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At least 19 recordsLinked to original sources
PubMed1977

Connective tissue activation by synovial fluids and synovial-tissue extracts of rheumatoid arthritis patients.

Prompted by Castor's investigations (4) on a connective-tissue-activating peptide (CTAP) we investigated the stimulative effect of synovial fluids and synovial-tissue extracts on the synthesis of collagen by incubated embryonic-chick tendon cells. The stimulative effect was greater with synovial-tissue extracts from more severe cases than with samples from patients having milder forms of the disease. There was no correlation between the stimulation by synovial fluids and synovial-tissue extracts from the same patient. The stimulative activity was lost at dialysis. A slight stimulation in the incorporation of glucosamine was also observed. Treatment of the patients with gold, chloroquine or steroids decreased the stimulating capacity. These effects seem to depend on factors different from those described by Castor. The differerences in the stimulant activity of samples from various groups cannot be entirely due to glutamine, which is one of the limiting nutrients of the embryonic-chick tendon cells.

Animals↗
PubMed1978

Selective inhibition of leukocyte adherence in women with breast cancer using a variety of tissue extracts.

Leukocyte adherence inhibition (LAI) tests for the assessment of tumor immunity in vitro have been based on the assumption that the adherence of the leukocytes is inhibited by specific interaction with an antigenic tumor extract. However, loss of leukocyte adherence could depend on several non-immunological factors including disease state of the leukocyte donor and the presence of a variety of agents, proteins or tissue extracts. We have studied LAI , therefore, using different species and concentrations of serum protein or tissue extracts. Our results show that leukocytes from patients with a variety of diseases adhere to glass in a rather consistent fashion. High concentrations of serum protein or tissue extract inhibited leukocyte adherence. 39 breast cancer patients and 38 female controls were studied with a total of 23 solubilized extracts from breast carcinoma, other tumor and non-tumor tissues. Breast cancer patients showed selective LAI in the presence of 200 microgram per test tube of several breast carcinoma extracts.

Antigens, Neoplasm↗
PubMed1978

[Tissue extracts of malignant human lymphomas in the leukocyte migration test].

Water-soluble lymph node extracts and sera from 23 patients with benign and malignant lymphomas and tissue extracts from liver, spleen, kidney, and muscle of one patient without any lymphatic disease were analyzed using the direct leukocyte migration test (LMT). The leukocytes were obtained from healthy persons. 10 of the lymphomas and 3 of the tissue extracts varied the leukocyte migration. In most cases of chronic nonspecific lymphadenitis, in 2 cases of Hodgkin's disease, and in extracts of liver, spleen and muscle inhibitory effects were found. Stimulating effects could be demonstrated in 2 cases of immunocytoma and in 4 cases of Hodgkin's disease. The results indicate that in cases of stimulation the target cells might be influenced by a pathological migration inhibitory factor.

Cell Migration Inhibition↗
PubMedDate not supplied

[Method of measuring binding constants and the number of specific cyclic AMP receptor sites in tissue extracts].

A method for evaluation of binding constants and of amount of cAMP binding sites in crude tissue extracts was developed. The method is based on equilibrium binding of 3H-cAMP by proteins with subsequent ultrafiltration. Hydrolysis of cAMP and its unspecific sorption by proteins were eliminated under the conditions selected. Rat spleen cytosole contained 3.57 +/- 0.34 pmol of cAMP binding sites per mg of protein with dissociation constant of protein-cAMP complex (1.68 +/- 0.28).10(-8) M. As shown by studies on kinetics, binding constants and specificity of binding, the method permitted to evaluate quantitatively cAMP-dependent protein kinases in crude tissue extracts and to estimate their affinity to cAMP.

Animals↗
PubMed1992

Homogeneous trinder-coupled assay for the determination of glucose-6-phosphatase activity in tissue extracts.

We describe an automated, homogeneous, glucose oxidase-coupled method for the determination of glucose-6-phosphatase activity in tissue extracts. The method is based on measurement of the rate of glucose formation by the Trinder reaction, in which the end product is a quinoneimine dye which absorbs maximally at 505 nm and has a molar extinction coefficient of 5700. The incubation mixture contains 20 microL of tissue extract, 25 microL of 0.5 M phosphate buffer, pH 7.0, 175 microL of Trinder/glucose-6-phosphate reagent, and 30 microL of distilled water. After a delay period of 15 min, to exhaust any glucose endogenously present in the extract, glucose production from glucose-6-phosphate is monitored at 505 nm for 5 min in a centrifugal analyzer. The Km was 13 mM over a 10-fold range in glucose-6-phosphate concentration and the reaction was linear up to about 250 U/L. Within-run CV of the assay at activities of 48 and 190 U/L ranged between 2.5-5.0%. The between-run CV at 190 U/L was 5.1%.

