Inhibition of human pancreatic elastase II activity on human aortic elastin by human α2-macroglobulin

Summary Human 2-macroglobulin-human pancreatic elastase II binding were investigated using a homologous substrate, human aortic elastin, in order to test the enzymatic activity. We demonstrated that two moles of 2-M are required to inhibit one mole of HPE_II when the enzyme is added to a mixture of elastin and 2-M. In addition, when the elastase-2-M complex is prepared under some circumstances, it exhibits an elastinolytic activity.     

Newly synthesized elastin is associated with neoplastic epithelial cells in human mammary carcinoma

Summary Indirect immunofluorescence with a purified antiserum to human foetal elastin has identified newly synthesized elastin on the membranes of neoplastic epithelial cells in human mammary carcinoma.This research was supported by a project grant from the Medical Research Council, London.     

Magnesium enhances human pancreatic elastase digestion of 125I-labeled elastin

The effect of some divalent cations, especially Mg++, on elastinolysis by porcine or human pancreatic elastase has been determined using 125Iodine-labeled elastin as substrate. Elastin degradation was significantly increased in the presence of 10(-3) M Mg++. If elastin was pre-incubated with 0.5 (w/v) Triton, there was a further increase in elastinolysis to 2.6 times the original rate.     

Immunoperoxidase localization of elastin in the human aorta

Summary Use of the peroxidase-antiperoxidase technique with preliminary trypsinization has allowed immunolocalization of elastin in human aorta. By this method it is possible to utilize formalin-fixed and paraffin-embedded tissue. The advantages in studying a strongly-autofluorescent material are discussed.     

Magnesium enhances human pancreatic elastase digestion of125I-labeled elastin

Summary The effect of some divalent cations, especially Mg⁺⁺, on elastinolysis by porcine or human pancreatic elastase has been determined using¹²⁵Iodine-labeled elastin as substrate. Elastin degradation was significantly increased in the presence of 10^–3 M Mg⁺⁺. If elastin was pre-incubated with 0.5 (w/v) Triton, there was a further increase in elastinolysis to 2.6 times the original rate.     

Immunosuppressive effect of a human serum ferroprotein of hepatic origin, alpha 2 H globulin. Study on blastic transformation of normal lymphocytes in the presence of phytohemagglutinin]

The alpha2 H globulin, a glycoferroprotein, has been found in the serum of patients with malignant diseases. Its level varies according to the evolution of the disease. The activity of this protein has been studied on the normal lymphocytes cultivated in presence of phytohemagglutinin. This study has showed that 5 mug/ml of alpha2 H globulin can inhibit the blastic transformation. The inhibitory effect is proportional to the amount of alpha2 H globulin added as judged by the tritiated thymidin incorporation. The study of morphological aspects of the lymphocytes shows that alpha2 H globulin acts upon the synthesis of the cytoplasmic proteins.     

DNA polymerases in human platelets]

Human platelets have two DNA polymerases. In the mitochondrial-free cytoplasm we have found a DNA polymerase gamma and another DNA polymerase very closely related to the polymerase alpha from animal cells. In the mitochondria only the gamma activity is present.     

Phosphorylase activity of human placenta perfused in vitro: effect of gonadotropins, dibutyryl cyclic AMP and prostaglandins]

The gonadotropic hormones stimulate the glycogen phosphorylase activity of human full-term placenta perfused in vitro. This effect is reproduced by dibutyryl cAMP and PG F2alpha and accompanied by an increased glucose liberation in the perfusion fluid.     

Synthesis of [1-Aib]-angiotensin II, an analogue with higher potency than [1-Asn,5-Val]-angiotensin II

[1-Aib]-angiotensin II was synthesized by Merrifield's solid-phase procedure. The analogue, tested on rabbit aorta strips, showed intrinsic activity alpha E = 1, and when tested on rat blood pressure it gave a pD2 of 8.06; a 3.2 +/- 1.3-fold higher potency than the Ciba-Hypertensin standard.     

Prostaglandin-like substances in Propionibacterium acnes. III. Differential contractile effects on smooth muscle layers of the human utero-tubal junction.

