Transport of magnesium by two isoforms of the Na+-Ca2+ exchanger expressed in CCL39 fibroblasts

Cytoplasmic concentrations of Ca2+ ([Ca2+]i) and Mg2+ ([Mg2+]i) were measured with fluorescent indicators in CCL39 cells, a cell line established from Chinese hamster lung fibroblasts, transfected with complementary deoxyribonucleic acid (cDNA) of the Na+-Ca2+ exchanger isolated either from canine heart (NCX1) or from rat brain (NCX3). Raising extracellular [Mg2+] to 10 mM increased Mg2+ influx and the resultant change in [Mg2+]i (delta[Mg2+]i) was monitored with furaptra under Ca2+-free conditions. In control (vector-transfected) cells, delta[Mg2+]i at 45 min was similar with or without extracellular Na+ (130 mM or 0 mM) and when [Na+]i was raised by 1 mM ouabain treatment. delta[Mg2+]i in NCX1-transfected cells was attenuated significantly in the presence of 130 mM Na+, but became comparable to (or slightly larger than) that in control cells on either removal of extracellular Na+ or treatment with 1 mM ouabain. Cells expressing NCX3 showed an intermediate dependence of delta[Mg2+]i on Na+, probably reflecting a lower degree of expression of the exchanger protein. Extracellular Na+-dependent changes in [Ca2+]i (measured with fura-2 in the presence of extracellular Ca2+ and 10 microM ionomycin, a Ca2+ ionophore) were minimal in control cells, marked in the NCX1-transfected cells and intermediate in the NCX3-transfected cells. These results suggest that the Na+-Ca2+ exchanger (either NCX1 or NCX3) can transport Mg2+ and may play a role in the extrusion of magnesium from cells.     

Morphometric comparison of human nerve cells: pyramidal motor system

We compared morphological and morphometric data on various motor neurons in the human pyramidal system using the modified Klüver-Barrera staining method with extremely minimized shrinkage ratio and an image-analyzer. We classified motor neurons in the human pyramidal system into three groups according to the measurement data. This report may be of interest to better understand the process of nerve conduction in the human pyramidal system.     

Comparison of protective immunity elicited by recombinant vaccinia viruses that synthesize E or NS1 of Japanese encephalitis virus.

Immunization with recombinant vaccinia viruses that specified the synthesis of Japanese encephalitis virus (JEV) glycoproteins protected mice from a lethal intraperitoneal challenge with JEV. Recombinants which coexpressed the genes for the structural glycoproteins, prM and E, elicited high levels of neutralizing (NEUT) and hemagglutination inhibiting (HAI) antibodies in mice and protected mice from a lethal challenge by JEV. Recombinants expressing only the gene for the nonstructural glycoprotein, NS1, induced antibodies to NS1 but provided low levels of protection from a similar challenge dose of JEV. Antibodies to the NS3 protein in postchallenge sera, representing the degree of infection with challenge virus, were inversely correlated to NEUT and HAI titers and levels of protection. These results indicate that although vaccinia recombinants expressing NS1 can provide some protection from lethal JEV infection, recombinants expressing prM and E elicited higher levels of protective immunity.     

Pathology of colorectal adenomas: a colonoscopic survey.

The size, histological type, and grade of dysplasia of a large series of colorectal adenomas removed by colonoscopic polypectomy were matched against other variables such as anatomical site, age, sex, and number of adenomas per patients. Special emphasis was placed on the criteria for grading dysplasia in adenomas and the possible significance of severe dysplasia as a selective marker for increased colorectal cancer risk. The results showed that small adenomas (mostly with mild dysplasia) were evenly distributed throughout the colorectum but that adenomas showing severe dysplasia (mostly the larger tumours, greater than 10 mm diameter) were concentrated in the left colon and rectum, particularly the sigmoid part which is also the segment with the highest risk of colorectal carcinoma in high risk populations. Severe dysplasia in adenomas appears to be a selective histopathological marker for increased colorectal cancer risk. It is closely linked with increasing age and numbers of adenomas per patient, with the large adenomas and particularly those with a villous component in their histology. Severe dysplasia and multiple adenomas could be valuable markers for selecting from the total adenoma population those most deserving of close surveillance in follow-up cancer prevention programmes. Conceptually it would appear advantageous to think in term of the dysplasia-carcinoma sequence in the colorectum rather than the polyp-cancer or adenoma-carcinoma sequence. The implications of these results in the study of the aetiology of colorectal cancer are discussed.     

