bluuebunny/crossref_metadata_2025 huggingface dataset:
Image guided dose escalated prostate radiotherapy: still room to improve
Authors: Martin Jarad M , Bayley Andrew , Bristow Robert , Chung Peter , Gospodarowicz Mary , Menard Cynthia , Milosevic Michael , Rosewall Tara , Warde Padraig R , Catton Charles N
Published: November 2009

Background

Prostate radiotherapy (RT) dose escalation has been reported to result in improved biochemical control at the cost of greater late toxicity. We report on the application of 79.8 Gy in 42 fractions of prostate image guided RT (IGRT). The primary objective was to assess 5-year biochemical control and potential prognostic factors by the Phoenix definition. Secondary endpoints included acute and late toxicity by the Radiotherapy Oncology Group (RTOG) scoring scales.

Methods

From October/2001 and June/2003, 259 men were treated with at least 2-years follow-up. 59 patients had low, 163 intermediate and 37 high risk disease. 43 had adjuvant hormonal therapy (HT), mostly for high- or multiple risk factor intermediate-risk disease (n = 25). They received either 3-dimensional conformal RT (3DCRT, n = 226) or intensity modulated RT (IMRT) including daily on-line IGRT with intraprostatic fiducial markers.

Results

Median follow-up was 67.8 months (range 24.4-84.7). There was no severe (grade 3-4) acute toxicity, and grade 2 acute gastrointestinal (GI) toxicity was unusual (10.1%). The 5-year incidence of grade 2-3 late GI and genitourinary (GU) toxicity was 13.7% and 12.1%, with corresponding grade 3 figures of 3.5% and 2.0% respectively. HT had an association with an increased risk of grade 2-3 late GI toxicity (11% v 21%, p = 0.018). Using the Phoenix definition for biochemical failure, the 5 year-bNED is 88.4%, 76.5% and 77.9% for low, intermediate and high risk patients respectively. On univariate analysis, T-category and Gleason grade correlated with Phoenix bNED (p = 0.006 and 0.039 respectively). Hormonal therapy was not a significant prognostic factor on uni- or multi-variate analysis. Men with positive prostate biopsies following RT had a lower chance of bNED at 5 years (34.4% v 64.3%; p = 0.147).

Conclusion

IGRT to 79.8 Gy results in favourable rates of late toxicity compared with published non-IGRT treated cohorts. Future avenues of investigation for toxicity reduction include IMRT, margin reduction, and dose modulation targeted to sites of disease burden. Further work is required to maximize efficacy beyond that achieved through radiation dose escalation alone.
Recurrent microdeletion at 17q12 as a cause of Mayer-Rokitansky-Kuster-Hauser (MRKH) syndrome: two case reports
Authors: Bernardini Laura , Gimelli Stefania , Gervasini Cristina , Carella Massimo , Baban Anwar , Frontino Giada , Barbano Giancarlo , Divizia Maria Teresa , Fedele Luigi , Novelli Antonio , Béna Frédérique , Lalatta Faustina , Miozzo Monica , Dallapiccola Bruno
Published: November 2009

Background

Mayer-Rokitansky-Kuster-Hauser syndrome (MRKH) consists of congenital aplasia of the uterus and the upper part of vagina due to anomalous development of Müllerian ducts, either isolated or associated with other congenital malformations, including renal, skeletal, hearing and heart defects. This disorder has an incidence of approximately 1 in 4500 newborn girls and the aetiology is poorly understood.

Methods and Results

we report on two patients affected by MRKH syndrome in which array-CGH analysis disclosed an identical deletion spanning 1.5 Mb of genomic DNA at chromosome 17q12. One patient was affected by complete absence of uterus and vagina, with bilaterally normal ovaries, while the other displayed agenesis of the upper part of vagina, right unicornuate uterus, non cavitating rudimentary left horn and bilaterally multicystic kidneys. The deletion encompassed two candidate genes, TCF2 and LHX1. Mutational screening of these genes in a selected group of 20 MRKH females without 17q12 deletion was negative.

Conclusion

Deletion 17q12 is a rare albeit recurrent anomaly mediated by segmental duplications, previously reported in subjects with developmental kidney abnormalities and diabetes. The present two patients expand the clinical spectrum associated with this imbalance and suggest that this region is a candidate locus for a subset of MRKH syndrome individuals, with or without renal defects.
Unusual orbital lymphoma undetectable by magnetic resonance imaging: a case report
Authors: Tatsugawa Maria , Noma Hidetaka , Mimura Tatsuya , Funatsu Hideharu
Published: November 2009

Introduction

We report the case of a patient with orbital malignant lymphoma that was not detected by imaging studies when she presented with impaired vision, which lead to her eventual loss of sight.

Case presentation

A 71-year-old Japanese woman complained of deteriorating vision in her left eye. On examination, papilledema was detected, but magnetic resonance imaging only showed slight thickening and enhancement of the left optic nerve. A diagnosis of idiopathic optic neuritis was made and corticosteroid pulse therapy was administered. During the next four months, the patient received a total of four courses of corticosteroid pulse therapy, but she still suffered from bilateral loss of vision. A second magnetic resonance imaging procedure revealed tumors in both orbits and a biopsy showed diffuse large B-cell malignant lymphoma.

