High spatial-density, cladding-pumped 6-mode 7-core fiber amplifier for C-band operation.

We present a C-band 6-mode 7-core fiber amplifier in an all-fiberized cladding-pumped configuration for space division multiplexed transmission supporting a record 42 spatial channels. With optimized fiber components (e.g. passively cooled pump laser diode, pump coupler, pump stripper), high power multimode pump light is coupled to the active fiber without any noticeable thermal degradation and an average gain of 18 dB and noise figure of 5.4 dB are obtained with an average differential modal gain of 3.4 dB.

Histone variant macroH2A1.2 is mono-ubiquitinated at its histone domain.

Histone macroH2A1.2 (macroH2A) is an unusual histone H2A variant with a large non-histone macrodomain at its carboxyl terminal. MacroH2A1.2 is enriched in facultative heterochromatin, including inactivated X chromosomes in mammalian females and senescence-associated heterochromatin foci. We show here that a small population of macroH2A1.2 is mono-ubiquitinated in human HeLa cells. Mass spectrometry analysis revealed that the specific targeting sites for the mono-ubiquitination are Lys115 and Lys116 of the histone domain. A corresponding Lys119 conserved in histone H2A is also mono-ubiquitinated by Ring protein in the polycomb group complex. We suggest that the mono-ubiquitination of macroH2A1.2 and histone H2A has similar or synergistic implications, but that the multiple ubiquitination sites in macroH2A1.2 might confer a variety of functions upon macroH2A1.2 to modulate chromatin states.

[Intractable pneumothorax for pleuritis carcinomatosis effectively treated with bronchial embolization: report of a case].

A 57-year-old female was admitted with compliant of cough and body weight loss. Chest X-ray and thoracic computed tomography (CT) scan revealed a collapsed lung and pleural effusion. We diagnosed a pleulitis carcinomatosis. After right chest tube drainage was performed, she developed right intractable pneumothorax. It was occluded endobronchially by the placement of vascular embolization coils and histoacryl. This method is thought to be an effective treatment for intractable pneumothorax patients in endstage of lung cancer.

Clinical significance of vascular endothelial growth factor expression in gastric cancer.

Vascular endothelial growth factor (VEGF) is an angiogenic factor in human cancer tissue. To clarify the clinical significance of this factor, we investigated the VEGF expression in early and advanced gastric cancer. This study included analysis of data on 243 patients with gastric cancer, including 118 in the early stage and 125 in the advanced stage. VEGF was immunohistochemically stained. Of 243 tumors, 102 (42%) were VEGF-positive. The VEGF-positive gastric cancers were larger, more invasive, and classified in the more advanced stage than VEGF negative ones. Patients with VEGF-positive cancers had significantly lower survival rates than did those with negative ones, both in early and advanced stages (P < 0.05, P < 0.01, respectively). The VEGF-positive isolates had more hematogenous metastases than VEGF-negative ones. Multivariate analysis revealed VEGF to be an independent prognostic factor and independent risk factor for liver metastasis. The VEGF expression in cancer cells can serve as a pertinent prognostic indicator both in early and advanced gastric cancer.

Nuclear expression of the Y-box binding protein, YB-1, as a novel marker of disease progression in non-small cell lung cancer.

Transcription factor Y-box binding protein 1 (YB-1) that binds to the inverted CCAAT box is involved not only in transcription of various genes but also in cell proliferation and DNA repair. We determined whether localization of YB-1 in either the nucleus or cytoplasm could serve as a prognostic marker for patients with non-small cell lung cancer (NSCLC). In 196 NSCLC patients, expression of YB-1 protein in the nucleus or cytoplasm was immunohistochemically evaluated. Of the 196 tumors examined, 88 (44.9%) were positive for YB-1 expression in the nucleus. Nuclear YB-1 expression significantly correlated with T factor, lymph node metastasis, and stage of the disease. Patients with a nuclear YB-1 tumor had a poorer prognosis than did those with a cytoplasmic YB-1 tumor in all of the NSCLC patients (P = 0.0494) and in patients with squamous cell carcinoma (P = 0.0313) but not in patients with adenocarcinomas. Nuclear localization of the YB-1 protein may prove to be an important factor of disease progression for patients with NSCLC, in particular, in cases of squamous cell carcinoma.

[Angiogenesis and macrophage infiltration in Borrmann type IV gastric cancer].

The clinical significance of angiogenesis was investigated in Borrmann type IV gastric cancer. Tumors with high microvessel density (MVD) often metastasized to the liver and lymph nodes. A significant correlation was recognized between macrophage infiltration and MVD. However, MVD was not a prognostic factor. Peritoneal dissemination was a prognostic factor in Borrmann type IV gastric cancer. Thus, angiogenesis plays an important role in the metastasis, but not prognosis in Borrmann type IV gastric cancer.

