Genetic features of pulmonary adenocarcinoma presenting with ground-glass nodules: the differences between nodules with and without growth.

BACKGROUND: Pulmonary ground-glass nodules (GGNs) include both malignant and benign lesions. Some GGNs become larger, whereas others remain unchanged for years. We have previously reported that smoking history and large diameters are predictors for growth. However, the genetic differences among GGNs remain unclear. PATIENTS AND METHODS: GGNs with ground-glass component of ≥50% on a thin-section computed tomography scan that were resected between 2012 and 2014 were evaluated for clinicopathological features and the presence of EGFR/KRAS/ALK/HER2 mutations. 'Incidence of 2-mm growth' and 'Time to 2-mm growth' were analyzed according to the mutational status. RESULTS: Among 104 GGNs in 96 patients, this study included 3 atypical adenomatous hyperplasia (AAH), 19 adenocarcinoma in situ (AIS), 27 minimally invasive adenocarcinoma (MIA), and 55 invasive adenocarcinoma (IA). Among the 71 lesions evaluable for growth, 30 GGNs exhibited growth and 5 lesions remained unchanged for ≥2 years before surgery was carried out. We identified mutations or rearrangements in 75% of GGNs (78/104). EGFR mutations were noted in 64% of samples, KRAS in 4%, ALK in 3%, and HER2 in 4%. The remaining 26 quadruple-negative tumors were significantly associated with AAH/AIS (P < 0.01) and no-growth (P < 0.01) compared with driver mutation-positive tumors, whereas EGFR mutation-positive tumors were correlated with MIA/IA (P < 0.01) and growth (P < 0.01) compared with EGFR-negative tumors. CONCLUSIONS: Three fourths of resected GGNs were positive for EGFR, KRAS, ALK, or HER2 mutations. Quadruple-negative tumors were associated with a lack of GGN growth, whereas EGFR mutation-positive tumors displayed a correlation with growth.

Subtypes of peripheral adenocarcinoma of the lung: differentiation by thin-section CT.

The purpose of this study was to scrutinize morphological characteristics of thin-section CT of the histopathological subtypes of adenocarcinoma of the lung. The subjects consisted of 83 patients with 87 adenocarcinomas measuring 3 cm or less in the largest. The tumors were divided into three groups (group I: Noguchi's histological subtypes type A and B tumors, group II: type C tumors, and group III: type D, E, and F tumors). In each group, tumor size, shape (round versus polygonal), presence of air bronchogram, bubble-like areas, coarse spiculation, pleural tag, and ratio of ground glass attenuation (GGA) were evaluated. Most of the group II lesions showed polygonal shape, whereas tumors in other groups were round in shape (P<0.01). Air bronchogram and bubble-like areas of low attenuation was seen more frequently in group II compared with those in group III (P<0.01). GGA areas were largest in group I and smallest in group III (P<0.01). We believe thin-section CT findings reflect the histopathological subtypes of adenocarcinoma of the lung. The presence of air bronchogram and bubble-like areas of low attenuation areas in particular is useful to differentiate replacement growth tumors from non-replacement growth tumors.

Hybridization of oligonucleotide by using DNA self-assembled monolayer.

The interaction between DNA immobilized on surface and oligonucleotides at the interface is important in detection and diagnostic processes. However, it is difficult to immobilize DNA with maintaining its activity and to realize an efficient hybridization in previous methods. Here, to establish a novel DNA-functionalized surface, the DNA self-assembled monolayer (SAM) was constructed on a gold substrate using thiolated DNA composed of double-stranded (ds) and single-stranded (ss) portion. The DNA SAM was characterized by surface plasmon resonance (SPR), XPS. The hybridization of ss portion of DNA was attempted using the SAM, and in situ monitored by SPR. XPS measurement indicated that the thiolated DNA could form a stable monolayer on a gold substrate through sulfur-gold interaction. SPR measurement implied that the long axis of the DNA standing on the substrate. These results indicated formation of the DNA SAM on the substrate. Hybridization of target DNA containing a complementary sequence for the probe portion was observed by SPR. Moreover, one mismatch of oligonucleotide could be distinguished using the DNA SAM. The SPR result indicates that hybridization of target DNA and probe DNA on the DNA SAM occurs on the DNA SAM.

Association of IL-8 and MCP-1 with the development of reexpansion pulmonary edema in rabbits.

