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Background: It was previously reported that a functional human (h) Rad9 protein accumulated in the nuclei of non-small cell lung carcinoma (NSCLC) cells. Those experiments, however, did not examine whether the hRad9 gene was mutated in those cells. The sequence of the HRAD9 gene in NSCLC cells was investigated., Methods: The sequence of the HRAD9 was examined in tumor and peripheral normal lung tissues obtained from 50 lung adenocarcinoma patients during surgery. The expression of its mRNA using reverse transcription polymerase chain reaction (RT-PCR) was also examined., Results: No sequence alterations were detected in the HRAD9 gene, which was found to be normally transcribed in surgically resected lung carcinoma cells. However, in eight (16.0%) cases a single nucleotide polymorphism (SNP) was observed at the second position of codon 239 (His/Arg heterozygous variant) of the gene. This frequency was significantly higher than that found in the normal population., Conclusions: Whereas the capacity to produce a functional hRad9 protein was intact in lung adenocarcinoma cells, a nonsynonymous SNP of HRAD9 was detected that might be associated with the development of lung adenocarcinoma.

Selective oxidation of single-wall carbon nanotubes (SWCNTs) by H2O2 was conducted at varying heating times and monitored by UV-vis-NIR spectroscopy. A major increase in the relative absorption intensity indicated a higher than 80% concentration of metallic SWCNTs in the final product. Here, it is suggested that semiconducting SWCNTs are more reactive than metallic SWCNTs because of hole-doping by H2O2, resulting in faster oxidation.

E1B55K adenovirus codes the protein that inactivates the p53 protein of host cells and facilitates its own cell proliferation. We prepared E1B55K-deleted adenovirus and investigated its cytopathic effects in primary lung cancer cells to evaluate the possibility of its application in gene therapy. A p53 wild-type small cell lung cancer cell line (SBC3) and three p53 mutant cell lines (SBC5, small cell lung cancer; PC3, adenocarcinoma; and EBC1, squamous cell carcinoma) were infected with AxE1AdB, in which the E1B55K gene was deleted, to assess the cytotoxicity and induction of apoptosis. Moreover, the mRNA expression in virus-infected cells was studied with cDNA array analysis to investigate the response of cancer cells to the virus infection. As a result, AxE1AdB had no detectable cytopathic effect on SBC3 cells, and SBC3 cells continued to replicate. In contrast, AxE1AdB killed SBC5, PC3, and EBC1 cells with great efficiency. In SBC3, apoptosis of the host cells was observed in the early infection stage; in the p53-mutant cell lines, apoptosis of the cells was restricted. An analysis of gene expression in the host cells indicated a significant increase of Bax mRNA in SBC3 and its deficiency in SBC5. Overall, our results suggest that AxE1AdB replicated selectively in the p53-mutant lung cancer cells and showed strong cytotoxicity against them. In the p53-intact cells, p53 induced apoptosis through Bax expression and prevented virus replication and escalation of the infection.

Background: DNA damage sensor proteins have received much attention as upstream components of the DNA damage checkpoint signaling pathway that are required for cell cycle control and the induction of apoptosis. Deficiencies in these proteins are directly linked to the accumulation of gene mutations, which can induce cellular transformation and result in malignant disease., Methods: Using 48 sets of tumor tissue specimens and peripheral normal lung tissue specimens from 48 patients with nonsmall cell lung carcinoma (NSCLC) who underwent surgery, the authors investigated the expression of hRad9 protein, a member of the human DNA damage sensor family, using immunohistochemical and Western blot analyses., Results: Immunohistochemical analysis detected the accumulation of hRad9 in the nuclei of tumor cells in 16 tumor tissue specimens, (33% of tumor tissue specimens examined). Western blot analysis also revealed elevated levels of phosphorylated hRad9 protein in NSCLC cells that was accompanied by the detection of phosphorylated Chk1, a protein kinase that regulates the downstream signaling of the DNA damage checkpoint pathway. Furthermore, strong expression of hRad9 was correlated with an increase in Ki-67 expression index in the tumor cells that were examined., Conclusions: The findings made in the current study suggest that Rad9 expression may play an important role in cell cycle control in NSCLC cells and may influence NSCLC cell phenotype.

