Characterization of microbial consortia that reductively dechlorinate 4-chlorophenol and transform phenol to benzoate enriched from estuarine sediment of Lake Shinji.

Monochlorophenols were degraded to benzoate via phenol by the initial dechlorination and the subsequent conversion of phenol to benzoate in anaerobic sediment samples of estuarine Lake Shinji under methanogenic conditions. To characterize bacteria that dechlorinate 4-chlorophenol and transform phenol to benzoate, we analyzed the microbial community structure of the enrichment culture with each 4-chlorophenol and phenol by the limiting dilution method with polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) of 16S rRNA gene. After serial dilution of the culture, the 4-chlorophenol-dechlorinating culture consisted of two dominant bacteria, one of which was most homologous with Dehalobacter sp. In the enriched culture with phenol, minor band homologous with Cryptanaerobacter phenolicass corresponded to the transformation activity.

Hygiene measures considering actual distributions of microorganisms in Japanese households.

AIMS: Effective household hygiene measures require that sources of bacterial contamination and the places to which contamination spreads be carefully identified. Therefore, a study was performed to examine the distribution of microorganisms throughout ordinary households in Japan, which has its own unique customs of daily life and food preparation. METHODS AND RESULTS: Using the stamping method, samples were taken from 100 different places and items in each of 86 households. This study found kitchens/dining rooms to have the greatest level of microbial contamination and bathrooms, the next highest level. Toilets (water closets) were found to have an unexpectedly low level of bacterial contamination. The largest bacterial counts were found on items such as drain traps, dish-washing sponges, counter towels, sinks, dish-washing tubs, and bathroom sponges. CONCLUSIONS: It is necessary to carefully identify both the items that can become instruments for spreading bacterial contamination and the places that easily become subject to secondary contamination, and then to take timely and effective disinfection/sanitizing measures. SIGNIFICANCE AND IMPACT OF THE STUDY: The data gathered in this study will be very valuable for anticipating the pathways over which bacteria are transported and prioritizing disinfection targets, to make effective disinfection possible.

Bacterial contamination of Japanese households and related concern about sanitation.

The bacterial contamination of Japanese homes and the attitudes of Japanese people toward sanitation were studied. By taking bacterial counts of approximately 90 places each in five homes, this study found kitchens to have the greatest degree of bacterial contamination, followed by bathing rooms. Toilets had less bacterial contamination than was expected. While concern about bacteria on highly contaminated items such as sponges, towels for wiping counters, and other reservoirs/disseminators was common, there was a relative lack of concern regarding contact surfaces such as dining tables. It is believed that an in-depth study of bacterial contamination in the home and concern about it would lead to the promotion of greater public understanding of home sanitation and help to facilitate the provision of useful information and products to the public.

Antianginal effects of hydroxyfasudil, a Rho-kinase inhibitor, in a canine model of effort angina.

1. The effects of Rho-kinase inhibitor, fasudil, and of a more specific Rho-kinase inhibitor, hydroxyfasudil, on pacing-induced myocardial ischaemia were determined in anaesthetized open-chest dogs. 2. The dogs were subjected to left anterior descending coronary artery (LAD) stenosis producing a sufficient ischaemia as measured by ST-segment depression on electrocardiograms only when the hearts were paced 60 beats min(-1) above the baseline. After a recovery (nonpacing) period, drugs or saline were infused intravenously over 30 min. The animals were again subjected to 5 min of pacing 25 min after the initiation of the treatment. 3. Hydroxyfasudil (0.1 and 0.3 mg kg(-1)) and fasudil (0.3 mg kg(-1)) suppressed the ST-segment depression. Hydroxyfasudil and fasudil also increased the regional blood flow of the LAD perfused endomyocardium region in the canine model of effort angina. 4. To determine the flow profile for hydroxyfasudil in dogs, blood flow in three vascular beds was measured. Hydroxyfasudil (0.3 mg kg(-1) for 30 min) significantly increased coronary blood flow and vertebral blood flow, without significantly changing the femoral blood flow. 5. Hydroxyfasudil had no inotropic or chronotropic effect on the isolated hearts of guinea-pigs. Hydroxyfasudil (2 mg kg(-1) for 20 min) did not affect the PR or QTc interval in anaesthetized dogs. 6. Inhibition of Rho-kinase appears to protect myocardium subjected to pacing-induced ischaemia through the increase in the regional myocardial blood flow. Hydroxyfasudil may be categorized as a novel type of anti-anginal drug, without any inotropic or chronotropic effects.

