RESUMEN
OBJECTIVE: To investigate the outcome of Japanese anorexia nervosa (AN) patients who were treated with the standard Japanese inpatient therapy. METHOD: Of the 88 female AN patients treated with our inpatient therapy between January 1997 and December 2002, 67 (76.1%) who agreed to cooperate in this study were assessed by the Global Clinical Score (GCS) at admission and follow-up, 6.3±1.8 years after discharge. Their clinical characteristics at admission and discharge were also examined. RESULTS: Four (6.0%) patients had died before follow-up. BMI was significantly increased during inpatient therapy. At follow-up, excellent, much improved, symptomatic, and poor outcomes on GCS were 57.1%, 14.3%, 14.3% and 14.3%, respectively. Younger age at admission and larger BMI at discharge were significantly associated with a better outcome. DISCUSSION: This study shows the potential for the use of this method for the treatment of AN patients in countries without specialized eating disorder units.
Asunto(s)
Anorexia Nerviosa/terapia , Terapia Cognitivo-Conductual/métodos , Pacientes Internos , Adolescente , Adulto , Factores de Edad , Anorexia Nerviosa/mortalidad , Índice de Masa Corporal , Femenino , Estudios de Seguimiento , Unidades Hospitalarias , Humanos , Medicina Interna , Japón/epidemiología , Factores de Riesgo , Resultado del TratamientoRESUMEN
Corticosteroids are widely used in bronchial asthma, but their mechanism of action is not fully understood. The in vitro studies have proposed that human T helper cells, type 1 (Th1) favor expression of CXCR3, whereas Th2 cells favor CCR4. In this study we investigated whether oral prednisolone modulates the balance of peripheral blood CXCR3+ and CCR4+ T cells. We analyzed the T-cell subsets in 28 patients with stable atopic asthma and 13 normal control subjects before and after 2 wk of treatment with prednisolone, 20 mg/d, or placebo in a randomized, double-blind, parallel group study. The numbers of CXCR3+ and CCR4+ memory T cells were measured with a flow cytometer, and expressed as percentages in CD4+/CD45RO+ memory T cells. In the steroid-treated asthma group, there was a decrease in CCR4+ T cells (from 29.3% to 20.3%, p < 0.0001), and an increase in CXCR3+/ CCR4+ ratio (from 1.86 to 2.89, p = 0.0047), whereas there was no change in CXCR3+ T cells. However, the percentages of CCR4+ cells did not change after steroid therapy in normal control subjects. These results suggest that short-term oral corticosteroid modulates the balances of CXCR3+ and CCR4+ cells in patients with asthma.
Asunto(s)
Antiinflamatorios/administración & dosificación , Asma/sangre , Asma/tratamiento farmacológico , Prednisolona/administración & dosificación , Receptores de Quimiocina/biosíntesis , Linfocitos T/metabolismo , Administración Oral , Adolescente , Adulto , Anciano , Método Doble Ciego , Femenino , Humanos , Masculino , Persona de Mediana Edad , Receptores CCR4 , Receptores CXCR3RESUMEN
Macrothrombocytopenia with leukocyte inclusions is a rare autosomal dominant platelet disorder characterized by a triad of giant platelets, thrombocytopenia, and characteristic Döhle body-like leukocyte inclusions. A previous study mapped a locus for the disease on chromosome 22q12.3-q13.2 by genome-wide linkage analysis. In addition, the complete DNA sequence of human chromosome 22 allowed a positional candidate approach, and results here indicate that the gene encoding nonmuscle myosin heavy chain-A, NMMHC-A, is mutated in this disorder. Mutations were found in 6 of 7 Japanese families studied: 3 missense mutations, a nonsense mutation, and a one-base deletion resulting in a premature termination. Immunofluorescence studies revealed that NMMHC-A distribution in neutrophils appeared to mimic the inclusion bodies. These results provide evidence for the involvement of abnormal NMMHC-A in the formation of leukocyte inclusions and also in platelet morphogenesis.