Conclusion

The possibility of malignant lymphoma should be considered in patients with recurrent optic neuropathy despite administration of corticosteroid pulse therapy, even when there are no abnormalities on cerebrospinal fluid examination or magnetic resonance imaging.
Laboratory-scale method for enzymatic saccharification of lignocellulosic biomass at high-solids loadings
Authors: Roche Christine M , Dibble Clare J , Stickel Jonathan J
Published: November 2009

Background

Screening new lignocellulosic biomass pretreatments and advanced enzyme systems at process relevant conditions is a key factor in the development of economically viable lignocellulosic ethanol. Shake flasks, the reaction vessel commonly used for screening enzymatic saccharifications of cellulosic biomass, do not provide adequate mixing at high-solids concentrations when shaking is not supplemented with hand mixing.

Results

We identified roller bottle reactors (RBRs) as laboratory-scale reaction vessels that can provide adequate mixing for enzymatic saccharifications at high-solids biomass loadings without any additional hand mixing. Using the RBRs, we developed a method for screening both pretreated biomass and enzyme systems at process-relevant conditions. RBRs were shown to be scalable between 125 mL and 2 L. Results from enzymatic saccharifications of five biomass pretreatments of different severities and two enzyme preparations suggest that this system will work well for a variety of biomass substrates and enzyme systems. A study of intermittent mixing regimes suggests that mass transfer limitations of enzymatic saccharifications at high-solids loadings are significant but can be mitigated with a relatively low amount of mixing input.

Conclusion

Effective initial mixing to promote good enzyme distribution and continued, but not necessarily continuous, mixing is necessary in order to facilitate high biomass conversion rates. The simplicity and robustness of the bench-scale RBR system, combined with its ability to accommodate numerous reaction vessels, will be useful in screening new biomass pretreatments and advanced enzyme systems at high-solids loadings.
microRNAs and genetic diseases
Authors: Meola Nicola , Gennarino Vincenzo Alessandro , Banfi Sandro
Published: November 2009

microRNAs (miRNAs) are a class of small RNAs (19-25 nucleotides in length) processed from double-stranded hairpin precursors. They negatively regulate gene expression in animals, by binding, with imperfect base pairing, to target sites in messenger RNAs (usually in 3' untranslated regions) thereby either reducing translational efficiency or determining transcript degradation. Considering that each miRNA can regulate, on average, the expression of approximately several hundred target genes, the miRNA apparatus can participate in the control of the gene expression of a large quota of mammalian transcriptomes and proteomes. As a consequence, miRNAs are expected to regulate various developmental and physiological processes, such as the development and function of many tissue and organs. Due to the strong impact of miRNAs on the biological processes, it is expected that mutations affecting miRNA function have a pathogenic role in human genetic diseases, similar to protein-coding genes. In this review, we provide an overview of the evidence available to date which support the pathogenic role of miRNAs in human genetic diseases. We will first describe the main types of mutation mechanisms affecting miRNA function that can result in human genetic disorders, namely: (1) mutations affecting miRNA sequences; (2) mutations in the recognition sites for miRNAs harboured in target mRNAs; and (3) mutations in genes that participate in the general processes of miRNA processing and function. Finally, we will also describe the results of recent studies, mostly based on animal models, indicating the phenotypic consequences of miRNA alterations on the function of several tissues and organs. These studies suggest that the spectrum of genetic diseases possibly caused by mutations in miRNAs is wide and is only starting to be unravelled.
Synergistic chondroprotective effects of curcumin and resveratrol in human articular chondrocytes: inhibition of IL-1β-induced NF-κB-mediated inflammation and apoptosis
Authors: Csaki Constanze , Mobasheri Ali , Shakibaei Mehdi
Published: November 2009

Introduction

Currently available treatments for osteoarthritis (OA) are restricted to nonsteroidal anti-inflammatory drugs, which exhibit numerous side effects and are only temporarily effective. Thus novel, safe and more efficacious anti-inflammatory agents are needed for OA. Naturally occurring polyphenolic compounds, such as curcumin and resveratrol, are potent agents for modulating inflammation. Both compounds mediate their effects by targeting the NF-κB signalling pathway.

Methods

We have recently demonstrated that in chondrocytes resveratrol modulates the NF-κB pathway by inhibiting the proteasome, while curcumin modulates the activation of NF-κB by inhibiting upstream kinases (Akt). However, the combinational effects of these compounds in chondrocytes has not been studied and/or compared with their individual effects. The aim of this study was to investigate the potential synergistic effects of curcumin and resveratrol on IL-1β-stimulated human chondrocytes in vitro using immunoblotting and electron microscopy.

Results

Treatment with curcumin and resveratrol suppressed NF-κB-regulated gene products involved in inflammation (cyclooxygenase-2, matrix metalloproteinase (MMP)-3, MMP-9, vascular endothelial growth factor), inhibited apoptosis (Bcl-2, Bcl-xL, and TNF-α receptor-associated factor 1) and prevented activation of caspase-3. IL-1β-induced NF-κB activation was suppressed directly by cocktails of curcumin and resveratrol through inhibition of Iκκ and proteasome activation, inhibition of IκBα phosphorylation and degradation, and inhibition of nuclear translocation of NF-κB. The modulatory effects of curcumin and resveratrol on IL-1β-induced expression of cartilage specific matrix and proinflammatory enzymes were mediated in part by the cartilage-specific transcription factor Sox-9.

Conclusions

We propose that combining these natural compounds may be a useful strategy in OA therapy as compared with separate treatment with each individual compound.
Effect of small interference RNA (siRNA) for ADAMTS5on intervertebral disc degeneration in the rabbit anular needle-puncture model
Authors: Seki Shoji , Asanuma-Abe Yumiko , Masuda Koichi , Kawaguchi Yoshiharu , Asanuma Kunihiro , Muehleman Carol , Iwai Akiko , Kimura Tomoatsu
Published: November 2009

Introduction

The etiology of degenerative disc disease is unknown. Several investigators have reported the presence of proteolytic enzymes, such as the matrix metalloproteinase (MMP) and ADAMTS (a disintegrin and metalloprotease with thrombospondin-like repeats) families, in degenerated human discs. Glasson and colleagues recently reported that a significant reduction occurs in the severity of cartilage destruction in ADAMTS5 knockout mice compared with wild-type mice. The purpose of this study was to evaluate the suppressive effects of injections of ADAMTS5 small interference RNA (siRNA) oligonucleotide on intervertebral disc degeneration in the rabbit anular needle-puncture model.

