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Serum cystatin C concentrations are reported to increase in the hyperthyroid state. Serum concentrations of cystatin C and transforming growth factor-β1 (TGF-β1) were measured in patients with thyroid dysfunction, and the effects of 3,5,3′-tri-iodothyronine (T3) and TGF-β1 on cystatin C production in human hepatoblastoma (Hep G2) cells were studied. Serum concentrations of cystatin C and TGF-β1 were significantly higher in patients with Graves' disease compared with control subjects. Significantly positive correlations were observed between thyroid hormones and cystatin C, thyroid hormones and TGF-β1, and TGF-β1 and cystatin C in patients with thyroid dysfunction. Serum concentrations of cystatin C and TGF-β1 decreased after treatment for hyperthyroidism. Cystatin C mRNA levels and cystatin C secretion were increased by T3and TGF-β1 in cultured Hep G2 cells. These results suggest that serum cystatin C concentrations increase in patients with hyperthyroidism. The mechanisms for this may involve elevation of serum TGF-β1 levels and the stimulatory effects of T3and TGF-β1 on cystatin C production.

To clarify the influence of an intubation maneuver with or without premedication for an intracranial hemorrhage in an unconsciousness patient, we retrospectively analyzed 70 patients who had received intubation for unconsciousness and in whom a nontraumatic intracranial hemorrhage was found by CT over a 6-year period. They were divided into 2 groups, consisting of a drug group (n = 15), wherein drugs were used before intubation, and control group (n = 55), wherein no drugs were used before were intubation. The physical findings on admission, CT findings, Glasgow Outcome Score (GOS) at 3 months from admission were analyzed between the groups. There were no significant differences in the backgrounds of the subjects between the groups. The GOS in the control group was significantly higher than in the drug group (P < .001). In cases of intubation for unconscious patients who may have intracranial hemorrhaging, premedication is considered associated with a more favorable outcome.

Abstract: Objective: The present study examined the inhibitory effect of β₂-adrenoceptor activation on the mitogen-activated protein kinase (MAPK) cascades and the contribution of these pathways to the suppression of tumor necrosis factor (TNF)-α in lipopolysaccharide (LPS)-stimulated rat renal mesangial cells.¶Materials and Methods: Experiments were performed using cultured mesangial cells in the presence of LPS (1 μg/ml) and/or the β₂-adrenoceptor agonist, terbutaline (10^-6- 10^-8M). The levels of extracellular signal-regulated kinase-1 and 2(Erk1/2), p38, c-Jun N-terminal protein kinase (JNK) and TNF-α were estimated.¶ Results: LPS activated Erk-1/2 and p38 levels, by 4.7-fold and 1.8-fold, respectively (P &lt;0.05), which were suppressed by terbutaline (10^-6-10^-8M) in a dose dependent way. These inhibitory actions of terbutaline were prevented by the β₂-adrenoceptor antagonist, ICI 118,551(10^-6M) but not by an inhibitor of the cAMP-PKA pathway, H-89 (5 × 10^-6 M). The selective MAPK/Erk-1 inhibitor, PD98059 (10^-5M) and the specific p38 inhibitor SB203580 (10^-5M) significantly decreased LPS-induced TNF-α production in the cells.¶Conclusions: Inhibition of MAPK cascades (Erk1/2 and p38) plays an important role in the suppression of TNF-α following β₂-adrenoceptor activation but the inhibitory effect on MAPK is independent of the cAMP-PKA pathway in the mesangial cell.

