[Video-assisted thoracic surgery for clinical stage I lung cancer].

We evaluated our results of video-assisted thoracic surgery (VATS) performed for lung cancer over 8 years. Between April 2000 and October 2008, a total of 409 (60.9%) underwent VATS for lung cancer. Operative procedures as a radical operation were partial resection in 58 patients, segmentectomy in 64 patients, and lobectomy in 229 patients. There was 1 patient with operative death including hospital death due to pulmonary thromboembolism. In a median follow-up period of 21 months, the 5-year cause specific survival rate was 93.7%. According to operative procedures, the 5-year survival rate was 100% in patients underwent partial resection and segmentectomy, and 91.1% in patients underwent lobectomy. According to pathological stages, the 5-year survival rate was 98.8% in 289 patients with stage IA, 69.1% in 34 patients with stage IB, and 68.2% in 14 patients with stage IIIA. In conclusion, VATS lobectomy and VATS intentional limited resection can be performed with low mortality and good prognosis for clinical stage IA lung cancer patients.

Midkine as a prognostic biomarker in oral squamous cell carcinoma.

The aim of this study was to evaluate serum midkine (S-MK) concentrations as a prognostic tumour marker in oral squamous cell carcinoma (OSCC). We measured S-MK concentrations in patients with OSCC and healthy volunteers. In addition, we performed real-time quantitative reverse transcription-PCR analysis and immunohistochemistry with fresh tumour samples. To determine whether S-MK concentrations have prognostic value, we performed survival analyses with clinical information by using the log-rank test. Serum midkine concentrations were significantly higher in patients with OSCC than in healthy controls (P<0.001). Serum midkine concentrations were also significantly increased in early-stage OSCC compared with those of healthy individuals (P<0.001). In addition, immunohistochemistry allowed identification of overexpressed MK protein in OSCC tissues. MK mRNA showed higher expression in OSCC samples compared with normal mucosal samples. Patients in high S-MK groups showed a significantly lower 5-year survival rate compared with patients in low S-MK groups (P<0.05). The increased S-MK concentrations in early-stage OSCC were strongly associated with poor survival. Serum midkine concentrations may thus be a useful marker not only for cancer screening but also for predicting prognosis of OSCC patients.

Catalytic activity and arrangement of subunit polypeptides in rat liver cytochrome c oxidase as studied by proteolysis.

Cytochrome c oxidase from rat liver was incubated with various proteinases of different specificities and the enzymic activity was measured after various incubation times. A loss of catalytic activity was found after digestion with proteinase K, aminopeptidase M and a mitochondrial proteinase from rat liver. In each case the decrease in enzymic activity was compared with the changes in intensities of the polypeptide pattern obtained after sodium dodecyl sulfate polyacrylamide gel electrophoresis. The susceptibilities of the subunit polypeptides of the soluble cytochrome c oxidase to proteinases were very different. Whereas subunit I was most susceptible, subunits V--VII were rather resistant to degradation. From the relative inaccessibility of subunits V--VII to proteinases it is likely that these polypeptides are buried in the interior of the enzyme complex.

Intercellular cell adhesion molecule-1 regulates lymphocyte movement into intestinal microlymphatics of rat Peyer's patches.

The objective of this study was to determine whether specific adhesion molecules modulate lymphocyte movement from Peyer's patches into intestinal microlymphatics. The fluorochrome acridine orange was injected via a micropipette into Peyer's patches to fill lymphatics. The flux of labeled lymphocytes into intestinal microlymphatics was monitored with intravital fluorescence microscopy. The lymphatic microvessels in the perifollicular area of Peyer's patches were filled with lymphocytes, most of which remained within the lymphatics. Some lymphocytes became detached and were drained into intestinal lymph. Administration of antibodies directed against ICAM-1 significantly increased lymphocyte flux into interfollicular lymphatics. The immunohistochemical study showed intense ICAM-1 expression on the lymphocytes densely packed in the lymphatics surrounding follicles in Peyer's patches. A large number of lymphocytes are normally sequestered in the lymphatic network of Peyer's patches. This sequestration of lymphocytes is largely mediated by ICAM-1-dependent cell-cell interactions.

Low-temperature heat capacities of confined liquid benzene, implying the behavior of ordinary bulk liquids.

