[Patent foramen ovale presented hypoxemia with cardiac sarcoidosis].

A 53-year-old woman with a double, double, double (DDD) pacemaker due to complete atrioventricular block was admitted to our hospital with a diagnosis of congestive heart failure. At the time of admission, she was in a hypoxic state with cyanosis and clubbed finger. The ultrasonic cardiogram showed a severe degree of tricuspid valve regurgitation and a thin left ventricular septal wall. Transesophageal echocardiography revealed a patent foramen ovale (PFO) with continuous right to left shunt flow. She was diagnosed with cardiac sarcoidosis with hypoxemia caused by PFO. PFO closure and tricuspid valve annuloplasty (DeVega method) were performed. Following surgery, the patient's hypoxemia improved and the cyanosis disappeared. The patient was discharged 37 days after the operation.

Resuscitation of Vibrio cholerae O1 strain TSI-4 from a viable but nonculturable state by heat shock.

Vibrio cholerae strain TSI-4 was incubated in an M9 salt solution at 15 degrees C for more than 100 days. The plate counts showed no viable cells on day 30, but a broth culture from that day showed the growth of bacteria. However, after 35 days the bacteria entered the nonculturable state, based on the assessment of both the plate counts and broth culture. A portion of the culture was heated at 45 degrees C for 1 min in a water bath and subsequently plated onto a nutrient agar plate. More than 1000 colonies were recovered after this heat-shock treatment. The recovered cells showed the same chromosomal DNA pattern in the restriction map and the same outer membrane protein pattern in SDS-PAGE. Recovery of viable cells by heat-shock was achieved in cultures grown on M9 salt but not from cultures grown in phosphate-buffered saline. This suggests that the presence of NH4Cl in the M9 salt solution may support the growth of the bacteria in a low nutrient medium, while also playing an important role in resuscitation.

Characterisation of the external surfaces of Pseudomonas aeruginosa isolated from human blood and respiratory tract.

The surface structures of the cell envelopes of 16 clinical isolates of Pseudomonas aeruginosa were examined by electronmicroscopy with the new fixation technique of freeze-substitution. Two types of structures were observed among the organisms. In one group of strains, mostly isolated from blood, a dense fibrous layer c. 30 nm thick was found around the outer-membrane surface, whereas no such structure was observed in the other group of isolates, most of which were from sputum. Lipopolysaccharides extracted from the isolates with a dense fibrous layer were found by SDS-PAGE to have long O-polysaccharide chains, whereas strains without such a layer mostly had lipopolysaccharides that lacked high mol. wt. O-polysaccharide chains.

Seasonal development of Leptotrombidium pallidum (Acari: Trombiculidae) observed by experimental rearing in the natural environment.

Engorged larvae of Leptotrombidium pallidum Nagayo, Miyagawa, Mitamura & Tamiya, the vector mite of scrub typhus in Japan, were reared by feeding them with fresh eggs of the collembolan Sinella curviseta Brook while confined in small plastic containers under natural conditions in a copse. The larvae were collected from wild rodents (Apodemus speciosus) in autumn 1985 and spring 1986. Adults were kept alive for 2 yr or longer. The larvae obtained in autumn became dormant in the cold winter season, and growth recommenced in the spring. Thus, the development of mites collected in April became synchronized with that of larvae obtained in the autumn. Most larvae developed into protonymphs in May, deutonymphs in June, tritonymphs in July, and adults in August. The females laid eggs in two consecutive summers. Some larvae collected in autumn were kept in a refrigerator until the following summer. They developed into deutonymphs, tritonymphs, or adults and then became dormant in the winter. Development restarted the next spring and all became adult by summer, when the females laid eggs. Under experimental conditions, all larvae are hatched in the autumn, unlike the natural situation in which two peaks of larval occurrence on wild rodents are observed in autumn and spring.

Seasonal development of Leptotrombidium akamushi (Acari: Trombiculidae) under field temperatures.

