bluuebunny/crossref_metadata_2025 huggingface dataset:
Quality of Health Care in the United States: Implications for Pediatric Inflammatory Bowel Disease
Authors: Boyle Brendan M , Palmer Lena , Kappelman Michael D
Published: August 2009

The Institute of Medicine's publications To Error is Human and Crossing the Quality Chasm publicized the widespread deficits in US health care quality. Emerging studies continue to reveal deficits in the quality of adult and pediatric care, including subspecialty care. In recent years, key stakeholders in the health care system including providers, purchasers, and the public have been applying various quality improvement methods to address these concerns. Lessons learned from these efforts in other pediatric conditions, including asthma, cystic fibrosis, neonatal intensive care, and liver transplantation may be applicable to the care of children with inflammatory bowel disease (IBD).

This review is intended to be a primer on the quality of care movement in the United States, with a focus on pediatric IBD. In this article, we review the history, rationale, and methods of quality measurement and improvement, and we discuss the unique challenges in adapting these general strategies to pediatric IBD care.
Releasing cohesin from chromosome arms in early mitosis: opposing actions of Wapl–Pds5 and Sgo1
Authors: Shintomi Keishi , Hirano Tatsuya
Published: August 2009

The cohesin complex establishes sister chromatid cohesion during S phase. In metazoan cells, most if not all cohesin dissociates from chromatin during mitotic prophase, leading to the formation of metaphase chromosomes with two cytologically discernible chromatids. This process, known as sister chromatid resolution, is believed to be a prerequisite for synchronous separation of sister chromatids in subsequent anaphase. To dissect this process at a mechanistic level, we set up an in vitro system. Sister chromatid resolution is severely impaired upon depletion of Wapl from Xenopus egg extracts. Exogenously added human Wapl can rescue these defects and, remarkably, it can do so in a very short time window of early mitosis. A similar set of observations is made for Pds5, a factor implicated previously in the stabilization of interphase cohesion. Characteristic amino acid motifs (the FGF motifs) in Wapl coordinate its physical and functional interactions with Pds5 and cohesin subunits. We propose that Wapl and Pds5 directly modulate conformational changes of cohesin to make it competent for dissociation from chromatin during prophase. Evidence is also presented that Sgo1 plays a hitherto underappreciated role in stabilizing cohesin along chromosome arms, which is antagonized by the mitotic kinases polo-like kinsase (Plk1) and aurora B.
The human protein coevolution network
Authors: Tillier Elisabeth R.M. , Charlebois Robert L.
Published: August 2009

Coevolution maintains interactions between phenotypic traits through the process of reciprocal natural selection. Detecting molecular coevolution can expose functional interactions between molecules in the cell, generating insights into biological processes, pathways, and the networks of interactions important for cellular function. Prediction of interaction partners from different protein families exploits the property that interacting proteins can follow similar patterns and relative rates of evolution. Current methods for detecting coevolution based on the similarity of phylogenetic trees or evolutionary distance matrices have, however, been limited by requiring coevolution over the entire evolutionary history considered and are inaccurate in the presence of paralogous copies. We present a novel method for determining coevolving protein partners by finding the largest common submatrix in a given pair of distance matrices, with the size of the largest common submatrix measuring the strength of coevolution. This approach permits us to consider matrices of different size and scale, to find lineage-specific coevolution, and to predict multiple interaction partners. We used MatrixMatchMaker to predict protein–protein interactions in the human genome. We show that proteins that are known to interact physically are more strongly coevolving than proteins that simply belong to the same biochemical pathway. The human coevolution network is highly connected, suggesting many more protein–protein interactions than are currently known from high-throughput and other experimental evidence. These most strongly coevolving proteins suggest interactions that have been maintained over long periods of evolutionary time, and that are thus likely to be of fundamental importance to cellular function.
Clinicopathological study of Fox–Fordyce disease
Authors: KAO Pei‐Han , HSU Chao‐Kai , LEE Julia Yu‐Yun
Published: August 2009

Fox–Fordyce disease (FFD) is a rare skin disease manifesting as multiple pruritic follicular papules involving the skin‐bearing apocrine glands. Reports of FFD in Asian people are scant. In this retrospective study, we describe the clinicopathological findings of five cases of FFD affecting Taiwanese subjects. Clinically, all patients presented with numerous uniform, 2–3‐mm, skin‐colored to light brown, dome‐shaped papules with smooth surface, which were distributed in the apocrine gland‐containing areas. Pruritus varied from mild to severe. The histopathology is characterized by focal spongiosis in the upper infundibulum with perifollicular fibrosis and lymphohistiocytic infiltrate. FFD needs to be differentiated from lichen amyloidosis, Darier’s disease, syringoma, lichen simplex chronicus and spongiotic dermatitis clinically or pathologically. The findings of focal spongiosis in upper infundibulum associated with a perifollicular lymphohistiocytic infiltrate can facilitate the diagnosis of FFD.
Intraspecies transmission of L‐type‐like bovine spongiform encephalopathy detected in Japan
Authors: Fukuda Shigeo , Iwamaru Yoshifumi , Imamura Morikazu , Masujin Kentarou , Shimizu Yoshihisa , Matsuura Yuichi , Shu Yujing , Kurachi Megumi , Kasai Kazuo , Murayama Yuichi , Onoe Sadao , Hagiwara Ken’ichi , Sata Tetsutaro , Mohri Shirou , Yokoyama Takashi , Okada Hiroyuki
Published: August 2009