Asunto(s)
Plaquetas/patología , Genes Dominantes , Leucocitos/ultraestructura , Proteínas Motoras Moleculares , Mutación , Trombocitopenia/genética , Secuencia de Aminoácidos , Mapeo Cromosómico , Cromosomas Humanos Par 22/genética , Análisis Mutacional de ADN , Femenino , Humanos , Cuerpos de Inclusión/ultraestructura , Japón , Masculino , Datos de Secuencia Molecular , Morfogénesis , Cadenas Pesadas de Miosina/genética , Neutrófilos/química , Neutrófilos/ultraestructura , Linaje , Alineación de Secuencia , Eliminación de Secuencia , Homología de Secuencia de Aminoácido , Síndrome , Trombocitopenia/patologíaRESUMEN
The autosomal dominant macrothrombocytopenia with leukocyte inclusions, May-Hegglin anomaly (MHA), Sebastian syndrome (SBS), and Fechtner syndrome (FTNS), are rare platelet disorders characterized by a triad of giant platelets, thrombocytopenia, and characteristic Döhle body-like leukocyte inclusions. The locus for these disorders was previously mapped on chromosome 22q12.3-q13.2 and the disease gene was recently identified as MYH9, the gene encoding the nonmuscle myosin heavy chain-A. To elucidate the spectrum of MYH9 mutations responsible for the disorders and to investigate genotypephenotype correlation, we examined MYH9 mutations in an additional 11 families and 3 sporadic patients with the disorders from Japan. Korea, and China. All 14 patients had heterozygous MYH9 mutations, including three known mutations and six novel mutations (three missense and three deletion mutations). Two cases had Alport manifestations including deafness, nephritis, and cataracts and had R1165C and E1841K mutations, respectively. However, taken together with three previous reports, including ours, the data do not show clear phenotype-genotype relationships. Thus, MHA, SBS, and FTNS appear to represent a class of allelic disorders with variable phenotypic diversity.
Asunto(s)
Plaquetas/patología , Leucocitos/patología , Proteínas Motoras Moleculares , Mutación , Cadenas Pesadas de Miosina/genética , Trombocitopenia/sangre , Trombocitopenia/genética , Asia , Secuencia de Bases , Cartilla de ADN/genética , Genes Dominantes , Genotipo , Humanos , Cuerpos de Inclusión/patología , Datos de Secuencia Molecular , Nefritis Hereditaria/genética , Fenotipo , Polimorfismo Genético , Reacción en Cadena de la Polimerasa de Transcriptasa Inversa , SíndromeRESUMEN
We report a case of Waldenström's macroglobulinemia with prominent bleeding tendency; laboratory investigation revealed an elevated activated partial thromboplastin time. Further laboratory evaluation showed circulating factor VIII anticoagulant, deemed polyclonal IgG, with a titer of 700 Bethesda Units/ml. The factor VIII inactivation kinetics of the patient plasma were identical to those of a type II inhibitor, and the inhibitor was found to recognize the A2 domain of the factor VIII heavy chain. Apparently, paraprotein is not always the cause of reduced activity of coagulation factors in neoplastic dysproteinemias.
Asunto(s)
Autoanticuerpos/sangre , Trastornos de la Coagulación Sanguínea/inmunología , Factor VIII/antagonistas & inhibidores , Inmunoglobulina G/sangre , Macroglobulinemia de Waldenström/inmunología , Anciano , Epítopos , Humanos , Inmunoglobulinas/sangre , Cinética , Masculino , Macroglobulinemia de Waldenström/complicacionesRESUMEN
We performed a nationwide Inter-assay including 112 laboratories for the standardization of prothrombin time (PT) and thrombotest (TT). The data were expressed as seconds, percentile and INR. INR was expressed by 2 methods; Method I (conventional method): INR was expressed using each ISI assigned for reagent or reagent-instrument at the respective laboratories and Method II (local standardization method): INR was expressed using each reference curve created with INR assigned standard plasmas at the respective laboratories. (1) Sample distribution of PT as well as TT was the smallest with the data expressed by Method II followed by Method I and then by percentile. The data expressed by seconds was widely distributed and not useful for the standardization of PT and TT. (2) Even the sample distribution obtained by Method II was dependent on the different ISI of the reagents, as it was found that the larger the ISI of the reagents, the wider the distribution of data. (3) The difference between PT and TT of each test plasma was analysed by t-test. It was found that the difference was insignificant when both data were expressed by Method II, but significant when expressed by Method I, suggesting that PT and TT were interchangeable with the use of Method II. (4) Sample distribution of percentile expression and INR with the use of method II was compared. It was revealed that the sample distribution of INR was smaller than that of percentile. It was concluded that INR expressed by the local standardization method was most useful for the standardization of PT and TT.
