bluuebunny/crossref_metadata_2025 huggingface dataset:
Purification, crystallization and preliminary crystallographic analysis of human cystathionine β-synthase
Authors: Oyenarte Iker , Majtan Tomas , Ereño June , Corral-Rodríguez María Angeles , Kraus Jan P. , Martínez-Cruz Luis Alfonso
Published: October 2012

Human cystathionine β-synthase (CBS) is a pyridoxal-5′-phosphate-dependent hemeprotein, whose catalytic activity is regulated byS-adenosylmethionine. CBS catalyzes the β-replacement reaction of homocysteine (Hcy) with serine to yield cystathionine. CBS is a key regulator of plasma levels of the thrombogenic Hcy and deficiency in CBS is the single most common cause of homocystinuria, an inherited metabolic disorder of sulfur amino acids. The properties of CBS enzymes, such as domain organization, oligomerization degree or regulatory mechanisms, are not conserved across the eukaryotes. The current body of knowledge is insufficient to understand these differences and their impact on CBS function and physiology. To overcome this deficiency, we have addressed the crystallization and preliminary crystallographic analysis of a protein construct (hCBS516–525) that contains the full-length CBS fromHomo sapiens(hCBS) and just lacks amino-acid residues 516–525, which are located in a disordered loop. The human enzyme yielded crystals belonging to space groupI222, with unit-cell parametersa= 124.98,b= 136.33,c= 169.83 Å and diffracting X-rays to a resolution of 3.0 Å. The crystal structure appears to contain two molecules in the asymmetric unit which presumably correspond to a dimeric form of the enzyme.
Purification, crystallization and preliminary X-ray diffraction studies of the ATP-binding subunit of an ABC transporter fromGeobacillus kaustophilus
Authors: Manjula Mallappa , Pampa Kudigana J. , Madan Kumar Shankar , Kunishima Naoki , Lokanath Neratur K.
Published: October 2012

ATP-binding cassette (ABC) transporters, also known as traffic ATPases, form a large family of integral membrane proteins responsible for the translocation of a variety of chemically diverse substrates across the lipid bilayers of cellular membranes of both prokaryotes and eukaryotes by the hydrolysis of ATP. The ATP-binding subunit of an ABC transporter fromGeobacillus kaustophilus, a homodimeric enzyme, was overexpressed inEscherichia coliand purified. Crystals were obtained using the microbatch-under-oil method at 291 K. X-ray diffraction data to 1.6 Å resolution were collected on SPring-8 beamline BL26B1. The crystals belonged to the orthorhombic space groupI222, with unit-cell parametersa= 54.94,b = 78.63,c= 112.96 Å. Assuming the presence of a dimer in the asymmetric unit gave a crystal volume per protein weight (VM) of 2.32 Å3 Da−1and a solvent content of 47%; this was consistent with the results of a dynamic light-scattering experiment, which showed a dimeric state of the protein in solution. Molecular-replacement trials using the crystal structure of HisP from theSalmonella typhimuriumATP-binding subunit of an ABC transporter as a search model did not provide a satisfactory solution, indicating that the two ATP-binding subunits of ABC transporters have substantially different structures.
Purification, crystallization and preliminary X-ray diffraction analysis of an acidic phospholipase A2with vasoconstrictor activity fromAgkistrodon halys pallasvenom
Authors: Zou Zhisong , Zeng Fuxing , Zhang Lu , Niu Liwen , Teng Maikun , Li Xu
Published: October 2012

Phospholipases A2(PLA2s) are the major component of snake venoms and exert a variety of relevant toxic actions such as neurotoxicity and myotoxicity, amongst others. An acidic PLA2, here named AhV_aPA, was purified fromAgkistrodon halys pallasvenom by means of a three-step chromatographic procedure. AhV_aPA migrated as a single band on SDS–PAGE gels, with a molecular weight of about 14 kDa. Like other acidic aPLA2s, AhV_aPA has high enzymatic activity. Tension measurements of mouse thoracic aortic rings remarkably indicated that AhV_aPA could induce a further contractile response on the 60 mMK+-induced contraction, with an EC50of 369 nmol l−1. Rod-shaped crystals were obtained by the hanging-drop vapour-diffusion method and diffracted to a resolution limit of 2.30 Å. The crystals belonged to space groupP222, with unit-cell parametersa= 44.27,b= 68.39,c= 81.54 Å.
Purification, crystallization and preliminary crystallographic analysis of the full-length cystathionine β-synthase fromApis mellifera
Authors: Oyenarte Iker , Majtan Tomas , Ereño June , Corral-Rodríguez María Angeles , Klaudiny Jaroslav , Majtan Juraj , Kraus Jan P. , Martínez-Cruz Luis Alfonso
Published: October 2012

Cystathionine β-synthase (CBS) is a pyridoxal-5′-phosphate-dependent enzyme that catalyzes the first step of the transsulfuration pathway, namely the condensation of serine with homocysteine to form cystathionine. Mutations in the CBS gene are the single most common cause of hereditary homocystinuria, a multisystemic disease affecting to various extents the vasculature, connective tissues and central nervous system. At present, the crystal structure of CBS fromDrosophila melanogasteris the only available structure of the full-length enzyme. Here we describe a cloning, overexpression, purification and preliminary crystallographic analysis of a full-length CBS fromApis mellifera(AmCBS) which maintains 51 and 46% sequence identity with itsDrosophilaand human homologs, respectively. TheAmCBS yielded crystals belonging to space groupP212121, with unit-cell parametersa= 85.90,b= 95.87,c= 180.33 Å. Diffraction data were collected to a resolution of 3.0 Å. The crystal structure contained two molecules in the asymmetric unit which presumably correspond to the dimeric species observed in solution.
Crystallization and preliminary crystallographic analysis of 2-aminophenol 1,6-dioxygenase complexed with substrate and with an inhibitor
Authors: Li De-Feng , Zhang Jia-Yue , Hou Yanjie , Liu Lei , Liu Shuang-Jiang , Liu Wei
Published: October 2012

