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BACKGROUND: Meticillin-resistant Staphylococcus aureus (MRSA) is the most common pathogen in orthopaedic surgical site infections (SSIs). However, few studies have investigated the transmission process of orthopaedic MRSA SSI. AIM: To investigate the transmission process of orthopaedic MRSA SSI using epidemiological and molecular analyses and to determine a method to prevent MRSA SSI in nosocomial orthopaedic surgery. METHODS: Active MRSA surveillance, preoperative decolonization and contact precautions for MRSA-positive cases was performed at our institution. Changes in epidemic strains were evaluated and the possibility of transmission from patients in an orthopaedic ward of a Japanese tertiary-care hospital was assessed by genotyping stored MRSA strains. In addition, data on the prevalence of MRSA SSI, MRSA colonization, and use of an alcohol antiseptic agent (mL/patient-days) during 2005-2022 were retrospectively assessed. FINDINGS: SCCmec type II strain in the SSI group decreased over time, associated with fewer outbreaks. Even during a period of high infection rates, no cases of transmission-induced SSI from nasal MRSA carriers were identified. The infection rate correlated negatively with the use of an alcohol antiseptic agent (r = -0.82; P < 0.0001). Two cases among five nasal carriers developed MRSA SSI caused by strains different from those related to nasal colonization. CONCLUSION: The infection control measures for transmission from the hospital reservoirs including strict adherence to hand hygiene and decolonization of carriers is likely to be important for the prevention of orthopaedic MRSA SSI. However, the need for contact precautions for decolonized nasal carriers might be low.

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BACKGROUND: The measurement of waist circumference (WC) is the most prevalent cause of the metabolic syndrome (MS). OBJECTIVE: The aim of this study was to correlate WC and BMI with high-density lipoprotein (HDL-c) levels in patients with MS being consulted by the Family Health Program (PSF), Brazil. METHODS: This cross-sectional study was conducted from September to November 2008 with 42 patients (29 women and 13 men) from 35 to 77 years. Dietary intake was reported, and biochemical and body composition measures were taken. RESULTS: The HDL-c levels were higher in women when compared to men (48.4 ± 8.1 mg/dL vs. 36.4 ± 7.8 mg/dL). However, the triglycerides (TG)/HDL-c ratio and TG concentrations were lower in women (3.8 ± 1.5 and 178.0 ± 57.8 mg/dL, respectively) than in men (9.4 ± 8.5 and 471.5 ± 501.5 mg/dL, respectively). Regarding skinfold profile, the triceps was greater in females (37.0 ± 8.4 cm vs. 20.7 ± 10.5 cm). The dietetic profile showed that women had a lower intake of energy, fiber, phosphorus and sodium. The fruits and vegetables intake was diminished in the participants of this study, as less than 60% of the women and 50% of men met the daily recommendations. Approximately 54% of men and 28% of women had a lower intake of dairy products daily. Moreover, the results shows that the WC was negatively correlated to HDL-c (r = -0.41, p < 0.05) whereas the BMI is not associated with HDL-c (r = -0.34, p > 0.06). CONCLUSION: Our findings showed that WC is a better predictor of changes in HDL-c than BMI.

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BACKGROUND: Oral clonidine, an alpha2-adrenergic receptor agonist, reduces the dose of propofol required for laryngeal mask airway (LMA) insertion. Target-controlled infusion (TCI) is becoming increasingly popular for propofol infusion. There is no information, however, on the propofol blood concentrations required for LMA insertion and the effect of oral clonidine premedication on these values. METHODS: Propofol at target effect-site concentrations from 4.0 to 12.0 microg/ml were randomly administered using TCI in three groups of healthy male patients (n=35 each) who were undergoing elective orthopedic surgery: control, 2.5 microg/kg clonidine, and 5.0 microg/kg clonidine groups. Nothing was administered to the control group. Clonidine(2.5 microg/kg or 5.0 microg/kg) was administered orally 90 min before arrival at the operating room in the clonidine groups. After equilibration between the blood- and effect-site for 15 min, insertion of the LMA was attempted. The EC50 for LMA insertion (measured propofol serum concentration in equilibrium with the effect-site at which 50% of patients do not respond to the insertion of the LMA) was determined by logistical regression. RESULTS: EC50+/-standard error values in the control, 2.5 microg/kg clonidine, and 5.0 microg/kg clonidine groups were 8.72+/-0.55, 7.76+/-0.60, and 5.84+/-0.58 microg/ml, respectively. The EC50 in the 5.0 microg/kg clonidine group was significantly lower than that in the control group (P < 0.01). CONCLUSIONS: The propofol concentration required for LMA insertion in healthy male patients is reduced by premedication with 5.0 microg/kg oral clonidine.

