The B-type natriuretic peptide of the Congo and Timneh grey parrot.

In captivity, cardiovascular diseases are common in grey parrots. The diagnosis of these diseases in living birds is difficult, and new diagnostic possibilities would be desirable. The heart is an important endocrine organ in which cardiomyocytes synthetise B-type natriuretic peptide (BNP) and release it into the bloodstream. This hormone has a significant role in cardiovascular and body fluid regulation. The blood concentration of BNP is used in human medicine and small animal medicine as a diagnostic tool in the identification of heart diseases and as a prognostic marker for the risk of mortality. The nucleotide and amino acid sequence of BNP was described in Congo (n = 4) and Timneh (n = 3) grey parrots by PCR after RNA isolation from the atria and ventricles. The results showed a high similarity between the nucleotide sequences of the grey parrots' BNP and the already known sequence of this hormone in chickens. The amino acid sequence of the mature peptide region is consistent in these three species. BNP plasma concentration could be a possible blood parameter for identifying clinically manifest cardiovascular diseases in grey parrots as it is in other species.

Alpha-fetoprotein promoter-targeted sodium iodide symporter gene therapy of hepatocellular carcinoma.

Due to limited treatment options the prognosis of patients with advanced hepatocellular cancer (HCC) has remained poor. To investigate an alternative therapeutic approach, we examined the feasibility of radioiodine therapy of HCC following human sodium iodide symporter (NIS) gene transfer using a mouse alpha-fetoprotein (AFP) promoter construct to target NIS expression to HCC cells. For this purpose, the murine Hepa 1-6 and the human HepG2 hepatoma cell lines were stably transfected with NIS cDNA under the control of the tumor-specific AFP promoter. The stably transfected Hepa 1-6 cell line showed a 10-fold increase in iodide accumulation, while HepG2 cells accumulated (125)I approximately 60-fold. Tumor-specific NIS expression was confirmed on mRNA level by northern blot analysis, and on protein level by immunostaining, that revealed primarily membrane-associated NIS-specific immunoreactivity. In an in vitro clonogenic assay up to 78% of NIS-transfected Hepa 1-6 and 93% of HepG2 cells were killed by (131)I exposure, while up to 96% of control cells survived. In vivo NIS-transfected HepG2 xenografts accumulated 15% of the total (123)I administered per gram tumor with a biological half-life of 8.38 h, resulting in a tumor absorbed dose of 171 mGy MBq(-1) (131)I. After administration of a therapeutic (131)I dose (55.5 MBq) tumor growth of NIS expressing HepG2 xenografts was significantly inhibited. In conclusion, tumor-specific iodide accumulation was induced in HCC cells by AFP promoter-directed NIS expression in vitro and in vivo, which was sufficiently high to allow a therapeutic effect of (131)I. This study demonstrates the potential of tumor-specific NIS gene therapy as an innovative treatment strategy for HCC.

An error model for protein quantification.

MOTIVATION: Quantitative experimental data is the critical bottleneck in the modeling of dynamic cellular processes in systems biology. Here, we present statistical approaches improving reproducibility of protein quantification by immunoprecipitation and immunoblotting. RESULTS: Based on a large data set with more than 3600 data points, we unravel that the main sources of biological variability and experimental noise are multiplicative and log-normally distributed. Therefore, we suggest a log-transformation of the data to obtain additive normally distributed noise. After this transformation, common statistical procedures can be applied to analyze the data. An error model is introduced to account for technical as well as biological variability. Elimination of these systematic errors decrease variability of measurements and allow for a more precise estimation of underlying dynamics of protein concentrations in cellular signaling. The proposed error model is relevant for simulation studies, parameter estimation and model selection, basic tools of systems biology. AVAILABILITY: Matlab and R code is available from the authors on request. The data can be downloaded from our website www.fdm.uni-freiburg.de/~ckreutz/data.

[The hepatocellular carcinoma: alternative therapeutical strategies]. / Das hepatozelluläre Karzinom: alternative Therapiestrategien.

Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in some areas of the world. The prognosis of HCC patients is generally very poor with a 5-year survival rate of less than 5%. Therapeutic strategies include surgery (resection or liver transplantation) and non-surgical interventions, such as percutaneous ethanol injection or radiofrequency thermal ablation as well as transarterial embolization or chemoembolization. Therefore, the development and evaluation of novel HCC treatment strategies such as the use of antiangiogenic, antiproliferative or antiinflammatoric drugs, immune therapeuticals, gene therapy and internal or external radiation are of utmost importance. This review should give an overview of possible alternative therapies in HCC treatment.

[Hepatocellular carcinoma: novel therapeutic approaches]. / Hepatozelluläres Karzinom: Innovative Therapieansätze.

Therapeutic options for patients with hepatocellular carcinoma (HCC) are still limited. Only patients with small HCC are considered for curative treatment options such as resection, liver transplantation or local ablation. Systemic treatment of advanced HCC is still impaired by the high level of resistance to chemotherapy and irradiation. In addition, most patients with HCC are highly susceptible to toxic side effects due to their impaired liver function. Novel treatment options are based on the increasing understanding of molecular mechanisms involved in tumorigenesis, tumor progression and metastasis. Such "targeted therapies" aim at growth factors and their receptors, intracellular signal transduction and cell cycle control. Anti-angiogenesis targets the tumor vasculature. Gene therapeutic strategies are based on the transfer of therapeutic genes in target tissues to express highly active proteins such as suicide genes or cytokines. Tumor cell death by cell lysis is the mechanism of oncolytic viruses, which are able to selectively replicate in tumor cells. This mechanism may be augmented by the addition of therapeutic genes expressed by recombinant oncolytic viruses. The induction of a potent immune response to tumor antigens by vaccination strategies or by adoptive transfer of activated immune cells aim at the specific destruction of tumor cells by effector T-cells targeted to tumor antigens.

[Regional therapy of liver tumors]. / Regionale Therapie von Lebertumoren.

Malignant liver tumors are either originating from the liver, such as the primary liver tumors hepatocellular carcinoma and the cholangiocellular carcinoma, or metastases from extrahepatic malignancies. Apart from surgical procedures (resection, liver transplantation) percutaneous local-ablative (ethanol injection, radiofrequency thermal ablation as well as radiation therapy) and transarterial interventions are non-surgical therapeutic options. While these regional therapies have been shown in randomised controlled studies to be effective for hepatocellular carcinoma, their therapeutic efficacy in cholangiocellular carcinoma and liver metastases has not been shown. In the following we will summarize the regional therapeutic options in primary and secondary liver tumors.

[Seizures following Billroth II gastrectomy]. / Krampfanfall nach Billroth-II-Operation.

HISTORY: A 64-year old somnolent man was admitted to the emergency department with a reported seizure half an hour earlier. Due to similar episodes the patient had been treated with antiepileptics in the past. The patient s past history revealed a partial gastrectomy (Billroth II) more than ten years ago. DIAGNOSTIC FINDINGS AND THERAPY: At the time of admission blood glucose was 31 mg/dl. Other routine laboratory analyses and the clinical examination were normal. In addition, a detailed neurological examination and a cranial CT-scan were normal. Due to the hypoglycemia a dumping syndrome was suspected. A three hour oral glucose tolerance test (OGTT) resulted in a late hypoglycemia, establishing the diagnosis of late dumping. After adaptation of the patient's diet no further hypoglycemic episodes occurred. CONCLUSION: Manifestation of a dumping syndrome may occur even years after gastrectomy. Therefore, in patients presenting with hypoglycemia and a history of gut surgery, a dumping syndrome should be suspected. Furthermore, seizures due to hypoglycemia may be the only manifestation of late dumping.

[Local ablation of malignant lesions of the liver - potential applications and limitations of the different methods]. / Lokale Ablationsverfahren als Ergänzung der Therapie maligner Leberläsionen.

