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Biomedical subjects

J McConnell

Publications and source records attributed to J McConnell.

89 records · Page 5Linked to original sources

Storage and long-term use of human biopsy tissue in histochemical studies.

Since a common problem in histochemical methods is the need for fresh-frozen tissue, techniques have been developed in this laboratory for the long-term storage and use of frozen tissue. Cryostat sections of tissue stored for as long as 4 years at -70 degrees C have been utilized to localize biogenic amines and acetylcholinesterase (AChE). The quality of the tissue and the histochemical demonstration of peripheral nerve fibers in the stored specimens is comparable to that seen in fresh-frozen tissue. Procedures are outlined for storing the tissue prior to cutting sections, for freezing and mounting the tissue for cutting, and for long-term storage of tissue blocks. These methods have been used successfully for organs of the mammalian urogenital system of several different mammals, including man, and for the brain, spinal cord, pituitary, adrenal, thyroid and submandibular glands of the rat. The methodology should be applicable to most organs and systems.

Acetylcholinesterase↗

Biosynthesis of Trypanosoma brucei variant surface glycoproteins - analysis of carbohydrate heterogeneity and timing of post-translational modifications.

The variant surface glycoproteins from two cloned populations of Trypanosoma brucei brucei which were known to migrate as multiple bands on SDS gels have been studied. The heterogeneity present was located in those oligosaccharide side chains the addition of which is tunicamycin-sensitive. The time required for the trypanosome to synthesize and express a variant surface glycoprotein molecule in vitro was found, from pulse-chase and limited trypsinisation experiments, to be approximately 40 min. In the light of these data, pulse-chase experiments on the two antigens known to have heterogeneity in their oligosaccharide side chains demonstrated that the heterogeneity probably arose by two different mechanisms. Pulse-chase experiments on three different clones of trypanosomes have also been used to investigate the timing of cleavage of the carboxyl-terminal extension, known to be encoded on variant surface glycoprotein mRNA. Similar pulse-chase experiments followed by immunoprecipitation using affinity purified antiserum have been used to investigate the addition of the cross-reacting determinant. The timing of both these events has been discussed in relation to the time necessary for the synthesis and expression of the variant surface glycoprotein on the surface of the trypanosome.

Animals↗

The vasculature of the human penis: a reexamination of the morphological basis for the polster theory of erection.

The morphology of human penile blood vessels has been studied to characterize columns of smooth muscle found in the intima. Although previous workers (Kiss, 1921; Conti, 1952) termed these intimal pads or polsters and concluded that they were actively responsible for the production of erection, more recent work has questioned these conclusions (Benson et al., 1980; Newman and Tchertkoff, 1980). Serial sections were taken throughout the length of the penis from 11 cadavers and stained with various histopathological stains. Numerous polster-like structures were found, but all could be identified as either branch points in vessels, intimal cushions located at branch points, or arteriosclerotic lesions at various stages of development. None of these structures seem capable of functioning in the manner attributed to them by Conti (1952) and others. Furthermore, comparison of the structures found in the present study to illustrations of polsters in Conti's paper suggest that these are identical. The present study refutes the polster theory of erection and leaves open to further question the mechanisms involved in the control of blood flow into and within the penis.

Adult↗

The biosynthesis of Trypanosoma brucei variant surface glycoproteins--in vitro processing of signal peptide and glycosylation using heterologous rough endoplasmic reticulum vesicles.

The ability of stripped microsomal dog pancreas vesicles to process precursors of the variant glycoprotein of Trypanosoma brucei has been investigated. The vesicles were shown to be capable of cleaving a peptide precursor variant surface glycoprotein, identified as the N-terminal signal peptide in the case of one variant. The vesicles were also shown to add carbohydrate to precursor variant surface glycoprotein. This carbohydrate, which was sensitive to the action of endo-H was located within a C-terminal CNBr fragment in one variant. However, the vesicles were not capable of adding the oligosaccharide carrying the variant surface glycoprotein cross-reacting determinant.

Animals↗

Biosynthesis of Trypanosoma brucei variant surface glycoprotein. I. Synthesis, size, and processing of an N-terminal signal peptide.

