bluuebunny/crossref_metadata_2025 huggingface dataset:
Demographic and clinical features of systemic sclerosis patients with anti‐RNA polymerase III antibodies
Authors: Motegi Sei‐ichiro , Toki Sayaka , Yamada Kazuya , Uchiyama Akihiko , Ishikawa Osamu
Published: December 2014

Anti‐RNA polymerase III antibody (RNAP) is primarily detected in diffuse cutaneous type systemic sclerosis (dcSSc) patients and strongly associated with renal crisis. Additionally, there has been increasing evidence that cancer in SSc patients is associated with RNAP. The aim of this study was to examine the demographic and clinical features of SSc patients with RNAP. Among 246 SSc patients, 5.7% were positive for RNAP, 20.7% were positive for anti‐topoisomerase I antibody (Topo I) alone and 39.4% were positive for anticentromere antibody (ACA) alone. The modified Rodnan total skin score (mRTSS) in SSc patients with RNAP (19.1 ± 2.6) was significantly higher than those in SSc patients with Topo I (11.5 ± 1.1) and patients with ACA (4.4 ± 0.4). Furthermore, among SSc patients with RNAP, the levels of RNAP were positively correlated with mRTSS. Renal crisis is also significantly more prevalent in SSc patients with RNAP than patients without RNAP. Male sex, dcSSc subtype, digital vasculopathy, including digital ulcers and acro‐osteolysis, interstitial lung disease and rheumatoid arthritis complications were prevalent in SSc patients with RNAP and patients with Topo‐I. Primary biliary cirrhosis and Sjögren's syndrome were more in SSc patients with RNAP and patients with ACA compared with patients with Topo 1. No significant difference in the frequency of complications, including Raynaud's phenomenon, pulmonary artery hypertension and malignancy was observed between the three groups. Thus, measurement of RNAP in SSc patients is useful for the diagnosis and risk stratification of severe manifestation, such as renal crisis and severe skin sclerosis.
Epidemiology of atopic dermatitis in Poland according to the Epidemiology of Allergic Disorders in Poland (ECAP) study
Authors: Sybilski Adam J. , Raciborski Filip , Lipiec Agnieszka , Tomaszewska Aneta , Lusawa Adam , Samel‐Kowalik Piotr , Walkiewicz Artur , Krzych‐Fałta Edyta , Samoliński Bolesław
Published: December 2014

Epidemiological studies (International Study of Asthma and Allergies in Childhood [ISAAC], The European Community Respiratory Health Survey [ECRHS]) revealed considerable geographical differences in prevalence of atopic dermatitis (AD). The aim of our study is to present the epidemiology and the risk factors of AD in a Polish population. A total of 18 617 subjects were selected. The sample included respondents in eight cities and one rural area each over 150 000 citizens. The study had two parts: (i) questionnaire survey conducted among all respondents (response rate, 64.4%); and (ii) allergological examination performed among 25.7% with skin prick test (SPT) with 15 aeroallergens. AD was identified in 3.91% of subjects, more often in females, living in the cities, with a mother and/or father with atopy and with a higher education and higher economic status. Comorbidities of AD were atopic rhinitis (AR) in 26.17% and AR and asthma in 14.6%. AD was diagnosed by allergologists in 311 subjects (6.5%). Positive SPT occurred in 66.9% persons with AD (house dust mite, 33.1%; grasses/crop plants, 30.6%). Of the patients, 9.5% with perennial versus 9.3% seasonal and 9.6% with polyvalent versus 9.0% monovalent sensitization had AD. AD prevalence in Poland is below the mean rate for Europe, but risk factor profile is similar to other countries. AD is more frequent in female, well‐educated individuals, of high socioeconomic status, with atopic parents and who live in a city. Seasonal and monovalent atopy play a more essential role in subjects with AD compared with AR and asthma.
The offset‐midpoint travel‐time pyramid for P‐wave in 2D transversely isotropic media with a tilted symmetry axis
Authors: Hao Qi , Stovas Alexey
Published: December 2014

For pre‐stack phase‐shift migration in homogeneous isotropic media, the offset‐midpoint travel time is represented by the double‐square‐root equation. The travel time as a function of offset and midpoint resembles the shape of Cheops’ pyramid. This is also valid for transversely isotropic media with a vertical symmetry axis. In this study, we extend the offset‐midpoint travel‐time pyramid to the case of 2D transversely isotropic media with a tilted symmetry axis. The P‐wave analytical travel‐time pyramid is derived under the assumption of weak anelliptical property of the tilted transverse isotropy media. The travel‐time equation for the dip‐constrained transversely isotropic model is obtained from the depth‐domain travel‐time pyramid. The potential applications of the derived offset‐midpoint travel‐time equation include pre‐stack Kirchhoff migration, anisotropic parameter estimation, and travel‐time calculation in transversely isotropic media with a tilted symmetry axis.
Regional Governance Networks: Filling In or Hollowing Out?
Authors: Jacobsen Dag Ingvar
Published: December 2014

The debate on what effect networks have on the existing institutions of representative democracy is highly polarized. On one hand, networks are regarded as means to ‘fill in’ where traditional institutions fail, and on the other, networks are viewed as arrangements draining or ‘hollowing out’ traditional institutions of power and legitimacy. In this article, filling in and hollowing out are treated as two distinct dimensions, opening up for the possibility that networks may both fill in and hollow out member organizations. The argument is that networks, to be able to fill in, must be ‘strong’ (autonomy, resources), but the stronger they become, the higher the probability that they also hollow out their individual members. This thesis is empirically illuminated by studying political governance networks inNorway (i.e., voluntary cooperation between municipalities creating a regional council). Main findings conclude that these networks only to a very moderate degree fill in and hollow out, indicating that the networks play a largely symbolic role. However, there are variations between networks, and the data indicate that the degree of hollowing out depends on both the strength of the network (size and asymmetry) and its individual members (i.e., their population size).
Election Inversions under Proportional Representation
Authors: Miller Nicholas R.
Published: December 2014