The linkage of sister chromatids after DNA replication ensures the faithful inheritance of chromosomes by daughter cells. In budding yeast, the establishment of sister chromatid cohesion requires Ctf8, Dcc1, and Ctf18, a homologue of the p140 subunit of the replication factor C (RFC). In this report we demonstrate that in 293T cells, Flag-tagged Ctf18 forms a seven-subunit cohesion-RFC complex comprised of Ctf18, Dcc1, Ctf8, RFCp40, RFCp38, RFCp37, and RFCp36 (Ctf18-RFC). We demonstrate that a stoichiometric heteroheptameric Ctf18-RFC complex can be assembled by coexpressing the seven proteins in baculovirus-infected insect cells. In addition, the two other stable subcomplexes were formed, which include a pentameric complex comprised of Ctf18, RFCp40, RFCp38, RFCp37, and RFCp36 and a dimeric Dcc1-Ctf8. Both the five- and seven-subunit Ctf18-RFC complexes bind to single-stranded and primed DNAs and possess weak ATPase activity that is stimulated by the addition of primed DNA and proliferating cell nuclear antigen (PCNA). These complexes catalyzed the ATP-dependent loading of PCNA onto primed and gapped DNA but not onto double-stranded nicked or single-stranded circular DNAs. Consistent with these observations, both Ctf18-RFC complexes substituted for the replicative RFC in the PCNA-dependent DNA polymerase delta-catalyzed DNA replication reaction. These results support a model in which sister chromatid cohesion is linked to DNA replication.

Tumor necrosis factor-alpha (TNF-alpha) induces both rapid onset of apoptosis and monocytic differentiation in HL-60 human myeloid leukemia cells. In this study, we examined the effect of activation of protein kinase C (PKC) in c-Myc protein expression in association with the induction of apoptosis and differentiation in TNF-alpha-treated HL-60 cells. Pretreatment with phorbol 12-myristate 13-acetate (PMA), an activator of PKC, prevented TNF-alpha-induced rapid onset of apoptosis, which occurs at 3 h culture with TNF-alpha, concomitantly with the up-regulation of c-Myc protein expression. In addition, PMA enhanced TNF-a-induced differentiation at 24h treatment. This was documented by the expression of integrin Mac-1 molecule (CD11b) on the cell surface and the cellular adhesion to the plastic bottom of the flask, indicating the differentiation along with the monocyte/macrophage lineage. These results indicate that activation of PKC not only counteracts apoptosis but also enhances differentiation in TNF-alpha-treated HL-60 cells. Up-regulation of c-Myc protein evoked by pretreatment with PMA for a short time could disturb the signaling pathway of the ceramide and sphingosine, which are known to function as the endogenous modulators mediating the apoptotic signal of TNF-alpha. Our results strongly suggest the role of c-Myc protein as a mediator of cytoprotective effect of PKC pathway, and PKC pathway opposes apoptosis and consequently undergo differentiation via rapid up-regulated c-Myc protein expression during TNF-alpha signaling of HL-60 cells. Our findings provide a new insight for a role of PKC and c-Myc protein with special reference to the regulatory mechanisms in the decision of cellular fate, differentiation or apoptosis.

The human DNA damage sensors, Rad17-replication factor C (Rad17-RFC) and the Rad9-Rad1-Hus1 (9-1-1) checkpoint complex, are thought to be involved in the early steps of the DNA damage checkpoint response. Rad17-RFC and the 9-1-1 complex have been shown to be structurally similar to the replication factors, RFC clamp loader and proliferating cell nuclear antigen polymerase clamp, respectively. Here, we demonstrate functional similarities between the replication and checkpoint clamp loader/DNA clamp pairs. When all eight subunits of the two checkpoint complexes are coexpressed in insect cells, a stable Rad17-RFC/9-1-1 checkpoint supercomplex forms in vivo and is readily purified. The two individually purified checkpoint complexes also form a supercomplex in vitro, which depends on ATP and is mediated by interactions between Rad17 and Rad9. Rad17-RFC binds to nicked circular, gapped, and primed DNA and recruits the 9-1-1 complex in an ATP-dependent manner. Electron microscopic analyses of the reaction products indicate that the 9-1-1 ring is clamped around the DNA.

We report magnetic susceptibility measurements on a layered superconductor Li0.48(THF)0.3HfNCl having Tc approximately 26 K. The present study revealed that (a) the Fermi level density of states is small, N*(EF) approximately 0.25 states/(eV spin f.u.), (b) mass enhancement is negligible, gamma; approximately 1, (c) electron-phonon coupling is weak, lambda(ep)<<1, (d) exchange enhancement is negligible, 1/(1+F(a)0) approximately 1, and (e) electronic density parameter is large, r(2D)s approximately 10.3 (i.e., low-carrier density). It is difficult to explain the origin of the high Tc in terms of the conventional phonon (BCS) mechanism of superconductivity.