The biological activity of a lipid fraction extracted from P. acnes was tested on isolated smooth muscle strips from the human utero-tubal junction. The bioassay experiments support the concept that prostaglandin-like substances (PLS) occur in P. acnes. However, in the bioassay system used, the effect of PLS was different from that of PGF2 alpha and PGI2 but similar, although not identical, to that of arachidonic acid and PGE2.     

Immunosuppressive effect of a human serum ferroprotein of hepatic origin: alpha-2-HF-globulin]

alpha 2HF, a serum ferroprotein increased during some malignant diseases, displays a suppressive effect on in vitro antibody responses, whether T-independent or T-dependent. This effect is not related to the presence of iron, nor to the macroglobulin nature of the molecule. The immunosuppressive properties of alpha 2 HF might depend on its carbohydrate moeity.     

Chromosome aberrations,micronuclei, and activity of superoxide dismutases in human lymphocytes after irradiation in vitro

The goal of this study was to provide data on the dose-dependent production of dicentrics and micronuclei in human lymphocytes irradiated with 22.6 MeV protons and to estimate the possible contribution of intracellular superoxide dismutases (SOD) to the relative biological effectiveness (RBE) of protons. For the dose-response study, heparinized whole blood of a healthy volunteer was irradiated with protons and X-rays employing radiation doses of 0.5-4 Gy. Three biological endpoints were analyzed: chromosomal aberrations, micronuclei, and specific activity of cytosolic (CuZnSOD) and mitochondrial (MnSOD) superoxide dismutases in harvested human blood cells. Dicentric dose-response curves fit a linear-quadratic form (alpha = 0.094 +/- 0.006, beta = 0.032 +/- 0.001) induced with X-rays and (alpha = 0.119 +/- 0.057, beta = 0.029 +/- 0.014) for 22.6 MeV protons. Protons were more effective than X-rays in producing exchanges, particularly at 0.5 and 1 Gy. In contrast to X-ray irradiated samples where a significant increase in the specific activity of MnSOD was recorded (up to a radiation dose of 1 Gy), irradiation with protons markedly reduced its activity. As a consequence of the reduced activity of MnSOD, the chromosomal dose-response curve became quadratic. The RBE for dicentrics varies with dose (from 2.2 to 1.01) and reduced activity of MnSOD is an important contributor to the RBE of protons. SODs, particularly MnSOD, play an important role in defending DNA from reactive oxygen species. A reduced activity of SOD, particularly MnSOD, is an important contributor to the RBE of protons.     

Influence of beta-aminoproprionitrile (BAPN) on cell growth and elastic fiber formation in cultures of auricular chondrocytes

Auricular chondrocytes isolated from 4-day-old rabbits and grown in vitro for 14 days, proliferated rapidly and produced a conspicuous network of elastic fibers. Beta-aminoproprionitrile (BAPN), which in vivo inhibits cross-linking of elastin, decreased the formation of elastic fibers at a concentration of 10-20 micrograms/ml and prevented formation at 40 micrograms/ml. At a concentration of 5 micrograms/ml only the so-called patches of elastin appeared to be absent. The inhibitory effect of BAPN on cell growth did not exceed 10%, which indicates that BAPN is only slightly harmful to auricular chondrocytes and can safely be used in studies on elastin deposition by these cells in vitro.     

Model of the interaction between trypsin and alpha 2-macroglobulin of porcine serum]

