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At least 163 records · Page 9Linked to original sources
PubMed1977

Mitotic segregation of cytoplasmic determinants for chloramphenicol resistance in mammalian cells. I: Fusion with mouse cell lines.

The segregation of cytoplasmically inherited chloramphenicol (CAP) resistance in mouse cells was investigated in fusions between CAP-resistant cells or cytoplasts (enucleated cells) and CAP-sensitive cells of varying tissue origin. All hybrids formed in cell-cell fusions were initially CAP-resistant, indicating that CAP resistance is dominant. Hybrids from fusions of cells of the same tissue origin (homologous) were stably CAP-resistant, whereas the hybrid population from fusions of different origins (heterologous) showed a rapid diminution of average CAP resistance. Individual hybrid clones from these heterologous fusions also showed an overall loss of CAP resistance, and a wide variation in CAP resistance which is consistent with a large number of genetic determinants (possibly mitochondrial DNA molecules) contributing to the CAP phenotype. Similar results were obtained from cytoplast-cell fusions, so the observed CAP segregation is not the result of nuclear-nuclear interactions. This segregation of CAP resistance constitutes a second criterion of cytoplasmic inheritance in mammalian cells.

Animals↗
PubMed1985

The steroid hormone receptors in tumors of adipose tissue.

The clinical evidence suggests that the steroid hormones may influence the biologic course of tumors of soft tissue. In an attempt to characterize the possible steroid hormone dependency of soft tissue, we studied the incidence and distribution of cytosolic receptors for the steroid hormones in benign and malignant tumors of adipose tissue origin. All specimens were assayed for the steroid hormone receptors by charcoal dextran technique and analyzed by the method of Scatchard. The results show a high incidence of cytosolic estrogen and glucocorticoid receptors in tumors of adipose tissue origin except in lipoma and well differentiated liposarcoma. The binding parameter of these receptors suggests that the receptors may be responsive to physiologic steroid hormonal milieu.

Adipose Tissue↗
PubMed1978

Initial clinical evaluation of a new Rocky Mountain spotted fever vaccine of tissue culture origin.

Currently available Rocky Mountain spotted fever (RMSF) vaccines are relatively ineffective in preventing infections in humans and contain considerable amounts of contaminating egg protein. A new formalin-inactivated vaccine was prepared by sucrose density gradient centrifugation of the Sheila Smith strain of Rickettsia rickettsii grown in chick embryo cell tissue culture. The new product has greater protective immunogenicity in rheusus monkeys and guinea pigs than commercial vaccines. Six volunteers without immunologic evidence of prior exposure to RMSF received from one to three inoculations of the vaccine diluted 1:10, and there were two benign local reactions. Titers of antibody (determined by microagglutination and indirect fluorescence techniques) increased in all recipients as did lymphocyte tranformation responses to specific rickettsial antigen. Ten volunteers were immunized twice with vaccine diluted 1:3; there were no local reactions, and immunologic responses were similar to those in the six volunteers in the first group. The proper dosage and immunization schedule for the vaccine must be determined in further studies.

Agglutinins↗
PubMed1992

Effects of culture passages on collagen immunostaining in human dental pulp and gingival fibroblasts.

In this study, a scanning microscopic computer-assisted image analysis system was used for the immunocytochemical characterisation of collagen types I, III and V in normal human fibroblasts from pulp and gingival explants, using specific purified antibodies and peroxidase labeling. The culture conditions were standardized in order to evaluate simultaneously the expression of the three antigens in four different culture passages of the two fibroblast types. The optical density values of immunostaining intensities were quantified, the integrated optical density per cell was calculated, and the results were analyzed by a variance test. It was found that all three collagen types were present in the tissues, and in both gingival and pulp fibroblasts after three to nine culture passages. A non-parametric statistical analysis of the staining intensity variances revealed significant differences between antigenic levels depending on the tissue origin of the fibroblasts and an effect of culture passages. The results seemed to justify application of this technique at the light microscope level for the evaluation of collagen production, the principal function of fibroblasts, but the tissue origin and number of culture passages should be taken into consideration for in vitro biocompatibility testing of dental materials.

Adult↗
PubMed1976

Ethylenimine-inactivated rabies vaccine of tissue culture origin.

