c-kit Mutation in generalized lentigines associated with gastrointestinal stromal tumor.

We describe the case of a 59-year-old Japanese woman presenting with generalized lentigines without systemic anomalies. She had a medical history of gastrointestinal stromal tumors (GISTs), in which gain-of-function mutations of the c-kit gene had recently been found. We detected a point mutation at codon 557 in exon 11 of leukocyte DNA from the patient. The stem cell factor-type III receptor tyrosine kinase pathway plays important roles in the regulation of melanocyte proliferation and differentiation. We speculate that the generalized lentigines of the patient may be caused by melanocyte proliferation due to the c-kit gene mutation.

Effects of heart rate and right ventricular pressure on right coronary arterial flow and its systolic versus diastolic distribution in a variety of congenital heart diseases in children.

BACKGROUND AND METHODS: In order to elucidate the underlying adjusting mechanism of human right coronary arterial (RCA) flow to increased right ventricular pressure (RVP) in children, we recorded RCA flow velocity in 24 pediatric cardiac patients at the orifice of its main trunk at the time of heart catheterization using the Doppler guidewire. RESULTS: The ratio of diastolic flow (DF)/total flow (TF), or the proportion of the DF time integral over a total of one cardiac cycle, had a negative correlation with heart rate (HR; r = -0.58, n = 11) in children with normal right ventricular systolic pressure (RVSP; RVSP < 35 mmHg). In contrast, the DF/TF ratio had a good correlation (r = 0.88, n = 24) with RVSP in all patients under study. The ratio of diastolic area (DA)/total area (TA), defined as the ratio of an area encircled by the aortic pressure curve above and the RVP curve below for diastole, over a total of one cardiac cycle, representing the overall effect of both HR and transcoronary pressure difference, also correlated well (r = 0.89, n = 24) with DF/TF. Total volume flow of the RCA also increased (r= 0.76, n = 24) with increases in RVSP, first by an increase in flow velocity through the RCA, during both systole and diastole, then by widening of the RCA lumen at very high pressures. These changes were initially more dependent on diastole with increasing RVSP because: (i) of a more marked augmentation of flow velocity in diastole compared with systole; and then (ii) of a significant decrease in flow velocity in systole at very high pressures. CONCLUSIONS: We clarify how the RCA manages to increase flow through it at different HR as a function of chronic RVP overload in pediatric cardiac patients.

Clinical features of paradiverticulitis.

Colonic diverticula have been generally accepted as a source of massive hemorrhaging. Little is known, however, about fecal occult blood loss from colonic diverticula and diverticulosis. We retrospectively investigated the possibility of minor bleeding in 737 diverticula cases diagnosed from April 1989 to May 1994. Thirty-seven cases (5%) of diverticula are explained as the sources of positive occult blood testing ascertained clearly by colonoscopy. These divide into three types: (1) from intradiverticular bleeding (intradiverticulitis), (2) from peridiverticular bleeding (peridiverticulitis), and (3) from interdiverticular colonic mucosal erosion (interdiverticulitis). These findings account for the occult blood loss that we call paradiverticulitis. The two-year prospective study found 67 cases (11.3%) of paradiverticulitis in 595 diverticula cases. We concluded that paradiverticulitis is one of the causes of positive occult blood tests.

Effects on gastric circulation of treatment for portal hypertension in cirrhosis.

We evaluated the gastric circulatory effects of the type of treatment administered for portal hypertension. Of 14 patients with cirrhosis, seven received a transjugular intrahepatic portosystemic shunt (TIPS; group T) and seven received percutaneous transhepatic portographic embolization (PTPE; group P). Patients were evaluated over the course of one year. After treatment, portal venous pressure was significantly reduced from 39 +/- 6 cm H2O to 32 +/- 5 (P < 0.001) in group T and was significantly elevated from 29 +/- 10 to 33 +/- 8 (P < 0.05) in group P. The portal flow velocity (Vmean) was significantly higher in group T vs group P (P < 0.0001). The congestion index was significantly lower in group T than in group P (P < 0.0001). The gastric mucosal blood flow was increased in group T but was unchanged in group P. Esophageal varices showed some improvement in both groups, but the portal hypertensive gastropathy was improved only in group T. These findings help to explain the differing effects on the gastric circulation related to the type of treatment used for portal hypertension.

Regulation of hepatic thrombopoietin production by portal hemodynamics in liver cirrhosis.

