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Biomedical subjects

D J Fletcher

Publications and source records attributed to D J Fletcher.

At least 55 records · Page 3Linked to original sources
PubMed1983

Catfish somatostatin is unique to piscine tissues.

It has been previously shown that catfish islets contain two distinct forms of somatostatin: somatostatin-14 and the predominant form, catfish somatostatin, which is a 22-residue peptide structurally related to somatostatin-14. Using antisera against this catfish somatostatin and somatostatin-14, other tissues of the catfish and pancreatic tissue of various animals were examined for the presence of these two peptides. Catfish intestine also contained large amounts of catfish somatostatin in comparison to that of somatostatin-14, but the predominant form in catfish brain tissues was somatostatin-14. Relatively small quantities of catfish somatostatin were found in extracts of anglerfish islet, but none were detected in pancreatic tissues of frog, chicken, or rat. Somatostatin-14 was found in relatively large amounts in these other pancreata. These results suggest that catfish somatostatin is found only in piscine tissues and that there may be a differential expression of the two somatostatin genes in those tissues.

Animals↗
PubMed1983

Hormone release by islet B cell-enriched and A and D cell-enriched populations prepared by flow cytometry.

Dispersed pancreatic islet cells were analyzed for their low forward angle light scatter using flow cytometry. The cells produced a distinct light scatter pattern which appeared to be a function of cell size and not cell granularity. RIA of hormone content of cells collected from different regions of the pattern revealed that glucagon- and somatostatin-containing cells were concentrated in regions of lower scatter intensity and that insulin-containing cells were more numerous in regions of higher intensity. Relative to the original cell suspension, these preparations were enriched 3-fold in glucagon and somatostatin content and 6-fold in insulin content. The function of intact islets, unsorted dispersed cells, and sorted dispersed cells was examined before and after 4 days of culture. Before culture, all of the dispersed cell populations had elevated basal secretion compared with intact islets and did not respond to stimulatory concentrations of glucose, arginine, or 3-isobutyl-1-methylxanthine. After culture for 4 days, basal secretion fell, and responsiveness returned. In both the A/D cell-enriched and the B cell-enriched cultured populations, the percentage of single cells was approximately 95%. The insulin release patterns from these populations were similar to those from intact islets and unsorted dispersed cells. Glucagon release from all of the dispersed cell populations far exceeded that from intact islets. This study suggests that the structural organization of islets influences A cell function, but a clear influence upon B cell function has not been demonstrated.

1-Methyl-3-isobutylxanthine↗
PubMed1982

Hair analysis. Proven and problematic applications.

Hair analysis has important uses in screening for metal intoxication and exposure to environmental pollutants, as well as applications in forensic medicine. In addition, hair analysis has been hailed as the new "in" tool for diagnosis of nutritional deficiencies and chronic diseases, but data to support these applications are fragmentary at best.

Cystic Fibrosis↗
PubMed1981

Characterization of proinsulin- and proglucagon-converting activities in isolated islet secretory granules.

The conversion of proglucagon and proinsulin by secretory granules isolated from both prelabeled and unlabeled anglerfish islets was investigated. Either granules isolated from tissue labeled with [3H]tryptophan and [14C]isoleucine or [35S]cysteine, or lysed granules from unlabeled tissue to which exogenously labeled prohormones had been added were incubated under various conditions. Acetic acid extracts of these granule preparations were analyzed for prohormone and hormone content by gel filtration. Both prelabeled and lysed, unlabeled secretory granules converted radiolabeled precursor peptides (Mr 8,000-15,000) to labeled insulin and glucagon. The accuracy of the cleavage process was established by demonstrating comigration of products obtained from in vitro cleavage with insulin and glucagon extracted from intact islets using electrophoresis and high-pressure liquid chromatography (HPLC). The pH optimum for granule-mediated conversion was found to be in the range of pH 4.5-5.5. Conversion of both proglucagon and proinsulin by secretory granules was significantly inhibited in the presence of antipain, leupeptin, p-chloromercuribenzoate (PCMB) or dithiodipyridine (DDP) but not chloroquine, diisopropyl fluorophosphate, EDTA, p-nitrophenyl guanidinobenzoate, soybean trypsin inhibitor, or N-p-tosyl-L-lysine chloromethyl ketone HCl. The inhibitory action of PCMB and DDP was reversed in the presence of dithiothreitol. Both membranous and soluble components of the secretory granules possessed significant converting activity. HPLC and electrophoretic analysis of cleavage products demonstrated that the converting activities of the membranous and soluble components were indistinguishable. The amount of inhibition of proinsulin and proglucagon conversion caused by 600 micrograms/ml porcine proinsulin was significantly lower than that caused by the same concentration of unlabeled anglerfish precursor peptides. These results indicate that the proinsulin and proglucagon converting enzyme(s) in the anglerfish pancreatic islet is a unique intracellular thiol proteinase(s) that may be granule membrane-associated and may require the presence of prohormone sequences in addition to the dibasic residues at cleavage sites for substrate recognition and/or binding.

