bluuebunny/crossref_metadata_2025 huggingface dataset:
Pulmonary Hypertension in Lambs Transfused with Stored Blood Is Prevented by Breathing Nitric Oxide
Authors: Baron David M. , Yu Binglan , Lei Chong , Bagchi Aranya , Beloiartsev Arkadi , Stowell Christopher P. , Steinbicker Andrea U. , Malhotra Rajeev , Bloch Kenneth D. , Zapol Warren M.
Published: January 2012

Background

During extended storage, erythrocytes undergo functional changes. These changes reduce the viability of erythrocytes leading to release of oxyhemoglobin, a potent scavenger of nitric oxide. We hypothesized that transfusion of ovine packed erythrocytes (PRBC) stored for prolonged periods would induce pulmonary vasoconstriction in lambs, and that reduced vascular nitric oxide concentrations would increase this vasoconstrictor effect.

Methods

We developed a model of autologous stored blood transfusion in lambs (n = 36). Leukoreduced blood was stored for either 2 days (fresh PRBC) or 40 days (stored PRBC). Fresh or stored PRBC were transfused into donors instrumented for awake hemodynamic measurements. Hemodynamic effects of PRBC transfusion were also studied after infusion of N-nitro-L-arginine methyl-ester (25 mg/kg) or during inhalation of nitric oxide (80 ppm).

Results

Cell-free hemoglobin levels were higher in the supernatant of stored PRBC than in supernatant of fresh PRBC (Mean ± SD, 148 ± 20 vs. 41 ± 13 mg/dl, respectively, P < 0.001). Pulmonary artery pressure during transfusion of stored PRBC transiently increased from 13 ± 1 to 18 ± 1 mmHg (P < 0.001) and was associated with increased plasma hemoglobin concentrations. N-nitro-L-arginine methyl-ester potentiated the increase in pulmonary arterial pressure induced by transfusing stored PRBC, whereas inhalation of nitric oxide prevented the vasoconstrictor response.

Conclusions

Our results suggest that patients with reduced vascular nitric oxide levels because of endothelial dysfunction may be more susceptible to adverse effects of transfusing blood stored for prolonged periods. These patients might benefit from transfusion of fresh PRBC, when available, or inhaled nitric oxide supplementation to prevent the pulmonary hypertension associated with transfusion of stored PRBC.
Choice of Anesthetic Combination Determines Ca2+Leak after Ischemia-Reperfusion Injury in the Working Rat Heart
Authors: Zaugg Michael , Wang Lianguo , Zhang Liyan , Lou Phing-How , Lucchinetti Eliana , Clanachan Alexander S.
Published: January 2012

Background

There is a lack of studies investigating cardioprotection by common combinations of anesthetics. However, because a general anesthetic consists of a mixture of drugs with potentially interfering effects on signaling and cytoprotection, the most favorable combination should be used.

Methods

Working rat hearts were exposed to 20 min of ischemia and 30 min of reperfusion. Periischemic sevoflurane (2 vol-%), propofol (10 μM), or remifentanil (3 nM) (single treatments) and the three combinations thereof (combination treatments) were assessed for their ability to improve postischemic left ventricular work and to prevent intracellular Ca leak and overload. Beat-to-beat oscillations in intracellular [Ca] were measured using indo-1 AM. Phosphorylation of calcium/calmodulin-dependent protein kinase IIδ, ryanodine receptor-2, and phospholamban was determined.

Results

The single treatments with sevoflurane or remifentanil were highly protective with respect to functional recovery and Ca overload, but propofol, even at high concentrations, did not show similar protection. Sevoflurane combined with propofol completely lost its protection in the presence of low sedative propofol concentrations (≥1 μM), whereas remifentanil combined with propofol (10 μM) retained its protection. Propofol antagonism of sevoflurane protection was concentration-dependent and mimicked by the reactive oxygen species scavenger N-2-mercaptopropionyl-glycine. Addition of propofol to sevoflurane activated calcium/calmodulin-dependent protein kinase type IIδ and hyperphosphorylated the ryanodine receptor-2, consistent with causing a postischemic Ca leak from the sarcoplasmic reticulum. Remifentanil did not enhance sevoflurane protection.

Conclusions

The choice of anesthetic combination determines the postischemic Ca leak and intracellular overload. The results from these experiments will help to design studies for optimizing perioperative cardioprotection in high-risk surgical patients.
Continuous Femoral Nerve Blocks
Authors: Bauer Maria , Wang Lu , Onibonoje Olusegun K. , Parrett Chad , Sessler Daniel I. , Mounir-Soliman Loran , Zaky Sherif , Krebs Viktor , Buller Leonard T. , Donohue Michael C. , Stevens-Lapsley Jennifer E. , Ilfeld Brian M.
Published: January 2012

Background

Whether decreasing the local anesthetic concentration during a continuous femoral nerve block results in less quadriceps weakness remains unknown.

Methods

Preoperatively, bilateral femoral perineural catheters were inserted in subjects undergoing bilateral knee arthroplasty (n = 36) at a single clinical center. Postoperatively, right-sided catheters were randomly assigned to receive perineural ropivacaine of either 0.1% (basal 12 ml/h; bolus 4 ml) or 0.4% (basal 3 ml/h; bolus 1 ml), with the left catheter receiving the alternative concentration/rate in an observer- and subject-masked fashion. The primary endpoint was the maximum voluntary isometric contraction of the quadriceps femoris muscles the morning of postoperative day 2. Equivalence of treatments would be concluded if the 95% CI for the difference fell within the interval -20%-20%. Secondary endpoints included active knee extension, passive knee flexion, tolerance to cutaneous electrical current applied over the distal quadriceps tendon, dynamic pain scores, opioid requirements, and ropivacaine consumption.

