Oral squamous cell carcinoma with microscopic extracapsular spread in the cervical lymph nodes.

The purpose of this study was to determine the detailed background of cases of oral squamous cell carcinoma (OSCC) with microscopic extracapsular spread (ECS) in the cervical lymph nodes. The cases of 78 patients with primary OSCC, who attended hospital from October 2007 to July 2011 and underwent resection of the primary tumour with neck dissection, were reviewed. The subjects were classified into three categories: pN0, pN+/ECS-, and pN+/ECS+; the outcomes of pN+/ECS+ patients were compared in detail with those of the other categories. Thirty-one cases (39.7%) were pN0, 25 cases (32.1%) were pN+/ECS-, and 22 cases (28.2%) were pN+/ECS+. The 3-year overall survival rate was 82.1% in pN0, 74.1% in pN+/ECS-, and 39.8% in pN+/ECS+ (pN0 vs. pN+/ECS+, P=0.0004; pN+/ECS- vs. pN+/ECS+, P=0.0086). The 3-year disease-specific survival rate was 96.2% in pN0, 77.2% in pN+/ECS-, and 39.8% in pN+/ECS+ (pN0 vs. pN+/ECS+, P<0.0001; pN+/ECS- vs. pN+/ECS+, P=0.0038). Patients with poorly differentiated carcinoma, those with three or more ECS+ nodes, and those with ECS+ node(s) located at levels III, IV, and V, had the worst prognosis among pN+/ECS+ subjects.

[Resection of a right ventricular diverticulum of fibrous type by off-pump cardiac surgery; report of a case].

Right ventricular diverticulum is very rare and we experienced a case of isolated right ventricular diverticulum in an adult patient The patient was an 80-year-old man and a 3-cm-diameter round mass at the apex of the heart was pointed out by screening computed tomography (CT). A small and akinetic diverticulum having a narrow communication with the right ventricle was revealed by right ventriculography. Upon surgery, a 3-cm-diameter diverticulum was found at the acute margin of the right ventricle. The diverticulum was exposed using the off-pump coronary artery bypass grafting (CABG) technique. Two mattress sutures of 4-0 Prolene with Teflon felt strips were used to close the communication between the diverticulum and the right ventricle, then, the diverticulum was resected. His postoperative course was uneventful. Pathological examination revealed the endothelial-lined wall of the diverticulum consisting of internal elastic lamina and discontinuous thin smooth muscle layer with no myocardium. This type of right ventricular diverticulum could be resected by the off-pump CABG technique.

Induction of apoptosis and cellular senescence in mice lacking transcription elongation factor, Elongin A.

Elongin A is a transcription elongation factor that increases the overall rate of mRNA chain elongation by RNA polymerase II. To gain more insight into the physiological functions of Elongin A, we generated Elongin A-deficient mice. Elongin A homozygous mutant (Elongin A(-/-)) embryos demonstrated a severely retarded development and died at between days 10.5 and 12.5 of gestation, most likely due to extensive apoptosis. Moreover, mouse embryonic fibroblasts (MEFs) derived from Elongin A(-/-) embryos exhibited not only increased apoptosis but also senescence-like growth defects accompanied by the activation of p38 MAPK and p53. Knockdown of Elongin A in MEFs by RNA interference also dramatically induced the senescent phenotype. A study using inhibitors of p38 MAPK and p53 and the generation of Elongin A-deficient mice with p53-null background suggests that both the p38 MAPK and p53 pathways are responsible for the induction of senescence-like phenotypes, whereas additional signaling pathways appear to be involved in the mediation of apoptosis in Elongin A(-/-) cells. Taken together, our results suggest that Elongin A is required for the transcription of genes essential for early embryonic development and downregulation of its activity is tightly associated with cellular senescence.

A ubiquitin ligase, skeletrophin, is a negative regulator of melanoma invasion.

