species examined. This may be due to the effects of repeated storms and trade winds on the architecture of trees and forest stands in this Puerto Rican subtropical wet forest.
Crops and Livestock Productivity Growth in the Prairies: The Impacts of Technical Change and Scale
Authors: Stewart Bryce , Veeman Terrence , Unterschultz James
Published: August 2009

This study undertakes a comprehensive analysis of productivity growth in Canadian Prairie primary agriculture from 1940 to 2004. Total factor productivity (TFP) is measured using Törnqvist‐Theil indexing procedures for the Prairie provinces (Alberta, Manitoba, and Saskatchewan). During the 1940–2004 period, productivity growth in Prairie agriculture grew at a rate of 1.56% a year. This aggregate measure does not indicate the substantial variations in productivity growth that have occurred between crops and livestock, between the provinces, and over time: productivity growth in crops is considerably higher than productivity growth in livestock; Manitoba and Saskatchewan display consistently higher productivity growth than Alberta; and from 1980 to 2004 livestock productivity growth increased considerably in Manitoba and Saskatchewan. The productivity growth estimates are decomposed econometrically using a translog cost function to indicate the relative roles of technical change and scale effects. Productivity growth in crops has largely been the result of technical change while economies of scale have played a critical role in generating productivity growth in the livestock sector.

La présente étude est une analyse approfondie de la croissance de la productivité du secteur de l’agriculture primaire dans les provinces des Prairies (Alberta, Manitoba et Saskatchewan), de 1940 à 2004. Nous avons mesuré la productivité totale des facteurs (PTF) à l’aide de l’indice Törnqvist‐Theil. De 1940 à 2004, la productivité de l’agriculture dans les provinces des Prairies a cru au rythme annuel de 1,56 p. 100. Cette mesure globale n’indique pas les variations substantielles de la croissance de la productivité entre le secteur des cultures et le secteur de l’élevage, entre les provinces, et au fil du temps. La croissance de la productivité dans le secteur des cultures a été considérablement plus élevée que celle observée dans le secteur de l’élevage. La croissance de la productivité au Manitoba et en Saskatchewan a été supérieure à celle observée en Alberta. De 1980 à 2004, la croissance de la productivité du secteur de l’élevage a enregistré une hausse considérable au Manitoba et en Saskatchewan. Nous avons décomposé la croissance de la productivité de façon économétrique à l’aide d’une fonction de coût de type translog afin de faire ressortir le rôle des changements technologiques et des économies d’échelle. Les changements technologiques ont joué un rôle important dans la croissance de la productivité du secteur des cultures, tandis que les économies d’échelle ont joué un rôle primordial dans la croissance de la productivité du secteur de l’élevage.
INHIBITION OF TNFα PRODUCTION AND BLOCKING OF MITOGEN‐ACTIVATED PROTEIN KINASE/NFκB ACTIVATION IN LIPOPOLYSACCHARIDE‐INDUCED THP‐1 HUMAN MONOCYTES BY 3‐O‐ACETYL‐11‐KETO‐β‐BOSWELLIC ACID
Authors: SENGUPTA KRISHANU , GOLAKOTI TRIMURTULU , MARASETTI AJIT K. , TUMMALA TEJASWI , RAVADA SURYACHANDRA R. , KRISHNARAJU ALLURI V. , RAYCHAUDHURI SIBA P.
Published: August 2009

Boswellia serrataresin is regarded as a potent anti‐inflammatory agent in traditional and herbal medicine in the Indian subcontinent. The compound 3‐O‐acetyl‐11‐keto‐β‐boswellic acid (AKBA) is the most effective boswellic acid and mostly responsible forB. serrata's anti‐inflammatory properties. Here, we reexamined the anti‐inflammatory potential of a product selectively enriched with 30% AKBA (BE‐30, also known as 5‐Loxin®) and evaluated its underlying possible molecular mechanism of action. BE‐30 was 42.96% more effective than regularBoswelliaextract (BE‐3) in inhibiting 5‐lipoxygenase activity. In lipopolysaccharide (LPS)‐induced THP‐1 human monocytes, BE‐30 showed a strong anti‐TNFαactivity (half maximal inhibitory concentration 4.61 ± 0.87 µg/mL), which provides 71.14% (P < 0.001) better efficacy than BE‐3. Our investigations suggest that BE‐30 inhibits the LPS‐induced activation of serine/threonine kinases of mitogen‐activated protein kinase family, which are the key players responsible for a variety of cellular responses, including inflammation. Additionally, we also show that BE‐30 blocks the LPS‐induced NFκB activation by inhibiting IκBα phosphorylation and p65 translocation to the nuclear compartment of THP‐1 monocytes. Collectively, these findings provide molecular basis for the anti‐inflammatory properties of BE‐30.