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UNLABELLED: Requirements for inhaled anesthetics decrease during pregnancy. There are no published data, however, regarding propofol requirements in these patients. Because propofol is often used for induction of general anesthesia when surgery is necessary in early pregnancy, we investigated whether early pregnancy reduces the requirement of propofol for loss of consciousness using a computer-assisted target-controlled infusion (TCI). Propofol was administered using TCI to provide stable concentrations and to allow equilibration between blood and effect-site (central compartment) concentrations. Randomly selected target concentrations of propofol (1.5-4.5 microg/mL) were administered to both pregnant women (n = 36) who were scheduled for pregnancy termination and nonpregnant women (n = 36) who were scheduled for elective orthopedic or otorhinolaryngologic surgery. The median gestation of the pregnant women was 8 wk (range, 6-12 wk). Venous blood samples for analysis of the serum propofol concentration were taken at 3 min and 8 min after equilibration of the propofol concentration. After a 10-min equilibration period of the predetermined propofol blood concentration, a verbal command to open their eyes was given to the patients twice, accompanied by rubbing of their shoulders. Serum propofol concentrations at which 50% of the patients did not respond to verbal commands (C(50) for loss of consciousness) were determined by logistic regression. There was no significant difference in C(50) +/- SE of propofol for loss of consciousness between the Nonpregnant (2.1 +/- 0.2 microg/mL) and Pregnant (2.0 +/- 0.2 microg/mL) groups. These results indicate that early pregnancy does not decrease the concentration of propofol required for loss of consciousness. IMPLICATIONS: The C(50) of propofol for loss of consciousness in early pregnancy did not differ from that in nonpregnant women, indicating that there is no need to decrease the propofol concentration for loss of consciousness when inducing general anesthesia for termination of pregnancy.

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UNLABELLED: A new CO(2) absorbent, Amsorb (A), which does not contain monovalent bases, is ideal because it does not degrade volatile anesthetics to either Compound A (from sevoflurane) or carbon monoxide (from desflurane, enflurane, or isoflurane). The CO(2) absorption capacity of A, however, has not been investigated under clinical conditions. In this study, we compared the longevity (time to exhaustion) and CO(2) absorption capacity (the volume of CO(2) absorbed before CO(2) rebreathing occurs) of A under low-flow anesthesia (1 L/min) with those of two soda lime absorbents-Medisorb (M) and Sodasorb (S)-by using a 750-mL ADU canister and a 1350-mL Aestiva 3000 canister. In the study with the ADU canister, the longevity of A was 213 +/- 71 min, significantly less than those of M (445 +/- 125; P < 0.01) and S (503 +/- 89; P < 0.001). The CO(2) absorption capacity (L/100 g absorbent) of A was 5.5 +/- 1.2, significantly less than those of M (10.7 +/- 1.7) and S (12.1 +/- 1.8; P < 0.001). In the study with the Aestiva 3000 canister, the longevity of A was 218 +/- 61 min, significantly less than those of M (538 +/- 136) and S (528 +/- 103; P < 0.001). The CO(2) absorption capacity (L/100 g absorbent) of A was 7.6 +/- 1.6, significantly less than those of M (14.4 +/- 1.8) and S (14.8 +/- 2.3; P < 0.001). These results indicate that the CO(2) absorption capacity of A is half that of M or S and that the difference in the CO(2) absorption capacity between A and M or S is almost constant, regardless of the canister design. IMPLICATIONS: The CO(2) absorption capacity of Amsorb is half that of Medisorb and Sodasorb under clinical low-flow (1 L/min) anesthesia with either a 750-mL Ohmeda ADU compact or a 1350-mL Ohmeda Aestiva 3000 canister.