Risk stratification analysis of operative mortality in heart and thoracic aorta surgery: comparison between Parsonnet and EuroSCORE additive model.

OBJECTIVE: Our purpose was to compare the performance of risk stratification model between Parsonnet and European System for Cardiac Operative Risk Evaluation (EuroSCORE) in our patient database. METHODS: From August 1994 to December 2000, 803 consecutive patients have undergone heart and thoracic aorta surgery using cardiopulmonary bypass and scored according to Parsonnet and EuroSCORE algorithm. The population was divided into five clinically relevant risk categories. We compared correlation of predicted mortality and observed mortality between these two models. Score validity was assessed by calculating the area under the receiver operating characteristic (ROC) curve. RESULTS: Overall hospital mortality was 4.5%. In Parsonnet model, predicted mortality was 2.4% for 0-4% risk, 6.7% for 5-9% risk, 12% for 10-14% risk, 17% for 15-19% risk, 25% for 20% plus risk, and 10.4% for overall patients. Observed mortality was 2.4, 0.4, 5.9, 8.7, 11, and 4.5%, respectively. The thoracic aorta and valve cohort indicated poor correlation between predicted and observed mortality compared to coronary cohort. In the EuroSCORE model, predicted mortality was 1.4% for 0-2% risk, 4.0% for 3-5% risk, 6.7% for 6-8% risk, 9.7% for 9-11% risk, 13% for 12% plus risk, and 5.3% for overall patients. Actual mortality was 0, 1.5, 6.8, 11, 21, and 4.5%, respectively. Each of the thoracic aorta, valve, and coronary cohort indicated good correlation between predicted and observed mortality. Areas under the ROC curves were 0.72 in Parsonnet and 0.82 in EuroSCORE. CONCLUSIONS: The EuroSCORE additive model yielded good predictive value for hospital mortality of Japanese patients undergoing not only cardiac but also thoracic aortic surgery.

Risk stratification analysis of operative mortality in coronary artery bypass surgery.

OBJECTIVE: We assessed the operative mortality of coronary artery bypass grafting (CABG) surgery using risk stratification. METHODS: In 294 consecutive patients who underwent CABG with or without concomitant surgery from August 1994 to December 1999, we compared operative mortality calculated conventionally and by risk stratification. Scores for each patient were calculated using the Parsonnet additive model and stratified based on the probability of operative mortality. RESULTS: Overall crude hospital mortality was 4.8%-4.0% among patients younger than 80 years and 14% among those 80 years of age or older (p = 0.0692). Hospital mortality was 12% in urgent/emergency surgery, and 1.5% in elective surgery (p < 0.0002), and 4.5% in CABG alone and 7.4% in CABG with concomitant surgery (p = 0.3763), and 25% in patients receiving vein grafts only and 3.0% in those receiving at least 1 artery graft (p = 0.0003). Overall patient distribution was 32% good, 20% fair, 20% poor, 11% high-risk, and 16% extremely high-risk. Predicted mortality was 2.2% for patients who were a good risk, 6.7% for fair-risk, 12% for poor-risk, 16% for high-risk, and 25% for extremely high-risk patients. Actual operative mortality was 1.0% for good-risk, 0% for fair-risk, 3.4% for poor-risk, 6.3% for high-risk, and 18% for extremely high-risk patients, making actual mortality significantly lower than that predicted. CONCLUSION: Comparing predicted mortality and actual mortality enabled us to objectively calculate operative results and assess operative quality.

Observation of everyday hand-washing behavior of Japanese, and effects of antibacterial soap.

People wash their hands only for a short time outside the home and when preparing meals at home. This may not be sufficient for those who prepare meals because of possible secondary contamination from food. Although washing with a placebo soap for a short period (lathering 3 s and rinsing 8 s) cleansed from hands about 95% of the total coliforms transferred from ground meat, an antibacterial soap further reduced the coliform count significantly (p < 0.01). To effectively avoid secondary contamination, it is recommended that people should more frequently wash their hands, using an antibacterial soap on the areas that have been in contact with raw meat, poultry, seafood, eggs, vegetables and other foods.

[Early and mid-term results after cardiac or thoracic aortic surgery in patients aged 70 years and older--survival and quality of life].

