Biomedical subjects

E F Hunter

Publications and source records attributed to E F Hunter.

At least 19 recordsLinked to original sources

PubMed2000

Telephone support for persons with chronic mental illness.

These cases offer evidence of the potential utility of telephone support for one of the most challenging segments of the population. This group of clients often does not have strong support of any kind. Telephone support shows promise of offering cost-effective care for persons with psychiatric disabilities. Home healthcare nurses are encouraged to use this information as a basis for exploring the use of telephone support as a cost-effective system with their patients.

Community Health Nursing↗

PubMed1992

Specific immunofluorescent staining of pathogenic treponemes with a monoclonal antibody.

Two hybrid cell lines which produced mouse monoclonal antibody to the DAL-1 street strain of Treponema pallidum subsp. pallidum were established. These monoclonal antibodies strongly reacted with T. pallidum subsp. pallidum (Nichols strain, DAL-1, and two other street strains, strains MN-1 and MN-3) and T. pallidum subsp. pertenue by indirect microimmunofluorescent antibody and enzyme-linked immunosorbent assay techniques, but they did not react with normal rabbit testicular tissue. These monoclonal antibodies did not react with nonpathogenic treponemes, such as T. phagedenis Reiter, T. denticola MRB, T. refringens Noguchi, or other spirochetes, such as Borrelia burgdorferi and Leptospira interrogans serovar pomona in microimmunofluorescent antibody smear slides or in Western blots (immunoblots). While unlabeled antibodies are useful for investigating the antigenic structures of T. pallidum, we labeled these monoclonal antibodies with fluorescein isothiocyanate and used them for diagnosing syphilis by direct staining of lesion exudate or T. pallidum subsp. pallidum in formalin-fixed tissues from patients suspected of having syphilis. Both monoclonal antibodies were directed against antigens of T. pallidum subsp. pallidum with a molecular weight of 37,000 as determined by the Western blotting technique.

Animals↗

PubMed1992

Atriopeptin: an endogenous corticotropin-release inhibiting hormone.

Activation of the hypothalamic-pituitary-adrenocortical axis is a major component of the body's response to stress. Current theories on the pathophysiology of disorders associated with hyperfunction of the axis, such as depression and Cushing's stress, are based on the concept that anterior pituitary adrenocorticotropin (ACTH) secretion is stimulated by hypothalamic corticotropin-releasing hormones and inhibited by adrenal corticosteroids. Hypothalamic inhibitory control of pituitary ACTH secretion has been also postulated, but has not gained general acceptance because of the lack of definitive evidence for a corticotropin-release inhibiting hormone. It is shown here that in conscious rats stress-induced secretion of ACTH and corticosterone is markedly enhanced by the immunoneutralisation of atriopeptin. Therefore, we propose that atriopeptin is a physiologically relevant corticotropin-release inhibiting hormone.

Adrenal Cortex↗

PubMed1991

Specific immunofluorescence staining of Treponema pallidum in smears and tissues.

To date, tissue sections prepared from Formalin-fixed tissues have not been successfully stained with Treponema pallidum subspecies-specific antibody in a direct fluorescent-antibody assay. While current methods stain T. pallidum, they do not distinguish T. pallidum from other spirochetes such as Borrelia burgdorferi (E. F. Hunter, P. W. Greer, B. L. Swisher, A. R. Simons, C. E. Farshy, J. A. Crawford, and K. R. Sulzer, Arch. Pathol. Lab. Med. 108:878-880, 1984). Because trypsin pretreatment of tissue sections has enhanced other immunofluorescent-antibody (IFA) applications, we compared the use of the trypsin digestion method with the current 1% ammonium hydroxide (NH4OH) method as a means to obtain specific staining of T. pallidum in tissues by both direct and indirect IFA techniques. Pretreated T. pallidum-infected tissues sections from rabbits, hamsters, and humans were quantitatively examined with the direct fluorescent-antibody-T. pallidum test conjugate absorbed with Treponema phagedenis, the Reiter treponeme. For indirect staining, a serum specimen from a patients with syphilis absorbed by affinity chromatography with T. phagedenis was used as the primary reagent, and a fluorescein isothiocyanate-labeled rabbit anti-human globulin was used as the secondary reagent. Serum specificity was established first by examining antigen smears of T. pallidum subsp. pallidum, T. pallidum subsp. pertenue, B. burgdorferi, T. phagedenis, and Treponema denticola MRB and then by examining tissues infected with these pathogens plus those infected with four Leptospira serovars. When we stained tissue using the direct IFA method that is currently a standard method for the examination of chancre smears, we found it to be unsuitable for use with tissue. Trypsin digestion did not offer an improvement over the NH4OH pretreatment method in the specific identification of T. pallidum by direct IFA. However, specific identification of T. pallidum in tissue sections was obtained by the indirect IFA technique after either trypsin or NH4OH pretreatment.

