bluuebunny/crossref_metadata_2025 huggingface dataset:
Interleukin‐15 Induces Interleukin‐17 Production by Synovial T Cell Lines from Patients with Rheumatoid Arthritis
Authors: Halvorsen E. H. , Strønen E. , Hammer H. B. , Goll G. L. , Sollid L. M. , Molberg Ø.
Published: November 2010

IL‐17‐producing T cells (Th17 cells) are believed to contribute to local inflammation and joint damage in rheumatoid arthritis (RA). Limited data exist on Th17 cells located within the inflamed synovial tissue (ST) of patients with RA. Here, we aimed to generate polyclonal T cell lines (TCLs) from the RA ST and assess their cytokine production, including the effects of exogenous IL‐15 on IL‐17 production in vitro. For five patients with RA, polyclonal TCLs were established from ST obtained by joint surgery. Synovial TCLs were expanded and stimulated by anti‐CD3/CD28 microbeads and exogenous cytokines. Cytokine production was assessed by culture supernatant analyses and intracellular flow cytometry, and TCLs were sorted based on their surface expression of CCR6. In addition to IL‐17, we detected IL‐6, IL‐10, IFN‐γ and TNF‐α in the synovial TCL culture supernatants. Exogenous IL‐15 increased the production of IL‐17 as well as the other cytokines except IFN‐γ. For IL‐17, this effect was more pronounced after prolonged culture times. Intracellular flow cytometry confirmed the presence of IL‐17+ and IL‐17+IFN‐γ+ CD4+ T cells in the TCLs. IL‐17+ and IL‐17+IFN‐γ+ T cells were enriched in the CD4+CCR6+ population. In conclusion, Th17 cells can be detected after polyclonal expansion and stimulation of RA synovial TCLs generated by joint surgery. The Th17 cells from the RA ST were enriched in the CD4+CCR6+ population, and they were sensitive to exogenous IL‐15. Th17 cells present within the synovial compartment may contribute to the RA pathogenesis and local joint damage.
Modulation of sugar metabolism by an INDETERMINATE DOMAIN transcription factor contributes to photoperiodic flowering in Arabidopsis
Authors: Seo Pil Joon , Ryu Jaeyong , Kang Seok Ki , Park Chung‐Mo
Published: November 2010

There has been a long‐standing interest in the role played by sugars in flowering. Of particular interest is how sugar‐related signals are integrated into flowering genetic pathways. Here, we demonstrate that the INDETERMINATE DOMAIN transcription factor AtIDD8 regulates photoperiodic flowering by modulating sugar transport and metabolism. We found that whereas AtIDD8‐deficient idd8 mutants exhibit delayed flowering under long days, AtIDD8‐overexpressing plants (35S:IDD8) show early flowering. In addition, the sucrose synthase genes SUS1 and SUS4 were upregulated in 35S:IDD8 plants but downregulated in idd8 mutants, in which endogenous sugar levels were altered. AtIDD8 activates the SUS4 gene by binding directly to its promoter, resulting in promoted flowering in SUS4‐overexpressing plants. SUS4 expression also responds to photoperiodic signals. Notably, the AtIDD8 gene is suppressed by sugar deprivation. Therefore, we conclude that AtIDD8 regulation of sugar transport and metabolism is linked to photoperiodic flowering.
House dust and storage mite contamination of dry dog food stored in open bags and sealed boxes in 10 domestic households
Authors: Gill Christina , McEwan Neil , McGarry John , Nuttall Tim
Published: November 2010

Dry pet food is a potential source of exposure to house dust and storage mite allergens in canine atopic dermatitis. This study evaluated contamination of house dust and dry dog food stored in paper bags, sealable plastic bags and sealable plastic boxes in 10 households for 90 days using Acarex® tests for guanine, a Der p 1 ELISA and mite flotation. Acarex® tests were negative in all the food samples but positive in all the house dust samples. The Der p 1 levels and mite numbers significantly increased in food from paper bags (P = 0.0073 and P = 0.02, respectively), but not plastic bags or boxes. Mite numbers and Der p 1 levels were 10–1000 times higher in house dust than the corresponding food samples (P < 0.0001). There were significant correlations between Der p 1 in house dust and food from the paper (P < 0.0001) and plastic bags (P = 0.003), and mite numbers in house dust and food from the paper bags (P = 0.0007). Bedding and carpets were significantly associated with Der p 1 levels in house dust (P = 0.015 and P = 0.01, respectively), and food from the paper (both P = 0.02) and plastic bags (P = 0.03 and P = 0.04, respectively). Mites were identified in six of 10 paper bag, three of 10 plastic bag, one of 10 plastic box and nine of 10 house dust samples. These comprised Dermatophagoides (54%), Tyrophagus (10%; all from food) and unidentified mites (36%). Storage of food in sealable plastic boxes largely prevented contamination for 3 months. Exposure to mites and mite proteins in all the stored food, however, appeared to be trivial compared with house dust.
Cells dispersed from Marinobacter hydrocarbonoclasticus SP17 biofilm exhibit a specific protein profile associated with a higher ability to reinitiate biofilm development at the hexadecane–water interface
Authors: Vaysse Pierre‐Joseph , Sivadon Pierre , Goulas Philippe , Grimaud Régis
Published: November 2010

