Dose-response effect of interleukin (IL)-1beta, tumour necrosis factor (TNF)-alpha, and interferon-y on the in vitro production of epithelial neutrophil activating peptide-78 (ENA-78), IL-8, and IL-6 by human endometrial stromal cells

The production of epithelial neutrophil activating peptide-78 (NA-78) and the interleukins IL-8 and IL-6 by endometrial stromal cells is stimulated by pro-inflammatory interleukin-1 (IL-1) and tumour necrosis factor-α (TNF-α). IL-8 is suggested to play a role in the pathogenesis of endometriosis, and in these women the peritoneal fluid concentrations of ENA-78 and IL-8 are increased. TNF-α has been tested together with interferon-γ because of their cooperative stimulation of IL-6. The release of IL-8, however, is inhibited with increasing interferon levels. The aim of the study was the analysis of the production of ENA-78, IL-6 and IL-8 by cultured human endometrial stromal cells in the presence of varying concentrations of IL-1β, TNF-α, and interferon-γ.

Response, survival, and long-term toxicity after therapy with the radiolabeled somatostatin analogue [90Y-DOTA]-TOC in metastasized neuroendocrine cancers

To investigate response, survival, and safety profile of the somatostatin-based radiopeptide (90)yttrium-labeled tetraazacyclododecane-tetraacetic acid modified Tyr-octreotide ([(90)Y-DOTA]-TOC) in neuroendocrine cancers.

Germania: l’indomabile trading state

Basato su interviste con i principali attori tedeschi e su un’analisi della letteratura, l’articolo analizza lo sviluppo recente dell’economia tedesca e la strategia tedesca nell’affrontare la crisi dell’eurozona. La Germania è uno stato commerciale (trading state), la cui crescita è fortemente trainata dalle esportazioni. Fino agli anni novanta, rigidità istituzionali forti, nel sistema di relazioni industriali e nel sistema di protezione sociale, contribuivano a conciliare lo sviluppo delle esportazioni con una crescita armonica dei consumi interni, contribuendo cosi a ingabbiare la «tigre» tedesca. A partire dagli anni novanta, sia le relazioni industriali sia la protezione sociale sono state fortemente liberalizzate, stimolando ulteriormente la competitività estera e indebolendo i consumi interni. Il modello economico tedesco, cosi come è venuto profilandosi negli ultimi dieci anni, è alla base delle politiche di austerità che la Germania impone all’Europa. Tali politiche sono fortemente condivise dai partiti politici, dagli attori sociali e dall’opinione pubblica, e le probabilità che la strategia tedesca cambi sono minime.

Climate warming and vegetation response after Heinrich event 1 (16 700–16 000 cal yr BP) in Europe south of the Alps

Chironomids preserved in a sediment core from Lago di Origlio (416 m a.s.l.), a lake in the foreland of the Southern Swiss Alps, allowed quantitative reconstruction of Late Glacial and Early Holocene summer temperatures using a combined Swiss–Norwegian temperature inference model based on chironomid assemblages from 274 lakes. We reconstruct July air temperatures of ca. 10 °C between 17 300 and 16 000 cal yr BP, a rather abrupt warming to ca. 12.0 °C at ca. 16 500–16 000 cal yr BP, and a strong temperature increase at the transition to the Bølling/Allerød interstadial with average temperatures of about 14 °C. During the Younger Dryas and earliest Holocene similar temperatures are reconstructed as for the interstadial. The rather abrupt warming at 16 500–16 000 cal yr BP is consistent with sea-surface temperature as well as speleothem records, which indicate a warming after the end of Heinrich event 1 (sensu stricto) and before the Bølling/Allerød interstadial in southern Europe and the Mediterranean Sea. Pollen records from Origlio and other sites in southern Switzerland and northern Italy indicate an early reforestation of the lowlands 2000–1500 yr prior to the large-scale afforestation of Central Europe at the onset of the Bølling/Allerød period at ca. 14 700–14 600 cal yr BP. Our results suggest that these early afforestation processes in the formerly glaciated areas of northern Italy and southern Switzerland have been promoted by increasing temperatures.