Animals↗

PubMed1990

Promoting productivity of individuals with psychiatric disorders in the work setting.

Just as physical supports have been provided for the physically disabled in the work environment, psychological supports are needed for the psychiatrically disabled. The occupational health nurse is the best trained and most cost effective health team member to develop the training programs that would provide these psychological supports in the work setting. A four phase comprehensive program is needed to destigmatize psychiatric disabilities and develop adequate support systems. Among the most prevalent psychiatric disabilities, episodes of symptom exacerbation can be managed with the help of a trained peer support person.

Costs and Cost Analysis↗

PubMed1986

Detection of immunoglobulin M in cerebrospinal fluid from syphilis patients by enzyme-linked immunosorbent assay.

Cerebrospinal fluid (CSF) samples were evaluated in an immunoglobulin M enzyme-linked immunosorbent assay (IgM ELISA) for syphilis with sonic extracts of Treponema pallidum coated on polystyrene plates. The ELISA procedure was reproducible, and T. pallidum antigens were stable., A total of 15 CSF samples from patients with neurosyphilis, 18 CSF samples from patients with syphilis, 12 CSF samples from patients treated for syphilis, and 494 CSF samples from patients with neurologic or other systemic diseases were tested. The IgM ELISA gave reactive results in all of six symptomatic and congenital neurosyphilitic patients and none of nine asymptomatic neurosyphilitic patients. Of 524 CSF samples from nonneurosyphilitic individuals, 513 were nonreactive, resulting in 98% test specificity. The IgM ELISA in CSF should prove to be useful for confirmation of symptomatic neurosyphilis.

Albumins↗

PubMed1985

Four-step enzyme-linked immunosorbent assay for detection of Treponema pallidum antibody.

Further studies of a four-step enzyme-linked immunosorbent assay procedure to detect Treponema pallidum antibody are described. High-titered antibody, produced in rabbits by intravenous injection of T. pallidum, was used to coat polyvinyl chloride microtiter plates. To these plates a known concentration of T. pallidum was added, followed in successive steps by serial dilutions of human sera and appropriately diluted peroxidase-labeled anti-human immunoglobulin G antibody. O-Phenylenediamine was the substrate. A total of 340 sera were obtained from the DeKalb County Sexually Transmitted Diseases Clinic, Atlanta, Ga., and examined within 3 days of receipt. Ninety-six percent test agreement between the enzyme-linked immunosorbent assay and the fluorescent treponemal antibody absorption-double staining test was obtained. A total of 372 additional sera stored at -20 degrees C were examined. The overall sensitivity of the enzyme-linked immunosorbent assay with sera from patients with various stages of syphilis was 96%. With sera from uninfected individuals, the specificity of the enzyme-linked immunosorbent assay was 95%. No antigen instability was noted with the two antigen preparations used during this evaluation.

Antibodies, Bacterial↗

PubMed1985

Rheumatoid factor in syphilis.

