[Peripheral 5-FU blood concentration after high-dose injection into the hepatic artery: potential preventive effect on extrahepatic metastatic foci].

To clarify the effect of high-dose 5-FU injection into the hepatic artery (1,000 mg/m2 weekly) on liver metastases of colorectal cancer, the peripheral venous 5-FU concentration was measured in two groups of patients, one which had undergone hepatectomy and the other which had not. The area under the concentration-time curve (AUC) was calculated and the preventive effect of 5-FU on extrahepatic lesions was examined. The peripheral venous 5-FU concentration and AUC were higher in patients who received the drug via the hepatic artery after hepatectomy, and 5-FU was effective for the prevention of extrahepatic lesions as well as against recurrence in the residual liver.

New screening methods for chemicals with hormonal activities using interaction of nuclear hormone receptor with coactivator.

The endocrine system exerts important functions in a multitude of physiological processes including embryogenesis, differentiation, and homeostasis. Xenobiotics may modify natural endocrine function and so affect human health and wildlife. It is necessary, therefore, to understand the degree to which xenobiotics can disrupt endocrine systems. The key targets of endocrine disruptors are nuclear hormone receptors, which bind to steroid hormones and regulate their gene transcription. We have developed relevant assay systems based on the ligand-dependent interaction between nuclear hormone receptor and coactivator. The coactivators used in this study contained CBP, p300, RIP140, SRC1, TIF1, and TIF2. By two hybrid assay in yeast, the interactions of estrogen receptor with RIP140, SRC1, TIF1, and TIF2 were detected and they were completely dependent on the presence of estrogen. Specificity of this assay was assessed by determining the effect of steroids, known estrogen receptor agonists, and phytoestrogens. The pattern of response to chemicals were consistent with estrogenic activity measured by other assay systems, indicating that this assay system is reliable for measuring estrogenic activity. In addition, we carried out in vitro binding studies: GST pull-down assay and surface plasmon resonance analysis. The estrogen receptor also bound to coactivator in response to chemicals depending on their estrogenic activity in vitro. These data demonstrate that the measurement of interaction between steroid hormone receptor and coactivator serves as a useful tool for identifying chemicals that interact with steroid receptors.

Cloning and mapping of ZNF231, a novel brain-specific gene encoding neuronal double zinc finger protein whose expression is enhanced in a neurodegenerative disorder, multiple system atrophy (MSA).

A novel brain-specific gene, neuronal double zinc finger protein (ZNF231), was cloned and mapped. We used the high-density cDNA filter method to analyze the gene-expression profile in brains with multiple system atrophy (MSA). MSA is a sporadic progressive neurodegenerative disease characterized clinically by cerebellar symptoms, parkinsonism, autonomic dysfunction, or their various combinations, but its pathogenesis has yet to be clarified. In total, 8300 cDNA clones were screened, and a novel gene, ZNF231, was identified, whose expression was elevated in cerebella of patients with MSA. Its transcript is approximately 16 kb long and encodes an open reading frame of 3926 amino acid residues that has several interesting motifs; two glycine-proline dipeptide repeats (aa 22-32 and aa 61-74), a pair of homologous C8 double zinc finger motifs (aa 169-226 and aa 465-521), a leucine zipper motif (aa 561-582), a SH3 domain-binding motif (aa 825-831), two nuclear targeting signals (aa 1011-1028 and aa 1071-1091), two glutamine-rich domains (aa 2428-2473 and aa 3775-3804), and a histidine-rich domain (aa 3597-3682). These features suggest that the new gene encodes a nuclear protein or transcription regulator. Northern blot and RT-PCR analyses showed that its expression is specific to the brain and apparently restricted to the neurons. Elevation of ZNF231 expression may be involved in the pathogenesis of multiple system atrophy. The gene for ZNF231 is located on chromosome 3p21.

Synthesis of 15 alpha-hydroxyestrogen 15-N-acetylglucosaminides.

