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Biomedical subjects

T Wolff

Publications and source records attributed to T Wolff.

At least 19 recordsLinked to original sources
PubMed2002

NS1 protein of influenza A virus down-regulates apoptosis.

Wild-type (WT) influenza A/PR/8/34 virus and its variant lacking the NS1 gene (delNS1) have been compared for their ability to mediate apoptosis in cultured cells and chicken embryos. Cell morphology, fragmentation of chromatin DNA, and caspase-dependent cleavage of the viral NP protein have been used as markers for apoptosis. Another marker was caspase cleavage of the viral M2 protein, which was also found to occur in an apoptosis-specific manner. In interferon (IFN)-competent host systems, such as MDCK cells, chicken fibroblasts, and 7-day-old chicken embryos, delNS1 virus induced apoptosis more rapidly and more efficiently than WT virus. As a consequence, delNS1 virus was also more lethal for chicken embryos than WT virus. In IFN-deficient Vero cells, however, apoptosis was delayed and developed with similar intensity after infection with both viruses. Taken together, these data indicate that the IFN antagonistic NS1 protein of influenza A viruses has IFN-dependent antiapoptotic potential.

Amino Acid Sequence↗
PubMed2001

Dose dependent induction of DNA adducts, gene mutations, and cell proliferation by the antiandrogenic drug cyproterone acetate in rat liver.

1. CPA does not only induce the formation of DNA adducts but also of mutations in female rat liver. 2. The mutation frequency exhibited a characteristic time course. Within a period of 3 days post administration, a tremendous increase was noted, which remained at a high level until 2 weeks post exposure. Thereafter, most mutation-carrying cells were eliminated within a period of 2 weeks leaving a cell population remaining at a constant level for another 4 weeks. Thus, the length of the observation period post exposure, i. e. the manifestation time, seems to be a critical factor for the strength of the mutagenic response. The highest as observed between 1 and 2 weeks post exposure. Correspondingly, the dose response curve recorded 2 weeks post exposure showed a higher mutagenic response than the curve after 6 weeks of exposure recorded previously. 3. When CPA-induced mutations were recorded as a function of the dose, mutation frequencies at the lower dose range were found that did not differ from those of controls. The non-effective dose recorded 2 weeks post exposure was much lower than that recorded after 6 weeks of exposure indicating that it is a function of the manifestation time. Since DNA adducts were formed in high amounts at the non-effective doses, we assume that the mitogenic activity required for the conversion of DNA adducts into mutations was not sufficiently strong. The liver of adult animals exhibits a very low endogeneous proliferation rate, which is not likely to contribute significantly to the expression of mutations. We conclude that it is the mitogenic activity of CPA itself, which stimulates the expression of mutations.

Androgen Antagonists↗
PubMed2001

Community coalition building--contemporary practice and research: introduction.

Over the last 20 years, coalition building has become a prominent intervention employed in communities across America. Coalitions provide community psychologists and those in related fields with a chance to work with whole communities and to better understand how to create community change. As we reflect on the past two decades of community coalition building, there are many questions to be answered about this phenomenon. Why has there been such an upsurge in community coalition building activity? What is the impact of this activity? What have we as students of community learned? What are the questions that we need to be asking to improve the effectiveness of coalition building efforts and their evaluation? This set of articles will review the state of the art of community coalition building in both practice and research. The structure of the articles reflects a collaborative process, with multiple contributors from different disciplines, using a variety of formats. Because this is an evolving phenomenon where the questions asked are as important as the lessons learned, many of the major sections include dialogues with community experts from across the country and from multiple fields, including community psychology, public health, political science, public administration, and grassroots organizing.

Community Health Planning↗
PubMed2001

The future of community coalition building.

The future holds great promise for community coalitions as powerful interventions for community change. Community change is envisioned as: working with whole communities; increasing grassroots and civic engagement; promoting diversity, collaboration and, advocacy; increasing roles for professional technical assistance and evaluation; future changes in the role of government; and the building of healthy communities. Examples of these dimensions follow, as contributed by activists from a wide range of fields.

Community Health Planning↗
PubMed2001

A practitioner's guide to successful coalitions.

The highly complex practice of building successful community coalitions is explored. Key dimensions related to coalition success are identified and best practices are delineated. Nine dimensions are explored that are critical to coalition success: coalition readiness, intentionality, structure and organizational capacity, taking action, membership, leadership, dollars and resources, relationships, and technical assistance. Two coalition case studies follow the discussion of dimensions and illustrate the journey traveled to create successful community coalitions.

