LY6K is a novel molecular target in bladder cancer on basis of integrate genome-wide profiling.

BACKGROUND: The aim of this study is to find a novel molecular target based on chromosomal alteration and array-based gene expression analyses in bladder cancer (BC). We investigated a cancer testis antigen, LY6K, which is located on chromosome 8q24.3. METHODS: Five BC cell lines were subjected to high-resolution array-comparative genomic hybridisation with 244 000 probes. The expression levels of LY6K mRNA were evaluated in BC cell lines and clinical BC specimens by real-time reverse transcription-PCR. The cell lines were subjected to fluorescence in situ hybridisation of LY6K. Cell viability was evaluated by cell growth, wound healing, and matrigel invasion assays. RESULTS: Typical gained loci (P<0.0001) at 6p21.33-p21.32, 8q24.3, 9q34.13, 11q13.1-q14.1, 12q13.12-q13.13, 16p13.3, and 20q11.21-q13.33 were observed in all of the cell lines. We focused on 8q24.3 locus where LY6K gene harbours, and it was the top upregulated one in the gene profile from the BC cell line. LY6K mRNA expression was significantly higher in 91 BCs than in 37 normal bladder epitheliums (P<0.0001). Fluorescence in situ hybridisation validated that the high LY6K mRNA expression was due to gene amplification in the region where the gene harbours. Cell viability assays demonstrated that significant inhibitions of cell growth, migration, and invasion occured in LY6K knock down BC cell lines; converse phenomena were observed in a stable LY6K transfectant; and LY6K knockdown of the transfectant retrieved the original phenotype from the LY6K transfectant. CONCLUSION: Upregulation of the oncogenic LY6K gene located on the gained locus at 8q24.3 may contribute BC development.

Acylated delphinidin glycosides in the blue-violet flowers of Consolida armeniaca.

Four new acylated delphinidin 3,7-glycosides were isolated from the blue-violet flowers of Consolida armeniaca as major anthocyanin pigments. The first pigment was based on delphinidin 3,7-diglucoside and was identified as delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7-O- [6-O-(4-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)-p-hydroxybenzoy l)- beta-D-glucopyranoside] by spectral methods. The other three acylated anthocyanins were based on delphinidin 3-glucoside-7-sophoroside. The second pigment was determined to be delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7-O-[2-O-(beta-D- glucopyranosyl)-6-O-(4-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)- p- hydroxybenzoyl)-beta-D-glucopyranoside]. The third pigment was elucidated to be delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7-O-[2-O-(6-O-(p- hydroxybenzoyl)-beta-D-glucopyranosyl)-6-O-(4-O-(6-O-(p-hydroxybenzoyl)- beta-D- glucopyranosyl)-p-hydroxybenzoyl)-beta-D-glucopyranoside], and the fourth as delphinidin 3-O-[6-O-(malonyl)-beta-D-glucopyranoside]-7- O-glucopyranosyl[(2-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)-6-O - (4-O-(6-O-(p-hydroxybenzoyl)-beta-D-glucopyranosyl)-p-hydroxybenzoyl)-be ta-D- glucopyranoside].

Anthocyanins from the scarlet flowers of Anemone coronaria.

Three acylated anthocyanins were isolated from the scarlet flowers of Anemone coronaria 'St. Brigid Red' along with a known pigment, pelargonidin 3-lathyroside. The structures of the acylated pigments were based on a pelargonidin 3-lathyroside skeleton acylated at different positions with malonic acid. The first pigment was identified as pelargonidin 3-O-[2-(beta-D-xylopyranosyl)-6-O-(malonyl)-beta-D-galactopyranoside], the second was pelargonidin 3-O-[2-O-(beta-D-xylopyranosyl)-6-O-(methyl-malonyl)-beta-D-galactopyranoside], and the third was (6''-O-(pelargonidin 3-O-[2''-O-(beta-D-xylopyranosyl)-beta-D-galactopyranosyl]))((4-O-(beta-D-glucopyranosyl)-trans-caffeoyl)-O-tartatryl)malonate.

(Delphinidin 3-gentiobiosyl) (apigenin 7-glucosyl) malonate from the flowers of Eichhornia crassipes.

