PCB contamination assessment of yusho patients by using preserved human umbilical cord.

PCB concentrations in umbilical cord preserved from the time when Yusho patients and healthy subjects gave birth were examined. The total concentration of the 12 DL-PCB isomers ranged from 130 to 12,000 pg/g in the umbilical samples, was about 700 pg/g around 1950 but began to increase in the mid 1960s, reached about 12,000 pg/g between 1968 and 1970 immediately after the Yusho incident. However, the DL-PCB concentration was high between 1968 and 1970 in not only the designated Yusho patients but also healthy subjects, and the maximum DL-PCB concentration was close between the two groups.

The effect on immunocytes of anodic oxide titanium after hydrothermal treatment.

All dental root implants come in contact with the oral epithelium, and many complex factors are found to arise in this region. In order to perform a successful dental root implantation, it is necessary to clarify the interaction of the dental root implant material with the host defense mechanisms involved in the specific and nonspecific immune responses to many antigens in oral bacteria and their components. Recently, focusing on developing the dental root implant, the Nikon Corporation improved the surface characteristics of pure titanium even further by developing a hydroxyapatite (HA) layer formed on an anodic titanium oxide film containing Ca and P via hydrothermal treatment (SA treatment). However, since little is known about the effect of SA-treated pure titanium (HA/Ti) on the defense mechanisms of the oral membrane epithelium, we investigated (1) the in vitro proliferation of murine splenic B lymphocytes on the surface of HA/Ti in the presence of three lipopolysaccharide (LPS) concentrations and (2) interleukin-1alpha (IL-1alpha) production by the reaction of human peripheral blood mononuclear cells (PBM cells) on the surface of HA/Ti under the same concentrations. After culture, murine splenic lymphocytes were measured by uptake of 3H-thymidine, and cytokine release (IL-1alpha) from PBM cells was measured by ELISA. Results showed that HA/Ti had hardly any effect on the LPS-induced proliferation of B lymphocytes and IL-1alpha production. In vitro investigations of the effects of HA/Ti on the LPS-induced proliferation of murine splenic B lymphocytes and IL-1alpha from PBM cells might be a useful way of elucidating the defense mechanism between implants and the oral epithelium.

Histomorphometric evaluation of the thin hydroxyapatite layer formed through anodization followed by hydrothermal treatment.

By the previously described method of electrochemical and hydrothermal reaction, a thin hydroxyapatite (HA) layer of 1 microm thickness was formed on machined, grit-blasted, and titanium plasma-sprayed implants, the surfaces of which were equipped with a gap zone of 0.15 mm in depth. These implants, together with HA and titanium plasma-sprayed implants as control materials, were placed in dog mandibles for 4 weeks. Histomorphometrical comparison was performed to examine the effects of the thin HA layer and the surface topography on bone formation. The roughened implants, especially the grit-blasted implants, were surrounded with thin bone newly formed along the rough surfaces and showed higher bone apposition than the smooth implants. The gap zone of the HA plasma-sprayed implant was repaired with new bone that had vertically extended from the surrounding bone. The thin HA layer had as much osteoconduction as a plasma-sprayed HA coating but showed significantly different bone response. The results suggest that bone formation on an HA film is affected by degradation in living tissue that is related to the crystallinity and the chemical composition of the HA film itself.

Mechanical and histological investigation of hydrothermally treated and untreated anodic titanium oxide films containing Ca and P.

In a previous study a new method for forming thin hydroxyapatite (HA) layers on titanium was described. Titanium was anodized at 350 V in an electrolyte solution containing sodium beta-glycerophosphate and calcium acetate, and an anodic titanium oxide film containing Ca and P (AOFCP) was formed on the surface. Then numerous HA crystals were precipitated on the AOFCP during hydrothermal treatment in high-pressure steam at 300 degrees C. In this study three types of hydrothermally treated films differing in amounts of precipitated HA crystals and tensile adhesive strength, and untreated films were histologically and mechanically investigated in a transcortical rabbit femoral model for 8 weeks of implantation using light microscopy, scanning electron microscopy (SEM), and push-out tests. Machined titanium and HA ceramics served as control materials. The push-out shear strength and bone apposition of the AOFCP significantly increased after hydrothermal treatment, and were equivalent to those of HA ceramics, although the HA layer on the AOFCP was thin at 1-2 microns. From SEM observation of the pushed-out specimen, it was found that the thin HA layer had directly bonded to bone but the AOFCP had not. The push-out strength of the hydrothermally treated film resulted from the chemical bonding of the bone-HA layer interface, while that of the untreated film resulted from mechanical interlocking force between bone and the microprojections. There was a small difference in bone apposition but no significant difference in push-out strength with the amount of precipitated HA crystals on the treated films. Among the treated films, the film formed at the lowest electrolyte concentration showed the lowest bone apposition because of incomplete covering by the HA crystals, and showed the highest stability against mechanical failure because the adhesive strength was very high at about 38 mPa. Also, the hydrothermally untreated anodic oxide films, whose surfaces were rough as a result of the large microprojections, showed much higher push-out strength and bone apposition than titanium. The good hard-tissue compatibility may be attributed to the surface roughness and the possible inhibition of titanium ion release from the specimen.