It has been assumed that the agent causing BSE in cattle is a uniform strain (classical BSE); however, different neuropathological and molecular phenotypes of BSE (atypical BSE) have been recently reported. We demonstrated the successful transmission of L‐type‐like atypical BSE detected in Japan (BSE/JP24 isolate) to cattle. Based on the incubation period, neuropathological hallmarks, and molecular properties of the abnormal host prion protein, the characteristics of BSE/JP24 prion were apparently distinguishable from the classical BSE prion and closely resemble those of bovine amyloidotic spongiform encephalopathy prion detected in Italy.
TP53 deletion is not an adverse feature in multiple myeloma treated with total therapy 3
Authors: Shaughnessy John D. , Zhou Yiming , Haessler Jeff , Van Rhee Frits , Anaissie Elias , Nair Bijay , Waheed Sarah , Alsayed Yazan , Epstein Joshua , Crowley John , Barlogie Bart
Published: August 2009

Contrary to Total Therapy (TT) 2 for multiple myeloma patients, FGFR3− translocation bore no adverse effects on outcome in TT3 with added bortezomib. Del TP53, another poor‐risk feature in TT2 and present in 10% of 441 patients treated, was examined for its prognostic consequences in TT3. Not affecting rate or duration of complete response, TP53 haplo‐insufficiency also did not compromise, in the 83% with genomically defined low‐risk myeloma, survival or event‐free survival. FGFR3+ and FGFR3− molecular subgroups fared worse in the presence of del TP53 when applying TT2 but not TT3. Thus, the prognostic implications of del TP53 were protocol‐, genome‐defined risk‐ and molecular subgroup‐dependent.
The impact of caring for a child with cerebral palsy: quality of life for mothers and fathers
Authors: Davis E. , Shelly A. , Waters E. , Boyd R. , Cook K. , Davern M.
Published: August 2009

Background  Although it is expected that caring for a child with cerebral palsy (CP) can impact on the quality of life (QOL) of caregivers, the QOL of carers' has yet to be adequately examined. The aims of this study are to: (1) explore the QOL of mothers and fathers of children with CP aged 3–18 years; and (2) examine whether the impact of caring for a child with CP changes from childhood to adolescence.

Method  A qualitative study was conducted utilizing a grounded theory framework. Twenty‐four mothers and 13 fathers of children and adolescents with CP aged 3–7 years (n = 15), 8–12 years (n = 10) and 13–18 years (n = 12) and with varying levels of impairment (GMFCS Level I = 1, II = 4, II = 3, IV = 5, V = 12) participated in semi‐structured interviews about their QOL. The transcripts were analysed to identify issues affecting parental QOL.

Results  There were no differences in parental QOL among subgroups (i.e. mothers and fathers, age groups, GMFCS levels). Parental QOL ranged across a wide spectrum. Caring for a child with CP affects a parent's physical well‐being, social well‐being, freedom and independence, family well‐being and financial stability. Parents indicated that they often feel unsupported by the services they access.

Conclusions  Caring for a child with CP can both positively and negatively impact on a parent's life. There is value for both parents and children if parental concerns and determinants of QOL are considered in overall programme planning and service delivery for children and their families.
A comparison of Rasch item‐fit and Cronbach's alpha item reduction analysis for the development of a Quality of Life scale for children and adolescents
Authors: Erhart M. , Hagquist C. , Auquier P. , Rajmil L. , Power M. , Ravens‐Sieberer U. ,   the European KIDSCREEN Group
Published: August 2009

Objective  This study compares item reduction analysis based on classical test theory (maximizing Cronbach's alpha – approach A), with analysis based on the Rasch Partial Credit Model item‐fit (approach B), as applied to children and adolescents' health‐related quality of life (HRQoL) items. The reliability and structural, cross‐cultural and known‐group validity of the measures were examined.

Methods  Within the European KIDSCREEN project, 3019 children and adolescents (8–18 years) from seven European countries answered 19 HRQoL items of the Physical Well‐being dimension of a preliminary KIDSCREEN instrument. The Cronbach's alpha and corrected item total correlation (approach A) were compared with infit mean squares and the Q‐index item‐fit derived according to a partial credit model (approach B). Cross‐cultural differential item functioning (DIF ordinal logistic regression approach), structural validity (confirmatory factor analysis and residual correlation) and relative validity (RV) for socio‐demographic and health‐related factors were calculated for approaches (A) and (B).

Results  Approach (A) led to the retention of 13 items, compared with 11 items with approach (B). The item overlap was 69% for (A) and 78% for (B). The correlation coefficient of the summated ratings was 0.93. The Cronbach's alpha was similar for both versions [0.86 (A); 0.85 (B)]. Both approaches selected some items that are not strictly unidimensional and items displaying DIF. RV ratios favoured (A) with regard to socio‐demographic aspects. Approach (B) was superior in RV with regard to health‐related aspects.