It has been shown by Peter Kurrild‐Klitgaard, using several empirical examples under the Danish electoral system, that proportional representation (PR) can produce ‘election inversions’ such that a coalition of parties collectively supported by a majority of voters fails to win a majority of parliamentary seats. However, Kurrild‐Klitgaard's examples result from imperfections in the Danish PR system introduced to serve goals other than proportionality. In this article, Kurrild‐Klitgaard's analysis is carried a step further by showing that election inversions can occur even under the purest type of PR – namely, one with (i) a single national constituency, (ii) no explicit seat threshold, and (iii) a highly proportional electoral formula. Inversions result from the unavoidable ‘whole number problem’. Recent election data from Israel and the Netherlands is examined and examples of inversions under their relatively pure PR systems are found. Inversions are also found after recalculating seat allocations without a threshold, and on the basis of the most proportional electoral formulas and when the analysis is restricted to seat‐winning parties. Kurrild‐Klitgaard's Danish data is then re‐examined in the same fashion, as is the most recent apportionment of seats in the United States House of Representatives, and more examples of inversions are found.
Spillover Effects of the Affordable Care Act? Exploring the Impact on Young Adult Dental Insurance Coverage
Authors: Shane Dan M. , Ayyagari Padmaja
Published: December 2014

Objectives

To assess whether the Affordable Care Act's (ACA) dependent coverage health insurance mandate had a spillover impact on young adult dental insurance coverage and whether any observed effects varied by household income.

Data

Medical Expenditure Panel Surveys from 2006 through 2011.

Study Design

We employed a difference‐in‐difference regression approach comparing changes in insurance rates for young adults ages 19–25 years to changes in insurance rates for adults ages 27–30 years. Separate regressions were estimated by categories of household income as a percentage of the Federal Poverty Level (FPL) to understand whether the mandate had heterogeneous spillover effects.

Results

Private dental insurance increased by 6.7 percentage points among young adults compared to a control group of 27–30‐year olds. Increases were concentrated at middle‐income levels (125–400 percent FPL).

Conclusions

The dependent coverage mandate provision of the Affordable Care Act has not only increased health insurance rates among young adults but also dental insurance coverage rates.
Does a Physician's Attitude toward a Patient with Mental Illness Affect Clinical Management of Diabetes? Results from a Mixed‐Method Study
Authors: Welch Lisa C. , Litman Heather J. , Borba Christina P.C. , Vincenzi Brenda , Henderson David C.
Published: December 2014

Objective

To determine whether physician's attitudes toward patients with comorbid mental illness affect management of a chronic disease.

Data Source

A total of 256 primary care physicians interviewed in 2010.

Study Design

This randomized factorial experiment entailed physicians observing video vignettes of patient‐actors with poorly controlled diabetes. Patients were balanced across age, gender, race, and comorbidity (schizophrenia with bizarre or normal affect, depression, eczema).

Data Collection

Physicians completed structured and semistructured interviews plus chart notes about clinical management and attitudes.

Principal Findings

Physicians reported more negative attitudes for patients with schizophrenia with bizarre affect (SBA). There were few differences in clinical decisions measured quantitatively or in charting, but qualitative data revealed less trust of patients with SBA as reporters, with more reliance on sources other than engaging the patient in care. Physicians often alerted colleagues about SBA, thereby shaping expectations before interactions occurred.

Conclusions

Results are consistent with common stereotypes about people with serious mental illness. Vignettes did not include intentional indication of unreliable reporting or danger. Reducing health care disparities requires attention to subtle aspects of managing patients—particularly those with atypical affect—as seemingly slight differences could engender disparate patient experiences over time.
Oscillatory Rheology and Creep Behavior of Barley β‐d‐glucan Concentrate Dough: Effect of Particle Size, Temperature, and Water Content
Authors: Ahmed Jasim , Thomas Linu , Al‐Attar Hasan
Published: December 2014

Small amplitude oscillatory rheology and creep behavior of β‐glucan concentrate (BGC) dough were studied as function of particle size (74, 105, 149, 297, and 595 μm), BGC particle‐to‐water ratio (1:4, 1:5, and 1:6), and temperature (25, 40, 55, 70, and 85 °C). The color intensity and protein content increased with decreasing particle size by creating more surface areas. The water holding capacity (WHC) and sediment volume fraction increased with increasing particle size from 74 to 595 μm, which directly influences the mechanical rigidity and viscoelasticity of the dough. The dough exhibited predominating solid‐like behavior (elastic modulus, G′ > viscous modulus, G″). A discrete retardation spectrum is employed to the creep data to obtain retardation time and compliance parameters, which varied significantly with particle size and the process temperature. Creep tests exhibited more pronounced effect on dough behavior compared to oscillatory measurement. The protein denaturation temperature was insignificantly increased with particle fractions from 107 to 110 °C. All those information could be helpful to identify the particle size range and WHC of BGC that could be useful to produce a β‐d‐glucan enriched designed food.
Use of lectins to in situ visualize glycoconjugates of extracellular polymeric substances in acidophilic archaeal biofilms
Authors: Zhang R. Y. , Neu T. R. , Bellenberg S. , Kuhlicke U. , Sand W. , Vera M.
Published: December 2014

