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D Petzoldt

Publications and source records attributed to D Petzoldt.

At least 37 records · Page 2Linked to original sources

Measurement of free human leukocyte elastase and human leukocyte elastase/alpha 1 proteinase inhibitor complexes by an enzyme-linked immunosorbent assay.

We report on an ELISA procedure for the quantitative analysis of total human leukocyte (PMN)-elastase, i.e., the simultaneous determination of (i) free (non-complexed) PMN-elastase and of (ii) PMN-elastase that is complexed to alpha 1 proteinase inhibitor. Simultaneous detection of both forms of PMN-elastase was achieved using a monoclonal antibody that recognizes a PMN-elastase-specific epitope present on both the uncomplexed and the complexed form of the enzyme. The test system described is reliable, easy to perform and permits the determination of total PMN-elastase in complex biological fluids such as plasma or seminal fluid. It is to be expected that this test system will be useful for investigations of human PMN-elastase in biological specimens obtained from both normal and pathological conditions.

Antibodies, Monoclonal

Current status of serotyping of Neisseria gonorrhoeae.

Protein I has become the basis of serotyping of N. gonorrhoeae. A panel of 12 anti-protein I monoclonal antibodies is supplied by Syva Company, Palo Alto, CA, through distributors for research purposes only. These distributors are responsible for storing the monoclonal antibodies, supplying research groups and training new users, particularly in the interpretation of coagglutination reactions. A European Workshop was held in Heidelberg, Federal Republic of Germany in July 1988 to discuss serotyping with these antibodies. The current uses of serotyping of N. gonorrhoeae include epidemiological studies, clinical purposes and surveillance of antibiotic resistance and plasmid carriage. Predominant serovars may be subtyped either by additional antibodies or with the use of another technique, such as auxotyping, determination of antibiotic sensitivities, plasmid analysis or genetic fingerprinting. Conversely, there is growing evidence that it may be appropriate to group certain serovars together. Data collected during prevalence studies could be combined with clinical information and an international data bank set up. This supposes a close future collaboration of all groups involved in the epidemiology of N. gonorrhoeae.

Antibodies, Monoclonal

Characterization of Borrelia burgdorferi associated antigens by monoclonal antibodies.

In this paper, we present a series of murine mAb recognizing B. burgdorferi antigens. The antibodies were characterized by immuno-blotting and immuno-fluorescence studies using isolates of B. burgdorferi from North America and Europe, respectively. Moreover, reactivity of the antibodies with recombinant B. burgdorferi flagellin and OspA was studied. The results suggest these anti-B. burgdorferi mAb as valuable tools for the serological analysis of B. burgdorferi isolates and for affinity-purification of the respective proteins. Moreover, these mAb appear suitable to classify antigenic variants of B. burgdorferi and to study the protective capacity of antibodies in a murine model for B. burgdorferi infection.

Animals

[Nucleic acid hybridization for direct detection of Chlamydia trachomatis. Comparison of a radioactive with a non-radioactive procedure].

A total of 234 urogenital specimens were investigated with a radioactive and a nonradioactive rRNA:cDNA-hybridization test for the direct detection of Chlamydia trachomatis (Gen-Probe, San Diego, Calif.). The results were compared with those yielded by the conventional cell culture technique (McCoy cells). Using cell culture a urogenital C. trachomatis infection could be detected in 37 of 234 patients (15.8%) at two outpatient sexually transmitted disease clinics. Referred to the detection rate, cell culture sensitivity was 83.8% for the radioactive test and 86.5% for the nonradioactive DNA-hybridization test. The specificity of the radioactive test was 94.8% and that of the nonradioactive test, 93.4%.

Cells, Cultured

[High-level tetracycline-resistance of Neisseria gonorrhoeae].

High level tetracycline resistance of Neisseria gonorrhoeae is mediated by a 25.2-MD plasmid and is characterized by a minimal inhibitory concentration of 16 micrograms/ml or more. The 25.2-MD plasmid originated through the insertion of the streptococcal tetM determinant into the 24.5-MD transfer plasmid of N. gonorrhoeae. In the present study the prevalence of N. gonorrhoeae with high-level tetracycline resistance in the Heidelberg gonococcal population between 1981 and 1989 was determined. Among 1765 N. gonorrhoeae isolates, one strain with high-level tetracycline resistance was identified. The presence of the 25.2-MD plasmid in this strain was demonstrated by plasmid-agarose gel electrophoresis. The auxotype/serovar class proline-/IB-1 indicates the importation of this strain from the USA. The 25.2-MD recombinant plasmid, in contrast to the 24.5 MD transfer plasmid, has a rather wide host range, which is favourable to the occurrence both of high-level tetracycline resistance and of beta-lactamase plasmids in N. meningitidis.