Biofilm formation by marine hydrocarbonoclastic bacteria is commonly observed and has been recognized as an important mechanism for the biodegradation of hydrocarbons. In order to colonize new oil‐water interfaces, surface‐attached communities of hydrocarbonoclastic bacteria must release cells into the environment. Here we explored the physiology of cells freshly dispersed from a biofilm of Marinobacter hydrocarbonoclasticus developing at the hexadecane–water interface, by combining proteomic and physiological approaches. The comparison of the dispersed cells' proteome with those of biofilm, logarithmic‐ and stationary‐phase planktonic cells indicated that dispersed cells had lost most of the biofilm phenotype and expressed a specific proteome. Two proteins involved in cell envelope maturation, DsbA and CtpA, were exclusively detected in dispersed cells, suggesting a reshaping of the cell envelopes during biofilm dispersal. Furthermore, dispersed cells exhibited a higher affinity for hexadecane and initiated more rapidly biofilm formation on hexadecane than the reference planktonic cells. Interestingly, storage wax esters were rapidly degraded in dispersed cells, suggesting that their observed physiological properties may rely on reserve mobilization. Thus, by promoting oil surface colonization, cells emigrating from the biofilm could contribute to the success of marine hydrocarbonoclastic bacteria in polluted environments.
Children's Schooling and Parental Migration: Empirical Evidence on the ‘Left‐behind’ Generation in Albania
Authors: Giannelli Gianna Claudia , Mangiavacchi Lucia
Published: November 2010

This article investigates the long‐term effects of parental migration abroad on the schooling of children left behind in Albania. Although parents' migration usually benefits children economically, the lack of parental care may cause relational and psychological problems that may affect children's welfare in the long term. The phenomenon of children left behind — mainly by fathers — is considerable in Albania, where migration has represented the only viable way to cope with an increasing poverty and the scarcity of public resources for sustaining households' incomes. Between 1990 and 2005 in Albania 22 per cent of children under 18 have been left behind, with an average parental absence of 9 months. Using detailed information on family migration drawn from the Living Standard Measurement Survey for 2005, multiple‐choice models are applied to evaluate the school progression of older children and adolescents. A duration analysis of school participation with both discrete and continuous time models is then performed. The results show that past parental migration has a negative effect on school attendance in the long term with higher hazards of school dropouts for children left behind. These results are robust to the use of different econometric techniques and model specifications.
Iron handling in hippocampal neurons: activity‐dependent iron entry and mitochondria‐mediated neurotoxicity
Authors: Pelizzoni Ilaria , Macco Romina , Morini Marco Francesco , Zacchetti Daniele , Grohovaz Fabio , Codazzi Franca
Published: November 2010

The characterization of iron handling in neurons is still lacking, with contradictory and incomplete results. In particular, the relevance of non‐transferrin‐bound iron (NTBI), under physiologic conditions, during aging and in neurodegenerative disorders, is undetermined. This study investigates the mechanisms underlying NTBI entry into primary hippocampal neurons and evaluates the consequence of iron elevation on neuronal viability. Fluorescence‐based single cell analysis revealed that an increase in extracellular free Fe2+ (the main component of NTBI pool) is sufficient to promote Fe2+ entry and that activation of either N‐methyl‐d‐aspartate receptors (NMDARs) or voltage operated calcium channels (VOCCs) significantly potentiates this pathway, independently of changes in intracellular Ca2+ concentration ([Ca2+]i). The enhancement of Fe2+ influx was accompanied by a corresponding elevation of reactive oxygen species (ROS) production and higher susceptibility of neurons to death. Interestingly, iron vulnerability increased in aged cultures. Scavenging of mitochondrial ROS was the most powerful protective treatment against iron overload, being able to preserve the mitochondrial membrane potential and to safeguard the morphologic integrity of these organelles. Overall, we demonstrate for the first time that Fe2+ and Ca2+ compete for common routes (i.e. NMDARs and different types of VOCCs) to enter primary neurons. These iron entry pathways are not controlled by the intracellular iron level and can be harmful for neurons during aging and in conditions of elevated NTBI levels. Finally, our data draw the attention to mitochondria as a potential target for the treatment of the neurodegenerative processes induced by iron dysmetabolism.
Stereo‐selective inhibition of transient receptor potential TRPC5 cation channels by neuroactive steroids
Authors: Majeed Y , Amer MS , Agarwal AK , McKeown L , Porter KE , O'Regan DJ , Naylor J , Fishwick CWG , Muraki K , Beech DJ
Published: November 2010

BACKGROUND AND PURPOSE Transient receptor potential canonical 5 (TRPC5) channels are widely expressed, including in the CNS, where they potentiate fear responses. They also contribute to other non‐selective cation channels that are stimulated by G‐protein‐coupled receptor agonists and lipid and redox factors. Steroids are known to modulate fear and anxiety states, and we therefore investigated whether TRPC5 exhibited sensitivity to steroids.

EXPERIMENTAL APPROACH Human TRPC5 channels were conditionally expressed in HEK293 cells and studied using intracellular Ca2+ measurement, whole‐cell voltage‐clamp and excised patch techniques. For comparison, control experiments were performed with cells lacking TRPC5 channels or expressing another TRP channel, TRPM2. Native TRPC channel activity was recorded from vascular smooth muscle cells.

