Propofol sedation in Drug Induced Sedation Endoscopy without an anaesthesiologist â€" a study of safety and feasibility.

BACKGROUND: Propofol sedation in Drug Induced Sedation Endoscopy (DISE) of the upper airway of patients with obstructive sleep apnea (OSA) without the presence of anesthesiologist has not been done before. Propofol sedation is normally administered by an anesthesiologist. This is the first study of this new method. METHODOLOGY: Based on the positive experience with Nurse-administered Propofol Sedation (NAPS) for endoscopic procedures in the departments of gastroenterology we wanted to test the set-up as method of propofol sedation for DISE procedures in our Otorhinolaryngology (ORL) department. The ORL specialists and staff nurses that carry out DISE procedures all underwent a formalized education in Nurse-administered Propofol Sedation before the study. We included 200 patients with severe snoring and / or obstructive sleep apnea. They were referred for DISE examination prior to possible targeted surgery based on the findings. RESULTS: In our study the aforementioned ORL team successfully cared out propofol sedation without the presence of an anesthesiologist. All examinations were carried out according to plan. There were no adverse events during the procedures or in the following observational period. CONCLUSIONS: The NAPS method of sedation for DISE seems safe and feasible when performed by trained staff in a hospital setting.

Human babesiosis in Europe: what clinicians need to know.

Although best known as an animal disease, human babesiosis is attracting increasing attention as a worldwide emerging zoonosis. Humans are commonly infected by the bite of ixodid ticks. Rare ways of transmission are transplacental, perinatal and transfusion-associated. Infection of the human host can cause a very severe host-mediated pathology including fever, and hemolysis leading to anemia, hyperbilirubinuria, hemoglobinuria and possible organ failure. In recent years, apparently owing to increased medical awareness and better diagnostic methods, the number of reported cases in humans is rising steadily worldwide. Hitherto unknown zoonotic Babesia spp. are now being reported from geographic areas where babesiosis was not previously known to occur and the growing numbers of travelers and immunocompromised individuals suggest that the frequency of cases in Europe will also continue to rise. Our review is intended to provide clinicians with practical information on the clinical management of this rare, but potentially life-threatening zoonotic disease. It covers epidemiology, phylogeny, diagnostics and treatment of human babesiosis and the potential risk of transfusion-transmitted disease with a special focus on the European situation.

The hormone melatonin stimulates renoprotective effects of "early outgrowth" endothelial progenitor cells in acute ischemic kidney injury.

Endothelial progenitor cells (EPCs) protect the kidney from acute ischemic injury. The aim of this study was to analyze whether pretreatment of murine "early outgrowth" EPCs (eEPCs) with the hormone melatonin increases the cells' renoprotective effects in the setting of murine acute ischemic renal failure. Male (8-12 wk old) C57Bl/6N mice were subjected to unilateral ischemia-reperfusion injury postuninephrectomy (40 min). Postischemic animals were injected with either 0.5×10(6) untreated syngeneic murine eEPCs or with cells, pretreated with melatonin for 1 h. Injections were performed shortly after reperfusion of the kidney. While animals injected with untreated cells developed acute renal failure, eEPC pretreatment with melatonin dramatically improved renoprotective actions of the cells. These effects were completely reversed after cell pretreatment with melatonin and the MT-1/-2 antagonist luzindole. In vitro analysis revealed that melatonin reduced the amount of tumor growth factor-ß-induced eEPC apoptosis/necrosis. Secretion of vascular endothelial growth factor by the cells was markedly stimulated by the hormone. In addition, migratory activity of eEPCs was enhanced by melatonin and supernatant from melatonin-treated eEPCs stimulated migration of cultured mature endothelial cells. In summary, melatonin was identified as a new agonist of eEPCs in acute ischemic kidney injury.

Established and emerging pathogens in Ixodes ricinus ticks collected from birds on a conservation island in the Baltic Sea.