Bacterial Proteins

[Tumoral calcinosis].

Tumoral calcinosis is a distinct entity, which is rarely seen in Europe and North America but much more common in black Africans. Typical symptoms are calcified nodules, which grow while remaining asymptomatic and are found in the tissues adjacent to the large joints of the body. Histologically there is collagen necrobiosis initially, which results in aggregates of densely calcified material. The aetiology is unknown, but the condition is probably a form of dystrophic calcification caused by mechanical injury. A patient with tumoral calcinosis is presented, and the clinical and histological findings are described.

Aged

Netherton's syndrome: ultrastructure of the active lesion under retinoid therapy.

A young female patient, expressing the symptom triad of Netherton's syndrome, i.e., ichthyosis linearis circumflexa Comèl, trichorrhexis invaginata and other hair shaft defects, and atopic diathesis, has been treated successfully with the new retinoid preparation Etretin. Our electron microscopical study especially focused on the ultrastructural effect on the characteristic, active part of the skin lesions, which is only found within a narrow borderline just preceding the lesion's margin. In untreated skin, this part is characterized by dermal inflammation, immigrating inflammatory cells, and specific keratinization disturbances: synthesis of keratinization proteins is suppressed, serum exudates invade the epidermis, either filling the intercellular spaces of the upper spinous and the granular layer as finely granular, amorphous material, or they are partly phagocytosed and lie within intracellular, round-oval inclusions. The portions of the lesions lying towards the center are unspecific and represent recovery stages, ultrastructurally resembling stages of normal wound repair. Oral therapy with Etretin did not heal the basic defect, but drastically reduced exoserosis and the deposition of intra- and extracellular material. Keratinization seemed to normalize. The condition of the hair was also improved.

Acitretin

The significance of the ipazyme IgA and IgG antibody test in the diagnosis of urogenital chlamydial infections.

Urogenital specimens from 200 male and female patients were cultivated for the detection of C. trachomatis. For comparison, serum of the same patients was investigated with the Ipazyme IgA and IgG test. This comparison of culture and serological tests revealed a sensitivity of the IgA Ipazyme test of 56% and a specificity of 81%. For the IgG Ipazyme test, the corresponding values were 77% and 39%. The sensitivity of the conventional immunofluorescence test reached 46% and its specificity was 50% for the same group of patients. Antibiotic treatment of 13 IgA-positive patients resulted in a significant decrease of the titre in only one case. The introduction of the Ipazyme test does not open a new aspect in chlamydial serology, i.e. the diagnostic value of serology for the detection of a current chlamydial infection remains low.

Antibodies, Bacterial

Evaluation of a non-radioactive DNA probe for confirmatory identification of Neisseria gonorrhoeae.

A biotinylated DNA probe combined with a streptavidin-peroxidase complex for the identification of culture isolates of N. gonorrhoeae (Ortho diagnostic systems, Neckargemünd, FRG) was compared with the conventional carbohydrate utilisation test as reference. All 118 strains identified by the reference method as N. gonorrhoeae also gave positive reactions with the DNA hybridisation assay. However, with this test 2 of 23 non-gonococcal Neisseria or Branhamella species were identified as N. gonorrhoeae as well. The study shows that the DNA hybridisation technique can principally be used for the confirmatory identification of N. gonorrhoeae, but since specificity is particularly essential for confirmatory identification, the DNA hybridisation assay evaluated cannot be recommended for routine diagnosis.

Carbohydrate Metabolism

[Chlamydia trachomatis, a pathogen in infections of the upper respiratory tract?].

In neonates and in older children C. trachomatis can cause otitis media in rare cases. This fact has to be taken into account in diagnosis. The detection of C. trachomatis in the pharynx after orogenital sexual practice seems to be rather due to colonization than to an infection. According to existing studies C. trachomatis does not play a causative role in the pathogenesis of pharyngitis in adults.

Adult

Evaluation of a radioactive rRNA:cDNA-hybridisation assay for the direct detection of Chlamydia trachomatis in urogenital specimens.

A radioactive cDNA probe complementary to chlamydial ribosomal RNA was used to detect C trachomatis in urogenital specimens. Of 37 specimens positive with cell culture 31 were confirmed by the rRNA:cDNA hybridisation test, the sensitivity being 83.8%. The specificity of the hybridisation test was 94.4%, as 186 of 197 specimens that were negative by cell culture were also negative when assessed by the hybridisation method. Given a prevalence of 15.8% the predictive values for positive and negative results were 73.8% and 96.9%, respectively. In additional experiments the possible role of microorganisms added to the specimen collection medium was investigated. However, no indication for crosshybridisation was found; at high concentrations microorganisms interfered with the test procedure.

