Stable progeny production of the amphidiploid resynthesized Brassica napus cv. Hanakkori, a newly bred vegetable.

Resynthesized Brassica napus cv. Hanakkori (AACC, 2n = 38) was produced by cross-hybridization between B. rapa (AA, 2n = 20) and B. oleracea (CC, 2n = 18) as a new vegetative crop. Many studies have provided evidences for the instability and close relationship between A and C genome in the resynthesized B. napus cultivars. In fact, seed produced to obtain progeny in Hanakkori had unstable morphological characters and generated many off-type plants. In this study, we investigated the pollen fertility, chromosome number, structure, and behavior linked to various Hanakkori phenotypes to define factors of unstable phenotypic expression in the progeny. Hanakkori phenotypes were categorized into five types. The results of pollen fertility, chromosome number, and fluorescence in situ hybridization analysis for somatic mitosis cells indicated that the off-type plants had lower pollen fertility, aberrant chromosome number, and structures with small chromosome fragments. Observation of chromosomes at meiosis showed that the meiotic division in off-type plants led to appreciably higher abnormalities than in on-type plants. However, polyvalent chromosomes were observed frequently in both on- and off-type plants in diplotene stage of meiosis. We assume that the unstable morphological characters in resynthesized progeny were the result of abnormal division in meiosis. It results as important that the plants of normal phenotype, chromosome structure and minimized abnormal meiosis are selected to stabilize progeny.

Ascomycetes with cellulolytic, amylolytic, pectinolytic, and mannanolytic activities inhabiting dead beech (Fagus crenata) trees.

It is generally accepted that dead tree decomposition is performed mainly by delignifying basidiomycetes. While ascomycetes have been reported to inhabit dead tree bark, their contribution to dead tree decomposition is still unclear. Here, we isolated five bark-inhabiting ascomycetes possessing cellulolytic activity from dead beech tree and assessed their polysaccharolytic activities. When cultivated in a medium containing filter paper as a sole carbon source, three strains degraded >40 % of the filter paper in a 4-week cultivation and the others degraded 15-30 % of the paper. The degraders possessed amylolytic, pectinolytic, and mannanolytic activities as well as cellulolytic activity, implying that they play an important role in dead tree decomposition after delignification by basidiomycetes. Phylogenetic analysis based on large subunit ribosomal DNA (lsu-DNA) sequences implied that the isolates belonged to Penicillium or Amorphotheca.

Evaluation of patient dose for barium enemas and CT colonography in Japan.

The objective of this study was to assess patient doses for examinations of the lower digestive tract (barium enemas and CT colonography) in Japan. These doses were evaluated from in-phantom dose measurements using a dosemeter-implanted anthropomorphic phantom and from the knowledge of procedures of these examinations. For barium enemas, the doses, which were the sums of doses for various projections in the procedure, were separately derived for fluoroscopy and for analogue and digital radiography. For CT colonography, the doses were evaluated for the prone and the supine positions, each including the doses by scout imaging, and a single abdominal scan for routine and low-dose set-ups. For barium enemas, maximum local skin doses were less than 100 mGy despite relatively long average fluoroscopy times of 8 min; organ doses ranged from 9-26 mGy in the abdomen. The effective dose of 10.7 mSv for analogue radiography decreased by 12% when digital radiography was used, although more than 80% of the dose was due to fluoroscopy. In routine CT colonography performed using a relatively high mean effective mAs of 119 for the accurate detection of colorectal cancer and extra colonic lesions, organ doses within the primary X-ray beam were between 30 mGy and 44 mGy for paired scans whereas, in a low-dose set-up with an effective mAs of 27, they were approximately 10 mGy. Effective doses for routine and low-dose CT colonography of 23.4 mSv and 5.7 mSv were about double and half of the doses for barium enemas, respectively.

Degradation of mikan (Japanese mandarin orange) peel by a novel Penicillium species with cellulolytic and pectinolytic activity.

