Ca2+-nitric oxide-cGMP signaling in rabbit parotid acinar cells.

Guanosine 3',5'-cyclic monophosphate (cGMP) is a second messenger generated in response to hormones or neurotransmitters in various tissues and cells. In parotid acinar cells, the activation of muscarinic cholinergic and beta-adrenergic receptors induces an increase in intracellular cGMP. However, the mechanism of cGMP production in parotid acinar cells has not been well elucidated.

[Percutaneous ethanol injection for unresectable pancreatic cancer-report of two cases].

Two cases of unresectable pancreatic cancer underwent percutaneous ethanol injection (PEI). The first was a 65-year-old man with a 43 x 39 mm mass in the head of the pancreas. He received PEI (total 38.

Inactivation of tumor suppressor p53 by mot-2, a hsp70 family member.

The mortalin genes, mot-1 and mot-2, are hsp70 family members that were originally cloned from normal and immortal murine cells, respectively. Their proteins differ by only two amino acid residues but exhibit different subcellular localizations, arise from two distinct genes, and have contrasting biological activities. We report here that the two proteins also differ in their interactions with the tumor suppressor protein p53.

Malignant transformation of NIH3T3 cells by overexpression of mot-2 protein.

The murine mortalin genes, mot-1 and mot-2, are members of the hsp70 family of proteins and differ from each other by only two amino acid residues. Mot-1 is expressed in normal cells and has pancytosolic cellular distribution whereas mot-2 is found in the perinuclear region of immortal cells. We report here that a high level of expression of mot-2 protein resulted in malignant transformation of cells as analysed by anchorage independent growth and nude mice assays.

Roles of the Ser146, Tyr159, and Lys163 residues in the catalytic action of 7alpha-hydroxysteroid dehydrogenase from Escherichia coli.

The Escherichia coli 7alpha-hydroxysteroid dehydrogenase (7alpha-HSDH; EC 1.1.1.

Alleviation of apoptosis by serum in Chinese hamster ovary cells ectopically expressing human Fas antigen.

Fas-mediated apoptosis is an important regulatory mechanism for the development of T-cells and prevention of oncogenesis. Here, we establish Chinese hamster ovary (CHO) cell lines which stably express Fas antigen, and analyzed apoptosis induced by anti-Fas IgM. While Fas-transfected hamster cells did not undergo apoptosis when stimulated with anti-Fas antibody in the presence of medium containing 10% serum, in reduced serum concentrations, anti-Fas antibody caused these cells to round up and detach from the culture dish.

Efficacy of ivermectin against Strongyloides stercoralis infection in jirds (Meriones unguiculatus).

The activity and distribution of ivermectin (IVM), a broad spectrum anthelmintic for various nematodes and arthropods, was tested against Strongyloides stercoralis infection in the jird (Meriones unguiculatus.) The pattern of IVM concentration in the serum of jirds after either low- (200 microg/kg) or high-dose (1000 microg/kg) treatment by intraperitoneal injection showed a typical single-peak profile with the maximum drug levels detected at 1 h followed by a rapid decline to undetectable values by 48 h posttreatment. With equivalent IVM doses, the serum levels in male jirds were significantly higher than those observed in female jirds at 1 and 12 h posttreatment (P < 0.

The regulation of cyclin D1 expression in senescent human fibroblasts.

To clarify the molecular mechanisms of cyclin D1 expression during in vitro cellular aging, we investigated the binding of nuclear protein factors to the cyclin D1 gene promoter domain in young and senescent normal human fibroblasts. The cyclin D1 promoter binding activities of nuclear protein factors from young and senescent cells were examined by the gel mobility shift assay. Our findings revealed that (i) the binding of a specific nuclear factor to the enhancer element was very weak in senescent cells; (ii) the binding of a specific nuclear factor to the CRE, which is independent of cell growth, was unchanged between young and senescent cells; (iii) nuclear factors from senescent cells did not bind to the presumptive silencer element; (iv) the binding of specific factors to the Inr (transcription initiation region) and E2F increased with growth stimulation in young cells and was weakly detectable in senescent cells; and (v) the binding of Sp1 to its promoter element occurred only in senescent cells.

Synthesis and anti-tubulin activity of ustiloxin D derivatives.

Ustiloxin D, produced by the rice plant pathogen Ustilaginoidea virens, exhibits potent anti-tubulin activity. In order to elucidate the effects of functional groups in ustiloxin D on its activity, several derivatives were synthesized and their anti-tubulin activities were estimated. The N,N-dimethylamino derivative and the 14-O-methyl derivative were inactive (IC50 > 50 microM).

Isolation and structure of an antimitotic cyclic peptide, ustiloxin F: chemical interrelation with a homologous peptide, ustiloxin B.

Ustiloxin F, a microtubule inhibitor, was isolated as a minor metabolite of Ustilaginoidea virens. The structure was determined from the spectral data and by chemical interrelation to ustiloxin B through reductive removal of the sulfoxide-containing side chain of ustiloxin B to give ustiloxin F. Ustiloxin F inhibited microtubule assembly with an IC50 value of 10.

A direct competitive ELISA for the simple, sensitive and accurate determination of diethylcarbamazine concentration in serum.

Improved methods for measurement of the anti-filariasis drug diethylcarbamazine in serum would assist in the design of effective therapy. The method evaluated in the present paper is a direct competitive ELISA which is sensitive, specific and accurate. Horseradish peroxidase-labelled diethylcarbamazine conjugate was incubated with diethylcarbamazine and anti-diethylcarbamazine antiserum over a bound second antibody.

Enhanced expression of mitochondrial genes in senescent endothelial cells and fibroblasts.

