Effects of delta-opioid receptor stimulation and inhibition on hippocampal survival in a rat model of forebrain ischaemia.

BACKGROUND: It has been reported that delta-opioid (DOP) receptor agonists may be neuroprotective in the central nervous system. However, the DOP agonist [d-Ala(2), d-Leu(5)]enkephalin (DADLE) does not produce neuroprotection in severe forebrain ischaemia. The aim of this study was to examine the effects of DADLE on hippocampal neurone survival against less severe forebrain ischaemia. METHODS: Intraperitoneal injection of DADLE (0 or 16 mg kg(-1)) in male Sprague-Dawley rats was performed 30 min before ischaemia. Severe (10 min), moderate (8 min), or mild (6 min) forebrain ischaemia was produced by bilateral carotid occlusion combined with hypotension (35 mm Hg) under isoflurane (1.5%) anaesthesia. Naltrindole (10 mg kg(-1)) (DOP antagonist) was administered 30 min before DADLE in order to confirm DOP receptor activation in the neuroprotective efficacy of DADLE. Naltrindole alone was also administered 30 min before ischaemia to examine endogenous DOP agonism as a self-protecting mechanism against ischaemia. All animals were evaluated neurologically and histologically after a 1 week recovery period. RESULTS: DADLE improved neurone survival in hippocampal CA3 and dentate gyrus (DG) sectors. CA1 neurones were not protected against moderate and mild ischaemia. Naltrindole abolished DADLE neuroprotection in the CA3 and DG after both moderate and mild ischaemia. Interestingly, regardless of co-administration of DADLE, naltrindole significantly worsened neuronal injury in the CA1 region after mild ischaemia. CONCLUSIONS: These results suggest that DADLE provides limited neuroprotection to relatively ischaemia-resistant regions but not to selectively vulnerable regions. This was probably mediated by DOP stimulation. Pre-ischaemic treatment with a DOP antagonist, regardless of co-administration of DADLE, worsened neuronal damage at the selectively vulnerable regions only after mild forebrain ischaemia. These data suggest that DOP activation with endogenous DOP ligand may be involved in self-protecting ischaemia-sensitive regions of the brain.

Toxicological study of a new maintenance fluid, Veen 3G, in rats.

A study of the different volume and infusion rates of a new maintenance fluid, Veen 3G, on the general conditions of rats was investigated during the 14 days after infusion. In Experiment I, 100 ml/kg and 200 ml/kg of Veen 3G were infused at a rate of 300 ml/kg/h in male and female rats. Results were compared with those for Gurunon Ringer solution (GRS) in male and female rats. We observed only transient polyuria in animals administered by each dose of Veen 3G and GRS for 0-15 min after infusion. Necropsy was not observed in any of the animals tested 14 days after infusion. In Experiment II, 200 ml/kg of Veen 3G was infused at rates of 200, 400, 800 and 1600 ml/kg/h in male rats. At 800 and 1600 ml/kg/h, irregular respiration and decrease in movement were observed concomitantly with polyuria. Three out of 4 rats died immediately after the infusion of Veen 3G at a rate of 1600 ml/kg/h, and one rat was still alive 14 days after the infusion. In this experiment, 200 ml/kg Veen 3G was safe when we infused at a rate of less than 400 ml/kg/h in male rats. Since this rate is about 27-80 times higher than that used clinically in maintenance treatment, Veen 3G is suggested to be safe, with the exception of polyuria, in clinical situations at the standard infusion rate (5-15 ml/kg/h).

Identification of a novel cytokine, ML-1, and its expression in subjects with asthma.