Methods

Rabbit nucleus pulposus (NP) cells were transfected with siRNA oligonucleotides specific for ADAMTS5 or the control. The suppression of the ADAMTS5 gene by siRNA transfection was assessed by using real-time polymerase chain reaction (PCR), both in monolayer and alginate bead cultures with or without interleukin-1β (IL-1β) stimulation. The effect of siRNA was determined in vivo by using the rabbit anular needle-puncture model (control group: n = 8; ADAMTS5 group: n = 8). One week after the initial anular puncture, the animals received an injection of the control or anti-ADAMTS5 oligonucleotide (100 μg each at the L2/3 and L4/5 level; 16 discs/group). Disc height, magnetic resonance imaging (MRI) (Thompson classification and signal intensity), and safranin-O staining (histologic grade) were assessed.

Results

IL-1β treatment significantly increased the ADAMTS5 mRNA level in NP cells (P < 0.01). ADAMTS5 gene suppression was 70% compared with the control oligonucleotide in both monolayer and alginate bead culture with or without stimulation with IL-1β. The injection of anti-ADAMTS5 oligonucleotide in vivo resulted in improved MRI scores with increased signal intensity and improved histologic grade scores with statistical significance (P < 0.05). No significant change in disc height was observed.

Conclusions

A single injection of ADAMTS5 siRNA induced the suppression of degradation in NP tissues, as shown by significantly improved MRI and histologic grades. The mechanism of response to siRNA may be worthy of exploration for possible therapeutic purposes.
Multidisciplinary care for tracheostomy patients: a systematic review
Authors: Garrubba Marie , Turner Tari , Grieveson Clare
Published: November 2009

Introduction

Appropriate care for patients with tracheostomies in hospital settings is an important issue. Each year more than 7000 patients receive tracheostomies in Australia and New Zealand alone. Many of these tracheostomy patients commence their care in the intensive care unit (ICU) and once stabilised are then transferred to a general ward. Insufficient skills and experience of staff caring for tracheostomy patients may lead to sub-optimal care and increased morbidity. The purpose of this review was to identify whether multidisciplinary tracheostomy outreach teams enable the reduction in time to decannulation and length of stay in acute and sub-acute settings, improve quality of care or decrease adverse events for patients with a tracheostomy.

Methods

We included all relevant trials published in English. We searched Medline, CINAHL, All EBM and EMBASE in June 2009. Studies were selected and appraised by two reviewers in consultation with colleagues, using inclusion, exclusion and appraisal criteria established a priori.

Results

Three studies were identified which met the study selection criteria. All were cohort studies with historical controls. All studies included adult patients with tracheostomies. One study was conducted in the UK and the other two in Australia. Risk of bias was moderate to high in all studies. All papers concluded that the introduction of multidisciplinary care reduces the average time to decannulation for tracheostomy patients discharged from the ICU. Two papers also reported that multidisciplinary care reduced the overall length of stay in hospital as well as the length of stay following ICU discharge.

Conclusions

In the papers we appraised, patients with a tracheostomy tube in situ discharged from an ICU to a general ward who received care from a dedicated multidisciplinary team as compared with standard care showed reductions in time to decannulation, length of stay and adverse events. Impacts on quality of care were not reported.

These results should be interpreted with caution due to the methodological weaknesses in the historical control studies.
The Response of Plasma Catecholamines in Rats Simultaneously Exposed to Immobilization and Painful Stimuli
Authors: Mravec Boris , Tillinger Andrej , Bodnar Ibolya , Nagy György M. , Palkovits Miklos , Kvetňanský Richard
Published: November 2009

Immobilization represents a strong stressor inducing a profound increase in plasma epinephrine and norepinephrine levels. We have previously demonstrated that a subcutaneous injection of formalin (0.2 mL of 4% solution/100 g bw) attenuated the immobilization‐induced elevation of plasma epinephrine levels in rats. In the present study, we investigated whether other painful and stressful stimuli, such as capsaicin, hydrochloric acid, mechanical pressure, heat, and cold, might also attenuate the increase of plasma epinephrine in rats exposed to acute immobilization stress. With the exception of formalin, all of the painful stimuli applied failed to attenuate the increase of plasma epinephrine levels in immobilized animals. Our data suggest that the attenuation of an immobilization‐induced increase in plasma epinephrine levels is specific for subcutaneous formalin administration.
Heart Adrenoceptor Gene Expression and Binding Sites in the Human Failing Heart
Authors: Fischer Viliam , Gabauer Ivan , Tillinger Andrej , Novakova Martina , Pechan Ivan , Krizanova Olga , Kvetňanský Richard , Myslivecek Jaromir
Published: November 2009

