[A case of mixed adenoneuroendocrine carcinoma of the distal bile duct].

We diagnosed distal cholangiocarcinoma in a 76-year-old woman who was then treated by subtotal stomach-preserving pancreaticoduodenectomy. Histopathological examination revealed a well-differentiated tubular adenocarcinoma on the side of the bile duct, and a neuroendocrine carcinoma in an area outside the bile duct where the tumor had infiltrated. Immunohistochemical staining identified homology between cytokeratins and MUC, indicating a similar origin. This report discusses problems associated with diagnosis and treatment by summarizing 22 patients who underwent curative resection and subsequently had a confirmed prognosis.

Rice leaf hydrophobicity and gas films are conferred by a wax synthesis gene (LGF1) and contribute to flood tolerance.

Floods impede gas (O2 and CO2 ) exchange between plants and the environment. A mechanism to enhance plant gas exchange under water comprises gas films on hydrophobic leaves, but the genetic regulation of this mechanism is unknown. We used a rice mutant (dripping wet leaf 7, drp7) which does not retain gas films on leaves, and its wild-type (Kinmaze), in gene discovery for this trait. Gene complementation was tested in transgenic lines. Functional properties of leaves as related to gas film retention and underwater photosynthesis were evaluated. Leaf Gas Film 1 (LGF1) was identified as the gene determining leaf gas films. LGF1 regulates C30 primary alcohol synthesis, which is necessary for abundant epicuticular wax platelets, leaf hydrophobicity and gas films on submerged leaves. This trait enhanced underwater photosynthesis 8.2-fold and contributes to submergence tolerance. Gene function was verified by a complementation test of LGF1 expressed in the drp7 mutant background, which restored C30 primary alcohol synthesis, wax platelet abundance, leaf hydrophobicity, gas film retention, and underwater photosynthesis. The discovery of LGF1 provides an opportunity to better understand variation amongst rice genotypes for gas film retention ability and to target various alleles in breeding for improved submergence tolerance for yield stability in flood-prone areas.

Pneumonia risks in bedridden patients receiving oral care and their screening tool: Malnutrition and urinary tract infection-induced inflammation.

AIM: Pneumonia develops in bedridden patients even when they are receiving oral care. However, the pneumonia risk in bedridden patients remains unclear, and no screening tool has been developed to assess this risk by using daily hospital data. METHODS: We retrospectively examined pneumonia risk factors by analyzing the records of 102 bedridden patients receiving oral care. RESULTS: Body mass index, peripheral blood hemoglobin, and serum concentrations of total protein, albumin, total cholesterol and uric acid in the pneumonia group (n = 51; mean age 73.4 years) were lower than those in the non-pneumonia group (n = 51; mean age 68.1 years). In the univariate analysis, body mass index; leukocytosis; high C-reactive protein; low levels of hemoglobin, total protein and albumin (<3.5 g/dL); and urine bacteria were associated with the development of pneumonia. Furthermore, in the multivariate analysis, low levels of albumin and urine bacteria were independently associated with pneumonia. We developed a bedridden patient pneumonia risk (BPPR) score using these two risk factors to assess pneumonia risk. We applied scores of zero (0) or one (1) according to the absence or presence of the two risk factors and summed the scores in each patient. The proportion of pneumonia patients increased with increasing BPPR score when the patients were divided into three groups - low, moderate and high risk - according to the BPPR score (0, 1 or 2, respectively). CONCLUSIONS: Malnutrition, urinary tract infection-induced inflammation and anemia were associated with pneumonia in bedridden patients. BPPR scoring might be useful for assessing pneumonia risk and managing affected patients. Geriatr Gerontol Int 2018; 18: 714-722.

Clinicopathological significance of ganglioside DSGb5 expression in renal cell carcinoma.

