How the 'kitome' influences the characterization of bacterial communities in lepidopteran samples with low bacterial biomass.

AIMS: We aimed to elucidate whether the DNA extraction kit and bacteria therein affect the characterization of bacterial communities associated with butterfly samples harbouring different bacterial abundancies. METHODS AND RESULTS: We analysed bacteria associated with eggs of Pieris brassicae and with adults of this butterfly, which were either untreated or treated with antibiotics (ABs). Three DNA extraction kits were used. Regardless of the extraction kit used, PCR amplification of the bacterial 16S rRNA gene detected very low bacterial presence in eggs and AB-treated butterflies. In untreated butterflies, bacterial signal intensity varied according to the kit and primers used. Sequencing (MiSeq) of the bacterial communities in untreated and AB-treated butterflies revealed a low alpha diversity in untreated butterflies because of the dominance of few bacteria genera, which were detectable regardless of the kit. However, a significantly greater alpha diversity was found in AB-treated butterflies, evidencing a true bias of the results due to bacterial contaminants in the kit. CONCLUSIONS: The so-called 'kitome' can impact the profiling of Lepidoptera-associated bacteria in samples with low bacterial biomass. SIGNIFICANCE AND IMPACT OF THE STUDY: Our study highlights the necessity of method testing and analysis of negative controls when investigating Lepidoptera-associated bacterial communities.

Successful use of a percutaneous miniaturized extracorporeal life support system as a bridge and assistance to left ventricular assist device implantation in a patient with severe refractory cardiogenic shock.

We present a 51-year-old man with cardiogenic shock in whom a percutaneous extracorporeal life support system (ECLS) was inserted to restore cardiopulmonary stability. After successful stabilization, a left ventricular assist device was implanted, using the ECLS without switching to a conventional cardiopulmonary bypass system to reduce its side effects.

Out-of-hospital extracorporeal life support for cardiac arrest-A case report.

We report the use of out-of-hospital extracorporeal life support (ECLS) in cardiac arrest. We treated a 9-year-old girl with cardiac arrest after warm-water drowning with percutaneous venoarterial extracorporeal membrane oxygenation (ECMO) using a new portable Mini-ECMO system. A beating-heart circulation was reestablished on ECMO, but, unfortunately, our patient did not survive. This case shows that Mini-ECMO support can be used to restore an effective circulation and gas exchange in the out-of-hospital setting.

Percutaneous cannulation for extracorporeal life support.

BACKGROUND: Extracorporeal life-support systems are valuable tools to treat patients with acute cardiopulmonary failure in intensive care facilities, and are highly suitable for the interhospital transfer of critically ill patients to specialized centers. This article reviews the cannulation strategies and associated vascular complications in our institution. METHODS: Between January 2004 and December 2009, 464 extracorporeal life-support systems were implanted via percutaneous cannulation at our institution. The type and incidence of adverse events related to the percutaneous access to femoral, subclavian vessels and the jugular vein were retrospectively analyzed. The primary focus was on bleeding and limb ischemia. RESULTS: 464 patients (340 male, 124 female) with isolated pulmonary or combined cardiopulmonary failure were connected to extracorporeal gas exchange systems. Most patients (n = 196) were connected to a PECLA system; 158 patients to a veno-arterial ECMO. Use of a veno-venous ECMO system was necessary in 110 cases. Thirty-two patients (6.9 %) suffered bleeding complications after cannula insertion, predominantly after PECLA placement (3.9 %). After implantation, limb ischemia developed in 15 cases (3.2 %), mostly in the veno-arterial ECMO group (n = 13). Demographic data and cannula size show no significant difference between patient groups with and without ischemic complications ( P = 0.57). A prophylactic fasciotomy was performed in the 15 cases with limb ischemia. Survival was independent of ischemic (leg) complications. CONCLUSION: With proper vessel visualization, exposure and cannulation, and accurate cannula placement, optimal flows and minimal complication rates can be achieved, rendering percutaneous extracorporeal life support a safe procedure.

Extracorporeal life support - systems, indications, and limitations.

Extracorporeal life support is becoming increasingly popular with the miniaturization of the devices necessary for cardiac and pulmonary support. In the present report, the current systems, indications for placement and limitations are reviewed.

Innate immunity: eggs of Manduca sexta are able to respond to parasitism by Trichogramma evanescens.