Immunoglobulin M (IgM) antibodies directed against IgG antibodies (rheumatoid factor [RF]) are known to occur often in patients with syphilis and to interfere with serological tests measuring specific antibodies of the IgM class. In this study we examined the occurrence and specificity of the RF and demonstrated a simple method to detect and eliminate the RF for a specific Treponema pallidum IgM enzyme-linked immunosorbent assay. We measured the occurrence of the RF with a sensitive enzyme-linked immunosorbent assay and found that it increased with the duration of syphilitic disease: 1 of 13 primary syphilis serum specimens, 3 of 13 secondary syphilis serum specimens, and 10 of 27 latent syphilis serum specimens were reactive in this RF test. Those sera containing IgM RF were immunoprecipitated with anti-human gamma chain antibodies and 2% polyethylene glycol until the RF was removed. One serum specimen from a patient in the secondary stage of syphilis and eight serum specimens from patients with latent disease still presented the RF after immunoprecipitation. Removal of the IgG antibodies also improved the sensitivity of the treponemal IgM test, indicating competition of these antibodies for binding sites of the antigen. The enzyme-linked immunosorbent assays for detection of RF and antitreponemal IgM antibodies are performed on the same plate. Theoretically, only sera positive for both tests have to be immunoprecipitated. But our findings indicated an increase in sensitivity of the IgM enzyme-linked immunosorbent assay after removal of IgG antibodies responsible for competition at the binding sites.

Antibodies, Anti-Idiotypic↗

PubMed1984

Double-conjugate enzyme-linked immunosorbent assay for immunoglobulins G and M against Treponema pallidum.

An enzyme-linked immunosorbent assay (ELISA) for the simultaneous measurement of immunoglobulin G (IgG) and IgM was developed to detect antibodies to Treponema pallidum. Wells of polystyrene microtiter plates were coated with T. pallidum antigen, diluted patient serum was added, and IgG and IgM which bound to the T. pallidum antigen were measured by the simultaneous addition of alkaline phosphatase-labeled anti-human IgG and horseradish peroxidase-labeled anti-human IgM. Bound IgG was detected first, followed by bound IgM. After development of the procedure, 145 categorized sera were evaluated: 60 from individuals without syphilis; 62 from patients with syphilis, including 22 with primary, 20 with secondary, and 20 with latent phases of syphilis; and 23 from patients with rheumatoid arthritis. Of the 60 sera from individuals without syphilis, 100% were nonreactive for IgG antibody and 16% were reactive for IgM. Of the 23 sera from patients with rheumatoid arthritis, 3 were reactive for IgG and 3 were nonreactive for IgM. Of the 62 sera from patients with syphilis, 61 (98%) were reactive for IgG antibody with increased titers as the stage of syphilis increased, whereas IgM reactivity decreased. This enzyme-linked immunosorbent assay appears to be a simple method for the simultaneous measurement of antibodies under equal assay conditions.

Antibodies, Bacterial↗

PubMed1984

Immunofluorescent staining of Treponema in tissues fixed with formalin.

Immunofluorescent examination of formalin-fixed tissue for Treponema pallidum has generally been unsatisfactory because of nonspecific background fluorescence and poor contrast. We examined the process of treating deparaffinized formalin-fixed tissue sections with 1% ammonium hydroxide (NH4OH) to improve fluorescent staining. Treponema pallidum- and Treponema pertenue-infected rabbit testes or human tissue biopsy specimens fixed in 10% buffered formalin and embedded in paraffin were examined. Sections were cut one week to five years after embedment. Tissues were then stained with fluorescein- or rhodamine-labeled human anti- T pallidum globulin for 30 minutes at 37 degrees C. Treponemes were consistently stained and background staining was generally reduced after NH4OH treatment in both fresh and stored tissue. Cutting sections at a thickness of approximately 2 micron was critical to achieve optimal fluorescence.

Ammonium Hydroxide↗

PubMed1983

Fluorescent treponemal antibody absorption double-staining test evaluation.

