Phosphorylation of CARMA1: the link(er) to NF-kappaB activation.

Antigen receptor-induced NF-kappaB activation depends on receptor-proximal and -distal signaling events. Two papers in this issue of Immunity demonstrate that PKC-dependent phosphorylation of CARMA1 is the critical molecular link that controls the activation of the IKK signalosome and NF-kappaB.

Role of MT1-MMP in cancer dissemination : insights from the real time imaging of tumors

Abstract : Matrix metalloproteinases (MMPs) are thought to play a major role in the tumor dissemination process as they degrade all components of the extracellular matrix. However, failure of clinical trials testing broad MMP inhibitors in cancer led to the consensus that a better understanding of the MMP biology was required. Using intravital multiphoton laser scanning microscopy, we developed an in vivo model to observe tumor dissemination and extracellular matrix remodeling in real time. We show that the matrix-modifying hormone relaxin increases tumor associated fibroblast interaction with collagen fibers by inducing integrin beta-1 expression. This causes changes in the collagen network that are mediated by MMP-8 and MT1-MMP. Also, we show that MMP-mediated collagen remodeling in vivo requires a direct contact between stationary tumor associated fibroblasts (TAFs) and collagen fibers. As MMPs are expressed in the tumor and stromal compartment of breast cancers we determined the importance of Membrane-type 1 MMP (MT1-MMP) from each compartment for cancer progression. We find that tumor-MT1-MMP promotes the invasion of the blood vasculature and blood-borne metastasis in vivo by enhancing tumor cell migration and endothelial basement membrane degradation. Interestingly, stromal-MT1-MMP cannot compensate for the lack of tumor-MT1-MMP but promotes peritumor collagen I remodeling. Thus, the function of MT1-MMP is context dependent and we identify the different but complementary roles of tumor and stromal MT1-MMP for tumor dissemination. Finally, we translate our preclinical findings in to human breast cancer samples. We show that tumor-MT1-MMP expression correlates with tumor invasion of the blood vasculature in ER-PR-HER2- breast cancers and that MT1-MMP expression increases with cancer progression. MT1-MMP could thus represent an interesting therapeutic target for the prevention of blood vasculature invasion in these tumors. Resumé : Les matrix metalloproteinases (MMPs) semblent jouer un rôle majeur pour la dissémination tumorale en raison de leur capacité à dégrader l'ensemble des composants de la matrice extracellulaire (MEC). Néanmoins, les résultats décevants des études cliniques testant les inhibiteurs des MMP ont conduit à la notion qu'une compréhension plus précise de la biologie des MMP était requise. Dans ce travail de thèse, nous avons développé un modèle murin qui permet d'observer simultanément la dissémination tumorale ainsi que les modifications de la MEC en temps réel. Nous démontrons que le traitement de tumeurs par l'hormone relaxin augmente l'interaction des fibroblastes tumoraux avec les fibres de collagène via l'intégrine beta-1. Nous montrons que cette interaction favorise et est nécessaire à la dégradation des fibres de collagène par MMP-8 et MT1-MMP. Ensuite, étant donné que les MMPs sont exprimées dans les cellules tumorales et stromales des cancers du sein, nous nous sommes intéressés au rôle de la MMP membranaire type 1 (MT1-MMP) exprimée dans chacun de ces compartiments. Nous démontrons que MT1-MMP dérivant des cellules tumorales favorise leur invasion dans les vaisseaux sanguins par la dégradation de la membrane basale vasculaire. De manière inattendue, nous montrons que l'expression de MT1-MMP par le compartiment stromal ne peut compenser le manque de MT1-MMP dans le compartiment tumoral. Néanmoins, nos résultats prouvent que MT1-MMP dérivant du compartiment stromal est impliqué dans la dégradation de collagène peritumorale. La fonction de la protéine MT1-MMP varie donc selon le compartiment tumoral d'origine. Finalement, nous avons testé nos résultats pré cliniques chez l'humain. Dans des biopsies de cancer du sein nous montrons une corrélation entre l'expression de MT1-MMP dans les cellules tumorales et l'invasion de vaisseaux sanguins par des tumeurs ER-PR-HER2-. MT1-MMP pourrait donc être une cible intéressante pour la prévention de dissémination vasculaire de ces tumeurs

Rockfall monitoring by Terrestrial Laser Scanning ¿ case study of the basaltic rock face at Castellfollit de la Roca (Catalonia, Spain)