A novel acylated delphinidin glycoside was isolated from the blue-purple flowers of Eichhornia crassipes as a major pigment, and was identified as (6'''-O-(delphinidin 3-O-(6''-O-(beta-D-glucopyranosyl)-beta-D-glucopyranosyl))) (6''-O-(apigenin 7-O-(beta-D-glucopyranosyl)))malonate by spectral methods. The three dimensional structure of this pigment was suggested from the observation of the negative Cotton effect at lambda max 535 nm that delphinidin (chromophore) and apigenin (co-pigment) occupy a folding conformation as a binary complex.

An acylated cyanidin glycoside from the red-purple flowers of Dendrobium.

A new acylated anthocyanin was isolated from the red-purple flowers of Dendrobium 'Pramot' (phalaenopsis type cv) as a major anthocyanin. The structure of this pigment was determined to be cyanidin-3-O-malonyl)-beta-D-glucopyranoside) 7,3'-di-O-(6-O-(4-O-(beta-D- glucopyranosyl)oxybenzoyl)-beta-D-glucopyranoside) mainly based on the spectroscopic evidence.

New aspects of anthocyanin complexation. Intramolecular copigmentation as a means for colour loss?

Two series of structurally related anthocyanins, extracted from the blue flowers of Evolvulus pilosus cv. Blue Daze and from the blue-purple flowers of Eichhornia crassipes, exhibit remarkable colour stabilities in aqueous solution at mildly acidic pH values. All the pigments possess the same chromophore (delphinidin), but a different pattern of glycosylation and acylation. Moreover, one of the pigments has an apigenin 7-glucoside molecule (a flavone) attached to the glycosidic chain by two ester bonds with malonic acid, instead of an aromatic acid and is the only known anthocyanin with such a structure. All the molecules studied, except one which has only a 3-gentiobioside (a disaccharide) as substituent, denote an effect of reduction in the hydration constant when compared with the parent delphinidin 3-glucoside or 3,5-diglucoside molecules, which supports the existence of intramolecular hydrophobic interactions between the chromophoric skeleton and the acyl or flavonoid groups. The role played by the sugar units on the hydrophobicity/hydrophilicity of the pigments is also discussed.

Influence of trans-cis isomerisation of coumaric acid substituents on colour variance and stabilisation in anthocyanins.

The recently isolated pigments from Petunia integrifolia and Triteleia bridgesii present a distinct feature that sheds new light on the understanding of intramolecular copigmentation of anthocyanins. These are among the infrequent anthocyanins that naturally present a coumaric acid substituent in both cis and trans forms. As a consequence, the two isomers demonstrate substantial variations of their thermodynamic and kinetic constants and also colour properties. A possible explanation for these characteristics is presented, making use of molecular modelling and taking into account the three-dimensional structures of the pigments.

Eight new anthocyanins, ternatins C1-C5 and D3 and preternatins A3 and C4 from young clitoria ternatea flowers

Eight new anthocyanins 1-8 (ternatins C1, C2, C3, C4, C5, and D3 and preternatins A3 and C4) were isolated from Clitoria ternatea flowers. By the application of chemical, UV-vis, and FABMS methods, the structures of 1-6 were postulated as delphinidin 3-malonylglucoside having 3'-GCGC-5'-G, 3'-GCGCG-5'-G, 3'-GC-5'-G, 3'-GCG-5'-G, 3'-G-5'-G, and 3'-GC-5'-GC, and compounds 7 and 8 as delphinidin 3-glucoside having 3'-GCG-5'-GCG and 3'-GCG-5'-G as side chains, respectively, in which Dp is delphinidin, G is D-glucose, and C is p-coumaric acid. The structures of the compounds 1, 3-5, and 7 were established completely by additional NMR methods.

Inhibition of cholesterol absorption by (-)N-[alpha-phenyl-beta-(p-tolyl)ethyl] linoleamide in rats.

The effect of (-)N-[alpha-phenyl-beta-(p-tolyl)ethyl] linoleamide (PTLA) on intestinal absorption of cholesterol was studied in rats. Oral administration of 15 mg PTLA to rats resulted in a significant (P less than 0.05) decrease in the radioactivity in serum and liver 4 hr after administration of labeled cholesterol. The effect of PTLA was greater on the absorption of cholesteryl oleate as compared with free cholesterol. The rate of hydrolysis of cholesteryl oleate in mucosal homogenates of rat intestine was decreased with PTLA, suggesting that the inhibition of cholesterol absorption by PTLA is related to its effect on cholesteryl ester hydrolysis in the intestine.