Engorged larvae of Leptotrombidium akamushi (Brumpt), a vector of scrub typhus, were reared in small plastic containers placed on the ground and fed fresh eggs of the collembolan Sinella curviseta Brook. Engorged larvae obtained in October developed into deutonymphs through protonymphs approximately 1 mo before winter and became dormant in the cold winter season (approximately 3 mo). Most deutonymphs developed into tritonymphs in April and adults in May. Females began laying eggs in mid-June and the numbers of unfed larvae showed a peak in August. The mites reared from July rapidly developed into adults by August, and laid eggs in September. Larvae were most abundant in October, and adults became dormant in the winter. The same adults laid eggs from early May to late June and, upon hatching, the larval population peaked in early July of the 2nd summer. Most larvae died before the 2nd winter. Eggs hatched approximately 3 wk after oviposition and longevity of unfed larvae was 2 mo. Because of this very short incubation period, L. akamushi larvae occur in the summer, whereas L. pallidum Nagayo, Miyagawa, Mitamura & Tamiya, and L. scutellare Nagayo, Miyagawa, Mitamura, Tamiya & Tenjin occur in the autumn, although 3 species lay eggs from May to August.

Failed vertical transmission of Rickettsia tsutsugamushi (Rickettsiales: Rickettsiaceae) acquired from rickettsemic mice by Leptotrombidium pallidum (Acari: trombiculidae).

Larvae of Leptotrombidium pallidum (Nagayo, Miyagawa, Mitamura & Tamiya) from uninfected laboratory colonies were fed on mice infected with Rickettsia tsutsugamushi (Hayashi) Ogata. Infection of the chiggers with R. tsutsugamushi was determined by passage of chigger exudates into ddY mice. The passage method was modified so that an inoculum was considered to be positive when R. tsutsugamushi or anti-R. tsutsugamushi antibody, or both, were detected in mice up to the third blind passage. R. tsutsugamushi was detected in six of 18 larvae (33.3%) and in all developmental stages. In adults, five of 18 males and 10 of 46 females were infected with R. tsutsugamushi. In L. fuji (Kuwata, Berge & Philip), R. tsutsugamushi was not found in 57 engorged larvae fed on rickettsemic mice but was found in a very low percentage of deutonymphs and adults. Female L. pallidum from larvae fed on infected mice were paired individually, and F1 larvae were collected. Although eight females were found to be positive for R. tsutsugamushi, the rickettsia was not detected in 12 pools (249 larvae) of F1 larvae from these infected females. We concluded that uninfected mites became infected by feeding on rickettsemic mice at comparatively high rates depending on the species and transmitted this infection transstadially to succeeding life stages, but not vertically to larvae in the following F1 generation.

Stability of RNA chain elongation complexes formed with RNA polymerase and denatured DNA templates.

A random transcription system was used to analyze RNA chain-elongation complexes with RNA polymerases I and II from mouse ascites sarcoma cells. The molecular size of synthesized RNA was analyzed to characterize the elongation reaction by RNA polymerases. The RNA chain-elongation complex after initiation of RNA synthesis on denatured DNA by RNA polymerase II was not decomposed with heparin which dissociated the DNA-RNA polymerase complex, whereas the elongation complex on denatured DNA by RNA polymerase I was more susceptible to decomposition with heparin. RNA transcribed by RNA polymerase I on denatured DNA 30 min after the start of the reaction contained large RNA around 28S and small RNA. The small RNA could be formed by reinitiation and early termination of elongation. RNA transcribed by RNA polymerase II on denatured DNA was only large RNA around 28S. The elongation reaction by RNA polymerases I and II on denatured DNA was not inhibited by rifamycin AF/013, an inhibitor of initiation. These results suggest that RNA polymerase II could synthesize large transcripts without reinitiation and early termination of elongation and that RNA polymerase II formed more a stable elongation complex on denatured DNA than RNA polymerase I.

A comparative study of DNA-dependent RNA polymerases from rat ascites hepatoma cell nuclei and from rat liver nuclei.

DNA-dependent RNA polymerases (EC 2.7.7.6) were extracted and partially purified form the nuclei of rat ascites hepatoma cells (AH-130) induced by 4-dimethylaminoazobenzene. The patterns of RNA synthesis and the properties of these enzymes were compared with enzymes from the nuclei of rat liver. The specific activity of RNA polymerase in the homogenate from the nuclei of AH-130 cells was the same as normal rat liver nuclei. RNA polymerase was solubilized from the homogenate at high ionic strength and separated into two forms by DEAE-Sephadex column chromatography. Enzymatic characterization showed that these enzymes corresponded to RNA polymerase I and II. RNA polymerase I more effectively transcribed native DNA than denatured DNA at low salt concentration, but at high salt concentration RNA polymerase I effectively transcribed denatured DNA. RNA polymerase II more effectively transcribed denatured DNA. In AH-130 cells the activity of RNA polymerase I was 4 to 5 times higher than RNA polymerase II, and in rat liver the activity of RNA polymerase I was 1.5 to 2 times higher than RNA polymerase II. The activity of RNA polymerase I in AH-130 cells may have increased by induction.