Background. β₂-Adrenoceptor activation regulates tumour necrosis factor (TNF)-α and interleukin-6 (IL-6) production in cultured renal cells. However, it remains uncertain whether, in vivo, the administration of β₂-adrenoceptor agonists regulate renal TNF-α and IL-6 mRNA following lipopolysaccharide (LPS) stimulation to cause endotoxaemia. This study was performed in order to evaluate the effect of β₂-adrenoceptor agonist on renal TNF-α and IL-6 production.
Methods. Four-week-old Wistar rats pre-treated with the β₂-adrenoceptor agonist terbutaline or formoterol, and/or the β- and β₂-adrenoceptor antagonists (propanolol, ICI118,551), were injected with LPS (1 mg i.p.), and then 2, 4 or 6 h later, kidneys (cortex, medulla), spleen, thymus and plasma were collected to assay TNF-α and IL-6 mRNA levels and their respective protein release.
Results. Administration of β₂-adrenoceptor agonists suppressed TNF-α mRNA expression in the whole kidney, by 61% (P&lt;0.05), as well as plasma, spleen and thymus TNF-α protein and mRNA expression 2 hours after injection of LPS. On the other hand, although IL-6 levels in plasma, spleen and thymus mRNA expression were suppressed significantly by administration of β₂-adrenoceptor agonists, the basal- and LPS-induced IL-6 mRNA levels in the whole kidney were increased 1.6- and 1.2-fold (P&lt;0.05), respectively, by treatment with β₂-adrenoceptor agonists. β₂-Adrenoceptor agonist suppressed LPS-induced TNF-α mRNA expression by 35% (P&lt;0.05) and stimulated LPS-induced IL-6 mRNA expression by 1.5-fold (P&lt;0.05) in the medullary region of kidney.
Conclusions. β₂-Adrenoceptor agonists down-regulate renal TNF-α mRNA expression following LPS-induced endotoxaemia. This effect was particularly apparent in the renal medulla. IL-6 mRNA expression in the renal medulla was up-regulated by the agonists whereas plasma, spleen and thymus IL-6 levels were completely inhibited by the agonist, which suggests the existence of tissue specific regulation of IL-6 production in the kidney by β₂-adrenoceptor activation.

beta(2)-Adrenoceptor activation regulates tumour necrosis factor (TNF)-alpha and interleukin-6 (IL-6) production in cultured renal cells. However, it remains uncertain whether, in vivo, the administration of beta(2)-adrenoceptor agonists regulate renal TNF-alpha and IL-6 mRNA following lipopolysaccharide (LPS) stimulation to cause endotoxaemia. This study was performed in order to evaluate the effect of beta(2)-adrenoceptor agonist on renal TNF-alpha and IL-6 production.

A full-length complementary DNA (cDNA) clone (pTK-3) encoding an isoform of Mg2+-dependent protein phosphatase β (MPPβ-4) was isolated for the first time from a mouse melanocyte cDNA library. It was strongly suggested that the mRNA corresponding to the pTK-3 insert was a splicing variant of a single pre-mRNA that also encodes MPPβ-1 and -2 (T. Terasawa, T. Kobayashi, T. Murakami, M. Ohnishi, S. Kato, O. Tanaka, H. Kondo, H. Yamamoto, T. Takeuchi, and S. Tamura, 1993, Arch. Biochem. Biophys. 307, 342-349). The amino acid sequence of MPPβ-4 differed from those of MPPβ-1 and -2 only at the carboxyl terminal region. Analysis by reverse transcriptase polymerase chain reaction (RT-PCR) revealed that MPPβ-4 mRNA was expressed only in testis and intestine and not in other mouse tissues tested. Specific expression of the mRNA signals of two other isoforms of MPPβ, MPPβ-3 and -5 (a novel isoform), in testis and intestine was also demonstrated by the RT-PCR. The carboxyl terminal region of MPPβ-5 was found to have a chimera structure composed of part of MPPβ-1 and part of MPPβ-3. The recombinant MPPβ-3 and -4 and the putative MPPβ-5 expressed in Escherichia coli cells exhibited Mg2+-dependent and okadaic acid-insensitive protein phosphatase activities. It was demonstrated that the mRNA expression levels of MPPβ-3, -4, and -5 alter according to the maturation of mouse testis. These resuits suggest that the complex structure of MPPβ isoforms and their tissue- and developmental stage-specific expression reflect the variety of their physiological functions.Copyright 1995, 1999 Academic Press, Inc.