Dioxygen activation implemented by nonhaem FeIIenzymes containing the 2-His-1-carboxylate facial triad has been extensively studied in recent years. Extradiol dioxygenase is the archetypal member of this superfamily and catalyzes the oxygenolytic ring opening of catechol analogues. Here, the crystallization and preliminary X-ray analysis of 2-aminophenol 1,6-dioxygenase, an enzyme representing a minor subset of extradiol dioxygenases that catalyze the fission of 2-aminophenol rather than catecholic compounds, is reported. Crystals of the holoenzyme with FeIIand of complexes with the substrate 2-aminophenol and the suicide inhibitor 4-nitrocatechol were grown using the cocrystallization method under the same conditions as used for the crystallization of the apoenzyme. The crystals belonged to space groupC2 and diffracted to 2.3–2.7 Å resolution; the crystal that diffracted to the highest resolution had unit-cell parametersa= 270.24,b= 48.39,c= 108.55 Å, β = 109.57°. All X-ray data sets collected from diffraction-quality crystals were suitable for structure determination.
Expression, purification, crystallization and preliminary X-ray diffraction analysis of the dihydroorotase domain of human CAD
Authors: Lallous Nada , Grande-García Araceli , Molina Rafael , Ramón-Maiques Santiago
Published: October 2012

CAD is a 243 kDa eukaryotic multifunctional polypeptide that catalyzes the first three reactions ofde novopyrimidine biosynthesis: glutamine-dependentcarbamyl phosphate synthetase,aspartate transcarbamylase anddihydroorotase (DHO). In prokaryotes, these activities are associated with monofunctional proteins, for which crystal structures are available. However, there is no detailed structural information on the full-length CAD protein or any of its functional domains apart from that it associates to form a homohexamer of ∼1.5 MDa. Here, the expression, purification and crystallization of the DHO domain of human CAD are reported. The DHO domain forms homodimers in solution. Crystallization experiments yielded small crystals that were suitable for X-ray diffraction studies. A diffraction data set was collected to 1.75 Å resolution using synchrotron radiation at the SLS, Villigen, Switzerland. The crystals belonged to the orthorhombic space groupC2221, with unit-cell parametersa= 82.1,b= 159.3,c= 61.5 Å. The Matthews coefficient calculation suggested the presence of one protein molecule per asymmetric unit, with a solvent content of 48%.
Expression, purification, crystallization and preliminary X-ray crystallographic analysis of Zucchini fromDrosophila melanogaster
Authors: Fukuhara Satoshi , Nishimasu Hiroshi , Bonnefond Luc , Matsumoto Naoki , Ishitani Ryuichiro , Nureki Osamu
Published: October 2012

PIWI-interacting RNAs (piRNAs) bind PIWI proteins and silence transposons to maintain the genomic integrity of germ cells. Zucchini (Zuc), a phospholipase D superfamily member, is conserved among animals and is implicated in piRNA biogenesis. However, the underlying mechanism by which Zuc participates in piRNA biogenesis remains elusive.Drosophila melanogasterZuc (DmZuc) was expressed inEscherichia coli, purified and crystallized. X-ray diffraction data were collected to 1.75 Å resolution. The crystal belonged to space groupP21, with unit-cell parametersa= 55.0,b= 71.2,c= 56.3 Å, β = 107.9°.
Production and crystallization of a panel of structure-based mutants of the human myelin peripheral membrane protein P2
Authors: Lehtimäki Mari , Laulumaa Saara , Ruskamo Salla , Kursula Petri
Published: October 2012

The myelin sheath is a multilayered membrane that surrounds and insulates axons in the nervous system. One of the proteins specific to the peripheral nerve myelin is P2, a protein that is able to stack lipid bilayers. With the goal of obtaining detailed information on the structure–function relationship of P2, 14 structure-based mutated variants of human P2 were generated and produced. The mutants were designed to potentially affect the binding of lipid bilayers by P2. All mutated variants were also crystallized and preliminary crystallographic data are presented. The structural data from the mutants will be combined with diverse functional assays in order to elucidate the fine details of P2 function at the molecular level.
Crystallization, optimization and preliminary X-ray characterization of a metal-dependent PI-PLC fromStreptomyces antibioticus
Authors: Jackson Michael R. , Selby Thomas L.
Published: October 2012

A recombinant metal-dependent phosphatidylinositol-specific phospholipase C (PI-PLC) fromStreptomyces antibioticushas been crystallized by the hanging-drop method with and without heavy metals. The native crystals belonged to the orthorhombic space groupP222, with unit-cell parametersa= 41.26,b= 51.86,c = 154.78 Å. The X-ray diffraction results showed significant differences in the crystal quality of samples soaked with heavy atoms. Additionally, drop pinning, which increases the surface area of the drops, was also used to improve crystal growth and quality. The combination of heavy-metal soaks and drop pinning was found to be critical for producing high-quality crystals that diffracted to 1.23 Å resolution.
Cloning, expression, purification, crystallization and preliminary X-ray crystallographic analyses of threonyl-tRNA synthetase editing domain fromAeropyrum pernix
Authors: Ahmad Sadeem , Sravankumar Antony S. K. , Kruparani Shobha P. , Sankaranarayanan Rajan
Published: October 2012