So far, it was unknown whether immune responses of insect eggs are inducible or suppressed by parasitism. We investigated whether transcription of immune related genes in eggs of Manduca sexta changed in response to parasitism by Trichogramma evanescens. First, using DDRT-PCR, several cDNA elements known to represent immune related M. sexta genes inducible by bacterial challenge were isolated from eggs. In addition, two novel cDNAs were found: (a) immulectin-V (IML-V) suggested to be involved in recognition of foreign bodies, and (b) a new like-moricin protein with possible antimicrobial effects (L-Mor). Quantitative real time RT-PCR analyses revealed enhanced transcription in parasitized eggs compared to unparasitized ones for IML-V, prophenoloxidase (ProPO), prophenoloxidase activating protease I (PAP I), and proparalytic peptide (ProPP). No significant differences between parasitized and unparasitized eggs were detected for sequences encoding the antimicrobial peptides L-Mor, leureptin Leu, and attacin II Att II. Transcript levels of other antibacterial peptides were suppressed after parasitization for 3d (cecropin 6, Cec 6) and 2d (gloverin, Glov). While nearly 100% of the Manduca eggs contained Trichogramma specimens 1d after exposure to parasitoids, only 64% of the host eggs harbored parasitoid larvae 4d after parasitization. Our data demonstrate that the immune system of Manduca eggs shows differentiated responses to parasitization and suggest that insect eggs can defend against parasitization.

A class of compact dwarf galaxies from disruptive processes in galaxy clusters.

Dwarf galaxies have attracted increased attention in recent years, because of their susceptibility to galaxy transformation processes within rich galaxy clusters. Direct evidence for these processes, however, has been difficult to obtain, with a small number of diffuse light trails and intra-cluster stars being the only signs of galaxy disruption. Furthermore, our current knowledge of dwarf galaxy populations may be very incomplete, because traditional galaxy surveys are insensitive to extremely diffuse or compact galaxies. Aware of these concerns, we recently undertook an all-object survey of the Fornax galaxy cluster. This revealed a new population of compact members, overlooked in previous conventional surveys. Here we demonstrate that these 'ultra-compact' dwarf galaxies are structurally and dynamically distinct from both globular star clusters and known types of dwarf galaxy, and thus represent a new class of dwarf galaxy. Our data are consistent with the interpretation that these are the remnant nuclei of disrupted dwarf galaxies, making them an easily observed tracer of galaxy disruption.

Rapamycin reduces neointima formation during vascular injury.

BACKGROUND: Proliferation and migration of vascular smooth muscle cells (SMCs) mark the key processes in the development of bypass graft disease and during neointima formation in restenosis after angioplasty. Growth factors are potent SMC mitogens as they are involved in SMC proliferation and in extracellular matrix (ECM) synthesis. Based on these premises, we examined the effect of the proliferation inhibitor rapamycin in human SMC culture and in a rabbit vascular injury model. MATERIALS AND METHODS: Injection of rapamycin or its vehicle was performed with an infusion-balloon catheter directly into the vessel wall during vascular injury. The intima/media ratio was determined histologically whereas the protein expression was analysed using the powerful two-dimensional gel electrophoresis (2D page) technique. Inhibition of proliferation after rapamycin application was estimated in a human SMC culture for time and dose dependent effects. RESULTS: Rapamycin treatment resulted in a significant reduction of intima media ratio compared to vehicle treated animals after three weeks (0.65 +/- 0.1 vs. 1.2 +/- 0.2 intima-media-ratio, p < 0.05). 2D electrophoresis analysis proved increased ECM synthesis following angioplasty (i.e., lamin, vimentin) in vehicle treated animals. Local rapamycin administration resulted in profound reduction of ECM synthesis after vascular injury. In in-vitro experiments exposure of cultured human SMCs to rapamycin resulted in a significant and dose-dependent (1 nm-100 nm) reduction of human smooth muscle cell proliferation measured by cell counting. CONCLUSION: These above mentioned results suggest that protein synthesis in addition to reduction of cellular proliferation plays an important role following vascular injury, since application of rapamycin resulted in the reduction of SMC proliferation and ECM-synthesis.

A review of 507 off-pump coronary bypass patients: a single center experience.

