Krankheiten als Folge der Selbstverkennung und Selbstzersetzung von Geweben

Ohne ZusammenfassungVortrag gehalten vor der 105. Versammlung der Gesellschaft Deutscher Naturforscher und Ärzte, Heidelberg, Oktober 1968.     

Monkeypox virus detection in rodents using real-time 3'-minor groove binder TaqMan assays on the Roche LightCycler

During the summer of 2003, an outbreak of human monkeypox occurred in the Midwest region of the United States. In all, 52 rodents suspected of being infected with monkeypox virus were collected from an exotic pet dealer and from private homes. The rodents were euthanized and submitted for testing to the United States Army Medical Research Institute of Infectious Diseases by the Galesburg Animal Disease Laboratory, Illinois Department of Agriculture. The rodent tissue samples were appropriately processed and then tested by using an integrated approach involving real-time polymerase chain reaction (PCR) assays, an antigen-detection immunoassay, and virus culture. We designed and extensively tested two specific real-time PCR assays for rapidly detecting monkeypox virus DNA using the Vaccinia virus F3L and N3R genes as targets. The assays were validated against panels of orthopox viral and miscellaneous bacterial DNAs. A pan-orthopox electrochemiluminescence (ECL) assay was used to further confirm the presence of Orthopoxvirus infection of the rodents. Seven of 12 (58%) animals (seven of 52 (15%) of all animals) tested positive in both monkeypox-specific PCR assays and two additional pan-orthopox PCR assays (in at least one tissue). The ECL results showed varying degrees of agreement with PCR. One hamster and three gerbils were positive by both PCR and ECL for all tissues tested. In addition, we attempted to verify the presence of monkeypox virus by culture on multiple cell lines, by immunohistology, and by electron microscopy, with negative results. Sequencing the PCR products from the samples indicated 100% identity with monkeypox virus strain Zaire-96-I-16 (a human isolate from the Congo). These real-time PCR and ECL assays represent a significant addition to the battery of tests for the detection of various orthopoxviruses. In light of the recent monkeypox virus transmissions, early detection of the virus is crucial for both natural outbreaks and potential acts of bioterrorism.     

Präkanzerosen des Urothels

Zusammenfassung In der Fluoreszenzdiagnostik [Lichtanregung nach Gabe des physiologischen Hämmetaboliten 5-Aminolävulinsäure (ALA)] erscheinen Präneoplasien und Neoplasien rot fluoreszierend, was wahrscheinlich auf eine tumorzellselektive Anreicherung des intrazellulär aus ALA gebildeten Metaboliten Protoporphyrin IX zurückzuführen ist. Die mit dieser Methode mögliche frühe Detektion von Tumoren und Präneoplasien erlaubt deren genetische Analyse und damit Vergleiche von Aussagen der Fluoreszenz-in-situ-Hybridisierung und Loss-of-heterozygosity-Analyse hinsichtlich Deletionsnachweis und Gensequenzierung. Neue Daten zu Deletionen und Chromosomenaberrationen sowie eine etwa 30%ige Oligoklonalität von Tumoren werden dargestellt. Die tumorselektive Fluoreszenz scheint biochemisch durch Unterschiede im Hämmetabolismus bedingt zu sein. Durch Gen- und Proteinexpressionsanalysen können evtl. weitere, mit dem Hämmetabolismus assoziierte, tumorspezifische Moleküle identifiziert werden.Die geschilderten Arbeiten erfolgten im Rahmen von Unterstützungen durch die Deutsche Krebshilfe 10-1096-Ha und 70.2200-Ba sowie Industriekooperationen von Medac GmbH und metaGen Pharmaceuticals.     

Buchbesprechungen

Ohne Zusammenfassung     

Histopathologic analysis of atypical lesions in image-guided core breast biopsies.

Appropriate follow-up of patients with needle core breast biopsies (NCBB) showing atypical hyperplasia remains unclear because previous studies show that subsequent open biopsies in variable proportions of these patients reveal ductal carcinoma in situ (DCIS) or even invasive carcinoma, indicating significant sampling artifact. NCBB with diagnoses of atypia were morphologically classified into groups as follows: I, ALH (n = 24); II, ADH with minimal cytologic atypism (n = 90); III, atypia, other (9 columnar, 2 apocrine, 11 atypical papillary); IV, severe ADH/borderline DCIS (n = 31). Mammographic and histologic features, including the number of foci of atypia in the NCBB and the calcification span, were then correlated with presence of DCIS or invasive tumor in subsequent open excisions. Open excisional biopsies showed more severe lesions in 12% of Group I-III cases (8% in Group I, 9% in Group II, and 27% in Group III), of which 15 were DCIS and one was an invasive tubular carcinoma (0.3 cm). Of the DCIS, 60% (n = 9) were < or =5 mm, and 13 of 15 (87%) were low grade. The NCBB cavity was immediately adjacent to the more severe lesions in 88% (n = 14) of cases, in keeping with sampling error. The subset showing severe ADH with borderline nuclear features in contrast was associated with a high likelihood (63%) of DCIS in follow-up excisions. NCBB with atypical papillary features also showed a high frequency of DCIS (4/11, 36%) in subsequent open excisions. Other factors associated with more severe lesions on open biopsy included the number of atypical foci in the NCBB (>4, P <.05) and the mammographic calcification span (>2.0 cm, P <.0001). Atypical lesions diagnosed in NCBB samples are radiographically and morphologically heterogeneous, accounting for the variable frequency of DCIS or invasive neoplasm identified in subsequent open excisions, which are usually focal, low grade, and a consequence of sampling artifact (i.e., adjacent to the NCBB cavity). DCIS is more likely if microcalcifications are mammographically extensive or if atypia is multifocal or is associated with borderline cytologic features.     