Isobaric heat capacities C p of benzene confined in silica MCM-41 mesopores with average diameters equal to and smaller than 2.9 nm were measured by precise adiabatic calorimetry. The confined benzene samples revealed no thermal anomaly due to crystallization/fusion and vitrified at low temperatures. The C p curves displayed a hump and a considerably quick decrease on the low-temperature side of the hump as the pore diameter increased. The enthalpy-relaxation effects observed on intermittent heating showed that the anomaly of the C p hump and quick decrease is not assigned to a glass transition. The bend in the temperature dependence of density reported previously was interpreted as corresponding to the quick decrease in C p . We concluded that the anomalous C p and density behaviors originated from the ordering/excitation in the configurational state, close to the ground state, of confined molecular aggregate and proposed a scenario that explains the general C p curves of ordinary bulk supercooled liquids in equilibrium at low temperatures below the glass-transition temperatures.

Nucleotide sequence and expression of the organomercurial-resistance determinants from a Pseudomonas K-62 plasmid pMR26.

pMRA17 cloned from Pseudomonas K-62 plasmid pMR26 specified the resistance to both organic and inorganic mercurials. DNA sequence of this broad-spectrum resistant mer operon was determined. The 5504-bp sequence includes six open reading frames (ORFs), five of which were identified as merR, merT, merP, merA and merB in order by analysis of deletion mutants and by comparison with the DNA and amino acid (aa) sequences of previously sequenced mer operons. The merB encoding organomercurial lyase showed a less identity than the other mer genes with those from other broad-spectrum resistance operons. The remaining ORF named merE, located between merA and merB, had no significant homology with the published mer genes and seemed to be a new gene which may involve in phenylmercury resistance. Induction experiments and maxicell analyses of the mer-polypeptides revealed that pMRA17 mer operon expressed mercurial-inducible phenotype and the merB and merE as well as the merA were under the control of MerR which could activate not only by mercuric ion but also by organomercurials.

Diazepines. 5. Synthesis and biological action of 6-phenyl-4H-pyrrolo[1,2-a][1,4]benzodiazepines.

A series of 6-phenyl-4H-pyrrolo[1,2-a][1,4]benzodiazepines (2) has been prepared with 2-phthalimidomethylfurans (12) and 1-phthalimidoalkane-2,5-diones (15) or 2,5-dimethoxy-2-phthalimidomethyltetrahydrofurans (16) as the key intermediates and subsequently evaluated for CNS activity. The structure-activity data generated indicate that, in general, introduction of the methyl and/or ethyl group(s) in the pyrrole ring and a chlorine atom at the ortho position of the 6-phenyl group increases the activity and that substitution of the above chlorine atom for a fluorine atom decreases the activity. 8-Chloro-6-(2-chlorophenyl)-1,3-dimethyl-4H-pyrrolo[,2-a][1,4]benzodiazepine (2p), the most potent among the compounds synthesized, was equipotent in taming and sedative activities to diazepam. The acute LD50 of 2p in mice was larger than 3000 mg/kg po.

Effects of arginine derivatives on soluble guanylate cyclase from neuroblastoma N1E 115 cells.

The effects of L-arginine (Arg) derivatives on soluble guanylate cyclase from neuroblastoma N1E 115 cells were examined. The Arg derivatives were modified at the -NH2, -COOH, C alpha-proton or guanidino group of Arg. Among the synthesized derivatives, eight compounds, i.e. the 5-(dimethylamino)-1-naphthalenesulfonyl (DNS) ones, especially N-cyclohexyl-2-(N-DNSamino)-5-guanidino-2-methylvaleramide and 1-[2-(N-DNSamino)-2-(2-imino-1,2,3,4,5,6-hexahydropyrimidin- 4-yl)acetyl]- piperidine, were found to inhibit the activity of crude guanylate cyclase in the 105,000 g supernatant fraction of the cell homogenate. The enzyme, partially purified by a column of Chelex 100 Na+, was also inhibited by these eight compounds. The mode of the inhibition was competitive. The Ki values were in the range of 2-8 microM for the enzyme in the 105,000 g supernatant fraction and 3-16 microM for the partially purified enzyme, in the presence of Mg2+ as a metal cofactor. In contrast, a new derivative, methyl 2-amino-5-guanidinovalerate (M Arg ME), as well as the Arg methyl ester (Arg ME) and Arg; were found to enhance the activity of the partially purified guanylate cyclase; KA values of M Arg ME, Arg ME and Arg were approximately 9, 4 and 3 microM respectively. From these results, the free guanidino group including 2-imino-1,2,3,4,5,6-hexahydropyrimidin-4-yl or 2-imino-1,2,3,4,5,6-hexahydropyrimidin-5-yl and modification of the --NH2 residue with a hydrophobic group such as DNS seemed to be essential for inhibition of the guanylate cyclase; however, the guanidino and --NH2 residue of Arg should be free for activation by these Arg derivatives.