Benchmarking↗
PubMed2001

Influenza virus propagation is impaired by inhibition of the Raf/MEK/ERK signalling cascade.

Influenza A viruses are important worldwide pathogens in humans and different animal species. The functions of most of the ten different viral proteins of this negative-strand RNA virus have been well elucidated. However, little is known about the virus-induced intracellular signalling events that support viral replication. The Raf/MEK/ERK cascade is the prototype of mitogen-activated protein (MAP) kinase cascades and has an important role in cell growth, differentiation and survival. Investigation of the function of this pathway has been facilitated by the identification of specific inhibitors such as U0126, which blocks the cascade at the level of MAPK/ERK kinase (MEK). Here we show that infection of cells with influenza A virus leads to biphasic activation of the Raf/MEK/ERK cascade. Inhibition of Raf signalling results in nuclear retention of viral ribonucleoprotein complexes (RNPs), impaired function of the nuclear-export protein (NEP/NS2) and concomitant inhibition of virus production. Thus, signalling through the mitogenic cascade seems to be essential for virus production and RNP export from the nucleus during the viral life cycle.

Active Transport, Cell Nucleus↗
PubMed2000

[Mediastinal space-occupying lesion of uncertain histology: difficulties of differential diagnosis].

HISTORY AND ADMISSION FINDINGS: A 78-year-old woman had, for the previous 4 months, been aware of an easily moveable tumour in the jugular region. She had been feeling well except for general fatigue. Physical examination revealed a mass in the jugular region, a little over 2 cm in diameter. INVESTIGATIONS: Routine laboratory tests were normal, except for a raised erythrocyte sedimentation rate. After ultrasound, scintigraphy and computed tomography, a biopsy was performed. Histologically it was suspicious of a centroblastic centrocytic non-Hodgkin lymphoma, but further pathological investigation established the diagnosis of a malignant thymoma. TREATMENT AND COURSE: The tumour was resected and histological examination confirmed a malignant thymoma with infiltration of surrounding tissues. Radiotherapy was subsequently undertaken. CONCLUSION: Even though its evidence is low, a malignant thymoma or benign thymic tumour should be included in the differential diagnosis of a space-occupying mediastinal mass of unclear histology.

Aged↗
PubMed2000

Genotoxicity of urban air pollutants in the Czech Republic. Part II. DNA adduct formation in mammalian cells by extractable organic matter.

The study was aimed at determining the genotoxic potential of extractable organic matter (EOM) from ambient air particles PM10 (<10 micrometer) using mammalian cells in culture as test system. Air samples were collected in the course of summer and winter periods in two regions of the Czech Republic representing low and high levels of air pollution, the districts of industrial Teplice and rural Prachatice, respectively. EOM was fractionated by acid-base partitioning and silica gel column chromatography. Aliquots of fractions were incubated with cultured hepatocytes derived from male rats or Chinese hamster lung V79NH cells expressing nitroreductase activity but virtually no cytochrome P450 activity. DNA adduct levels were analyzed by 32P-postlabeling using butanol extraction for adduct enrichment. In hepatocytes, crude extracts caused the formation of substantial amounts of DNA reactive material being detectable in a broad diagonal radioactive zone (DRZ) in the chromatograms. Highest DNA adduct levels were found in the aromatic fractions and slightly polar fractions which contain most of the polycyclic aromatic hydrocarbons (PAH) and nitro-substituted PAH (nitro-PAH), respectively, comprising 75-90% of total adducts. This partitioning was independent of the sampling period and locality. In agreement with the higher average ambient air concentrations of PAH in the winter than the summer, 3-4-fold higher DNA adduct levels were detected in extracts sampled in the winter. Calculated on the basis of EOM/m(3), DNA adduct levels of samples collected in winter period were 10-fold higher than those collected in the summer period and 2-fold higher in Teplice than in Prachatice. Pretreatment of hepatocytes with 2,3,7,8-tetrachlorodibenzo-p-dioxin decreased DNA binding by 50-75%. In contrast to the findings in hepatocytes, in V79NH cells about 80% of the DNA adducts were caused by material in the slightly polar fractions appearing as distinct spots in the radiochromatograms. Seasonal variation of DNA adducts in V79NH cells was greater than variation between localities. Our results suggest that PAH as well as nitro-PAH are the main contributors to the genotoxicity of EOM derived from both industrial and rural areas. The results, furthermore, indicate that analysis of DNA adducts in mammalian cells in culture offers a suitable method for monitoring the genotoxicity of complex mixtures of environmental chemicals.

