Growth and photosynthetic responses of two pine species (Pinus koraiensis and Pinus rigida) in a polluted industrial region in Korea.

We investigated the effects of pollutants on two pine species (Pinus koraiensis and Pinus rigida) in an industrial region in Korea, using a physiological approach. The concentrations of fluorine (F) and chlorine (Cl) in the atmosphere, in precipitation and soil water at the damaged site were all significantly higher than at a control site. Moreover, the concentrations of F, Cl and Mn in pine needles were significantly higher, and essential elements and chlorophyll in needles were significantly lower at the damaged site than at the control site. The photosynthetic capacities, shoot length and survival statistics of needles of the two pines were all significantly reduced at the damaged site compared to the control site, especially P. rigida. Based on our comparison of photosynthetic responses and the concentrations of F, Cl and Mn in needles of the two pine species, P. koraiensis is more resistant to excess Mn in its needles than P. rigida.

Biochemical and pharmacological characterization of YM-60828, a newly synthesized and orally active inhibitor of human factor Xa.

YM-60828 was found to potently inhibit human factor Xa following oral administration. YM-60828 showed high affinity for factor Xa (Ki = 1.3 nM), but did not affect thrombin (Ki > 100 microM). YM-60828 doubled factor Xa clotting time, prothrombin time (PT) and activated partial thromboplastin time (APTT) at 0.10, 0.21, 0.24 microM, respectively. Importantly, it did not prolong thrombin time at 100 microM. YM-60828 also inhibited factor Xa in the prothrombinase complex with an IC50 value of 7.7 nM. In addition to its anticoagulant activity, YM-60828 inhibited platelet aggregation induced by various agonists (IC50 = 3 to 23 microM). Squirrel monkeys were used to study the ex vivo anticoagulant activity and pharmacokinetic properties of YM-60828. One hour after oral administration at 3 mg/kg, YM-60828 strongly prolonged PT and APTT by 4.8- and 1.9-fold, respectively, and plasma concentration reached 788 +/- 167 ng/ml. Bioavailability was calculated to be 20.3%. These results strongly suggest that YM-60828 will be a valuable orally active and potent anticoagulant agent showing potential antithrombotic activity.

Hydrolysis and synthesis of wax esters by different systems of carp hepatopancreas preparation.

It was found that the supernatant of hepatopancreas homogenate of the carp, Cyprinus carpio, contained enzymes involved in the hydrolysis and esterification of wax esters. The enzymes were salted out by 30% saturation with ammonium sulfate. This fraction also contained the highest activities of lipase and esterase. The enzyme for the hydrolysis of wax esters, together with lipase and esterase, was eluted in tube numbers 20 to 22 (void volume) from a Sephadex G-200 column. On the other hand, the enzyme for the synthesis of wax esters was eluted in tube numbers 33 to 35. It thus appears that the two activities belong to different enzyme proteins. The synthetic enzyme was activated by the addition of ATP, CoA, and NADH.

Analysis of thromboxane B3 converted from eicosapentaenoic acid in human platelet rich plasma by gas chromatography/mass spectrometry.

Gas chromatographic mass spectrometric determination of thromboxane B3 (TXB3) synthesized from platelet is described. Eicosapentaenoic acid (EPA) was added to human platelet rich plasma and after the reaction the exstraction was carried out. Plasma thromboxanes were run through an Amberlite XAD-2 and SEP-PAK silica cartridge, and then chromatographed using silicagel thin layer plate to remove interfering materials, such as 6-keto-prostaglandin F1 alpha. Extracted thromboxanes were converted into the methoxime-dimethylisopropylsilylmethyl ester derivatives and they were measured by gas chromatography/ammonia chemical ionization mass spectrometry. Three peaks were obtained on the gas chromatogram which were presumed to be 3-series metabolite product TXB3 and their related substances. Results indicates the human platelet may easily convert EPA to TXB3 by adding EPA to PRP without adding arachidonic acid.