The proofreading function of aminoacyl-tRNA synthetases is crucial in maintaining the fidelity of protein synthesis. Most archaeal threonyl-tRNA synthetases (ThrRSs) possess a unique proofreading domain unrelated to their eukaryotic/bacterial counterpart. The crystal structure of this domain from the archaeonPyrococcus abysiiin complex with its cognate and noncognate substrate analogues had given insights into its catalytic and discriminatory mechanisms. To probe further into the mechanistic and evolutionary aspects of this domain, work has been extended to another archaeonAeropyrum pernix. The organism possesses two proteins corresponding to threonyl-tRNA synthetase,i.e.ThrRS1 and ThrRS2, encoded by two different genes,thrS1andthrS2, respectively. ThrRS1 is responsible for aminoacylation and ThrRS2 for proofreading activity. Here the purification, crystallization and preliminary X-ray crystallographic investigation of the N-terminal proofreading domain of ThrRS2 fromA. pernixis reported. The crystals belong to either theP41212 orP43212 space group and consist of one monomer per asymmetric unit.
A comparison of gene expression of Listeria monocytogenes in vitro and in the soft cheese Crescenza
Authors: Alessandria Valentina , Rantsiou Kalliopi , Dolci Paola , Zeppa Giuseppe , Cocolin Luca
Published: October 2012

This study focuses on the expression analysis of virulence and stress response genes (plcA, hly, iap and sigB) in 11 different strains of Listeria monocytogenes. The first step of this study considered an in vitro analysis in synthetic laboratory medium. According to statistical analysis, significant differences emerged for genes sigB and plcA. Subsequently, a soft cheese at two temperatures (4 and 12 °C) and for different times (24 and 48 h) was evaluated. Significant expression differences emerged for the condition at 12 °C after 48 h. No association could be observed between gene expression profile and origin of the different strains.
A short note on how tied fights affect cortisol levels
Authors: COOPER Edwin L. , NGO Kathy T.
Published: October 2012

Previous work exploring the interrelationships between sex steroids (e.g. androgens, testosterones and 11‐ketotestosterones) and social behavior in teleosts suggest that mirror‐elicited aggression in cichlid fish may not trigger a hormonal response. Using the Mozambique tilapia (Oreochromis mossambicus) to analyze immune responses as a result of social stress, we measured levels of cortisol and melatonin using Enzyme‐Linked Immunosorbent Assay (ELISA) assays. In this work, we demonstrated that cortisol concentrations are significantly lower yet the levels of melatonin remain unchanged in tilapia that are fighting their mirror image. Our results suggested that in tied fights, certain hormone levels remain unchanged (e.g. androgens) due to the lack of melatonin induction.
The Dilmun temple on Failaka, Kuwait
Authors: Højlund Flemming
Published: October 2012

The Kuwaiti‐Danish 2009 excavations on Failaka produced new evidence for the dating of the Dilmun temple in Tell F6. The affinities of this temple to the Barbar temples in Bahrain are emphasised, whereas the hypothesis of a Syrian‐inspired temple tower is questioned.
An Umm an‐Nar culture stone bas‐relief from Al‐Qutainah, Sultanate of Oman
Authors: Böhme Manfred , Al‐Bakri Sultan Saif Nasser
Published: October 2012

The Al‐Qutainah stone with its bas‐relief decoration was found in a secondary position, reused through time. The object is clearly a facing stone, originally part of an Umm an‐Nar period tomb, and was subsequently repeatedly reused for making new engravings. While working out the sequence of images it became clear that the very first image was a pair of human figures. This artwork is associated to grave architecture and therefore dates to the Umm an‐Nar period. Two models of interpretation can be suggested, the image of a human pair or the image of a supplicant and deity. Changes in the original meaning of the image indicate conceptual changes in subsequent periods, most probably alterations in the underlying social structure. The importance of the Qutainah stone is not solely due to rare monumental human figurative expression of the period in Oman; furthermore, the find spot in the Adam region is at a distance from the central location of Umm an‐Nar culture that has so far emerged.
A metabolic signature predicts biological age in mice
Authors: Tomás‐Loba Antonia , Bernardes de Jesus Bruno , Mato Jose M. , Blasco Maria A.
Published: October 2012

Our understanding of the mechanisms by which aging is produced is still very limited. Here, we have determined the sera metabolite profile of 117 wild‐type mice of different genetic backgrounds ranging from 8 to 129 weeks of age. This has allowed us to define a robust metabolomic signature and a derived metabolomic score that reliably/accurately predicts the age of wild‐type mice. In the case of telomerase‐deficient mice, which have a shortened lifespan, their metabolomic score predicts older ages than expected. Conversely, in the case of mice that overexpress telomerase, their metabolic score corresponded to younger ages than expected. Importantly, telomerase reactivation late in life by using a TERT‐based gene therapy recently described by us significantly reverted the metabolic profile of old mice to that of younger mice, further confirming an anti‐aging role for telomerase. Thus, the metabolomic signature associated with natural mouse aging accurately predicts aging produced by telomere shortening, suggesting that natural mouse aging is in part produced by presence of short telomeres. These results indicate that the metabolomic signature is associated with the biological age rather than with the chronological age. This constitutes one of the first aging‐associated metabolomic signatures in a mammalian organism.
Species richness increases the resilience of wetland plant communities in a tropical floodplain
Authors: CARVALHO PRISCILLA , THOMAZ SIDINEI MAGELA , KOBAYASHI JOSILAINE TAECO , BINI LUIS MAURICIO
Published: October 2012