Mouse P19 embryonal carcinoma cells in aggregation culture in the presence of 10(-6) M retinoic acid followed by monolayer culture differentiate into nerve and glial cells. In this study, we demonstrated that the neurofilament-L (NF-L) mRNA and protein levels of these cells were enhanced in accordance with their retinoic acid-induced neural differentiation. Okadaic acid (OA) treatment of the cells markedly suppressed this differentiation-dependent NF-L gene expression increase and neurite outgrowth of the cells. Similar results were obtained when tautomycin was used instead of OA, suggesting that inhibition of protein phosphatase(s) is involved in the suppression of neural differentiation. OA treatment did not affect the NF-L gene transcription level, determined by the nuclear run-on transcription assay, but it did reduce the stability of both the 3.5- and 2.3-kilobase NF-L mRNAs. The expression and activity levels of protein phosphatase 2A (PP2A) and 2B (PP2B) but not protein phosphatase 1 (PP1) in P19 cells increased in accordance with the enhanced NF-L gene expression. The presence of OA in the culture medium during the course of the neural differentiation caused a reduced PP2A activity but not PP1 and PP2B activities of the cell extracts. On the other hand, both PP1 and PP2B activities but not PP2A activity of cell extracts were suppressed by the addition of cyclosporin A or FK506 in the culture medium. However, both cyclosporin A and FK506 treatments affected neither NF-L gene expression nor neurite outgrowth. These results demonstrate that the OA treatment inhibits the differentiation-dependent increase in NF-L gene expression by destabilizing its mRNAs and suggest that PP2A plays key roles in the differentiation-dependent enhanced expression of the NF-L gene and is the point of the action of OA.

Human cytosolic aldehyde dehydrogenase 1 (ALDH1) plays a role in the biosynthesis of retinoic acid that is a modulator for gene expression and cell differentiation. Northern blot analysis showed that liver tissue, pancreas tissue, hepatoma cells, and genital skin fibroblast cells expressed high levels of ALDH1. Sequence analysis showed that the 5'-flanking region contains a number of putative regulatory elements, such as NF-IL6, HNF-5, GATA binding sites, and putative response elements for interleukin-6, phenobarbital and androgen, in addition to a noncanonical TATA box (ATAAA) and a CCAAT box. Functional characterization of the 5'-regulatory region of the human ALDH1 gene was carried out by a fusion to the chloramphenicol acetyltransferase gene. A construct containing 2.6 kilobase pairs of the 5'-flanking region was efficiently expressed in hepatoma Hep3B cells, but not in erythroleukemic K562 cells or in fibroblast LTK- cells, which do not express ALDH1. Within this region, we define a minimal promoter (-91 to +53) that contains positive regulatory elements. The study using site-directed mutagenesis demonstrated that the CCAAT box region is the major cis-acting element involved in basal ALDH1 promoter activity in Hep3B cells. Gel mobility shift assays showed that NF-Y and other octamer factors bound CCAAT box and an octamer motif sequence, but not GATA site existing in the minimal promoter region. Two additional DNA binding activities associated with the minimal promoter were found in the nuclear extract from Hep3B cells, but not from K562 cells. These results offer the possible molecular mechanism of the cell type-specific expression of ALDH1 gene.

In our study, we found a recognizable pattern of the midface in three retinoblastoma patients with interstitial deletion of chromosome 13 in lymphocytes. Further photographs of patients reported by other authors confirmed our conclusion that the midface in children with interstitial deletion of 13q from q12 to q22, always involving q14, is characterized by prominent eyebrows, broad nasal bridge, bulbous tip of the nose, a large mouth with a thin upper lip, and a long philtrum. This midface pattern could not be recognized in patients with normal karyotypes, suggesting that the critical segment for the midface phenotype as well as for retinoblastoma may exist in band 13q14.