OBJECTIVE: This study was designed to clarify how thrombopoietin (TPO) functions in and, to some extent, causes thrombocytopenia complicating liver cirrhosis and portal hypertension. METHODS: Our study population consisted of 19 cirrhotic and six noncirrhotic patients who underwent percutaneous transhepatic portography (PTP) and hepatic venography. RESULTS: The platelet counts of the cirrhotic patients were significantly lower than those of the noncirrhotic patients (8.7 +/- 4.1 vs 17.4 +/- 7 x 10(4)/microl; p < 0.01). The flow direction in the splenic vein was confirmed by PTP. Ten hepatofugal and nine hepatopetal flow directions in the splenic vein were noted among the cirrhotics. The hepatofugal group showed lower portal venous pressure (20 +/- 10 vs 32 +/- 4 cm H2O; p < 0.01) than the hepatopetal group and had a higher incidence of hepatic encephalopathy (six of 10 vs zero of nine; p < 0.01). The hepatic vein-portal difference in TPO did not differ substantially between the cirrhotics and noncirrhotics (0.12 +/- 0.04 vs 0.24 +/- 0.07 fmol/ml). Comparisons of this value among the three groups showed the TPO difference to be lowest in the hepatofugal group (hepatofugal: 0.04 +/- 0.03, hepatopetal: 0.21 +/- 0.07, noncirrhotic: 0.24 +/- 0.07; p < 0.05). CONCLUSIONS: Our findings suggest that TPO production in the cirrhotic liver is regulated by the portal blood supply to the liver. Thus, portal hemodynamics may play a critical role in the development of thrombocytopenia.

[A long-term histological prognosis after IFN therapy for chronic hepatitis C].

In 72 chronic hepatitis C patients treated with IFN, liver biopsies were performed before and after IFN treatment, and histological changes were examined. In complete responders, the grading score improved significantly since 6-mo after IFN treatment, and the staging score improved significantly since 12-mo after IFN treatment. In long-term responders underwent a following liver biopsy 4-5 yr after IFN treatments; 5 had normal liver histology. In partial responders, both the grading and staging score did not improve significantly. But, in the cases who had sustained normalization of ALT levels after IFN treatment, both the grading and staging score improved after treatment. In non-responders, both the grading and staging score did not improve significantly.

[Interferon treatment for chronic hepatitis C--assessment of 3 regimens in patients received more than 500MU interferon treatment and their effect predictive factors for interferon treatment using multivariate analysis with the logistic regression model].

Chronic hepatitis C patients (n = 115) were treated with interferon (IFN). Total dose employed was more than 500 MU. The response rate was assessed among the three treatment groups: 2W continuous+TIW, 4W continuous+TIW, 8W continuous+TIW. The IFN treatment effect predictive factors were also assessed. Complete response (CR) rate, CR with serum HCV-RNA disappearance rate, responders' histology activity index score changes between before and after treatment, and responders' hepatocytes HCV-RNA disappearance rate did not differ among the three treatment regimens. CR to IFN treatment was dependent on serum HCV-RNA and HCV serotype. Patients of low serum HCV-RNA and serotype II were responsive to IFN treatment.

Properties of single potassium channels in guinea pig hepatocytes.

The patch-clamp technique of cell-attached and inside-out configurations was used to study the single potassium channels in isolated guinea pig hepatocytes. The single potassium channels in isolated guinea pig hepatocytes were recorded at different K+ concentrations. A linear single-channel current-voltage relationship was obtained at the voltage range of -80 to -20 mV with slope conductance of 70 +/- 6 pS (n = 10). Under symmetrical high K+ concentration of 148 mM in the cell-attached patch membrane, the I-V curve exhibited a mild inward rectification at potentials positive to + 20 mV. The values of reversal potential was +5 +/- 2 mV (n = 10). When the external potassium concentration ([K+]o) was decreased to 74 mM and 20 mM, the slope conductance was decreased to 48 +/- 2 pS (n = 4) and 24 +/- 3 pS (n = 3), respectively. The reversal potential was changed by 58 mV for a tenfold change in [K+]o, indicating that this channel was highly selective for K+. Open probabilities (Po) of the channel were 73-93% without apparent voltage dependence. The distributions of open time of the channels were fitted to two exponentials, while those of closed time were fitted to three exponentials, exhibiting no voltage dependence. The success rate of K+ channel activity to be recorded was 28% at room temperature, and there were no increases in the success rate nor in the channel opening probabilities at a temperature of 34-36 degrees C. Po in inside-out patches was not changed by application of 1 microM Ca2+ nor 1 mM Mg2+ to the internal side of patch membranes. It is concluded that a novel type of the K+ channels in guinea pig hepatocytes had different properties of slope conductance, channel kinetics, and sensitivity to [Ca2+]i, from those in other species.

