Air test as a simple method of screening for Hirschsprung's disease.

AIM: To present the technique and the diagnostic accuracy of the air test to diagnose Hirschsprung's disease (HD). MATERIALS AND METHODS: Children who attended hospital for chronic constipation (CC) between January 2012 and December 2016 for whom the air test was performed were enrolled. The test was conducted during contrast enema under fluoroscopic observation using 20-50 ml injections of air into the rectum through a 10 F Nelaton catheter. The demographics, results of the air test, and additional examinations, as well as the outcomes of subsequent treatments were analysed retrospectively. RESULTS: The air test was conducted in 179 patients (median: 3 years, range: 0-14 years), and was positive in 150 and negative in 29 cases. Of the 29 patients with negative results, four were diagnosed with HD by rectal suction biopsy (RSB). Of the remaining 25 patients, RSB was conducted in seven and HD was excluded in all cases. In all 150 patients with positive air test results, CC was adequately controlled with conservative treatment. The sensitivity and specificity of the air test were 100% (4/4) and 85.7% (150/175), respectively. CONCLUSIONS: The air test can be used as a new non-invasive screening method for HD, performed simultaneously with contrast enema.

Potential bioavailability of phosphorus in citrus orchard soil to Microcystis aeruginosa.

Algal blooms in eutrophic water bodies are controlled by inputs of phosphorus (P) as the growth-limiting nutrient. Runoff particulate P associated with soil from fields often predominates among P fractions. Here, an algal bioassay to investigate the potential bioavailability of particulate P in soil collected from a citrus orchard was conducted. Microcystis aeruginosa was cultured in medium containing soil as the sole source of P. The P in the soil was not notably solubilized after autoclaving. Analyses of chlorophyll-a, suspended solids, particulate organic carbon, and particulate organic nitrogen showed that M. aeruginosa could utilize some of the P present in the soil, perhaps that in particulate form, but this form of P was not sufficient to maintain optimum growth.

The effects of danaparoid, dalteparin and heparin on tissue factor-induced experimental disseminated intravascular coagulation and bleeding time in the rat.

Danaparoid and heparin, on the basis of anti-activated factor X (anti-FXa) activity, were equipotent in accelerating the rate of interaction of FXa and antithrombin III. In rat tissue factor-induced disseminated intravascular coagulation (DIC) models, an intravenous administration of danaparoid inhibited the decrease in plasma fibrinogen and platelet counts and the increase in serum fibrinogen degradation products. Expressed on the basis of anti-FXa activity, these effects were comparable with those of dalteparin and heparin. In rat mesenteric small artery and vein, less bleeding was observed after intravenous administration of danaparoid than after dalteparin or heparin. Danaparoid did not affect adenosine diphosphate- or collagen-induced platelet aggregation, and showed weaker inhibitory effects on aggregation induced by thrombin, or collagen + thrombin, than did dalteparin or heparin. These findings suggest that danaparoid may be useful for the prevention of DIC and has less tendency to cause bleeding than dalteparin or heparin, probably as a result of its weaker ability to inhibit platelet aggregation.

Effects of antitumor activity and protection of shock symptoms by a traditional Chinese medicine (sho-saiko-to) in recombinant human tumor necrosis factor administered mice.