PRACTICAL APPLICATIONS

This article describes the underlying molecular mechanisms for the anti‐inflammatory activities of an enriched formulation containing up to 30% 3‐O‐acetyl‐11‐keto‐β‐boswellic acid (AKBA), the active principle that is mainly responsible forBoswellia serrata'santi‐inflammatory properties. This AKBA‐enriched formulation (BE‐30), known as 5‐Loxin, is commercially available in the United States and is being used as a key ingredient of several formulations for improvement of joint health. This work explains anti‐TNFα properties of BE‐30 in a cellular inflammation modelin vitroand its inhibitory action on MAPK pathways in inflammation and, in addition, its anti‐NFκB activities. The findings will provide a further comprehensive mechanism of anti‐inflammatory action of 5‐Loxin at the cellular and molecular level.
EFFECT OF CHEMICAL INTERESTERIFICATION ON PHYSICOCHEMICAL PROPERTIES AND INDUSTRIAL APPLICATIONS OF CANOLA OIL AND FULLY HYDROGENATED COTTONSEED OIL BLENDS
Authors: RIBEIRO ANA PAULA BADAN , BASSO RODRIGO CORRÊA , GRIMALDI RENATO , GIOIELLI LUIZ ANTONIO , GONÇALVES LIRENY APARECIDA GUARALDO
Published: August 2009

Blends of canola oil (CO) and fully hydrogenated cottonseed oil (FHCSO), with 20, 25, 30, 35 and 40% FHCSO (w/w) were interesterified under the following conditions: 0.4% sodium methoxide, 500 rpm stirring, 100C, 20 min. The original and interesterified blends were examined for triacylglycerol composition, melting point, solid fat content (SFC) and consistency. Interesterification caused considerable rearrangement of triacylglycerol species, reduction of trisaturated triacylglycerol content and increase in disaturated‐monounsaturated and monosaturated‐diunsaturated triacylglycerols in all blends, resulting in lowering of respective melting points. The interesterified blends showed reduced SFC at all temperatures and more linear melting profiles if compared with the original blends. Consistency, expressed as yield value, significantly decreased after the reaction. Iso‐solid curves indicated eutectic interactions for the original blends, which were eliminated after randomization. The 80:20, 75:25, 70:30 and 65:35 (w/w) CO: FHCSO interesterified blends showed characteristics which are appropriate for their application as soft margarines, spreads, fat for bakery/all‐purpose shortenings, and icing shortenings, respectively.

PRACTICAL APPLICATIONS

Recently, a number of studies have suggested a direct relationship between trans isomers and increased risk of vascular disease. In response, many health organizations have recommended reducing consumption of foods containing trans fatty acids. In this connection, chemical interesterification has proven the main alternative for obtaining plastic fats that have low trans isomer content or are even trans isomer free. This work proposes to evaluate the chemical interesterification of binary blends of canola oil and fully hydrogenated cottonseed oil and the specific potential application of these interesterified blends in food products.
OMEGA‐3 POLYUNSATURATED FATTY ACIDS AFFECT LEPTIN RECEPTOR GENE EXPRESSION IN PITUITARY GH4C1 CELL LINE
Authors: DI BENEDETTO RITA , SALVATI SERAFINA , ATTORRI LUCILLA , DI BIASE ANTONELLA
Published: August 2009

