Envenomation by the scorpion Centruroides exilicauda (C sculpturatus): severe and unusual manifestations.
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Animals↗
Biomedical subjects
R A Berg
Publications and source records attributed to R A Berg.
Activities of a cancellous bone-cell-stimulating substance.
A bone-cell-stimulating substance (BCSS) that initiates appositional bone formation in intact rats was examined for its effects on DNA and collagen synthesis in tibial and calvarial organ cultures of 17-day-old embryonic chicks. BCSS stimulated collagen synthesis in both types of bone. BCSS stimulated DNA synthesis in tibiae but inhibited synthesis in calvaria from the same chicks. Insulinlike growth factor, transforming growth factor-beta, platelet-derived growth factor, and fetal bovine serum also affected DNA synthesis differently in calvaria and tibiae. BCSS was able to modify some of the effects of these growth factors on DNA.
Animals↗
Retinoids affect collagen synthesis through inhibition of ascorbate-induced lipid peroxidation in cultured human dermal fibroblasts.
Ascorbate has been shown to stimulate collagen synthesis in cultured human dermal fibroblasts by increasing transcription of the collagen genes. In the present studies, ascorbate stimulates lipid peroxidation at concentrations similar to those necessary to affect collagen synthesis. Molecules which inhibit lipid peroxidation, such as propyl gallate, cobalt chloride, and alpha-naphthol, also inhibit collagen synthesis, suggesting a correlation between the two phenomena. Retinoic acid and some synthetic retinoids have previously been shown to inhibit collagen synthesis in cultured human dermal fibroblasts. In our studies two different retinoids, at similar concentrations, inhibit both ascorbate-stimulated lipid peroxidation and collagen synthesis. Since high concentrations of retinoids were required, the ability of retinoids to inhibit the oxidant effect of ascorbate, and not their receptor-mediated activity, may be responsible for their effect on collagen synthesis.
Antioxidants↗
Protein disulphide isomerase from human peripheral blood neutrophils.
Protein disulphide isomerase (PDI) is a 56 kDa resident polypeptide of the endoplasmic reticulum of many cell types. We evaluated the ability of human peripheral blood polymorphonuclear neutrophils (PMN) to synthesize both mRNA and proteins. Using in vitro [35S]-methionine labeling of purified PMN, followed by immunoprecipitation of cell lysates with immobilized polyclonal and monoclonal antibodies and analysis by gel electrophoresis, PMN were shown to synthesize many proteins, including actin. In contrast, incorporation of [35S]-methionine into PDI was not detected. Purification of total RNA from PMN and analysis by Northern blots demonstrated the presence in PMN of PDI-RNA. Western immunoblot evaluations of total PMN protein display an immunoreactive-PDI of 56 kDa. Indirect immunofluorescence studies suggest an abundance of immunoreactive-PDI throughout PMN. We therefore conclude that PDI is synthesized in precursor cells of the bone marrow. Phorbol 12-myristate 13-acetate, a reagent known to affect the degranulation of specific granules, causes the release of immunoreactive-PDI into a post-centrifugation supernatant. PDI, a ubiquitous endoplasmic reticulum resident protein, is shown here to be associated with specific granules in a cell type which has lost its intracellular membrane network during terminal differentiation.
Animals↗
Mechanical stress and cellular metabolism in living soft tissue composites.
A soft tissue substitute composed of a three-dimensional collagen lattice infiltrated with living fibroblasts was modelled as a two-phase composite material. The phases consisted of an extracellular matrix containing fibres and fluid and cellular inclusions. A mathematical model that had been previously derived for calculating the elastic stress and energy distribution around spherical inclusions and voids in a three-dimensional elastic medium was applied to a biological composite containing cells in an elastic medium. The model predicted an increase in stress inside the cells in the direction of applied load and a decrease in the direction perpendicular to the applied load. The model also predicted increased interfacial stress at the cell-matrix boundary in the direction of applied load with maximum and minimum values at different points on the cell periphery. It was hypothesized that the interfacial stress around the cell inclusions might play a function in signal transduction. To test this hypothesis, an average unidirectional stress was applied to the boundary of a 1 cm X 2 cm X 3 mm lattice containing fibroblasts. When cells were cultured for 7 d and placed under stress of 15 g, cell growth was increased 1.7 fold, protein synthesis decreased 48% and intracellular cAMP was increased 3.7 fold in 24 h. The changes were a function of externally applied stress. Cells cultured for 14 d and placed under stress displayed similar results. The results supported the concept that soft tissue extracellular matrix depicted as an elastic medium can affect cell growth and development.(ABSTRACT TRUNCATED AT 250 WORDS)
Animals↗
Pulsus paradoxus in the diagnosis and management of pneumopericardium in an infant.