The effects of a traditional Chinese medicine Sho-saiko-to (Kampo prescription) were investigated on the various metabolic disorders and antitumor activity of recombinant human tumor necrosis factor (rhTNF) administered to mice. The glycogen level in liver of rhTNF (5 x 10(4) units/mouse, i.v.)-injected mice was markedly lower at 4 h post-intoxication than that in the control, whereas the administration of rhTNF to Sho-saiko-to (500 mg/kg/d, p.o.)-pretreated mice resulted in a greater level of glycogen than that in rhTNF alone-treated mice. In mice pretreated with Sho-saiko-to, the level of fibrinogen 4 h after rhTNF injection markedly increased as compared to that in mice treated with rhTNF alone. We also estimated the NO2 in murine macrophage cell line J774A.1 using mice serum after administration of Sho-saiko-to. Our results clearly demonstrated that J774A.1 cells stimulated with endotoxin (1 micrograms/ml) and rhTNF (1 x 10(4) units/ml) can effectively produce nitric oxide (NO), and ascertained the suppressive effect of Sho-saiko-to (500 mg/kg/d, p.o)-pretreated serum on NO generation by endotoxin/TNF-activated J774A.1 cells. When the cells were incubated with endotoxin/TNF and Sho-saiko-to pretreated serum (10-100 microliters), the NO level was significantly lower than that in control serum incubated with endotoxin/TNF alone. The effect of Sho-saiko-to (1 and 10 micrograms/ml) on in vitro cytotoxicity by rhTNF in Meth-A Sarcoma cells was observed to be in a dose dependent fashion. In addition, there was a remarkable enhancement of antitumor activity of rhTNF by Sho-saiko-to pretreatment in mice. These findings suggest that the Kampo prescription Sho-saiko-to may protect mice from severe shock syndrome by rhTNF, and that it may enhance rhTNF-induced activity.

Indonesian medicinal plants. XVII. Characterization of quassinoids from the stems of Quassia indica.

Four new quassinoids named samaderines X (1), Y (2) and Z (3), and indaquassin X (5), and a new C19 quassinoid glycoside, 2-O-glucosylsamaderine C (10), together with five known quassinoids, samaderines B (7), C (8), and E (4), indaquassin C (6), and simarinolide (9), were isolated form the stems of Quassia indica (Simaroubaceae), an Indonesian medicinal plant. The chemical structures of these quassinoids have been elucidated on the bases of their chemical and physiochemical properties. Samaderines X (1), Z (3), E (4), and B (7) were shown to exhibit significant growth-inhibitory activity against the cultured malarial parasite Plasmodium falciparum (a chloroquine- resistant K1 strain), and 1--8 were shown to exhibit in vitro cytotoxicity (IC50: 0.04--100 micrograms/ml) against KB cells. Samaderines X (1), B (7) and C (8), as well s indaquassin X (5), exhibited inhibitory activity in the in vitro endothelial cell-neutrophil leukocyte adhesion assay, whereas samaderines X (1) and B (7) were found to exhibit significant anti-inflammatory activity.

Modification of cultured Madin-Darby canine kidney cells with dietary unsaturated fatty acids and regulation of arachidonate cascade reaction.

Madin-Darby canine kidney (MDCK) cells were modified with dietary unsaturated fatty acids. The effects on the fatty acid composition in each phospholipid class and the formation of prostanoids upon stimulation were studied, from which the specificity of metabolism of individual unsaturated fatty acids and the regulation of arachidonate cascades in the modified cells were discussed. C18 unsaturated fatty acids were preferentially incorporated into phosphatidylcholine (PC) over phosphatidylethanolamine (PE), but arachidonic acid (20:4(n-6)) derived from gamma-linolenic acid (18:3(n-6)) was much more predominant in PE than PC. The fatty acid level in PE ranged from about 26-28% when the cells were modified with 20:4(n-6) or 5,8,11,14,17-eicosapentaenoic acid (20:5(n-3)), indicating the limitation of the storage of the eicosapolyenoic acids. The extra amounts appeared to be stored in PC. 18:3(n-6) was comparable to 20:4(n-6) to raise the level of 20:4(n-6) in PE, but not in PC which had half of 20:4(n-6) in PE. The supplementation of linoleic acid (18:2(n-6)). 18:3(n-6), and 20:4(n-6) caused significant increases in the synthesis of prostaglandin (PG)E2 up to almost the same levels when the modified cells were stimulated with 50 nM PMA and 100 nM A23187 for 24h. The cultured cells modified with eicosapolyenoic acids including 20:3(n-6), 20:4(n-6), and 20:5(n-3) were found to be inhibitory for the induction of PGE2 synthetic activity involving de nova synthesis of PG endoperoxide synthase, suggesting negative feedback regulation of the modified cells.

Satisfactory remission achieved by PUVA therapy in a case of crisis-type adult T-cell leukaemia/lymphoma with generalized cutaneous leukaemic cell infiltration.