Asunto(s)
Tiempo de Protrombina , Humanos , MétodosAsunto(s)
Etanol/farmacología , Transducción de Señal , Adenilil Ciclasas/metabolismo , Etanol/efectos adversos , Proteínas de Unión al GTP/metabolismo , Humanos , Receptores de GABA-A/metabolismo , Receptores de N-Metil-D-Aspartato/metabolismo , Transducción de Señal/efectos de los fármacos , Fosfolipasas de Tipo C/metabolismoRESUMEN
The present report concerns a case of hepatocellular carcinoma (HCC) with marked hyperfibrinogenemia. The plasma fibrinogen level reached as high as 1,704 mg/dl. Since treatment against HCC resulted in reduction of plasma fibrinogen and PIVKA-II, an HCC marker, the hyperfibrinogenemia appears to be related to HCC. Immunohistochemically, the HCC specimen from this patient reacted strongly with antiserum to human fibrinogen, suggesting that the elevated fibrinogen was due to synthesis of this protein by the carcinoma cells, not to decreased fibrinolytic activity.
Asunto(s)
Biomarcadores , Carcinoma Hepatocelular/sangre , Fibrinógeno/análisis , Neoplasias Hepáticas/sangre , Proteínas de Neoplasias/sangre , Adulto , Biomarcadores de Tumor/sangre , Carcinoma Hepatocelular/complicaciones , Resultado Fatal , Fibrinógeno/biosíntesis , Hemorragia Gastrointestinal/etiología , Humanos , Técnicas para Inmunoenzimas , Neoplasias Hepáticas/complicaciones , Masculino , Proteínas de Neoplasias/biosíntesis , Precursores de Proteínas/análisis , Protrombina/análisisRESUMEN
This study examined the effects of ondansetron, an antagonist of the 5-hydroxytryptamine (5-HT3) receptor without 5-HT4 receptor agonistic activity. on electrically evoked contractions and acetylcholine release in rat stomach fundus strips. Ondansetron (10(-8)-10(-4) M) produced a concentration-dependent increase in the magnitude of the electrically evoked contraction, while it inhibited the release of acetylcholine induced by electrical field stimulation. The stimulatory effect of ondansetron (10(-6) M) on electrically evoked contractions was antagonized by yohimbine, a 5-HT2B receptor antagonist, but not by SDZ 205-557 (2-methoxy-4-amino-5-chloro-benzoic acid 2-[diethylamino] ethyl ester), a 5-HT4 receptor antagonist. In the presence of tetrodotoxin, ondansetron (10(-7)-10(-5) M) enhanced the contractions induced by acetylcholine. This stimulatory effect of ondansetron on acetylcholine-induced contractions was antagonized by yohimbine. These data suggest that ondansetron potentiates the contractile response to acetylcholine in the rat stomach fundus through the activation of 5-HT2B receptors.
Asunto(s)
Acetilcolina/metabolismo , Ansiolíticos/farmacología , Fundus Gástrico/efectos de los fármacos , Contracción Muscular/efectos de los fármacos , Ondansetrón/farmacología , Receptores de Serotonina/efectos de los fármacos , Animales , Estimulación Eléctrica , Fundus Gástrico/metabolismo , Fundus Gástrico/fisiología , Hidrógeno/metabolismo , Masculino , Ratas , Ratas Wistar , Receptores de Serotonina/metabolismoRESUMEN
This study examined effects of 5-methoxytryptamine (5-MOT), an agonist at 5-HT4 and 5-HT2B receptors, on the contractile response and acetylcholine release in rat stomach fundus strips. 5-MOT (10(-9)-10(-5) M) produced a concentration-dependent increase in the contraction, while it evoked acetylcholine release in a 'bell-shaped' concentration-dependent manner. Atropine reduced 5-MOT (10(-8)-10(-6) M)-induced contractions, but it had little effect on the contractions evoked by higher concentrations. 5-MOT-induced contraction and acetylcholine release were inhibited by SDZ 205-557 (2-methoxy-4-amino-5-chloro-benzoic acid 2-[diethylamino] ethyl ester), a 5-HT4 receptor antagonist. In the presence of atropine, both SDZ 205-557 and yohimbine, a 5-HT2B receptor antagonist, inhibited the contraction. In the presence of tetrodotoxin, the contraction was inhibited by yohimbine, but not by SDZ 205-557. These results suggest that the contractile action of 5-MOT in rat stomach fundus involves atropine-sensitive and atropine-resistant components. The sensitive contraction appears to be mediated through 5-HT4 receptors located on cholinergic neurons, whereas the resistant contraction is mediated through 5-HT4 receptors located on non-cholinergic neurons and through 5-HT2B receptors.
