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Campylobacter jejuni remains one of the major gut pathogens of our time. Its zoonotic nature and wide-spread distribution in industrialized countries calls for a quick and reliable diagnostic tool. Antibody-based detection presents a suitable means to identify pathogenic bacteria. However, the knowledge about immunodominant targets is limited. Thus, an approach is presented, which allows for the rapid screening of numerous cDNA derived expression clones to identify novel antigens. The deeper understanding of immunodominant proteins assists in the design of diagnostic tools and furthers the insight into the bacterium's pathogenicity as well as revealing potential candidates for vaccination. We have successfully screened 1536 clones of an expression library to identify 22 proteins that have not been described as immunodominant before. After subcloning the corresponding 22 genes and expression of full-length proteins, we investigated the immunodominant character by microarrays and ELISA. Subsequently, seven proteins were selected for epitope mapping. For cj0669 and cj0920c linear epitopes were identified. For cj0669, specificity assays revealed a specific linear epitope site. Consequently, an eleven amino acid residue sequence TLIKELKRLGI was analyzed via alanine scan, which revealed the glycine residue to be significant for binding of the antibody. The innovative approach presented herein of generating cDNAs of prokaryotes in combination with a microarray platform rendering time-consuming purification steps obsolete has helped to illuminate novel immunodominant proteins of C.jejuni. The findings of a specific linear epitope pave the way for a plethora of future research and the potential use in diagnostic applications such as serological screenings. Moreover, the current approach is easily adaptable to other highly relevant bacteria making it a formidable tool for the future discovery of antigens and potential biomarkers. Consequently, it is desirable to simplify the identification of structural epitopes, as this would extend the spectrum of novel epitopes to be detected.

Background: PTEN hamartoma tumour syndrome (PHTS) encompasses several clinical syndromes with germline mutations in the PTEN tumour suppressor gene, including Cowden syndrome which is characterised by an increased risk of breast and thyroid cancers. Because PHTS is rare, data regarding cancer risks and genotype-phenotype correlations are limited. The objective of this study was to better define cancer risks in this syndrome with respect to the type and location of PTEN mutations., Methods: 154 PHTS individuals with a deleterious germline PTEN mutation were recruited from the activity of the Institut Bergonié genetic laboratory. Detailed phenotypic information was obtained for 146 of them. Age and sex adjusted standardised incidence ratio (SIR) calculations, cumulative cancer risk estimations, and genotype-phenotype analyses were performed., Results: Elevated SIRs were found mainly for female breast cancer (39.1, 95% CI 24.8 to 58.6), thyroid cancer in women (43.2, 95% CI 19.7 to 82.1) and in men (199.5, 95% CI 106.39 to 342.03), melanoma in women (28.3, 95% CI 7.6 to 35.4) and in men (39.4, 95% CI 10.6 to 100.9), and endometrial cancer (48.7, 95% CI 9.8 to 142.3). Cumulative cancer risks at age 70 were 85% (95% CI 70% to 95%) for any cancer, 77% (95% CI 59% to 91%) for female breast cancer, and 38% (95% CI 25% to 56%) for thyroid cancer. The risk of cancer was two times greater in women with PHTS than in men with PHTS (p<0.05)., Conclusions: This study shows a considerably high cumulative risk of cancer for patients with PHTS, mainly in women without clear genotype-phenotype correlation for this cancer risk. New recommendations for the management of PHTS patients are proposed.

Background: Bone-to-tendon healing after rotator cuff repairs is mainly impaired by poor tissue quality. The tenocytes of chronic rotator cuff tendon tears are not able to synthesize normal fibrocartilaginous extracellular matrix (ECM). We hypothesized that in the presence of platelet-released growth factors (PRGF), tenocytes from chronically retracted rotator cuff tendons proliferate and synthesize the appropriate ECM proteins., Materials and Methods: Tenocytes from 8 patients with chronic rotator cuff tears were cultured for 4 weeks in 2 different media: standard medium (Iscove's Modified Dulbecco's Media + 10% fetal calf serum + 1% nonessential amino acids + 0.5 μg/mL ascorbic acid) and media with an additional 10% PRGF. Cell proliferation was assessed at 7, 14, 21, and 28 days. Messenger (m)RNA levels of collagens I, II, and X, decorin, biglycan, and aggrecan were analyzed using real time reverse-transcription polymerase chain reaction. Immunocytochemistry was also performed., Results: The proliferation rate of tenocytes was significantly higher at all time points when cultured with PRGF. At 21 days, the mRNA levels for collagens I, II, and X, decorin, aggrecan, and biglycan were significantly higher in the PRGF group. The mRNA data were confirmed at protein level by immunocytochemistry., Conclusions: PRGFs enhance tenocyte proliferation in vitro and promote synthesis of ECM to levels similar to those found with insertion of the normal human rotator cuffs., Clinical Relevance: Biologic augmentation of repaired rotator cuffs with PRGF may enhance the properties of the repair tissue. However, further studies are needed to determine if application of PRGF remains safe and effective in long-term clinical studies., Level of Evidence: Basic Science Study, Cell Biology., (Copyright © 2013 Journal of Shoulder and Elbow Surgery Board of Trustees. Published by Mosby, Inc. All rights reserved.)

Background: Meropenem is an effective β-lactam antibiotic that is frequently used to treat serious infections in both intensive care unit (ICU) and febrile neutropenic hematology/oncology (Hem/Onc) patients. Studies suggest that to be effective, meropenem concentrations must be maintained above the inhibitory concentrations for the majority of a dosing interval. However, the pharmacokinetics (PK) of meropenem seem to differ in critically ill patients compared with healthy or less ill subjects used to select labeled dosing regimens., Objectives: This study was designed to investigate meropenem PK in critically ill patients and to see how often standard dosing regimens produced adequate plasma concentrations. A secondary objective was to investigate how achieved concentrations were related to outcomes (morbidity and mortality) in these patients., Methods: Meropenem plasma concentrations over time were measured using a high pressure liquid chromatography assay in febrile Hem/Onc and ICU patients who were treated with standard meropenem dosing schedules. Outcomes such as fever control and survival were assessed in these patients and compared with individual meropenem PK data and with recommended target concentrations., Results: A total of 25 subjects including 10 febrile Hem/Onc and 15 ICU patients with a variety of serious illnesses and baseline renal function were studied. Mean peak concentrations were less variable than were pre-dose concentrations. Post peak and trough concentrations were often below recommended minimum inhibitory concentrations. Both clearance and volumes of distribution were greater than reported in less ill subjects, only in part explained by increased renal clearance. Therefore, serum concentrations often did not exceed recommended concentration targets even for moderately sensitive organisms. Inadequate concentrations were especially common in the mostly ill, febrile neutropenic Hem/Onc subjects and seemed to explain at least some therapeutic failures. Conversely, drug accumulation occurred in ICU subjects with decreased renal function., Conclusions: Standard meropenem dosing regimens were inadequate in many critically ill febrile, neutropenic Hem/Onc, and septic ICU patients. These data suggest a role for meropenem concentration monitoring in such patients.