KEY RESULTS Extracellular application of pregnenolone sulphate, pregnanolone sulphate, pregnanolone, progesterone or dihydrotestosterone inhibited TRPC5 activity within 1–2 min. Dehydroepiandrosterone sulphate or 17β‐oestradiol had weak inhibitory effects. Pregnenolone, and allopregnanolone, a progesterone metabolite and stereo‐isomer of pregnanolone, all had no effects. Progesterone was the most potent of the steroids, especially against TRPC5 channel activity evoked by sphingosine‐1‐phosphate. In outside‐out patch recordings, bath‐applied progesterone and dihydrotestosterone had strong and reversible effects, suggesting relatively direct mechanisms of action. Progesterone inhibited native TRPC5‐containing channel activity, evoked by oxidized phospholipid.

CONCLUSIONS AND IMPLICATIONS Our data suggest that TRPC5 channels are susceptible to relatively direct and rapid stereo‐selective steroid modulation, leading to channel inhibition. The study adds to growing appreciation of TRP channels as non‐genomic steroid sensors.
Sustainable transport: A case study of Rhine navigation
Authors: Pauli Gernot
Published: November 2010

In the debate about the future of the world's economies and in particular its transport systems, the concept of “sustainability” plays a prominent role. In this context, inland navigation is often cited as the most environmentally friendly and sustainable mode of transport, with fewer harmful emissions and less consumption of resources than road or rail transport. Using navigation in the Rhine basin as an example, this article sets out to: prove that inland navigation is becoming more sustainable; explain how that has happened; and identify the most important factors determining the sustainability of inland navigation. The article concludes that a new approach to the development of transport policy is needed and that this approach has to ensure important factors determining sustainability are integrated within a single policy.
Making climate instruments work for sustainable transport in developing countries
Authors: Bakker Stefan , Huizenga Cornie
Published: November 2010

In order to meet long‐term climate change mitigation objectives, emissions cuts are required in all regions across the globe and in all sectors, including transport. In financing this effort, the Clean Development Mechanism (CDM) and the Global Environmental Facility (GEF) are until now the only international climate policy instruments under the United Nations Framework Convention on Climate Change that provide incentives for emissions reductions in developing countries. More recently, the Clean Technology Fund (CTF) was established. In this paper, we show that the impact of these financing instruments on transport has been very limited, due to methodological difficulties, a data‐intensive monitoring process and the limited funding available. We argue that the transport sector is not likely to play a significant role in the continuation of a carbon credit offsetting scheme, unless these methodological requirements are simplified and significantly more funding is available.

In the post‐2012 climate regime, there may be substantial international funding available in addition to existing credit schemes and international funds, which could be channeled through nationally appropriate mitigation actions (NAMAs). This can provide new and better opportunities for sustainable transport in developing countries. We propose a framework for NAMAs, including types of policies and measures, measurement, reporting and verification of the actions, and an institutional and financial structure. We conclude that climate funding needs to be aligned closely with domestic and multilateral development finance flows in order to make a difference for sustainable transport.
Employment protection legislation and job stability: A European cross‐country analysis
Authors: CAZES Sandrine , TONIN Mirco
Published: November 2010

Based on Eurostat data on wage employment for the period 1999–2006, this article investigates the dynamics of job tenure across the European Union. The authors’ analysis shows no generalized decline in job tenure, but a trend towards shorter tenure among young workers (aged 15–24 years) in many European countries. Their regression results indicate that this trend is associated with the weakening of employment protection provided by law and by trade unions. Given young workers’ weak individual bargaining power, the trend towards individualization of the employment relationship may thus affect them disproportionately.
Female executives and the glass ceiling in Spain
Authors: Castaño Cecilia , Martín Juan , Vázquez Susana , Martínez José Luís
Published: November 2010

This paper examines the findings of a survey of the personal, educational and professional profiles of a sample of male and female senior executives of companies operating in Spain. The women were found to be younger; they had fewer children, resorted to more domestic help, and earned less than the men, although they often had higher qualifications. Yet cultural patterns and institutional barriers still prevent them from making full use of their capabilities. In spite of recent legislative efforts to promote gender equity at work, more than nine out of ten senior executives are still men.
Thailand's work and health transition
Authors: Kelly Matthew , Strazdins Lyndall , Ellora Tarie D , Khamman Suwanee , Seubsman Sam‐ang , Sleigh Adrian C.
Published: November 2010

Thailand has experienced a rapid economic transition from agriculture to manufacturing and services, and to more formal employment. Its labour market regulation and worker representation, however, are much weaker than they are in developed countries, which underwent these transitions more slowly and sequentially, decades earlier. The authors examine the strengthening of Thailand's policy and legislation on occupational safety and health in response to international standards, a new democratic Constitution, fear of foreign trade embargoes, and fatal workplace disasters. In concluding, they identify key challenges remaining for policy‐makers, including enforcement of legislation and measurement of new mental and physical health effects.
The Role of Ubiquitination in Lysosomal Trafficking of δ‐Opioid Receptors
Authors: Henry Anastasia G. , White Ian J. , Marsh Mark , von Zastrow Mark , Hislop James N.
Published: November 2010

