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At least 127 records · Page 7Linked to original sources
PubMed1985

Lethality of intestinal tissue extracts from Eimeria-infected chickens.

Evidence exists that lethal activity is associated with extracts from Eimeria tenella-infected caecal tissues. We examined tissues of the small intestine infected with other species of Eimeria for a similar lethal activity. Extracts were prepared from intestinal tissues of chickens infected with either E. acervulina, E. brunetti, E. maxima, E. mitis or E. necatrix. Appropriate control extracts from tissues of non-infected chickens were also prepared. Intravenous injection of the extracts into healthy chickens showed that lethality of infected tissue extracts was higher than control extracts and was independent of the site of infection within the intestine. Comparison of the lethality of caecal extracts from chickens infected with embryo-adapted or parent lines of E. tenella showed that lethality was independent of the recognized pathogenic characteristics of the lines. Also, it was demonstrated that lethal activity is associated with extracts of tissues containing massive numbers of arrested and non-arrested sporozoites and early trophozoites.

Animals↗
PubMed1984

[Lipoamide dehydrogenase, citrate synthase and beta-hydroxyacyl-CoA-dehydrogenase in skeletal muscle. I. Determination of activity in tissue extracts].

It is to be expected that changes in the subcellular distribution of the mitochondrial enzymes lipoamide dehydrogenase (LIPDH), citrate synthase (CS) and beta-hydroxyacyl-CoA-dehydrogenase (HADH) in the muscle tissue give information on the type and the extent of damage of mitochondria during storage and treatment of meats; such changes may be also used as basis of methods for the differentiation between fresh and frozen/thawed meat. Standard methods for the determination of the activities of LIPDH, CS, and HADH in tissue extract and muscle press juice are described. The influence of enzyme concentration, pH and temperature on the enzyme activities in muscle extract was investigated. Furthermore the error in the enzyme analyses by the standard methods was determined.

Acyl-CoA Dehydrogenase↗
PubMed2001

Methods of tissue extraction in advanced laparoscopy.

Advances in laparoscopic surgery have revolved not only around new methods of tissue excision and repair, but also around specimen extraction. At the center of these recent advances is the need to be cost-effective and safe. Two particularly important advances in laparoscopic tissue extraction are use of electronic power morcellators, to cut up and remove large masses, and specimen bags to remove the extracted tissue safely. Although morcellators offer tremendous time savings, and can actually reduce the risk of hernia formation because fascia need not be torn or stretched, the use of power morcellators is not without concern. The sharp rotating blade can damage normal organs. Furthermore, one must watch closely so that morcellated specimens are not inadvertently misplaced. Specimen bags are also important, both in malignant and in benign conditions. Ultimately, the problem is spillage of tissue, whether it be cancer cells or products of conception. Although some specimen bags are equipped with self-opening devices, the largest bags must be opened manually.

Biopsy↗
PubMed1993

Immunoreactive hepatocyte growth factor is present in tissue extracts from human breast cancer but not in conditioned medium of human breast cancer cell lines.

Hepatocyte growth factor (HGF) is a novel mitogen for mature hepatocytes. In the present study, we have measured immunoreactive (ir)-HGF concentration in tumor extracts of 82 primary human breast cancers using an enzyme-linked immunosorbent assay (ELISA). Ir-HGF was detectable in all tissue extracts, the concentration ranging from 1.4 to 306.5 ng/100 mg protein (median value: 11.2 ng/100 mg protein). Correlation analyses between ir-HGF concentration and clinicopathological factors showed that the ir-HGF level was significantly higher in tumors with sizes of more than 5.0 cm compared with those less than 5.0 cm. In contrast, no detectable amount of ir-HGF was secreted into culture medium of two breast cancer cell lines, MCF-7 and ZR-75-1, suggesting that the cancer cell itself has no ability to produce ir-HGF.

Breast Neoplasms↗
PubMed1985

Evaluation of microtitre plates for the routine assay of oestrogen and progesterone receptors in tissue extracts.

A time- and material-saving assay of oestrogen and progesterone receptors in tissue extracts using enzyme-linked immunosorbent assay quality microtitre plates and microtitre-compatible equipment is described. The results from simultaneous determinations in microtitre plates and in conventional tubes show the former to yield higher receptor values. Furthermore, the results of a large series of measurements in microtitre plates compare favourably with previously obtained results from assays in tubes.

Breast Neoplasms↗
PubMed2006

KCC isoforms in a human lens epithelial cell line (B3) and lens tissue extracts.