Sensitivity to endothelin-1 is decreased in isolated livers of endothelial constitutive nitric oxide synthase knockout mice

ABSTRACT: BACKGROUND: Hepatic sinusoidal resistance is regulated by vasoactive factors including endothelin-1 (ET-1) and nitric oxide (NO). In the absence of NO, vasoconstrictor response to endothelin is expected to predominate. Therefore, we hypothesized sensitivity to endothelin to be increased in mice lacking the endothelial cell NO synthase gene. Response of vascular resistance to endothelin was assessed in the in situ perfused liver of endothelial constitutive nitric oxide synthase (ecNOS) knockout and wild type mice. Livers were also harvested for RNA and protein isolation for quantitative PCR and Western blotting, respectively. The expression of endothelin receptors, isoenzymes of NO synthase, heme-oxygenase and adrenomedullin was quantified. RESULTS: Endothelin increased hepatic vascular resistance in a dose-dependent manner in both strains; however, this increase was significantly less in ecNOS knockout mice at physiologic concentrations. Expression of heme-oxygenases and adrenomedullin was similar in both groups, whereas inducible nitric oxide synthase (iNOS) protein was not detectable in either strain. mRNA levels of pre-pro-endothelin-1 and ETB receptor were comparable in both strains, while mRNA for ETA receptor was decreased in ecNOS knockouts. CONCLUSION: Livers of ecNOS knockout mice have a decreased sensitivity to endothelin at physiologic concentrations; this is associated with a decreased expression of ETA receptors, but not with other factors, such as iNOS, ETB receptors, adrenomedullin or heme-oxygenase. Further studies targeting adaptive changes in ETA receptor distribution and/or intracellular signaling downstream of the receptor are indicated.

Virus isolation vs RT-PCR: which method is more successful in detecting VHSV and IHNV in fish tissue sampled under field conditions?

This study compared the results of reverse transcription-polymerase chain reaction (RT-PCR) and traditional virus isolation on cell culture in detection of viral haemorrhagic septicaemia virus (VHSV) and infectious haematopoietic necrosis virus (IHNV). RT-PCR was used for 172 tissue sample pools (total of 859 fish) originating from a field survey on the occurrence of VHSV and IHNV in farmed and wild salmonids in Switzerland. These samples represented all sites with fish that were either identified as virus-positive by means of virus isolation (three sites, four positive tissue sample pools) and/or demonstrated positive anti-VHSV-antibody titres (83 sites, 121 positive blood samples) in a serum plaque neutralization test (SPNT). The RT-PCR technique confirmed the four VHSV-positive tissue sample pools detected by virus isolation and additionally identified one VHSV-positive sample that showed positive anti-VHSV-AB titres, but was negative in virus isolation. With IHNV, RT-PCR detected two positive samples not identified by virus isolation while in these fish the SPNT result had been questionable. One of the IHNV-positive samples represents the first detection of IHNV-RNA in wild brown trout in Switzerland. Compared to SPNT, the RT-PCR method detected, as with virus isolation, a much lower number of positive cases; reasons for this discrepancy are discussed. Our results indicate that RT-PCR can not only be successfully applied in field surveys, but may also be slightly more sensitive than virus isolation. However, in a titration experiment under laboratory conditions, the sensitivity of RT-PCR was not significantly higher when compared with virus isolation.

Expression profiling with RNA from formalin-fixed, paraffin-embeeded material

Background Molecular characterization of breast and other cancers by gene expression profiling has corroborated existing classifications and revealed novel subtypes. Most profiling studies are based on fresh frozen (FF) tumor material which is available only for a limited number of samples while thousands of tumor samples exist as formalin-fixed, paraffin-embedded (FFPE) blocks. Unfortunately, RNA derived of FFPE material is fragmented and chemically modified impairing expression measurements by standard procedures. Robust protocols for isolation of RNA from FFPE material suitable for stable and reproducible measurement of gene expression (e.g. by quantitative reverse transcriptase PCR, QPCR) remain a major challenge. Results We present a simple procedure for RNA isolation from FFPE material of diagnostic samples. The RNA is suitable for expression measurement by QPCR when used in combination with an optimized cDNA synthesis protocol and TaqMan assays specific for short amplicons. The FFPE derived RNA was compared to intact RNA isolated from the same tumors. Preliminary scores were computed from genes related to the ER response, HER2 signaling and proliferation. Correlation coefficients between intact and partially fragmented RNA from FFPE material were 0.83 to 0.97. Conclusion We developed a simple and robust method for isolating RNA from FFPE material. The RNA can be used for gene expression profiling. Expression measurements from several genes can be combined to robust scores representing the hormonal or the proliferation status of the tumor.

Das Burnout-Syndrom: eine medizinische Perspektive

The burnout syndrome is a heterogeneous concept mostly understood as a complex of symptoms, primarily exhaustion, in response to prolonged emotional and interpersonal stress at work. The prevalence of burnout is considerably high in Swiss primary care physicians. In spite of its vague definition, burnout is a serious stress disease with many associated medical problems and high economic costs. Previous recommendations for the psychosomatic management of patients with functional somatic syndromes also apply to burnout treatment. These are complemented by more specific interventions targeting job stress related factors. Relapse prevention focuses on early recognition of warning signs and is an ongoing process.