The early and mid-term survival after cardiac or thoracic aortic surgery and influence of age to the operative mortality were examined in 168 consecutive patients aged 70 years and older from August 1994 to December 1998, together with assessment of postoperative quality of life (QOL). The mean age was 74.1 +/- 3.7 years old (70 to 86 years). 80 patients had IHD, 59 had VHD, 28 had TAA. Preoperative risk score was classified in 5 groups (good, fair, poor, high, extremely high) based on Parsonnet method. Current QOL of the survivors was assessed using Asanoi method with questionnaire by a letter. There were 9 early deaths (5.4%). Following the Parsonnet model observed mortality was absolutely lower than predicted mortality. When age score was excluded in the Parsonnet model, the observed mortality became almost equal with the predicted mortality. There were 22 late deaths (6.9%/P-Y). The actuarial survival rate of age 70 to 74 group was significantly higher than the age 75 years and older group (p = 0.0021). The actuarial survival rate of TAA group was significantly lower than the VHD or IHD group (p < 0.02). Postoperative NYHA and activity score of TAA group were better than VHD or IHD group. We got satisfactory answers for operation in 95% current survivors. Patients aged 70 years and older will be undergone cardiac or thoracic aortic surgery at a reasonable risk and well satisfaction.

A new model of cerebral microthrombosis in rats and the neuroprotective effect of a Rho-kinase inhibitor.

BACKGROUND AND PURPOSE: The aim of this study was to develop a new model of stroke based on endothelial damage and thrombotic occlusion in a perforating artery, leading to small cerebral infarcts and neurological deficits in rats. Moreover, the neuroprotective efficacy of fasudil, a rho-kinase inhibitor, was investigated in this model. METHODS: Fifty-six male Sprague-Dawley rats were used in the present study. Rats were anesthetized with sodium pentobarbital, and 100 microg of sodium laurate was injected into the left internal carotid artery on days 1 and 3. The thrombus induction and consequent of ischemic brain damage were examined by histopathological analyses and neurological deficit scoring in a posture reflex test. To investigate the neuroprotective effects of fasudil, 1 or 10 mg/kg was administered intraperitoneally 5 minutes after the first injection of sodium laurate and once daily thereafter on the following 2 days. RESULTS: One hour after the injection of sodium laurate, microscopic examination of phosphotungstic acid hematoxylin-stained sections (n=5) revealed that microthrombi containing fibrin strands obstructed the perforating arteries in the ipsilateral hemisphere. Under a transmission electron microscope (n=6), endothelial cells appeared exfoliated and the vascular lumen was obstructed by a thrombus composed of degranulated platelets, fibrin, leukocytes, and erythrocytes. No evidence of endothelial cell damage or thrombus could be found in the ipsilateral side of the pial artery (middle cerebral artery). Twenty-four hours after the second injection of sodium laurate (day 4), 13 of 15 rats (86.6%) showed mild to severe neurological deficits. Multiple small cerebral infarcts were observed in the hippocampus, cortex, and thalamus. Treatment with fasudil (1 and 10 mg/kg, n=15 each) resulted in a significant improvement in neurological deficits. Fasudil also significantly reduced the area of cerebral infarction. CONCLUSIONS: We present a new model of stroke in rats, in which the perforating arteries are selectively occluded by microthrombi. This model is useful to investigate the pathophysiology and treatment of small cerebral infarction, which is caused by perforating arterial occlusive diseases such as lacunar infarcts. Fasudil may be beneficial in the treatment of acute ischemic stroke.

Formation and characterization of planar lipid bilayer membranes from synthetic phytanyl-chained glycolipids.

The formability, current-voltage characteristics and stability of the planar lipid bilayer membranes from the synthetic phytanyl-chained glycolipids, 1, 3-di-O-phytanyl-2-O-(beta-glycosyl)glycerols (Glc(Phyt)(2), Mal(N)(Phyt)(2)) were studied. The single bilayer membranes were successfully formed from the glycolipid bearing a maltotriosyl group (Mal(3)(Phyt)(2)) by the folding method among the synthetic glycolipids examined. The membrane conductance of Mal(3)(Phyt)(2) bilayers in 100 mM KCl solution was significantly lower than that of natural phospholipid, soybean phospholipids (SBPL) bilayers, and comparable to that of 1,2-diphytanoyl-sn-glycero-3-phosphocholine (DPhPC) bilayers. From the permeation measurements of lipophilic ions through Mal(3)(Phyt)(2) and DPhPC bilayers, it could be presumed that the carbonyl groups in glycerol backbone of the lipid molecule are not necessarily required for the total dipole potential barrier against cations in Mal(3)(Phyt)(2) bilayer. The stability of Mal(3)(Phyt)(2) bilayers against long-term standing and external electric field change was rather high, compared with SBPL bilayers. Furthermore, a preliminary experiment over the functional incorporation of membrane proteins was demonstrated employing the channel proteins derived from octopus retina microvilli vesicles. The channel proteins were functionally incorporated into Mal(3)(Phyt)(2) bilayers in the presence of a negatively charged glycolipid. From these observations, synthetic phytanyl-chained glycolipid bilayers are promising materials for reconstitution and transport studies of membrane proteins.

