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Production of chiral R-3-hydroxyalkanoic acids and R-3-hydroxyalkanoic acid methylesters via hydrolytic degradation of polyhydroxyalkanoate synthesized by pseudomonads.

A novel and efficient method for the production of enantiomericaly pure R-3-hydroxyalkanoic acids and R-3-hydroxyalkanoic acid methylesters was developed. The described method is based on hydrolysis of poly(hydroxyalkanoate) copolymers synthesized by Pseudomonas putida. The polymer was isolated via solvent recovery and hydrolyzed by acid methanolysis. The obtained 3-hydroxyalkanoic acid methylester mixture was distilled into several fractions with an overall yield of 96.6% (w/w). Gas chromatography-mass spectrometry analysis of the fractions showed that 3-hydroxyhexanoic-, 3-hydroxyoctanoic-, 3 hydroxydecanoic-, and 3-hydroxydodecanoic acid methylesters were enriched to purities exceeding 96 mol%, with distillation yields of 99.9, 99.8, 88.4, and 56.8% (w/w), respectively. Subsequent saponification of the purified methylester fractions yielded the corresponding 3-hydroxyalkanoic acids, which were recovered up to 92.8% (w/w). Chiral gas chromatography analysis confirmed that both 3-hydroxyoctanoic acid and 3-hydroxyoctanoic acid methylester are present in the R-form at a very high enantiomeric excess (>99.9%).

Fermentation↗

Screening antimicrobial activity of various extracts of Artemisia dracunculus L.

The antimicrobial activities of chloroform, acetone and two different concentrations of methanol extracts of Artemisia dracunculus L. were studied. These extracts were tested against nine bacteria and four yeasts strains by the disc diffusion method. The results indicated that the methanol extract of A. dracunculus is more effective against tested microorganisms than chloroform or acetone extracts. The chloroform and acetone extracts were inhibitory only towards Pseudomonas aeruginosa (ATCC 27853). While the methanol extract that was diluted with 10 ml distilled water showed inhibition zones against Shigella (RSHI), Listeria monocytogenes ATCC 7644, P. aeruginosa (ATCC 27853), the methanol extract that was diluted with 5 ml distilled water showed inhibition zones against two different strains of Escherichia coli (RSHI, ATCC 25922), Shigella (RSHI), L. monocytogenes (ATCC 7644), and P. aeruginosa ATCC 27853. The cells of microorganisms treated with plant extracts and normal microorganism cells were observed by scanning electron microscope. It was apparent that cells are damaged after treatment with A. dracunculus.

Acetone↗

Analysis of high-frequency rotational angioplasty-induced echo contrast.

During high-frequency rotational angioplasty (HFRA), myocardial contrast enhancement (echo contrast) was observed by means of two-dimensional echocardiography. In order to evaluate the echo contrast produced by HFRA, an in vitro experimental model was built using a cylinder with the HFRA catheter scanned in a water bath. The cylinder was filled with blood, mineral water, and distilled water. With a 2.5-MHz transducer, echograms were recorded and analyzed by video intensitometer, and the maximum intensity IUmax and persistence time P were calculated. Rotation frequencies of 20,000-200,000 rpm with 2.0--mm burrs and times of 10 s were tested. In another model, hyperbaric conditions for the same media were produced. The influence of debris from arterioscleroic plaque and of hematocrit on echo contrast intensity were also analyzed. The effect of HFRA on hemolysis (LDH, free hemoglobin) within 10 s and temperature were also measured. The contrast effect was transient, depending on the rotation frequency. In blood, it appeared at 20,000 rpm (IUmax at 200,000 rpm: 237 IU), in mineral water at 40,000 rpm (IUmax 165 IU), and in distilled water at 80,000 rpm (IUmax 72 IU). Persistence time was measured up to about a half-minute. Echo contrast production was reduced at 0.5 bar and fully suppressed at 2.5 bar. Debris increased contrast intensity from about 219 IU to 225 IU (at 160,000 rpm). In blood IUmax decreased from 227 IU to 97 IU by lowering the hematocrit from 44.2% to 3.6%.(ABSTRACT TRUNCATED AT 250 WORDS)

Echocardiography↗

Effects of number of pre-exposures on sucrose taste aversion in weanling rats.