Animals↗
PubMed1978

Somatostatin biosynthesis occurs in pancreatic islets.

No information is at present available on the mode of SRIF biosynthesis. Since anglerfish pancreatic islet tissue is comprised of approximately 30% D cells, we have examined this tissue for SRIF synthesis . The following known differences in amino acid composition of islet peptides were used advantageously in this study: anglerfish proinsulin: Trp-0, Ile-2, Cys-6; anglerfish glucagon: Trp-1, Ile-0, Cys-0; mammalian SRIF: Trp-1, Ile-0, Cys-2. After incubating islet tissue with [3H]tryptophan and [14C]isoleucine or [35S]cystine for various time periods, proteins were extracted in 2 M acetic acid and desalted by Bio-Gel P-2 gel filtration. P-2 void volume proteins were then subjected to P-10 gel filtration and isolated by polyacrylamide gel electrophoresis (PAGE) at alkaline pH. The predominant amount of the immumoreactive SRIF in the extracts appeared in a peak eluting just before the salt volume on P-10 filtration and migrated slowly toward the cathode during PAGE. The behavior of synthetic SRIF was identical. The anglerfish SRIF immunoreactive peptide could be labeled with Trp and Cys but not Ile during incubations longer than 1 h. The Trp- and Cys-labeled peptide could be bound on columns to which the immunoglobulin fraction of antisera to SRIF had been complexed. Cycloheximide inhibited isotope incorporation into all islet proteins. These results indicate that islet SRIF is synthesized in situ. Moreover, the immunological activity, size, and charge characteristics of anglerfish islet SRIF appear to be similar to those of mammalian hypothalamic SRIF. When islets were subjected to short pulse incubations with labeled Trp and Cys, only peptides eluting in the 7,000-13,000 dalton portion of the filtration eluate became labeled. No appreciable isotope incorporation into SRIF was observed. However, when pulse incubations were followed by incubation in the presence of cycloheximide or excess unlabeled amino acids in isotope-free medium (chase), the incorporation of Trp and Cys into SRIF increased with the length of chase, suggesting the participation of a larger precursor in SRIF synthesis.

Animals↗
PubMed1976

Immunohistochemical localization of somatostatin, insulin and glucagon in the principal islets of the anglerfish (Lophius americanus) and the channel catfish (Ictalurus punctata) (1) (2).

Somatostatin, insulin and glucagon were localized in the principal islets of the anglerfish (Lophius americanus) and the channel catfish (Ictalurus punctata) by means of the unlabeled antibody-peroxidase-antiperoxidase immunocytochemical method. Both species showed a similar ratio of positive cells 9:6:4 (insulin:somatostatin:glucagon), but the interrelations of the three cell types differed between species. The large number of somatostatin-positive cells may be indicative of an important role for this hormone in teleost physiology. The principal islets appear to be a good source of tissue for further work on somatostatin.

Animals↗
PubMed1975

P-32 uptake the letic algae.

A study of the Flat Creek Embayment of Lake Sidney Lanier near Gainesville, Georgia revealed three genera of algae, Chlorococcum, Fragillaria and Nostoc, to be prominent in this eutrophic region of the lake. The algae was grown in phosphate-rich media and subsequently labelled with P-32. All species incorporated luxury amounts of phosphorus as determined by the uptake of P-32. The results indicate that the P-32 uptake is proportional to the surface-per-volume ratio. The higher surface-per-volume ratio resulted in greater uptake of P-32.

Eukaryota↗