Conclusion  Both types of item reduction analysis should be accompanied by additional analyses. Neither of the two approaches was universally superior with regard to cultural, structural and known‐group validity. However, the results support the usability of the Rasch method for developing new HRQoL measures for children and adolescents.
Influences of biological risk at birth and temperament on development at toddler and preschool ages
Authors: Hwang A‐W. , Soong W‐T. , Liao H‐F.
Published: August 2009

Purposes  This study aimed at investigating: (1) the effects of temperament and biological risk at birth on various developmental domains at toddler and preschool ages; (2) the interaction effects of the biological risk and temperament on development.

Participants  One hundred and ten full‐term and 98 preterm children without significant physical or developmental disabilities and consisting of various biological risks were examined at toddler age (18–36 months) and preschool age (51–67 months).

Methods  The Neonatal Medical Index was used to classify the biological risk level. Parental reports on the Chinese Toddler Temperament Scale at toddler age were collected and the temperament (easy, intermediate and difficult) of each child was assigned according to local norm. The Comprehensive Developmental Inventory for Infants and Toddlers was used for assessing child development at toddle and preschool ages. Other family variables were also collected as possible confounders for child development. Two‐way ancova was conducted to analyse the predictability of biological risk and temperament, by controlling the potential family variables on child development.

Results  At toddler age, higher biological risk had significant adverse effects on both the Perceptual‐motor developmental quotients (DQs) (F1,201= 19.4, P < 0.001) and Social‐adaptive DQs (F2,200= 22.5, P < 0.001), while easy temperament had positive effects only on the Social‐adaptive DQs (F2,200= 7.7, P < 0.01). At preschool age, none of the two factors had effects on DQs of the Comprehensive Developmental Inventory for Infants and Toddlers. There were no significant interactions between biological risk and temperament on DQs at both ages.

Conclusion  The biological risk and temperament affected child development at toddler age but not at preschool age. No interaction of biological risk and temperament effects on the child development at toddler age existed. The effects of biological risk and temperament on child development were temporary for the children with relatively low biological risk.
Input of organochlorine compounds by snow to European high mountain lakes
Authors: GRIMALT JOAN O. , FERNÁNDEZ PILAR , QUIROZ ROBERTO
Published: August 2009

1. Persistent organic pollutants (POPs) can be trapped by and accumulate in cold regions. To understand POP accumulation in temperate high mountain lakes, we collected samples of snow from the catchments of several high mountain lakes in Europe.

2. Organochlorine compounds (OCs) are regularly found in snow collected in European high mountain sites. Polychlorobiphenyls (PCBs) were found in all samples examined. Hexachlorocyclohexanes (HCHs) are also common whereas DDTs and hexachlorobenzene were found less frequently.

3. Comparison of the concentrations of these pollutants in snow with OC levels in the waters of these lakes or atmospheric deposition during cold periods shows that the snowpack constitutes a significant seasonal PCB reservoir in all catchments and that snow trapping is a major mechanism for the incorporation of HCHs in the lakes situated in the coldest sites, e.g. Gossenköllesee in the Alps.

4. Correlation of the concentrations of the PCB congeners in snow versus mean annual winter temperature shows higher accumulation at lower temperatures. The less volatile PCBs exhibit higher temperature dependences than the more volatile congeners. This trend differs from altitudinal dependences observed in other sites such as Canada and may be related to the specific range of winter temperature in each area of study.
The effects of morphology and substrate diameter on climbing and locomotor performance in male spiders
Authors: Prenter John , Pérez‐Staples Diana , Taylor Phillip W.
Published: August 2009

1. Spiders are the most sexually size dimorphic terrestrial animals and the evolution of this dimorphism is controversial. Patterns of sexual size dimorphism (SSD) in spiders have been related to individual performance and size. In 2002 Moya‐Laraño, Halaj & Wise proposed the ‘gravity hypothesis’ to explain patterns of sexual size dimorphism in spiders whereby species building webs high in the vegetation are predicted to show greater SSD than those that build lower down. They advocated an advantage in climbing speed in smaller males searching for females in high places. The gravity hypothesis predicts a negative relationship between male size and climbing speed. In 2007 Brandt & Andrade questioned this interpretation and proposed that the pattern of SSD in spiders is better explained by an advantage for larger males of low‐dwelling species to run faster along the ground.

2. We induced male spiders to run a standard distance up vertical poles of different diameters to examine the predicted relationship between size and climbing speed. We tested two species of extremely size‐dimorphic orb‐web spiders, Argiope keyserlingi and Nephila plumipes, that differ in the height at which females tend to build webs, and one species of jumping spider, Jacksonoides queenslandica, with low levels of size dimorphism. We also examined morphological determinants of horizontal motility by inducing males to run along a raceway.

3. Substrate diameter was consistently found to influence climbing performance. In N. plumipes, climbing speed was slowest on the widest diameter substrate. In A. keyserlingi, size‐adjusted leg length and substrate diameter interacted to determine climbing speed, while in J. queenslandica, there was an interaction between body size and substrate diameter on climbing speed. In the effect of substrate diameter, we have identified a potential bias in previous tests of the gravity hypothesis.