The δ‐opioid receptor (DOR) undergoes ligand‐induced downregulation by endosomal sorting complex required for transport (ESCRT)‐dependent endocytic trafficking to lysosomes. In contrast to a number of other signaling receptors, the DOR can downregulate effectively when its ubiquitination is prevented. We explored the membrane trafficking basis of this behavior. First, we show that internalized DORs traverse the canonical multivesicular body (MVB) pathway and localize to intralumenal vesicles (ILVs). Second, we show that DOR ubiquitination stimulates, but is not essential for, receptor transfer to ILVs and proteolysis of the receptor endodomain. Third, we show that receptor ubiquitination plays no detectable role in the early sorting of internalized DORs out of the recycling pathway. Finally, we show that DORs undergo extensive proteolytic fragmentation in the ectodomain, even when receptor ubiquitination is prevented or ILV formation itself is blocked. Together, these results are sufficient to explain why DORs downregulate effectively in the absence of ubiquitination, and they place a discrete molecular sorting operation in the MVB pathway effectively upstream of the ESCRT. More generally, these findings support the hypothesis that mammalian cells can control the cytoplasmic accessibility of internalized signaling receptors independently from their ultimate trafficking fate.
Nursing assessment during oxygen administration in ventilated preterm infants
Authors: Solberg Marianne T , Hansen Thor WR , Bjørk Ida Torunn
Published: November 2010

Aim:  To document nurses’ opinions about their assessments of oxygen requirements in ventilated preterm infants receiving oxygen supplementation.

Methods:  Survey design with descriptive statistics. The sample consisted of 111 nurses employed in clinical positions in neonatal intensive care units within Norway’s five regional hospitals. The questionnaire included questions about physiological and clinical observations used when assessing oxygen administration in ventilated preterm infants.

Results:  A major finding was the gap between the criteria laid down in professional and research literature, vs the criteria nurses perceived they were using when they assessed the oxygen requirements of preterm infants. The respondents stated that they used oxygen saturation to assess the infants’ oxygen requirements when adjusting oxygen supplementation. Only 17% of the nurses used the oxygen‐haemoglobin dissociation curve in their assessments. Those who responded that they used the curve did not use it correctly.

Conclusion:  Assessment of O2 requirement is based on insufficient information and calls for evaluation of local, national as well as international education. Close collaboration between doctors and nurses is essential in planning care for individual patients.
Ion channel activity of brain abundant protein BASP1 in planar lipid bilayers
Authors: Ostroumova Olga S. , Schagina Ludmila V. , Mosevitsky Mark I. , Zakharov Vladislav V.
Published: November 2010

BASP1 (also known as CAP‐23 and NAP‐22) is a brain abundant myristoylated protein localized at the inner surface of the presynaptic plasma membrane. Emerging evidence suggests that BASP1 is critically involved in various cellular processes, in particular, in the accumulation of phosphatidylinositol‐4,5‐diphosphate (PIP2) in lipid raft microdomains. We have recently shown that BASP1 forms heterogeneously‐sized oligomers and higher aggregates with an outward similarity to oligomers and protofibrils of amyloid proteins. However, BASP1 is not known to be related to any amyloid disease. In the present study, we show that BASP1 induces single channel currents across negatively‐charged planar lipid bilayers (containing phosphatidylserine or PIP2) bathed in 0.1–0.2 m KCl (pH 7.5). By their characteristics, BASP1 channels are similar to amyloid protein channels. BASP1 channels exhibit multiple conductance levels, in the range 10–3000 pS, with the most frequently observed conductance state of approximately 50 pS. The channels demonstrate a linear current–voltage relationship and voltage‐independent kinetics of opening and closing. Their K+ to Cl− permeability ratio is approximately 14, indicating that BASP1 channels are cation‐selective. The ion channel activity of BASP1 is in accordance with the pore‐like structure of BASP1 oligomers observed by electron microscopy on a lipid monolayer. Neuronal protein GAP‐43, which is functionally related to BASP1 and also forms oligomers, elicited no ion channel currents under the conditions used in the present study. Elucidation of the physiological or pathological roles of ion channel activity of membrane‐bound BASP1 oligomers will help to define the precise mechanism of amyloid protein toxicity.
The mitochondrial protein frataxin is essential for heme biosynthesis in plants
Authors: Maliandi María V. , Busi Maria V. , Turowski Valeria R. , Leaden Laura , Araya Alejandro , Gomez‐Casati Diego F.
Published: November 2010

Frataxin, a conserved mitochondrial protein implicated in cellular iron homeostasis, has been involved as the iron chaperone that delivers iron for the Fe–S cluster and heme biosynthesis. However, its role in iron metabolism remains unclear, especially in photosynthetic organisms. In previous work, we found that frataxin deficiency in Arabidopsis results in decreased activity of the mitochondrial Fe–S proteins aconitase and succinate dehydrogenase, despite the increased expression of the respective genes, indicating an important role for Arabidopsis thaliana frataxin homolog (AtFH). In this work, we explore the hypothesis that AtFH can participate in heme formation in plants. For this purpose, we used two Arabidopsis lines, atfh‐1 and as‐AtFH, with deficiency in the expression of AtFH. Both lines present alteration in several transcripts from the heme biosynthetic route with a decrease in total heme content and a deficiency in catalase activity that was rescued with the addition of exogenous hemin. Our data substantiate the hypothesis that AtFH, apart from its role in protecting bioavailable iron within mitochondria and the biogenesis of Fe–S groups, also plays a role in the biosynthesis of heme groups in plants.
International and Regional Cooperation: Asia's Role and Responsibilities
Authors: DRYSDALE Peter , ARMSTRONG Shiro
Published: November 2010