A novel gene, designated ML-1, was identified from a human genomic DNA clone and human T cell cDNA sequences. The second exon of ML-1 gene shares significant sequence identity with the gene encoding IL-17 (IL-17). ML-1 gene expression was up-regulated in activated PBMCs, CD4(+) T cells, allergen-specific Th0, Th1, and Th2 clones, activated basophils, and mast cells. Increased expression of the ML-1 gene, but not IL-17, was seen following allergen challenge in four asthmatic subjects, suggesting its role in allergic inflammatory responses. ML-1 from transiently transfected COS-7 cells was able to induce gene expression and protein production for IL-6 and IL-8 (at 10 ng/ml of ML-1: for IL-6, 599.6 +/- 19.1 pg/ml; for IL-8, 1724.2 +/- 132.9 pg/ml; and at 100 ng/ml of ML-1: for IL-6, 1005.3 +/- 55.6 pg/ml; for IL-8, 4371.4 +/- 280.5 pg/ml; p < 0.05 for both doses vs baseline) in primary bronchial epithelial (PBE) cells. Furthermore, increased expression of ICAM-1 was found in ML-1-stimulated PBE cells (mean fluorescence intensity (MFI) = 31.42 +/- 4.39 vs baseline, MFI = 12.26 +/- 1.77, p < 0.05), a functional feature distinct from IL-17 (MFI = 11.07 +/- 1.22). This effect was not inhibited by a saturating amount of IL-17. These findings demonstrate that ML-1 is a novel cytokine with a distinct function, and suggest a different receptor for ML-1 on PBE cells.

Non-Hodgkin's lymphoma of the ascending colon in a patient with becker muscular dystrophy: report of a case.

We herein present the findings of a 10-year-old boy with non-Hodgkin's lymphoma of the ascending colon which caused intussusception and intestinal bleeding. He had a history of Becker muscular dystrophy. However, he had neither hypertrophic calves nor cardiomyopathy, and his serum creatine kinase (CK) level always exceeded 2000 IU/l. Preoperatively, a laboratory examination revealed high serum levels of CK (2038IU/l), aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH), and the blood hemoglobin level was 7.0g/dl. A barium enema examination revealed an intussusception in his ascending colon, which was found to be a highly vascular tumor on Doppler ultrasound scans. A right hemicolectomy was performed. Macroscopically, the 5 x 6 x 8-cm solid tumor of the ascending colon resembled a submucosal tumor and had two ulcerous lesions at the tip. The tumor was histologically diagnosed to be a diffuse large B-cell lymphoma of the ascending colon. General examinations revealed no involvement of lymphoma postoperatively. At 13 months after surgery, the CK (37861U/l), AST (110lU/l), ALT (1381U/ l), and LDH (420lU/l) levels are still high, and the patient is doing well without any signs of recurrence.

Analysis of ENOD40 expression in alb1, a symbiotic mutant of Lotus japonicus that forms empty nodules with incompletely developed nodule vascular bundles.

The alb1 mutant of Lotus japonicus (Ljsym74) forms empty nodules in which most of the bacteria remain in abnormally enlarged infection threads and fail to enter the host plant cells. The alb1 mutant was also found to be defective in differentiation of ramified nodule vascular bundles; only a single vascular bundle differentiates at the proximal end of the alb1 nodules and it fails to differentiate further. Histochemical analysis using fluorescein-conjugated wheat-germ agglutinin (F-WGA) indicated that the mutation in the ALB1 gene specifically affects the differentiation of vascular bundles in nodules. Analysis of nodulin gene expression revealed that the expression of an early nodulin gene, ENOD40, was very low in alb1 nodules. At early developmental stages of alb1 nodules, the pattern of ENOD40 transcription was essentially the same as that in wild-type nodules; transcripts were localized in dividing cortical cells and in the pericycle of the root stele opposite nodule primordia, as in wild-type nodules. However, mature alb1 nodules exhibited very weak or no expression of ENOD40 in the peripheral cells of the undeveloped nodule vascular bundle. The ENOD40 expression pattern in alb1 nodules is distinct from that in another ineffective mutant, fen1 (Ljsym76), in which ENOD40 expression persists prior to premature senescence. These findings lead us to speculate that ENOD40 may play a role in the differentiation of nodule vascular bundles.

Isolation of two different phenotypes of mycorrhizal mutants in the model legume plant Lotus japonicus after EMS-treatment.