This case study deals with a rock face monitoring in urban areas using a Terrestrial Laser Scanner. The pilot study area is an almost vertical, fifty meter high cliff, on top of which the village of Castellfollit de la Roca is located. Rockfall activity is currently causing a retreat of the rock face, which may endanger the houses located at its edge. TLS datasets consist of high density 3-D point clouds acquired from five stations, nine times in a time span of 22 months (from March 2006 to January 2008). The change detection, i.e. rockfalls, was performed through a sequential comparison of datasets. Two types of mass movement were detected in the monitoring period: (a) detachment of single basaltic columns, with magnitudes below 1.5 m3 and (b) detachment of groups of columns, with magnitudes of 1.5 to 150 m3. Furthermore, the historical record revealed (c) the occurrence of slab failures with magnitudes higher than 150 m3. Displacements of a likely slab failure were measured, suggesting an apparent stationary stage. Even failures are clearly episodic, our results, together with the study of the historical record, enabled us to estimate a mean detachment of material from 46 to 91.5 m3 year¿1. The application of TLS considerably improved our understanding of rockfall phenomena in the study area.

Résultats à deux ans de la trabéculoplastie sélective au laser dans le glaucome à angle ouvert et l'hypertonie oculaire

BACKGROUND: Selective laser trabeculoplasty (SLT) is a relatively new treatment strategy for the treatment of glaucoma. Its principle is similar to that of argon laser trabeculoplasty (ALT), but may lead to less damage to the trabecular meshwork. METHODS: We assessed the 2-year efficacy of SLT in a noncomparative consecutive case series. Any adult patient either suspected of having glaucoma or with open-angle glaucoma, whose treatment was judged insufficient to reach target intraocular pressure (IOP), could be recruited. IOP and number of glaucoma treatments were recorded over 2 years after the procedure. RESULTS: Our sample consisted of 44 consecutive eyes of 26 patients, aged 69+/-8 years. Eyes were treated initially on the lower 180 degrees . Three of them were retreated after 15 days on the upper 180. Fourteen eyes had ocular hypertension, 17 primary open-angle/normal-tension glaucoma, 11 pseudoexfoliation (PEX) glaucoma, and two pigmentary glaucoma. Thirty-six eyes had previously been treated and continued to be treated with topical anti-glaucoma medication, ten had had prior ALT, nine iridotomy, and 12 filtering surgery. The 2-year-follow up could not be completed for eight eyes because they needed filtering surgery. In the remaining 36 eyes, IOP decreased by a mean of 17.2%, 3.3 mmHg, (19.2+/-4.7 to 15+/-3.6 mmHg) after 2 years (p<0.001). As a secondary outcome, the number of glaucoma treatments decreased from 1.44 to 1.36 drops/patient. Other results according to subgroups of patients are analyzed: the greatest IOP decrease occurred in eyes that had never been treated with anti-glaucoma medication or with PEX glaucoma. SLT was probably valuable in a few eyes after filtering surgery; however, the statistical power of the study was not strong enough to draw a firm conclusion. When expressed in survival curves after 2 years, however, only 48% and 41% of eyes experienced a decrease of more than 3 mmHg or more than 20% of preoperative intraocular pressure, respectively. CONCLUSION: SLT decreases IOP somewhat for at least 2 years without an increase in topical glaucoma treatment. However, it cannot totally replace topical glaucoma treatment. In the future, patient selection should be improved to decrease the cost/effectiveness ratio.

Fetoscopic laser therapy for twin-twin transfusion syndrome: beyond current gestational age limits.

Both "early" (< 16 weeks' gestation) and "late" (> 26 weeks' gestation) presentations of twin-twin transfusion syndrome (TTTS) are rare and challenging complications of monochorionic/diamniotic twin pregnancies. Growing evidence suggests that fetoscopic laser therapy for both "early" and "late" TTTS is feasible, safe, and yields similar outcomes to cases treated between 16 and 26 weeks' gestation. We suggest reevaluation of conventional gestational age guidelines for laser therapy for TTTS.