In the last two decades, the relationship between diversity and stability/ecosystem functioning has been widely discussed and has become a central issue in ecology. Here, we assessed the relationship between wetland plant diversity and community resilience after a disturbance. Our study area was located in theUpperParanáRiver floodplain (Brazil). An experiment was carried outin situ(18 1 m × 1 m plots with richness varying from 1 to 18 species). In each plot, we recorded the number of species, total per cent vegetation cover and per cent age cover of each species. The above‐ground biomass of wetland plants was removed, simulating a disturbance by animal trampling or an extreme flood. The recovery of vegetation was monitored over 3 months. According to a linear regression, the recovery of wetland plants was positively correlated with diversity. Comparisons with plots containing monocultures of one of the dominant species (Polygonum stelligerum) suggested that this species did not overyield in mixed cultures. Thus, our experiments indicate that the higher resilience in richer plots after a disturbance is mainly due to the fact that species have different resource use requirements (complementarity effect) and not due to the presence of a single, more productive species. Our experiment carried out in a more real condition (in situ) showed that biodiversity is important to wetland functioning and stability, paralleling the results obtained in laboratory and mesocosms experiments. These results also suggest that the loss of plant diversity in our study area could compromise community recovery following strong disturbances.
Implementation of a clinical practice guideline for antenatal magnesium sulphate for neuroprotection in Australia and New Zealand
Authors: Bain Emily , Bubner Tanya , Ashwood Pat , Crowther Caroline A. , Middleton Philippa ,   The WISH Project Team
Published: October 2012

Health professionals at 25 Australian and New Zealand tertiary maternity hospitals were surveyed about local implementation of a clinical practice guideline for antenatal magnesium sulphate for fetal neuroprotection. Seventy‐six percent of respondents reported that their hospital is currently following a guideline; 36% confirmed that their hospital is auditing uptake. Estimates of uptake ranged from 53 to 90%. Ongoing education and support are needed to ensure that the guidelines are optimally implemented, and uptake and important health outcomes are monitored.
Atopic dermatitis 2.0: from the clinical phenotype to the molecular taxonomy and stratified medicine
Authors: Bieber Th.
Published: October 2012

Atopic dermatitis (AD) is a paradigmatic inflammatory chronic skin disease. As for other chronic skin diseases, (i) the spectrum of the clinical phenotype and severity as well as (ii) the genetic background and (iii) the underlying mechanisms strongly suggest a high degree of pathophysiological heterogeneity yet leading to a similar clinical pattern, that is, the eczematous skin lesion, but showing distinct progression patterns. This review suggests to exploit the recent knowledge about AD for a novel approach proposing a tentative first molecular taxonomy of this disease based on the genotype and endophenotype. The consequences in terms of personalized prevention and management are delineated.
Incidence, clinical characteristics and outcome in Norwegian children with periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis syndrome; a population‐based study
Authors: Førsvoll Jostein , Kristoffersen Einar Klæboe , Øymar Knut
Published: October 2012

Aim

To describe the incidence, epidemiology, clinical presentation and clinical outcome of children with the syndrome of periodic fever, aphthous stomatitis, pharyngitis and cervical adenitis (PFAPA) in a population‐based study.

Methods

In a prospective population‐based study, all children in South Rogaland, Norway, diagnosed with PFAPA during 2004–2010 were evaluated clinically, and parents were interviewed systematically. A follow‐up interview was performed for all patients.

Results

A total of 46 children (32 boys; p = 0.011) were diagnosed with PFAPA. We calculated an incidence of 2.3 per 10 000 children up to 5 years of age. The median age of onset was 11.0 months (quartiles: 5.0, 14.8). Nearly 37 children were followed until resolution. In 17 of these, a tonsillectomy was performed with prompt resolution of PFAPA in all. The median age of spontaneous resolution was 60.2 months (range 24–120) and in children with tonsillectomy 50.9 months (range 15–128).

Conclusion

The incidence of PFAPA was 2.3 per 10 000 children up to 5 years of age. In the majority of cases, onset of symptoms may be during the first year of life.
A descriptive study of older bipolar disorder residents living in New York City’s adult congregate facilities
Authors: Sheeran Thomas , Greenberg Rebecca L , Davan Laura A , Dealy Jennifer A , Young Robert C , Bruce Martha L
Published: October 2012

Sheeran T, Greenberg RL, Davan LA, Dealy JA, Young RC, Bruce ML. A descriptive study of older bipolar disorder residents living in New York City’s adult congregate facilities.  Bipolar Disord 2012: 14: 756–763. © 2012 John Wiley & Sons A/S.

Objectives:  Much of the research on geriatric bipolar disorder is from outpatient populations or epidemiological surveys with small samples. In contrast, in this study a descriptive analysis was conducted of geriatric and younger adult residents with bipolar disorder or mania in non‐clinical adult congregate facilities (ACFs) in the greater New York City region.

Methods:  A total of 2602 ACF residents were evaluated in 19 facilities, across multiple demographic and health domains. Within this sample, 200 residents had chart diagnoses of bipolar disorder or mania. Among these, 50 geriatric residents (age ≥ 60) were compared to 50 younger adult residents (age < 50) on a number of demographic and health measures.