Tick-borne pathogens such as Lyme borreliosis spirochaetes, Anaplasma phagocytophilum, Rickettsia spp. and Babesia spp. cause a great variety of diseases in animals and humans. Although their importance with respect to emerging human diseases is increasing, many issues about their ecology are still unclear. In spring 2007, 191 Ixodes ricinus (Acari: Ixodidae) ticks were collected from 99 birds of 11 species on a bird conservation island in the Baltic Sea in order to test them for Borrelia spp., A. phagocytophilum, Rickettsia spp. and Babesia spp. infections. Identification of the pathogens was performed by polymerase chain reaction (PCR), restriction fragment length polymorphism and sequence analysis. The majority of birds with ticks testing positive were European robins and thrushes. Borrelia DNA was detected in 14.1%, A. phagocytophilum in 2.6%, rickettsiae in 7.3% and Babesia spp. in 4.7% of the ticks. Co-infections with different pathogens occurred in six ticks (3.1%). The fact that 11 ticks (five larvae, six nymphs) were infected with Borrelia afzelii suggests that birds may, contrary to current opinion, serve as reservoir hosts for this species. Among rickettsial infections, we identified Rickettsia monacensis and Rickettsia helvetica. As we detected five Rickettsia spp. positive larvae and two birds carried more than one infected tick, transmission of those pathogens from birds to ticks appears possible. Further characterization of Babesia infections revealed Babesia divergens and Babesia microti. The occurrence of Babesia spp. in a total of five larvae suggests that birds may be able to infect ticks, at least with Ba. microti, a species considered not to be transmitted transovarially in ticks.

Growth in microculture of single tobacco cells infected with tobacco mosaic virus.

Single tobacco callus cells with or without tobacco mosaic virus inclusion bodies from systemically infected Nicotiana tabacum plants were grown in microcultures. The culture medium consisted of mineral salts and sucrose; it also contained coconut milk. Out of 100 inclusion-bearing cells and 150 inclusion-free cells, 10 and 70 cultured cells divided; eventually 5 and 65 cells, respectively, formed single cell clones. The 5 clones derived from inclusion-bearing cells, and all but 3 of 40 clones from inclusion-free cells, showed virus inclusions in somne cells. The virus could not be detected in three inclusion-free clones by local lesion assay. The results suggest single-cell culture methods for differentiating virus-free plants from cells of pathogen-infected plants.

Babesiosis: recent insights into an ancient disease.

Ever since the discovery of parasitic inclusions in erythrocytes of cattle in Romania by Victor Babes at the end of the 19th century, newly recognised babesial pathogens continue to emerge around the world and the substantial public health impact of babesiosis on livestock and man is ongoing. Babesia are transmitted by ixodid ticks and infection of the host causes a host-mediated pathology and erythrocyte lysis, resulting in anemia, hyperbilirubinuria, hemoglobinuria, and possibly organ failure. Recently obtained molecular data, particularly for the 18S rRNA gene, has contributed significantly to a better understanding of the sometimes puzzling phylogenetic situation of the genus Babesia and new information has been added to help determine the taxonomic position of many species. Moreover, it seems that owing to higher medical awareness the number of reported cases in humans is rising steadily. Hitherto unknown zoonotic babesias are now being reported from geographical areas where babesiosis was not known to occur and the growing numbers of immunocompromised individuals suggest that the frequency of cases will continue to rise. This review covers recent insights into human babesiosis with regard to phylogeny, diagnostics and treatment in order to provide new information on well known as well as recently discovered parasites with zoonotic potential.

Nitric oxide reduces organ injury and enhances regeneration of reduced-size livers by increasing hepatic arterial flow.

BACKGROUND: Reduced-size livers suffer from portal hyperperfusion, diminished arterial blood flow and the risk of postoperative liver injury. The aim of this experimental study was to unravel the role of nitric oxide in this setting. METHODS: Rats underwent 85 per cent partial hepatectomy and either substitution of nitric oxide with molsidomine or inhibition of nitric oxide synthase (NOS) with N(G)-nitro-L-arginine methyl ester. Untreated hepatectomized animals served as controls and unresected animals as the sham group. RESULTS: Ultrasonic flowmetry following partial hepatectomy revealed a marked increase in portal venous inflow with a concomitant decrease in hepatic arterial inflow. Nitric oxide substitution counteracted the decline in hepatic arterial inflow and caused a significantly greater increase in cell proliferation after partial hepatectomy compared with control or NOS-inhibited animals. Hepatectomized animals further profited from nitric oxide substitution, as indicated by reduced aminotransferase release and improved liver function. CONCLUSION: Nitric oxide improves the postoperative course of rats with reduced-size livers by modulating hepatic macrohaemodynamics and mediating regeneration and cytoprotection, but not by reducing hepatic hyperperfusion and the accompanying sinusoidal shear stress.

