RESUMO
When 70-80-g male albino rats eat a diet furnishing daily requirement of valine for optimal growth (70 mumol/g) and all other nutrients ("complete diet"), they gain weight at an average rate of 3.0 g/100 g body wt/day. When valine is removed, they lose weight at an average 2.1 g/100 g body wt/day. The growth retardation is improved or corrected by adding valine to the diet, daily weight gain being proportional to dietary valine content over a range of 0-70 mumol/g. Addition of alpha-ketoisovaleric acid instead of valine to the valine-free diet also improves or corrects the growth failure. Percent efficiency of alpha-ketoisovaleric acid as a substitute for valine was calculated as: 100 x (micromole valine per gram diet required to produce specified growth response)/(micromole alpha-ketoisovaleric acid per gram diet required to produce the same response). Efficiency of the substitution is inversely related to dietary content of the keto analogue, being 80% when diet contains 17.5 mumol/g (molar equivalent of (1/4) the daily requirement of valine), and 37% when diet provides 140 mumol/g (molar equivalent of twice the daily requirement of valine).alpha-Hydroxyisovaleric acid also substitutes for valine. Efficiency of the substitution at the single ration tested, 70 mumol/g diet, is 45%, similar to that for the keto analogue under the same conditions. When [1-(14)C]alpha-ketoisovaleric acid is injected intravenously, 30-80% of the administered radioactivity is exhaled as (14)CO(2) within 24 h. This finding suggests that inefficiency of alpha-ketoisovaleric acid as a substitute for valine results in part from degradation of the keto acid to isobutyric acid by branched chain dehydrogenase-decarboxylase. Oral administration of neomycin, polymyxin, and bacitracin reduces efficiency of alpha-ketoisovaleric acid as a substitute for valine by (1/4)-(1/2). This effect suggests that transamination of the keto acid may be performed in part by gastrointestinal microbes.
Assuntos
Cetoácidos/metabolismo , Valeratos/metabolismo , Valina/metabolismo , Animais , Bacitracina/farmacologia , Peso Corporal/efeitos dos fármacos , Butiratos/metabolismo , Dióxido de Carbono/biossíntese , Radioisótopos de Carbono , Cromatografia em Camada Fina , Descarboxilação , Dieta , Crescimento/efeitos dos fármacos , Absorção Intestinal , Intestino Delgado/efeitos dos fármacos , Cetoácidos/síntese química , Cetoácidos/farmacologia , Espectroscopia de Ressonância Magnética , Síndromes de Malabsorção/metabolismo , Masculino , Neomicina/farmacologia , Pentanóis/síntese química , Pentanóis/metabolismo , Pentanóis/farmacologia , Polimixinas/farmacologia , Ratos , Valeratos/síntese química , Valeratos/farmacologia , Valina/análise , Valina/farmacologiaRESUMO
Although it is a well known fact that hepatocytes are the primary source of plasma proteinase inhibitors, including alpha 1-antichymotrypsin, this protein has also been detected in lung epithelial cells, which may suggest its local production. We have demonstrated that lung-derived epithelial cells are capable of synthesizing high levels of alpha 1-antichymotrypsin. In normal bronchial epithelial cells, as well as in the HTB55 human adenocarcinoma cell line, alpha 1-antichymotrypsin synthesis was under the control of inflammatory cytokines, of which oncostatin M was the most potent stimulator. This finding is consistent with a role for this inhibitor in protecting the lung epithelium from damage by chymotrypsin-like enzymes released from phagocytes such as neutrophils following pathogen invasion.
Assuntos
Citocinas/farmacologia , Pulmão/enzimologia , alfa 1-Antiquimotripsina/metabolismo , Adenocarcinoma/patologia , Brônquios/metabolismo , Dexametasona/farmacologia , Epitélio/enzimologia , Expressão Gênica/efeitos dos fármacos , Humanos , Técnicas In Vitro , Inflamação/fisiopatologia , Oncostatina M , Peptídeos/farmacologia , RNA Mensageiro/genética , Células Tumorais CultivadasRESUMO
Adrenal glands from early, mid, and late fetuses of rabbit, guinea pig, and rat, and from newborn animals of each species, were incubated for 1-4 h with and without 0.1 nM-1 microM ACTH, alpha- or beta-melanocyte-stimulating hormone (alpha MSH or beta MSH). The effects of the peptides were measured on production of glucocorticoids, and on incorporation of labeled thymidine or leucine into DNA or protein, respectively. The findings were similar in all three species. ACTH stimulated synthesis of glucocorticoids throughout fetal life. Potency increased progressively, as reflected by declining minimal effective dose and rising maximal response. In early and mid fetus alpha MSH and beta MSH caused a modest glucocorticoid steroidogenic effect. ACTH and alpha MSH stimulated DNA and protein synthesis in the early and mid fetal gland. alpha MSH was more potent than ACTH in these respects, minimal effective dose being generally 10 times less and maximal response 25-200% greater. The effects diminished or disappeared in the late fetal and newborn gland. These data indicate that alpha- and beta MSH possess steroidogenic or growth-promoting properties, or both, for the fetal adrenal gland.