Objective: Depression and anxiety lead to increased morbidity and mortality in patients with coronary artery disease (CAD). Medication nonadherence is one possible pathway contributing to adverse outcome, but it is unknown how either depression or anxiety itself influences adherence compared to combined depressive-anxious comorbidity. The aim of the study was to evaluate the influence of simultaneous depressive and anxious symptoms on medication adherence in patients with stable CAD., Methods: Between 02/2009 and 06/2010 we examined the association between current depressive and anxious symptoms with medication nonadherence in a cross-sectional study of 606 inpatients with stable CAD. Symptoms were assessed by using the Hospital Anxiety and Depression Scale. Morisky Medical Adherence Scale measured medication adherence., Results: Depressive and anxious symptoms were weakly and independently associated with medication nonadherence (r=0.28, p<0.01 and r=0.27, p<0.01 respectively). Compared to non-depressed, patients with depressive symptoms had an up to 3.6-fold odds, those with anxious symptoms an up to 3.2-fold odds of nonadherence. The presence of combined anxiety and depressive symptoms was also weakly correlated with adherence (r=0.30, p<0.01). The risk for nonadherence in patients suffering from both anxiety and depression was up to 4.4 times higher compared to patients without symptoms., Conclusion: Apart from depressive symptoms, anxiety is a second important and independent marker for nonadherence in patients with coronary artery disease. The negative effect of anxiety on medication adherence increases in case of comorbid depressive symptoms. Future studies addressing medication adherence should focus more on anxious-depressive comorbidity than on singular depressive or anxious symptoms., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Background: Hospital-based cancer registries (HBR) of Comprehensive cancer centers (CCC) named "Enquête permanente cancer (EPC)" could represent an important information source of data for a population-based cancer registry (PBR). The main purpose of this study was to evaluate the concordance between data from HBR of the CCC for regions of Bordeaux and South-Western France and data from PBR of the General cancer registry of Gironde and the completeness for the year 2005, in order to explore the possibility for immediate integration of data collected by the HBR in the database of the PBR and to reduce the systematic consultation of medical records for cancer cases notified by the HBR., Methods: We estimated the proportion of concordant cases between the HBR and the PBR and the unweighted and weighted Cohen's Kappa coefficient for certain variables., Results: Among 1151 cases notified by the HBR, 1095 (positive predictive value of HBR: 95%) were included in the PBR database. For these admissible cases, sex, birth date and place for residence postal code at diagnosis were concordant in more than 94%. Concordance of cancer incidence date was 50% but difference between two dates was lower than 1 month in 91% of cases. Concordance of cancer topography and morphology was respectively 72% and 78%. Cancer progression, estimated by TNM classification was concordant in 96% of cases. The Kappa coefficient was 0.95 for pT, and respectively 0.92 for pN and M., Conclusion: Automatic data integration from the HBR database to the PBR database without consulting medical record will be possible in Gironde. But, topography and morphology will be systematically recoded using pathologist records. And, if cancer progression (coded by TNM classification) is missing in the HBR database, medical records will be consulted in order to complete this data in the PBR database., (Copyright © 2012 Elsevier Masson SAS. All rights reserved.)

Background: The regenerative capacity of the liver is an essential pre-condition for the successful application of partial hepatectomy. However, the actual kinetics of functional recovery remains unspecified and no adequate tool for its clinical monitoring has yet been available., Methods: Eighty-five patients receiving major hepatectomy were investigated from the preoperative evaluation until 12 weeks after surgery. Liver function was determined by the LiMAx test for the enzymatic capacity of cytochrome P450 1A2. Liver volume was determined by volumetric analysis of repeated computer tomography scans. Functional and volume recovery were compared during follow-up., Results: Major hepatectomy decreased liver function capacity to 35.7 ± 13.8% of preoperative function. It was shown that functional recovery already reaches 77.2 ± 33.5% of preoperative values within 10 days. The actual kinetics were dependent from the type and extent of hepatectomy. Complete functional restoration was achieved within 12 weeks, while liver volume still remained at 73.2 ± 14.8% of preoperative. A constant but interindividually variable correlation between function and volume was observed at all points in time., Conclusion: Partial hepatectomy leads to fast and complete functional recovery, while volume recovery is delayed and remains often incomplete. The functional recovery is mainly influenced by the preoperative liver function, the residual liver volume, and by obesity.

Background and Purpose: Computer navigation in total knee arthroplasty is somewhat controversial. We have previously shown that femoral component positioning is more accurate with computed navigation than with conventional implantation techniques, but the clinical impact of this is unknown. We now report the 5-year outcome of our previously reported 2-year outcome study., Methods: 78 of initially 84 patients (80 of 86 knees) were clinically and radiographically reassessed 5 (5.1-5.9) years after conventional, image-based, and image-free total knee arthroplasty. The methodology was identical to that used preoperatively and at 2 years, including the Knee Society score (KSS) and the functional score (FS), and AP and true lateral standard radiographs., Results: Although a more accurate femoral component positioning in the navigated groups was obtained, clinical outcome, number of reoperations, KSS, FS, and range of motion were similar between the groups., Interpretation: The increased costs and time for navigated techniques did not translate into better functional and subjective medium-term outcome compared to conventional techniques.

Objectives: To demonstrate the potential benefits of biochemical axial T2 mapping of intervertebral discs (IVDs) regarding the detection and grading of early stages of degenerative disc disease using 1.5-Tesla magnetic resonance imaging (MRI) in a clinical setting., Methods: Ninety-three patients suffering from lumbar spine problems were examined using standard MRI protocols including an axial T2 mapping protocol. All discs were classified morphologically and grouped as "healthy" or "abnormal". Differences between groups were analysed regarding to the specific T2 pattern at different regions of interest (ROIs)., Results: Healthy intervertebral discs revealed a distinct cross-sectional T2 value profile: T2 values were significantly lower in the annulus fibrosus compared with the nucleus pulposus (P = 0.01). In abnormal IVDs, T2 values were significantly lower, especially towards the centre of the disc representing the expected decreased water content of the nucleus (P = 0.01). In herniated discs, ROIs within the nucleus pulposus and ROIs covering the annulus fibrosus showed decreased T2 values., Conclusions: Axial T2 mapping is effective to detect early stages of degenerative disc disease. There is a potential benefit of axial T2 mapping as a diagnostic tool, allowing the quantitative assessment of intervertebral disc degeneration.

Background: Detection of immunogenic proteins remains an important task for life sciences as it nourishes the understanding of pathogenicity, illuminates new potential vaccine candidates and broadens the spectrum of biomarkers applicable in diagnostic tools. Traditionally, immunoscreenings of expression libraries via polyclonal sera on nitrocellulose membranes or screenings of whole proteome lysates in 2-D gel electrophoresis are performed. However, these methods feature some rather inconvenient disadvantages. Screening of expression libraries to expose novel antigens from bacteria often lead to an abundance of false positive signals owing to the high cross reactivity of polyclonal antibodies towards the proteins of the expression host. A method is presented that overcomes many disadvantages of the old procedures., Results: Four proteins that have previously been described as immunogenic have successfully been assessed immunogenic abilities with our method. One protein with no known immunogenic behaviour before suggested potential immunogenicity.We incorporated a fusion tag prior to our genes of interest and attached the expressed fusion proteins covalently on microarrays. This enhances the specific binding of the proteins compared to nitrocellulose. Thus, it helps to reduce the number of false positives significantly. It enables us to screen for immunogenic proteins in a shorter time, with more samples and statistical reliability. We validated our method by employing several known genes from Campylobacter jejuni NCTC 11168., Conclusions: The method presented offers a new approach for screening of bacterial expression libraries to illuminate novel proteins with immunogenic features. It could provide a powerful and attractive alternative to existing methods and help to detect and identify vaccine candidates, biomarkers and potential virulence-associated factors with immunogenic behaviour furthering the knowledge of virulence and pathogenicity of studied bacteria.