AIMS: The mikan, or Japanese mandarin orange, is a popular fruit in Japan, but its peel is one of the major agricultural wastes. The aims of this study were to screen, isolate, and characterize a mikan peel-degrading microbe. METHODS AND RESULTS: Several samples including activated sludge, sediment, compost and spoiled mikan peel were collected and cultured in a minimal salt medium containing mikan peel as the sole carbon source. Degradation activity was found in a culture of the spoiled mikan peel, and a fungal strain, designated OP1, with both cellulolytic and pectinolytic activity was isolated. No toxic metabolites, such as mycotoxins, were found in OP1 cultures, as evaluated by gas chromatography/mass spectrometry. A phylogenetic analysis strongly suggested that OP1 is a novel species of the genus Penicillium. CONCLUSIONS: Results suggest that Penicillium sp. OP1 plays an important role in aerobic microbial degradation of cellulose/pectin-rich biomasses in soil ecology, and further imply that this strain may be useful for both simultaneous cellulase/pectinase production and reduction of agricultural waste. SIGNIFICANCE AND IMPACT OF THE STUDY: The present results advance our understanding of microbial degradation of cellulose/pectin-rich biomasses in the natural environment, and offer a new tool for reduction of agricultural waste, which is important for sustaining circulatory societies.

High concentration of deoxycholic acid abrogates in vitro transformation of IEC6 intestinal cells by azoxymethane.

In this study, we designed an in vitro azoxymethane (AOM)-induced carcinogenesis model and analyzed the effect of deoxycholic acid (DCA) on growth, apoptosis, genotoxicity, and transformation of IEC6 intestinal cells. CYP2E1 production was confirmed in IEC6 cells. The growth of IEC6 cells was enhanced by DCA (100 microg/ml). However, IEC6 cells treated with DCA (200 microg/ml) were inhibited and disappeared at 48 hrs after treatment. Apoptotic cells increased 11.2 times by treatment with DCA (200 microg/ml) as compared to cells with no treatment. DNA injury detected by comet assay was found in cells treated with AOM, but not in cells treated with DCA (100 microg/ml) and AOM. The number of colony formation in soft agar increased by AOM treatment. However, the number of foci treated with DCA (100 microg/ml) plus AOM was 69% that of cells treated with AOM alone. Two out of the 6 mice subcutaneously injected with AOM-treated IEC6 cells showed tumorigenesis, whereas IEC6 cells treated with DCA (100 microg/ml) plus AOM or DCA (100 microg/ml) alone did not form any tumor. Reduced protein expression of MLH1, Bcl-2 was detected in IEC6 cells treated with DCA (100 microg/ml). Production of Bax, pJNK, TGF-beta, TGFBRI, TGFBRII, and beta-catenin were higher in IEC6 cells treated with DCA (100 microg/ml) than that in cells with no treatment. These results suggest that high-dose DCA induced apoptosis and inhibited AOM-induced in vitro transformation of IEC6 cells.

Effect of low- and high-intensity pulsed ultrasound on collagen post-translational modifications in MC3T3-E1 osteoblasts.

Different intensities of pulsed ultrasound have distinct biological effects on bone mineralization in the process of bone fracture repair, even across a narrow range (e.g., 30-120 mW/cm(2)). The aim of our study was to elucidate the effect of low-intensity (30 mW/cm(2)) and high-intensity (120 mW/cm(2)) pulsed ultrasound on collagen metabolism by using MC3T3-E1 osteoblasts. Of special interest was the relationship between posttranslational collagen quality and prostaglandin E(2) activity. Cells with or without a cyclooxygenase-2 inhibitor, NS398, were exposed every day for four consecutive days to high-level or low-level intensities of pulsed ultrasound. We examined the, expression patterns of cyclooxygenase-2, lysyl oxidase, telopeptidyl lysyl hydroxylase (TLH), and helical lysyl hydroxylase by real-time polymerase chain reaction analysis. Quantitative analyses of reducible immature and nonreducible mature cross-links were also performed. Ultrasound at 30 mW/cm(2) upregulated TLH messenger RNA (mRNA) expression and enzyme activity compared to the control and resulted in increased relative concentrations of telopeptidyl hydroxylysine-derived cross-links. In addition to upregulated lysyl oxidase mRNA expression, increased total reducible and nonreducible cross-links were observed by 30 mW/cm(2) exposure compared to the control. In contrast, ultrasound at 120 mW/cm(2) had no obvious effect on collagen metabolism owing to high levels of endogenous prostaglandin E(2) induced by ultrasound. Our results showed that (1) low-intensity, but not high-intensity, ultrasound may accelerate the formation of the unique molecular packing of collagen fibers conducive to bone mineralization and that (2) the high dose of endogenous prostaglandin E(2) induced by pulsed ultrasound may be detrimental to calcifiable cross-link formation.