Optical immunosensor for fast and sensitive detection of DDT and related compounds in river water samples.

A surface plasmon resonance (SPR) based immunosensor has been developed for the monitoring of environmentally persistent pollutants like DDT, its metabolites and analogues in real water samples. A reusable immunosurface is provided via the covalent attachment of the analyte derivative to a self-assembled alkanethiol monolayer formed onto the SPR gold-thin layer. The regeneration of the sensor surface allowed the performance of 270 assay cycles within an analysis time of 20 min for each assay cycle. Immunoassays based on a binding inhibition format were performed by using two monoclonal antibodies (MAbs) with different selectivity. Low limits of detection (LODs), in the sub-nanogram per litre range, were attained for DDT-selective (15 ng L-1) and DDT group-selective immunoassays (31 ng L-1). Both assays were carried out in spiked river water samples without significant effect of the matrix. SPR measurements were validated using gas-chromatography-mass spectrometry. The comparison between methods was in good agreement showing an excellent correlation coefficient (r2=0.995). The SPR analysis of DDT proved to be three times more sensitive than colorimetric ELISAs without the need of labelling and a much lower time of response. Our SPR biosensor portable platform (beta-SPR) is already commercialised by the company SENSIA, S.L. (Spain).

A novel transcriptome subtraction method for the detection of differentially expressed genes in highly complex eukaryotes.

We have designed a novel transcriptome subtraction method for the genome-scale analysis of differential gene expression in highly complex eukaryotes, in which suppression subtractive hybridization (SSH) is performed first to enrich the target and, after exchange of adapters, negative subtraction chain (NSC) is then used to eliminate the remaining background. NSC evolved from differential subtraction chain (DSC). We designed novel adapters which make the subtraction system more robust. SSH and NSC were then combined to successfully detect differentially expressed genes in Solanum. The combined technique improves qualitatively upon SSH, the only commercially available transcriptome subtraction system, by detecting target genes in the middle abundance class, to which most differentially expressed genes in highly complex eukaryotes are expected to belong. The main advantage of the combined technique with SSH/NSC is its ability to isolate differentially expressed genes quickly and cost-efficiently from non-standard models, for those microarrays are unavailable.

Determination of carbaryl in natural water samples by a surface plasmon resonance flow-through immunosensor.

The analysis of carbaryl in natural water samples was accomplished using a portable immunosensor based on surface plasmon resonance (SPR) technology. The assay was based on a binding inhibition immunoassay format with the analyte derivative covalently immobilized on the sensor surface. An alkanethiol self-assembled monolayer (SAM) was formed onto the gold-coated sensor surface to allow the reusability of the same sensing surface during 220 regeneration cycles. Reproducibility was evaluated by performing three independent assays in triplicate on 3 different days. The batch-assay variability was also calculated using three different gold-coated sensor surfaces. The intra- and inter-day relative standard deviation were 8.6 and 15.3%, respectively, whilst a variation of 7.4% in assay sensitivity was obtained by employing different sensor chips. The lowest detection limit, calculated as the concentration providing a 10% decrease of the blank signal, was of 1.38 microg L(-1). Matrix effects were also evaluated in different water types, showing I50 values (carbaryl concentrations that produced a 50% decrease of the blank signal) within the range of carbaryl standard curves in distilled water (2.78-3.55 microg L(-1)). The carbaryl immunoassay performance was validated with respect to conventional high-performance liquid chromatography-mass spectrometry (HPLC-MS). The correlation between methods was in good agreement (r2 = 0.998, 0.999 and 0.999) for the three types of natural water samples tested. A complete assay cycle, including regeneration, is accomplished in 20 min. All measurements were carried out with the SPR sensor system (beta-SPR) commercialised by the company SENSIA, SL (Spain). The small size and low-time of response of the beta-SPR platform would allow its utilization in real contaminated locations.

Public knowledge and attitudes on organ donation do not differ in Germany and Spain.