Acute type A aortic dissection involving the left main trunk of the coronary artery--a report of two successful cases.

This report describes 2 cases of a type A acute aortic dissection combined with myocardial infarction caused by a retrograde dissection into the left main trunk of the coronary artery. Successful surgical treatments, including the replacement of the ascending aorta, aortic valve resuspension and coronary artery bypass grafting, were performed in both patients, and they recovered well from cardiogenic shock. However, left ventricular function of both patients remained depressed postoperatively, which limited their quality of life. Because no definite method for salvaging infarcted myocardium has yet been established, either more timely surgery or the preoperative placement of a perfusion catheter in the left main coronary artery is mandatory.

Fasudil, a protein kinase inhibitor, prevents the development of endothelial injury and neutrophil infiltration in a two-haemorrhage canine subarachnoid model.

We examined the possible prophylactic potential of fasudil, a protein kinase inhibitor, on the development of endothelial injury and neutrophil infiltration after subarachnoid haemorrhage (SAH). Using the two haemorrhage canine model, fasudil (3 mg/kg) was infused intravenously for 30 min twice daily (days 1-7) and related histological changes were observed by light and electron microscopy. On day 7 characteristic features of the basilar arteries included corrugation of the elastic lamina and endothelial disruption; fasudil inhibited this endothelial damage. Marked neutrophil infiltration into the subarachnoid space was not detected until day 3. On day 7 a large number of neutrophils was observed in the subarachnoid space around all the basilar arteries examined; fasudil treatment significantly inhibited neutrophil infiltration. Our findings suggest that: (1) endothelial injury and neutrophils play a major role in the pathogenesis of cerebral vasospasm; and (2) fasudil inhibited both endothelial damage and neutrophil infiltration, and therefore protein kinase pathways may have a role in these pathological events. Copyright 1999 Harcourt Publishers Ltd.

Thromboxane A2 synthetase inhibitor failed to ameliorate the arterial narrowing during the chronic phase of cerebral vasospasm.

To determine the pathogenetic mechanism underlying the maintenance of arterial narrowing during the chronic phase of cerebral vasospasm caused by subarachnoid hemorrhage (SAH), we examined the effect of ozagrel, a thromboxane A2 (TXA2) synthetase inhibitor, on chronic vasospasm in a canine two-hemorrhage model in comparison with that of fasudil, an inhibitor of protein kinases. The magnitude of the vasospasm was determined angiographically. On SAH day 7, a vasospasm was observed in every dog. Intraarterial or intravenous administration of ozagrel (3 mg/kg/30 min) did not reverse the vasospasm but tended to increase bleeding. In contrast, intraarterial administration of fasudil (3 mg/kg/30 min) significantly reversed the vasospasm. These findings suggest that: 1) TXA2 does not participate in the maintenance of chronic vasospasm after SAH; and 2) the protein kinases, particularly myosin-light chain kinase and protein kinase C, are involved in the pathogenesis of arterial narrowing during the chronic phase of cerebral vasospasm.

[Questionnaire of cryopreserved allograft valves in Kyushu and Yamaguchi area].

Japanese Society of Cryopreserved Thoracic Tissue Implantation was recently set up. Cryopreserved allograft valves are about to pervade in Japan. To clarify the interest, demands and supply of cryopreserved allograft valve in the area of Kyushu and Yamaguchi, we performed questionnaire investigation regarding this issue. Collection rate of this questionnaire was 87.5% (35/40 hospitals). Ninety-seven percent of the hospitals answered that they were interested in cryopreserved allograft. Ninety-four percent of the hospitals answered that they did not ethically hesitate to use cryopreserved allograft. Ninety-one percent of the hospitals hoped to perform cardiac surgery using allograft aortic valve if allograft is available. With respect to securing donors of allograft, two-third of the hospitals did not decide whether they could be involved in obtaining donors of allograft. As the number of the cadaver kidney donors is about 20 per year in the area of Kyushu and Yamaguchi, shortage of donors of allograft valve is anticipated.

Postoperative cardiac rhythms with superior-septal approach and lateral approach to the mitral valve.