Antibacterial action of several tannins against Staphylococcus aureus.

We examined the antibacterial action of several tannins on plasma coagulation by Staphylococcus aureus and the effect of conventional chemotherapy combined with tannic acid below the MIC. Coagulation was inhibited in plasma containing tannic acid (100 mg/L), gallic acid (5000 mg/L), ellagic acid (5000 mg/L), (-)-epicatechin (1500 mg/L), (-)-epicatechin gallate (500 mg/L) or (-)-epigallocatechin gallate (200 mg/L) after incubation for 24 h. All tannins inhibited coagulation at a concentration below the MIC. The MICs of oxacillin and cefdinir for S. aureus were reduced to < or = 0.06 mg/L in Mueller-Hinton agar plates with tannic acid (100 mg/L) at a concentration below the MIC. The antistaphylococcal activity of tannic acid was reduced in plates with 10% rabbit blood, but not in those with 10% rabbit plasma. Membranous structures formed in a culture medium containing equal proportions of plasma and tryptic soy broth after incubation for 24 h. The colony counts of S. aureus in membranous structures in the medium containing oxacillin (40 mg/L) and tannic acid (100 mg/L) were c. 10-fold lower than those in medium containing oxacillin (40 mg/L) alone (P < 0.01). Tannic acid merits further investigation as a possible adjuvant agent against S. aureus skin infections treated with beta-lactam antibiotics.

[A case of central retinal artery occlusion after anterior posterior fusion of the lumbar spine].

We report a case of central retinal artery occlusion after anterior-posterior fusion of the lumbar spine. The patient suddenly lost his vision of the right eye in the ICU just after the end of long procedure for anterior-posterior fusion of the lumbar spine. The patient was diagnosed as having central retinal artery occlusion, and treated successfully with treatments including immediate administration of urokinase and PGE1, stellate ganglion block, and hyperbaric oxygen therapy. The patient was discharged from the hospital on the 54th postoperative day with adequate vision to drive a car. Central retinal artery occlusion is a rare but very serious complication during and after supine surgery with prone position. It is very important for us to be aware of its possible occurrence. We have to diagnose and treat, as soon as possible, the vision loss after the spine surgery.

Bilateral crossed cerebello-cerebral diaschisis and mutism after surgery for cerebellar medulloblastoma.

A 7-year-old boy developed mutism after surgery for cerebellar medulloblastoma. Postoperative magnetic resonance imaging (MRI) showed atrophy of the cerebellar vermis and both cerebellar hemispheres, predominantly on the right side. Single photon emission computed tomography (SPECT) with technetium-99m-ethyl cysteinate dimer (Tc-99m ECD) revealed decreased cerebral blood flow (CBF) in the bilateral thalami, bilateral medial frontal lobes, and left temporal lobe in addition to the cerebellar vermis and both cerebellar hemispheres when mutism was manifest, indicating the existence of bilateral crossed cerebello-cerebral diaschisis (BCCCD). Circulatory disturbance in both cerebellar hemispheres secondary to tumor resection probably caused BCCCD in both cerebral hemispheres, predominantly in the left, via the dentatothalamocortical pathway (DTCP). With recovery of his mutism, CBF increased in the right thalamus, bilateral medial frontal lobes and left temporal lobe. Thus BCCCD was improved, with only a slight decrease in CBF still persisting in the left thalamus. The mechanism of mutism may have involved damage to the cerebellar vermis (the site of incision at operation), the left dentate nucleus (heavily infiltrated by the tumor) and the right dentate nucleus of the cerebellum (affected by circulatory disturbance secondary to acute postoperative edema). The SPECT findings suggested that mutism was associated with BCCCD-induced cerebral circulatory and metabolic hypofunction in the supplementary motor area mediated via the DTCP.

