scholarly journals ULTRASTRUCTURAL IMMUNOCYTOCHEMISTRY WITH UNLABELED ANTIBODIES AND THE PEROXIDASE-ANTIPEROXIDASE COMPLEX A TECHNIQUE MORE SENSITIVE THAN RADIOIMMUNOASSAY

1973 ◽  
Vol 21 (9) ◽  
pp. 825-833 ◽  
Author(s):  
GWEN C. MORIARTY ◽  
C. MICHAEL MORIARTY ◽  
LUDWIG A. STERNBERGER
Keyword(s):  
Staining Intensity ◽  
Poor Quality ◽  
Immunocytochemical Staining ◽  
Great Promise ◽  
Electron Microscopic ◽  
Immunocytochemical Technique ◽  
Titration Curves ◽  
Rat Pituitary ◽  
Normal Rat ◽  
Unlabeled Antibody

Titration curves were developed with antisera to 17-39ACTH (adrenocorticotropin) and 1-39ACTH with the techniques of radioimmunoassay and electron microscopic immunocytochemistry. For the latter method, the unlabeled antibody peroxidase-antiperoxidase complex immunocytochemical technique was used to stain normal rat pituitary intermediate lobes. By radioimmunoassay standards, the 17-39ACTH antiserum was of poor quality. Its titration curve exhibited a flat slope and it did not bind a significant amount of labeled antigen beyond a 1: 30 dilution. However, immunocytochemical staining was detected with this antiserum at dilutions as high as 1:1,500. The antiserum to 1-39ACTH was of better quality by radioimmunoassay standards. It bound 45% of the labeled antigen at a dilution of 1:5,000. Immunocytochemical staining intensity was nearly maximal at a 1:5,000 dilution and decreased progressively to a limiting value at 1:16,000. However, when incubation times in the antisera were increased from 3 min to match those of the radioimmunoassay (48 hr) maximal staining was achieved at dilutions as great as 1:512,000 where only trace amounts of the labeled antigen were bound in the radioimmunoassay. It was concluded that the unlabeled antibody peroxidase-antiperoxidase complex immunocytochemical technique was sensitive enough to detect antibodies in sera of low titer and/or avidity which are not detected by a radioimmunoassay. The technique holds great promise for a sensitive assay system.

scholarly journals ELECTRON MICROSCOPIC STUDY OF THE ADRENOCORTICOTROPIN-PRODUCING CELL WITH THE USE OF UNLABELED ANTIBODY AND THE SOLUBLE PEROXIDASE-ANTIPEROXIDASE COMPLEX

1972 ◽  
Vol 20 (8) ◽  
pp. 590-603 ◽  
Author(s):  
G. C. MORIARTY ◽  
N. S. HALMI
Keyword(s):  
Secretory Granules ◽  
Staining Intensity ◽  
Electron Microscopic ◽  
Early Effect ◽  
Acth Cell ◽  
Maximal Diameter ◽  
Soluble Peroxidase ◽  
Ultrathin Sections ◽  
Acth Secreting ◽  
Unlabeled Antibody

The technique involving use of unlabeled antibody and the peroxidase-antiperoxidase complex was used to identify the adrenocorticotropin (ACTH)-secreting cell in the anterior pituitary lobe of the rat and to localize ACTH in it electron microscopically in ultrathin sections. The ACTH cell is star-shaped, with processes extending around other cells, and contains secretory granules of a maximal diameter of 300 mµ arranged peripherally along the plasma membrane. Stain was observed on secretory granules, around them, in the Golgi complex and in rough endoplasmic reticulum. One day after adrenalectomy, the ACTH cell is degranulated and the staining intensity of its remaining granules and cytoplasm is decreased, suggesting release of ACTH stores. If cortisol is given 6 hr after adrenalectomy, 18 hr later the ACTH cells are well granulated and the granules stain more intensely than normal. In addition, staining around the granules and throughout the cytoplasm is more intense, suggesting that an early effect of cortisol is to block release of ACTH. Twenty-one days after adrenalectomy, the ACTH cells are greatly increased in numbers and have complex, tortuous processes filled with intensely stained secretory granules.