We recently reported potassium-chloride cotransporter activity in human lens epithelial B3 (HLE-B3) cells. The purpose of the present study was to demonstrate in these cells as well as in human lens tissue the potassium-chloride cotransport (KCC) isoforms by reverse transcriptase-polymerase chain reaction (RT-PCR), Western blotting and immunofluorescence microscopy. Of the four KCC genes known to encode the respective proteins and their spliced variants, RT-PCR with both rat and human primers revealed the predicted cDNA fragments of KCC1, KCC3a, KCC3b, and KCC4 but not KCC2 in both HLE-B3 cells and in human lens tissue extracts from cataractous patients. Polyclonal rabbit (rb) anti-rat (rt) and anti-human (hm) antibodies against rtKCC1 and hmKCC3, respectively, and a commercially available rb-anti-mouse (ms) KCC4 antibody were used. Rb anti-rtKCC1-ECL3 [against epitopes within the large extracellular loop 3 (ECL3)] revealed a 150kDa band in HLE-B3 cells consistent with the known molecular weight of KCC1. Rb anti-hmKCC3-ECL3 yielded three bands of 150, 122 and 105kDa, evidence for the presence of KCC3a, KCC3b and possibly KCC3c isoforms. The 122 and 112kDa bands were also demonstrated by rb anti-hmKCC3-CTD [the C-terminal domain (CTD)]. Rb anti-msKCC4 antibody only showed a 100kDa band in HLE-B3 cells. In the human lens tissues, a 115kDa protein was detected with rb anti-rtKCC1-ECL3 and a 100kDa band with rb anti-msKCC4, however, no bands with rb anti-hmKCC3-ECL3 or rb anti-hmKCC3-CTD. Fluorescence microscopy revealed immunocytochemical cytoplasmic and membrane labeling of HLE-B3 cells with anti-KCC1, -KCC3 (laser confocal microscopy) and -KCC4 antibodies and a Cy3-tagged secondary antibody. Hence HLE-B3 cells expressed proteins of the KCC1, KCC3a, b, and KCC4 isoforms, whereas surgically removed cataractous lens tissue expressed only those of KCC1 and KCC4.

Blotting, Western↗
PubMed1985

A rapid tissue extraction method for determining liver copper content by atomic absorption spectroscopy.

A simple method using tissue extraction of Copper (Cu) in 8M nitric acid for 45 minutes at 100 degrees C, followed by sample dilution and Cu quantitation by flame atomic absorption spectroscopy is described. The detection limit of the method is 16 micromol Cu/kg liver. The mean recovery of added Cu was 100.4%. This method for liver copper analysis correlated well with a method using nitric/perchloric acid digestion and atomic absorption spectroscopy (r2 = .986) and is faster, less expensive and less hazardous.

Journal Article↗
PubMed1985

New enzymatic changes of L-cystathionine catalyzed by bovine tissue extracts.

L-Cystathionine was used as substrate for enzyme systems prepared by heating bovine tissue extracts in the presence of pyruvate at 60 degrees C for 10 min. Analysis of the products indicated that the systems converted L-cystathionine into the cyclic ketimine form which was detected by its spectral properties and by chromatography on the amino acid analyzer. Alanine, alpha-aminobutyrate and cystine were also produced. Pyruvate and alpha-ketobutyrate enhance the production of the ketimine by liver, kidney and heart extracts, and are necessary for the brain extracts: alpha-Ketoglutarate is much less effective and its presence favors the production of homocystine by all the extracts. Homocystine was found in the brain incubates when any of the ketoacids assayed were added. The overall reaction is explained by the action of heat stable cystathionine gamma-lyase and beta-synthase which produce alpha-ketobutyrate and pyruvate used for the transamination of the remaining cystathionine to the monoketoacid. This last compound cyclizes spontaneously into the ketimine form thus avoiding the removal of the second amino group. This represents a new nontransulfurative path leading to the production of a seven membered etherocyclic product whose biochemical implications are yet unexplored.

Amino Acids↗
PubMed1992

DNA extraction by sonication: a comparison of fresh, frozen, and paraffin-embedded tissues extracted for use in polymerase chain reaction assays.

DNA extraction from fixed tissues can be the most laborious and complex step in amplifying DNA by the polymerase chain reaction (PCR). We have previously reported a rapid and efficient method for extracting DNA by the use of sonication and glass beads. We have extended our experiences with this technique using fresh, frozen, and formalin-fixed paraffin-embedded tissues with and without the use of glass beads and report their results. Multiple tissue types were obtained at autopsy or as part of a surgical specimen. DNA was extracted from identical tissue when the sample was fresh, frozen, or formalin-fixed paraffin-embedded. Our results indicate that in most instances the sonication technique, which takes only 30 min from start to finish, can rapidly extract fresh, frozen, or formalin-fixed paraffin-embedded tissue and is superior to other rapid extraction techniques in terms of quality and quantity of DNA. It is much more rapid than those techniques that use long digestion periods. This technique will be of great value to those investigators extracting DNA for polymerase chain reaction assays.