Disialosyl globopentaosylceramide (DSGb5) is a ganglioside originally isolated from tissue extracts of renal cell carcinoma (RCC) with metastasis. Previous in vitro experiments have suggested that DSGb5 promotes metastasis by enhancing the migration of RCC cells and downregulating NK cell cytotoxicity against RCC cells. In this study, we investigated the clinicopathological significance of DSGb5 expression in RCC and outcomes of RCC patients. A total of 156 RCC patients who underwent surgical treatments at our hospital from January 2007 through December 2012 were analyzed in this study. The expression of DSGb5 in RCC specimens was examined by immunohistochemical staining with monoclonal antibody 5F3. The immunostaining intensity of RCC tissues was assessed in comparison with that in benign renal tubules as an internal positive control. The relationship between DSGb5 expression and clinicopathological characteristics was investigated and recurrence free survival following surgery was evaluated. Microvascular invasion was observed in 68% (n = 19/28) and in 45% (n = 58/128) of the DSGb5 high expression group and low expression group, respectively (p = 0.031). Of 156 patients with a median follow up of 51 months, 18 patients (12%) developed metastasis following surgery. Patients in the DSGb5 high expression group showed significantly lower recurrence-free survival as compared with those in the DSGb5 low expression group (log-rank P = 0.047). In the present study, DSGb5 expression was associated with microvascular invasion in RCC tissues, and patients with DSGb5 high expression showed significantly lower recurrence-free survival rates. These findings suggest that DSGb5 expressed in RCC is correlated with metastasis and is a potential predictor for identifying patients who experience metastasis after surgery.

Significance of Expression of Complement C4d in Esophageal Squamous Cell Carcinoma.

BACKGROUND/AIM: Esophageal squamous cell carcinoma (ESCC) is one of the most difficult malignancies to cure. C4d is a degradation product of the classical complement pathway and is suggested as an early diagnostic marker for other SCCs. The purpose of this study was to clarify the association of complement C4d with ESCC. PATIENTS AND METHODS: Immunohistochemical staining for C4d was performed on surgical specimens obtained from 114 patients with ESCC. RESULTS: Positive C4d expression was observed in 70 (61.4%) cases and negative expression in 44 (38.6%) cases. There was a significant inverse correlation between C4d expression and depth of tumor invasion (p=0.0001), lymph node metastasis (p=0.011), lymphatic invasion (p=0.033), and TNM stage (p=0.0021). Kaplan-Meier analysis showed that negative C4d expression tended to lead to shorter overall survival (p=0.232). CONCLUSION: C4d expression in ESCC might be useful in developing treatment strategies or suppression of ESCC.

Synthesis and biological evaluation of novel FK228 analogues as potential isoform selective HDAC inhibitors.

Novel C4- and C7-modified FK228 analogues were efficiently synthesized in a highly convergent and unified manner. This synthesis features the amide condensation of glycine-d-cysteine-containing segments with d-valine-containing segments for the direct assembly of the corresponding seco-acids, which are key precursors of macrolactones. The HDAC inhibition assay and cell-growth inhibition analysis of the synthesized analogues revealed novel aspects of their structure-activity relationship. This study demonstrated that simple modification at the C4 and C7 side chains in FK228 is effective for improving both HDAC inhibitory activity and isoform selectivity; moreover, potent and highly isoform-selective class I HDAC1 inhibitors were identified.

[A CASE OF PRIMITIVE NEUROECTODERMAL TUMOR OF THE TESTIS].

A 44-year-old man discovered a swollen right testis more than 4 years earlier. He was brought to our hospital because of abdominal pain and vomiting. Enhanced computed tomography (CT) showed a swollen right testis, lung nodules, and swollen retroperitoneal and mediastinal lymph nodes. The swollen lymph nodes compressed the duodenum, causing ileus. HCG, HCG-ß, and AFP levels were normal, but the LDH level was high (2,933 IU/L). A diagnosis of testicular cancer with lung and lymph node metastases was made, and a right orchidectomy was performed. However, the pathological diagnosis was unclear, and it was necessary to consult another pathologist, but this took .6 weeks. While awaiting the pathological diagnosis, the patient was given chemotherapy with two 3-week courses of BEP. On pathological examination, the tumor consisted of small round cells with a rosette-like arrangement. Cartilage and keratinized tissues were also present. Immunohistochemical staining was positive for CD56, synaptophysin, vimentin, GFAP, and CD99 (MIC2), but negative for AE1/AE3, OCT-4, chromogranin, INI-1, and desmin. The patient was then diagnosed as having a primitive neuroectodermal tumor and teratoma. The metastatic lymph nodes decreased in size after chemotherapy; therefore, two further courses of BEP were added. However, CT showed disease progression. The patient refused further therapy and returned home. Eight months later, he was hospitalized because of swollen retroperitoneal and mediastinal lymph nodes and ileus. Despite treatment with radiation therapy, which resulted in decreased lymph nodes, the patient died. This was a very rare case, the first such case in Japan.

