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Biomedical subjects

D Chang

Publications and source records attributed to D Chang.

At least 217 records · Page 12Linked to original sources
PubMed1989

Purification and biochemical characterization of CP4 (SP-D), a collagenous surfactant-associated protein.

CP4 is a collagenous glycoprotein (43 kDa, reduced) synthesized by rat type II pulmonary epithelial cells in primary culture (Persson et al., 1988). In order to better characterize this protein, CP4 was isolated from rat bronchoalveolar lavage and EDTA extracts of lung surfactant by adsorption to barium sulfate and elution with sodium citrate followed by reverse-phase HPLC. Amino acid analysis of purified CP4 demonstrated 4-hydroxyproline (Hyp), hydroxylysine (Hyl), and acid-labile components coeluting with Hyl glycosides. In addition, gas-phase amino-terminal microsequencing of two CP4 CNBr peptides demonstrated nonoverlapping collagenous sequences comprised of nine and six Gly-X-Y triplets, containing a total of four residues of Hyp and two of Hyl. There was less than 50% sequence homology of these peptides with the cDNA-derived sequence of the collagenous domain of rat SP-A. Two-dimensional IEF/SDS-PAGE resolved the protein into a charge train of basic isoforms (pI approximately 6-8), similar to those of newly synthesized CP4 and the class D surfactant proteins (Phelps & Taeusch, 1985). Gel filtration of nondenatured CP4 on 4% agarose showed a high apparent molecular mass complex comprised of disulfide-bonded trimers of the 43-kDa subunits. Antibodies to purified lavage CP4 showed specific binding to newly synthesized and surfactant-associated CP4. We propose that CP4 be designated "surfactant protein D" (SP-D) in accordance with an accepted nomenclature for surfactant-associated proteins.

Amino Acid Sequence↗
PubMed1989

Isolation and characterization of bovine pancreastatin.

Bovine pancreastatin, a 47 amino acid residue peptide, was isolated from the pancreas and the pituitary gland using a chemical method which detects its C-terminal glycine amide structure. The complete amino acid sequence of the pancreatic peptide is 74% homologous to that of porcine pancreastatin and is identical to bovine chromogranin A-(248-294), as deduced from its cDNA sequence. The sequence of the first 28 amino-terminal residues of the pituitary peptide was determined to be identical to the corresponding sequence of the pancreatic peptide. Since the pituitary peptide also contains the C-terminal glycine amide, it is therefore likely to be identical in structure to the pancreatic peptide. Thus, we conclude that bovine chromogranin A is the precursor of bovine pancreastatin. Synthetic bovine pancreastatin inhibited pancreatic exocrine secretion in a similar manner to porcine pancreastatin.

Animals↗
PubMed1989

Regulation of collagen production by medial smooth muscle cells in hypoxic pulmonary hypertension.

Pulmonary hypertension is associated with abnormal connective tissue deposition in the media of pulmonary arteries. Lobar arteries from calves maintained for up to 15 days at simulated high altitude showed a 35% increase in collagen and a greater than 40% increase in crosslinked elastin per microgram protein. Labeling of artery tissue with [14C]proline revealed a nearly twofold increase in relative collagen synthesis. There was increased incorporation into Types I, III, IV, and V collagen with an increase in the proportion of newly synthesized Type IV collagen. Quantitation of collagen mRNA by slot-blot assay demonstrated increased levels of Types I and IV collagen message. In addition, medial smooth muscle cells isolated from the hypertensive calves demonstrated a nearly twofold increase in relative collagen synthesis, a twofold increase in the accumulation of newly synthesized collagen per microgram DNA, and increased levels of Types I and IV collagen mRNA. Exposure of pulmonary artery smooth muscle cells, adventitial cells, and fetal calf ligament fibroblasts to conditioned calf serum harvested from cultures of medial cells from hypertensive animals increased their levels of collagen as well as elastin mRNA. These studies suggest that the increased production of collagen in hypertensive arteries is mediated at a pre-translational level by soluble factor(s) generated by medial smooth muscle cells.

Animals↗
PubMed1989

Effects of endothelin on porcine coronary arterial strips.

Endothelin, a novel endothelium derived 21-residue vasoconstrictor peptide synthesized by Peninsula Laboratories, provoked a concentration-dependent contraction of porcine coronary arterial strips. EC50 value for endothelin was 14 +/- SD 4 nmol/L (n = 6), and significantly lower than the values for 5-hydroxytryptamine (5-HT, 0.28 +/- 0.07 mumol/L, n = 6) and 15-methyl-prostaglandin F2 alpha (15-methyl-PGF2 alpha, 4 +/- 3 mumol/L, n = 7). The maximal increase in tension caused by endothelin was 5.4 +/- 1.1 g, being much greater than that induced by 5-HT (3.7 +/- 0.8 g, P less than 0.05) and 15-methyl-PGF2 alpha (3.7 +/- 0.6 g, P less than 0.01). The changes in tension provoked by endothelin (2-20 nmol/L) were attenuated significantly after pretreated with tetrodotoxin (TTX, 30 mumol/L, P less than 0.05 or 0.01). The results suggest that endothelin is one of the most potent vasoconstrictive agents, and its action is partially related to voltage-sensitive Na+ channel in the cell membrane.

Amino Acid Sequence↗
PubMed1989

Host genetics as a cause of overdispersion of parasites among hosts: how general a phenomenon?