The replication of seven rabies virus strains (CVS, HEP, PV, ERA, WIRAB, CPZ and BOLIVAR) in BHK cells and the inactivation dynamics of these strains by beta-propiolactone, acetylethylenimine, and ethylenimine were studied to find the most immunogenic strain and the most economic and stable inactivating agent for the production of an inactivated tissue culture rabies vaccine for animal use. The seven strains reached the peak of virus production 3 to 5 days after inoculation of the cell culture; PV yielded the highest virus titer (10(9) plaque-forming units/ml). The infectivity of virus suspensions containing 10(7) to 10(8) plaque-forming units/0.1 ml was inactivated by beta-propiolactone in 0.5 h, acetylethylenimine in 3.0 h, and ethylenimine in 1.0 h. Most of the vaccine lots prepared with the different strains and inactivating agents passed a modified National Institutes of Health potency test. The vaccines prepared with the PV strain had consistently higher antigenic values (equal or better than four) than the other six strains. This difference was highly significant (F6,12=59.8), whereas there were no statistically significant differences among the antigenic values of the vaccine lots prepared with the three inactivating agents. Batches of lyophilized and liquid vaccine stored at 4 C maintained potency for over 1 year. Ten dogs vaccinated with a vaccine prepared with the PV strain and inactivated with ethylenimine developed a good antibody response and resisted challenge 60 days after vaccination, while seven of eight nonvaccinated controls died of rabies. This information indicates that an inactivated, stable, economic, and easy-to-prepare rabies vaccine can be produced in BHK cells by using the PV strain and ethylenimine as an inactivating agent.

Animals↗
PubMed1995

[A comparative study of the synthesis of membrane hetero-organic antigens of kidney origin on tissue slices and isolated cells after a single carcinogenic exposure in rats].

A comparative study was carried out of a single carcinogenic injection of rats with a "weak" carcinogen, 4-dimethylaminoazobenzene (DAB), and a "strong" carcinogen, N-diethylnitrozamine (DENA). Both carcinogenic agents caused similar antigenic rearrangements involving the appearance of membrane hetero-organic antigens of kidney origin associated with the Zajdela hepatoma on the hepatocyte membrane. The expression of these antigens is longer in DENA carcinogenesis. On the sections of embryonic liver (16-18 days) single hepatocytes were discovered carrying membrane hetero-organic antigens. No synthesis of these antigens was discovered in cells of the definitive liver, but it was seen to resume at early stages of carcinogenesis. Quantitative characteristics of synthesis of the investigated antigens was done by means of immunocytofluorometric method using membranes of isolated hepatoma cells and hepatocytes of rats after a single carcinogenic injection and partial hepatectomy.

Animals↗
PubMed1982

Hormone-sensitive lipase in human adipose tissue, isolated adipocytes, and cultured adipocytes.

Employing a perifusion technique, the activity of hormone-sensitive lipase (HSL) in human adipose tissue and isolated adipocytes was found to be significantly stimulated by 5 microM epinephrine (5-fold. P less than 0.001) or 0.5 mM theophylline (2-fold, P less than 0.03). The stimulatory effect of both, however, was not cumulative; instead, theophylline appeared to blunt the epinephrine effect. The two effectors together produced a 3-fold increase in activity over basal (P less than 0.01). Basal lipolysis was highly and significantly correlated with epinephrine and/or theophylline stimulated lipolysis in the tissue as well as in the isolated cells. The activity of the enzyme was examined in cultured human adipocytes using a cell-free system. The basal activity of HSL in the 20,000 X g supernatant (S20) fraction of cultured cells grown in fat-enriched medium was significantly higher than the value in the same fraction of cells grown in regular medium (6.06 +/- 1.49 versus 2.78 +/- 0.89 nmole glycerol/min/mg protein, mean +/- S.D., P less than 0.01), an was similar in the S20 fractions of cells grown in the enriched medium and the original tissue (6.06 +/- 1.49 versus 5.44 +/- 2.73 nmole glycerol/Min/mg protein, mean +/- S.D., P greater than 0.83). When the tissue and cells were stimulated in vitro before fractionation, the HSL activity in the S20 fraction of the original tissue increased 4-fold over basal (P less than 0.001), whereas that in the cultured cells increased 2-fold (P less than 0.01) regardless of the culture medium employed. In the S20 fractions derived from basally incubated samples, but not in those derived from the epinephrine-stimulated samples, the HSL enzyme was slightly but significantly (P less than 0.002) activated by exogenous addition of ATP, cyclic AMP, and protein kinase. The data suggest that the activity of HSL is retained in cultured human adipocytes and could be enhanced under conditions of culture which favor lipid accumulation and adipose conversion.

