bluuebunny/crossref_metadata_2025 huggingface dataset:
PML Regulates Apoptosis at Endoplasmic Reticulum by Modulating Calcium Release
Authors: Giorgi Carlotta , Ito Keisuke , Lin Hui-Kuan , Santangelo Clara , Wieckowski Mariusz R. , Lebiedzinska Magdalena , Bononi Angela , Bonora Massimo , Duszynski Jerzy , Bernardi Rosa , Rizzuto Rosario , Tacchetti Carlo , Pinton Paolo , Pandolfi Pier Paolo
Published: October 2010

Promoting Apoptosis

During acute disease, the promyelocytic leukemia (PML) protein becomes fused to another protein as a result of a chromosomal translocation. This protein appears to have multiple and varied functions, including the ability to form distinctive complexes in the nucleus that suppress tumorigenesis and promote apoptotic cell death.

Giorgi
et al.

(p.
1247
, published online 28 October; see the Perspective by

Culjkovic-Kraljacic and Borden

) have proposed a mechanism by which PML influences the cellular signals that promote apoptosis. The protein was localized at sites of contact between the endoplasmic reticulum and mitochondria, where it associated with a calcium channel, a protein kinase, and a protein phosphatase, to regulate calcium mobilization into the mitochondrion, which then triggers the cell death program.
Calcium-Permeable AMPA Receptor Dynamics Mediate Fear Memory Erasure
Authors: Clem Roger L. , Huganir Richard L.
Published: October 2010

Wiping Out Memories

Inhibition of fear responses can be unexpectedly reversed even when a subject is perfectly safe. This can lead to inappropriate reactions to a fear-associated trigger, such as a bright light or loud noise. This type of reaction appears to underpin posttraumatic stress disorder, but there is little understanding of when training to inhibit fear may fail or succeed. Using a combination of electrophysiology and behavioral training in mice,
Clem and Huganir
(p.
1108
, published online 28 October) observed that fear conditioning increased synaptic transmission by calcium-permeable AMPA receptors into the part of the brain that controls emotional responses (the amygdala). This effect lasted for about a week, during which the fearful memories could be erased if the animals were trained to reduce conditioned fear responses. Postmortem brain slices showed that the fear-induced synaptic changes also reversed, except in transgenic mice with a mutant subunit of the AMPA receptor.
Politics and science in classifying the dangers of drugs
Authors: Room Robin , Lubman Dan I
Published: October 2010

There is a long history of psychoactive substances being regarded as dangerous and subsequently being banned or forbidden.1 Often the bans were introduced on substances new and unfamiliar to a society, which were viewed as more dangerous than substances which were well known and enculturated. With industrialisation and the globalisation brought by European empires, the growing availability of psychoactive substances was increasingly seen as a problem in the 1800s, setting off social and policy reactions – what we know as the temperance movement against alcohol,2 and initial UK legislation limiting the sale of ‘poisons’.3
Detection of clonality in follicular lymphoma using formalin-fixed, paraffin-embedded tissue samples and BIOMED-2 immunoglobulin primers
Authors: Berget Ellen , Helgeland Lars , Molven Anders , Vintermyr Olav Karsten
Published: October 2010

Aims

The BIOMED-2 multiplex PCR protocol is a commonly used procedure for assessing B cell clonality in lymphoma diagnostics. Follicular lymphoma poses a special challenge for PCR-based analyses because of high prevalence of somatic hypermutations in the rearranged immunoglobulin (IG) domains. This study aimed to evaluate the BIOMED-2 protocol performance in detection of B cell clonality in follicular lymphoma using formalin-fixed, paraffin-embedded (FFPE) tissue.

Methods

FFPE samples from 118 patients diagnosed with follicular lymphoma in the period 1998–2008 were used in the study. Clonality of IG heavy (IGH) and light chains (IGK,IGL) was assessed using a PCR procedure that was optimised for FFPE tissue.

Results

The highest clonal detection rates were 67.8% with theIGHVн-FR2-Jн assay and 66.1% with theIGKVκ-Jκ assay. Clonality was detected in 94.9% of all FFPE follicular lymphoma samples when all assays were combined. FFPE samples stored for 1–5 years did not perform significantly differently from those stored for 6–11 years. Interobserver agreement of clonality was tested for all analyses. The lowest score (Cohen'sκvalue = 0.56) was observed for theIGKVκ-Jκ clonality assay.

Conclusions

An improved PCR protocol for detection of clonality in FFPE samples using BIOMED-2 IG primers is presented. For best performance, a combination ofIGHandIGKanalyses is recommended.
Sensitive multiplex detection of KRAS codons 12 and 13 mutations in paraffin-embedded tissue specimens
Authors: Laosinchai-Wolf Walairat , Ye Fei , Tran Vu , Yang Zhe , White Roxanna , Bloom Kenneth , Choppa Paul , Labourier Emmanuel
Published: October 2010

Background

Colorectal cancer patients harbouring KRAS mutations in codon 12 or 13 do not benefit from current anti-epidermal growth factor receptor (EGFR) monoclonal antibody therapies. Efficient and robust methods are therefore required for routine clinical testing of KRAS mutation status.

Aims

To evaluate a novel multiplex assay for the rapid detection of common KRAS mutations in formalin-fixed paraffin-embedded (FFPE) tissues.

Methods

Genomic DNA was amplified by multiplex PCR using primers targeting the KRAS codon 12/13 region and an internal control gene. PCR products were hybridised on a liquid bead array containing target-specific probes and detected by particle flow cytometry.