4. Our results do not support the prediction of the gravity hypothesis. There was no evidence of a negative relationship between body size and climbing speed in the two orb‐web species with high levels of SSD. Our results are also not consistent with a recent modification of the gravity hypothesis that suggests a curvilinear relationship between climbing speed and size.

5. Body size was positively associated with maximum running speed only in the cursorial hunter J. queenslandica. For this spider, results are more consistent with Brandt & Andrade’s explanation for variation in SSD in spiders, that larger males are selected for superior running ability in low‐dwelling species, rather than selection for smaller size for climbing to females in high‐dwelling species.
Are increased concentrations of maternal corticosterone adaptive to offspring? A test using a placentotrophic lizard
Authors: Cadby Chloé D. , Jones Susan M. , Wapstra Erik
Published: August 2009

1. Maternal hormones during embryogenesis act as a bridge between the maternal and the offspring environment and consequently may allow maternal pre‐programming of offspring phenotype to the environment, assuming that maternal environment is a reliable predictor of offspring environment.

2. We use an orthogonal experimental design in which we increase plasma corticosterone concentration (vs. control) in placentotrophic gravid female spotted skinks (Niveoscincus ocellatus) and their offspring. We hypothesize that high concentrations of maternal corticosterone allow offspring phenotype to be pre‐programmed to better cope with high concentrations of corticosterone after birth and, consequently, offspring would not suffer from its deleterious effects. We use growth as a measure of performance as corticosterone typically affects growth in reptiles.

3. High concentrations of maternal corticosterone had significant effects on offspring body condition at birth which could have resulted from corticosterone passing through the complex placenta and/or from the indirect effects of corticosterone on maternal body condition.

4. There was no indication of maternal pre‐programming to high concentrations of corticosterone postpartum: growth rate of offspring placed in the same treatment (corticosterone vs. control) than their mother was not different to growth rate of offspring placed in a different treatment than their mother.

5. Maternal pre‐programming to corticosterone may not have been selected for in this species either because maternal chronic stress is not a reliable predictor of offspring chronic stress or because maternal pre‐programming does not outweigh the costs of having reduced sensitivity to the hormone, resulting in slower, less appropriate reactions in stressful conditions. This study further demonstrates the highly versatile and context‐dependent nature of maternal effects and the trade‐offs between costs and benefits of maternal pre‐programming.
Foundations of communities of practice: enablers and barriers to participation
Authors: Guldberg K. , Mackness J.
Published: August 2009

This research draws upon community of practice theory to explore the factors that enabled or hindered participation in an online ‘Foundations of Communities of Practice’ workshop – a course that is designed to align with Wenger's communities of practice perspective. The research used a mixed methods approach, drawing upon log‐on and posting data, questionnaires and semi‐structured interviews to explore participant experiences. The findings show that five dimensions either enabled or constrained participation. These were emotion, technology, connectivity, understanding norms and learning tensions. As enablers these dimensions led to successful participation within an online community of practice, but as constraints, they led to peripheral participation. The findings highlight implications for tutors of such courses. These include the need to (1) assess the technical expertise of participants, particularly when a number of different technological tools are used; (2) find ways to identify and evaluate emotional responses so learners can be supported in managing these; (3) ensure that participants understand the norms of a community; and (4) develop clear induction materials and processes.
A review of high‐pressure freezing preparation techniques for correlative light and electron microscopy of the same cells and tissues
Authors: MCDONALD K.L.
Published: August 2009

In this paper, we review some published studies using correlative light and electron microscopy methods. We further refined our criteria to include only those studies using live cells for light microscope and where high‐pressure freezing was the method of specimen preparation for electron microscopy. High‐pressure freezing is especially important for some difficult‐to‐fix samples, and for optimal preservation of ultrastructure in samples larger than a few micrometres. How the light microscope observations are done is completely sample dependent, but the choice of high‐pressure freezer depends on the speed required to capture (freeze) the biological event of interest. For events requiring high time resolution (in the 4–5 s range) the Leica EM PACT2 with rapid transfer system works well. For correlative work on structures of interest that are either non‐motile or moving slowly (minutes rather than seconds), any make of high‐pressure freezer will work. We also report on some efforts to improve the capabilities of the Leica EM PACT2 rapid transfer system.
Molecular cryo‐electron tomography of vitreous tissue sections: current challenges
Authors: NORLÉN L. , ÖKTEM O. , SKOGLUND U.
Published: August 2009

Electron tomography of vitreous tissue sections (tissue TOVIS) allows the study of the three‐dimensional structure of molecular complexes in a near‐native cellular context. Its usage is, however, limited by an unfortunate combination of noisy and incomplete data, by a technically demanding sample preparation procedure, and by a disposition for specimen degradation during data collection. Here we outline some major challenges as experienced from the application of TOVIS to human skin. We further consider a number of practical measures as well as theoretical approaches for its future development.
Correlative fluorescence and electron microscopy in tissues: immunocytochemistry
Authors: ROBINSON J.M. , TAKIZAWA T.
Published: August 2009

