Problems in the current diagnostic standards of clinical N1 non-small cell lung cancer.

BACKGROUND: Although clinical N1 (cN1) non-small cell lung cancer (NSCLC) is considered to be locoregional, the postoperative outcome is disappointing, with a 5 year survival of less than 50%. One possible reason may be that cN1disease diagnosed by current standard imaging modalities often contains unexpected N2 disease. This study was conducted to evaluate the surgical and pathological results of patients with cN1 NSCLC. METHODS: Among 1782 patients with NSCLC who underwent intended curative resection from 1993 to 2003, 143 patients were identified as having cN1 disease and were enrolled in this study. The clinicopathological records and CT films of each patient were retrospectively reviewed to identify predictors for pN2-3 disease. RESULTS: The pathological nodal status was pN0 in 23% (n = 33), pN1 in 47% (n = 67) and pN2-3 in 30% (n = 43) of patients. Patients with pN2-3 showed a significantly worse 5 year survival rate of 38% compared with patients with pN0 (68%) and pN1 (60%) (p = 0.017 and 0.007, respectively). Multivariate analysis showed that adenocarcinoma histology was a significant predictor for pN2-3 disease (OR 3.312, 95% CI 1.439 to 7.784; p = 0.005). The presence of N1 node separate from the main tumour on CT scans tended to predict pN2-3 disease although this did not reach statistical significance (OR 2.103, 95% CI 0.955 to 4.693; p = 0.066). Pathological N2-3 disease was found in 53% of patients with adenocarcinoma with a separate N1 pattern and in only 12% of patients with non-adenocarcinoma with a continuous N1 pattern. CONCLUSIONS: The diagnosis of N1 status by contrast enhanced CT scans is unsatisfactory with a high rate of unexpected pN2 disease. To avoid infertile lung resection, patients with CT diagnosed N1 adenocarcinoma, especially with a separate N1 pattern on CT, should be considered for additional invasive node biopsy modalities, including mediastinoscopy.

Native and recombinant human hepatocyte growth factors are highly potent promoters of DNA synthesis in both human and rat hepatocytes.

Human hepatocyte growth factor (hHGF) has recently been expressed as a recombinant polypeptide from Chinese hampster ovary cell transfectants. Using a primary rat hepatocyte bioassay, we have tested the biological activity of recombinant hHGF and compared it with native hHGF. Dose-response curves were almost identical, with half-maximal stimulation of DNA synthesis at 1-2 ng/ml (equivalent to approximately 10 pM). S-phase labeling index was similarly enhanced and numerous mitotic cells were observed. Recombinant and native hHGF also stimulated DNA synthesis and S-phase labeling index in primary adult human hepatocytes. Human cells were more responsive than rat hepatocytes, with recombinant hHGF slightly more potent than native hHGF (half-maximal stimulation 0.3 and 0.6 ng/ml, respectively). Since HGF levels rise in patients with fulminant hepatic failure and in animals after partial hepatectomy or administration of hepatotoxins, situations where liver regeneration occurs, HGF is suggested to play a key role in regulation of hepatic growth. The high potency of the factor on human hepatocytes reinforces its candidacy as a critical mitogen in human liver growth. The availability of a recombinant hHGF opens the way for in vivo experimental studies and to the possibility of using hHGF as a clinical therapeutic agent, either alone or in combination with other factors.

Human intrahepatic biliary epithelial cells proliferate in vitro in response to human hepatocyte growth factor.

In previous studies, intrahepatic human biliary epithelial cells (BEC) were isolated in high purity. However, these cells demonstrated only limited growth responses. Here we report that human BEC proliferate in response to human hepatocyte growth factor (hHGF), retain BEC-specific phenotype, and can be serially passaged. BEC showed dose-dependent growth in response to 0.01-100 ng/ml hHGF. The maximum S-phase labeling index reached 40% with half-maximal stimulation at 1 ng/ml. The response of cells from normal and primary biliary cirrhotic liver to hHGF was similar. Cultures were immunostained with specific antibodies and then processed for [3H]thymidine autoradiography. Proliferating cells expressed BEC-specific markers (HEA125 and CK-19), but were negative for desmin and factor VIII-related antigen. Occasional vimentin-positive cells were observed, but these were nonproliferative. In conclusion, cells responding to hHGF were clearly BEC in origin. The observation that HGF is mitogenic for BEC as well as hepatocytes has important implications. First, greater yields of intrahepatic BEC are available for subsequent studies of the pathogenesis and etiology of diseases of the biliary epithelium. Secondly, some means of regulating the cellular response to HGF in vivo must operate, in that HGF levels rise early after partial hepatectomy and yet BEC proliferate 24 h later than hepatocytes.

