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Biomedical subjects

D Neumann-Haefelin

Publications and source records attributed to D Neumann-Haefelin.

At least 19 recordsLinked to original sources
PubMed1992

Genomic organization and expression of simian foamy virus type 3 (SFV-3).

The complete nucleotide sequence of simian foamy virus type 3 (SFV-3) strain LK-3, isolated from an African green monkey, was determined. In addition to translation frames representing the gag, pol, and env genes, two open reading frames are located in the region between the env gene and the 3' long terminal repeat (LTR). Both SFV-3 and SFV-1 encode two open reading frames between env and the 3' LTR, whereas HFV encodes three open reading frames in this region. Northern blot analysis of cell cultures infected with SFV-3 revealed subgenomic RNAs for these open reading frames. The protease of SFV-3 is encoded by the pol gene in contrast to HFV which encodes the protease in the gag gene. Notably, the pol gene of SFV-3 in the +1 translational frame relative to the gag gene; this observation is in agreement with SFV-1, but differs for HFV and all other retrovirus genomes reported. Thus, gag-pol precursors of the SFVs appear to be expressed by a +1 frameshift. Nucleotide and deduced amino acid alignments of SFV-3, SFV-1, and HFV revealed an unexpected homology pattern; highest homologies are observed in the pol and env genes but low homologies are noted in the gag genes and the additional open reading frames. Analysis of phylogenetic trees confirms the classification of foamy viruses as a subfamily of retroviruses, distinct from the lentiviruses and oncoviruses.

Amino Acid Sequence↗
PubMed1992

[Neurologic complications of Q-fever (Coxiella burnetii)].

We report on fifteen patients with a typical syndrome of Q-fever. Four patients (= 27%) showed signs of nervous system involvement. One patient developed a meningitic syndrome, another patient had a passing psychosis. In two patients, severe cerebellar deficits were predominant. It is pointed out that these two patients were under lithium therapy because of manic-depressive illness. A synergetic neurotoxic effect of cociella burneti infection and lithium is discussed as a possible cause of the severe cerebellar pattern of symptoms.

Adult↗
PubMed1992

Diagnosis and management of the acute retinal necrosis syndrome.

The acute retinal necrosis (ARN) syndrome is an increasingly occurring entity characterized by the triad of acute confluent peripheral retinitis with papillitis and anterior-chamber uveitis. We present case reports on four patients (age, 12-65 years) with an ARN syndrome caused by herpes simplex or varicella zoster virus and discuss diagnostic and therapeutic modalities. Immediate antiviral therapy in three patients exhibiting the typical clinical features reduced the intraocular inflammation. However, due to proliferative vitreoretinopathy with peripheral retinal necrosis, vitrectomy with encircling band and silicone oil instillation was necessary in all patients. The suspected diagnosis of an ARN syndrome induced by herpes simplex virus (HSV) was confirmed in one case during the early stage of the disease by the detection of increased levels of HSV-IgA in the vitreous and in another case by the measurement of increased titers of HSV-IgG in the vitreous. For the first time, we found intraocular HSV DNA sequences using the polymerase chain reaction (PCR) in one of these patients. In a fourth patient intraocular varicella zoster virus (VZV) infection was confirmed by the detection of elevated VZV-IgA levels and by positive PCR in the intraocular fluids. Two patients who were diagnosed and treated early retained a visual acuity of 0.4 and 0.5, respectively, whereas in the other two patients, whose diagnosis and therapy were delayed (> 6 weeks), visual acuity was reduced to light perception. We conclude that use of the PCR in the intraocular fluids together with detection of autochthonous antibodies in the vitreous seem to be the most important diagnostic laboratory tools.(ABSTRACT TRUNCATED AT 250 WORDS)

Aged↗
PubMed1991

Colonization of adrenal glands and ovaries of mice by variants of HSV 1 and 2. II. Histopathological, immunohistochemical and in situ hybridization studies.

The herpes simplex virus (HSV)-infected mouse model was used to correlate histopathological lesions in adrenal glands and ovaries with the localisation of viral nucleic acids and viral antigens, employing in situ hybridization and immunohistochemistry. In the adrenals, the lesions were mainly restricted to the zona fasciculata and the zona reticularis, sometimes extending to the medulla. In the ovaries, lesions were detected in follicles and in the stroma. During the course of infection, HSV nucleic acids could be detected earlier than HSV proteins. Next to the center of necrotic foci mainly HSV proteins were detected, whereas peripheral cells were found to contain viral nucleic acids. In situ hybridization revealed no proof of HSV latency in either organ. Among HSV-1 and HSV-2 strains of different neurovirulence, only HSV-2 variant ER- failed to replicate in adrenal glands and ovaries, whereas the neuroinvasive variant ER+ showed the same patterns as the HSV-1 strains used.

Adrenal Glands↗
PubMed1990

Epstein-Barr virus infection rapidly progressing to monoclonal lymphoproliferative disease in a child with selective immunodeficiency.