Tryptophan residues of saccharifying alpha-amylase from Bacillus subtilis. A kinetic discrimination of states of tryptophan residues using N-bromosuccinimide.

Four tryptophan residues of saccharifying alpha-amylase from B. subtilis out of eleven in total are reactive towards N-bromosuccinimide (NBS), suggesting that they are on the surface of the enzyme. This is consistent with the results of solvent perturbation difference spectrophotometry with ethylene glycol. One of four tryptophan residues was clearly distinguished from the other three in reactivity with NBS by the stopped-flow method. This most reactive tryptophan residue was not protected from modification by substrates of analogs, indicating that the tryptophan is not located in the substrate binding site. One of the other three tryptophan residues, probably the second most reactive one, is considered to be related in some way to the glycosyl transfer in the reaction of the enzyme with maltose as a substrate.

Lack of involvement of merT and merP in methylmercury transport in mercury resistant Pseudomonas K-62.

To clarify whether the merT and merP genes play a role in the transport of methylmercury, we constructed a deletion plasmid, pMRD141 which lacked the genes conferring the organomercurial lyase and the mercuric reductase from plasmid, pMRA17 containing the entire broad-spectrum mercury resistance determinants of the 26 kb-plasmid from Pseudomonas K-62. Plasmid, pMRD141 showed hypersensitivity to Hg2+ but still expressed a normal sensitivity to methylmercury. The mercury-induced hypersensitive cells carrying pMRD141 took up appreciably more 203Hg2+ than the induced resistant cells with pMRA17 and the sensitive cells with cloning vector, Bluescript II SK(+) but no difference in the uptake of CH3(203)Hg2+ among these three strains was found. These results suggested that the merT and merP are only involved in the Hg2+ transport but do not participate in the transport of methylmercury.

Volumetric MRI measurements of the tree shrew hippocampus.

Protocols suitable for repeated magnetic resonance imaging (MRI) studies of the tree shrew's brain were established. This included the development of (i) a technique for prolonged inhalation anesthesia by endotracheal intubation; (ii) a reproducible fixation of the animal's head in a stereotaxic frame and finally (iii) the set-up of the hardware (rf coil) and software (MRI sequences) of the MRI system. The endotracheal intubation as well as the repeated and prolonged anesthesia showed no complications. The in vivo measurements of the tree shrew's hippocampal formation revealed a high reproducibility. Right and left hippocampal volume was determined as 85.2 mm3 +/- 8% and 87.4 mm3 +/- 10%, respectively. The utility of MRI in delineating alterations in brain anatomy was demonstrated in three animals receiving cortisol via the drinking water (5 mg/animal/day). After a 4-week treatment, in two of the three tree shrews a reduction in hippocampal volume was observed. Thus, the MRI protocols used here allow for repeated and non-invasive measurements of changes in hippocampal anatomy within the same animal and to monitor the temporal dynamics of structural alterations within this brain structure.

Effect of nitric oxide on L-[3H]glutamate binding to rat brain synaptic membranes.

Nitric oxide (NO), which is spontaneously generated from sodium nitroprusside, was shown to inhibit L-[3H]glutamate binding to rat brain synaptic membranes in a concentration-dependent manner. The L-glutamate binding inhibited by NO, was largely recoverable by the addition of hemoglobin, a scavenger of NO, to the assay medium. These results suggest that NO may play an important role in the modulation of excitatory neurotransmission through direct interaction of L-glutamate binding to its physiological synaptic membrane receptors.

Effects of glucocorticoids on apoptosis of infiltrated eosinophils and neutrophils in rats.