Objective: Developing an efficient tool for accurate three-dimensional imaging from projections measured with C-arm systems., Material and Methods: A circle-plus-arc trajectory, which is complete and thus amenable to accurate reconstruction, is used. This trajectory is particularly attractive as its implementation does not require moving the patient. For reconstruction, we use the "M-line method", which allows processing the data in the efficient filtered backprojection mode. This method also offers the advantage of not requiring an ideal data acquisition geometry, i.e., the M-line algorithm can account for known deviations in the scanning geometry, which is important given that sizeable deviations are generally encountered in C-arm imaging., Results: A robust implementation scheme of the "M-line method" that applies straightforwardly to real C-arm data is presented. In particular, a numerically stable technique to compute the view-dependent derivative with respect to the source trajectory parameter is applied, and an efficient way to compute the π-line backprojection intervals via a polygonal weighting mask is presented. Projection data of an anthropomorphic thorax phantom were acquired on a medical C-arm scanner and used to demonstrate the benefit of using a complete data acquisition geometry with an accurate reconstruction algorithm versus using a state-of-the-art implementation of the conventional Feldkamp algorithm with a circular short scan of cone-beam data. A significant image quality improvement based on visual assessment is shown in terms of cone-beam artifacts.

All positive strand RNA viruses are known to replicate their genomes in close association with intracellular membranes. In case of the hepatitis C virus (HCV), a member of the family Flaviviridae, infected cells contain accumulations of vesicles forming a membranous web (MW) that is thought to be the site of viral RNA replication. However, little is known about the biogenesis and three-dimensional structure of the MW. In this study we used a combination of immunofluorescence- and electron microscopy (EM)-based methods to analyze the membranous structures induced by HCV in infected cells. We found that the MW is derived primarily from the endoplasmic reticulum (ER) and contains markers of rough ER as well as markers of early and late endosomes, COP vesicles, mitochondria and lipid droplets (LDs). The main constituents of the MW are single and double membrane vesicles (DMVs). The latter predominate and the kinetic of their appearance correlates with kinetics of viral RNA replication. DMVs are induced primarily by NS5A whereas NS4B induces single membrane vesicles arguing that MW formation requires the concerted action of several HCV replicase proteins. Three-dimensional reconstructions identify DMVs as protrusions from the ER membrane into the cytosol, frequently connected to the ER membrane via a neck-like structure. In addition, late in infection multi-membrane vesicles become evident, presumably as a result of a stress-induced reaction. Thus, the morphology of the membranous rearrangements induced in HCV-infected cells resemble those of the unrelated picorna-, corona- and arteriviruses, but are clearly distinct from those of the closely related flaviviruses. These results reveal unexpected similarities between HCV and distantly related positive-strand RNA viruses presumably reflecting similarities in cellular pathways exploited by these viruses to establish their membranous replication factories.

Objective: To determine whether administration of lidocaine during ischemia and reperfusion in horses results in concentrations in smooth muscle sufficient to protect against the negative consequences of ischemia-reperfusion injury on smooth muscle motility., Animals: 12 horses., Procedures: Artificial ischemia and reperfusion injury of jejunal segments was induced in vivo in conjunction with lidocaine treatment during ischemia (IRL) or without lidocaine treatment (IR). Isometric force performance was measured in vitro in IRL and IR smooth muscle preparations with and without additional in vitro application of lidocaine. Lidocaine concentrations in smooth muscle were determined by means of high-performance liquid chromatography. To assess the influence of lidocaine on membrane permeability, activity of creatine kinase and lactate dehydrogenase released by in vitro incubated tissues was determined biochemically., Results: In vivo administration of lidocaine allowed maintenance of contractile performance after an ischemia and reperfusion injury. Basic contractility and frequency of contractions were significantly increased in IRL smooth muscle tissues in vitro. Additionally, in vitro application of lidocaine achieved further improvement of contractility of IR and IRL preparations. Only in vitro application of lidocaine was able to ameliorate membrane permeability in smooth muscle of IR and IRL preparations. Lidocaine accumulation could be measured in in vivo treated samples and serum., Conclusions and Clinical Relevance: In vivo lidocaine administration during ischemia and reperfusion had beneficial effects on smooth muscle motility. Initiating lidocaine treatment during surgery to treat colic in horses may improve lidocaine's prokinetic features by protecting smooth muscle from effects of ischemia and reperfusion injury.

Current water quality criteria (WQC) regulations on copper toxicity to biota are still based on total dissolved (<0.4 μm membrane filter) copper concentrations with a hardness modification for freshwaters. There are however ongoing efforts to incorporate metal speciation in WQC and toxicity regulations (such as the biotic ligand model-BLM) for copper and other metals. Here, we show that copper accumulation and growth inhibition of the Baltic macroalga Ceramium tenuicorne exposed to copper in artificial seawater at typical coastal and estuarine DOC concentrations (similar to 2-4 mg/L-C as fulvic acid) are better correlated to weakly complexed and total dissolved copper concentrations rather than the free copper concentration [Cu2+]. Our results using a combination of competitive ligand exchange-adsorptive cathodic stripping voltammetry (CLE-ACSV) measurements and model calculations (using visual MINTEQ incorporating the Stockholm Humic Model) show that copper accumulation in C. tenuicorne only correlates linearly well to [Cu2+] at relatively high [Cu2+] and in the absence of fulvic acid. Thus the FIAM fails to describe copper accumulation in C. tenuicorne at copper and DOC concentrations typical of most marine waters. These results seem to indicate that at ambient total dissolved copper concentration in coastal and estuarine waters, C. tenuicorne might be able to access a sizable fraction of organically complexed copper when free copper concentration to the cell membrane is diffusion limited.

Stem cell based autologous grafting has recently gained mayor interest in various surgical fields for the treatment of extensive tissue defects. CD34(+) and CD133(+) cells that can be isolated from the pool of bone marrow mononuclear cells (BMC) are capable of differentiating into mature endothelial cells in vivo. These endothelial progenitor cells (EPC) are believed to represent a major portion of the angiogenic regenerative cells that are released from bone marrow when tissue injury has occurred. In recent years tissue engineers increasingly looked at the process of vessel neoformation because of its major importance for successful cell grafting to replace damaged tissue. Up to now one of the greatest problems preventing a clinical application is the large scale of expansion that is required for such purpose. We established a method to effectively enhance the expansion of CD34(+) and CD133(+) cells by the use of platelet-released growth factors (PRGF) as a media supplement. PRGF were prepared from thrombocyte concentrates and used as a media supplement to iscove's modified dulbecco's media (IMDM). EPC were immunomagnetically separated from human bone morrow monocyte cells and cultured in IMDM + 10% fetal calf serum (FCS), IMDM + 5%, FCS + 5% PRGF and IMDM + 10% PRGF. We clearly demonstrate a statistically significant higher and faster cell proliferation rate at 7, 14, 21, and 28 days of culture when both PRGF and FCS were added to the medium as opposed to 10% FCS or 10% PRGF alone. The addition of 10% PRGF to IMDM in the absence of FCS leads to a growth arrest from day 14 on. In histochemical, immunocytochemical, and gene-expression analysis we showed that angiogenic and precursor markers of CD34(+) and CD133(+) cells are maintained during long-term culture. In summary, we established a protocol to boost the expansion of CD34(+) and CD133(+) cells. Thereby we provide a technical step towards the clinical application of autologous stem cell transplantation.

