Diversity and Distribution of Rice Blast (Pyricularia oryzae Cavara) Races in Japan.

In total, 310 rice blast (Pyricularia oryzae Cavara) isolates from Japan showed wide variation in virulence. Virulence on rice (Oryza sativa L.) differential varieties (DV) harboring resistance genes Pish, Pia, Pii, Pi3, Pi5(t), Pik-s, and Pi19(t) ranged from 82.9 to 100.0%. In contrast, virulence on DV possessing Pib, Pit, Pik-m, Pi1, Pik-h, Pik, Pik-p, Pi7(t), Pi9(t), Piz, Piz-5, Piz-t, Pita-2, Pita, Pi12(t), and Pi20(t) ranged from 0 to 21.6%. Cluster analysis using the reaction patterns of the DV classified isolates into three groups: I, virulent to Pik, Pik-h, Pik-p, Pik-m, Pi1, and Pi7(t); IIa, avirulent to the preceding 6 genes and virulent to Pia, Pii, Pi3, and Pi5(t); and IIb, avirulent to all 10 genes. Group I was limited to northern Japan and group IIb to central Japan, while group IIa was distributed throughout Japan. We estimate that group IIa represents the original population and that groups I and IIb arose from it through minor changes in pathogenicity. We classified these isolates into 123 races by a new designation system and conclude that the rice blast races in Japan are less diverse than previously thought.

Diversity and Distribution of Rice Blast (Pyricularia oryzae Cavara) Races in Bangladesh.

The pathogenicity of 331 blast isolates (Pyricularia oryzae Cavara) collected from different regions and ecosystems for rice cultivation in Bangladesh was evaluated by compatibility on 23 differential varieties (DV), each harboring a single blast resistance gene, and susceptible 'Lijiangxintuanheigu' (LTH). A wide variation in virulence was found among the isolates, and 267 races were classified using a new designation system. Virulence of blast isolates against DV carrying the resistance genes Pia, Pib, Pit, Pik-s, Piz-t, Pi12(t), Pi19(t), and Pi20(t), as well as avirulence against those carrying Pish, Pi9, Pita-2, and Pita, was distributed widely in Bangladesh. Cluster analysis of the compatibility data on the DV initially classified the isolates into groups I and II. The virulence spectra of the two groups differed mainly according to the reactions of the DV to Pii, Pi3, Pi5(t), Pik-m, Pi1, Pik-h, Pik, Pik-p, and Pi7(t). Group I isolates were distributed mainly in rainfed lowlands, whereas group II isolates were found mainly in irrigated lowlands; however, there were no critical differences in geographic distribution of the blast isolates. In total, 26 isolates, which could be used to identify the 23 resistance genes of the DV on the basis of their reaction patterns, were selected as a set of standard differential blast isolates. To our knowledge, this is the first clear demonstration of the diversity and differentiation of blast races in Bangladesh. This information will be used to develop a durable blast protection system in that country.

Evaluation by dental cone-beam computed tomography of the incidence and sites of branches of the inferior dental canal that supply mandibular third molars.

Our aim was to assess the incidence and anatomical site of branches of the inferior dental canal that supply mandibular third molars using dental cone-beam computed tomography (CT). We evaluated the incidence and diameter of branches of the inferior dental canal using 272 cone-beam CT mandibular scans from 172 patients referred for imaging before the extraction of impacted mandibular third molars. We found three typical branching patterns from the inferior dental canal in the third molar region: the retromolar canal (in the retromolar triangle), the dental canal (that courses directly beneath the socket of the third molar), and the accessory canal (that courses through the socket and leads from the inferior dental canal to a bony ridge). The incidences of retromolar, dental, and accessory canals were 75 (28%), 223 (82%), and 21 (8%), respectively, with mean diameters of 0.9 (0.4), 0.7 (0.5), and 1.1 (0.4) mm, respectively. Operative injury to the neurovascular contents within the branches of the inferior dental canal can lead to excessive bleeding and postoperative paraesthesia, so identification of its branches on preoperative cone-beam CT images may prove useful during extraction of impacted mandibular third molars or when harvesting bone blocks from the region of mandibular third molars. We also describe two cases of branches detected on panoramic and cone-beam CT images that prompted this research.

