[Treatment of superior sulcus tumor].

We reported on 34 patients with superior sulcus non-small-cell lung cancer and clinical outcome. It is necessary to select the most appropriate approach from preoperative examination and the degree of infiltration at the chest wall. Recently, various approach and preoperative chemoradiotherapy followed by surgical resection is effective for the treatment of superior sulcus tumor (SST), we should keep challenging for radical resection in mind.

Thermosensitive polymer-modified liposomes that release contents around physiological temperature.

To obtain temperature-sensitive liposomes which release their contents around the physiological temperature, we designed dioleoylphosphatidylethanolamine liposomes modified with copolymers of N-isopropylacrylamide and acryloylpyrrolidine. Copolymers of acryloylpyrrolidine and N-isopropylacrylamide, which exhibit a lower critical solution temperature around the physiological temperature, were prepared by free radical copolymerization using azobis(isobutyronitrile) as the initiator. The copolymers with anchors to the liposome membrane were obtained by using N, N-didodecylacrylamide as an additional comonomer. The copolymer having the anchor group at the terminal of the polymer chain was also synthesized by copolymerization of these monomers in the presence of 2-aminoethanethiol and subsequent conjugation of N, N-didodecyl succinamic acid to the terminal amino group of the copolymer. Calcein-loaded dioleoylphosphatidylethanolamine liposomes modified with these copolymers were prepared and release of the contents from these liposomes was investigated. It was found that the release from these copolymer-modified liposomes was promoted around and above the lower critical temperature of the copolymer. Also, the liposomes modified with the terminal anchor-type copolymer released the contents more drastically responding to a small temperature change than the liposomes modified with random copolymers containing N,N-didodecylacrylamide units as the anchor.

Developmental changes in energy substrate use by the heart.

Marked changes in intermediary metabolism occur during development of the heart. In the fetus, the heart utilises lactate and glucose as its main energy substrates, while in the adult, fatty acids are the main energy substrate. The transition from carbohydrate to fatty acid metabolism is a complex process which involves maturation of mitochondrial processes and dramatic changes in circulating levels of fatty acids and lactate. In addition, developmental changes in the use of energy substrates also involve changes in the regulation of the enzymes involved in both carbohydrate and fatty acid utilisation. This paper reviews these changes in intermediary metabolism which occur during myocardial development. The metabolic differences that exist between immature and adult hearts may explain the observed differences in the ability of immature hearts to withstand hypoxaemia or ischaemia.

Cloning and nucleotide sequence of a cellulase gene, casA, from an alkalophilic Streptomyces strain.

A gene encoding an endo-type semi-alkaline cellulase was cloned from an alkalophilic Streptomyces strain in Streptomyces lividans, and its nucleotide sequence was determined. Downstream from the transcriptional start point, which was determined by high-resolution S1 mapping, an open reading frame of 388 amino acids (aa) was present. The N-terminal amino acid sequence of the mature enzyme determined by an Edman degradation procedure suggested that the cellulase had an extraordinarily long leader sequence of about 70 aa. Comparison with the leader sequences of endoglucosidase H from Streptomyces plicatus and the cellulase from Cellulomonas fimi suggested that the semi-alkaline cellulase was processed in two steps during maturation.

Temperature-dependent interaction of thermo-sensitive polymer-modified liposomes with CV1 cells.

Egg yolk phosphatidylcholine liposomes modified with a copolymer of N-acryloylpyrrolidine and N-isopropylacrylamide having a lower critical solution temperature at ca. 40 degrees C were prepared and an effect of temperature on their interaction with CV1 cells was investigated. The unmodified liposomes were taken up by the cells approximately to the same extent after 3 h incubation at 37 and 42 degrees C. In contrast, uptake of the polymer-modified liposomes by CV1 cells decreased slightly at 37 degrees C but increased greatly at 42 degrees C, compared to the unmodified liposomes. Proliferation of the cells was partly prohibited by the incubation with the unmodified liposomes encapsulating methotrexate at 37 and 42 degrees C. The treatment with the polymer-modified liposomes containing methotrexate at 37 degrees C hardly effected the cell growth. However, the treatment at 42 degrees C inhibited the cell growth completely. It is considered that the highly hydrated polymer chains attached to the liposome surface suppressed the liposome-cell interaction below the lower critical solution temperature of the polymer but the dehydrated polymer chains enhanced the interaction above this temperature. Because interaction of the polymer-modified liposomes with cells can be controlled by the ambient temperature, these liposomes may have potential usefulness as efficient site-specific drug delivery systems.

