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Victims of fire accidents who sustain both thermal injury to the skin and smoke inhalation have gross evidence of oxidant injury. Therefore, we hypothesized that delivery of vitamin E, an oxygen superoxide scavenger, directly into the airway would attenuate acute lung injury postburn and smoke inhalation. Sheep (N = 17 female, 35 +/- 5 kg) were divided into 3 groups: (1) injured, then nebulized with vitamin E (BS, Vitamin E, n = 6); (2) injured, nebulized with saline (BS, Saline, n = 6); and (3) not injured, not treated (Sham, n = 5). While under deep anesthesia with isoflurane, the sheep were subjected to a flame burn (40% total body surface area, 3rd degree) and inhalation injury (48 breaths of cotton smoke,40 degrees C). All groups were resuscitated with Ringer lactate solution (4 mL/kg/%burn/24 h) and placed on a ventilator [positive end-expiratory pressure (PEEP) = 5 cm H2O, tidal volume = 15 mL/kg] for 48 h. BS injury halved the lung alpha-tocopherol concentrations (0.9 +/- 0.1 nmol/g) compared with sham-injured animals (1.5 +/- 0.3), whereas vitamin E treatment elevated the lung alpha-tocopherol concentrations (7.40 +/- 2.61) in the injured animals. BS injury decreased pulmonary gas exchange (PaO2/FiO2 ratios) from 517 +/- 15 at baseline to 329 +/- 49 at 24 h and to 149 +/- 32 at 48 h compared with sham ratios of 477 +/- 14, 536 +/- 48, and 609 +/- 49, respectively. Vitamin E treatment resulted in a significant improvement of pulmonary gas exchange; ratios were 415 +/- 34 and 283 +/- 42 at 24 and 48 h, respectively. Vitamin E nebulization therapy improved the clinical responses to burn and smoke inhalation-induced acute lung injury.

Smoke inhalation increases mortality and morbidity in burn patients. We have reported that smoke inhalation increases lung lymph flow, an index of pulmonary transvascular fluid flux and decreases reflection coefficient, an index of microvascular permeability to protein. Nitric oxide has been reported to decrease microvascular permeability to protein. We hypothesize that inhaled nitric oxide decreases pulmonary microvascular hyperpermeability following smoke inhalation. Sheep were prepared for study with a chronic lung lymph fistula, Swan-Ganz, left atrial, and femoral arterial catheters. Occluders were placed on pulmonary veins to measure reflection coefficient. All animals were insufflated with 4 × 12 breaths of cotton smoke. Sheep were randomly divided into two groups: NO (injured, treated with nitric oxide (40 ppm) inhalation, n = 6) and control (injured, not treated, n = 6). Nitric oxide inhalation was started 22 h after the insult. Control animals showed an increase in lung lymph flow, and lung water content. These changes were associated with marked increase in pulmonary microvascular resistance, pulmonary artery pressure, and decrease in reflection coefficient. Nitric oxide inhalation ameliorated the above-mentioned pathological changes. The results suggest that nitric oxide inhalation has potential for beneficial effect in the treatment of patients suffering from smoke inhalation.

Carbon brushes for automotive starters are used under severe conditions of high electric current density, high contact pressure and high sliding velocity. Therefore lead has traditionally been added to brushes to improve performance and durability. Lead is an environmental hazardous substance. In the EU, the law prohibits adding lead to brushes for electric motors which is installed on new automobiles in and after January 2005. In order to develop the lead free carbon brush for starters, we analyzed the effect and selected substitutive substance of lead. Adding lead to the brush reduces the electric resistance increase of the brush in high-temperature and high-humidity atmosphere and in high-temperature atmosphere. Furthermore lead reduces the wear amount of brush. We developed the lead free brush surpassing the lead addition brush in performance and durability by addition of lead alternatives silver and zinc.

