bluuebunny/crossref_metadata_2025 huggingface dataset:
Silencing of WIPK and SIPK Mitogen-Activated Protein Kinases Reduces Tobacco mosaic virus Accumulation But Permits Systemic Viral Movement in Tobacco Possessing the N Resistance Gene
Authors: Kobayashi Michie , Seo Shigemi , Hirai Katsuyuki , Yamamoto-Katou Ayako , Katou Shinpei , Seto Hideharu , Meshi Tetsuo , Mitsuhara Ichiro , Ohashi Yuko
Published: July 2010

Infection of tobacco cultivars possessing the N resistance gene with Tobacco mosaic virus (TMV) results in confinement of the virus by necrotic lesions at the infection site. Although the mitogen-activated protein kinases WIPK and SIPK have been implicated in TMV resistance, evidence linking them directly to disease resistance is, as yet, insufficient. Viral multiplication was reduced slightly in WIPK- or SIPK-silenced plants but substantially in WIPK/SIPK-silenced plants, and was correlated with an increase in salicylic acid (SA) and a decrease in jasmonic acid (JA). Silencing of WIPK and SIPK in a tobacco cultivar lacking the N gene did not inhibit viral accumulation. The reduction in viral accumulation was attenuated by expressing a gene for an SA-degrading enzyme or by exogenously applying JA. Inoculation of lower leaves resulted in the systemic spread of TMV and formation of necrotic lesions in uninoculated upper leaves. These results suggested that WIPK and SIPK function to negatively regulate local resistance to TMV accumulation, partially through modulating accumulation of SA and JA in an N-dependent manner, but positively regulate systemic resistance.
A cbb3-Type Cytochrome C Oxidase Contributes to Ralstonia solanacearum R3bv2 Growth in Microaerobic Environments and to Bacterial Wilt Disease Development in Tomato
Authors: Colburn-Clifford Jennifer , Allen Caitilyn
Published: July 2010

Ralstonia solanacearum race 3 biovar 2 (R3bv2) is an economically important soilborne plant pathogen that causes bacterial wilt disease by infecting host plant roots and colonizing the xylem vessels. Little is known about R3bv2 behavior in the host rhizosphere and early in bacterial wilt pathogenesis. To explore this part of the disease cycle, we used a novel taxis-based promoter-trapping strategy to identify pathogen genes induced in the plant rhizosphere. This screen identified several rex (root exudate expressed) genes whose promoters were upregulated in the presence of tomato root exudates. One rex gene encodes an assembly protein for a high affinity cbb3-type cytochrome c oxidase (cbb3-cco) that enables respiration in low-oxygen conditions in other bacteria. R3bv2 cbb3-cco gene expression increased under low-oxygen conditions, and a cbb3-cco mutant strain grew more slowly in a microaerobic environment (0.5% O2). Although the cco mutant could still wilt tomato plants, symptom onset was significantly delayed relative to the wild-type parent strain. Further, the cco mutant did not colonize host stems or adhere to roots as effectively as wild type. These results suggest that R3bv2 encounters low-oxygen environments during its interactions with host plants and that the pathogen depends on this oxidase to help it succeed in planta.
Xanthomonas campestris pv. vesicatoria Effector AvrBsT Induces Cell Death in Pepper, but Suppresses Defense Responses in Tomato
Authors: Kim Nak Hyun , Choi Hyong Woo , Hwang Byung Kook
Published: July 2010

A type III effector protein, AvrBsT, is secreted into plant cells from Xanthomonas campestris pv. vesicatoria Bv5-4a, which causes bacterial spot disease on pepper (Capsicum annuum) and tomato (Solanum lycopersicum). To define the function and recognition of AvrBsT in the two host plants, avrBsT was introduced into the virulent pepper strain X. campestris pv. vesicatoria Ds1. Expression of AvrBsT in Ds1 rendered the strain avirulent to pepper plants. Infection of pepper leaves with Ds1 (avrBsT) expressing AvrBsT but not with near-isogenic control strains triggered a hypersensitive response (HR) accompanied by strong H2O2 generation, callose deposition, and defense-marker gene expressions. Mutation of avrBsT, however, compromised HR induction by X. campestris pv. vesicatoria Bv5-4a, suggesting its avirulence function in pepper plants. In contrast, AvrBsT acted as a virulence factor in tomato plants. Growth of strains Ds1 (avrBsT) and Bv5-4a ΔavrBsT was significantly enhanced and reduced, respectively, in tomato leaves. X. campestris pv. vesicatoria-expressed AvrBsT also significantly compromised callose deposition and defense-marker gene expression in tomato plants. Together, these results suggest that the X. campestris pv. vesicatoria type III effector AvrBsT is differentially recognized by pepper and tomato plants.
Sex differences in phenotypic plasticity of a mechanism that controls body size: implications for sexual size dimorphism
Authors: Stillwell R. Craig , Davidowitz Goggy
Published: July 2010