Radioactive tracers in congenital heart disease.

Radionuclide angiocardiography is a useful method in the evaluation of patients with congenital heart disease, safely and nontraumatically. Physiologic variables such as transit times, cardiac output, left ventricular ejection fraction, stroke volume, end-diastolic volume can be measured accurately with this technique. An important application of radionuclide angiocardiography in children with congenital heart disease is in the detection, localization and quantification (pulmonary to systemic flow ratio) of intracardiac shunts and shunts between the great vessels. This technique has been found useful in the evaluation of the newborn infant with cyanosis, the patient with a cardiac murmur and the patient who has had cardiovascular surgery. Newer mobile gamma camera-computer systems permit the performance of radionuclide angiocardiography in several ill premature and newborn infants and patients during the early postoperative period. Specially designed magnifying collimators and the development of ultrashort-lived radionuclides should result in an overall improvement in the diagnostic capabilities of this technique and in a further reduction in the radiation dose.

Surgical repair of double-chambered right ventricle with or without ventriculotomy.

Double-chambered right ventricle is an uncommon congenital anomaly consisting of obstruction of the trabecular zone of the ventricle by an elevated, hypertrophied moderator band, with or without additional muscle bundles(s). During an 18 month period (August 1984 to February 1986) 12 patients underwent repair of double-chambered right ventricle. Their ages ranged from 13 months to 12 years (mean 4.4 years). Associated lesions were ventricular septal defect in 11 patients (three closed spontaneously), atrial septal defect in two, left superior vena cava in three, aortic valve prolapse in one, and mild subaortic stenosis in two. Right ventricular outflow tract gradients were from 30 to 165 mm Hg (mean 77.2 +/- 37.3 mm Hg). The initial six patients (Group 1) were treated in the "classical" manner with a right ventriculotomy for muscle resection and ventricular septal defect closure (4/6). All patients survived. Five of the six required pharmacologic support for signs of mild to moderate right ventricular failure. The right ventricular outflow tract gradient postoperatively was 0 mm Hg in five patients and 25 mm Hg in one patient. Oxygen saturations were checked in the right atrium and pulmonary artery in all six patients, and one patient had a step-up of more than 10%. In the next six patients (Group 2) a previously unreported right atrial-pulmonary artery approach was used to avoid a ventriculotomy. The right ventricular outflow tract gradient was 0 mm Hg in five patients and 5 mm Hg in the sixth. A ventricular septal defect was closed in four of the six patients. There were no deaths, and no patient required inotropic support. Follow-up has been from 2 to 20 months. There have been no late deaths, and all patients are asymptomatic. One patient in Group 1 still has signs of a residual ventricular septal defect at 1 year. No patient has echocardiographic evidence of right ventricular outflow tract obstruction. All patients have remained in sinus rhythm with right bundle branch block. No patient has been recatheterized. Repair of double-chambered right ventricle is easily performed without a ventriculotomy. The atrial exposure of the ventricular septal defect is excellent. The muscle bundle(s) can be visualized easily through an initial transverse pulmonary arteriotomy with completion of resection through the tricuspid valve.

Subaortic stenosis in childhood: frequency of associated anomalies and surgical options.