A pivotal role of Krüppel-like factor 5 in regulation of cancer stem-like cells in hepatocellular carcinoma.

In hepatocellular carcinoma (HCC), there exists a highly tumorigenic subset of cells defined by high expression of CD44 and CD133 that has been reported to contain cancer stem-like cells (CSCs). Krüppel-like factor 5 (KLF5) regulates many factors involved in cell cycle, migration, inflammation, angiogenesis and stemness and has cancer-promoting effects in some cancers. While some reports have indicated that KLF5 may have important roles in regulation of CSCs, what role, if any, KLF5 plays in regulation of CSCs in HCC remains to be elucidated. Flow cytometric analysis of CD44 and CD133 in HCC cell lines revealed subpopulations of CD44(High)/CD133(High) and CD44(Low)/CD133(Low) cells. We subsequently sorted these subpopulations and identified KLF5 as a gene that is significantly upregulated in CD44(High)/CD44(High) cells via RNA sequencing using next generation sequencing technology. Moreover, KLF5 overexpression enriched the CD44(High)/CD133(High) subpopulation and, consistent with the up-regulation of CD44(High)/CD133(High) cells, KLF5 overexpressing cells were more resistant to anti-cancer drugs and displayed enhanced colony-formation capacity. By contrast, knock-down of KLF5 by siRNA diminished the CD44(High)/CD133(High) subpopulation. When KLF5 was acetylated by TGF-ß1, the KLF5-mediated CD44(High)/CD133(High) subpopulation enrichment was abrogated. Oppositely, ectopic expression of an acetylation-deficient KLF5 mutant further increased CD44(High)/CD133(High) subpopulations as compared to cell expressing wild-type KLF5. These findings provide novel mechanistic insight into a pivotal role for KLF5 in the regulation of CSCs in HCC.

Ganglioside, disialosyl globopentaosylceramide (DSGb5), enhances the migration of renal cell carcinoma cells.

About one third of renal cell carcinoma (RCC) patients exhibit metastasis upon initial presentation. However, the molecular basis for RCC metastasis is not fully understood. A ganglioside, disialosyl globopentaosylceramide (DSGb5), was originally isolated from RCC tissue extracts, and its expression is correlated with RCC metastatic potential. DSGb5 is synthesized by GalNAc α2,6-sialyltransferase VI (ST6GalNAcVI) and is expressed on the surface of RCC cells. Importantly, DSGb5 binds to sialic acid-binding Ig-like lectin-7 (Siglec-7) expressed on natural killer (NK) cells, thereby inhibiting NK-cell cytotoxicity. However, the role of DSGb5 in RCC progression remains obscure. To address this issue, we used ACHN cells derived from malignant pleural effusion of a patient with metastatic RCC. Using the limiting dilution method, we isolated three independent clones with different DSGb5 expression levels. Comparison of these clones indicated that the cloned cells with high DSGb5 expression levels exhibited greater migration potential, compared to the clone with low DSGb5 expression levels. In contrast, DSGb5 expression levels exerted no significant effect on cell proliferation. We then established the ACHN-derived cell lines that stably expressed siRNA against ST6GalNAcVI mRNA or control siRNA. Importantly, the ST6GalNAcVI-knockdown cells expressed low levels of DSGb5. We thus demonstrated the significantly decreased migration potential of the ST6GalNAcVI-knockdown cells with low DSGb5 expression levels, compared to the control siRNA-transfected cells expressing high DSGb5 levels, but no significant difference in the cell proliferation. Thus, DSGb5 expression may ensure the migration of RCC cells. We propose that DSGb5 expressed on RCC cells may determine their metastatic capability.

