The outcome of Japanese anorexia nervosa patients treated with an inpatient therapy in an internal medicine unit.

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.

Tubulin stimulates adenylyl cyclase activity in rat striatal membranes via transfer of guanine nucleotide to Gs protein.

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.

Effects of ondansetron on electrically evoked contraction in rat stomach fundus: possible involvement of 5-HT2B receptors.

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.

Characterization of the contractile response induced by 5-methoxytryptamine in rat stomach fundus strips.

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.

Effects of dopamine on adenylyl cyclase activity and amylase secretion in rat parotid tissue.

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.

Alpha 1- and beta-adrenergic and muscarinic-cholinergic regulation in the spontaneous beating and Ca2+ oscillations in cultured neonatal rat cardiac myocytes.

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.

Antidepressants directly influence in situ binding of guanine nucleotide in synaptic membrane.

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.

[Improvement of inter-assay for the standardization of PT and TT--clinical significance of local standardization method].

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.

[Effects of epidural analgesia combined with general anesthesia on hemodynamics during neck surgery].

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.

[Disorders of coagulation/fibrinolysis and bleeding tendency].

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.

[Effects of chronic oral administration of ketanserin and trichlormethiazide on blood pressure in stroke-prone spontaneously hypertensive rats].

The present study was undertaken to elucidate the effects of ketanserin and trichlormethiazide on blood pressure in stroke-prone spontaneously hypertensive rats (SHRSP). Oral ketanserin and trichlormethiazide were administered to separate groups for 4 weeks. Combined ketanserin and trichlormethiazide was also administered and the effects observed. 1) In both ketanserin and trichlormethiazide administered groups, significant antihypertensive actions were observed as compared with the non-treated SHRSP. 2) In the ketanserin administered group, a significant increase in water drinking activity was observed, accompanied with an increased urinary volume. 3) In the trichlormethiazide administered group, urinary sodium excretion decreased significantly, accompanied with an increased urinary norepinephrine excretion. 4) Combined administration of ketanserin and trichlormethiazide enhanced the decreases in blood pressure which were produced by single administration of either ketanserin or trichlormethiazide. Moreover, both the increasing tendency in the urinary sodium excretion and the decreasing tendency in the urinary norepinephrine excretion occurred after the combined administration in SHRSP. These findings suggest that ketanserin produces an antihypertensive effect and may act prophylactically for thiazide-induced renal impairment in SHRSP.

Enhancement in beta-adrenergic responsiveness of adenylate cyclase in rat liver after galactosamine administration.

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.

[Effects of ethanol on Gi protein function in rat cerebral cortex of Wistar and Fischer 344 rats: evaluation by low pH treatment].

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.

Functional activity of protein S determined with use of protein C activated by venom activator.

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.

Amidolytic kinetic assay of protein C by selective spectrophotometry in a centrifugal analyzer.

This rapid, simple amidolytic assay of protein C activity in whole plasma involves activation by protein C activator from the venom of Agkistrodon contortrix contortrix (Protac) and use of a Cobas Fara spectrophotometer programmed for kinetic assay. Plasma is incubated with activator venom in the presence or absence of antibody to human protein C in the instrument, chromogenic substrate (S-2366) is added, and the absorbance is measured at 405 nm. The difference between the absorbance of the sample plasma with and without antibody to human protein C correlated well with protein C antigen as assayed by enzyme-linked immunosorbent assay (ELISA) and the Laurell rocket technique in normal subjects, patients being treated with warfarin, and patients with liver cirrhosis or disseminated intravascular coagulation. Our mean value for protein C in normal subjects is 115.9 (SD 16.7)% for amidolytic activity, 103.0 (SD 17.4)% for ELISA, and 97.2 (SD 18.1)% for the rocket technique. The high value for normal subjects presumably includes some nonspecific amidolytic activity activated by the activator venom, as indicated by measurable activity in immuno-depleted protein C-deficient plasma. Within-run and between-run CVs were less than 5% at low, normal, and high concentrations of protein C amidolytic activity.

Antibody studies of factor VIII inhibitor in a case with Waldenström's macroglobulinemia.

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.

A case of hepatocellular carcinoma with marked hyperfibrinogenemia.

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.