The degree and/or direction of sexual size dimorphism (SSD) varies considerably among species and among populations within species. Although this variation is in part genetically based, much of it is probably due to the sexes exhibiting differences in body size plasticity. Here, we use the hawkmoth,Manduca sexta, to test the hypothesis that moths reared on different diet qualities and at different temperatures will exhibit sex-specific body size plasticity. In addition, we explore the proximate mechanisms that potentially create sex-specific plasticity by examining three physiological variables known to regulate body size in this insect: the growth rate, the critical weight (which measures the cessation of juvenile hormone secretion from the corpora allata) and the interval to cessation of growth (ICG; which measures the time interval between the critical weight and the secretion of the ecdysteroids that regulate pupation and metamorphosis). We found that peak larval mass of males and females did not exhibit sex-specific plasticity in response to diet or temperature. However, the sexes did exhibit sex-specific plasticity in the mechanism that controls size; males and females exhibited sex-specific plasticity in the growth rate and the critical weight in response to both diet and temperature, whereas the ICG only exhibited sex-specific plasticity in response to diet. Our results suggest it is important for the sexes to maintain the same degree of SSD across environments and that this is accomplished by the sexes exhibiting differential sensitivity of the physiological factors that determine body size to environmental variation.
Diving through the thermal window: implications for a warming world
Authors: Campbell Hamish A. , Dwyer Ross G. , Gordos Matthew , Franklin Craig E.
Published: July 2010

Population decline and a shift in the geographical distribution of some ectothermic animals have been attributed to climatic warming. Here, we show that rises in water temperature of a few degrees, while within the thermal window for locomotor performance, may be detrimental to diving behaviour in air-breathing ectotherms (turtles, crocodilians, marine iguanas, amphibians, snakes and lizards). Submergence times and internal and external body temperature were remotely recorded from freshwater crocodiles (Crocodylus johnstoni) while they free-ranged throughout their natural habitat in summer and winter. During summer, the crocodiles' mean body temperature was 5.2 ± 0.1°C higher than in winter and the largest proportion of total dive time was composed of dive durations approximately 15 min less than in winter. Diving beyond 40 min during summer required the crocodiles to exponentially increase the time they spent on the surface after the dive, presumably to clear anaerobic debt. The relationship was not as significant in winter, even though a greater proportion of dives were of a longer duration, suggesting that diving lactate threshold (DLT) was reduced in summer compared with winter. Additional evidence for a reduced DLT in summer was derived from the stronger influence body mass exerted upon dive duration, compared to winter. The results demonstrate that the higher summer body temperature increased oxygen demand during the dive, implying that thermal acclimatization of the diving metabolic rate was inadequate. If the study findings are common among air-breathing diving ectotherms, then long-term warming of the aquatic environment may be detrimental to behavioural function and survivorship.
Identification of the cpsA gene as a specific marker for the discrimination of Streptococcus pneumoniae from viridans group streptococci
Authors: Park Hee Kuk , Lee Sang-Jae , Yoon Jang Won , Shin Jong Wook , Shin Hyoung-Shik , Kook Joong-Ki , Myung Soon Chul , Kim Wonyong
Published: July 2010

Streptococcus pneumoniae, the aetiological agent of pneumonia and non-gonococcal urethritis, shares a high degree of DNA sequence identity with the viridans group of streptococci, particularlyStreptococcus mitisandStreptococcus oralis. Although their clinical and pathological manifestations are different, discrimination betweenS. pneumoniaeand its close viridans cocci relatives is still quite difficult. Suppression subtractive hybridization was performed to identify the genomic differences betweenS. pneumoniaeandS. mitis. Thirty-four resultingS. pneumoniae-specific clones were examined by sequence determination and comparative DNA sequence analysis usingblast.S. pneumoniae-specific primers were subsequently designed from one of the clonal DNA sequences containing thecpsgene (coding for capsular polysaccharide biosynthesis). The primer specificities were evaluated using 49 viridans streptococci including 26S. pneumoniae, 54 other streptococci, 14Lactococcusspecies, 14Enterococcusspecies and threeVagococcusspecies, and compared with the specificities of previously described autolysin (lytA), pneumolysin (ply), Spn9802 and Spn9828 primers. The newly designedcpsA-specific primer set was highly specific toS. pneumoniaeand was even better than the existing primers. These findings may help improve the rapid identification and differentiation ofS. pneumoniaefrom closely related members of the viridans group streptococci.
Experimental Helicobacter marmotae infection in A/J mice causes enterohepatic disease
Authors: Patterson Mary M. , Rogers Arlin B. , Fox James G.
Published: July 2010

Helicobacter marmotaehas been identified in the inflamed livers of Eastern woodchucks (Marmota monax) infected with woodchuck hepatitis virus (WHV), as well as from the livers of WHV-negative woodchucks. Because the majority of WHV-positive woodchucks with hepatic tumours were culture or PCR positive for this helicobacter, and WHV-negative woodchucks withH. marmotaehad hepatitis, the bacterium may have a role in tumour promotion related to chronic inflammation. In this study, the type strain ofH. marmotaewas inoculated intraperitoneally into 48 male and female A/J mice, a strain noted to be susceptible toHelicobacter hepaticus-induced liver tumours. Sixteen mice served as mock-dosed controls. At 6, 12 and 18 months post-inoculation (p.i.), there were statistically significant (P<0.05) differences in mean inflammation scores for the caecum and proximal colon between experimentally infected and control mice. Differences in hepatic inflammation were significant (P<0.05) at 6 and 12 months p.i. between the two groups but not at the 18 month time point. Two infected male mice had livers with severe hepatitis, and the liver samples were culture positive forH. marmotae. Serum IgG levels in the mice dosed withH. marmotaewere elevated for the duration of the study. These results demonstrate that the woodchuck helicobacter can successfully colonize mice and cause enterohepatic disease. In the future, a mouse-adapted strain ofH. marmotaecould be selected to maximize colonization and lesion development. Such a woodchuck helicobacter-infected mouse model could be used to dissect potential mechanisms of microbial co-carcinogenesis involved in tumour development in woodchucks with WHV and in humans with hepatitis B virus.
Assembly of outer-membrane proteins in bacteria and mitochondria
Authors: Tommassen Jan
Published: July 2010