Adipose Tissue↗
PubMedDate not supplied

Derivation of the mammalian skull vault.

This review describes the evolutionary history of the mammalian skull vault as a basis for understanding its complex structure. Current information on the developmental tissue origins of the skull vault bones (mesoderm and neural crest) is assessed for mammals and other tetrapods. This information is discussed in the context of evolutionary changes in the proportions of the skull vault bones at the sarcopterygian-tetrapod transition. The dual tissue origin of the skull vault is considered in relation to the molecular mechanisms underlying osteogenic cell proliferation and differentiation in the sutural growth centres and in the proportionate contributions of different sutures to skull growth.

Animals↗
PubMed2002

Distinct central representations of inescapable and escapable pain: observations and speculation.

It is well established clinically that the affective response to pain of deep origin (muscles, joints and viscera) is distinct from that evoked by cutaneous pain. Cutaneous pain triggers a fight-flight reaction (active emotional coping), whereas deep pain evokes a reaction of quiescence, decreased vigilance and vasodepression (passive emotional coping). These observations led to suggestions of distinct central representations for deep versus cutaneous pain. Indeed, studies using immediate early gene (c-fos) expression revealed selective activation of ventrolateral versus lateral columns of the midbrain periaqueductal grey region (PAG) by persistent pain of deep origin versus intermittent cutaneous pain. Ventrolateral versus lateral PAG activation had been found earlier to evoke passive versus active emotional coping. However, not all cutaneous pain triggers active coping. Persistent cutaneous pain (e.g. burns) instead, usually evokes passive coping. This raised the question of whether the behavioural significance of pain (i.e. its escapability versus inescapability), rather than its tissue origin, is represented in supraspinal regions such as the PAG. Subsequent study revealed that a persistent (inescapable) noxious cutaneous manipulation (clip of the neck) evoked both selective ventrolateral PAG Fos expression and passive emotional coping. Such data suggest that pain representation in the PAG reflects a quality akin to behavioural significance, rather than tissue origin. In contrast, in the spinal cord predominantly superficial dorsal horn Fos expression was seen after either persistent or intermittent noxious cutaneous stimuli, leaving the question of the pathway(s) via which persistent (inescapable) cutaneous pain activates the vlPAG unanswered. One experimental approach to this question is suggested.

Animals↗
PubMedDate not supplied

[Immunohistochemical study of blood group activities in severely burned human tissue].

The blood, livers and lungs obtained from donors or cadavers of known blood groups were experimentally burned, while the temperatures inside the tissue specimens were automatically measured. All the distinguishable portions with various thermo-changes in each burned tissue specimen were examined for their blood-group activities A, B, Lea, Leb and P1 by means of the immunofluorescence and immunoperoxidase techniques. In the layer immediately before charring of the blood masses (tissue temperature: ca. 200-250 degrees C), the activities A, B, Lea, Leb and P1 could be specifically demonstrated on the erythrocyte membranes. In case of the burned livers and lungs, the A- and B-activities could be detected in the small blood vessels in the layer immediately before charring (ca. 200-250 degrees C), though the layer was so severely thermo-changed, that their original tissue structures could be no more observed. In the severely thermo-coagulated, porous and hardened layer of both organ tissues, the A- and B-activities could be demonstrated on the endothelial cells of the hepatic sinus and small blood vessels (especially in the alveolar walls). The simply thermo-coagulated inner portions well retained their original tissue structures and the A- and B-activities remained on the epithelial cells of the alveoli and bronchioles of the lung as well as the cells described above. The Lewis blood-group activities could be demonstrated on the epithelial cells of the bronchioles in the simply thermo-coagulated inner portion of the lung. The P1-activity could not be demonstrated in the burned tissues of the livers and lungs.

Blood Grouping and Crossmatching↗
PubMed2006

Characterization of novel elongated Parvulin isoforms that are ubiquitously expressed in human tissues and originate from alternative transcription initiation.