Evaluation of spatial distribution of flatfish by laser scannig

Peer reviewed

Determinação do volume de madeira em povoamento de eucalipto por escâner a laser aerotransportado

O objetivo deste trabalho foi avaliar a possibilidade de se estimar o diâmetro à altura do peito (DAP) com os dados de altura e de número de árvores derivados do escâner a laser aerotransportado (LiDAR, "light detection and ranging"), e determinar o volume de madeira de talhão de Eucalyptus sp. a partir dessas variáveis. O número total de árvores detectadas foi obtido com uso da filtragem de máxima local. A altura de plantas estimada pelo LiDAR apresentou tendência não significativa à subestimativa. A estimativa do DAP foi coerente com os valores encontrados no inventário florestal; porém, também mostrou tendência à subestimativa, em razão do comportamento observado quanto à altura. A variável número de fustes apresentou valores próximos aos observados nas parcelas do inventário. O LiDAR subestimou o volume total de madeira do talhão em 11,4%, em comparação ao volume posto na fábrica. A tendência de subestimação da altura das árvores (em média, cerca de 5%) impactou a estimativa do volume individual de árvores e, consequentemente, a do volume do talhão. No entanto, é possível gerar equações de regressão que estimam o DAP com boa precisão, a partir de dados de altura de plantas obtidos pelo LiDAR. O modelo parabólico é o que possibilita as melhores estimativas da produção volumétrica dos talhões de eucalipto.

Laser-MAG-hybridihitsauksen käyttöönotto laivan rungon valmistuksessa

Perinteisten kaarihitsausmenetelmien suhteellisen suuri lämmöntuonti aiheuttaa huomattavia muodonmuutoksia laivan rungon valmistusprosessin alkuvaiheessa. Muodonmuutosten seurauksena rakenteiden mitta- ja muototarkkuus heikkenee, mikä lisää oikaisu- ja sovitustyötä myöhemmissä työvaiheissa. Hitsausmuodonmuutoksia voidaan vähentää siirtymällä käyttämään laser-MAG-hybridihitsausta, jossa lämmöntuonti on merkittävästi pienempi kuin kaarihitsauksessa. Näin kyetään oleellisesti leikkaamaan oikaisu- ja sovitustyöstä syntyviä kustannuksia. Tämän diplomityön tavoitteena oli kehittää tuotantovalmiiksi kuitulaser- ja MAG-hitsauksen yhdistelmäprosessi Aker Yards Oy:n Turun telakalla loppuvuoden 2006 aikana. Hitsauslaitteiston asennus oli valmistunut kesäkuussa 2006, minkä jälkeen aloitettiin luokituslaitoksen hyväksymän koeohjelman hitsaukset. Käyttöönotto suunnitelmaan sisältyvä koehitsausohjelma oli laadittu Det Norske Veritaksen julkaisemaa ohjetta (Guidelines no. 19) mukaillen. Ensimmäiseksi määritettiin hitsauskokeiden avulla prosessille laadun ja tehokkuuden suhteen optimaalinen railogeometria. Seuraavaksi optimoitiin prosessin hitsausparametrit 6 mm:n aineenpaksuudelle hyödyntäen Taguchi-koesuunnittelumenetelmää. Tämän jälkeen optimiparametreilla hitsattiin koekappale väsytyskokeisiin, jotka suoritettiin Teknillisen korkeakoulun laivalaboratoriossa. Väsytyskoetulokset täyttivät luokituslaitoksen vaatimukset. Myös hitsauksen menetelmäkoe suoritettiin hyväksytetysti. Viimeinen koeohjelman mukainen hitsauskoesarja tehtiin prosessiparametrien sallittujen vaihtelurajojen määrittämiseksi. Diplomityön tavoite täyttyi joulukuussa 2006, jolloin 'laivan kansipaneeli hitsattiin ensimmäistä kertaa uudella hitsausprosessilla. Hitsauksen laatu korreloi hyvin menetelmäkokeen tulosten kanssa ¿ hitsit olivat tasalaatuisia ja ne täyttivät B-luokan vaatimukset.

CO2 Laser Cutting of Particleboard, HDF and MDF

The purpose of this thesis is to reveal how the laser cutting parameters influence lasercutting of particleboard, HDF and MDF. The literature review introduces the basic principle of CO2 laser, CO2 laser equipment and its usage in cutting of wood-based materials. The experimental part focuses on the discussion and analysis ofthe test data and attempts to draw conclusions on the influence of various parameters, including laser power, focal length of the lens and cutting gas, on the cutting speed and kerf quality. The tested materials include various thicknesses of particleboard, HDF and MDF samples. A TRUMPF TLF2700 HQ laser equipment was used for the experiments. To obtain valid data, the test samples must be completely cut through without any bonding of wood fibre. The maximum cutting speed is linear dependent on the laser power in thecondition that the other parameters are constant. For each thickness of a specific material type, there is a minimum laser power for cutting. Normally, the topand bottom kerf widths increase with the enhancement of laser power. There may be a critical laser power which can generate the minimum cross-sectional kerf width. Lens of larger focal length may achieve higher cutting speed. As the focal length becomes larger, the top kerf width tends to increase while the bottom andcross-sectional kerf widths to the opposite. Of all cutting gases, oxygen can help achieve higher cutting speed. The gas pressure of nitrogen does not seem to have strong influence on the cutting result. Generally, 2 bar air is more preferable for higher cutting speed. For particleboard and MDF samples of larger thickness than 12 mm, 2 bar argon can be used to reach remarkably higher cutting speed than the 5 bar. Generally, the 190.5 mm lens can produce smallest total kerf width. The kerf sides of thicker samples are darker than the thinner ones. The sample darkness tends to be lower as laser power increased. 63.5 mm lens seemed tocause more darkness than other lens. 5 bar cutting gases can produce less dark side kerfs than 2 bar ones. Oxygen normally causes darker kerfs than other gases. No distinct differences were found between nitrogen and argon.

