Assessment of the breakdown products of solar/UV induced photolytic degradation of food dye tartrazine

The food dye tartrazine (CI 19140) was exposed to UV irradiation from an artificial source, a mercury vapor lamp, and a natural one, sunlight. It was observed that conditions such as energy dose, irradiation time, pH and initial dye concentration affected its discoloration. There was 100% of color removal, after 30 min of irradiation, when a dye solution 1 x 10(-5) mol L-1 was submitted to an energy dose of 37.8 J cm(-2). Liquid Chromatography coupled to Diode Array Detection and Mass Spectrometry confirmed the cleavage of the chromophore group and the formation of five by-products at low concentration. Although by-products were formed, the Salmonella/microsome mutagenicity assay performed for both, the dye solution at a dose of 5.34 mg/plate and the solutions obtained after exposure to UV irradiation, did not present mutagenic activity for TA98 and TA100 with and without S9. (C) 2014 Elsevier Ltd. All rights reserved.

Protonation Pattern, Tautomerism, Conformerism, and Physicochemical Analysis in New Crystal Forms of the Antibiotic Doxycycline

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

SEQUENTIAL SAMPLING AND MICROSATELLITES FOR EVALUATION OF THE GENETIC QUALITY IN CORN SEEDS LOTS

In the seed production system, genetic purity is one of the fundamental requirements for its commercialization. The present work had the goal of determined the sample size for genetic purity evaluation, in order to protect the seed consumer and the producer and to evaluate the sensitivity of microsatellite technique for discriminating hybrids from their respective relatives and for detecting mixtures when they are present in small amounts in the samples. For the sequential sampling, hybrid seeds were marked and mixed in with the seed lots, simulating the following levels of contamination: 0.25, 0.5, 1.0, 2.0, 4.0, and 6.0%. After this, groups of 40 seeds were taken in sequence, up to a maximum of 400 seeds, with the objective of determining the quantity of seeds necessary to detect the percentage of mixture mentioned above. The sensitivity of microsatellite technique was evaluated by mixing different proportions of DNA from the hybrids with their respective seed lines. For the level of mixture was higher than 1:8 (1P1:8P2; 8P1:1P2), the sensitivity of the marker in detecting different proportions of the mixture varied according to the primer used. In terms of the sequential sampling, it was verified that in order to detect mixture levels higher than 1% within the seed lot- with a risk level for both the producer and the consumer of 0.05- the size of the necessary sample was smaller than the size needed for the fixed sample size. This also made it possible to reduce costs, making it possible to use microsatellites to certify the genetic purity of corn seeds lots.

Refinement and partial validation of the UNESP-Botucatu multidimensional composite pain scale for assessing postoperative pain in horses

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effect of the consumption of a symbiotic shake on the intestinal microflora of erdely people

The consumption of foods containing probiotic and prebiotic ingredients is growing consistently every year, and in view of the limited number of studies investigating their effect in the elderly. Objective The objective of this study was to evaluate the effect of the consumption of a symbiotic shake containing Lactobacillus acidophilus, Bifidobacterium bifidum and fructooligosaccharides on glycemia and cholesterol levels in elderly people. Methods A randomized, double-blind, placebo-controlled study was conducted on twenty volunteers (ten for placebo group and ten for symbiotic group), aged 50 to 60 years. The criteria for inclusion in the study were: total cholesterol > 200 mg/dL; triglycerides > 200 mg/dL and glycemia > 110 mg/dL. Over a total test period of 30 days, 10 individuals (the symbiotic group) consumed a daily dose of 200 mL of a symbiotic shake containing 108 UFC/mL Lactobacillus acidophilus, 108 UFC/mL Bifidobacterium bifidum and 2 g oligofructose, while 10 other volunteers (the placebo group) drank daily the same amount of a shake that did not contain any symbiotic bacteria. Blood samples were collected 15 days prior to the start of the experiment and at 10-day intervals after the beginning of the shake intake. The standard lipid profile (total cholesterol, triglycerides and HDL cholesterol) and glycemia, or blood sugar levels, were evaluated by an enzyme colorimetric assay. Results The results of the symbiotic group showed a non-significant reduction (P > 0.05) in total cholesterol and triglycerides, a significant increase (P < 0.05) in HDL cholesterol and a significant reduction (P < 0.05) in fasting glycemia. No significant changes were observed in the placebo group. Conclusion The consumption of symbiotic shake resulted in a significant increase in HDL and a significant decrease of glycemia.