Results

Quadriceps maximum voluntary isometric contraction for limbs receiving 0.1% ropivacaine was a mean (SD) of 13 (8) N · m, versus 12 (8) N · m for limbs receiving 0.4% [intrasubject difference of 3 (40) percentage points; 95% CI -10-17; P = 0.63]. Because the 95% CI fell within prespecified tolerances, we conclude that the effect of the two concentrations were equivalent. Similarly, there were no statistically significant differences in secondary endpoints.

Conclusions

For continuous femoral nerve blocks, we found no evidence that local anesthetic concentration and volume influence block characteristics, suggesting that local anesthetic dose (mass) is the primary determinant of perineural infusion effects.
Exceptionally Stable Fluorous Emulsions for the Intravenous Delivery of Volatile General Anesthetics
Authors: Jee Jun-Pil , Parlato Maria C. , Perkins Mark G. , Mecozzi Sandro , Pearce Robert A.
Published: January 2012

Background

IV delivery of volatile fluorinated anesthetics has a number of potential advantages when compared with the current inhalation method of administration. We reported previously that the IV delivery of sevoflurane can be achieved through an emulsion composed of a linear fluorinated diblock copolymer, a stabilizer, and the anesthetic. However, this original emulsion was subject to particle size growth that would limit its potential clinical utility. We hypothesized that the use of bulkier fluorous groups and smaller polyethylene glycol moieties in the polymer design would result in improved emulsion stability while maintaining anesthetic functionality.

Methods

The authors prepared emulsions incorporating sevoflurane, perfluorooctyl bromide as a stabilizing agent, and combinations of linear fluorinated diblock copolymer and a novel dibranched fluorinated diblock copolymer. Emulsion stability was assessed using dynamic light scattering. The ability of the emulsions to induce anesthesia was tested in vivo by administering them intravenously to 15 male Sprague-Dawley rats and measuring loss of the forepaw righting reflex.

Results

20% (volume/volume) sevoflurane emulsions incorporating mixtures of dibranched and linear diblock copolymers had improved stability, with those containing an excess of the dibranched polymers displaying stability of particle size for more than 1 yr. The ED50s for loss of forepaw-righting reflex were all similar, and ranged between 0.55- 0.60 ml/kg body weight.

Conclusions

Hemifluorinated dibranched polymers can be used to generate exceptionally stable sevoflurane nanoemulsions, as required of formulations intended for clinical use. IV delivery of the emulsion in rats resulted in induction of anesthesia with rapid onset and smooth and rapid recovery.
Optimal decision-making in mammals: insights from a robot study of rodent texture discrimination
Authors: Lepora Nathan F. , Fox Charles W. , Evans Mathew H. , Diamond Mathew E. , Gurney Kevin , Prescott Tony J.
Published: January 2012

Texture perception is studied here in a physical model of the rat whisker system consisting of a robot equipped with a biomimetic vibrissal sensor. Investigations of whisker motion in rodents have led to several explanations for texture discrimination, such as resonance or stick-slips. Meanwhile, electrophysiological studies of decision-making in monkeys have suggested a neural mechanism of evidence accumulation to threshold for competing percepts, described by a probabilistic model of Bayesian sequential analysis. For our robot whisker data, we find that variable reaction-time decision-making with sequential analysis performs better than the fixed response-time maximum-likelihood estimation. These probabilistic classifiers also use whatever available features of the whisker signals aid the discrimination, giving improved performance over a single-feature strategy, such as matching the peak power spectra of whisker vibrations. These results cast new light on how the various proposals for texture discrimination in rodents depend on the whisker contact mechanics and suggest the possibility of a common account of decision-making across mammalian species.
Cell-surface expression of PrPC and the presence of scrapie prions in the blood of goats
Authors: Dassanayake Rohana P. , Schneider David A. , Herrmann-Hoesing Lynn M. , Truscott Thomas C. , Davis William C. , O’Rourke Katherine I.
Published: January 2012

Although host-encoded prion protein (PrPC) expression in ovine PBMCs and prion infectivity in scrapie-infected sheep blood have been demonstrated, such studies have not been reported in goats. Therefore, this study characterized cell-surface expression of PrPC on PBMC subsets derived from normal goats and sheep, by flow cytometry, and determined prion infectivity in blood from a scrapie-infected goat using a transfusion bioassay in goat kids. Cell-surface PrPC expression was detected on all subsets of goat PBMCs. The highest PrPC cell-surface expression was found in CD2+ T lymphocytes in goats. Transmission of infection was detected in all three recipients who received whole blood from a goat with classical scrapie. It was concluded that caprine PBMCs express PrPC similarly to sheep but with relative differences among PBMCs subsets, and that blood-borne infectious prions can be detected in scrapie-infected goats. Thus, similar to sheep, goat blood may be a suitable diagnostic target for the detection of scrapie infection.
Encapsidation of DNA, a protein and a fluorophore into virus-like particles by the capsid protein of cucumber mosaic virus
Authors: Lu Xiaoyun , Thompson Jeremy R. , Perry Keith L.
Published: January 2012