The fluorescent treponemal antibody absorption (FTA-ABS) double-staining (DS) test has been developed for microscopes equipped with incident illumination, and the procedure offers many advantages over the FTA-ABS test when tests are performed with this equipment. In this study, 346 fresh sera, including 35 from patients with syphilis, were evaluated by the FTA-ABS DS test. Parameters for investigation included two readers, each using a different microscope; a new FTA-ABS DS test reporting system; sera heated at 56 degrees C for 30 min versus unheated sera; and sera retested after at least 2 weeks of freezer storage. Agreement for FTA-ABS DS test readings between the two microscopes was 99%. Between-test agreement for the FTA-ABS test with the conventional reporting system and the FTA-ABS DS test with the new reporting system was 95%. Sensitivity calculations based on reactivity for the 35 syphilis sera were 94% for the FTA-ABS DS test and 91% for the FTA-ABS test. Specificity calculations based on non-reactivity of nonsyphilis sera were 98% for the FTA-ABS DS test and 93% for the FTA-ABS test. Differences in percentages appeared to be related to borderline readings in the FTA-ABS test. For example, if the same reporting system was used for the reference FTA-ABS test, the specificity was 97%. When sera were examined within 48 h, no difference was observed in results obtained with heated and unheated sera. Sera frozen for 2 weeks showed comparable results in the FTA-ABS DS test and the FTA-ABS test. These findings strongly support the recommendation that the FTA-ABS DS test be accepted as a confirmatory test for syphilis. The new reporting system for the FTA-ABS DS test would be advantageous for the reference FTA-ABS procedure.

Evaluation Studies as Topic↗

PubMed1982

Evaluation of the microenzyme-linked immunosorbent assay with Treponema pallidum antigen.

Whole-cell sonicates of Treponema pallidum, Nichols strain, were evaluated in an enzyme-linked immunosorbent assay (Elisa) for syphilis, and results were read in a Dynatek Microelisa Reader. The antigen was evaluated with sera from patients with syphilis, persons presumed normal, and biological false-positives. Two hundred and ninety-seven sera were tested by the ELISA with T. pallidum antigens, the Venereal Disease Research Laboratory (VDRL) slide test, the fluorescent treponemal antibody absorption (FTA-Abs) test, and the microhemagglutination assay for T. pallidum antibodies (MHA-TP). The results of all of the tests were compared. The ELISA, with 89.3% sensitivity, was less sensitive than the VDRL (93.3%) and FTA-Abs (100.0%) tests but more sensitive than the MHA-TP (76.0%). THe ELISA was considerably more sensitive in primary syphilis than the MHA-TP. Specificity was as follows: ELISA, 98.5%; FTA-Abs test, 97.8%; MHA-TP, 98.2%; and VDRL test, 92.7%. The ELISA has good potential as a confirmatory test in the serodiagnosis of syphilis.

Antibodies, Bacterial↗

PubMed1982

Sodium desoxycholate-extracted treponemal antigen in an enzyme-linked immunosorbent assay for syphilis.

The extraction of Treponema pallidum antigen with sodium desoxycholate, based on a previously described procedure (J. Portnoy and H.J. Magnuson, J. Immuno. 75:348-355, 1955), was used in an enzyme-linked immunosorbent assay (ELISA) test for syphilis. The antigen was prepared from T. pallidum street strain no. 14, and its overall sensitivity and specificity was compared with those of sonicated antigen preparations made with phosphate-buffered saline. The optimum serum dilution for testing and the significant absorbance reading at 490 nm were selected by examination of quantitative dilutions of 91 sera from presumably normal individuals and 92 sera from syphilitics. The time and temperature of serum and conjugate incubations were also examined. With an absorbance reading of greater than or equal to 0.2 at the 1:80 serum dilution, 88 (95.8%) of 92 sera from syphilitics were reactive in the ELISA test with desoxycholate-extracted antigen, and 82 (89.1%) were reactive with the sonicated antigen. Only one nonsyphilitic serum was reactive with each antigen. Greater sensitivity without loss in specificity was obtained with longer serum and conjugate incubations. We concluded that an ELISA test with sodium desoxycholate-extracted antigen is more sensitive than and equally specific to an ELISA with sonicated treponemal antigen.

Antigens, Bacterial↗

PubMed1982

Immunofluorescence and Treponema infection: a method using immunofluorescence to study rabbit testicular tissue infected with T pallidum and T pertenue.