This retrospective study evaluates perioperative results of 'Off-pump' coronary artery bypass surgery (OPCAB) experience in a single center. Five hundred and seven patients were operated (median sternotomy) from 1998 to 2002 using OPCAB. Patient data were registered and risk prediction calculated using the full logistic version of EuroSCORE. Overall, 1091 distal anastomoses were performed and only five (1%) patients required conversion to cardiopulmonary bypass. The predicted mortality was 3.8+/-4.5%, the observed mortality was 2.37%. OPCAB technique has become a standard approach in our department with low mortality and morbidity rates even in treatment of multivessel disease or high-risk patients.

Transfection of E-cadherin cDNA in human lung tumor cells reduces invasive potential of tumors.

Lung cancer is a major health-care problem in industrialized countries. With reference to its therapeutic consequences and major histological variations, it is divided into two subgroups - SCLC (small-cell lung cancer) and NSCLC (non-small-cell lung cancer). As an important factor of cell-cell and cell-substratum interaction, cell adhesion molecules (CAMs) seem to play a key role in tumor-cell migration and invasion that lead to metastases. We investigated human lung tumor cell lines established from histologically documented neoplastic lesions taken in our operating theater. Immunohistological screening showed differences in E-cadherin expression with no clear predominance of SCLC or NSCLC cell lines. Using an invasion model with Matrigel Matrix and a migration assay, we could demonstrate a more aggressive behavior pattern in E-cadherin-negative cell lines. We transfected E-cadherin cDNA into a formerly negative cell line showing strong invasive behavior in the initial tests in order to investigate the role of E-cadherin in this process. In this study, we examined E-cadherin cDNA transfection in human bronchial carcinoma cells. At present, transfection is stable with a follow-up time of one year. We could demonstrate that cell lines were remarkably less invasive after transfection of E-cadherin in the invasion model with Matrigel Matrix. These results indicate that the E-cadherin CAM plays an important role in lung tumor invasion and metastasis. Further studies are in progress to confirm these findings and to describe a possible role of this CAM in tumor therapy.

Bypass graft disease: analysis of proliferative activity in human aorto-coronary bypass grafts.

BACKGROUND: Aortocoronary bypass graft disease with its increasing clinical signification represents an unsolved problem in cardiological and heart surgery practice. Late occlusion of autologous saphenous vein grafts occurs against a background of medial and neointimal thickening due to migration and proliferation of smooth muscle cells and the later appearance of atherosclerotic plaques. To clarify the role of cellular proliferation in humans we characterized the cellular composition and proliferative index in 30 stenotic saphenous vein grafts. METHODS: 30 stenotic vein grafts and 25 control veins were explantated during redo heart surgery procedures. Time between initial surgical intervention and explantation was 3-168 month (mean 94,8 month). The total area and cell count of the neointima, media and adventitia was calculated computer assisted. Actively proliferating cells were identified using antibody to Ki-67 and by double-lable immuncytochemistry with alpha SMC actin, CD 31 (endothelial cells), CD 68 lpar;makrophages) and CD 45 (T-lymphocytes). RESULTS: Active proliferation was detected in different cell typs with a mean proliferation index of 0.15%,0.18% and 0.086% for the neointima, media and adventita. Only 9% of the proliferating cells in the neointima were SMC (not identified cells 40%); corresponding 14% SMC (not identified cells 33%) were detected in the media. Endothelial cells were the predominante proliferating cell type in all sites of the vessel wall. CONCLUSION: 1. Proliferation occured at low level. While proliferation may play an important role in early lesions our data imply low proliferation activity in advanced graft lesions. Other mechanism like production and deposition of extracellular matrix (ECM) in the neointima are responsible for the lumen reduction of bypass grafts. 2. The high portion of unidentified cells may represent SMC or other cell types at different stages of differentiation; this requires further investigation. 3. The identification of proliferating macrophages and T-lymphocytes implicate an inflammatory component in the development of human bypass graft lesions.

[Treatment of supravalvular pulmonary stenosis after arterial switch operations (ASO)]. / Die Behandlung supravalvulärer Pulmonalstenosen nach Arteriellen Switch-Operationen (ASO).