Asia has emerged from the global financial crisis as an important stabilizing force and an engine of global economic growth. The establishment of the G‐20 gives Asian economies the global forum that they need to both represent their interests in global governance and deliver on responsibilities concomitant with their growing weight in the global economy. The region has a host of cooperation arrangements in APEC, ASEAN+3, and EAS (East Asian Summit), all with ASEAN as the fulcrum. They are huge assets, but they need to be repositioned to relate effectively to the G‐20 process and other global arrangements. They also need to comprehend the politics of the changing structure of regional power. This paper discusses the challenges that Asia faces in aligning regional and global objectives in financial, trade, and other areas of cooperation, such as climate change and foreign investment. It argues that Asia is now a critical player in the global system and has a central contribution to make in strengthening global governance and international policy outcomes. The paper sets out ways to fill gaps in regional cooperation and link the agenda for regional cooperation more effectively to Asia's new role globally. This is essential to sustain Asia's superior growth performance, correct imbalances, and support the global economic system.
Simple, fast and high‐efficiency transformation system for directed evolution of cellulase in Bacillus subtilis
Authors: Zhang Xiao‐Zhou , Zhang Y.‐H. Percival
Published: November 2010

Bacillus subtilis can serve as a powerful platform for directed evolution, especially for secretory enzymes. However, cloning and transformation of a DNA mutant library in B. subtilis are not as easy as they are in Escherichia coli. For direct transformation of B. subtilis, here we developed a new protocol based on supercompetent cells prepared from the recombinant B. subtilis strain SCK6 and multimeric plasmids. This new protocol is simple (restriction enzyme‐, phosphatase‐ and ligase‐free), fast (i.e. 1 day) and of high efficiency (i.e. ∼107 or ∼104 transformants per µg of multimeric plasmid or ligated plasmid DNA respectively). Supercompetent B. subtilis SCK6 cells were prepared by overexpression of the competence master regulator ComK that was induced by adding xylose. The DNA mutant library was generated through a two‐round PCR: (i) the mutagenized DNA fragments were generated by error‐prone PCR and linearized plasmids were made using high‐fidelity PCR, and (ii) the multimeric plasmids were generated based on these two DNA templates by using overlap PCR. Both protein expression level and specific activity of glycoside hydrolase family 5 endoglucanse on regenerated amorphous cellulose were improved through this new system. To our limited knowledge, this study is the first report for enhancing secretory cellulase performance on insoluble cellulose.
Adherence with brimonidine in patients with glaucoma aware and not aware of electronic monitoring
Authors: Hermann Manuel Marcel , Papaconstantinou Dimitris , Muether Philipp Sebastian , Georgopoulos Gerasimos , Diestelhorst Michael
Published: November 2010

Purpose:  To assess the impact of open versus masked adherence monitoring on adherence with topical brimonidine using two different dosing schedules.

Methods:  Thirty‐seven patients with glaucoma or ocular hypertension were enroled in a prospective, observational cohort study. Patients were randomly assigned to open or masked adherence monitoring and to brimonidine twice daily (BID) or three times daily (TID). Patients received conventional brimonidine eye drops with attached electronic monitoring devices for 4 weeks with weekly intraocular pressure measurements. Adherence calculations comprised dosing intervals, adherence rate and time covered. Subgroup analysis with anova included the factors masking, regimen, diagnosis and age.

Results:  Among 36 individually analysed patients, 12 (33%) had adherence rates above 75%, therein two (5%)>90%. The mean adherence rate in 19 patients aware of adherence measurements was 70 ± 17% for brimonidine BID and 65 ± 14% for TID, not significantly different to the rates of 17 patients not informed about adherence measurements (77 ± 6% BID, 62 ± 9% TID, p = 0.24). On average, patients with brimonidine TID achieved significantly lower adherence rates (64 ± 12%) than patients on BID (73 ± 13%, p = 0.02). Still, patients on TID applied brimonidine more often (TID 1.9 ± 0.3, BID 1.5 ± 0.1 mean applications per day). The median coverage was 70% and showed no statistically significant difference between patients on BID and TID (p = 0.36).

Conclusion:  The study findings suggest that adherence measurements are not significantly altered by open adherence monitoring, which may simplify future adherence studies. Adherence with brimonidine eye drops was insufficient for most patients. These results demonstrate the necessity to develop new strategies to improve compliance in glaucoma therapy.
Real orthopaedic subluxations versus imaginary chiropractic subluxations
Authors: Homola Samuel
Published: November 2010

The chiropractic vertebral subluxation theory has confused the definition of the word ‘subluxation’; a common medical term. Unlike the mysterious, undetectable chiropractic ‘vertebral subluxation complex’ alleged to be a cause of disease, a real vertebral subluxation can be a cause of mechanical and neuromusculoskeletal symptoms, but has never been shown to be a cause of organic disease. The views offered in this paper may be helpful in clarifying this issue.
The Core Structure of Basal Dislocations in Deformed Sapphire (α-Al
2
O
3
)
Authors: Heuer A. H. , Jia C. L. , Lagerlöf K. P. D.
Published: November 2010

Deformed Sapphire

The structure of sapphire consists of alternating sheets of oxygen ions and puckered aluminum ions. Although brittle at low temperature, the material can deform at high temperatures through structural dislocations.

Heuer
et al.

