Effect of prothrombin complex concentrate on INR and blood coagulation system in emergency patients treated with warfarin overdose.

We investigated the effect of prothrombin complex concentrate (PCC) on the international normalized ratio (INR) and blood coagulation system in two emergent patients treated with warfarin for secondary prevention of cardioembolic stroke due to nonvalvular atrial fibrillation. An 80-year-old woman developed massive subcutaneous hemorrhage and swelling on her right upper extremity with weak pulsation of the right radial artery and had an INR above 10. An 83-year-old man had pleural effusion with an INR value of 6.69 and pleural puncture was immediately required. We administered 500 IU of PCC to the two patients (17.2 IU/kg and 12.5 IU/kg) with 10 mg of vitamin K. The INR decreased to 1.12 and 1.85, respectively, with an increase of plasma levels of protein C and coagulant factors IIa, VIIa, IXa, and Xa 10 min after administration. The plasma levels of the thrombin-antithrombin III complex increased (from 4.0 to 12.0 micro g/l and from 0.5 to 28.9 micro g/l, respectively, normal value <3.0), but prothrombin fragment 1+2 increased minimally 10 min after administration (from 0.4 to 1.1 nmol/ml and from 0.4 to 0.7 nmol/ml, respectively, normal value 0.4-1.4 nmol/ml). Plasma levels of D-dimer remained unchanged. The massive subcutaneous hemorrhage in the former patient improved in 14 days. Anticoagulation was restarted in the latter patient after 14 days of PCC administration. There were no embolic episodes during the month after PCC administration. In conclusion, a small amount of PCC may be effective in immediately correcting increased INR levels with increased plasma levels of protein C and coagulant factors IIa, VIIa, IXa, and Xa and may partially activate the coagulation system without any effects on plasma levels of D-dimer.

[Radiofrequency catheter ablation therapy in elderly patients with supraventricular tachycardia].

138 patients with Wolf-Parkinson-White (WPW) syndrome (n = 96), atrioventricular nodal reentrant tachycardia (AVNRT; n = 27) and the other supraventricular tachycardia (n = 15), were divided into two groups, a control group (less than 65 years old; n = 108) and an elderly group (more than 66 years old; n = 30). We then estimated the success rate and safety of radiofrequency ablation for supraventricular tachycardia in elderly patients. For WPW syndrome, there were 76 (97%) successes and 9 (13%) recurrences in the control group (n = 78). In the elderly group of WPW patients, the number of successes was 18 (100%) and the number of recurrences one (63%). In 27 patients with AVNRT, the number of successes was 26 (96%) and there were no recurrences. In 15 patients with some other supraventricular tachycardia, there were 11 patients (73%) successes and one recurrence (11%). Major complications consisted of cardiac tamponade in 2 patients, dissecting aneurysm in one patient and cerebral embolism in one patients. All major complications occurred in patients with WPW syndrome. The cause of the complications, except the cerebral embolism was manipulation of the electrical or ablation catheter. Three of four patients with major complications belonged to the control group. It is possible that radiofrequency catheter ablation for supraventricular tachycardia in elderly patients is safe and highly effective. However, it is still invasive therapy. Ablation on a left accessory pathway by the transaortic valve approach especially needs meticulous care.

Epitope mapping of a monoclonal antibody to human glutathione transferase P1-1 the binding of which is inhibited by glutathione.

Although the three-dimensional structure of human glutathione transferase (GST) P1-1 crystallized with a GSH analogue has been reported, its structure in the non-complexed form has not been determined. Four monoclonal antibodies to GST P1-1 were produced to facilitate structural analysis. Of these, one, clone d-1 of IgG2a isotype, dose-dependently inhibited the activity of GST P1-1 but did not affect the activities of either GST A1-1 or M1-1. On immunoblotting, the antibody reacted strongly with GST P1-1 and weakly with rat GST-P and mouse GST-II, indicating cross-reactivity with Pi-class forms but preferential reactivity with GST P1-1. When GST P1-1 and the antibody were incubated in the presence of 60 microM GSH, no inhibition of activity was found, whereas 1-chloro-2,4-dinitrobenzene had no effect at concentrations up to 10 microM. The binding of GST P1-1 to antibody adsorbed to Protein A-Sepharose was also prevented by both 0.1 mM GSH and N-ethylmaleimide treatment. Trypsin digests of GST P1-1 were resolved by HPLC and a peptide that reacted with the antibody was detected by absorption experiments. N-Terminal amino acid sequencing revealed the peptide to be in the C-terminal portion of the enzyme, stretching from amino acid residues 198 to 208. A synthetic peptide of this sequence also absorbed the antibody. These results suggest that both GSH bound to the active site and N-ethylmaleimide bound to the cysteine residue repress antibody binding to the C-terminal region. Thus this antibody may be useful for examining the steric configuration of the C-terminal and other regions of GST P1-1 in the absence of GSH.