The aim of this study was to evaluate the incidence of postoperative pulmonary supravalvular stenosis in patients with d-TGA and to assess the rate of success or failure of balloon angioplasty. Out of 70 patients with d-TGA 67 patients underwent successful arterial switch operation. Twelve children developed severe supravalvular pulmonary stenosis with a peak gradient above 50 mmHg (range: 50-120 mmHg). In these patients 19 high pressure dilatations were performed up to a diameter of 130% of the native valve dimension. The mean age at angioplasty was 17 months (range: 3-36 months). Successful intervention was defined as a > 50% decrease of predilatation peak pressure gradient or right ventricular pressure < 50 mmHg. Dilatations were performed without complications. Complete resolution was primarily achieved in 1 patient. In 7 patients the pressure gradients could be reduced to 10-45 mmHg (mean: 25 mmHg). In another two patients a palliative stent-implantation into the pulmonary trunk was necessary to reduce the pressure gradient. Because unsuccessful intervention, two patients needed subsequent operation. During follow-up of 6-9 months after intervention severe restenosis occurred in 3 patients (2 after stent-implantation; 1 after re-re-dilatation) who then also needed operation. Balloon dilatation should be the first treatment in patients with pulmonary stenosis after ASO in TGA owing to the low complication rate and the potential benefit of this procedure. Recurrent and combined stenoses with narrow pulmonary valve annulus should be treated surgically.

Analysis of volatiles induced by oviposition of elm leaf beetle Xanthogaleruca luteola on Ulmus minor.

Egg deposition of the elm leaf beetle Xanthogaleruca luteola causes the emission of volatiles from its food plant, Ulmus minor. These volatiles are exploited by the egg parasitoid, Oomyzus gallerucae, to locate its host. In contrast to other tritrophic systems, the release of volatiles is not induced by feeding but by egg deposition. Previous investigations showed that the release is systemic and can be triggered by jasmonic acid. Comparison of headspace analysis revealed similarities in the blend of volatiles emitted following egg deposition and feeding. The mixture consists of more than 40 compounds; most of the substances are terpenoids. Leaves next to those carrying eggs emit fewer compounds. When treated with jasmonic acid, leaves emit a blend that consists almost exclusively of terpenoids. Dichloromethane extracts of leaves treated with jasmonic acid were also investigated. After separation of extracts of jasmonate induced elm leaves on silica, we obtained a fraction of terpenoid hydrocarbons that was attractive to the parasitoids. This indicates that jasmonic acid stimulates the production of terpenoid hydrocarbons that convey information of egg deposition to the parasitoid.

Host finding and oviposition behavior in a chrysomelid specialist--the importance of host plant surface waxes.

Although several studies have elucidated the role of plant epicuticular waxes in host recognition and oviposition by herbivorous insects, there is little known about this subject in Chrysomelidae. In the present study, chemical cues for host recognition behavior and oviposition by the monophagous chrysomelid species Cassida stigmatica were investigated with special regard to surface waxes of the host plant. Tanacetum vulgare (Asteraceae). After hibernation in the soil, adults of this species must climb the plant's petioles, which emerge from the ground in spring. The response of adult C. stigmatica to contact and volatile cues from petioles and leaves of T. vulgare was investigated in a "stem arena," in which differently treated petioles and petiole dummies were offered. Volatile and contact cues of T. vulgare petioles served as well for host recognition. The contact cues were isolated from the petioles and leaves by hexane extraction and by cellulose acetate treatment, which removed cuticular waxes. The attractive volatiles were not extracted sufficiently by hexane. To examine the role of cuticular waxes of the host leaf surface in oviposition, female C. stigmatica were offered intact leaflets and leaflets from which cuticular waxes had been stripped by cellulose acetate treatment. Females did not discriminate between intact and stripped leaflets when only the upper leaf surfaces were offered. However. when the lower leaf surfaces that are generally used as oviposition sites were offered, C. stigmatica preferred to lay eggs on intact leaflets. We conclude that waxes of the lower leaf surface contain crucial information for oviposition in C. stigmatica.

Aortic valve preservation in acute type A dissection: mid-term results.