Correlative microscopy is a collection of procedures that rely upon two or more imaging modalities to examine the same specimen. The imaging modalities employed should each provide unique information and the combined correlative data should be more information rich than that obtained by any of the imaging methods alone. Currently the most common form of correlative microscopy combines fluorescence and electron microscopy. While much of the correlative microscopy in the literature is derived from studies of model cell culture systems we have focused, primarily, on correlative microscopy in tissue samples. The use of tissue, particularly human tissue, may add constraints not encountered in cell culture systems. Ultrathin cryosections, typically used for immunoelectron microscopy, have served as the substrate for correlative fluorescence and electron microscopic immunolocalization in our studies. In this work, we have employed the bifunctional reporter FluoroNanogold. This labeling reagent contains both a fluorochrome and a gold‐cluster compound and can be imaged by sequential fluorescence and electron microscopy. This approach permits the examination of exactly the same sub‐cellular structures in both fluorescence and electron microscopy with a high level of spatial resolution.
Correlative microscopy: a potent tool for the study of rare or unique cellular and tissue events
Authors: MIRONOV A.A. , BEZNOUSSENKO G.V.
Published: August 2009

Biological studies have relied on two complementary microscope technologies – light (fluorescence) microscopy and electron microscopy. Light microscopy is used to study phenomena at a global scale to look for unique or rare events, and it also provides an opportunity for live imaging, whereas the forte of electron microscopy is the high resolution. Traditionally light and electron microscopy observations are carried out in different populations of cells/tissues and a ‘correlative’ inference is drawn. The advent of true correlative light‐electron microscopy has allowed high‐resolution imaging by electron microscopy of the same structure observed by light microscopy, and in advanced cases by video microscopy. Thus a rare event captured by low‐resolution imaging of a population or transient events captured by live imaging can now also be studied at high resolution by electron microscopy. Here, the potential and difficulties of this approach, along with the most impressive breakthroughs obtained by these methods, are discussed.
Correlative fluorescence and transmission electron microscopy: an elegant tool to study the actin cytoskeleton of whole‐mount (breast) cancer cells
Authors: JAHN K.A. , BARTON D.A. , SU Y. , RICHES J. , KABLE E.P.W. , SOON L.L. , BRAET F.
Published: August 2009

Elucidating the structure and dynamics of lamellipodia and filopodia in response to different stimuli is a topic of continuing interest in cancer cells as these structures may be attractive targets for therapeutic purposes. Interestingly, a close functional relationship between these actin‐rich protrusions and specialized membrane domains has been recently demonstrated. The aim of this study was therefore to investigate the fine organization of these actin‐rich structures and examine how they structurally may relate to detergent‐resistant membrane (DRM) domains in the MTLn3 EGF/serum starvation model. For this reason, we designed a straightforward and alternative method to study cytoskeleton arrays and their associated structures by means of correlative fluorescence (/laser)‐ and electron microscopy (CFEM).

 CFEM on whole mounted breast cancer cells revealed that a lamellipodium is composed of an intricate filamentous actin web organized in various patterns after different treatments. Both actin dots and DRM's were resolved, and were closely interconnected with the surrounding cytoskeleton. Long actin filaments were repeatedly observed extending beyond the leading edge and their density and length varied after different treatments. Furthermore, CFEM also allowed us to demonstrate the close structural association of DRMs with the cytoskeleton in general and the filamentous/dot‐like structural complexes in particular, suggesting that they are all functionally linked and consequently may regulate the cell's fingertip dynamics. Finally, electron tomographic modelling on the same CFEM samples confirmed that these extensions are clearly embedded within the cytoskeletal matrix of the lamellipodium.
Towards correlative imaging of plant cortical microtubule arrays: combining ultrastructure with real‐time microtubule dynamics
Authors: BARTON D.A. , GARDINER J.C. , OVERALL R.L.
Published: August 2009

There are a variety of microscope technologies available to image plant cortical microtubule arrays. These can be applied specifically to investigate direct questions relating to array function, ultrastructure or dynamics. Immunocytochemistry combined with confocal laser scanning microscopy provides low resolution “snapshots” of cortical microtubule arrays at the time of fixation whereas live cell imaging of fluorescent fusion proteins highlights the dynamic characteristics of the arrays. High‐resolution scanning electron microscopy provides surface detail about the individual microtubules that form cortical microtubule arrays and can also resolve cellulose microfibrils that form the innermost layer of the cell wall. Transmission electron microscopy of the arrays in cross section can be used to examine links between microtubules and the plasma membrane and, combined with electron tomography, has the potential to provide a complete picture of how individual microtubules are spatially organized within the cortical cytoplasm. Combining these high‐resolution imaging techniques with the expression of fluorescent cytoskeletal fusion proteins in live cells using correlative microscopy procedures will usher in an radical change in our understanding of the molecular dynamics that underpin the organization and function of the cytoskeleton.
Rapid combined light and electron microscopy on large frozen biological samples
Authors: VOGELS I.M.C. , HOEBEN K.A. , VAN NOORDEN C.J.F.
Published: August 2009