Skeletrophin (mindbomb homolog 2 (MIB2)) is a RING (Really Interesting New Gene) finger-dependent ubiquitin ligase, which targets the intracellular region of Notch ligands. A previous immunohistochemical study demonstrated that skeletrophin was downregulated in many melanomas. In the present study, we have identified a promoter region of skeletrophin on a CpG island and detected aberrant methylation of this region in six of 31 invasive melanomas, but in none of 25 benign nevi or five non-invasive superficial spreading melanomas. Subsequently, we found that a zinc-finger transcriptional factor Snail, which is overexpressed in many melanoma cells, repressed the skeletrophin promoter activity via an E-box-related element and was involved in downregulation of skeletrophin. An activator protein-2, which has a tumor suppressor-like role in melanoma, increased skeletrophin expression. Interestingly, exogenously expressed skeletrophin reduced melanoma cell invasion in vitro and in vivo. Colony formation in soft agar was also reduced in a RING motif-dependent manner, without affecting cell growth. We also found that skeletrophin downregulated transcription of the Met oncogene, which encodes the hepatocyte growth factor receptor and plays a role in the determination of the invasive phenotype of many malignant tumors. Finally, exogenously expressed skeletrophin, but not its RING mutant, increased transcription of Hes1 gene, a downstream effector of Notch pathway in melanoma cells. The present findings indicate that skeletrophin might be a novel suppressor factor for melanoma invasion.

Elevation of cytokeratin 19 fragment (CYFRA 21-1) in serum of patients with radiation pneumonitis: possible marker of epithelial cell damage.

Cytokeratin 19 fragment (CYFRA 21-1) level in serum have already been documented as a useful tumor marker for lung cancer. In the present study, we hypothesized that CYFRA 21-1 increases in the sera of patients with radiation pneumonitis, resulting from epithelial cell damage. We measured CYFRA 21-1 in the sera of patients with radiation pneumonitis and evaluated the correlation between CYFRA 21-1 level and severity of radiation pneumonitis as well as clinical course. We studied 16 patients diagnosed with radiation pneumonitis associated with primary lung cancer. CYFRA 21-1 levels in the sera of patients with diffuse radiation pneumonitis (n = 6) significantly increased compared to normal smokers (n = 10) or patients with local radiation pneumonitis (n = 10). CYFRA 21-1 values in sera changed according to the progression or improvement of the diffuse radiation pneumonitis. An immunohistochemical study using pulmonary tissues obtained from autopsied patients with radiation pneumonitis demonstrated that the hyaline membrane and proliferating type II pneumocytes were strongly stained by the anti-human cytokeratin 19 antibody. Our data demonstrated that CYFRA 21-1 was increased in patients with diffuse radiation pneumonitis. Since CYFRA 21-1 is widely used as a tumor marker for lung cancer, this evidence should be noted especially in irradiated patients.

Reciprocal altered expression of T-cadherin and P-cadherin in psoriasis vulgaris.