[Factor analysis of bone mineral dynamics in patients with renal osteodystrophy].

Remarkably high accumulation of Tc-99m-MDP is seen in the skull on bone scintigraphy of patients with renal osteodystrophy (ROD), especially with secondary hyperparathyroidism. For the quantitative evaluation, the Factor Analysis (FA) was used for the early phase of bone scan. Tc-99m-MDP (14.8 MBq/body weight kg) was injected as a bolus through the medial antecubital vein. Dynamic acquisition of 40 x 30 sec frames were performed in a 64 x 64 matrix. For pre-processing, nine points smoothing was carried out, and then the region of interest was set on the frontal image of the head for the FA. The FA was performed with an 8 x 8 sampling corresponding to 64 dixels from 4,096 dixels. Bone factor was clearly extracted by the FA. Then, two original parameters were calculated. One is the bone radionuclide uptake counts (Bu) which is the product of the total radionuclide counts of skull and the contribution ratio, the other is the uptake ratio (Br) derived by the time activity curve (physiological component) of the FA. These parameters of ROD were significantly different compared to those of controls. The FA seems to be useful in quantitative evaluation of bone mineral dynamics.

[Studies on the clinical and bacteriological effects of ofloxacin on the typhoid patients and carriers].

Fundamental and clinical studies of OFLX were performed against the patients with typhoid fever and typhoid carriers. 1) Clinical and bacteriological effects: Eight patients with typhoid fever and 3 typhoid carriers were treated with OFLX. Daily doses of the agent were 900 mg in 5 adult patients, 600 mg in a child patient and 3 adult carriers. In one case of the remaining 2 adult patients, daily doses of the agent changed from 800 mg to 1200 mg and from 900 mg to 1200 mg in the other one. The duration of the treatment was 9, 14 or 21 days. Clinical efficacies of OFLX against the patients proved 4 cases were "excellent", 3 cases were "good" and one case was "poor". The eradication of Salmonella typhi recognized in all cases containing 3 carriers with the exception of the "poor" case. Adverse reactions were observed transiently in 3 patients, such a slight decrease of RBC count, decrease of granulocyte count and elevation of GPT value respectively. 2) Antimicrobial activity: MICs of OFLX against 40 strains of S. typhi were 0.05 micrograms/ml and 0.1 micrograms/ml. The MICs of NFLX, CPFX and T-3262 were almost the same as that of OFLX, and those of ENX, NY-198 and NA were higher than that of OFLX. The peaks of MIC of CP and ABPC, first choice drug against typhoid fever, were 1.56 micrograms/ml and 0.38 micrograms/ml respectively. 3) Serum concentration; Serum concentrations of OFLX were serially measured on 5 patients through the day. The concentrations of the drug were distributed from 0.82 micrograms/ml to 6.34 micrograms/ml at 6.30 a.m. and from 2.52 micrograms/ml to 11.2 micrograms/ml at 9:00 p.m. Those of the day time showed considerable individual differences.

Occurrence and ultrastructural localization of duck hepatitis B virus in the liver of ducks after experimental infection.

