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Biomedical subjects

J H Reese

Publications and source records attributed to J H Reese.

33 records · Page 2Linked to original sources
PubMed1987

Identification and purification of a kidney membrane protein which specifically binds the amino-terminal domain of native parathyroid hormone.

Nitrocellulose blots of bovine kidney membrane proteins were prepared from denaturing polyacrylamide gels. Strips of the blots were incubated with parathyroid hormone (PTH), washed, and then incubated with antisera against the hormone. Exposure to horseradish peroxidase-linked second antibody led to staining of a 51-kDa protein. No staining was observed in blots not incubated with PTH. Fragments 35-84 and 19-84 of PTH reacted strongly with the antisera, but did not lead to staining of the 51-kDa protein on the blots. Staining was visible, but greatly reduced, when fragment 9-84 was used. Oxidation of the native hormone at positions 8 and 18 led to reductions in staining of the band which were quantitatively similar to the reductions in biological activity induced by such oxidations. These properties suggested that the 51-kDa protein recognizes the amino-terminal portions of PTH, which is the segment of the molecule required for its biological activities. Several micrograms of the 51-kDa protein were purified to homogeneity by selective extraction from the membranes with detergent and by elution from multiple two-dimensional gels. The purified protein retained its PTH-dependent staining and specificity. This protein may be a PTH receptor or a fragment of a PTH receptor from kidney.

Animals↗
PubMed1987

Monoclonal antibodies with defined specificities for Torpedo nicotinic acetylcholine receptor cross-react with Drosophila neural tissue.

A panel of monoclonal antibodies with known specificity for the well-characterized nicotinic acetylcholine receptor from the electroplax of Torpedo californica, many of which cross-react with the mammalian muscle acetylcholine receptor, were examined for cross-reactivity in the fly, Drosophila melanogaster. Monoclonal antibodies with specificities for different epitopes on the transmembrane receptor complex from Torpedo cross-react with different regional subsets of neural tissue in Drosophila. Axonal tracts, neuropil, mechano-sensory bristle elements and photoreceptors, each are detected by separate monoclonal antibody classes corresponding to different epitope domains. A preliminary characterization of an antigenic determinant in Drosophila heads recognized by one of the cross-reacting monoclonal antibodies is presented. Monoclonal antibodies such as these may be useful in identifying molecules of homologous structure or function, possibly including a neuronal acetylcholine receptor.

Animals↗
PubMed1987

Phyllodes type of atypical prostatic hyperplasia: a report of 3 new cases.

We report 3 new cases of phyllodes type of atypical prostatic hyperplasia. This lesion is characterized by epithelial and stromal proliferation. Stromal changes are the most characteristic finding in phyllodes type of atypical prostatic hyperplasia, which show atypical cells with enlarged, hyperchromatic sarcomatoid nuclei. Mitotic figures are not present. Although the histological appearance may mimic that of cystosarcoma phyllodes of the breast, this pattern is present only focally or not at all in phyllodes type of atypical prostatic hyperplasia. On computerized tomographic imaging phyllodes type of atypical prostatic hyperplasia has a distinct appearance. These patients can be expected to have a benign clinical course and distant metastases have not been reported. Treatment is by surgical excision as in benign prostatic hyperplasia.

Adult↗
PubMed1986

Cribriform adenocarcinoma of the prostate.

In 46 radical prostatectomy specimens, carcinoma volume and Gleason histologic grade were compared among 21 cancers containing Grade 3 cribriform areas and 25 noncribriform carcinomas. All cribriform cancers but only 44% of the noncribriform cancers were larger than 1.7 ml. Grade 4-5 areas were present in 86% of cribriform cancers and 36% of noncribriform cancers. All 17 cancers larger than 4.0 ml contained Grade 4-5 areas, and 15 of these showed cribriform areas. By contrast, noncribriform Grade 3 areas were found in 45 of 46 carcinomas. In most cases, cribriform carcinoma was predominantly intraductal; cell masses either followed normal duct contours or showed a basal cell layer by routine microscopic study or by immunohistochemical staining for basal cell-specific keratin. Seven cases showed primary origin of cribriform carcinoma within ducts by evolution from intraductal dysplasia, a presumptive premalignant lesion. It was proposed that cribriform carcinoma is equivalent to intraductal Grade 4 cancer, and that many Grade 4 prostate cancers may evolve directly from an identifiable intraductal precursor.