Regulation of the abundance and turnover of copepod species by temperature, turbidity and habitat type in a large river basin

Freshwater copepods were sampled in the La Plata River basin to identify the processes that affect beta diversity and to determine the main factors influencing their geographical distribution and patterns of endemism. Beta diversity patterns exhibited strong dissimilarity between locations; the turnover process was predominant and indicated a replacement of species along the basin. Redundancy analysis indicated the presence of two large sets of species separated geographically by a boundary zone, with several associated variables. Northern species were associated with water transparency and temperature, mean air temperature, mean air temperature during winter and minimum air temperature of coldest month, indicating that these species are not tolerant to low temperatures and are abundant in reservoirs that are common in the upper stretch of the Paraná River basin. Southern species were related with amplitude of air temperature, turbidity, total phosphorus and total suspended matter, indicating that these species are polythermic and have adapted to live in river stretches. From 20 environmental variables analyzed in our study, partial least squares analysis indicated four variables with increased retention of effects on copepod abundance: air temperature, minimum temperature of coldest month, turbidity and transparency. Because almost all of the species found in this study occurred across a wide range of habitat types, the cause of the separation between river and reservoir species could be considered to be more anthropogenic than natural, and it primarily affected species abundance. For certain members of the northern group of copepod species, distribution was dependent on high temperatures, whereas the distribution of the southern group indicated that the species were polythermic.

The Specific Status of the Narrow-Skulled Vole (Subgenus Stenocranius Kashchenko) in North America

On morphological and zoogeographical grounds, discussed in the present paper, it is concluded that the narrow-skulled vole in North America, previously designated Microtus (Stenocranius) miurus Osgood, is conspecific with the Eurasian M. (Stenocranius) gregalis Pallas. Fourteen subspecies in Eurasia and 5 in North America are now recognized, but it is probable that the number in Eurasia will be reduced through future investigation. The Eurasian subspecies of this vole comprise two major groups, of which one occupies the tundra zone and the other occurs across central Asia below latitude 60° N; their geographic ranges are largely separate but evidently become confluent in northeastern Siberia. The members of the northern group of Eurasian subspecies and the North American forms are closely related; the present distribution of the latter indicates post-glacial dispersal from the Amphiberingian Refugium. It is believed that the tundra-inhabiting voles in Eurasia likewise survived the Pleistocene glaciations in northern refugia, while the members of the southern group of subspecies probably represent populations that survived south of the limits of the continental glaciers. The ranges of the two Eurasian groups probably have become confluent during post-glacial time in northeastern Siberia as a result of the southward spread of the northern forms. At least, the subspecies having the intervening range closely resembles members of the northern group. Some of the ecological and ethological characteristics of these voles are briefly discussed. The chromosome number of one of the North American subspecies of narrow-skulled vole was determined to be 54; this is the first time that the chromosomes of a member of the subgenus Stenocranius have been investigated. A karyogram has been included. German abstract: Auf morphologischen und tiergeographischen Grundlagen, die in dieser Arbeit besprochen wurden, ist festgestellt worden, daß die schmalschädlige Wiihlmaus in Nordamerika, friiher Microtus (Stenocranius) miurus Osgood bezeichnet, mit der palaearktischen Art M. (Stenocranius) gregalis Pallas identisch ist. Zur Zeit gelten 14 Unterarten in Eurasien und 5 in Nordamerika als unterscheidbar; vermutlich aber wird die Zahl der palaearktischen Unterarten durch eingehendere Untersuchungen künftig vermindert werden. Auf Grund ihrer Verbreitung bilden die palaearktischen Unterarten zwei beinahe vollständig getrennte Gruppen. Die Wühlmäuse der nördlichen Gruppe bewohnen die Tundrazone, während die Vertreter der zweiten Gruppe über Mittelasien südlicher als 60° N.B. verbreitet sind. Die Verbreitungsgebiete der zwei Gruppen verbinden sich anscheinend. Die nordamerikanischen schmalschädligen Wühlmäuse sind mit den in der Tundrazone vorkommenden palaearktischen Formen nahe verwandt; sie haben sich wahrscheinlich während der Postglazialzeit aus dem Amphiberingschen Refugium verbreitet. Möglicherweise überlebten die tundrabewohnenden Wühlmäuse Eurasiens die Eiszeit ebenfalls in vereinzelten Refugien in Nordostsibirien, während die Formen der südlichen Gruppe sie jenseits der Grenzen des Festlandsgletschers überlebten. Wahrscheinlich wurden die zwei Verbreitungsgebiete dieser Art in Eurasien erst während der Postglazialzeit durch das Vordringen der nordischen Formen verbunden, da eine nähere Verwandtschaft zwischen den nördlichen und der dazwischenliegenden Unterart besteht. Einige ökologische und ethologische Eigentümlichkeiten dieser Wühlmäuse werden kurz besprochen. Es wurde festgestellt, daß eine der nordamerikanischen Unterarten der schmalschädligen Wühlmaus 54 Chromosomen hat; sie ist der einzige Vertreter der Untergattung Stenocranius, dessen Chromosomen untersucht worden sind.