Malignant lesions of the liver are among the most frequent and difficult problems in clinical practice. Liver tumors can be classified as primary (hepatocellular carcinoma HCC, cholangiocarcinoma CC) and secondary liver lesions (metastasis). The therapeutic principle of resecting liver tumours, applies to both types of liver lesions. Unfortunately many patients with primary and secondary liver lesions are inoperable because of technical difficulties or comorbidity. This stimulated the development of percutaneous ablation methods. These procedures allow to destroy tumors percutaneously with alcohol, acetic acid, radiofrequency, microwaves or laser. Potential applications and limitations of the methods are discussed.

Peer support groups in multiple sclerosis: current effectiveness and future directions.

Peer support programs have become a common method of providing support for patients with chronic illness. Utilizing peers as resources has been proposed as an effective means for coping with a stressful life experience and for gaining support from others who share a common factor, although data are somewhat mixed on the efficacy of peer support. The aim of the present study was to evaluate the effectiveness of eight weeks of a standard form of peer support in improving quality of life and reducing depressive symptoms in 44 patients with multiple sclerosis (MS). One person from each of six groups participated in a training course in order to learn basic principles of peer support. Eight weekly sessions were held and patients completed self-administered questionnaires pre- and post-treatment assessing quality of life and depression. Results showed that support groups do not provide consistent improvement in quality of life or depression in patients with MS and suggest that patients who have better mental health functioning could be at risk for deterioration in support groups.

Immunobiology and gene-based immunotherapy of hepatocellular carcinoma.

PURPOSE: Hepatocellular carcinoma (HCC) is one of the major malignancies worldwide. For most patients with advanced or multifocal HCC treatment options are limited resulting in a poor prognosis. Several local ablation methods have been developed as minimally invasive strategies for HCC treatment. It is unclear, until now, whether these therapies will significantly improve the poor prognosis of patients with unresectable HCC. Novel therapeutic strategies and a better understanding of HCC imunobiology are, therefore, urgently required. DESIGN: The scientific literature since 1970 in all languages cited in Medline was systematically reviewed. RESULTS: Until now, a variety of specific and non-specific immunostimulatory strategies against HCC has been applied in preclinical experimental models with some promising results. The molecular characterization of HCC associated tumour antigens such as alpha-fetoprotein (AFP) and the increased understanding of the immunological pathways involved in liver and tumor immunology have paved the way for the design of promising gene-based cancer vaccines. The first phase I and II immunotherapeutic clinical trials based on dendritic cell immunotherapy and peptide vaccines are ongoing in HCC-patients. Clinical trials have, in general, demonstrated the safety of such strategies. Recently, exciting new immunological techniques and tools have been developed which allow to characterize antigen specific T cells at a single-cell level. In future, HCC specific tumor rejection antigens which can be used therapeutically have to be identified using microarray-based analysis. The different therapeutic modalities need to be compared directly resulting in optimised therapeutic approaches and the identification of sub-groups of HCC-patients responding favourably to treatment.

Stable chromosome aberrations and ionizing radiation in airline pilots.

INTRODUCTION: The purpose of this study was to determine the frequency of translocations and insertions in the blood of long-term pilots in relation to estimated cumulative radiation dose received while flying, and to compare that to the frequency in a group of similarly aged men without a history of frequent airline travel. METHODS: Healthy, non-smoking male pilots aged 40-60 yr were recruited from a single airline. Non-pilot controls were recruited from healthy, non-smoking professional males in the same age range and without a history of frequent flying. Eligibility was determined based on screening surveys. Career pilot radiation doses were calculated individually using airline flight profiles, personal flight history, and the CARI computer program. Translocation frequency was determined using fluorescence in situ hybridization. RESULTS: Blood samples for chromosome analysis were provided by 19 individuals. The mean number of metaphases counted per subject was 2802 in the pilots and 3000 in the controls. The mean number of translocations per cell (genome equivalent) was significantly higher among the pilots (mean +/- SE; 0.0031 +/- 0.0008) than among the controls (0.0010 +/- 0.0003) (p = 0.03, Mann-Whitney U test). However, within the 26 to 72 millisievert range encountered in this study, observed values among the pilots did not follow the dose-response pattern expected based on available models for chronic low dose radiation exposure. CONCLUSIONS: There was a statistically significant higher number of translocations per cell among pilots than among controls, although the expected dose-response relationship for radiation was not observed among the pilots.