Assuntos
Glândulas Suprarrenais/efeitos dos fármacos , Hormônio Adrenocorticotrópico/farmacologia , Hormônios Estimuladores de Melanócitos/farmacologia , Glândulas Suprarrenais/embriologia , Glândulas Suprarrenais/metabolismo , Animais , DNA/biossíntese , Feminino , Idade Gestacional , Glucocorticoides/biossíntese , Cobaias , Gravidez , Biossíntese de Proteínas , Coelhos , Ratos , Especificidade da EspécieRESUMO
Patient B. J. with chronic myelocytic leukemia excreted 0.5-1.1 g protein per day in the urine. Gel filtration on Sephadex G-75 showed about one-third of this protein to be in molecular weight range 20,000-40,000 (fraction BJC). BJC, prepared from 9 liters of urine by gel filtration, was chromatographed on carboxymethylcellulose. Two proteins were eluted from the resin in pure form (as shown by zone and immunoelectrophoresis) in yields representing 8 and 3 mg/liter of urine: BJC1 and BJC2. Their amino acid compositions were identical. BJC1 contained 61% carbohydrate (33% hexose, 11% sialic acid, 13% glucosamine, 5% galactosamine). BJC2 contained one-fourth to one-half as much of each carbohydrate. Molecular weight of BJC1 was estimated at 29,000 by gel filtration. Neither glycoprotein reacted with rabbit antiserum to normal human serum.Antiserum to BJC1 was made in the rabbit. Immunoelectrophoresis with this antiserum showed a faint precipitin line, corresponding in mobility to BJC1, in normal human plasma, and a stronger line in most leukemic plasmas. By immunodiffusion, BJC1 was not detectable in normal human urine, but a positive reaction occurred in the following conditions: leukemia, 64-72%; other types of disseminated neoplastic disease, 36-78%; regional ileitis, 45%; ulcerative colitis, 38%; tuberculosis, 33%; during the 1st wk after major surgery, 33%.BJC2 was found in the urine by immunoelectrophoresis in 10% of patients with neoplastic disease and was not observed in urine of other patients or in human plasma. Amino acid composition, carbohydrate content, and antigenic specificity indicate BJC1 is a previously unrecognized member of the system of normal human plasma glycoproteins. Like certain other glycoproteins, its plasma concentration frequently increases in patients with neoplastic disease, chronic inflammatory disease, or tuberculosis and after surgery. Because molecular weight is 29,000, increased plasma concentration readily causes its appearance in the urine.
Assuntos
Glicoproteínas/urina , Leucemia Mieloide/urina , Idoso , Aminoácidos/análise , Proteínas Sanguíneas/análise , Carboidratos/análise , Cromatografia em Gel , Cromatografia por Troca Iônica , Eletroforese em Papel , Eletroforese em Gel de Poliacrilamida , Feminino , Glicoproteínas/análise , Glicoproteínas/isolamento & purificação , Humanos , Soros Imunes , Imunodifusão , Imunoeletroforese , Leucemia Mieloide/sangue , Contagem de Leucócitos , Peso MolecularRESUMO
The specific activity of the branched-chain alpha-keto acid (BCKA) dehydrogenases was measured in normal tissues of the rat, monkey, and man, and in cirrhotic human liver. In the rat, specific activity of the dehydrogenases in liver, kidney, and muscle averaged 33, 26, and 0.4 U/g wet tissue, respectively; proportion of the body's content of the enzyme located in these three organs was 70, 12, and 10%. In the monkey, specific activities in liver and kidney were only one-half to one-third as great as in the rat, whereas activity in muscle was the same; the monkey's body content of dehydrogenase was distributed 50% in liver, 13% in kidney, and 20% in muscle. In man, specific activities in liver and kidney were only 1/15th to 1/25th as great as in the rat, but activity in skeletal muscle was the same. Distribution of the dehydrogenases in man was 30% in liver, 2% in kidneys, and 60% in muscle. In six patients with alcoholic cirrhosis, specific activity of the dehydrogenase in liver was reduced to 20-50% of normal (average, 32%). This reduction may alter the efficiency of BCKA as substitutes for branched-chain amino acids when BCKA are administered orally, but will have little influence on efficiency when they are given intravenously.