Asunto(s)
Contracción Muscular/efectos de los fármacos , Serotonina/farmacología , Estómago/efectos de los fármacos , Ácido 4-Aminobenzoico/farmacología , 5-Metoxitriptamina/farmacología , Acetilcolina/metabolismo , Animales , Atropina/farmacología , Relación Dosis-Respuesta a Droga , Estimulación Eléctrica , Fundus Gástrico , Técnicas In Vitro , Masculino , Ratas , Ratas Wistar , Estómago/fisiología , Yohimbina/farmacología , para-AminobenzoatosRESUMEN
alpha 1- and beta-adrenergic and muscarinic-cholinergic regulation in spontaneous beating and Ca2+ oscillations in neonatal rat cardiac myocytes at day 6 of culture was investigated. The spontaneous beating in myocytes decreased in the presence of 10 microM norepinephrine (NE). This negative chronotropic action was antagonized by prazosin. Carbachol (CCh) also showed negative chronotropic action which was inhibited by atropine. On the other hand, isoproterenol (ISP) increased the beating rate which was antagonized by propranolol. NE increased inositol phosphate formation whereas CCh and ISP did not. NE and CCh suppressed the frequency of the spontaneous Ca2+ oscillations but ISP increased. The present results suggest that alpha 1-adrenergic and muscarinic receptors regulate chronotropism to be negative whereas beta-adrenoceptor regulates chronotropism to be positive in cultured neonatal rat cardiac myocytes.
Asunto(s)
Calcio/metabolismo , Frecuencia Cardíaca/fisiología , Receptores Adrenérgicos alfa/fisiología , Receptores Adrenérgicos beta/fisiología , Receptores Muscarínicos/fisiología , Adrenérgicos/farmacología , Animales , Animales Recién Nacidos , Atropina/farmacología , Carbacol/farmacología , Células Cultivadas , Depresión Química , Frecuencia Cardíaca/efectos de los fármacos , Fosfatos de Inositol/metabolismo , Isoproterenol/farmacología , Agonistas Muscarínicos/farmacología , Norepinefrina/farmacología , Prazosina/farmacología , Propranolol/farmacología , Ratas , Ratas Wistar , Receptores Adrenérgicos alfa/efectos de los fármacos , Receptores Adrenérgicos beta/efectos de los fármacos , Receptores Muscarínicos/efectos de los fármacosRESUMEN
Previous studies of rat cerebral cortex and rat C6 glioma cells have demonstrated that dimeric tubulin is capable of activating the G proteins Gs and Gil via transfer of guanine nucleotide from tubulin to Gs alpha and Gil alpha. To provide further information regarding cytoskeletal modulation of adenylyl cyclase, the present study examined effects of tubulin on the activation of the enzyme in rat striatal membranes. Tubulin, prepared from rat brain by polymerization with the hydrolysis-resistant GTP analog 5'-guanylylimidodiphosphate (GppNHp) caused significant activation of adenylyl cyclase by approximately 130%. Furthermore, tubulin-GppNHp activated SKF 38393-sensitive adenylyl cyclase and potentiated forskolin-stimulated activity of the enzyme. When tubulin, polymerized with the hydrolysis-resistant photoaffinity GTP analog [32p]p3 (4-azidoanilido)-p1-5'-GTP ([32P]AAGTP), was incubated with striatal membranes, AAGTP was transferred from tubulin to Gs alpha as well as Gi alpha with the extents of nucleotide transfers being 7.6 +/- 0.8% and 17.8 +/- 1.4% of AAGTP originally bound to tubulin, respectively. These results indicate that, in rat striatum, the tubulin dimer participates in the stimulatory regulation of adenylyl cyclase by transferring guanine nucleotide to Gs alpha, supporting the hypothesis that tubulin contributes to the regulation of neuronal signal transduction.
