[Quality of life of long-term survivors of surgically treated lung cancer].

Quality of life (QOL) of long-term survivors (more than 3 years after surgery) of primary non-small cell lung cancer was studied. QOL was analyzed using European Organization for Research and Treatment of Cancer Quality of Life Questionnaire, 30-Item version 3.0 (QLQ-C30) and Hospital Anxiety and Depression Scale (HADS). Sixty of 91 patients (66%) participated in this study 87 +/- 5 (38-172) months postoperatively. In QLQ-C30, calculated scores of physical (84.0 +/- 2.4), role (81.3 +/- 3.6), cognitive (79.7 +/- 2.6), emotional (86.8 +/- 1.9), and social (91.0 +/- 1.9) functioning, and global QOL (72.6 +/- 2.9) were obtained. Calculated HADS A (anxiety) was 3.3 +/- 0.3 and HADS D (depression) was 4.0 +/- 0.4. Postoperative follow-up duration was correlated with financial impact only. QOL of long-term survivors was influenced by gender histology, marital status, employment status, and academic carrier.

[Chondrosarcoma of rib origin; report of a case].

Chondrosarcoma of rib origin is rare. A 50-year-old man without symptom was pointed out an abnormal shadow on chest X-ray. Computed tomography (CT) showed a low density mass arising from the right chest wall, and a CT-guided needle aspiration biopsy disclosed the tumor consisted of cartilage matrix with a partial necrosis. We suspected the tumor to be a chondrosarcoma of rib origin and performed a wide resection with the right 3rd and 4th ribs. The defect of the chest wall was repaired with double prolene mesh. Histological examination revealed grade 2 chondrosarcoma. Postoperative course has been uneventful for 25 months.

[Changing pattern of the respiratory function associated with resected lung lobe].

Respiratory function before and 2 months after lung lobectomy was analyzed associated with resected lobe. Post- or preoperative ratios of FEV1.0 or VC were compared among (1) predicted value by the number of subsegments using bronchofiberscopy, (2) predicted value by the lobar volume ratio using computed tomography (CT), and (3) actually measured value. Using subsegments method, post- or preoperative predicted VC ratios were 85 +/- 1% after right upper lobectomy (RU), 69 +/- 1% after right lower lobetomy (RL), 74 +/- 1% after left upper lobectomy (LU), and 75 +/- 1% after left lower lobectomy (LL). Using CT method, post- or preoperative predicted VC ratios were 80 +/- 2% after RU, 76 +/- 4% after RL, 74 +/- 2% after LU, and 79 +/- 3% after LL. Actually measured post- or preoperative FEV1.0 ratios were 82 +/- 3% after RU, 89 +/- 8% after RL, 73 +/- 3% after LU, and 86 +/- 5% after LL, and the VC ratios were 88 +/- 5% after RU, 79 +/- 3% after RL, 77 +/- 4% after LU, and 94 +/- 3% after LL. In the FEV1.0 analysis using both subsegments method and CT method, the predicted value was correlated with upper lobectomy but was overestimated in case of lower lobectomy. This phenomenon might be caused by the postoperative bronchial branching deformity after upper lobectomy. In the VC analysis using subsegments method, the predicted value was correlated with upper lobectomy but was overestimated in case of lower lobectomy. Meanwhile, in the VC analysis using CT method, the predicted value was correlated with RL or LU but was overestimated in case of RU or LL. This may due to the fact that RL and LU had large lobar volumes. In conclusion, postoperative predicted and actually measured values were different associated with resected lobe. In the FEV1.0 and VC analysis using subsegments method, the predicted value was strongly correlated with upper lobectomy but was overestimated (10%) in case of lower lobectomy.

[Experience with invasive thymoma presenting pleural dissemination].

A 61-year-old man was admitted to Showa University Hospital because of a myasthenia gravis. Chest computed tomography revealed a mediastinal invasive tumor. During surgery, invasion to the pericardium and dissemination on the left visceral pleura and the left diaphragm were observed. Extended thymo-thymectomy and partial resection of the pericardium, left lung, and diaphragm were performed. Incomplete resection was achieved because of the dissemination on the diaphragm. Chemotherapy using ADOC and radiotherapy for mediastinum and left diaphragm were done. Four years after surgery, neither recurrence of the tumor nor myasthenia gravis was observed.