Results:  Based on chart diagnoses, the overall prevalence of bipolar disorder was 7.8%. Compared to other studies of outpatient, epidemiological, and census samples, both older and younger residents with bipolar disorder had higher rates of cognitive impairment, impairment in executive functioning, vision impairment, and proportion of residents who had never been married. The younger group also had higher rates of obesity and the elderly group had a greater proportion of residents without high‐school education. Both age groups had rates of lithium or valproate use comparable to those found in outpatient studies. Comparing the two age groups, the elderly sample had lower overall cognitive and executive functioning, and was using a larger number of medication classes than the younger group. The elderly group also had a larger proportion of residents who were separated/divorced or widowed compared to the younger group, which had higher rates of never‐married residents.

Conclusions:  Overall, both age groups had relatively high rates of bipolar disorder, with significant cognitive impairment, medical burden, obesity, mental health service use, and lower education levels, as compared to outpatient, epidemiological, and census samples. Of note was the significant cognitive impairment across age groups.
Lamotrigine as add‐on treatment to lithium and divalproex: lessons learned from a double‐blind, placebo‐controlled trial in rapid‐cycling bipolar disorder
Authors: Kemp David E , Gao Keming , Fein Elizabeth B , Chan Philip K , Conroy Carla , Obral Sarah , Ganocy Stephen J , Calabrese Joseph R
Published: October 2012

Kemp DE, Gao K, Fein EB, Chan PK, Conroy C, Obral S, Ganocy SJ, Calabrese JR. Lamotrigine as add‐on treatment to lithium and divalproex: lessons learned from a double‐blind, placebo‐controlled trial in rapid‐cycling bipolar disorder.  Bipolar Disord 2012: 14: 780–789. © 2012 John Wiley & Sons A/S.

Objectives:  A substantial portion of the morbidity associated with rapid‐cycling bipolar disorder (RCBD) stems from refractory depression. This study assessed the antidepressant effects of lamotrigine as compared with placebo when used as add‐on therapy for rapid‐cycling bipolar depression non‐responsive to the combination of lithium plus divalproex.

Methods:  During Phase 1 of this trial, hypomanic, manic, mixed, and/or depressed outpatients (n = 133) aged 18–65 years with DSM–IV RCBD type I or II were initially treated with the open combination of lithium and divalproex for up to 16 weeks. During Phase 2, subjects who did not meet the criteria for stabilization (n = 49) (i.e., remained in or cycled into the depressed phase) were randomly assigned to double‐blind, adjunctive lamotrigine (n = 23) or adjunctive placebo (n = 26). The primary endpoint was the mean change in depression symptom severity from the beginning of Phase 2 to the end of Phase 2 (week 12) on the Montgomery‐Åsberg Depression Rating Scale (MADRS) total score. Data were analyzed by analysis of covariance with last observation carried forward and a mixed‐models analysis.

Results:  During Phase 1, a high rate of study discontinuations occurred due to intolerable side effects (13/133; 10%) and study non‐adherence (22/133; 17%). Only 14% (19/133) stabilized on the open combination of lithium and divalproex. Among the 49 (37%) patients randomized to the double‐blind adjunctive treatment phase, mean ± standard error change from baseline on the MADRS total score was −8.5 ± 1.7 points for lamotrigine and −9.1 ± 1.5 points for placebo (p = not significant; mixed‐models analysis). No significant differences were observed in the rates of response, remission, or bimodal response between lamotrigine and placebo.

Conclusions:  The poor tolerability, lack of efficacy, and high rate of early discontinuation with the combination of lithium and divalproex suggests this regimen was ineffective for the majority of patients with RCBD. Among patients who did not stabilize on lithium and divalproex, the addition of lamotrigine was no more effective than placebo in reducing depression severity. The findings suggest an opportunity for several design modifications to enhance signal detection in future trials of RCBD. The main limitation is the small number of subjects randomized to double‐blind treatment.
Ethics in action: Consent‐gaining interactions and implications for research practice
Authors: Speer Susan A. , Stokoe Elizabeth
Published: October 2012

This article deals with the topic of social psychological research methods in practice, by examining how informed consent is gained from research participants. In most research, the consent‐gaining process is hidden from analytic scrutiny and is dealt with before data collection has begun. In contrast, conversation analytic research, which records interactional encounters from beginning to end, enables examination of this methodological ‘black box’. We explored how ‘requests’ to consent in research played out across different institutional settings. We found that participants had to ‘opt‐out’ of a research process that was already underway. Consent‐gaining sequences constrained opting out in two ways: (1) because research activity was already underway, it must be stopped affirmatively by participants; (2) consent‐gaining turns were tilted in favour of continued participation, making opting out a dispreferred response. We also found a mismatch between what ethics guidelines specify about consent‐gaining ‘in theory’ and what actually happens ‘in practice’. Finally, we make suggestions about interventions in and recommendations for existing practice to best achieve informed consent.
Extracellular matrix rigidity controls podosome induction in microvascular endothelial cells
Authors: Juin Amélie , Planus Emmanuelle , Guillemot Fabien , Horakova Petra , Albiges‐Rizo Corinne , Génot Elisabeth , Rosenbaum Jean , Moreau Violaine , Saltel Frédéric
Published: October 2012

Background information

Podosomes are actin‐based structures involved in cell adhesion, migration, invasion and extracellular matrix degradation. They have been described in large vessel endothelial cells, but nothing is known concerning microvascular endothelial cells. Here, we focussed on liver sinusoidal endothelial cells (LSECs), fenestrated microvascular cells that play major roles in liver physiology. Liver fibrosis induces a dedifferentiation of LSECs leading notably to a loss of fenestrae. Because liver fibrosis is associated with increased matrix stiffness, and because substrate stiffness is known to regulate the actin cytoskeleton, we investigated the impact of matrix rigidity on podosome structures in LSECs.