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Blood Pressure↗
Occult carbon monoxide poisoning in an infant.
The diagnosis of occult carbon monoxide poisoning in children is a difficult problem and may be frequently overlooked. An infant with recurrent episodes of unrecognized CO exposure and subsequent neurologic deterioration is presented. A clue to the diagnosis in this case was the finding of basal ganglia abnormalities on computed tomography. Clinical findings, etiology, and treatment of CO poisoning are reviewed. We suggest that CO toxicity should be suspected in any child with acute or recurrent neurologic deterioration.
Basal Ganglia Diseases↗
Collagen and elastin metabolism in hypertensive pulmonary arteries of rats.
We evaluated the processes controlling the accumulation of collagen and elastin in main pulmonary arteries of rats during an episode of hypoxic pulmonary hypertension. Explant cultures of main pulmonary arteries were incubated with [3H]proline to measure collagen and protein synthesis and percent collagen synthesis. Elastin synthesis was measured by [14C]valine incorporation into insoluble elastin. Relative collagen synthesis increased twofold (from 1.1 +/- 0.2 x 10(3) to 2.0 +/- 1.0 x 10(3) disintegrations per minute [14C]hydroxyproline/vessel/hr/mg protein), relative collagen synthesis doubled (from 2% to 4-5% of total protein synthesis), and elastin synthesis increased ninefold (from 0.4 +/- 0.2 x 10(4) to 3.6 +/- 0.6 x 10(4) dpm [14C]valine/vessel/hr/mg protein) in early hypertension. The level of pro alpha l(I) collagen RNA paralleled the relative collagen synthetic rate during the study period. Within 7 days of recovery from hypoxia, collagen and elastin contents were normal. We conclude that collagen and elastin in main pulmonary arteries are synthesized rapidly during an episode of hypoxic pulmonary hypertension and that collagen and elastin are rapidly removed from the hypertensive vessel during normoxic recovery.
Actins↗
Activation of neutrophils by factors released from alveolar macrophages stimulated with collagen-like polypeptides.
Oxidant-induced injury is associated with breakdown of interstitial collagen and the accumulation of inflammatory cells in the lungs. In previous studies, we demonstrated that phagocyte accumulation is mediated, in part, by chemotactic factors generated from damaged collagen. To determine if alveolar macrophages also mediate the migration of polymorphonuclear leukocytes (PMN) into the lungs, we examined the release of chemotactic factors from alveolar macrophages treated with native or synthetic collagenous peptides. These included fragments of bovine collagen digested with bacterial collagenase (CG) or cyanogen bromide (CB) as well as small molecular weight synthetic polypeptides containing proline (Pro), glycine (Gly), and hydroxyproline (Hyp), the major amino acids that comprise collagen. We found a dose- and time-related generation of PMN chemotactic activity by collagen peptide-treated macrophages. The maximum activity was released 72 h after pretreatment of macrophages for 1 to 3 h with 0.1 to 1 microM CG-, CB-, or (Pro-Pro-Gly)5-peptides. The native peptides derived from CG-digested collagen were more active than synthetic peptides containing Pro and Gly. Neither trypsin digests of bovine serum albumin nor synthetic peptides containing Hyp stimulated chemotactic factor release from macrophages. The alveolar macrophage-derived chemotactic factor was found to lose activity when dialyzed and after heat or trypsin treatment. The release of PMN-activating factors by collagen peptide-treated macrophages was also examined. Alveolar macrophage-conditioned medium was found to stimulate PMN production of reactive oxygen intermediates as well as elastase and gelatinase.(ABSTRACT TRUNCATED AT 250 WORDS)
Amino Acid Sequence↗
Activation of neutrophil collagenase by cathepsin G.