BACKGROUND: The peptidyl prolyl cis/trans isomerase (PPIase) Parvulin (Par14/PIN4) is highly conserved in all metazoans and is assumed to play a role in cell cycle progression and chromatin remodeling. It is predominantly localized to the nucleus and binds to chromosomal DNA as well as bent oligonucleotides in vitro. RESULTS: In this study we confirm by RT-PCR the existence of a longer Parvulin isoform expressed in all tissues examined so far. This isoform contains a 5' extension including a 75 bp extended open reading frame with two coupled SNPs leading to amino acid substitutions Q16R and R18S. About 1% of all Parvulin mRNAs include the novel extension as quantified by real-time PCR. The human Parvulin promoter is TATA-less and situated in a CpG island typical for house keeping genes. Thus, different Parvulin mRNAs seem to arise by alternative transcription initiation. N-terminally extended Parvulin is protected from rapid proteinaseK degradation. In HeLa and HepG2 cell lysates two protein species of about 17 and 28 KDa are detected by an antibody against an epitope within the N-terminal extension. These two bands are also recognized by an antibody towards the PPIase domain of Parvulin. The longer Parvulin protein is encoded by the human genome but absent from rodent, bovine and non-mammalian genomes. CONCLUSION: Due to its molecular weight of 16.6 KDa we denote the novel Parvulin isoform as Par17 following the E. coli Par10 and human Par14 nomenclature. The N-terminal elongation of Par17-QR and Par17-RS suggests these isoforms to perform divergent functions within the eukaryotic cell than the well characterized Par14.

Amino Acid Sequence↗
PubMed2005

Establishment of subrenal capsule xenografts of primary human ovarian tumors in SCID mice: potential models.

OBJECTIVE: To evaluate subrenal capsule xenografting of primary ovarian tumor tissues in mice for development of new ovarian cancer models. METHODS: Pieces (1 x 3 x 3 mm) of ovarian tumor specimens from patients were meticulously grafted under renal capsules of female NOD/SCID mice within 2 h of surgical removal. Tumor types included papillary serous adenocarcinomas, borderline and benign mucinous cystadenomas, granulosa cell tumors, a serous borderline tumor and a grade 3 mixed surface epithelial tumor of transitional and undifferentiated types. After 1-2 months, grafts were retrieved for comparison with original tissues. Hematoxylin and eosin (H&E) and immunohistochemical staining was carried out using tissue micro-arrays and CEA, B72.3, WT-1, OC125, keratin, inhibin, CK7, CK20, Cam5.2, and MIB-1 as markers. RESULTS: Tumor tissue engraftment rate was > 95%. Comparison of donor and post-graft tissues showed highly similar histopathological features; 91 +/- 5% concordance in immunostaining indicated major preservation of immunophenotypes in the xenografts for 30-60 days. There was a small, but significant, increase in MIB-1 proliferative index in xenografts compared to original specimens. CONCLUSIONS: Subrenal capsule xenografts of primary human ovarian tumors in SCID mice can retain major histopathological and immunohistochemical characteristics of the original tissues. The achievable, consistently high engraftment rate allows use of such xenografts as tools for studying a wide range of ovarian tumors, including granulosa cell tumors and benign, borderline, and malignant surface epithelial neoplasms. Potential applications include preclinical testing of patients' tumor responses to various chemotherapeutic regimens, evaluation of novel therapeutic agents, analysis of tumor progression at cellular and molecular levels, and identification of new therapeutic targets.

Animals↗
PubMed1985

Suppression of cellular immune reactions by rat ovarian cells.

The influence on the immunologic response by rat ovarian cells of follicle and stromal tissue origin was investigated using the mixed lymphocyte reaction (MLR), the phytohemagglutinin (PHA) assay, the cytotoxic T lymphocyte (CTL) assay, and a cytotoxic T-cell-mediated microcytotoxicity test. The MLR between BN/Mai Pfd (RT-In) stimulator splenocytes (mitomycin-C treated) and R/A Pfd (RT-1u) responder lymph node cells was markedly suppressed by ovarian cells of follicle and/or stromal tissue origin. A similar in vitro inhibition was observed with cell-free supernatants from ovarian cells, not only in the rat MLR but also in the mouse MLR between Balb/c stimulator splenocytes and C57B1 responder lymph node cells. Moreover, the reactivity of rat lymphocytes was suppressed when they were cocultured with ovarian cells in the PHA and CTL assays. Preincubation of the supernatants with serial diluted antiprogesterone serum (APS) and antiestradiol serum (AES) revealed that the suppressive effect of these supernatants could be completely abolished with APS--and, to a lesser extent, by AES. As shown by the cell-mediated microcytoxicity assay both the ovarian cells and the steroids secreted by these cells were, however, ineffective in suppressing the effector phase. It is suggested that the steroid secretion by the ovary may play an important role in the prolonged survival of ovarian grafts.