Combination therapy of cyclosporine with steroid inhibits γ-interferon and interleukin-1 gene expression at the level of mRNA synthesisin vivo

The present study examined the effect of cyclosporine (CsA) administered with steroidin vivo on the capacity of peripheral blood mononuclear cells (PBMC) from kidney transplant recipients to generate cytokines and their gene expression at the level of messenger RNA (mRNA). PBMC from CsA-prednisolone (Pred)-treated recipients displayed 66.9% inhibition (54.3±12.4 IU/ml;N=42;P<0.01) of -interferon (-IFN) production compared with normal individuals (134.6±18.6 IU/ml;N=23). Azathioprine (Az)-Pred-treated recipients displayed significantly less inhibition of -IFN generation (96.0±16.1 IU/ml;N=22;P<0.05) than CsA-treated patients. Macrophages (m) from CsA-Pred-treated recipients displayed 60.0% inhibition (5.1±0.7 U/ml;N=20;P<0.01) of interleukin-1 (IL-1) production compared with normal individuals (13.0±2.9 U/ml;N=21). These results were confirmed by the experiments using cDNA probe for -IFN or IL-1 (, ). High levels of -IFN mRNA in phytohemagglutinin (PHA)-stimulated PBMC or IL-1() mRNA in lipopolysaccharide (LPS)-stimulated m were present in normal individuals but not in CsA-treated recipients as judged by hybridization to a cloned human -IFN or IL-1() cDNA probe. These studies demonstrated that combination therapy of CsA with steroid inhibits both -IFN and IL-1 gene expression at the level of mRNA at physiological concentrations.     

Identification of cembratriene-4,6-diol as antitumor-promoting agent from cigarette smoke condensate

Cigarette smoke condensate (CSC) was separated into severalfractions and each was tested for an inhibitory effect on theearly antigen (EA) of Epstein-Barr virus (EBV) which can beinduced by 12-0-tetradecanoylphorbol-13-acetate (TPA) in Rajicells. Two diastereoisomers of 2,7,11-cembratriene-4,6-diol(- and ß-CBT) were isolated from the neutral fractionsof CSC and these showed potent inhibitory effects on the inductionof EBV-EA by TPA. The doses of - and ß-CBT requiredfor 50% inhibition of EBV-EA induction by TPA were 7.7 and 6.7µg/ml, respectively, which are comparable with those ofretinoic acid, a potent inhibitor of induction of epideral ornithinedecarboxylase (ODC) activity and tumor promotion by TPA in mice.Application of - and ß-CBT to mouse skin prior totreatment with TPA inhibited TPA-induced ODC activity. The degreeof inhibition was dependent on the dose and application of 16.5µmol/mouse of - and ß-CBT resulted in a 50 and40% reduction, respectively, of the maximum of the ODC activityinduced as a result of treatment with TPA. In initiation-promotionexperiments, -CBT markedly inhibited the promoting effect ofTPA on skin tumor formation in mice which were initiated with7,12-dimethylbenz[a]anthracene, but ß-CBT was foundto be less effective. Application of 3.3 µmol of -CBT40 min prior to treatment with TPA (1 µg) resulted ina 53% reduction in the number of papillomas per mouse. Our presentdata suggest that EBV-EA inhibition assay using Raji cells iseffective for the first screening of inhibitors of tumor promotion,and provide evidence that CSC contains antitumor-promoting agentsin addition to carcinogenic and tumor-promoting agents alreadyreported.     