Docetaxel, cisplatin and 5-fluorouracil chemotherapy with concurrent radiation for unresectable advanced urethral carcinoma.

The present case report describes two cases of unresectable advanced primary urethral carcinoma. Case 1 was a 61-year-old man with squamous cell carcinoma of the urethra that directly invaded into the perineal skin. Case 2 was a 64-year-old woman with adenocarcinoma, and solitary lung and lymph node metastases. Both patients received docetaxel/cisplatin/5-fluorouracil chemotherapy with concurrent radiation. The docetaxel/cisplatin/5-fluorouracil chemotherapy regimen consisted of docetaxel (75 mg/m(2) ) on day 2, cisplatin (100 mg/m(2) ) on day 2 and 5-fluorouracil (1000 mg/m(2) ) on days 1 to 5 and was repeated every 4 weeks. Complete response was achieved after two cycles of chemotherapy combined with radiation in case 1. In case 2, complete response was achieved locally after six cycles of chemotherapy combined with radiation, but the solitary lung lesion remained viable. We experienced two cases with advanced urethral carcinoma for whom docetaxel/cisplatin/5-fluorouracil chemotherapy chemotherapy with radiation was effective.

Involvement of intestinal intraepithelial lymphocytes in turnover of intestinal epithelial cells: morphological and functional alterations due to daily administration of FK506.

To elucidate the interaction of intestinal intraepithelial lymphocytes (IELs) with intestinal epithelial cells (IECs), we investigated alterations of IECs by activating or inactivating IELs. The stimulation of IELs with anti-mouse CD3 monoclonal antibody induced massive apoptosis of IECs. Changes in IECs and IELs from mice that received daily administration of FK506 for 14days were investigated. IELs, particularly TCR-γδ⁺ IELs, were reduced in cell number, and a decrease of cytotoxic activity was observed. Under this condition, loss of apoptotic cells at the tips of villi and delayed turnover of IECs were detected. The expressions of alkaline phosphatase and CD98 amino acid transporters on IECs were decreased. Furthermore, abnormal skeletal organization of villi and weakened binding of IECs to the basement membrane were shown. These results suggest that inactivated IECs, which should be led to apoptosis, remained. It was strongly suggested that IELs participated in IEC turnover.

Higher pAkt expression predicts a significant worse prognosis in glioblastomas.

phosphorylated-Akt (pAkt) plays an important role in tumorigenesis through promotion of cell survival by inhibiting apoptosis and mediating cell proliferation. Higher expression of pAkt has been reported to be associated with an unfavorable prognosis in several malignant tumors. In this study, the prognostic value of pAkt expression was investigated in glioblastomas by using immunohistochemical methods. Tissue sections obtained from 64 patients with glioblastoma were evaluated. The mean and median follow-up period was 16.2 +/- 12.4 and 12 months, respectively (range: from 1 to 62 months). pAkt expression levels were determined by immunohistochemical staining and evaluated for cell positivity. Positive staining was defined when more than 50% of the tumor cells were stained in each section. The correlation between expression of pAkt and overall survival rate was assessed. Glioblastomas showed either or both cytoplasmic and nuclear positive findings for pAkt. A total of 29.7% (19/64) of tissue specimens had greater than 50% positivity. The median survival periods of the patients with pAkt positive and negative tumor were 10 and 14 months, respectively. Two years overall survival rate of the pAkt positive and negative patients were 0% and 24.4%, respectively. Survival rate of the patients with pAkt positive tumor was significantly lower than that of the patients with pAkt negative tumors (p = 0.004). Multivariate analysis showed that extent of surgery was the strongest factor for survival (p = 0.01) and the pAkt expression was the secondly strongest factor (p = 0.06). These results suggest that the higher expression of pAkt the poorer prognosis in patients with glioblastoma.