Results

Analytical performance assessed with plasmid DNA and genomic DNA extracted from cell lines or model FFPE cell line dilutions showed specific detection of seven distinct KRAS mutations with a limit of detection equivalent to 1% tumour. The assay was evaluated at two independent sites with a total of 140 clinical specimens. At site 1, about 45% of the specimens from a set of 86 archived FFPE blocks with unknown KRAS mutation status were found positive for a KRAS mutation. At site 2, each of the seven mutations was detected in at least five independent specimens from a selected set of 54 residual genomic DNAs previously tested with an ARMS/Scorpion laboratory-developed test.

Conclusions

This novel single-well assay is a sensitive tool compatible with the clinical laboratory workflow for the rapid assessment of KRAS mutations in solid tumour specimens. Its performance and multiplex format warrant the development of broader panels including other relevant mutations in the EGFR pathway.
Comparative use of CO2laser, diode laser and monopolar electrocautery for resection of the soft palate in dogs with brachycephalic airway obstructive syndrome
Authors: Dunié‐Mérigot A. , Bouvy B. , Poncet C.
Published: October 2010

Clinical results, complications and the outcome of using either a carbon dioxide (CO2) laser, diode laser or electrocautery (ELEC) for resection of the soft palate with an extended palatoplasty technique in brachycephalic dogs with upper airway obstructive syndrome were compared. Dogs were randomly allocated into three groups (n=20 in each group): ELEC, diode and CO2groups. The palatoplasty was made at the rostral aspect of the tonsils. A respiratory clinical score, ranging from 0 (normal) to 4 (cyanosis), was attributed to each dog before surgery and at 0 hours, 24 hours, two weeks and six months after surgery. A favourable outcome was defined as a one point or greater decrease in score 24 hours after surgery. The proportion of dogs with a favourable outcome was significantly higher in the CO2(n=15) and ELEC groups (n=15) in comparison with the diode group (n=7) (OR=5.6, 95 per cent confidence interval 1.4 to 21.9). Surgical time was significantly shorter (P<0.001; mean [sd] 510 [178] seconds), and bleeding was less common (P<0.001; 30 per cent of cases) in the CO2group. Complications were most frequent with the diode group (two cases of death and two cases of tracheostomy). The final outcome for all groups (n=57) was considered excellent in 79 per cent of cases and was considered good in 21 per cent.
Mycobacterium bovis genotypes in Northern Ireland: herd‐level surveillance (2003 to 2008)
Authors: Skuce R. A. , Mallon T. R. , McCormick C. M. , McBride S. H. , Clarke G. , Thompson A. , Couzens C. , Gordon A. W. , McDowell S. W. J.
Published: October 2010

Surveillance genotyping (variable number tandem repeat profiling and spoligotyping) of Mycobacterium bovis isolates from culture‐confirmed bovine tuberculosis (TB)‐affected herds in Northern Ireland is presented for the years 2003 to 2008 inclusive. A total of 175 M bovis genotypes were identified in 8630 isolates from 6609 herds. On average, 73 genotypes were identified each year, with 29 genotypes present in all six years. Highly significant differences (P<0.0001) were observed between the relative frequency of some genotypes in the years 2003 to 2008. The spatial distribution of M bovis genotypes was not random (P<0.0001). Significant geographical localisation of M bovis genotypes was evident, suggesting that sources tended to be local. Despite regions being dominated by geographically localised genotypes, substantial and exploitable local diversity was still evident. Genotypes were also translocated significant distances from their normal geographical location.
More for less: dairy production in the 21st century
Authors: Huxley Jon , Green Martin
Published: October 2010

The concept of ‘more for less’ certainly applies in the dairy industry, but has it come at a cost in terms of animal health and welfare, ask Jon Huxley and Martin Green
A sequential method for selecting parameter values in the Barcelona basic model
Authors: Gallipoli Domenico , D’Onza Francesca , Wheeler Simon J.
Published: October 2010

The popularity of the Barcelona basic model (BBM) has grown steadily since its publication in 1990, due to its ability to capture key aspects of unsaturated soil behaviour. Nevertheless, the BBM is still rarely employed by practitioners, partly because of the absence of simple and objective methods for selecting parameter values from laboratory tests. One difficulty is that, in the BBM, individual aspects of the isotropic virgin behaviour are controlled by multiple parameters, while at the same time a single parameter controls more than one aspect of soil behaviour. This has led to iterative procedures where parameter values are adjusted in turn to match experiments, which requires significant experience and can lead to the selection of widely varying parameter values depending on the user. The proposed method streamlines parameter selection with a view to increasing the appeal of the BBM for practitioners. The method adopts a “sequential” procedure where the five parameters governing isotropic virgin behaviour are matched to degrees of freedom in the BBM, which are then fixed one at a time, in a specific order, without any assumption about other parameters. The simplicity and reduced subjectivity of the method in comparison with iterative procedures is demonstrated by selecting parameter values from laboratory tests.
Long-term settlement of soft subsoil clay under rectangular or semi-sinusoidal repeated loading of low amplitude
Authors: Hu Ya-Yuan
Published: October 2010

By applying Yin and Graham’s one-dimensional (1-D) equivalent-time rheological model, which can predict both primary and secondary settlement, the expressions for calculating the long-term subsoil settlement under rectangular or semi-sinusoidal repeated loading of low amplitude are obtained without considering the fluid–solid coupling. A property has been proven that ultimate settlement of clay is independent of stress increasing history under 1-D conditions. According to this property, the expressions are derived considering the solid–fluid coupling to approximately calculate the long-term total settlement and the long-term settlement of soft subsoil clay after construction of the road embankment. Because the expressions of the long-term settlement are analytical solutions in the present paper, they can not only avoid cumulative calculation errors in the numerical analysis, but also have more solid theoretical foundations than empirical equations.
Artificial neural network application for the prediction of ground surface movements induced by shield tunnelling
Authors: Boubou Rim , Emeriault Fabrice , Kastner Richard
Published: October 2010