Characterization of thin hydroxyapatite layers formed on anodic titanium oxide films containing Ca and P by hydrothermal treatment.

An anodic titanium oxide film containing Ca and P (AOFCP) was formed on commercially pure titanium which was anodized in an electrolytic solution of dissolved beta-glycerophosphate (beta-GP) and calcium acetate (CA). Hydroxyapatite (HA) crystals were precipitated by hydrothermally heating the AOFCP at 300 degrees C. After hydrothermal treatment, the film was characterized by scanning electron microscopy (SEM), energy-dispersive X-ray microanalysis (EDX), and tensile tests. The morphology, composition, and amount of HA crystals precipitated were significantly affected by the composition of the electrolytes. Near-stoichiometric HA crystals with high crystallinity were precipitated completely covering the AOFCP surface at specific electrolyte concentrations. The HA layers were thin at 1-2 microns in thickness. The adhesive strength of the film increased with decreasing electrolyte concentration and the maximum value was about 40 MPa. In vitro tests for 300 days suggested that the stability of the film was high. The high adhesive strength may result from the AOFCP existing as an intermediate layer between the HA layer and a titanium substrate. The intervention of the AOFCP may have prevented abrupt changes in Ca and P content at an HA coating-titanium interface as seen in a plasma-sprayed one. The porous TiO2 matrix of the AOFCP may be suitable for nucleation sites of HA crystals, as well as SiO2 matrix of silicate bioactive glasses or glass ceramics.

Formation and characterization of anodic titanium oxide films containing Ca and P.

Commercially pure titanium was anodized in an electrolytic solution that was dissolved calcium and phosphorus compounds in water, and an AOFCP (anodic titanium oxide film containing Ca and P) was formed. It was found that sodium beta-glycerophosphate (beta-GP) and calcium acetate (CA) were suitable for the electrolytes to form the AOFCP having an equivalent Ca/P ratio to hydroxyapatite (HA). The AOFCP was characterized by scanning electron microscopy (SEM), an energy-dispersive X-ray microanalysis (EDX), and X-ray diffraction (XRD). Numerous micropores and microprojections were observed on the AOFCP by SEM. The composition of the AOFCP, which was measured by EDX, changed according to beta-GP and CA concentration, and the electrolytic voltage. Ca and P in the AOFCP seem to be incorporated into the TiO2 matrix from CA and beta-GP in the electrolyte during the anodic oxidation. Despite the existence of Ca and P in the AOFCP, no calcium phosphate peak was detected by XRD, and the AOFCP consisted of anatase and only a little rutile. The AOFCP, whose contents of Ca and P were low, had a high adhesive strength after soaking in a simulated body fluid for 300 days. When the AOFCP having an equivalent Ca/P ratio to HA was hydrothermally heated at 300 degrees C, HA crystals were precipitated on the AOFCP and completely covered the surface.

Hypothalamic vasopressin mRNA levels in mice are decreased after chronic ethanol ingestion.

Vasopressin mRNA levels in the supraoptic and paraventricular nuclei of the hypothalamus, measured by in situ hybridization with a 35S-labeled RNA probe, were decreased by nearly 50% in C57BL/6NCR mice that had ingested an ethanol-containing diet for 7 days, and were tolerant to and physically dependent on ethanol. At 24 h after withdrawal, vasopressin mRNA levels in the supraoptic nucleus were still decreased, while levels in the paraventricular nucleus returned toward control values. Although plasma osmolality was increased in ethanol-fed mice, there was no increase in plasma vasopressin levels, possibly as a result of the effect of chronic ethanol ingestion to decrease vasopressin synthesis. In contrast, in mice that were dehydrated, but not fed ethanol, plasma osmolality, plasma vasopressin levels, and hypothalamic vasopressin mRNA all increased, as expected. The data suggest that chronic ethanol ingestion interferes with the synthesis and secretion of vasopressin, and may result in the reduced ability of an individual to respond to physiological stimuli for vasopressin secretion.

Oligonucleotidase activity of phosphodiesterase from the fruit body of Flammulina velutipes.

