[Extended thymectomy and intraoperative-intrapleural perfusion hyperthermo-chemotherapy for stage IVa invasive thymoma with myasthenia gravis].

A 37-year-old woman diagnosed with ocular myasthenia gravis was referred to our department. Chest computed tomography (CT) showed anterior mediastinal tumor and right pleural dissemination. Extended thymectomy and right intraoperative-intrapleural perfusion hyperthermo-chemothrapy (IPHC) were performed. Pathological diagnosis was invasive thymoma type B2 and stage IVa based on Masaoka's classification. The post operative course was uneventful. The patient underwent 4 cycles of adjuvant chemotherapy with doxorubicin, cisplatin, vincristine, and cyclophosphamide (ADOC), and is free from recurrence at 12 months postoperatively.

Shosaikoto increases calprotectin expression in human oral epithelial cells.

BACKGROUND AND OBJECTIVE: Oral epithelial cells help to prevent against bacterial infection in the oral cavity by producing antimicrobial peptides (AMPs). A broad-spectrum AMP, calprotectin (a complex of S100A8 and S100A9 proteins), is expressed by oral epithelial cells and is up-regulated by interleukin-1alpha (IL-1alpha). Shosaikoto (SST) is a traditional Japanese herbal medicine that has immunomodulatory effects and is reported to enhance the levels of IL-1alpha in epithelial cells. The purpose of this study was to investigate the effect of SST on the expression of calprotectin and other AMPs through the regulation of IL-1alpha in oral epithelial cells. MATERIAL AND METHODS: Human oral epithelial cells (TR146) were cultured with SST (at concentrations ranging from 10 to 250 microg/mL) in the presence or absence of anti-IL-1alpha or IL-1 receptor antagonist. The expression of S100A8- and S100A9-specific mRNAs was examined by northern blotting. Calprotectin expression and IL-1alpha secretion were investigated by immunofluorescent staining or ELISA. The expression of other AMPs and IL-1alpha was analyzed by RT-PCR and by quantitative real-time PCR. RESULTS: Shosaikoto (25 microg/mL) significantly increased the expression of S100A8- and S100A9-specific mRNAs and calprotectin protein. Shosaikoto increased S100A7 expression, but had no effect on the expression of other AMPs. The expression of IL-1alpha-specific mRNA and its protein were slightly increased by SST. A neutralizing antibody against IL-1alpha or IL-1 receptor antagonist inhibited SST up-regulated S100A8/S100A9 mRNA expression. CONCLUSION: These results suggest that SST increases the expression of calprotectin and S100A7 in oral epithelial cells. In response to SST, up-regulation of calprotectin may be partially induced via IL-1alpha.

Regulation of calprotectin expression by interleukin-1alpha and transforming growth factor-beta in human gingival keratinocytes.

BACKGROUND AND OBJECTIVE: Calprotectin, a heterodimer of S100A8 and S100A9 with antimicrobial properties, is expressed in gingival keratinocytes and plays an important role in innate immunity. Because calprotectin expression is localized in the spinous cell layer of the gingival epithelium, we hypothesized that the expression of calprotectin in keratinocytes is related to the differentiation stage. The aim of the present study was to investigate the relationship between calprotectin expression and keratinocyte differentiation using some factors that regulated its differentiation. MATERIAL AND METHODS: Normal human gingival keratinocytes were isolated from gingival tissues obtained at the extraction of wisdom teeth, and were cultured in serum-free keratinocyte medium supplemented with interleukin-1alpha or calcium, which promote keratinocyte differentiation, and transforming frowth factor-beta (TGF-beta) or retinoic acid, which suppress its differentiation. The expression of S100A8/A9 mRNA and the production of calprotectin in normal human gingival keratinocytes were examined by northern blotting and enzyme-linked immunosorbent assay, respectively. The expression of cytokeratin 14, involucrin and filaggrin (marker proteins of keratinocyte differentiation) was investigated by immunohistochemical staining, and the DNA-binding activity of CCAAT/enhancer binding protein alpha (C/EBPalpha), a transcription factor, was examined by electrophoretic mobility shift assay. RESULTS: The expression of S100A8/A9 mRNA and the production of calprotectin were increased by interleukin-1alpha and calcium, but decreased by TGF-beta. RA inhibited the expression of S100A8/A9 and keratinocyte differentiation, which were induced by interleukin-1alpha. C/EBPalpha DNA-binding activity in normal human gingival keratinocytes was enhanced by interleukin-1alpha and calcium, but suppressed by TGF-beta. CONCLUSION: The present study suggests that calprotectin expression is related to keratinocyte differentiation and that C/EBPalpha is a regulator of calprotectin expression in keratinocytes.