VASP is an actin-regulatory protein that links signalling to remodelling of the cytoskeleton. We investigated the role of VASP during entry of herpes simplex viruses into epithelial MDCKII cells. As VASP functions are regulated by phosphorylations, the phosphorylation pattern was determined upon infection. Phosphorylated VASP decreased temporarily at 15 and 30 min after infection. The impact of phosphorylated VASP was addressed by overexpression of phosphomimetic VASP mutants. Our results revealed that phosphorylated VASP slightly reduced the number of infected cells. Expression studies with deletion mutants further indicated minor effects of VASP on infection efficiency, whereas RNA interference studies demonstrated that reduced VASP expression did not suppress infection. We conclude that VASP activities alone may contribute to herpes simplex virus infection to only a minor extent.

A total of 3,050 German Angus (Aberdeen Angus x German dual-purpose breeds), Charolais, Hereford, Limousin, and German Simmental calves were used to examine temperament traits of beef cattle using 2 different test procedures. The chute test and the flight-speed test have been validated in terms of routine on-farm applicability. Behavior tests were performed in 2006 and 2007 on 24 commercial beef cattle farms located in the northern and eastern part of Germany. A single, trained observer assigned subjective scores to characterize the behavior of each animal during restraint in the head gate (calm, restless shifting, squirming, vigorous movement, violent struggling) and when leaving the chute (walk, trot, run, jumping out of the chute). Breed was a significant source of variation in chute scores and flight-speed scores (P < 0.001). Charolais and Limousin cattle had the greatest scores in both traits, whereas Herefords had the least (P < 0.001) chute scores. German Angus and Hereford calves had the least (P < 0.001) flight-speeds, indicating that these breeds have a more favorable temperament. Temperament scores differed significantly between male and female calves (P < 0.01), with females scoring better for both traits. Average daily BW gains of the calves were significantly influenced by effects of breed (P < 0.001) and sex (P < 0.001) of the calves. Heritabilities were estimated for chute- and flight-speed scores of beef cattle. They were least for chute score and flight-speed score of Limousin cattle with values of 0.11. In contrast, greatest heritabilities were 0.33 for chute score and 0.36 for flight-speed score of Hereford cattle. Genetic correlations were estimated among both temperament traits, with values between 0.57 and 0.98. Chute scores and visual flight-speed scores were negatively correlated with daily BW gain of the calves in most breeds. The results presented in this paper indicate that on-farm evaluation of beef cattle temperament is possible, either using the chute test or the flight-speed test. Genetic selection seems to be promising to improve temperament traits of beef cattle without decreasing production traits like ADG of the calves.

Reasons for Performing Study: Post operative ileus (POI) in horses is a severe complication after colic surgery. A commonly used prokinetic drug is lidocaine, which has been shown to have stimulatory effects on intestinal motility. The cellular mechanisms through which lidocaine affects smooth muscle activity are not yet known., Objectives: To examine the effects of lidocaine on smooth muscle in vitro and identify mechanisms by which it may affect the contractility of intestinal smooth muscle., Hypothesis: Ischaemia and reperfusion associated with intestinal strangulation can cause smooth muscle injury. Consequently, muscle cell functionality and contractile performance is decreased. Lidocaine can improve basic cell functions and thereby muscle cell contractility especially in ischaemia-reperfusion-challenged smooth muscle., Methods: To examine the effects of lidocaine on smooth muscle function directly, isometric force performance was measured in vitro in noninjured and in vivo ischaemia-reperfusion injured smooth muscle tissues. Dose-dependent response of lidocaine was measured in both samples. To assess membrane permeability as a marker of basic cell function, release of creatine kinase (CK) was measured by in vitro incubations., Results: Lidocaine-stimulated contractility of ischaemia-reperfusion injured smooth muscle was more pronounced than that of noninjured smooth muscle. A 3-phasic dose-dependency was observed with an initial recovery of contractility especially in ischaemia-reperfusion injured smooth muscle followed by a plateau phase where contractility was maintained over a broad concentration range. CK release was decreased by lidocaine., Conclusion: Lidocaine may improve smooth muscle contractility and basic cell function by cellular repair mechanisms which are still unknown. Improving contractility of smooth muscle after ischaemia-reperfusion injury is essential in recovery of propulsive intestinal motility., Potential Relevance: Characterisation of the cellular mechanisms of effects of lidocaine, especially on ischaemia-reperfusion injured smooth muscle, may lead to improved treatment strategies for horses with POI.

Clinical assessments after Total Knee Arthroplasty (TKA) show persisting pain after implantation in over 20% of patients. Impingement of soft tissue around the knee, due to imprecise geometry of the tibial implant, can be one reason for persisting ailment. Two hundred and thirty seven MRI scans were evaluated using an active contour detection algorithm (snake) to obtain a high-resolution mean anatomical shape of the tibial plateau. Differences between female and male, older and younger (40) and left and right averaged shapes were determined. The shapes obtained were asymmetric throughout. Absolute differences between the subgroups fell short of inter-individual variations represented by calculated one-sigma confidence intervals. Our results indicate that a differentiation in TKA tibial plateau design by gender, age, or side is of minor relevance.

Starburst galaxies exhibit in their central regions a highly increased rate of supernovae, the remnants of which are thought to accelerate energetic cosmic rays up to energies of approximately 10(15) electron volts. We report the detection of gamma rays--tracers of such cosmic rays--from the starburst galaxy NGC 253 using the High Energy Stereoscopic System (H.E.S.S.) array of imaging atmospheric Cherenkov telescopes. The gamma-ray flux above 220 billion electron volts is F = (5.5 +/- 1.0(stat) +/- 2.8(sys)) x 10(-13) cm(-2) s(-1), implying a cosmic-ray density about three orders of magnitude larger than that in the center of the Milky Way. The fraction of cosmic-ray energy channeled into gamma rays in this starburst environment is five times as large as that in our Galaxy.

AMP-activated protein kinase (AMPK) senses changes in the intracellular AMP/ATP ratio, switching off energy-consuming processes and switching on catabolic pathways in response to energy depletion. Here, we show that AMPK down-regulates rRNA synthesis under glucose restriction by phosphorylating the RNA polymerase I (Pol I)-associated transcription factor TIF-IA at a single serine residue (Ser-635). Phosphorylation by AMPK impairs the interaction of TIF-IA with the TBP-containing promoter selectivity factor SL1, thereby precluding the assembly of functional transcription initiation complexes. Mutation of Ser-635 compromises down-regulation of Pol I transcription in response to low energy supply, supporting that activation of AMPK adapts rRNA synthesis to nutrient availability and the cellular energy status.

The accretion of matter onto a massive black hole is believed to feed the relativistic plasma jets found in many active galactic nuclei (AGN). Although some AGN accelerate particles to energies exceeding 10(12) electron volts and are bright sources of very-high-energy (VHE) gamma-ray emission, it is not yet known where the VHE emission originates. Here we report on radio and VHE observations of the radio galaxy Messier 87, revealing a period of extremely strong VHE gamma-ray flares accompanied by a strong increase of the radio flux from its nucleus. These results imply that charged particles are accelerated to very high energies in the immediate vicinity of the black hole.

The very large collection area of ground-based gamma-ray telescopes gives them a substantial advantage over balloon or satellite based instruments in the detection of very-high-energy (>600 GeV) cosmic-ray electrons. Here we present the electron spectrum derived from data taken with the High Energy Stereoscopic System (H.E.S.S.) of imaging atmospheric Cherenkov telescopes. In this measurement, the first of this type, we are able to extend the measurement of the electron spectrum beyond the range accessible to direct measurements. We find evidence for a substantial steepening in the energy spectrum above 600 GeV compared to lower energies.