HeLa ZIP kinase induces diphosphorylation of myosin II regulatory light chain and reorganization of actin filaments in nonmuscle cells.

Dlk/ZIP kinase is a serine/threonine kinase highly homologous to DAP kinase. We have reported that HeLa ZIP kinase (hZIPK) phosphorylated the regulatory light chain of myosin II (MRLC) at both Ser19 and Thr18 in vitro. In this study, we demonstrate that hZIPK also induces the diphosphorylation of MRLC in nonmuscle cells. Peptide mapping revealed that transient transfection of hZIPK into HeLa cells caused diphosphorylation of MRLC. In contrast, transfection of the kinase inactive mutant of hZIPK did not induce any phosphorylation of MRLC. Using antibodies specific for mono- or diphosphorylated MRLC, we showed that diphosphorylated MRLC induced by the overexpression of hZIPK was concentrated in striking aggregates or bundles of actin filaments in HeLa cells, while monophosphorylated MRLC showed no prominent localization to these aggregates. Overexpression of hZIPK also induced dramatic changes in cell shape and disruption of nuclear morphology reminiscent of changes during apoptosis. These effects of hZIPK were suppressed by the coexpression of a mutant MRLC where both phosphorylation sites were replaced with alanine, indicating that the changes in actin organization were a consequence of MRLC diphosphorylation. These results suggested that hZIPK plays a role in regulating actin organization and cell morphology in non-muscles and at least part of its effects are mediated through the diphosphorylation of MRLC.

Effect of angiotensin II on Ca2+ efflux from freshly isolated adult rat cardiomyocytes: possible involvement of Na+/Ca2+ exchanger.

In the present study, we examined the effect of angiotensin II on Ca2+ efflux from freshly isolated adult rat cardiomyocytes. Angiotensin II stimulated the efflux of 45Ca2+ from the cells in a concentration-dependent manner, at least in pharmacological doses of 10(-8) M to 10(-5) M. The 45Ca2+ efflux was inhibited by the type 1 angiotensin II receptor antagonist losartan, but not by the type 2 antagonist PD 123319. Angiotensin II also induced an increase in cytosolic free calcium ([Ca2+]i) and inositol trisphosphate formation within the cardiomyocytes. Angiotensin II-induced 45Ca2+ efflux and the increase in [Ca2+]i were both inhibited by thapsigargin, a specific inhibitor of the sarcoplasmic reticulum Ca2+ pump. The 45Ca2+ efflux was not affected by removal of the extracellular Ca2+ but was dependent on the presence of extracellular Na+. In addition, angiotensin II caused 22Na+ influx into the cells. These results indicate that angiotensin II stimulates Na+-dependent 45Ca2+ efflux from freshly isolated adult rat cardiomyocytes, probably through its stimulatory effect on the plasma membrane type 1 angiotensin II receptors. Angiotensin II-induced increase in [Ca2+]i may cause an activation of Na+/Ca2+ exchange which finally results in the stimulation of 45Ca2+ efflux from the cells. Since it is reported that Na+/Ca2+ exchange is important in calcium homeostasis within the cells, angiotensin II may play some role in the reduction of intracellular Ca2+ from isolated adult rat cardiomyocytes.

Myosin II regulatory light chain as a novel substrate for AIM-1, an aurora/Ipl1p-related kinase from rat.