Can resistance to trastuzumab be reversed by endocrine therapy? Combination of trastuzumab and letrozole after resistance to sequential trastuzumab and aromatase inhibitor monotherapies in patients with ER-positive, HER-2 positive, advanced breast cancer: a proof-of-concept trial (SAKK 23/03)

Introduction: Trastuzumab (T) is a cornerstone in the treatment of patients with HER2-overexpressing advanced breast cancer and development of resistance to T is a major therapeutic problem. HER-2 is part of a highly interactive signaling network that may impair efficacy of endocrine therapy. A sequential treatment design was chosen in this trial to ensure complete resistance to single agent therapy before receiving both a non-steroidal aromatase inhibitor (AI) and T. Any kind of clinical activity with combined treatment of AI and T after progression of single agent treatments could indicate restoration of sensitivity as a consequence of cross-talking and networking between both pathways. Methods: Key eligibility criteria included postmenopausal patients (pts.) with advanced, measurable, HER-2 positive (assessed by FISH, ratio (≥2)), HR positive disease and progression on prior treatment with a non-steroidal AI, e.g. letrozole or anastrozole, either in an adjuvant or advanced setting. Pts. received standard dose T monotherapy either weekly or three-weekly in step 1 and upon disease progression, continued T in combination with letrozole in step 2. The primary endpoint was clinical benefit response (CBR: CR, PR or SD for at least 24 weeks (+/- 1 week) according to RECIST) in step 2. Results: Thirteen pts. were enrolled in five centers in Switzerland. In step 1, six pts. (46%) achieved CBR. Median time to progression (TTP) was 161 days (Range: 50 - 627). Based on data collected until the end of May 2010, CBR was observed in seven out of the eleven evaluable pts. (64%) in step 2, including one pt. with partial response. Four of the seven pts. within step 2 that achieved CBR also had CBR in step 1. Seven out of eleven pts. have documented tumor progression during step 2 treatment. Median TTP for all eleven pts. was 184 days (range 61 - 471). Mean time on study treatment (TTP in step 1 plus TTP in step 2) for pts. reaching step 2 was 380 days (range 174 - 864). Adverse events were generally mild. Conclusion: Results of this proof-of-principle trial suggest that complete resistance to both AI and T can be overcome in a proportion of pts. by combined treatment of AI and T, as all pts. served as their own control. Our results appear promising for a new treatment strategy which offers a chemotherapy-free and well-tolerated option for at least a subset of the pts. with HR positive, HER-2 positive breast cancer. Further trials will need to corroborate this finding.

Morbillivirus glycoprotein expression induces ER stress, alters Ca2+ homeostasis and results in the release of vasostatin.

Although the pathology of Morbillivirus in the central nervous system (CNS) is well described, the molecular basis of neurodegenerative events still remains poorly understood. As a model to explore Morbillivirus-mediated CNS dysfunctions, we used canine distemper virus (CDV) that we inoculated into two different cell systems: a monkey cell line (Vero) and rat primary hippocampal neurons. Importantly, the recombinant CDV used in these studies not only efficiently infects both cell types but recapitulates the uncommon, non-cytolytic cell-to-cell spread mediated by virulent CDVs in brain of dogs. Here, we demonstrated that both CDV surface glycoproteins (F and H) markedly accumulated in the endoplasmic reticulum (ER). This accumulation triggered an ER stress, characterized by increased expression of the ER resident chaperon calnexin and the proapoptotic transcription factor CHOP/GADD 153. The expression of calreticulin (CRT), another ER resident chaperon critically involved in the response to misfolded proteins and in Ca(2+) homeostasis, was also upregulated. Transient expression of recombinant CDV F and H surface glycoproteins in Vero cells and primary hippocampal neurons further confirmed a correlation between their accumulation in the ER, CRT upregulation, ER stress and disruption of ER Ca(2+) homeostasis. Furthermore, CDV infection induced CRT fragmentation with re-localisation of a CRT amino-terminal fragment, also known as vasostatin, on the surface of infected and neighbouring non-infected cells. Altogether, these results suggest that ER stress, CRT fragmentation and re-localization on the cell surface may contribute to cytotoxic effects and ensuing cell dysfunctions triggered by Morbillivirus, a mechanism that might potentially be relevant for other neurotropic viruses.