State-of-the-art filtered backprojection (FBP) algorithms often define the filtering operation to be performed along oblique filtering lines in the detector. A limited scan field of view leads to the truncation of those filtering lines, which causes artifacts in the final reconstructed volume. In contrast to the case where filtering is performed solely along the detector rows, no methods are available for the case of oblique filtering lines. In this work, the authors present two novel truncation correction methods which effectively handle data truncation in this case. Method 1 (basic approach) handles data truncation in two successive preprocessing steps by applying a hybrid data extrapolation method, which is a combination of a water cylinder extrapolation and a Gaussian extrapolation. It is independent of any specific reconstruction algorithm. Method 2 (kink approach) uses similar concepts for data extrapolation as the basic approach but needs to be integrated into the reconstruction algorithm. Experiments are presented from simulated data of the FORBILD head phantom, acquired along a partial-circle-plus-arc trajectory. The theoretically exact M-line algorithm is used for reconstruction. Although the discussion is focused on theoretically exact algorithms, the proposed truncation correction methods can be applied to any FBP algorithm that exposes oblique filtering lines.

In the past few decades, several models have predicted an energy dependence of the speed of light in the context of quantum gravity. For cosmological sources such as active galaxies, this minuscule effect can add up to measurable photon-energy dependent time lags. In this Letter a search for such time lags during the High Energy Stereoscopic System observations of the exceptional very high energy flare of the active galaxy PKS 2155-304 on 28 July 2006 is presented. Since no significant time lag is found, lower limits on the energy scale of speed of light modifications are derived.

Background: Functional inactivation of the tumor suppressor Smad4 in colorectal and pancreatic carcinogenesis occurs coincident with the transition to invasive growth. Breaking the basement membrane (BM) barrier, a prerequisite for invasive growth, can be due to tumor induced proteolytic tissue remodeling or to reduced synthesis of BM molecules by incipient tumor cells. Laminin-332 (laminin-5), a heterotrimeric BM component composed of alpha 3-, beta 3- and gamma 2-chains, has recently been identified as a target structure of Smad4 and represents the first example for expression control of an essential BM component by a tumor and invasion suppressor. Biochemically Smad4 is a transmitter of signals of the TGFbeta superfamily of cytokines. We have reported previously, that Smad4 functions as a positive transcriptional regulator of constitutive and of TGFbeta-induced transcription of all three genes encoding Laminin-332, LAMA3, LAMB3 and LAMC2., Methods: Promoter-reporter constructs harboring 4 kb upstream regions, each of the three genes encoding Laminin-322 as well as deletion and mutations constructs were established. Promoter activities and TGFbeta induction were assayed through transient transfections in Smad4-negative human cancer cells and their stable Smad4-positive derivatives. Functionally relevant binding sites were subsequently confirmed through chromatin immunoprecipitation., Results: Herein, we report that Smad4 mediates transcriptional regulation through three different mechanisms, namely through Smad4 binding to a functional SBE site exclusively in the LAMA3 promoter, Smad4 binding to AP1 (and Sp1) sites presumably via interaction with AP1 family components and lastly a Smad4 impact on transcription of AP1 factors. Whereas Smad4 is essential for positive regulation of all three genes, the molecular mechanisms are significantly divergent between the LAMA3 promoter as compared to the LAMB3 and LAMC2 promoters., Conclusion: We hypothesize that this divergence in modular regulation of the three promoters may lay the ground for uncoupled regulation of Laminin-332 in Smad4-deficient tumor cells in response to stromally expressed cytokines acting on budding tumor cells.

In this paper, a novel geometric calibration method for C-arm cone-beam scanners is presented which allows the calibration of the circle-plus-arc trajectory. The main idea is the separation of the trajectory into two circular segments (circle segment and arc segment) which are calibrated independently. This separation makes it possible to reuse a calibration phantom which has been successfully applied in clinical environments to calibrate numerous routinely used C-arm systems. For each trajectory segment, the phantom is placed in an optimal position. The two calibration results are then combined by computing the transformation the phantom underwent between the independent calibration runs. This combination can be done in a post-processing step by using standard linear algebra. The method is not limited to circle-plus-arc trajectories and works for any calibration procedure in which the phantom has a preferred orientation with respect to a trajectory segment. Results are presented for both simulated as well as real data acquired with a C-arm system. We also present the first image reconstruction results for the circle-plus-arc trajectory using real C-arm data.

Background: Smad4 is a tumour suppressor frequently inactivated in pancreatic and colorectal cancers. We have recently reported loss of Smad4 in every fourth carcinoma of the uterine cervix. Smad4 transmits signals from the TGF-beta superfamily of cytokines and functions as a versatile transcriptional co-modulator. The prevailing view suggests that the tumour suppressor function of Smad4 primarily resides in its capability to mediate TGF-beta growth inhibitory responses. However, accumulating evidence indicates, that the acquisition of TGF-beta resistance and loss of Smad4 may be independent events in the carcinogenic process. Through inducible reexpression of Smad4 in cervical cancer cells we wished to shed more light on this issue and to identify target genes implicated in Smad4 dependent tumor suppression., Methods: Smad4-deficient human C4-II cervical carcinoma cells were used to establish inducible Smad4 reexpression using the commercial Tet-ontrade mark system (Clontech). The impact of Smad4 reexpression on cell growth was analysed in vitro and in vivo. Transcriptional responses were assessed through profiling on cDNA macroarrays (Clontech) and validated through Northern blotting., Results: Clones were obtained that express Smad4 at widely varying levels from approximately physiological to 50-fold overexpression. Smad4-mediated tumour suppression in vivo was apparent at physiological expression levels as well as in Smad4 overexpressing clones. Smad4 reexpression in a dose-dependent manner was associated with transcriptional induction of the extracellular matrix-associated genes, BigH3, fibronectin and PAI-1, in response to TGF-beta. Smad4-dependent regulation of these secreted Smad4 targets is not restricted to cervical carcinoma cells and was confirmed in pancreatic carcinoma cells reexpressing Smad4 after retroviral transduction and in a stable Smad4 knockdown model. On the other hand, the classical cell cycle-associated TGF-beta target genes, c-myc, p21 and p15, remained unaltered., Conclusion: Our results show that Smad4-mediated tumour suppression in cervical cancer cells is not due to restoration of TGF-beta growth inhibitory responses. Rather, tumour cell-ECM interactions may be more relevant for Smad4-mediated tumour suppression. C4-II cells with a high level inducible Smad4 expression may serve as a model to indicate further Smad4 targets responsive to diverse environmental stimuli operative in vivo.

In computed tomography, analytical fan-beam (FB) and cone-beam (CB) image reconstruction often involves a view-dependent data differentiation. The implementation of this differentiation step is critical in terms of resolution and image quality. In this work, we present a new differentiation scheme that is robust to changes in the data acquisition geometry and to coarse view sampling. Our scheme was compared to two previously suggested methods, which we call the direct scheme and the chain-rule scheme. Image reconstructions were performed from computer-simulated data of the Shepp-Logan phantom, the FORBILD thorax phantom and a modified FORBILD head phantom. For FB reconstruction, we investigated three acquisition geometries: a circular, an ellipse-shaped and a square-shaped trajectory. For CB reconstruction, the circle-plus-line trajectory was considered. Image comparison showed that the new scheme performs consistently well when varying the scenario, in both FB and CB geometry, unlike the other two schemes.