(p.
1227
) used a high-resolution transmission electron microscope and the negative spherical aberration imaging technique to distinguish between the columns of Al and O ions. Upon deformation, the Al ions moved into both tetrahedrally and octahedrally coordinated sites, and while the dislocation cores spread out, they remained uncharged and stoichiometric. The results may connect to the deformation behavior of other nonmetallic solids.
From the Connectome to the Synaptome: An Epic Love Story
Authors: DeFelipe Javier
Published: November 2010

Connections by Degree

A recent explosion of data and of large projects dealing with the problem of connectivity in the human brain has not yet led to clear strategies to study connectomes.
DeFelipe
(p.
1198
) reviews the intellectual history of the field, from the 19th-century studies of the seminal anatomist Ramon y Cajal to the very recent approaches and myriad new technologies applied by many laboratories trying to understand the organization of the human cerebral cortex.
Reducing the Gender Achievement Gap in College Science: A Classroom Study of Values Affirmation
Authors: Miyake Akira , Kost-Smith Lauren E. , Finkelstein Noah D. , Pollock Steven J. , Cohen Geoffrey L. , Ito Tiffany A.
Published: November 2010

Writing to Close Gaps

Some have questioned whether findings in the laboratory obtained under controlled conditions and limited contexts bear any relevance to behavior in real-world environments in which ordinary people cope with real-life challenges. Recent studies have shown a replicable and long-term effect of a brief writing exercise on the academic performance of African-American seventh graders in an inner-city public school.

Miyake
et al.

(p.
1234
) extended this approach to show that a similar kind of writing exercise can help to reduce the gender gap observed in the performance of female students in an undergraduate physics class, where performance is measured not only via course grades and exam scores, but also on a standardized test.
Intermediate Steps
Authors: Devos Damien P. , Reynaud Emmanuel G.
Published: November 2010

Bacteria that also have features typical of eukaryotes and archaea may reflect a possible pathway in ancient cellular evolution.
The Energy-Poverty-Climate Nexus
Authors: Casillas Christian E. , Kammen Daniel M.
Published: November 2010

Community-level carbon abatement curves highlight opportunities for increased access to clean, efficient energy for the poor.
Complete Biodegradation of 4-Fluorocinnamic Acid by a Consortium Comprising
Arthrobacter
sp. Strain G1 and
Ralstonia
sp. Strain H1
Authors: Hasan Syed A. , Ferreira Maria Isabel M. , Koetsier Martijn J. , Arif Muhammad I. , Janssen Dick B.
Published: November 2010

A consortium of the newly isolated bacterial strains
Arthrobacter
sp. strain G1 and
Ralstonia
sp. strain H1 utilized 4-fluorocinnamic acid for growth under aerobic conditions. Strain G1 converted 4-fluorocinnamic acid into 4-fluorobenzoic acid and used the two-carbon side chain for growth, with some formation of 4-fluoroacetophenone as a dead-end side product. In the presence of strain H1, complete mineralization of 4-fluorocinnamic acid and release of fluoride were obtained. Degradation of 4-fluorocinnamic acid by strain G1 occurred through a β-oxidation mechanism and started with the formation of 4-fluorocinnamoyl-coenzyme A (CoA), as indicated by the presence of 4-fluorocinnamoyl-CoA ligase. Enzymes for further transformation were detected in cell extract, i.e., 4-fluorocinnamoyl-CoA hydratase, 4-fluorophenyl-β-hydroxy propionyl-CoA dehydrogenase, and 4-fluorophenyl-β-keto propionyl-CoA thiolase. Degradation of 4-fluorobenzoic acid by strain H1 proceeded via 4-fluorocatechol, which was converted by an
ortho
-cleavage pathway.
Mycolic Acid-Containing Bacteria Induce Natural-Product Biosynthesis in
Streptomyces
Species
Authors: Onaka Hiroyasu , Mori Yukiko , Igarashi Yasuhiro , Furumai Tamotsu
Published: November 2010

Natural products produced by microorganisms are important starting compounds for drug discovery. Secondary metabolites, including antibiotics, have been isolated from different
Streptomyces
species. The production of these metabolites depends on the culture conditions. Therefore, the development of a new culture method can facilitate the discovery of new natural products. Here, we show that mycolic acid-containing bacteria can influence the biosynthesis of cryptic natural products in
Streptomyces
species. The production of red pigment by
Streptomyces lividans
TK23 was induced by coculture with
Tsukamurella pulmonis
TP-B0596, which is a mycolic acid-containing bacterium. Only living cells induced this pigment production, which was not mediated by any substances.
T
.
pulmonis
could induce natural-product synthesis in other
Streptomyces
strains too: it altered natural-product biosynthesis in 88.4% of the
Streptomyces
strains isolated from soil. The other mycolic acid-containing bacteria,
Rhodococcus erythropolis
and
Corynebacterium glutamicum
, altered biosynthesis in 87.5 and 90.2% of the
Streptomyces
strains, respectively. The coculture broth of
T
.
pulmonis
and
Streptomyces endus
S-522 contained a novel antibiotic, which we named alchivemycin A. We concluded that the mycolic acid localized in the outer cell layer of the inducer bacterium influences secondary metabolism in
Streptomyces
, and this activity is a result of the direct interaction between the mycolic acid-containing bacteria and
Streptomyces
. We used these results to develop a new coculture method, called the combined-culture method, which facilitates the screening of natural products.
Construction and Characterization of a Lactose-Inducible Promoter System for Controlled Gene Expression in
Clostridium perfringens
Authors: Hartman Andrea H. , Liu Hualan , Melville Stephen B.
Published: November 2010