Visual measurement of laser-arc hybrid welding

Tässä työssä raportoidaan hybridihitsauksesta otettujen suurnopeuskuvasarjojen automaattisen analyysijärjestelmän kehittäminen.Järjestelmän tarkoitus oli tuottaa tietoa, joka avustaisi analysoijaa arvioimaan kuvatun hitsausprosessin laatua. Tutkimus keskittyi valokaaren taajuuden säännöllisyyden ja lisäainepisaroiden lentosuuntien mittaamiseen. Valokaaria havaittiin kuvasarjoista sumean c-means-klusterointimenetelmän avullaja perättäisten valokaarien välistä aikaväliä käytettiin valokaaren taajuuden säännöllisyyden mittarina. Pisaroita paikannettiin menetelmällä, jossa yhdistyi pääkomponenttianalyysi ja tukivektoriluokitin. Kalman-suodinta käytettiin tuottamaan arvioita pisaroiden lentosuunnista ja nopeuksista. Lentosuunnanmääritysmenetelmä luokitteli pisarat niiden arvioitujen lentosuuntien perusteella. Järjestelmän kehittämiseen käytettävissä olleet kuvasarjat poikkesivat merkittävästi toisistaan kuvanlaadun ja pisaroiden ulkomuodon osalta, johtuen eroista kuvaus- ja hitsausprosesseissa. Analyysijärjestelmä kehitettiin toimimaan pienellä osajoukolla kuvasarjoja, joissa oli tietynlainen kuvaus- ja hitsausprosessi ja joiden kuvanlaatu ja pisaroiden ulkomuoto olivat samankaltaisia, mutta järjestelmää testattiin myös osajoukon ulkopuolisilla kuvasarjoilla. Testitulokset osoittivat, että lentosuunnanmääritystarkkuus oli kohtuullisen suuri osajoukonsisällä ja pieni muissa kuvasarjoissa. Valokaaren taajuuden säännöllisyyden määritys oli tarkka useammassa kuvasarjassa.

Acyl-CoA synthetase 3 promotes lipid droplet biogenesis in ER microdomains.

Control of lipid droplet (LD) nucleation and copy number are critical, yet poorly understood, processes. We use model peptides that shift from the endoplasmic reticulum (ER) to LDs in response to fatty acids to characterize the initial steps of LD formation occurring in lipid-starved cells. Initially, arriving lipids are rapidly packed in LDs that are resistant to starvation (pre-LDs). Pre-LDs are restricted ER microdomains with a stable core of neutral lipids. Subsequently, a first round of"emerging" LDs is nucleated, providing additional lipid storage capacity. Finally, in proportion to lipid concentration, new rounds of LDs progressively assemble. Confocal microscopy and electron tomography suggest that emerging LDs are nucleated in a limited number of ER microdomains after a synchronized stepwise process of protein gathering, lipid packaging, and recognition by Plin3 and Plin2. A comparative analysis demonstrates that the acyl-CoA synthetase 3 is recruited early to the assembly sites, where it is required for efficient LD nucleation and lipid storage.

Acyl-CoA synthetase 3 promotes lipid droplet biogenesis in ER microdomains.

Control of lipid droplet (LD) nucleation and copy number are critical, yet poorly understood, processes. We use model peptides that shift from the endoplasmic reticulum (ER) to LDs in response to fatty acids to characterize the initial steps of LD formation occurring in lipid-starved cells. Initially, arriving lipids are rapidly packed in LDs that are resistant to starvation (pre-LDs). Pre-LDs are restricted ER microdomains with a stable core of neutral lipids. Subsequently, a first round of"emerging" LDs is nucleated, providing additional lipid storage capacity. Finally, in proportion to lipid concentration, new rounds of LDs progressively assemble. Confocal microscopy and electron tomography suggest that emerging LDs are nucleated in a limited number of ER microdomains after a synchronized stepwise process of protein gathering, lipid packaging, and recognition by Plin3 and Plin2. A comparative analysis demonstrates that the acyl-CoA synthetase 3 is recruited early to the assembly sites, where it is required for efficient LD nucleation and lipid storage.