Results

Using primary LSECs, we demonstrated that microvascular endothelial cells are able to form constitutive podosomes. Podosome presence in LSECs was independent of cytokines such as transforming growth factor‐β or vascular endothelial growth factor, but could be modulated by matrix stiffness. As expected, LSECs lost their differentiated phenotype during cell culture, which was paralleled by a loss of podosomes. LSECs however retained the capacity to form active podosomes following detachment/reseeding or actin‐destabilising drug treatments. Finally, constitutive podosomes were also found in primary microvascular endothelial cells from other organs.

Conclusions

Our results show that microvascular endothelial cells are able to form podosomes without specific stimulation. Our data suggest that the major determinant of podosome induction in these cells is substrate rigidity.
Demographic, socioeconomic, and behavioral factors affecting patterns of tooth decay in the permanent dentition: principal components and factor analyses
Authors: Shaffer John R. , Polk Deborah E. , Feingold Eleanor , Wang Xiaojing , Cuenco Karen T. , Weeks Daniel E. , DeSensi Rebecca S. , Weyant Robert J. , Crout Richard , McNeil Daniel W. , Marazita Mary L.
Published: October 2012

Objectives

Dental caries of the permanent dentition is a multifactorial disease resulting from the complex interplay of endogenous and environmental risk factors. The disease is not easily quantitated due to the innumerable possible combinations of carious lesions across individual tooth surfaces of the permanent dentition. Global measures of decay, such as the DMFS index (which was developed for surveillance applications), may not be optimal for studying the epidemiology of dental caries because they ignore the distinct patterns of decay across the dentition. We hypothesize that specific risk factors may manifest their effects on specific tooth surfaces leading to patterns of decay that can be identified and studied. In this study, we utilized two statistical methods of extracting patterns of decay from surface‐level caries data to create novel phenotypes with which to study the risk factors affecting dental caries.

Methods

Intra‐oral dental examinations were performed on 1068 participants aged 18–75 years to assess dental caries. The 128 tooth surfaces of the permanent dentition were scored as carious or not and used as input for principal components analysis (PCA) and factor analysis (FA), two methods of identifying underlying patterns without a priori knowledge of the patterns. Demographic (age, sex, birth year, race/ethnicity, and educational attainment), anthropometric (height, body mass index, waist circumference), endogenous (saliva flow), and environmental (tooth brushing frequency, home water source, and home water fluoride) risk factors were tested for association with the caries patterns identified by PCA and FA, as well as DMFS, for comparison. The ten strongest patterns (i.e. those that explain the most variation in the data set) extracted by PCA and FA were considered.

Results

The three strongest patterns identified by PCA reflected (i) global extent of decay (i.e. comparable to DMFS index), (ii) pit and fissure surface caries and (iii) smooth surface caries, respectively. The two strongest patterns identified by FA corresponded to (i) pit and fissure surface caries and (ii) maxillary incisor caries. Age and birth year were significantly associated with several patterns of decay, including global decay/DMFS index. Sex, race, educational attainment, and tooth brushing were each associated with specific patterns of decay, but not with global decay/DMFS index.

Conclusions

Taken together, these results support the notion that caries experience is separable into patterns attributable to distinct risk factors. This study demonstrates the utility of such novel caries patterns as new outcomes for exploring the complex, multifactorial nature of dental caries.
Immunotherapeutic effects of recombinant adenovirus encoding granulocyte–macrophage colony-stimulating factor in experimental pulmonary tuberculosis
Authors: Francisco-Cruz A , Mata-Espinosa D , Estrada-Parra S , Xing Z , Hernández-Pando R
Published: October 2012

BALB/c mice with pulmonary tuberculosis (TB) develop a T helper cell type 1 that temporarily controls bacterial growth. Bacterial proliferation increases, accompanied by decreasing expression of interferon (IFN)-γ, tumour necrosis factor (TNF)-α and inducible nitric oxide synthase (iNOS). Activation of dendritic cells (DCs) is delayed. Intratracheal administration of only one dose of recombinant adenoviruses encoding granulocyte–macrophage colony-stimulating factor (AdGM-CSF) 1 day before Mycobacterium tuberculosis (Mtb) infection produced a significant decrease of pulmonary bacterial loads, higher activated DCs and increased expression of TNF-α, IFN-γ and iNOS. When AdGM-CSF was given in female mice B6D2F1 (C57BL/6J X DBA/2J) infected with a low Mtb dose to induce chronic infection similar to latent infection and corticosterone was used to induce reactivation, a very low bacilli burden in lungs was detected, and the same effect was observed in healthy mice co-housed with mice infected with mild and highly virulent bacteria in a model of transmissibility. Thus, GM-CSF is a significant cytokine in the immune protection against Mtb and gene therapy with AdGM-CSF increased protective immunity when administered in a single dose 1 day before Mtb infection in a model of progressive disease, and when used to prevent reactivation of latent infection or transmission.
Effect of interleukin-6 receptor blockade on the balance between regulatory T cells and T helper type 17 cells in rheumatoid arthritis patients
Authors: Pesce B , Soto L , Sabugo F , Wurmann P , Cuchacovich M , López M N , Sotelo P H , Molina M C , Aguillón J C , Catalán D
Published: October 2012