A comparative study of the dissociation into subunits of Porcine alpha2 M, either native or bound to trypsin (Tn), has been carried out in order to determine the modifications of the alpha2 M structure due to the formation of the Tn-alpha2 M complex. Analytical ultra-centrifugation at pH 3.5 shows that the dissociation is smaller when alpha 2 M is bound to trypsin. Electrophoresis in 4% polyacrylamide gels, in presence of 0.1% SDS, of alpha2 M and Tn-alpha2 M incubated in 1% SDS leads to the same conclusion; the enzyme must stabilize the quaternay structure of alpha2 M. In presence of SDS + beta-mercaptoethanol, only a molecular weight (M.W.) 200,000 band is revealed in electrophoresis pattern of native alpha2 M. In the case of reduced Tn-alpha2 M, some other bands of M.W. 100,000, 50,000, 30,000 appear. When trypsin is inactivated by TLCK 100,000 M.W. band is present, accompanied by the 200,000 M.W. band whose intensity is function of the alpha2 M concentration. The 100,000 M.W. band appears therefore characteristic of the formation of the complex which must imply a proteolytic cleavage in the middle of the 100,000 polypeptidic chain of alpha2 M. A model of the complex is proposed in which the enzyme forms a proteic bridge between the two halves of the alpha2 M molecule.     

Calcium dobesilate (Doxium) as a prostaglandin synthetase inhibitor in pregnant human myometrium in vitro

This comparative study on the effect of calcium dobesilate and indomethacin on prostaglandin biosynthesis was performed on microsomal fractions of pregnant human myometrium. Both drugs inhibited prostaglandin synthesis, indomethacin being more potent. Calcium dobesilate inhibited, in a dose-dependent manner, the synthesis of 6-oxo-PGF1 alpha, PGF2 alpha, PGE2 and TXB2. Its inhibitory action is comparable to that of etamsylate.     

Progressive antithrombin activity in rabbit plasma. Role of alpha macroglobulins]

Immunological methods associated with a kinetic study of blood coagulation allowed us to show the influence of the 2 rabbit alpha macroglobulins on the plasma progressive antithrombin activity. This study allowed the authors to discuss the participation of the alpha macroglobulins during the fibrinoformation and to think that these proteins have an immediate antithrombin activity.     

The modulus of elasticity of lobster aorta microfibrils

The presence of elastic fibres in the extracellular matrix (ECM) provides physiologically important elastic properties for many tissues. Until recently, microfibrils, one component of the ECM, were thought primarily to serve as a scaffolding on which elastin is deposited during development to form elaunin fibres [1]. The most prominent protein that forms mammalian microfibrils is fibrillin. It is known that mutations in the fibrillin gene cause a heterogenous connective tissue disease called marfan syndrome [2], so information on mechanical properties of microfibrils or their role in tissue function would be useful. Microfibrils are also found in the ECM of some invertebrate tissues, and there is growing evidence that the protein forming the structure is homologous to mammalian fibrillin [3, 4]. It has been shown that the microfibril-based arterial wall of the lobster has viscoelastic properties [5], and we have now utilized this primitive artery to measure the modulus of elasticity of microfibrils. It is similar to that of the rubber-like protein elastin.     

Altered arterial connective tissue in racing greyhound dogs.

Vascular collagen and elastin contents and the ratio of collagen/elastin (C/E) were studied in racing greyhound dogs, a breed which exhibits increased cardiac output. As compared to mongrel dogs, vascular C/E was lower, suggesting a greater distensibility of vessels as an adaptive response to hemodynamic stress.     

Enhancement of humoral immune response against human lung elastin peptides

Summary When guinea pigs instead of rabbits were used as the host animals, 8–16 times higher antibody titers against human lung elastin peptides were produced with only 1/20 the amount of antigen per unit body weight. This corresponds to a 200-fold enhancement of the immune response.Presented at the 6th Colloquium of the Federation of European Connective Tissue Clubs, Paris, August 28–30, 1978. The data form part of the Ph.D. thesis submitted by T.V. Darnule to the Department of Biology, New York University. Supported in part by NIH Program Project Grant HL15832 and by a Parker B. Francis Fellowship to T.V. Darnule.     

Inhibition of neutrophil-mediated cytotoxicity by alpha 2 macroglobulin

Highly purified human alpha 2 macroglobulin (1.2--10 mg/ml) was shown to inhibit phytohemagglutinin-induced or antibody-induced lysis of chicken erythrocytes by polymorphonuclear neutrophils. Inhibition was not associated with impaired contact between effector and target cells but rather with the antiprotease activity of alpha 2 macroglobulin.