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Tubular projections from plastids (stromules) were observed using a stroma-targeted green fluorescent protein (GFP) fusion protein and a confocal laser scanning microscope. In tobacco (Nicotiana tabacum) epidermal cells, stromules were observed at a high frequency. Some of them were long and connected plastids. Three days after particle bombardment, 80.6+/-6.99% of the transformed cells contained some plastids with more than one stromule, and 40.2+/-7.7% contained at least one pair of plastids connected by stromules. In a few cells, numerous and highly developed stromules covering the whole cell were observed. Stromules were also observed in epidermal cells in each of three other plant species that were tested: rice, dayflower (Commelina communis) and Arabidopsis thaliana. These findings demonstrated that stromules are common structure in plant epidermal cells.

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It is known that alcoholic fermentation is important for survival of plants under anaerobic conditions. Acetaldehyde, one of the intermediates of alcoholic fermentation, is not only reduced by alcohol dehydrogenase but also can be oxidized by aldehyde dehydrogenase (ALDH). To determine whether ALDH plays a role in anaerobic metabolism in rice (Oryza sativa L. cv Nipponbare), we characterized a cDNA clone encoding mitochondrial ALDH from rice (Aldh2a). Analysis of sub-cellular localization of ALDH2a protein using green fluorescent protein and an in vitro ALDH assay using protein extracts from Escherichia coli cells that overexpressed ALDH2a indicated that ALDH2a functions in the oxidation of acetaldehyde in mitochondria. A Southern-blot analysis indicated that mitochondrial ALDH is encoded by at least two genes in rice. We found that the Aldh2a mRNA was present at high levels in leaves of dark-grown seedlings, mature leaf sheaths, and panicles. It is interesting that expression of the rice Aldh2a gene, unlike the expression of the tobacco (Nicotiana tabacum) Aldh2a gene, was induced in rice seedlings by submergence. Experiments with ruthenium red, which is a blocker of Ca(2+) fluxes in rice as well as maize (Zea mays), suggest that the induction of expression of Adh1 and Pdc1 by low oxygen stress is regulated by elevation of the cytosolic Ca(2+) level. However, the induction of Aldh2a gene expression may not be controlled by the cytosolic Ca(2+) level elevation. A possible involvement of ALDH2a in the submergence tolerance of rice is discussed.

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UNLABELLED: Sevoflurane degrades to Compound A, which is nephrotoxic in rats. Potassium hydroxide (KOH) and sodium hydroxide (NaOH) are primary determinants of this degradation reaction. To address this, new carbon dioxide absorbents, such as Amsorb((R)) (A; Armstrong Medical, Coleraine, Northern Ireland), which contains neither KOH nor NaOH, Drägersorb 800 Plus((R)) (D; Dräger, Luebeck, Germany), and Medisorb((R)) (M; Datex-Ohmeda, Bromma, Sweden), which contain some NaOH (1% to 2%) and only trace amounts of KOH (0.003%), were recently developed. We compared Compound A concentrations using these three CO(2) absorbents during low-flow (1 L/min) sevoflurane anesthesia in surgical patients, with those using a conventional CO(2) absorbent, Drägersorb 800 (C). The mean Compound A concentrations +/- SD using C, A, D, and M were 18.7 +/- 2.5, 1.8 +/- 0.7, 13.3 +/- 3.5, and 11.2 +/- 2.6 ppm, respectively, with significant differences (P < 0.001; A versus C, A versus D, A versus M, C versus D, C versus M). Amsorb prevented the degradation of sevoflurane to Compound A, whereas Drägersorb 800 Plus and Medisorb decreased the degradation to Compound A. IMPLICATIONS: Sevoflurane degradation to Compound A is decreased by lowering the concentration of monovalent bases in the carbon dioxide absorbent (Drägersorb 800 Plus) [Dräger, Luebeck, Germany] and Medisorb) [Datex-Ohmeda, Bromma, Sweden]) and is virtually eliminated in the absence of these bases (Amsorb) [Armstrong Medical, Coleraine, Northern Ireland]).