DNA↗
PubMed1986

Automated high-performance liquid chromatographic method with column switching for the determination of neurotransmitters and related compounds, ascorbic acid and uric acid in tissue extracts.

An automated high-performance liquid chromatographic method with electrochemical and fluorimetric detection and on-line data evaluation is described for the simultaneous measurement of indoleaminergic and catecholaminergic neurotransmitters, some of their metabolites and precursors and ascorbic and uric acids. Deproteinized tissue extracts from the central nervous system or peripheral organs are injected without prior purification (recovery greater than 90%). A switching system enables the compounds to be passed as necessary through one, two or three reversed-phase columns, which are then eluted simultaneously (analysis time 25 min). Fifty samples per day can be analysed with a precision of 95% for neurotransmitters and about 90% for ascorbic and uric acids.

Animals↗
PubMed2004

Development of an immunofluorometric assay and quantification of human kallikrein 7 in tissue extracts and biological fluids.

BACKGROUND: Human kallikrein 7 (hK7), also known as human stratum corneum chymotryptic enzyme, is a chymotrypsin-like serine protease first identified in human skin extracts and predicted to be a secreted protease. The aim of this study was to develop a sensitive and specific immunoassay for hK7 and to examine the distribution of hK7 in tissue extracts and biological fluids. METHODS: Recombinant hK7 was produced in human embryonic kidney cells (HEK293T) and purified by a three-step column chromatographic procedure. The purified hK7 was injected into mice for antibody generation. A sandwich-type immunoassay was developed with the anti-hK7 monoclonal antibodies. RESULTS: The assay had imprecision (CV) <10% through the dynamic range of 0.2-20 microg/L and had no detectable cross-reactivity from other members in the human kallikrein gene family. Highest concentrations were found in skin, esophagus, and kidney. hK7 was also found in amniotic fluid, ascites from ovarian cancer patients, breast milk, cerebrospinal fluid, saliva, seminal plasma, serum, sweat, synovial fluid, and urine. CONCLUSIONS: This study describes the first ELISA-type immunoassay for hK7 protein quantification. hK7 is found many human tissues and in various biological fluids.

Adult↗
PubMed1984

Enhancing activity of rat tissue extracts for induction of lambda prophage by L-azaserine.

We studied the effect of rat tissue extracts on induction of lambda prophage in Escherichia coli (lambda) by L-azaserine. Hepatic and pancreatic extracts, primarily the cytosolic fraction, markedly increased the rate of induction. Hepatic extracts from lipotrope-deficient rats were somewhat more active than extracts from normal rats. The enhancing activity in normal rat hepatic cytosol was partially characterized. It reduced by about one-half the dose of azaserine required for a given purpose. The enhancement was increased by preincubating the bacterial cells with cytosol; cells retained the effect after cytosol was removed. Enhancing activity was inhibited strongly by the amino acids phenylalanine, tryptophan, and tyrosine; to lesser extents by leucine, methionine, and serine; and not at all by proline or glutamine. It was eliminated by dialysis of the cytosol and reduced by omission of nicotinamide adenine dinucleotide phosphate (NADP) from the reaction mixture. Heating the cytosol to 60 degrees C or 80 degrees C or varying the pH of the reaction mixture from 6 to 8 had no significant effect. Treating the cytosol with trypsin appeared to release an inhibitor of the activity. Glutathione, cysteine, and beta-mercaptoethanol also enhanced lambda induction by azaserine, but the cytosolic activity was not affected by the thiol-inactivating compound diethylmaleate (DEM). The results suggest that factors in cytosol interact with bacterial cells to facilitate transport of azaserine into the cells, primarily through the aromatic amino acid transport system. A small molecule, not a free thiol compound, appears to be involved. It may serve to establish reducing conditions protective for azaserine, the probable mechanism of action of sulfhydryl compounds.

Animals↗
PubMed1991

Induction of experimental murine granuloma formation against Schistosoma japonicum eggs produced by in vitro ova deposition, in vitro tissue extraction, or lyophilization.

Fresh eggs, laid by Schistosoma japonicum adult worms in vitro or recovered from tissue after trypsin digestion, as well as non-viable lyophilized eggs were injected into mice via the cecal vein. The freshly laid eggs induced a marked, maximal reaction at 2 weeks after implantation. This reaction was indistinguishable from that seen early in naturally infected mice. Lyophilized eggs induced granulomas only one-tenth the size of those formed around freshly laid eggs. Maximal granulomatous reactivity was not seen until the 4th week after implantation and the intensity of cellular reactivity and associated histopathological change was much lower than that observed with freshly laid eggs. Reactions against live, tissue-extracted eggs were quantitatively and temporally intermediate between those observed against laid and those surrounding lyophilized eggs. The results suggests that these differences in granuloma formation are due to variable quantities of locally produced cytotoxic materials and/or antigen that stimulate immune reactions of different intensities.

Animals↗