New insights into small molecules inhibitors and protein-protein interactions of VirB8 : a critical conserved component of the type IV secretion system.

Les systèmes bactériens de sécrétion de type IV (T4SS) sont constitués d’un ensemble de 8 à 12 protéines conservées. Ces dernières sont utilisées lors de la translocation de protéines, la translocation de complexes ADN-protéines mais aussi pour le transport de ces derniers au travers de la membrane cellulaire. Les T4SS, en tant que facteurs de virulence pour beaucoup de pathogènes comme Brucella suis, sont donc d’excellents modèles cibles pour le développement de médicaments d’antivirulence. Ces médicaments, en privant le pathogène de son facteur essentiel de virulence : le T4SS, constituent une alternative ou encore une amélioration des traitements antibiotiques utilisés actuellement. VirB8, un facteur d’assemblage conservé dans le T4SS, forme des dimères qui sont importants pour la fonction des T4SS dans ces pathogènes. De par ses interactions multiples, VirB8 est un excellent modèle pour l’analyse des facteurs d’assemblage mais aussi en tant que cible de médicaments qui empêcheraient son interaction avec d’autres protéines et qui, in fine, désarmeraient les bactéries en les privant de leur fonctions essentielles de virulence. À ce jour, nous savons qu’il existe un équilibre monomère-dimère et un processus d’homodimerization de VirB8 dont l’importance est vitale pour la fonctionnement biologique des T4SSs. En se basant sur des essais quantitatifs d’interaction, nous avons identifié (i) des sites potentiels d’interaction avec d’autres protéines VirB du T4SS mais aussi (ii) isolé des petites molécules inhibitrices afin de tester la fonction protéique de VirB8. Afin de déterminer les acides aminés importants pour l’hétérodimérization de VirB8 avec VirB10, nous avons effectué des expériences de mutagenèse aléatoire, de phage display et d’arrimage moléculaire in silico. Ces expériences ont démontré l’importance de trois acides aminés localisés sur le feuillet β : R160, S162, T164 et I165. Ces derniers seraient importants pour l’association de VirB8 avec VirB10 étant donné que leur mutagenèse entraine une diminution de la formation du complexe VirB8-VirB10. L’objectif actuel de notre projet de recherche est de pouvoir mieux comprendre mais aussi d’évaluer le rôle de VirB8 dans l’assemblage du T4SS. Grace à un méthode de criblage adaptée à partir de la structure de VirB8, nous avons pu identifié une petite molécule inhibitrice BAR-068, qui aurait un rôle prometteur dans l’inhibition du T4SS. Nous avons utilisé la spectroscopie par fluorescence, l’essai à deux hybrides, le cross-linking et la cristallographie afin de déterminer le mécanisme d'interaction existant entre VirB8 et BAR-068. Ces travaux pourraient permettre de nombreuses avancées, notamment en termes de compréhension des mécanismes d’inhibition du T4SS. Notre objectif ultime est de pouvoir caractériser la séquence d’évènements essentiels à l’assemblage et au fonctionnement du T4SS. De manière globale, notre projet de recherche permettrait de révéler les grands principes d’assemblage des protéines membranaires, les processus de sécrétion de protéines chez les bactéries mais aussi de proposer une nouvelle stratégie lors du développement de drogues antimicrobiennes.

Sympathetic Stimulation and Hypertension in the Pyridoxine-Deficient Adult Rat

Pyridoxal phosphate is the coenzyme of various decarboxylases involved in the formation of monoamine neurotransmitters such as y-aminobutyric acid , serotonin , dopamine, and norepinephrine . Adult male Sprague-Dawley rats placed on a pyridoxine -deficient diet for 8 weeks showed significant hypertension compared with pyridoxine -supplemented controls . Hypothalamic contents of pyridoxal phosphate , y-aminobutyric acid, and serotonin in the pyridoxine - deficient rats were significantly lower than those in pyridoxine -supplemented controls . Hypertension was associated with sympathetic stimulation . Treatment of pyridoxine-deficient rats with a single dose of pyridoxine (10 mg/kg body weight) reversed the blood pressure to normal levels within 24 hours, with concomitant restorations of hypothalamic serotonin and y-aminobutyric acid as well as the return of plasma norepinephrine and epinephrine to normal levels . Also, pyridoxine treatment reversed the hypothalamic hypothyroidism observed in pyridoxine -deficient rats . These results indicate an association between pyridoxine deficiency and sympathetic stimulation leading to hypertension.