This paper presents a methodology to correlate ground surface movements and tunnel boring machine (TBM) operation parameters. Two approaches are proposed and evaluated based on a case study of a shallow tunnel in a dense urban area. The first approach is based on a least square approximation and the second one uses an artificial neural network model. Data analysed were selected from the excavation of the subway line B tunnel in Toulouse, France, which was performed mainly by a shield TBM. Ground movements measured on the 4.7 km long contract 2 are reproduced with reasonable agreement by each of the two approaches. The amount of data (in particular for TBM operation parameters), the rather small amplitude of measured movements (a few millimetres), and the accuracy of these measurements (designed for routine construction management) make it necessary to create a pre-processing technique for the data, and a step-by-step improvement of approaches used. An elimination procedure is proposed to identify the most influential operation parameters and a sensitivity analysis shows their respective effect on ground movements.
Effects of infiltration and evaporation on geosynthetic capillary barrier performance
Authors: McCartney John S. , Zornberg Jorge G.
Published: October 2010

This study includes an experimental investigation of the transient movement of water in unsaturated soil layers underlain by a geocomposite drainage layer (GDL) during cycles of infiltration and evaporation. The distribution in volumetric water content with depth in a soil column having a height of 1350 mm underlain by a GDL was measured during transient infiltration. The capillary break effect was observed to affect the soil up to a height of 500 mm above the GDL, with an increase in volumetric water content up to 20% above that expected for the case of infiltration under a unit hydraulic gradient. Due to the long duration of this test (2000 h), a shorter 150 mm high soil column was also evaluated to investigate the soil–GDL hydraulic interaction during cycles of infiltration and evaporation. The capillary break was observed to have re-established itself after infiltration was stopped and the soil near the interface dried. The suction and volumetric water content measured in the soil at breakthrough were consistent after multiple cycles of wetting and drying. The conditions in the soil after each breakthrough event corresponded to the point on the drying-path water retention curve of the nonwoven geotextile where it transitioned from residual to saturated conditions.
Pseudo-static lateral earth pressures on broken-back retaining walls
Authors: Sadrekarimi Abouzar
Published: October 2010

Displacement of retaining walls during earthquakes causes damage to the structures founded on their backfill. The displacement of the wall can be reduced by decreasing the lateral earth pressure applied on its back. This can be achieved in a broken-back wall as the size of the failure wedge formed behind the wall is reduced; therefore, the calculation of lateral earth pressures is essential in assessing the safety of and designing broken-back retaining walls. In this study, a series of reduced-scale shaking table model experiments were performed on broken-back quay walls composed of concrete blocks with two different rear-face shapes. In comparison with a vertical-back wall, earth pressures increased at the upper forward (i.e., seaward) leaning rear-face segments of the wall, whereas they decreased at lower backward (i.e., landward) leaning elevations. Because of the wide application of the pseudo-static method of Mononobe–Okabe in engineering practice and design codes, lateral earth pressures have also been estimated using this approach. The comparison between the measured lateral earth pressures and those calculated using the Mononobe–Okabe method shows fairly good agreement in predicting the overall distribution of lateral active earth pressure during and after the shaking.
A WAVELET-BASED SPEAKER VERIFICATION ALGORITHM
Authors: LACERDA MICHEL ALVES , GUIDO RODRIGO CAPOBIANCO , DE SOUZA LEONARDO MENDES , ZULATO PAULO RICARDO FRANCHI , RIBEIRO JUSSARA , CHEN SHI-HUANG
Published: October 2010

This paper presents a study on wavelets and their characteristics for the specific purpose of serving as a feature extraction tool for speaker verification (SV), considering a Radial Basis Function (RBF) classifier, which is a particular type of Artificial Neural Network (ANN). Examining characteristics such as support-size, frequency and phase responses, amongst others, we show how Discrete Wavelet Transforms (DWTs), particularly the ones which derive from Finite Impulse Response (FIR) filters, can be used to extract important features from a speech signal which are useful for SV. Lastly, an SV algorithm based on the concepts presented is described.
Effect of adjuvant-induced systemic inflammation in rats on hepatic disposition kinetics of taurocholate
Authors: Roberts Michael S. , Liu Xin , Zou Yuhong , Siebert Gerhard A. , Chang Ping , Whitehouse Michael W. , Fletcher Linda , Crawford Darrell H. G.
Published: October 2010

It has been reported that the adjuvant-induced inflammation could affect drug metabolism in liver. Here we further investigated the effect of inflammation on drug transport in liver using taurocholate as a model drug. The hepatic disposition kinetics of [3H]taurocholate in perfused normal and adjuvant-treated rat livers were investigated by the multiple indicator dilution technique and data were analyzed by a previously reported hepatobiliary taurocholate transport model. Real-time RT-PCR was also performed to determine the mRNA expression of liver bile salt transporters in normal and diseased livers. The uptake and biliary excretion of taurocholate were impaired in the adjuvant-treated rats as shown by decreased influx rate constant kin(0.65 ± 0.09 vs. 2.12 ± 0.30) and elimination rate constant kbe(0.09 ± 0.02 vs. 0.17 ± 0.04) compared with control rat group, whereas the efflux rate constant koutwas greatly increased (0.07 ± 0.02 vs. 0.02 ± 0.01). The changes of mRNA expression of liver bile salt transporters were found in adjuvant-treated rats. Hepatic taurocholate extraction ratio in adjuvant-treated rats (0.86 ± 0.05, n = 6) was significantly reduced compared with 0.93 ± 0.05 ( n = 6) in normal rats. Hepatic extraction was well correlated with altered hepatic ATP content ( r2= 0.90). In conclusion, systemic inflammation greatly affects hepatic ATP content/production and associated transporter activities and causes an impairment of transporter-mediated solute trafficking and pharmacokinetics.
Apical Na+-d-glucose cotransporter 1 (SGLT1) activity and protein abundance are expressed along the jejunal crypt-villus axis in the neonatal pig
Authors: Yang Chengbo , Albin David M. , Wang Zirong , Stoll Barbara , Lackeyram Dale , Swanson Kendall C. , Yin Yulong , Tappenden Kelly A. , Mine Yoshinori , Yada Rickey Y. , Burrin Douglas G. , Fan Ming Z.
Published: October 2010