Cytokine and antibody production during the course of resolution in Plasmodium yoelii 17XL-infected BALB/c mice treated with febrifugine and isofebrifugine mixture from leaves of Hydrangea macrophylla var. Otaksa.

Cytokine and antibody production was investigated during the course of resolution of primary infection in Plasmodium yoelii 17XL-infected BALB/c mice treated with a mixture of febrifugine and isofebrifugine. The infected mice in an untreated control group showed a progressively increasing parasitemia, leading to mouse death. In contrast, infected mice given the mixture orally showed low parasitemia levels during administration. Following a transient increase in parasitemia in the bloodstream of the treated mice, no parasites could be detected by microscopic examination. Analysis of cytokines in plasma showed that the plasma IFN-gamma levels elevated significantly within the first week of infection in both groups. Furthermore, on day 20 the plasma IFN-gamma and IL-4 levels elevated significantly in the treated mice and the production of both cytokines was sustained until at least day 40. The production of both cytokines in the treated mice was coincident with a decrease in parasitemia. The production of parasite-specific antibodies in the course of P. yoelii 17XL infection was also monitored. In the drug-treated mice, the titers of parasite-specific IgG1, IgG2a, IgG2b and IgG3 elevated significantly from day 20; and the production of parasite-specific antibodies was coincident with a decrease in parasite numbers in the bloodstream.

Overexpression of pyrimidine nucleoside phosphorylase enhances the sensitivity to 5'-deoxy-5-fluorouridine in tumour cells in vitro and in vivo.

5-Fluorouracil (5-FU) and 5'-deoxy-5-fluorouridine (5'-DFUR), a prodrug of 5-FU, are representative of the chemotherapeutic agents for colorectal adenocarcinomas. Pyrimidine nucleoside phosphorylase (PyNPase) catalyses the conversion of 5'-DFUR to 5-FU, the activated form. Murine adenocarcinoma CT26 cells were transfected with human PyNPase cDNA. The engineered transfectants producing PyNPase augmented the response to 5'-DFUR in vitro and in vivo. Animals were administered by means of intraperitoneal (i.p.) injection, and not orally, in order to obtain a better efficiency of absorption. The tumours of the transfected cells nearly all disappeared, even following treatment with quite a small amount of the anticancer agent. The animals injected with the tranfected cells were protected against subsequent challenge with the parental tumour cell line. These findings demonstrate that PyNPase gene transfection increases the sensitivity to 5'-DFUR, and thereby decreases the toxicity of the agent.

Prophylactic chemolipiodolization for postoperative hepatoma patients.

BACKGROUND/AIMS: The efficacy of prophylactic chemolipiodolization following hepatic resection in patients with hepatocellular carcinoma was studied. METHODOLOGY: Forty-four of 67 consecutive patients with hepatocellular carcinoma who underwent hepatectomy between 1980 and 1997 were divided into two groups: group A (n = 21), in which prophylactic chemolipiodolization was performed during postoperative follow-up (2.4 times on average using a 39 mg mean dose of epirubicin or doxorubicin); and group B (n = 23), without prophylactic chemolipiodolization. The clinicopathological background and patient survival were compared retrospectively. RESULTS: There were no differences in the clinicopathological background between the two groups. Multiple intrahepatic recurrence was frequently observed in group B (P < 0.02). The recurrence-free survival rates in group A (54.4% and 31.1% at 3 and 5 years, respectively) were significantly higher than those in group B (15.7% and 7.9%, respectively). The survival rates of group A (95.2% and 80.4% at 3 and 5 years, respectively) were significantly higher than those in group B (40.1% and 22.9%, respectively). CONCLUSIONS: Our data suggest that postoperative prophylactic chemolipiodolization can be an effective treatment in reducing intrahepatic recurrence and may prolong survival for hepatocellular carcinoma patients following hepatic resection.