Adenocarcinoma↗
PubMed1986

Alpha-bungarotoxin binding to a high molecular weight component from lower vertebrate brain identified on dodecyl sulfate protein-blots.

The binding of [125I]iodo-alpha-bungarotoxin [( 125]alpha-BuTX) to the dissociated alpha-subunit of Torpedo acetylcholine receptor (AChR) can be readily demonstrated in a modified 'protein-blot' analysis utilizing electrophoretically transferred, dissociated subunits immobilized onto positively charged nylon membranes which are then incubated directly with [125I]alpha-BuTX. We report here the use of the protein-blotting technique to detect the alpha-BuTX binding site present in the central nervous system of lower vertebrates and to characterize some of the physicochemical properties of the toxin binding site. High molecular weight (Mr greater than or equal to 200,000 and greater than or equal to 120,000) alpha-BuTX-binding components can be readily demonstrated in avian and fish brain extracts upon protein-blotting with [125I]alpha-BuTX following lithium dodecyl sulfate PAGE. Neither extensive reduction with dithiothreitol nor prior reduction followed by alkylation with iodoacetamide alter the mobility of the CNS-derived BuTX-binding sites. In contrast to our findings with Torpedo AChR or muscle AChR derived from a number of different species, no binding is observed in the molecular weight range of the alpha-subunit (Mr = 40,000) nor is any binding at any molecular weight observed in similar fractions prepared from adult, mammalian (rat, guinea pig) brain using this technique. These results demonstrate the existence in lower vertebrate brain of a BuTX binding site comparable in size to the AChR oligomeric complex of electric organ and muscle. They also suggest, however, striking structural differences between muscle AChR and the central neuronal BuTX-binding complex as well as a considerable difference between the neuronal BuTX-binding sites derived from lower and higher vertebrate brain.

Animals↗
PubMed1986

Differential distribution of pepsinogen II between the zones of the human prostate and the seminal vesicle.

Pepsinogen II (PG II) is a gastric proenzyme which has previously been found in both human seminal fluid and the prostate gland. However, no regional distribution of PG II has been noted within the prostate nor has it been found in the seminal vesicle. Bouins-fixed sections of central zone, peripheral zone and seminal vesicle, taken from 10 prostates removed at radical prostatectomy or cystectomy, were exposed to antibody against PG II and stained using the A-B-C immunoperoxidase technique. Formalin-fixed tissue from autopsy prostates of four men in the third decade, and six cases with BPH nodules, were also examined for PG II activity. In nine of 10 seminal vesicles, and seven of 10 central zone samples, more than 50 per cent of the cells stained positive for PG II. By contrast, in nine of 10 peripheral zone samples staining was present in five per cent or less of the epithelial cells. Similarly, PG II activity in the four autopsy prostates occurred almost entirely within the central zone and ended abruptly at the boundary between the peripheral and central zones. BPH nodules contained no PG II activity. These findings provide the first evidence that the central and peripheral zones may serve different biological functions. Embryologically it is currently thought that the prostate is of endodermal origin and the seminal vesicle of mesodermal origin. The presence of large amounts of PG II in both the seminal vesicle and central zone lends support to the hypothesis of a common mesodermal origin for these two structures.

Aged↗
PubMed1984

Stimulation of acetylcholine release from guinea-pig ileal synaptosomes by cyclic nucleotides and forskolin.