Characterization of RNA polymerases from Rous sarcoma virus-induced mouse ascites sarcoma cells.

RNA polymerase was extracted from the Schmidt-Ruppin strain of Rous sarcoma virus (SR-RSV)-induced C3H/He mouse ascites sarcoma cells (SR-C3H). RNA polymerase was separated into RNA polymerases I and II by DEAE-Sephadex chromatography. RNA polymerase I was separated into Ia and Ib fractions by phospho-cellulose chromatography. In SR-C3H cells RNA polymerase Ib was the main component of RNA polymerase I. At 0.05--0.1 M ammonium sulphate RNA polymerase I transcribed native DNA most actively, and RNA polymerase II transcribed denatured DNA most actively. Partial digestion of DNA by DNAase I enhanced RNA synthesis by RNA polymerases I and II. At ionic strength over 0.2 M ammonium sulphate, the initiation reaction of RNA polymerases I and II was inhibited. The initiation complexes of RNA polymerases I and II with native DNA were more stable against high salt concentration than with denatured DNA.

RNA synthesis in permeable mouse ascites sarcoma cells.

A permeable cell system for studying RNA synthesis was established. Mouse ascites sarcoma cells were made permeable to nucleoside triphosphates and alpha-amanitin by treating with a hypotonic buffer. Separate determinations of endogenous RNA polymerase I, II and III activities in permeable cells were conducted using the different sensitivities of these enzymes to alpha-amanitin. The endogenous activity of RNA polymerase II under optimal conditions was one tenth of total RNA synthetic activity in isolated nuclei, and one third of that in permeable cells. The extremely low ratio of RNA polymerase II activity to total RNA synthetic activity in isolated nuclei was thought to be caused by increase of RNA polymerase I activity and decrease of RNA polymerase II activity. These and other results suggested that RNA synthesis in permeable cells reflects more precisely the in vivo state of RNA synthesis than thatin isolated nuclei. The permeable cell system will provide a useful method for studying the separate activities of RNA polymerases I, II and III in situ.

Effects of some polyamines, polyanions and antitumor drugs on replicative DNA synthesis and unscheduled DNA synthesis in vitro.

The effects of various compounds on replicative DNA synthesis in permeable mouse ascites sarcoma cells and on unscheduled DNA synthesis in permeable cells or in isolated rat liver nuclei were studied. Polyamines such as spermidine, putrescine and cadaverine inhibited replicative DNA synthesis. Unscheduled DNA synthesis was inhibited by spermidine and cadaverine, but slightly stimulated by putrescine at low concentrations. Aurintricarboxylic acid, a low molecular weight polyanion, inhibited both replicative DNA synthesis and unscheduled DNA synthesis. Replicative DNA synthesis was inhibited by heparin, a high molecular weight polyanion, whereas unscheduled DNA synthesis was stimulated at low heparin concentrations. Antitumor drugs such as daunomycin, neocarzinostatin and bleomycin inhibited replicative DNA synthesis. Unscheduled DNA synthesis was inhibited by daunomycin, slightly induced by neocarzinostatin and highly induced by bleomycin. The present system was thought to be useful for studying the separate effects of various drugs on either replicative DNA synthesis or unscheduled DNA synthesis in vitro.

Mange caused by Sarcoptes scabiei (Acari: Sarcoptidae) in wild raccoon dogs, Nyctereutes procyonoides, in Kanagawa Prefecture, Japan.

Parasitological and histopathological examinations were performed in 25 raccoon dogs (Nyctereutes procyonoides) obtained in Kanagawa Prefecture, Japan, all of which were found to be heavily infected with Sarcoptes scabiei. The mites detected on these raccoon dogs were morphologically indistinguishable from the human species, and no Demodex mites were detected. Histopathological examinations showed prominent hyperkeratosis and acanthosis with eczema, and numerous burrows containing mites were observed in the epidermis. The enzootic dermatitis of wild raccoon dogs in recent years was clearly demonstrated to be caused by S. scabiei in the present study.

[The effect of intraoperative high-dose tranexamic acid on blood loss after operation for acute aortic dissection].