Moxibustion treatment of breech presentation.

Breech presentation was successfully corrected by stimulating acupuncture points with moxibustion or low-frequency electrical current. Only patients with breech pregnancies at the 28th week or later were entered into the study. With moxibustion treatment, the control group had a spontaneous correction rate of 165/224 (73.66%), and the treatment group had a correction rate of 123/133 (92.48%) (P<0.0001, x2 test). With low-frequency percutaneous electrical stimulation, the correction rate was 20/941 (83.87%) in the control group and 171/191 (89.52%) in the treatment group (P=0.094, x2 test). The controls in the moxibustion study did no exercises and received no external manipulation to correct breech presentation whereas those in the electrical stimulation study experienced both. Acupuncture stimulation, especially with moxibustion, is expected to serve as a safe and effective modality in the management of breech presentation in a clinical setting.

Ghrelin is an appetite-stimulatory signal from stomach with structural resemblance to motilin.

BACKGROUND & AIMS: : Ghrelin, an endogenous ligand for growth hormone secretagogue receptor, was recently identified in the rat stomach. We examined the effects of the gastric peptide ghrelin on energy balance in association with leptin and vagal nerve activity. METHODS: : Food intake, oxygen consumption, gastric emptying, and hypothalamic neuropeptide Y (NPY) messenger RNA expression were measured after intra-third cerebroventricular or intraperitoneal injections of ghrelin in mice. The gastric vagal nerve activity was recorded after intravenous administration in rats. Gastric ghrelin gene expression was assessed by Northern blot analysis. Repeated coadministration of ghrelin and interleukin (IL)-1 beta was continued for 5 days. RESULTS: : Ghrelin exhibited gastroprokinetic activity with structural resemblance to motilin and potent orexigenic activity through action on the hypothalamic neuropeptide Y (NPY) and Y(1) receptor, which was lost after vagotomy. Ghrelin decreased gastric vagal afferent discharge in contrast to other anorexigenic peptides that increased the activity. Ghrelin gene expression in the stomach was increased by fasting and in ob/ob mice, and was decreased by administration of leptin and IL-1 beta. Peripherally administered ghrelin blocked IL-1 beta-induced anorexia and produced positive energy balance by promoting food intake and decreasing energy expenditure. CONCLUSIONS: : Ghrelin, which is negatively regulated by leptin and IL-1 beta, is secreted by the stomach and increases arcuate NPY expression, which in turn acts through Y(1) receptors to increase food intake and decrease energy expenditure. Gastric peptide ghrelin may thus function as part of the orexigenic pathway downstream from leptin and is a potential therapeutic target not only for obesity but also for anorexia and cachexia.

Dietary intake of rapeseed oil or soybean oil as the only fat nutrient in spontaneously hypertensive rats and Wistar Kyoto rats - blood pressure and pathophysiology.

Spontaneously hypertensive rats (SHR) and Wistar Kyoto (WKY) rats were fed a diet containing 10% rapeseed (canola) oil or soybean oil as dietary fat, and given drinking water containing 1% NaCl for 26 weeks. From the 10th week and later, systolic blood pressure in the canola oil group became higher than that in the soybean oil group in each strain. The 26-week feeding of canola oil increased plasma lipids and the neutrophil counts, and decreased the platelet counts. In the canola oil group the heart and kidney tended to become heavier with sporadically found histologic lesions. Acetylcholine- and nitroprusside-induced dilating responses of isolated aortic rings and norepinephrine- and veratridine-induced increases in vascular tone of isolated perfused mesenteric arteries were not different between the two groups in each strain. These results demonstrate that canola oil intake as the only dietary fat elevates blood pressure of the rat provided with drinking water containing 1% NaCl through mechanisms other than blunt dilating response of the blood vessel due to dysfunction of the endothelium or vascular smooth muscle, the augmented response to norepinephrine in the arteries and the increased amount of norepinephrine in the sympathetic nerve endings. The lesions in the heart and kidney in SHR may be related to a strain-specific peripheral vascular deterioration which was disclosed by the extremely high blood pressure in the canola oil group.