The use of large unfixed frozen tissue samples (10 × 10 × 5 mm3) for combined light microscopy (LM) and electron microscopy (EM) is described. First, cryostat sections are applied for various LM histochemical approaches including in situ hybridization, immunohistochemistry and metabolic mapping (enzyme histochemistry). When EM inspection is needed, the tissue blocks that were used for cryostat sectioning and are stored at −80 °C, are then fixed at 4 °C with glutaraldehyde/paraformaldehyde and prepared for EM according to standard procedures. Ultrastructurally, most morphological aspects of normal and pathological tissue are retained whereas cryostat sectioning at −25 °C does not have serious damaging effects on the ultrastructure. This approach allows simple and rapid combined LM and EM of relatively large tissue specimens with acceptable ultrastructure. Its use is demonstrated with the elucidation of transdifferentiated mouse stromal elements in human pancreatic adenocarcinoma explants grown subcutaneously in nude mice. Combined LM and EM analysis revealed that these elements resemble cartilage showing enchondral mineralization and aberrant muscle fibres with characteristics of skeletal muscle cells.
A case of obstructive sleep apnoea with anterior cervical osteophytes
Authors: ANDO E. , OGAWA T. , SHIGETA Y. , HIRAI S. , IKAWA T. , ISHIKAWA C. , NEJIMA J.
Published: August 2009

Summary Osteophytes of the cervical spine are usually seen in elderly adults. When prominent, they have been blamed for dysphagia, cough, dysphonia and dyspnoea. This paper reports on an obstructive sleep apnoea (OSA) patient with cervical spinal osteophytes, one cause of airway obstruction. A 75‐year‐old male complained of pronounced snoring. The diagnosis was mild OSA, apnoea hypopnoea index was 9·4. Patient reported no restrictions in neck movements, experiences of neck pain or neck trauma. Previously, patient underwent a tonsillectomy due to discomfort in the pharyngeal region. A lateral cephalometric image was taken to observe airway before oral appliance therapy. The image revealed the presence of large osteophytes or sclerotic enthesopathy, lying on anterior surfaces from the fourth to seventh cervical vertebrae. A computed tomography (CT) image revealed the relationship of airway position to the spine. In the reconstructed three‐dimensional (3D) image, the airway appeared displaced to the right of the craniomandiblar bone, with the hyoid bone similarly displaced in a manner to that of the airway. The spine also appeared displaced to the left side ofcraniomandiblar bone. Additionally, the 3D image revealed calcification of the stylohyoideum ligament and ligamentum nuchae. This present case highlights the necessity of CT examination for OSA patients. There were several ligament calcifications in the head and neck region. Cervical spine osteophytes, as a component of Forestier’s or cervical spine disease, have been associated with dysphagia and dysphonia. It was reported that bilateral vocal cord paralysis was caused by osteophytes compressing the post‐cricoid area of larynx.
Influence of material and diameter of pre‐fabricated posts on maxillary central incisors restored with crown
Authors: GONZÁLEZ‐LLUCH C. , RODRÍGUEZ‐CERVANTES P. J. , SANCHO‐BRU J. L. , PÉREZ‐GONZÁLEZ A. , BARJAU‐ESCRIBANO A. , VERGARA‐MONEDERO M. , FORNER‐NAVARRO L.
Published: August 2009

Summary  Numerous research works have studied the effect of post‐design parameters on the mechanical behaviour of restored teeth without reaching any clear conclusions. Previous works by the authors ascertained the effect of material and post‐dimensions for non‐crowned restored teeth. The aim of this work was to study the effect of post‐material and diameter for crowned teeth. First, an experimental fracture strength test was performed on eighteen extracted human maxillary central incisors. Teeth were decoronated, treated endodontically and restored (nine with glass fibre posts and nine with stainless steel posts). Several post‐diameters were used. The final crown restoration was carried out using a reinforced glass‐ceramic material. Failure loads were recorded and results were compared using the one‐way anova. Secondly, the finite element technique was used to model the restored teeth and to compare the estimated stress distributions. The addition of the crown did not affect the strength of the restoration to any significant extent and post‐diameter did not influence the biomechanical performance of either of the post‐systems. The crown acts as a protector, thus eliminating the influence of the post‐diameter that was found previously when using stainless steel posts, but it does not completely rule out the possibility of a root fracture. Significantly, lower failure loads were found experimentally for teeth restored with stainless steel posts. The stress distributions predicted by the model corroborated these findings and allowed the authors to propose the use of glass fibre posts as a more robust restorative technique.
Parallel habitat‐driven differences in the phylogeographical structure of two independent lineages of Mediterranean saline water beetles
Authors: ABELLÁN P. , MILLÁN A. , RIBERA I.
Published: August 2009

It has been hypothesized that species living in small lentic water bodies, because of the short‐term geological persistence of their habitat, should show higher dispersal ability, with increased gene flow among populations and a less pronounced phylogeographical structure. Conversely, lotic species, living in more geologically stable habitats, should show reduced dispersal and an increased phylogeographical structure at the same geographical scales. In this work we tested the influence of habitat type in two groups of aquatic Coleoptera (Nebrioporus ceresyiandOchthebius notabilisgroups, families Dytiscidae and Hydraenidae respectively), each of them with closely related species typical of lotic and lentic saline Western Mediterranean water bodies. We used mitochondrialcox1sequence data of 453 specimens of 77 populations through the range of nine species to compare a lotic vs. a lentic lineage in each of the two groups. Despite the differences in biology (predators vs. detritivorous) and evolutionary history, in both lotic lineages there was a higher proportion of nucleotide diversity among than within groups of populations, and a faster rate of accumulation of haplotype diversity (as measured by rarefaction curves) than in the lentic lineages. Similarly, lotic lineages had a higher absolute phylogenetic diversity, more remarkable considering their smaller absolute geographical ranges. By comparing closely related species, we were able to show the effect of contrasting habitat preferences in two different groups, in agreement with predictions derived from habitat stability.
Hfq negatively regulates type III secretion in EHEC and several other pathogens
Authors: Shakhnovich Elizabeth A. , Davis Brigid M. , Waldor Matthew K.
Published: August 2009