Three Component Mannich Reaction and 1,5-Benzodiazepine Synthesis Catalyzed by a Tetranitrile-Silver Complex

This manuscript describes the first example of silver ion complex of a dendritic tetranitrile ligand catalyzed one-pot three component Mannich reaction and 1,5-benzodiazepine synthesis. The catalyst can be separated from the products by a change in the solvent. The catalyst is reusable.

Studies on Metallocene Polyolefin and Polyvinyl Chloride for Blood and Blood Component Storage Applications

Plasticized poly(vinyl chloride) (pPVC), although a major player in the medical field, is at present facing lot of criticism due to some of its limitations like the leaching out of the toxic plasticizer, di ethylhexyl phthalate (DEHP) to the medium and the emission of an environmental pollutant,dioxin gas,at the time of the post use disposal of PVC Products by incineration. Due to these reasons, efforts are on to reduce the use of pPVC considerably in the medical field and to find viable alternative materials. The present study has been undertaken in this context to find a suitable material for the manufacture of medical aids in place of pPVC. The main focus of this study has been to find out a non-DEHP material as plasticizer for pPVC and another suitable material for the complete repalcement of pPVC for blood/ blood component storage applications.Two approaches have been undertaken for this purpose-(1)the controversial plasticizer, DEHP has been partially replaced by polymeric plasticizers(2) an alternative material, namely, metallocene polyolefin (mPO) has been used and suitably modified to match the properties of flexible PVC used for blood and blood component storage applications.

GABAA and GABAB Receptor Gene Expression and Functional Regulation During Pancreatic Regeneration and Insulin Secretion in Rats

The present study demonstrate the functional alterations of the GABAA and GABAB receptors and the gene expression during the regeneration of pancreas following partial pancreatectomy. The role of these receptors in insulin secretion and pancreatic DNA synthesis using the specific agonists and antagonists also are studied in vitro. The alterations of GABAA and GABAR receptor function and gene expression in the brain stem, crebellum and hypothalamus play an important role in the sympathetic regulation of insulin secretion during pancreatic regeneration. Previous studies have given much information linking functional interaction between GABA and the peripheral nervous system. The involvement of specific receptor subtypes functional regulation during pancreatic regeneration has not given emphasis and research in this area seems to be scarce. We have observed a decreased GABA content, down regulation of GABAA receptors and an up regulation of GABAB receptors in the cerebral cortex, brain stem and hypothalamus. Real Time-PCR analysis confirmed the receptor data in the brain regions. These alterations in the GABAA and GABAB receptors of the brain are suggested to govern the regenerative response and growth regulation of the pancreas through sympathetic innervation. In addition, receptor binding studies and Real Time-PCR analysis revealed that during pancreatic regeneration GABAA receptors were down regulated and GABAB receptors were up regulated in pancreatic islets. This suggests an inhibitory role for GABAA receptors in islet cell proliferation i.e., the down regulation of this receptor facilitates proliferation. Insulin secretion study during 1 hour showed GABA has inhibited the insulin secretion in a dose dependent manner in normal and hyperglycaemic conditions. Bicuculline did not antagonize this effect. GABAA agonist, muscimol inhibited glucose stimulated insulin secretion from pancreatic islets except in the lowest concentration of 1O-9M in presence of 4mM glucose.Musclmol enhanced insulin secretion at 10-7 and 10-4M muscimol in presence of 20mM glucose- 4mM glucose represents normal and 20mM represent hyperglycaemic conditions. GABAB agonist, baclofen also inhibited glucose induced insulin secretion and enhanced at the concentration of 1O-5M at 4mM glucose and at 10-9M baclofen in presence of 20mM glucose. This shows a differential control of the GABAA and GABAB receptors over insulin release from the pancreatic islets. During 24 hours in vitro insulin secretion study it showed that low concentration of GABA has inhibited glucose stimulated insulin secretion from pancreatic islets. Muscimol, the GABAA agonist, inhibited the insulin secretion but, gave an enhanced secretion of insulin in presence of 4mM glucose at 10-7 , 10-5 and 1O-4M muscimol. But in presence of 20mM glucose muscimol significantly inhibited the insulin secretion. GABAB agonist, baclofen also inhibited glucose induced insulin secretion in presence of both 4mM and 20mM glucose. This shows the inhibitory role of GABA and its specific receptor subtypes over insulin synthesis from pancreatic bete-islets. In vitro DNA synthesis studies showed that activation of GABAA receptor by adding muscimol, a specific agonist, inhibited islet DNA synthesis. Also, the addition of baclofen, a specific agonist of GABAB receptor resulted in the stimulation of DNA synthesis.Thus the brain and pancreatic GABAA and GABAB receptor gene expression differentially regulates pancreatic insulin secretion and islet cell proliferation during pancreatic regeneration. This will have immense clinical significance in therapeutic applications in the management of Diabetes mellitus.