A novel transformation suppressor, Pdcd4, inhibits AP-1 transactivation but not NF-kappaB or ODC transactivation.

Pdcd4 is a novel transformation suppressor that is highly expressed in promotion-resistant (P-) mouse epidermal JB6 cells but not in susceptible (P+) cells. Overexpression of pdcd4 cDNA in stably transfected P+ cells rendered cells resistant to tumor promoter-induced transformation, indicating that elevated expression of Pdcd4 protein is sufficient to suppress neoplastic transformation. To determine whether Pdcd4 suppresses neoplastic transformation through inhibiting known transformation required events, we examined the possibility that pdcd4 inhibited the activation of AP-1 or NF-kappaB dependent transcription or of ornithine decarboxylase (ODC) activity. Activation of AP-1-dependent transcriptional activity was inhibited by pdcd4 expression in a concentration dependent manner. In contrast, Pdcd4 slightly increased NF-kappaB-dependent transcription and did not alter ODC enzymatic activity. Previous studies suggested that activation of AP-1 was required for P+ cell transformation as well as for tumor promotion in vivo. These results indicate that Pdcd4 functions as a transformation suppressor, possibly through inhibiting AP-1 activation in combination with other factors such as enhancing NF-kappaB activation. Pdcd4 may thus constitute a useful molecular target for cancer prevention.

FASCIATA genes for chromatin assembly factor-1 in arabidopsis maintain the cellular organization of apical meristems.

Postembryonic development of plants depends on the activity of apical meristems established during embryogenesis. The shoot apical meristem (SAM) and the root apical meristem (RAM) have similar but distinct cellular organization. Arabidopsis FASCIATA1 (FAS1) and FAS2 genes maintain the cellular and functional organization of both SAM and RAM, and FAS gene products are subunits of the Arabidopsis counterpart of chromatin assembly factor-1 (CAF-1). fas mutants are defective in maintenance of the expression states of WUSCHEL (WUS) in SAM and SCARECROW (SCR) in RAM. We suggest that CAF-1 plays a critical role in the organization of SAM and RAM during postembryonic development by facilitating stable maintenance of gene expression states.

Gastric cancer with high telomerase activity shows rapid development and invasiveness.

Telomerase activity was reported to be activated in most immortal cells and cancers. As the clinical significance of telomerase activity in human gastric cancer is controversial, we investigated this activity using a telomeric repeat amplification protocol. The telomerase activity was tentatively defined by strength of activity as follows: 3+, observed with 0.06 microg of protein; 2+, observed with 0.6 microg of protein; 1+, observed with 6 microg of protein; 0, not observed under these three conditions. Telomerase activity was detected in 35 of 39 (89.7%) gastric cancer specimens. Tumors with high telomerase activities (2+/3+) tended to have a deeper invasion, lymphatic and vascular invasion, lymph node metastasis, liver metastasis, and peritoneal dissemination, as compared to findings in case of low telomerase activities (-/1+). Thus, telomerase activity of gastric cancer tissue may reflect the malignant potential of the tumor and intensive postoperative care might be required for such patients.

PCNA connects DNA replication to epigenetic inheritance in yeast.

Formation of a heterochromatin-like structure results in transcriptional silencing at the HM mating-type loci and telomeres in Saccharomyces cerevisiae. Once formed, such epigenetically determined structures are inherited for many mitotic divisions. Here we show that mutations in the proliferating cell nuclear antigen (PCNA), an essential component at the DNA replication fork, reduced repression of genes near a telomere and at the silent mating-typelocus, HMR. The pol30-8 mutant displayed coexistence of both repressed (pink) and de-repressed (white) cells within a single colony when assayed with the ADE2 gene inserted at HMR. Unlike pol30-8, the pol30-6 and pol30-79 mutants partially reduced gene silencing at telomeres and the HMR and synergistically decreased silencing in cells lacking chromatin assembly factor 1 (CAF-1). All silencing defective mutants showed reduced binding to CAF-1 in vitro and altered chromatin association of the CAF-1 large subunit in vivo. Thus, PCNA participates in inheritance of both DNA and epigenetic chromatin structures during the S phase of the cell cycle, the latter by at least two mechanisms.

The N-terminal domains of histones H3 and H4 are not necessary for chromatin assembly factor-1- mediated nucleosome assembly onto replicated DNA in vitro.