The effects of glucocorticoids on the survival of rat eosinophils and neutrophils infiltrated into the peritoneal cavity were examined. Glucocorticoids including dexamethasone, prednisolone and hydrocortisone inhibited the survival of rat peritoneal eosinophils at 10(-6) M, whereas they prolonged survival of rat peritoneal neutrophils at 10(-8) M. Sex steroids including estradiol and progesterone did not affect cell survival. Dexamethasone decreased the viability of eosinophils after 3 days of incubation and maintained the viability of neutrophils until 4 days after incubation concentration dependently. The EC50 of dexamethasone for inhibition of the survival of eosinophils was 1.5 x 10(-8) M, and that for the spontaneous death of neutrophils was 6.4 x 10(-10) M, suggesting that glucocorticoids at concentrations that inhibit eosinophil survival prolong neutrophil survival. Analysis of DNA fragmentation of cultured eosinophils and neutrophils revealed that glucocorticoids enhance eosinophil apoptosis but inhibit neutrophil apoptosis. The effects of dexamethasone on viability and DNA fragmentation were counteracted by the glucocorticoid receptor antagonist, mifepristone, concentration dependently. These findings indicate that glucocorticoids induce contradictory effects via the glucocorticoid receptor on rat eosinophils and neutrophils extravasated to an inflammatory locus such as the peritoneal cavity by modulating apoptosis.

Manganese-metalloporphyrin (ATN-10) as a tumor-localizing agent: magnetic resonance imaging and inductively coupled plasma atomic emission spectroscopy study with experimental brain tumors.

OBJECTIVE: We examined whether selective tumor accumulation of a novel manganese-metalloporphyrin (ATN-10) occurs in Fisher rats bearing intracerebral 9L gliomas. METHODS: After intravenous administration of ATN-10, magnetic resonance imaging of brains with tumors or nontumoral vasogenic brain edema was performed. Tissue manganese concentrations were measured by inductively coupled plasma atomic emission spectroscopy until 48 hours after administration of ATN-10, to evaluate its uptake in tumor, normal brain, and peritumoral brain tissue. RESULTS: In magnetic resonance imaging scans, early enhancement was observed in both tumor tissue and regions of nontumoral vasogenic brain edema at 5 minutes after ATN-10 administration. However, delayed enhancement was noted only in tumor tissue, at 24 hours after intravenous injection of ATN-10. Comparison of rat brain specimens and 24-hour magnetic resonance imaging scans revealed that only the viable portions of tumors were enhanced with ATN-10; necrotic regions and areas of peritumoral brain tissue and nontumoral vasogenic edema were not. Significantly greater uptake of ATN-10 was found in tumor samples, compared with normal and peritumoral brain tissue, at 24 hours. A high tumor/normal brain tissue ratio (10.4) was achieved at 24 hours. CONCLUSION: ATN-10, a manganese-metalloporphyrin, is a potentially useful tumor-localizing agent that accumulates and is preferentially retained in viable tumor tissue.

Cavernous hemangioma over the anterior fontanelle.

This is a report of an extremely rare case of cavernous hemangioma over the anterior fontanelle in a 5-year-old boy. Cavernous hemangioma is most common in or beneath the skin of the face, neck, and extremities and sometimes occurs in the scalp. Although cavernous hemangioma is often quoted in the literature as a differential diagnosis of a mass located at the anterior fontanelle, we could not find an illustrative case in published articles. We discuss the characteristics and differential diagnosis of cavernous hemangioma over the anterior fontanelle.

Effect of kallikrein-kinin system on ion transport across rat small intestine.

The flow of glucose and valine from the mucosal to the serosal side of rat intestine was stimulated significantly by kallikrein-kinin system. To elucidate the mechanism of this function the effects of kallidin on the transmural potential difference (PDt) between the serodal and the mucosal sides and the transepithelial resistance (Rt) of rat jejunum segment were investigated by electrophysiological procedures. The glucose-evoked PDt was not significantly altered by addition of kallidin to the mucosal fluid. Meanwhile, PDt which evoked by glucose was significantly shifted under the presence of 10-1-10(2) ng kallidin in the serosal fluid. The valine-evoked PDt was also changed by addition of 10 ng kallidin into the serosal fluid. However, the addition of glucose to the mucosal and kallidin to the serosal fluid did not affect on the Rt of the jejunum segment. These results suggest that kinin stimulated Na+ transport at the baso-lateral membrane of the intestinal absorptive cells, and the transports of other substances were enhanced consequently.