Comparison of the expression of cell surface poly-N-acetyllactosamine-type oligosaccharides in PC12 cells with those in its variant PC12D.

To explore the biological role of carbohydrate chains in the process of nerve cell differentiation, we carried out a characterization of the carbohydrate structure of glycoproteins by comparing conventional PC12 cells with variant cells (PC12D). In vitro metabolic labeling of cells with either [(3)H] glucosamine or [(3)H] threonine, together with tomato lectin staining, revealed that nerve growth factor (NGF) stimulation caused a decrease in the poly-N-acetyllactosamine synthesis of high-molecular-weight glycopeptides from PC12 cells. By comparison, the amount of glycopeptides with poly-N-acetyllactosamine from PC12D cells was already significantly low and it was not changed by NGF stimulation. By assaying the glycosyltransferases that participate in poly-N-acetyllactosamine synthesis, the decrease in the amount of the poly-N-acetyllactosamine in PC12D cells as well as NGF-stimulated PC12 cells could be accounted for by a reduction in the activity of poly-N-acetyllactosamine extension enzyme (GnT-i), because the amount of poly-N-acetyllactosamine in both cells precisely correlated with changes in GnT-i activity, whereas the activities of N-acetylglucosaminyltransferase V (GnT-V) and beta 1-4 galactosyltransferase remained unchanged. These results demonstrate that the decrease in poly-N-acetyllactosamine synthesis in PC12 cells occurred prior to neurite formation, whereas PC12D cells were insensitive to this effect. Next, we showed that GnT-i but not GnT-V catalyzed a rate-limiting reaction in the expression of poly-N-acetyllactosamine chains, especially in pheochromocytoma.

Keratin mRNA for detecting micrometastasis in cervical lymph nodes of oral cancer.

We studied three keratin (K) gene candidates, K13, K19, and K20 mRNAs, for detecting micrometastases in cervical lymph nodes (LNs) by reverse transcriptase-polymerase chain reaction (RT-PCR). Of 166 histologically metastasis-negative nodes, 24 micrometastatic LNs (14. 4%) were detected based on K13 gene expression. Keratin 19 mRNA is an inadequate marker for the genetic diagnosis due to not only illegitimate gene expression from lymphatic tissue but also gene expression from the ectopic salivary gland. Keratin 20 mRNA showed low sensitivity. It is suggested that K13 mRNA may be a promising tumor marker among these keratin genes for detecting the micrometastases in cervical LNs of oral cancer.

Gene expression of GLUT isoforms and VHL in oral squamous cell carcinoma.

In human oral squamous cell carcinoma (OSCC) cell lines, we detected atypical mRNA expression of GLUT2 and/or GLUT4 in addition to enhanced expression of GLUT1 mRNA using RT-PCR. In semi-quantitative reverse transcription-polymerase chain reaction analysis of mRNA expression in OSCC cell lines, we found an inverse relationship between mRNA expression of von Hippel-Lindau (VHL) and that of GLUT1, with no apparent influence on the expression of other GLUTs. These findings suggest that the reduction of VHL may play a critical role in glucose uptake of OSCC cell lines, with enhancement of GLUT1 expression.

Indication for epidural morphine for the relief of intractable pain in advanced oral cancer: report of four cases.

It can be difficult to manage the pain of advanced oral cancer. We present four patients in whom epidural morphine was used for intractable pain at primary or metastatic sites. For pain supplied by the trigeminal or cervical nerve a small dose of morphine was given through an epidural catheter inserted into the epidural space through C7-Th1. A favourable clinical response was achieved in three. In particular, in one patient who was given continuous morphine using a computerized ambulatory drug delivery system, we achieved excellent efficacy and stable control of pain. We think that the effect of the epidural morphine was decreased in the patient who did not respond because he had previously been treated with high oral doses. The present study confirmed that morphine given epidurally in small doses has a strong and prolonged analgesic action with less toxicity than when given orally.

Cytokeratin expression in squamous cell carcinoma of the lung and oral cavity: an immunohistochemical study with possible clinical relevance.