Previous studies demonstrated that the phosphorylated myosin II regulatory light chain (MRLC) is localized at the cleavage furrow of dividing cells, suggesting that phosphorylation of MRLC plays an important role in cytokinesis. However, it remains unclear which kinase(s) phosphorylate MRLC during cytokinesis. AIM-1, an Aurora/Ipl1p-related kinase from rat, is known as a serine/threonine kinase that is required for cytokinesis. Here we examined the possibility that AIM-1 is a candidate for a kinase that phosphorylates MRLC during cytokinesis. As a result, we showed that AIM-1 monophosphorylated MRLC at Ser19 using two-dimensional phosphopeptide mapping analysis and several MRLC mutants. Furthermore, AIM-1 was colocalized with monophosphorylated MRLC at the cleavage furrow of dividing cells. We propose here that AIM-1 may participate in monophosphorylation of MRLC during cytokinesis.

Antiplatelet effect of Z-335, a new orally active and long-lasting thromboxane receptor antagonist.

We investigated the pharmacological characteristics of Z-335 ((+/-)-sodium[2-[4-(chlorophenylsulfonylaminomethyl)indan-5-yl]ace tate monohydrate), a new indan derivative. Z-335 inhibited the specific binding of [3H]SQ-29548 to human platelets and guinea pig platelet membranes. The IC50 values of Z-335 for human platelets and guinea pig platelet membranes were 29.9 +/- 3.1 nM with a slope of 1.09 +/- 0.05 and 32.5 +/- 1.7 nM with a slope of 1.07 +/- 0.02, respectively. Z-335 inhibited thromboxane A2 receptor-mediated human and guinea pig platelet aggregation in vitro and oral administration of this drug to guinea pigs inhibited U-46619- and collagen-induced platelet aggregation for 24 h. Z-335 dose-dependently prevented the occurrence of U-46619-induced pulmonary thromboembolism in mice and the protective effect of this drug (0.3 and 3 mg/kg, p.o.) lasted for 24 h. These results strongly suggest that Z-335 is a potent, orally active and long-lasting thromboxane A2 receptor antagonist, which may be useful as an antiplatelet drug.

Pharmacodynamic analysis of steroid 5alpha-reductase inhibitory actions of Z-350 in rat prostate.

The pharmacodynamics of (S)-4-[3-[4-[1-(4-methylphenyl)-3-[4-(2-methoxyphenyl)piperazine-1-yl]propoxy]benzoyl]indole-1-yl] butyric acid hydrochloride (Z-350), which has alpha(1)-adrenoceptor antagonistic and steroid 5alpha-reductase inhibitory effects, were investigated in rats. The disposition of Z-350 was a function of linear kinetics at doses from 1 to 30 mg/kg; the bioavailability was calculated to be 65.2%. The inhibition of 5alpha-reductase was dependent on the concentration of Z-350 in plasma and in the prostate. Analysis of the relationship between the concentration in the prostate and the inhibition seen after a single oral administration showed that the Hill constant was almost 1.0 and EC(50)(n(H)) was 47.4 ng/g of tissue; these parameters did not change after multiple administration. Z-350 inhibited 5alpha-reductase 1 h after oral administration at a dose of 3 mg/kg; maximum inhibition was observed after 2-4 h, and the inhibition (%) was maintained for 24 h after oral administration.

Angiotensin II as a stimulator of Na(+)-dependent Ca2+ efflux from freshly isolated adult rat cardiomyocytes.

In cardiac tissues, angiotensin II causes inotropic and chronotropic effects on the heart. It is indicated that the mechanism of the inotropic effect of angiotensin II is attributed to an increase in cytosolic free calcium ([Ca2+]i) in cardiomyocytes. However, increased [Ca2+]i should be restored to a physiological level because cumulative elevation in [Ca2+]i leads to irreversible injury in cardiomyocytes. Whereas it is known that angiotensin II causes the increase in [Ca2+]i in cardiac cells, little is known about the mechanisms of decrease in [Ca2+]i in cardiomyocytes upon angiotensin II stimulation. In the present study, we examined the effect of angiotensin II on Ca2+ efflux from freshly isolated adult rat cardiomyocytes. Angiotensin II stimulated the efflux of 45Ca2+ from the cells in a concentration-dependent manner (10(-7)-10(-5) M). The 45Ca2+ efflux from the cells was inhibited by type 1 angiotensin II receptor inhibitor. The angiotensin II-stimulated 45Ca2+ efflux was not affected by deprivation of the extracellular Ca2+, but was dependent on the presence of extracellular Na+. These results indicate that angiotensin II stimulates extracellular Na(+)-dependent 45Ca2+ efflux from freshly isolated adult rat cardiomyocytes, probably through its stimulatory effect on the plasma membrane type 1 angiotensin II receptors which may couple to Na+/Ca2+ exchange.