The cell envelope of Gram-negative bacteria consists of two membranes separated by the periplasm. In contrast with most integral membrane proteins, which span the membrane in the form of hydrophobicα-helices, integral outer-membrane proteins (OMPs) formβ-barrels. Similarβ-barrel proteins are found in the outer membranes of mitochondria and chloroplasts, probably reflecting the endosymbiont origin of these eukaryotic cell organelles. How theseβ-barrel proteins are assembled into the outer membrane has remained enigmatic for a long time. In recent years, much progress has been reached in this field by the identification of the components of the OMP assembly machinery. The central component of this machinery, called Omp85 or BamA, is an essential and highly conserved bacterial protein that recognizes a signature sequence at the C terminus of its substrate OMPs. A homologue of this protein is also found in mitochondria, where it is required for the assembly ofβ-barrel proteins into the outer membrane as well. Although accessory components of the machineries are different between bacteria and mitochondria, a mitochondrialβ-barrel OMP can be assembled into the bacterial outer membrane and, vice versa, bacterial OMPs expressed in yeast are assembled into the mitochondrial outer membrane. These observations indicate that the basic mechanism of OMP assembly is evolutionarily highly conserved.
Influence of Particle Diameter on Surface Silanol Structure, Hydration Forces, and Aggregation Behavior of Alkoxide‐Derived Silica Particles
Authors: Kamiya Hidehiro , Mitsui Mika , Takano Hideo , Miyazawa Sakura
Published: July 2010

The effects of silica particle diameter on dispersion and aggregation behavior in water were analyzed, using alkoxide‐derived silica powders with particle diameters of 8–260 nm. The present study focused on the relationships between the surface silanol structure and the interaction forces between solid surfaces in water. The surface silanol structure and interaction between particles were determined using Fourier transform infrared spectroscopy, Fourier transform near‐infrared spectroscopy, and atomic force microscopy. For relatively large particles (>30 nm in diameter), the surface silanols primarily were hydrogen‐bonded silanols, and isolated silanols disappeared. The hydrogen‐bonded silanols formed a hydrogen‐bonded water layer on the particle surface; therefore, the additional hydration force was strong between these relatively large particles. In contrast, the surface density of isolated silanols increased as the particle diameter decreased to <30 nm, and the additional hydration force between ultrafine powders disappeared. The aggregation behaviors of alkoxide‐derived silica powders were dependent on the hydration force, which was changed by the surface silanol structure.
US state alcohol sales compared to survey data, 1993–2006
Authors: Nelson David E. , Naimi Timothy S. , Brewer Robert D. , Roeber James
Published: July 2010

Aims Assess long‐term trends of the correlation between alcohol sales data and survey data.

Design Analyses of state alcohol consumption data from the US Alcohol Epidemiologic Data System based on sales, tax receipts or alcohol shipments. Cross‐sectional, state annual estimates of alcohol‐related measures for adults from the US Behavioral Risk Factor Surveillance System using telephone surveys.

Setting United States.

Participants State alcohol tax authorities, alcohol vendors, alcohol industry (sales data) and randomly selected adults aged ≥ 18 years 1993–2006 (survey data).

Measurements State‐level per capita annual alcohol consumption estimates from sales data. Self‐reported alcohol consumption, current drinking, heavy drinking, binge drinking and alcohol‐impaired driving from surveys. Correlation coefficients were calculated using linear regression models.

Findings State survey estimates of consumption accounted for a median of 22% to 32% of state sales data across years. Nevertheless, state consumption estimates from both sources were strongly correlated with annual r‐values ranging from 0.55–0.71. State sales data had moderate‐to‐strong correlations with survey estimates of current drinking, heavy drinking and binge drinking (range of r‐values across years: 0.57–0.65; 0.33–0.70 and 0.45–0.61, respectively), but a weaker correlation with alcohol‐impaired driving (range of r‐values: 0.24–0.56). There were no trends in the magnitude of correlation coefficients.

Conclusions Although state surveys substantially underestimated alcohol consumption, the consistency of the strength of the association between sales consumption and survey data for most alcohol measures suggest both data sources continue to provide valuable information. These findings support and extend the distribution of consumption model and single distribution theory, suggesting that both sales and survey data are useful for monitoring population changes in alcohol use.
Development of target‐specific treatments in multiple myeloma
Authors: Chanan‐Khan Asher A. , Borrello Ivan , Lee Kelvin P. , Reece Donna E.
Published: July 2010

In the last decade, the novel agents lenalidomide, bortezomib, and thalidomide have dramatically improved outcomes for patients with multiple myeloma (MM). A number of new therapies with precise targets involved in MM cell growth and replication are now in development and have the potential for further improvements. Second‐generation proteasome inhibitors and thalidomide derivatives may offer increased efficacy and safety. Investigational therapies with rationally selected targets in MM include inhibitors of histone deacetylase, heat shock protein 90, mammalian target of rapamycin, BCL2, Akt, mitogen‐activated protein kinase, and telomerase. In addition, monoclonal antibodies directed against several targets have been developed and many are showing promise in initial clinical trials in MM. Interest in the ancient remedy of arsenic trioxide has been revived because of its proapoptotic effects on mitochondria, despite its established toxicities. In general, combination regimens are proving the most efficacious, which is to be expected given the multiple overlapping pathways responsible for MM growth and progression.
Temporal and spatial monitoring of mobile nanoparticles in a vineyard soil: evidence of nanoaggregate formation
Authors: Perdrial N. , Perdrial J. N. , Delphin J.‐E. , Elsass F. , Liewig N.
Published: July 2010