Effect of vitamin E on the induction and evolution of enzyme-altered foci in the liver of rats treated with diethylnitrosamine

The effects of dietary vitamin E (VE) on the steps of hepatocarcinogenesis,the induction and growth of -glutamyltranspeptidase (GGT)-positivefoci and their evolution into persistent nodules, were analyzedin the liver of rats treated with diethylnitrosamine (DEN).The induction of GGT-positive foci was inhibited by a diet containing0.36–1.5% VE given after initiation with 200 mg/kg bodyweight (b.w.) DEN for 6 weeks with partial hepatectomy (PH)on week 3. The numbers and areas of GGT-positive foci were enhancedby diets containing 036 and 0.72% VE, given for 1 week afterinitiation with 10 mg/kg b.w. DEN and PH, followed by selectionby 0.02/ 2-acetylaminofluorene (AAF) and carbon tetrachloride(CCl⁴), but these were not enhanced by a diet containing 1.5%VE. Remodeling of hyperplastic nodules was not affected by thediet containing 0.72% VE given after initiation with DEN andselection for 12 weeks. The staining characteristics of GGTwere different between remodeling and persistent nodules, exceptfor those of the glutathione-S-transferase placental form (GST-P).The results obtained suggest that VE could prevent the veryearly events during hepatocarcinogenesis, the induction of phenotypicallyaltered foci, but could no longer affect the later stages, theevolution of foci into persistent nodules.     

Persistent effect of a low dose of preadministered diethylnitrosamine on the induction of enzyme-altered foci in rat liver

The effect of a low dose of preadministered diethylnitrosamine(DEN) on the induction of enzyme-altered foci in the liversof male full-grown Fischer 344 rats was studied. As a pretreatment,DEN at a dose of 10 mg/kg body wt was injected i.p. At varioustimes after DEN pretreatment a complete initiation, consistingof administration of the same dose of DEN by the same routein rats subjected to partial hepatectomy (PH), was performed,followed by application of selection pressure. Enzyme-alteredfoci stained with -glutamyltrans-peptidase (-GTP) and glutathioneS-transferase placental form (GST-P) were then assayed. Decreasesin the numbers and areas of foci in the rats which receivedsaline + PH 14 or 28 days after DEN pretreatment were observedin comparison with rats which received saline + PH immediatelyafter DEN. On the other hand, the numbers and areas of fociwere not decreased in rats which received the complete initiation,consisting of DEN + PH, at various times after DEN pretreatmentwhen compared with rats which received these at the same timeas the DEN pretreatment. This persistent effect of DEN pretreatmenton the complete initiation lasted up to 182 days after the timeof DEN pretreatment. In this experiment, GST-P was found tobe a more sensitive marker for the detection of putative preneoplasticliver-cell foci than -GTP.     

Neural map of interaural phase difference in the owl''s brainstem.

Neurons of the barn owl's (Tyto alba) nucleus laminaris, the first site of binaural convergence, respond in a phase-locked fashion to a tone delivered to either ear. It may take longer to elicit phase-locked spikes from one ear than from the other. This disparity in delay differs from neuron to neuron and is independent of tonal frequency. In binaural stimulation, neurons respond best when sound in one ear leads that in the other by an amount equal to their delay disparities but opposite in sign. This condition causes simultaneous arrival of phase-locked spikes from the two sides. Laminaris neurons can thus be described as coincidence detectors. The phase of a tone-induced evoked potential, termed "neurophonic," varies systematically with position in nucleus laminaris. From dorsal to ventral within the nucleus, the phase delay of a contralaterally elicited potential decreases and that of its ipsilateral counterpart increases. Therefore, if the neurophonic delay is due to the delay of phase-locked spikes, an orderly representation of delay disparities is shown. Because they act as coincidence detectors, laminaris neurons should show selectivity for interaural phase difference based on their place in the nucleus. Thus, nucleus laminaris presumably measures and maps interaural phase differences by using the principles of delay lines and coincidence detection.