Leptin (Ob) and Ob receptor (Ob‐Rb) genes are expressed in human and rodent anterior pituitary, and an autocrine/paracrine loop in the production and regulation of Ob and Ob‐Rb in this tissue has been suggested. As brain membranes contain large amounts of polyunsaturated fatty acids (PUFAs), and PUFA may affect the levels of leptin as well as the receptor's environment, a possible regulatory role of exogenous PUFA on Ob and Ob‐Rb expression in pituitary cell lines was investigated. GH4C1 cells were incubated with arachidonic (ARA), eicosapentaenoic (EPA) and docosahexaenoic (DHA) acids and fatty acid incorporation, viability, and gene expression were measured. The results showed that although fatty acids were incorporated in GH4C1 cells at a minimum level, positively affected cell viability without differences, among the different fatty acids. On the contrary, ARA, EPA and DHA showed different effects on Ob‐Rb gene regulation that was upregulated by EPA, downregulated by DHA and not affected by ARA. The effects of leptin co‐incubation are also reported and discussed.

PRACTICAL APPLICATIONS

Data from this paper consolidate the observations regarding the role of n‐3 polyunsaturated fatty acid in gene expression regulation, and thus they could support the recommendation to enrich diet with fish or fish oil with high PUFA content.
Improving Efficiency in Australian Local Government: Structural Reform as a Catalyst for Effective Reform
Authors: DOLLERY BRIAN , CRASE LIN , O'KEEFE SUE
Published: August 2009

Municipal amalgamation has been the main policy instrument of local government structural reform programmes in Australia for well over a century. However, council consolidation programs have not achieved the intended cost savings or improved service provision promised by advocates of this means of structural reorganisation. This paper considers whether the failure of municipal amalgamation processes to produce significant economic benefits necessarily implies that structural reform programs that invoke consolidation have no place in Australian local government policy. It is argued that ‘top‐down’ state government structural reform policy initiatives carrying the threat of amalgamation constitute an efficient mechanism for evoking optimal ‘bottom‐up’ structural change models.
Mapping the Contours of Race: Griffith Taylor's Zones and Strata Theory
Authors: WINLOW HEATHER
Published: August 2009

This article explores the ways in which cartography served as a tool to reinforce racial divisions in the context of late nineteenth and early twentieth century race science. Racial and anthropometric mapping was an endeavour in which both European and new world anthropologists and geographers were involved. The focus here is on the work of Thomas Griffith Taylor – regarded as one of the founders of modern geography in Australia – who deployed a number of cartographic techniques to reinforce his racial theorisations. This article explores Taylor's ‘zones and strata’ portrayal of racial evolution, and other geological‐ style maps of racial difference. These representations are investigated from two standpoints. Firstly, Taylor's theories are situated within the wider context of the Victorian tradition of classifying race, a tradition where physical race type was often correlated with moral and intellectual traits, and which was supported by the acceptance of environmental determinism within geographical circles. Secondly, his maps are considered from the perspective that, as J.B. Harley has argued, maps are social texts that contain power and, as such, can be deconstructed. Taylor's cartographic representations resulted from the manipulation of the internal elements of the map text, such as shading and projection, and were supported by the widely held belief that human racial groups could be delineated through physical anthropometry.
Prison foodservice in England*
Authors: Edwards John S. A. , Hartwell Heather J. , Schafheitle Joachim
Published: August 2009

Feeding a prison population presents a number of challenges. This paper draws on two major studies conducted in 16 prisons (12 male, two female and two young offenders' institutes) to examine the foodservice provision of prisoners in England. Both studies used observation, kitchen records and unstructured interviews to establish food preparation methods and procedures. The first study measured the intake of food provided in eight male prisons (n = 506) over a 24‐h period; in the second study, data were collected from eight prisons, four male, two female and two young offenders' institutes, for 3 days, from the cyclical menus. The institutions' recipes, methods and average portion sizes were used to calculate food and nutrient intake. Unstructured interviews were conducted with inmates and prison warders in both studies. Results show that prisoners are provided with a high standard of food and, with the exception of some nutrients, consume a healthy diet.
Overload‐induced skeletal muscle extracellular matrix remodelling and myofibre growth in mice lacking IL‐6
Authors: White J. P. , Reecy J. M. , Washington T. A. , Sato S. , Le M. E. , Davis J. M. , Wilson L. B. , Carson J. A.
Published: August 2009