A new thromboxane receptor antagonist, Z-335, ameliorates experimental thrombosis without prolonging the rat tail bleeding time.

We investigated the antithrombotic activity of Z-335, a new thromboxane A2 receptor antagonist, using experimental thrombosis models, and also tested its effect on the rat tail bleeding time. Z-335 (0.1, 0.3, and 1 mg/kg, p.o.) dose-dependently prevented the occurrence of arachidonic acid-induced pulmonary thromboembolism in mice. During photochemically induced thrombosis in the femoral artery of guinea pigs, Z-335 (0.3, 1, and 3 mg/kg, i.v.) dose-dependently prolonged the time required to form thrombi. Moreover, Z-335 (10 mg/kg/day, p.o.) strongly suppressed lauric acid-induced hind limb injury in rats. Z-335 (0.3, 3, 30, and 300 mg/kg, p.o.) did not prolong the tail bleeding time in rats. These results strongly suggest that Z-335 ameliorates experimental thrombosis without prolonging the rat tail bleeding time, and may therefore be a useful antithrombotic drug.

Intraosseous neurilemmoma of the mandible.

We report a rare case of intraosseous neurilemmoma of the mandible, with an emphasis on radiographic findings. The tumor, located mainly in the premolar region, presented as an expansive, unilocular, well-defined, radiolucent lesion on plain radiography. No dilatation of the mandibular canal was identified. MR imaging helped to identify the solid nature of the tumor. A biopsy was necessary to make the final diagnosis because of the relatively nonspecific nature of the lesion.

Z-350, a new chimera compound possessing alpha1-adrenoceptor antagonistic and steroid 5alpha-reductase inhibitory actions.

The effects of Z-350, (S)-4-[3-(4-{1-(4-methyl-phenyl)-3-[4-(2-methoxyphenyl)piperazine-1-yl]propoxy} benzoyl)indole-1-yl]butyric acid hydrochloride, a newly synthesized compound possessing alpha-adrenoceptor antagonistic and steroid 5alpha-reductase inhibitory actions, were studied in vitro. In functional experiments, Z-350 shifted the concentration/response curve for the phenylephrine-induced contraction of rabbit prostate, urethra and aorta to the right with pA2 values of 8.04, 7.57 and 7.13, respectively. The binding affinity of Z-350 for alpha1-adrenoceptors in rabbit prostate, urethra and aorta were estimated by the displacement of [3H]prazosin. The pKi values for this action of Z-350 were 7.53, 7.95 and 7.62 for the prostate, urethra and aorta, respectively. alpha1-Adrenoceptor subtype selectivities were studied in the submaxillary gland (r(1A) and liver (alpha1B) of rat. Z-350 inhibited the specific binding of [3H]prazosin to alpha1A and (alpha1B-adrenoceptors with pKi values of 7.82 and 7.29, respectively. Z-350 inhibited rat prostatic steroid 5alpha-reductase non-competitively with a pIC50 of 8.42. These results indicate that Z-350 is a alpha1-adrenoceptor antagonist and is a steroid 5alpha-reductase inhibitor. It is expected that Z-350 will be a candidate drug for the treatment of benign prostatic hyperplasia.

Treatment results of oral verrucous carcinoma and its biological behavior.