scholarly journals THE UNLABELED ANTIBODY ENZYME METHOD OF IMMUNOCYTOCHEMISTRY QUANTITATIVE COMPARISON OF SENSITIVITIES WITH AND WITHOUT PEROXIDASE-ANTIPEROXIDASE COMPLEX

1974 ◽  
Vol 22 (8) ◽  
pp. 782-801 ◽  
Author(s):  
JOHN P. PETRALI ◽  
DENNIS M. HINTON ◽  
GWEN C. MORIARTY ◽  
LUDWIG A. STERNBERGER
Keyword(s):  
Secretory Granules ◽  
Enzyme Reaction ◽  
Fold Increase ◽  
Immunocytochemical Staining ◽  
Intermediate Lobe ◽  
Reaction Products ◽  
Electron Microscopic ◽  
Specific Staining ◽  
Enzyme Method ◽  
Unlabeled Antibody

Araldite sections of rat pituitary intermediate lobe were used with anti-17-39 adrenocorticotropin in the unlabeled antibody enzyme method to compare electron microscopic immunocytochemical staining by peroxidase-antiperoxidase complex (PAP) with antiserum or purified antibody to peroxidase followed by peroxidase (PO). Quantitation of normalized optical densities of secretory granules offered high significance comparison (P < 0.0001) of experimental with control values and of experimental values with each other. Use of purified antibody (prepared by a new density gradient method) yielded four times higher sensitivity than antiserum to PO, while a 6.5-fold increase would have been expected from the degree of contamination of anti-PO in the serum by nonanti-PO immunoglobulin. Use of PAP was four to five times more sensitive than purified anti-PO and 20 times more sensitive than antiserum to PO. The increased sensitivity of PAP is explained by the high over-all binding affinity of PO for anti-PO in the cyclic PAP molecule, thus preventing the losses of PO that occur during washing when anti-PO and PO have been applied in sequence. Identification of the characteristic, cyclic PAP molecules affords confirmation of specific staining at high resolution. In the sequential application of anti-PO and PO, no PAP molecules are formed, thus making distinction of specific from nonspecific deposition of enzyme reaction products ambiguous. With the use of anti-17-39 ACTH and the intermediate lobe, the unlabeled antibody enzyme method was 16,000-100,000 times more sensitive than radioimmunoassay.

scholarly journals Four unlabeled antibody bridge techniques: a comparison.

1981 ◽  
Vol 29 (12) ◽  
pp. 1397-1404 ◽  
Author(s):  
P Ordronneau ◽  
P B Lindström ◽  
P Petrusz
Keyword(s):  
Horseradish Peroxidase ◽  
Gamma Globulin ◽  
Staining Intensity ◽  
Reaction Products ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
Immunocytochemical Techniques ◽  
Unlabeled Antibody ◽  
Shape And Size

Four unlabeled antibody immunocytochemical techniques, the "single bridge" (Avrameas S: Immunocytochemistry 6:825, 1969; Mason TE, Phifer RF, Spicer SS, Swallow RS, Dreskin RD: J Histochem Cytochem 17:190, 1969a; Sternberger LA, Cuculis JJ: 1969), the "single peroxidase-antiperoxidase (PAP)" (Sternberger LA, Hardy PH Jr, Cuculis JJ, Meyer HG: J Histochem Cytochem 18:315, 1970), the "double PAP" (Vacca LL, Rosario SL, Zimmerman EA, Tomashefsky P, Ng P-Y, Hsu KC: J Histochem Cytochem 23:208, 1975) and the "double bridge" (Ordronneau P, Petrusz P: Am J Anat 158:491, 1980) were compared at both the light and electron microscopic levels. The "double" procedures involved repeating incubations with the bridge antibody, in this case, sheep anti-rabbit gamma globulin, followed either by a second PAP step for the "double PAP" or a second anti-horseradish peroxidase step and a single incubation in horseradish peroxidase for the "double bridge." At both the light and electron microscopic levels the staining intensity was greater with the "double" techniques than with the "single" ones. This is probably due to amplification achieved with the second sheep anti-rabbit gamma globulin step, permitting an increase in the number of horseradish peroxidase molecules bound for each molecule of tissue-bound primary antibody. Also, the quality of the various commercial PAP preparations tested was variable. With the weaker ones the staining intensity could be increased by performing an incubation in fresh horseradish peroxidase after the PAP step. Finally, in electron microscopic studies, the reaction products formed in both the bridge and PAP procedures were identical in shape and size.