Hfq is a conserved RNA‐binding protein that regulates diverse cellular processes through post‐transcriptional control of gene expression, often by functioning as a chaperone for regulatory sRNAs. Here, we explored the role of Hfq in enterohaemorrhagic Escherichia coli (EHEC), a group of non‐invasive intestinal pathogens. EHEC virulence is dependent on a Type III secretion system encoded in the LEE pathogenicity island. The abundance of transcripts for all 41 LEE genes and more than half of confirmed non‐LEE‐encoded T3 effectors were elevated in an EHEC hfq deletion mutant. Thus, Hfq promotes co‐ordinated expression of the LEE‐encoded T3S apparatus and both LEE‐ and non‐LEE‐encoded effectors. Increased transcript levels led to the formation of functional secretion complexes capable of secreting high quantities of effectors into the supernatant. The increase in LEE‐derived transcripts and proteins was dependent on Ler, the LEE‐encoded transcriptional activator, and the ler transcript appears to be a direct target of Hfq‐mediated negative regulation. Finally, we found that Hfq contributes to the negative regulation of T3SSs in several other pathogens, suggesting that Hfq, potentially along with species‐specific sRNAs, underlies a common means to prevent unfettered expression of T3SSs.
The TviA auxiliary protein renders the Salmonella enterica serotype Typhi RcsB regulon responsive to changes in osmolarity
Authors: Winter Sebastian E. , Winter Maria G. , Thiennimitr Parameth , Gerriets Valerie A. , Nuccio Sean‐Paul , Rüssmann Holger , Bäumler Andreas J.
Published: August 2009

In response to osmolarity, Salmonella enterica serotype Typhi (S. Typhi) regulates genes required for Vi capsular antigen expression oppositely to those required for motility and invasion. Previous studies suggest that osmoregulation of motility, invasion and capsule expression is mediated through the RcsC/RcsD/RcsB phosphorelay system. Here we performed gene expression profiling and functional studies to determine the role of TviA, an auxiliary protein of the RcsB response regulator, in controlling virulence gene expression in S. Typhi. TviA repressed expression of genes encoding flagella and the invasion‐associated type III secretion system (T3SS‐1) through repression of the flagellar regulators flhDC and fliZ, resulting in reduced invasion, reduced motility and reduced expression of FliC. Both RcsB and TviA repressed expression of flhDC, but only TviA altered flhDC expression in response to osmolarity. Introduction of tviA into S. enterica serotype Typhimurium rendered flhDC transcription sensitive to changes in osmolarity. These data suggest that the auxiliary TviA protein integrates a new regulatory input into the RcsB regulon of S. Typhi, thereby altering expression of genes encoding flagella, the Vi antigen and T3SS‐1 in response to osmolarity.
Impact of nadir lower oesophageal sphincter pressure on bolus clearance assessed by combined manometry and multi‐channel intra‐luminal impedance measurement
Authors: Nguyen N. Q. , Ching K. , Tippett M. , Smout A. J. P. M. , Holloway R. H.
Published: August 2009

Abstract  This study aimed to assess the relationship between nadir lower oesophageal sphincter pressure (LOSP) and wave amplitude (WA) in oesophageal bolus clearance. Concurrent oesophageal manometry and impedance were performed in 146 subjects [41 healthy, 24 non‐obstructive dysphagia (NOD) and 81 gastro‐oesophageal reflux (GOR)]. Patients with achalasia and diffuse oesophageal spasm were excluded. Swallow responses were categorized by nadir LOSP. For each category of nadir LOSP, WA at the distal 2 recording sites were grouped into bins of 10 mmHg and the proportion of waves in each bin associated with a normal bolus presence time (BPT) was determined. Nadir LOSP, distal BPT, total bolus transit time and the proportion of impaired oesophageal clearance in patients with NOD were greater than those of healthy subjects and patients with GOR. Overall, responses with impaired oesophageal clearance had significantly lower WA (54 ± 1 vs 81 ± 1 mmHg; P < 0.0001) and higher nadir LOSP (2.7 ± 0.4 vs 1.0 ± 0.1 mmHg, P < 0.001). For each level of nadir LOSP, there was a direct relationship between distal WA and successful bolus clearance of both liquid and viscous boluses from the distal oesophagus. As nadir LOSP increased, the relationship between WA and bolus clearance shifted to the right and higher amplitudes were required to achieve the same effectiveness of clearance. Hypotensive responses with nadir LOSP ≥3 mmHg were less likely to clear than those with nadir LOSP <3 mmHg, for both liquid (7/29 vs 162/276; P < 0.001) or viscous boluses (11/46 vs 176/279; P < 0.0001). Nadir LOSP is an important determinant of bolus clearance from the distal oesophagus, particularly in patients with NOD.
Serum Soluble Interleukin‐2 Receptor, Beta2‐Microglobulin, Lactate Dehydrogenase and Erythrocyte Sedimentation Rate in Children with Hodgkin’s Lymphoma
Authors: Bien E. , Balcerska A.
Published: August 2009