Animals↗
PubMed1976

Absence of collagen stimulating factors for L929 cells in granulation tissue extracts.

We have attempted to find collagen synthesis stimulating activity in polyvinyl sponge granulation tissue extracts and inflammatory fluid. The cells studied in culture were mouse L929 cells grown in monolayers. A prior study by another group reported such factors to be present in healing wound extracts and in extracts of CHHl3 injured liver. We have been unable to find such stimulating activity for L929 cells in granuloma tissue fluid. In addition we have examined the procedures used by the prior group and have found major problems in their methodology which are discussed below.

Animals↗
PubMedDate not supplied

Muscle trophic factor: III. Effect of hormones and tissue extracts on muscle trophic-factor activity.

The ability of classical hormones or extract from adult chicken tissues to replace or influence the activity of chicken skeletal muscle cell trophic factor was investigated. Pituitary gland extract did not replace the trophic factor in a range equivalent to the serum concentrations of somatotropin, although high concentrations of such extract showed a significant ability to mimic trophic-factor activity. Insulin, triiodothyronine, testosterone, dihydrotestosterone (Stanolone), and estradiol did not show any ability to mimic trophic-factor activity. Both pituitary extract and insulin showed a potentiating effect on the trophic factor when added to the medium; however, the concentrations necessary for this effect were too high to be considered physiologic. Triiodothyronine, testosterone, Stanolone, and estradiol did not potentiate the trophic factor; indeed, at higher concentrations, these hormones actually suppressed the activity of the trophic factor. Every tissue extract examined showed, to a greater or lesser extent, the ability to potentiate the trophic factor.

Animals↗
PubMed1977

A neuromuscular transmission block produced by a cancer tissue extract derived from a patient with the myasthenic syndrome.

The myasthenic syndrome occasionally is associated with bronchogenic carcinoma. The neuromuscular transmission defect in this syndrome is characterized by a reduction of acetylcholine release from motor nerve endings. This paper reports that an acetone extract of cancer tissue from a patient with the syndrome reduces the acetylcholine release from motor nerve endings and produces a neuromuscular transmission defect in the frog nerve-muscle preparation. This suggests that the pathogenic substance(s) contained in the extract may be produced by certain types of bronchogenic carcinoma and may cause the myasthenic syndrome.

Acetylcholine↗
PubMed2025

Systematic comparison of quantity and quality of RNA recovered with commercial FFPE tissue extraction kits.

BACKGROUND: FFPE tissue samples are commonly used in biomedical research and are a valuable source for next-generation sequencing in oncology, however, extracting RNA from these samples can be difficult the quantity and quality achieved can impact the downstream analysis. This study compared the effectiveness of seven different commercially available RNA extraction kits specifically designed for use with FFPE samples in terms of the quantity and quality of RNA recovered. METHODS: This study used 9 samples of FFPE tissue from three different types of tissue (Tonsil, Appendix and lymph node of B-cell lymphoma) to evaluate RNA extraction methods. Three sections of 20 µm of each sample were combined per sample. The slices were distributed in a systematic manner to prevent any biases. Each of the 7 commercially available RNA extraction kits were used according to manufacturer's instructions, with each sample being tested in triplicate resulting in a total of 189 extractions. The concentration, RNA quality score (RQS) and DV200 of each extraction was analysed using a nucleic acid analyser to determine the quantity and quality of the recovered RNA. RESULTS: This study found that despite processing the FFPE samples in the same standardized way, there were disparities in the quantity and quality of RNA recovered across the different tissue types. Additionally, the study found notable differences in the quantity of RNA recovered when using different extraction kits. In terms of quality, three of the kits performed better than the other four in terms of RQS and DV200 values. CONCLUSION: Though many laboratories have developed their own protocols for specific tissue types, using commercially available kits is still a popular option. Although these kits use similar processes and extraction procedures, the amount and quality of RNA obtained can vary greatly between kits. In this study, among the kits tested, while the Roche kit, provided a nearly systematic better-quality recovery than other kits, the ReliaPrep FFPE Total RNA miniprep from Promega yielded the best ratio of both quantity and quality on the tested tissue samples.