[Therapeutic potential of sparfloxacin for preventing mycobacterial infections].

We studied the therapeutic potential of utilizing sparfloxacin (SPFX), a newly developed quinolone, to prevent various mycobacterial infections. The in vitro activity of SPFX as a preventive agent for various mycobacteria was determined using the actual count method on Ogawa egg medium. The minimal inhibitory concentrations (MICs) of SPFX were as follows: ofloxacin-sensitive M. tuberculosis, 0.16-0.32 microgram/ml; ofloxacin-resistant M. tuberculosis, 0.63-2.5 micrograms/ml; M. avium; 0.63-10 micrograms/ml (MICs were equal or less than 1.25 micrograms/ml in seven out of 11 strains); M. intracellulare, 2.5-10 micrograms/ml (MICs were equal or more than 10 micrograms/ml in 17 out of 23 strains); M. kansasii, < or = 0.08-0.16 microgram/ml; M. fortuitum, < or = 0.08 microgram/ml; M. chelonae subsp. abscessus, > 10 micrograms/ml; M. chelonae subsp. chelonae, 0.63 microgram/ml; M. scrofulaceum, < or = 0.08 microgram/ml; M. nonchromogenicum, 1.25 micrograms/ml; M. xenopi, < or = 0.08 microgram/ml; M. gordonae, < or = 0.08 microgram/ml. The average serum concentrations of SPFX during the period of multiple oral administration (200 mg once a day) were 0.35 +/- 0.16 microgram/ml before administration, 0.67 +/- 0.32 microgram/ml after one hour, 1.13 +/- 0.21 microgram/ml after two hours, 1.27 +/- 0.32 microgram/ml after four hours and 1.31 +/- 0.34 micrograms/ml after six hours. These results indicate that SPFX has a strong therapeutic potential to prevent infections due to M. tuberculosis, M. kansasii, M. fortuitum, M. chelonae subsp. chelonae, M. scrofulaceum, M. xenopi and M. gordonae. Moreover, it may be expected to be a promising agent against infections due to ofloxacin-resistant M. tuberculosis, M. avium and M. nonchromogenicum.

A study of endotoxaemia in patients with primary biliary cirrhosis.

Hepatic reticuloendothelial function may be diminished in patients with primary biliary cirrhosis. Endotoxin concentrations in peripheral blood and in the superior mesenteric vein were measured, by the beta-glucan sensitive, factor-free, endotoxin-specific limulus assay, in patients with primary biliary cirrhosis and liver cirrhosis (non-PBC cirrhosis). Endotoxaemia was detected in the peripheral blood of seven out of nine patients (78%) with asymptomatic primary biliary cirrhosis, but in only two out of thirteen patients (15%) with non-PBC cirrhosis. The endotoxin level was significantly higher in the earlier stages of primary biliary cirrhosis than in the later stages (P < 0.05). The endotoxin level in superior mesenteric vein blood was significantly lower in patients with primary biliary cirrhosis than in patients with non-PBC cirrhosis. Peripheral endotoxaemia in patients with primary biliary cirrhosis may be due to the diminished capacity of the hepatic reticuloendothelial system, for phagocytosis of endotoxin.

[Henoch-Schönlein purpura associated with pulmonary tuberculosis].

A 34-year-old man was admitted to our hospital because of cough and fever. Chest radiograph showed multiple cavities mainly on the right lung. His sputum was positive for acid-fast bacilli on smear, and he was treated with daily isoniazide, rifampicin and streptomycin. Antituberculous treatment was successfully performed, so acid-fast bacilli of his sputa disappeared on smear and culture. Five months later, he developed a purpuric lesions over both legs accompanied by low grade fever and arthralgia, but proteinuria and abdominal pain could not be observed. Laboratory findings showed a normal platelet count and a normal bleeding time. High levels of serum IgG, IgA, C3 and C4 were evident. ASLO and ASK titer were elevated and they markedly increased within two weeks. A direct invasion of the vessel wall by tubercle bacilli is deniable because antituberculous treatment was successfully continued. Henoch-Schönlein purpura was diagnosed judging from these findings. High levels of ASLO and ASK suggest a preceding streptococcal infection for developing purpura but a possible infectious focus could not be identified. He was treated with 15 mg of prednisolone daily for two weeks and the lesion was subsided. The effect of prednisolone suggests that a subsequent antigen-antibody interaction caused by a streptococcal infection may participate in the formation of the purpura.

[Nutritional assessment and rapid turnover protein in the elderly].