OBJECTIVE: Serologic identification of the fragment of cytokeratin 19 known as CYFRA 21-1 has been used for early detection of squamous cell carcinoma of the lung. The sensitivity of the CYFRA 21-1 assay in detecting oral cancers is lower than that in detecting lung cancers. To clarify the reason for this, we compared the cytokeratin expression in these cancers, with special reference to cytokeratin 19. STUDY DESIGN: Oral squamous cell carcinomas and lung squamous cell carcinomas were immunostained with cytokeratin 19, cytokeratin 10, and cytokeratin 13 antibodies. Staining intensity was scored on a graduated scale from 0 to 4. RESULTS: With respect to cytokeratin 19, the stainings of all lung cancers were scored as 4, which indicates a greater expression of cytokeratin 19 than is seen in oral cancers (p < 0.01). With an average cytokeratin 19 staining score of 1.67, oral cancers ranked lowest among the antibodies. Squamous cell carcinomas of the maxillary sinus arising from pseudostratified ciliated epithelium were highly expressive of cytokeratin 19. A marker for keratinizing cells (cytokeratin 10) and a marker for squamous cells (cytokeratin 13) were expressed more frequently and intensely in oral cancers (p < 0.01) than in lung cancers (p = 0.019). CONCLUSIONS: From the viewpoint of immunohistochemistry, cytokeratin 19 was found to be a tumor marker with low specificity and sensitivity in oral cancers. The staining results suggested that poor expression of cytokeratin 19 by oral squamous cell carcinoma may result in a low serum value of CYFRA 21-1 in patients with this condition.

Calcein is excreted from the intestinal mucosal cell membrane by the active transport system.

In this study, we report the efflux mechanism of calcein, an organic anion, mediated by a multidrug resistance-associated protein (MRP)-like protein in the intestinal mucosal membrane. The transport of calcein from the mucosal to serosal side was decreased dose-dependently and was significantly lower than that of the opposite direction. In addition, its transport was increased in the presence of metabolic inhibitors and probenecid. Furthermore, the efflux of calcein from the intestinal cell membrane, which was preloaded with calcein acetoxymethyl ester, was predominantly observed in the mucosal side rather than in the serosal side. Its efflux to the mucosal side was inhibited by the metabolic inhibitors and probenecid, not by verapamil which is a P-glycoprotein substrate. These results indicated that the transport of calcein and possibly other organic anions across the intestinal membrane may be regulated by the MRP-like protein, but not P-glycoprotein.

High glucose-induced cytosolic phospholipase A2 activation responsible for eicosanoid production in rat mesangial cells.

The stimulation of prostaglandin E2 (PGE2) production in mesangial cells exposed to a high glucose level was studied from the viewpoint of its implication in the glomerular hyperfiltration in diabetic nephropathy. The basal PGE2 synthesis apparently increased in the cells on incubation with a high glucose level (20 mM) for 3-6 h. Under these conditions, secretory phospholipase A2 activity was not detected in the incubation medium, but cytosolic phospholipase A2 (cPLA2) activity in the cells increased time-dependently up to 6 h, compared with that with a normal glucose level (5 mM). However, no difference in the cPLA2 protein content between the two glucose levels was observed on immunoblot analysis, suggesting that the increased cPLA2 activity under high glucose conditions is not due to stimulation of de novo synthesis. Stimulation with a calcium ionophore markedly enhanced arachidonic acid liberation and PGE2 production by cells exposed to the high glucose level. Furthermore, mitogen-activated protein kinase (MAPK) activity increased time-dependently under high glucose conditions, the rate of increase being consistent with those in cPLA2 activity and PGE2 production under the same conditions. These data suggest that glucose-induced cPLA2 activation through MAPK activation is responsible for the enhancement of PGE2 production in mesangial cells.

Endotoxin-induced enhancement of glucose influx into murine peritoneal macrophages via GLUT1.

Hypoglycemia is among the most injurious metabolic disorders caused by endotoxemia. In experimental endotoxemia with lipopolysaccharide (LPS) in animals, a marked glucose consumption is observed in macrophage-rich organs. However, the direct effect of LPS on the uptake of glucose by macrophages has not been fully understood, and the present study was undertaken to shed light on this point. The consumption and uptake of glucose, as measured with 2-deoxy-D-[3H]glucose, by murine peritoneal exudate macrophages in culture were accelerated two- to threefold by stimulation with 3 ng of LPS per ml. The rate of glucose uptake reached a plateau after 20 min of stimulation and remained at the maximum as long as LPS was present. Northern (RNA) blot analysis with cDNA probes for five known isoforms of glucose transporter (GLUT) revealed that the expression of GLUT by macrophages was restricted to the GLUT1 isoform during LPS stimulation and the amount of GLUT1 mRNA was increased by the stimulation. These results suggest that macrophage responses to LPS are supported by a rapid and sustained glucose influx via GLUT1 and that this is a participating factor in the development of systemic hypoglycemia when endotoxemia is prolonged.