Gut apical Na+-glucose cotransporter 1 (SGLT1) activity is high at the birth and during suckling, thus contributing substantially to neonatal glucose homeostasis. We hypothesize that neonates possess high SGLT1 maximal activity by expressing apical SGLT1 protein along the intestinal crypt-villus axis via unique control mechanisms. Kinetics of SGLT1 activity in apical membrane vesicles, prepared from epithelial cells sequentially isolated along the jejunal crypt-villus axis from neonatal piglets by the distended intestinal sac method, were measured. High levels of maximal SGLT1 uptake activity were shown to exist along the jejunal crypt-villus axis in the piglets. Real-time RT-PCR analyses showed that SGLT1 mRNA abundance was lower ( P < 0.05) by 30–35% in crypt cells than in villus cells. There were no significant differences in SGLT1 protein abundances on the jejunal apical membrane among upper villus, middle villus, and crypt cells, consistent with the immunohistochemical staining pattern. Higher abundances ( P < 0.05) of total eukaryotic initiation factor 4E (eIF4E) protein and eIE4E-binding protein 1 γ-isoform in contrast to a lower ( P < 0.05) abundance of phosphorylated (Pi) eukaryotic elongation factor 2 (eEF2) protein and the eEF2-Pi to total eEF2 abundance ratio suggest higher global protein translational efficiency in the crypt cells than in the upper villus cells. In conclusion, neonates have high intestinal apical SGLT1 uptake activity by abundantly expressing SGLT1 protein in the epithelia and on the apical membrane along the entire crypt-villus axis in association with enhanced protein translational control mechanisms in the crypt cells.
Role of liver sinusoidal endothelial cells and stabilins in elimination of oxidized low-density lipoproteins
Authors: Li Ruomei , Oteiza Ana , Sørensen Karen Kristine , McCourt Peter , Olsen Randi , Smedsrød Bård , Svistounov Dmitri
Published: October 2010

Atherogenesis is associated with elevated levels of low-density lipoprotein (LDL) and its oxidized form (oxLDL) in the blood. The liver is an important scavenger organ for circulating oxLDLs. The present study aimed to examine endocytosis of mildly oxLDL (the major circulating form of oxLDLs) in liver sinusoidal endothelial cells (LSECs) and the involvement of the scavenger receptors stabilin-1 and stabilin-2 in this process. Freshly isolated LSECs, Kupffer cells (KCs), and stabilin-1- and stabilin-2-transfected human embryonic kidney cells were incubated with fluorescently labeled or radiolabeled oxLDLs [oxidized for 3 h (oxLDL3), 6 h, or 24 h (oxLDL24)] to measure endocytosis. The intracellular localization of oxLDLs and stabilins in LSECs was examined by immunofluorescence and immunogold electron microscopy. Whereas oxLDL24was endocytosed both by LSECs and KCs, oxLDL3(mildly oxLDL) was taken up by LSECs only. The LSEC uptake of oxLDLs was significantly inhibited by the scavenger receptor ligand formaldehyde-treated serum albumin. Uptake of all modified LDLs was high in stabilin-1-transfected cells, whereas stabilin-2-transfected cells preferentially took up oxLDL24, suggesting that stabilin-1 is a more important receptor for mildly oxLDLs than stabilin-2. Double immunogold labeling experiments in LSECs indicated interactions of stabilin-1 and stabilin-2 with oxLDL3on the cell surface, in coated pits, and endocytic vesicles. LSECs but not KCs endocytosed mildly oxLDL. Both stabilin-1 and stabilin-2 were involved in the LSEC endocytosis of oxLDLs, but experiments with stabilin-transfected cells pointed to stabilin-1 as the most important receptor for mildly oxLDL.
Physiological relevance of cell-specific distribution patterns of CFTR, NKCC1, NBCe1, and NHE3 along the crypt-villus axis in the intestine
Authors: Jakab Robert L. , Collaco Anne M. , Ameen Nadia A.
Published: October 2010