Aim:  Overloading healthy skeletal muscle produces myofibre hypertrophy and extracellular matrix remodelling, and these processes are thought to be interdependent for producing muscle growth. Inflammatory cytokine interleukin‐6 (IL‐6) gene expression is induced in overloaded skeletal muscle, and the loss of this IL‐6 induction can attenuate the hypertrophic response to overload (OV). Although the OV induction of IL‐6 in skeletal muscle may be an important regulator of inflammatory processes and satellite cell proliferation, less is known about its role in the regulation of extracellular matrix remodelling. The purpose of the current study was to examine if OV‐induced extracellular matrix remodelling, muscle growth, and associated gene expression were altered in mice that lack IL‐6, when compared with wild‐type mice.

Methods:  Male C57/BL6 (WT) and C57/BL6 × IL‐6−/− (IL‐6−/−) mice (10 weeks of age) were assigned to either a sham control or synergist ablation OV treatments for 3, 21 or 56 days.

Result:  Plantaris muscle mass increased 59% in WT and 116% in IL‐6−/− mice after 21 day OV. Myofibre CSA was also increased by 21 day OV in both WT and IL‐6−/− mice. OV induced a twofold greater increase in the volume of non‐contractile tissue in IL‐6−/− muscle compared to WT. OV also induced a significantly greater accumulation of hydroxyproline and procollagen‐1 mRNA in IL‐6−/− muscle, when compared with WT muscle after 21 day OV. Transforming growth factor‐β and insulin‐like growth factor‐1 mRNA expression were also induced to a greater extent in IL‐6−/− muscle when compared with WT muscle after 21 day OV. There was no effect of IL‐6 loss on the induction of myogenin, and cyclin D1 mRNA expression after 3 day OV. However, MyoD mRNA expression in 3 day OV IL‐6−/− muscle was attenuated when compared with WT OV mice.

Conclusion:  IL‐6 appears to be necessary for the normal regulation of extracellular matrix remodelling during OV‐induced growth.
Risk factors for feline hyperthyroidism in the UK
Authors: Wakeling J. , Everard A. , Brodbelt D. , Elliott J. , Syme H.
Published: August 2009

Objectives:Previous studies of cats in the USA and New Zealand have identified a number of risk factors for the development of hyperthyroidism including feeding of canned cat food and being non‐purebred. The objective of this study was to examine these and other risk factors in cats from London, UK.

Methods:A questionnaire‐based case‐control study of hyperthyroidism in cats greater than eight years of age was undertaken. Cases and controls were recruited from two groups of first opinion clinics in London, UK (five locations in total). The two‐page questionnaire investigated details of lifestyle, diet and exposure to environmental chemicals. Data analysis included multivariable analysis of risk factors using binary logistic regression.

Results:One hundred and nine hyperthyroid cats and 196 control cats were surveyed. Increasing age, non‐pure breed, use of a litter box, more than 50 per cent wet food in the diet, a diet that included fish and exposure to food in a can were identified as risk factors for the development of hyperthyroidism using multivariable analysis.

ClinicalSignificance:Risk factors for hyperthyroidism in cats from the UK appear similar to those of other countries. Exposure to food packaged in a can was identified as the major risk factor for the development of hyperthyroidism.
Identification of direct transcriptional targets of V600EBRAF/MEK signalling in melanoma
Authors: Packer Leisl M , East Philip , Reis‐Filho Jorge S , Marais Richard
Published: August 2009

