PubMed Health⌕ Search

Biomedical subjects

H W Doerr

Publications and source records attributed to H W Doerr.

At least 163 records · Page 9Linked to original sources
PubMed1992

Aphidicolin selectively kills neuroblastoma cells in vitro.

Aphidicolin is a tetracyclic diterpene antibiotic which is known to inhibit the growth of eucaryotic cells by reversible binding to DNA polymerase alpha without significant effect on cell viability in most common human cell lines. We observed that aphidicolin at a concentration of 5 x 10(-7) M kills all cells of four human neuroblastoma cell lines. In contrast, viability of normal human embryonal cells and of human continuous cell lines including HeLa, H9, A549 and Caco-2 was influenced only moderately by aphidicolin. In addition, neuroblastoma cells were killed after treatment with 5 x 10(-7) M aphidicolin in cocultures with normal embryonal cells which continued to proliferate after removal of aphidicolin. These results show that aphidicolin provides an agent which selectively kills neuroblastoma cells in vitro.

Antineoplastic Agents↗
PubMed1992

Human cytomegalovirus (HCMV)-specific immunoglobulin E as a serologic marker for HCMV infection in immunocompromised patients.

An antibody capture assay using an enzyme-linked human cytomegalovirus (HCMV) antigen for the detection of specific immunoglobulin E (IgE) was established. IgG, M, and E responses to HCMV were studied in 497 sera obtained from 44 renal transplant recipients and 51 acquired immunodeficiency syndrome (AIDS) patients. The results were compared with those obtained from 58 HCMV-seropositive healthy individuals. HCMV-specific IgE was detected in 11 (91.7%) renal transplant recipients with primary HCMV infection. In contrast, antibodies of the IgG and IgM classes were detected in only 6 (50.0%) of these patients. Specific IgE was detected in 10 (90.9%) out of 11 renal allograft recipients suffering from secondary HCMV infection. Significant IgG titer rises and IgM were detected in 2 (18.2%) and 6 (54.6%) of these patients, respectively. IgG titer rises and IgM and IgE antibodies were seen in 5 (12.2%), 1 (2.4%) and 18 (43.9%) AIDS patients respectively. All healthy immunocompetent HCMV-seropositive individuals were tested IgE negative. The results obtained in our study indicate that IgE against HCMV is a more reliable serologic marker for primary and secondary HCMV infection than IgM in immunocompromised individuals, especially in organ transplant recipients, since it is not affected by the prophylactic application of HCMV hyperimmune globulin preparations.

Acquired Immunodeficiency Syndrome↗
PubMed1992

Comparison of shell viral culture and serology for the diagnosis of human cytomegalovirus infection in neonates and immunocompromised subjects.

The present retrospective study compares the laboratory diagnosis of cytomegalic inclusion disease (CID) by the use of "shell vial culture" [i.e., immunoperoxidase staining of human cytomegalovirus (HCMV) early antigen in human fibroblasts 24 h postinoculation] to the results of serology (i.e. immunoglobulins IgG, IgM, and IgA HCMV antibody testing) in 21 infants with congenital or postnatally acquired HCMV infection, 5 patients with lymphoproliferative disorders, 35 human immunodeficiency virus (HIV)-seropositive patients who met the Centers for Disease Control (CDC) criteria for stages IVA and IVB of HIV infection, and 115 patients suffering from the acquired immunodeficiency syndrome, AIDS (stages IVC-IVE according to CDC criteria). HCMV infection was diagnosed by means of the shell vial culture inoculated with patient samples (e.g., urine, bronchoalveolar lavage, induced sputum, etc.) and serology in 163 (92.6%) and 65 (36.9%) patients, respectively. Viral shedding was detected by shell vial culture in 100% of the neonates, 80% of the patients suffering from lymphoproliferative disorders, 100% of the AIDS related complex (ARC) and 89.6% of the AIDS patients. In contrast, serologic testing for HCMV-specific antibodies was positive in only 28.6%, 42.9%, and 34.8% of the neonates, ARC, and AIDS patients, respectively. In lymphoma patients, serologic testing gave identical results (80%) to the shell vial culture technique. With the use of the shell vial procedure, active HCMV infection in immunocompromised subjects and neonates can be recognized more reliably than by serologic testing. Nevertheless, in a low percentage of patients (7.4%), virus isolation by the shell vial culture may fail to detect HCMV infection.