BACKGROUND: The aim of this study is to determine the relationships between the cytokines and the inflammatory response in reexpansion pulmonary edema (RPE). METHODS: We examined the cell population, epithelial permeability measured by Evans blue dye (EB), betaglucuronidase and cytokine concentrations in bronchoalveolar lavage fluid (BALF) and/or blood using a rabbit RPE model. RESULTS: We confirmed that RPE is characterized by recruitment of polymorphonuclear leukocytes (PMNs), the release of PMN granular contents into the air spaces, and increased vascular permeability. These findings were highly correlated with increased interleukin-8 (IL-8) and monocyte chemoattractant protein 1 (MCP-1) concentrations in the BALF. Growth related oncogene (GRO) was detected in the BALF from only 2 of the 7 reexpanded lungs while TNFalpha was not detected in any rabbits. A similar but less severe inflammatory response to the reexpanded lung was found in the contralateral lung. CONCLUSIONS: IL-8 and MCP-1 may play important roles in the development of RPE; the inflammatory response is independent of TNFalpha and unilateral reexpansion of the lung induces an inflammatory response not only in the reexpanded lung but also in the contralateral lung.

Mouse proteasomal ATPases Psmc3 and Psmc4: genomic organization and gene targeting.

PSMC3 and PSMC4, components of the 19S complex of the 26S proteasome, show a significant degree of amino acid similarity, especially in the conserved ATPase domain (CAD). In this study, we characterized the mouse Psmc3 and Psmc4 genes. The genomic structures of both genes showed a significant degree of similarity. The Psmc3 gene was composed of 12 coding exons, whereas the Psmc4 gene had 11 exons. Exons encoding the leucine zipper domain and CAD were identical in number between the Psmc3 and Psmc4 genes. The Psmc3 gene mapped to mouse chromosome 2, whereas Psmc4 mapped to chromosome 7. We further addressed the biological roles of Psmc3 and Psmc4 through the generation of gene targeted mice. Both Psmc3- and Psmc4-deficient mice died before implantation, displaying defective blastocyst development. These findings indicate that Psmc3 and Psmc4 have similar and essential roles in early embryogenesis and further that both ATPases have noncompensatory functions in vivo.

Cutting edge: endotoxin tolerance in mouse peritoneal macrophages correlates with down-regulation of surface toll-like receptor 4 expression.

Monocytes/macrophages exposed to LPS show reduced responses to second stimulation with LPS, which is termed LPS tolerance. In this study, we investigated molecular mechanism of LPS tolerance in macrophages. Mouse peritoneal macrophages pre-exposed to LPS exhibited reduced production of inflammatory cytokines in a time- and dose-dependent manner. Activation of neither IL-1 receptor-associated kinase nor NF-kappaB was observed in macrophages that became tolerant by LPS pretreatment, indicating that the proximal event in Toll-like receptor 4 (TLR4)-MyD88-dependent signaling is affected in tolerant macrophages. Although TLR4 mRNA expression significantly decreased within a few hours of LPS pretreatment and returned to the original level at 24 h, the surface TLR4 expression began to decrease within 1 h, with a gradual decrease after that, and remained suppressed over 24 h. A decrease in inflammatory cytokine production in tolerant macrophages well correlates with down-regulation of the surface TLR4 expression, which may explain one of the mechanisms for LPS tolerance.

alpha-Chemokine growth factors for adenocarcinomas; a synthetic peptide inhibitor for alpha-chemokines inhibits the growth of adenocarcinoma cell lines.

PURPOSE: The experiments aimed to determine if alpha-chemokine inhibitors are effective suppressors of the growth of adenocarcinomas, a neoplasm with a high mortality rate. METHODS: Expression of growth-related oncogene (GROalpha) and interleukin-8 (IL-8) was determined by enzyme-linked immunosorbent assay. Inhibition of alpha-chemokine binding to tumor cells was assessed in the presence and absence of the hexapeptide, antileukinate. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assays were performed to determine the effect of alpha-chemokines, monoclonal antibodies (mAb), and antileukinate on cell proliferation. Finally, antileukinate inhibition of human, lung adenocarcinoma tumor growth, was determined in BALB/c nude mice. RESULTS: All of the adenocarcinomas tested produced either GROalpha or IL-8 or both. Proliferation of lung, stomach and colon adenocarcinoma cells was inhibited by anti-GROalpha mAb and/or anti-IL-8 mAb while recombinant human GROalpha stimulated the proliferation of lung and stomach adenocarcinomas. Antileukinate inhibited GROalpha binding to specific receptors on adenocarcinoma cells and inhibited the proliferation of all adenocarcinomas tested. Colon-derived adenocarcinomas specifically bound IL-8 and this binding was also inhibited by antileukinate. Administration of antileukinate in vivo inhibited the tumor growth of adenocarcinoma A549. CONCLUSIONS: GROalpha and IL-8 are necessary for the growth of lung, stomach and colon adenocarcinomas, and can be inhibited by the hexapeptide, antileukinate. The findings suggest the possibility of using alpha-chemokine receptor inhibitors in the treatment of adenocarcinoma.