Collagenase is secreted from neutrophils as a latent or proenzyme. In an effort to understand the mechanism of collagenase activation in inflammation, human peripheral neutrophils (PMNs) were isolated and incubated with the tumor promotor, phorbol myristate acetate (PMA), which induces the neutrophils to degranulate and secrete proteinases. Neutrophil media were then treated with various activators or inhibitors of collagenase and other proteinases, and the collagenase activity was measured. A serine proteinase secreted from neutrophils, cathepsin G, was found to activate latent collagenase, but it was also found to require activation itself. Both hypochlorous acid (HOCl) and oxidized glutathione (GSSG) were tested for their collagenase-activating ability and were found to be successful only in the presence of active cathepsin G. A specific cathepsin G inhibitor (0.5 mM Z-Gly-Leu-Phe-CH2Cl) prevented the activation of latent collagenase by HOCl. To confirm these results, purified neutrophil cathepsin G was incubated with a neutrophil proteinase mixture which contained latent collagenase. The collagenase was shown to be activated upon incubation with purified cathepsin G. These results indicate that cathepsin G is a key mediator in neutrophil collagenase activation.
Animals↗
Cathepsin G degrades denatured collagen.
Neutrophils are known to contain several metalloproteinases that can damage collagen, a major structural component of the extracellular matrix. Here a neutrophil serine proteinase secreted from activated neutrophils was shown to cleave denatured collagen (gelatin). This serine proteinase was not inhibited by synthetic inhibitors of elastase (elastatinal or Me-O-suc-Ala-Pro-Val-CH2Cl). However, a synthetic inhibitor of cathepsin G (Z-Gly-Leu-Phe-CH2Cl) was able to inhibit the serine proteinase having gelatinolytic activity, indicating that cathepsin G, a major serine proteinase, from neutrophils is responsible for cleaving gelatin. Purified cathepsin G was also shown to degrade gelatin. In further experiments, oxidized glutathione was able to enhance the gelatinolytic activity of cathepsin G. These results show that cathepsin G is capable of cleaving denatured collagen, and its activity is enhanced or stabilized in the presence of glutathione. The data support the concept that cathepsin G released from neutrophils could play a major role in degrading collagen during inflammation and may in part account for the degradation of extracellular matrix during inflammation.
Animals↗
Hypochlorous acid (HOCl) activation of neutrophil collagenase requires cathepsin G.
In an effort to understand the mechanism of collagenase activation in inflammation, human peripheral neutrophils were isolated and incubated with the tumor promoter, phorbol myristate acetate (PMA), which induces the neutrophils to degranulate and secrete proteinases. Neutrophil media were then treated with HOCl with or without various proteinase inhibitors then collagenase activity was measured. Added HOCl was able to activate latent collagenase. However, a serine proteinase, cathepsin G, was found to be necessary for collagenase activation to occur by HOCl. The results indicate that cathepsin G is a key mediator in neutrophil collagenase activation and that HOCl under certain conditions leads to the activation of cathepsin G or the stimulation of cathepsin G's ability to activate neutrophil collagenase.
Animals↗
Effect of proteoglycans on type I collagen fibre formation.
Collagen fibrillogenesis is a multistep process involving assembly of molecules into fibrils and bundles of fibrils. The exact role of proteoglycans in collagen fibrillogenesis is unclear. The purpose of these studies is to study the effect of proteoglycans on collagen fibrillogenesis in vitro. Results of these studies suggest that the proteoglycans dermatan sulphate and chondroitin sulphate do not change the final turbidity and hence the diameter of fibrils formed during the early stages of fibrillogenesis. This suggests that proteoglycans may not influence the early phases of collagen assembly, such as nucleation. However, proteoglycans added during the final stages of collagen fibre formation in vitro cause changes in ultimate tensile strength. In the presence of the high-molecular-weight proteoglycan, the ultimate tensile strength is increased by a factor of 1.5 above that of the control, whilst in the presence of low-molecular weight chondroitin sulphate proteoglycan the tensile strength is significantly decreased. It is concluded that proteoglycans influence the later stages of fibre formation. The presence of high-molecular-weight chondroitin sulphate proteoglycan leads to efficient stress transfer between collagen fibrils, altering the ultimate tensile strength. The results of these studies will be useful in optimizing the design of collagen tendon-ligament prostheses.
Animals↗
Protein disulphide isomerase, a multifunctional endoplasmic reticulum protein.