Acquired Immunodeficiency Syndrome↗
PubMed1992

Increased HIV-1 production in chronically infected H9 cells grown in protein-free medium.

Human T cell line H9 was established in a protein-free 1:1 mixture of Ham's F-12 and IMDM. After 230 passages (3 years) in protein-free medium, the cells designated H9-PF were infected with HIV-1. The infectivity titers of HIV-1 in cell culture medium were monitored by determining the median tissue culture infectious doses (TCID50). Additionally, the production of viral antigen in cells was measured by an immunoenzymatical alkaline phosphatase anti-alkaline phosphatase (APAAP) method using a monoclonal antibody against HIV-1-p24 antigen. In acutely infected H9-PF and H9 cultures similar TCID50 values and percentage of cells positive for p24 antigen were found. In contrast, both TCID50 values and percentage of cells positive for p24 antigen were by far greater in chronically infected H9-PF than in H9 cultures.

Cell Line↗
PubMed1992

Improvement of rotavirus isolation in the cell culture by immune peroxidase staining.

Peroxidase-labeled monoclonal antibody against rotavirus group-specific antigen (inner capsid) was used for the detection of rotavirus by immunoperoxidase staining (IPS) in trypsin-free MA104 cells within 18 h post-inoculation with clinical specimens. One hundred and twenty-one fecal samples from children with acute gastroenteritis were evaluated by IPS, conventional virus isolation in cell culture and a commercially available group A-antigen ELISA (Rotazyme II, Abbott Laboratories). Fifty-eight (47.9%) stool samples were found positive by IPS. In contrast, rotavirus was isolated from only 4 (3.3%) fecal specimens by conventional cell culture (i.e. demonstration of a cytopathogenic effect). A total of 93 (76.9%) samples were positive by ELISA. IPS permits rapid detection of rotavirus infections and detects shedding of infectious virus. The method should be useful for the investigation of nosocomial spread of rotavirus infection in hospitals, contamination of environmental surfaces and desinfectants.

Cells, Cultured↗
PubMed1992

Polyvinyl formal surface promotes continuous growth of Vero cells in protein-free medium.

The effect of polyvinyl formal (PVF) culture surface on the growth of 10 mammalian continuous cell lines, including Swiss 3T6, NCTC clone 929 L, BHK-21 clone 13, CHO-K1, PK 15, A 431, HeLa, MDCK, LLC-MK2 and Vero in protein-free 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 supplemented with trace elements and L-ascorbic acid 2-phosphate, was investigated. Most of the cell lines showed only some initial proliferation on PVF similar to the polystyrene (PS) surface of commercially available culture flasks. In contrast, proliferation of monkey kidney cell line Vero was by far greater on PVF than on PS or poly-D-lysine treated culture surface. In addition, Vero cells on PVF could be subcultured in the protein-free medium without any significant decrease of growth rate in successive passages. These results showed that PVF provides a culture surface which selectively promotes continuous growth of Vero cells in protein-free, chemically defined medium.

Animals↗
PubMed1992

In-vitro activity of cefpirome against isolates from patients with urinary tract, lower respiratory tract and wound infections.

The in-vitro activity of cefpirome, a new injectable cephalosporin, was compared with that of several other antibiotics against bacterial isolates from hospitalized patients with lower respiratory tract infections, urinary tract infections and wound infections. Minimum inhibitory concentrations were determined for 874 strains against 16 antibiotics using a microtitration technique. Cefpirome showed a very broad spectrum of activity against most pathogens tested. The spectrum included organisms such as Staphylococcus spp., enterococci, Enterobacter spp., and Pseudomonas spp. which are frequently resistant to third generation cephalosporins.

Anti-Bacterial Agents↗
PubMed1992

Multicenter evaluation of the novel ABN Western blot (immunoblot) system in comparison with an enzyme-linked immunosorbent assay and a different Western blot.