Nutritional assessment with respect to rapid turnover protein (RTP) in the elderly was performed in 22 outpatients (7 males and 15 females, mean age 77 years) who suffered from slight illnesses such as mild hypertension and osteoporosis. Their nutritional parameters were almost within the normal range, judging from body mass index (BMI) and their plasma levels of total protein, albumin, lipids and RTP (transferrin, pre-albumin and retinol-binding protein). Statistically, BMI had a significantly positive correlation with subcutaneous fats and total protein and had a significantly negative correlation with HDL-cholesterol. Retinol-binding protein had a significantly positive correlation with prealbumin, ferritin and vitamin A. Conversely, in 12 bedridden patients the nutritional status was very poor, with their plasma levels of the above markers being significantly lower than those of the controls. We administered an enteral diet (EWH330) to 10 bedridden patients in an effort to restore their nutritional status. As a result, their nutritional parameters, especially RTP showed a significant increase after 4 and 8 weeks of the administration of EWH330. These results suggest that RTP is a very sensitive parameter of nutritional assessment in the elderly.

[A case of pulmonary atypical mycobacteriosis complicated with aplastic anemia, treated with surgical resection and postoperative sparfloxacin].

A few weeks after treatment with INH, RFP and SM, severe aplastic anemia developed in a 49-year-old man with pulmonary atypical mycobacteriosis due to M. kansasii. All drugs were discontinued immediately after bone marrow examination revealed severely hypoplastic marrow, but pancytopenia continued for several months. Although M. kansasii was sensitive to other drugs including CS and TH, these two drugs were also discontinued because of their respective psychiatric and hepatic adverse effects. Ofloxacin (OFLX), to which M. kansasii was sensitive, was administered without clinical improvement and complete resistance to OFLX developed after several months treatment. Right upper and middle lobectomy and S6 partial lobectomy was performed, and postoperative sparfloxacin (SPFX) administration resulted in cure of the disease. Drug sensitivity testing revealed that the organism had acquired resistance to OFLX, but was still sensitive to SPFX. Thus, SPFX appears to be an useful drug for the treatment of atypical mycobacteriosis.

[A case of occupational asthma caused by arrowhead scale in mandarin orange-worker].

A 50-year-old woman with occupational asthma, whose attacks were provoked by inhalation of Arrowhead scale (Unapsis Yanonensis Kuwana) attached to the leaves of mandarin oranges is reported. She experienced asthmatic attacks while picking leaves and harvesting mandarin oranges. Because Arrowhead scale-dust stuck to the leaves was suspected to be the allergen, tests of allergy were performed using an extract of the allergen prepared by Unger's method in our laboratory. Asthmatic attack was provoked 90 minutes after inhalation of the extract of cocoon. Histamine was not released, but leukotriene D4 production was induced by the addition of the extract to whole blood. Basophils were activated by addition of anti-IgG anti-sera as well as the extract. These data indicate that LT released from target cells in response to the worm elements, as well as histamine, is important as a chemical mediator.

Ca(2+)-activated K+ channel is present in guinea-pig but lacking in rat hepatocytes.

The mechanisms of norepinephrine-induced membrane responses in isolated hepatocytes from guinea-pigs and rats were compared using the suction-pipette, patch-clamp method, and intracellular Ca2+ concentration ([Ca2+]i) was measured using the Ca2+ fluorescent dye, Quin 2. The resting membrane potentials of isolated guinea-pig hepatocytes were -50 +/- 1 mV (mean +/- SD; n = 38), which is similar to that previously reported in rat hepatocytes by Sawanobori et al. (J Cell Physiol 139: 580-585, 1989). In guinea-pig hepatocytes, norepinephrine (6 microM) caused a membrane hyperpolarization, and norepinephrine (6 microM) or Ca(2+)-ionophore (A23187) (0.4 microM) caused a corresponding outward current. The sensitive current produced by norepinephrine and Ca(2+)-ionophore reversed its polarity at -74 +/- 9 mV (n = 7). The single channel recorded by cell-attached patch and inside-out patch had mean conductance of around 20 + 1 pS and was activated by 1 microM [Ca2+]i. On the other hand, neither norepinephrine (6-20 microM) nor Ca(2+)-ionophore (A 23187) (0.4 microM) caused any change in membrane potential and current in rat hepatocytes, whereas norepinephrine increased [Ca2+]i both in rat and guinea-pig hepatocytes to a similar degree. In the single-channel recording, we recorded single channels that had a mean conductance of 109.8 +/- 17.7 pS different from around 20 pS in guinea-pig. In inside-out patches, increased Ca2+ concentration from 10(-6) to 10(-3) M at the intracellular face of the membrane did not modify the single channel of rat hepatocytes. These results indicate that increased [Ca2+]i activates this channel in guinea-pigs, but that the channel activated by increased [Ca2+]i is lacking in rat hepatocytes membrane. Therefore, different mechanism operates in different species of liver cells to keep the constant state.