The effect of high dose tranexamic acid on blood loss after operations for acute aortic dissection was evaluated. Twenty-eight patients undergoing emergent operations for acute aortic dissection were studied. There were two groups, group T with 13 patients (group T) who were given 7 g of tranexamic acid after induction of anesthesia and 3 g of it after CPB and group C with 15 patients who did not receive tranexamic acid. There was a tendency that group T had less bleeding during operation and after operation (559.6 +/- 865.8 ml in group T and 805.8 +/- 442.9 ml in group C, 1719.2 +/- 1008.7 ml in group T and 3547.7 +/- 4580.1 ml in group C, respectively), but there was no significant difference between two groups. The removal of drainage tubes after operation was significantly earlier in group T (5.0 +/- 2.3 post operative day in group T and 8.1 +/- 5.2 post operative day in group C; p < 0.05). FDP and D-dimer level as measures of fibrinolytic activity were elevated at pre- and postoperative period in both groups, but they tended to be lower in group T at postoperative period. One patient required reexploration because of excessive bleeding and no mediastinal infection was reported in group T, whereas 4 patients underwent reexploration and 2 patients developed mediastinitis in group C. There were 5 hospital death (33.3%) in group C and 2 (15.4%) in group T. High dose of tranexamic acid seems to control fibrinolytic activity, thereby reducing blood loss and requirements, which may contribute to lower morbidity and mortality in operations for acute aortic dissection.

[A case report of pulmonary valve blood cyst].

A six-year-old boy with pulmonary stenosis was hospitalized in order to examine the cause of pulmonary stenosis. The right-ventriculography showed a large shadow defect in the right ventricular outflow tract. But the echocardiography showed the cystic tumor, which was covered with thin wall and had echo-free space in it, in the right ventricular outflow tract. We made a diagnosis of pulmonary blood cyst. After cardiopulmonary bypass was established, the main pulmonary artery was incised longitudinally. There were two blood cysts at the left pulmonary cusp which were 1.5 x 1.5 cm and 0.7 x 0.5 cm in size. The cysts were resected and the pulmonary cusp was plicated. Histologically, the wall of the cysts was consisted thick valvular tissues. No pulmonary valve pressure gradient was detected with the echocardiography following the operation.

[A case of a left atrial primary malignant fibrous histiocytoma].

Primary malignant fibrous histiocytoma (MFH) of the heart is very rere. We report a case of a primary intracardiac MFH. A 50-year-old woman underwent an operation for congestive heart failure due to left atrial (LA) tumor. Cardiac echocardiogram showed the tumor attached to the septal wall and posterior wall of LA. We extendedly resected the tumor with septal and partially posterior wall and reconstructed there with pericardial substitute. Pathological diagnosis was MFH. Although she did well postoperatively, cardiac echocardiogram showed a recurrent tumor in the LA at 10 days after operation. She developed respiratory failure, and chest and abdominal CT demonstrated metabolism to the lung, liver and bladder at post operative 47 days. At 77 days after operation, she expired. The prognosis of MFH is poor despite surgery. We recommend extended surgery, if the LA tumor had wide base attached to the septal or posterior wall.

[A case of radical operation for right-sided infective endocarditis due to Pseudomonas aeruginosa concurrent with VSD in the elderly person].

The case is a 60-year-old male. He was seen at this department because of a slight fever of less than 37.5 degrees C from half a year earlier. Echocardiography revealed vegetations under the tricuspid valve and pulmonary valve, and on scintigraphy of the pulmonary blood flow he was diagnosed with multiple pulmonary infarction. Causative bacteria were not identified by blood culture on admission. As the intraoperative findings, vegetations were seen attached to the area from the septal cusp to the anterior cusp in the tricuspid valve. With the pulmonary valve, a part of the valvular cusp was perforated. After closing VSD with a patch, the vegetations attached to the valvular cusps were excised and the tricuspid valve was reconstructed. As for the pulmonary valve, valve replacement was performed because of the destructive degeneration of the valvular cusp. Pseudomonas aeruginosa was detected on culture of the tissue of the valvular cusp resected during operation. Post operative course was uneventful. The patient was discharged on the 21st postoperative day.

[CABG performed for LMT orifice stenosis 15 years after AVR in a case of aortitis syndrome].