Animals↗
PubMed1983

Skin-associated lymphoid tissues (SALT): origins and functions.

The skin has an unusual set of immunologic requirements. It is confronted by a specialized set of pathogenic organisms and environmental chemicals that represent a distinctive spectrum of antigenic specificities. Skin is subjected to physicochemical stresses such as irradiation with ultraviolet light that alter dramatically its immunologic properties. It is proposed that nature has provided skin with a unique collection of lymphoid cells, reticular cells, and organized lymphoid organs to deal with these special demands. Evidence in favor of the existence of skin-associated lymphoid tissues (SALT) includes (1) the cutaneous microenvironment is capable on its own of accepting, processing, and presenting nominal antigen; (2) strategically located peripheral lymph nodes are able to accept immunogenic signals derived from skin; (3) subsets of T lymphocytes display differential affinity for skin and its associated peripheral nodes; and (4) acquisition of this affinity by T cells is determined at least in part by differentiation signals received in situ from resident cutaneous cells. Responsibility for the establishment and integration of SALT rests with keratinocytes, Langerhans cells, and immunocompetent lymphocytes, each of which contributes uniquely to the synthesis. Together they provide skin with immune surveillance that effectively prejudices against the development of cutaneous neoplasms and persistent infection with intracellular pathogens. In patients who have been under long-term immunosuppressive therapy, the large majority of nonlymphoid malignancies arise within the skin, rather than other types of tissues. These data suggest that immune surveillance, once thought to be an immune defense operative in all somatic tissues, is a specialized immune function dedicated to the skin and mediated by SALT.

Animals↗
PubMed2000

Immortalization of bovine dental papilla cells with simian virus 40 large t antigen.

Primary cultures of dental papilla-derived cells have a limited lifespan in vitro and can be maintained only up to passage 7-9 before showing senescence, but in vitro investigations often require a large number of cells showing phenotypic characteristics of the original tissue. To overcome this shortcoming, second-passage cells established from calf molar tooth germs by enzymatic pretreatment of the dental papilla were transfected by electroporation with pSV3neo, coding for the oncogene simian virus 40 large t antigen and a neomycin-resistance gene. Under selection by G418 (neomycin), four cell clones were isolated by single cell dilution at passage 15. Integration of simian virus 40 large t antigen and expression of the gene products were determined in cell clones by polymerase chain reaction (PCR) and immunohistochemistry. Four transfected cell lines (clones B, C, D and no. 12) were maintained in culture for over 1.5 years. For cell characterization, gene expression of procollagen alpha1 (I) and osteocalcin was evaluated by reverse transcriptase (RT)-PCR with cDNA obtained from the established cell lines at passage 20. Expression of collagen type I, osteocalcin and dentine phosphoprotein was evaluated immunohistochemically at passage 20 and after 1.5 years of continuous cell culture. Gene expression and the expression of mineralized tissue-specific proteins was demonstrated with RT-PCR and immunohistochemistry within all four immortalized cell lines. Expression of dentine phosphoprotein was observed in three simian virus 40 large t antigen-transfected cell lines, suggesting the immortalization of odontoblast-like cells in vitro. Thus, transfection of bovine dental papilla-derived cells resulted in immortal cell lines exhibiting phenotypic characteristics of the original tissue.

Animals↗
PubMed2005

Manual exfoliation of fresh tissue obviates the need for frozen sections for molecular profiling.