The biologic behavior of and optimal treatment for oral verrucous carcinoma (VC) remain controversial. We analyzed the clinicopathological characteristics of 12 patients with oral VC. Immunohistochemical techniques were used to evaluate p53 protein, CD44 variant 9, and proliferating cell nucleus antigen. The TNM classification (UICC, 1997) was T1 in 1 patient, T2 in 3, T3 in 4, and T4 in 4. All patients were classified as N0M0. Four patients were treated by surgery alone and 8 by surgery after chemotherapy, radiotherapy, or both. After surgery, two patients had primary recurrence of disease. Immunohistochemically, the proliferative activity of tumor cells as evaluated by proliferating cell nuclear antigen labeling index and p53 protein expression was similar in VC and well-differentiated squamous cell carcinoma. However, CD44 varient 9 expression was positive in 8 of 10 VC, suggesting that oral VC is associated with a low risk of lymph node metastasis. Positive CD44 variant 9 expression by most oral VCs, indicating a low risk of cervical lymph node metastasis, suggests that most cases can be controlled by surgical intervention.

Utilization of cell response under heat, chemical, and combined stresses for selective recovery of cytoplasmic beta-galactosidase from Escherichia coli cells.

The effective bioseparation process was developed by exploiting the cell response under heat, chemical, and combined stresses for selective recovery of cytoplasmic beta-galactosidase from Escherichia coli cells. At the observed optimal condition for heat stress (45 degrees C, 30 min) and that for chemical stress (20 mM Triton X-100, 5 mM EDTA, 30 min), E. coli cells were exposed to these stresses either simultaneously or sequentially. The operational sequence highly affected the yield and selectivity of the target recovery. Exposing the cells to the sequential stress (heating at 45 degrees C for more than 30 min after Triton X-100/EDTA treatment for 30 min) was found to be most effective for selective recovery of target beta-galactosidase among possible sequences.

Surgical management of infants with mitral valve stenosis or atresia without diminutive ascending aorta.

The surgical strategy in infants with mitral valve stenosis or atresia without diminutive ascending aorta remains to be established, including the potential for biventricular repair as a definitive operation. Our surgical experience of six infants with mitral valve stenosis (4 patients) or atresia (2 patients) without diminutive ascending aorta was evaluated based on three important factors: left ventricular volume; the nature of the systemic outflow obstruction; and the type of mitral valve anomaly. Two patients with systemic outflow tract diameter less than 65% of normal underwent systemic outflow tract reconstruction, and the other patients with outflow tract diameter more than 68% of normal were able to maintain systemic circulation without repair. Only one patient with mitral valve stenosis without left ventricular outflow tract obstruction underwent a successful open mitral valvotomy as a biventricular repair after first-stage palliation. The left ventricle of the other patients did not grow after first-stage palliation. Due to progressive subaortic narrowing, pulmonary artery banding should be avoided in patients with mitral atresia due to absent atrioventricular connection who are future Fontan candidates. Most patients with this lesion can be expected to become candidates for safe Fontan-type repair.

Effects of intravenous perilla oil emulsion on nutritional status, polyunsaturated fatty acid composition of tissue phospholipids, and thromboxane A2 production in streptozotocin-induced diabetic rats.

The effects of a perilla oil (PO) emulsion rich in alpha-linolenic acid, administered by intravenous infusion, on nutritional status, fatty acid composition, and thromboxane A2 production were compared with those of a soybean oil (SO) emulsion in streptozotocin-induced diabetic rats given a fat-free diet for 7 days. The PO emulsion improved body weight gain and nitrogen balance compared with the SO emulsion and reduced thromboxane A2 production by platelets. The PO emulsion also increased the proportion of eicosapentaenoic acid, but decreased that of arachidonic acid, in liver and serum phospholipids. Plasma insulin concentrations and blood biochemical indices were similar in the two groups. An intravenously infused PO emulsion effectively reduces thromboxane A2 production through changes in the fatty acid composition of liver and serum phospholipids, as with oral administration, and improves the nutritional status of diabetic rats.

Preparation of fine emulsified fat particles without glycerol for intravenous nutrition.