Membrane fluidity is critical for proper membrane function and is regulated in part by the proportion of unsaturated fatty acids present in membrane lipids. The proportion of these lipids in turn varies with temperature and may contribute to temperature adaptation in poikilothermic organisms. The fundamental question posed in this study was whether the unsaturation of fatty acids contributes to the ability to adapt to temperature stress inDictyostelium. First, fatty acid composition was analysed and it was observed that the relative proportions of dienoic acids changed with temperature. To investigate the role of dienoic fatty acids in temperature adaptation, null mutants were created in the two known Δ5 fatty acid desaturases (FadA and FadB) that are responsible for the production of dienoic fatty acids. ThefadBnull mutant showed no significant alteration in fatty acid composition or in phenotype. However, the disruption offadAresulted in a large drop in dienoic fatty acid content from 51·2 to 4·1 % and a possibly compensatory increase in monoenoic fatty acids (40·9–92·4 %). No difference was detected in temperature adaptation with that of wild-type cells during the growth phase. However, surprisingly, mutant cells developed more efficiently than the wild-type at elevated temperatures. These results show that the fatty acid composition ofDictyosteliumchanges with temperature and suggest that the regulation of dienoic fatty acid synthesis is involved in the development ofDictyosteliumat elevated temperatures, but not during the growth phase.

A cell-free extract of Pseudomonas sp. strain E-3 catalyzed the conversion of 9-cis-hexadecenoic acid [16:1(9c)] to 9-trans-hexadecenoic acid [16:1(9t)] in the free acid form and when 16:1(9c) was esterified to phosphatidylethanolamine (PE). The cytosolic fraction catalyzed the isomerizations of free 16:1(9c) by itself and of 16:1(9c) esterified to PE in the presence of the membrane fraction. Tracer experiments using [2,2-2H2]16:1(9c) demonstrated that the isomerization of free 16:1(9c) occurred independently of the isomerization of 16:1(9c) esterified to PE, indicating that this bacterium has two types of activities that catalyze the cis-trans isomerization of the double bond of a mono-unsaturated fatty acid.

A novel, oil-degrading bacterium (strain T1) was isolated from a hot spring in Hokkaido, Japan. It efficiently degrades different types of fats and oils, including edible oil waste. When grown in a mineral salt medium containing 1% triacylglycerol (as salad oil), hydrolysis products were 1,3- and 1,2-diacylglycerols, monoacylglycerol, and free fatty acid. However, these products were almost completely consumed during cultivation at 30 degrees C for 5 days, indicating that extracellular lipase acts randomly at different sn-positions of acylglycerols and that strain T1 has a high capacity to utilize free fatty acids. Secreted lipase activity was induced by salad oil and oleic acid. This strain was a Gram-negative straight rod shaped, aerobic, with a polar flagellum, capable of growing in temperature ranges between 15 degrees C and 55 degrees C. The 16S rRNA gene sequence analysis and DNA-DNA hybridization revealed it as a new strain of Pseudomonas aeruginosa. The type strain was T1.

Acute respiratory distress syndrome is a major complication in patients with thermal injury. The obstruction of the airway by cast material, composed in part of fibrin, contributes to deterioration of pulmonary gas exchange. We tested the effect of aerosol administration of tissue plasminogen activator, which lyses fibrin clots, on acute lung injury in sheep that had undergone combined burn/smoke inhalation injury. Anesthetized sheep were given a 40% total body surface, third degree burn and were insufflated with cotton smoke. Tissue plasminogen activator (TPA) was nebulized every 4 h at 1 or 2 mg for each nebulization, beginning 4 h after injury. Injured but untreated control sheep developed multiple symptoms of acute respiratory distress syndrome: decreased pulmonary gas exchange, increased pulmonary edema, and extensive airway obstruction. These control animals also showed increased pulmonary transvascular fluid flux and increased airway pressures. These variables were all stable in sham animals. Nebulization of saline or 1 mg of TPA only slightly improved measures of pulmonary function. Treatment of injured sheep with 2 mg of TPA attenuated all the pulmonary abnormalities noted above. The results provide evidence that clearance of airway obstructive cast material is crucial in managing acute respiratory distress syndrome resulting from combined burn and smoke inhalation injury.

Some eubacteria produce long chain polyunsaturated fatty acids (PUFAs) such as eicosapentaenoic acid (EPA) and/or docosahexaenoic acid (DHA) by the mechanism differing from the combination of elongation and oxygen-dependent desaturation of fatty acids. Metz et al. (2001) proposed a possible EPA biosynthetic pathway of bacteria, where EPA can be produced by polyketide synthase (PKS)-like enzyme encoded by the EPA gene cluster. Accordingly, a trans double bond at Δ2 of acyl intermediates is isomerized to cis configuration either at Δ3 or at Δ2, when it is not reduced to the C-C bond.