Biofilm formation and the production of extracellular polymeric substances (EPS) by meso‐ and thermoacidophilic metal‐oxidizing archaea on relevant substrates have been studied to a limited extent. In order to investigate glycoconjugates, a major part of the EPS, during biofilm formation/bioleaching by archaea on pyrite, a screening with 75 commercially available lectins by fluorescence lectin‐binding analysis (FLBA) has been performed. Three representative archaeal species, Ferroplasma acidiphilum DSM 28986, Sulfolobus metallicus DSM 6482T and a novel isolate Acidianus sp. DSM 29099 were used. In addition, Acidianus sp. DSM 29099 biofilms on elemental sulfur were studied. The results of FLBA indicate (i) 22 lectins bound to archaeal biofilms on pyrite and 21 lectins were binding to Acidianus sp. DSM 29099 biofilms on elemental sulfur; (ii) major binding patterns, e.g. tightly bound EPS and loosely bound EPS, were detected on both substrates; (iii) the three archaeal species produced various EPS glycoconjugates on pyrite surfaces. Additionally, the substratum induced different EPS glycoconjugates and biofilm structures of cells of Acidianus sp. DSM 29099. Our data provide new insights into interactions between acidophilic archaea on relevant surfaces and also indicate that FLBA is a valuable tool for in situ investigations on archaeal biofilms.
Pathogenic and commensal Escherichia coli from irrigation water show potential in transmission of extended spectrum and AmpC β‐lactamases determinants to isolates from lettuce
Authors: Njage Patrick M. K. , Buys Elna M.
Published: December 2014

There are few studies on the presence of extended‐spectrum β‐lactamases and AmpC β‐lactamases (ESBL/AmpC) in bacteria that contaminate vegetables. The role of the production environment in ESBL/AmpC gene transmission is poorly understood. The occurrence of ESBL/AmpC in Escherichia coli (n = 46) from lettuce and irrigation water and the role of irrigation water in the transmission of resistant E. coli were studied. The presence of ESBL/AmpC, genetic similarity and phylogeny were typed using genotypic and phenotypic techniques. The frequency of β‐lactamase gene transfer was studied in vitro. ESBLs/AmpC were detected in 35 isolates (76%). Fourteen isolates (30%) produced both ESBLs/AmpC. Prevalence was highest in E. coli from lettuce (90%). Twenty‐two isolates (48%) were multi‐resistant with between two and five ESBL/AmpC genes. The major ESBL determinant was the CTX‐M type (34 isolates). DHA (33% of isolates) were the dominant AmpC β lactamases. There was a high conjugation efficiency among the isolates, ranging from 3.5 × 10−2 to 1 × 10−2 ± 1.4 × 10−1 transconjugants per recipient. Water isolates showed a significantly higher conjugation frequency than those from lettuce. A high degree of genetic relatedness between E. coli from irrigation water and lettuce indicated possible common ancestry and pathway of transmission.
A chromosomally encoded T7 RNA polymerase‐dependent gene expression system for Corynebacterium glutamicum: construction and comparative evaluation at the single‐cell level
Authors: Kortmann Maike , Kuhl Vanessa , Klaffl Simon , Bott Michael
Published: December 2014

Corynebacterium glutamicum has become a favourite model organism in white biotechnology. Nevertheless, only few systems for the regulatable (over)expression of homologous and heterologous genes are currently available, all of which are based on the endogenous RNA polymerase. In this study, we developed an isopropyl‐β‐d‐1‐thiogalactopyranosid (IPTG)‐inducible T7 expression system in the prophage‐free strain C. glutamicum MB001. For this purpose, part of the DE3 region of Escherichia coli BL21(DE3) including the T7 RNA polymerase gene 1 under control of the lacUV5 promoter was integrated into the chromosome, resulting in strain MB001(DE3). Furthermore, the expression vector pMKEx2 was constructed allowing cloning of target genes under the control of the T7lac promoter. The properties of the system were evaluated using eyfp as heterologous target gene. Without induction, the system was tightly repressed, resulting in a very low specific eYFP fluorescence (= fluorescence per cell density). After maximal induction with IPTG, the specific fluorescence increased 450‐fold compared with the uninduced state and was about 3.5 times higher than in control strains expressing eyfp under control of the IPTG‐induced tac promoter with the endogenous RNA polymerase. Flow cytometry revealed that T7‐based eyfp expression resulted in a highly uniform population, with 99% of all cells showing high fluorescence. Besides eyfp, the functionality of the corynebacterial T7 expression system was also successfully demonstrated by overexpression of the C. glutamicum pyk gene for pyruvate kinase, which led to an increase of the specific activity from 2.6 to 135 U mg−1. It thus presents an efficient new tool for protein overproduction, metabolic engineering and synthetic biology approaches with C. glutamicum.
Production of 2‐ketoisocaproate with Corynebacterium glutamicum strains devoid of plasmids and heterologous genes
Authors: Vogt Michael , Haas Sabine , Polen Tino , van Ooyen Jan , Bott Michael
Published: December 2014