Adrenergic regulation of the heart function is well documented by many studies. Catecholamines act through α1‐, β1‐, β2‐, and β3‐adrenoceptors (ARs) in the heart. There are many findings about the changes of β1‐ and β2‐AR in heart failure (HF). On the other hand, the role of other AR subtypes is not clear yet. We focused on determining how HF could affect gene expression and specific ligand binding to α1A‐, α1B‐, α1D‐, β1‐, β2‐, and β3‐AR. Hearts from 11 patients with HF subjected to transplantation were investigated. As a control, corresponding parts from hearts not suitable for transplantation were used. We have found significantly higher mRNA levels of α1A‐, α1B‐,β1‐, and β2‐AR in the left ventricle of failing hearts compared to the levels in controls. β3‐AR mRNA levels in the left ventricle of failing hearts were not changed. No changes in mRNA levels of all receptors studied in other cardiac areas were found. On the other hand, binding studies showed a substantial decrease in left ventricles of failing hearts in all α1‐AR subtypes and in β1‐ and β2‐AR. However, the binding to β3‐AR was not changed. Our results suggest that α1‐AR changes might be part of a compensatory mechanism, by which the heart suffering from the HF tries to secure its function, and it could be hypothesized that ineffective β3‐AR regulation might be involved in development of HF. According to our knowledge, this is the first report about the β3‐AR binding in HF.
Activity of Oxytocinergic Neurons in the Supraoptic Nucleus under Stimulation of α2‐Adrenoceptors in Brattleboro Rats
Authors: Bundzikova Jana , Pirnik Zdeno , Mikkelsen Jens D. , Zelena Dora , Kiss Alexander
Published: November 2009

Vasopressin (AVP) deficient homozygous Brattleboro rats exhibit severe osmotic challenges due to waterless chronic hypernatremia and hyperosmolality. We investigated the effect of xylazine, an α2‐adrenoceptor agonist, on the activity of oxytocinergic (OXY) neurons in the supraoptic nucleus (SON) of homozygous (di/di), heterozygous (di/+), and control (+/+) rats. Ninety minutes after saline (0.1 mL/100 g b.w., i.p.) or xylazine injection (10 mg/kg, i.p.) rats were anaesthetized with pentobarbital (50 mg/kg i.p.) and sacrificed by transcardial perfusion with fixative. Activity of OXY neurons was evidenced by nuclear Fos protein immunoreactivity. Fos/OXY colabelings were analyzed on 40‐μm thick coronal sections using computerized light microscope. As expected, plasma osmolality and water intake revealed high heterogenity within the di/di group of rats. Fos expression in SON of di/di rats was correlated with osmolality of each rat. In saline‐treated rats, maximum activation of Fos reached around 4% in +/+, 20% in di/+ rats, and as much as 60% in di/di rats. Xylazine activated in SON about 70% of OXY‐ergic neurons in +/+, 60% in di/+ rats, and more than 80% in di/di rats. The present findings indicate that in spite of the high spontaneous activity of SON OXY‐ergic neurons due to the AVP deficiency in di/di rats, many of the silent OXY‐ergic neurons in the SON remained acceptable for α2‐adrenoceptor stimulation.
Regulation of Gene Expression of Catecholamine Biosynthetic Enzymes in Dopamine‐β‐Hydroxylase‐ and CRH‐Knockout Mice Exposed to Stress
Authors: Kvetňanský Richard , Krizanova Olga , Tillinger Andrej , Sabban Esther L. , Thomas Steven A. , Kubovcakova Lucia
Published: November 2009

Norepinephrine‐deficient mice harbor a disruption of the gene for dopamine‐β‐hydroxylase (DBH‐KO). Corticotropin‐releasing hormone knockout mice (CRH‐KO) have markedly reduced HPA activity. The aim of the present work was to study how deficiency of DBH and CRH would affect tyrosine hydroxylase (TH), DBH, and phenylethanolamine N‐methyltransferase (PNMT) gene expression and protein levels in the adrenal medulla (AM) and stellate ganglia (SG) of control and stressed mice. Both in AM and SG, single immobilization significantly increased TH and DBH mRNA and protein levels both in wild‐type (WT) and CRH‐KO mice. On the other hand, the stress‐triggered increase in PNMT mRNA and protein levels seen in WT mice was absent in CRH‐KO mice. DBH‐KO mice are more sensitive to stress but survive a single 2 h restraint stress in a tube. The increase in TH mRNA levels induced by restraint stress in WT was not observed in DBH‐KO mice. PNMT mRNA and especially PNMT protein levels were significantly elevated in AM of DBH‐KO mice. In SG of DBH‐KO mice, TH mRNA levels were not affected; however, PNMT gene expression was highly elevated. Thus, disruption of the DBH gene surprisingly blocks the stress‐induced elevation of TH mRNA levels in AM but increases PNMT gene expression in both AM and SG. Our data indicate that adrenergic signaling is required for stress‐induced increase in TH mRNA and that this signaling restrains stress‐induced increase in PNMT mRNA. They also confirm that the HPA system plays a crucial role in the stress‐induced regulation of PNMT gene expression.
Cross‐talk between Adrenal Medulla and Adrenal Cortex in Stress
Authors: Ehrhart‐Bornstein Monika , Bornstein Stefan R.
Published: November 2009

Response of the endocrine system to stress is characterized by the concomitant release of catecholamines from the adrenal medulla and of glucocorticoids from the adrenal cortex. Therefore, the activity of these two embryologically different endocrine organs that are united under one organ capsule has to be synchronized. The data summarized here indicate an involvement of intra‐adrenal interactions in this coordination of the body's response to stress.
Human Sympathetic Nerve Biology
Authors: Esler Murray , Eikelis Nina , Schlaich Markus , Lambert Gavin , Alvarenga Marlies , Kaye David , El‐Osta Assam , Guo Ling , Barton David , Pier Ciaran , Brenchley Celia , Dawood Tye , Jennings Garry , Lambert Elisabeth
Published: November 2009