Assuntos
Cetona Oxirredutases/metabolismo , Complexos Multienzimáticos/metabolismo , Animais , Haplorrinos , Humanos , Cetoácidos/metabolismo , Rim/enzimologia , Fígado/enzimologia , Cirrose Hepática/enzimologia , Músculos/enzimologia , Ratos , Fatores de TempoRESUMO
Previous studies showed hyperre-sponsiveness to human growth hormone (hGH) in men with myotonic or limb girdle dystrophies (MMD or LGD). Because polyamines may mediate some actions of hGH, we have now investigated polyamine metabolism in these and other dystrophies. Under metabolic balance study conditions, serum and urine levels of putrescine (Pu), spermidine (Sd), spermine (Sm), and cadaverine (Cd) were measured in six normal men (36-44 yr), four men with MMD (38-44 yr), and three men with LGD (30-36 yr), before and during treatment with 0.532 U/kg body wt ((3/4)/d) of hGH. Daily balances of N, P, and K were also monitored. In the normal subjects, hGH did not influence elemental balances or serum and urine polyamines. In MMD, hGH caused significant retention of N, P, and K (P < 0.005). Basal levels of Sm and Cd were significantly elevated above normal (P < 0.005), and Pu, Sm, and Cd increased two- to fourfold above basal during hGH treatment (P < 0.005). In LGD, hGH also caused retention of N, P, and K. Basal levels of nearly all the polyamines (not serum Pu) were significantly above normal in serum and urine (P < 0.05). During hGH treatment, all four polyamines rose significantly above basal (P < 0.005). Serum and urine polyamine levels in five boys with Duchenne muscular dystrophy, age 8-13, did not differ from those in five age-matched normal boys. Skeletal muscle polyamines were measured in five men (31-40 yr) without muscle disease and in three men with LGD (30-38 yr). Average concentrations of Pu, Sd, Sm, and Cd were 46, 306, 548, and 61 nmol/g wet wt in LGD and 1, 121, 245, and 14 in the normal subjects, respectively (P < 0.05 in each instance). Polyamines were determined in skeletal muscle, liver, kidney, and brain of male mice with hereditary muscular dystrophy and in age- and sex-matched normal controls. Pu, Sd, Sm, and Cd levels were two to three times higher than normal in muscle, but did not differ in liver, kidney, and brain. Similar findings were made in male hamsters with hereditary dystrophy and in their controls. The abnormality in hamster muscle polyamines appeared between 1 and 6 wk of age and persisted or intensified until 30 wk. These data reveal abnormalities of polyamine metabolism in men with MMD, in men with LGD, and in mice or hamsters with hereditary muscular dystrophy. The polyamine disorder could be related to dystrophic patients' hyperresponsiveness to hGH.
Assuntos
Hormônio do Crescimento/farmacologia , Distrofias Musculares/metabolismo , Poliaminas/metabolismo , Adolescente , Adulto , Animais , Cadaverina/metabolismo , Criança , Cricetinae , Humanos , Masculino , Camundongos , Distrofias Musculares/tratamento farmacológico , Distrofias Musculares/genética , Distrofia Muscular Animal/metabolismo , Putrescina/metabolismo , Espermidina/metabolismo , Espermina/metabolismo , Fatores de TempoRESUMO
The serum and urine polyamines putrescine, spermidine, and spermine were measured in 112 normal subjects from 0 to 70 yr of age, and in three groups of short children from 7 to 20 yr: 21 growth hormone (GH) deficient patients, 20 normal variant short stature children, and 9 girls with 45, X Turner's syndrome. Urine polyamines were expressed as micromoles per gram of creatinine or per kilogram body weight, and serum polyamines were expressed as nanomoles per milliliter. In normals, the three polyamines were highest in urine and serum at birth. The mean levels declined progressively with age, the rate of change decreasing with age. The mean for the normal subjects, and its 95% confidence and prediction intervals, were estimated from birth to age 70 for each serum and urine polyamine. In GH-deficient children, serum and urine values were significantly lower (P < 0.05) than the age-specific normal values (with the exception of serum spermidine and spermine), averaging 25-55% below normal. This abnormality was corrected during 1 wk of treatment with human GH. In Turner's syndrome, serum and urine values were significantly reduced (P < 0.05), averaging 35-80% below age-specific normals. GH treatment had no corrective effect. In 6 of 20 normal variant short stature children, polyamine levels were significantly (P < 0.01) subnormal, averaging 50-80% below age-specific normals in both serum and urine. Treatment with GH had no corrective effect. These data show that levels of polyamines in serum and urine are correlated with linear growth primarily during the first decade of life. Subnormal polyamine levels are generally associated with growth retardation.