2‐Ketoisocaproate (KIC), the last intermediate in l‐leucine biosynthesis, has various medical and industrial applications. After deletion of the ilvE gene for transaminase B in l‐leucine production strains of Corynebacterium glutamicum, KIC became the major product, however, the strains were auxotrophic for l‐isoleucine. To avoid auxotrophy, reduction of IlvE activity by exchanging the ATG start codon of ilvE by GTG was tested instead of an ilvE deletion. The resulting strains were indeed able to grow in glucose minimal medium without amino acid supplementation, but at the cost of lowered growth rates and KIC production parameters. The best production performance was obtained with strain MV‐KICF1, which carried besides the ilvE start codon exchange three copies of a gene for a feedback‐resistant 2‐isopropylmalate synthase, one copy of a gene for a feedback‐resistant acetohydroxyacid synthase and deletions of ltbR and iolR encoding transcriptional regulators. In the presence of 1 mM l‐isoleucine, MV‐KICF1 accumulated 47 mM KIC (6.1 g l−1) with a yield of 0.20 mol/mol glucose and a volumetric productivity of 1.41 mmol KIC l−1 h−1. Since MV‐KICF1 is plasmid free and lacks heterologous genes, it is an interesting strain for industrial application and as platform for the production of KIC‐derived compounds, such as 3‐methyl‐1‐butanol.
Endoluminal thoracic aortic repair in the octogenarian and nonagenarian: The New Zealand experience
Authors: Buckenham Tim , Pearch Ben , Wright Isabel
Published: December 2014

Introduction

The aim of this study was to review the New Zealand‐wide experience of thoracic endovascular aortic repair to determine effect of age on outcome.

Methods

This was an observational, retrospective analytic study comparing two age groups. The New Zealand Thoracic Aortic Stent (NZTAS) registry was reviewed for patient demographics, indications for repair, risk factors, technical success, complications, length of hospital stay and in‐hospital mortality.

Results

The 264 patients were divided into two groups: <80 (Group I, n = 245) and >80 years on the day of the procedure (Group II, n = 19). Group II comprised 11 males and 8 females with a mean age (range) of 84 (80–90) years. Using the Society of Thoracic Surgeons's scoring system for risk factors, there was no significant difference between the groups. Technical success was 84% (n = 16) in Group II. The mean hospital stay in Group II was 20 (2–90) days and the in‐hospital mortality 16% (n = 3). None of these outcomes was significantly different to those in the under 80 group (Group I).

Conclusions

Data from NZTAS registry show outcomes and risk factors do not significantly differ between the two age groups.
Evaluation for fracture patterns around the wrist on three‐dimensional extremity computed tomography, especially focused on the triquetrum
Authors: Oh Eunsun , Kim Hyun‐joo , Hong Seongsook , Hwang Jiyoung , Lim Hyunkyung , Park Sungtae , Hwang Junghwa
Published: December 2014

Introduction

To describe fracture patterns of triquetrum and analyse them according to fracture classifications, anatomy of intrinsic carpal ligaments and comparison with other wrist fractures.

Methods

We retrospectively reviewed 297 three‐dimensional extremity computed tomographies (CTs) on wrist fractures from October 2007 to January 2012. We initially classified the fractures according to the involved bones and analyzed them according to the patterns of triquetrum fractures, associated carpal bone fractures and presence of combined distal radius fractures. We also correlated the fracture patterns with the patient's injury patterns.

Results

A total of 297 CTs and 291 patients were included (162 males and 129 females; mean age, 47.8 years). There were a total of 131 carpal bone fractures in 102 patients of 102 CTs. Triquetrum fractures were the most commonly observed cases (36 cases/27.5%). For the triquetrum fractures, the following types were observed: 26 dorsal, five volar, two comminuted fractures are observed in triquetrum. Three other triquetrum fractures show combined forms of other carpal bone fractures. For the combined distal radius fractures, 10 dorsal and two volar fractures were shown. Out of 187 distal radius fractures, 20 showed carpal bone fractures (10.7%). There were no differences in injury patterns according to the fracture patterns. The most common pattern of injury was falling on the outstretched hand, followed by fall from height.

Conclusion

The dorsum of triquetrum is more frequently fractured than the volar aspect. The number of carpal bone fractures among the patients who have distal radius fractures is higher than usual expectation.
Reverse genetics in the tide pool: knock‐down of target gene expression via RNA interference in the copepod Tigriopus californicus
Authors: Barreto Felipe S. , Schoville Sean D. , Burton Ronald S.
Published: December 2014

Reverse genetic tools are essential for characterizing phenotypes of novel genes and testing functional hypotheses generated from next‐generation sequencing studies. RNA interference (RNAi) has been a widely used technique for describing or quantifying physiological, developmental or behavioural roles of target genes by suppressing their expression. The marine intertidal copepod Tigriopus californicus has become an emerging model for evolutionary and physiological studies, but this species is not amenable to most genetic manipulation approaches. As crustaceans are susceptible to RNAi‐mediated gene knock‐down, we developed a simple method for delivery of gene‐specific double‐stranded RNA that results in significant suppression of target gene transcription levels. The protocol was examined on five genes of interest, and for each, at least 50% knock‐down in expression was achieved. While knock‐down levels did not reach 100% in any trial, a well‐controlled experiment with one heat‐shock gene showed unambiguously that such partial gene suppression may cause dramatic changes in phenotype. Copepods with suppressed expression of heat‐shock protein beta 1 (hspb1) exhibited dramatically decreased tolerance to high temperatures, validating the importance of this gene during thermal stress, as proposed by a previous study. The application of this RNAi protocol in T. californicus will be invaluable for examining the role of genes putatively involved in reproductive isolation, mitochondrial function and local adaptation.
Early prediction of rheumatoid arthritis by magnetic resonance imaging in the absence of anti‐cyclic citrullinated peptide antibodies and radiographic erosions in undifferentiated inflammatory arthritis patients: a prospective study
Authors: Ji Lanlan , Li Guangtao , Xu Yufeng , Zhou Wei , Zhang Zhuoli
Published: December 2014

Objective

To assess the practicability of magnetic resonance imaging (MRI) in confirming the diagnosis of clinically suspected rheumatoid arthritis (RA), when anti‐cyclic citrullinated peptide antibody and radiographic erosions are absent.