The synthesis of 15-N-acetylglucosaminides of 15 alpha-hydroxyesterone, 15 alpha-hydroxyestradiol, and 15 alpha-hydroxyestriol (estetrol) is described. The latter two were prepared by condensation of 2-acetamido-1 alpha-chloro-1,2-dideoxy-3,4,6-trio-O-acetyl-D-glucopyranose with appropriately protected 15 alpha-hydroxyestrogens by the Koenigs-Knorr reaction employing cadmium carbonate as a catalyst. Subsequent removal of protecting groups with methanolic potassium hydroxide provided the desired conjugates. 15 alpha-Hydroxyestrone 15-N-acetylglucosaminide was synthesized from the corresponding 15 alpha-hydroxyestradiol derivative by Jones oxidation followed by brief alkaline hydrolysis. These conjugates underwent enzymatic hydrolysis with beta-N-acetylglucosaminidase from Jack beans to produce 15 alpha-hydroxyestrogens.

Analysis of zinc and other elements in rat pancreas, with studies in acute pancreatitis.

Determination of the concentration of certain elements makes it possible to investigate the physiology of the pancreas. We used X-ray fluorescence to determine the concentrations of zinc and other elements in the pancreas of normal (control) rats and those with cerulein-induced pancreatitis. Ten elements (Zn, Ni, Fe, P, Ca, Cl, S, K, Ti, and Mn) were detected in controls. In the early stage of acute pancreatitis, the pancreatic concentrations of Zn, Ni, Fe, and P were significantly decreased (P < 0.05) and those of Ca and Cl were significantly increased (P < 0.05), compared with control levels. However, levels of S, K, and Ti did not differ significantly from the control values. Mn was detected in only some samples. The serum levels of Zn and Fe were significantly elevated (P < 0.05) in acute pancreatitis. These observations indicate that Zn and these other nine elements could play an important role in acute pancreatitis.

A facile one-step synthesis of delta 1,4-3-keto bile acid esters by iodoxybenzene and benzeneselenic anhydride.

A facile one-step conversion of stereoisomeric methyl 3-hydroxycholanoates to 1,4-dien-3-one, by treatment in boiling toluene with iodoxybenzene catalyzed by benzeneselenic anhydride, is described. The direct oxidation-dehydrogenation at C-3 is applicable to other cholanoates (hyodeoxycholic, chenodeoxycholic, ursodeoxycholic, deoxycholic, and cholic) when the hydroxyl groups at C-6, C-7, or C-12 are protected. Without protection at C-6 and C-7, the products from hyodeoxycholic, chenodeoxycholic, ursodeoxycholic, and cholic esters are complex mixtures, whereas, methyl deoxycholate yields the 1,4-dien-3,12-dione cleanly.

Impaired metabolism of arachidonate in selenium deficient animals.

Arachidonate-induced respiratory distress in mice was aggravated in a selenium deficient group as compared with a selenium supplemented one. The aggravation in selenium deficient mice may be due to enhanced platelet aggregation and suppressed formation of prostacyclin (PGI2) in the arterial all.

Distribution of monoamine-containing terminals and fibers in the central nervous system of the chicken.

In the mesencephalon of the chicken, the two large groups of CA terminals and fibers appeared to correspond to the dorsal and ventral bundles of the CA containing fibers in the rat. These ran rostrally in the medial forebrain bundle and appeared to terminate within the thalamus, hypothalamus, preoptic region and corpus striatum. 5-HT containing terminals and fibers also were observed in the mesencephalon and appeared to run rostrally in the medial part of the medial forebrain bundle. In comparison with the CA terminals, however, 5-HT terminals were for the greater part, difficult to observe in many areas of the brain. Based on the observation concerning postnatal ontogeny of monoamine-containing neurons, the chicken brain appears to acquire behavioral and functional maturity at an earlier stage after birth than does that of the rat. Effect of reserpine, nialnmide, L-Dopa and 5-HTP on the monoamine-containing terminals and fibers are briefly discussed.