[Gene therapy: new developments]. / Gentherapie: Neue Entwicklungen.

Gene therapy is based on the transfer and the expression of therapeutic genes in specific target cells. For the treatment of genetic diseases gene therapeutic approaches aim at replacement of the deficient gene or at the correction of the genetic defect. Malignant diseases and an increasing number of other acquired diseases are additional targets for gene therapeutic strategies. For gene therapy to become a potential future treatment option, safe and therapeutically efficient gene transfer into specific target cells is a central requirement. A variety of nonviral and viral vector systems have been developed. Nonviral vectors transfer genes are far less efficient than viral vectors, but they have advantages due to their low immunogenicity and their large size capacity for therapeutic DNA. To improve the function of nonviral vectors, the addition of viral functions such as receptor mediated uptake, enhanced endosomal release and nuclear translocation of DNA may finally lead to the development of an artificial virus. In contrast, natural viruses are already highly developed structures for the transfer of nucleic acids. Recombinant viruses can be used for efficient gene transfer. Retroviruses, adeno-associated viruses and lentiviruses are suitable for gene therapeutic approaches which are based on the permanent expression of therapeutic genes such as the correction of enzyme deficiencies or the manipulation of hematopoetic stem cells, as they are able to integrate nucleic acids into the cellular genome. In contrast, adenoviral vectors result in highly efficient, but transient gene expression and are therefore especially useful for the treatment of malignant tumors. Novel developments of viral vectors mainly aim at the reduction of immunogenicity, increase of capacity for therapeutic genes and at improved vector production. Viruses which replicate selectively in tumor cells leading to tumor cell lysis represent a novel generation of viral vectors, which can further be improved by the addition of therapeutic genes resulting in enhanced tumor toxicity.

Comparison of microscopic methods for detecting inflammation in expressed prostatic secretions.

PURPOSE: We evaluated microscopic methods of detecting inflammation in expressed prostatic secretions. MATERIALS AND METHODS: Methods of counting expressed prostatic secretion leukocytes were compared in 251 samples from 159 patients with chronic prostatitis/chronic pelvic pain syndrome, including traditional wet mounts, hemocytometer derived concentrations and expressed prostatic secretion smears stained with Gram's method or DiffQuick stain (Dade International, Inc., Miami, Florida). RESULTS: Of 159 initial patient evaluations 84 (53%) showed inflammation by hemocytometer concentration at 500 leukocytes per mm.3 or greater but only 37 (23%) were considered inflammation by the traditional wet mount method (p <0.001). Inflammation was identified in 149 of 251 specimens (59%) by hemocytometer but in only 82 (33%) by wet mount (p <0.001). When inflammation was defined as 1,000 leukocytes per mm.3 or greater the hemocytometer still identified significantly more patients (41%) and specimens (48%) with inflammation than the wet mount. The hemocytometer method had a substantially lower interassay and intra-assay coefficient of variation than the wet mount method. Polymorphonuclear neutrophils and macrophages were the most common cells observed on stained smears, which detected inflammation in 147 specimens (59%) by DiffQuick but in only 98 (39%) by Gram's method. CONCLUSIONS: Detecting inflammation in expressed prostatic secretions is method dependent. Significantly more cases of inflammation were detected by hemocytometer than by the traditional wet mount technique. Because the wet mount method also proved more variable than the hemocytometer and highly sensitive to volume, its use is not recommended. These findings support the adoption of hemocytometer and staining methods for accurate evaluation of expressed prostatic secretion inflammation in men with chronic prostatitis/chronic pelvic pain syndrome.