BACKGROUND: The most characteristic change in psoriasis vulgaris is markedly increased, persistent keratinocyte proliferation. The underlying mechanism of excessive epidermal growth is controversial. We previously found and reported that T-cadherin was expressed in keratinocytes and confined to the basal layer of mouse and human skin. Invasive cutaneous squamous cell carcinoma showed a loss of T-cadherin expression. Another study showed that T-cadherin was a negative growth regulator of epidermal growth factor in T-cadherin transfectant neuroblastoma cells. OBJECTIVES: To obtain insight into the role of T-cadherin in keratinocyte proliferation and to investigate further the pathogenesis of psoriasis vulgaris, we examined the expression of T-cadherin, as well as E- and P-cadherin, in psoriasis vulgaris. METHODS: Four untreated active psoriatic skin samples from psoriasis vulgaris patients and four normal human skin samples from plastic surgery were collected, cryosectioned and immunohistochemically stained by antihuman T-, P- and E-cadherin antibodies. Further, the immunofluorescence intensities of T- and P-cadherin on the basal layer of the epidermis were quantitatively measured by the histogram function of LSM 510 software installed in a Zeiss laser scanning confocal microscope. The data were statistically analysed by Student's t-test. RESULTS: It was observed that T-cadherin was weakly and discontinuously expressed on the basal layer of psoriatic skin, while it was intensively expressed on all basal keratinocytes in normal human skin. In contrast, P-cadherin was strongly expressed throughout the entire epidermal layer in psoriatic skin samples, although its expression is restricted to the basal cell layer in normal human skin. There were no obvious differences in E-cadherin expression between normal human skin and psoriatic skin. Statistical analyses showed that the immunofluorescence intensity of T-cadherin in the basal cell layer of psoriatic skin (35 +/- 9.08) was significantly decreased compared with that in normal human skin (131.75 +/- 3.49, P = 2.46 x 10(-6)). There was a significant increase (P = 0.00139) in the immunofluorescence intensity of P-cadherin in the basal layer of psoriatic skin (68.25 +/- 12.13) compared with normal human skin (26 +/- 4.90). CONCLUSIONS: The present study demonstrates that there is downregulation of T-cadherin expression and upregulation of P-cadherin expression in psoriatic skin, which are considered to be involved in the hyperproliferation of keratinocytes in psoriasis vulgaris.

KL-6, a human MUC1 mucin, is expressed early in premature lung.

KL-6, one of the MUC1 antigens, is a mucin-like high-molecular-weight glycoprotein, which is strongly expressed on type II pneumocytes. Serum levels of KL-6 have been shown to correlate with activity of interstitial pneumonia (IP). During embryonic development, MUC1 expression coincides with the onset of epithelial sheet and glandular formation. To investigate the potential role of KL-6 in lung morphogenesis, we examined KL-6 expression by immunohistochemistry on autopsied lung tissue specimens of 35 neonates and infants with gestational age from 23 to 40 weeks. Hyaline membranes (HMs) were detected in 13 of 35 cases. Simultaneously, antibody against surfactant protein A (SP-A) was employed in the study which is a distinct marker for type II pneumocytes. In all cases studied with gestational age above 23 weeks, staining for KL-6 was strongly positive in alveolar epithelial cells and in HMs found in 13 cases, whereas immunoreaction for PE10 varied depending on gestational age and duration of postnatal survival. Our findings suggest that KL-6 is expressed earlier in premature lung and may act as an important factor contributing to morphogenesis and function of developing lung in early gestation.

Expression of thyroid transcription factor-1 in 16 human lung cancer cell lines.

It has been suggested that thyroid transcription factor-1 (TTF-1) is frequently expressed in human lung cancer, especially in adenocarcinoma and small cell lung cancer, and the TTF-1 expression is closely related with the expression of surfactant protein. We hypothesized that TTF-1 is expressed in human lung cancer cell lines and its expression might be related to the expression of surfactant protein. To test this, expressions of TTF-1 and surfactant protein A (SP-A) were immunohistochemically evaluated in 16 human lung cancer cell lines. In addition, expressions of mRNAs for TTF-1 and SP-A were analyzed by reverse transcriptase-polymerase chain reaction (RT-PCR) and sequencing. As a result, nuclear staining of TTF-1 was observed in two of six adenocarcinoma cell lines, none of seven small cell lung cancer cell lines, and none of three squamous lung cancer cell lines. Among the 16 cell lines, six cell lines (PC3, LC2/Ad, A549, RERF-LC-OK, HI1017, and PC9) expressed significant amounts of mRNA for TTF-1. In contrast, cytoplasmic staining of TTF-1 was observed in five of six adenocarcinoma cell lines, in six of seven small cell lung cancer cell lines, and in all three squamous cell lung cancer cell lines. One of the two adenocarcinoma cell lines those showed positive nuclear staining and cytoplasmic SP-A staining released a significant amount of SP-A in culture supernatant. Our present study demonstrates that the frequency of TTF-1 expression in the nucleus was very low in human lung cancer cell lines; however, their cytoplasmic positivities should be further investigated.