The case is a 54-year-old female at age 39 years, she underwent aortic valve replacement (AVR) for aortic valve regurgitation (AR) due to aortitis syndrome at a certain hospital. At the 15th year after AVR she was diagnosed with unstable angina due to LMT orifice stenosis and underwent CABG at this hospital. A distal side of anastomosis was performed on the LAD branch seg 6, an artificial vascular patch was placed for the proximal side of anastomosis to prevent inflammation of the aortic wall from spreading to the proximal side of anastomosis. On the postoperative cardiac catheter examination, the graft was found to be patent well. In the case of CABG complicated with aortitis syndrome, stenosis and obstruction of the proximal side of anastomosis long time after surgery pose a problem. However, the method so as to perform a proximal anastomosis by placing an artificial vascular patch on the aortic wall is considered to be one of the useful means which can avoid such a problem.

[Surgical treatment of transposition of the great arteries with intact ventricular septum associated with left ventricular outflow obstruction].

Thirteen patients have undergone surgical treatment for transposition of the great arteries (TGA) with intact ventricular septum (IVS) associated with left ventricular outflow obstruction (LVOTO) in our institute. Ages at operation ranged from three months to six years (mean 9.2 +/- 4.7 months). Of these patients, seven had dynamic type LVOTO (group I), and six had organic LVOTO (group II). Preoperative left ventricular end-diastolic volume was significantly smaller in group II (78 +/- 13% of normal) than control group (135 +/- 53% of normal). Preoperative pressure gradient between the pulmonary artery and left ventricle was significantly greater in group II (55.8 +/- 3.2 mmHg) than group I (35.2 +/- 3.9 mmHg). In group I, II patients underwent two-stage ASO, the other 5 patients underwent Senning operation. The reason for the Senning operation were era before introduction of ASO in our institute (1983) or unsuccessful training of the left ventricle. In group II, all but one patient underwent Senning operation, the other underwent a successful Fontan operation with Damus anastomosis because of too small left ventricle (LVEDV: 49% of normal). Techniques to correct LVOTO at the definitive operation included ventriculotomy (n = 4) and pulmonary valvotomy (n = 2). One patients in group II underwent a Fontan operation with Damus anastomosis due to an underdeveloped left ventricle (LVEDV: 49% of normal). There was no early or late death. The postoperative pressure gradients disappeared or reduced to trivial levels in all patients. At present no LVOTO has developed in any of the patients.

[Pre and postoperative left ventricular volume and function in infants and children with total anomalous pulmonary venous return].

From January, 1989, to December, 1991, 36 consecutive patients with total anomalous pulmonary venous return (TAPVR) underwent intracardiac repair (ICR) in our institute. We studied the relationship between the size of atrial septal defect (ASD), age and preoperative left ventricular end-diastolic diameter (LVEDD) in 28 patients with regression analysis. There was no statistical relationship between the size of ASD, age and LVEDD. We measured LVEDD, RV to LV pressure ratio (RVP/LVP) and LV fractional shortening (LVFS) using two dimensional echocardiography in 15 patients, prior to and following ICR. LVEDD increased from 1.16 +/- 0.40 cm to 1.62 +/- 0.28 cm immediately following ICR, and then increased to 1.79 +/- 0.34 cm one month following ICR again. LVFS decreased from 38% to 35% immediately following ICR but still remained within normal range (normal range 28-45%). RVP/LVP decreased from 1.0 to 0.69 immediately following ICR, and to 0.39 one month following ICR. We concluded that the decrease of preoperative LV end-diastolic volume in TAPVR was not due to the decrease of LV preload but to anterior deviation of the ventricular septum caused by high RV pressure. There was significant increase of LV volume immediately following ICR, but LVFS maintained in normal range. This study suggested that small LV volume in patients with TAPVR did not influence the surgical results.

[Surgical treatment of total anomalous pulmonary venous connection Darling type Ib using pedicled right atrial flap].

A one-month-old baby with total anomalous pulmonary venous connection (TAPVC) type Ib underwent a total correction with a pedicled right atrial (RA) flap, which was made by incising the RA wall in a quadrangular configuration. The common pulmonary vein (PV) was cut back into the left atrium (LA). Then the RA flap was sutured along the limbus of PV recess and atrial septal defect (ASD) to create a new pulmonary venous channel. The defect in the RA wall was directly closed without any prosthetic patch. Absorbable sutures (# 6-0 PDS) were used throughout. Postoperative course was uneventful and echocardiogram showed widely opened PV channel draining into the LA. We think that this procedure could be applied in various types of total and partial anomalous pulmonary venous connection, avoiding pulmonary venous obstruction on the assumption that the RA flap should grow.