Cathepsin Y (a novel thiol enzyme) produces kinin potentiating peptide from the component protein of rat plasma.

Rat spleen cathepsin Y (a novel enzyme) that produces bradykinin (BK) potentiating peptide (BPP) from rat plasma was isolated, characterized and its amino acid sequence was deduced from cDNA cloned by reverse transcription-polymerase chain reaction (RT-PCR). We propose the name cathepsin Y for this enzyme considering its origin, characteristics and the amino acid sequence. BPP potentiates not only BK but also lysyl-BK (lysBK) and T-kinin (TK) action on uterus contraction. The structure of BPP is Pro-Pro-Pro-Leu-Gly-Pro-Gly-Ser. The magnitude of the potentiation of BK activity by synthesized BPP was seven-fold when equivalent quantities added to BK and 23-fold when the level is doubled. The precursor proteins that produce BPP by the action of cathepsin Y are eluted into two fractions when the heated plasma was applied to a negative ion exchange column. Structure relationships between these two proteins are now under investigation. In this paper, we report on the characteristics and the amino acid sequence of rat spleen cathepsin Y, its structure and the potentiating activity of BPP, and isolation of the precursor protein.

IL-18-deficient mice are resistant to endotoxin-induced liver injury but highly susceptible to endotoxin shock.

IL-18 is an IL-1-related cytokine which shares biological functions with IL-12. These include the activation of NK cells, induction of IFN-gamma production and Th1 cell differentiation. In this study we analyzed the effect of IL-18 deficiency on lipopolysaccharide (LPS)-induced liver injury and endotoxin shock in Propionibacterium acnes-primed mice. P. acnes-primed IL-18-deficient (IL-18KO) mice showed resistance to LPS-induced liver injury. Unexpectedly, P. acnes-primed IL-18KO mice were highly susceptible to LPS-induced endotoxin shock. Serum level of tumor necrosis factor (TNF)-alpha were markedly elevated (approximately 10-fold higher) within 1.5 h after LPS challenge in IL-18KO mice as compared with wild-type mice. Anti-TNF-alpha antibody administration to IL-18KO mice was significantly protective against endotoxin-induced lethality. P. acnes-primed IL-18KO macrophages produced approximately 6-fold more TNF-alpha protein than did P. acnes-primed wild-type control macrophages. Taken together, these findings demonstrate that IL-18 is responsible for the progression of endotoxin-induced liver injury as well as down-regulation of endotoxin-induced TNF-alpha production in P. acnes-primed mice.

Microsatellite instability and frameshift mutations in the Bax gene in hereditary nonpolyposis colorectal carcinoma.

We studied microsatellite instability (MI) and bax gene abnormalities in colorectal carcinomas from 36 patients diagnosed as having hereditary nonpolyposis colorectal cancers (HNPCC) according to the clinical criteria (12 with confirmed HNPCC in group A and 24 at high risk of HNPCC in group B) and from 20 randomly selected patients with other colorectal cancers. MI was examined at 4 dinucleotide microsatellite loci and one mononucleotide locus. Frameshift mutations in the bax gene were detected by polymerase chain reaction-single strand conformation polymorphism analysis. MI was detected in 7 of the 12 patients in group A and 12 of the 24 in group B. Three MI patterns were identified: type 1, MI in both mono- and dinucleotide repeats; type 2, MI only in mononucleotide repeats and type 3, MI only in dinucleotide repeats. Most MI-positive patients in group A showed type 1 MI, whereas in group B, 5 showed type 1, 3 showed type 2 and 4 showed type 3. Frameshift mutations in the bax gene correlated strongly with type 1 and type 2 MI. These results indicate that mutations of different DNA mismatch repair genes may cause several types of MI and result in several different clinical phenotypes of HNPCC. The bax gene may be one of the target genes which play a role in the tumorigenesis of HNPCC.