Assuntos
Transtornos do Crescimento/metabolismo , Poliaminas/metabolismo , Adolescente , Adulto , Fatores Etários , Idoso , Criança , Nanismo/metabolismo , Feminino , Hormônio do Crescimento/deficiência , Hormônio do Crescimento/farmacologia , Humanos , Masculino , Pessoa de Meia-Idade , Poliaminas/sangue , Poliaminas/urina , Putrescina/metabolismo , Espermidina/metabolismo , Espermina/metabolismo , Síndrome de Turner/metabolismoRESUMO
22 nonneoplastic, noninflammatory effusions (cirrhosis and congestive heart failure), 12 non-neoplastic inflammatory effusions (tuberculosis, lupus erythematosus, rheumatoid arthritis, and idiopathic pleuropericarditis), and 58 neoplastic effusions (cancer of lung, breast, ovary, and pancreas, and lymphoma) were analyzed by radial immunodiffusion for orosomucoid concentration. The average concentration +/-SE was 35+/-4, 65+/-17, and 130+/-13 mg/100 ml in the three types of effusion, respectively. By gel filtration and ion exchange chromatography, orosomucoid was isolated from 12 nonmalignant and 14 malignant fluids. The orosomucoid preparations reacted as single components in acrylamide gel electrophoresis at pH 9.0, and in immunodiffusion and immunoelectrophoresis against antisera to human serum and to human plasma orosomucoid. In radial immunodiffusion, the slope of the line relating concentration to the square of the diameter of the precipitate area was identical for orosomucoid isolated from normal human plasma and from nonneoplastic effusions, but was subnormal for orosomucoid isolated from neoplastic fluids. All orosomucoid preparations had normal amino acid composition. Orosomucoid from the nonmalignant effusions had normal carbohydrate content. 11 of 14 samples of orosomucoid isolated from neoplastic fluids had abnormalities in carbohydrate composition, consisting of subnormal content of sialic acid (11 of 14), hexose (10 of 14), and hexosamine (3 of 14), and abnormally high content of hexosamine (4 of 14). Discriminant analysis showed that concentration of orosomucoid distinguished between neoplastic and nonneoplastic noninflammatory effusions more effectively than concentration of total protein, albumin, alpha(1), alpha(2), beta, or gamma-globulin.
Assuntos
Líquido Ascítico/análise , Neoplasias , Orosomucoide/análise , Derrame Pleural/análise , Albuminas/análise , Carboidratos/análise , Fenômenos Químicos , Química , Cromatografia em Gel , Eletroforese em Gel de Poliacrilamida , Globulinas/análise , Humanos , ImunodifusãoRESUMO
Cancer-related urinary glycoprotein EDC1 inhibits the action of trypsin and chymotrypsin on casein and synthetic substrates. The amino acid and carbohydrate compositions of EDC1 are different from those reported for pregnancy-related urinary trypsin inhibitors.