Molecular analysis of the Pseudomonas aeruginosa genes, ruvA, ruvB and ruvC, involved in processing of homologous recombination intermediates.

In Escherichia coli, the products of the ruvA, ruvB and ruvC genes are all involved in the processing of recombination intermediates (Holliday structures) into recombinant molecules. We cloned a 9.4-kb DNA fragment from Pscudomonas aeruginosa PAO1 in a plasmid by functional complementation of the UV sensitivity of an E. coli strain with ruvABC deleted. In P. aeruginosa, the ruv region seemed to form a non-SOS regulated single operon consisting of orf26-ruvC-ruvA-ruvB, while in this region of E. coli, ruvA and ruvB form an SOS-regulated operon, orf26 and ruvC form a non-SOS operon, and these two operons are split by orf23. The deduced amino acid sequences of P. aeruginosa RuvA, RuvB and RuvC proteins were 55, 72 and 55% identical to those of the corresponding E. coli Ruv proteins. The individual ruv genes of P. aeruginosa complemented the corresponding single ruv mutations of E. coli, suggesting that the P. aeruginosa Ruv proteins can interact functionally with their E. coli Ruv partners in forming heterologous complexes. The sequence alignments of the Ruv proteins were extended by incorporation of data about the putative ruv genes obtained from data banks, and the RuvB sequences were conspicuously more conserved than the RuvA and RuvC sequences.

Amino acid sequence of alkaliphilic serine protease from silkworm, Bombyx mori, larval digestive juice.

Alkaliphilic protease, P-IIc, from silkworm, Bombyx mori, larval midgut digestive juice consists of 232 amino acids. It has a catalytic triad, Asp-His-Ser, invariably found in a serine protease. A shift of optimal pH value towards the alkaline side diminished at mu = 1.0. This suggests the existence of an electrostatic interaction that affects the proteolytic activity. The higher Arg content may be responsible for this phenomenon. Two cysteine residues probably exist unpaired in a novel position among serine proteases.

Isolation of several cDNAs encoding yeast peroxisomal enzymes.

Several candidate clones carrying partial cDNAs for yeast peroxisomal enzymes, such as catalase, carnitine acetyltransferase, isocitrate lyase, malate synthase and acyl-CoA oxidase, were efficiently isolated at a single plating from a phage lambda gt11 recombinant cDNA library prepared with poly(A)-rich RNA from an n-alkane-grown yeast, Candida tropicalis, with a mixture of antibodies against the respective purified enzymes. Among them, one candidate clone carrying partial cDNA for catalase was subcloned and subjected to nucleotide sequence analysis. We succeeded in determining that the amino acid sequence deduced from the nucleotide analysis included the sequences derived from the two peptide fragments obtained from the purified enzyme.

Inconsistent images of thyroid nodule scintigrams made with iodine and pertechnetate: case report.

In a patient with Hürthle cell adenoma, thyroid scans showed nodular trapping of 99mTc-pertechnetate, but nodular binding of 131I was not observed.

Presence of two transcribed malate synthase genes in an n-alkane-utilizing yeast, Candida tropicalis.

The presence of two genomic DNA regions encoding malate synthase (MS) was shown by Southern blot analysis of the genomic DNA from an n-alkane-assimilating yeast, Candida tropicalis, using a partial MS cDNA probe, in accordance with the fact that two types of partial MS cDNAs have previously been isolated. This was also confirmed by the restriction mapping of the two genes screened from the yeast lambda EMBL library. Nucleotide sequence analysis of the respective genomic DNAs, named MS-1 gene and MS-2 gene, revealed that both regions encoding MS had the same length of 1,653 base pairs, corresponding to 551 amino acids (molecular mass of MS-1, 62,448 Da; MS-2, 62,421 Da). Although 29 nucleotide pairs differed in the sequences of the coding regions, the number of amino acid replacements was only one: 159Asn (MS-1)----159Ser (MS-2). In the 5'-flanking regions, there were replacements of four nucleotide pairs, deletion of one pair, and insertion of four pairs. In spite of the fact that two genomic genes were present and transcribed, RNA blot analysis demonstrated that only one band (about 2 kb) was observable even when the carbon sources in the cultivation medium were changed. A comparison of the amino acid sequences was made with MSs of rape (Brassica napus L.), cucumber seed, pumpkin seed, Escherichia coli, and Hansenula polymorpha. A high homology was observed among these enzymes, the results indicating that the protein structure was relatively well conserved through the evolution of the molecule.(ABSTRACT TRUNCATED AT 250 WORDS)