Deregulated production of interleukin-4 (IL4) in autoimmune thyroid disease assayed with a new radioimmunoassay.

A sensitive, reproducible and specific radioimmunoassay for human interleukin-4 (IL4) has been developed. Using 125I-labeled IL4 and polyclonal rabbit antisera raised against recombinant human IL4, a competitive inhibition assay was developed which could detect 5 pg/ml of human IL4. Other interleukins, growth factors, hormones, peptides and lectins did not affect the assay. IL4 was measured in supernatants of culture media of stimulated human peripheral blood mononuclear cells (PBMC). Kinetics of IL4 production in PHA-stimulated PBMC from seven normal subjects revealed that the peak levels of IL4 were seen at 24 h and then declined. Peak IL4 levels in PHA stimulation of PBMC from untreated patients with autoimmune thyroid diseases (Graves' disease and chronic thyroiditis) were significantly higher than normal controls. However, after treatment, IL4 production decreased to normal. The present study demonstrates the usefulness of quantitating human IL4 produced by PBMC and that there exists a deregulated production of IL4 in autoimmune thyroid diseases.

Radioimmunoassay for orexin A.

A radioimmunoassay for orexin A has been developed. Anti-orexin A antiserum was raised in New Zealand white rabbits immunized with a conjugate of synthetic orexin A with bovine serum albumin. This antibody did not crossreact with orexin B, hypothalamic hormones, pituitary hormones, neuropeptides or gut hormones. Radioiodination of orexin A was performed with the chloramin T method, followed by purification of radioiodinated material on Sephadex G-25 column. Orexin A was extracted from tissues using acid-acetone. The assay was performed with a double antibody system. The dilution curve of acid-acetone-extracts of rat hypothalamus in the radioimmunoassay system was parallel to the standard curve. The recovery of tissue orexin A was about 80%,and the intra-assay and inter-assay variations were 5.2% and 7.8%, respectively. Orexin A was found in the hypothalamus, cerebrum and testis. These data suggest that this assay system is suitable for the measurement of tissue orexin A and that orexin A is found in the central nervous system and testis.

Thyrotropin-releasing hormone and somatostatin inhibit each others release in vitro in the rat retina.

The effect of thyrotropin-releasing hormone (TRH), somatostatin (SS) or octreotide, an analogue of SS, on release of TRH or SS from the rat retina was studied in vitro. The retina was incubated in medium 199 (pH 7.4) with 1.0 mg/ml of bacitracin (medium) for 20 min. The amount of TRH or SS release into the medium was measured by individual radioimmunoassays. The TRH release from the rat retina was inhibited significantly in a dose-related manner by the addition of SS or octreotide. The SS release from the retina was inhibited by TRH, and the inhibitory effect of TRH on SS release from the rat retina was blocked by the addition of anti-TRH receptor antiserum immunoglobulin fraction. The findings suggest an interaction between TRH and SS in the rat retina by which the addition of one inhibits the release of the other.

Growth and annual ring structure of Larix sibirica grown at different carbon dioxide concentrations and nutrient supply rates.

We compared effects of ambient (360 vpm) and elevated (720 vpm) carbon dioxide concentration ([CO2]) and high and low nutrient supply rates on stem growth, annual ring structure and tracheid anatomy of Siberian larch (Larix sibirica Ledeb.) seedlings over two growing seasons. Elevated [CO2] had no significant effect on either stem height or diameter growth; however, both stem height and diameter growth were enhanced by the high nutrient supply rate, and these increases were stimulated by elevated [CO2]. Elevated [CO2] tended to increase the width of the annual xylem ring, the number of cells in a radial file spanning the ring, and tracheid lumen diameter, whereas it tended to reduce cell wall thickness, although there were no statistically significant CO2 effects on tracheid anatomy. Changes in tracheid cell morphology seemed to be dependent on changes in shoot elongation rates.

Effects of deicing salt on the vitality and health of two spruce species, Picea abies Karst., and Picea glehnii Masters planted along roadsides in northern Japan.