Investigation of the pharmacokinetics of cyclosporine in patients following long-term administration after a kidney transplant: comparison of Neoral and Sandimmune.

A crossover administration of Neoral and Sandimmune was performed in 43 renal allograft recipients who had been on cyclosporine maintenance therapy for 2 to 19 years posttransplant to investigate the pharmacokinetics of cyclosporine. Although there was no difference in C0 values (trough values) when Neoral and Sandimmune were administered at the same doses, AUC(0-4) and AUC(0-12) values of Neoral were 1.57- and 1.36-fold greater than those of Sandimmune, respectively. For both Neoral and Sandimmune, there was a high correlation between the C2 value and AUC(0-4). The Pearson's product-moment coefficient for the correlation between the C2 value and AUC(0-4) was R=0.91642. On the other hand, the correlation with the C0 value (trough value) was low (R=0.53181). During the period of the study, there was no acute rejection episode, onset of adverse drug reaction symptoms, or marked change in laboratory test values.

[Renovascular hypertension].

Renovascular disease is one of the most common causes of secondary hypertension. Recent technical advances have changed the management principles, which include a more aggressive approach to the diagnosis and treatment of renovascular hypertension (RVH). We experienced a total of 95 cases with RVH between 1958 and 1999. The mean age of all patients was 31.8 years old, ranging from 3 to 64 years. The three major basal diseases that caused RVH were fibromuscular dysplasia (34/95), arteriosclerosis (26/95), and aortitis (12/95). Ninety-two kidneys were treated in 79 of the 95 patients. The major therapeutic modalities performed were reconstruction of renal artery (6/79), nephrectomy (21/79), autotransplantation (26/79), and percutaneous transluminal angioplasty (PTA) (25/79). PTA is now the treatment of choice for the initial management of patients with RVH. Surgical treatment is generally reserved for patients in whom PTA fails. Pharmacotherapy is used on patients awaiting angioplasty or revascularization, those who are too ill for intervention, and those who have failed to respond to intervention.

[A case of subclinical IgA nephropathy and cyclosporin associated arteriolopathy diagnosed by non-episode biopsy of graft kidney after renal transplantation].

We report a case of subclinical immunoglobulin A (IgA) nephropathy and cyclosporin associated arteriolopathy following renal transplantation. A 39-year-old male with chronic glomerulonephritis received kidney transplantation from a two- human leukocyte antigen (HLA) mismatched cadaveric donor. The initial immunosuppressive therapy was triple-drug therapy with cyclosporin, prednisolone and mizoribine. Four months after transplantation, he had an acute rejection episode, and the renal function was recovered by steroid pulse and 15-deoxyspergualin therapy. Eight years after transplantation, we conducted a non-episode biopsy of the renal allograft to examine subclinical lesions. The histopathological findings showed cyclosporin associate arteriolopathy (CAA) and IgA nephropathy. There was no sign of acute or chronic rejection. At the present time, the renal function of the allograft is good. In conclusion, the non-episode biopsy of renal allograft is useful for examination of subclinical lesions.

[Histopathological findings and clinical effects for acute rejections].

We reviewed 115 cases of acute rejection following renal transplantation. All cases were diagnosed after graft biopsy, and showed histopathological evidence of acute rejection. They were treated with administration of OKT3, 15-deoxyspergualin (DSG), anti-lymphocyte globulin (ALG) or methylprednisolone (MP). All rejections were histopathologically classified according to the Banff working classification. The clinical effects of each drug were evaluated both at 1 month and 1 year following the therapy for rejection, by measurement of serum creatinine level. The effective rate both at 1 month and 1 year was related with the Banff working classification ( p < 0.0001). At 1 month after treatment, there were no significant differences between the OKT3, DSG or ALG group and MP group in cases of borderline change and AR grade I. In cases of grade II and grade III, a significant difference was observed between the OKT3 or ALG group and MP group (p < 0.05). The DSG group showed a slightly better outcome than the MP group, although the difference was not significant. In conclusion, the Banff schema is shown to be valid for classification of acute renal allograft rejection, and it is necessary to determine the treatment for acute rejection according to histopathological classification.