Assuntos
Glicoproteínas/urina , Neoplasias/urina , Inibidores da Tripsina/urina , Carboxipeptidases/antagonistas & inibidores , Caseínas , Quimotripsina , Fibrinolisina/antagonistas & inibidores , Humanos , Leucil Aminopeptidase/antagonistas & inibidores , Elastase Pancreática/antagonistas & inibidores , Pepsina A/antagonistas & inibidores , Ligação Proteica , Compostos de Tosil , TirosinaRESUMO
EDC1, a glycoprotein with a molecular weight of 27,500, was purified from the urine of a leukemic patient, and a radioimmunoassay was developed to use as an immunodiagnostic tool for cancer. Previous studies showed that up to 60% of patients with disseminated neoplastic diseases excreted 100 to 500 mg of EDC1 per day. This protein was immunologically related to inter-alpha-trypsin inhibitor (IATI; M.W. 170,000), a glycoprotein normally present in plasma. EDC1, like IATI, inhibited trypsin and chymotrypsin. EDC1 and IATI have now been found to inhibit the incorporation of thymidine into DNA of normal lymphocytes transformed by phytohemagglutinin. In the presence of 1000 micrograms of EDC1 or 300 micrograms of IATI, incorporation of thymidine by cells was totally inhibited. These proteins were not cytotoxic, did not affect transport of thymidine across the membrane, formed no complex with phytohemagglutinin, and did not compete with phytohemagglutinin for its binding sites. It is proposed that EDC1 and IATI may exert this effect by inhibiting a protease required for blastogenesis.
Assuntos
alfa-Globulinas/farmacologia , Glicoproteínas/farmacologia , Ativação Linfocitária/efeitos dos fármacos , Linfócitos/metabolismo , Proteínas de Neoplasias/farmacologia , Inibidores da Tripsina/farmacologia , Sobrevivência Celular/efeitos dos fármacos , Relação Dose-Resposta a Droga , Humanos , Linfócitos/imunologia , Fito-Hemaglutininas/farmacologia , Desnaturação ProteicaRESUMO
Serum-free growth of the human malignant melanoma cell line Hs0294 is associated with production of transforming growth factor-alpha and an autostimulatory melanoma mitogen (melanoma growth-stimulatory activity, MGSA). The transforming activity is characterized by stimulation of anchorage-independent growth of normal rat kidney fibroblasts and competition with 125I-epidermal growth factor for binding to normal rat kidney cells. The second activity, MGSA, stimulates the anchorage-dependent growth of human melanoma cells in serum-free culture medium. When acetic acid extracts of Hs0294 conditioned medium are subjected to Bio-Gel P-30 chromatography followed by reverse-phase high-pressure liquid chromatography, the majority of the transforming growth factor-alpha elutes at 30 +/- 4% acetonitrile, while the major peak of MGSA elutes at 35 +/- 3% acetonitrile. These data indicate that the anchorage-dependent serum-free growth of the Hs0294 human melanoma cell line is apparently dependent upon the autostimulatory melanoma mitogen, MGSA, which is separable from the 125I-epidermal growth factor competing activity produced by these cells.
Assuntos
Substâncias de Crescimento/isolamento & purificação , Melanoma/análise , Peptídeos/isolamento & purificação , Cromatografia em Gel , Cromatografia Líquida de Alta Pressão , Humanos , Melanoma/patologia , Fatores de Crescimento TransformadoresRESUMO
Gel filtration of urine from Patient ED with acute myelocytic leukemia showed a prominent protein peak with elution position corresponding to molecular weights of 20,000 to 35,000. The protein (EDC1) was isolated in pure form by sequential gel filtration and ion-exchange chromatography. Molecular weight of purified EDC1 was 27,000; it contained 27% carbohydrate and was rich in half-cystine (5% of residues). EDC1 was antigenically and chemically distinct from the recognized glycoproteins of normal plasma. With a specific rabbit antiserum and 125l-labeled EDC1, a radioimmunoassay for the glycoprotein was developed. Both noncancer and cancer plasmas contained immunoreactive material. In noncancer plasma, all the immunoreactivity was eluted from Sephadex G-75 and G-200 in position corresponding to molecular weights of 60,000 to 100,000 (Peak 1). In cancer plasma, an additional peak of immunoreactivity was eluted in the position corresponding to EDC1 (M.W., 20,000 to 30,000; Peak 2). Eighty-six % of urines from patients without clinical cancer were nonreactive in radioimmunoassay (less than 0.1 microgram immunoreactive EDC1 per ml); 11 and 3%, respectively, contained immunoreactivity equivalent to 0.1 to 0.9 and 1 to 9 microgram EDC1 per mi, entirely of Peak 1 type. Ninety-one % of urines from patients with disseminated cancer contained immunoreactivity equivalent to 10 to 9,999 microgram EDC1 per ml, primarily of Peak 2 type.