Methods

We prospectively involved 31 treatment‐naive patients with early inflammatory arthritis. At the initial visit, X‐rays and gadolinium‐enhanced MRI of both hands, as well as serological examinations and acute phase reactants were performed. The scores of synovitis, bone edema, bone erosion and tenosynovitis of metacarpophalangeal and wrist joints were evaluated using the RA‐MRI scoring system. For all the patients, radiographs at baseline were normal and anti‐cyclic citrullinated peptide antibodies were negative.

Results

At the end of follow‐up(median 15 months, range 12–20 months), 22 patients were diagnosed as having RA according to 1987 American College of Rheumatology criteria. Bone edema, erosions, synovitis and tenosynovitis were observed in all the patients. However, the frequency of symmetric synovitis in wrists was significantly higher in the RA group. Moreover this group turned out to have significantly higher MRI bone erosion score in wrists. Further, receiver operating characteristic curve analysis revealed a positive wrist bone erosion score at 5, with a specificity of 78% and a sensitivity of 68%. There was no significant difference between the two groups with respect to metacarpophalangeal synovitis, metacarpophalangeal bone erosion, bone edema or tenosynovitis.

Conclusion

MRI evidence of symmetric synovitis at wrist and a high bone erosion score at that site may assist in making an early diagnosis of RA in those patients who are negative for anti‐cyclic citrullinated peptide antibody.
Cloninger's psychobiological model of personality and psychological distress in fibromyalgia
Authors: Garcia‐Fontanals Alba , García‐Blanco Susanna , Portell Mariona , Pujol Jesús , Poca‐Dias Violant , García‐Fructuoso Ferran , López‐Ruiz Marina , Gutiérrez‐Rosado Teresa , Gomà‐i‐Freixanet Montserrat , Deus Joan
Published: December 2014

Aim

Personality can play an important role in the clinical symptoms of fibromyalgia (FM). The aim of this study is to identify personality profiles in FM patients and the possible presence of personality disorder (PD) from the Temperament and Character Inventory‐Revised (TCI‐R), and to assess whether personality dimensions are related to psychological distress in FM.

Method

The sample consisted of 42 patients with FM and 38 healthy controls. The TCI‐R, Hospital Anxiety and Depression Scale, State‐Trait Anxiety Inventory, Short‐Form‐36 Health Survey, Fibromyalgia Impact Questionnaire and McGill Pain Questionnaire were administered.

Results

The personality profile of the FM group based on the TCI‐R is defined by high Harm Avoidance (HA), low Novelty Seeking (NS), and low Self‐Directedness (SD). Only one‐third of patients with FM present a possible psychometric PD, principally from Cluster C. In the FM group, HA and SD are associated positively and negatively, respectively, with indicators of emotional distress. Patients with higher HA present higher perceived pain intensity rated via a verbal‐numerical scale while Determination (SD2) reduced the perceived level of pain induced by the stimulus. NS is negatively related to the number of work absences caused by FM.

Conclusions

The study suggests that HA and SD play an important role in psychological distress in FM. The fact that SD is prone to modification and has a regulatory effect on emotional impulses is a key aspect to consider from the psychotherapeutic point of view.
Towards the identification of the loci of adaptive evolution
Authors: Pardo‐Diaz Carolina , Salazar Camilo , Jiggins Chris D.
Published: December 2014

Establishing the genetic and molecular basis underlying adaptive traits is one of the major goals of evolutionary geneticists in order to understand the connection between genotype and phenotype and elucidate the mechanisms of evolutionary change. Despite considerable effort to address this question, there remain relatively few systems in which the genes shaping adaptations have been identified.

Here, we review the experimental tools that have been applied to document the molecular basis underlying evolution in several natural systems, in order to highlight their benefits, limitations and suitability. In most cases, a combination of DNA, RNA and functional methodologies with field experiments will be needed to uncover the genes and mechanisms shaping adaptation in nature.
Reversible cell cycle inhibition and premature aging features imposed by conditional expression of p16Ink4a
Authors: Boquoi Amelie , Arora Sanjeevani , Chen Tina , Litwin Sam , Koh James , Enders Greg H.
Published: December 2014

The cyclin‐dependent kinase (Cdk) inhibitor p16Ink4a (p16) is a canonical mediator of cellular senescence and accumulates in aging tissues, where it constrains proliferation of some progenitor cells. However, whether p16 induction in tissues is sufficient to inhibit cell proliferation, mediate senescence, and/or impose aging features has remained unclear. To address these issues, we generated transgenic mice that permit conditional p16 expression. Broad induction at weaning inhibited proliferation of intestinal transit‐amplifying and Lgr5+ stem cells and rapidly imposed features of aging, including hair loss, skin wrinkling, reduced body weight and subcutaneous fat, an increased myeloid fraction in peripheral blood, poor dentition, and cataracts. Aging features were observed with multiple combinations of p16 transgenes and transactivators and were largely abrogated by a germline Cdk4 R24C mutation, confirming that they reflect Cdk inhibition. Senescence markers were not found, and de‐induction of p16, even after weeks of sustained expression, allowed rapid recovery of intestinal cell proliferation and reversal of aging features in most mice. These results suggest that p16‐mediated inhibition of Cdk activity is sufficient to inhibit cell proliferation and impose aging features in somatic tissues of mammals and that at least some of these aging features are reversible.
Brain function during cognitive flexibility and white matter integrity in alcohol‐dependent patients, problematic drinkers and healthy controls
Authors: Jansen Jochem M. , van Holst Ruth J. , van den Brink Wim , Veltman Dick J. , Caan Matthan W. A. , Goudriaan Anna E.
Published: December 2014