An important property of some spherical plant viruses is their ability to reassemble in vitro from native capsid protein (CP) and RNA into infectious virus-like particles (VLPs). Virions of cucumber mosaic virus (CMV) are stabilized by protein–RNA interactions and the nucleic acid is essential for assembly. This study demonstrated that VLPs will form in the presence of both ssDNA and dsDNA oligonucleotides, and with a lower size limit of 20 nt. Based on urea disruption assays, assembled VLPs from CMV CP and RNA (termed ReCMV) exhibited a level of stability similar to that of virions purified from plants, whilst VLPs from CMV CP and a 20mer exhibited comparable or greater stability. Fluorescent labelling of VLPs was achieved by the encapsidation of an Alexa Fluor 488-labelled 45mer oligonucleotide (ReCMV-Alexa488-45) and confirmed by transmission electron and confocal microscopy. Using ssDNA as a nucleating factor, encapsidation of fluorescently labelled streptavidin (53 kDa) conjugated to a biotinylated oligonucleotide was observed. The biological activity and stability of ReCMV and ReCMV-Alexa488-45 was confirmed in infectivity assays and insect vector feeding assays. This work demonstrates the utility of CMV CP as a protein cage for use in the growing repertoire of nanotechnological applications.
Crystallization and preliminary X-ray diffraction analysis of orotate phosphoribosyltransferase from the human malaria parasitePlasmodium falciparum
Authors: Takashima Yasuhide , Mizohata Eiichi , Tokuoka Keiji , Krungkrai Sudaratana R. , Kusakari Yukiko , Konishi Saki , Satoh Atsuko , Matsumura Hiroyoshi , Krungkrai Jerapan , Horii Toshihiro , Inoue Tsuyoshi
Published: January 2012

Orotate phosphoribosyltransferase (OPRT) catalyzes the Mg2+-dependent condensation of orotic acid (OA) with 5-α-D-phosphorylribose 1-diphosphate (PRPP) to yield diphosphate (PPi) and the nucleotide orotidine 5′-monophosphate. OPRT fromPlasmodium falciparumproduced inEscherichia coliwas crystallized by the sitting-drop vapour-diffusion method in complex with OA and PRPP in the presence of Mg2+. The crystal exhibited tetragonal symmetry, belonging to space groupP41orP43, with unit-cell parametersa=b= 49.15,c = 226.94 Å. X-ray diffraction data were collected to 2.5 Å resolution at 100 K using a synchrotron-radiation source.
Purification, crystallization and preliminary X-ray crystallographic studies of theMycobacterium tuberculosisDNA gyrase CTD
Authors: Darmon Amélie , Piton Jérémie , Roué Mélanie , Petrella Stéphanie , Aubry Alexandra , Mayer Claudine
Published: January 2012

Mycobacterium tuberculosisDNA gyrase, a nanomachine involved in regulation of DNA topology, is the only type II topoisomerase present in this organism and hence is the sole target of fluoroquinolone in the treatment of tuberculosis. The C-terminal domain (CTD) of the DNA gyrase A subunit possesses a unique feature, the ability to wrap DNA in a chiral manner, that plays an essential role during the catalytic cycle. A construct of 36 kDa corresponding to this domain has been overproduced, purified and crystallized. Diffraction data were collected to 1.55 Å resolution. Cleavage of the N-terminal His tag was crucial for obtaining crystals. The crystals belonged to space groupP212121, with one molecule in the asymmetric unit and a low solvent content (33%). This is the first report of the crystallization and preliminary X-ray diffraction studies of a DNA gyrase CTD from a species that contains one unique type II topoisomerase.
Crystallization and X-ray diffraction analysis of chondroitin lyase from baculovirus: envelope protein ODV-E66
Authors: Kawaguchi Yoshirou , Sugiura Nobuo , Onishi Momo , Kimata Koji , Kimura Makoto , Kakuta Yoshimitu
Published: January 2012

Baculovirus envelope protein ODV-E66 (67–704), in which the N-terminal 66 amino acids are truncated, is a chondroitin lyase. It digests chondroitin and chondroitin 6-sulfate efficiently, but does not digest chondroitin 4-sulfate. This unique characteristic is useful for the preparation of specific chondroitin oligosaccharides and for investigation of the mechanism of baculovirus infection. ODV-E66 (67–704) was crystallized; the crystal diffracted to 1.8 Å resolution and belonged to space groupP62orP64, with unit-cell parametersa = b = 113.5,c= 101.5 Å. One molecule is assumed to be present per asymmetric unit, which gives a Matthews coefficient of 2.54 Å3 Da−1.
Crystallization and preliminary X-ray diffraction analysis of human dipeptidyl peptidase 10 (DPPY), a component of voltage-gated potassium channels
Authors: Bezerra Gustavo Arruda , Dobrovetsky Elena , Seitova Alma , Dhe-Paganon Sirano , Gruber Karl
Published: January 2012

Dipeptidyl peptidase 10 (DPP10, DPPY) is an inactive peptidase associated with voltage-gated potassium channels, acting as a modulator of their electrophysiological properties, cell-surface expression and subcellular localization. Because potassium channels are important disease targets, biochemical and structural characterization of their interaction partners was sought. DPP10 was cloned and expressed using an insect-cell system and the protein was purifiedvia His-tag affinity and size-exclusion chromatography. Crystals obtained by the sitting-drop method were orthorhombic, belonging to space groupP212121with unit-cell parametersa= 80.91,b= 143.73,c= 176.25 Å. A single solution with two molecules in the asymmetric unit was found using the structure of DPP6 (also called DPPX; PDB entry 1xfd) as the search model in a molecular replacement protocol.
Exploring the obesity‐asthma link: do all types of adiposity increase the risk of asthma?
Authors: Fenger R. V. , Gonzalez‐Quintela A. , Vidal C. , Gude F. , Husemoen L. L. , Aadahl M. , Berg N. D. , Linneberg A.
Published: January 2012

Background

Obesity and risk of asthma are linked. Different distributions of adiposity, such as visceral, subcutaneous or ectopic adiposity, may affect asthma risk differently.