In a 4-year period from July 1984 to July 1988, 21 children required surgical treatment for subaortic stenosis. Age at operation ranged from 12 months to 17 years (mean, 7.3 +/- 4.8 years). Associated anomalies were common and occurred in 13 patients (62%), including aortic valve insufficiency (eight), and stenosis (two), mitral valve stenosis (two) and insufficiency (one), and pulmonary valve stenosis (two) and absent pulmonary valve syndrome (one). A ventricular septal defect was present in five patients, and coarctation and patent ductus arteriosus occurred in three patients each. One patient had a complete atrioventricular septal defect, and one had an aortopulmonary window. Six patients had 15 previous procedures. The left ventricular to aortic gradient ranged from 20 to 170 mm Hg (mean, 59 +/- 43 mm Hg). The operation consisted of a transaortic resection (17), transventricular septal defect resection (two), a modified Konno procedure or septoplasty with preservation of the aortic valve (one), and a combined aortoventriculoplasty with homograft reconstruction of the ascending aorta and coronary reimplantation (one). There were no early deaths. One patient required pacemaker insertion for heart block. Follow-up ranged from 1 to 48 months (mean, 26.4 +/- 11.6 months). There was one late death 2 years after operation in the child with the aortopulmonary window who had Heath Edwards Type IV changes on a lung biopsy specimen at the original operation. One patient required reoperation for a residual gradient of 34 mm Hg 1 year after resection. Three patients still have aortic insufficiency. No cases of iatrogenic aortic valve injury occurred. Patients with subaortic stenosis represent a heterogeneous group. Early resection and additional procedures can be performed with a low mortality rate and can eliminate aortic insufficiency in many cases.

Intracardiac repair of dextro-transposition of the great arteries in the newborn period. Early and late results.

Between November, 1978, and December, 1981, nine neonates with dextro-transposition of the great arteries (d-TGA) and intact ventricular septum underwent primary intracardiac repair (i.e., because of persistent hypoxemia following balloon atrial septostomy arterial Po2 between 17 and 27 mm Hg). Age and weight at operation ranged from 2 to 21 days (mean 6.7) and from 1,900 to 4,500 gm (mean 3,493), respectively. The Mustard procedure was used in the first three patients and the Senning technique in the last six. All operations were done with deep hypothermia and circulatory arrest lasting 43 to 88 minutes (mean 70). An associated patent ductus arteriosus was ligated in eight of the nine patients. There were no deaths. The patients have been followed up for an average of 23 months (range 1 to 36). All are asymptomatic and require no medication. In one patient, atrioventricular dissociation necessitated implantation of a pacemaker 2 weeks after operation. All others are in sinus rhythm with normal atrioventricular conduction. Hemodynamic and angiographic follow-up was obtained in seven patients 8 to 25 months postoperatively (average 14.5). Left ventricular pressure was less than 30 mm Hg and aortic oxygen saturation was greater than 90% in all patients. One of the patients who had the Mustard procedure had complete obstruction at the junction of the inferior vena cava and the lower limb of the baffle. No evidence of systemic venous obstruction was found in the patients who had the Senning procedure. Early primary intracardiac repair appears to be a satisfactory alternative in the treatment of neonates with persistent hypoxemia following balloon atrial septostomy.

Skeletal muscle metabolism in the chronic fatigue syndrome. In vivo assessment by 31P nuclear magnetic resonance spectroscopy.