BACKGROUND: Simple, rapid tissue processing that preserves macromolecules will enhance translational research capabilities. Traditional fixative-based approaches for specimen preservation are ideal for histologic evaluation but are not conducive to molecular studies of nucleic acids and protein. Tissue cryosections preserve macromolecule integrity, but the process is labor intensive and technically challenging. To the authors' knowledge to date, an alternative method capable of retrieving cells while providing adequate histologic detail yet preserving macromolecule integrity has been lacking. In the current study, the authors evaluated the utility of using manual exfoliation of clinical tissue samples as a means of obtaining cells for molecular analysis. This technique possesses the advantages of fixed and frozen tissue sections without their drawbacks. This simple, rapid, nonfixative based technique is capable of preparing cells from human clinical material for further isolation without compromising the preservation of macromolecules in the tissue. METHODS: Cells from a variety of clinical resection specimens from solid tumors were directly scraped from the tissue samples using the edge of a glass microscope slide and smeared onto another slide for cytologic evaluation. The manually exfoliated cells were evaluated microscopically for cytologic quality and cellular quantity. Pure cell populations were procured by laser capture microdissection (LCM) with subsequent extraction of nucleic acids and proteins. The integrity and suitability of the recovered nucleic acids and proteins for molecular analysis were evaluated using the polymerase chain reaction (PCR), reverse transcriptase-PCR, and reverse-phase protein microarray, respectively. RESULTS: Manual exfoliation permits the selection of homogeneous cell populations by LCM based on well established cytologic characteristics. DNA and mRNA, of comparable quality to frozen sections, can be amplified from the manual exfoliation cells. Proteins of similar quality can be recovered using this technique and quantitated via reverse-phase protein microarray. CONCLUSIONS: Molecular macromolecules of high quality and sufficient quantity can be retrieved from human clinical samples using manual exfoliation and LCM to procure specific cell populations. The manual exfoliation technique does not destroy the original tissue source, thereby allowing subsequent formalin tissue fixation. The technique of manual exfoliation in conjunction with LCM can enable the molecular profiling of a sampled selected cell population. Because it does not destroy the original tissue, histologic correlation can be combined with molecular profiling.

Cell Separation↗
PubMed2001

Risk analysis: divergent models and convergent interpretations.

Material presented at a NASA-sponsored workshop on risk models for exposure conditions relevant to prolonged space flight are described in this paper. Analyses used mortality data from experiments conducted at Argonne National Laboratory on the long-term effects of external whole-body irradiation on B6CF1 mice by 60Co gamma rays and fission neutrons delivered as a single exposure or protracted over either 24 or 60 once-weekly exposures. The maximum dose considered was restricted to 1 Gy for neutrons and 10 Gy for gamma rays. Proportional hazard models were used to investigate the shape of the dose response at these lower doses for deaths caused by solid-tissue tumors and tumors of either connective or epithelial tissue origin. For protracted exposures, a significant mortality effect was detected at a neutron dose of 14 cGy and a gamma-ray dose of 3 Gy. For single exposures, radiation-induced mortality for neutrons also occurred within the range of 10-20 cGy, but dropped to 86 cGy for gamma rays. Plots of risk relative to control estimated for each observed dose gave a visual impression of nonlinearity for both neutrons and gamma rays. At least for solid-tissue tumors, male and female mortality was nearly identical for gamma-ray exposures, but mortality risks for females were higher than for males for neutron exposures. As expected, protracting the gamma-ray dose reduced mortality risks. Although curvature consistent with that observed visually could be detected by a model parameterized to detect curvature, a relative risk term containing only a simple term for total dose was usually sufficient to describe the dose response. Although detectable mortality for the three pathology end points considered typically occurred at the same level of dose, the highest risks were almost always associated with deaths caused by tumors of epithelial tissue origin.

Aerospace Medicine↗
PubMed1985

A microassay for the determination of binding parameters of estrogen and androgen receptors employing affinity immobilization on Cibacron blue 3GA-Sepharose 6B.

The problems of currently available ligand-binding assays for sex-steroid receptor proteins include the relatively large mass of tissue required, the interference by sex hormone-binding globulin (SHBG), and use in the androgen receptor (AR) assay of the unstable synthetic ligand methyltrienolone. To overcome these difficulties the stabilizing effect of the dye Cibacron blue 3GA on AR and estrogen receptor (ER) proteins, and its ability to bind to these proteins, was utilized in developing an assay system for each receptor that could be applied to small samples. Use of the affinity gel Cibacron blue 3GA-Sepharose 6B (Blue gel) for the immobilization of AR, ER, and the steroid ligands bound to these receptors in the standard two-tier column assay system enabled the use of a 1:100 (original tissue weight:volume) concentration, making possible full (5-7 point) Scatchard analysis on tissue specimens of a mass as low as 15-20 mg. Significant stabilization of AR and ER was observed and association constants for these receptors were of a similar order of magnitude to those obtained either by Sephadex LH-20 gel filtration or the dextran-coated charcoal adsorption technique. Inactivation by dilution was shown to be largely prevented based on results obtained with cytosol concentrations from 1:5 to 1:100 (original tissue weight:volume). Because Blue gel does not bind SHBG, the natural steroid 5 alpha-androstan-17 beta-ol-3-one (DHT) may be employed as a ligand in the AR assay.(ABSTRACT TRUNCATED AT 250 WORDS)

Adsorption↗