Lotus japonicus has been proposed as a model plant for the molecular genetic study of plant-microbe interaction including Mesorhizobium loti and arbuscular mycorrhizal (AM) fungi. Non-mycorrhizal mutants of Lotus japonicus were screened from a collection of 12 mutants showing non-nodulating (Nod-), ineffectively nodulating (Fix-) and hypernodulating (Nod++) phenotypes with monogenic recessive inheritance induced by EMS (ethylmethane sulfonate) mutagenesis. Three mycorrhizal mutant lines showing highly reduced arbuscular mycorrhizal colonization were obtained. All of them were derived from Nod- phenotypes. In Ljsym72, the root colonization by Glomus sp. R-10 is characterized by poor development of the external mycelium, formation of extremely branched appressoria, and the blocking of hyphal penetration at the root epidermis. Neither arbuscules nor vesicles were formed in Ljsym72 roots. Fungal recognition on the root surface was strongly affected by the mutation in the LjSym72 gene. Unique characteristics in mutant lines Ljsym71-1 and Ljsym71-2 were the overproduction of deformed appressoria and arrested hyphal penetration of the exodermis. Small amounts of internal colonization including degenerated arbuscule formation occurred infrequently in these types of mutants. Not only fungal development on the root surface but also that in the root exodermis and cortex was affected by the mutation in LjSym71 gene. These mutants represent a key advance in molecular research on the AM symbiosis.

The mouse ZFH-4 protein contains four homeodomains and twenty-two zinc fingers.

We isolated the mouse zfh-4 cDNA which is 12 kb long and capable of encoding a 3,550-amino acid protein containing four homeodomains and 22 zinc fingers including two pseudo zinc finger motifs. The mouse ZFH-4 is 51% homologous to the mouse ATBF1 and 23% to the Drosophila ZFH-2. The homeodomain and zinc finger regions are highly conserved between ZFH-4 and ATBF1 except that one zinc finger is missing in ZFH-4. Analysis of partial genomic sequences showed that the mouse zfh-4 and ATBF1 genes are similar in exon-intron organization. RT-PCR analysis of zfh-4 transcripts in adult mouse tissues showed that zfh-4 expression was low but reproducibly detectable in brain, heart, lung and muscle. In these mouse tissues, ATBF1 transcripts were poorly amplified by PCR under the conditions where zfh-4 transcripts were amplified, suggesting that the expression of zfh-4 mRNA is higher than that of ATBF1 mRNA. Other comparative analysis suggests functional similarities and dissimilarities between ZFH-4 and ATBF1.

Clinical applicability of functional magnetic resonance imaging and activation study with single photon emission computerized tomography for functional mapping.

The purpose of this study is to estimate the clinical applicability of functional magnetic resonance imaging (f-MRI) combined with the activation study using single photon emission computerized tomography (SPECT) with special reference to identification of primary sensorimotor area. Five healthy volunteers and 5 patients with brain tumors located around the sensorimotor cortex were studied by both f-MRI with gradient echo and SPECT with 99mTc-ethyl cysteinate dimer using split-dose and subtraction method. In f-MRI study, the significant activation of the sensorimotor area was observed in all subjects. Various concurrent activation regions such as supplementary motor (2 volunteers vs. 4 patients), premotor (1 vs. 2) and ipsilateral sensorimotor area (1 vs. 2) were also observed. In the cases with the activation of neighboring regions of sensorimotor area, it was difficult to identify the area in f-MRI. In SPECT study, the sensorimotor area was depicted as the most increased area in regional cerebral blood flow in eight cases, the characteristics of which were helpful to diagnose the area, while significant activation of the area with edema could not be detected in two patients. Consequently, the sensorimotor area was cross-validated with both the modalities. This study demonstrated that it would be valuable to use both techniques for the clinical assessment of the sensorimotor area.

[Anesthetic management of patients undergoing irrigation and drainage of chronic subdural hematoma--retrospective analysis of analgesia and sedation under locoregional anesthesia].