Protein disulphide isomerase (E.C. 5.3.4.1) has been purified, cloned, and sequenced from a variety of vertebrate tissues. The enzyme and its isoforms have been assigned a role in four functional activities: (1) hydroxylation of proline residues in procollagen; (2) disulphide bond oxidation, isomerization, and reduction; (3) the major non-nuclear binding protein of the thyroid hormone 3,3',5-triiodo-L-thyronine; and (4) a component of oligosaccharide transferase. The concentration of the enzyme has been shown to be positively correlated with an endoplasmic reticulum network which is active in secreting disulphide-bonded polypeptides. The enzyme is directed into the endoplasmic reticulum by virtue of a 19 residue N-terminal signal peptide; a four amino acid C-terminal KDEL sequence prevents the enzyme from being secreted. Careful inspection of the sequence data of the isoforms from human tissues reveals a 97% similarity; whereas, analyses of the data from chick tissues reveals only a 80% level of similarity. Chromosomal localizations using human cDNA probes against different human isoforms have assigned the gene(s) to opposite ends of the long arm of chromosome 17. The compiled data suggest the presence of a family of related polypeptides, all of which reside within the lumen of the endoplasmic reticulum.
Amino Acid Sequence↗
Prolyl 4-hydroxylase: molecular cloning and the primary structure of the alpha subunit from chicken embryo.
Prolyl 4-hydroxylase (EC 1.14.11.2) is a key enzyme required for the posttranslational hydroxylation of proline residues in collagen. The enzyme is a tetramer composed of two pairs of nonidentical subunits (alpha 2 beta 2). The beta subunit is protein disulfide-isomerase, a ubiquitous enzyme found in the endoplasmic reticulum of many cell types. We report here the amino acid sequence of the alpha subunit. One cDNA clone (alpha 1) was isolated from a chicken embryo cDNA expression library in lambda gt11 by screening with anti-alpha-subunit polyclonal immunoglobulins. This alpha 1 cDNA contains an open reading frame of 1401 base pairs. A comparison of the translation of the nucleotide sequence with protein sequences obtained from the purified chicken alpha-subunit polypeptide verified that alpha 1 cDNA encoded the alpha subunit. Polymerase chain reactions were used to extend the sequence of alpha 1 cDNA toward the 5' end of alpha-subunit mRNA. The mature alpha subunit is composed of 516 amino acids with a calculated molecular mass of 59,373 Da. The compiled amino acid sequence contains two potential glycosylation sites, an observation that agrees with a previous demonstration that the alpha subunit contains two N-linked oligosaccharide chains. Blot hybridization analysis of total chicken embryo RNA detected an mRNA of 3.5 kilobases, a size that closely resembles the size of the cloned cDNA. Since the expression of the alpha subunit is confined to cell types that synthesize and secrete collagens, the regulation of the synthesis of the alpha subunit may play a central role in determining the expression of prolyl 4-hydroxylase activity.
Amino Acid Sequence↗
Medical management of fungal suppurative thrombosis of great central veins in a child.
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Amphotericin B↗
Effects of compound 48/80 and exogenous histamine on wound healing in mice.
Compound 48/80 has previously been shown to improve wound healing in rats, presumably through stimulation of histidine decarboxylase activity and mobilization of histamine from mast cells. In the present study, C57Bl/6 mice were wounded by dorsal skin incision followed by treatment with compound 48/80, exogenous histamine, or the combination of 48/80 plus histamine. Skin-breaking strength was significantly increased over saline-injected controls by the combined treatment with 48/80 and histamine. Neither 48/80 or histamine alone had any influence on wound healing. Histamine content of skin at the wound site was significantly reduced by 48/80 treatment, but was unaffected by 48/80 plus histamine or histamine given alone. In contrast, stomach and leg muscle histamine levels were significantly increased beyond those of unwounded, wounded saline- or 48/80-injected mice. These results were also confirmed in CD mice, and are in contrast to findings in rats in which treatment with 48/80 alone significantly improved wound healing of similarly injured animals.
Animals↗
Wound healing using a collagen matrix: effect of DC electrical stimulation.
Rapid fibroblast ingrowth and collagen deposition occurs in a reconstituted type I collagen matrix that is implanted on full-thickness excised animal dermal wounds. The purpose of this study is to evaluate the effects of direct current stimulation on dermal fibroblast ingrowth using carbon fiber electrodes incorporated into a collagen sponge matrix. Preliminary results suggest that fibroblast ingrowth and collagen fiber alignment are increased in collagen sponges stimulated with direct currents between 20 and 100 microA. Maximum fibroblast ingrowth into the collagen sponge is observed near the cathode at a current of 100 microA. These results suggest that electrical stimulation combined with a collagen matrix may be a method to enhance the healing of chronic dermal wounds.
Animals↗