A new, modular Western blot (immunoblot) system for human immunodeficiency virus (HIV) antibodies (ABN WesPage; Wellcome) was compared with enzyme immunoassays (Wellcome, Behringwerke, and Abbott) and with a U.S. Food and Drug Administration (FDA)-licensed Western blot (DuPont) in a multicenter study. A total of 649 serum samples from HIV patients at different stages of the disease, as well as from high-risk patients, from patients with conditions unrelated to AIDS, and from healthy blood donors, were used in the evaluation along with nine seroconversion panels. For evaluation of Western blot reactivity, both Centers for Disease Control (CDC) and FDA criteria were used. With the DuPont Western blot as the reference assay, the overall sensitivity and specificity of the ABN WesPage were 100 and 99.1%, respectively, when indeterminate results were not taken into account and when both tests were interpreted in accordance with CDC criteria. The DuPont Western blot detected significantly more antibodies to pol and gag gene products than the ABN WesPage. The ABN WesPage showed a higher positive rate of detection of viral envelope band gp160. When both Western blots were interpreted in accordance with CDC criteria, the ABN WesPage and the DuPont Western blot yielded 9.3 and 10.4% indeterminate results, respectively. When the DuPont Western blot was interpreted in accordance with the manufacturer's instructions (FDA criteria), 25.7% of the samples tested were regarded as indeterminate. The choice of interpretation criteria is of paramount importance for the evaluation of HIV Western blot patterns.

AIDS Serodiagnosis↗
PubMed1992

[The detection of infectious rotaviruses by cell culture technique: use and evaluation of the immunoperoxidase assay].

In the present study, virus isolation was attempted in one hundred and twenty-one fecal samples of children suffering from acute gastroenteritis. Virus isolation was performed either conventionally by examination of cytopathogenic effect (CPE) or by immunoperoxidase staining (IPS) of rotavirus group specific antigen (inner capsid) in trypsin free MA104 cells within 18 h. Applying the conventional technique, rotavirus was isolated in only 4 (3.3%) fecal specimens. In contrast, IPS detected infectious virus in 49 (40.5%) stool samples. The specificity of IPS was confirmed by the results obtained with an antigen detection ELISA ("Rotazyme", Abbott, Wiesbaden) and gel electrophoresis of rotaviral RNA (electrophoretyping). ELISA and RNA gel electrophoresis detected rotavirus in 66 (54.4%) and 56 (46.3%) stool samples respectively. IPS enables rapid diagnosis of rotavirus infections in cell cultures applied to detect infectious viral particles in order to be used in the investigation of nosocomial outbreaks, material- and surface contamination and evaluation of disinfectants.

Acute Disease↗
PubMed1992

Effect of aciclovir on the replication of herpes simplex virus type 1 in MA-104 cell line resistant to aciclovir.

A culture of monkey kidney cells (MA-104) resistant to aciclovir (CAS 59277-89-3) was established. The resistant cells were continuously grown in medium containing 200 mumol/l aciclovir whereas normal cells died in such medium. The doses of aciclovir required for 50 and 90% reduction in virus yield were for several Herpes simplex virus Type 1 clinical isolates up to 5-fold higher in resistant than in normal MA-104 cells; in contrast, doses required for 99% reduction in virus yield could be as much as 50-fold higher in the resistant cells. Virus yields developed in the presence of 30 mumol aciclovir in culture medium were at least 10(3)-fold higher in resistant than in normal MA-104 cells. The results showed that the antiviral effect of aciclovir may be significantly decreased in the cell line which developed resistance to aciclovir.

Acyclovir↗
PubMed1991

[The status of immunity against poliomyelitis. The 5th cooperative study of the German Union to Control Viral Diseases, Inc].

Neutralizing antibodies against poliovirus types 1, 2 and 3 were determined in the serum samples, supplied by 11 virological institutes, from 3,341 subjects of different ages living in the western part of the Federal Republic of Germany. The 3 poliovirus types were found in all age groups, with a seroprevalence of at least 80%. Although antibodies against the 3 types of poliovirus in the age-groups up to 30 years were demonstrated more frequently than in the previous years in which samples had been examined (1969, 1972, 1978 and 1983), 33% of subjects over 40 years (this age-group had not previously been tested) did not have complete immunity against all 3 poliovirus types. For this reason immunization against poliomyelitis should continue to be recommended when indicated, especially before travels to regions where the disease is endemic.

Age Factors↗
PubMed1991

IgG subclass-specific antibodies to human cytomegalovirus (HCMV)-induced early antigens.