Assuntos
Glicoproteínas/urina , Leucemia Linfoide/urina , Aminoácidos/análise , Ligação Competitiva , Cromatografia em Gel , Epitopos , Humanos , Masculino , Pessoa de Meia-Idade , Peso Molecular , RadioimunoensaioRESUMO
In 1984, we reported that while immunoreactive levels of serum alpha-1 proteinase inhibitor (API) increased significantly in nine patients with advanced solid tumors, the functional activity of the inhibitor, as measured by the serum trypsin inhibitory capacity, did not increase proportionately. This suggested that a portion of the circulating API was functionally inert. We have now assayed immunoreactive titers and trypsin inhibitory capacity of serum API of 49 patients with advanced carcinomas and 27 healthy controls. Immunoreactive levels of API (expressed as percentage of normal pooled serum which was taken as 100%) in cancer subjects were significantly elevated as compared to normals (mean +/- SE: 233 +/- 9.0% versus 102 +/- 2.0%, P less than 0.05). Although the trypsin inhibitory capacity of the cancer group (16.0 +/- 0.9 units/ml) was significantly elevated (P less than 0.05) as compared to normals (9.9 +/- 0.1 units/ml), this increase was less than that in the immunoreactive titer of API, suggesting the existence of functionally inert API in serum. The fraction of API which was functionally active in this group of cancer patients was 71.0 +/- 3.0% which was significantly less than the normal 98.0 +/- 2.0% (P less than 0.05). In 12 patients followed serially, both immunoreactive levels of API and the trypsin inhibitory capacity increased significantly at the time of clinical progression of disease. There was a significant correlation between increasing absolute granulocyte count and increasing trypsin inhibitory capacity (correlation coefficient 0.66; P less than 0.001). Neither disease progression nor increasing granulocyte count, however, was associated with increasing proportion of functionally inactive API. The inactive form of API had the same molecular weight as the native molecule as shown by gel permeation chromatography and sodium dodecyl sulfate-polyacrylamide gel electrophoresis/Western blot analysis of cancer sera. Therefore, the inactive form was not due to a complex between API and a tumor-derived protease or to proteolytic fragmentation of the native API. Elastase inhibitory capacity of cancer sera with subactive API was essentially identical with trypsin inhibitory capacity indicating that the active site methionine was not oxidized in the inert API. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis/Western blot analysis showed that both normal and cancer serum API existed as two mass variants, at Mr 58,000 and 56,000. Both variants formed complexes with elastase and were functionally active.
Assuntos
Proteínas Sanguíneas , Neoplasias/sangue , Cromatografia Líquida de Alta Pressão , Eletroforese em Gel de Poliacrilamida , Humanos , Peso Molecular , alfa 1-AntitripsinaRESUMO
Patient J. B. with metastatic carcinoma of the colon excreted 0.5 to 1.0 g protein daily, about one-third of which was in Molecular Weight Class 30,000 to 60,000. The major component of this class was isolated by gel filtration and ion-exchange chromatography. The purified protein, labeled JBB5, contained about 11% sialic acid, 8% hexose, and 4% hexosamine. Its molecular weight was between 51,000 and 59,000. It did not react detectably with antisera to any of the recognized normal human plasma proteins. A specific antiserum to JBB5 was raised in the rabbit. Urine from 4% of subjects with nonneoplastic illnesses reacted in double immunodiffusion with anti-JBB5. Thirty-three % positive reactions were obtained with urines from patients with advanced neoplastic disease, the percentage varying from 64% in metastatic cancer of the pancreas to 15% in chronic lymphocytic leukemia.
Assuntos
Adenocarcinoma/urina , Neoplasias do Colo/urina , Glicoproteínas/urina , Proteínas de Neoplasias/urina , Adenocarcinoma/imunologia , Adulto , Anticorpos Antineoplásicos , Neoplasias do Colo/imunologia , Reações Cruzadas , Feminino , Glicoproteínas/imunologia , Glicoproteínas/isolamento & purificação , Humanos , Imunodifusão , Leucemia/urina , Leucemia Linfoide/imunologia , Peso Molecular , Metástase Neoplásica , Proteínas de Neoplasias/imunologia , Proteínas de Neoplasias/isolamento & purificação , Neoplasias/imunologia , Neoplasias/urina , GravidezRESUMO
Extracts of conditioned medium (CM) from Hs0294 human malignant melanoma cells stimulate [3H]thymidine incorporation and an increase in cell number in serum-depleted Hs0294 cells. This activity is acid and heat stable, nonproteolytic, protease sensitive, contains disulfide bonds and elutes broadly from a Bio-Gel P-30 column with an approximate molecular weight range of 6,000 to 25,000. Hs0294 CM also stimulates [3H]thymidine incorporation in nonmalignant human nevus cells and normal rat kidney fibroblast cells but not in human fibroblasts. There was only limited competition with 125I-epidermal growth factor in binding assays. Hs0294 CM extracts stimulate anchorage-independent growth in normal rat kidney fibroblast cells in soft agar but not in Hs0294 cells, nevus cells, or human fibroblasts. This second activity elutes from the Bio-Gel P-30 column in two positions with apparent molecular weights of 27,000 and 11,000.