Cognitive flexibility has been associated with prefrontal white matter (WM) integrity in healthy controls (HCs), showing that lower WM integrity is associated with worse performance. Although both cognitive flexibility and WM integrity have been found to be aberrant in alcohol‐dependent (AD) patients, the relationship between the two has never been tested. In this study, we investigated the association between WM tract density and cognitive flexibility in patients with AD (n = 26) and HCs (n = 22). In order to assess the influence of AD severity, we also included a group of problematic drinkers (PrDs; n = 23) who did not meet the AD criteria. Behavioral responses and brain activity during a cognitive flexibility task were measured during functional magnetic resonance imaging. Probabilistic fiber tracking was performed between the dorsolateral prefrontal cortex and the basal ganglia; two crucial regions for task switching. Finally, the task‐related functional connectivity between these areas was assessed. There were no significant group differences in the task performance. However, compared with HCs, AD patients and PrDs showed decreased WM integrity and increased prefrontal brain activation during task switching. Evidence is presented for a compensatory mechanism, involving recruitment of additional prefrontal resources in order to compensate for WM and neural function impairments in AD patients and PrDs. Although present in both alcohol groups, the PrDs were more successful in invoking this compensatory mechanism when compared to the AD patients. We propose that this may therefore serve as a protective factor, precluding transition from problematic drinking into alcohol dependence.
Rat Articular Cartilages Change Their Tissue and Protein Compositions During Perinatal Period
Authors: Kobayashi‐Miura M. , Miura T. , Osago H. , Yamaguchi Y. , Aoyama T. , Tanabe T. , Matsumoto K.‐i. , Fujita Y.
Published: December 2014

Articular cartilage (AC) covers the surface of bones in joints and functions as a cushion against mechanical loading. The tissue contains abundant extracellular matrix (ECM), which mainly consists of proteoglycans (PG) and collagen (COL) fibres. The property of AC is gradually changing by ageing with gravity loading. To know the property change of AC by initial gravity loading during short period after birth, we performed histological assays and proteomics assay on the AC of the femoral condyle in knee joints of perinatal rats. The water content (%) was significantly decreased in neonate AC compared with fetal AC. During the perinatal stages (E19 and P0), the localizations of glycosaminoglycan (GAG) and type I and II COLs were homogeneous. The density of chondrocytes was significantly decreased in the deeper layers comparing with the surface layer in neonate AC. In addition, we found a drastic change in the protein expression pattern on proteomic analysis. The expressions of ECM components were relatively increased in neonate AC compared with fetal AC.
Spindle cell metaplastic carcinoma of breast: A clinicopathological and immunohistochemical analysis
Authors: Zhu Hong , Li Ke , Dong Dan‐dan , Fu Jing , Liu Dan‐dan , Wang Lei , Xu Gang , Song Lin‐hong
Published: December 2014

Aim

To better characterize spindle cell metaplastic carcinoma (SpCMC) of breast, a rare variant of breast cancer that has been classified under the broad rubric of metaplastic carcinoma.

Methods

We presented herein 19 cases of metaplastic breast carcinoma with dominant spindle cell component. All cases were clinically of breast origin, showed more than 80% spindle morphology, 10 cases exhibited pure spindled morphology, 8 contained invasive ductal carcinoma (IDC) and 1 presented with ductal carcinoma in situ elements.

Results

Immunohistochemical studies showed evidence suggesting myoepithelial and epithelial differentiation as exhibited by immunoreactivity for at least one myoepithelial and epithelial markers in all pure spindle cell components. IDC group showed 21.7% of axillary lymph nodes metastasis rate, whereas the axillary lymph node metastasis rate of the SpCMC group was 1.3%, significantly lower than that of the IDC group (P < 0.001). Immunohistochemical staining of IDC exhibited higher degrees of positivity for ER, PR and Her2 (90, 60 and 30%, respectively) when compared with the SpCMC group, which showed a positive degree of 5.2, 5.2 and 10.5% for ER, PR and Her2, respectively (P < 0.001).

Conclusion

Based on this series, SpCMC is a rare variant of metaplastic breast carcinoma with the distinct histopathological and immunohistochemical features. The biological behaviors of SpCMC, like axillary lymph node status, were quite different from that of IDC, suggesting that it may act as an independent pathologic subtype. Immunohistochemical analysis of a panel of epithelial and myoepithelial markers could contribute to the pathologic diagnosis of SpCMC.
Ophthalmic experience over 10 years in an observational nationwide Danish cohort of Fabry patients with access to enzyme replacement
Authors: Fledelius Hans C. , Sandfeld Lisbeth , Rasmussen Åse Krogh , Madsen Christoffer V. , Feldt‐Rasmussen Ulla
Published: December 2014

Purpose

Enzyme replacement therapy (ERT) was offered from year 2001 to patients with Fabry disease. The ophthalmic experience was analysed, as part of a general 10‐year status.

Methods

A retrospective observational series comprising 39 patients (25 females, 14 males) closely followed by the endocrinologists, and with regular ophthalmic control. Time of inclusion was when the option of ERT was started, at age 11–60 years. Eye data (standard eye examination, including retinal imaging) were incomplete in five, due to death or non‐attendance, and five patients had refused treatment.