We report on a 30-month-old previously healthy Turkish boy who presented with fever, hepatosplenomegaly and generalized lymphadenopathy. He died 4 months after admission in spite of treatment with steroids, acycloguanosine and cyclophosphamide. Epstein-Barr virus (EBV) DNA was detected in the patient's bone marrow and in a lymph node biopsy. Cells from the lymph node biopsy showed monoclonal rearrangements of immunoglobulin heavy chain genes but no rearrangements of T-cell receptor beta-chain genes or immunoglobulin kappa chain genes. Serological data indicated chronic active EBV infection. There was a slight increase of CD8 positive cells in peripheral blood and a normal response to T-cell mitogens. However, T-cell lines established with interleukin 2 from lymph node biopsy completely failed to kill autologous EBV-transformed B-cells and K 562 target cells. Moreover, in regression tests the patient's peripheral blood mononuclear cells completely failed to limit outgrowth of autologous EBV infected B-cells. We conclude that the patient's selective immuno-deficiency had led to the rapid development of EBV-associated monoclonal lymphoproliferation.

Antibodies, Viral↗
PubMed1989

Structural analysis of proviral DNA in simian foamy virus (LK-3)-infected cells.

Proviral DNA of the T-cell lymphotropic simian foamy virus strain LK-3 was characterized. In infected cells, multiple copies of unintegrated linear duplex viral DNA of about 13 kbp length are present. Nuclease S1 treatment of the DNA generated two fragments of 6.5 and 6.0 kbp length that were cloned in phage and plasmid vectors. The proviral DNA contains a single-stranded gap of 109 nucleotides. A DNA fragment spanning the gap was cloned after completing the double strand by DNA synthesis in vitro. At the 3' end, the gap contains a polypurine tract (PPT) similar to the putative initiation site of retroviral plus strand DNA synthesis, suggesting discontinuous DNA synthesis. Further analysis of the genome architecture revealed LTRs of 1.7 kbp length. An additional 1.7 kbp DNA fragment was detected after nuclease S1 digestion of proviral DNA and probably represents trimmed intermediates of "strong-stop" DNA.

Animals↗
PubMed1988

Fatal Hodgkin and non-Hodgkin lymphoma associated with persistent Epstein-Barr virus in four brothers.

Three brothers from one family died of Hodgkin disease and a fourth brother from a diffuse malignant non-Hodgkin lymphoma. This patient exhibited a constant deficiency of serum immunoglobulins and elevated antibody titers to Epstein-Barr viral antigens. Epstein-Barr virus DNA sequences were detected in DNA isolated from lymph node biopsies from two of the patients. Initially, no abnormalities in the numbers of B and T cells could be detected. Peripheral blood lymphocytes of the patients did not react in the mixed lymphocyte culture assay. We suggest that an immune deficiency to Epstein-Barr virus may favor the proliferation of malignant lymphocytes after Epstein-Barr viral infection. Monoclonal lymphoid B cell lines established spontaneously in vitro from a lymph node biopsy specimen and from peripheral blood specimens from two of the patients. The cells harbor Epstein-Barr viral DNA sequences in multiple genome equivalents and express Epstein-Barr nuclear antigen. The cells contain a tenfold increased level of c-fgr-related RNA transcripts compared with peripheral blood lymphocytes of healthy adults. No obvious amplifications or translocations of the c-myc, c-abl, or c-fgr gene could be detected.

Adult↗
PubMed1988

Heterogeneity of primate foamy virus genomes. Brief report.

DNA of the T-lymphotropic simian foamy virus (SFV) LK-3 was cloned in a plasmid vector and used as a probe in comparative DNA:DNA hybridization studies with primate foamy viruses (SFV serotypes 1, 2, 3, 5, 6, 7, 8, fresh SFV isolates from 12 African green monkeys and one rhesus monkey, as well as human syncytium forming virus). All freshly isolated viruses seemed to be variants of SFV types 2 and 3. Especially LK-3 appeared to be closely related to SFV type 3. No hybridization of the LK-3 clone was observed to DNA sequences of human immunodeficiency virus (HIV-1).

Animals↗
PubMed1987

The potency of interferon-alpha 2 and interferon-gamma in a combination therapy of dendritic keratitis. A controlled clinical study.

Forty-five patients with virologically confirmed dendritic keratitis were treated in a randomized, double-blind controlled study with a basic therapy of trifluorothymidine (TFT) eye drops. In addition they received different human recombinant interferon (rHu IFN) eye drops. The following results were obtained for average healing times: TFT plus one drop daily of rHu IFN-alpha 2 arg (30 million iu/ml): 3.3 days, TFT plus rHu IFN-gamma (30 million iu/ml): 3.9 days, TFT plus a mixture of alpha plus gamma (0.3 million iu/ml each): 6.1 days, TFT plus a mixture of alpha plus gamma (1.5 million iu/ml each): 3.3 days. High-titer gamma interferon did not significantly differ from high-titer alpha interferon in the combination therapy of dendritic keratitis. A mixture of alpha plus gamma at a moderate titer (1.5 million iu/ml each) was as effective as a high-titer mono-preparation. Adding a low-titer interferon mixture gave no better therapeutic results than antiviral monotherapy. Thus it seems possible to save about 90% of interferon commonly used in the combination therapy of dendritic keratitis by applying a mixture of different suitable interferons instead of interferon monospecies.