Laser ablation under bronchoscopic guidance was conducted on 2 patients with severe tracheal stenosis. Case 1 was a 57-year-old man admitted to our emergency unit because asphyxia. Bronchoscopy showed the lumen occluded at the bifurcation by tracheal cancer. Case 2 was a 62-year-old woman who underwent tracheostomy elsewhere for respiratory failure caused by a brain contusion and was treated for 3 months. After transfer to our emergency unit, bronchoscopy showed severe tracheal stenosis. Tracheoplasty conducted under bronchoscopy used a noncontact Nd:YAG laser at an output of 10-40 W and irradiation time of 1 second per shot. Total irradiation energy was 1700-1900 J. Percutaneous cardiopulmonary support was used during the laser procedure due to asphyxia. All procedures were completed satisfactorily and clinical symptoms improved dramatically in both cases. Laser tracheoplasty under bronchoscopic guidance treated severe stenosis safely and completely. Percutaneous cardiopulmonary support was very useful in preventing severe respiratory failure or asphyxia during this procedure.

We describe herein our new technique of performing video-assisted thoracic surgery (VATS) using a flexible scope and a bleb implement which allows for a reduction in the risk of overlooking small and flat blebs in patients with a spontaneous pneumothorax (SP). This technique was performed in 26 consecutive patients with a SP. A skin incision about 18 mm long was made in the area over which the patients had specified preoperatively, and meticulous observation of the apex and superior segment was performed using a bleb implement. Partial pneumonectomy with endoscopic staplers and laser ablation were carried out for pleural lesions. We found that VATS combined with a flexible scope and a bleb implement resulted in better visibility for detection and removal of any small and flat blebs.

The number of mutant p53 protein-positive tumor cells in primary non-small cell lung carcinoma (NSCLC) cases were quantitated by flow cytometry (FCM) and the relationships of these data to various factors were evaluated. Furthermore, the method of quantitating telomerase activity was investigated. Forty patients with primary lung carcinoma encountered between December 1995 and December 1997 were investigated. Among these cases, telomerase activity was measured. Using PAb421, cells were reacted with fluorescent antibody and fluorescence was quantitated by FCM. Fluorescence index (FI) was estimated in relation to the positivity rates of negative controls and were quantitatively evaluated. FI values of normal lung tissue were obtained from normal lung tissue excised from young patients with pulmonary bulla. Values that were 2SD or more above the mean value of normal lung tissue (> 2.19) were regarded as mutant p53-positive, and 14 (35.0%) of 40 lung carcinoma cases were positive by this criterion. Of 13 poorly differentiated carcinoma cases, seven cases (53.8%) were positive, which was significantly high. Furthermore, the telomerase activity was converted to numerical values in 40 cases using the telomelic repeat amplication protocol (TRAP) method as well as the total product generated (TPG) method. The mean TPG value of the 40 cases was 75.21 +/- 15.63. Among these cases, the mean value of fourteen p53-positive cases was 124.49 +/- 37.19, which was higher than that of 26 negative cases, 48.68 +/- 10.88, showing a significant difference. The method used in this study was considered a useful method that allows accurate and objective evaluation of mutant p53 expression. It was suggested that mutant p53 expression may affect the degree of tumor cell differentiation. Consequently, it was confirmed in this study that mutant p53 expression and telomerase activity were closely associated in lung cancer.

Objective: To investigate the role of pleural lavage cytology (PLC) in resection for primary lung carcinoma., Summary Background Data: The prognostic significance of PLC before manipulation is still controversial., Methods: Cytology of pleural lavage immediately after thoracotomy but before any manipulation of the lung was examined in 500 consecutive patients with lung cancer with no pleural effusion who underwent pulmonary resections. Eighteen patients who already had pleural dissemination were excluded from this study., Results: Eighteen of 482 patients (3.7%) had positive cytologic findings. The positivity of PLC was significantly correlated with histology, extension of tumor to pleura, and presence of lymphatic permeation or vascular involvement by tumor. Positive lavage findings were seen only in adenocarcinoma. Because 6.3% of the patients with adenocarcinoma had positive cytologic findings, it is vital to perform PLC before curative resections for lung cancer, especially adenocarcinoma. The 5-year survival rates of the patients having negative and positive lavage findings were 52.9% and 14.6%, respectively. The prognosis of the patients with positive lavage findings was as poor as that of the patients with stage IIIB disease and that of the patients with malignant effusion., Conclusions: Positive findings on PLC indicate exfoliation of cancer cells into the pleural cavity, which is an essential prognostic factor. In addition, we should regard positive cytologic findings as a subclinical malignant pleural effusion that is pathologic stage T4.