Cardiovascular response to beta-adrenergic blockade or activation in 23 inbred mouse strains

We report the characterisation of 27 cardiovascular-related traits in 23 inbred mouse strains. Mice were phenotyped either in response to chronic administration of a single dose of the beta-adrenergic receptor blocker atenolol or under a low and a high dose of the beta-agonist isoproterenol and compared to baseline condition. The robustness of our data is supported by high trait heritabilities (typically H(2)>0.7) and significant correlations of trait values measured in baseline condition with independent multistrain datasets of the Mouse Phenome Database. We then focused on the drug-, dose-, and strain-specific responses to beta-stimulation and beta-blockade of a selection of traits including heart rate, systolic blood pressure, cardiac weight indices, ECG parameters and body weight. Because of the wealth of data accumulated, we applied integrative analyses such as comprehensive bi-clustering to investigate the structure of the response across the different phenotypes, strains and experimental conditions. Information extracted from these analyses is discussed in terms of novelty and biological implications. For example, we observe that traits related to ventricular weight in most strains respond only to the high dose of isoproterenol, while heart rate and atrial weight are already affected by the low dose. Finally, we observe little concordance between strain similarity based on the phenotypes and genotypic relatedness computed from genomic SNP profiles. This indicates that cardiovascular phenotypes are unlikely to segregate according to global phylogeny, but rather be governed by smaller, local differences in the genetic architecture of the various strains.

Canine keratinocytes upregulate type I interferons and proinflammatory cytokines in response to poly(dA:dT) but not to canine papillomavirus.

Papillomaviruses (PV) are double stranded (ds) DNA viruses that infect epithelial cells within the skin or mucosa, most often causing benign neoplasms that spontaneously regress. The immune system plays a key role in the defense against PVs. Since these viruses infect keratinocytes, we wanted to investigate the role of the keratinocyte in initiating an immune response to canine papillomavirus-2 (CPV-2) in the dog. Keratinocytes express a variety of pattern recognition receptors (PRR) to distinguish different cutaneous pathogens and initiate an immune response. We examined the mRNA expression patterns for several recently described cytosolic nucleic acid sensing PRRs in canine monolayer keratinocyte cultures using quantitative reverse transcription-polymerase chain reaction. Unstimulated normal cells were found to express mRNA for melanoma differentiation associated gene 5 (MDA5), retinoic acid-inducible gene I (RIG-I), DNA-dependent activation of interferon regulatory factors, leucine rich repeat flightless interacting protein 1, and interferon inducible gene 16 (IFI16), as well as their adaptor molecules myeloid differentiation primary response gene 88, interferon-β promoter stimulator 1, and endoplasmic reticulum-resident transmembrane protein stimulator of interferon genes. When stimulated with synthetic dsDNA [poly(dA:dT)] or dsRNA [poly(I:C)], keratinocytes responded with increased mRNA expression levels for interleukin-6, tumor necrosis factor-α, interferon-β, RIG-I, IFI16, and MDA5. There was no detectable increase in mRNA expression, however, in keratinocytes infected with CPV-2. Furthermore, CPV-2-infected keratinocytes stimulated with poly(dA:dT) and poly(I:C) showed similar mRNA expression levels for these gene products when compared with expression levels in uninfected cells. These results suggest that although canine keratinocytes contain functional PRRs that can recognize and respond to dsDNA and dsRNA ligands, they do not appear to recognize or initiate a similar response to CPV-2.

Impact of cardiac computed tomography of the interatrial septum before pulmonary vein isolation

BACKGROUND Multidetector computed tomography (MDCT) may be useful to identify patients with patent foramen ovale (PFO). The aim of this study was to analyze whether a MDCT performed before pulmonary vein isolation reliably detects a PFO that may be used for access to the left atrium. METHODS AND RESULTS In 79 consecutive patients, who were referred for catheter ablation of symptomatic paroxysmal or persistent atrial fibrillation (AF), the presence of a PFO was explored by MDCT and transesophageal echocardiography (TEE). TEE was considered as the gold standard, and quality of TEE was good in all patients. In 16 patients (20.3%), MDCT could not be used for analysis because of artifacts, mainly because of AF. On TEE, a PFO was found in 15 (23.8%) of the 63 patients with usable MDCT. MDCT detected six PFO of which four were present on TEE. This corresponded to a sensitivity of 26.7%, a specificity of 95.8%, a negative predictive value of 80.7%, and a positive predictive value of 66.7%. The receiver operating characteristics curve of MDCT for the detection of PFO was 0.613 (95% confidence interval 0.493-0.732). CONCLUSIONS MDCT may detect a PFO before pulmonary isolation. However, presence of AF may lead to artifacts on MDCT impeding a meaningful analysis. Furthermore, in this study sensitivity and positive predictive value of MDCT were low and therefore MDCT was not a reliable screening tool for detection of PFO.