Oncogenic mutations in BRAF are common in melanoma and drive constitutive activation of the MEK/ERK pathway. To elucidate the transcriptional events downstream of V600EBRAF/MEK signalling we performed gene expression profiling of A375 melanoma cells treated with potent and selective inhibitors of V600EBRAF and MEK (PLX4720 and PD184352 respectively). Using a stringent Bayesian approach, we identified 69 transcripts that appear to be direct transcriptional targets of this pathway and whose expression changed after 6 h of pathway inhibition. We also identified several additional genes whose expression changed after 24 h of pathway inhibition and which are likely to be indirect transcriptional targets of the pathway. Several of these were confirmed by demonstrating their expression to be similarly regulated when BRAF was depleted using RNA interference, and by using qRT‐PCR in other BRAF mutated melanoma lines. Many of these genes are transcription factors and feedback inhibitors of the ERK pathway and are also regulated by MEK signalling in NRAS mutant cells. This study provides a basis for understanding the molecular processes that are regulated by V600EBRAF/MEK signalling in melanoma cells.
Antihypertensive Drugs and New‐Onset Diabetes: A Retrospective Longitudinal Cohort Study
Authors: Jong Gwo‐Ping , Chang Mu‐Hsin , Tien Liyun , Li Shu‐Yi , Liou Yi‐Sheng , Lung Chi‐Hsuan , Ma Tsochiang
Published: August 2009

Antihypertensive drugs have been linked to new‐onset diabetes (NOD); however, data on the effect of these drugs on the development of NOD in hypertensive patients has not been well determined. We aimed to investigate the association between antihypertensive drugs and NOD. This was a retrospective cohort study performed using data from claim forms provided to the central region branch of the Bureau of National Health Insurance in Taiwan from January 2002 to December 2007. Prescriptions for antihypertensive drugs before the index date were retrieved from a prescription database. We estimated the odds ratios (ORs) of NOD associated with antihypertensive drug use; nondiabetic subjects served as the reference group. A total of 4233 NOD cases were identified in 24,688 hypertensive patients during the study period. The risk of NOD after adjusting for sex and age was higher among users of diuretics (OR = 1.10, 95% confidence interval [CI]= 1.01–1.20), beta‐blockers (BBS; OR = 1.12, 95% CI = 1.04–1.21), and calcium channel blockers (CCBs; OR = 1.10, 95% CI = 1.02–1.18) than among nonusers. Patients who take angiotensin‐converting enzyme (ACE) inhibitors (OR = 0.92, 95% CI = 0.84–1.00), angiotensin receptor blockers (ARB; OR = 0.90, 95% CI = 0.81–0.98), or alpha‐blockers (OR = 0.88, 95% CI = 0.80–0.98) are at a lower risk of developing NOD than nonusers. Vasodilators were not associated with the risk of NOD. The results of this study suggest that hypertensive patients who take ACE inhibitors, ARBs, or alpha‐blockers are at a lower risk of NOD. Diuretics, BBs, and CCBs were associated with a significant increase in the risk of NOD.
Phylogeny and evolution of Perezia (Asteraceae: Mutisieae: Nassauviinae)
Authors: SIMPSON Beryl B. , ARROYO Mary T. K. , SIPE Sandra , DIAS de MORAES Marta , McDILL Joshua
Published: August 2009

Abstract  A molecular phylogenetic analysis of most of the species of Perezia reveals that, as traditionally defined, the genus is not monophyletic with two species more closely related to Nassauvia than to Perezia. In addition, our results show that Burkartia (Perezia) lanigera is related to Acourtia and is the only member of that clade in South America. The remaining species are monophyletic and show a pattern of an early split between a western temperate and an eastern subtropical clade of species. Within the western clade, the phylogeny indicates a pattern of diversification that proceeded from southern, comparatively low‐elevation habitats to southern high‐elevation habitats, and ultimately into more northern high‐elevation habitats. The most derived clades are found in the high central Andes, where significant radiation has occurred.
Structural changes in myosin motors and filaments during relaxation of skeletal muscle
Authors: Brunello E. , Fusi L. , Reconditi M. , Linari M. , Bianco P. , Panine P. , Narayanan T. , Piazzesi G. , Lombardi V. , Irving M.
Published: August 2009