Air Pollutants↗
PubMed2000

Water-binding solid scintillators: synthesis, emission properties, and tests in 3H and 14C counting.

Spectral and time-resolved fluorescence properties as well as relative fluorescence quantum yields of carbodiimide derivatives of 2.5-diphenyloxazole (PPO) (prepared by H2S elimination from the corresponding thioureas), of some intermediates in the preparation, and of several other PPO derivatives were investigated in solution and in the solid state to test their suitability as solid scintillators. The carbodiimides reacted slowly with water under acidic conditions to yield ureas. These systems were compared with solid mixtures of other PPO derivatives with sodium sulfate as a drying agent, as chemically water-binding solid scintillators in 3H and 14C counting. Both the chemically and the absorptive water-binding scintillators proved capable of counting 3H and 14C decay, and open a way to the counting of aqueous samples by solid scintillators without a drying step.

Carbon Radioisotopes↗
PubMed2000

The influenza A virus M1 protein interacts with the cellular receptor of activated C kinase (RACK) 1 and can be phosphorylated by protein kinase C.

The M1 protein of influenza A virus has multiple regulatory functions during the infectious cycle, which include mediation of nuclear export of viral ribonucleoproteins, inhibition of viral transcription and a crucial role in virus assembly and budding. The only known modification of the M1 protein is by phosphorylation through yet-to-be-identified kinases. We postulated that at least some of the M1 functions are exerted or regulated through interactions with cellular components. In a screen for such cellular mediators, the protein receptor of the activated C-kinase (RACK 1) was identified by its interaction with the viral M1 protein in the yeast two hybrid system. The physical M1-RACK 1 interaction was confirmed in glutathione-S-transferase-based coprecipitation assays for the diverged M1 proteins of avian, swine and human influenza A virus strains. This conservation suggests that the M1-RACK 1 interaction is of general importance during influenza A virus infections. RACK 1 has previously been identified to specifically bind the activated form of protein kinase C (PKC) and is assumed to anchor the kinase at membranes in the vicinity of its substrates. Since the M1 protein becomes phosphorylated during influenza virus infection, we examined if PKC could catalyze the phosphate transfer. We demonstrate that virion-derived and recombinant M1 protein can indeed be efficiently phosphorylated by purified PKC. Moreover, in cell extracts, we detected M1 phosphorylation activity that was strongly reduced in the presence of the PKC-specific inhibitor compound GF109203X. These data suggest that PKC is the main M1-phosphorylating activity in the cell. Since both, the M1 protein and PKC have been shown to interact with RACK 1, we suggest that the M1-RACK 1 interaction is involved in M1 phosphorylation.

Animals↗
PubMed2000

Inspired oxygen fraction after cardiopulmonary bypass: effects on pulmonary function with regard to endothelin-1 concentrations and venous admixture.

Twenty consecutive patients undergoing cardiac surgery with cardiopulmonary bypass (CPB) were allocated at random to group 1 (n = 10, high inspired oxygen fraction (FIO2) after CPB), or group 2 (n = 10, moderate FIO2 after CPB). The effects of each FIO2 on arterial and mixed venous concentrations of endothelin-1 (ET-1) and its precursor, Big ET-1, were measured. Venous admixture was calculated to assess the efficiency of pulmonary gas exchange. Patients whose lungs had been ventilated with a FIO2 of 1.0 (exposure time 70 min) after weaning from the CPB machine had significantly greater arterial and mixed venous Big ET-1 concentrations and venous admixture than patients whose lungs were ventilated with a FIO2 of 0.35. In contrast, ET-1 concentrations in the two groups were not significantly different. A reduction of FIO2 from 1.0 to 0.6 reduced venous admixture without lowering endothelial peptide concentrations. On the first postoperative day all peptide concentrations were similar in the two groups, whereas venous admixture remained non-significantly higher in group 1. A short period of high FIO2 immediately after CPB increases endothelin concentrations and pulmonary venous admixture.