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We have developed a new device to measure the friction force and calculate the friction coefficient between a rabbit flexor tendon, a pulley and a proximal phalanx. The flexor digitorum fibularis tendon of a rabbit was taken intact with the proximal phalanx, and tendon pulleys were attached to both ends of the bone. Both ends of the tendon were clamped to acrylic plates and connected to stainless-steel plates equipped with strain gauges. A pretension of 1.96 N was applied so as not to loosen the tendon. The proximal phalanx was fixed to an acrylic plate on the actuator, which gave 8 mm of transfer to the acrylic plate at a speed of 2 mm/s. The interface between the tendon and the surrounded tissue created the friction force, when the load was applied on the distal pulley. The friction force could be obtained from the difference between the tension of both ends of the tendon, which was measured with strain gauges and sampled with a personal computer. The friction force and the friction coefficient were calculated from the measured force and the applied load. The load and the pre-loading time, which was defined as loading duration before gliding, were varied in order to observe the change of the friction coefficient. The friction coefficient was not affected by the load and increased with the pre-loading time. The value of mu(s) ranged from 0.027 to 0.111 (0.072 +/- 0.023), and that of (mu)d ranged from 0.010 to 0.069 (0.039 +/- 0.014) (pre-loading time was 5 s). Our method will allow for the examination of various surgical treatments and lubricants. Moreover, it can be applied to other tissues of any animals with similar structures to the rabbit's digitorum.

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We have cloned a novel nuclear gene for a ribosomal protein of rice and Arabidopsis that is like the bacterial ribosomal protein S9. To determine the subcellular localization of the gene product, we fused the N-terminal region and green fluorescent protein and expressed it transiently in rice seedlings. Localized fluorescence was detectable only in chloroplasts, indicating that this nuclear gene encodes chloroplast ribosomal protein S9. The N-terminal region of rice ribosomal protein S9 was found to have a high sequence similarity to the transit peptide region of the rice chloroplast ribosomal protein L12, suggesting that these transit peptides have a common lineage.

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A tyrosine kinase receptor-mediated and a heterotrimeric G protein-coupled receptor-mediated signals have been shown to evoke distinct intracellular signaling events. There has been increasing evidence that cross-talk exists between a tyrosine kinase receptor-mediated and a heterotrimeric G protein-coupled receptor-mediated signal transduction pathways. In the present study, we have studied effects of EGF receptor activation on activities of inhibitory G protein (Gi). We show that the amounts of Gi/Go ADP-ribosylated by islet-activating protein (IAP) increased by 30-40% in the membranes of Rat 1 fibroblast cells pretreated with EGF compared with those without pretreatment. When an effect of lysophosphatidic acid (LPA) stimulation on an adenylate cyclase activity was examined, LPA partly attenuated forskolin-stimulated adenylate cyclase activity via Gi because IAP pretreatment blocked the inhibitory effect of LPA. Pretreatment with EGF reduced the ability of LPA to inhibit the forskolin-stimulated adenylate cyclase activity, while the pretreatment did not have any effects on the forskolin-stimulated activity. Thus, the EGF receptor-mediated signal appears to cause the impairment of Gi function in Rat 1 fibroblast cells.

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GTPase-activating proteins (GAPs) stimulate the hydrolysis of GTP bound to small G-proteins and regulate the signal transduction pathway. Changes in the expression of p21-Ras p120-GAP induced by growth factor treatment were examined in cultured Chinese hamster ovary (CHO) and human choriocarcinoma (BeWo) cells. Expression of p120-GAP and GAP activity were measured. Fetal bovine serum induced a significant increased level of GAP in CHO cells, but did not increase GAP in BeWo cells. The results suggest that growth factors affect Ras GAP expression in CHO cells, while they do not in other cells such as BeWo cells.

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Ras GTPase-activating protein (GAP), an important downregulator of Ras activity, has previously been shown to be abundant in human placenta. The expression of p120 and p100 isoforms of GAP in human normal chorionic villi (n=5) and hydatidiform mole (n=5) was investigated to clarify the involvement of Ras GAP in the growth of chorionic villi in the first trimester of pregnancy. Immunoblot analysis revealed that both p120- and p100-GAP isoforms were remarkably less expressed in mole villi than in normal chorionic villi. The expression of p100-GAP significantly reduced in comparison with that of pl20-GAP in mole villi. Northern blot analysis showed that the amount of GAP mRNA reduced in hydatidiform mole less than one-third of that in normal chorionic villi. The GAP activity, measured by the effect of tissue extract on the hydrolysis of Ras-bound GTP, was significantly lower in hydatidiform mole than in normal chorionic villi. These results suggest that Ras GAP may play an important role in the normal growth and differentiation of human chorionic villi in the first trimester.