An N-terminal signal peptide has been identified on a precursor of the variant surface glycoprotein (VSG) of Trypanosoma brucei, synthesised both in vitro and in vivo. An ordered cyanogen bromide (CNBr) cleavage map was constructed for both mature VSG and for precursor VSG, and this allowed a size estimate of the signal peptide to be made by comparison of the N-terminal CNBr peptides produced from each. The signal peptide contains 30-40 amino acids. Analysis of the precursor VSGs produced when nascent polypeptide chains on T. brucei polysomes were completed in the absence of reinitiation showed that the signal peptide is removed before translation is complete and that the same translational start signal is utilised in the trypanosome and in the rabbit reticulocyte protein synthesis system.

Amino Acid Sequence↗

Neuromorphology and neuropharmacology of the human penis: an in vitro study.

The neuromorphology and neuropharmacology of the human penis are only briefly described in literature. The present study was undertaken to define the adrenergic and cholinergic neuromorphology of the human corpus cavernosum (CC) and corpus spongiosum and to evaluate the in vitro response of the CC to pharmacologic stimulation. Human penile tissue was obtained from six transsexual patients undergoing penectomy. For morphologic study, the tissue was processed for (a) hematoxylin and eosin staining; (b) smooth muscle staining; (c) acetylcholinesterase localization; (d) glyoxylic acid histofluorescence; (e) electron microscopy; and (f) electron microscopy after glutaraldehyde dichromate fixation. In addition, strips of CC were placed in in vitro muscle chambers and tension changes recorded isometrically after stimulation with norepinephrine (NE) and acetylcholine. The CC contains abundant smooth muscle, numerous glyoxylic acidfluorescent (catecholaminergic) fibers and varicosities, and a scant distribution of acetylcholinesterase-positive fibers. Fewer of all these elements were present in the corpus spongiosum. No "polsters" were observed in the CC. Although glutaraldehyde-fixed controls exhibited no typical adrenergic vesicles (small, dense core, measuring 400-600 A in diameter), some small, strongly electron-dense vesicles were found in glutaraldehyde dichromate-fixed tissue and were thought to contain NE. A variety of other vesicles were also encountered. The addition of NE to the in vitro muscle chambers caused a dose-related contraction, which was blocked by pretreatment with phentolamine in all CC strips tested. Acetylcholine in high concentration produced minimal contraction in 2 of 24 strips. Our morphologic and pharmacologic data suggest that the sympathetic nervous system may affect erection by acting not only on the penile vasculature but also by direct action on the smooth muscle of the CC itself.

Acetylcholine↗

Autonomic innervation of the mammalian penis: a histochemical and physiological study.

The distribution of adrenergic and cholinergic nerves was studied in penile tissue from rat, rabbit, cat, monkey and man. Glyoxylic acid histofluorescence and acetylcholinesterase-positive fibers were found in all tissues examined, but histofluorescent, presumably adrenergic fibers predominated. In general, except in the rabbit, more nerve fibers of both types were found in the corpus cavernosum (CC) than in the corpus spongiosum/penile urethra (CS). The rabbit penis showed slightly more adrenergic fibers in the CC than in the CS and more cholinergic nerves in the CS than in the CC. The CC of the monkey demonstrated an alpha receptor mediated contractile response to norepinephrine (NE) stimulation and no response to acetylcholine (ACh) in an in vitro muscle bath.

Acetylcholinesterase↗

Estrogenic activity of the insecticide chlordecone (Kepone) and interaction with uterine estrogen receptors.

The chlorinated insecticide chlordecone (Kepone) interacts with the estrogen receptor system in the rat uterus in vitro and in vivo. It competes with estradiol for binding to the cytoplasmic receptor in vitro and also induces nuclear accumulation of estrogen receptor sites in uteri in vitro. When injected into immature rats, chlordecone translocates estrogen receptor sites to the uterine nucleus, increases uterine weight, and stimulates the synthesis of the progesterone receptor, an estrogen receptor-mediated process. Its slow onset of action but prolonged duration of interaction with estrogen receptor and stimulation of uterine weight gain and progesterone receptor synthesis indicates that, although it has an affinity for receptor only 0.01-0.04% that of estradiol, its considerable estrogenic activity may likely be derived from its long half-life and bioaccumulative character.

Animals↗

Dual effects of concealed A-V nodal conduction in man.