We report the use of video-assisted thoracic surgery (VATS) as a treatment for pulmonary metastases. Eight patients with metastatic lung cancer were treated with VATS techniques. These patients included 5 males and 3 females whose ages ranged from 34 to 61 years (average: 47.4). Their primary diseases were seminoma (n = 3), renal cell carcinoma (n = 2), colon carcinoma (n = 1), mammary carcinoma (n = 1) and choriocarcinoma (n = 1). Computed tomography (CT) scans with current generation scanners were performed preoperatively and revealed one metastatic lesion in each of five cases and two lesions in each of the other three cases. In one case, one lesion was located in each of the lungs. Tumor size ranged from 5 to 30 mm in diameter, and all lesions were in the peripheral field of the lung. VATS was carried out with three surgical ports for three cases, two surgical ports for one case, and only one port for two cases. For all cases, an endo-stapler was utilized. In two cases, the preoperative point-marking technique for tumors was employed under the guidance of CT imaging. All patients were discharged from the hospital with no complications, and were followed up with no evidence of lung recurrence. Our criteria in selecting patients for the VATS removal of metastatic lung tumors are as follows: 1) tumors are less than 30 mm in diameter in the peripheral lung, 2) the number of tumors detected by CT scan, etc., is one or two. We conclude that VATS is a good candidate for the resection of lung metastases in the selected cases. Long term outcome remains to be solved in the future.

Background: The use of surgery for metastatic lung cancer has been established recently and the indications have been extended to multiple and bilateral lung metastases. However, in some patients, secondary lung metastasis appears soon after the first pulmonary surgery, making curative treatment very difficult. Postoperative weakness of tumor angiogenesis suppression mechanisms seems to play an important role in the recurrence of lung metastases. To verify this hypothesis, we performed a clinical and an experimental study., Results and Conclusion: The clinical study revealed that serum vascular endothelial growth factor (VEGF), also known as vascular permeability factor, increased after pulmonary surgery. The experimental study showed that VEGF played an important role in the rapid growth of dormant micrometastases of the lung. These results suggested that the postoperative increase of VEGF disrupted angiogenesis suppression and induced the growth of dormant micrometastases early in the postoperative period. It was also demonstrated that this effect of VEGF on micrometastases was abolished by AGM-1470, an angiogenesis inhibitor. In conclusion, postoperative treatment with AGM-1470 might inhibit the early recurrence of malignant tumors.

Surgical treatment for metastatic lung tumor has also been aggressively performed to treat multiple or bilateral lesions recently. However, in some patients, metastatic pulmonary foci have recurred after surgery for metastatic lung tumor. These foci could not be controlled even after performing thoracotomy several times in some patients. In this study, we examined prognostic factors in patients undergoing surgery for metastatic lung tumor with respect to early relapse of metastatic pulmonary foci after surgery, and discussed strategies for improving long-term results. This study included 120 patients who underwent surgery for metastatic lung tumor in our department between November 1975 and November 1997. Overall, the 5-year survival rate was 37.1%. When results were compared among groups, there were no significant differences related to age, gender, primary organ, DFI, number of metastatic foci or surgical technique. However, the prognosis was significantly poorer in patients with recurrent metastatic pulmonary foci after surgery. Especially in patients with early relapse within 6 months after resection of the lung, the prognosis was markedly poor. Early relapse was an important factor involved in unfavorable surgical outcomes. The mechanism involved in the early relapse of metastatic pulmonary foci after surgery for metastatic lung tumor may be associated with the presence of several micrometastases that could not be recognized during surgery for metastatic lung tumor, that is, dormancy, in the lung. Surgical outcomes in patients with metastatic lung tumor will be improved if a method of controlling this increase in dormant metastases is established.

Surgical intervention is routinely employed for non-small cell lung cancer, whenever no distant metastasis is found. However, its surgical results depend on the staging of the lung cancer, and surgery for stage III disease is less effective compared with stage I or II disease. For patients with stage III, some treatments such as induction chemotherapy have been tried to support the surgery. Recently, T3 lung cancer is divided in two category of staging, T3N0 is evaluated to be stage II B and T3N1-2 is stage III A. In this study, we classified T3 diseases by some factors, and examined long-term survival of each group. As a result, we concluded that mediastinal invasion and N2 involvement were risk factors to early failure. On the contrary, patients with adjacent lobe invasion and resection of two lobes led to the prolonged survival rather than lobectomy with partial resection of adjacent lobe.