Developmental expression of carbonic anhydrase-related proteins VIII, X, and XI in the human brain.

Three cDNA homologues of carbonic anhydrase with unknown biological functions have been reported: carbonic anhydrase-related proteins (CA-RP) VIII, X, and XI. In the present study, we produced monoclonal antibodies to these CA-RPs and studied their regional and cellular distributions in the human adult and fetal brains by immunohistochemical analysis. In the adult brain, CA-RP VIII was expressed in the neural cell body spreading to most parts of the brain. CA-RP X was expressed in the myelin sheath and its expression was shown in the cytoplasm of cultured tumor cells by immunocytochemical analysis. CA-RP XI was expressed in the neural cell body, neurites, and astrocytes in relatively limited regions of the brain. In the fetal brain, CA-RP VIII and XI were expressed in the neuroprogenitor cells in the subventricular zone as early as the 84th day of gestation and subsequently detected in the neural cells migrating to the cortex. CA-RP X first appeared in the neural cells in the cortex at the 141st day. In the choroid plexus, the epithelial cells gave CA-RP VIII and XI expressions in both adult and fetal brains. From the findings in the present study on the distribution and the developmental expression of CA-RP VIII, X, and XI in the human brain we suggest that these CA-RPs play roles in various biological process of the CNS.

[Study of the difference between the clinical diagnostic criteria and results of immunohistochemical staining].

If more than 2 lesions of cancer are observed in the lung, differences in the histology or in situ component is the basic criterion for multicentricity. In addition, remote lung mass with same histology in the absence of both distant metastasis and mediastinal lymphadenopathy is also regarded as multicentricity. We have studied the difference between the clinical diagnostic criteria and the results of immunohistochemical staining. Thirteen patients who were diagnosed as double lung cancers under the clinical of Martini et al or Cortese et al were reviewed. Of them, clinically 6 patients had synchronous double lung cancers and 7 patients had metachronous double lung cancers. Four patients in each group with combination of adenocarcinoma (AD) and bronchiolo-alveolar cell carcinoma (BAC) were studied by immunohistochemical staining. As the result, 3 patients in the former group were defined as the synchronous double lung cancers, however in the latter group, only 1 patients was defined as the metachronous double lung cancers. As for from the histological findings, if either of multiple lung cancer lesion were Noguchi's A or B typed BAC, the patients are prone to have double lung cancers. Subsequently if the histology of the both lesions were the same as AD-AD or Noguchi's C typed BAC-BAC, then the patients are prone to have the metastatic lung cancers.

The point mutation in the promoter region and the single nucleotide polymorphism in exon 1 of the cytokeratin 19 gene in human lung cancer cell lines.

The CYFRA 21-1 assay which detects the cytokeratin 19 (CK19) fragment is widely used as a tumor marker for lung cancer. We previously suggested that the failure of PCR amplification of exon 1 is closely related to the inability of the expression of mRNA for CK19, and hypothesized that point mutations might exist within exon 1. In order to prove this, sequence analysis of the promoter region and exon 1 was performed in 14 human lung cancer cell lines. Among the 14 lung cancer cell lines evaluated, point mutations within the promoter region (at -99, G-->C) of the CK19 gene were demonstrated in two cell lines (Lu135 and HI1017). In addition, point mutations within exon 1 (at 90, T-->C, Ala-->Ala and at 179, G-->C, Gly-->Ala) were also demonstrated in three cell lines (LU135, HI1017, and LC2/AD). Point mutations within the promoter region of CK19 (at -99) and within exon 1 (at 179) were confirmed by analysis of digestion by specific restriction enzymes. Since the same point mutation within exon 1 (at 179) was observed in genomes of normal volunteers, this mutation was considered as a single nucleotide polymorphism. In contrast, there were no mutations within the promoter region of exon 1 in genomes of normal volunteers. After a computer search, it was demonstrated that several transcription factors bind to the sense primer sequence which was designed for amplification of exon 1. In addition, after point mutations within the promoter region occurred (at -99), new sequences appeared to which known transcription factors (AP2) bind. In conclusion, analysis of genomic DNA for CK19 suggested that expression of mRNA for CK19 was regulated by several transcription factors which bound to the specific sequence with the promoter region of the CK19 gene. It was also suggested that the mutation in the promoter region of the CK19 gene down-regulated the expression of mRNA for CK19.