The beta-lactamase gene (mbla) of the psychrophilic marine bacterium Moritella marina strain MP-1 was identified in a previously isolated genomic DNA fragment and it was expressed in Escherichia coli cells. The mbla gene encoded a protein consisting of 287 amino acid residues. Its predicted amino acid sequence showed approximately 50% identity with that of a number of class A beta-lactamases, especially with that of CARB/PSE type of beta-lactamases (carbenicillinases). E. coli transformed with the plasmid containing mbla grew on an ampicillin-containing plate at 37 degrees C but not at 42 degrees C, suggesting that the beta-lactamase of this bacterium is heat-labile.

A possible mechanism of resistance to hydrogen peroxide (H 2 O 2 ) in Vibrio rumoiensis , isolated from the H 2 O 2 -rich drain pool of a fish processing plant, was examined. When V. rumoiensis cells were inoculated into medium containing either 5 mM or no H 2 O 2 , they grew in similar manners. A spontaneous mutant strain, S-4, derived from V. rumoiensis and lacking catalase activity did not grow at all in the presence of 5 mM H 2 O 2 . These results suggest that catalase is inevitably involved in the resistance and survival of V. rumoiensis in the presence of H 2 O 2 . Catalase activity was constitutively present in V. rumoiensis cells grown in the absence of H 2 O 2 , and its occurrence was dependent on the age of the cells, a characteristic which is observed for the HP II-type catalase of Escherichia coli . The presence of the HP II-type catalase in V. rumoiensis cells was evidenced by partial sequencing of the gene encoding the HP II-type catalase from this organism. A notable difference between V. rumoiensis and E. coli is that catalase is accumulated at very high levels (∼2% of the total soluble proteins) in V. rumoiensis , in contrast to the case for E. coli . When V. rumoiensis cells which had been exposed to 5 mM H 2 O 2 were centrifuged, most intracellular proteins, including catalase, were recovered in the medium. On the other hand, when V. rumoiensis cells were grown on plates containing various concentrations of H 2 O 2 , individual cells had a colony-forming ability inferior to those of E. coli , Bacillus subtilis , and Vibrio parahaemolyticus . Thus, it is suggested that when V. rumoiensis cells are exposed to high concentrations of H 2 O 2 , most cells will immediately be broken by H 2 O 2 . In addition, the cells which have had little or no damage will start to grow in a medium where almost all H 2 O 2 has been decomposed by the catalase released from broken cells.

Within the field of food biotechnology, microorganisms have been well utilized as bioreactors and source mechanisms for such useful biosubstances as enzymes, antibiotics, organic and amino acids, vitamins, and so on.1,2 Most microorganisms thus applied have been mesophilic. Recently, the utilization of thermophilic organisms has increased because of their thermostable characteristics. However, the utilization of cold-adapted microorganisms in food biotechnology has been very limited.3–5

We recently presented clear evidence that the major low-phosphate-inducible phosphatase of the duckweedSpirodela oligorrhiza is a glycosylphosphatidylinositol (GPI)-anchored protein, and, to our knowledge, is the first described from higher plants (N. Morita, H. Nakazato, H. Okuyama, Y. Kim, G.A. Thompson, Jr. [1996] Biochim Biophys Acta 1290: 53–62). In this report the purified 57-kD phosphatase is shown to be a purple metalloenzyme containing Fe and Mn atoms and having an absorption maximum at 556 nm. The phosphatase activity was only slightly inhibited by tartrate, as expected for a purple acid phosphatase (PAP). Furthermore, the protein cross-reacted with an anti-Arabidopsis PAP antibody on immunoblots. The N-terminal amino acid sequence of the phosphatase was very similar to those of Arabidopsis, red kidney bean (Phaseolus vulgaris), and soybean (Glycine max) PAP. Extracts of S. oligorrhiza plants incubated with the GPI-specific precursor [3H]ethanolamine were treated with antibodies raised against the purified S. oligorrhizaphosphatase. Radioactivity from the resulting immunoprecipitates was specifically associated with a 57-kD band on sodium dodecyl sulfate-polyacrylamide gels. These results, together with previous findings, strongly indicate that the GPI-anchored phosphatase ofS. oligorrhiza is a PAP.

The glycosylphosphatidylinositol (GPI)-anchored phosphatase of S. oligorrhiza plants was a purple-colored metalloenzyme containing Fe and Mn atoms and its absorption spectrum had the maximum at 556 nm. Western blot experiment using anti-A. thaliana purple acid phosphatase (PAP) antibody showed that the S. oligorrhiza phosphatase cross-reacted with the antibody. These results suggest that the GPI-anchored phosphatase is a PAP. A 1.4 kbp cDNA clone was obtained from a S. oligorrhiza cDNA library by plaque hybridization using an A. thaliana PAP gene as a probe. The estimated amino acid sequence of this clone showed 82% and 80% similarity with the sequences of the A. thaliana and Phaseolus vulgaris PAPs, respectively.