We retrospectively investigated the relationship between intraoperative hemodynamic variability and variables including patient background, anesthetic profile, and operative profile in 108 patients undergoing irrigation and drainage of chronic subdural hematoma under locoregional anesthesia. Patients were divided into two groups according to the degree of changes in mean arterial blood pressure (MAP) and heart rate (HR) during operation. Group A (n = 66) had MAP and HR changes < 20%, and Group B (n = 42) had MAP or HR changes > or = 20% of preoperative baseline values. Age was significantly higher in group B than Group A (mean +/- SD; 70 +/- 12 vs. 62 +/- 14 years, P < 0.01). In respect to additional anesthetic agents used (none; no additional anesthetic agents, enough; pentazocine > or = 0.3 mg.kg-1 and droperidol > or = 0.05 mg.kg-1, pentazocine > or = 0.5 mg.kg-1 or droperidol > or = 0.15 mg.kg-1, little; less than "enough"), ratio of "little" administered in Group B was significantly higher than that in Group A (64.3% vs. 37.9%, P < 0.05). These findings suggest that intraoperative hemodynamic variability under locoregional anesthesia in patients with chronic subdural hematoma is associated with age and insufficient use of hypnotic and/or analgesic agents.

Three-month oral repeated administration toxicity study of seed saponins of Thea sinensis L. (Ryokucha saponin) in rats.

Ryokucha saponin (RSP), which is one of the ingredients of green tea, was administered orally to rats for 3 months at dose levels of 50, 150 and 500 mg/kg/day to assess any toxic effects. As positive control, 1200 mg/kg/day of quillaia saponin (QSP), which contains saponins equivalent in amount to those in 500 mg RSP/kg/day, was administered. The no-effect level of RSP was 50 mg/kg/day and the lowest-observed-effect level of RSP was 150 mg/kg/day for both sexes, so that the true no-effect level of RSP was estimated to be between these dose levels. In addition, it was confirmed that the toxicity of RSP at 500 mg/kg/day was less than that of QSP at 1200 mg/kg/day and RSP was demonstrated to be safer than QSP, which is a permitted food additive.

A reticuloendothelial system-activating arabinoxylan from the bark of Cinnamomum cassia.

A neutral polysaccharide, named cinnaman AX, was isolated from the dried bark of Cinnamomum cassia Blume. It was homogeneous on electrophoresis and gel chromatography. It is composed of L-arabinose: D-xylose in the molar ratio of 4:3, and its molecular weight was estimated to be about 1.0 x 10(6). Methylation analysis, carbon-13 nuclear magnetic resonance and controlled Smith degradation studies enabled elucidation of its structural features. It showed remarkable reticuloendothelial system-potentiating activity in a carbon clearance test.

Growth, morphology, function, and morphogenetic properties of rat renal glomerular epithelial cells in vitro: effects of retinyl acetate.

We report here that retinyl acetate (RA) regulates growth, morphology, function and cell organization in rat renal glomerular epithelial cells (SGE1). SGE1 cells are able to grow in a serum-free medium (DHFs medium) which is supplemented with insulin, transferrin, selenium, bovine serum albumin (BSA), linoleic acid and epidermal growth factor (EGF). When 0.1 and 1 micrograms/ml RA were added to the medium, the growth rates in the sparse culture were noticeably increased, compared to those in DHFs alone, whereas more than 10 micrograms/ml RA was cytotoxic to the cells. In the confluent culture, addition of 0.1, 1.0 and 10 micrograms/ml RA prolonged the cell survival. Since 10 micrograms/ml RA is not cytotoxic to the confluent culture, the cytotoxic action of RA seems to be dependent on cell density as well as RA dose. Ultrastructural observation revealed that RA treatment caused an increase of microvilli and alteration of cell shape, from flattened to columnar. Biochemical and immunological studies revealed that RA treatment increased the activity of r-glutamyl transpeptidase (GGT) and an amount of the membrane component with molecular mass (Mr) of 108,000 which is identical to one of nephritogenic antigens, Fx1A. By using fluorescence phalloidin stain, it was found that RA treatment increased content and organization of F-actin fibers. Furthermore, in collagen-embedding culture, RA induced 3-dimensional (3D) growth of SGE1 cells leading to the formation of organoids, cystic spheres with central lumen, in a serum-free condition; the addition of DHFs to collagen gel alone was ineffective for the 3D growth.(ABSTRACT TRUNCATED AT 250 WORDS)

A trial of adjuvant combination chemoimmunotherapy for stage III carcinoma of stomach.