A lectin from an edible mushroom Pleurotus ostreatus as a food intake-suppressing substance.

In an experiment in which rats were allowed free access to food and water, the rats did not eat the diet containing a mushroom Pleurotus ostreatus even if they were emaciated. A P. ostreatus lectin (POL) was isolated from the mushroom as the food intake-suppression principle. In hemagglutination inhibition assays, Me-alphaGalNAc was the most potent inhibitor among the monosaccharides tested. Among all the sugars tested, 2'-fucosyllactose (Fucalpha1-->2Galbeta1-->4Glc) was the strongest inhibitor and its inhibitory potency was five times greater than that of Me-alphaGalNAc. POL exhibited a binding ability to bovine submaxillary mucin (BSM) and asialo-BSM and the other glycoproteins were inert to the binding. The food intake-suppressing activity of POL was dependent on the dose. The diet containing 0.1% POL caused a 50% decrease in the food intake of rats against the control.

Effects of polysaccharides (FI0-b) from mycelium of Ganoderma tsugae on proinflammatory cytokine production by THP-1 cells and human PBMC (I).

AIM: To compare the effects of water-soluble polysaccharides, FI0-b, and its formic acid-modified derivative, FI0-b-H, on production of human proinflammatory cytokines. METHODS: The polysaccharides were modified by formic acid. Cytokine production was quantitated by radioimmunoassay. mRNA for cytokines was measured by semi-quantitative RT-PCR. RESULTS: FI0-b and FI0-b-H 4, 40, and 400 mg/L significantly downregulated interleukin-1 alpha (IL-1 alpha) production by THP-1 cells induced by lypopolysaccharide (LPS) 1 or 10 mg/L and phorbol myristate acetate (PMA) 200 nmol/L. At lower stimulation with LPS 10 mg/L and PMA 200 nmol/L, both polysaccharides significantly upregulated tumor necrosis factor alpha (TNF alpha) production by THP-1 cells. However, at higher stimulation with LPS 100 mg/L and PMA 200 nmol/L, they downregulated TNF alpha production. FI0-b-H downregulated interleukin-8 (IL-8) production by THP-1 cells at a lower-dose of LPS 1 mg/L and PMA 200 nmol/L, but upregulated IL-8 production at a higher-dose of LPS 10 mg/L and PMA 200 nmol/L. Production of cytokines (IL-1 alpha and TNF alpha) was transcriptionally or post-transcriptionally regulated by FI0-b and FI0-b-H. CONCLUSION: The water-soluble polysaccharides of Ganoderma tsugae mycelium have bidirectional immunomodulatory effects on cytokine production in different stimulatory conditions in a dose-dependent manner. Compared with FI0-b, FI0-b-H has more marked effects on human proinflammatory cytokine production.

Effects of polysaccharides (FI0-c) from mycelium of Ganoderma tsugae on proinflammatory cytokine production by THP-1 cells and human PBMC (II).

AIM: To study the effects of water-soluble polysaccharides. FI0-c, and its sulfated derivative, FI0-c-S, on production of human proinflammatory cytokines, interleukin-1 alpha (IL-1 alpha) and tumor necrosis factor alpha (TNF alpha). METHODS: The herbal polysaccharides were modified by chlorosulfornic acid in dimethyl sulfoxide (Me2SO). Cytokine production was measured by radioimmunoassay, mRNA for the cytokines was measured by semi-quantitative RT-PCR. RESULTS: FI0-c 4 mg/L itself induced IL-1 alpha production by THP-1 cells without stimulants, such as lipopolysaccharides (LPS) and phorbol myristate acetate (PMA). On the other hand, FI0-c and FI0-c-S inhibited the IL-1 alpha production by THP-1 cells with these stimulants. FI0-c and FI0-c-S significantly upregulated TNF alpha production by THP-1 cells without stimulants or at a low dose of LPS 10 mg/L and PMA 200 nmol/L, whereas these polysaccharides markedly downregulated the TNF alpha production by a high dose of LPS 100 mg/L and PMA. Human peripheral blood mononuclear cells (PBMC) responded to FI0-c and FI0-c-S in IL-1 alpha and TNF alpha production in a fashion similar to THP-1 cell responses. FI0-c 4 mg/L downregulated high-dose LPS- and PMA-induced IL-1 alpha or TNF alpha mRNA and their protein production by THP-1 cells. CONCLUSION: The water-soluble polysaccharides of Ganoderma tsugae mycelium have bidirectional immunomodulatory effects on cytokine production in different cell stimulatory conditions. Chemical modification of this polysaccharide changed the intensity of regulatory effect on cytokine production.