BACKGROUND AND AIM OF THE STUDY: The study aim was to evaluate the long-term effectiveness of a strategy for managing the aortic valve, aortic root and ascending aorta according to the pathology in acute aortic type A dissection. Results after surgery for acute type A dissection with preservation of the aortic valve were reviewed. METHODS: The patient group included 57 hospital survivors operated on according to a surgical strategy of aortic valve resuspension and supracoronary ascending aortic graft implantation. Reinforcement of the aortic stumps with gelatin-resorcinol-formaldehyde glue was performed in all patients. Aortic valve function in all survivors was investigated by echocardiographic follow up at 30 days, 6 and 12 months after surgery, and yearly thereafter. RESULTS: During the follow up period, nine patients (16%) died without reoperation. Actuarial probability of freedom from reoperation for aortic valve failure in the complete series was estimated as 100% after both 30 days and 12 months. Postoperatively, one patient underwent reoperation 14 months for aortic regurgitation, and three patients for aortic regurgitation with sinus of Valsalva dilatation between 48 and 88 months. The hospital mortality rate at reoperation was 50% (n = 2). CONCLUSION: Valve-sparing surgery is possible and can be recommended for the majority of patients with acute type A aortic dissection.

Expression of the proto-oncogene c-myc in human stenotic aortocoronary bypass grafts.

Proliferation and differentiation of vascular smooth muscle cells (VSMC) are central events in vascular pathobiology and play a major role in the development of stenotic and restenotic lesions. The proto-oncogene c-myc and other early cell cycle-regulating genes have been implicated in the induction of cell proliferation and differentiation under diverse pathophysiological conditions. In the present study we analyzed c-myc mRNA expression by indirect nonradioactive in situ hybridization technique (NISH) in human stenotic venous bypass grafts (n = 32) retrieved during re-do operations of coronary artery disease and compared the results with 28 native veins (vena saphena magna) from the same patients. Stenotic bypass grafts showed enhanced c-myc expression located predominantly in VSMC in the media and neointima (severity score: ++-+++, 32/32 stenotic veins). In native veins we observed only low levels of c-myc mRNA (severity score: +, 28/28 native veins), all signals were restricted to endothelial cells of either the innermost intimal layer or of the vasa vasorum. Our in situ hybridization studies demonstrate enhanced mRNA expression of the proto-oncogene c-myc in stenotic venous bypass grafts. These results suggest that--in analogy to other pathophysiological conditions--c-myc exerts essential regulatory functions in cellular events operative during the initiation and progression of venous bypass graft disease.

The effect of a green leaf volatile on host plant finding by larvae of a herbivorous insect.

The role of a general green leaf volatile (glv) in host finding by larvae of the oligophagous chrysomelid Cassida denticollis was investigated using a new bioassay which takes into account the need for neonate larvae of this species to climb fresh host plants from the ground. A "stem arena" was designed in which plant stems of the host, tansy (Tanacetum vulgare), and stem dummies (tooth picks), both wrapped in perforated filter paper, were offered to neonate larvae. The wrapping allowed olfactory responses to be tested by preventing access to contact stimuli of stems and dummies. Larvae significantly preferred to climb the wrapped tansy stems over dummies after a period of 15 min. The test glv, (Z)-3-hexen-1-ol, was not attractive when applied to dummies. However, when the glv was applied to the bottom of the arena, the ability of larvae to discriminate between host stems and untreated dummies was significantly enhanced. More larvae climbed wrapped host stems than dummies even within 5 min. While numerous other herbivorous insects are known to be directly attracted by glv, this study shows that a singly offered glv on its own is unattractive to an herbivore but enhances the herbivore's ability to differentiate between host and nonhost plants.

Cardiac pacing in premature infants and neonates: steroid eluting leads and automatic output adaptation.

BACKGROUND: Appropriate generator and lead selection as well as techniques of implantation are most important aspects of cardiac pacing in the extremely young patient. Here we report the clinical results using a new technique with automatic output adaptation based on evoked response in combination with steroid-eluting epicardial leads in small children. METHODS: One neonate and 2 premature infants underwent permanent pacemaker implantation because of congenital high-degree atrioventricular block or postoperative complete heart block, respectively. Steroid-eluting epicardial leads and a multiprogrammable pacemaker with automatic output adaptation were used. RESULTS: Intermuscular abdominal generator placement and epicardial suture-fixation of the bipolar lead through a subcostal approach was without complications. Serial follow-up examinations revealed safe and consistent pacemaker function up to 12 months after operation. CONCLUSIONS: The technique represents an excellent alternative for permanent cardiac pacing in extremely small patients. We believe that it provides an increase in functional lifetime of the devices and delays the need for battery replacement with its associated complications in this young patient population.