Peroxisomal isocitrate lyase of the n-alkane-assimilating yeast Candida tropicalis: gene analysis and characterization.

A genomic DNA clone encoding isocitrate lyase, a key enzyme of the glyoxylate cycle and a peroxisomal enzyme of the n-alkane-assimilating yeast Candida tropicalis has been isolated with a cDNA probe from the yeast lambda EMBL library. Nucleotide sequence analysis of the genomic DNA clone disclosed that the region coding isocitrate lyase had a length of 1,650 base pairs, corresponding to 550 amino acids (61,602 Da). RNA blot analysis demonstrated that only one kind of mRNA (2 kb) supposed to be transcribed from this gene was present in the cells. A comparison of the amino acid sequences was made with the isocitrate lyase of castor bean, one of the glyoxysomal enzymes, and the enzyme of E. coli. The isocitrate lyases of C. tropicalis and castor bean had high homology, and the presence of some amino acid stretches conserved in all three enzymes suggests that these might be involved in the catalysis of the common reaction. There was an insertion common to the isocitrate lyases of C. tropicalis and castor bean, which is of interest concerning their evolution. In the C-terminal region, a characteristic sequence similar to that previously proposed as the import signal to peroxisomes was present.

Hepatocyte growth factor stimulates liver regeneration and elevates blood protein level in normal and partially hepatectomized rats.

The effects of recombinant human hepatocyte growth factor (HGF) on liver growth and function of normal and partially hepatectomized rats have been examined. HGF was continuously administered into the jugular vein because it was rapidly eliminated from the plasma (t1/2 alpha; approximately 4.5 min) and degraded. In normal rats, the labeling index of hepatocytes was increased about 6 times by the administration of HGF. HGF also decreased the prothrombin time and increased the hepaplastin and serum albumin content. In 70%-hepatectomized rats, HGF stimulated liver regeneration and increased the level of blood proteins such as hepaplastin in a dose-dependent manner. The stimulation of serum protein level seemed to result from not only the increase of hepatic cell number but also the direct effect of HGF on the protein production in hepatocytes, because HGF rapidly enhanced the protein synthesis prior to the increase of cell number and increased the mRNA content of albumin in the liver in vivo. In addition, a combination of heparin with HGF further accelerated the effects of HGF described above, possibly due to the decrease of HGF clearance. These findings suggest that HGF accelerates both the hepatic regeneration and function in vivo, and that rhHGF is clinically expected to be a potent therapeutic agent in hepatectomy and liver injury.

Protective effect of dipyridamole against lethality and lipid peroxidation in liver and spleen of the ddY mouse after whole-body irradiation.

The effects of dipyridamole on radiation damage in the mouse were investigated. Dipyridamole (i.p. 2 mg/mouse) administered 1 h before exposure, protected against gamma-irradiation. Pretreatment significantly decreased the death rate at 30 days from 89 to 33% (p<0.001) after 9 Gy whole-body irradiation. LD50 at 30 days was increased from 6.67 to 7.65 Gy in the dipyridamole pretreated group. The level of thiobarbituric acid reactive substances (TBARS) in the liver and spleen, a measure of free radical initiated liver peroxidation, increased 155, 193, 195, and 236% of control (without irradiation) in liver, and 132, 146, 168, and 276% of control (without irradiation) in spleen on days 2, 4, 7, and 10 after 9 Gy of whole-body irradiation respectively. The TBARS levels in both liver and spleen 2 days after irradiation were reduced to 73 +/- 7 and 60 +/- 19% respectively after dipyridamole treatment (2 mg/mouse, i.p. injection 1 h before exposure). In electron microscopic studies, mitochondria and endoplasmic reticulum in the irradiated mouse liver were swollen, but otherwise appeared normal after dipyridamole treatment. These results suggest that dipyridamole has a protective effect on animal survival 30 days after 60Co gamma-irradiation and inhibits lipid peroxidation - which is thought to play a part in the radiation injury in mouse liver and spleen.