Mechanisms of formation, stabilization, liberation, transport and deposition of nanoparticles and their relationship to contaminant transport remain scarcely investigated in natural porous media. This study investigated nanoparticles mobilized in the pore space of a French vineyard soil by observing mobile soil‐derived organic matter (SOM) and minerals in pore fluids over an 8‐month monitoring period. Samples were collected in situ and investigated by transmission electron microscopy coupled to electron‐dispersive spectroscopy. The main types of nanoparticles transported within the soil were clay, bacteria, SOM and nanoaggregates. Nanometric clay particles were enriched in various metals (Fe, Zn, As and Pb) and organically‐derived constituents. Analyses of bacteria showed enrichments in Pb. SOM consisted of small carbon‐based particles (<200 nm) with slight enrichments in various metals. The fourth dominant particle type consisted of the association of particles forming organo‐mineral nanoaggregates. Based on the study of more than 22 500 individual particles, we propose a schematic interpretation of the evolution of the distribution of particles with depth in a soil profile. The increase of nanoaggregates with depth in the soil seemed to be largely controlled by the ionic strength of soil water and soil hydrodynamics. Seasonal variations in temperature also appear to affect nanoaggregation. Based on the architecture of the nanoaggregates, we propose an improvement of pre‐existing models of microaggregation by focusing on early aggregation stages suggesting the importance of bacteria and electrostatic interactions. The process of nanoaggregation can enhance the net reactivity of soil with respect to transported suspended matter, including heavy metals, and can initiate the process of C sequestration.
Pulmonary adenocarcinoma with microcystic histology and intratumoral heterogeneity of EGFR gene polymorphism
Authors: Yeh Yi‐Chen , Chou Teh‐Ying
Published: July 2010

Yeh Y‐C & Chou T‐Y (2010) Histopathology 57, 112–120 Pulmonary adenocarcinoma with microcystic histology and intratumoral heterogeneity of EGFR gene polymorphism

Aims:  To study the clinicopathological features of pulmonary adenocarcinomas with microcystic histology.

Materials and results:  Two cases of pulmonary adenocarcinoma exhibiting a unique microcystic growth pattern were analysed. The 20‐mm tumour of a 59‐year‐old woman showed a predominantly complex microcystic architecture with a focal papillary configuration, whereas the 15‐mm tumour of another 46‐year‐old woman displayed a solely microcystic growth pattern. The first tumour was found to have metastasized to lymph nodes at surgery and, 4 years later, to the chest wall. The second tumour did not metastasize or recur over a follow‐up period of 12 years. The tumour cells were immunoreactive with thyroid transcription factor‐1 (TTF‐1), which helped to differentiate them from salivary gland‐type tumours. No mutations of epidermal growth factor receptor (EGFR) or K‐ras genes were detected. However, facilitated by laser microdissection, a Q787Q polymorphism was found in the microcystic component but not in the papillary component. To our knowledge, this is the first report to illustrate intratumoral heterogeneity of EGFR gene polymorphism among different growth patterns within the same tumour.

Conclusions:  Pulmonary microcystic adenocarcinoma has a variable clinical outcome. The microcystic tumour cells are immunoreactive with TTF‐1 and may harbour polymorphisms of the EGFR gene.
Use of isogenic strains indicates CYP9M10 is linked to permethrin resistance in Culex pipiens quinquefasciatus
Authors: Hardstone M. C. , Komagata O. , Kasai S. , Tomita T. , Scott J. G.
Published: July 2010

Previous studies on a strain of Culex pipiens quinquefasciatus from Saudi Arabia indicated permethrin resistance was a result of cytochrome P450 mediated detoxification and kdr. The P450 detoxification was found to be larval specific and associated with a fitness cost in certain environments. The P450 responsible for resistance (and the fitness cost) has not been identified, but recently two candidate P450s (CYP4H34 and CYP9M10) have been found. We measured cytochrome P450 and cytochrome b5 content as well as the expression levels of CYP4H34 and CYP9M10 in a susceptible (SLAB) and two isogenic strains (isolated by repeated backcrossing and selection) of mosquito (ISOP450 and ISOJPAL) resistant to permethrin. Cytochrome P450 protein levels of the resistant strains were significantly higher (1.5‐fold) than SLAB, but were not significantly different from one another. Expression of CYP4H34 in the larvae and adults of the resistant (ISOP450 and ISOJPAL) and susceptible (SLAB) strains were not statistically different. CYP9M10 was found to be significantly over‐expressed in larvae of both permethrin‐resistant isogenic strains (1800‐fold in ISOP450 and 870‐fold in ISOJPAL) when compared to SLAB. Partial sequence analysis of CYP9M10 revealed eight polymorphic sites that distinguished the susceptible allele from the resistant allele. We conclude that CYP9M10 is linked to permethrin resistance in these strains of C. p. quinquefasciatus, and is likely to be the P450 gene responsible for resistance in these strains.
Community adjustment of mobile people with learning difficulties after total hospital closure: a preliminary report
Authors: Brook S. Lister , Bowler D. M.
Published: July 2010

A study is described in which 50 residents of a hospital for people with leaming difficulties were assessed immediately before and i year after moving into two types of community provision; purpose‐built campuses or small homes in residential streets. Measures were taken of residents’ adaptive and unwanted behaviours and of aspects of their lifestyles. The results report the changes in these measures that were found for both campus and small home residents.
Assessing clinicians’ consultation with people with profound learning disability: producing a rating scale
Authors: Kerr M. P. , Evans S. , Nolan M. , Fraser W. I.
Published: July 2010