Aged↗
PubMed2000

A short leucine-rich sequence in the Borna disease virus p10 protein mediates association with the viral phospho- and nucleoproteins.

Borna disease virus (BDV) is unique among the non-segmented negative-strand RNA viruses of animals and man because it transcribes and replicates its genome in the nucleus of the infected cell. It has recently been discovered that BDV expresses a gene product of 87 amino acids, the p10 protein, from an open reading frame that overlaps with the gene encoding the viral p24 phosphoprotein. In addition, the p10 protein has been localized to intranuclear BDV-specific clusters containing viral antigens. Here, characterization of p10 interactions with the viral nucleoprotein p38/p39 and the p24 phosphoprotein is reported. Immunoaffinity chromatography demonstrated the presence of high-salt stable complexes of p10 containing the p24 and p38/p39 proteins in extracts of BDV-infected cells. Analyses in the yeast two-hybrid system and biochemical co-precipitation experiments suggested that the p10 protein binds directly to the p24 phosphoprotein and indirectly to the viral nucleoprotein. Mutational analysis demonstrated that a leucine-rich stretch of amino acids at positions 8-15 within the p10 protein is critical for interaction with p24. Furthermore, binding of p10 to the viral phosphoprotein was shown to be important for association with the BDV-specific intranuclear clusters that may represent the sites of virus replication and transcription in infected cells. These findings are discussed with respect to possible roles for the p10 protein in viral RNA synthesis or ribonucleoprotein transport.

Binding Sites↗
PubMed2000

Interdependence of hemagglutinin glycosylation and neuraminidase as regulators of influenza virus growth: a study by reverse genetics.

The hemagglutinin (HA) of fowl plague virus A/FPV/Rostock/34 (H7N1) carries two N-linked oligosaccharides attached to Asn123 and Asn149 in close vicinity to the receptor-binding pocket. In previous studies in which HA mutants lacking either one (mutants G1 and G2) or both (mutant G1,2) glycosylation sites had been expressed from a simian virus 40 vector, we showed that these glycans regulate receptor binding affinity (M. Ohuchi, R. Ohuchi, A. Feldmann, and H. D. Klenk, J. Virol. 71:8377-8384, 1997). We have now investigated the effect of these mutations on virus growth using recombinant viruses generated by an RNA polymerase I-based reverse genetics system. Two reassortants of influenza virus strain A/WSN/33 were used as helper viruses to obtain two series of HA mutant viruses differing only in the neuraminidase (NA). Studies using N1 NA viruses revealed that loss of the oligosaccharide from Asn149 (mutant G2) or loss of both oligosaccharides (mutant G1,2) has a pronounced effect on virus growth in MDCK cells. Growth of virus lacking both oligosaccharides from infected cells was retarded, and virus yields in the medium were decreased about 20-fold. Likewise, there was a reduction in plaque size that was distinct with G1,2 and less pronounced with G2. These effects could be attributed to a highly impaired release of mutant progeny viruses from host cells. In contrast, with recombinant viruses containing N2 NA, these restrictions were much less apparent. N1 recombinants showed lower neuraminidase activity than N2 recombinants, indicating that N2 NA is able to partly overrule the high-affinity binding of mutant HA to the receptor. These results demonstrate that N-glycans flanking the receptor-binding site of the HA molecule are potent regulators of influenza virus growth, with the glycan at Asn149 being dominant and that at Asn123 being less effective. In addition, we show here that HA and NA activities need to be highly balanced in order to allow productive influenza virus infection.

Animals↗
PubMed1999

Photorheological Effects in Aqueous Micellar Tetramethylammoniumhydrogen-2-dodecyl Malonate via Photodimerization of Acridizinium Bromide.

Aqueous solutions of tetramethylammoniumhydrogen-2-dodecyl malonate (TMHM) were investigated rheologically in the absence and presence of monomeric and photodimerized acridizinium bromide as solubilizates. Upon constant shearing a viscosity increase (accompanied by viscoelasticity) was observed in a certain range of TMHM concentration after an induction period. This indicates the formation of shear induced structures. Viscoelasticity was characterized via stress relaxation experiments revealing a single relaxation process with time constants of 0.2-0.3 s mostly. At certain shear rates a temporal oscillation of viscosity was observed. In the presence of acridizinium bromide the elasticity was reduced and the TMHM concentration range of viscoelastic flow was narrowed while relaxation time constants remained constant. Photodimerization of acridizinium bromide (performed in situ) reverted the effects induced by the monomers. Copyright 1999 Academic Press.