Clinical studies using radiocolloid in chyluria.

Very little radioactive colloid, injected subcutaneously is excreted in urine by the normal kidney. In chyluria patients, however, radiocolloid injected subcutaneously (not venously) is passed through in larger quantities in the urine to the control patients, because of the lymphatic communication with urinary tract. As far as we investigated, measurement of the radiocolloid in urine is a valid clinical test for chyluria patients.

Effect of dimethyl sulfoxide pretreatment on activities of lipid peroxide formation, superoxide dismutase and glutathione peroxidase in the mouse liver after whole-body irradiation.

We investigated the effects of dimethyl sulfoxide (DMSO) on radiation damage in the mouse. DMSO (i.p. 0.11 g/mouse) administered 30 min before exposure protected the mice from the gamma-whole body irradiation: the 30 days lethality was significantly decreased from 44% to 16% (P < 0.05). The contents of thiobarbituric acid reactive substances(TBA-RS) in the mouse liver increased linearly between days 2 and 10 after 9 Gy gamma ray irradiation. The TBA-RS contents in the liver on days 2 to 10 after irradiation were reduced by DMSO pretreatment. The irradiation decreased superoxide dismutase (SOD) activity in the liver on day 10. Decrease in SOD activity was prevented by DMSO pretreatment. In the electron microscopic study, the mitochondria in the irradiated mouse liver were swollen, but we could observe no change after DMSO pretreatment. The results suggest that DMSO has radioprotective effects, probably due to inhibition of lipid peroxidation.

Regional lung perfusion and ventilation with radioisotopes in cervical cord-injured patients.

In general, cervical cord-injured patients present with restrictive pulmonary dysfunction resulting from paralysis of the intercostal muscles. Vital capacity frequently decreases below 50% of that in normal subjects, and their respiratory pattern frequently includes paradoxical movement in which the intercostal spaces sink and the abdomen distends at inspiration. Ventilation scintigraphy using Xe-133 and pulmonary perfusion scintigraphy using Tc-99m macroaggregated albumin (MAA) were performed on nine cervical cord-injured patients and four normal subjects to investigate regional lung functions in the cervical cord-injured patients. Pulmonary perfusion scintigraphy, in which measurement was made in the supine position, revealed no differences between the patients and the normal subjects. The inhomogeneous ventilation/perfusion distribution was presumed to have resulted from change in regional intrapleural pressure due to paradoxical movement of the thoracic cage. Washing and washout times were prolonged by paralysis of the intercostal muscles. These phenomena were particularly apparent in the upper and middle lung regions where compensating action by movement of the diaphragm is small.

The use of multimedia in patient care.

A personal computer based system was constructed to assess the use of various forms of information (multimedia) in patient record keeping. A patient's file with his records kept in a multimedia fashion was made by using the system. We describe the hardware and software construction of the system together with the results and the memory requirements of each type of media. Potential usage of the system in the future is discussed especially in connection with the Picture Archiving and Communications Systems (PACS).

[A case of mediastinal malignant lymphoma; 16 years follow up after surgery].

A 32-year-old female was admitted to our hospital on September 1976 because of left mediastinal mass shadows on chest roentgenogram. Needle biopsy studies provided no definitive diagnosis, and exploratory operation was carried out through left postero-lateral thoracotomy. Two large masses were seen in the mediastinum and five small tumors were seen on the diaphragm. All of these masses were removed. Histopathological examination of the tumors indicated non-Hodgkin's lymphoma, diffuse small cell type. Radiation therapy was carried out postoperatively, but chemotherapy could not continue because of side effect. Eight years after surgical therapy, recurrence was seen at left parietal pleura, ten years at peritoneum, twelve years at left parietal pleura, thirteen years at upper mediastinal lymph node, 16 years at post-peritoneal space. These tumors disappeared after radiation therapy. She is doing well seventeen years after the surgery.

[A case report of leiomyosarcoma of the inferior vena cava].

A case of leiomyosarcoma of the I.V.C. is described, with emphasis on the CT appearance. CT is useful for the diagnosis and follow up on this case.