Paraffin Embedding↗
PubMed1978

The nature of the immunoreactive lipotropins in human plasma and tissue extracts.

This study was designed to establish definitively the nature of immunoreactive lipotropin (IR-LPH) in human plasma and tissue extracts. Using gel filtration, gel filtration under denaturing conditions, cationic exchange chromatography, immunoprecipitation, and radioimmunoassay, we have studied normal and tumorous human pituitaries, ectopic ACTH- and LPH-secreting tumors, plasma from normal subjects before and after dexamethasone administration, and plasma from patients with primary adrenal insufficiency and pituitary and nonpituitary ACTH- and LPH-secreting tumors. Except in the plasma and tumors of occasional patients with ectopic ACTH syndrome, the smallest IR-LPH appears to be lambda-lipotropin (lambdaLPH), which is often the predominant and occasionally the only IR-LPH present. The other major peptide appears to be betaLPH, a 91-amino acid molecule that contains lambdaLPH as its 1-58 sequence. Larger immunoreactive materials were observed in some specimens, but the "big" LPH in one plasma was shown to be lambdaLPH bound to IgG.The weak melanocyte-stimulating activity of LPH suggests that ACTH may be the principal pigmentary hormone in man. The fact that lambdaLPH, rather than betaLPH, is the predominant form in plasma suggests that the enkephalin-endorphin opiate peptides, which are contained in the "missing" 59-91 sequence from the betaLPH precursor of lambdaLPH, may be secreted in parallel with ACTH under both physiological and pathological conditions in man.

Chromatography, Gel↗
PubMed1976

Determination of protein in adipose tissue extracts.

An accurate and sensitive method for determination of protein in adipose tissue extracts based on a scaled-down version of the method of Lowry et al. (1951, J. Biol. Chem. 193, 265-275) is described. Protein is quantitatively precipitated by trichloroacetic acid with a nonionic detergent as a bulk-increasing substance and the precipitate is extracted with an organic solvent. This interference caused by lipids, sulfhydryl reagents, and several other compounds is eliminated. The accuracy of the determination of dilute reference protein solutions (2-20 mug/ml) is demonstrated.

Adipose Tissue↗
PubMed1992

Improved and simplified tissue extraction method for quantitating long-chain acyl-coenzyme A thioesters with picomolar detection using high-performance liquid chromatography.

A method has been developed that permits rapid and easy tissue extraction of long-chain acyl-coenzyme A (acyl-CoA) thioesters with sensitive quantitation by reversed-phase high-performance liquid chromatography (RP-HPLC). Tissue homogenants are extracted using a reserve Bligh-Dyer technique, and long-chain acyl-CoA esters are harvested in the methanolic aqueous phase. Complex lipids and phospholipids are removed in the chloroform-rich organic Bligh-Dyer second phase, and long-chain acyl-CoA compounds are further purified from the methanolic aqueous Bligh-Dyer first phase on C18 extraction columns after removal of the methanol. The eluted and purified acyl-CoA esters are then quantitated by RP-HPLC using heptadecanoyl-CoA as an internal standard resulting in a detector sensitivity of about 12 pmol. Ten long-chain acyl-CoA esters from C12:0 to C20:4 were identified and separated from canine renal cortex and murine liver samples. The predominant acyl-CoA peaks from both kidney and liver were 14:0, 16:1, 16:0, 18:1, 18:2 and 20:4. Murine liver also produced 18:0 and all peaks disappeared after alkaline hydrolysis of the samples. This extraction and quantitation technique can successfully be used for tissue samples as small as 20 mg, and many samples can be processed in a short period of time. The simplicity of the extraction procedure and the sensitivity of the assay make this an attractive alternative approach to quantitating long-chain acyl-CoA thioesters from complex biological samples such as tissues.

Acyl Coenzyme A↗
PubMed1979

Morphologic study of the action of lymphoid tissue extracts in peroxidase arthritis.

By means of modelling arthritis by horseradish peroxidase it was possible to reproduce a process corresponding to immune synovitis. A considerable reduction of immunomorphological phenomena in joints, lymphoid tissue, heart, liver and kidneys was favoured by application of lymphoid tissue extracts. Comparison between the extracts of spleen and lymph nodes showed a more pronounced effect of spleen extract.