Various agents which are known to affect intracellular levels of cAMP have been assessed for their ability to induce the release of [3H]acetylcholine ([3H]ACH) from a synaptosomal preparation derived from the guinea-pig ileum myenteric plexus. 8-Bromo-cAMP increased the release of [3H]ACh above basal levels. While 8-bromo-cGMP also increased the release, this nucleotide was far less potent than 8-bromo-cAMP. Comparison of the release caused by the cyclic nucleotides to the release induced by the nicotinic agonist dimethylphenylpiperazinium (DMPP) suggested that there is some relationship, as yet undefined, between the 8-bromo-cAMP-induced and the DMPP-induced release, while no relationship was evident between the release induced by 8-bromo-cGMP and that caused by DMPP. The 8-bromo-cAMP-induced release was Ca2+-dependent. Neither adenosine, clonidine, nor oxotremorine (all of which modulate the nicotinically-induced release) affected the 8-bromo-cAMP-induced release. The phosphodiesterase inhibitor 3-isobutyl-1-methylxanthine stimulated the release of [3H]ACh as did the adenylate cyclase activator forskolin. The forskolin-induced release was not affected by adenosine, clonidine or oxotremorine. The ability of the modulators to block the nicotinically-induced release but not the release caused by the cyclic nucleotides indicates that the modulation of release evoked by nicotinic activity does not occur at a step involving protein phosphorylation.

1-Methyl-3-isobutylxanthine↗
PubMed1984

Opiate binding to subcellular fractions from guinea pig ileum.

Binding of 3H-etorphine and 3H-D-Ala2-D-Leu5-enkephalin to opiate receptors in synaptosomal and microsomal fractions prepared from guinea pig ileum homogenates has been studied. It is found that the dissociation constants for etorphine from all fractions are the same. The binding capacity for etorphine for the purified synaptosomal fraction is greater than for other fractions by a factor of 5. For the enkephalin derivative binding to the microsomal fraction the dissociation constant is greater than for etorphine while the binding capacity is a factor of 3 lower. These results are in contrast to the case for binding to central nervous system subcellular fractions.

Animals↗
PubMed1983

Activation of fluoride-stimulated adenylate cyclase by phospholipase A2 in the caudate nucleus of the rat brain.

Phospholipase A2 (PLA2) increases adenylate cyclase (AC) activity in the rat caudate nucleus in a dose-dependent manner. After maximal stimulation by fluoride, PLA2 treatment further increases AC activity 2.4 fold. Adenylate cyclase activity is maximal after 45% hydrolysis of the phospholipids. Of the products of PLA2 treatment only lysophosphatidylcholine (LPC) produces such an increase in AC activity. In contrast to PLA2 treatment, LPC solubilizes the enzyme, decreases the Km value for ATP, and requires much larger amounts of LPC than that produced by lipase treatment. After maximal stimulation with fluoride and PLA2, removal of most of the LPC does not reduce the activity of adenylate cyclase. These findings suggest that removal of membrane lipid rather than generation of LPC is responsible for the activation of brain adenylate cyclase by phospholipase A2.

Adenylyl Cyclases↗
PubMed1982

Modulation of the release of acetylcholine from ileal synaptosomes by adenosine and adenosine 5'-triphosphate.

Low concentrations of either adenosine or adenosine 5'-triphosphate (ATP) inhibited the nicotinically induced release of [3H]acetylcholine from synaptosomes derived from the guinea-pig ileum myenteric plexus. Adenosine and ATP were equipotent in their ability to inhibit the release and the inhibition was reversible by theophylline in both cases. The data suggest that ATP may have acted after initial hydrolysis to adenosine and that the receptor involved may be the P1 or similar R site receptor. High concentrations of ATP caused marked increases in the release of [3H]acetylcholine. This release was neither temperature- nor calcium-dependent. Because the concentrations required were similar, however, to those which have been reported to cause ATP-induced contractions in intact preparations, further studies of the phenomenon were carried out. Lactate dehydrogenase was not released with the [3H]acetylcholine, suggesting that indiscriminate lysis of the membranes had not occurred. The release was not affected by theophylline, indomethacin, tetrodotoxin or adenosine. The ATP analog, adenylyl (beta, gamma-methylene)-diphosphonate did not cause the increase in release, therefore phosphorylation may be required for the effect. The mechanism of increased release remains to be defined, but the data suggest that it is unlikely that a P2 receptor is involved.

Acetylcholine↗
PubMed1977

Binding of apomorphine to neural membranes.

The intrinsic fluorescence of apomorphine has been used to measure its binding to neural membranes. A large number of relatively weak binding sites are concentrated in myelin and synaptic membrane fractions. Butyrophenones have the highest affinities for these sites--KD = 43 micrometer for haloperidol--while dopamine and dopamine releasers and reuptake blockers, as well as a variety of other alkaloids, have much lower affinities. The sites are hydrophobic and undergo a phase transition to a highly fluid state near 26 degrees C. Calcium is a noncompetitive inhibitor of apomorphine binding. Some of the actions of neuroleptic drugs may result from binding to these hydrophobic membrane sites in vivo, blocking conduction in small catecholamine axons.