In northern Japan, the growth of Picea abies Karst., and Picea glehnii Masters, which have been planted along the highways, is often suppressed due to several environmental stresses. To examine the adverse effects of deicing salt, the primary source of stress,we measured needle life span, photosynthetic capacity, and water potential and transpiration rate of the two spruce species at a site with damaged trees, near the roadside and a site with healthy trees, located far from the highway. Results from the analysis showed large amounts of sodium and chlorine in the soil and snow at the damaged site. These elements had accumulated in the needles of the spruce. Moreover, physiological traits of the spruce, at the damaged site were also affected. Therefore, we concluded that poor physiological traits might be attributed to an accumulation of deicing salt in the needles, resulting in the suppression of tree growth.

Metabolism of orally administered ergosterol and 7-dehydrocholesterol in rats and lack of evidence for their vitamin D biological activity.

Metabolism of orally administered ergosterol (Erg) and 7-dehydrocholesterol (7-DHC) in rats and their vitamin D biological activity were investigated. Most of orally administered Erg and 7-DHC were excreted in feces and the remaining sterols were absorbed through intestine. The absorbed sterols were not transported in skin as the intact forms but metabolized into brassicasterol and cholesterol, respectively, within 25 h. Neither increment of intestinal calcium absorption nor plasma calcium concentrations were observed by oral administration of Erg and 7-DHC to vitamin D-deficient rats. Therefore, we have concluded that orally administered Erg and 7-DHC have no vitamin D biological activity.

Japanese expert public health nurses empower clients with schizophrenia living in the community.

1. Home visiting psychiatric public health nurses are integral to expanding community-based mental health services for Japanese citizens living with schizophrenia and other mental illnesses. 2. Interviews with experienced psychiatric public health nurses revealed empowerment as the basic social process that guides their practice. Five empowering practice domains foster client autonomy. 3. Psychiatric public health nurses work with families, neighbors, educators, and employers to enable their clients' healthy living in the community.

[Effectiveness of home help service for psychiatric patients. The results of a model project in S city].

PURPOSE: The purpose of this study was to evaluate the effectiveness of a home help service project for psychiatric patients in S city. METHODS: To assess the effectiveness of the home help service, home help service providers at S city in 1997 were invited to hearings and the transcripts were examined qualitatively. Based on the results, interview questions were developed. We then interviewed 28 home help providers who were providing or had completed the service project, as well as care managers as of July 1998. Home help providers divided their patient stories retrospectively into three time frames: before starting home care, one month after starting providing care and at the present. Amount of care provided and patients' self care level were scored and averages ere compared for the three time frames. RESULTS: The analysis of the hearing data revealed parameters for the effectiveness of home help service for psychiatric patients to be as follows: 1) enhanced ability of daily living, such as taking meals, maintaining hygiene and shopping, (primary effectiveness: 5 items); and 2) developing social function, such as being motivated, maintaining personal relationship, extending social life, and decreasing family burden, (secondary effectiveness: 9 items). The amount of care needed was found to be significantly decreased and social function was significantly enhanced on comparisons between before starting home care and one month later, as well as between before starting home care and the present. DISCUSSION: Home help service for psychiatric patients is not only useful for enhancing patients' ability to succeed in daily living tasks, but also for developing their social functioning. Effectiveness was also identified in the level of patients' behavior. The results provide reasonable grounds for extended use of the home help service.

Programmed necrosis, not apoptosis, is a key mediator of cell loss and DAMP-mediated inflammation in dsRNA-induced retinal degeneration.