BACKGROUND: Previous study of patients with chronic fatigue syndrome (CFS) has demonstrated a markedly reduced dynamic exercise capacity, not limited by cardiac performance and in the absence of clinical neuromuscular dysfunction, suggesting the possibility of a subclinical defect of skeletal muscle. METHODS: The in vivo metabolism of the gastrocnemius muscles of 22 CFS patients and 21 normal control subjects was compared during rest, graded dynamic exercise to exhaustion and recovery, using 31P nuclear magnetic resonance (NMR) spectroscopy to reflect minute-to-minute intracellular high-energy phosphate metabolism. RESULTS: Duration of exercise was markedly shorter in the CFS patients (8.1 +/- 2.8 min) compared with the normal subjects (11.3 +/- 4.3 min) (p = 0.005). There were large changes in phosphocreatine (PCr), inorganic phosphate (Pi), and pH from rest to clinical fatigue in all subjects, reflecting the high intensity of the exercise. The temporal metabolic patterns were qualitatively similar in the CFS patients and normal subjects. There were early and continuous changes in PCr and Pi that peaked at the point of fatigue and rapidly reversed after exercise. In contrast, pH was relatively static in early exercise, not declining noticeably until 50 percent of total exercise duration was achieved, and reaching a nadir at 2 min postexercise, before rapidly reversing. There were no differences in pH at rest (7.08 +/- 0.04 vs 7.10 +/- 0.04), exhaustion (6.85 +/- 0.17 vs 6.76 +/- 0.17) or early (6.64 +/- 0.25 vs 6.56 +/- 0.24) or late recovery (7.09 +/- 0.04 vs 7.10 +/- 0.05), CFS patients vs normal subjects, respectively (NS). Neither were there intergroup differences (NS) in PCr or Pi. Although, quantitatively, the changes in PCr, Pi, and pH were marked and similar in both groups from rest to exhaustion, the changes all occurred much more rapidly in the CFS patients. Moreover, adenosine triphosphate (ATP) was significantly (p = 0.007) less at exhaustion in the CFS group. CONCLUSIONS: Patients with CFS and normal control subjects have similar skeletal muscle metabolic patterns during dynamic exercise and reach similar clinical and metabolic end points. However, CFS patients reach exhaustion much more rapidly than normal subjects, at which point they also have relatively reduced intracellular concentrations of ATP. These data suggest a defect of oxidative metabolism with a resultant acceleration of glycolysis in the working skeletal muscles of CFS patients. This metabolic defect may contribute to the reduced physical endurance of CFS patients. Its etiology is unknown. Whether CFS patients' overwhelming tiredness at rest has a similar metabolic pathophysiology or etiology also remains unknown.

Regulation of natural killer cell-mediated swine endothelial cell lysis through genetic remodeling of a glycoantigen.

The effect of remodeling of a glycoantigen such as the alpha-Gal epitope, Galalpha1,3Galbeta1,4GlcNAc-R, by the introduction of glycosyltransferase genes on natural killer (NK) cell-mediated direct cytotoxicity was investigated using human peripheral blood mononuclear cells (PBMC) or an NK-like cell line, YT cells, as an effector, and swine endothelial cells (SEC) as a target. Several SEC transfectants were established by transfection with the genes for beta1,4-N-acetylglucosaminyltransferase III, alpha2, 3-sialyltransferase and alpha1,2-fucosyltransferase. These transfections led to dramatic reductions in both direct and indirect NK cell-mediated cytotoxicity, by 72-94% in the case of PBMC and 27-72% in that of YT cells, in addition to an effective reduction in xenoantigenicity, which is substantially caused by the alpha-Gal epitope, to human natural antibodies. The NK cell-mediated direct cytotoxicity was remarkably blocked by an anti-alpha-Gal epitope monoclonal antibody or GSI lectin which preferentially binds to the epitope. Furthermore, treatment of the parental cells with alpha-galactosidase resulted in a significant reduction in cytotoxicity. These results suggest that the alpha-Gal epitope is involved not only in hyperacute rejection and acute vascular rejection, but also in NK cell-mediated direct cytotoxicity. Thus, the genetic remodeling of the alpha-Gal epitope and probably other glycoantigens as well can be expected to represent a new approach for overcoming not only indirect but also direct immunity to xenografts.

Cloning and sequencing analysis of Trp1 gene of Flammulina velutipes.

The genomic TRP1 gene from basidiomycete Flammulina velutipes was cloned by complementation of yeast Saccharomyces cerevisiae trp1 mutation. Sequencing analysis revealed that the TRP1 gene encoded a single protein consisting of three catalytic functional domains; glutamine amidotransferase, indole-3-glycerol phosphate synthase ) and N-(5'-phosphoribosyl) anthranilate isomerase, in order of NH2-glutamine amidotransferase-indole-3-glycerol phosphate synthase N-(5'-phosphoribosyl) anthranilate isomerase-COOH. The coding sequence of the TRP1 gene was interrupted by a single intron of 48 bases, the position and flanking sequences of which were highly homologous to those of basidiomycete Phanerochaete chrysosporium trpC.