Complete primary structure and phosphorylation site of the 65-kDa macrophage protein phosphorylated by stimulation with bacterial lipopolysaccharide.

There is ample evidence that intracellular protein phosphorylation is a mandatory event in the process of macrophage activation by LPS, yet how this event is initiated and what roles the phosphorylated proteins are assigned to are poorly understood. We previously isolated a 65-kDa cytosolic protein (pp65) that was phosphorylated specifically in LPS-stimulated murine macrophages. In the present study, the complete primary structure of pp65 was determined on the basis of the cDNA containing an open reading frame of 1881 bases. The sequence of pp65 revealed that it is a murine homologue of human L-plastin, recently identified as a novel transformation-induced polypeptide of neoplastic human cells, and that it contains a unique series of Ca2+, calmodulin, and actin binding domains. A single phosphorylated peptide was isolated from the tryptic digest of pp65 by reverse-phase HPLC. From the amino acid sequence of the dodecapeptide Gly-Ser-Val-Ser-Asp-Glu-Glu-Met-Met-Glu-Leu-Arg, the phosphorylation site of pp65 was located at the N-terminal region adjacent to the first Ca2+ binding domain. This sequence contains a repeat of the casein kinase II motif Ser-Xxx-Xxx-Glu/Asp and, together with the preceeding Arg residue, constitutes the consensus sequence Arg-Xxx-Ser for cAMP-dependent protein kinase (PKA) and protein kinase C (PKC), but not mitogen-activated protein kinase (MAPK)-specific motif is found. These results, taken together with previous observations on the process of macrophage activation by LPS, demonstrate that pp65 is phosphorylated by an LPS-induced protein kinase other than MAPK and exerts its function on the cytoskeleton in a Ca2+/calmodulin-dependent manner.

Staphylococcus aureus Wood 46 strain activates human B cells without affecting DNA synthesis or tyrosine phosphorylation.

Induction of Ig secretion in human tonsilar B cell by protein A-deficient Staphylococcus aureus WOOD 46 strain (SAW) was examined. SAW induced as much Ig secretion as protein A-rich S. aureus Cowan I strain (SAC) when IL-2 was present in the culture. Activated and resting B cells are separated by Percoll gradient to determine whether SAW stimulates either resting or activated B cells. In resting B cells, SAW plus IL-2 induced IgM secretion significantly, but neither IL-2 nor SAW alone induced IgM secretion. In activated B cells, however, IL-2 induced IgM secretion by itself and SAW plus IL-2 did not induce additional IgM secretion. These results suggest that SAW activates small resting B cells rather than preactivated B cells. Subsequently, mechanisms of B cell activation by SAW and SAC were compared. SAW did not induce [3H]-TdR incorporation through day 1 to 5, and the number of viable cells was not increased by SAW stimulation. Moreover, SAW did not induce significant tyrosine phosphorylation at any concentration tested, when tyrosine phosphorylation of B cells was examined. However, SAC induced both [3H]-TdR incorporation and tyrosine phosphorylation of B cell efficiently. In further experiments, induction of IL-2R and IgM mRNA expressions were examined. SAW by itself induced IL-2R and IgM mRNA expressions without affecting expression of membrane-type IgM mRNA. These results show that SAW activates human B cells in quite a different manner from SAC by up-regulating the expression of secretory type IgM mRNA without affecting cell proliferation nor tyrosine phosphorylation, proposing a distinct B cell activation model from SAC.

Synthesis of porphinatoirons having an alkyl amphiphilic chain and their O2 binding properties in lipid bilayers.

Synthesis and characterization of two amphiphilic tetraphenylporphinatoiron complexes having a glycerophosphocholine or an alkyl phosphoserine group are described. These porphinatoiron(II) complexes with 1-dodecyl-2-methyl-imidazole (L2MIm) were efficiently embedded in the bilayer of a phospholipid vesicle due to their high compatibility with lipids, similar to the heme substituted with four alkyl amphiphilic chains (lipid-heme). Oxygen transporting ability of the phospholipid vesicles embedded with hemes were similar to that of hemoglobin (Hb) in red blood cells.