In the white-footed mouse, Peromyscus leucopus, the tapeworm Hymenolepis citelli occurs at low (2-3%) prevalence in the field. We found that mature infections (i.e., with egg production) developed in up to 100% of hosts. In the laboratory, a majority of hosts lost their infection by 28 days postintubation. In wild mice infected in the laboratory and returned to the field, infections were more prolonged, with half of the mice still infected at 100 days postintubation. A majority of previously infected hosts resisted challenge infection. Our introduction of laboratory-infected mice into a natural population of hosts appeared to cause infections among previously uninfected mice, leading to an increase in the prevalence of tapeworm infection among mice not intubated. Although genetically based expulsion of tapeworms before maturity is important in causing low prevalence in a similar host-parasite system, such resistance cannot explain low prevalence in the present system. It appears that both heterogeneous distribution and rarity of intermediate hosts as well as short parasite lifespan contribute to low prevalence and overdispersion. Host-parasite dynamics of 2 very similar systems appear to differ markedly.

Animals↗
PubMed1988

CP4: a pneumocyte-derived collagenous surfactant-associated protein. Evidence for heterogeneity of collagenous surfactant proteins.

Type II pneumocytes secrete pulmonary surfactant and are known to synthesize SP-35, a collagenous surfactant-associated protein. Freshly isolated type II cells also synthesize other bacterial collagenase-sensitive and hydroxyproline-containing proteins, including a glycoprotein designated CP4. CP4 was isolated from rat pneumocyte culture medium by immune precipitation with polyclonal antibodies to rat surfactant proteins or by DEAE chromatography and reverse-phase or gel permeation HPLC. CP4 did not cross-react with polyclonal antibodies to SP-35 and was completely resolved from SP-35 by SDS-PAGE (Mr 43K reduced) or isoelectric focusing. Unlike SP-35, which consists of acidic isoforms assembled as disulfide-bonded dimers and multimers, CP4 was secreted as basic isoforms assembled as disulfide-bonded trimers. Differences in primary structure were demonstrated by CNBr and V8 protease peptide mapping. The secretion of both proteins was inhibited by 2,2'-dipyridyl, an inhibitor of posttranslational prolyl and lysyl hydroxylation and collagen triple helix formation. CP4 was isolated from EDTA extracts of rat surfactant. These studies provide evidence for the heterogeneity of pneumocyte-derived collagenous surfactant-associated proteins.

Animals↗
PubMed1988

Calcium: a program in BASIC for calculating the composition of solutions with specified free concentrations of calcium, magnesium and other divalent cations.

A BASIC program is presented which facilitates the formulation of biologically relevant chemical solutions containing specified free concentrations of as many as three divalent metal cations (Ca2+, Mg2+ and the choice of a third divalent cation) at any pH in the presence of as many as three ligands (EGTA, ATP and GTP). The program uses the law of mass action and the absolute stability (association) constants found in the literature to calculate the total concentration of divalent metal cation needed to achieve a desired free concentration. The user enters the pH, the concentrations of the ligands used and the desired free concentrations of the divalent cations. This program was developed for use in a wide range of biological applications, particularly the rapid design of solutions which mimic certain aspects of intracellular fluid.

Adenosine Triphosphate↗
PubMed1988

A sensitive radioimmunoassay for somatomedin-C/insulin-like growth-factor I based on synthetic insulin-like growth factor 57-70.

We have developed a region-specific radioimmunoassay for somatomedin-C/insulin-like growth factor-I (IGF-I) directed against IGF-I 57-70. This sequence includes part of the A region and all of the carboxyterminal D region. In addition to being highly specific, this immunoassay is sensitive to as little as 1 ng/ml of IGF-I, and has half-maximum displacement by 10 ng/ml of IGF-I. Comparison to other existent immunoassays for IGF-I shows correlation coefficients of greater than 0.95. This immunoassay has the advantage of being directed against a known region of the IGF-I molecule which is on the exterior of the 3-dimensional structure, and can use a readily available synthetic peptide as both radioligand and standard.

Antibody Specificity↗
PubMed1988

Digitonin-permeabilized colonic cell layers. Demonstration of calcium-activated basolateral K+ and Cl- conductances.

Sheets of isolated turtle colon were exposed to digitonin on the mucosal side to chemically remove the apical membrane as a permeability barrier. Increases in the mucosal uptake of 86Rb, [3H]mannitol, and 45Ca-EGTA, and the appearance of the cytosolic marker enzyme lactate dehydrogenase in the mucosal bath confirmed the permeabilizing effect of the detergent. Basolateral K+ and Cl- currents were generated by imposing transmural ion gradients, and cytosolic free Ca2+ was manipulated by means of a Ca2+-EGTA buffer system in the mucosal bathing solution. Raising the cytosolic free Ca2+ concentration from the nanomolar to the micromolar range activated basolateral conductances for K+ and Cl-. Differences in ion selectivity, blocker specificity, calcium activation kinetics, and divalent cation activation selectivity indicated that the Ca2+-induced increases in the K+ and Cl- conductances were due to separate populations of channels. The results are consistent with the notion that the apical membranes of turtle colon epithelial cells can be functionally removed under conditions that preserve some of the conductive properties of the basolateral membrane, specifically Ca2+-activated conductive pathways for K+ and Cl-. This permeabilized preparation should offer a means for the identification of macroscopic currents that are due to presumed Ca2+-activated channels, and may also provide a model system for the functional reconstitution of channel regulatory mechanisms.

Animals↗
PubMed1988

Solitary non-parasitic liver cyst treated by cyst-gastrostomy.

A 66 year old female presented with a symptomatic solitary non-parasitic liver cyst. This was treated by a cyst-gastrostomy following which the patient has remained well without complications for 12 years. Given an appropriate anatomical relationship of cyst and stomach, this would appear to be a safe and effective operation.

Cysts↗