This paper describes the development of a scale for assessing clitiicians’ communication with people with profound learning disability. Four psychiatrists and four nurses were assessed completing three simple non‐invasive clinical procedures—blood pressure, pulse and axillary temperature—with people with profound learning disability. Videotaped assessmetit of consultations was performed by three experienced speech and language therapists using a previously designed scale for analysing encounters with people with mild learning disability. This led to the production of a new scale specifically for people with profound learning disability. A significant inter‐rater reliability was found between the three speech therapists for total scores (rater a‐b, corr = 0·654, P= 0·006; rater a–c, corr = 0·795, P= 0·0001: rater b–c, corr = 0·673, P = 0·004). Significant reliability between raters was also found for the subsections of verbal behaviour and non‐verbal behaviour.
The effect of age on language in people with Down's syndrome
Authors: Cooper S.‐A. , Collacott R. A.
Published: July 2010

It has been previously suggested, in a small‐sized study of 60 people with Down's syndrome, that receptive language skills were lower, the older the individual, whereas expressive language skills did not correlate with age (Carter‐Young & Kramer 1991), By assessing language skills on 345 Down's syndrome adults living in Leicestershire (91% of the total population), the present authors have been able to confirm this finding, using the AAMD Adaptive Behaviour Scale. This may have clinical relevance with regard to communication with older people with Down's syndrome, and also for estimation of an individual's skill level when he or she was younger.
Intra‐individual factors influencing efficacy of interventions for stereotyped behaviours: a meta‐analysis
Authors: Wehmeyer M. L.
Published: July 2010

The efficacy of treatments targeting stereotyped behaviours has been the focal point of several reviews. This study examined efficacy as a function of intra‐individual characteristics, including age, gender, level of disability and topography of the behaviour. A meta‐analysis of studies reporting treatments of stereotyped behaviours appearing in 23 journals over 20 years was conducted. Efficacy was determined using two metrics. Percentage Non‐overlapping Data and Percentage Zero Data, which indicate treatment success as a function of the degree to which the behaviour fell below baseline levels, and once reaching zero, remained there. Non‐parametric analyses indicated differences in treatment efficacy according to age and the form of the stereotypy. Implications for the design of treatments to address stereotypies are discussed.
Exercise effects on night‐to‐night fluctuations in self‐rated sleep among older adults with sleep complaints
Authors: BUMAN MATTHEW P. , HEKLER ERIC B. , BLIWISE DONALD L. , KING ABBY C.
Published: July 2010

Sleep interventions have rarely explored reductions in night‐to‐night fluctuations [i.e. intra‐individual variability (IIV)] in sleep, despite the negative impacts of such fluctuations on affective states and cognitive and physical symptoms. In a community‐based randomized controlled trial we evaluated whether physical exercise reduced IIV in self‐rated sleep outcomes among middle‐aged and older adults with sleep complaints. Under‐active adults 55 years and older (n = 66, 67% women) with mild to moderate sleep complaints were randomized to 12 months of a moderate‐intensity endurance exercise (n = 36) or a health education control group (n = 30). Daily sleep logs, Pittsburgh Sleep Quality Index (PSQI) and in‐home polysomnographic sleep recordings (PSG) were collected at baseline, 6 months and 12 months. Sleep log‐derived means and IIV were computed for sleep‐onset latency (SOL), time in bed, feeling rested in the morning, number of nighttime awakenings, and wake after final awakening (WAFA). Using intent‐to‐treat methods, at 6 months no differences in IIV were observed by group. At 12 months, SOL‐based IIV was reduced in the exercise group compared with the control (difference = 23.11, 95% CI: 3.04–47.18, P = 0.025, Cohen’s d = 0.57). This change occurred without mean‐level or IIV changes in sleep–wake schedules. For all sleep variables, except SOL and WAFA, IIV changes and mean‐level changes in each variable were negatively correlated (r = −0.312 to −0.691, P < 0.05). Sleep log‐derived IIV changes were modestly correlated with mean‐level PSQI and PSG‐based changes at 12 months. Twelve months of moderate‐intensity exercise reduced night‐to‐night fluctuations in self‐rated time to fall asleep, and this relationship was independent of mean‐level time to fall asleep.
Pharmacokinetics of cefpodoxime in plasma and subcutaneous fluid following oral administration of cefpodoxime proxetil in male beagle dogs
Authors: KUMAR V. , MADABUSHI R. , LUCCHESI M. B. B. , DERENDORF H.
Published: July 2010

Kumar, V., Madabushi, R., Lucchesi, M. B. B., Derendorf, H. Pharmacokinetics of cefpodoxime in plasma and subcutaneous fluid following oral administration of cefpodoxime proxetil in male beagle dogs. J. vet. Pharmacol. Therap.34, 130–135.