Immunotherapy directed against alpha-fetoprotein results in autoimmune liver disease during liver regeneration in mice.

BACKGROUND & AIMS: Priming immune responses against alpha-fetoprotein (AFP) highly expressed in the majority of hepatocellular carcinomas results in significant antitumoral T-cell responses. Liver regeneration in humans and mice, however, is also associated with increased AFP expression. Therefore, we evaluated the risk of AFP-directed immunotherapeutic approaches to induce autoimmunity against the regenerating liver. METHODS: Mice were immunized with DNA encoding mouse AFP. For induction of liver regeneration, partial hepatectomy was performed and mice were monitored by serial histopathologic examinations and measurements of serum ALT activities (U/L), and by determination of the kinetics of AFP-specific T-cell responses. RESULTS: Livers of AFP immune mice without partial hepatectomy were characterized by minor lymphocytic infiltrations without transaminase elevations. By contrast, a significant hepatocyte damage was observed in regenerating liver that correlated well with the number of AFP-specific CD8(+) T cells, the activity of liver regeneration, and the level of AFP synthesis. Autoimmune liver damage was mediated by CD4(+) T cell-dependent CD8(+) cytotoxic T lymphocytes. CONCLUSIONS: These results show that priming of T-cell responses against shared tumor-specific self antigens may be accompanied by induction of autoimmunity dependent on the level of expression of the self antigen and have important implications for the development of antitumoral vaccines targeted against antigens that are not strictly tumor-specific.

Adenovirus-mediated gene transfer to orthotopic hepatocellular carcinomas in athymic nude mice.

Gene therapy may become an option for the treatment of malignant tumors such as hepatocellular carcinoma (HCC), once safe and efficient vector systems have been established. Due to their stability in vivo, recombinant adenoviral vectors are promising vectors for gene delivery to HCC. To study the characteristics of gene delivery into HCCs by recombinant adenoviral vectors in vivo, we established an in situ HCC model in the livers of athymic nude mice by intrahepatic injection of human HCC cells. Recombinant adenovirus vectors expressing beta-galactosidase (Ad2CMV beta gal) were injected via the tail vein of mice bearing HCC or directly into intrahepatic tumors. Levels of beta-galactosidase expression in tumor tissue and surrounding normal liver were analyzed by histochemistry or for quantification by a chemiluminescence assay in tissue homogenates. Following tail vein injection, high levels of beta-galactosidase expression were found in the liver, but virtually no gene expression could be detected in the tumor tissue. In contrast, after direct injection of Ad2CMV beta gal into intrahepatic HCCs, high levels of beta-galactosidase expression were detected in the tumor tissue. However, single transduced hepatocytes scattered throughout the normal liver could also be identified. These results indicate that barriers such as the endothelial lining of the tumor vasculature impair the efficiency of adenoviral vectors for gene delivery into HCCs by intravenous administration, which can be overcome by direct injection into the tumor tissue. However, due to the observed transduction of disseminated hepatocytes following intratumoral administration, additional HCC-specific targeting to further enhance the safety of adenoviral vectors may be required.

Health among commercial airline pilots.

BACKGROUND: The airline pilot works within a complex exposure environment that may present physiological challenges to long-term health. METHODS: This study investigated self-reported disease outcomes among a large group of active and retired commercial airline pilots in the United States and Canada. A survey methodology was used, including the collection of historical information. RESULTS: Of 10,678 surveys mailed, 6609 were returned (6533 men, 63 women). Given the limitations of survey methodology, increased disease rates among pilots were suggested for melanoma, motor neuron disease, and cataracts. However, rates for other diseases were in general lower than those for the U.S. population. CONCLUSIONS: Further study has been initiated to verify and follow reported cases, to expand the study to a larger group, and to collect more in-depth information on flight histories, occupational exposures, and lifestyle factors.