Weanling rats, 20 days old, received 0, 1, 2, 4, or 8 pre-exposures to 12% sucrose prior to a single pairing of sucrose with an intraperitoneal injection of lithium chloride (LiCl; 3.0 mEq of a .15Msolution) or distilled water (20 mg/kg). Testing for sucrose taste aversion with a 2-bottle-choice procedure on 9 test trials reliably showed that increasing numbers of pre-exposures to sucrose directly attenuated taste aversion effects in the LiCl-injected groups but did not appreciably affect intake performance in the distilled water-injected groups. Comparisons between the injection conditions yielded reliable evidence for sucrose taste aversion at each pre-exposure level. These results show that flavor stimulus pre-exposures reliably attenuate subsequent taste aversion in weanling rats as had previously been reported for adult rats.

Animals↗

Membrane active plasma factor in multiple sclerosis: characterization and isolation by recycling isoelectric focusing.

Recycling isoelectric focusing is a rapid, high resolution technique that has the capability of fractionating complex mixtures of proteins on a preparative scale on the basis of their isoelectric points (pIs). For this reason, it appeared to be an ideal tool to further characterize and isolate the surface active plasma component(s) which is abnormal in multiple sclerosis (MS). The normal control and the abnormal MS plasma components, or factors, proved to be stable under the conditions used in this technique, including deionization by electrodialysis, dialysis against distilled water, lyophilization and the presence of 3M urea and carrier distilled water, lyophilization and the presence of 3M urea and carrier ampholytes. The presence or absence of plasma factor activity was determined by incubating red blood cells in a test plasma, or plasma fraction, followed by the determination of the red blood cell electrophoretic mobility in the presence and absence of linoleic acid. Both the normal and MS factor had a pI of 4.0 +/- 0.1 under the conditions used. A high degree of purification was achieved and albumin was eliminated as a possible candidate for the factor(s).

Dialysis↗

Sterol composition of Bulgarian soya and corn oils.

The free sterols, the sterol esters and the sterol glycosides of the raw soya and corn oils as well as those of the technical lecithin and the deodorizer distillated of the latter oils were isolated by preparative TLC. The composition of each of the isolated sterol derivatives was determined by GLC and MS. Sitosterol, campesterol, stigmasterol and an unknown sterol with a molecular weight of 428 are contained in almost all of the examined fractions of the soya oil and its refinement byproducts. Dehydrocampesterol is present in the free sterols of the raw soya oil and the soya lecithin. Stigmasterol is contained in the soya deodorizer distillate in high amounts. It was established that cholesterol was present in the sterol esters of the raw soya oil high amounts. Delta7-stigmastenol occurs only in the sterol esters of the latter oil. Sitosterol, campesterol and stimgasterol are the main components of all sterol fractions of the corn oil and its refinement products. Dehydrocampesterol and unknown sterols with molecular weights of 428 are present in the free sterols of the raw corn oil. Some sterol glycosides of the soya and corn lecithin are esterified with the same major fatty acid components of the glycerides, palmitic acid being the main one. The fatty acid compositon of sterol esters of the raw soya and corn oil roughly corresponds to the fatty acid composition of oils.

Chromatography, Gas↗

[Analysis of volatile nitrosamines in food of animal origin].

A method is described for the fluorescence-spectrophotometric densitometric determination of nitrosamines. For this purpose, foods of animal origin are vacuum-distilled, the distilled-over nitrosamines are extracted with methylene chloride from an acid medium and, after cleavage by hydrogen bromide, the corresponding amines are reacted with 7-chloro-4-nitrobenzo-2-oxa-1,3-diazole. The purificaton of the extract and the resolution of the corresponding fluorescent amine derivatives are effected simultaneously by two-dimensional thin-layer chromatography. The determination is achieved by in situ fluorescence spectrophotometry against test substances on the same slab. This method seems useful for investigations into the formation of nitrosamines in foods of animal origin because it permits to detect nitrosamine concentrations as low as 0.1 microgram/kg of food.

Animals↗

[Effect of aromatic substances on the quality of wines and spirits].

The influence of technological and other factors such as yeast strain, guidance of fermentation and storage on the quality of Bulgarian wines and wine distillates was investigated with the help of sensory, chemical-analytical and chromatographical methods. The total content of aldehydes, higher alcohols or esters or the total content of volatile components estimated by GC seem to be less suitable as quality index, whereas the ratio of esters to aldehydes and the ratio of isobutanol to isoamyl alcohol are parallel to a good sensory quality. The ester content of wines increases during storage, especially in casks. With wine distillates there is a good correlation between the time of storage and the content of aromatic aldehydes.

Acids↗

Modified determination of 2-thiobarbituric acid value in fats and oils.