Proteins containing glycosylphosphatidylinositol(GPI) anchors (Fig. 1) are widespread in animal cells [2], where they are highly localized on the outer face of the plasma membrane. There are over 150 different GPI-anchored proteins known, with alkaline phosphatase, acetylcholinesterase, and 5’-nucleotidase being noteworthy as extensively studied examples. Recently their occurrence has been reported in archaebacteria [3] and in the green alga Chlorella saccharophila [4]. In an effort to determine whether higher plants also contain GPI-anchored proteins, we examined the aquatic duckweed Spirodela oligorrhiza because it has been reported [5] to contain alkaline phosphatase, a protein typically GPI-anchored in animals [2].Our work describing a GPI-anchored alkaline phosphatase in S.oligorrhiza [6] represents the first finding of a GPI-anchored protein in higher plants.

A novel automated C-terminal fragment peptide fractionator has been constructed. Digests with lysyl endopeptidase were covalently immobilized on p-phenylene diisothiocyanate polymer beads. Only the C-terminal fragment, which contains no lysyl residue, was liberated by cleavage at the first peptide bond of the immobilized fragment peptides with trifluoroacetic acid, and it was automatically collected. The whole procedure was automatically and precisely performed under microprocessor control in a nitrogen atmosphere. The resulting fragment was sequenced without further purification. Sequences of both N- and C-terminal regions can be routinely obtained for ca. 100 pmol samples by the use of a conventional automated Edman-type protein sequencer.

There are now more than 100 different proteins known to be attached to the cell surface of animal or yeast cells by a glycosylphosphatidylinositol(GPI) anchor [1]. The structure of these anchoring units (Fig 1) is relatively highly conserved, although additional sugar moieties are sometimes attached to the basic oligoglycan backbone, and the diacylglycerol lipid unit is in certain molecules replaced by an alkyl,acyl-glycerol or a ceramide structure. Among the more familiar proteins now known to be GPI-anchored are alkaline phosphatase, acetylcholinesterase, and the variable surface glycoprotein coating the surface of trypanosomes. Surprisingly, there have been no reports of GPI-anchored proteins in plants. Because my laboratory has an ongoing project to study the GPI-anchored proteins of the protozoan Tetrahymena, we have been able to apply this technology in a search for these proteins in higher plants.

Saposin-C, a small acidic glycoprotein that can activate glucosylceramide-s-glucosidase, has been isolated from bovine spleen. The complete amino acid sequence of bovine saposin-C was determined by Edman degradation of the purified protein and its fragmented peptides. It contains 80 amino acids, one carbohydrate chain attached to a single aspargine residue and six cysteine residues in oxidized form. The sequence of bovine saposin-C is 76 and 65% identical with the sequences of saposin-C from human spleen and guinea pig liver, respectively. Hydropathy profiles of the sequence of saposin-C from three species were similar despite the significant residue substitutions. Bovine saposin-C had a stronger effect in stimulating bovine s-glucosidase compared to human saposin-C. However, the effect of human saposin-C in stimulating human enzyme was stronger than that of bovine saposin-C. The region around residue 35, which is next to the extremely hydrophilic region, seems to be important to produce an interaction with the enzyme.

Vibrio sp. strain ABE-1 is a unique marine bacterium in terms of its ability to synthesize Δ9-trans-hexadecenoic acid and Δ7-cis-tetradecenoic acid (14:1(7c); Okuyama, H., Sasaki, S., Higashi, S. and Murata, N. (1990) J. Bacteriol. 172, 3515-3518). The present study, involving labeling with [1-14C]acetate, demonstrated that 14:1 is synthesized by the anaerobic pathway. When cells of this bacterium were grown in the presence of [1-14C]myristic acid (14:0), this compound was converted to palmitic (16:0) and hexadecenoic (16:1) acids but not to 14:1, under aerobic conditions. These results suggest that the incorporated 14:0 was elongated to 16:0 and then converted to 16:1 by the aerobic desaturation of 16:0. It appears that the anaerobic pathway and aerobic desaturation are both involved in the synthesis of unsaturated fatty acids during aerobic growth of Vibrio sp. strain ABE-1.