We used PUVA therapy in a patient with crisis-type adult T-cell leukaemia/lymphoma and generalized cutaneous leukaemic cell infiltration. PUVA proved very effective in reducing leukaemic cells and in clearing the eruption. To understand the way in which PUVA produced a reduction in the number of leukaemic cells, we examined peripheral blood cells by light and electron microscopy. Light microscopy was of little help, but electron microscopy revealed that PUVA induced apoptosis-like changes in circulating leukaemic cells. This suggests that apoptosis-like changes in leukaemic cells might be the reason for the success of this treatment.

Depressive effect of a traditional Chinese medicine (sho-saiko-to) on endotoxin-induced nitric oxide formation in activated murine macrophage J774A.1 cells.

The present study investigated whether or not Sho-saiko-to (crude powder extract, TJ-9) can suppress nitric oxide (NO) generation by endotoxin-activated J774A.1 cells in order to study the preventive mechanism of Sho-saiko-to against endotoxemia. In this experiment, we estimated the NO2- in the murine macrophage cell line J774A.1 using the Griess method. Our results clearly demonstrated that J774A.1 cells stimulated with endotoxin (0.01-10 micrograms/ml) can effectively produce NO, and the production was dependent on the dose of endotoxin. On the other hand, we investigated the suppressive effect of TJ-9 (10-100 micrograms/ml) on NO generation by endotoxin (0.1 microgram/ml)-activated J774A.1 cells. The NO level when the cells were incubated with endotoxin and TJ-9 (10-20 micrograms/ml) was slightly lower than that in cells treated with endotoxin alone. In contrast, treatment with TJ-9 (50-100 micrograms/ml) significantly inhibited endotoxin-activated NO generation in J774A.1 cells, whereas the treatment with TJ-9 (10-100 micrograms/ml) alone was ineffective in inducing NO formation and in inhibiting cell viability in the J774A.1 cells. These findings suggest that a Kampo presciption of Sho-saiko-to shows a suppressive effect on NO generation in macrophages stimulated with endotoxin, and that it may be useful in improving endotoxin-shock symptoms.

The effects of KBT-3022, a new anti-platelet agent, on hemorheological properties in guinea pigs.

Using guinea pigs, a study was conducted on the effects of KBT-3022, a new anti-platelet agent, on hemorheological properties in various tests including blood filterability, blood viscosity, shear stress-induced red blood cell (RBC) deformability and contents of ATP and 2,3-diphosphoglycerate (2,3-DPG). Oral administration of KBT-3022 at 1 and 10 mg/kg significantly increased blood filterability, and significantly reduced blood viscosity at 10 mg/kg without changing the hematocrit, plasma fibrinogen concentration or plasma viscosity. KBT-3022 (10 mg/kg, p.o.) improved RBC deformability in response to shear stress, which was evoked by passing the blood through a thin tube. This dose of KBT-3022 also increased the contents of ATP and 2,3-DPG in RBC. These findings indicate that KBT-3022 may reduce blood viscosity as a sequel to improvement of RBC deformability through direct action on RBC. The increase in the intracellular levels of ATP and 2,3-DPG was considered to be involved in this improvement of hemorheological properties. These hemorheological effects of KBT-3022 appear to be promising for the treatment of patients with ischemic vascular disease.

Defense effects of a traditional Chinese medicine (sho-saiko-to) against metabolic disorders during endotoxemia; approached from the behavior of the calcium ion.