Investigations on nanosecond laser produced plasma in air from the multi-component material YBa2Cu3O7

The laser produced plasma from the multi-component target YBa2CU3O7 was analyzed using Michelson interferometry and time resolved emission spectroscopy. The interaction of 10 ns pulses of 1.06 mum radiation from a Q-switched Nd:YAG laser at laser power densities ranging from 0.55 GW cm-2 to 1.5 GW cm-2 has been studied. Time resolved spectral measurements of the plasma evolution show distinct features at different points in its temporal history. For a time duration of less than 55 ns after the laser pulse (for a typical laser power density of 0.8 GW cm-2, the emission spectrum is dominated by black-body radiation. During cooling after 55 ns the spectral emission consists mainly of neutral and ionic species. Line averaged electron densities were deduced from interferometric line intensity measurements at various laser power densities. Plasma electron densities are of the order of 1017 cm-3 and the plasma temperature at the core region is about 1 eV. The measurement of plasma emission line intensities of various ions inside the plasma gave evidence of multiphoton ionization of the elements constituting the target at low laser power densities. At higher laser power densities the ionization mechanism is collision dominated. For elements such as nitrogen present outside the target, ionization is due to collisions only.

Dopamine D2 and 5-HT2A receptor gene expression

Neuronal dopamine and serotonin receptors are widely distributed in the central and the peripheral nervous systems at different levels. Dopaminergic and serotonergic systems have crucial role in aldehyde dehydrogenase regulation Stimulation of autonomic nervous system during ethanol treatment is suggested to be an important factor in regulating the ALDH function. The ALDH enzyme activity was increased in plasma, cerebral cortex, and liver but decreased in cerebellum. The ALDH enzyme affinity was decreased in plasma, brainstem and liver and increased in cerebral cortex and cerebellum. Dopamine and serotonin content decreased in liver and brain regions - cerebral cortex, corpus striatum of ethanol treated rats with an increased HVA/DA, 5-HIAA/5-HT tumover rate. Dopamine content decreased in brainstem with an increased HVA/DA turnover rate and serotonin content decreased with an increased 5-HIAA/5-HT turnover rate in the brainstem of ethanol treated rats compared to control. Serotonin content increased in hypothalamus with a decreased 5-HIAA/5—HT turnover rate where as dopamine content decreased in hypothalamus with an increased HVA/DA tumover rate of ethanol treated rats compared to control.alterations of DA D2 and 5-HTQA receptor function and gene expression in the cerebellum, hypothalamus, corpus striatum, cerebral cortex play an important role in the sympathetic regulation of ALDH enzyme in ethanol addiction. There is a serotonergic and dopaminergic functional regulation of ALDH activity in the brain regions and liver of ethanol treated rats. Gene expression studies of DA D2 and 5'HT2A studies confirm these observations. Perfusion studies using DA, 5-HT and glucose showed ALDH regulatory function. Brain activity measeurement using EEG showed a prominentfrontal brain wave difference. This will have immense clinical significance in the management of ethanol addiction.

An Efficient Multi-component Synthesis of 6-Amino-3-methyl-4-Aryl-2,4- dihydropyrano[2,3-c]Pyrazole-5-carbonitriles