Objective

To explore the association of different adiposity types with self‐reported asthma, bronchial inflammation and lung function, accounting for possible effect modifiers, such as atopy and gender.

Methods

In a general population sample of 3471 persons aged 19–72, visceral adipose tissue (VAT) and subcutaneous adipose tissue (SAT) were measured by ultrasound, and fat percentage by bio‐impedance. Body mass index, waist circumference, waist‐to‐hip ratio (WHR), bronchial inflammation as fractional expiratory nitric oxide (
FeNO
), lung function [FEV
1 and forced vital capacity (FVC)], and atopy (specific IgE) were measured.

Results

All adiposity measures were associated with a higher risk of asthma. The risk estimates (odds ratios, OR, with 95% confidence interval, CI) of current asthma were of similar magnitude for all six adiposity measures ranging between 1.17, CI = 0.98–1.40 (SAT) and 1.51, CI = 1.17–1.95 (WHR). The adiposity‐asthma associations were significantly stronger in non‐atopics than in atopics. In non‐atopics the risk estimates of current asthma ranged between 1.35 CI = 1.08–1.72 and 1.82 CI = 1.34–2.46 for SAT and WHR respectively. Consistent results were obtained using dichothomized adiposity measures (obese vs. non‐obsese). The FVC and FEV
1 decreased significantly with increasing adiposity in both atopics and non‐atopics, e.g. FVC decreased between 36 mL (CI = 10, 62 mL) and 155 mL (CI = 124, 186 mL) for one unit (standard error) increase of SAT and VAT respectively. Adiposity measures were not associated with atopy and not consistently associated with
FeNO
levels.

Conclusions and Clinical Relevance

The effect of adiposity on asthma was mainly seen in non‐atopics and did not appear to depend on the distribution of adiposity as reflected by the adiposity measures used in the present study. Increasing adiposity was associated with lower lung function independent of atopic status.
Non‐native fish impacts on Mediterranean freshwater ecosystems: current knowledge and research needs
Authors: RIBEIRO F. , LEUNDA P. M.
Published: January 2012

Abstract  Non‐native fishes are often blamed for exerting adverse ecological impacts on native fishes, but the evidence is circumstantial and the true impacts remain largely unknown. This dilemma is particularly acute for freshwater ecosystems of the Mediterranean Region, where a high level of endemism (a biodiversity hotspot) makes native fish extinctions much more likely because of their small natural ranges. The aim of this paper is to review non‐native fish impacts, from genes to ecosystem level, in the fresh waters of the Mediterranean Region and identify gaps in knowledge that currently hamper effective conservation and management of native species and ecosystems. Most studies reviewed reported potential impacts rather than realised impact mechanisms, and these were conducted predominantly in the European part of the Mediterranean Region. No studies of realised impacts were found for three of the 15 most introduced fishes, which emphasises the need for evidence‐based studies of non‐native species impacts.
Utility of direct observational methods for assessing competitive interactions between non‐native and native freshwater fishes
Authors: ALMEIDA D. , GROSSMAN G. D.
Published: January 2012

Abstract  Biological invasions are a major threat to global freshwater biodiversity. Competition is a frequent negative interaction between non‐native and native species, and this process is commonly quantified using indirect methods (e.g. dietary overlap, comparison of habitat use/selection). However, direct observation can provide crucial information on these biotic interactions. This paper reviews studies that used direct observational methods to quantify interactions between non‐native and native freshwater fishes. Although laboratory and field studies using direct observations are not common, both have been used to demonstrate impacts by non‐native species on native species. These effects include: (1) altered habitat selection and foraging behaviour and (2) disruption of reproductive behaviour. Direct observational techniques have great potential for quantifying the impacts of non‐natives on native freshwater fishes and can detect negative behavioural impacts that would be missed using indirect methods.
Elevated haemocyte number is associated with infection and low fitness potential in wild Daphnia magna
Authors: Auld Stuart K. J. R. , Graham Andrea L. , Wilson Philip J. , Little Tom J.
Published: January 2012

1. Immune activity may be a cause of resistance to parasites, but it can also be a consequence of infection. Thus, the adaptive significance of an immune response is more accurately assessed when it is measured alongside both host fitness and infection status.

2. We sought to determine the significance of immune responses in a naturally coevolving host–parasite system in the wild, with support from laboratory experiments. We measured haemocyte numbers in Daphnia magna in relation to an infection that has a clear fitness consequence: infection with the bacterium Pasteuria ramosa causes sterilization.

3. Haemocyte number was consistently elevated in infected Daphnia in the field and in parasite exposed or infected hosts in the laboratory. Thus, elevated haemocyte numbers were essentially a symptom of infection, and we found no evidence that haemocytes help hosts exclude the parasite.