Analysis of the mRNA expression of the S100 beta protein in adipocytes of patients with diabetes mellitus, type 2

Objective: This study aims to explore the possible relationship between the expression level of S100 beta protein mRNA with diabetes mellitus type 2 in adipocytes from patients with this disease in comparison with normoglycemic individuals. Materials and methods: Samples of adipose tissue of eight patients from the coronary section of the Institute Dante Pazzanese of Cardiology (IDPC), four in Group Diabetes and four of Normoglycemic group, were evaluated by RT-PCR real time. Results: An increase around 15 times values, between the threshold cycle (Delta Ct), of mRNA expression of S100 beta protein in adipocytes of the diabetes group was observed in comparison to the control group (p = 0.015). Conclusion: Our results indicate, for the first time, that there is coexistence of increased expression of the S100 beta and the type 2 diabetes mellitus gene. Arq Bras Endocrinol Metab. 2012;56(7):435-40

X-ray diffraction and theoretical investigation of the Gedunin crystal structure

The Gedunin compound (C28H34O6) is a natural product extracted from Trichilia pallida that has shown a wide activity. The crystallographic structure shows two conformers in the asymmetric unit, which differ in a rotation of the furan group. To understand this molecular arrangement, the density functional calculations. Molecular Electrostatic Potential (MEP) and thermodynamic function calculation have been performed at the B3LYP/6-311++g(d,p) level. Both conformers were optimized and the agreement with the experimental structure was very good, making possible further theoretical analysis of the structure. The inter-conversion between two conformers depends on the energy barrier. This process is studied in the vacuum and shows two transition states with a low energetic barrier for a potential energy curve scanning rigid around furan group: 4.37 kcal/mol and 16.52 kcal/mol. As the first transition state has a notably lower energetic barrier, the preferred inter-conversion pathway between the conformers involves the first rather than the second transition state. Understanding this transition state in detail led us to perform its optimization, showing an energetic barrier around 3.66 kcal/mol. The negative free energy and low enthalpy confirm that the process is spontaneous and exothermic. The results show that this requirement makes the existence of the two conformers in the asymmetric unit possible. The structure of molecules in the asymmetric unit is better understood when the MEP is used on the interaction between molecules. For Gedunin, both molecules have shown MEP with well-defined regions, and this behavior contributes to the observed link between molecules and for the negative regions complementing positive regions of another molecule. (C) 2011 Elsevier B.V. All rights reserved.