Patients with panic disorder provide a clinical model of stress. On a “good day,” free from a panic attack, they show persistent stress‐related changes in sympathetic nerve biology, including abnormal sympathetic nerve single‐fiber firing (“salvos” of multiple firing within a cardiac cycle) and release of epinephrine as a cotransmitter. The coreleased epinephrine perhaps originates from in situ synthesis by phenylethanolamine N‐methyltransferase (PNMT). In searching for biological evidence that essential hypertension is caused by mental stress—a disputed proposition—we note parallels with panic disorder, which provides an explicit clinical model of stress: (1) There is clinical comorbidity; panic disorder prevalence is increased threefold in essential hypertension. (2) For both, epinephrine cotransmission is present in sympathetic nerves. (3) In panic disorder and essential hypertension, but not in health, single‐fiber sympathetic nerve firing salvos occur. (4) Tissue nerve growth factor is increased in both conditions (nerve growth factor is a stress reactant). (5) There is induction of PNMT in sympathetic nerves. Essential hypertension exhibits a further manifestation of mental stress: there is activation of noradrenergic brain stem neurons projecting to the hypothalamus and amygdala. These pathophysiological findings strongly support the view that chronic mental stress is important in the pathogenesis of essential hypertension. A hypothesis now under test is whether in both disorders, under prevailing conditions of ongoing stress, PNMT induced in sympathetic nerves acts as a DNA methylase, causing the norepinephrine transporter (NET) gene silencing that is present in both conditions. PNMT can have an intranuclear distribution, binding to DNA. We have demonstrated that the reduced neuronal noradrenaline reuptake present in both disorders does have an epigenetic mechanism, with demonstrable reduction in the abundance of the transporter protein, the NET gene silencing being associated with DNA binding by the methylation‐related inhibitory transcription factor MeCP2.
Effects of TWEAK (TNF Superfamily Member 12) on Differentiation, Metabolism, and Secretory Function of Human Primary Preadipocytes and Adipocytes
Authors: Tiller Gabriele , Fischer-Posovszky Pamela , Laumen Helmut , Finck Andreas , Skurk Thomas , Keuper Michaela , Brinkmann Ulrich , Wabitsch Martin , Link Dieter , Hauner Hans
Published: November 2009

Expansion of adipose tissue mass by hypertrophy and hyperplasia is the hallmark of obesity. An automated cDNA screen was established to identify secreted human proteins with an inhibitory effect on adipocyte differentiation and, thereby, a potential inhibitory effect on adipose tissue growth. A member of the TNF superfamily, TNF-like weak inducer of apoptosis (TWEAK; TNF superfamily 12) was identified by means of high-throughput screening with the lipophilic dye Nile Red as an inhibitor of murine adipocyte differentiation and, subsequently, also of human adipocyte differentiation. TWEAK inhibited lipid deposition in a dose-dependent manner without causing cytotoxic effects. This inhibitory action was mimicked by an agonistic antibody of the TWEAK receptor. The TWEAK receptor (fibroblast growth factor inducible 14; CD266) was expressed on human primary preadipocytes and mature adipocytes. Knockdown of TWEAK receptor by short-hairpin RNA abolished the inhibitory effect of TWEAK on cell differentiation, demonstrating that the effects of TWEAK are mediated by its specific receptor. Inhibition of differentiation was the result of interference at an early step of transcriptional activation as assessed by decreased peroxisome proliferator-activated receptor-γ, CCAAT enhancer-binding protein α (C/EBPα), and CCAAT enhancer-binding protein β (C/EBPβ) mRNA expression. In contrast to TNFα, basal and insulin-stimulated glucose uptake and lipolysis of terminally differentiated mature adipocytes and secretion of proinflammatory cytokines were not altered in the presence of TWEAK, and nuclear factor κ B activity was only weakly induced. We conclude from our findings that TWEAK and the corresponding agonistic antibody have the potential to prevent adipose tissue growth without adversely influencing central metabolic pathways or proinflammatory cytokine secretion in adipose tissue.
Wingless (Wnt)-3A Induces Trophoblast Migration and Matrix Metalloproteinase-2 Secretion through Canonical Wnt Signaling and Protein Kinase B/AKT Activation
Authors: Sonderegger Stefan , Haslinger Peter , Sabri Alia , Leisser Christina , Otten Jan V. , Fiala Christian , Knöfler Martin
Published: November 2009

Invasion of human trophoblasts is promoted through activation of wingless (Wnt) signaling, suggesting a role of the pathway in placental development and morphogenesis. However, details on the process such as involvement of canonical and/or noncanonical Wnt signaling cascades as well as their target genes are largely unknown. Hence, signal transduction via canonical Wnt signaling or phosphatidylinositide 3-kinase (PI3K)/AKT and their cross talk as well as trophoblast-specific protease expression were investigated in trophoblastic SGHPL-5 cells and primary extravillous trophoblasts purified from first-trimester placentas. Western blot analyses revealed that the recombinant Wnt ligand Wnt-3A increased phosphorylation of AKT and the downstream kinase glycogen synthase kinase (GSK)-3β as well as accumulation of activated, nuclear β-catenin. In accordance, luciferase expression of a canonical Wnt/TCF reporter and cell migration in first-trimester villous explant cultures and of SGHPL-5 cells were stimulated. Chemical inhibition of PI3K abolished Wnt-dependent phosphorylation of AKT and GSK-3β and trophoblast motility but did not affect appearance of activated β-catenin or Wnt/TCF reporter activity. In contrast, inhibition of the canonical pathway through soluble Dickkopf-1 did not influence AKT and GSK-3β phosphorylation but reduced Wnt reporter activity, accumulation of active β-catenin, and cell migration. Both inhibitors decreased Wnt-3A-induced secretion of pro- and active matrix metalloproteinase-2 from SGHPL-5 cells and pure EVT. The data suggest that Wnt-3A may activate canonical Wnt signaling and PI3K/AKT through distinct receptors. The two signaling cascades act independently in trophoblasts; however, both pathways promote Wnt-dependent migration and the release of matrix metalloproteinase-2, which has been identified as novel Wnt target in invasive trophoblasts.
Peroxisome Proliferator-Activated Receptor-α Control of Lipid and Glucose Metabolism in Human White Adipocytes
Authors: Ribet Carole , Montastier Emilie , Valle Carine , Bezaire Véronic , Mazzucotelli Anne , Mairal Aline , Viguerie Nathalie , Langin Dominique
Published: November 2009