A new paradigm has emerged relating the pathogenesis of rheumatoid arthritis (RA), focused on the balance between T helper type 17 cells and regulatory T cells (Tregs). In humans, both subpopulations depend on transforming growth factor (TGF)-β for their induction, but in the presence of inflammatory cytokines, such as interleukin (IL)-6, the generation of Th17 is favoured. Tocilizumab is a therapeutic antibody targeting the IL-6 receptor (IL-6R), which has demonstrated encouraging results in RA. The aim of this study was to evaluate the effect of tocilizumab on Th1 cells, Th17 cells, IL-17 and interferon (IFN)-γ double secretors Th17/Th1 cells, and Tregs in RA patients. Eight RA patients received tocilizumab monthly for 24 weeks and blood samples were obtained every 8 weeks to study T cell populations by flow cytometry. The frequency of Th17 cells, Th1 cells and Th17/Th1 cells was evaluated in peripheral blood mononuclear cells (PBMCs) activated in vitro with a polyclonal stimulus. Tregs were identified by their expression of forkhead box protein 3 (FoxP3) and CD25 by direct staining of PBMCs. Although no changes were detected in the frequency of Th1 or Th17 cells, the percentages of peripheral Tregs increased after therapy. In addition, the infrequent Th17/Th1 subpopulation showed a significant increment in tocilizumab-treated patients. In conclusion, tocilizumab was able to skew the balance between Th17 cells and Tregs towards a more protective status, which may contribute to the clinical improvement observed in RA patients.
Bone tissue microarchitectural characteristics at dental implant sites part 2: correlation with bone classification and primary stability
Authors: Ribeiro‐Rotta Rejane Faria , de Oliveira Rubelisa Cândido Gomes , Dias Danilo Rocha , Lindh Christina , Leles Cláudio Rodrigues
Published: October 2012

Objective

To investigate in vivo the correlation between the bone microarchitecture of implant bone sites, bone tissue classification subjectively assessed in radiographs and primary implant stability.

Material and methods

Periapical and panoramic radiographs were obtained from 32 partially edentulous patients. Three surgeons classified bone quality at implant sites using two different methods: assessments in periapical and panoramic radiographs (PP) and according to the classification proposed by Lekholm and Zarb (L&Z). During the implant insertion, bone biopsies were taken, and three‐dimensional parameters were measured by microcomputed tomography (microCT). Insertion torque value (ITV) and initial implant stability quotient (ISQ) were recorded at the moment of the implantation. ISQ was also recorded at the uncovering stage of a traditional implant protocol.

Results

Bone types 2 and 3 were the most prevalent classifications according to PP (54.3%) and L&Z (58.7%). The mean and standard deviation values of primary stability variables were 38.7(16.7) for ITV, 75.3 (7.7) for initial ISQ and 79.3 (6.8) for uncovering ISQ. Several microCT original and factor variables were found to correlate with bone tissue classifications and primary stability variables. L&Z correlated with architecture (r = 0.31; P < 0.05), density (r = −0.43; P < 0.01) and bulk (r = −0.35; P < 0.05), whereas ITV correlated with architecture (r = −0.40; P < 0.01) and density (r = 0.51; P < 0.01). Multiple linear regression analysis revealed that density and bulk explained 32% of the variability of L&Z bone classification, while density and architecture explained 42% of the variability of ITV.

Conclusions

This study demonstrates the first step in validating L&Z classification. MicroCT provides objective and detailed quantitative data on bone microarchitecture. Intraosseous implant stability is mainly determined by the density of the bone. ITVs could be a good indicator of primary implant stability, whereas ISQs measurements have some limitations and should not be used alone.
In vitro assessment of artifacts induced by titanium dental implants in cone beam computed tomography
Authors: Benic Goran I. , Sancho‐Puchades Manuel , Jung Ronald E. , Deyhle Hans , Hämmerle Christoph H.F.
Published: October 2012

Objective

The objective of this study was to evaluate the geometric pattern and the intensity of artifacts around titanium implants in cone beam computed tomography (CBCT) using an in vitro model.

Material and methods

Ten test models, each containing one 4.1‐mm‐diameter titanium implant, were cast from a human mandible using silicone impression material and dental stone. Each model contained an implant in one of the following single‐tooth gaps: 37, 36, 34, 33, 31, 41, 43, 44, 46, and 47. For control purposes, three models without implants were produced. Each model was scanned five times using a CBCT scanner. Gray values (GV) were recorded at eight circumferential positions around the implants at 0.5 mm, 1 mm, and 2 mm from the implant surface (GVTest). GV were measured in the corresponding volumes of interest (VOI) in the models without implants (GVControl). Differences of gray values (ΔGV) between GVTest and GVControl were calculated as percentages. To detect differences between GVTest and GVControl, the 95% confidence interval (CI) was computed for the values of ΔGV. Repeated measures ANOVA was used for the comparison of ΔGV at 0.5 mm, 1 mm, and 2 mm from the implant surface.

Results

Artifacts reflected by altered GV were always present in the proximity of titanium implants, regardless of the implant position. When comparing GVTest and GVControl, increased GV were found at the buccal and lingual aspects of the implant sites, whereas regions with reduced GV were located along the long axis of the mandibular body of the test models. A significant decrease in artifact intensity was found with increasing distance from the buccal implant surface (ΔGV0.5 mm: 45 ± 10% [SD], ΔGV1 mm: 28 ± 14% [SD], ΔGV2 mm: 14 ± 7% [SD]) (P < 0.001).