Basidiomycetous fungus Flammulina velutipes harbors two linear mitochondrial plasmids encoding DNA and RNA polymerases.

Basidiomycetous fungus Flammulina velutipes R15 strain had two linear plasmids in its mitochondria designated pFV1 and pFV2. They were double-stranded DNAs, whose sizes were 8.3 and 8.9 kb, respectively. Sequencing analysis of 7364 bases of the pFV1 and 6861 bases of the pFV2 revealed that the both plasmids had one set of two open reading frames (ORFs) each of that encoded putative DNA and RNA polymerases similar to those of mitochondrial plasmids in other filamentous fungi. In phylogenetic analysis of deduced amino acid sequences of the ORFs and counterparts of other filamentous fungi, the pFV2 was expectedly clustered with plasmids of basidiomycetous fungi. whereas the pFV1 with kalilo plasmid of ascomycetous fungus Neurospora intermedia.

Evaluation of TMN classification for lung carcinoma with ipsilateral intrapulmonary metastasis.

BACKGROUND: Staging for lung cancer based on the TNM classification is an important predictive factor for prognosis. Recently, lung cancer with ipsilateral intrapulmonary metastasis (PM) was reclassified according to the revision of the TNM classification. To evaluate the prognostic importance of the new staging system for PM, we analyzed the postoperative survival of patients with non-small cell lung carcinoma. METHODS: Of 1,002 consecutive patients who underwent operation for primary lung cancer between June 1984 and December 1996, we reviewed the medical record of 889 patients who underwent complete resection for non-small cell lung cancer. RESULTS: We considered 89 patients (10.0%) to have synchronous ipsilateral PM. After reclassification to the former staging system revised in 1992, 5 patients were classified as stage I, 29 as stage IIIA, 48 as stage IIIB, and 7 as stage IV. In the new staging system revised in 1997, 48 patients were recategorized as stage IIIB, and 41 as stage IV. The 5-year survival of patients without PM (49.5%) was significantly better than that of patients with PM in primary-tumor lobe (29.6%, p = 0.002) or in nonprimary-tumor ipsilateral lobe (23.4%, p = 0.0002). Although the survival of patients with stage IV cancer without PM was significantly worse than that of patients with the new (1997) stage IV cancer with PM (p = 0.02), it was similar to that of patients with the former (1992) stage IV cancer with PM. The survival of PM patients with N0 or N1 disease was significantly better than that of PM patients with N2 or N3 disease (p = 0.001). Furthermore, in patients with the new (1997) stage IIIB cancer, the survival of N0 disease was better than that of N2 disease (p = 0.007). CONCLUSIONS: Inasmuch as the prognosis of non-small cell carcinoma in patients with PM strongly correlated with N factor rather than PM factor, N factor should be reflected in a staging designation. We therefore consider the new TNM classification for PM reclassified in 1997 to be less acceptable for surgical-pathologic staging than the revision in 1992.

Elevated serum levels of neuron specific enolase in lymphoid leukaemia.

Using an enzyme-linked immunosorbent assay (ELISA), we measured the levels of neuron-specific enolase (NSE) in the sera of 100 normal controls, 31 patients with adult T cell leukaemia (ATL), 13 patients with acute lymphoblastic leukaemia (ALL), 19 patients with acute non-lymphoblastic leukaemia (ANLL) and 17 patients with human T lymphotropic type I (HTLV-I) associated myelopathy (HAM). Levels higher than 5 ng/ml were considered as positive for NSE, since this value was almost 2 standard deviations (S.D) above the mean of normal controls. The rates of positive NSE levels were 55 percent in patients with ATL, 54 percent in patients with ALL, 11 percent in patients with ANLL and 0 percent in patients with HAM. High NSE levels fell to near normal levels in patients with ATL and ALL responding to cytotoxic therapy. It is concluded from these results that NSE may be a useful tumour marker for lymphoid malignancies.

Pulmonary vascular obstructive disease.