We examined the cell-specific subcellular expression patterns for sodium- and potassium-coupled chloride (NaK2Cl) cotransporter 1 (NKCC1), Na+bicarbonate cotransporter (NBCe1), cystic fibrosis transmembrane conductance regulator (CFTR), and Na+/H+exchanger 3 (NHE3) to understand the functional plasticity and synchronization of ion transport functions along the crypt-villus axis and its relevance to intestinal disease. In the unstimulated intestine, all small intestinal villus enterocytes coexpressed apical CFTR and NHE3, basolateral NBCe1, and mostly intracellular NKCC1. All (crypt and villus) goblet cells strongly expressed basolateral NKCC1 (at approximately three-fold higher levels than villus enterocytes), but no CFTR, NBCe1, or NHE3. Lower crypt cells coexpressed apical CFTR and basolateral NKCC1, but no NHE3 or NBCe1 (except NBCe1-expressing proximal colonic crypts). CFTR, NBCe1, and NKCC1 colocalized with markers of early and recycling endosomes, implicating endocytic recycling in cell-specific anion transport. Brunner's glands of the proximal duodenum coexpressed high levels of apical/subapical CFTR and basolateral NKCC1, but very low levels of NBCe1, consistent with secretion of Cl−-enriched fluid into the crypt. The cholinergic agonist carbachol rapidly (within 10 min) reduced cell volume along the entire crypt/villus axis and promoted NHE3 internalization into early endosomes. In contrast, carbachol induced membrane recruitment of NKCC1 and CFTR in all crypt and villus enterocytes, NKCC1 in all goblet cells, and NBCe1 in all villus enterocytes. These observations support regulated vesicle traffic in Cl−secretion by goblet cells and Cl−and HCO3−secretion by villus enterocytes during the transient phase of cholinergic stimulation. Overall, the carbachol-induced membrane trafficking profile of the four ion transporters supports functional plasticity of the small intestinal villus epithelium that enables it to conduct both absorptive and secretory functions.
Monosodium glutamate raises antral distension and plasma amino acid after a standard meal in humans
Authors: Boutry Claire , Matsumoto Hideki , Airinei Gheorghe , Benamouzig Robert , Tomé Daniel , Blachier François , Bos Cécile
Published: October 2010

The consumption of monosodium glutamate (MSG) is advocated to elicit physiological and metabolic effects, yet these effects have been poorly investigated directly in humans and in particular in the postprandial phase. Thirteen healthy adults were supplemented for 6 days with a nutritional dose of MSG (2 g) or sodium chloride (NaCl) as control, following a crossover design. On the 7th day, they underwent a complete postprandial examination for the 6 h following the ingestion of the same liquid standard meal (700 kcal, 20% of energy as [15N]protein, 50% as carbohydrate, and 30% as fat) supplemented with MSG or NaCl. Real-ultrasound measures of antral area indicated a significant increased distension for the 2 h following the meal supplemented with MSG vs. NaCl. This early postprandial phase was also associated with significantly increased levels of circulating leucine, isoleucine, valine, lysine, cysteine, alanine, tyrosine, and tryptophan after MSG compared with NaCl. No changes to the postprandial glucose, insulin, glucagon-like peptide (GLP)-1, and ghrelin were noted between MSG- and NaCl-supplemented meals. Subjective assessments of hunger and fullness were neither affected by MSG supplementation. Finally, the postprandial fate of dietary N was identical between dietary conditions. Our findings indicate that nutritional dose of MSG promoted greater postprandial elevations of several indispensable amino acids in plasma and induced gastric distension. Further work to elucidate the possible sparing effect of MSG on indispensable amino acid first-pass uptake in humans is warranted. This trial was registered at clinicaltrials.gov as NCT00862017.
Restoration of PPARγ reverses lipid accumulation in alveolar macrophages of GM-CSF knockout mice
Authors: Malur Anagha , Baker Anna D. , Mccoy Almedia J. , Wells Greg , Barna Barbara P. , Kavuru Mani S. , Malur Achut G. , Thomassen Mary Jane
Published: October 2010

Pulmonary alveolar proteinosis (PAP) is a lung disease characterized by a deficiency of functional granulocyte macrophage colony-stimulating factor (GM-CSF) resulting in surfactant accumulation and lipid-engorged alveolar macrophages. GM-CSF is a positive regulator of PPARγ that is constitutively expressed in healthy alveolar macrophages. We previously reported decreased PPARγ and ATP-binding cassette transporter G1 (ABCG1) levels in alveolar macrophages from PAP patients and GM-CSF knockout (KO) mice, suggesting PPARγ and ABCG1 involvement in surfactant catabolism. Because ABCG1 represents a PPARγ target, we hypothesized that PPARγ restoration would increase ABCG1 and reduce macrophage lipid accumulation. Upregulation of PPARγ was achieved using a lentivirus expression system in vivo. GM-CSF KO mice received intratracheal instillation of lentivirus (lenti)-PPARγ or control lenti-eGFP. Ten days postinstillation, 79% of harvested alveolar macrophages expressed eGFP, demonstrating transduction. Alveolar macrophages showed increased PPARγ and ABCG1 expression after lenti-PPARγ instillation, whereas PPARγ and ABCG1 levels remained unchanged in lenti-eGFP controls. Alveolar macrophages from lenti-PPARγ-treated mice also exhibited reduced intracellular phospholipids and increased cholesterol efflux to HDL, an ABCG1-mediated pathway. In vivo instillation of lenti-PPARγ results in: 1) upregulating ABCG1 and PPARγ expression of GM-CSF KO alveolar macrophages, 2) reducing intracellular lipid accumulation, and 3) increasing cholesterol efflux activity.
Muscle activity and time to task failure differ with load compliance and target force for elbow flexor muscles
Authors: Rudroff Thorsten , Justice Jamie N. , Holmes Matthew R. , Matthews Stephen D. , Enoka Roger M.
Published: October 2010

The primary purpose of this study was to determine the influence of load compliance on time to failure during sustained isometric contractions performed with the elbow flexor muscles at four submaximal target forces. Subjects pulled against a rigid restraint during the force task and maintained a constant elbow angle, while supporting an equivalent inertial load during the position task. Each task was sustained for as long as possible. Twenty-one healthy adults (23 ± 6 yr; 11 men) participated in the study. The maximal voluntary contraction (MVC) force was similar ( P = 0.95) before the subjects performed the force and position tasks at each of the four target forces: 20, 30, 45, and 60% of MVC force. The time to task failure was longer for the force tasks (576 ± 80 and 325 ± 70 s) than for the position tasks (299 ± 77 and 168 ± 35 s) at target forces of 20 and 30% ( P < 0.001), but was similar for the force tasks (178 ± 35 and 86 ± 14 s) and the position tasks (132 ± 29 and 87 ± 14 s) at target forces of 45 and 60% ( P > 0.19). The briefer times to failure for the position task at the lower forces were accompanied by greater rates of increase in elbow flexor muscle activity, mean arterial pressure, heart rate, and rating of perceived exertion. There was no difference in the estimates of external mechanical work at any target force. The dominant mechanisms limiting time to failure of sustained isometric contractions with the elbow flexor muscles appear to change at target forces between 30 and 45% MVC, with load compliance being a significant factor at lower forces only.
Differential effects of peroxynitrite on contractile protein properties in fast- and slow-twitch skeletal muscle fibers of rat
Authors: Dutka T. L. , Mollica J. P. , Lamb G. D.
Published: October 2010