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Protein phosphorylation is one of several representative post-translational modifications. Cyclic AMP-dependent protein kinase (PKA) plays the crucial and varying role of signal transduction. On the other hand, ras proteins plays an important role in cell proliferation and growth. Although a previous report showed that H-ras protein was phosphorylated by PKA, the stoichiometry was not determined, so we investigated the stoichiometry of phosphorylation of the protein by PKA. H-ras cDNA inserted into a pGEX-2T expressing vector produced high levels of recombinant H-ras (rH-ras) in a fusion protein with glutathione S-transferase. rH-ras was obtained after cleavage by thrombin. Phosphorylation of ras protein by the catalytic subunit of PKA was performed, and the radioactivity was counted after SDS-PAGE and autoradiography. The results indicate that less than 0.1 mol of phosphate was incorporated per mol of H-ras protein, and suggest that H-ras protein could not be a physiologically meaningful substrate for PKA.

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A case of a 53-year-old man with left pheochromocytoma associated with chronic renal failure is described. He had been on regular hemodialysis for 15 years. Also, because of right renal carcinoma, he underwent right nephrectomy at the same time. For preoperative preparation, we kept the patient's dry weight. Anesthesia was induced with isoflurane 1.0% in oxygen, and fentanyl 0.3 mg and midazolam 5 mg, followed by tracheal intubation facilitated with vecuronium. Anesthesia was maintained with isoflurane 0.5% in oxygen 50-100%, fentanyl and midazolam. Intraoperative hypertension during the manipulation of the tumor and hypotension after removal of the tumor were controlled by continuous or bolus infusion of phentolamine and norepinephrine respectively. For intraoperative fluid management, we administered crystalloid solution 4 ml.kg-1.hr-1. Careful preoperative management was carried out to prevent severe intraoperative and postoperative cardiovascular or respiratory complications.

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In the multifunctional pathogens of atherosclerosis, oxidatively modified low density lipoprotein (LDL) plays a central role in atherogenesis. We searched to find out whether oxidized LDL (ox-LDL) could induce apoptosis in smooth muscle cells (SMCs). The induction of apoptosis was demonstrated by terminal deoxynucleotidyl transferase mediated dUTP-biotin nick end labeling and DNA fragmentation. Ox-LDL induced apoptosis in a concentration dependent manner in the cells. The structural and biological changes in ox-LDL may be attributed to lysophosphatidylcholine (lyso-PC) accumulation and lipid peroxidation. To determine whether lyso-PC or lipid peroxide is responsible for the biological effect of ox-LDL, we incubated SMCs with lyso-PC or 7-ketocholesterol. Lyso-PC did not induce apoptosis, but 7-ketocholesterol did induce apoptosis. We conclude that ox-LDL induces apoptosis in SMCs and that this apoptosis contributes to lipid peroxidation. This mechanism may be important in determining the course of atherogenesis.

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In the present study, we investigated the mechanism of phenylephrine (alpha-1-adrenergic receptor agonist)-induced arachidonic acid release in Japanese white rabbit aortic smooth muscle cells (SMC). When introduced into permeabilized smooth muscle cells, guanosine S-[gamma-thio] triphosphate (GTPgamma S), which activates GTP-binding proteins (G proteins), stimulated arachidonic acid (AA) release. Neomycin, an inhibitor of phosphoinositide (PI) turnover, was almost without effect on GTP[gamma S] stimulated AA release. In addition, pertussis toxin (PT) partially inhibited phenylephrine-stimulated AA release, suggesting that IAP (Islet activating protein)-sensitive G proteins mediate this process. Phenylephrine-stimulated AA release was also inhibited by decreased extracellular calcium and aristolochic acid, suggesting a role for a phospholipase A2 (PLA2). Next PLA2 is reported to be a substrate for mitogen-activated protein (MAP) kinase. We examined the effect of phenylephrine on MAP kinase and c-jun N-terminal kinase (JNK) phosphorylation. Phenylephrine didn't induce phosphorylation of MAP kinase, but did induce phosphorylation of JNK. In addition, cells which were pretreated with PT inhibited the phosphorylation of JNK. These results suggest that IAP-sensitive G protein is involved in the coupling between alpha1-adrenergic receptor (AR) and PLA2 in cultured rabbit aortic SMCs, and that the alpha1-AR-induced AA release is mediated by JNK.