One of the ways to improve the heating efficiency of a heater is to eliminate the passivation layer of the heater by using TaSiO for the heater material, whereby the heater surface becomes thermally oxidized. Rapid boiling of water occurs at around 300°C; therefore, it is necessary for the fabrication condition of the oxidation layer to correspond to a temperature higher than the boiling temperature. In a pulse drive during printing, it is also necessary to keep the heat at a temperature lower than that of thermal oxidization to prevent deterioration. As a result of observing the generation of bubbles and measuring the heater temperature, it was confirmed that within the range of heating velocity in this experiment, boiling starts at around 300°C, and it was also possible to understand pulse control in thermal ink jet printheads to maintain the heater at an appropriate temperature.

When docosahexaenoic acid (DHA)-producing Moritella marina strain MP-1 was cultured in the medium containing 0.5 microg cerulenin ml-1, an inhibitor for fatty acid biosynthesis, the cells grew normally, but the content of DHA in the total fatty acids increased from 5.9-19.4%. The DHA yield of M. marina strain MP-1 cells also increased from 4 to 13.7 mg l-1 by cerulenin treatment. The same effect of cerulenin was observed in eicosapentaenoic acid (EPA)-producing Shewanella marinintestina strain IK-1 grown in the medium containing 7.5 microg cerulenin ml-1, and the cerulenin treatment increased the EPA yield from 1.6 to 8 mg l-1. The use of cerulenin is, therefore, advantageous to increase the content of intracellular polyunsaturated fatty acids (PUFA) in particular PUFA-containing phospholipids in bacterial cells.

We reported that the bronchial circulation contributes to pulmonary function after acute lung injury with smoke inhalation. But the observation period was 24 hours. We hypothesize that the bronchial circulation plays a major role in the lung injury seen with the combination of burn and smoke inhalation injury.

Introduction. α-Tocopherol (Vit. E) plays a pivotal role in clinical settings associated with oxidative stress. In patients suffering from burn and smoke inhalation injury, a decrease in Vit. E plasma level is typically linked with increased mortality rates. Previously we have shown that oral administration of α-tocopherol ameliorates the acute lung injury in sheep with burn and smoke inhalation injury. However, thermal injury often associates with ileus. In the present study we tested the effect of aerosolized Vit. E nebulized into the airway. Method. Sixteen adult ewes, weighing ∼33 kg, were operatively instrumented for chronic study. The sheep were randomly allocated to groups (a) noninjured, nontreated, (b) injured, nontreated, (c) injured, nebulized with Vit. E, and (d) injured, orally administered with Vit. E. The sheep were subjected to a third-degree flame burn (40% of total body surface) and smoke inhalation (cotton smoke). All groups were placed on the ventilator and resuscitated similarly. Results. Pulmonary dysfunction in injured nontreated group was associated with significant depletion of both α- and γ-tocopherol. Compared with injured controls, the Vit. E nebulization group had lung tissue Vit. E levels which were significantly increased (∗ P 2 /FiO 2 ratio at 48 h postinjury (Control, 93 ± 15.9; Oral Vit. E, 232 ± 31.8; Nebulized, 305 ± 30.9), and pulmonary microvascular permeability index (Control, 27.53 ± 10.29; Oral Vit. E 14.30 ± 4.85; Nebulized, 8.68 ± 2.218) in these animals. Conclusions. The role of Vit. E on pulmonary antioxidant defense system has been known. Previously there was no effective technique to administer Vit. E directly into the injured organ. We think that aerosolized Vit. E nebulization will be a good treatment strategy for patients with smoke inhalation and burn.

The MWD (measuring while drilling) system, which transmits information such as temperature, pressure, direction, and inclination at the bottom of the hole to the surface of the earth while drilling, is used and is underdeveloped as a higher temperature version, but it was hoped that it would be more efficient in the oil fields and the geothermal fields as a supporting tool in drilling. The transmission from MWD is realized by the pulse interval modulation, which generates pulsed pressure waves and uses the interval of pressure waves to send physical information. This paper shows that our calculation results can accurately simulate the mud pulse in drill strings comparing them with experiments carried out in the laboratory and that, due to the loss of propagation depending upon well depths, the behavior of mud pulse signals apparently change. It is concluded that the simulation program is reliable for this system, and that the mud signal is useful from a 500‐ to 3000‐m depth borehole.