Oxidative modification of contractile proteins is thought to be a key factor in muscle weakness observed in many pathophysiological conditions. In particular, peroxynitrite (ONOO−), a potent short-lived oxidant, is a likely candidate responsible for this contractile dysfunction. In this study ONOO− or 3-morpholinosydnonimine (Sin-1, a ONOO− donor) was applied to rat skinned muscle fibers to characterize the effects on contractile properties. Both ONOO− and Sin-1 exposure markedly reduced maximum force in slow-twitch fibers but had much less effect in fast-twitch fibers. The rate of isometric force development was also reduced without change in the number of active cross bridges. Sin-1 exposure caused a disproportionately large decrease in Ca2+ sensitivity, evidently due to coproduction of superoxide, as it was prevented by Tempol, a superoxide dismutase mimetic. The decline in maximum force with Sin-1 and ONOO− treatments could be partially reversed by DTT, provided it was applied before the fiber was activated. Reversal by DTT indicates that the decrease in maximum force was due at least in part to oxidation of cysteine residues. Ascorbate caused similar reversal, further suggesting that the cysteine residues had undergone S-nitrosylation. The reduction in Ca2+ sensitivity, however, was not reversed by either DTT or ascorbate. Western blot analysis showed cross-linking of myosin heavy chain (MHC) I, appearing as larger protein complexes after ONOO− exposure. The findings suggest that ONOO− initially decreases maximum force primarily by oxidation of cysteine residues on the myosin heads, and that the accompanying decrease in Ca2+ sensitivity is likely due to other, less reversible actions of hydroxyl or related radicals.
High responders to resistance exercise training demonstrate differential regulation of skeletal muscle microRNA expression
Authors: Davidsen Peter K. , Gallagher Iain J. , Hartman Joseph W. , Tarnopolsky Mark A. , Dela Flemming , Helge Jørn W. , Timmons James A. , Phillips Stuart M.
Published: October 2010

MicroRNAs (miRNA), small noncoding RNA molecules, may regulate protein synthesis, while resistance exercise training (RT) is an efficient strategy for stimulating muscle protein synthesis in vivo. However, RT increases muscle mass, with a very wide range of effectiveness in humans. We therefore determined the expression level of 21 abundant miRNAs to determine whether variation in these miRNAs was able to explain the variation in RT-induced gains in muscle mass. Vastus lateralis biopsies were obtained from the top and bottom ∼20% of responders from 56 young men who undertook a 5 day/wk RT program for 12 wk. Training-induced muscle mass gain was determined by dual-energy X-ray absorptiometry, and fiber size was evaluated by histochemistry. The expression level of each miRNA was quantified using TaqMan-based quantitative PCR, with the analysis carried out in a blinded manner. Gene ontology and target gene profiling were used to predict the potential biological implications. Of the 21 mature miRNAs examined, 17 were stable during RT in both groups. However, miR-378, miR-29a, miR-26a, and miR-451 were differentially expressed between low and high responders. miR-378, miR-29a, and miR-26a were downregulated in low responders and unchanged in high responders, while miR-451 was upregulated only in low responders. Interestingly, the training-induced change in miR-378 abundance was positively correlated with muscle mass gains in vivo. Gene ontology analysis of the target gene list of miR-378, miR-29a, miR-26a, and miR-451, from the weighted cumulative context ranking methodology, indicated that miRNA changes in the low responders may be compensatory, reflecting a failure to “activate” growth and remodeling genes. We report, for the first time, that RT-induced hypertrophy in human skeletal muscle is associated with selected changes in miRNA abundance. Our analysis indicates that miRNAs may play a role in the phenotypic change and pronounced intergroup variation in the RT response.
Effect of anti-inflammatory medication on the running-induced rise in patella tendon collagen synthesis in humans
Authors: Christensen Britt , Dandanell Sune , Kjaer Michael , Langberg Henning
Published: October 2010

NSAIDs are widely used in the treatment of inflammatory diseases as well as of tendon diseases associated with pain in sports and labor. However, the effect of NSAID intake, and thus blockade of PGE2production, on the tendon tissue adaptation is unknown. The purpose of the present study was to elucidate the possible effects of NSAID intake on healthy tendon collagen turnover in relation to a strenuous bout of endurance exercise. Fifteen healthy young men were randomly assigned into two experimental groups, with one group receiving indomethacin (oral 2 × 100 mg Confortid daily for 7 days; NSAID; n = 7) and a placebo group ( n = 8). Both groups were exposed to a prolonged bout of running (36 km). The collagen synthesis NH2-terminal propeptide of type I (PINP) and PGE2concentrations were measured before and 72 h following the run in the patella tendon by microdialysis. The peritendinous concentrations of PINP increased significantly in the placebo group as a result of the run, as shown previously. PGE2levels were significantly decreased 72 h after the run compared with basal levels in the subjects treated with NSAID and unchanged in the placebo group. The NSAID intake abolished the adaptive increase in collagen synthesis in the patella tendon found in the placebo group in response to the prolonged exercise ( P < 0.05). The present study demonstrates that intake of NSAID decreased interstitial PGE2and abolished the exercise-induced adaptive increase in collagen synthesis in human tendons.
Nutritional modulation of training-induced skeletal muscle adaptations
Authors: Hawley John A. , Burke Louise M. , Phillips Stuart M. , Spriet Lawrence L.
Published: October 2010