Structural changes in myosin motors and filaments during relaxation from short tetanic contractions of intact single fibres of frog tibialis anterior muscles at sarcomere length 2.14 μm, 4°C were investigated by X‐ray diffraction. Force declined at a steady rate for several hundred milliseconds after the last stimulus, while sarcomere lengths remained almost constant. During this isometric phase of relaxation the intensities of the equatorial and meridional M3 X‐ray reflections associated with the radial and axial distributions of myosin motors also recovered at a steady rate towards their resting values, consistent with progressive net detachment of myosin motors from actin filaments. Stiffness measurements confirmed that the fraction of motors attached to actin declined at a constant rate, but also revealed a progressive increase in force per motor. The interference fine structure of the M3 reflection suggested that actin‐attached myosin motors are displaced towards the start of their working stroke during isometric relaxation. There was negligible recovery of the intensities of the meridional and layer‐line reflections associated with the quasi‐helical distribution of myosin motors in resting muscle during isometric relaxation, and the 1.5% increase in the axial periodicity of the myosin filament associated with muscle activation was not reversed. When force had decreased to roughly half its tetanus plateau value, the isometric phase of relaxation abruptly ended, and the ensuing chaotic relaxation had an exponential half‐time of ca 60 ms. Recovery of the equatorial X‐ray intensities was largely complete during chaotic relaxation, but the other X‐ray signals recovered more slowly than force.
Aryl Rhodanines Specifically Inhibit Staphylococcal and Enterococcal Biofilm Formation
Authors: Opperman Timothy J. , Kwasny Steven M. , Williams John D. , Khan Atiyya R. , Peet Norton P. , Moir Donald T. , Bowlin Terry L.
Published: August 2009

Staphylococcus epidermidis
and
Staphylococcus aureus
are the leading causative agents of indwelling medical device infections because of their ability to form biofilms on artificial surfaces. Here we describe the antibiofilm activity of a class of small molecules, the aryl rhodanines, which specifically inhibit biofilm formation of
S. aureus
,
S. epidermidis
,
Enterococcus faecalis
,
E. faecium
, and
E. gallinarum
but not the gram-negative species
Pseudomonas aeruginosa
or
Escherichia coli
. The aryl rhodanines do not exhibit antibacterial activity against any of the bacterial strains tested and are not cytotoxic against HeLa cells. Preliminary mechanism-of-action studies revealed that the aryl rhodanines specifically inhibit the early stages of biofilm development by preventing attachment of the bacteria to surfaces.
Antimicrobial Activities of Piperacillin-Tazobactam against
Haemophilus influenzae
Isolates, Including β-Lactamase-Negative Ampicillin-Resistant and β-Lactamase-Positive Amoxicillin- Clavulanate-Resistant Isolates, and Mutations in Their Quinolone Resistance-Determining Regions
Authors: Hirakata Yoichi , Ohmori Kaori , Mikuriya Miwako , Saika Takeshi , Matsuzaki Kaoru , Hasegawa Miyuki , Hatta Masumitsu , Yamamoto Natsuo , Kunishima Hiroyuki , Yano Hisakazu , Kitagawa Miho , Arai Kazuaki , Kawakami Kazuyoshi , Kobayashi Intetsu , Jones Ronald N. , Kohno Shigeru , Yamaguchi Keizo , Kaku Mitsuo
Published: August 2009