There is no known treatment for the dry form of an age-related macular degeneration (AMD). Cell death and inflammation are important biological processes thought to have central role in AMD. Here we show that receptor-interacting protein (RIP) kinase mediates necrosis and enhances inflammation in a mouse model of retinal degeneration induced by dsRNA, a component of drusen in AMD. In contrast to photoreceptor-induced apoptosis, subretinal injection of the dsRNA analog poly(I : C) caused necrosis of the retinal pigment epithelium (RPE), as well as macrophage infiltration into the outer retinas. In Rip3(-/-) mice, both necrosis and inflammation were prevented, providing substantial protection against poly(I : C)-induced retinal degeneration. Moreover, after poly(I : C) injection, Rip3(-/-) mice displayed decreased levels of pro-inflammatory cytokines (such as TNF-α and IL-6) in the retina, and attenuated intravitreal release of high-mobility group box-1 (HMGB1), a major damage-associated molecular pattern (DAMP). In vitro, poly(I : C)-induced necrosis were inhibited in Rip3-deficient RPE cells, which in turn suppressed HMGB1 release and dampened TNF-α and IL-6 induction evoked by necrotic supernatants. On the other hand, Rip3 deficiency did not modulate directly TNF-α and IL-6 production after poly(I : C) stimulation in RPE cells or macrophages. Therefore, programmed necrosis is crucial in dsRNA-induced retinal degeneration and may promote inflammation by regulating the release of intracellular DAMPs, suggesting novel therapeutic targets for diseases such as AMD.

The prostaglandin synthesis in marine fish thrombocyte.

1. Prostaglandin (PG) syntheses from labelled highly unsaturated fatty acids were investigated in washed thrombocyte suspensions of four species of marine fish, flounder (Paralichthys olivaceus), black seabream (Acanthopagrus schlegeli), black rockfish (Sebastes schlegeli), and red seabream (Pagrus major). 2. Synthesized PGs were analyzed by thin-layer radiochromatogram scanner and high-performance liquid chromatography. 3. When [1-(14)C]arachidonic acid (AA) was incubated with washer thrombocyte suspension, AA was mainly converted to PGF(2alpha), PGE2, and PGD2 in all species. 4. It was suggested that [1-(14)C]eicosapentaenoic acid was mainly converted in PGs only in black rockfish thrombocytes, and the metabolites were mainly PGE3 and PGD3. 5 However, [1-(14)C]docosahexaenoic acid was not cyclized into PGs in all the species.

Effects of acetylcholine on the release of thyrotropin-releasing hormone from the rat retina in vitro.

Effects of acetylcholine on the release of thyrotropin-releasing hormone (TRH) from the rat retina were studied in vitro. The retina was incubated in medium 199 (pH 7.4) with 1.0 mg/ml of bacitracin and 100 micrograms/ml of ascorbic acid (medium) for 20 min. The amount of TRH release into the medium was measured by radioimmunoassay. The TRH release from the rat retina was enhanced significantly in a dose-related manner with the addition of acetylcholine and inhibited with addition of atropine. The stimulatory effect of acetylcholine on TRH release from the retina was blocked with the addition of atropine. The elution profile of methanol-extract of the rat retina was identical to that of synthetic TRH. The findings suggest that the cholinergic system stimulates TRH release from the rat stomach in vitro.

Purification and gas chromatographic-mass spectrometric characterization of non-methylene interrupted fatty acid incorporated in rat liver.

A C20 non-methylene interrupted trienoic acid detected in the liver of rat fed with a pine (Pinus koraiensis) seed oil diet was purified by two-step argentation thin-layer chromatography (AgTLC) and characterized by gas chromatography-mass spectrometry (GC-MS). First, a C20 methyl trienoate fraction was obtained from fatty acid methyl esters prepared from rat liver by 5% AgTLC developed with petroleum ether-diethyl ether-acetic acid (70:20:2, v/v) as a solvent system. The fraction was then subjected to AgTLC developed with benzene-acetone-diethyl ether-acetic acid (65:15:15:5, v/v) which could separate non-methylene interrupted fatty acids (NMIFA) from usual MIFAs. The purified C20 NMIFA was partially hydrogenated, and the resulting three kinds of the C20 monoenoate were analyzed by GC-MS after conversion to their dimethyl disulfide (DMDS) adducts. The results revealed that the original C20 non-methylene interrupted trienoic acid detected in the liver of rats fed with a pine seed oil diet was delta-5,11,14/20:3, a minor component of pine seed oil.