Excision of a recurrent pericardial cyst using video-assisted thoracic surgery.

A recurrent pericardial cyst manifested as an enlarging cardiophrenic cyst and was reexcised with the aid of video-assisted thoracic surgery. Video-assisted thoracic surgical excision can be a safe and effective approach, but complete excision should be performed to avoid recurrence.

Inducible expression of nuclear factor IL-6 increases endogenous gene expression of macrophage inflammatory protein-1 alpha, osteopontin and CD14 in a monocytic leukemia cell line.

Nuclear factor-IL-6 (NF-IL6) belongs to the CCAAT/enhancer binding protein family of transcription factors. NF-IL6 binds to the regulatory regions of many genes induced in activated macrophages in vitro. However, which particular genes are regulated by NF-IL6 in vivo is poorly defined. In order to identify the downstream genes of NF-IL6 in a monocytic lineage, we combined an inducible expression system with subtraction cloning in a study of murine M1 monocytic leukemia cells. We demonstrated that inducible expression of NF-IL6 is able to increase endogenous gene expression of macrophage inflammatory protein (MIP)-1 alpha, osteopontin and CD14 in M1 cells. We also showed that NF-IL6 activated murine MIP-1 alpha proximal promoter luciferase construct which contains two NF-IL6 binding sites and a point mutation of either site markedly reduces the luciferase activity. These findings indicate that MIP-1 alpha is a direct target of NF-IL6.

[Prognosis and prognostic factor after extended lymphadenectomy in lung cancer].

From 1984 to 1994, 418 patients were received surgery for the lung cancer in our center. Of them, 178 patients were underwent extended lymphadenectomy. Fifty six of the 178 were histologically proven N2 of N3 alpha disease after surgery. Extended lymphadenectomy means R2b lymphadenectomy including the left tracheobronchial node dissection for the right lung cancer and R3 (bilateral mediastinal lymphadenectomy through a median sternotomy) for the left. In the 56 patients, we examined the location and frequency of metastases to the mediastinal lymph nodes and the relationship between some clinical factors (pT, number of metastatic station, clinical staging of the lymph node (CN), histological type, contralateral mediastinal lymph nodes metastases) and prognosis. Most of the pN2 patients of the right lung cancer showed ipsilateral mediastinal lymph nodes metastases and 25 percent of the patients showed the spread to N2b mediastinal modes. The patients of the left lung cancer showed higher incidence of contralateral mediastinal lymph nodes metastases than the patients of the right lung cancer. The five years survival rate of all pN2 patients (N = 39) was 48%, and T1 or T2-N2 patients (N = 22) was 67%. On the other hand, all T4 N2 patients (N-9) died within 3 years after operation. There was no significant difference in postoperative survival between the patients with single station metastasis (N = 20) and multistation metastases (N = 30, including 11 cases with N3 alpha). The five years survival rate of all the patients with multistation metastases was 45%, and that of T1 or T2 multilevel (N = 20) was 65%. There was significant difference in postoperative survival between the patients with CN0-pN2 and CN2-pN2 (p < 0.05). The five years survival of CN0-pN2 patients (N = 14) was 85% and four years survival rate of CN2 pN2 patients (N = 19) was 30%. Among the patients with T1 or T2 tumor, however, there was no significant differences in postoperative survival between CN0-pN2 patients and CN2 pN2 patients. There was no difference in postoperative survival between adenocarcinoma and squamous cell carcinoma (5 years survival: 56%, 43%). In conclusion, extended lymphadenectomy has brought a good prognosis in the patient with T1 or T2 in spite of presence of CN2 or multistation N2. The patients with contralated mediastinal metastases (N3 alpha) showed good prognosis after R3 (5 years survival: 100%) in the patients with the left lung cancer (N = 6). But the N3 alpha patients of right lung cancer showed poorer prognosis (3 years survival: 30%) after R3 (N = 5) than the left. It suggested that R3 lymphadenectomy was significant and beneficial for the left lung cancer patients with N3 alpha but controversial for the right.

[An approach to lobectomy under a thoracoscopic surgery].

Lobectomy under a thoracoscope is a minimally invasive surgery and full consideration must also be given to the length of access thoracotomy and the number of access ports inserted. On the other hand, securing a full visual field within a limited access and treating the pulmonary artery/vein and bronchus safely and for sure are needed. However, it would not be too much to say that the easiness of lobectomy under a thoracoscope depends on the length of access thoracotomy and the number of access ports inserted. In the experiment this time we discussed an approach to the access area in lobectomy under a thoracoscope in our cases.