Stellar Relics of the hierarchical assembly of the Galaxy

This PhD Thesis is part of a long-term wide research project, carried out by the "Osservatorio Astronomico di Bologna (INAF-OABO)", that has as primary goal the comprehension and reconstruction of formation mechanism of galaxies and their evolution history. There is now substantial evidence, both from theoretical and observational point of view, in favor of the hypothesis that the halo of our Galaxy has been at least partially, built up by the progressive accretion of small fragments, similar in nature to the present day dwarf galaxies of the Local Group. In this context, the photometric and spectroscopic study of systems which populate the halo of our Galaxy (i.e. dwarf spheroidal galaxy, tidal streams, massive globular cluster, etc) permits to discover, not only the origin and behaviour of these systems, but also the structure of our Galactic halo, combined with its formation history. In fact, the study of the population of these objects and also of their chemical compositions, age, metallicities and velocity dispersion, permit us not only an improvement in the understanding of the mechanisms that govern the Galactic formation, but also a valid indirect test for cosmological model itself. Specifically, in this Thesis we provided a complete characterization of the tidal Stream of the Sagittarius dwarf spheroidal galaxy, that is the most striking example of the process of tidal disruption and accretion of a dwarf satellite in to our Galaxy. Using Red Clump stars, extracted from the catalogue of the Sloan Digital Sky Survey (SDSS) we obtained an estimate of the distance, the depth along the line of sight and of the number density for each detected portion of the Stream (and more in general for each detected structure along our line of sight). Moreover comparing the relative number (i.e. the ratio) of Blue Horizontal Branch stars and Red Clump stars (the two features are tracers of different age/different metallicity populations) in the main body of the galaxy and in the Stream, in order to verify the presence of an age-metallicity gradient along the Stream. We also report the detection of a population of Red Clump stars probably associated with the recently discovered Bootes III stellar system. Finally, we also present the results of a survey of radial velocities over a wide region, extending from r ~ 10' out to r ~ 80' within the massive star cluster Omega Centauri. The survey was performed with FLAMES@VLT, to study the velocity dispersion profile in the outer regions of this stellar system. All the results presented in this Thesis, have already been published in refeered journals.

Biochemical analysis of the phosphatase domain of the human soluble epoxide hydrolase (sEH)