Detection of DNA damage after hyperbaric oxygen (HBO) therapy

Hyperbaric oxygen HBO therapy is successfully used for the treatmentof a variety of conditions. However, exposure to high concentrationsof oxygen is known to induce damage to cells, possibly due toan increased oxygen radical production. As reactive oxygen speciesalso cause DNA damage, we investigated the DNA-damaging effectof HBO with the alkaline version of the single cell gel testcomet assay. Oxidative DNA base modifications were determinedby converting oxidized DNA bases to strand breaks using bacterialformamidopyrimidine-DNA glycosylase FPG, a DNA repair enzyme,which specifically nicks DNA at sites of 8-oxo-guanines andformamidopyrimidines. HBO treatment under therapeutic conditionsclearly and reproducibly induced DNA damage in leukocytes ofall test subjects investigated. Increased DNA damage was foundimmediately at the end of the treatment, while 24 h later, noeffect was found. Using FPG protein we detected significantoxidative base damage after HBO treatment DNA damage was detectedonly after the first treatment and not after further treatmentsunder the same conditions, indicating an increase in antioxidantdefences. DNA damage did not occur when the HBO treatment wasstarted with a reduced treatment time which was then increasedstepwise. ³To whom correspondence should be addressed     

Phase I testing of a malaria vaccine composed of hepatitis B virus core particles expressing Plasmodium falciparum circumsporozoite epitopes

We report the first phase I trial to assess the safety and immunogenicity of a malaria vaccine candidate, ICC-1132 (Malarivax), composed of a modified hepatitis B virus core protein (HBc) containing minimal epitopes of the Plasmodium falciparum circumsporozoite (CS) protein. When expressed in Escherichia coli, the recombinant ICC-1132 protein forms virus-like particles that were found to be highly immunogenic in preclinical studies of mice and monkeys. Twenty healthy adult volunteers received a 20- or a 50-microg dose of alum-adsorbed ICC-1132 administered intramuscularly at 0, 2, and 6 months. The majority of volunteers in the group receiving the 50-microg dose developed antibodies to CS repeats as well as to HBc. Malaria-specific T cells that secreted gamma interferon were also detected after a single immunization with ICC-1132-alum. These studies support ICC-1132 as a promising malaria vaccine candidate for further clinical testing using more-potent adjuvant formulations and confirm the potential of modified HBc virus-like particles as a delivery platform for vaccines against other human pathogens.     

Multilineage potential of homozygous stem cells derived from metaphase II oocytes

Human stem cells derived from human fertilized oocytes, fetal primordial germ cells, umbilical cord blood, and adult tissues provide potential cell-based therapies for repair of degenerating or damaged tissues. However, the diversity of major histocompatibility complex (MHC) antigens in the general population and the resultant risk of immune-mediated rejection complicates the allogenic use of established stem cells. We assessed an alternative approach, employing chemical activation of nonfertilized metaphase II oocytes for producing stem cells homozygous for MHC. By using F1 hybrid mice (H-2-B/D), we established stem cell lines homozygous for H-2-B and H-2-D, respectively. The undifferentiated cells retained a normal karyotype, expressed stage-specific embryonic antigen-1 and Oct4, and were positive for alkaline phosphatase and telomerase. Teratomatous growth of these cells displayed the development of a variety of tissue types encompassing all three germ layers. In addition, these cells demonstrated the potential for in vitro differentiation into endoderm, neuronal, and hematopoietic lineages. We also evaluated this homozygous stem cell approach in human tissue. Five unfertilized blastocysts were derived from a total of 25 human oocytes, and cells from one of the five hatched blastocysts proliferated and survived beyond two passages. Our studies demonstrate a plausible "homozygous stem cell" approach for deriving pluripotent stem cells that can overcome the immune-mediated rejection response common in allotransplantation, while decreasing the ethical concerns surrounding human embryonic stem cell research.