Recent progress in T-cadherin (CDH13, H-cadherin) research.

T-cadherin is a unique cadherin cell adhesion molecule that is anchored to the cell surface membrane through a glycosyl phosphatidyl inositol (GPI) moiety. The cytoplasmic domain, which T-cadherin lacks, is believed to be critical for homophilic binding through interaction with submembrane cytoskeletal proteins. Does this mean that T-cadherin is an unimportant molecule? However, the T-cadherin amino acid motif has been well conserved through evolution in vertebrates, suggesting that T-cadherin may have biological significance in higher animals. Consistent with this hypothesis, recent studies have thrown light on the relevance of T-cadherin in the fields of oncology, neurology, respirology and cardiovascular physiology. In this manuscript, we review current advances in T-cadherin research.

Immunohistochemical findings of nitric oxide synthase expression in urothelial transitional cell carcinoma including dysplasia.

Several in vitro studies have shown that nitric oxide (NO) produced by NO synthase (NOS), such as inducible NOS (iNOS) play an important role in tumor biology. We immunohistochemically examined the expression of iNOS and p53 proteins in patients with transitional cell carcinoma (TCC) of the urinary tract, including adjacent dysplastic lesions to determine the significance of the tumor behavior. Of total 94 tumors, in the present study, 41 (43.6%) tumors exhibited homogeneous immunostaining (diffuse strong positivity in tumor cells, >60%) and 53 (56.4%) tumors heterogeneous staining (variable positivity in tumor cells, 20-60%). No TCCs exhibited negative iNOS immunostaining was found. Thirty (31.9%) of 94 TCCs were positive with anti-p53 antibody, including 23 of homogeneous and 7 of heterogeneous staining. Of 23 TCCs with homogeneous p53 immunostaining, 11 tumors exhibited homogeneous iNOS immunoreaction. In the present study, dysplastic lesions adjacent to carcinomas were detected in 64 cases including 36 TCCs with homogeneous iNOS expression. All dysplastic lesions adjacent to the 36 TCCs with homogeneous iNOS immunostaining exhibited homogeneous iNOS immunostaining. No significant association between iNOS immunoreactivity and any clinicopathological factors as well as p53 immuno-reactivity were found. These in vivo findings provide evidence for frequent iNOS protein expression in TCC. In addition, our observations indicate that overexpression of iNOS expression may be one of the early events in the carcinogenesis of TCC.

Focal enhancement of expression of c-Met/hepatocyte growth factor receptor in the myocardium in human myocardial infarction.

To determine the distribution and expression level of hepatocyte growth factor (HGF) specific receptor, c-Met, in human myocardial infarction. Autopsies of 13 patients who died without heart diseases (control) and 13 patients with a history of myocardial infarction (2 h to 10 years before death). The harvested myocardial tissues were stained with hematoxylin-eosin (H&E) and immunohistochemically stained for c-Met expression by the avidin-biotin-horseradish peroxidase complex method using an antibody to c-Met. C-Met expression was only slightly increased in control subjects and in noninfarcted myocardium of the test group. In contrast, high expression was noted in the peripheral region of the myocardial infarction and in some hypertrophic myocardial cells. C-Met was not expressed in the infarcted myocardium, but overexpression was noted in the surrounding myocardial cells of blood vessels and in the subendocardium and subepicardium in a band-like pattern. The expression level of c-Met was most enhanced at the time of appearance of coagulative necrosis and least in the myocardium of subjects with old infarcts. Our results indicate that HGF preferentially reaches the ischemic regions of the myocardium and has local and direct effects on the myocardium in patients with myocardial infarction.