Die lösliche Epoxidhydrolase (sEH) gehört zur Familie der Epoxidhydrolase-Enzyme. Die Rolle der sEH besteht klassischerweise in der Detoxifikation, durch Umwandlung potenziell schädlicher Epoxide in deren unschädliche Diol-Form. Hauptsächlich setzt die sEH endogene, der Arachidonsäure verwandte Signalmoleküle, wie beispielsweise die Epoxyeicosatrienoic acid, zu den entsprechenden Diolen um. Daher könnte die sEH als ein Zielenzym in der Therapie von Bluthochdruck und Entzündungen sowie diverser anderer Erkrankungen eingesetzt werden. rnDie sEH ist ein Homodimer, in dem jede Untereinheit aus zwei Domänen aufgebaut ist. Das katalytische Zentrum der Epoxidhydrolaseaktivität befindet sich in der 35 kD großen C-terminalen Domäne. Dieser Bereich der sEH s wurde bereits im Detail untersucht und nahezu alle katalytischen Eigenschaften des Enzyms sowie deren dazugehörige Funktionen sind in Zusammenhang mit dieser Domäne bekannt. Im Gegensatz dazu ist über die 25 kD große N-terminale Domäne wenig bekannt. Die N-terminale Domäne der sEH wird zur Haloacid Dehalogenase (HAD) Superfamilie von Hydrolasen gezählt, jedoch war die Funktion dieses N-terminal Domäne lange ungeklärt. Wir haben in unserer Arbeitsgruppe zum ersten Mal zeigen können, dass die sEH in Säugern ein bifunktionelles Enzym ist, welches zusätzlich zur allgemein bekannten Enzymaktivität im C-terminalen Bereich eine weitere enzymatische Funktion mit Mg2+-abhängiger Phosphataseaktivität in der N-terminalen Domäne aufweist. Aufgrund der Homologie der N-terminalen Domäne mit anderen Enzymen der HAD Familie wird für die Ausübung der Phosphatasefunktion (Dephosphorylierung) eine Reaktion in zwei Schritten angenommen.rnUm den katalytischen Mechanismus der Dephosphorylierung weiter aufzuklären, wurden biochemische Analysen der humanen sEH Phosphatase durch Generierung von Mutationen im aktiven Zentrum mittels ortsspezifischer Mutagenese durchgeführt. Hiermit sollten die an der katalytischen Aktivität beteiligten Aminosäurereste im aktiven Zentrum identifiziert und deren Rolle bei der Dephosphorylierung spezifiziert werden. rnrnAuf Basis der strukturellen und möglichen funktionellen Ähnlichkeiten der sEH und anderen Mitgliedern der HAD Superfamilie wurden Aminosäuren (konservierte und teilweise konservierte Aminosäuren) im aktiven Zentrum der sEH Phosphatase-Domäne als Kandidaten ausgewählt.rnVon den Phosphatase-Domäne bildenden Aminosäuren wurden acht ausgewählt (Asp9 (D9), Asp11 (D11), Thr123 (T123), Asn124 (N124), Lys160 (K160), Asp184 (D184), Asp185 (D185), Asn189 (N189)), die mittels ortsspezifischer Mutagenese durch nicht funktionelle Aminosäuren ausgetauscht werden sollten. Dazu wurde jede der ausgewählten Aminosäuren durch mindestens zwei alternative Aminosäuren ersetzt: entweder durch Alanin oder durch eine Aminosäure ähnlich der im Wildtyp-Enzym. Insgesamt wurden 18 verschiedene rekombinante Klone generiert, die für eine mutante sEH Phosphatase Domäne kodieren, in dem lediglich eine Aminosäure gegenüber dem Wildtyp-Enzym ersetzt wurde. Die 18 Mutanten sowie das Wildtyp (Sequenz der N-terminalen Domäne ohne Mutation) wurden in einem Expressionsvektor in E.coli kloniert und die Nukleotidsequenz durch Restriktionsverdau sowie Sequenzierung bestätigt. Die so generierte N-terminale Domäne der sEH (25kD Untereinheit) wurde dann mittels Metallaffinitätschromatographie erfolgreich aufgereinigt und auf Phosphataseaktivität gegenüber des allgemeinen Substrats 4-Nitophenylphosphat getestet. Diejenigen Mutanten, die Phosphataseaktivität zeigten, wurden anschließend kinetischen Tests unterzogen. Basiered auf den Ergebnissen dieser Untersuchungen wurden kinetische Parameter mittels vier gut etablierter Methoden berechnet und die Ergebnisse mit der „direct linear blot“ Methode interpretiert. rnDie Ergebnisse zeigten, dass die meisten der 18 generierten Mutanten inaktiv waren oder einen Großteil der Enzymaktivität (Vmax) gegenüber dem Wildtyp verloren (WT: Vmax=77.34 nmol-1 mg-1 min). Dieser Verlust an Enzymaktivität ließ sich nicht durch einen Verlust an struktureller Integrität erklären, da der Wildtyp und die mutanten Proteine in der Chromatographie das gleiche Verhalten zeigten. Alle Aminosäureaustausche Asp9 (D9), Lys160 (K160), Asp184 (D184) und Asn189 (N189) führten zum kompletten Verlust der Phosphataseaktivität, was auf deren katalytische Funktion im N-terminalen Bereich der sEH hindeutet. Bei einem Teil der Aminosäureaustausche die für Asp11 (D11), Thr123 (T123), Asn124 (N124) und Asn185 (D185) durchgeführt wurden, kam es, verglichen mit dem Wildtyp, zu einer starken Reduktion der Phosphataseaktivität, die aber dennoch für die einzelnen Proteinmutanten in unterschiedlichem Ausmaß zu messen war (2 -10% and 40% of the WT enzyme activity). Zudem zeigten die Mutanten dieser Gruppe veränderte kinetische Eigenschaften (Vmax allein oder Vmax und Km). Dabei war die kinetische Analyse des Mutanten Asp11  Asn aufgrund der nur bei dieser Mutanten detektierbaren starken Vmax Reduktion (8.1 nmol-1 mg-1 min) und einer signifikanten Reduktion der Km (Asp11: Km=0.54 mM, WT: Km=1.3 mM), von besonderem Interesse und impliziert eine Rolle von Asp11 (D11) im zweiten Schritt der Hydrolyse des katalytischen Zyklus.rnZusammenfassend zeigen die Ergebnisse, dass alle in dieser Arbeit untersuchten Aminosäuren für die Phosphataseaktivität der sEH nötig sind und das aktive Zentrum der sEH Phosphatase im N-terminalen Bereich des Enzyms bilden. Weiterhin tragen diese Ergebnisse zur Aufklärung der potenziellen Rolle der untersuchten Aminosäuren bei und unterstützen die Hypothese, dass die Dephosphorylierungsreaktion in zwei Schritten abläuft. Somit ist ein kombinierter Reaktionsmechanismus, ähnlich denen anderer Enzyme der HAD Familie, für die Ausübung der Dephosphorylierungsfunktion denkbar. Diese Annahme wird gestützt durch die 3D-Struktur der N-terminalen Domäne, den Ergebnissen dieser Arbeit sowie Resultaten weiterer biochemischer Analysen. Der zweistufige Mechanismus der Dephosphorylierung beinhaltet einen nukleophilen Angriff des Substratphosphors durch das Nukleophil Asp9 (D9) des aktiven Zentrums unter Bildung eines Acylphosphat-Enzym-Zwischenprodukts, gefolgt von der anschließenden Freisetzung des dephosphorylierten Substrats. Im zweiten Schritt erfolgt die Hydrolyse des Enzym-Phosphat-Zwischenprodukts unterstützt durch Asp11 (D11), und die Freisetzung der Phosphatgruppe findet statt. Die anderen untersuchten Aminosäuren sind an der Bindung von Mg 2+ und/oder Substrat beteiligt. rnMit Hilfe dieser Arbeit konnte der katalytischen Mechanismus der sEH Phosphatase weiter aufgeklärt werden und wichtige noch zu untersuchende Fragestellungen, wie die physiologische Rolle der sEH Phosphatase, deren endogene physiologische Substrate und der genaue Funktionsmechanismus als bifunktionelles Enzym (die Kommunikation der zwei katalytischen Einheiten des Enzyms) wurden aufgezeigt und diskutiert.rn