A method of preparing fine emulsified fat particles without glycerol for intravenous nutrition was investigated. The factors assessed were the oil phase ratio, the glucose level of the aqueous phase and the temperature of high-pressure homogenization. The particle size decreased with an increase in the oil phase ratio and it went below 250 nm only in the emulsion with a 50% oil phase ratio. The weight-weighted particle size (dw)/number-weighted particle size (dn) value reflected the particle size distribution. The emulsion with a 50% oil phase ratio had a very narrow distribution of particle sizes and the dw/dn value was below 1.1. With the use of glucose solutions for the aqueous phase, smaller particle sizes and narrower distributions were obtained with increasing glucose concentrations. The controlled temperature of 50 degrees C was appropriate for high-pressure homogenization, producing particles below 160 nm. The rate of the layer separation was a function of particle size. The particle sizes below 180 nm can be expected to suppress the separation of the formulation which consisted of 10.0% soybean oil, 1.2% phospholipids and 5.0% glucose. The stability studies were conducted at 40 degrees C for 3 months and the fat emulsion was stable during storage. These investigations contribute to the preparation of a new caloric source for peripheral parenteral nutrition.

Long-term restoration of masticatory function with fixed mandibular implants in cases of oral cancer.

The results of the present study show that the fixed mandibular implant system enables patients who have operations to treat oral cancer to use a stabilized denture continuously over an extended period of time.

Direct intraoperative measurements of aortic and pulmonary blood flows in patients with severe pulmonary artery hypertension.

BACKGROUND: Evaluate the significance of direct intraoperative measurements of aortic and pulmonary blood flows by electromagnetic flowmeter as an absolute decision basis for operability in patients with ventricular septal defect/complete atrioventricular septal defect and severe pulmonary artery hypertension. EXPERIMENTAL DESIGN: Prospective study. SETTING: Institutional practice. PATIENTS: Eight patients with marginal operability based on preoperative Doppler echocardiography and cardiac catheterization (pulmonary-to-systemic flow ratio=1.1-2.3, pulmonary-to-systemic resistance ratio=0.34-0.91, and pulmonary vascular resistance=4.6-18.2 units x m2) underwent direct intraoperative measurements of aortic and pulmonary blood flows by electromagnetic flowmeter. Operation would be performed according to the results of direct intraoperative measurements in every patient. RESULTS: Aortic flow by direct intraoperative measurements ranged from 0.9 to 3.2 L/min/m2, and pulmonary blood flow from 4.1 to 8.4 L/min/m2. Pulmonary-to-aortic flow ratio was calculated at 2.1-6.6. Pulmonary vascular resistance ranged from 2.6 to 7.7 units x m2. We assessed that all patients still had operability, and performed corrective operations. Postoperative courses corresponded with the data from the direct intraoperative measurements. CONCLUSIONS: When some clinical findings, particularly Doppler echocardiographic findings, of these patients are slightly in favor of reversibility of pulmonary vascular disease despite discrepant data of preoperative cardiac catheterization under a tight control of carbon dioxide tension, we recommend that direct intraoperative measurement of aortic and pulmonary blood flows is especially useful in decision making for the operability of patients with severe pulmonary artery hypertension.

Correlation between intravenously supplied energy level and zinc metabolism in laparotomized rats.

We investigated the relationship between intravenous energy loading and zinc status in laparotomized rats. One of three test solutions consisting of 3% amino acid, the same amount of electrolytes (excluding zinc) and different concentrations of glucose were infused through the jugular vein for 5 d. The total energy was 109, 191 and 273 kcal/kg/d, respectively. Significantly positive correlations were observed between infusion energy and rat body weight changes (% of initial value) and between infusion energy and cumulative nitrogen balance. Regarding the zinc status, a negative correlation was found between infusion energy and plasma zinc concentration, and a positive correlation was observed between infusion energy and urinary zinc excretion. There was no significant relationship between infusion energy and hepatic zinc content. These results indicate that the zinc requirement might be increased when infusion energy is elevated and the nutritional status is improved. Zinc supplementation in the post-operative period should be considered in light of not only catabolism but also anabolism. Anabolism may be more important than catabolism in regard to zinc metabolism under relatively mild stress.