Asunto(s)
Adenilil Ciclasas/agonistas , Cuerpo Estriado/efectos de los fármacos , Nucleótidos de Guanina/metabolismo , Proteínas de la Membrana/metabolismo , Proteínas del Tejido Nervioso/metabolismo , Tubulina (Proteína)/farmacología , Animales , Colforsina/farmacología , Cuerpo Estriado/enzimología , Sinergismo Farmacológico , Proteínas de Unión al GTP/metabolismo , Guanilil Imidodifosfato/farmacología , Masculino , Ratas , Ratas Wistar , Estimulación Química , Tubulina (Proteína)/químicaRESUMEN
Several previous studies have shown that dopamine causes amylase secretion from rat parotid tissue. However, the mechanism of this dopamine action is still unclear. The present study was designed to characterize dopamine action in rat parotid gland tissue by examining the effects of dopamine on cyclic AMP accumulation, adenylyl cyclase activity, and amylase release. Dopamine significantly enhanced accumulation of cyclic AMP in parotid slices and stimulated adenylyl cyclase activity in parotid membrane preparations. It also significantly stimulated amylase release from parotid slices. The stimulatory effects of dopamine on cyclic AMP accumulation, adenylyl cyclase activity, and amylase release were effectively blocked with propranolol, a beta-adrenergic antagonist, but not by either SCH 23390, a preferential D1 antagonist, or butaclamol, a preferential D2 antagonist. No substantial specific binding sites for D1 receptors were detectable by [3H]SCH 23390 binding in parotid membranes. These results suggest that the stimulatory effect of dopamine on amylase secretion in rat parotid tissue is not mediated through specific D1 dopamine receptors but rather through beta-adrenergic receptors.
Asunto(s)
Dopamina/farmacología , Glándula Parótida/efectos de los fármacos , Glándula Parótida/fisiología , Adenilil Ciclasas/metabolismo , Amilasas/metabolismo , Animales , Benzazepinas/farmacología , Butaclamol/farmacología , AMP Cíclico/metabolismo , Dopamina/fisiología , Antagonistas de Dopamina/farmacología , Técnicas In Vitro , Masculino , Glándula Parótida/enzimología , Propranolol/farmacología , Ratas , Ratas Wistar , Receptores Adrenérgicos beta/efectos de los fármacos , Receptores Adrenérgicos beta/fisiología , Receptores Dopaminérgicos/análisis , Estimulación QuímicaRESUMEN
The liver produces almost all the factors involved in coagulation and fibrinolysis. The hepatic reticuloendothelial system also plays an important role in disposing activated coagulation/fibrinolysis-related factors and inhibitors. In liver cirrhosis, the production of coagulation/fibrinolysis-related factors is diminished, as well as its inhibitors. Moreover, the hematological profile is complicated by a decreased rate of clearance, thrombocytopenia induced by hypersplenism, and some qualitative abnormalities of the coagulation factor. Hence, treatment for bleeding tendency observed in cirrhotic patients is diverse.
Asunto(s)
Trastornos de la Coagulación Sanguínea/etiología , Fibrinólisis , Trastornos Hemorrágicos/etiología , Cirrosis Hepática/complicaciones , Antitrombina III/metabolismo , Factores de Coagulación Sanguínea/biosíntesis , Fibrinógeno/metabolismo , Humanos , Cirrosis Hepática/sangre , Plasminógeno/metabolismo , Proteína C/metabolismoRESUMEN
The present study examined the in vitro effects of antidepressants on functional photoaffinity labeling of GTP binding protein. Saturation binding studies were performed by incubating membranes with increasing concentrations of [32P]-AAGTP, followed by UV irradiation and SDS-PAGE. The specifically bound isotherms for each of the G proteins studied showed characteristics of a one site model. Scatchard analysis revealed increases in the Bmax and Kd of AAGTP binding for each of the G proteins (especially stimulatory G proteins) with the addition of antidepressants such as amitriptyline, clomipramine, desipramine and mianserin but not with monoamine oxidase inhibitor, antipsychotics and axiolytics. These results suggested that drugs having antidepressive properties may directly affect G protein, especially Gs protein.