A family of familial hypercholesterolemia with cerebral infarction and without coronary heart disease. An unusual case with corneal opacity, polyneuropathy and carpal tunnel syndrome in the family: therapy with probucol and tocopherol nicotinate.

A study is presented of a 48-year-old female patient and her three siblings with familial hypercholesterolemia. The family members had episodes of cerebral infarction and apparently had atherosclerosis of the internal carotid artery, but no coronary heart disease due to their almost normal level of cholesterol. The laboratory studies of the family members revealed the elevations of serum lipid peroxides, serum lipoprotein(a), leukotriene C4 in blood, the thromboxane B2/6-keto-prostaglandin F1 alpha ratio in plasma and serum hydroxyl radical. Therefore, it is suspected that these factors accelerating atherosclerotic process caused the cerebral infarction. The patient demonstrated corneal opacities, palpebral xanthomas, thickened Achilles tendons, polyneuropathy and the carpal tunnel syndrome. Laboratory studies revealed an elevation in the OKT4/8 ratio, monocyte dysfunction with respect to phagocytosis and chemotaxis, and the presence of the 46XX/45XO mosaic chromosome. Lipid deposits were observed in the Achilles tendon, the transverse carpal ligament, the Schwann's cells and axons of the sural nerve, and in the keratocytes and stroma of the cornea. Following the administration of tocopherol nicotinate and probucol, the patient's serum lipid peroxide normalized and there was improvement in her palpebral xanthomas, thickening of the Achilles tendons and polyneuropathy. We conclude that the lipid deposits in this patient were due to the abnormal oxidative metabolism of low-density lipoprotein and a disturbance of the scavenger pathway due to the monocyte dysfunction.

[In vitro activities of newly developed quinolones, fleroxacin, lomefloxacin and sparfloxacin against Mycobacterium tuberculosis].

In vitro antituberculous activities of three newly developed quinolones, fleroxacin (FLRX, AM-833), lomefloxacin (LFLX) and sparfloxacin (SPFX, AT-4140) were evaluated in comparison to that of ofloxacin (OFLX) using M. tuberculosis strains isolated from patients and the Ogawa egg medium. SPFX was apparently more active than OFLX, but both FLRX and LFLX were less active. SPFX inhibited completely the growth of all 20 strains of M. tuberculosis isolated from patients who were not previously treated with OFLX in a concentration of 1.25 micrograms/ml. However, this agent inhibited the growth of only 4 strains (28.6%) of 14 OFLX-resistant M. tuberculosis in a concentration of 1.25 micrograms/ml, suggesting a partial cross-resistance between SPFX and OFLX.

Organic anion transport study in mutant rats with autosomal recessive conjugated hyperbilirubinemia.

The EHBR is a mutant rat strain with congenital conjugated hyperbilirubinemia bred from a Sprague-Dawley rat. Transport of conjugated bilirubin, indocyanine green, and tetrabromosulfophtalein from liver to bile is severely impaired in these rats. Serum bilirubin amounts to 6.0 +/- 0.05 mg/dl (n = 4) in adult rats, with 97% conjugates. The bile flow is reduced to about 65% of the control group, whereas total bile acid in 10-min bile samples is similar. Liver histology of 10 week-old rats revealed neither intracellular pigmentation nor architectural abnormalities.

Degradation and metabolism of indocyanine green: high-pressure liquid chromatographic analysis.

Degradation of indocyanine green solution by exposure to light was studied by high-pressure liquid chromatography. Indocyanine green in an aqueous medium exposed to light changed rapidly into an unknown product. The plasma clearance rate and the biliary excretion rate of the unknown product were much slower than those of indocyanine green. Spectrophotometric scan revealed that the unknown product had almost the same absorption spectrum as indocyanine green. Therefore, if degraded indocyanine green solution were to be used in a liver function test, the clearance of indocyanine green assayed by spectrophotometry would apparently be much lower than that of undegraded indocyanine green. According to fast atom bombardment mass spectrometry, the molecular weight of the unknown product was 723, whereas that of indocyanine green was 775. The analysis of rat bile after injection of indocyanine green by high-pressure liquid chromatography revealed that about 1% of the administered indocyanine green was metabolized in the rat liver.