BACKGROUND: The background of this study was the inadequate supply of donor organs in Germany. In Spain, by contrast, a strong increase of organ donors over the past years has created a satisfactory supply situation. Because both countries have similar legal situations, the causes for the drastic differences in organ donation rates remain unclear. The main issue of our study was to investigate the intellectual attitudes toward various aspects of postmortem donations in the populations of both countries as a causative factor for the observed differences. METHODS: We studied 726 persons by questionnaire. Probands, matched for age and gender, were recruited among medical students, in a public library and in a general medical practice in both Germany and Spain. RESULTS: We found no differences in the attitudes toward postmortem organ donation between the two countries. Differences among the social groups within the countries were apparent in the expected direction. CONCLUSION: A higher level of knowledge or a difference in attitudes toward organ donation is probably not the reason for the higher donation rate in Spain. The cause appears to be rather at the organizational level.

Face and emotion expression processing and the serotonin transporter polymorphism 5-HTTLPR/rs22531.

Face cognition, including face identity and facial expression processing, is a crucial component of socio-emotional abilities, characterizing humans as highest developed social beings. However, for these trait domains molecular genetic studies investigating gene-behavior associations based on well-founded phenotype definitions are still rare. We examined the relationship between 5-HTTLPR/rs25531 polymorphisms - related to serotonin-reuptake - and the ability to perceive and recognize faces and emotional expressions in human faces. For this aim we conducted structural equation modeling on data from 230 young adults, obtained by using a comprehensive, multivariate task battery with maximal effort tasks. By additionally modeling fluid intelligence and immediate and delayed memory factors, we aimed to address the discriminant relationships of the 5-HTTLPR/rs25531 polymorphisms with socio-emotional abilities. We found a robust association between the 5-HTTLPR/rs25531 polymorphism and facial emotion perception. Carriers of two long (L) alleles outperformed carriers of one or two S alleles. Weaker associations were present for face identity perception and memory for emotional facial expressions. There was no association between the 5-HTTLPR/rs25531 polymorphism and non-social abilities, demonstrating discriminant validity of the relationships. We discuss the implications and possible neural mechanisms underlying these novel findings.

A method for isolation of nuclei containing undegraded RNA from RNAase-rich plasmodia of Physarum polycephalum.

(1) In order to protect the nuclear RNA of Physarum polycephalum plasmodia during cell homogenisation and purification of the nuclei, the following conditions were used: low temperature (-11 degrees C), high pH (8.1-8.9), formaldehyde (2.8%) and spermine (2.3 mM). (2) The efficiency of these RNAase-inhibiting and inactivating conditions is demonstrated by the high molecular weight of the processing products of transcripts from ribosomal genes (11.9, 9.5 and 5.0 kilobases), which were recovered from the isolated nuclei and visualised on agarose gels. (3) Hybridisation experiments with a DNA probe from an actin gene on size-fractionated nuclear RNA (Northern blots) indicate that the transcripts from actin genes are rapidly spliced in P. polycephalum. (4) The nuclear polyadenylated RNA has an average size of about 2.2 kb, which is not significantly larger than the average length of mRNA.

A highly conserved repetitive sequence from Physarum polycephalum contains nucleotide arrangements similar to replicator sequences.

An interspersed repetitive sequence from Physarum polycephalum has been cloned and analysed. The 394 bp sequence is highly conserved and contains several homopolymeric (dA)-(dT) tracts capable of forming bent DNA structures and a 10/11 match to the yeast-ARS-consensus sequence. The repetition frequency of the described sequence is about 3000 to 7000, a number that would fit with the distribution of replicator segments in Physarum.

Levels of RNA polymerases during the mitotic cycle of Physarum polycephalum.

Two RNA polymerase activities were quantitatively solubilized in plasmodial homogenates from Physarum polycephalum by sonication at 0.5 M ammonium chloride concentration. The proportions of RNA polymerases A and B were determined by four different methods. Equal activity levels of both enzyme A and enzyme B were detected throughout the synchronous mitotic cycle of Physarum.

Decreased expression of ventricular angiotensin receptor type 1 mRNA after human heart transplantation.