The present study was carried out as an approach from intracellular Ca2+ to clarify the preventive effects of a traditional Chinese medicine, Sho-saiko-to (Kampo prescription, TJ-9), against various metabolic disorders during endotoxemia. In this experiment, we estimated the cytosolic-free Ca2+ concentration ([Ca2+]i) in liver single cells using a photonic microscope system. The [Ca2+]i in a single liver cell of endotoxin (6 mg/kg, i.p.)-injected mice was greater at 18 h post-intoxication than that in the control, whereas the administration of endotoxinin to TJ-9 (500 mg/kg/d, p.o.)-pretreated mice resulted in a markedly lower level of [Ca2+]i than that in endotoxin-treated mice. In the mice pretreated with TJ-9, the CA(2+)-ATPase activity in liver plasma membrane 18 h after endotoxin injection was markedly increased as compared to that in the endotoxin-treated mice. By contrast, the Ca(2+)-ATPase activity in liver mitochondria was lower in endotoxemic mice pretreated with TJ-9 than in mice given endotoxin alone. State 3 respiration and the respiratory control index (RCI), which are the parameters of mitochondrial function, were 41% and 35% lower, respectively, in the liver of mice given endotoxin than those levels in the control. However, the levels of state 3 and RCI in endotoxin-TJ-9-treated mice were markedly increased as compared to those of the endotoxin-treated mice. These findings suggest the protective effect of TJ-9 in the damage of liver mitochondrial function in endotoxin-poisoned mice.(ABSTRACT TRUNCATED AT 250 WORDS)

Preventive effects of a traditional Chinese medicine (sho-saiko-to) against oxygen toxicity and membrane damage during endotoxemia.

The preventive effects of a traditional Chinese medicine Sho-saiko-to (Kampo prescription, TJ-9) were determined from oxygen toxicity and membrane damage in liver during endotoxemia. The liver lipid peroxide level and xanthine oxidase activity 18 h after administration of endotoxin (6 mg/kg, i.p.) to TJ-9 (500 mg/kg/d, p.o.)-pretreated mice were markedly lower than that in endotoxin-treated mice, whereas the administration of TJ-9 significantly increased superoxide dismutase and glutathione peroxide activities in liver of endotoxin-injected mice. In the mice pretreated with a TJ-9, the levels of alpha-tocopherol and nonprotein SH in liver tissue 18 h after endotoxin injection were markedly increased as compared to those in endotoxin-treated mice. Leakages of acid phosphatase and lactate dehydrogenase isozyme in serum were markedly lower in endotoxin-TJ-9-treated mice than those in mice given endotoxin. The administration of TJ-9 clearly prevented the membrane protein damage arising from endotoxin challenge. Kampo prescription Sho-saiko-to thus appears to protect the liver plasma membrane from injury by free radicals which occur in a tissue ischemic state during endotoxemia.

Preventive effects of a Chinese herb medicine (sho-saiko-to) against lethality after recombinant human tumor necrosis factor administration in mice.

We observed the effects of a chinese herb medicine Sho-saiko-to on the lethal and antitumor activities of recombinant human tumor necrosis factor (rhTNF) administered in mice. Sho-saiko-to was noted to protect the rhTNF-induced lethality in galactosamine-hypersensitized mice, and also Sho-saiko-to pretreated mice was protected against the decrease of rectal temperature after rhTNF administration. On the other hand, there was a remarkable enhancement of antitumor activity of rhTNF by Sho-saiko-to pretreatment. These results suggest that Sho-saiko-to drug may protect mice from severe shock syndrome induced by rhTNF.

The amino acid sequences of proteinase inhibitors I-A and I-A' from adzuki beans.

Several proteins which strongly inhibit trypsin have been found in adzuki bean seeds. Two of them, designated as adzuki proteinase inhibitors (API) I-A and I-A', were analyzed for their amino acid sequences by conventional methods. Inhibitors I-A and I-A' exhibited strong homology with other Bowman-Birk type proteinase inhibitors from leguminous seeds in spite of belonging to different genera. Inhibitors I-A and I-A' consisted of 78 and 72 amino acid residues and their molecular weights were 9,100 and 8,300, respectively. Inhibitor I-A' lacked the six amino acid residues of the amino terminus of inhibitor I-A and had an asparagine residue in place of the aspartic acid residue at position 40 of inhibitor I-A. The results showed the occurrence of some genetic variants of proteinase inhibitors in adzuki bean seeds. Inhibitor I-A was a double-headed one, and the reactive sites for trypsin were Lys-Ser and Arg-Ser bonds. Therefore, inhibitor I-A' was also assumed to be a double-headed one having Lys-Ser and Arg-Ser bonds as the reactive sites for the enzyme.