β-Lactamase-negative ampicillin-resistant (BLNAR) isolates of
Haemophilus influenza
e
have been emerging in some countries, including Japan. The Clinical and Laboratory Standards Institute has only a susceptible MIC breakpoint (≤1 μg/ml) for piperacillin-tazobactam and a disclaimer comment that BLNAR
H. influenzae
should be considered resistant, which was adapted without presentation of data. In addition, fluoroquinolone-resistant
H. influenzae
isolates have recently been occasionally reported worldwide. To address these problems, we examined susceptibilities to β-lactams, including piperacillin-tazobactam, and ciprofloxacin by microdilution and disk diffusion (only for piperacillin-tazobactam) methods, against a total of 400 recent
H. influenzae
clinical isolates, including 100 β-lactamase-negative ampicillin-susceptible, β-lactamase-positive ampicillin-resistant, BLNAR, and β-lactamase-positive amoxicillin-clavulanate-resistant (BLPACR) isolates each. BLNAR and BLPACR isolates were tested by PCR using primers that amplify specific regions of the
ftsI
gene. We also detected mutations in quinolone resistance-determining regions (QRDRs) by direct sequencing of the PCR products of DNA fragments. Among β-lactams, piperacillin-tazobactam exhibited potent activity against all isolates of
H. influenzae
, with all MICs at ≤0.5 μg/ml (susceptible). A disk diffusion breakpoint for piperacillin-tazobactam of ≥21 mm is proposed. We confirmed that all BLNAR and BLPACR isolates had amino acid substitutions in the
ftsI
gene and that the major pattern was group III-like (87.5%). One ciprofloxacin-resistant isolate (MIC, 16 μg/ml) and 31 ciprofloxacin-susceptible isolates (MICs, 0.06 to 0.5 μg/ml) had amino acid changes in their QRDRs. Piperacillin-tazobactam was the most potent β-lactam tested against all classes of
H. influenzae
isolates. It is possible that fluoroquinolone-resistant
H. influenzae
will emerge since several clinical isolates carried mutations in their QRDRs.
In Vitro Generation of Neuraminidase Inhibitor Resistance in A(H5N1) Influenza Viruses
Authors: Hurt Aeron C. , Holien Jessica K. , Barr Ian G.
Published: August 2009

To identify mutations that can arise in highly pathogenic A(H5N1) viruses under neuraminidase inhibitor selective pressure, two antigenically different strains were serially passaged with increasing levels of either oseltamivir or zanamivir. Under oseltamivir pressure, both A(H5N1) viruses developed a H274Y neuraminidase mutation, although in one strain the mutation occurred in combination with an I222M neuraminidase mutation. The H274Y neuraminidase mutation reduced oseltamivir susceptibility significantly (900- to 2,500-fold compared to the wild type). However the dual H274Y/I222M neuraminidase mutation had an even greater impact on resistance, with oseltamivir susceptibility reduced significantly further (8,000-fold compared to the wild type). A similar affect on oseltamivir susceptibility was observed when the dual H274Y/I222M mutations were introduced, by reverse genetics, into a recombinant seasonal human A(H1N1) virus and also when an alternative I222 substitution (I222V) was generated in combination with H274Y in A(H5N1) and A(H1N1) viruses. These viruses remained fully susceptible to zanamivir but demonstrated reduced susceptibility to peramivir. Following passage of the A(H5N1) viruses in the presence of zanamivir, the strains developed a D198G neuraminidase mutation, which reduced susceptibility to both zanamivir and oseltamivir, and also an E119G neuraminidase mutation, which demonstrated significantly reduced zanamivir susceptibility (1,400-fold compared to the wild type). Mutations in hemagglutinin residues implicated in receptor binding were also detected in many of the resistant strains. This study identified the mutations that can arise in A(H5N1) under either oseltamivir or zanamivir selective pressure and the potential for dual neuraminidase mutations to result in dramatically reduced drug susceptibility.
Natural Polymorphisms of Human Immunodeficiency Virus Type 1 Integrase and Inherent Susceptibilities to a Panel of Integrase Inhibitors
Authors: Low Andrea , Prada Nicole , Topper Michael , Vaida Florin , Castor Delivette , Mohri Hiroshi , Hazuda Daria , Muesing Mark , Markowitz Martin
Published: August 2009