It has been suggested that some mitochondrial genes are important in cellular senescence. In order to identify the mitochondrial genes that are involved in cellular senescence, we have constructed a cDNA library from senescent human vascular endothelial cells and isolated 86 senescence-specific cDNA clones by differential screening. Among the clones, we identified four distinct mitochondrial genes including NADH dehydrogenase subunit 2 (ND2), ND3, ATPase 6 and 16S ribosomal RNA.

Triplet repeat-containing ribosomal protein L14 gene in immortalized human endothelial cell line (t-HUE4).

A cDNA encoding human 60S ribosomal subunit protein L14 (hRL14) was isolated from a human immortal endothelial cell line, t-HUE4. This cell line was established via a series of cell lines cultured in a serum-free and a protein-free medium, and a directional cDNA library has been constructed and screened in search for the genes modulating protein synthesis machinery in cell proliferation. A putative full-length clone with an open reading frame of 220 amino acids; predicted molecular weight of 23.

A novel alternatively spliced form of murine vascular endothelial growth factor, VEGF 115.

Murine immortal fibroblasts express a form of vascular endothelial growth factor (VEGF) that was cloned, characterized and named VEGF 115. It differs from VEGF 120 by 37 amino acids at the carboxyl terminus. VEGF 115-specific sequence reacted to a single transcript in mouse tissues.

Contraction and intracellular calcium-ion elevation of cultured human aortic smooth muscle cells by endothelin-1, vasoactive intestinal contractor (VIC) and the derivatives.

Effects of endothelin (ET) family peptides and their derivatives on cellular contraction and calcium-ion level were examined by using cultured human vascular smooth muscle cells (VSM). Contraction of cultured human VSM, isolated from human fetal aortic segments, was induced within 1 min after the treatment with ET-1 (100 nM) as seen in the changes of cytosolic calcium-ion localization. In parallel with the cell contraction, cytosolic calcium-ion level in the human VSM increased very rapidly and then dropped with some oscillation as determined by Anchorage Cell Analyzing System.

mdm2 and bax, downstream mediators of the p53 response, are degraded by the ubiquitin-proteasome pathway.

Upon activation in response to cellular stress or DNA damage, the p53 tumor suppressor induces the expression of gene products involved in cell cycle arrest and apoptosis. Using the proteasome-specific inhibitors, MG132 (N-acetyl-L-leucinyl-L-leucinal-L-leucinal) and lactacystin, here we show that the p53-response proteins, bax and mdm2 as well as p21, are degraded by the ubiquitin-proteasome pathway in HeLa cells. MG132 also increased expression of the three proteins in cells that lack p53, showing that stabilization of the p53 response proteins is not due to increased levels of p53 itself.

Hyperoxia induces the differentiated neuronal phenotype of PC12 cells by producing reactive oxygen species.

Neurite extension of PC12 cells induced by nerve growth factor (NGF) is a well-known model of neuronal differentiation. In this study, the incubation of PC12 cells in a 50% O2 atmosphere (hyperoxia) caused neurite extension. In these cells, amounts of differentiation-marker proteins, tyrosine hydroxylase and neurofilament M increased.

Elevated levels of mortalin expression in human brain tumors.

We have performed immunohistochemical studies of mortalin in normal and tumor human brain sections. In normal brain sections, the expression was seen mainly as being confined to neurons. Normal astrocytes showed undetectable expression of this unique member of the heat shock 70 protein family.

Nitric oxide synthase activities in mammalian parotid and submandibular salivary glands.

Nitric oxide is important as a physiological messenger molecule in various organs and cells. It is synthesized from the amino acid L-arginine by nitric oxide synthase. Here, the specific activities of nitric oxide synthase in the cytosolic fractions of rabbit, bovine, mice, rat, and guinea-pig parotid and submandibular glands were compared.

Decrease in amplified telomeric sequences and induction of senescence markers by introduction of human chromosome 7 or its segments in SUSM-1.

Introduction of human chromosome 7 by microcell-mediated chromosome transfer suppresses indefinite division of SUSM-1, an in vitro established human fibroblast line. This cell line has unusually long telomeric sequences although it lacks detectable telomerase activity. Thus, we examined whether such telomeric sequences change upon introduction of chromosome 7 or its segments.

The rescuing effect of nerve growth factor is the result of up-regulation of bcl-2 in hyperoxia-induced apoptosis of a subclone of pheochromocytoma cells, PC12h.

The rat pheochromocytoma cell line PC12 is useful for studying neuronal cell differentiation since this cell line differentiates into neuron-like cells in response to nerve growth factor (NGF). We demonstrated that PC12h cells, a subclone of PC12 cells, died under hyperoxia (50% O2). This cell death did not occur in the presence of antioxidant reagents.

Transcriptional squelching by ectopic expression of E2F-1 and p53 is alleviated by proteasome inhibitors MG-132 and lactacystin.

The transcription factors p53 and E2F-1 play important roles in the control of cell cycle progression. In transient transfection experiments, expression of E2F-1, other E2F family members, or p53 squelched transcription from cotransfected plasmids in a dose-dependent manner. Although the proteasome inhibitors MG-132 and lactacystin markedly increased the level of expression of E2F-1 and p53, these inhibitors completely alleviated squelching by both proteins.

[A case of polyneuropathy with B-CLL and HTLV-I associated myelopathy (HAM)].

We report a case of polyneuropathy associated with chronic B lymphoid leukemia (B-CLL) and HAM. A 65-year-old man had an initial onset with polyneuropathy, then myelopathy appeared. High HTLV-I antibody and anti-ganglioside antibodies were observed.