Multi-component reactions are effective in building complex molecules in a single step in a minimum amount of time and with facile isolation procedures; they have high economy1–7 and thus have become a powerful synthetic strategy in recent years.8–10 The multicomponent protocols are even more attractive when carried out in aqueous medium. Water offers several benefits, including control over exothermicity, and the isolation of products can be carried out by single phase separation technique. Pyranopyrazoles are a biologically important class of heterocyclic compounds and in particular dihydropyrano[2,3-c]pyrazoles play an essential role in promoting biological activity and represent an interesting template in medicinal chemistry. Heterocyclic compounds bearing the 4-H pyran unit have received much attention in recent years as they constitute important precursors for promising drugs.11–13 Pyrano[2,3-c]pyrazoles exhibit analgesic,14 anti-cancer,15 anti-microbial and anti-inflammatory16 activity. Furthermore dihydropyrano[2,3-c]pyrazoles show molluscidal activity17,18 and are used in a screening kit for Chk 1 kinase inhibitor activity.19,20 They also find applications as pharmaceutical ingredients and bio-degradable agrochemicals.21–29 Junek and Aigner30 first reported the synthesis of pyrano[2,3-c]pyrazole derivatives from 3-methyl-1-phenylpyrazolin-5-one and tetracyanoethylene in the presence of triethylamine. Subsequently, a number of synthetic approaches such as the use of triethylamine,31 piperazine,32 piperidine,33 N-methylmorpholine in ethanol,34 microwave irradiation,35,36 solvent-free conditions,37–39 cyclodextrins (CDs),40 different bases in water,41 γ -alumina,42 and l-proline43 have been reported for the synthesis of 6-amino-4-alkyl/aryl-3-methyl- 2,4-dihydropyrano[2,3-c]pyrazole-5-carbonitriles. Recently, tetraethylammonium bromide (TEABr) has emerged as mild, water-tolerant, eco-friendly and inexpensive catalyst. To the best of our knowledge, quaternary ammonium salts, more specifically TEABr, have notbeen used as catalysts for the synthesis of pyrano[2,3-c]pyrazoles, and we decided to investigate the application of TEABr as a catalyst for the synthesis of a series of pyrazole-fused pyran derivatives via multi-component reactions

Spectral clustering independent component analysis for tissue classification from brain MRI

A spectral angle based feature extraction method, Spectral Clustering Independent Component Analysis (SC-ICA), is proposed in this work to improve the brain tissue classification from Magnetic Resonance Images (MRI). SC-ICA provides equal priority to global and local features; thereby it tries to resolve the inefficiency of conventional approaches in abnormal tissue extraction. First, input multispectral MRI is divided into different clusters by a spectral distance based clustering. Then, Independent Component Analysis (ICA) is applied on the clustered data, in conjunction with Support Vector Machines (SVM) for brain tissue analysis. Normal and abnormal datasets, consisting of real and synthetic T1-weighted, T2-weighted and proton density/fluid-attenuated inversion recovery images, were used to evaluate the performance of the new method. Comparative analysis with ICA based SVM and other conventional classifiers established the stability and efficiency of SC-ICA based classification, especially in reproduction of small abnormalities. Clinical abnormal case analysis demonstrated it through the highest Tanimoto Index/accuracy values, 0.75/98.8%, observed against ICA based SVM results, 0.17/96.1%, for reproduced lesions. Experimental results recommend the proposed method as a promising approach in clinical and pathological studies of brain diseases

Brain Tissue Classification from Multispectral MRI by Wavelet based Principal Component Analysis

In this paper, we propose a multispectral analysis system using wavelet based Principal Component Analysis (PCA), to improve the brain tissue classification from MRI images. Global transforms like PCA often neglects significant small abnormality details, while dealing with a massive amount of multispectral data. In order to resolve this issue, input dataset is expanded by detail coefficients from multisignal wavelet analysis. Then, PCA is applied on the new dataset to perform feature analysis. Finally, an unsupervised classification with Fuzzy C-Means clustering algorithm is used to measure the improvement in reproducibility and accuracy of the results. A detailed comparative analysis of classified tissues with those from conventional PCA is also carried out. Proposed method yielded good improvement in classification of small abnormalities with high sensitivity/accuracy values, 98.9/98.3, for clinical analysis. Experimental results from synthetic and clinical data recommend the new method as a promising approach in brain tissue analysis.

Wavelet based Independent Component Analysis for Multispectral Brain Tissue Classification

Multispectral analysis is a promising approach in tissue classification and abnormality detection from Magnetic Resonance (MR) images. But instability in accuracy and reproducibility of the classification results from conventional techniques keeps it far from clinical applications. Recent studies proposed Independent Component Analysis (ICA) as an effective method for source signals separation from multispectral MR data. However, it often fails to extract the local features like small abnormalities, especially from dependent real data. A multisignal wavelet analysis prior to ICA is proposed in this work to resolve these issues. Best de-correlated detail coefficients are combined with input images to give better classification results. Performance improvement of the proposed method over conventional ICA is effectively demonstrated by segmentation and classification using k-means clustering. Experimental results from synthetic and real data strongly confirm the positive effect of the new method with an improved Tanimoto index/Sensitivity values, 0.884/93.605, for reproduced small white matter lesions

Identifying the component structure of job satisfaction by principal components analysis among extension officers in North West Province, South Africa