Pharmacokinetics of cefpodoxime in plasma (total concentration) and subcutaneous fluid (free concentration using microdialysis) was investigated in dogs following single oral administration of prodrug cefpodoxime proxetil (equivalent to 5 and 10 mg/kg of cefpodoxime). In a cross over study design, six dogs per dose were utilized after a 1 week washout period. Plasma, microdialysate, and urine samples were collected upto 24 h and analyzed using high performance liquid chromatography. The average maximum concentration (Cmax) of cefpodoxime in plasma was 13.66 (±6.30) and 27.14 (±4.56) μg/mL with elimination half‐life (t1/2) of 3.01 (±0.49) and 4.72 (±1.46) h following 5 and 10 mg/kg dose, respectively. The respective average area under the curve (AUC0–∞) was 82.94 (±30.17) and 107.71 (±30.79) μg·h/mL. Cefpodoxime was readily distributed to skin and average free Cmax in subcutaneous fluid was 1.70 (±0.55) and 3.06 (±0.93) μg/mL at the two doses. Urinary excretion (unchanged cefpodoxime) was the major elimination route. Comparison of subcutaneous fluid concentrations using pharmacokinetic/pharmacodynamic indices of fT>MIC indicated that at 10 mg/kg dose; cefpodoxime would yield good therapeutic outcome in skin infections for bacteria with MIC50 upto 0.5 μg/mL while higher doses (or more frequent dosing) may be needed for bacteria with higher MICs. High urine concentrations suggested cefpodoxime use for urinary infections in dogs.
The Rad4 homologue YDR314C is essential for strand‐specific repair of RNA polymerase I‐transcribed rDNA in Saccharomyces cerevisiae
Authors: den Dulk Ben , Brandsma Jourica A. , Brouwer Jaap
Published: July 2010

The Saccharomyces cerevisiae protein Rad4 is involved in damage recognition in nucleotide excision repair (NER). In RNA polymerase II‐transcribed regions Rad4 is essential for both NER subpathways global genome repair (GGR) and transcription coupled repair (TCR). In ribosomal DNA (rDNA), however, the RNA polymerase I‐transcribed strand can be repaired in the absence of Rad4. In Saccharomyces cerevisiae the YDR314C protein shows homology to Rad4. The possible involvement of YDR314C in NER was studied by analysing strand‐specific cyclobutane pyrimidine dimer (CPD) removal in both RNA pol I‐ and RNA pol II‐transcribed genes. Here we show that the Rad4‐independent repair of rDNA is dependent on YDR314C. Moreover, in Rad4 proficient cells preferential repair of the transcribed strand of RNA pol I‐transcribed genes was lost after deletion of YDR314C, demonstrating that Rad4 cannot replace YDR314C. CPD removal from the RNA pol II‐transcribed RPB2 gene was unaffected in ydr314c mutants. We conclude that the two homologous proteins Rad4 and YDR314C are both involved in NER and probably have a similar function, but operate at different loci in the genome and are unable to replace each other.
2‐Oxoglutarate and the PII homologues NifI1 and NifI2 regulate nitrogenase activity in cell extracts of Methanococcus maripaludis
Authors: Dodsworth Jeremy A. , Cady Nathaniel C. , Leigh John A.
Published: July 2010

Post‐translational regulation of nitrogen fixation, or switch‐off, in the methanogenic archaeon Methanococcus maripaludis does not involve detectable covalent modification of the dinitrogenase reductase as in some bacteria, and the genes encoding the PII homologues NifI1 and NifI2 are both required, indicating a novel mechanism. To further understand the mechanism of switch‐off, we assayed nitrogenase activity in cell extracts from wild‐type and nifI mutant strains in the absence or presence of potential signals of nitrogen status. Activity in extracts from a ΔnifI1nifI2 strain was sixfold higher than in extracts from wild‐type cells. Addition of 2‐oxoglutarate to wild‐type extracts enhanced activity up to fivefold, a level similar to that observed  in ΔnifI1nifI2  extracts.  2‐Oxoglutarate  did not affect activity in ΔnifI1nifI2 or single nifI mutant extracts. Furthermore, extracts from genetically complimented nifI mutants regained wild‐type characteristics, indicating an in vitro correlation with in vivo effects. Extraction and quantification of 2‐oxoglutarate indicated concentrations 10‐fold higher in nitrogen‐fixing cells than in switched‐off and ammonium‐grown cells. We propose a model for switch‐off where the NifI proteins have an inhibitory effect on nitrogenase activity  that  is  counteracted  by  high  levels  of 2‐oxoglutarate, which acts as a signal of nitrogen limitation.
Extent of the activity domain and possible roles of FtsK in the Escherichia coli chromosome terminus
Authors: Corre Jacqueline , Louarn Jean‐Michel
Published: July 2010

Escherichia coli FtsK protein couples cell division and chromosome segregation. It is a component of the septum essential for cell division. It also acts during chromosome dimer resolution by XerCD‐specific recombination at the dif site, with two distinct activities: DNA translocation oriented by skewed sequence elements and direct activation of Xer recombination. Dimer resolution requires that the skewed elements polarize in opposite directions 30–50 kb on either side of dif. This constitutes the DIF domain, approximately coincident with the region where replication terminates. The observation that the ftsK1 mutation increases recombination near dif was exploited to determine whether the chromosome region on which FtsK acts is limited to the DIF domain. A monitoring of recombination activity at multiple loci in a 350 kb region to the left of dif revealed (i) zones of differing activities unconnected to dimer resolution and (ii) a constant 10‐fold increase of recombination in the 250 kb region adjacent to dif in the ftsK1 mutant. The latter effect allows definition of an FTSK domain whose total size is at least fourfold that of the DIF domain. Additional analyses revealed that FtsK activity responds to polarization in the whole FTSK domain and that displacement of the region where replication terminates preserves differences between recombination zones. Our interpretation is that translocation by FtsK occurs mostly on DNA belonging to a specifically organized domain of the chromosome, when physical links between either dimeric or still intercatenated chromosomes force this DNA to run across the septum at division.
Elaborate transcription regulation of the Bacillus subtilis ilv‐leu operon involved in the biosynthesis of branched‐chain amino acids through global regulators of CcpA, CodY and TnrA
Authors: Tojo Shigeo , Satomura Takenori , Morisaki Kaori , Deutscher Josef , Hirooka Kazutake , Fujita Yasutaro
Published: July 2010