The tumor suppressor Smad4 is involved in carcinogenesis mainly of the pancreas and colon. Functional inactivation of Smad4 is a genetically late event that occurs upon transition from premalignant stages to invasive and metastatic growth. Smad4 encodes an intracellular messenger common to all signalling cascades induced by members of the transforming growth factor-beta (TGF-beta) superfamily of cytokines. Despite extensive knowledge about the mechanisms of TGF-beta/Smad signal transduction, little is known about Smad4 targets involved in the transition to malignancy. The hallmark of invasive growth is a breakdown of the basement membrane (BM), a specialized sheet of extracellular matrix produced through cooperation of epithelial and stromal cells. Laminin-5, a heterotrimeric epithelial-derived BM component, is commonly lost in carcinomas but not in premalignant tumors. Herein, we report that in human colon and pancreatic tumor cells, Smad4 functions as a positive transcriptional regulator of all three genes encoding laminin-5. Coordinate re-expression of the three laminin-5 chains induced by reconstitution of Smad4 leads to secretion and deposition of the heterotrimeric molecule in BM-like structures. These data define the expression control of an essential BM component as a novel function for the tumor suppressor Smad4.

The detection of fast variations of the tera-electron volt (TeV) (10(12) eV) gamma-ray flux, on time scales of days, from the nearby radio galaxy M87 is reported. These variations are about 10 times as fast as those observed in any other wave band and imply a very compact emission region with a dimension similar to the Schwarzschild radius of the central black hole. We thus can exclude several other sites and processes of the gamma-ray production. The observations confirm that TeV gamma rays are emitted by extragalactic sources other than blazars, where jets are not relativistically beamed toward the observer.

The aim of this study was to understand how molecular determinants of epithelial cells influence initial infection by herpes simplex virus type 1 (HSV-1). Upon infection of the epithelial MDCKII cell line, enhanced association of virus particles with cells forming actin protrusions was observed, suggesting a putative role of actin dynamics in HSV-1 infection. Thus, the impact of the small Rho-like GTPases Rac1, Cdc42 and RhoA acting as key regulators of actin dynamics was addressed. Endogenous Rac1 and Cdc42 were temporarily activated at 15 and 30 min after HSV-1 infection. When constitutively active Cdc42 or Rac1 mutants were expressed transiently, a significant decrease in infectivity was observed, whereas expression of RhoA mutants had no influence. Furthermore, dominant-negative Cdc42 led to decreased infectivity, whereas dominant-negative Rac1 had no effect. So far, the study of potential effectors indicated that Rac1/Cdc42 mutants inhibited infectivity independently of p21-activated kinase (Pak1). The inhibitory effect of Rac1/Cdc42 mutant expression on HSV-1 infection was characterized further and it was found that binding, internalization and transport of HSV-1 were not affected by expression of Rac1/Cdc42 mutants. Thus, these results provide the first evidence for a role of Rac1/Cdc42 signalling during early HSV-1 infection and suggest a mechanism relying on virus-induced regulation of Rac1/Cdc42 activities.

Objective: We investigated the agreement of multislice computed tomography angiography (CTA) and magnetic resonance angiography (MRA) in the quantitative measurement of carotid artery stenosis. The dependency of the agreement of the chosen postprocessing procedures was also investigated., Methods: Fifty consecutive symptomatic patients were included in this study. In all patients, a CTA was performed with a 16-slice CT scanner. Within 30 days, the extracranial vessels were examined using a combined time-of-flight and contrast-enhanced MRA. The CT data sets were used to calculate the degree of stenosis according to the North American Symptomatic Carotid Endarterectomy Trial, European Carotid Surgery Trial, and Common Carotid methods by means of the 1-mm thick, transverse raw data (RD), a sagittal maximum-intensity projection reconstruction, and sagittal multiplanar reconstruction. In addition, a semiautomated analysis was done using a specialized postprocessing software. For all combinations of postprocessing procedures and methods of calculating the degree of stenosis, the correlation coefficient and the agreement based on Bland/Altman plots were calculated., Results: Eleven of the 100 primarily included carotid arteries could not be evaluated. The correlation coefficients for all combinations were comparable and lied in the interval between 0.932 and 0.787. The best correlation was found for the combination of RD/sagittal multiplanar reconstruction and ECST method. The evaluation of the agreement gave a systematic overestimation of CTA between 1.9% and 10.7% with a 95% confidence interval between +/-26.7% and +/-43.3%. With the semiautomated postprocessing software, additional 33 vessels could not be evaluated. The agreement of the calculated degrees of stenoses was worse than that of the planar procedures., Conclusions: CTA and MRA had a feasible agreement in measuring the degree of stenosis of the carotid arteries. The best result could be obtained for the evaluation of the RD and the NASCET method. In this case one has to take into account a systematic overestimation of CTA of 1.9%. The combination with an additional reconstructive postprocessing procedure did not improve the result but might be useful for the radiologist to identify the location of the closest narrowing.

Introduction: The treatment of unstable cervical spine injuries with the halo vest represents an established procedure. So far no data reflecting the quality of life of patients following a halo vest treatment are available. Elderly people make up a large part of the inpatients in our hospital. Therefore special attention is payed to this group of patients in this study., Methods: In this study 41 patients (average age of 51.8 +/- 23.5 years) with an unstable injury of the upper cervical spine were investigated. All of them underwent a halo vest therapy in our hospital during 1988-2003. The health-related quality of life was assessed in the mean 8.0 years after the trauma by using the SF-36 Health Survey. Additionally, the incidence of complications and the union rate over time were observed., Results: The evaluation of the data obtained from the SF-36 revealed a reduced quality of life in the patient group in comparison to the reference population. This was particularly apparent in patients older than 60 years. The fracture healing under halo vest treatment was comparably slow. In 17% of the cases no fracture union was obtained after 12 weeks. This was only seen for patients older than 60 years. The complication rate associated to the halo vest amounted to 43% and was independent of age., Conclusion: The treatment of unstable fractures of the upper cervical spine with a halo vest results in a prolongated fracture healing for elderly people. Furthermore a halo vest therapy reduces the health related quality of life. Therefore, even for elderly patients an internal osteosynthetic stabilization of an unstable injury of the upper cervical spine should be considered if indicated.

Adrenomedullin is a multi-functional polypeptide hormone. Its involvement in angiogenesis and vasodilator action support the hypothesis that adrenomedullin may be a secretory product of neuroendocrine tumors and contribute to tumor progression. Plasma levels of adrenomedullin were measured by radioimmunoassay in 46 patients with neuroendocrine carcinomas of the gastroenteropancreatic and bronchial system. Tissue expression of adrenomedullin was studied using monoclonal antibodies on pretreated paraffin embedded tissues in a group of 31 patients. Adrenomedullin plasma levels were significantly elevated in patients compared to healthy age-matched controls (p < 0.001). The highest plasma levels were found in patients with neuroendocrine carcinomas of bronchial, midgut and unknown origin. Patients with progressive disease had higher plasma levels than patients with stable disease (p < 0.001). Of the examined tumor samples, 55 % showed cytoplasmic staining for adrenomedullin > 5 % of the total tumor area. Plasma levels and tissue expression of adrenomedullin did not correlate with functional activity of the tumors or presence of the carcinoid syndrome, but did with tumor progression (p < 0.001 and p < 0.014). In conclusion, plasma and tissue expression of the angiogenic peptide adrenomedullin are predictive of tumor progression in patients with neuroendocrine carcinomas. Adrenomedullin might represent a useful prognostic marker in patients with neuroendocrine carcinomas.