Results

Vision was normal throughout, except in two young males with total unilateral central retinal artery occlusion, prior to and during enzyme replacement, respectively. Cornea verticillata and conjunctival vessel ectasies were common. Tortuosity of retinal arterioles and venules was recorded in eight and 18 patients, respectively, and phlebopathy in 22, although generally without evidence of loss of vessel wall integrity. Systemic vascular lesions with or without loss of function were recorded for kidney (n = 23), heart (n = 17) and brain (n = 7), and an association was suggested between nephropathy and abnormal morphology of retinal vessels.

Conclusions

Thirteen of 32 patients on ERT showed a reduction of corneal deposits over the study period. Abnormal ocular vessel morphology was a frequent finding. In contrast to the function loss related to systemic ischaemic lesions, we found no indication of impairment of visual parameters in 37. Compared to other Fabry series, two of 39 patients with serious unilateral occlusive retinal disease may appear a high number. The presence of retinal tortuosity is discussed, possibly reflecting haemodynamic events related to vessel wall deposits, but could also be ‘constitutional’, as part of the Fabry inheritance.
Intra‐ocular pressure‐lowering effects of a Rho kinase inhibitor, ripasudil (K‐115), over 24 hours in primary open‐angle glaucoma and ocular hypertension: a randomized, open‐label, crossover study
Authors: Tanihara Hidenobu , Inoue Toshihiro , Yamamoto Tetsuya , Kuwayama Yasuaki , Abe Haruki , Suganami Hideki , Araie Makoto ,   the K‐115 Clinical Study Group
Published: December 2014

Purpose

To investigate the intra‐ocular pressure (IOP)‐lowering effects of a selective Rho kinase inhibitor, ripasudil (K‐115), over 24 hr in patients with primary open‐angle glaucoma (POAG) or ocular hypertension (OHT).

Methods

In this multicenter, prospective, randomized, open‐label, 3‐period, Latin‐square crossover clinical study, 28 patients with POAG or OHT whose IOP level was 21 mmHg or higher were subdivided into three groups. Each patient was treated with placebo and ripasudil in concentrations of 0.2 and 0.4%, at 9:00 and 21:00 on day 1 through a total of 3 periods separated by washout periods. IOP was measured at 9:00, 10:00, 11:00, 13:00, 16:00, 19:00, 21:00, 22:00 and 23:00 on day 1, and 1:00, 4:00, 7:00 and 9:00 on day 2 in sitting position using Goldmann applanation tonometer. Main outcome measure was the IOP reduction of placebo and ripasudil from baseline.

Results

The mean IOP reduction was −5.2 mmHg for 0.2%, −6.4 mmHg for 0.4% and −2.0 mmHg for placebo at 2 hr after the first instillation. Also, the corresponding values were −6.8 mmHg for 0.2%, −7.3 mmHg for 0.4% and −4.1 mmHg for placebo at 2 hr after the second instillation. Statistically significant IOP reduction, compared with placebo, was found for both 0.2 and 0.4% from 1 through 7 hr after each instillation. In safety, conjunctival hyperaemia was observed in 22 patients (79%) for 0.2%, 27 patients (96%) for 0.4% and three patients (11%) for placebo.

Conclusion

Ripasudil is a promising new topical medication to lower IOP for at least 7 hr after instillations in patients with POAG or OHT.
Systemic levels of vascular endothelial growth factor before and after intravitreal injection of aflibercept or ranibizumab in patients with age‐related macular degeneration: a randomised, prospective trial
Authors: Zehetner Claus , Kralinger Martina T. , Modi Yasha S. , Waltl Inga , Ulmer Hanno , Kirchmair Rudolf , Bechrakis Nikolaos E. , Kieselbach Gerhard F.
Published: December 2014

Purpose

To evaluate the changes of vascular endothelial growth factor (VEGF) plasma levels after intravitreal injections of aflibercept or ranibizumab in patients with exudative age‐related macular degeneration (AMD).

Methods

Thirty‐eight patients with exudative AMD were included in this randomised, prospective study. Nineteen patients were randomised to treatment with intravitreal aflibercept (2.0 mg) and 19 to intravitreal ranibizumab (0.5 mg). The concentration of VEGF was measured by ELISA just before the injection, after 7 days and 1 month. Twenty‐two age‐ and sex‐matched healthy patients without chorioretinal diseases served as control.

Results

The median baseline plasma VEGF concentration was 61.0 pg/ml in the control group, 43.0 pg/ml in the aflibercept group and 59.0 pg/ml in the ranibizumab group (p = 0.127). Seven days after intravitreal injection of aflibercept plasma levels were significantly reduced to values below the minimum detectable dose (MDD) in 17 of 19 patients (89.5%) resulting in a median VEGF concentration of <9 pg/ml (p < 0.001). The reduction persisted throughout 1 month with values below the MDD in 5 of 19 patients (26.3%) and a median measurement of 17.0 pg/ml (p < 0.001). In patients treated with ranibizumab no significant effects could be observed with a baseline VEGF of 59.0 pg/ml, 54.0 pg/ml at 7 days (p = 0.776) and 58.5 pg/ml at 4 weeks of follow‐up (p = 0.670).