We compared immunofluorescent staining of rabbit testicular tissue infected with Treponema pallidum or T pertenue, and fixed in Bouin's fixative, 95% cold ethyl alcohol with 1% glacial acetic acid, or routine 10% buffered formalin solution. The fixative of choice clearly was Bouin's. Although we studied only rabbit tissue, we assume that these fixatives will work well in human biopsy or autopsy material when identification of pathogenic Treponema is needed.

Animals↗

PubMed1981

Fluorescent treponemal antibody-absorption double-staining procedure.

The fluorescent treponemal antibody-absorption double-staining step-by-step procedure and proposed reference reagents for the test are described. The test and reagents were evaluated in two separate laboratories on 265 fresh sera, and test results were compared with the reference fluorescent treponemal antibody-absorption test results performed in a third laboratory. The data indicate that the tests are comparable in the areas where the test is recommended for use. Problems with inadequate light filtration occurred, but these could be resolved. This test is recommended for use with microscopes equipped with incident illumination.

Antibodies, Anti-Idiotypic↗

PubMed1980

Further studies with the fluorescent treponemal antibody-absorption double-staining procedure.

A double-staining procedure for the fluorescent treponemal antibody-absorption test, using fluorescein isothiocyanate as a label for the class-specific anti-human globulin and tetramethylrhodamine isothiocyanate as a label for a counterstain reagent, has been described. This method requires the addition of a KP560 barrier filter, with a microscope equipped with vertical illumination, to exclude the rhodamine emission in reading the fluorescein fluorescence. The present study evaluated reversing the dye label for each conjugate in the double-staining procedure, thus eliminating the need for the KP560 filter. It also considered the possibility of shortening the counterstaining time and compared various methods for preparing antigen slides in an attempt to establish a method that increases the number of treponemes per microscopic field. The results indicate that a rhodamine-labeled class-specific anti-human globulin as a primary stain, and a fluorescein-labeled anti-treponemal globulin as a counterstain, provide an acceptable method for performing the fluorescent treponemal antibody-absorption double-staining procedure. Nonfixed antigen slides were held for 16 days in a desiccator or stored in plastic bags with silica gel for 3 weeks; then, with methanol fixation, they were used satisfactorily in the double-staining procedure. A shortened incubation time for the counterstain allowed more rapid slide processing.

Antigens, Bacterial↗

PubMed1979

Problems affecting performance of the fluorescent treponemal antibody-absorption test for syphilis.

Immunofluorescent staining of Treponema pallidum was studied to clarify the effect of three factors on the results of the fluorescent treponemal antibody-absorption test: (i) heat inactivation of sera at 56 degrees C for 30 min before testing, (ii) use of multicircle slides, and (iii) tungsten illumination to visualize and assess unstained treponemes on reactive as well as nonreactive smears. It was found that serum inactivation before testing was not necessary for detection of immunoglobin G antibody, but an immunoglobulin M prozone was detected in unheated serum. On multicircle slides, it was demonstrated that a false-positive reaction could be obtained in 30 s at 37 and 25 degrees C if a smear where a nonreactive serum had been placed was crossed by a strongly reactive serum from another circle. Tungsten illumination proved necessary for correct assessment of unstained treponemes on all fluorescent treponemal antibody-aborption test smears, reactive or nonreactive. The possible role of these factors in incorrect fluorescent treponemal antibody-absorption test results is discussed.

Antibodies, Bacterial↗

PubMed1979

Double-staining procedure for the fluorescent treponemal antibody absorption (FTA-ABS) test.

The fluorescent treponemal antibody absorption (FTA-ABS) double-staining procedure was reproducible, comparable to the conventional test, and easy to read. We recommend the use of the FTA-ABS double-staining procedure for microscopes with incident illumination, the 100 x/1.30 oil achromatic objective and the 6.3 x ocular to obtain optimal fluorescence, and the KP560 as a barrier filter to exclude rhodamine emission when fluorescein fluorescence is read. With this system, errors related to poor focusing or failure to visualise treponemes on all smears should be eliminated.

Fluoresceins↗