Chemopreventive effect of curcumin on N-nitrosomethylbenzylamine-induced esophageal carcinogenesis in rats.

Modifying effects of curcumin (derived from the rhizome of Curcuma longa L.) during the initiation or post-initiation phase of N-nitrosomethylbenzylamine (NMBA)-induced esophageal carcinogenesis were investigated in male F344 rats. Five-week-old rats were divided into 5 groups, and groups 1, 2 and 3 were given intraperitoneal injections of NMBA (0.5 mg / kg body weight / injection 15 times) for 5 weeks from 7 weeks old to induce esophageal neoplasms. Groups 2 and 3 were fed the diet containing 500 ppm curcumin during the initiation and post-initiation phases, respectively. Group 4 was given the diet containing curcumin throughout the experiment, and group 5 was kept on the basal diet alone and served as an untreated control. Incidence and multiplicity of esophageal neoplasms of group 1 (NMBA alone) were 66.7% and 0.83 +/- 0.70, respectively. Those of groups 2 and 3 were significantly less than those of group 1 (39.3%, 0.46 +/- 0.64, P < 0.05; 33.3%, 0.36 +/- 0.56, P < 0.05, respectively). Furthermore, the incidence and multiplicity of esophageal preneoplastic lesions (moderate or severe epithelial dysplasia) of group 2 (57.1%, 0.61 +/- 0.57; 40%, 0.29 +/- 0.46) or 3 (56.7%, 0.67 +/- 0.66; 23.3%, 0.23 +/- 0.43) were less than those of group 1 (100%, 1.67 +/- 0.70; 70.8%, 0.92 +/- 0.72) (P < 0.05). In this experiment, feeding of curcumin significantly decreased the expression of cell proliferation biomarkers (5-bromo-2'-deoxyuridine labeling index) in the non-lesional esophageal epithelium (P < 0.01). These findings indicate that curcumin inhibits NMBA-induced esophageal carcinogenesis when given during the post initiation as well as initiation phase. This inhibition may be related to suppression of the increased cell proliferation induced by NMBA in the esophageal epithelium.

Chemopreventive effects of coffee bean and rice constituents on colorectal carcinogenesis.

Polyphenolic compound chlorogenic acid (CGA) known to be much contained in coffee beans was found to have a regressive effect on induced aberrant crypt foci (ACF) as well as on development of ACF in azoxymethane (AOM)-induced colorectal carcinogenesis in rats. Rice germ and gamma-aminobutyric acid-enriched defatted rice germ inhibited AOM-induced ACF formation and colorectal carcinogenesis in rats. Ferulic acid (FA) also known to be contained in coffee beans and rice prevented AOM-induced ACF formation and intestinal carcinogenesis in rats. Both of food factors, coffee and rice may be of benefit to prevention of human colorectal cancers.

Tetrahydrobiopterin-responsive phenylalanine hydroxylase deficiency.

Serum phenylalanine concentrations decreased in 4 patients with hyperphenylalaninemia after loading with tetrahydrobiopterin. There were no abnormalities in urinary pteridine excretion or in dihydropteridine reductase activity. However, mutations were detected in the phenylalanine hydroxylase gene, suggesting a novel subtype of phenylalanine hydroxylase deficiency that may respond to treatment with cofactor supplementation.

Clinical and biochemical aspects of thiamine treatment for metabolic acidosis during total parenteral nutrition.