4. Consequently, these results provide an especially clear example where increased immune activity does not mean increased immunity or fitness: hosts with the highest haemocyte counts have extremely low health and low fitness potential.
Zone of main contribution to the measured signal for a circular current loop source and receiver on the surface of a conductive half‐space
Authors: Machado Marcos Vitor Barbosa , Dias Carlos Alberto
Published: January 2012

The electric and magnetic fields generated by an individual horizontal current ring induced inside a homogeneous conductive half‐space, originating from an external large circular loop source of current in the presence of a flat half‐space, are deduced. A check of self‐consistency for these expressions led to the known general functions for these fields due to the same external source in the presence of that medium. The current rings’ mutual coupling related to the magnetic field's radial component is thoroughly analysed and its specific members are presented. The existence of a relatively small zone inside the half‐space responsible for the main contribution for the signal measured at the observation point, with the source and receiver on the ground surface, is made evident. For increasing values of frequency, at a given transmitter‐receiver (T–R) configuration, this zone shrinks and its central point moves away from a maximum depth of about 30% and horizontal distance of nearly 85%, of the T–R separation, to a point very close to the receiver position. The coordinates of the central point of this zone of main contribution are provided as approximated functions in terms of the induction number .
Characterisation and validation of a novel panel of the six short tandem repeats for genetic counselling in Chinese haemophilia A pedigrees
Authors: DING Q. L. , LU Y. L. , DAI J. , XI X. D. , WANG X. F. , WANG H. L.
Published: January 2012

Summary.  Haemophilia A (HA) is the most common hereditary bleeding disorder caused by F8 gene mutation. Linkage analysis is an auxiliary strategy to direct mutation analysis for genetic counselling of HA. Here we characterize and validate a novel panel of six short tandem repeat (STR) loci for genetic counselling in Chinese HA pedigrees. The panel was analysed in 116 unrelated healthy female patients and 108 male patients, and verified in 169 unrelated pedigrees with HA. The six STR loci in the panel spanned a distance of 0.3 Mb from each side of the F8 gene. Three of them, F8Up226, F8Up146 and F8Down48, were first described here. Markers F8Up226, F8Up146, F8Int13, F8Int25, F8Down48 and DXS1073 exhibited the number of alleles 16, 9, 8, 6, 9 and 10, and heterozygosity rates of 74.8%, 44.8%, 60.9%, 42.6%, 61.7% and 62.0% respectively. Haplotype frequencies analysis suggested that the genotypes of haplotype provided a highly informative content (56.5%). The panel was informative in 167 of 169 unrelated haemophilic pedigrees with the combined diagnostic rate of 98.8%. In eight pedigrees could not be diagnosed by mutation detection linkage studies using the panel were informative in all the pedigrees and a reliable diagnosis was made in seven pedigrees. The novel panel of the six STR loci represents a high degree of informativeness and a low fraction of recombination. Linkage analysis using this panel provides an alternative strategy when direct mutation detection is not feasible for genetic counselling in Chinese HA families.
High Wilms’ tumour gene (WT1) expression and low mitotic count are independent predictors of survival in diffuse peritoneal mesothelioma
Authors: Scattone Anna , Serio Gabriella , Marzullo Andrea , Nazzaro Pietro , Corsi Fabrizio , Cocca Maria Pia , Mattoni Marilena , Punzi Alessandra , Gentile Mattia , Buonadonna Antonia Lucia , Pennella Antonio
Published: January 2012

Scattone A, Serio G, Marzullo A, Nazzaro P, Corsi F, Cocca M P, Mattoni M, Punzi A, Gentile M, Buonadonna A L & Pennella A  (2012) Histopathology60, 472–481  High Wilms’ tumour gene (WT1) expression and low mitotic count are independent predictors of survival in diffuse peritoneal mesothelioma

Aims:  To evaluate the use of the Wilms’ tumour gene (WT1) marker and histomorphological parameters as indicators of prognosis in malignant peritoneal mesothelioma (MPM).

Methods and results:  Histological samples of 31 MPM were stained immunohistochemically for the WT1 protein. The results were quantified by recording the number of stained nuclei, and then correlated with patient survival. Statistical correlation was evaluated for tumour histotype, mitotic count (MC), nuclear grade (NG), necrosis, lymphoid response (grade of inflammation) and desmoplasia with regard to survival. High‐grade histology (solid epithelioid, pure sarcomatoid or biphasic tumours), high NG, MC more than five per 10 per high‐power field (HPF), necrosis and desmoplasia were associated with a significantly worse prognosis. Patients with MPM with low WT1 expression (≤25% of positive cells) survived for a significantly shorter time compared to those with high WT1 expression (>25% of positive cells) (P = 0.0001). The 50% survival time of subjects with low WT1 expression was 2.9 months [95% confidence interval (CI): 2.05–3.71] versus 31.5 months (95% CI: 20.4–42.5) for those with high WT1 expression. On multivariate analysis, WT1 and MC were found to be associated independently with survival (P = 0.002; P = 0.005, respectively).