Clinical Trials as Topic↗
PubMed1987

[Cytomegaloviruses and Kaposi sarcoma].

The cause of Kaposi's sarcoma is still unknown. Apart from genetic influences, the data available at present indicate that cytomegaloviruses may play a certain role. Serological investigations as well as results from molecular biology techniques seem to point to an association. However, whether the association is indeed causal remains to be determined.

Cell Line↗
PubMed1986

Nucleic acid hybridization for detection of herpes viruses in clinical specimens.

A diagnostic hybridization assay for detecting varicella zoster virus (VZV), herpes simplex virus (HSV), and Epstein-Barr virus (EBV) in different clinical specimens was developed using cloned viral DNAs as probes. All probes detected at least 5 pg of homologous DNA and did not cross-react with other viral or cellular DNA. Results of cell culture, serology, and DNA assay were highly concordant. Using a simple standardized protocol for preparation of specimens, hybridization, and washing procedures, this sensitive and specific assay appears to be useful for screening clinical specimens and may be helpful in confirming the serological diagnosis of HSV encephalitis and persistent EBV infections or EBV-associated diseases.

Adult↗
PubMed1986

Detection of herpes simplex virus and adenovirus DNA by dot blot hybridization using in vitro synthesized RNA transcripts.

A new diagnostic assay was developed to detect herpes simplex virus (HSV) and adenovirus DNA in clinical specimens using in vitro synthesized radioactively labelled RNA transcripts from virus-specific DNA fragments cloned in transcription vector pSP 64/65. RNA probes derived from HSV-I-Eco-RI-G-DNA fragment show a sensitivity of less than 3 pg of whole plasmid DNA and hybridize only with DNA of HSV I and II, but not with other viral or cellular DNA. The analysis of 15 clinical specimens showed concordance with virus isolation, except for two culture-negative samples of cerebrospinal fluid of patients with suspected HSV encephalitis, which was confirmed by serology as well as by hybridization. Using RNA transcripts from adenovirus-2-Hind-III-D-DNA fragment, we attained a sensitivity of less than 3 pg of whole plasmid DNA. This probe detected different types of adenovirus, but failed to hybridize to other viral, bacterial or cellular DNA. Compared with the cell culture method this assay did not show any false-positive or false-negative results in 16 different clinical specimens. The technique is sensitive, specific and useful for screening clinical specimens and may be helpful in confirming the diagnosis of HSV encephalitis.

Adenovirus Infections, Human↗
PubMed1986

Detection of human cytomegalovirus in urine by DNA-DNA and RNA-DNA hybridization.

A diagnostic hybridization assay for detecting human cytomegalovirus (HCMV) DNA in urine specimens was developed by using cloned viral DNA and in vitro-synthesized RNA probes. Both probes detected 3 pg of homologous DNA and hybridized with DNA of HCMV but not with other viral or human cellular DNA tested. In 95 urine specimens simultaneously tested by cell culture, the sensitivity of hybridization was at least 83%, and the specificity was at least 92%. This assay will be useful for rapid viral diagnosis with wide clinical applications such as screening of immunocompromised patients and quantitation of viral shedding in patients with primary or reactivated HCMV infection who may be receiving antiviral therapy.

Child↗
PubMed1986

Detection and characterization of infectious DNA intermediates of a primary foamy virus.

DNA from human T-lymphoid (Molt-4) and hamster kidney (BHK-21) cells infected with the T-lymphotropic simian foamy virus LK-3 was shown to be infectious, when assayed by transfection of BHK-21 cells. The proviral genome was further characterized by blot hybridization to a specific cDNA probe, which had been prepared by reverse transcription in vitro using viral RNA and RNA-dependent DNA polymerase present in cytoplasmic extracts of infected BHK-21 cells. This probe hybridized to a DNA species of 14 kbp in extracts from LK-3-infected diploid human fibroblasts, Molt-4 and BHK-21 cells, whereas no hybridization occurred with DNA from the respective uninfected controls. No integrated proviral DNA could be demonstrated, and the 14 kbp DNA was shown not to represent circular DNA. The patterns of restriction endonuclease and S1 nuclease fragments indicated a unique configuration of linear double-stranded DNA containing a single-stranded section separating two subunits one of which may be sufficient to transmit LK-3 by transfection with DNA.

Animals↗
PubMed1985

Interferon therapy for bronchial papillomatosis controlled by papillomavirus-DNA hybridization.

In biopsy specimens from a patient with tracheal and bronchial papillomatosis, human papillomavirus (HPV) type 11 DNA was identified. Treatment with leukocyte interferon was initiated, and the results of therapy was monitored by molecular hybridization of biopsy specimens with phosphorus 32-labeled HPV type 11 DNA after interferon application. There was no improvement of the clinical course, although two of five biopsy specimens showed a considerable reduction in the amount of HPV type 11. We discuss the reasons for the insufficient effect of therapy and propose molecular hybridization as an additional method for therapy control in viral diseases.

Bronchial Neoplasms↗