BACKGROUND: The superior-septal approach provides an excellent view of the mitral valve and therefore has received considerable interest. However, the safety of this approach is controversial because it requires division of the sinus node artery in most cases. METHODS: Postoperative cardiac rhythms were analyzed in 152 consecutive patients who underwent mitral valve procedures between January 1992 and February 1995 with a conventional right lateral left atriotomy (group 1, n = 69) or the superior-septal approach (group 2, n = 83). Follow-up ranged from 2 to 38 months, and the mean follow-up was 16.1 months in group 1 and 13.8 months in group 2. RESULTS: The mortality rate was similar in the two groups (1.4% in group 1 and 1.2% in group 2), and the causes of death were not related to the left atriotomy. At discharge, 96% of the patients in group 1 who were in sinus rhythm preoperatively and 78% of those in group 2 remained in sinus rhythm. At the last follow-up, 88% of these patients in group 1 and 83% in group 2 remained in sinus rhythm. Among the patients in atrial fibrillation or junctional rhythm before operation, 12% in group 1 and 11% in group 2 had regained sinus rhythm at the last follow-up. There were no significant differences in these values. CONCLUSIONS: Although the incidence of dysrhythmias was higher with the superior-septal approach in the early postoperative period, this approach provides an excellent operative view of the mitral valve and similar results in terms of late postoperative cardiac rhythms as the right lateral left atriotomy.

Development of the Bowman's and Jacobson's glands in the Japanese reddish frog, Rana japonica.

The olfactory organs are equipped with their own associated glands, Bowman's glands (BG) of the olfactory epithelium (OE), and Jacobson's glands (JG) of the vomeronasal organ (VNO). Histology and ultrastructure of these glands in the adult were investigated in the Japanese reddish frog, Rana japonica, along with their development from hatching to the end of metamorphosis. In the adult, BG cells contained large, electron-opaque secretory granules, intensely PAS-positive by light microscopy, while JG cells contained middle-sized secretory granules with moderate electron density, only faintly PAS-positive. Embryologically, BG appeared within the OE at 44 days after hatch, increased in number and were situated in the lamina propria under the OE at 52 days after hatch. BG cells contained large, electron-opaque granules, and well-developed rER at this time. While, JG appeared much earlier than BG. The VNO appeared as a concave of the ventral part of the OE at 4 days after hatch, and JG appeared under the VNO at 10 days after hatch. JG cells contained well-developed rER at 12 days after hatch. Secretory granules appeared in a small number in JG at 24 days after hatch, and increased thereafter. These findings suggest that JG may take part in secretion earlier than BG in ontogeny.

Development of the olfactory epithelium and vomeronasal organ in the Japanese reddish frog, Rana japonica.

Histological and ultrastructural development of the olfactory epithelium (OE) and vomeronasal organ (VNO) was investigated in the Japanese reddish frog, Rana japonica. Tadpoles, from hatching to the end of metamorphosis, and adult frogs were examined. In the adult, olfactory cells of the OE were equipped with olfactory vesicles with long cilia, but supporting cells with microvilli. The supporting cells of the OE contained secretory granules, PAS-positive by light microscopy, in their apical cytoplasm. On the contrary, sensory cells of the VNO were equipped with microvilli, and supporting cells of the VNO were equipped with cilia, but without secretory granules. Embryologically, the olfactory cells were indistinguishable from the supporting cells in the olfactory placode (primitive OE) lining the nasal pit, at hatch. The VNO appeared as a concave of the ventral part of the OE at 4 days after hatch. At the time, the olfactory and supporting cells of the OE became distinguishable from each other. Secretory granules were formed in the supporting cells of the OE at 36 days after hatch, and the OE was similar in fine structure to that in the adult. While, the VNO remained immature at 24-36 days after hatch, and did not complete its ultrastructural development at 60 days after hatch, the end of metamorphosis. These findings suggest that the OE may take part in the olfaction earlier than the VNO in ontogeny.

Simplified Manouguian's aortic annular enlargement for aortic valve replacement.

A simplified Manouguian's aortic annular enlargement for aortic valve replacement with prosthetic valve was performed in a patient with severe aortic stenosis and mild regurgitation by preserving both the anterior mitral leaflet and the left atrial roof intact. This method minimized the operative risk by avoiding injury to the mitral valve, while inserting a possibly two-size-larger prosthetic valve. The inserted prosthetic valve was positioned parallel to the original aortic valve. Moreover, the incision from the top of the commissure between the noncoronary cusp and left coronary cusp through the interventricular fibrous trigone appears to be easily extended into the exact center of the anterior mitral leaflet if further enlargement is required.