We encountered six cases of total parenteral nutrition (TPN)-associated lactic acidosis during the 6-y period of 1988-1993. The patients were characterized by severe disease of the digestive organs, minimal food intake before surgery, and postoperative TPN with no food intake and with no vitamin supplements. Within 4 wk of TPN, they developed hypotension (< or = 80/60 mmHg), Kussmaul's respiration, and clouding of consciousness, as well as abdominal pain not directly related to the underlying disease. Routine laboratory examinations revealed no acute aggravation in hepatic, renal, or pancreatic functions. Arterial blood gas analysis showed pH < or = 7.134 and base excess < or = -17.5 mmol/L. Additional laboratory examinations revealed serum lactate > or = 10.9 mmol/L, serum pyruvate > or = 159 mumol/L, and lactate/pyruvate ratio > or = 0.029. None of the patients responded to sodium bicarbonate or other conventional emergency treatments for shock and lactic acidosis. After the first case, we suspected that thiamine deficiency might be responsible for this pathologic condition, Serum thiamine was proved to be < or = 196 nmol/L in 5 patients. Thiamine replenishment at intravenous doses of 100 mg every 12 h resolved lactic acidosis and improved the clinical condition in 3 patients. This article includes a review of 11 relevant reports published from 1982-1992 and a discussion of the biochemical mechanism of onset of thiamine deficiency-associated lactic acidosis. We emphasize the needs (1) to supplement TPN with thiamine-containing vitamins for the patients whose food intake does not meet nutritional requirements; (2) to monitor the patients routinely measuring serum thiamine concentration and erythrocyte transketolase activity during TPN; and (3) to intravenously replenish using high-dose thiamine simultaneously with the manifestation of signs and symptoms of lactic acidosis.

Multiple regression analysis of relationship between frontal lobe phosphorus metabolism and clinical symptoms in patients with schizophrenia.

We investigated the differences among diagnostic types of 36 schizophrenic patients in the brain phosphorus metabolism in the frontal lobe. We performed phosphorus-31 magnetic resonance spectroscopy (31P-MRS) in the frontal region in patients with schizophrenia of the catatonic (n = 4), disorganized (n = 8), paranoid (n = 10) and undifferentiated (n = 14) types. In the disorganized type, the PME level was significantly decreased compared to those in the other three types, while the phosphodiester (PDE) level tended to be higher, although not significantly, than those in the other types. Using multiple regression analysis, we investigated whether or not the clinical symptoms were correlated with the brain phosphorus metabolism. An increased motor retardation factor score was significantly correlated with decreased PME level, whereas more severe emotional withdrawal and blunted affect were associated with increased PDE level. These results suggest that altered membrane phospholipid metabolism in the frontal region may be associated with negative symptoms and that schizophrenia of the disorganized type is associated with more severe negative symptoms and may present more severe brain abnormalities compared to the other types.

Effects of nilvadipine (a dihydropyridine-type calcium entry blocker) on cerebral blood flow in acute experimental brain ischemia in rats.

The purpose of the present study was to examine the effect of nilvadipine (a dihydropyridine-type calcium entry blocker) on the cerebral circulation, experimental models of cerebral ischemia were used to measure abrupt changes in the cortical cerebral blood flow (CBF) at the 'penumbra' (the boundary region between the anterior cerebral artery (ACA) and the middle cerebral artery (MCA)). Left middle cerebral artery occlusion (MCAO) was performed in Sprague--Dawley rats, and then 8 or 16 micrograms kg-1 of nilvadipine was administered to two groups of the rats. The former group was referred to as the 8-MCAO(+) group (n = 8), and the latter group, the 16-MCAO(+) group (n = 8). To a control group (n = 8), only the solvent of nilvadipine was administered. After the administration in each group, CBF and the mean arterial blood pressure (MAP) were continuously measured for 60 min. The MAP in both the 8-MCAO(+) and 16-MCAO(+) groups was significantly lower than in the control group for 25 min after nilvadipine administration (Wilcoxon 2 sample test, p < 0.05). Also, the CBF in the 16-MCAO(+) group was significantly lower (about 20%) for 55 minutes than that in the control group. On the other hand, the CBF in the 8-MCAO(+) group three minutes after nilvadipine administration was not significantly lower than in the control group (Wilcoxon 2 sample test). Our findings confirmed that nilvadipine (8 micrograms kg-1) maintained CBF while lowering the blood pressure in the experimental model of acute cerebral ischemia.