Mumefural, citric acid derivative improving blood fluidity from fruit-juice concentrate of Japanese apricot (Prunus mume Sieb. et Zucc).

The effects of food components on blood fluidity were studied by in vitro assay using a dedicated microchannel instrument for model capillaries. We found that the fruit-juice concentrate of the Japanese apricot (Prunus mume Sieb. et Zucc), a traditional Japanese food, markedly improved the fluidity of human blood. Using HPLC, we isolated the active compounds and characterized them using UV, MS, IR, and NMR. They included a novel compound, 1-[5-(2-formylfuryl)methyl] dihydrogen 2-hydroxypropane-1,2, 3-tricarboxylate (mumefural), and a related compound, 5-hydroxymethyl-2-furfural (HMF). Mumefural markedly improved blood fluidity in all subjects, while HMF worked differently in different individuals. The flow rate of blood spiked with mumefural or HMF was compared to that of the two predominant organic acids in the fruit. Citric acid, malic acid, and furfuryl alcohol also improved fluidity in all subjects. The activity of P. mume is derived from not only artifacts produced during thermal processing, such as mumefural, but also from endogenous organic acids.

Extraction and identification of antioxidants in the roots of yacon (Smallanthus sonchifolius).

Yacon, Smallanthus sonchifolius (Poepp. & Endl.) H. Robinson, Asteraceae, an important economic species grown for its juicy tuberous root, is potentially beneficial in the diet to diabetics. The antioxidative activity of yacon root was studied by 1, 1-diphenyl-2-picrylhydrazyl (DPPH) assay. Antioxidants were extracted by methanol and isolated and purified by gel permeation chromatography and preparative reverse-phase HPLC. Two of the major antioxidants were identified as chlorogenic acid and tryptophan by NMR and mass spectrometry.

Phosphate as a limiting factor for the cell division of tobacco BY-2 cells.

The re-addition of phosphate to tobacco BY-2 cells deprived of phosphate for 3 d induced cells to semi-synchronously re-enter the cell cycle from a static state. Though the addition of auxin to auxin-starved tobacco BY-2 cells also induced cell division (Ishida et al. 1993), some major differences were observed between these two systems. BY-2 cells lost the ability to re-enter the cell cycle after prolonged periods of auxin deprivation, but in contrast retained this ability after longer periods in the absence of phosphate. By differential cDNA screening we identified a phosphate-induced gene phi-1. phi-1 was rapidly induced by the addition of phosphate with transcript levels starting to decrease by the start of DNA synthesis. phi-1 does not share any significant homology with any gene with known functions over its full length. However, the N-terminus shared some homology with plasma membrane ATPases suggesting that it may be involved in some process of phosphorylation. Immunolocalization of the phi-1 gene product revealed that it rapidly accumulated in the cytoplasm prior to the start of plastid and nuclear DNA synthesis. These results are discussed in relation to the role of phosphate in inducing plant cell division.

Antioxidative activities in some common seaweeds.

Dietary antioxidants from plants are believed to help prevent aging and other diseases through radical scavenging activity. Free radical scavenging activities by 1,1-diphenyl-2-picrylhydrazyl (DPPH) assay and deoxyribose assay were ascertained among 27 species of common seaweeds using a sequential extraction method. The results indicated that, DPPH scavenging activity existed in 15 species; Gelidium amansii, Gloiosiphonia capillaris, Polysiphonia urceolata, Sargassum kjellmanianum, Desmarestia viridis, and Rhodomela teres showed the strongest activities. On the other hand, the deoxyribose results showed that almost all the seaweed species had good ability to scavenge hydroxyl radicals. The most active species were Rhodomela teres and Chorda filum.