scholarly journals A modification of the unlabeled antibody enzyme method using heterologous antisera for the light microscopic and ultrastructural localization of insulin, glucagon and growth hormone.

1975 ◽  
Vol 23 (9) ◽  
pp. 666-677 ◽  
Author(s):  
S L Erlandsen ◽  
J A Parsons ◽  
J P Burke ◽  
J A Redick ◽  
D E Van Orden ◽  
...  
Keyword(s):  
Growth Hormone ◽  
Gamma Globulin ◽  
Cross Reactivity ◽  
Model Systems ◽  
Electron Microscopic Level ◽  
Immunocytochemical Staining ◽  
Electron Microscopic ◽  
Primary Antiserum ◽  
Enzyme Method ◽  
Unlabeled Antibody

The requirement of using homologous antisera (primary antiserum and peroxidase-antiperoxidase (PAP) complex raised in the same species) in the unlabeled antibody enzyme method has been investigated at the light and electron microscopic level using the localization of insulin, glucagon and growth hormone as model systems. Optimum immunocytochemical staining for all three antigens was observed when sheep or goat antirabbit gamma-globulin (S-ARgammaG or G-ARgammaG) were used to couple rabbit peroxidase-antiperoxidase complex with either guinea pig antisera to insulin (GP-AIS) or glucagon (GP-AGS), or monkey antisera to rat growth hormone (M-ARGH). The cross-reactivity between S-ARgammaG or G-ARgammaG and immunoglobulins in these primary antisera were substantiated by immunoelectrophoresis and radioimmunoassay. S-ARgammaG was shown to produce precipitation arcs with GP-AIS and M-ARGH that were similar to those seen when the latter were reacted with rabbit antiguinea pig gamma-globulin antiserum and goat antimonkey gamma-globulin antiserum, respectively. Radioimmunoassay results revealed that immunoprecipitation of 6-10% as compared to homologous antisera controls yielded excellent staining localization when S-ARgammaG was used for immunocytochemistry. Thus, heterologous antisera (primary antiserum and PAP complex raised in different species) may be used in the unlabeled antibody enzyme method as long as the coupling antiserum shows cross-reactivity with immunoglobulins of the primary antiserum and the PAP complex.

Ultrastructural Immunocytochemistry of Five Rat Pituitary Adenomas

1980 ◽  
Vol 38 ◽  
pp. 724-725
Author(s):  
D. J. McComb ◽  
N. Ryan ◽  
E. Horvath ◽  
K. Kovacs ◽  
E. Nagy ◽  
...  
Keyword(s):  
Pituitary Adenomas ◽  
Pituitary Tumors ◽  
Electron Microscopic ◽  
Electron Microscopic Immunocytochemistry ◽  
Transmission Electron ◽  
Rat Pituitary ◽  
Radiation Induced ◽  
Group 5 ◽  
Group 2 ◽  
Microscopic Techniques