The peptide transporter PEPT2 mediates transmembrane uptake of small peptides. So far, its expression has not been evidenced in the gastrointestinal tract. We have investigated peptide transport activity in the neuromuscular layers of the gastrointestinal tract by using the fluorescent tracer-dipeptide beta-Ala-Lys-Nepsilon-7-amino-4-methyl-coumarin-3-acetic acid (Ala-Lys-AMCA). Whole-mount preparations from mouse, rat, and guinea pig stomach and small and large intestine were incubated with Ala-Lys-AMCA in the presence or absence of the uptake-inhibitors L-histidine, D-phenylalanyl-L-alanine (D-Phe-Ala), glycyl-L-sarcosine (Gly-Sar), glycyl-L-glutamine (Gly-Gln), benzylpenicillin, and cefadroxil. Fluorescence microscopy revealed that Ala-Lys-AMCA specifically accumulated in both ganglionic layers of the enteric nervous system (ENS) in all regions and species studied. This could be inhibited by Gly-Sar, D-Phe-Ala, Gly-Gln, and cefadroxil, but not by free histidine and benzylpenicillin, indicating uptake via PEPT2. Accordingly, dipeptide uptake was completely abolished in PEPT2-deficient mice. Reverse transcriptase-polymerase chain reaction analysis detected a PEPT2-specific transcript in extracts from the ganglionic ENS layers of mouse small and large intestine, further proving that enteric dipeptide transport activity is specifically mediated via PEPT2. The cellular site of dipeptide uptake was immunohistochemically localized to enteric glial cells and tissue-resident macrophages. In addition, dipeptide uptake occurred in a neurochemically defined subset of neurons in the guinea pig ENS. Our results constitute the first functional evidence for dipeptide transport activity in the ENS. PEPT2-mediated dipeptide transport in enteric glia could contribute to the clearance of neuropeptides in the ENS. In addition, the fluorophore-coupled dipeptide uptake via PEPT2 is a novel vital marker for glial cells in the ENS., (Copyright (c) 2005 Wiley-Liss, Inc.)

This study presents data on the cultural adaptation to Latino populations of two outcome measures that respond to the need for developing comprehensive instruments for outcome assessments in minority populations. We examined the psychometric properties of outcome measures designed to assess impairment in functioning, and quality of life. Impairment in functioning was measured with the Disability Assessment Schedule II (WHO-DASII) developed by the World Health Organization (1997) and quality of life was measured with A. F. Lehman's (A. F. Lehman, 1983; A. F. Lehman, 1988) shortened Quality of Life Interview (QOLI). Spanish speaking consumers (N = 198) from Fresno (CA), San Antonio (TX) and San Juan (PR) participated in this study. They were recruited from both mental health outpatient clinics and primary care rural clinics. The WHO-DASII showed good to excellent internal consistency in all sites (alpha = .72 to .97) except for one subscale (Self-Care alpha = .47). Test-retest reliability estimates were mostly moderate to substantial (.57 to .83), again with one exception, the Self-Care subscale (.46). For the QOLI internal consistency ranged from .34 to .98 and test-retest reliability ranged from .40 to .86 across all sites. An initial validation strategy using both known-groups and concurrent validity produced promising evidence of the construct validity of both measures. The Spanish versions of the WHO-DASII and the QOLI lend support to the translation and adaptation process to which these instruments were subjected.

Shear force detection is a common method of tip-sample distance control in scanning near-field optical microscopy. Shear force is the force acting on a laterally oscillating probe tip near a surface. Despite its frequent use, the nature of the interaction between tip and sample surface is a matter of debate. In order to investigate the problem, approach curves, i.e. amplitude and phase of the tip oscillation as a function of the tip-sample distance, are studied in terms of a harmonic oscillator model. The extracted force and damping constants are influenced by the substrate material. The character of the interaction ranges from elastic to dissipative. The interaction range is of atomic dimensions with a sharp onset. Between a metal-coated tip and a Cu sample, a power law for the force-distance curve is observed.

Behavioural and electrophysiological responses of Anopheles gambiae Giles sensu stricto (Diptera: Culicidae) to human skin emanations collected on glass beads were studied using a dual-port olfactometer and electroantannography. Glass beads to which skin emanations from human hands had been transferred elicited a level of attraction similar to a human hand. The attractiveness of these handled glass beads faded away 4 h after transfer onto the beads. Storage at -20 degrees C for up to 8 weeks showed a decreased but still attractive effect of the beads. In a choice test between one individual and four others, the emanations from the reference individual were significantly more attractive in three out of four cases. The headspace of handled glass beads elicited a dose-dependent EAG response. The substances causing EAG activity could be removed partially by dichloromethane, ethanol and pentane-ether. Glass beads provide a suitable neutral substrate for the transfer of human odour to enable chemical analysis of the human skin emanations for identification of kairomones of anthropophilic mosquitoes.

We studied the effects of food supplementation with Saccharomyces boulardii (S. boulardii; synonym S. cerevisiae HANSEN CBS 5926; 1 g per day for 9 days) on the presence and co-localization patterns of neuronal markers in myenteric neurones of the pig jejunum. The pan neuronal marker Hu revealed no change in the number of neuronal cell bodies per ganglion (37 +/- 7 in control vs 34 +/- 9 in the S. boulardii group). Ranked by size the following cell populations were identified: choline acetyltransferase (ChAT), calbindin-28k (CALB), substance P (SP), neurofilament 160 kD (NF-160), vasoactive intestinal polypeptide (VIP), nitric oxide synthase (NOS), calcitonin gene-related peptide (CGRP), calretinin (CALRET). We found a significant decrease in the number of CALB myenteric neurones in animals which received S. boulardii supplemented diet. None of the other neuronal markers revealed any difference between controls and S. boulardii treated animals. The study reports transmitter-localization patterns in the myenteric plexus of the pig jejunum and provides evidence that changes in the neurochemistry of enteric neurones occur with S. boulardii supplemented diet. Although only CALB expression was altered and the functional significance of this finding remains unknown, our study identified a possible new effector level of probiotics in the gut.

During recent years, quality of life has become an increasingly important parameter in the outcome assessment of therapeutic protocols in oncology. In rectal cancer treatment, the influence of different surgical techniques on quality of life can be evaluated and compared by a variety of standardized and clinically validated instruments. As yet, sphincter-saving surgical techniques were considered to result in higher quality of life than those sacrificing the sphincter. Recent data from clinical trials indicates that stoma patients have comparable or even higher quality of life than patients of sphincter-saving surgery. However, prospective clinical trials assessing not only classic quality of life items but also unconventional parameters, i.e., patient expectations etc., are still lacking. Thus, detailed interpretation of the differences assessed by the classic quality of life scores is profoundly hampered. The realisation of such a prospective clinical trial will be a further important step to improve quality of life in patients suffering from rectal cancer by improving not only surgical techniques but also appropriate psychological patient management.

In this paper we report on the process of translating five mental health outcome measures into Spanish and adapting them to Latino culture. The instruments considered are the World Health Organization-Disability Assessment Scale, the Burden Assessment Scale, the Family Burden Scale, Lehman's Quality of Life Interview and the Continuity of Care in Mental Health Services Interview. A systematic process of translation and adaptation of the instruments was followed with the goal of achieving cultural equivalence between the English and Spanish versions of the instruments in five dimensions: semantic, content, technical, construct, and criterion equivalence. In this paper we present data about the semantic, content, and technical equivalence. Various steps were taken to achieve equivalence in these dimensions, including the use of a bilingual committee, a multi-national bilingual committee, back-translation, and focus groups with mental health patients and their relatives.