We evaluated the human immunodeficiency virus type 1 (HIV-1) integrase coding region of thepolgene for the presence of natural polymorphisms in patients during early infection (AHI) and with triple-class drug-resistant HIV-1 (MDR). We analyzed selected recombinant viruses containing patient-derived HIV-1 integrase for susceptibility to a panel of strand transfer integrase inhibitors (InSTI). A pretreatment sequence analysis of the integrase coding region was performed for 112 patients identified during acute or early infection and 15 patients with triple-class resistance. A phenotypic analysis was done on 10 recombinant viruses derived from nine patients against a panel of six diverse InSTI. Few of the polymorphisms associated with in vitro InSTI resistance were identified in the samples from newly infected individuals or those patients with MDR HIV-1. We identified polymorphisms V72I, L74I, T97A, V151I, M154I/L, E157Q, V165I, V201I, I203M, T206S, and S230N. V72I was the most common, seen in 63 (56.3%) of the AHI samples. E157Q was the only naturally occurring mutation thought to contribute to resistance to elvitegravir, raltegravir, and L-870,810. None of the patient-derived viruses demonstrated any significant decrease in susceptibility to the drugs tested. In summary, the integrase coding region contains as much natural variation as that seen in protease, but mutations associated with high-level resistance to existing InSTI are rarely, if ever, present in integrase naïve patients, especially those being used clinically. Most of the highly prevalent polymorphisms have little effect on InSTI susceptibility in the absence of specific primary mutations. Baseline testing for integrase susceptibility in InSTI-naïve patients is not currently warranted.
IND-6, a Highly Divergent IND-Type Metallo-β-Lactamase fromChryseobacterium indologenesStrain 597 Isolated in Burkina Faso
Authors: Zeba Boukaré , De Luca Filomena , Dubus Alain , Delmarcelle Michael , Simporé Jacques , Nacoulma Odile G. , Rossolini Gian Maria , Frère Jean-Marie , Docquier Jean-Denis
Published: August 2009

The genusChryseobacteriumand other genera belonging to the familyFlavobacteriaceaeinclude organisms that can behave as human pathogens and are known to cause different kinds of infections. Several species ofFlavobacteriaceae, includingChryseobacterium indologenes, are naturally resistant to β-lactam antibiotics (including carbapenems), due to the production of a resident metallo-β-lactamase. AlthoughC. indologenespresently constitutes a limited clinical threat, the incidence of infections caused by this organism is increasing in some settings, where isolates that exhibit multidrug resistance phenotypes (including resistance to aminoglycosides and quinolones) have been detected. Here, we report the identification and characterization of a new IND-type variant from aC. indologenesisolate from Burkina Faso that is resistant to β-lactams and aminoglycosides. The levels of sequence identity of the new variant to other IND-type metallo-β-lactamases range between 72 and 90% (for IND-4 and IND-5, respectively). The purified enzyme exhibited N-terminal heterogeneity and a posttranslational modification consisting of the presence of a pyroglutamate residue at the N terminus. IND-6 shows a broad substrate profile, with overall higher turnover rates than IND-5 and higher activities than IND-2 and IND-5 against ceftazidime and cefepime.
Mycobacterium tuberculosisCell Wall Glycolipids Directly Inhibit CD4+T-Cell Activation by Interfering with Proximal T-Cell-Receptor Signaling
Authors: Mahon Robert N. , Rojas Roxana E. , Fulton Scott A. , Franko Jennifer L. , Harding Clifford V. , Boom W. Henry
Published: August 2009

Immune evasion is required forMycobacterium tuberculosisto survive in the face of robust adaptive CD4+T-cell responses. We have previously shown thatM. tuberculosiscan indirectly inhibit CD4+T cells by suppressing the major histocompatibility complex class II antigen-presenting cell function of macrophages. This study was undertaken to determine ifM. tuberculosiscould directly inhibit CD4+T-cell activation. Murine CD4+T cells were purified from spleens by negative immunoaffinity selection followed by flow sorting. Purified CD4+T cells were activated for 16 to 48 h with CD3 and CD28 monoclonal antibodies in the presence or absence ofM. tuberculosisand its subcellular fractions. CD4+T-cell activation was measured by interleukin 2 production, proliferation, and expression of activation markers, all of which were decreased in the presence ofM. tuberculosis. Fractionation identified thatM. tuberculosiscell wall glycolipids, specifically, phosphatidylinositol mannoside and mannose-capped lipoarabinomannan, were potent inhibitors. Glycolipid-mediated inhibition was not dependent on Toll-like receptor signaling and could be bypassed through stimulation with phorbol 12-myristate 13-acetate and ionomycin. ZAP-70 phosphorylation was decreased in the presence ofM. tuberculosisglycolipids, indicating thatM. tuberculosisglycolipids directly inhibited CD4+T-cell activation by interfering with proximal T-cell-receptor signaling.