The component structure of a 34-item scale measuring different aspects of job satisfaction was investigated among extension officers in North West Province, South Africa. A simple random sampling technique was used to select 40 extension officers from which data were collected. A structured questionnaire consisting of 34 job satisfaction and 10 personal characteristic items was administered to the extension officers. Items on job satisfaction were measured at interval level and analyzedwith Principal ComponentAnalysis. Most of the respondents (82.5%) weremales, between 40 to 45 years, 85% were married and 87.5% had a diploma as their educational qualification. Furthermore, 54% of the households size between 4 to 6 persons, whereas 75% were Christians. The majority of the extension officers lived in their job area (82.5), while 80% covered at least 3 communities and 3 farmer groups. In terms of number of farmers covered, only 40% of the extension officers covered more than 500 farmers and 45% travelled more than 40 km to reach their farmers. From the job satisfaction items 9 components were extracted to show areas for job satisfaction among extension officers. These were in-service training, research policies, communicating recommended practices, financial support for self and family, quality of technical help, opportunity to advance education, management and control of operations, rewarding system and sanctions. The results have several implications for motivating extension officers for high job performance especially with large number of clients and small number of extension agents.

Analysis of second messengers in peripheral and central circadian pacemakers

In früheren Experimenten konnte gezeigt werden, dass die sekundären Botenstoffe cAMP, cGMP und IP3 in der olfaktorischen Signaltransduktionskaskade bei Manduca sexta involviert sind. Während cGMP Perfusionen in die Antenne die Pheromonwahrnehmung tageszeitabhängig adaptierten, führten cAMP Perfusionen zu einer tageszeitabhängigen Sensitisierung, ähnlicher der von Octopamin (OA). Daher wurde hypothetisiert, dass eine tageszeitabhängige Oszillation antennaler OA Level sowie der intrazelluläre Kalziumkonzentration in einer Schwankung von sekundären Botenstoffen resultieren könnte. Diese Hypothese wurde mittels biochemischen Nachweißverfahren in der Antenne von M. sexta und Rhyparobia maderae überprüft. Tatsächlich konnten in der Antenne des Tabakschwärmers tageszeitabhängige Unterschiede in der OA-, cAMP- und IP3-, aber nicht in der cGMP Konzentration, nachgewiesen werden. Während die cAMP- und OA Oszillationen einander sehr ähnelten und die Maxima in der Paarungsphase aufzeigten, korrelierte der IP3 Verlauf sehr stark mit dem Flug- bzw. Fressverhalten. Diese Korrelationen konnten auch in der Madeira Schabe beobachtet werden, in der darüber hinaus gezeigt werden konnte, dass antennale cAMP- und IP3 Level von dem circadianen Uhrwerk gesteuert werden. Zudem wurde herausgefunden, dass OA die cAMP- und teilweise auch die IP3- Spiegel reguliert. Demgegenüber beeinflusste Kalzium die Konzentration aller untersuchten sekundären Botenstoffe. Daher wird angenommen, dass die intrazelluläre Kalziumkonzentration aber auch der antennale OA Level kritische Faktoren bei der Regelung der olfaktorischen Sensitivität sind. Da Oszillationen von sekundären Botenstoffen in mutmaßlichen, peripheren Schrittmacher nachgewiesen wurden, wurde untersucht, ob sie auch im circadianen Schrittmacher der Madeira Schabe oszillieren und ob das Neuropeptid pigment-dispersing factor (PDF), ein entscheidender Kopplungsfaktor des Uhrwerks in Insekten, diese Rhythmen generieren könnte. Es konnte gezeigt werden, dass PDF die cAMP Synthese steigert. Darüber hinaus wurden bimodale cAMP Oszillationen unter licht-dunkel Bedingungen beobachtet, welche unter konstanten Umweltbedingungen verblieben. Daher wird angenommen, dass PDF Freisetzung zelluläres cAMP erhöht über das das circadiane Uhrwerk synchronisiert wird.

Physiological analysis of central and peripheral insect circadian pacemaker neurons