The Bacillus subtilis ilv‐leu operon involved in the biosynthesis of branched‐chain amino acids is under negative regulation mediated by TnrA and CodY, which recognize and bind to their respective cis‐elements located upstream of the ilv‐leu promoter. This operon is known to be under CcpA‐dependent positive regulation. We have currently identified a catabolite‐responsive element (cre) for this positive regulation (bases −96 to −82; +1 is the ilv‐leu transcription initiation base) by means of DNase I‐footprinting in vitro, and deletion and base‐substitution analyses of cre. Under nitrogen‐rich growth conditions in glucose‐minimal medium supplemented with glutamine and amino acids, CcpA and CodY exerted positive and negative regulation of ilv‐leu, respectively, but TnrA did not function. Moreover, CcpA and CodY were able to function without their counteracting regulation of each other, although the CcpA‐dependent positive regulation did not overcome the CodY‐dependent negative regulation. Furthermore, under nitrogen‐limited conditions in glucose‐minimal medium with glutamate as the sole nitrogen source, CcpA and TnrA exerted positive and negative regulation, respectively, but CodY did not function. This CcpA‐dependent positive regulation occurred without the TnrA‐dependent negative regulation. However, the TnrA‐dependent negative regulation did not occur without the CcpA‐dependent positive regulation, raising the possibility that this negative regulation might decrease the CcpA‐dependent positive regulation. The physiological role of this elaborate transcription regulation of the B. subtilis ilv‐leu operon in overall metabolic regulation in this organism is discussed.
Aerobic benzoyl‐CoA catabolic pathway in Azoarcus evansii: studies on the non‐oxygenolytic ring cleavage enzyme
Authors: Gescher Johannes , Eisenreich Wolfgang , Wörth Jürgen , Bacher Adelbert , Fuchs Georg
Published: July 2010

A novel aerobic benzoate pathway has recently been discovered in various bacteria in which benzoate is first converted to benzoyl‐CoA. The further downstream steps are associated with the gene products of the benzoate oxidation gene cluster (box) on the Azoarcus evansii chromosome. Benzoyl‐CoA is oxidized to 2,3‐dihydro‐2,3‐dihydroxybenzoyl‐CoA (benzoyl‐CoA dihydrodiol) by benzoyl‐CoA oxygenase/reductase BoxBA in the presence of molecular oxygen. This study identified the next, ring cleaving step catalysed by BoxC. The boxC gene was expressed in a recombinant Escherichia coli strain as a fusion protein with maltose binding protein (BoxCmal) and the wild type as well as the recombinant proteins were purified and studied. BoxC catalyses the reaction 2,3‐dihydro‐2,3‐dihydroxybenzoyl‐CoA + H2O → 3,4‐dehydroadipyl‐CoA semialdehyde + HCOOH. This is supported by the following results. Assays containing [ring‐13C6]benzoyl‐CoA, benzoyl‐CoA oxygenase/reductase, BoxCmal protein, NADPH and semicarbazide were analysed directly by NMR spectroscopy and mass spectrometry. The products were identified as the semicarbazone of [2,3,4,5,6‐13C5]3,4‐dehydroadipyl‐CoA semialdehyde; the missing one‐carbon unit being formate. The same reaction mixture without semicarbazide yielded a mixture of the hydrate of [2,3,4,5,6‐13C5]3,4‐dehydroadipyl‐CoA semialdehyde and [2,3,4,5,6‐13C5]4,5‐dehydroadipyl‐CoA semialdehyde. BoxC, a 122 kDa homodimeric enzyme (61 kDa subunits), is termed benzoyl‐CoA‐dihydrodiol lyase. It contains domains characteristic for enoyl‐CoA hydratases/isomerases, besides a large central domain with no significant similarity to sequences in the database. The purified protein did not require divalent metals, molecular oxygen or any cosubstrates or coenzymes for activity. The complex reaction is part of a widely distributed new principle of aerobic aromatic metabolism in which all intermediates are coenzyme A thioesters and the actual ring‐cleavage reaction does not require molecular oxygen.
Host factors that promote transpososome disassembly and the PriA‐PriC pathway for restart primosome assembly
Authors: North Stella H. , Nakai Hiroshi
Published: July 2010

Initiation of bacteriophage Mu DNA replication by transposition requires the disassembly of the transpososome that catalyses strand exchange and the assembly of a replisome promoted by PriA, PriB, PriC and DnaT proteins, which function in the host to restart stalled replication forks. Once the molecular chaperone ClpX weakens the very tight binding of the transpososome to the Mu ends, host disassembly factors (MRFα‐DF) promote the dissociation of the transpososome from the DNA template and the assembly of a new nucleoprotein complex. Prereplisome factors (MRFα‐PR) further alter the complex, allowing PriA binding and loading of major replicative helicase DnaB onto the template promoted by the restart proteins. MRFα‐PR is essential for DnaB loading by restart proteins even on the deproteinized Mu fork whereas MRFα‐DF is not required on the deproteinized template. When the transition from transpososome to replisome was reconstituted using MRFα‐DF and MRFα‐PR, initiation of Mu DNA replication was strictly dependent upon added PriC and PriA helicase. In contrast, initiation on the deproteinized template was predominantly dependent upon PriB and did not require PriA's helicase activity. The results indicate that transition mechanisms beginning with the transpososome disassembly can determine the pathway of replisome assembly by restart proteins.
Partition‐associated incompatibility caused by random assortment of pure plasmid clusters
Authors: Ebersbach Gitte , Sherratt David J. , Gerdes Kenn
Published: July 2010