Conclusion

Artifacts around titanium implants in CBCT images were distributed according to a geometrical pattern.
Transient swelling of the Schneiderian membrane after transversal sinus augmentation: a pilot study
Authors: Quirynen Marc , Lefever David , Hellings Peter , Jacobs Reinhilde
Published: October 2012

Objective

The predictability and simplicity of the Summers' technique for sinus floor elevation encouraged many clinicians to consider this new approach. The impact of such intervention on the sinus mucosa has, however, not been explored extensively. This pilot study followed the response of the Schneiderian membrane, longitudinally, via cone‐beam CT images.

Material and methods

Ten consecutive patients (five women, mean age 58.7 years) referred for implant therapy in the posterior part of the maxilla, in combination with a trans‐alveolar sinus floor elevation (13 sinuses involved), were enrolled. CBCT images were taken prior to and at 1 week and 1 month after surgery. The changes in thickness of the Schneiderian membrane were scored at nine standardized points per sinus using reformatted cross‐sectional images.

Results

One week after sinus floor elevation, the Schneiderian membrane showed a significant swelling of 9.2 mm in the middle (SD 0.3; P < 0.005), 7.2 mm medially (SD 2.5; P < 0.005) and 3.9 mm laterally (SD 0,3; P < 0.05), respectively, resulting in an overall mean of 6.7 mm (SD 2.6), which fully disappeared 3 weeks later. This swelling involved the entire floor of the sinus and not only the augmented area.

Conclusions

Within the limitations of this pilot study, one can conclude that the Schneiderian membrane responds with a significant “transient” swelling (5–10× its size) during the first weeks of healing.
Generation of albino Xenopus tropicalis using zinc‐finger nucleases
Authors: Nakajima Keisuke , Nakajima Taeko , Takase Minoru , Yaoita Yoshio
Published: October 2012

To generate albino lines of Xenopus tropicalis, we injected fertilized eggs with mRNAs encoding zinc‐finger nucleases (ZFNs) targeting the tyrosinase coding region. Surprisingly, vitiligo was observed on the skin of F0 frogs that had been injected with ZFN mRNAs, indicating that both tyrosinase genes in the genome were disrupted in all melanocytes within the vitiligo patches. Mutation analysis using genomic DNA from the skin revealed that two mosaic F0 frogs underwent spatially complex tyrosinase gene mutations. The data implies that the ZFN‐induced tyrosinase gene ablations occurred randomly over space and time throughout the entire body, possibly until the young tadpole stage, and that melanocyte precursors lacking functional tyrosinase proliferated and formed vitiligo patches. Several albino X. tropicalis, which are compound heterozygotes for biallelic tyrosinase mutations, were obtained by mating the mosaic F0 frogs. To our knowledge, this is the first report of the albino vertebrates generated by the targeted gene knockout.
Direct and indirect effects of a thrips‐transmitted Tospovirus on the preference and fitness of its vector, Frankliniella fusca
Authors: Shrestha Anita , Srinivasan Rajagopalbabu , Riley David G. , Culbreath Albert K.
Published: October 2012

Phytoviruses including tospoviruses are known to affect the behavior and fitness of their vectors both positively and negatively. In this study, we investigated the effects of Tomato spotted wilt virus (TSWV) (family Bunyaviridae, genus Tospovirus) infection on the fitness and feeding ability of tobacco thrips, Frankliniella fusca (Hinds) (Thysanoptera: Thripidae) using peanut, Arachis hypogaea L. (Fabaceae), as a host. Potentially viruliferous F. fusca laid more eggs than non‐viruliferous F. fusca. In contrast, fewer potentially viruliferous F. fusca developed into adults and required a longer developmental time than non‐viruliferous F. fusca, indicating a direct negative effect of the virus on thrips fitness. In addition, no‐choice feeding tests indicated that non‐viruliferous F. fusca fed more rapidly than potentially viruliferous F. fusca. Typically, phytovirus infections are known to enhance the availability of vital nutrients such as free amino acids in infected host plants and to affect other important physiological processes negatively. Free amino acids are known to play a vital role in egg production and development. Further investigations in this study revealed that leaflets of infected plants had ca. 15 times more free amino acids than non‐infected leaflets. TSWV‐infected leaflets were used to rear potentially viruliferous thrips. Higher amino acid levels in TSWV‐infected leaflets than in non‐infected leaflets could have contributed to increased oviposition by potentially viruliferous F. fusca compared to non‐viruliferous F. fusca. Taken together, these results suggest that increased concentrations of free amino acids in TSWV‐infected plants might serve as an incentive for thrips feeding on otherwise unsuitable hosts, thereby facilitating TSWV acquisition and transmission.
Imitating others' actions: transcranial magnetic stimulation of the parietal opercula reveals the processes underlying automatic imitation
Authors: Mengotti Paola , Ticini Luca F. , Waszak Florian , Schütz‐Bosbach Simone , Rumiati Raffaella I.
Published: October 2012

The human tendency to imitate gestures performed by conspecifics is automatic in nature. However, whether this automatic imitation can be considered as a true imitative phenomenon or only as a special instance of spatial compatibility is still being debated. New evidence suggests that automatic imitation, otherwise known as ‘imitative compatibility’, shall be considered as a phenomenon that operates independently from spatial compatibility. So far there are only a few investigations directly aimed at identifying the neural structures dedicated to this process. In the present study, we applied double‐pulse transcranial magnetic stimulation (TMS) over the parietal opercula to further investigate the role of these regions in coding imitative compatibility. We found that a temporary disruption of parietal opercula caused the reduction of the imitative compatibility relative to the sham condition. In particular, the TMS interference with the parietal opercula's activity modulated the imitative compatibility but not the spatial compatibility, suggesting that these two processes are likely to be independent.