Assuntos
Substâncias de Crescimento/isolamento & purificação , Melanoma/metabolismo , Ágar , Animais , Contagem de Células , Divisão Celular , Linhagem Celular , Meios de Cultura/análise , DNA/biossíntese , Fator de Crescimento Epidérmico/farmacologia , Humanos , Rim/metabolismo , Rim/ultraestrutura , Peso Molecular , Fatores de Crescimento Neural/farmacologia , Nevo/metabolismo , RatosRESUMO
Bovine pituitary fibroblast growth factor (FGF) stimulates the incorporation of [3H]thymidine into DNA in serum-depleted cultures of some but not other human melanoma cells. The melanotic malignant melanoma cell line MIRW exhibited a 40% increase in [3H]thymidine incorporation into DNA and a 48% increase in cell number in response to 3.73 x 10(-9) M FGF. This same concentration of FGF produced a 22% increase in [3H]thymidine incorporation in the melanotic melanoma cell line Hs0294. However, FGF had no effect on the amelanotic melanoma cell line Hs0675, early-passage cultures of a human amelanotic melanoma (W-1), or early-passage cultures of a congenital nevus (N-1).
Assuntos
Melanoma/fisiopatologia , Mitógenos/farmacologia , Peptídeos/farmacologia , Divisão Celular/efeitos dos fármacos , Linhagem Celular , Replicação do DNA/efeitos dos fármacos , DNA de Neoplasias/biossíntese , Fatores de Crescimento de Fibroblastos , Humanos , Cinética , Melanoma/patologiaRESUMO
We reported previously that an antitryptic glycoprotein, EDC1 (Mr 27,500), which is immunologically related to the normal serum proteinase inhibitor, inter-alpha-trypsin inhibitor (IATI), is excreted in large quantities in the urine of metastatic cancer patients. We have now measured immunoreactive titers of urinary EDC1 and five serum proteinase inhibitors (including IATI) in 16 patients with hematological cancers, 9 patients with various solid tumors, and 32 healthy subjects. The mean urinary EDC1 levels were 22-fold greater in all cancer patients as compared to normals [187.0 +/- 136.6 (S.D.) versus 8.4 +/- 8.2 mg/g creatinine; p less than 0.001]. In the cancer group, serum levels of immunoreactive alpha 1-proteinase inhibitor (also called alpha 1-antitrypsin), alpha 1-antichymotrypsin, and C1 inactivator averaged 152, 237, and 165% of the normal values, respectively (p less than 0.01). Immunoreactive alpha 2-macroglobulin levels were unchanged, and immunoreactive IATI levels were depressed (75% of the normals; p less than 0.01). The lower levels of IATI and elevated levels of EDC1 are consistent with the latter being derived from the former. In spite of the increased immunoreactive alpha 1-proteinase inhibitor level, the serum antitryptic capacity of the cancer group averaged only 50% of the normal group (p less than 0.01; range, 5 to 110% of normal average). This suggests that about 70% of the serum alpha 1-proteinase inhibitor in the cancer group is functionally inert.