Alle bisher untersuchten Lebewesen besitzen (circadiane) innere Uhren, die eine endogene Perioden-länge von ungefähr 24 Stunden generieren. Eine innere Uhr kann über Zeitgeber mit der Umwelt synchronisiert werden und ermöglicht dem Organismus, rhythmische Umweltveränderungen vorweg zu nehmen. Neben einem zentralen Schrittmacher, der Physiologie und Verhalten des Organismus steuert, gibt es in unterschiedlichen Organen auch periphere Uhren, die die zeitlichen Abläufe in der spezifischen Funktion dieser Organe steuern. In dieser Arbeit sollten zentrale und periphere Schrittmacherneurone von Insekten physiologisch untersucht und verglichen werden. Die Neurone der akzessorischen Medulla (AME) von Rhyparobia maderae dienten als Modellsystem für zentrale Schrittmacher, während olfaktorische Rezeptorneurone (ORNs) von Manduca sexta als Modellsystem für periphere Schrittmacher dienten. Die zentralen Schrittmacherneurone wurden in extrazellulären Ableitungen an der isolierten AME (Netzwerkebene) und in Patch-Clamp Experimenten an primären AME Zellkulturen (Einzelzellebene) untersucht. Auf Netzwerkebene zeigten sich zwei charakteristische Aktivitätsmuster: regelmäßige Aktivität und Wechsel zwischen hoher und niedriger Aktivität (Oszillationen). Es wurde gezeigt, dass Glutamat ein Neurotransmitter der weitverbreiteten inhibitorischen Synapsen der AME ist, und dass in geringem Maße auch exzitatorische Synapsen vorkommen. Das Neuropeptid pigment-dispersing factor (PDF), das von nur wenigen AME Neuronen exprimiert wird und ein wichtiger Kopplungsfaktor im circadianen System ist, führte zu Hemmungen, Aktivierungen oder Oszillationen. Die Effekte waren transient oder langanhaltend und wurden wahrscheinlich durch den sekundären Botenstoff cAMP vermittelt. Ein Zielmolekül von cAMP war vermutlich exchange protein directly activated by cAMP (EPAC). Auf Einzelzellebene wurde gezeigt, dass die meisten AME Neurone depolarisiert waren und deshalb nicht feuerten. Die Analyse von Strom-Spannungs-Kennlinien und pharmakologische Experimente ergaben, dass unterschiedliche Ionenkanäle vorhanden waren (Ca2+, Cl-, K+, Na+ Kanäle sowie nicht-spezifische Kationenkanäle). Starke, bei hohen Spannungen aktivierende Ca2+ Ströme (ICa) könnten eine wichtige Rolle bei Ca2+-abhängiger Neurotransmitter-Ausschüttung, Oszillationen, und Aktionspotentialen spielen. PDF hemmte unterschiedliche Ströme (ICa, IK und INa) und aktivierte nicht-spezifische Kationenströme (Ih). Es wurde angenommen, dass simultane PDF-abhängige Hyper- und Depolarisationen rhythmische Membranpotential-Oszillationen verursachen. Dieser Mechanismus könnte eine Rolle bei PDF-abhängigen Synchronisationen spielen. Die Analyse peripherer Schrittmacherneurone konzentrierte sich auf die Charakterisierung des olfaktorischen Corezeptors von M. sexta (MsexORCO). In anderen Insekten ist ORCO für die Membran-Insertion von olfaktorischen Rezeptoren (ORs) erforderlich. ORCO bildet Komplexe mit den ORs, die in heterologen Expressionssystemen als Ionenkanäle fungieren und Duft-Antworten vermitteln. Es wurde die Hypothese aufgestellt, dass MsexORCO in pheromonsensitiven ORNs in vivo nicht als Teil eines ionotropen Rezeptors sondern als Schrittmacherkanal fungiert, der unterschwellige Membranpotential-Oszillationen generiert. MsexORCO wurde mit vermeintlichen Pheromonrezeptoren in human embryonic kidney (HEK 293) Zellen coexprimiert. Immuncytochemie und Ca2+ Imaging Experimente zeigten sehr schwache Expressionsraten. Trotzdem war es möglich zu zeigen, dass MsexORCO wahrscheinlich ein spontan-aktiver, Ca2+-permeabler Ionenkanal ist, der durch den ORCO-Agonisten VUAA1 und cyclische Nucleotide aktiviert wird. Außerdem wiesen die Experimente darauf hin, dass MsexOR-1 offensichtlich der Bombykal-Rezeptor ist. Eine weitere Charakterisierung von MsexORCO in primären M. sexta ORN Zellkulturen konnte nicht vollendet werden, weil die ORNs nicht signifikant auf ORCO-Agonisten oder -Antagonisten reagierten.

Component based recognition of objects in an office environment

We present a component-based approach for recognizing objects under large pose changes. From a set of training images of a given object we extract a large number of components which are clustered based on the similarity of their image features and their locations within the object image. The cluster centers build an initial set of component templates from which we select a subset for the final recognizer. In experiments we evaluate different sizes and types of components and three standard techniques for component selection. The component classifiers are finally compared to global classifiers on a database of four objects.