Home medical care is becoming a greater matter of concern along with the increasing population of elderly persons. The various difficulties in home care are also found in regional care, and it could be said that in Japan regional care serves as an "advanced model" for home medical care. Ohya Town is located in the mountains of northern Hyogo Prefecture. It has a population of about 5,000 of which 32% is elderly people. There is no railroad, and it takes around 30 minutes to travel to the central hospital from the area. The three town clinics are in contact with each other and with the central hospital concerning medical information. The characteristics of Home Parenteral Nutrition in this underpopulated district are discussed and reported in reference mainly to cases in which the 3 way valved peripherally inserted central venous catheter (Groshong Catheter) is used, in comparison with HPN cases in the city university hospital.

We report a case of operation against chronic expanding hematoma developed in the pleural cavity of the patient who were treated as anaplastic anemia. 69-year-old man had been underwent thymectomy because of the thymoma and hemogenic pleural effusion 8 years ago. Then, last year, he was pointed out severe thrombocytopenia and underwent transfusion of thrombocyte. In the same period, chest X-ray examination revealed development of the tumor-like shadow in the left lung field. The abnormal shadow has enlarged, so the patient admitted us. X-ray findings revealed the huge tumor occupied completely the left pleural space. He was also pointed out severe pancytopenia (WBC: 2,600/mm3, RBC: 172 x 10(4)/mm3, thrombocyto: 1.9 x 10(4)/mm#), and the examination of bone marrow revealed anaplastic anemia. Thoracotomy was underwent, and the tumor was resected. Pathological diagnosis of the tumor was huge hematoma. Negative pressure in the pleural space would keep the hemorrhage and make hematoma huge. The hematoma in the pleural space should be resected as soon as the diagnosis is decided.

Airway obstruction due to oppression by the right aortic arch is relatively common in the newborn, but has never been reported in the adult. We reported an aged case of right main bronchial stenosis due to the oppression by the right aortic arch, performed successful bronchodilatation using the stent. A 92-year-old male complained of fever and productive cough and hospitalized with a diagnosis of pneumonia. Close examinations revealed that his right main bronchus was stenotic due to oppression by the right aortic arch. Bronchodilatation was performed successfully using a Gianuturco-type expandable metallic stent. No endoluminal stenosis was found during 12 months follow up. For the treatment of bronchial deformity by the extrabronchial oppression, implantation of the Gianturco-type expandable metallic stent is considered to be very useful method.

A 46-year-old woman complained of a back pain and was pointed the abnormal shadow on chest X-ray by the physician, so she consulted our hospital in January 1987. After a few weeks, the size of the shadow decreased with no treatment, then we followed up her with roentgenograms. Since '91, its size had increased again. Needle biopsy did not give us the diagnosis, but we performed the operation because we suspected some malignant disease. We resected the tumor with the enough surgical margin. Its size was 4 x 3 x 3 cm, its surface was white and smooth. Its was elastic hard and had the pedicle which jointed the visceral pleura. Histological diagnosis was the localized fibrous mesothelioma.

We report herein the case of a patient in whom a calcified tuberculous empyema with multiple fistulae was successfully treated by a new surgical approach. Reexpansion of the affected lung which had calcified over 30 years was achieved by covering the multiple bronchial fistulae using the omentum without obliterating the empyema cavity. Although the patient presented with severe aspiration pneumonia, he made a complete recovery and is now leading a better quality of life than before. This new operative method is less invasive and can therefore be performed much more easily on critically ill patients than conventional methods.

To evaluate the effects of ceftriaxone (CTRX) administered once daily, the biliary concentration of CTRX and daily changes in the bacterial count in bile from an indwelling T-tube were measured concomitantly. The effects on prophylaxis of postoperative infections after biliary tract surgery were also examined mainly due to clinical symptoms. The biliary CTRX levels increased rapidly and were sustained as high as 92.5-219 micrograms/ml in all patients even 24 hours after 2 g of CTRX was infused intravenously once daily. Five of 7 patients whose bile samples were positive for bacteria showed high CTRX levels in bile and almost no bacteria present after treatment. CTRX-sensitive bacteria eliminated 1 or 2 hours after the administration in response to the increase of biliary CTRX levels. The changes in bacteria count of bile may be closely related to CTRX antibiotic activity. As an antibiotic prophylaxis, other 18 patients with biliary diseases received intravenous infusion of 2 g of CTRX once daily for 5-7 days (mean 5.9 days). Thus, once-daily dose treatment of CTRX 2 g may have antibiotic effects on biliary infection and postoperative prophylaxis of biliary infections and it is indicated that the prolonged biliary levels of CTRX are essential for its efficacy.