Skeletal muscle displays remarkable plasticity, enabling substantial adaptive modifications in its metabolic potential and functional characteristics in response to external stimuli such as mechanical loading and nutrient availability. Contraction-induced adaptations are determined largely by the mode of exercise and the volume, intensity, and frequency of the training stimulus. However, evidence is accumulating that nutrient availability serves as a potent modulator of many acute responses and chronic adaptations to both endurance and resistance exercise. Changes in macronutrient intake rapidly alter the concentration of blood-borne substrates and hormones, causing marked perturbations in the storage profile of skeletal muscle and other insulin-sensitive tissues. In turn, muscle energy status exerts profound effects on resting fuel metabolism and patterns of fuel utilization during exercise as well as acute regulatory processes underlying gene expression and cell signaling. As such, these nutrient-exercise interactions have the potential to activate or inhibit many biochemical pathways with putative roles in training adaptation. This review provides a contemporary perspective of our understanding of the molecular and cellular events that take place in skeletal muscle in response to both endurance and resistance exercise commenced after acute and/or chronic alterations in nutrient availability (carbohydrate, fat, protein, and several antioxidants). Emphasis is on the results of human studies and how nutrient provision (or lack thereof) interacts with specific contractile stimulus to modulate many of the acute responses to exercise, thereby potentially promoting or inhibiting subsequent training adaptation.
Highly Purified Eicosapentaenoic Acid as Free Fatty Acids Strongly Suppresses Polyps in ApcMin/+ Mice
Authors: Fini Lucia , Piazzi Giulia , Ceccarelli Claudio , Daoud Yahya , Belluzzi Andrea , Munarini Alessandra , Graziani Giulia , Fogliano Vincenzo , Selgrad Michael , Garcia Melissa , Gasbarrini Antonio , Genta Robert M. , Boland C. Richard , Ricciardiello Luigi
Published: October 2010

Purpose: Although cyclooxygenase (COX)-2 inhibitors could represent the most effective chemopreventive tool against colorectal cancer (CRC), their use in clinical practice is hampered by cardiovascular side effects. Consumption of ω-3-polyunsaturated fatty acids (ω-3-PUFAs) is associated with a reduced risk of CRC. Therefore, in this study, we assessed the efficacy of a novel 99% pure preparation of ω-3-PUFA eicosapentaenoic acid as free fatty acids (EPA-FFA) on polyps in ApcMin/+ mice.

Experimental design: ApcMin/+ and corresponding wild-type mice were fed control diet (Ctrl) or diets containing either EPA-FFA 2.5% or 5%, for 12 weeks while monitoring food intake and body weight.

Results: We found that both EPA-FFA diets protected from the cachexia observed among ApcMin/+ animals fed Ctrl diet (P < 0.0054), without toxic effect, in conjunction with a significant decrease in lipid peroxidation in the treated arms. Moreover, both EPA-FFA diets dramatically suppressed polyp number (by 71.5% and 78.6%, respectively; P < 0.0001) and load (by 82.5% and 93.4%, respectively; P < 0.0001) in both small intestine and colon. In addition, polyps less than 1 mm in size were predominantly found in the EPA-FFA 5% arm whereas those 1 to 3 mm in size were more frequent in the Ctrl arm (P < 0.0001). Interestingly, in the EPA-FFA groups, mucosal arachidonic acid was replaced by EPA (P < 0.0001), leading to a significant reduction in COX-2 expression and β-catenin nuclear translocation. Moreover, in the EPA-FFA arms, we found a significant decrease in proliferation throughout the intestine together with an increase in apoptosis.

Conclusions: Our data make 99% pure EPA-FFA an excellent candidate for CRC chemoprevention. Clin Cancer Res; 16(23); 5703–11. ©2010 AACR.
Bevacizumab plus Fotemustine as First-line Treatment in Metastatic Melanoma Patients: Clinical Activity and Modulation of Angiogenesis and Lymphangiogenesis Factors
Authors: Del Vecchio Michele , Mortarini Roberta , Canova Stefania , Di Guardo Lorenza , Pimpinelli Nicola , Sertoli Mario R. , Bedognetti Davide , Queirolo Paola , Morosini Paola , Perrone Tania , Bajetta Emilio , Anichini Andrea
Published: October 2010

Purpose: To assess the clinical and biological activity of the association of bevacizumab and fotemustine as first-line treatment in advanced melanoma patients.

Experimental Design: Previously untreated, metastatic melanoma patients (n = 20) received bevacizumab (at 15 mg/kg every 3 weeks) and fotemustine (100 mg/m2 by intravenous administration on days 1, 8, and 15, repeated after 4 weeks) in a multicenter, single-arm, open-label, phase II study. Primary endpoint was the best overall response rate; other endpoints were toxicity, time to progression (TTP), and overall survival (OS). Serum cytokines, angiogenesis, and lymphangiogenesis factors were monitored by multiplex arrays and by in vitro angiogenesis assays. Effects of fotemustine on melanoma cells, in vitro, on vascular endothelial growth factor (VEGF)-C release and apoptosis were assessed by ELISA and flow cytometry, respectively.