Conventional light and electron microscopic techniques failed to clarify the cellular composition and derivation of spontaneous and induced, intrasellar and transplanted pituitary adenomas in rats (1). In the present work, electron microscopic immunocytochemistry was applied to evaluate five adenohypo-physial tumors using a technique described by Moriarty and Garner (2). Spontaneously occurring pituitary adenomas (group 1) were harvested from aging female Long-Evans rats. R-Amsterdam rats were treated with 2 x 1.0 mg estrone acetate (HogivaI) s.c. weekly for 6 months. Pituitary adenomas in excess of 30 mg were removed from these animals to make up the tumors of group 2. Groups 3 and 4 consisted of estrogen-induced autonomous transplan¬ted pituitary tumors MtT.WlO and MtT.F4. Group 5 was a radiation-induced transplanted autonomous pituitary tumor MtT.W5. The tumors of groups 3,4 and 5 were allowed to proliferate in host rats 6-8 weeks prior to removal for processing. Tissue was processed for transmission electron microscopy (glutaraldehyde fixation, OsO4 postfixation and epoxy resin embedding), and electron microscopic immunocytochemistry (3% paraformaldehyde fixation and Araldite embedding).

scholarly journals Benzene and 2-ethyl-phthalate induce proliferation in normal rat pituitary cells

Pituitary ◽  
2016 ◽  
Vol 20 (3) ◽  
pp. 311-318 ◽  
Author(s):  
Laura Tapella ◽  
Antonella Sesta ◽  
Maria Francesca Cassarino ◽  
Valentina Zunino ◽  
Maria Graziella Catalano ◽  
...  
Keyword(s):  
Pituitary Cells ◽  
Rat Pituitary ◽  
Normal Rat ◽  
Rat Pituitary Cells

Couples Facing Cancer

2021 ◽  
pp. 729-736
Author(s):  
Hoda Badr ◽  
Courtney Bitz
Keyword(s):  
Routine Care ◽  
Poor Quality ◽  
Psychosocial Adaptation ◽  
Future Research ◽  
Great Promise ◽  
Clinical Implications ◽  
Social Challenges ◽  
Healthcare Settings ◽  
Care And Support

Cancer survivors experience significant physical, psychological, and social challenges that contribute to poor quality of life. Intimate partners provide critical care and support across the cancer continuum, but they report psychological distress, lack basic healthcare knowledge and skills, and experience increased tension and conflict in their relationships with survivors. Couple-based interventions hold great promise in cancer because they can simultaneously address survivor, partner, and relationship concerns. However, they are seldom implemented in healthcare settings as part of routine care. This chapter will therefore integrate what research has taught us about couples and cancer and what we have learned from couples in the clinical setting. We begin with an overview of challenges faced by couples across the cancer continuum, including biopsychosocial stressors. Next, we describe different perspectives that have shaped descriptive and intervention research on couples’ psychosocial adaptation to cancer. We conclude with clinical implications and directions for future research.

STIMULATION OF GROWTH HORMONE RELEASE FROM SUPERFUSED RAT PITUITARY BY EXTRACTS OF HYPOTHALAMUS AND OF HUMAN LUNG TUMOURS

1973 ◽  
Vol 59 (2) ◽  
pp. 325-333 ◽  
Author(s):  
CHRISTINE BECK ◽  
R. G. LARKINS ◽  
T. J. MARTIN ◽  
H. G. BURGER
Keyword(s):  
Growth Hormone ◽  
Human Lung ◽  
Gel Filtration ◽  
Lung Tumour ◽  
Oral Glucose ◽  
Hormone Release ◽  
Lung Tumours ◽  
Rat Pituitary ◽  
Normal Rat ◽  
Hypothalamic Tissue

SUMMARY The paradoxical plasma growth hormone (GH) responses to oral glucose in certain patients with lung cancer prompted an examination of tumour extracts for GH releasing activity. Exposure of superfused rat pituitary to pulses (30 s) of aminophylline and to extracts from rat, sheep and human hypothalami resulted in a rapid and short-lived release of immunoreactive rat GH into the medium. Fresh extracts from five lung tumours, and from the surrounding lung tissue of four of these tumours significantly stimulated the release of GH, while extracts of a metastatic chondrosarcoma and normal rat lung were inactive. Gel filtration experiments suggested that the releasing activity in rat, sheep and human hypothalamic tissue and in human lung tumour extracts was present in at least two molecular species of different sizes.

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