The ADAXIALIZED LEAF1 gene functions in leaf and embryonic pattern formation in rice.

The adaxial-abaxial axis in leaf primordia is thought to be established first and is necessary for the expansion of the leaf lamina along the mediolateral axis. To understand axis information in leaf development, we isolated the adaxialized leaf1 (adl1) mutant in rice, which forms abaxially rolled leaves. adl1 leaves are covered with bulliform-like cells, which are normally distributed only on the adaxial surface. An adl1 double mutant with the adaxially snowy leaf mutant, which has albino cells that specifically appear in the abaxial mesophyll tissue, indicated that adl1 leaves show adaxialization in both epidermal and mesophyll tissues. The expression of HD-ZIPIII genes in adl1 mutant increased in mature leaves, but not in the young primordia or the SAM. This indicated that ADL1 may not be directly involved in determining initial leaf polarity, but rather is associated with the maintenance of axis information. ADL1 encodes a plant-specific calpain-like cysteine proteinase orthologous to maize DEFECTIVE KERNEL1. Furthermore, we identified intermediate and strong alleles of the adl1 mutant that generate shootless embryos and globular-arrested embryos with aleurone layer loss, respectively. We propose that ADL1 plays an important role in pattern formation of the leaf and embryo by promoting proper epidermal development.

Nuclear survivin expression predicts poorer prognosis in glioblastoma.

Survivin is a member of the inhibitor of apoptosis family, and is expressed in various malignant tumors. Survivin overexpression has been reported to be a poorer prognostic factor in various malignancies. However, the prognostic value of survivin expression in patients with glioblastoma is still controversial. Therefore, in this study the role of survivin as a predictor for survival was investigated in patients with glioblastoma. Tissue specimens were obtained from 66 patients with glioblastoma treated with radiotherapy. Survivin expression was detected by an immunohistochemical method. Nuclear and cytoplasm survivin scores were defined by using the cell positivity and staining intensity. The scores were defined as follows, 0 (no staining), 1 (less than 50% of cell positivity and any staining), 2 (more than 50% of cell positivity and weak to moderate intensity) and 3 (more than 50% of cell positivity and strong intensity). The correlation between survivin scores and the overall survival rate was evaluated. Nuclear and cytoplasm survivin staining were noted in 47 and 58 patients, respectively. The number of patients with nuclear survivin score of 0, 1, 2 and 3, were 19 (28.8%), 26 (39.4%), 9 (13.6%) and 12 (18.2%), respectively. The 3-year overall survival rate of the nuclear survivin score 3 was 0%, significantly lower than the 11.6% of the nuclear survivin score

Kinetic determination of cysteine on flow injection system by utilizing catalytic complexation reaction of Cu(II) with 5,10,15,20-tetrakis (4-N-trimethylammino-phenyl) porphyrin.

A kinetic method for the determination of cysteine on flow injection system is described. Cysteine was found as a catalyst for the complexation reaction of Cu(II) with an ultra sensitive colorimetric reagent of 5,10,15,20-tetrakis (4-N-trimethylammino-phenyl) porphyrin (TTMAPP) (varepsilon=4.8x10(5)cm(-1)M(-1) at 433nm), which was used as the indicator reaction in this paper. Soret band of either the porphyrin or the produced complex (varepsilon=4.6x10(5)cm(-1)M(-1) at 411nm) can be used for detection. The continuous flow injection system greatly enhanced the analytic precision and efficiency of the kinetic method, giving a relative standard deviation of 0.73% for a 0.1mugml(-1) cysteine with 10 injections at a throughput of 30h(-1). The detection limits (3S/N) in this case was 15ngml(-1) and the working dynamic range was over 25ngml(-1) to 1mugml(-1). Sugars, organic acids and amino acids that are possible in coexistence with cysteine could be tolerated at high concentrations. This method was critically compared with the Ellman's reagent in the determinations of cysteine contents of three pharmaceutical injections for hepatic diseases and one permanent wave agent and showed better applicability in the respect of matrix interferences.