Results: One complete response, 2 partial responses (PR), and 10 patients with stable disease were observed. TTP and OS were 8.3 and 20.5 months, respectively. Fourteen patients experienced adverse events of toxicity grade 3–4. Serum VEGF-A levels in evaluated patients (n = 15) and overall serum proangiogenic activity were significantly inhibited. A significant reduction in VEGF-C levels was found in several post-versus pretherapy serum samples. In vitro, fotemustine inhibited VEGF-C release by melanoma cells without inducing significant cell death. Serum levels of interleukin (IL)-10 and IL-12p70 showed the highest levels in sera of PR patients, compared with patients with stable or progressive disease whereas IL-23 showed the opposite pattern.

Conclusions: The combination of bevacizumab plus fotemustine has clinical activity in advanced melanoma and promotes systemic modulation of angiogenesis and lymphangiogenesis factors. Clin Cancer Res; 16(23); 5862–72. ©2010 AACR.
The Effect of Processing Speed on Cognitive Functioning in Patients with Familial Bipolar I Disorder and Their Unaffected Relatives
Authors: Antila Mervi , Kieseppä Tuula , Partonen Timo , Lönnqvist Jouko , Tuulio-Henriksson Annamari
Published: October 2010

Background: Despite increasing evidence of cognitive dysfunctions in bipolar I disorder, there is no specific neuropsychological profile of the disorder. Sampling and Method: The aim of the present study was to investigate the effect of processing speed on other cognitive functions in a population-based sample of 32 familial bipolar I disorder patients, their 40 unaffected first-degree relatives and 55 controls. A neuropsychological test battery was administered to all participants, and the effect of processing speed on other cognitive functions was analyzed with the digit symbol subtest of the Wechsler Adult Intelligence Scale-Revised both in within- and between-group comparisons. Results: After adjusting for the effect of processing speed, only small differences were detected in short-delay cued recall and in long-delay memory between patients and controls, as well as between patients and relatives. Relatives scored better than controls only in verbal ability. Processing speed had a significant effect on nearly all scores, differing by group when patients, relatives and controls were examined separately, the effect being most extensive in patients. Conclusions: These results support the view that impaired processing speed in particular contributes to a range of cognitive dysfunctions in bipolar disorder. However, it may not be specific to bipolar I disorder and can possibly be considered a shared endophenotype with other mental disorders.
Bioluminescence as Gold Standard for Validation of Optical Imaging Modalities in Peritoneal Carcinomatosis Animal Models
Authors: Harlaar N.J. , Hesselink J.W. , de Jong J.S. , van Dam G.M.
Published: October 2010

Background: The outcome of cytoreductive surgery in patients with peritoneal carcinomatosis is influenced by incomplete resection as a result of inadequate detection of a tumor, i.e. residual disease. The future perspective of complete resection, made possible by application of intraoperative near-infrared fluorescence imaging (NIRF), led to the development and validation of a bioluminescent colorectal peritoneal carcinomatosis xenograft rat model to act as the gold standard for the evaluation of new optical imaging modalities. Methods: Twenty nude rats were inoculated intraperitoneally with 2 × 106 luciferase-labeled human colorectal tumor cells (HT-29-luc-D6). The peritoneal carcinomatosis index (PCI) was estimated using visual observation (PCI-VO) and VO combined with bioluminescence imaging (PCI-BLI). Subsequently, the BL images were presented, and residual tumor tissue was localized by PCI-BLI scoring and compared with the PCI-VO. Results: BLI revealed additional tumor tissue, confirmed by HE staining, compared to VO alone in 7 out of 8 rats (p < 0.02). Conclusion: The developed model turned out to be suitable. The use of BLI for tumor detection was more sensitive compared to VO alone. In this model, BLI significantly detected residual disease, and therefore, BLI can be denominated as the gold standard for the evaluation of optical imaging modalities like NIRF.
Cytogenetic and Genetic Studies in a Hypospadic Horse (Equus caballus, 2n = 64)
Authors: De Lorenzi L. , Genualdo V. , Iannuzzi A. , Di Meo G.P. , Perucatti A. , Mancuso R. , Russo M. , Di Berardino D. , Parma P. , Iannuzzi L.
Published: October 2010

A 4-year-old male horse of Friesian breed with normal body conformation, development and libido, and showing an evident ventral penis deviation with hypospadias, underwent both cytogenetic and genetic investigation. Although the karyotype showed normal male arrangement (2n = 64,XY), one telomere of horse (ECA) chromosome 1 was shorter than both the other one and those of a normal horse (control), as revealed by CBA- and RBA-banding, and by Ag-NOR and FISH-mapping techniques using telomere PNA probes. Genetic investigation of the SRY and MAMLD1 coding sequences revealed a normal SRY sequence and a mutation in the MAMLD1 gene sequence: a homozygous change (C>A) was found, leading to the synthesis of an isoleucine, instead of a leucine. Although it is difficult to find a strict correlation between hypospadias and the genetic defects revealed by this investigation, this study is the first to be performed in a hypospadic horse using both cytogenetic and genetic investigation.
Mutations of the SRY-Responsive Enhancer of SOX9 Are Uncommon in XY Gonadal Dysgenesis
Authors: Georg I. , Bagheri-Fam S. , Knower K.C. , Wieacker P. , Scherer G. , Harley V.R.
Published: October 2010

During mouse sex determination, SRY upregulates the core testis-specific enhancer of Sox9, TESCO. Mutations in human SRY are found in one third of cases with XY pure gonadal dysgenesis (XY GD; Swyer syndrome), while two thirds remain unexplained. Heterozygous SOX9 mutations can cause XY GD in association with the skeletal malformation syndrome campomelic dysplasia. We hypothesized that human TESCO mutations could cause isolated XY GD. Sixty-six XY GD cases with an intact SRY were analyzed for TESCO point mutations or deletions. No mutations were identified. We conclude that TESCO mutations are not a common cause of XY GD.