Clostridium perfringens
is a Gram-positive anaerobic pathogen which causes many diseases in humans and animals. While some genetic tools exist for working with
C. perfringens
, a tightly regulated, inducible promoter system is currently lacking. Therefore, we constructed a plasmid-based promoter system that provided regulated expression when lactose was added. This plasmid (pKRAH1) is an
Escherichia coli-C. perfringens
shuttle vector containing the gene encoding a transcriptional regulator, BgaR, and a divergent promoter upstream of gene
bgaL
(
bgaR
-P

bgaL

). To measure transcription at the
bgaL
promoter in pKRAH1, the
E. coli
reporter gene
gusA
, encoding β-glucuronidase, was placed downstream of the P

bgaL

promoter to make plasmid pAH2. When transformed into three strains of
C. perfringens
, pAH2 exhibited lactose-inducible expression.
C. perfringens
strain 13, a commonly studied strain, has endogenous β-glucuronidase activity. We mutated gene
bglR
, encoding a putative β-glucuronidase, and observed an 89% decrease in endogenous activity with no lactose. This combination of a system for regulated gene expression and a mutant of strain 13 with low β-glucuronidase activity are useful tools for studying gene regulation and protein expression in an important pathogenic bacterium. We used this system to express the
yfp
-
pilB
gene, comprised of a yellow fluorescent protein (YFP)-encoding gene fused to an assembly ATPase gene involved in type IV pilus-dependent gliding motility in
C. perfringens
. Expression in the wild-type strain showed that YFP-PilB localized mostly to the poles of cells, but in a
pilC
mutant it localized throughout the cell, demonstrating that the membrane protein PilC is required for polar localization of PilB.
Utilization of Natural Fucosylated Oligosaccharides by Three Novel α-
l
-Fucosidases from a Probiotic
Lactobacillus casei
Strain
Authors: Rodríguez-Díaz Jesús , Monedero Vicente , Yebra María J.
Published: November 2010

Three putative α-
l
-fucosidases encoded in the
Lactobacillus casei
BL23 genome were cloned and purified. The proteins displayed different abilities to hydrolyze natural fucosyloligosaccharides like 2′-fucosyllactose, H antigen disaccharide, H antigen type II trisaccharide, and 3′-, 4′-, and 6′-fucosyl-GlcNAc. This indicated a possible role in the utilization of oligosaccharides present in human milk and intestinal mucosa.
Acute Parvovirus B19 Infection Causes Nonspecificity Frequently in
Borrelia
and Less Often in
Salmonella
and
Campylobacter
Serology, Posing a Problem in Diagnosis of Infectious Arthropathy
Authors: Tuuminen Tamara , Hedman Klaus , Söderlund-Venermo Maria , Seppälä Ilkka
Published: November 2010

Several infectious agents may cause arthritis or arthropathy. For example, infection with
Borrelia burgdorferi
, the etiologic agent of Lyme disease, may in the late phase manifest as arthropathy. Infections with
Campylobacter
,
Salmonella
, or
Yersinia
may result in a postinfectious reactive arthritis. Acute infection with parvovirus B19 (B19V) may likewise initiate transient or chronic arthropathy. All these conditions may be clinically indistinguishable from rheumatoid arthritis. Here, we present evidence that acute B19V infection may elicit IgM antibodies that are polyspecific or cross-reactive with a variety of bacterial antigens. Their presence may lead to misdiagnosis and improper clinical management, exemplified here by two case descriptions. Further, among 33 subjects with proven recent B19V infection we found IgM enzyme immunoassay (EIA) positivity for
Borrelia
only; for
Borrelia
and
Salmonella
; for
Borrelia
and
Campylobacter
; and for
Borrelia
,
Campylobacter
, and
Salmonella
in 26 (78.7%), 1 (3%), 2 (6%), and 1 (3%), respectively; however, when examined by
Borrelia
LineBlot, all samples were negative. These antibodies persisted over 3 months in 4/13 (38%) patients tested. Likewise, in a retrospective comparison of the results of a diagnostic laboratory, 9/11 (82%) patients with confirmed acute B19V infection showed IgM antibody to
Borrelia
. However, none of 12 patients with confirmed borreliosis showed any serological evidence of acute B19V infection. Our study demonstrates that recent B19V infection can be misinterpreted as secondary borreliosis or enteropathogen-induced reactive arthritis. To obtain the correct diagnosis, we emphasize caution in interpretation of polyreactive IgM and exclusion of recent B19V infection in patients examined for infectious arthritis or arthropathy.
Differential Responses of
Bacillus subtilis
rRNA Promoters to Nutritional Stress
Authors: Samarrai Walied , Liu David X. , White Ann-Marie , Studamire Barbara , Edelstein Jacob , Srivastava Anita , Widom Russell L. , Rudner Rivka
Published: November 2010

The
in vivo
expression levels of four rRNA promoter pairs (
rrn
p
1
p
2
) of
Bacillus subtilis
were determined by employing single-copy
lacZ
fusions integrated at the
amyE
locus. The
rrnO
,
rrnJ
,
rrnD
, and
rrnB
promoters displayed unique growth rate regulation and stringent responses. Both
lacZ
activity and mRNA levels were highest for
rrnO
under all growth conditions tested, while
rrnJ
,
rrnB
,
and rrnD
showed decreasing levels of activity. During amino acid starvation induced by serine hydroxamate (SHX), only the strong
rrnO
and
rrnJ
promoters demonstrated stringent responses. Under the growth conditions used, the
rrn
promoters showed responses similar to the responses to carbon source limitation induced by α-methyl glucoside (α-MG). The ratio of P2 to P1 transcripts, determined by primer extension analysis, was high for the strong
rrnO
and
rrnJ
promoters, while only P2 transcripts were detected for the weak
rrnD
and
rrnB
promoters. Cloned P1 or P2 promoter fragments of
rrnO
or
rrnJ
were differentially regulated. In wild-type (