Primary solitary amyloidoma of the lung: findings on CT and MRI.

Pulmonary amyloidoma is a rare disease which is usually found incidentally on chest radiographs in asymptomatic, elderly people. Amyloid nodules may be solitary or much more commonly multiple. There have been many reports of radiological findings of pulmonary amyloidosis; however, those have not been characteristic. We report the findings on CT and MRI of a proven primary pulmonary amyloidoma in an asymptomatic 76-year-old woman. The low intensity of the lesion on T2-weighted images may be useful in the differential diagnosis from bronchogenic carcinoma.

[Investigation into mediastinal lymph node metastasis of lung cancer and rationale for decision of the extent of mediastinal dissection].

UNLABELLED: From the study on the regional lymphatic drainage and investigation into mediastinal lymph node metastasis of lung cancer, we have decided the extent of mediastinal dissection as follows; 1) For right lung cancer, as the routine procedure, extended systematic ipsilateral mediastinal dissection including the left tracheobronchial region, the anterior and the posterior ipsilateral mediastinum through a conventional thoracotomy. 2) For left lung cancer, as the routine procedure, systematic bilateral mediastinal dissection through a median sternotomy. 3) For the patients with advanced lymph node metastasis (the highest mediastinal or the cervical node involvement) or direct extension into the upper mediastinum of cancer in any side of the lungs, the lower half of modified radical neck dissection combined with systematic bilateral mediastinal dissection through a cervical collar incision and median sternotomy. RESULTS: 1. The noteworthy location and incidences of mediastinal lymph node involvement were as follows; 1) Among 34 patients of right lung cancer with pN2-3 M0 disease, in 5 patients the anterior mediastinal node involvement and in 6 patients (18%) the contralateral tracheobronchial node involvement were found by the pathological investigation at surgery. 2) The incidences of contralateral mediastinal node involvement at median sternotomies were 20% of 15 patients of the left upper lobe primary and 57% of 7 patients of the left lower lobe primary. 2. Postoperative survival rates calculated with Kaplan-Meier method; 1) The five-year survival rates were 67% in 22 patients with pT1-2N2M0; 72% in 20 patients with pT1-2N2-3 alpha (one level) M0 and 65% in 13 patients with pT1-2N2-3 alpha (multi level) M0. 2) The five-year survival rate of 8 patients with N3 gamma whose cancer were diagnosed as cN0-3 alpha preoperatively and resected completely was 60%. In conclusion, these results encourage us to continue this study because we can believe that our systematic mediastinal dissection beyond the anatomical difficulties would bring better prognoses in the patients with pN2-3 disease.

Ectopic expression of CHOP (GADD153) induces apoptosis in M1 myeloblastic leukemia cells.

CHOP (GADD153) is a member of the C/EBP family and a stress-induced protein. To investigate the role of CHOP in cellular growth, we expressed CHOP conditionally in M1 myeloblastic leukemia cells that do not express p53 protein. More than 60% of M1 cells died through apoptosis 72 h after CHOP induction. Site-directed mutagenesis revealed that this process requires leucine zipper domain but neither intact basic region nor trans-activation domain. CHOP-mediated apoptosis accompanied downregulation of bcl-2 mRNA and overexpression of Bcl-2 delayed the process. Our results indicate that CHOP can induce apoptosis in a p53-independent manner.

[Evaluation of neck lymph node dissection and extended lymphadenectomy through a collar incision and median sternotomy for lung cancer].

Since 1983, 421 patients have been treated for lung cancer at this institute. Since 1988, neck lymph node dissection (11 cases) and new extended lymphadenectomy through a collar incision and median sternotomy (22 cases) have been conducted. Indications for this new radical operation are scalene, supraclavicular or highest mediastinal node involvement, or superior pulmonary sulcus carcinoma, in patients aged 70 or less without distant metastasis and NSCLC. No major complications and operative mortality were encountered in this study. Patients with scalene or supraclavicular node involvement showed poor prognosis. Postsurgical local recurrence was frequent. Whether resection in N3 disease should be conducted or not, remains a point of controversy. The authors consider that lymphadenectomy should be conducted more extensively. A significant better survival of N2 disease and satisfactory prognosis of patients without metastasis of cervical lymph nodes demonstrates the effectiveness of neck lymph node dissection in the present superradical operation for lung cancer.