This work aimed at characterizing the role of peroxisome proliferator-activated receptors (PPAR)α in human white adipocyte metabolism and at comparing PPARα and PPARγ actions in these cells. Primary cultures of human fat cells were treated with the PPARα agonist GW7647 or the PPARγ agonist rosiglitazone. Changes in gene expression were determined using DNA microrrays and quantitative RT-PCR. Western blot and metabolic studies were performed to identify the biological effects elicited by PPAR agonist treatments. GW7647 induced an up-regulation of β-oxidation gene expression and increased palmitate oxidation. Unexpectedly, glycolysis was strongly reduced at transcriptional and functional levels by GW7647 leading to a decrease in pyruvate and lactate production. Glucose oxidation was decreased. Triglyceride esterification and de novo lipogenesis were inhibited by the PPARα agonist. GW7647-induced alterations were abolished by a treatment with a PPARα antagonist. Small interfering RNA-mediated extinction of PPARα gene expression in hMADS adipocytes attenuated GW7647 induction of palmitate oxidation. Rosiglitazone had no major impact on glycolysis and β-oxidation. Altogether these results show that PPARα can selectively up-regulate β-oxidation and decrease glucose utilization in human white adipocytes.
Deletion of the Novel Oocyte-Enriched Gene, Gpr149, Leads to Increased Fertility in Mice
Authors: Edson Mark A. , Lin Yi-Nan , Matzuk Martin M.
Published: November 2009

Through in silico subtraction and microarray analysis, we identified mouse Gpr149, a novel, oocyte-enriched transcript that encodes a predicted orphan G-protein-coupled receptor (GPR). Phylogenetic analysis of GPR149 from fish to mammals suggests that it is widely conserved in vertebrates. By multitissue RT-PCR analysis, we found that Gpr149 is highly expressed in the ovary and also in the brain and the digestive tract at low levels. Gpr149 levels are low in newborn ovaries but increase throughout folliculogenesis. In the ovary, we found that granulosa cells did not express Gpr149, whereas germinal vesicle and meiosis II stage oocytes showed high levels of Gpr149 expression. After fertilization, Gpr149 expression declined, becoming undetectable by the two-cell stage. To study the function of GPR149 in oocyte growth and maturation, we generated Gpr149 null mice. Surprisingly, Gpr149 null mice are viable and have normal folliculogenesis, but demonstrate increased fertility, enhanced ovulation, increased oocyte Gdf9 mRNA levels, and increased levels of FSH receptor and cyclin D2 mRNA levels in granulosa cells. Thus, Gpr149 null mice are one of the few models with enhanced fertility, and GPR149 could be a target for small molecules to enhance fertility in the assisted reproductive technology clinic.
Central Insulin Signaling Is Attenuated by Long-Term Insulin Exposure via Insulin Receptor Substrate-1 Serine Phosphorylation, Proteasomal Degradation, and Lysosomal Insulin Receptor Degradation
Authors: Mayer Christopher M. , Belsham Denise D.
Published: November 2009

Central insulin signaling is critical for the prevention of insulin resistance. Hyperinsulinemia contributes to insulin resistance, but it is not yet clear whether neurons are subject to cellular insulin resistance. We used an immortalized, hypothalamic, clonal cell line, mHypoE-46, which exemplifies neuronal function and expresses the components of the insulin signaling pathway, to determine how hyperinsulinemia modifies neuronal function. Western blot analysis indicated that prolonged insulin treatment of mHypoE-46 cells attenuated insulin signaling through phospho-Akt. To understand the mechanisms involved, time-course analysis was performed. Insulin exposure for 4 and 8 h phosphorylated Akt and p70-S6 kinase (S6K1), whereas 8 and 24 h treatment decreased insulin receptor (IR) and IR substrate 1 (IRS-1) protein levels. Insulin phosphorylation of S6K1 correlated with IRS-1 ser1101 phosphorylation and the mTOR-S6K1 pathway inhibitor rapamycin prevented IRS-1 serine phosphorylation. The proteasomal inhibitor epoxomicin and the lysosomal pathway inhibitor 3-methyladenine prevented the degradation of IRS-1 and IR by insulin, respectively, and pretreatment with rapamycin, epoxomicin, or 3-methyladenine prevented attenuation of insulin signaling by long-term insulin exposure. Thus, a sustained elevation of insulin levels diminishes neuronal insulin signaling through mTOR-S6K1-mediated IRS-1 serine phosphorylation, proteasomal degradation of IRS-1 and lysosomal degradation of the IR.
Up-Regulation of Toll-Like Receptor 4/Nuclear Factor-κB Signaling Is Associated with Enhanced Adipogenesis and Insulin Resistance in Fetal Skeletal Muscle of Obese Sheep at Late Gestation
Authors: Yan Xu , Zhu Mei J. , Xu Wei , Tong Jun F. , Ford Stephen P. , Nathanielsz Peter W. , Du Min
Published: November 2009