Conclusions:  Our study suggests that low WT1 expression and high MC may be indicative of an unfavourable prognosis in patients with advanced malignant peritoneal mesothelioma.
Evaluation of table olive by‐product as a source of natural antioxidants
Authors: González‐Hidalgo Inés , Bañón Sancho , Ros José María
Published: January 2012

The main by‐product from the table olive canning industry is the stone with some residual olive flesh. The purpose of this study was to evaluate the composition – phenolic compounds (hydroxytyrosol, tyrosol and oleuropein) and tocopherol – and the antioxidant activity in different fractions (flesh, stone and seed) from the table olive by‐product and the whole by‐product. The highest amounts of phenolic compounds (1710.0 ± 33.8 mg kg−1) as well as the highest antioxidant activity (8226.9 ± 9.9 hydroxytyrosol equivalents mg kg−1) were obtained in the seed. The highest amounts of hydroxytyrosol (854.8 ± 66.0 mg kg−1) and tyrosol (423.6 ± 56.9 mg kg−1) were found in the whole by‐product from the pepper stuffed olives, while the stone without seed had the maximum oleuropein content (750.2 ± 85.3 mg kg−1). α‐Tocopherol values were between 79.8 ± 20.8 mg kg−1 in the seed of the olive stone and 6.2 ± 1.2 mg kg−1 in the whole by‐product from the anchovy‐stuffed olives. In light of the results obtained, it would seem possible to use table olive by‐product as a source of natural antioxidants in foods, cosmetics or pharmaceutical products, thus contributing to diminishing the environmental impact of table olive by‐product and to its revalorisation.
The effect of the high pressure homogenisation on the activity and stability of a commercial neutral protease from Bacillus subtilis
Authors: Tribst Alline A. L. , Augusto Pedro E. D. , Cristianini Marcelo
Published: January 2012

The activity of a commercial neutral protease from Bacillus subtilis after high pressure homogenisation (HPH) was investigated. The enzyme was processed up to 2000 bar, and the residual activity was measured from 20 to 70 °C during refrigerated storage. Moreover, the effect of HPH at high temperatures was evaluated. No improvement in the activities at 55–70 °C were observed after HPH, while an increase of approximately 30% in the 20 °C‐activity was reached after 2000 bar processing. Thus, HPH shifted the optimum temperature from 55 to 20 °C. The high temperature homogenisation caused no changes in 55 °C‐activity, but reduced 20 °C‐activity three times. It suggests that HPH modifies the protease configuration, changing enzyme performance (maximum activity condition), as the efficacy of lock‐and‐key mechanism is strictly dependent on enzyme spatial structure. The changes can be permanent or not, depending on homogenisation pressure, inlet temperature and enzyme storage conditions. Therefore, the HPH is a promising method to change protease characteristics.
Impact of caries onset on number and distribution of new lesions in preschool children
Authors: LEROY ROOS , DECLERCK DOMINIQUE
Published: January 2012

International Journal of Paediatric Dentistry 2013; 23: 39–47

Background.  Caries in preschool children remains an important public health issue.

Aim.  To determine (i) which teeth and tooth surfaces are most susceptible to dental caries by age 3, (ii) where do caries lesions develop during 2‐year follow‐up, and (iii) to evaluate the impact of caries onset on the distribution of new caries experience.

Design.  One thousand and fifty seven consecutively born children were recruited in Flanders (Belgium). Parents completed validated questionnaires on oral health‐related behaviour and trained dentists examined the children at ages 3 and 5.

Results.  Children with visible caries experience at age 3 were significantly more vulnerable in developing additional caries during follow‐up. In this group, new caries experience developed primarily in the occlusal and distal surfaces of the mandibular first molars and the occlusal surfaces of the maxillary second and first molars, whereas in the caries‐free group, the occlusal surfaces of both mandibular and maxillary second molars ranked first.

Conclusions.  This paper confirms the higher vulnerability for further caries development in those children with caries experience at age 3. Visible caries develops most frequently in the occlusal surfaces of the second molars: in high‐risk children already by age 3 and in children who were caries free at baseline by age 5.
Complexity of multitrophic interactions in a grassland ecosystem depends on plant species diversity
Authors: Rzanny Michael , Voigt Winfried
Published: January 2012

1. We studied the theoretical prediction that a loss of plant species richness has a strong impact on community interactions among all trophic levels and tested whether decreased plant species diversity results in a less complex structure and reduced interactions in ecological networks.

2. Using plant species‐specific biomass and arthropod abundance data from experimental grassland plots (Jena Experiment), we constructed multitrophic functional group interaction webs to compare communities based on 4 and 16 plant species. 427 insect and spider species were classified into 13 functional groups. These functional groups represent the nodes of ecological networks. Direct and indirect interactions among them were assessed using partial Mantel tests. Interaction web complexity was quantified using three measures of network structure: connectance, interaction diversity and interaction strength.

3. Compared with high plant diversity plots, interaction webs based on low plant diversity plots showed reduced complexity in terms of total connectance, interaction diversity and mean interaction strength. Plant diversity effects obviously cascade up the food web and modify interactions across all trophic levels. The strongest effects occurred in interactions between adjacent trophic levels (i.e. predominantly trophic interactions), while significant interactions among plant and carnivore functional groups, as well as horizontal interactions (i.e. interactions between functional groups of the same trophic level), showed rather inconsistent responses and were generally rarer.

4. Reduced interaction diversity has the potential to decrease and destabilize ecosystem processes. Therefore, we conclude that the loss of basal producer species leads to more simple structured, less and more loosely connected species assemblages, which in turn are very likely to decrease ecosystem functioning, community robustness and tolerance to disturbance. Our results suggest that the functioning of the entire ecological community is critically linked to the diversity of its component plants species.
A descriptive study of the experiences of lesbian, gay and transgender parents accessing health services for their children
Authors: Chapman Rose , Wardrop Joan , Freeman Phoenix , Zappia Tess , Watkins Rochelle , Shields Linda
Published: January 2012

Aim. To explore the experiences of lesbian, gay and transgender families accessing health care for their children.