Study of the helicity dependence of pi 0 photoproduction on proton at MAMI

The excitation spectrum is one of the fundamental properties of every spatially extended system. The excitations of the building blocks of normal matter, i.e., protons and neutrons (nucleons), play an important role in our understanding of the low energy regime of the strong interaction. Due to the large coupling, perturbative solutions of quantum chromodynamics (QCD) are not appropriate to calculate long-range phenomena of hadrons. For many years, constituent quark models were used to understand the excitation spectra. Recently, calculations in lattice QCD make first connections between excited nucleons and the fundamental field quanta (quarks and gluons). Due to their short lifetime and large decay width, excited nucleons appear as resonances in scattering processes like pion nucleon scattering or meson photoproduction. In order to disentangle individual resonances with definite spin and parity in experimental data, partial wave analyses are necessary. Unique solutions in these analyses can only be expected if sufficient empirical information about spin degrees of freedom is available. The measurement of spin observables in pion photoproduction is the focus of this thesis. The polarized electron beam of the Mainz Microtron (MAMI) was used to produce high-intensity, polarized photon beams with tagged energies up to 1.47 GeV. A "frozen-spin" Butanol target in combination with an almost 4π detector setup consisting of the Crystal Ball and the TAPS calorimeters allowed the precise determination of the helicity dependence of the γp → π0p reaction. In this thesis, as an improvement of the target setup, an internal polarizing solenoid has been constructed and tested. A magnetic field of 2.32 T and homogeneity of 1.22×10−3 in the target volume have been achieved. The helicity asymmetry E, i.e., the difference of events with total helicity 1/2 and 3/2 divided by the sum, was determined from data taken in the years 2013-14. The subtraction of background events arising from nucleons bound in Carbon and Oxygen was an important part of the analysis. The results for the asymmetry E are compared to existing data and predictions from various models. The results show a reasonable agreement to the models in the energy region of the ∆(1232)-resonance but large discrepancies are observed for energy above 600 MeV. The expansion of the present data in terms of Legendre polynomials, shows the sensitivity of the data to partial wave amplitudes up to F-waves. Additionally, a first, preliminary multipole analysis of the present data together with other results from the Crystal Ball experiment has been as been performed.