Animals↗
PubMed1977

Specific restoration of delayed hypersensitivity by lymphoid tissue extracts.

Mice lose demonstrable delayed hypersensitivity (DH) to DNFB, picryl chloride, or sheep red blood cells. Reconstitution of immune responsiveness can be accomplished by administration of cell-free lysates of spleens from mice with active DH to structurally related, but not to unrelated antigens. Peritoneal exudate cell lysates from mice with active DNFB-DH also restore DH to this antigen. Sera from sensitized mice, and sera and lymphoid tissue extracts from unsensitized mice are without activity. The restorative property of splenic lysates from DNFB-sensitized mice is unstable at 56 degrees C, not sedimented at 90,000 X G and inactivated by trypsin or magnesium ions. The presence of unexpressed, restorable DH may provide a biologic basis for the so called "transfer factor" phenomenon.

Animals↗
PubMed1975

Microbial adjuvant and autoimmunity. I. Induction of antibody responses to syngeneic tissue extracts in mice treated with capsular polysaccharide of Klebsiella pneumoniae.

With the use of the capsular polysaccharide of Klebsiella pneumoniae (CPS-K) as a powerful adjuvant, high precipitin responses could be induced in mice to syngeneic eyeball extracts and thyroid gland extracts which were normally nonimmunogenic. Only very weak responses were induced to eyeball extracts by Freund's complete adjuvant. Repeated administrations of the antigens mixed with CPS-K at time intervals of 30 days (more than twice for the eyeballs or more than three times for the thyroid glands) were required for induction of high precipitin responses. Antibody responses detectable by the immunofluorescent technique could be induced to syngeneic lymphoid tissue extracts by injecting the mixture of antigen and CPS-K more than five times at time intervals of 30 days. These findings suggest that repeated stimulation by autoantigens together with such a strong adjuvant as CPS-K can terminate natural tolerance against autoantigens.

Adjuvants, Immunologic↗
PubMed1975

Carcinoembryonic antigen (CEA) of human tissue extracts: partial characterization of two variants separated by affinity chromatography on concanavalin A.

The carcinoembryonic antigen (CEA) of normal and pathological tissue extracts was separated into two variants, Concanavalin A reactive (CEAr) and non-reactive CEA (CEAn) by affinity chromatography on Con A Sepharose columns. CEAr was the quantitatively predominant variant. CEAn varied in concentration between 0.2 and 6 percent of the total CEA activity. The affinity of CEAn for anti-CEA antibodies was significantly lower than that of CEAr. Pooled extracts of primary adenocarcinomas of the colon contained CEAn in the lowest concentration and with the least affinity for antibodies. It is suggested that a deficiency and/or steric blocking of alpha-D-mannopyranosyl residues in CEAn reduce the affinities for both antibodies and Con A.

Adenocarcinoma↗
PubMed1975

[The diangostic value of skin tests with tissue-extracts, especially in lepra and sarcoidosis (author's transl)].

The immunological aspects of lepra (India, Germany) and Sarcoidosis are presented under consideration of cutaneous reaction to lepromin and Kveim-extracts. Patients with lepra, erythema nodosum, sarcoidosis and different dermatoses have been tested with lepromin, Kveim-Antigen and tuberculin. The specifity of the results has been discussed. Patients with tuberculoid lepra did not react on Kveim-test, but patients with sarcoidosis can give positive reaction on lepromin. Patients with erythema nodosum without clinical signs of sarcoidosis are Kveim negativ and a positive lepromin-test is an exception. The positive Kveim-test is bound to a histological examination. The positive lepromin-test in non lepraendemic countries should be interpreted also by histological examination. Granulomatous dermatoses can show positive lepromin-tests, but the histological picture is different from the lepromin reaction in tuberculoid lepra. The positive reactions on tissue extracts are not combined with positive immunfluorescence results. Intracutaneous injection of lepra-bacilli in lepromin extract can be presented days to weeks after intracutaneous application in biopsies.

Antigens↗
PubMed1976

The reduction of oxo-acids by human tissue extracts.

The reduction of pyruvate, 2-oxobutyrate, hydroxy-pyruvate and glyoxylate under optimal conditions at 37 degrees by extracts of human heart, kidney, pancreas, skeletal muscle and liver has been examined. The ratios of reduction rates, particularly of glyoxylate and 2-oxobutyrate reveal a capacity to differentiate the isoenzyme composition of the tissue extracts.

Humans↗