Maternal obesity is increasing at an alarming rate. We previously showed that maternal obesity induces an inflammatory response and enhances adipogenesis in fetal skeletal muscle at midgestation. The objective of this study was to evaluate effects of maternal obesity on adipogenesis, inflammatory signaling, and insulin pathways at late gestation when ovine fetal skeletal muscle matures. Nonpregnant ewes were assigned to a control diet (Con, fed 100% of National Research Council nutrient recommendations, n = 6) or obesogenic diet (OB, fed 150% of National Research Council recommendations, n = 6) from 60 d before to 135 d after conception (term 148 d) when the fetal semitendenosus skeletal muscle was sampled. Expression of the adipogenic marker, peroxisome proliferator-activated receptor-γ, was increased in OB compared with Con fetal semitendenosus muscle, indicating up-regulation of adipogenesis. More intramuscular adipocytes were observed in OB muscle. Phosphorylation of inhibitor-κB kinase-α/β and nuclear factor-κB RelA/p65 were both increased in OB fetal muscle, indicating activation of nuclear factor-κB pathway. Phosphorylation of c-Jun N-terminal kinase and c-Jun (at Ser 63 and Ser 73) was also elevated. Toll-like receptor 4 expression was higher in OB than Con fetal muscle. Moreover, despite higher insulin concentrations in OB vs. Con fetal plasma (2.89 ± 0.53 vs. 1.06 ± 0.52 ng/ml; P < 0.05), phosphorylation of protein kinase B at Ser 473 was reduced, indicating insulin resistance. In conclusion, our data show maternal obesity-induced inflammatory signaling in late gestation fetal muscle, which correlates with increased im adipogenesis and insulin resistance, which may predispose offspring to later-life obesity and diabetes.
Minireview: Transgenerational Inheritance of the Stress Response: A New Frontier in Stress Research
Authors: Matthews Stephen G. , Phillips David I. W.
Published: November 2009

It is well established in animal models that the prenatal environment can have a major impact on stress axis function throughout life. These changes can predispose to various metabolic, cardiovascular, and neurobiological pathophysiologies. Emerging evidence indicates that the same programming effects occur in humans. It is now becoming clear that the pathophysiological effects are not confined to the first-generation offspring and that there is transgenerational memory of fetal experience that can extend across multiple generations. The complex mechanisms by which transgenerational transmission of stress responsiveness occur are rapidly becoming a focus of investigation. Understanding these fundamental biological processes will allow for development of intervention strategies that prevent or reverse adverse programming of the stress response.
Imitating the Affects of Beasts: Interest and Inhumanity in Spinoza
Authors: Montag Warren
Published: November 2009

Is Spinoza a theoretician of self-interest? Historians of philosophy have arrived at diametrically opposed answers to this question. His introduction of the term conatus, meaning endeavor or striving, in part 3 of the Ethics, together with his assertion that when human beings most seek that which is useful to them individually they are most useful to each other, might appear, as A. O. Hirschman argued, to render him both a follower of the Stoics and a predecessor of Mandeville and Smith. This essay takes the opposite view, arguing that Spinoza is above all the theoretician of the self-destruction and self-negation that societies of servitude require. The author calls these tendencies the inhumanization of politics, the forms in which the human is systematically subjected through affective imitation to what is incompatible with human existence.
In vivo roles for Arp2/3 in cortical actin organization during C. elegans gastrulation
Authors: Roh-Johnson Minna , Goldstein Bob
Published: November 2009

The Arp2/3 complex is important for morphogenesis in various developmental systems, but specific in vivo roles for this complex in cells that move during morphogenesis are not well understood. We have examined cellular roles for Arp2/3 in the Caenorhabditis elegans embryo. In C. elegans, the first morphogenetic movement, gastrulation, is initiated by the internalization of two endodermal precursor cells. These cells undergo a myosin-dependent apical constriction, pulling a ring of six neighboring cells into a gap left behind on the ventral surface of the embryo. In agreement with a previous report, we found that in Arp2/3-depleted C. elegans embryos, membrane blebs form and the endodermal precursor cells fail to fully internalize. We show that these cells are normal with respect to several key requirements for gastrulation: cell cycle timing, cell fate, apicobasal cell polarity and apical accumulation and activation of myosin-II. To further understand the function of Arp2/3 in gastrulation, we examined F-actin dynamics in wild-type embryos. We found that three of the six neighboring cells extend short, dynamic F-actin-rich processes at their apical borders with the internalizing cells. These processes failed to form in embryos that were depleted of Arp2/3 or the apical protein PAR-3. Our results identify an in vivo role for Arp2/3 in the formation of subcellular structures during morphogenesis. The results also suggest a new layer to the model of C. elegans gastrulation: in addition to apical constriction, internalization of the endoderm might involve dynamic Arp2/3-dependent F-actin-rich extensions on one side of a ring of cells.
Crucial role of phosphatidylinositol 4-kinase IIIα in development of zebrafish pectoral fin is linked to phosphoinositide 3-kinase and FGF signaling
Authors: Ma Hui , Blake Trevor , Chitnis Ajay , Liu Paul , Balla Tamas
Published: November 2009