Assuntos
Neoplasias/sangue , Inibidores de Proteases/sangue , Adolescente , Adulto , Idoso , Feminino , Humanos , Rim/fisiopatologia , Leucemia/sangue , Masculino , Pessoa de Meia-Idade , Neoplasias/urina , Inibidores de Proteases/urina , Valores de ReferênciaRESUMO
The plasma and 24-hr urinary levels of cyclic adenosine 3':5'-monophosphate and of cyclic guanosine 3':5'-monophosphate (cGMP) were determined for 19 healthy normal patients, 54 patients with six types of nonneoplastic diseases (cholelithiasis, peptic ulcer, coronary heart disease, hypertension, regional ileitis, and cirrhosis), and 54 patients with five types of neoplastic disease (cancers of the lung, colon, and breast, acute myelocyte leukemia, and Hodgkin's disease). The cyclic adenosine 3':5'-monophosphate levels of urine and plasma in normal subjects, in noncancer subjects, and in cancer subjects did not differ significantly. The cGMP levels in the noncancer group were similarly unchanged from those in the normal group. However, mean cGMP levels in the urine and plasma of patients with neoplastic diseases were, respectively, 2- and 3-fold greater than the normal values (p less than 0.005 for urine and p less than 0.05 for plasma). Pharmacokinetic studies with [3H]cGMP in nine healthy controls and 15 patients with neoplasia showed that the mean production rate of this nucleotide in patients with metastatic cancer was elevated when compared to normal patients, but many values fell within the normal range. In acute leukemia, the production rate was seven times normal, with four of five patients having values clearly outside the normal range. The plasma clearance rate in patients with neoplasia was not decreased when compared to that in normal patients. It is proposed that an increased production rate, rather than any change in plasma clearance, accounts for the increased levels of cGMP in the plasma and urine of some patients with neoplastic disease.
Assuntos
GMP Cíclico/metabolismo , Neoplasias/metabolismo , AMP Cíclico/metabolismo , GMP Cíclico/sangue , GMP Cíclico/urina , Espaço Extracelular/metabolismo , Humanos , Cinética , Taxa de Depuração MetabólicaRESUMO
S-Adenosylmethionine (AdoMet) is an important biologic methylating agent for nucleic acids, phospholipids, biologic amines, and proteins. Previous studies indicated that hepatic AdoMet synthetase and hepatic levels of AdoMet are subnormal in patients with alcoholic cirrhosis. This abnormality limits the patients' capacity to convert phosphatidylethanolamine to phosphatidylcholine by way of phosphatidylethanolamine-N-methyltransferase (PEMT). Because alcoholic consumption appears to be associated with hepatic hypoxia, we previously measured AdoMet concentration in liver cells under acute hypoxia and found the level to be decreased substantially. In the present study, we determined whether a similar metabolic abnormality was also observed in rats maintained under physiologic hypoxia for 9 days and administered standard rat chow. The study showed that AdoMet levels in hypoxic rat (ave +/- SD) were significantly lower than those in the control (36.8 +/- 11.6 vs. 60.4 +/- 2.3 nmol/g liver; P < 0.05). Also significantly lower in the hypoxic group were the activities of AdoMet synthetase (0.60. +/- 0.07 vs. 0.97 +/- 0.20 U; P < 0.05) and PEMT (26.2 +/- 4.2 vs. 35.6 +/- 5.8 U; P < 0.02). The mRNA levels of AdoMet synthetase also declined in hypoxia indicating that hypoxia may modulate the gene expression of hepatic AdoMet synthetase. Thus, in vivo hypoxia may have an important effect on 1-carbon metabolism.
Assuntos
Hipóxia/metabolismo , Fígado/metabolismo , S-Adenosilmetionina/biossíntese , Animais , Sequência de Bases , Regulação Enzimológica da Expressão Gênica , Fígado/enzimologia , Cirrose Hepática Alcoólica/metabolismo , Masculino , Metionina Adenosiltransferase/análise , Metionina Adenosiltransferase/genética , Metiltransferases/análise , Metiltransferases/genética , Dados de Sequência Molecular , Oxigênio/administração & dosagem , Fosfatidiletanolamina N-Metiltransferase , RNA Mensageiro/análise , Ratos , Ratos Sprague-DawleyRESUMO
EDCl is a novel glycoprotein, mol wt 27,000, isolated in 1976 from leukemic urine. It inhibits the serine proteases trypsin and chymotrypsin and is antigenically related to interalpha trypsin inhibitor (IATI), mol wt 170,000, a normal plasma antiprotease. Since psoriasis is a non neoplastic hyperproliferative state, we have now measured EDCl by a specific radioimmunoassay (RIA) in plasma and urine of 24 untreated psoriatic patients. EDCl was not detectable in normal urine (less than 1 mg/gm creatinine) or plasma (less than 1 mg/L). 55% of psoriatic urine specimens were positive by RIA, containing 8 to 110 mg/gm creatinine. 75% of plasmas were positive, containing 12 to 32 mg/L. Plasma and urine contents of EDCl were significantly (P less than .05) correlated with severity of clinical disease (% skin involved) but not with age, sex, distribution or type of lesion, family history or arthritis.