Inhibition of arachidonate lipoxygenase activities by 2-(3,4-dihydroxyphenyl)ethanol, a phenolic compound from olives.

The effects of olive fruit extract on arachidonic acid lipoxygenase activities were investigated using rat platelets and rat polymorphonuclear leukocytes (PMNL). Olive extract strongly inhibited both 12-lipoxygenase (12-LO) and 5-lipoxygenase (5-LO) activities. One of the compounds responsible for this inhibition was purified and identified as 2-(3,4-dihydroxyphenyl)ethanol (DPE). DPE inhibited platelet 12-LO activity (IC50, 4.2 microM) and PMNL 5-LO activity (IC50, 13 microM) but not cyclooxygenase activity in cell-free conditions. It also inhibited 12-LO activity in intact platelets (IC50, 50 microM) and reduced leukotriene B4 production in intact PMNL stimulated by A23187 (IC50, 26 microM). The inhibition by DPE of both lipoxygenase activities was stronger than that by oleuropein, caffeic acid, or 7 other related phenolic compounds, especially in intact cells. These results suggest that DPE is a potent specific inhibitor of lipoxygenase activities.

Screening of antipsychotic drugs by wide-bore capillary gas chromatography with nitrogen phosphorus detection--detection levels in plasma.

The detection levels of 12 antipsychotic drugs as ascertained from drug screening were examined in order to use the screening results effectively for treatment of patients or medico-legal examinations in cases of poisoning. Wide-bore capillary gas chromatography with nitrogen phosphorus detection was used for the analysis. The detection limits of drugs in plasma using 3 extraction procedures, single-solvent, 3-step solvent and solid-phase extractions were 50-100 ng/ml, 10-200 ng/ml and 5-50 ng/ml, respectively. The detection limits were compared with blood levels previously reported in the literature. Only 4 drugs were detected at the therapeutic level, thioridazine, floropipamide, sultopride and oxypertine using the single-solvent extraction procedure. Using the 3-step solvent extraction procedure, chlorpromazine, levomepromazine and zotepine in addition to the above 4 drugs were detected. Using the solid-phase extraction procedure, 8 of 12 drugs were able to be detected at the therapeutic level, with the exception of perphenazine, haloperidol, bromperidol and nemonapride. These data were proved to be useful for a rapid forensic diagnosis.

Thyroid hormone suppresses the differentiation of osteoprogenitor cells to osteoblasts, but enhances functional activities of mature osteoblasts in cultured rat calvaria cells.

The effects of thyroid hormone on osteoblastic differentiation and activity were studied in fetal rat calvaria (RC) cells cultured for up to 30 days in medium supplemented with thyroid hormone-depleted serum. In this condition, the cells proliferated and differentiated to form mineralized bone nodules (BN) and expressed osteoblastic markers such as alkaline phosphatase (ALP), osteocalcin (OCN), and osteopontin (OPN). The continuous presence of triiodothyronine (T3) at 10(-9)-10(-8) M in the medium inhibited the osteoblastic differentiation: 34% decrease in ALP activity on day 12 and 60% decrease in BN formation on day 15 at 10(-8) M. T3 at these doses had no effect on the DNA content of RC cells at confluence (day 6). Short-term (48-h) exposure of T3 at 10(-9) M or higher decreased ALP activity when RC cells were differentiating (days 7-11). However, when BN formation by the cells had already reached a plateau (day 28), the activity was increased by treatment with T3 at 10(-7)-10(-6) M. OCN production was increased dose dependently by this treatment with T3 (2.1-fold and 1.3-fold of control at 10(-8) M on days 11 and 28, respectively). Similar increases were observed in the levels of OCN mRNA. In addition, increases in phosphorylated OPN in the medium (day 11) and mineralized matrix (day 28) were observed (1.5-fold at 10(-8)-10(-6) M), while OPN synthesis and the level of its mRNA were depressed by T3 (60-70% of control at 10(-8) M). These results suggest that T3 regulates osteoblastic differentiation and activity depending on the state of cell differentiation: T3 suppresses the differentiation of osteoprogenitor cells to osteoblasts, but enhances the functional activity of mature osteoblasts.