Phosphatidylinositol 4-kinases (PI4Ks) catalyze the first committed step in the synthesis of phosphoinositides, important lipid regulators of signaling and trafficking pathways. Here we cloned Pik4a, one of the zebrafish PI4K enzymes, and studied its role(s) in vertebrate development using morpholino oligonucleotide-based gene silencing in zebrafish. Downregulation of Pik4a led to multiple developmental abnormalities, affecting the brain, heart, trunk and most prominently causing loss of pectoral fins. Strikingly similar defects were caused by treatment of the developing embryos with the phosphoinositide 3-kinase (PI3K) inhibitor, LY294002. To investigate the cause of the pectoral fin developmental defect, we focused on fibroblast growth factor (FGF) signaling pathways because vertebrate limb development requires the concerted action of a series of FGF ligands. Using in situ hybridization, the pectoral fin defect was traced to disruption of the early FGF signaling loops that are crucial for the establishment of the sharp signaling center formed by the apical ectodermal ridge and the underlying mesenchyme. This, in turn caused a prominent loss of the induction of one of the mitogen-activated protein kinase (MAPK) phosphatases, Mkp3, an essential intermediate in vertebrate limb development. These changes were associated with impaired proliferation in the developing fin bud due to a loss of balance between the MAPK and PI3K branch of FGF-initiated signals. Our results identify Pik4a as an upstream partner of PI3Ks in the signaling cascade orchestrated by FGF receptors with a prominent role in forelimb development.
Impact of energy efficiency on computer numerically controlled machining
Authors: Anderberg S E , Kara S , Beno T
Published: November 2009

Increasing environmental demands from governmental bodies and customers stress the importance of companies improving their environmental performance. The research presented here shows that productivity and cost efficiency improvements can be achieved alongside energy savings in a computer numerically controlled machining environment. This improves the profitability of the companies, but also leads them towards more sustainable and environmentally aware manufacturing; the relationship between machining parameters, machining costs, and energy consumption is evaluated. From this perspective, it is important that production planners etc. understand the methodological possibilities for improvements in cost and energy efficiency. The current research is based on a machining cost model and experiments where energy consumption and tool wear were monitored.
Prediction of drilling-induced damage in unidirectional glass-fibre-reinforced plastic laminates using an artificial neural network
Authors: Mishra R , Malik J , Singh I
Published: November 2009

Drilling-induced damage is a serious problem in laminated composite materials. The research efforts worldwide have been focused on minimization of this damage. A number of methodologies have been adopted for this purpose. The present research effort is aimed towards developing a predictive tool for calculating the likely damage before actual drilling commences, and thereby reducing its severity. The artificial neural network topology has been adopted as a predictive tool. The spindle speed, feed rate, drill diameter, and drill point geometry have been used as the input parameters. The drilling-induced damage was the output. The experimental data for drilling of unidirectional glass-fibre-reinforced plastic composite laminates were used for training and testing the model. The results of the predictive model have been found to be in good agreement with the test data.
Determination of local thrust coefficients of an aerospace component with the fibre Bragg grating array
Authors: Kim K , Kwon W C , Yoon J Y , Lee J M , Hwang Y
Published: November 2009

This paper describes a novel thrust measurement on a rotating wooden blade using the fibre Bragg grating (FBG) array. Five FBG sensors created in a single fibre were installed on the surface of a blade along the radial direction and one FBG sensor was placed in the middle of a rotor shaft. Measurements were performed with two wooden blades under loading in hover and with a dynamic FBG interrogator maintaining six channels for analysing optic signals. Distributed forces normal to the surface of a blade were determined with the assumptions of the Euler—Bernoulli beam theory, and local thrust coefficients were calculated with blade element momentum theory (BEMT). Direct comparison between measured local thrust coefficient values and calculated ones was performed and good agreement was obtained on the local thrust coefficient versus the dimensionless radial distance chart. Additionally, the deflection shape of the blade in the flapwise direction was determined at various rotational speeds. Theoretical and experimental procedures described in this article offer an effective and improved method for the measurement and analysis of thrust coefficients for blade design as well as maintenance.
Laser-assisted Direct Write for aerospace applications
Authors: Sato T , Fearon E , Curran C , Watkins K G , Dearden G , Eckford D
Published: November 2009

Particulate silver inks are finding increasing use in the direct manufacture of electronic circuitry, for example in Direct Write (DW). Laser curing of these inks as opposed to traditional oven-curing methods is advantageous for reasons of speed and the protection of nearby heat-sensitive components, and particularly for the construction of circuitry on conformal surfaces and ‘on the fly’ modification or production.

To fully appreciate the benefits provided by using a laser-based process, it is important that the final-cured component has electrical and physical properties that are comparable to, if not better than, those that are produced using the conventional technique. Currently, in some single pass curing techniques, cavities are formed in the track due to rapid boiling off of solvents that leads to a higher resistivity and surface roughness than that of oven-cured tracks. While mass loss is similar, overall density is decreased through porosity of the tracks. By going from a single pass curing process to a multiple pass process and controlling the power regime, it is possible to improve the track quality by allowing the gradual release of solvent vapour from the ink and achieve lower resistivities than that of oven-cured samples and a surface roughness that is comparable. This incremental process improves settling of the silver particulates leading to a resistance of the cured inks that is lower than that achievable through oven curing. This creates the possibility of fabricating a surface-mounted or -embedded damage sensor without thermally affecting the substrate.

The effect of substrate type thermal conductivity on the laser curing speed of silver inks is investigated. It was found that a noticeable factor in determining the resulting resistance of the conductive track was the substrate thermal conductivity. Both CO2 and Er:YAG laser radiation was investigated. Although CO2 lasers are in widespread use, the increasing demand for portability and ease of integration into existing DW systems favours fibre delivered laser radiation such as that from an Er:YAG fibre laser. This source was also investigated here to determine if fibre delivered processing could be achieved.