Primary intrapelvic seminoma in Klinefelter's syndrome.

Seminoma arising in patients with Klinefelter's syndrome is extremely rare; to our knowledge, only three cases have been reported in the English language literature. We report a case of intrapelvic seminoma in a 39-year-old man with Klinefelter's syndrome. Gross examination revealed that the tumor was a solid and irregular mass measuring 90 mm in diameter. The cut surfaces of this ill-defined tumor were yellow-white with necrotic foci. Histologically, the tumor cells were separated into lobules by branching, fibrous septa containing lymphocytes. In some parts of the tumor, a cord-like arrangement of tumor cells was present. Immunohistochemically, the tumor cells were strongly and diffusely positive for antiplacental alkaline phosphatase antibody along their cytoplasmic membranes, but negative for both chorionic gonadotrophin and alpha-fetoprotein. Based on these findings, we diagnosed this tumor as a seminoma. The testes when examined were found to be atrophic bilaterally, but with no tumor lesions. Chromosomal analysis yielded a 47XXY karyotype, compatible with Klinefelter's syndrome. These findings indicate a case of primary intrapelvic seminoma in Klinefelter's syndrome. The patient underwent intensive radiation therapy postoperatively, and he demonstrated no evidence of recurrence or metastasis during the 13-month period following surgery.

Expression of Bax and apoptosis-related proteins in human esophageal squamous cell carcinoma including dysplasia.

The rate of tumor growth depends on the balance between proliferation and death of tumor cells. It is known that Bax, caspase-3, and p53 proteins are death-promoting factors, whereas Bcl-2 protein is a death antagonist. We immunohistochemically examined the expression of Bax and apoptosis-related proteins such as caspase-3, p53, and Bcl-2 in 76 patients with human esophageal squamous cell carcinoma (SCC) including dysplasia to determine the relationship of expression of each protein to tumor behavior and patients' prognosis. No significant relationships in immunopositivity were found among these proteins in SCCs. Cytoplasmic Bax expression was exhibited in 63 cases of SCCs (82.9%). The apoptotic index of caspase-3-positive lesions was significantly higher than that of caspase-3-negative lesions in both dysplasia and SCC (P =.016, P =.012). On the other hand, the apoptotic index (1.18%) was significantly correlated with Bax overexpression in dysplasia (P =.006), but not in SCC lesions (P =.129). The patients with Bax-positive SCCs were found to have a poor prognosis by the Kaplan-Meier method (P =.043). These findings suggested that Bax expressed in dysplasia may play a role as an apoptotic factor, but that it may be functionally inactive in some cancerous lesions and thus not contribute to suppression of the tumor progression in some cases of human esophageal SCCs.

[Minocycline-induced pneumonitis presenting as multiple ring-shaped opacities on chest CT, pathologically diagnosed bronchiolitis obliterans organizing pneumonia (BOOP)].

A 39-year old woman was admitted to our hospital because of cough and abnormal shadows on chest radiographs. She had been treated for 5 months for acne vulgaris with minocycline hydrochloride (MINO). Chest computed tomographic (CT) scans showed multiple ring-shaped opacities in both lungs. Bronchoalveolar lavage disclosed an increase in the total number of cells and a marked increase of lymphocytes. A lung specimen obtained by transbronchial lung biopsy (TBLB) was pathologically diagnosed as bronchiolitis obliterans organizing pneumonia (BOOP). Withdrawal of minocycline led to rapid remission without treatment. The clinical course and histological findings for TBLB suggested that this case was minocycline-induced BOOP. Several cases with minocycline-induced pneumonitis have been reported. However, there are few reported cases of minocycline-induced BOOP, the present case being only the second found in the literature.