The intrinsic innervation of muscle layers in the mammalian gastrointestinal tract has been mainly studied in nonruminants. The aim of this study was to identify intrinsic motor neurones in the bovine abomasum that innervate the circular and longitudinal muscles. Circular (CMN) and longitudinal muscle motor neurones (LMN) were selectively labeled by application of the retrograde tracer 1,1'-didodecyl-3,3,3',3'-tetramethyl indocarbocyanine perchlorate (DiI) onto the muscle layers. The transmitter phenotype was determined by immunohistochemical detection of choline acetyltransferase (ChAT), nitric oxide synthase (NOS), and neurone-specific enolase (NSE). On average, the myenteric ganglia contained 61 +/- 19 NSE-positive cell bodies, of which 89% were ChAT-positive and 10% were NOS-positive. Only 0.7% of NSE-positive neurones (41 of 5,777) contained both ChAT and NOS. Application of DiI onto the circular and longitudinal muscles revealed on average 60 +/- 27 (n = 4) and 68 +/- 36 (n = 4), respectively, labeled cell bodies in the myenteric plexus. For the circular and longitudinal muscles the proportions of ascending to descending neurones were 76 : 24% and 54 : 46%, respectively. While most ascending CMN were ChAT-positive (96%), 51% of the descending CMN were ChAT-negative. All ascending and 95% of descending LMN were ChAT-positive. It was concluded that cholinergic excitatory innervation is predominant in both muscle layers of the abomasum. Whereas the circular muscle receives cholinergic excitatory and nitrergic inhibitory innervation, the longitudinal muscle is only innervated by cholinergic pathways. This innervation pattern is different from that in gastric muscle layers in monogastric animals., (Copyright 2002 Wiley-Liss, Inc.)

Knowledge of the neurochemical coding of submucosal neurones in the human gut is important to assess neuronal changes under pathological conditions. We therefore investigated transmitter colocalization patterns in rectal submucosal neurones in normal tissue (n=11) and in noninflamed tissue of Crohn's disease (CD) patients (n=17). Neurone-specific enolase (NSE), choline acetyltransferase (ChAT), vasoactive intestinal polypeptide (VIP), substance P (SP), nitric oxide synthase (NOS) and calcitonin gene-related peptide (CGRP) were detected immunohistochemically in whole-mount preparations from rectal biopsies. The neuronal marker NSE revealed no differences in the number of cells per ganglion (controls 5.0; CD 5.1). Four cell populations with distinct neurochemical codes were identified. The sizes of the populations ChAT/VIP (58% vs. 55%), ChAT/SP (8% vs. 8%), and ChAT/- (22% vs. 22%) were similar in control and CD. The population VIP/- was significantly increased in CD (12% vs. 2% in controls). Unlike in controls, all NOS neurones colocalized ChAT in CD. Thickened CGRP-fibres occurred in CD. We identified neurochemically distinct populations in the human submucous plexus. The increase in the VIP/- population, extensive colocalization of ChAT and NOS and hypertrophied CGRP fibres indicated adaptive changes in the enteric nervous system in noninflamed rectum of CD patients.

We recently described calbindin immunoreactivity in the myenteric plexus of the guinea-pig stomach. To study the neurochemical coding of calbindin D28 k (CALB)-containing myenteric neurones, the presence of calretinin (CALRET), choline acetyltransferase (ChAT), enkephalin (ENK), neuropeptide Y, serotonin (5-HT), somatostatin (SOM) and substance P(SP) was investigated immunohistochemically in colchicine-treated preparations. Nitric oxide synthase-containing neurones were detected by NADPH-diaphorase histochemistry. In addition, we investigated the neurone distribution patterns around the gastric corpus. Most CALB neurones were ChAT positive. ChAT/CALB neurones were either CALRET (ca 75%) or 5-HT positive and most contained in addition SP and/or ENK. All 5-HT neurones contained CALB. CALB labelled on average 2.3, 4.8 and 7.5 neurones per ganglion at the lesser curvature, in the central region and the greater curvature, respectively, which indicated a preferential localisation at the greater curvature. Compared to the total number of myenteric neurones, the proportion of CALB neurones increased significantly from the lesser curvature (6%) towards the greater curvature (18%). This shift, although observed for most ChAT/CALB-positive populations, was most prominent for the ChAT/CALB/CALRET/SP/ENK-encoded neurones. SOM-positive and ChAT-only encoded neurones were preferentially located at the lesser curvature. The remaining ten neurochemically defined populations did not exhibit an uneven distribution. The colocalisation of CALB with CALRET or 5-HT is specific for myenteric neurones in the stomach and represents one significant difference to the neurochemical code of CALB neurones in the guinea-pig intestine. The functional significance of the unevenness of neurone distribution along the circumference of the gastric corpus remains to be studied.

Exposing [Bi(OR)3(toluene)]2 (1, R = OC6F5) to different solvents leads to the formation of larger polymetallic bismuth oxo alkoxides via ether elimination/oligomerization reactions. Three different compounds were obtained depending upon the conditions: Bi4(mu 4-O)(mu-OR)6(mu 3-OBi(mu-OR)3)2(C6H5CH3) (2), Bi8(mu 4-O)2(mu 3-O)2(mu 2-OR)16 (3), Bi6(mu 3-O)4(mu 3-OR)(mu 3-OBi(OR)4)3 (4). Compounds 2 and 3 can also be synthesized via an alcoholysis reaction between BiPh3 and ROH in refluxing dichloromethane or chloroform. Related oxo complexes NaBi4(mu 3-O)2(OR)9(THF)2 (5) and Na2Bi4(mu 3-O)2(OR)10(THF)2 (6) were obtained from BiCl3 and NaOR in THF. The synthesis of 1 and Bi(OC6Cl5)3 via salt elimination was successful when performed in toluene as solvent. For compounds 2-6 the single-crystal X-ray structures were determined. Variable-temperature NMR spectra are reported for 2, 3, and 5.

The goal of this report is to summarise the current knowledge on the projection pathways of enteric neurones innervating the muscle and mucosa in different regions of the gut. Combination of neuronal tracing, immunohistochemical and electrophysiological methods has allowed researchers to gain insight into the enteric hardwiring of specific target tissue in the gut. A polarised innervation pattern of the circular muscle was demonstrated for the stomach fundus/corpus and the ileum with descending pathways being primarily nitrergic while ascending pathways were primarily cholinergic. This characteristic hardwiring is thought to set in part the functional basis for peristalsis. A similar polarised innervation pathway was found for the enteric innervation of the mucosa in the stomach and large intestine but not in the small intestine. In both the stomach (myenteric neurones) and in the proximal and distal colon (submucosal neurones), ascending pathways to the mucosa are primarily cholinergic while descending pathways are primarily non-cholinergic. In the colon, results suggest that activation of both pathways induces a cross potentiation of cholinergic and vasoactive intestinal polypeptidergic mediated secretion. Furthermore, a large population of myenteric neurone s projecting to the mucosa in the small and large intestine are probably intrinsic primary afferent neurones sensitive to mechanical as well as chemical stimuli.

Using immunohistochemistry we studied the presence of calbindin in myenteric neurones of the guinea-pig stomach. A rabbit anti recombinant rat calbindin-D28k (CALB) stained 12, 12 and 25% of all myenteric neurones in the fundus, corpus and antrum, respectively. A rabbit anti recombinant human CALB stained 4, 4 and 16%, respectively. A mouse monoclonal antibody against the chicken intestinal CALB showed no labelling. In all regions most calbindin neurones were additionally choline acetyltransferase (ChAT) positive while only a small proportion exhibited nicotinamide adenosine dinucleatide phosphate (NADPH)-diaphorase-activity. Numerous calbindin-positive varicose nerve fibres were present within myenteric ganglia, rarely detectable in the muscle layers and virtually absent in the mucosa. This study demonstrated that a supopulation of cholinergic myenteric neurones in the stomach contain calbindin and suggested that many of these neurones fulfil interneuronal tasks.