A sequential study was performed to investigate the occurrence and localization of duck hepatitis B virus in the liver of domestic ducks utilizing the indirect immunoperoxidase method and electron microscopy. Seventeen ducklings were injected intravenously with duck hepatitis B virus-positive serum within 24 hr after hatching and were subsequently sacrificed on the 2nd, 3rd, 4th, 5th, 27th and 44th day after injection. Nine ducklings were not injected and were used as a negative control. Duck hepatitis B virus DNA by spot hybridization using a [3P]-labeled probe occurred in trace amounts on the 2nd day and in large amounts on the 4th day after inoculation. Immunoreactivity for DHBV was seen in the hepatocytes, sporadically on the 2nd day and diffusely on the 4th day, and also in the biliary epithelial cells on the 27th day. Both kinds of cells revealed staining in the cytoplasm but not in the nucleus. Virus particles were recognized by electron microscopy in the hepatocytes beginning on the 4th day. The hepatocytes had many incomplete virus particles, 40 to 61 nm in diameter, and a few complete virus particles, 40 nm in diameter, in the cisternae of the rough and smooth endoplasmic reticula. Such particles and the endoplasmic reticulum showed reaction products for duck hepatitis B virus by immunoelectron microscopy. There were clusters of core particles, 27 nm in diameter, in the hyaloplasm around peroxisomes where an assembly of cores appeared to occur. No conspicuous virus particles were recognized in the biliary epithelial cells. The similarities and differences in virus localization between duck hepatitis B virus and hepatitis B virus are discussed.

Production of antibody associated with non-A, non-B hepatitis in a chimpanzee lymphoblastoid cell line established by in vitro transformation with Epstein-Barr virus.

A continuous cell line of chimpanzee lymphocytes producing an antibody specifically associated with non-A, non-B hepatitis (NANB) was established. Peripheral blood lymphocytes of a chimpanzee convalescent from experimental infection with NANB hepatitis were transformed in vitro by Epstein-Barr virus infection into lymphoblastoid cell lines. Supernatants of the cell cultures were screened by immunofluorescence for antibody activity against the liver tissue of a chimpanzee with NANB hepatitis. Nineteen of the 1402 cultures were found to be positive for the activity. Ten of these 19 gave cytoplasmic reactions and the remaining 9 gave nuclear reactions in hepatocytes. One culture (48-1) stably producing the antibody was further characterized. The antibody produced in 48-1 was IgM and gave granular cytoplasmic reactions in hepatocytes. Cloning of 48-1 was performed by the soft agar method and cloned cell lines stably producing the antibody were obtained. The 48-1 antibody reacted with liver biopsy specimens from 12 chimpanzees obtained during the acute or chronic phase of hepatitis caused by five different NANB strains, but not with biopsy specimens from chimpanzees with hepatitis A or B or from normal chimpanzees. In addition, examinations of serial liver biopsy specimens obtained from 2 chimpanzees experimentally infected with NANB hepatitis demonstrated that the antibody reacted with the biopsies obtained during the preacute, acute, and chronic hepatitis, but not with those obtained before inoculation, early incubation period, or during convalescence. The present results indicate the specific association of the antibody with NANB hepatitis. Immunoelectron microscopy revealed that the antibody reacted with the microtubular aggregates identical to those previously described in a patient and chimpanzees with NANB hepatitis.

[Antitumor activity of cis-dichlorodiammineplatinum(II) and its effect on cell cycle progression].

Antitumor activity of cis-dichlorodiammineplatinum(II) (cisplatin) on various mouse transplantable tumors was investigated. Cisplatin was active against a wide variety of the following tumor systems: L1210 leukemia, P388 leukemia, B16 melanoma, colon tumor 38, Ehrlich ascites and solid carcinoma, WHT squamous cell carcinoma, and human stomach cancer G/S heterotransplanted in nude mice. From the comparison of growth inhibitory effect by cisplatin with various other antitumor agents in cultured Ehrlich ascites carcinoma cells, cisplatin was found to be mainly a concentration depending drug, but also time depending, so that it was identified as a type Ib class drug proposed by Shimoyama. Effect of cisplatin on the cell cycle progression of Ehrlich ascites carcinoma cells in mice was studied by flow cytometry of DNA. At an early stage after administration of cisplatin, cell cycle progression was delayed in S phase and blocked in G2 phase. With the elapse of time block in G1 phase or G1-S boundary was observed and the cell population, partially synchronized in G1 phase or G1-S boundary, progressed slowly through S phase to be blocked in G2 phase finally.