Conclusion

After intravitreal aflibercept injection, the systemic VEGF levels were significantly reduced throughout the observational period of 4 weeks. No significant systemic effects of intravitreal ranibizumab on plasma VEGF were observed.
A prospective, observational, open‐label, multicentre study to investigate the daily treatment practice of ranibizumab in patients with neovascular age‐related macular degeneration
Authors: van Asten Freekje , Evers‐Birkenkamp Kim U. , van Lith‐Verhoeven Janneke J. C. , de Jong‐Hesse Yvonne , Hoppenreijs Vincent P. T. , Hommersom Richard F. , Scholten Agnes M. , Hoyng Carel B. , Klaver Johannes H. J. ,   the HELIOS study group
Published: December 2014

Purpose

The HELIOS (Health Economics with Lucentis in Observational Settings) study was designed on request of the Dutch Health Authority for an observational study to assess the effectiveness and safety of ranibizumab for neovascular age‐related macular degeneration (wet AMD) in daily practice.

Methods

The HELIOS study was a 2‐year prospective, observational, open‐label, multicentre study involving 14 sites. Patients with wet AMD were enrolled and observed for a period of 24 months. The data were collected at baseline and at the visits closest around the time‐points 3, 6, 12, 18 and 24 months after inclusion.

Results

Treatment with ranibizumab resulted in prevention of vision loss. The mean ETDRS score increased from 45.1 letters at baseline to 48.5 letters at 24 months. This was achieved with a mean of 7.8 injections over 24 months. Stabilization of visual acuity was also reflected by the scores on the quality of life EQ‐5D questionnaire, which did not significantly change over the study period. The more subjective EQ‐VAS questionnaire showed an overall improvement. The VFQ‐25 questionnaire was also mostly stable over time. After 24 months, 32.2% of the patients gained ≥1 letter and 17.1% gained >15 letters. Patients completing the loading phase were better responders, as demonstrated by increased long‐term visual acuity. In addition, ranibizumab was well tolerated and had a safety profile commonly seen in routine clinical practice.

Conclusion

This study demonstrates that also in daily practice ranibizumab was effective in preventing vision loss over a period of 24 months. No new safety findings were identified.
B‐type and N‐terminal pro‐B‐type natriuretic peptides are equally useful in assessing patent ductus arteriosus in very preterm infants
Authors: König Kai , Guy Katelyn J. , Drew Sandra M. , Barfield Charles P.
Published: December 2014

Aim

B‐type natriuretic peptide (BNP) and N‐terminal pro‐BNP (NTproBNP) have been shown to correlate with the size of the patent ductus arteriosus (PDA) in preterm infants. We investigated whether BNP or NTproBNP was more closely correlated with PDA size.

Methods

This prospective observational study included preterm infants born <32 weeks' gestation who had an echocardiogram performed within the first four days of life. Blood samples were taken simultaneously for BNP and NTproBNP measurements prior to echocardiographic examination.

Results

Of the 60 infants recruited, 58 had complete data sets. The cohort's mean and standard deviation (SD) gestational age was 27+3 (2+2) weeks, the mean (SD) birthweight was 1032 (315) grams, and 46 (79.3%) infants had a PDA with a mean (SD) diameter of 3.2 (0.9) mm. Median and interquartile range (IQR) BNP levels were 486.5 (219–1316) pg/mL for infants with a PDA and 190 (95.5–514.5) pg/mL for infants without a PDA. Median (IQR) NTproBNP levels were 10 858.5 (6319–42 108) pg/mL for infants with a PDA and 7488 (3363–14 227.5) pg/mL for infants without a PDA. Both BNP (R = 0.35, p = 0.0066) and NTproBNP (R = 0.31, p = 0.018) were significantly correlated with PDA size.

Conclusion

BNP and NTproBNP are similarly useful for assessing PDA size in preterm infants.
Neuropeptide Y in the noradrenergic neurones induces obesity and inhibits sympathetic tone in mice
Authors: Vähätalo L. H. , Ruohonen S. T. , Mäkelä S. , Kovalainen M. , Huotari A. , Mäkelä K. A. , Määttä J. A. , Miinalainen I. , Gilsbach R. , Hein L. , Ailanen L. , Mattila M. , Eerola K. , Röyttä M. , Ruohonen S. , Herzig K. ‐H. , Savontaus E.
Published: December 2014

Aim

Neuropeptide Y (NPY) co‐localized with noradrenaline in central and sympathetic nervous systems seems to play a role in the control of energy metabolism. In this study, the aim was to elucidate the effects and pathophysiological mechanisms of increased NPY in catecholaminergic neurones on accumulation of body adiposity.

Methods

Transgenic mice overexpressing NPY under the dopamine‐beta‐hydroxylase promoter (OE‐NPYDβH) and wild‐type control mice were followed for body weight gain and body fat content. Food intake, energy expenditure, physical activity, body temperature, serum lipid content and markers of glucose homoeostasis were monitored. Thermogenic and lipolytic responses in adipose tissues, and urine catecholamine and tissue catecholamine synthesizing enzyme levels were analysed as indices of sympathetic tone.

Results

Homozygous OE‐NPYDβH mice showed significant obesity accompanied with impaired glucose tolerance and insulin resistance. Increased adiposity was explained by neither increased food intake or fat absorption nor by decreased total energy expenditure or physical activity. Adipocyte hypertrophy and decreased circulating lipid levels suggested decreased lipolysis and increased lipid uptake. Brown adipose tissue thermogenic capacity was decreased and brown adipocytes filled with lipids. Enhanced response to adrenergic stimuli, downregulation of catecholamine synthesizing enzyme expressions in the brainstem and lower adrenaline excretion supported the notion of low basal catecholaminergic activity.

Conclusion

Increased NPY in catecholaminergic neurones induces obesity that seems to be a result of preferential fat storage. These results support the role of NPY as a direct effector in peripheral tissues and an inhibitor of sympathetic activity in the pathogenesis of obesity.