A chemoimmunotherapy program designed on the basis of experimental results was administered to 27 patients with stage III carcinoma of stomach following curative resection. The treatment regimen consisted of active immunotherapy with Vibrio cholerae neuraminidase (VCN)-treated autologous tumor cells admixed with bacillus Calmette-Guérin (BCG) and chemotherapy with drugs such as cyclophosphamide (CY), mitomycin C (MMC), and 5-fluorouracil (FU) which proved to enhance the immune response when administered at optimal dose and timing. Then, it was followed by long-term administration of tegafur (FT) and immunomodulators. This treatment significantly improved survival when compared to that of 41 historical control patients treated with surgery alone (P less than 0.001). As compared to 31 control patients concurrently treated with a bolus dose of MMC followed by long-term FT and immunomodulators, survival had a tendency, but not significantly, to be improved in patients treated with this therapy (P less than 0.1). However, the survival rate at 4.5 years was significantly higher than that of control patients (P less than 0.01). These results appeared to show that this type of adjuvant combination chemoimmunotherapy may be of benefit for this group of patients with gastric carcinoma.

Combination chemoimmunotherapy for advanced gastric carcinoma.

Eighty-nine patients with advanced gastric carcinoma were treated with a combination chemo-immunotherapy regimen that consisted of active immunotherapy with Vibrio cholerae neuraminidase (VCN) treated autologous tumor cells admixed with BCG and drugs including cyclophosphamide, mitomycin C (MMC) and 5-fluorouracil, followed by long term tegafur (FT) and immunomodulators. This treatment significantly improved survival rate of patients in Stages III, IV and unresectable or recurrent carcinoma, compared to that of historical controls. As compared to controls treated with MMC followed by long term FT and immunomodulators concurrently, survival rate of those in Stage III tended to improve (P less than 0.1) and survival rate at 4.5 years in Stage III was significantly higher (p less than 0.01), although it was not improved in Stage IV. In patients with unresectable or recurrent tumor, survival time was not significantly lengthened with this therapy when compared with that in patients given BCG alone in the same treatment schedule (CCI-BCG group). However, none of 19 patients in CCI-BCG group survived more than 15 months, although 4 of 28 patients receiving this therapy survived. These results suggest that this combination chemo-immunotherapy is effective for a selected group of patients with advanced gastric carcinoma.

[Studies on As2O3-induced rabbit hypothermia and brain monoamines (author's transl)].

It has been suggested that hypothermia induced in rabbits by As2O3 3 mg/kg (i.v.) depends mostly on the blocking of the thermo-regulatory center. The relationship between hypothermia induced by As2O3 and brain monoamine levels in rabbits was investigated. To clarify the mechanism of the hypothermia, the influence of pretreatment with several agents on As2O3-induced hypothermia and on monoamine levels in the hypothalamus was examined. The core temperature was measured by inserting the thermister probe into the rectum and noradrenaline(NA), 5-hydroxytryptamine(5-HT) and 5-hydroxyindoleacetic acid(5-HIAA) levels in the hypothalamus were estimated fluorometrically. Pretreatment with p-chlorophenylalanine(PCPA), alpha-methyl-p-tyrosine(alpha-MPT) or 5-hydroxytryptophan(5-HTP) did not inhibit the hypothermia induced by As2O3 but did decrease NA levels in the hypothalamus. On the contrary, pretreatment with barbital sodium, pheniprazine, 1-DOPA and 1-tyrosine significantly inhibited the hypothermia or exhibited the hyperthermia. As2O3-induced hypothermia in rabbits was followed by a decrease in NA levels and an increase in 5-HT levels in the hypothalamus. On the other hand, when the hypothermia induced by As2O3 was inhibited by pretreatment with barbital sodium, pheniprazine, 1-DOPA and 1-tyrosine, both NA and 5-HT levels in the hypothalamus were significantly increased. These results suggest that As2O3-induced hypothermia is due to a decrease in NA levels and inhibition of the hypothermia is due to an increase in NA levels, in the rabbit hypothalamus.