An in vitro reconstitution system for the analysis of replication-coupled nucleosome assembly is described. In this "two-step system," nucleosome assembly is performed in a separate reaction from DNA replication, wherein purified newly replicated DNA remains noncovalently marked for subsequent chromatin assembly factor-1 (CAF-1)-dependent nucleosome assembly. Because the nucleosome assembly is performed separately from the DNA replication step, this system is more versatile and biochemically tractable when compared with nucleosome assembly during simian virus 40 (SV40) DNA replication. The N-terminal domains of histones H3 and H4 play an important but redundant function in nucleosome assembly in the budding yeast, Saccharomyces cerevisiae. It had been proposed that at least one tail of histone H3 or H4 is required for replication-coupled nucleosome assembly. However, we demonstrate that the N-terminal domains of both histone H3 and H4 are dispensable for CAF-1-mediated formation of nucleosome cores onto newly replicated DNA in vitro. CAF-1 and each of its individual subunits stably bound to recombinant (H3.H4)(2) tetramers lacking the N-terminal domains of both H3 and H4. Therefore, the N-terminal tails of histone H3 and H4 that contain the specific acetylation sites are not necessary for CAF-1-dependent nucleosome assembly onto replicated DNA. We suggest that the histone acetylation may be required for a CAF-1 independent pathway or function after deposition, by marking of newly replicated chromatin.

Overexpression of the heat shock protein HSP70 family and p53 protein and prognosis for patients with gastric cancer.

The cell synthesis of heat shock proteins is increased by a variety of environmental and pathophysiological stressful conditions. The 70-kD heat shock protein family (HSP70 family) which constitutively expresses hsc70 and heat-inducible hsp70 is thought to be involved in protein-protein interactions, including oncogene products. We investigated the HSP70 family expression and biological behavior of gastric cancer, and its relation to p53 overexpression. Expressions of HSP70 and p53 in 164 primary gastric tumors were determined immunohistochemically. Exploratory data were analyzed on a set of 164 primary gastric cancers, and we constructed in prognostic significance of the HSP70 expression level and the relation to p53 overexpression. Expression of HSP70 (hsc70 and hsp70) were detected in nuclei and/or cytoplasm of cancer cells. Western blotting analysis showed that hsc70 and hsp70 were both expressed in five gastric cancer cell lines. Immunohistochemically stained positive cells of HSP70 varied from 0 (very weak) to 100%, in each case. The median level of positive cell rate was 19.0%. A HSP70 expression of over 19.0% was related to the differentiated tissue type of gastric cancer, but not to other clinicopathological factors. There was no difference in survival rates in subjects with higher and lower groups of HSP70 expression. HSP70 expression was also not related to p53 overexpression in the nuclei and p53 overexpression-related poorer prognosis. Our findings show that the expression of HSP70 is not associated with tumor advance-related characteristics or with the prognosis of gastric cancer. Measurements of HSP70 expression do not appear to be a useful prognostic marker.

Heterogeneity and clinical role of thymidine phosphorylase activity in gastric cancer.

We examined the dThdPase activity in primary gastric cancer and metastatic lymph nodes from human subjects. The dThdPase activities were significantly higher in primary tumors than in the normal gastric wall, particularly in the center of the tumor rather than in the periphery (P<0.01). Tumors with a high dThdPase activity often had venous invasion (P<0.01). The dThdPase activities were significantly higher in metastatic lymph nodes than in the nodes without metastasis (P<0.01). The intratumoral heterogeneity of dThdPase activity was identified and cancer cells with high dThdPase activity may indicate that metastasis will likely occur.

Natural course of combined limb and palatal tremor caused by cerebellar-brain stem infarction.

After infarction of the left superior cerebellar peduncle and dentate nucleus, a patient developed tremor of the left upper limb beginning on the twelfth day followed by palatal tremor appearing 10 months after infarction. Surface electromyogram revealed a difference in the frequency of the tremor in the upper limb and soft palate. When the palatal tremor appeared, brain magnetic resonance T2-weighted images revealed high signal intensity of the contralateral, right inferior olivary nucleus. Subsequently, when the amplitude of palatal tremor became less severe, the high olivary signal intensity subsided whereas the hypertrophy of the nucleus remained. This patient provides useful information on the pathogenesis of skeletal and palatal tremor with brain stem or cerebellar lesions based on the differences in the onset and frequency of tremors and morphologic changes in the inferior olive.

Replication-dependent marking of DNA by PCNA facilitates CAF-1-coupled inheritance of chromatin.

Chromatin assembly factor 1 (CAF-1) is required for inheritance of epigenetically determined chromosomal states in vivo and promotes assembly of chromatin during DNA replication in vitro. Herein, we demonstrate that after DNA replication, replicated, but not unreplicated, DNA is also competent for CAF-1-dependent chromatin assembly. The proliferating cell nuclear antigen (PCNA), a DNA polymerase clamp, is a component of the replication-dependent marking of DNA for chromatin assembly. The clamp loader, replication factor C (RFC), can reverse this mark by unloading PCNA from the replicated DNA. PCNA binds directly to p150, the largest subunit of CAF-1, and the two proteins colocalize at sites of DNA replication in cells. We suggest that PCNA and CAF-1 connect DNA replication to chromatin assembly and the inheritance of epigenetic chromosome states.