Myocardial angiotensin receptors of type 1 (AT1) are downregulated at the protein and mRNA levels in human heart failure. No data are available for the transplanted human heart, which frequently exhibits functional alterations. The aim of the present study was the quantitation of ventricular AT1 mRNA content in endomyocardial biopsies from patients after heart transplantation. For the determination of AT1 mRNA we used a novel quantitative reverse transcription polymerase chain reaction with low variance (6%) based on an internal AT1 cRNA standard, liquid-phase hybridization of polymerase chain reaction products in microtiter plates, and quantitation by enzyme-linked immunosorbent assay. Right ventricular biopsies from 16 patients after heart transplantation (left ventricular ejection fraction 67 +/- 7%) were compared with 12 patients with normal cardiac function (left ventricular ejection fraction 62 +/- 5%). A 46% lower AT1 mRNA content was found in biopsies from the 16 patients after heart transplantation than in the 12 controls (heart transplantation, 200 +/- 25 AT1 mRNA copies/ng RNA; controls, 368 +/- 50; P < 0.01). When AT1 mRNA content was related to the stably expressed GAPDH mRNA, a 49% decrease was detected (AT1/GAPDH: patients, 2.4 +/- 0.25; controls, 4.7 +/- 0.6; P < 0.006). No association between the extent of AT1 downregulation and clinical or hemodynamic variables was detected. In the human heart ventricular AT1 is downregulated after orthotopic heart transplantation. The decrease in AT1 mRNA is not associated with altered systolic function. This may partially reflect a loss of autonomic nerves and thus altered nervous control of the heart.

Reduced atrial angiotensin receptor type 1 mRNA content in end-stage human heart failure: assessment by a novel quantitative PCR-ELISA technique.

The number of atrial angiotensin II binding sites is reduced in end-stage human heart failure. The goals of our study were the development of a quantitative polymerase chain reaction for angiotensin II receptor type 1 mRNA to determine the angiotensin receptor type 1 (AT1) mRNA content in the atria of patients with end-stage heart failure. We established a quantitative PCR based on coamplification of AT1 wild-type and an internal standard in the same PCR, followed by liquid-phase hybridization of PCR products in microtiter plates and quantitation by ELISA. Glyceraldehyde phosphate dehydrogenase mRNA in the same samples was used to relate the AT1 mRNA content to a stably expressed reference gene. Atrial samples from 11 patients with end-stage heart failure obtained at cardiac transplantation were compared with atrial samples from 11 patients with normal cardiac function undergoing routine cardiac surgery. A PCR/ELISA system with a variance of about 6% after reverse transcription and a linear measuring range was established. In the samples from 11 patients with end-stage heart failure a 58% decrease in AT1 mRNA content was found in comparison with 11 controls (heart failure: 185,680 +/- 196,912 AT1 mRNA copies/microgram RNA, controls: 440,555 +/- 268,456, P < 0.02). When AT1 mRNA content was related to glyceraldehyde phosphate dehydrogenase mRNA, a 65% decrease was detected (AT1/glyceraldehyde phosphate dehydrogenase: heart failure: 4.84 +/- 5.18; controls: 13.74 +/- 7.77; P < 0.005). Standardization of PCR resulting in a low coefficient of variance, high reproducibility, and large sample capacity is possible using optimal internal standardization and the liquid-phase hybridization/ELISA system for detection. The optimized PCR procedure indicated downregulation of atrial AT1 in end-stage human heart failure, suggesting a reduced capacity of the atria to respond to angiotensin II stimulation in end-stage heart failure.

Angiotensin receptor type 1 mRNA in human right ventricular endomyocardial biopsies: downregulation in heart failure.