Animals↗
PubMedDate not supplied

Post-translational modification of proteins by 15-carbon and 20-carbon isoprenoids in three mammalian cell lines.

A number of cellular proteins, including p21ras, lamin B, and the G-protein gamma subunits, undergo post-translational modification by 15-carbon farnesyl or 20-carbon geranylgeranyl isoprenoid moieties derived from pyrophosphate intermediates of the cholesterol biosynthetic pathway. In this study, isoprenylated proteins in three mammalian cell lines (Hela cells, Rat-6 fibroblasts and COS cells) were radiolabeled with an isoprenoid precursor, [3H]mevalonate, and resolved by SDS gel electrophoresis. Groups of proteins with different molecular masses were eluted from the gels and the chain-lengths of the radiolabeled isoprenyl groups, released from the proteins by Raney-nickel-catalyzed desulfurization, were established by gel permeation chromatography. 15-Carbon and 20-carbon isoprenyl groups were found in separate classes of proteins within each cell line. With the exception of p21ras, which incorporated a 15-carbon group when expressed in COS cells, the proteins in the region of the 21-28 kDa ras-related GTP binding proteins contained mostly 20-carbon isoprenyl chains. In contrast, proteins belonging to the 66-72 kDa nuclear lamin family, as well as unidentified proteins with molecular masses of 41-46 kDa and 53-55 kDa, contained predominantly 15-carbon isoprenyl chains. The chain-lengths of the isoprenoids associated with particular classes of proteins did not vary from one cell line to another, suggesting that the nature of the isoprenoid modification (farnesyl versus geranylgeranyl) is determined by intrinsic structural features of the proteins, rather than the cell type in which the proteins are expressed.

Animals↗
PubMedDate not supplied

A study of the interaction between dirithromycin and astemizole in healthy adults.

The effect of a standard regimen of dirithromycin, a macrolide antibiotic, on the single-dose pharmacokinetics of the H (1) receptor blocker astemizole was evaluated in a sample of 18 healthy young adults (nine males and nine females). The study was conducted in a two-way cross-over fashion after the subjects had been randomly given either dirithromycin (two 250 mg tablets) or placebo (two tablets) every morning for 10 days. On the morning of the fourth dose of either dirithromycin or placebo each subject ingested a single 30-mg oral dose (three 10-mg tablets) of astemizole. The disposition kinetics of both astemizole and its major metabolite, N-desmethylastemizole, were characterized after measuring the concentrations of both analytes in the serum fraction of serial blood samples collected for 14 days after the astemizole dose. In addition, corrected QT (QT(c) ) intervals were estimated from electrocardiogram rhythm strips that were run 24 hours prior to the astemizole dose, 12 hours after the astemizole dose, and after the last treatment (dirithromycin or placebo) dose in both study periods. Pharmacokinetic parameters that were measured for both astemizole and N-desmethylastemizole during each treatment were: C(max), t(max), AUC (0-infinity), CL(oral), half-life, and volume of distribution (V). None of the parameters for N-desmethylastemizole was different when comparing data by ANOVA from the dirithromycin treatment period with that of the placebo treatment period. On the other hand, during dirithromycin treatment astemizole CL(oral) was 34% slower, volume of distribution was 24% larger, and half-life was 84% longer. Generally, all QT ( c ) intervals did not appear to be affected by dirithromycin treatment. The changes in astemizole kinetics could not be attributed to its N-demethylation since the dispositional kinetics of N-desmethylastemizole were unaffected by dirithromycin. Therefore, it is difficult to ascertain the clinical significance of the changes in astemizole kinetics. Since there were no significant differences for mean QT(c) intervals and no effect of dirithromycin treatment on N-desmethylastemizole kinetics, it is unlikely that a standard regimen of dirithromycin would place a patient taking astemizole at an increased risk of torsade de pointes or related ventricular arrhythmias.

Adolescent↗