Background. Although lesbian, gay and transgender families are becoming more common, little is known about their health‐seeking experiences. These families may be fearful about disclosing their sexual orientation or gender identity to health professionals. As a result, lesbian, gay and transgender parents may not be receiving optimal care for their children.

Design. Descriptive qualitative study.

Method. Data were collected through semi‐structured interviews with 11 lesbian, gay and transgender parents in Australia.

Results. Three themes were generated from the data: ‘managing health care experiences’, ‘attitudes’ and ‘transforming bureaucracies’. Negative experiences included encountering homophobia or transphobia and being required to educate health professionals. Positive experiences occurred when both parents were acknowledged as having an equal say in their child’s health care.

Conclusion. Many health professionals lack the skill or knowledge to meet the needs of lesbian, gay and transgender families. Health services are required to ensure that all policies and procedures are inclusive of all family constellations and that staff receive relevant and up‐to‐date sensitivity training and create an environment that is respectful of all family groups.

Relevance to clinical practice. Adopting a philosophy of family centred care can enable health providers and health professionals to provide lesbian, gay and transgender families with inclusive non‐discriminatory care.
Behavioural modification of the cholinergic anti‐inflammatory response to C‐reactive protein in patients with hypertension
Authors: Nolan R. P. , Floras J. S. , Ahmed L. , Harvey P. J. , Hiscock N. , Hendrickx H. , Talbot D.
Published: January 2012

Abstract.  Nolan RP, Floras JS, Ahmed L, Harvey PJ, Hiscock N, Hendrickx H, Talbot D (University Health Network and University of Toronto; Women’s College Hospital, University of Toronto, Toronto, ON, Canada; and Unilever Discover, Colworth Science Park, Sharnbrook, UK). Behavioural modification of the cholinergic anti‐inflammatory response to C‐reactive protein in patients with hypertension. J Intern Med 2012; 272: 161–169.

Objectives.  A central hypothesis of the cholinergic anti‐inflammatory reflex model is that innate immune activity is inhibited by the efferent vagus. We evaluated whether changes in markers of tonic or reflex vagal heart rate modulation following behavioural intervention were associated inversely with changes in high‐sensitivity C‐reactive protein (hsCRP) or interleukin‐6 (IL‐6).

Design.  Subjects diagnosed with hypertension (n = 45, age 35–64 years, 53% women) were randomized to an 8‐week protocol of behavioural neurocardiac training (with heart rate variability biofeedback) or autogenic relaxation. Assessments before and after intervention included pro‐inflammatory factors (hsCRP, IL‐6), markers of vagal heart rate modulation [RR high‐frequency (HF) power within 0.15–0.40 Hz, baroreflex sensitivity and RR interval], conventional measures of lipoprotein cholesterol and 24‐h ambulatory systolic and diastolic blood pressure.

Results.  Changes in hsCRP and IL‐6 were not associated with changes in lipoprotein cholesterol or blood pressure. After adjusting for anti‐inflammatory drugs and confounding factors, changes in hsCRP related inversely to changes in HF power (β =−0.25±0.1, P = 0.02), baroreflex sensitivity (β = −0.33±0.7, P = 0.04) and RR interval (β = −0.001 ± 0.0004, P = 0.02). Statistically significant relationships were not observed for IL‐6.

Conclusions.  Changes in hsCRP were consistent with the inhibitory effect of increased vagal efferent activity on pro‐inflammatory factors predicted by the cholinergic anti‐inflammatory reflex model. Clinical trials for patients with cardiovascular dysfunction are warranted to assess whether behavioural interventions can contribute independently to the chronic regulation of inflammatory activity and to improved clinical outcomes.
Differential effect of transient global ischaemia on the levels of γ‐aminobutyric acid type A (GABAA) receptor subunit mRNAs in young and older rats
Authors: Montori S. , Dos_Anjos S. , Poole A. , Regueiro‐Purriños M. M. , Llorente I. L. , Darlison M. G. , Fernández‐López A. , Martínez‐Villayandre B.
Published: January 2012

S. Montori, S. Dos_Anjos, A. Poole, M. M. Regueiro‐Purriños, I. L. Llorente, M. G. Darlison, A. Fernández‐López and B. Martínez‐Villayandre (2012) Neuropathology and Applied Neurobiology38, 710–722

Differential effect of transient global ischaemia on the levels of γ‐aminobutyric acid type A (GABAA) receptor subunit mRNAs in young and older rats

Aims: This study has investigated how global brain ischaemia/reperfusion (I/R) modifies levels of mRNAs encoding γ‐aminobutyric acid type A (GABAA) receptor α1, β2 and γ2 subunits and glutamic acid decarboxylase 65 (GAD65) in an age‐ and structure‐dependent manner. Gene expression in response to treatment with the anti‐inflammatory agent meloxicam was also investigated. Methods: Global ischaemia was induced in 3‐ and 18‐month‐old male Sprague–Dawley rats. CA1, CA3, and dentate gyrus (DG) hippocampal areas, cerebral cortex (CC) and caudate putamen (C‐Pu) from sham‐operated and I/R‐injured animals were excised 48 h after the insult and prepared for quantitative polymerase chain reaction assays. Following I/R, meloxicam treatment was also carried out on young animals. Results: Data revealed significant decreases in the levels of all GABAA receptor subunit transcripts in the hippocampus of both young and older injured animals compared with sham‐operated ones. In contrast, there was either an increase or no change in GAD65 mRNA levels. GABAA receptor subunit transcript decreases were also observed in the CC and C‐Pu in young injured animals but not in the CC of the older injured ones; interestingly, significant increases were observed in the C‐Pu of older injured animals compared with controls. Meloxicam treatment following the insult resulted in a diminution of the previously described I/R response. Conclusions: The data indicate that I/R results in the modification of the levels of several gene transcripts involved in GABAergic signalling in both the pre‐ and postsynaptic components, of this neurotransmitter system, in an age‐ and structure‐dependent manner.
Comparison of extraction procedures for detection of Xanthomonas axonopodis pv. phaseoli in common bean seed
Authors: He Y. , Munkvold G. P.
Published: January 2012