Reactivation of phototoxicity test for psoralens plus ultraviolet A by low-dose methotrexate.

Reactivation of a solar burn has been reported after administration of methotrexate for cancer and psoriasis patients. We report a case of psoriasis with a similar but delayed elicitation of phototoxic reaction by administration of low-dose methotrexate one month after psoralen and long-wave ultraviolet light (PUVA) treatment. The reactivation was seen in an already receding phototoxicity test area but not in a less phototoxic, noninterval PUVA treated area, suggesting that the reactivation by methotrexate is related with the severity of light-induced skin inflammation.

Micro-PIXE (particle induced X-ray emission) analysis of aluminum in rat-liver using MeV heavy ion microprobes.

Heavy ion microprobes (HIM) such as 3 MeV Si2+ and 3 MeV p2+ have been applied to the elemental analysis by PIXE (proton-induced X-ray emission). It was found that silicon and phosphorus microprobes have several times higher sensitivity for aluminum K alpha X-rays than 2 MeV proton microprobes, and detection limits were more favorable in a phosphorus microprobe. Using a 3 MeV P2+ microprobe, the liver of a rat, which had been injected with aluminum-lactate, was investigated and it was found that aluminum segregates in areas with a dimension of about 10 microns. These areas could hardly be observed with 2 MeV proton microprobes.

Inhibitory effect of gomisi on reductive metabolism of halothane.

The effect of Gomisi (dried ripe fruit of schizandra chinensis) on chlorodifluoroethylene (CDE) and chlorotrifluoroethane (CTE) formation was investigated. The incubation mixtures for the measurement of reductive metabolites of halothane consisted of liver microsomal suspensions, 3 mM NADPH, extract solution of Gomisi and halothane in 0.1 M potassium phosphate buffer (pH 7.4). The production of CDE and CTE was inhibited by Gomisi in a dose-dependent way. The production were reduced to half in the presence of 0.5% Gomisi extract in the reaction mixture. The results suggest that Gomisi can inhibit the reductive metabolism of halothane in vitro; thus it may protect against halothane-induced hepatitis.

Chemotherapeutic efficacy of a newly synthesized benzoxazinorifamycin, KRM-1648, against Mycobacterium avium complex infection induced in mice.

Newly synthesized benzoxazinorifamycin, KRM-1648, was studied for its in vivo anti-Mycobacterium avium complex (MAC) activities. When the MICs were determined by the agar dilution method with Middlebrook 7H11 agar medium, KRM-1648 exhibited similarly potent in vitro antimicrobial activities against the MAC isolated from AIDS and non-AIDS patients, indicating possible usefulness of KRM-1648 against AIDS-associated MAC infections. KRM-1648 exhibited potent therapeutic activity against experimental murine infections induced by M. intracellulare N-260 (virulent strain) and N-478, which has much weaker virulence. Similarly, KRM-1648 exhibited an excellent therapeutic efficacy against M. intracellulare infection induced in NK-cell-deficient beige mice (as a plausible model for AIDS-associated MAC infection), in which a much more progressed state of gross lesions and bacterial loads at the sites of infection were observed. When the infected beige mice were killed at weeks 4 and 8, obvious therapeutic efficacy was seen on the basis of reduction in the incidence and degree of lung lesions and bacterial loads in the lungs and spleen with infections due to M. intracellulare N-241, N-256, and N-260. In this case, the efficacy was the highest in N-260 infection, followed by strain N-241. When mice were observed until infection-induced death, survival time of the infected beige mice was found to be prolonged by KRM treatment. However, KRM-1648 was not efficacious in suppressing the progression of pulmonary lesions and the increase in bacterial loads at the sites of infection, including lungs and spleen, at the late phase of infection. This may imply some difficulty with chemotherapy for AIDS-associated MAC infection, even with KRM-1648 treatment, which has excellent in vitro and in vivo anti-MAC activities, as shown in present study.