We investigated the IgG subclass reactivity pattern to early antigens (EA) of human cytomegalovirus (HCMV) in 217 EA-antibody-positive sera from immunocompetent, healthy persons, renal transplant recipients and AIDS patients by immunoblotting. All IgG subclasses are involved in the IgG immune response to HCMV EA. IgG 1 was the major subclass reacting with HCMV EA (with a molecular mass ranging between 23- and 79-kDa) and was present in all sera irrespective of origin. Antibody responses of IgG isotypes 2, 3 and 4 were observed with lower frequency and reactivity, whereas IgG 3 was detectable more frequently and reacted slightly stronger than IgG 2 and 4. The IgG 1 reactivity pattern was similar to that seen with total IgG. In contrast to total IgG and IgG 1, the reactivity of the sub-classes 2, 3 and 4 was not equally distributed among the early polypeptides, but was mainly directed to some of them (79-, 70-, 66-, 43- and 38-kDa). On average primary infections seem to induce a stronger IgG 3 response to the 79-, 70- and 38-kDa proteins than reactivated infections and an increased IgG 1 to the 79-, 70-, 59-, 56- and 50-kDa proteins appeared to be associated with severe disease. Noteworthy was the significantly lower prevalence of IgG 1 antibodies to the 70-kDa protein in human immunodeficiency virus (HIV)-infected individuals when compared to renal transplant recipients and immunocompetent, healthy individuals. Since the IgG 1 immune reaction to this protein occurred five to seven times more frequently in healthy persons and renal transplant recipients, the decreased formation of IgG 1 antibodies to the 70-kDa protein might be a characteristic feature of HIV infection.

Acquired Immunodeficiency Syndrome↗
PubMed1991

Quality control testing of surfaces for mammalian cell culture using cells propagated in a protein-free medium.

Mouse NCTC clone 929 L (L-929) cells were propagated continuously for 3 years as monolayers in a protein-free chemically-defined medium. These cells, designated L-929-WS, were used for quality control testing of the surfaces of commercially available cell culture plastic flasks. Differences in attachment and saturation density of L-929-WS cells in a protein-free culture medium were taken to define various levels of quality of the culture vessels tested. The rate of attachment and growth of L-929-WS cells on a surface of a given quality correlated directly with that of human embryonal fibroblasts and embryonal epithelial cells grown in a serum-free medium supplemented with growth factors and hormones. L-929-WS cells propagated continuously in a protein-free medium provide a simple and sensitive assay system for more general quality control testing of surfaces used for the culture of monolayer cells.

Animals↗
PubMed1991

2',3'-Dideoxycytidine preferentially inhibits in vitro growth of granulocyte-macrophage colony-forming cells from patients with chronic myeloid leukemia.

The effect of three nucleoside analogs, including 2-methyl-2'-deoxyadenosine (MDA), 5-amino-2'-deoxyuridine (ADU) and 2',3'-dideoxycytidine (DDC) on colony formation from unfractionated human bone marrow obtained from volunteers expressing parameters typical for normal cells (NBM) and from patients with the diagnosis of chronic myeloid leukemia (CMLBM) was observed. For the clonal growth of granulocyte-macrophage colony-forming cells (GM-CFC), a semisolid fibrin clot culture medium supplemented with 20% fetal bovine serum and 10% human placental conditioned medium was used. DDC has been shown to be at least a 10-fold more potent inhibitor for the growth of GM-CFC from CMLBM than from NBM. On the other hand, the effect of MDA and ADU on CMLBM did not differ markedly from the effect on NBM. These results suggest that DDC inhibits preferentially progenitor cells from CMLBM.

Bone Marrow↗
PubMed1991

[Atypical pathogen spectrum in community-acquired infections of the lower respiratory tract in childhood].

Over a period of 14 months, we obtained nasal and throat swabs for virological examinations from patients, admitted to hospital with suspected pneumonia. Fifty-five cases of perihilar bronchitis and 103 cases of pneumonia were diagnosed based on radiological criteria. In 36% (n = 57) of the children, a pathogen could be determined by antigen ELISA. We found the following spectrum: Respiratory syncytial virus: 42%; Influenza A: 8%; Influenza B: 10%; Parainfluenza 1: 3%; Parainfluenza 2: 10%; Parainfluenza 3: 2%; Chlamydia: 17%; and Mycoplasma pneumoniae (serology only): 7%. In a control group of 59 healthy children, only one child showed a pathogen. In the past, viruses and Chlamydia have been underestimated as causes of lower respiratory tract infections. Antigen determination by ELISA has proved to be a reliable and rapid method for their detection.

Adolescent↗