Asking Sensitive Questions in Online Surveys: An Experimental Comparison of the Randomized Response Technique and the Crosswise Model

Self-administered online surveys provide a higher level of privacy protection to respondents than surveys administered by an interviewer. Yet, studies show that asking sensitive questions is problematic also in self-administered mode. Because respondents might not be willing to reveal the truth and provide answers that are subject to social desirability bias, the validity of prevalence estimates of sensitive behaviors gained via online surveys can be challenged. A well-known method to combat these problems is the Randomized Response Technique (RRT). However, convincing evidence that the RRT provides more valid estimates than direct questioning in online mode is still lacking. Moreover, an alternative approach called the Crosswise Model (CM) has recently been suggested to overcome some of the deficiencies of the RRT. In the context of an online survey on plagiarism and cheating on exams among students of two Swiss universities (N = 6,494), we tested different implementations of the RRT and the CM and compared them to direct questioning using a randomized experimental design. Results reveal a poor performance of the RRT, which failed to elicit higher prevalence estimates than direct questioning. Using the CM however, significantly higher prevalence estimates were obtained making it a promising new alternative to the conventional RRT.

Metabolic adaptations and changes of the mammary immune response during beta-hydroxybutyrate administration

Elevation of ketone bodies occurs frequently after parturition during negative energy balance in high yielding dairy cows. Previous studies illustrated that hyperketonemia interferes with metabolism and it is assumed that it impairs the immune response. However, a causative effect of ketone bodies could not be shown in vivo before, because spontaneous hyperketonemia comes usually along with high NEFA and low glucose concentrations. The objective was to study effects of beta-hydroxybutyrate (BHBA) infusion and an additional intramammary lipopolysaccharide (LPS) challenge on metabolism and immune response in dairy cows. Thirteen dairy cows received intravenously either a BHBA infusion (group BHBA, n=5) to induce hyperketonemia (1.7 mmol/L), or an infusion with a 0.9 % saline solution (Control, n=8) for 56 h. Infusions started at 0900 on day 1 and continue up to 1700 two days later. Two udder quarters were challenged with 200 μg Escherichia coli-LPS 48 h after the start of infusion. Blood samples were taken one week and 2 h before the start of infusions as reference samples and hourly during the infusion. Liver and mammary gland biopsies were taken one week before the start of the infusion, 48 h after the start of the infusion, and mammary tissues was additionally taken 8 h after LPS challenge (56 h after the start of infusions). Rectal temperature (RT) and somatic cell count (SCC) was measured before and 48 h after the start of infusions and hourly during LPS challenge. Blood samples were analyzed for plasma glucose, BHBA, NEFA, triglyceride, urea, insulin, glucagon, and cortisol concentration. The mRNA abundance of factors related to potential adaptations of metabolism and immune system was measured in liver and mammary tissue biopsies. Differences between blood constituents, RT, SCC, and mRNA abundance before and 48 h after the start of infusions, and differences between mRNA abundance before and after LPS challenges were tested for significance by GLM of SAS procedure with treatment as fixed effect. Area under the curve was calculated for blood variables during 48 h BHBA infusion and during the LPS challenge, and additionally for RT and SCC during the LPS challenge. Most surprisingly, both plasma glucose and glucagon concentration decreased during the 48 h of BHBA infusion (P<0.05). During the 48 h of BHBA infusion, serum amyloid A mRNA abundance in mammary gland was increased (P<0.01), and haptoglobin (Hp) mRNA abundance tended to increase in cows treated with BHBA compared to control group (P= 0.07). RT, SCC, and candidate genes related to immune response in the liver were not affected by BHBA infusion. However, during LPS challenge the expected increase of both plasma glucose and glucagon concentration was much less pronounced in the animals treated with BHBA (P<0.05) and also SCC increased much less pronounced in the animals infused with BHBA (P<0.05) than in the controls. An increased BHBA infusion rate to maintain plasma BHBA constant could not fully compensate for the decreased plasma BHBA during the LPS challenge which indicates that BHBA is used as an energy source during the immune response. In addition, BHBA infused animals showed a more pronounced increase of mRNA abundance of IL-8, IL-10, and citrate synthase in the mammary tissue of LPS challenged quarters (P<0.05) than control animals. Results demonstrate that infusion of BHBA affects metabolism through decreased plasma glucose concentration which is likely related to a decreased release of glucagon during hyperketonemia and during additional inflammation. It also affects the systemic and mammary immune response which may reflect the increased susceptibility for mastitis during spontaneous hyperketonemia. The obviously reduced gluconeogenesis in response to BHBA infusion may be a mechanism to stimulated the use of BHBA as an energy source instead of glucose, and/or to save oxaloacetate for the citric acid cycle instead of gluconeogenesis and as a consequence to reduce ketogenesis.