Bacterial plasmids and chromosomes encode centromere‐like partition loci that actively segregate DNA before cell division. The molecular mechanism behind DNA segregation in bacteria is largely unknown. Here we analyse the mechanism of partition‐associated incompatibility for plasmid pB171, a phenotype associated with all known plasmid‐encoded centromere loci. An R1 plasmid carrying par2 from plasmid pB171 was destabilized by the presence of an F plasmid carrying parC1, parC2 or the entire par2 locus of pB171. Strikingly, cytological double‐labelling experiments revealed no evidence of long‐lived pairing of plasmids. Instead, pure R1 and F foci were positioned along the length of the cell, and in a random order. Thus, our results raise the possibility that partition‐mediated plasmid incompatibility is not caused by pairing of heterologous plasmids but instead by random positioning of pure plasmid clusters along the long axis of the cell. The strength of the incompatibility was correlated with the capability of the plasmids to compete for the mid‐cell position.
Inhibitory effect of ACC deaminase‐producing bacteria on crown gall formation in tomato plants infected by Agrobacterium tumefaciens or A. vitis
Authors: Toklikishvili N. , Dandurishvili N. , Vainstein A. , Tediashvili M. , Giorgobiani N. , Lurie S. , Szegedi E. , Glick B. R. , Chernin L.
Published: July 2010

This study showed that various rhizosphere bacteria producing the enzyme 1‐aminocyclopropane‐1‐carboxylate (ACC) deaminase (ACCD), which can degrade ACC, the immediate precursor of ethylene in plants, and thereby lower plant ethylene levels, can act as promising biocontrol agents of pathogenic strains of Agrobacterium tumefaciens and A. vitis. Soaking the roots of tomato (Solanum lycopersicum) seedlings in a suspension of the ACCD‐producing Pseudomonas putida UW4, Burkholderia phytofirmans PsJN or Azospirillum brasilense Cd1843 transformed by plasmid pRKTACC carrying the ACCD‐encoding gene acdS from UW4, significantly reduced the development of tumours on tomato plants injected 4–5 days later with pathogenic Agrobacterium strains via wounds on the plant stem. The fresh mass of tumours formed by plants pretreated with ACCD‐producing strains was typically four‐ to fivefold less than that of tumours formed on control plants inoculated only with a pathogenic Agrobacterium strain. Simultaneously, the level of ethylene evolution per amount of tumour mass on plants pretreated with ACCD‐producing bacteria decreased four to eight times compared with that from tumours formed on control plants or plants pretreated with bacteria deficient in ACCD production. Moreover, transgenic tomato plants expressing a bacterial ACCD were found to be highly resistant to crown gall formation relative to the parental, non‐transformed tomato plants. The results support the hypothesis that ethylene is a crucial factor in Agrobacterium tumour formation, and that ACCD‐produced rhizosphere bacteria may protect plants infected by pathogenic Agrobacteria from crown gall disease.
Alopecia areata in Eringer cows
Authors: Timm Katrin , Rüfenacht Silvia , Von Tscharner Claudia , Bornand Valérie F. , Doherr Marcus G. , Oevermann Anna , Flury Christine , Rieder Stefan , Hirsbrunner Gaby , Drögemüller Cord , Roosje Petra J.
Published: July 2010

Alopecia areata is a hair loss disorder in humans, dogs and horses with a suspected autoimmune aetiology targeting anagen hair follicles. Alopecia areata is only sporadically reported in cows. Recently, we observed several cases of suspected alopecia areata in Eringer cows. The aim of this study was to confirm the presumptive diagnosis of alopecia areata and to define the clinical phenotype and histopathological patterns, including characterization of the infiltrating inflammatory cells. Twenty Eringer cows with alopecia and 11 Eringer cows without skin problems were included in this study. Affected cows had either generalized or multifocal alopecia or hypotrichosis. The tail, forehead and distal extremities were usually spared. Punch biopsies were obtained from the centre and margin of alopecic lesions and normal haired skin. Histological examination revealed several alterations in anagen hair bulbs. These included peri‐ and intrabulbar lymphocytic infiltration, peribulbar fibrosis, degenerate matrix cells with clumped melanosomes and pigmentary incontinence. Mild lymphocytic infiltrative mural folliculitis was seen in the inferior segment and isthmus of the hair follicles. Hair shafts were often unpigmented and dysplastic. The large majority of infiltrating lymphocytes were CD3+ T cells, whereas only occasional CD20+ lymphocytes were present in the peribulbar infiltrate. Our findings confirm the diagnosis of T‐cell‐mediated alopecia areata in these cows. Alopecia areata appears to occur with increased frequency in the Eringer breed, but distinct predisposing factors could not be identified.
Maze Learning and Recall in a Weakly Electric Fish, Mormyrus rume proboscirostris Boulenger (Mormyridae, Teleostei)1
Authors: Walton Alice G. , Moller Peter
Published: July 2010

Mormyrus rume proboscirostris, African weakly electric fish, were trained to seek shelter in a meander maze, and following path acquisition released into the empty arena with all maze cues removed, either from the original start box or from a novel site (recall). We demonstrate that fish use their active electrosense, sight, and lateral line synergistically in maze acquisition and recall. In the presence of an electric roadmap consisting of an array of aluminum and Plexiglas objects, fish employed landmark orientation. But fish ignored visual markers and relied on internalized motor routines, which was inconsistent with evidence for cognitive mapping.