Membrane type 1-matrix metalloproteinase is involved in the formation of hepatocyte growth factor/scatter factor-induced branching tubules in madin-darby canine kidney epithelial cells.

Matrix metalloproteinases (MMPs) are believed to be involved in morphogenesis. Association of MMPs in a model of kidney tubulogenesis was studied using Madin-Darby canine kidney (MDCK) epithelial cells in an in vitro morphogenetic system. MDCK cells form branching tubules in three-dimensional collagen gel matrix in the presence of hepatocyte growth factor (HGF). The addition of specific MMP inhibitor BB-94 and tissue inhibitor MMP (TIMP)-2 but not TIMP-1 to such collagen gel cultures reduced the formation of branching tubules induced by HGF. The induction of membrane-type 1-matrix metalloproteinase (MT1-MMP) mRNA expression was observed in MDCK cells cultured in the collagen gel. Stable expression of MT1-MMP antisense RNA interfered with the tubule formation of MDCK cells induced by HGF-collagen gel culture. These observations implicate MT1-MMP in kidney tubulogenesis and TIMP-2-specific inhibition suggests a direct role of MT1-MMP rather than a gelatinase A-mediated effect.

Steroid-refractory severe ulcerative colitis responding to cyclosporine and long-term follow-up.

We report a case of steroid-refractory ulcerative colitis, treated with cyclosporine, in a 38-year-old woman with a 13-year history of ulcerative colitis. No remission was achieved with treatments that included intravenous hyperalimentation, sulfasalazine, and intensive parenteral prednisolone therapy for 4 weeks. Intravenous infusion of cyclosporine was performed because the patient refused to undergo surgery. Her condition improved dramatically and colectomy was avoided. She has been maintained on oral cyclosporine and azathioprine since steroids were discontinued, and she has remained in clinical and endoscopic remission for 2 years. The side effects were not significant, but mild paresthesia in both hands and mild hypertension, which was controlled by anti-hypertensives. Cyclosporine seems to be an effective treatment for patients with steroid-refractory severe active ulcerative colitis in whom colectomy seems inevitable. We believe further clinical trials of the treatment are warranted.

Study of the pharmacokinetics of cefpirome sulphate in the elderly.

OBJECTIVE: To determine the appropriate method of administration of the cephem antibiotic cefpirome sulphate in elderly patients. METHOD: We studied cefpirome's pharmacokinetics in patients with urinary tract infections. Patients received cefpirome sulphate 0.5 g by intravenous drip infusion over 30 mins. RESULTS: Patients with a creatinine clearance rate (Ccr) of 80 ml/min had an AUC of 96.7 microg.h/ml and a T1/2 of 2.36 h, whereas those with Ccr of 40-80 ml/min had an AUC of 172.0 microg.h/ml and a T1/2 of 3.45 h and those with Ccr of < 40 ml/min had an AUC of 152 microg.h/ ml and a T1/2 of 4.86 h. CONCLUSION: These results indicate that decreased kidney function can cause increases in the AUC and T1/2 of cefpirome. Thus in elderly patients and perhaps also in other patients with decreased kidney function, cefpirome should be administered at an initial dose of 0.5 g.

Molecular epidemiology and changing distribution of genotypes of measles virus field strains in Japan.

Based on phylogenetic and restriction fragment length polymorphism analyses of the hemagglutinin and nucleoprotein gene sequences, measles virus strains obtained in western Japan were divided into two types. Type 1 isolates have largely replaced type 2 isolates during the last 10 years in the area surveyed.

Rat TAFII31 gene is induced upon programmed cell death in differentiated PC12 cells deprived of NGF.

Typical programmed cell death (PCD) requires de novo macromolecular synthesis and shares common morphological changes referred to as apoptosis. To elucidate the molecular mechanism of apoptosis, we isolated cDNA clones that are induced in differentiated PC12 cells deprived of NGF by differential display method. Among such clones, homology searches revealed that the one clone encodes the rat TATA-binding-protein-associated factor TAFII31, a component of TFIID, and a transcriptional coactivator of the p53 protein. Northern analysis of various organs in human showed one band in heart, brain, skeletal muscle and pancreas, whose size is approximately 1.1 kb which identical to that of human TAFII31 mRNA, although the size of rat human TAFII31 mRNA is approximately 2.7 kb. The deduced amino acid sequence of the rat TAFII31 was 77% identical to that of the human TAFII31. Northern analysis of various organs in adult mice showed that expression levels of TAFII31 mRNA were strong in heart but weak in spleen, although this gene is ubiquitously expressed.