Journal Article↗
PubMed1999

A fatal relationship--influenza virus interactions with the host cell.

Influenza A viruses are important worldwide pathogens for humans and different animal species. The infectious agent is the prototype of the orthomyxoviridae which are characterized by a segmented negative strand RNA genome that is replicated in the nucleus of the infected cell. The genome has a combined coding capacity of about 13 kb and contains the genetic information for ten viral proteins. Despite this relatively small coding capacity--large DNA viruses like herpes or poxviruses express about 150-200 gene products--influenza A viruses are able to successfully infect and multiply in a wide range of mammalian and avian species. It is therefore not surprising that influenza A viruses extensively use and manipulate host cell functions. This includes multiple interactions of viral proteins with cellular proteins. In recent years an increasing amount of information about the identity of the cellular factors that are involved in viral transcription and replication, intracellular trafficking of viral components and assembly of the virus particle has accumulated. This article aims to review recent developments in this field with a focus on cellular factors and processes which are activated by the virus to either support viral replication or to counteract host-cell defense mechanisms.

Animals↗
PubMed1998

DNA adduct formation in mammalian cell cultures by polycyclic aromatic hydrocarbons (PAH) and nitro-PAH in coke oven emission extract.

Mammalian cells in culture were used to study the genotoxic potential of coke oven emissions constituting a complex mixture of chemicals. For this purpose, particle extracts and some polycyclic aromatic and nitroaromatic hydrocarbons (PAH and nitro-PAH) occurring in these mixtures were assayed for DNA adduct formation using the -postlabeling technique. In primary cultures of rat hepatocytes, benzo[a]pyrene (B[a]P), benz[a]anthracene (B[a]A) and benzo[b]fluoranthene (B[k]F) caused DNA adduct levels in the range of 1 adduct/108 nucleotides. 4-Nitropyrene (4-NP), 6-nitrochrysene (6-NC), 3-nitrofluoranthene (3-NF) caused DNA adduct levels that were by one to two orders of magnitude higher. The crude particle extract and its fractions differing in acidity and polarity induced the formation of DNA reactive material within diagonal radioactive zones (DRZ) on the autoradiograms. On a weight base, the neutral aromatic fraction contributed by more than 80% to the total adduct level in hepatocytes. To examine whether the PAH- and nitro-PAH-DNA derived adducts can be further differentiated, hepatocyte cultures were preincubated with 2,3,7,8-tetrachloro-p-dioxin (TCDD) to induce the activity of cytochrome P450 1A1. TCDD pretreatment strongly increased the levels of PAH-DNA adducts, whereas, the levels of nitro-PAH adducts were markedly decreased. NCI-H322 cells, a human lung tumor cell line derived from Clara cells, exhibited PAH-DNA adduct levels between 10 and 100, and nitro-PAH-DNA adducts at levels between 0.2 to about 30 adducts per 108 nucleotides, respectively. In contrast to hepatocytes, incubations with extractable organic matter (EOM) and the neutral aromatic EOM fraction displayed several distinct spots in the chromatograms of NCI-H322 cells. The major spot was assigned by cochromatography to be identical with the major DNA adduct formed by incubation with B[a]P alone. In V79NH cells, a Chinese hamster lung cell line expressing nitro-PAH activating enzymes, but virtually no cytochrome P450 activity, PAH-derived DNA adducts were not detectable. Nitro-PAH-derived DNA adducts, however, were formed at levels between 10 and 300 adducts/108 nucleotides. The slightly and the moderately polar EOM fraction caused the formation of distinct adduct spots suggesting the occurrence of nitro-PAH in these fractions. GC/MS analyses revealed the presence of twelve PAH in the aromatic fraction, at a total amount of about 10% (w/w), and of four nitro-PAH in the slightly polar and the acidic fraction amounting to about 0.2% (w/w). In conclusion, our results indicate that PAH and nitro-PAH contribute to the genotoxicity of coke oven emissions. Using DNA adduct analysis in rat hepatocytes (+/-pretreatment with TCDD) and in NCI-H322 and in V79NH cells offers a promising approach to determine the genotoxic activity of PAH and nitro-PAH in any complex environmental samples.

Air Pollutants↗