A phosphodiesterase (EC 3.1.4.1) was purified to homogeneity from the fruit body of Flammulina velutipes. The enzyme had considerable activity toward oligonucleotides. The Km values were 0.66 mM for ApA, 2.47 mM for (Ap)2A, and 3.03 mM for (Ap)3A. The enzyme hydrolyzed oligodeoxyribonucleotides as well as oligoribonucleotides. The oligoribonucleotides bearing a phosphate residue at the 3' end were not hydrolyzed by the enzyme. The enzyme hydrolyzed the oligoribonucleotides exonucleolytically from the 3' to 5' end. Thus the PDase of F. velutipes is considered to function in vivo as an oligonucleotidase (EC 3.1.13.3), which efficiently converts oligonucleotides to 5'-mononucleotides in the cell.

The role of arginine vasopressin in alcohol tolerance.

Administration of the neuropeptide, arginine vasopressin, to animals that have acquired functional tolerance to ethanol will maintain such tolerance, even in the absence of further ethanol ingestion by the animals. In mice, this action of the peptide is mediated by central nervous system V1 receptors and requires intact brain noradrenergic systems. Autoradiographic studies have shown that some V1 receptors are localized presynaptically on catecholaminergic neuronal terminals in the mouse lateral septum, suggesting that vasopressin may act via modulation of catecholamine release. In addition, vasopressin has been found to increase mRNA levels for the proto-oncogene, c-fos, in septum and hippocampus, possibly by an action at postsynaptic receptors. Expression of c-fos, which has been hypothesized to play a role in central nervous system neuroadaptation, could transform short-term actions of vasopressin into long-term effects on ethanol tolerance. Studies with vasopressin antagonists indicate that the endogenous peptide influences tolerance, and therefore the effect of chronic ethanol ingestion on vasopressin synthesis and release was studied. In mice and rats, hypothalamic vasopressin mRNA is decreased by chronic ethanol exposure, although effects on plasma vasopressin levels differ in the two species. The effect of ethanol on extrahypothalamic vasopressin synthesis in brain is under investigation. The results suggest mechanisms by which vasopressin can produce long-term changes in central nervous system function, and provide evidence for a disturbance of vasopressin regulation during chronic ethanol ingestion.

Reduction of arginine vasopressin binding sites in mouse lateral septum by treatment with 6-hydroxydopamine.

The neuropeptide arginine vasopressin modulates neuroadaptive processes, including memory consolidation and functional tolerance to ethanol, by actions at CNS V1 receptors. Noradrenergic systems play a role in these actions of the peptide. To assess whether vasopressin may act presynaptically on catecholamine neurons, vasopressin receptors were measured by quantitative autoradiography in the lateral septum, an area that is innervated by catecholaminergic neurons and has a high density of V1 receptors, of control and 6-hydroxydopamine-treated mice. Vasopressin receptors were distributed non-uniformly throughout the lateral septum, with greater binding in the more caudal regions. Treatment with 6-hydroxydopamine lowered septal catecholamine levels and vasopressin binding, with a greater effect on binding in the intermediate and caudal portions of the lateral septum. Pretreatment with desmethylimipramine reversed the depletion of norepinephrine, and attenuated the effect of 6-hydroxydopamine on vasopressin binding in the intermediate region, but was less effective in the caudal region of the lateral septum. The results suggest that a portion of septal vasopressin receptors are localized on the terminals of noradrenergic and, possibly, dopaminergic neurons, consistent with the hypothesis that certain neuroadaptive responses to vasopressin could be mediated by modulation of neurotransmitter release. In contrast to the results with 6-hydroxydopamine, treatment of mice with 5,7-dihydroxytryptamine, to destroy serotonergic terminals, did not alter vasopressin binding in the lateral septum.

Effects of ethanol on adenylate cyclase system in the human platelet.

Effects of ethanol (EtOH) on the alpha 2-receptor-coupled adenylate cyclase (AC) of the human platelet were examined. EtOH increased "basal" AC activity in a linear dose dependent manner. In the presence of Gpp(NH)p (2.5 microM), however, the slope of the dose-response curve for EtOH stimulation of AC activity was biphasic. The increase in activity produced by the addition of EtOH concentration between 0 and 100 mM was much sharper than the increase in activity produced by the concentration in excess of 100 mM. EtOH increased the rate of activation of AC by guanine nucleotides and concomitantly decreased the concentration of magnesium required for half-maximal activation of AC. Prostaglandin-E1 (PGE1) alone stimulated AC activity. Clonidine (3 nM-1 microM) diminished the PGE1 (1 microM)-stimulated AC by a maximum of 30%. EtOH did not alter the concentration of clonidine required for half-maximal inhibition of AC. Our results suggest that the sites of EtOH's action can be assigned to the direct action of the catalytic subunit and to the Ns-protein. Our results also indicate a substantial difference in EtOH's action on catecholamine receptor systems coupled in a stimulatory versus inhibitory manner to AC. Stimulation of AC through Ns-protein is potentiated by EtOH, but inhibition of AC through the Ni-protein is little affected by EtOH.