Asunto(s)
Antidepresivos/farmacología , Proteínas de Unión al GTP/metabolismo , Nucleótidos de Guanina/metabolismo , Membranas Sinápticas/efectos de los fármacos , Membranas Sinápticas/metabolismo , Marcadores de Afinidad/metabolismo , Animales , Azidas/metabolismo , Electroforesis en Gel de Poliacrilamida , Guanosina Trifosfato/análogos & derivados , Guanosina Trifosfato/metabolismo , Cinética , Masculino , Radioisótopos de Fósforo , Unión Proteica/efectos de los fármacos , Ratas , Ratas Wistar , Especificidad por SustratoRESUMEN
The present study was designed to provide further information regarding the effect of ethanol on the function of inhibitory GTP-binding (Gi) protein. To eliminate the Gs function in the regulation of adenylyl cyclase, cerebral cortex membranes from Wistar or Fischer 344 rats were pretreated at pH 4.5 ("low pH"), whereupon Gpp(NH)p-dependent inhibition of forskolin-stimulated adenylyl cyclase was examined. In the membranes from Wistar rats, ethanol (100 or 250 mM) resulted in left-shifted Gpp(NH)p inhibition curves and reduced IC50 values for Gpp(NH)p in an ethanol dose-dependent manner. In contrast, ethanol exhibited no effect on the Gpp(NH)p-dependent inhibition of the enzyme or on IC50 values for Gpp(NH)p in the membranes from Fischer 344 rats. These results are consistent with the idea that ethanol, in vitro, enhances the Gi function in the cerebral cortex of Wistar rats, and that ethanol's effect on Gi protein in the cerebral cortex is different in Wistar and Fischer 344 rats.
Asunto(s)
Corteza Cerebral/efectos de los fármacos , Etanol/efectos adversos , Proteínas de Unión al GTP/fisiología , Animales , Corteza Cerebral/metabolismo , Proteínas de Unión al GTP/efectos de los fármacos , Concentración de Iones de Hidrógeno , Masculino , Ratas , Ratas Endogámicas F344 , Ratas WistarRESUMEN
The aim of the present study was to investigate the effect of epidural analgesia combined with general anesthesia on hemodynamics. Thirty patients undergoing surgery for the treatment of cancer of the neck were studied. The patients were divided into two groups of those who received epidural analgesia combined with general anesthesia group (Group 1) and those with general anesthesia alone (Group 2). Blood pressure was not different between the groups. But heart rate and rate pressure products in Group 1 were significantly lower than those of Group 2. CVP in Group 1 increased significantly to 10.1 +/- 2.9 mmHg during surgery from 6.8 +/- 1.8 mmHg at the beginning of the surgery. There was no difference in intraoperative blood loss and the amount of fluid infused between the two groups. These results suggest that epidural anesthesia combined with general anesthesia is effective to stabilize hemodynamics during cervical surgery, but we have to be careful about using local anesthetics during long cervical procedures, because it increases CVP which might result from the depression of cardiac function.
Asunto(s)
Analgesia Epidural , Anestesia General , Neoplasias de Cabeza y Cuello/cirugía , Hemodinámica/fisiología , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Estudios RetrospectivosRESUMEN
In rat liver at 48 hours after galactosamine (GalN) treatment, isoproterenol-induced accumulation of cyclic AMP and activation of adenylate cyclase (AC) were significantly enhanced. AC activity stimulated with GppNHp or NaF was also augmented in membranes from GalN-treated rats. However, the MnCl2-stimulated or forskolin-stimulated activity in GalN-treated rats did not differ from the control. These data suggest that the beta-adrenergic responsiveness of AC in rat liver is potentiated, possibly due to the enhanced function of the Gs protein, during the process of regeneration after GalN treatment.
Asunto(s)
Adenilil Ciclasas/metabolismo , AMP Cíclico/metabolismo , Galactosamina/farmacología , Hígado/enzimología , Receptores Adrenérgicos beta/efectos de los fármacos , Animales , Intoxicación por Tetracloruro de Carbono/metabolismo , Activación Enzimática , Galactosamina/administración & dosificación , Isoproterenol/farmacología , Hígado/efectos de los fármacos , Masculino , Ratas , Ratas EndogámicasRESUMEN
A new screening procedure, an easy and specific assay for determining functional Protein S activity, has been developed, with use of Protein C activated by venom activator (Protac). Purified Protein C (100% amidolytic activity) was activated by venom activator (6 units/mL). To a mixture of 100 microL of Protein S-deficient plasma, 20 microL of sample plasma, 100 microL of cephalin (Actin), and 20 microL of activated Protein C we added 100 microL of 25 mmol/L CaCl2 solution and measured the clotting time with a KC 10 coagulometer. The functional Protein S activity correlated well with the concentrations of Protein S antigen measured by enzyme immunoassay and the Laurell rocket technique (r = 0.810 and 0.850, respectively) in normal subjects, patients with myocardial infarction undergoing warfarin therapy, and patients with liver cirrhosis.