Xanthomonas axonopodis pv. phaseoli (Xap) is an important seedborne pathogen of Phaseolus vulgaris. Accurate seed health testing methods are critical to protect seed quality and meet phytosanitary requirements. Currently employed selective media‐based methods include several variations in extraction procedures. In order to optimize pathogen extraction from seeds, the influence of different extraction steps on the sensitivity of Xap detection was assessed. Seeds were inoculated by vacuum infiltration with Xap to achieve inoculum levels from 101 to 105 CFU per seed; one contaminated seed was mixed into 1000‐seed subsamples of uncontaminated P. vulgaris seeds. Thirty subsamples of 1000 seeds were tested using each different extraction procedure. These included soaking whole seeds in sterilized saline phosphate buffer, either overnight at 4°C or for 3 h at room temperature, with or without vacuum extraction, and either with or without concentrating the seed extract by centrifuging. Seed extract dilutions were cultured on semiselective agar media MT and XCP1. The percentages of positive subsamples were compared to measure the effects of each extraction step on detection sensitivity. Vacuum extraction and centrifugation of seed extracts increased sensitivity; the highest sensitivity was obtained with the 3 h vacuum extraction followed by centrifugation. These results were confirmed with naturally infested seeds; Xap was detected in 48 of 70 samples using the 3 h vacuum extraction with centrifugation, whereas only 35 of 70 field samples tested positive using overnight soaking, a significant difference. The results suggest that these steps would be valuable modifications to the current method approved by the International Seed Testing Association (ISTA).
Influence of an injection system on the effect of activated soil steaming on Brassica juncea and the natural weed seedbank
Authors: PERUZZI A , RAFFAELLI M , FRASCONI C , FONTANELLI M , BÀRBERI P
Published: January 2012

Peruzzi A, Raffaelli M, Frasconi C, Fontanelli M & Bàrberi P (2012) Influence of an injection system on the effect of activated soil steaming on Brassica juncea and the natural weed seedbank. Weed Research 52, 140–152.

Soil steaming is a promising preventive method for weed control in high‐value cash crops, such as vegetables and flowers. A 2‐year experiment (2005–2006) was carried out to test the effect of five different systems of steam injection (surface or deep steam injection bar and three mixed systems at different steam distribution ratios between the surface and the deep bar: 1:2; 1:1 and 2:1) on an artificial infestation of Brassica juncea and on the natural weed seedbank. Treatments were carried out using steam alone or in association with two different exothermic compounds (KOH and CaO), distributed at two different rates (1000 and 4000 kg ha−1). The effects of treatments on the seedbank were analysed by the seedling emergence technique at three different soil depths (0–7, 7–14 and 14–21 cm). Non‐linear dose‐response curves were used to correlate weed emergence with soil temperature sum. Surface steaming was the most effective treatment (up to 100% weed control) at 0–7 cm depth. Deep steaming depleted the weed seedbank up to 95% in the deepest soil layer. The most homogeneous weed control effect across the soil profile was obtained with mixed systems, with differences depending on their distribution ratio. The new steam injection systems increased the versatility of this innovative soil steaming machine and confirmed its efficiency in reducing the weed seedbank, making it a promising replacement of methyl bromide fumigation before growing high‐value crops.
Reduced expression of CTR1 gene modulated by mitochondria causes enhanced ethylene response in cytoplasmic male‐sterile Brassica juncea
Authors: Liu Xunyan , Yang Xiaodong , Zhao Xuan , Yang Jinghua , Zhang Mingfang
Published: January 2012

We studied how mitochondria affect ethylene response via modulation of CTR1 expression in cytoplasmic male‐sterile (CMS) Brassica juncea. The expression of CTR1 gene was reduced in CMS compared with male‐fertile (MF) lines. We observed that hypocotyl and root lengths were shorter than in the MF line during germination in the dark. An enhanced ethylene response was observed in CMS plants as shown by the CMS and maintainer line phenotypes treated with 1‐aminocyclopropane‐1‐carboxylic acid. The phenotype in CMS plants could be recovered to the maintainer line when treated with Ag+. One ethylene response gene, plant defensin gene, was detected to be induced in CMS. The behavior of this phenotype could be mimicked by treating the maintainer line with antimycin A that disturbs mitochondrial function, which showed reduced length of hypocotyl and roots, and resulted in similar expression patterns of ethylene‐related genes as in CMS. The reduced length of hypocotyl and roots could be recovered to the maintainer line by treatment with gibberellic acid (GA3). In addition, the GA3 content was reduced in CMS plants and in the MF line treated with antimycin A. Ethylene treatment markedly affects GA3 content; however, GA3 did not significantly affect ethylene‐related gene expression in regards to regulation of hypocotyl and root length, which suggests that ethylene acts upstream via gibberellin to regulate hypocotyls and root development. Taken together, our results suggest a link between mitochondrial modulation of the ethylene and gibberellin pathway that regulates the development of hypocotyl and roots.