Neurochemical findings in the cerebrospinal fluid of schizophrenic patients with tardive dyskinesia and neuroleptic-induced parkinsonism.

Monoamine and their acid metabolites were determined in the CSF of 18 drug-treated chronic schizophrenic patients with the symptoms of tardive dyskinesia and neuroleptic-induced Parkinsonism (Parkinsonism). Six healthy volunteers were used as the control group. The norepinephrine (NE) levels were found to be significantly higher in the patients with tardive dyskinesia than in the controls. Furthermore, elevated CSF NE levels were also observed in the patients with Parkinsonism. Epinephrine (E) and Dopamine (DA) were not present in the CSF of the control group, whereas measurable levels of DA could be detected in 4 out of 9 and E was found in 8 out of 9 patients with tardive dyskinesia. The mean concentration of HVA was slightly but not significantly elevated in the patients with tardive dyskinesia and Parkinsonism. The mean values of CSF 5-HIAA were all within the normal range in both patient groups. From the above results, it was suggested that abnormal adrenergic activity rather than abnormal dopaminergic activity may play an important role as a mechanism in the etiopathogenesis of extra-pyramidal disorders. Furthermore, in the patients with Parkinsonism, CSF neurochemical observations were similar to those of the patients with tardive dyskinesia in this study. It may help to explain the clinical coexistence of tardive dyskinesia and neuroleptic-induced Parkinsonism.

Age-associated alterations in hepatic glutathione-S-transferase activities.

Age-associated alterations of hepatic cytosolic glutathione-S-transferase (GST) activities towards sulfobromophthalein sodium tetrahydrate (BSP), styrene oxide (STOX), trans-4-phenyl-3-butene-2-one (PBO), 1,2-dichloro-4-nitrobenzene (DCNB), and 1-chloro-2,4-dinitrobenzene (CDNB) were investigated in Fischer-344 rats of both sexes with ages ranging from 1.5 to 28 months. The GST activities towards PBO and DCNB in male rats increased with age till 6-12 months when maximum values were attained, and then gradually decreased till 28 months when the values became the lowest. The GST activities towards STOX and BSP did not show any significant increase after 1.5 months and stayed at this level till 12 months, followed by a gradual decrease till 28 months when the values were the lowest. In contrast, the GST activity towards CDNB in male rats did not show much of an age-associated alteration. Age-associated alterations in GST activities in females were much smaller than those observed in males. Sex differences in GST activities (significantly higher male values than female values) were observed with all the substrates examined at least at some time of the animal life. The kinetic studies of GST activities indicated that alterations in the relative abundance as well as the total quantity of GST isozymes caused the substrate selective alterations of GST activities with age.

Pig skin epidermal calmodulin: effect on calmodulin deficient phosphodiesterase.

Calmodulin, a calcium-dependent modulator protein, is known to mediate a great number of Ca++-dependent processes in various tissues. Although it was originally described as a protein activator of cyclic nucleotide phosphodiesterase, the sensitivity of phosphodiesterases to this compound are suggested to be variable from tissue to tissue. In order to determine whether there was calmodulin-like activity in pig skin epidermis and to see its relationship to epidermal phosphodiesterase, we used an established calmodulin deficient phosphodiesterase system prepared from bovine heart. Calmodulin deficient phosphodiesterase prepared from bovine heart was markedly stimulated by the addition of pig skin (epidermal) boiled extract in the presence of calcium. Boiled skin extract alone had only little phosphodiesterase activity by itself. This effect of boiled skin extract on bovine heart phosphodiesterase was inhibited by the addition of EGTA, a divalent metal ion chelator of relative Ca++ specificity. At a fixed concentration of EGTA, increasing the Ca++ concentration counteracted the effect of EGTA. Pure pig skin epidermis (separated by trypsinization, NaBr, CaCl2-sucrose or NH4Cl treatment) was also shown to have heat-stable calmodulin activity. In contrast to the bovine heart phosphodiesterase, epidermal phosphodiesterase was only partially inhibited when Ca++ was removed by EGTA. The addition of boiled skin extract on the crude extract of epidermal phosphodiesterase had minimal effect on the enzyme activity. Overall results indicate that although pig skin epidermis contains significant amount of calmodulin, the regulation of phosphodiesterase may not be the main biological activity of epidermal calmodulin.