Cervix Uteri

Low concentrations of acid-soluble thiol (cysteine) in the blood plasma of HIV-1-infected patients.

Blood plasma samples from HIV-1-infected persons contain elevated glutamate concentrations up to 6-fold the normal level and relatively low concentrations of acid-soluble thiol (i.e. decreased cysteine concentrations). The intracellular glutathione concentration in peripheral blood-mononuclear cells (PBMC) and monocytes from HIV antibody-positive persons are also significantly decreased. Therapy with azidothymidine (AZT) causes a substantial recovery of the plasma thiol levels; but glutamate levels remain significantly elevated and intracellular glutathione levels remain low. Cell culture experiments with approximately physiological amino-acid concentrations revealed that variations of the extracellular cysteine concentration have a strong influence on the intracellular glutathione level and the rate of DNA synthesis [( 3H]thymidine incorporation) in T cell clones and human and murine lymphocyte preparations even in the presence of several-fold higher cystine and methionine concentrations. Cysteine cannot be replaced by a corresponding increase of the extracellular cystine or methionine concentration. These experiments suggest strongly that the low cysteine concentration in the plasma of HIV-infected persons may play a role in the pathogenetic mechanism of the acquired immunodeficiency syndrome.

Acquired Immunodeficiency Syndrome

[Successful retinoid therapy of Netherton syndrome].

A young patient with Netherton's syndrome characterized by the classic triad of ichthyosis linearis circumflexa, trichorhexis invaginata and atopy was treated with Acitretin, a new retinoid preparation: 35 mg Acitretin/day resulted in a severe, erosive dermatitis which necessitated interruption of therapy. Even with 10 mg/day the patient had intolerable irritation of the integument. After a further dosage reduction to 5 mg/day there were no obvious side effects and a long-term treatment was possible, resulting in an obvious reduction of the ichthyotic lesions and improved hair growth. Electron microscopy in the active part of the skin lesions from untreated skin revealed granular, membrane-enclosed material intracellularly and in the intercellular spaces of the granular layer. Keratinization was almost completely suppressed. Therapy with Acitretin drastically reduced the deposition of intra- and extracellular material and normalized keratinization. Our results underline the importance of starting retinoid therapy in Netherton's syndrome at a low dosage and adjusting it carefully in each case with reference to the skin manifestations and the side effects.

Acitretin

Effects of antimicrobial therapy on sperm-mucus interaction.

Sperm-mucus interaction under in-vitro or in-vivo conditions might be affected by microorganisms colonizing the reproductive tract. In order to study the influence of antimicrobial therapy, an extensive microbial screening was performed including Chlamydia trachomatis, Mycoplasma hominis, Ureaplasma urealyticum, Neisseria gonorrhoeae, a broad spectrum of potentially pathogenic aerobic and anaerobic bacteria, Trichomonas vaginalis, herpes simplex virus and yeasts. One-hundred-and-six couples with a mean duration of infertility of 5.5 years (range 1-12 years) and with isolation of potentially pathogenic microorganisms in semen samples and/or cervical swabs were submitted to a prospective pilot study. None of the patients displayed signs or symptoms of infection in the lower genital tract. Before and after specific therapy, based on antimicrobial susceptibility testing, sperm analyses and in-vitro sperm penetration meter tests (SPMT) (Kremer) were performed. SPMT was evaluated with cervical mucus of patients' wives, collected after a standardized oral treatment with oestrogens and, additionally, in a crossed manner with cervical mucus and spermatozoa of fertile donors. The success of antimicrobial therapy was controlled by repeating the same microbial screening and was 96%. However, there was a marked change in the microbial pattern. A comparison of the results of sperm analyses before and after treatment revealed neither significant differences for sperm volume, sperm count, propulsive motility, morphology, vitality, pH, fructose concentration or number of round cells, nor was there a significant influence on the cervical index and the number of leukocytes in cervical mucus.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult

Evaluation of non-radioactive in situ hybridisation method to detect Chlamydia trachomatis in cell culture.

A DNA probe combined with a non-radioactive stain was used to detect inclusions of Chlamydia trachomatis in cell culture. Of 39 positive cultures detected by monoclonal antibodies in combination with direct immunofluorescence, 35 were positive by the DNA hybridisation method, the sensitivity being 89.7%. Staining with iodine showed a sensitivity of 87.2%, corresponding to 34 positive cultures. The specificity of DNA hybridisation method was 100%, as all 162 cultures that were negative by the immunofluorescence method were also negative when assessed by the DNA hybridisation method.

Cells, Cultured