An interpolated premature ventricular contraction (PVC) may produce either complete block of the next sinus impulse or depression of A-V nodal conduction with a prolonged A-H interval. When a PVC results in partial depression of a A-V nodal conduction, the effect on subsequent premature atrial stimuli is unknown. The authors have recently observed three patients in which the effect of a premature ventricular stimulus with interpolation on the functional refractory period of the A-V node could be measured. In case one an interpolated PVC sufficient to prolong the A-H interval from 80 to 120 msec was followed by programmed premature atrial stimuli which resulted in no additional A-V nodal delay, and the apparent functional refractory period of the A-V node was reduced from 420 to 330 msec when compared with the atrial extrastimulus technique. In case two a programmed ventricular extrastimulus prolonged the A-H interval in the following sinus beat from 120 to 240 msec; atrial extrastimuli then resulted in only minimal increments in A-V nodal delay and the apparent functional refractory period of the A-V node was reduced from 590 msec. A ventricular extrastimulus in case three increased the resting A-H interval from 60 to 115 msec; conduction of atrial extrastimuli then resulted in a reduction in the functional refractory period of the A-V node from 465 to 400 msec. In each case an interpolated premature ventricular stimulus produced (1) depression of A-V nodal conduction in the ensuing sinus beat A1 and (2) relative facilitation of A-V nodal conduction of a subsequent premature atrial stimulus (A2). The functional refractory period of the A-V node was reduced when compared with the atrial extrastimulus technique alone.

Atrioventricular Node↗

Autonomic innervation of the urogenital system: adrenergic and cholinergic elements.

The major organs of the male urogenital (UG) system have been examined in various mammals, including man, using light and electron microscopic (EM) histochemical methods. For the light microscopic study, the urinary bladder, the vas deferens and the penis (corpora cavernosa and corpus spongiosum) were studied in the rat, cat, dog, monkey and man using a glyoxylic acid (GA) method modified for peripheral adrenergic nerve fibers, and a thiocholine method for acetylcholinesterase (AChE). Fine structural analysis was done on the vasa of rat, cat, monkey and man, and on the bladder and penis of cat, dog, monkey and man. Tissue was fixed in glutaraldehyde (GMO) as a control or in glutaraldehyde-dichromate (GDC) for the specific localization of norepinephrine (NE). All organs studied demonstrated numerous adrenergic nerve fibers throughout the muscular layers, in the connective tissue, and in the adventitia of most blood vessels. These fibers had a brilliant fluorescence when visualized with the GA method, and demonstrated many varicosities with small (400-600 A) and/or large (800-1200 A) granular vesicles in both control and GDC-fixed tissue examined with the EM. Evaluation of the vesicles with the analytical electron microscope (AEM) verified that those in the GDC-fixed tissue were chrome-positive, and, therefore, NE-containing. In the vas and penis, acetylcholinesterase(AChE)-positive nerve fibers were encountered less frequently at the light microscopic level than adrenergic fibers, and few typical cholinergic varicosities were seen in these organs with the EM. In the bladder, cholinergic nerves were seen with about the same frequency as adrenergic fibers in both light microscopic and EM preparations. Also observed frequently in each of the viscera were varicosities with large to very large (800-2000 A) granular vesicles of the kind presently hypothesized to be peptidergic or purinergic. Few varicosities of the type considered sensory, with large (800-1200 A) clear vesicles and numerous mitochondria, were observed in this tissue. Evidence from this study suggests that mammalian UG organs are innervated extensively by adrenergic nerves, and, excepting the bladder, have a limited cholinergic innervation; in the bladder, numerous fibers of each type can be found. In addition, another type of nerve fiber, perhaps peptidergic or purinergic, is found in large numbers in each of the organs studied and thus may represent a significant effector of autonomic regulation.

Acetylcholinesterase↗

Cobalamin deficiency associated with erythroblastic anemia and methylmalonic aciduria in a border collie.

Anemia due to cobalamin deficiency is a rare genetic disorder that has been recognized in dogs only recently. This report concerns a 14-month-old border collie that presented for chronic, nonregenerative anemia. Cytological examination of a peripheral blood smear showed the presence of erythroblasts. Serum cobalamin levels were below reference ranges reported for clinically normal dogs. A methylmalonic aciduria was found on urinalysis. These signs are consistent with the anemia in Imerslund-Graesbeck syndrome reported in humans. Anemia due to cobalamin deficiency responds to parenteral vitamin B12 therapy, and affected animals have a good prognosis for recovery.

Animals↗