relA
+

) and suppressor [
relA
(S)] strains under the conditions tested, only P2 responded to carbon source limitation by a decrease in RNA synthesis, correlating with an increase in (p)ppGpp levels and a decrease in the GTP concentration. The weak P1 promoter elements remain relaxed in the three genetic backgrounds [

relA
+

,
relA
,
relA
(S)] in the presence of α-MG. During amino acid starvation, P2 was stringently regulated in

relA
+

and
relA
(S) cells, while only
rrnJ
p
1
was also regulated, but to a lesser extent. Both the
relA
+
and
relA
(S) strains showed (p)ppGpp accumulation after α-MG treatment but not after SHX treatment. These data reveal the complex nature of
B. subtilis rrn
promoter regulation in response to stress, and they suggest that the P2 promoters may play a more prominent role in the stringent response.
The Peptidoglycan-Binding Protein FimV Promotes Assembly of the
Pseudomonas aeruginosa
Type IV Pilus Secretin
Authors: Wehbi Hania , Portillo Eder , Harvey Hanjeong , Shimkoff Anthony E. , Scheurwater Edie M. , Howell P. Lynne , Burrows Lori L.
Published: November 2010

The
Pseudomonas aeruginosa
inner membrane protein FimV is among several proteins of unknown function required for type IV pilus-mediated twitching motility, arising from extension and retraction of pili from their site of assembly in the inner membrane. The pili transit the periplasm and peptidoglycan (PG) layer, ultimately exiting the cell through the PilQ secretin. Although
fimV
mutants are nonmotile, they are susceptible to killing by pilus-specific bacteriophage, a hallmark of retractable surface pili. Here we show that levels of recoverable surface pili were markedly decreased in
fimV pilT
retraction-deficient mutants compared with levels in the
pilT
control, demonstrating that FimV acts at the level of pilus assembly. Levels of inner membrane assembly subcomplex proteins PilM/N/O/P were decreased in
fimV
mutants, but supplementation of these components in
trans
did not restore pilus assembly or motility. Loss of FimV dramatically reduced the levels of the PilQ secretin multimer through which pili exit the cell, in part due to decreased levels of PilQ monomers, while PilF pilotin levels were unchanged. Expression of
pilQ
in
trans
in the wild type or
fimV
mutants increased total PilQ monomer levels but did not alter secretin multimer levels or motility. PG pulldown assays showed that the N terminus of FimV bound PG in a LysM motif-dependent manner, and a mutant with an in-frame chromosomal deletion of the LysM motif had reduced motility, secretin levels, and surface piliation. Together, our data show that FimV's role in pilus assembly is to promote secretin formation and that this function depends upon its PG-binding domain.
DNA-Binding Properties of the
Bacillus subtilis
and
Aeribacillus pallidus
AC6 σ
D
Proteins
Authors: Sevim Elif , Gaballa Ahmed , Beldüz A. Osman , Helmann John D.
Published: November 2010

σ
D
proteins from
Aeribacillus pallidus
AC6 and
Bacillus subtilis
bound specifically, albeit weakly, to promoter DNA even in the absence of core RNA polymerase. Binding required a conserved CG motif within the −10 element, and this motif is known to be recognized by σ region 2.4 and critical for promoter activity.
Enteric Campylobacteria and RNA Viruses Associated with Healthy and Diarrheic Humans in the Chinook Health Region of Southwestern Alberta, Canada
Authors: Inglis G. Douglas , Boras Valerie F. , Houde Alain
Published: November 2010

The presence of
Campylobacter
species and enteric RNA viruses in stools from diarrheic (
n
= 442) and healthy (
n
= 58) humans living in southwestern Alberta was examined (May to October 2005). A large number of diarrheic individuals who were culture negative for
C. jejuni
(
n
= 54) or
C. coli
(
n
= 19) were PCR positive for these taxa. Overall detection rates for
C. jejuni
and
C. coli
in diarrheic stools were 29% and 5%, respectively. In contrast, 3% and 0% of stools from healthy humans were positive for these taxa, respectively. Infection with
C. jejuni
was endemic over the study period. However, there was no difference in infection rates between individuals living in urban or rural locations. Stools from a large number of diarrheic (74%) and healthy (88%) individuals were positive for
Campylobacter
DNA. The prevalence rates of
C. concisus
,
C. curvus
,
C. fetus
,
C. gracilis
,
C. helveticus
,
C. hominis
,
C. hyointestinalis
,
C. mucosalis
,
C. showae
,
C. sputorum
, and
C. upsaliensis
DNA were either not significantly different or were significantly lower in stools from diarrheic than from healthy individuals. No
C. lanienae
or
C. lari
DNA was detected. Stools from 4% and 0% of diarrheic and healthy humans, respectively, were positive for rotavirus, sapovirus, or norovirus (GI/GII). Our results showed a high prevalence of diarrheic individuals living in southwestern Alberta who were infected by
C. jejuni
and, to a lesser extent, by
C. coli
. However, other
Campylobacter
species, norovirus, rotavirus, sapovirus, and bovine enteric calicivirus were either inconsequential pathogens during the study period or are not pathogens at all.