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BACKGROUND: In studies of methoxyflurane-induced nephrotoxicity, renal-concentrating impairment has been observed only when serum inorganic fluoride concentrations exceed 50 microM. Prolonged sevoflurane anesthesia can result in serum inorganic fluoride concentrations in excess of 50 microM. The authors compared renal function after prolonged sevoflurane anesthesia with that after isoflurane anesthesia. In addition, they measured urinary excretion of N-acetyl-beta-glucosaminidase (NAG), a sensitive index of renal tubular damage, during the 3-day period after anesthesia. METHODS: Thirty-four healthy patients who underwent either sevoflurane (23 patients) or isoflurane (11 patients) anesthesia at a total gas flow of 61/min for orthopedic surgery scheduled to last at least 5 h were studied. At 16.5 h after cessation of anesthesia, patients were administered 10 units of vasopressin and urine was collected frequently thereafter for evaluation of urinary osmolality. In addition, urinary excretion of NAG was measured before and on days 1-3 after anesthesia. Based on whether peak fluoride concentrations exceeded 50 microM, 23 patients anesthetized with sevoflurane were assigned to a sevofluranehigh group (> 50 microM) or a sevofluranelow (< 50 microM) group. RESULTS: The eight patients in the sevofluranehigh group had a mean peak fluoride concentration of 57.5 +/- 4.3 microM. A significant, albeit weak, inverse correlation was found between peak fluoride concentration and maximal urinary osmolality after the injection of vasopressin (r = -0.42, P < 0.05). Mean maximum urinary osmolality tended to be lower in the sevofluranehigh group (681 +/- 60 mOsm/kg) than in the other two groups after administration of vasopressin, although the difference among the three groups did not quite reach a statistical significance (P = 0.068). One patient had a transient concentrating defect (maximum urinary osmolality = 390 mOsm/kg) on day 1 after anesthesia. Urinary excretion of NAG in both the sevofluranehigh and sevofluranelow groups was greater on days 2 and 3 after anesthesia than before anesthesia. The increase in urinary NAG excretion was dose related with sevoflurane, but there was no difference in results of routine laboratory renal tests on days 2 and 3 after anesthesia among the three groups. CONCLUSIONS: The authors concluded that sevoflurane anesthesia results in increased serum fluoride concentration, a tendency toward decreased maximal ability to concentrate urine, and increased excretion of NAG. However, the increase in urinary NAG excretion was not indicative of clinically significant renal damage in these patients with no preexisting renal disease.

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The effect of wortmannin, which inhibits phosphatidylinositol 3-kinase (PI 3-kinase), on adipocytic differentiation of 3T3-L1 cells was examined. The extent of differentiation was evaluated by the staining of adipocytes with Oil Red O and measurement of glycerol 3-phosphate dehydrogenase (GPDH) activity. Wortmannin, at over 100 nM, significantly inhibited adipogenesis of cells treated with isobutyl-methylxanthine, dexamethasone, and insulin with an IC50 value about 50 nM. These results suggest that PI 3-kinase plays a role in adipocytic differentiation of 3T3-L1 cells.

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Few studies have used the vasopressin test to evaluate urine concentrating ability after sevoflurane anaesthesia. We performed a vasopressin test on the first day after operation to compare the effect of prolonged sevoflurane anaesthesia for orthopaedic procedures (n = 11) with that of isoflurane (n = 10). Mean doses of sevoflurane and isoflurane were 10.6 (SE 0.9) and 8.5 (1.5) MAC-h, respectively. Mean peak serum fluoride concentration in patients anaesthetized with sevoflurane was 41.9 (2.5 mumol litre-1 and exceeded 20 mumol litre-1 for approximately 20 h. Each group showed similar responses to vasopressin. There was no evidence of subclinical nephrotoxicity in patients given prolonged sevoflurane anaesthesia.

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