The study was to determine clinical utility of serum soluble interleukin (IL)‐2 receptor (sIL‐2Rα), β2‐microglobulin (β2‐M), lactate dehydrogenase (LDH) and erythrocyte sedimentation rate (ESR) as markers of diagnosis, prognosis and monitoring of response to therapy in childhood Hodgkin’s lymphoma (HL). The markers were measured prospectively before treatment and in complete remission (CR) during and after therapy in 30 children with HL (F/M:19/11; median age: 11.3 years) and once in 50 healthy children (F/M: 24/26; median age: 8.7 years). Median pretreatment levels of all analysed markers were significantly higher than in healthy controls. Increased pretreatment sIL‐2Rα, LDH and ESR correlated with bulky disease; sIL‐2Rα, β2‐M and ESR with presence of B symptoms and sIL‐2Rα and LDH with advanced HL stages. There was a correlation between sIL‐2Rα and LDH and between β2‐M and ESR. The levels and rates of elevated markers reflected well the response to chemotherapy, decreasing significantly when patients achieved CR and further on with therapy continuation. Since all patients survived thus the markers’ value to predict the outcome was not established. Serum sIL‐2Rα, β2‐M, LDH and ESR may act as markers for diagnostics and used in monitoring of therapy effectiveness in childhood HL. The markers were also increased in subgroups of patients with unfavourable clinical features; however, small sample size of the study did not allow to draw conclusion on their prognostic roles. We were also not able to establish the influence of markers on event free survival and overall survival because all children survived independent of initial clinical characteristics and pretreatment levels of sIL‐2Rα, β2‐M, LDH and ESR.
A survey of physicians' reasons to transfuse plasma and platelets in the critically ill: a prospective single‐centre cohort study
Authors: Vlaar A. P. J. , In Der Maur A. L. , Binnekade J. M. , Schultz M. J. , Juffermans N. P.
Published: August 2009

summary.

Data on the rationality of transfusion practice of fresh frozen plasma (FFP) and platelets in the critically ill are sparse and may contribute to efforts to reduce transfusion rates. To provide insight into determinants of the decision of intensive care unit (ICU)‐physicians to transfuse, a survey study was performed. The reasons of ICU‐physicians to transfuse FFP and platelets were determined during a 10‐week period. Transfusion triggers were assessed, as well as correction of prolonged coagulation test results. Of 310 admissions, 44 patients (14%) received a transfusion of FFP and 35 patients (11%) received a platelet transfusion. In 67% patients, FFPs were transfused in bleeding patients and in 33% in non‐bleeding patients. FFP was transfused at a prothrombin time (PT) of 19 s (17–22). After FFP transfusion, PT levels of 15–18, 18–20 and 20–26 s decreased with a median of 0.7, 1.9 and 3.5 s, respectively. On average, 3.2 FFP units were ordered, of which 28% was not transfused. The major reason to transfuse platelets was bleeding. Platelets were transfused at a platelet count of 95 (36–116) × 109 L−1 in bleeding and 13 (10–18) × 109 L−1 in non‐bleeding patients. On average, 1.4 platelet units were ordered, of which 20% was not transfused. The agreement between physicians reporting a major bleeding and a definition of bleeding was poor (κ < 0.10 for FFP and 0.20 for platelets). In conclusion, one‐third of FFP transfusions was given to non‐bleeding patients. FFP transfusion failed to normalize prolonged coagulation test results in the majority of the patients. Transfusion of platelets was restrictive in non‐bleeding patients and liberal in bleeding patients. Education on indications of FFP transfusion and improved identification of bleeding may reduce transfusion rates.
Nitric oxide and pain: ‘Something old, something new’
Authors: MICLESCU A. , GORDH T.
Published: August 2009

Challenges have emerged following the revival of nitric oxide (NO) from ‘something old’, a simple gas derived from nitrogen and oxygen with a role in the early stages of evolution, into ‘something new’, an endogenously formed biological mediator regulating a wide variety of physiological functions. Although pain is a common sensation, it encompasses multiple neurobiologic components, of which NO is only one. In pain research, the study of NO is complicated by convoluted problems related mostly to the effects of NO, which are pro‐ or anti‐nociceptive depending on the circumstances. This dual function reflects the multi‐faceted roles of the NO molecule described in physiology. This review covers current information about NO and its implications in pain mechanisms. In addition, it follows the pain pathways, demonstrating the role of NO in peripheral nociceptive transmission as well in central sensitization. This knowledge may provide the scientific basis for developing new drugs that are indicated for different types of pain, drugs that may be related to the chemical links of NO. A comprehensive approach to understanding the effects of NO will help clinicians identify novel agents that combine the pharmacological profile of native drugs with a controllable manner of NO release. Inhibitors of NO synthesis may have analgesic effects and would be of interest for treating inflammatory and neuropathic pain. Unfortunately, only a few of these compounds have reached the stage of clinical pain trials.