The history, clinical course, pathophysiology, pathology, diagnosis, and therapy of pulmonary vascular obstructive disease are presented. An explanation is provided for the terms Eisenmenger syndrome, Eisenmenger complex, and pulmonary vascular disease. The complications frequently seen in the natural history of the condition are presented, as are the therapeutic options. Despite intensive investigation of new therapeutic products, the most promising treatment for established pulmonary vascular obstructive disease remains heart-lung operation or single-lung transplant with repair of cardiac defects. Even with this option, the long-term outlook for this progressive condition remains dismal.

Aortic atresia with a solitary and indeterminate ventricle.

A male infant having aortic atresia and double inlet to a solitary and indeterminate ventricle, presented a clinical picture of "hypoplastic left heart syndrome". Haemodynamic consequences are described and comparison made to other types of aortic atresia. Cross-sectional echocardiography was found to be reliable in determining both atrioventricular and ventriculo-arterial connections in the presence of aortic atresia and a solitary ventricle.

Feedback regulation of hand grasp opening and contact force during stimulation of paralyzed muscle.

A fixed-parameter, discrete-time, first-order, feedback control system is described for regulating grasp during electrical stimulation of paralyzed muscles of the hand. The stiffness of the grasp (relationship between grasp force and grasp opening) is kept constant by linearly combining force and position feedback signals. Thus, a single continuous command signal can control the size of the grasp opening prior to object acquisition and both grasp force and opening after contact. The controller achieves this change in controlled variables by scaling and summing the force and position feedback signals, rather than by a discrete switch in control strategy. Experimental tests of the control system in quadriplegic subjects show that control can be obtained over conditions ranging from unloaded position regulation to isometric force regulation, as well as in the transition between these conditions. The robustness of the control system was evaluated during force regulation with isometric loads. Step response rise time and overshoot were much more dependent on system gain than on the location of the controller zero. Responses with rise time less than two seconds and overshoot less than 30% were obtained over a gain range up to ten, indicating good robustness to muscle gain reductions such as might be caused by fatigue.

Open-loop control of the freely-swinging paralyzed leg.

An experimental model has been used to study issues that are relevant to the use of electrical stimulation to help paralyzed individuals walk. Modulated stimulation sequences for the quadriceps muscles were manually selected using an iterative trial-and-error procedure to cause the knee angle to follow a specific movement pattern (desired trajectory). Four paraplegic subjects were tested before and after an eight-week program in which the quadriceps were exercised daily with electrical stimulation. It was found that 12.6 +/- 2.9 iterations were required to approximate the desired trajectory. The average error of the final match between the actual and desired trajectories was 2.1 degrees +/- 0.7. Repeated responses were extremely consistent; the average difference between successive trials was less than 1 degree in 81 percent of the trials. When the stimulation sequence was repeated every 3 s for 50 cycles, however, there was a progressive degradation in the response, even in exercised legs, that demonstrated the limitations of open-loop control. Stimulus modulation envelopes for all four subjects were similar in shape (although varied in amplitude) indicating that the iterative process can be shortened by starting with an "average" modulation envelope. Stimulation sequences achieved accurate matches of the desired trajectory on subsequent days when adjusted by a simple gain factor. The relevance of these results to multichannel control of walking is discussed.

Micropropagation ofPanax notoginseng by somatic embryogenesis and RAPD analysis of regenerated plantlets.

Somatic embryogenesis was induced in callus tissues derived from young flower buds ofPanax notoginseng via callus within 18 weeks of culture. The mature somatic embryos were germinated on half-strength Murashige and Skoog's (MS) medium supplemented with gibberellic acid A3(GA) and 6-benzyladenine (BA). The most suitable medium for optimal root formation proved to be MS medium supplemented with 1-naphthaleneacetic acid (NAA). Total DNA was extracted from the leaves of the regenerated plantlets ofP. notoginseng. Analysis of random-amplified polymorphic DNA (RAPD) using 21 arbitrary oligonucleotide 10-mers, showed the genetic homogeneity ofP. notoginseng. The amplification products were monomorphic for all of the plantlets ofP. notoginseng regenerated by embryogenesis, suggesting that somatic embryogenesis can be used for clonal micropropagation of this plant.