The venom composition of the parasitic wasp Chelonus inanitus resolved by combined expressed sequence tags analysis and proteomic approach

Background Parasitic wasps constitute one of the largest group of venomous animals. Although some physiological effects of their venoms are well documented, relatively little is known at the molecular level on the protein composition of these secretions. To identify the majority of the venom proteins of the endoparasitoid wasp Chelonus inanitus (Hymenoptera: Braconidae), we have randomly sequenced 2111 expressed sequence tags (ESTs) from a cDNA library of venom gland. In parallel, proteins from pure venom were separated by gel electrophoresis and individually submitted to a nano-LC-MS/MS analysis allowing comparison of peptides and ESTs sequences. Results About 60% of sequenced ESTs encoded proteins whose presence in venom was attested by mass spectrometry. Most of the remaining ESTs corresponded to gene products likely involved in the transcriptional and translational machinery of venom gland cells. In addition, a small number of transcripts were found to encode proteins that share sequence similarity with well-known venom constituents of social hymenopteran species, such as hyaluronidase-like proteins and an Allergen-5 protein. An overall number of 29 venom proteins could be identified through the combination of ESTs sequencing and proteomic analyses. The most highly redundant set of ESTs encoded a protein that shared sequence similarity with a venom protein of unknown function potentially specific of the Chelonus lineage. Venom components specific to C. inanitus included a C-type lectin domain containing protein, a chemosensory protein-like protein, a protein related to yellow-e3 and ten new proteins which shared no significant sequence similarity with known sequences. In addition, several venom proteins potentially able to interact with chitin were also identified including a chitinase, an imaginal disc growth factor-like protein and two putative mucin-like peritrophins. Conclusions The use of the combined approaches has allowed to discriminate between cellular and truly venom proteins. The venom of C. inanitus appears as a mixture of conserved venom components and of potentially lineage-specific proteins. These new molecular data enrich our knowledge on parasitoid venoms and more generally, might contribute to a better understanding of the evolution and functional diversity of venom proteins within Hymenoptera.

Early detection of subclinical organ dysfunction by microdialysis of the rectus abdominis muscle in a porcine model of critical intra-abdominal hypertension

The aim of this study was to evaluate microdialysis of the rectus abdominis muscle (RAM) for early detection of subclinical organ dysfunction in a porcine model of critical intra-abdominal hypertension (IAH). Microdialysis catheters for analyses of lactate, pyruvate, and glycerol levels were placed in cervical muscles (control), gastric and jejunal wall, liver, kidney, and RAM of 30 anesthetized mechanically ventilated pigs. Catheters for venous lactate and interleukin 6 samples were placed in the jugular, portal, and femoral vein. Intra-abdominal pressure (IAP) was increased to 20 mmHg (IAH20 group, n = 10) and 30 mmHg (IAH30, n = 10) for 6 h by controlled CO2 insufflation, whereas sham animals (n = 10) exhibited a physiological IAP. In contrast to 20 mmHg, an IAH of 30 mmHg induced pathophysiological alterations consistent with an abdominal compartment syndrome. Microdialysis showed significant increase in the lactate/pyruvate ratio in the RAM of the IAH20 group after 6 h. In the IAH30 group, the strongest increase in lactate/pyruvate ratio was detected in the RAM and less pronounced in the liver and gastric wall. Glycerol increased in the RAM only. After 6 h, there was a significant increase in venous interleukin 6 of the IAH30 group compared with baseline. Venous lactate was increased compared with baseline and shams in the femoral vein of the IAH30 group only. Intra-abdominal pressure-induced ischemic metabolic changes are detected more rapidly and pronounced by microdialysis of the RAM when compared with intra-abdominal organs. Thus, the RAM represents an important and easily accessible site for the early detection of subclinical organ dysfunction during critical IAH.