Purification and partial characterization of cat colon and submandibular gland kallikreins.

Kallikreins from cat colon and submandibular gland have been purified by acetone fractionation of tissue extracts, DEAE-Sephacel ion-exchange chromatography, rho-aminobenzamidine Sepharose 4B affinity chromatography and gel filtration on Sephadex G-75. They were of similar M.W., approximately 40,000, and each comprised five forms by isoelectric-focusing (pI 4.1-4.8). Both enzymes were potent kininogenases and exhibited similar specificities with synthetic ester and amide substrates. They were susceptible to a range of protease inhibitors. Surprisingly, neither was sensitive to aprotinin yet both were partially inhibited by soya-bean trypsin inhibitor. They were indistinguishable in our immunological tests. An acidic esterase (pI 2.2-3.5) of M.W. 120,000 was isolated from cat stomach by the same procedure. While it exhibited weak immunologic similarity to cat submandibular gland kallikrein, it had negligible kininogenase activity and different substrate and inhibitor specificities to the two kallikreins. It is concluded that similar tissue kallikreins are present in the colon and submandibular gland of the cat but are distinct from this cat stomach esterase.

Purification and partial characterization of cat pancreatic and urinary kallikreins--comparison with other cat tissue kallikreins and related proteases.

Kallikreins have been purified from cat pancreas and urine by methods similar to those described previously for cat colon and submandibular gland kallikreins. The pancreatic kallikrein (M.W. 41,200, pI 4.75) was similar to the urinary kallikrein (M.W. 34,300 pI 4.35-4.70) in pH optimum, substrate specificity and inhibition profile. Both enzymes were potent kininogenases and immunologically similar. These enzymes closely resembled the kallikreins from cat colon and submandibular glands. A trypsin (M.W. 18,800) was isolated from cat pancreas and shown to be distinct from the group of kallikreins in all parameters tested. We attempted purification of cat renal kallikrein, but were unable to isolate any such enzyme. The major acidic esterase of cat kidney cortex (M.W. 59,000, pI 4.91) was purified and was distinct from both the cat tissue kallikreins and trypsin. The origin of cat urinary kallikrein remains unclear, but in the light of our findings, it may result from renal filtration of blood-borne tissue kallikreins rather than from intrarenal synthesis.

Cytotoxic effect of sodium nitroprusside on PC12 cells.

To investigate the biochemical mechanism responsible for NO-induced neurotoxicity, the effect of sodium nitroprusside(SNP), a NO-generating agent, on PC12 cells was studied. The cell density was dose-dependently inhibited by SNP. Neuronally differentiated PC12 cells showed a higher resistance to SNP than the undifferentiated cells. The inhibitory effect was enhanced by 8-Br-cGMP, and reduced by methylene blue. However, 8-Br-cGMP alone had no significant cytotoxicity. SNP also inhibited [3H]-thymidine incorporation into the cells in a dose-dependent manner. The dose response curves for reducing cell density and for inhibiting thymidine incorporation, were found to be virtually superimposable. These results suggested that cytotoxicity elicited by NO seemed to be due to inhibition of DNA synthesis in PC12 cells.

Tumor specific contrast enhancement study of Mn-metalloporphyrin (ATN-10)--comparison of rat brain tumor model, cytotoxic and vasogenic edema models.

ATN-10, Mn-metalloporphyrin, has been developed as a tumor selective contrast agent for magnetic resonance (MR) imaging. To investigate the tumor specificity of ATN-10, we produced three experimental in vivo models; rat bran tumor (9L glioma) model, vasogenic (cold injury) and cytotoxic brain edema (24-hour MCA occlusion) models. The time course of contrast enhancement was compared after intravenous injection of ATN-10 or Gd-DTPA, measuring the signal intensity of the region of interest. After ATN-10 administration, the 9L glioma model showed early (5 min) and delayed (24 hr-) peak enhancement whereas the cold injury model showed only early enhancement and the 24-hour MCA occlusion model did not show significant enhancement. After Gd-DTPA administration, all three models showed similar pattern of only early enhancement. As a contrast agent for MR imaging, ATN-10 showed different behavior than Gd-DTPA in demonstrating the blood-brain barrier disruption and moreover ATN-10 showed selective enhancement in experimental brain tumors.