OBJECTIVE: Atrial angiotensin II receptors type 1 (AT1) are downregulated in end-stage human heart failure at mRNA and protein level. The present study investigated whether AT1 ventricular mRNA content was reduced in myocardial biopsies from heart failure patients. METHODS: AT1 mRNA was quantitated in right ventricular endomyocardial biopsies from 16 patients with decreased left ventricular function (LVEF 36 +/- 3%) due to dilated cardiomyopathy (DCM) and in biopsies from 12 patients with suspected myocardial disease but normal cardiac function (LVEF 62 +/- 2%). Two biopsies per patient were pooled, RNA was extracted and reverse-transcribed after addition of an AT1 cRNA standard. AT1 standard and wild-type RNA were amplified with the same primers in the same PCR tube. The PCR products were hybridized to a microtiter plate and detected and quantitated by an ELISA system. Glyceraldehyde phosphate dehydrogenase (GAPDH) mRNA was determined in the same samples as AT1 mRNA. RESULTS: In the biopsies from 16 patients with heart failure, a 68% decrease in AT1 mRNA content was found in comparison with 12 controls (heart failure 94 +/- 15 AT1 mRNA copies/ng RNA; controls 297 +/- 45; P < 0.001). Relating AT1 mRNA content to GAPDH mRNA confirmed the specific decrease in AT1 mRNA (AT1/GAPDH: heart failure 1.3 +/- 0.15; controls 3.4 +/- 0.5; p < 0.002). The best correlation between AT1 mRNA content and clinical parameters was found for right ventricular ejection fraction (r = 0.59, P < 0.01). CONCLUSIONS: The quantitative RT-PCR procedure indicated a loss of ventricular AT1 mRNA in human heart failure which corresponds to the loss of AT1 protein described previously. It may underlie the decrease in AT1 protein expression in human heart failure.

15-Deoxyspergualin for induction of graft nonreactivity after cardiac and renal allotransplantation in primates.

In order to assess the immunosuppressive potentials of 15-deoxyspergualin (15-DS) in a preclinical experiment, heterotopic cardiac (n = 27, group I) and classic renal (n = 25, group II) allotransplantations were performed in Chacma baboons. The following immunosuppressive regimens were applied: Groups IB and IIB were treated with 15-DS alone (4 mg/kg/day) for p.o. days 0-9. Groups IC and IIC were treated with cyclosporine A (10-40 mg/kg/day) for p.o. days 0-30. Groups ID and IID received a combination of 15-DS (for p.o. days 0-9) and CsA (for p.o. days 0-30). Groups IA and IIA served as control and received no medication. The mean graft survival was 11.0 days for group IA, 28.2 days for group IB (P less than 0.05; IB vs. IA), 32.4 days for group IC, and 43.1 days for group ID (P less than 0.025; ID vs. IA). After renal transplantation, the corresponding figures were 12.3 days for group IIA, 8.5 days for group IIB, 30.4 days for group IIC and 148.9 days for group IID (P less than 0.025; IID vs. IIA). After cardiac and renal transplantation, acute rejection was the main cause of graft failure. Treatment-related side effects, mainly gastrointestinal complications, were observed only in primates, who were treated with 15-DS alone. After cardiac transplantation, permanent graft non-reactivity was not achieved, but a delayed rejection occurred within a mean of 21.8 days after immunosuppression had been stopped. Following renal transplantation, graft nonreactivity was also not achieved in groups IIB and IIC. In group IID, however, 4 of 8 animals (50%) were graft-tolerant 340, 256, 244, and 164 days after treatment discontinuation. Thus, the combination of 15-DS and CsA led to a significant prolongation of graft survival in both groups. Long-term nonreactivity was achieved only after renal transplantation, when initially treated with 15-DS and CsA.

Use of manganese in RT-PCR eliminates PCR artifacts resulting from DNase I digestion.

The precise quantification of rare mRNA copies from intronless genes by reverse transcription polymerase chain reaction (RT-PCR) requires the complete removal of genomic DNA because discrimination of cDNA and DNA amplification products by differing sizes of PCR products is not possible. Elimination of DNA is achieved by treating the RNA sample with RNase-free DNase I before RT-PCR. The lack of a PCR product from DNase-treated RNA samples before RT is usually accepted as a proof of efficient DNA destruction. However, this may vary depending on the metal cofactor used in the DNase I cleavage. Treating DNA-contaminated RNA samples with DNase I and magnesium as a cofactor creates a negative PCR control after digestion without further RT. Paradoxically, after additional RT-PCR, the original intron-containing DNA fragment size may be produced again. In the presence of manganese as cofactor, RT-created DNA fragments do not appear. This is because in the presence of manganese, DNase I cleaves both DNA strands at approximately the same site, yielding DNA fragments that are blunt-ended or that have protruding termini of only one or two nucleotides in length. However, overlapping fragments with the potential to recombine are created by DNase digestion with magnesium as cofactor. Because one cannot differentiate between a PCR signal produced by RNA and one produced by recombined DNA after DNase I digestion and RT, all such DNase I assays should be performed with manganese instead of magnesium.