Several methods for the determination of TBA-value were compared with a new direct modification (omitting the previous distillation), using 1-butanol as the sole solvent. The absorbance is measured at 530 nm after heating to 95 degrees C for 120 min, and cooling. The effect of variables was tested by the Youdens ruggedness test (reaction time, temperature, cooling, concentration of sample, thiobarbituric acid, water, heavy metals, antioxidants, chelating agents, bubbling with oxygen or nitrogen). The Lambert-Beers law is valid in the absorbance range between 0.05 and 0.8. The repeatability is better than that of the Dzikowskis procedure and comparable with those of more complicated distillation methods. The standard deviation is in a linear relation with the absorbance (range 0.1-1.0). The method was found satisfactory for evaluating lard, cooking fat, soybean, sunflower, and rapeseed oils in the stage of beginning rancidity.

Antioxidants↗

Heat resistance of Bacillus spores exposed to food processing conditions.

The heat resistance of the spores of 5 Bacillus species was studied in distilled water and pasteurized skim milk. The spores were also subjected to frying and cooking conditions used in fried rice preparations. In presence of milk, the heat resistance of all the five Bacillus species was found to decrease compared to distilled water. Spores of all five Bacillus survived cooking conditions of rice. Frying and subsequent cooking conditions inactivated the spores of all five Bacillus species.

Animals↗

Skin-tumor initiation activity of coal liquids with different boiling-point ranges.

High-boiling coal liquids from the solvent-refined coal-I and -II (SRC-I, -II) processes, respectively, were fractionally distilled. In the case of SRC-I process solvent (PS), 50 degrees F distillation cuts were obtained between 550 and 850 degrees F, while for the SRC-II material, the 50 degrees F cuts were only obtained between 700 and 850 degrees F. These cuts, as well as the parent material, were tested for their ability to initiate skin tumors by applying a single dose (25 mg) to the shaved backs of Charles River female CD-1 mice. After 2 weeks, the mice received twice weekly applications of 5 micrograms of the promoter, phorbol myristate acetate. Only a few tumors were found for SRC-I fractions boiling below 700 degrees F; tumor-initiating activity increased as the boiling point increased. A similar increase in response with increasing boiling point was seen for the SRC-II cuts. The initiating activities for the parent materials were similar to those observed for their respective 800 to 850 degrees F cuts.

Animals↗

Decrease in hepatotoxicity by lead exposure is not explained by its mitogenic response.

The present research was conducted to evaluate the effect of mitogen pre-exposure on CCl4-induced hepatotoxicity. Male Wistar rats were administered a single i.p. injection of CCl4 (0.3 ml kg-1 in corn oil) 48 h following either a single dose of lead nitrate (0.33 mg kg-1) or distilled water via i.v. injection. Hepatotoxicity, as measured by serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels, was monitored 6, 24, 48, 72 and 120 h after CCl4 exposure. The lead nitrate-pretreated rats displayed markedly lower serum ALT and AST levels at 24, 48 and 72 h than rats pretreated with distilled water. However, treatment with the antimitotic agent colchicine did not alter the lead-induced protection. These findings suggest that the lead-induced protection is not associated with the major mitogenic response of lead, despite its strong temporal association. A critical review of the available toxicological data also argues against the lead protection being a function of its capacity to inhibit cytochrome P-450.

Alanine Transaminase↗

In vitro stability of a novel compliant poly(carbonate-urea)urethane to oxidative and hydrolytic stress.

Poly(ester)urethane and poly(ether)urethane vascular grafts fail in vivo because of hydrolytic and oxidative degradative mechanisms. Studies have shown that poly(carbonate)urethanes have enhanced resistance. There is still a need for a viable, nonrigid, small-diameter, synthetic vascular graft. In this study, we sought to confirm this by exposing a novel formulation of compliant poly(carbonate-urea)urethane (CPU) manufactured by an innovative process, resulting in a stress-free. Small-diameter prosthesis, and a conventional poly(ether)urethane Pulse-Tec graft known to readily undergo oxidation in a variety of degradative solutions, and we assessed them for the development of oxidative and hydrolytic degradation, changes in elastic properties, and chemical stability. To simulate the in vivo environment, we used buffered solutions of phospholipase A(2) and cholesterol esterase; solutions of H(2)O(2)/CoCl(2), t-butyl peroxide/CoCl(2) (t-but/CoCl(2)), and glutathione/t-butyl peroxide/CoCl(2) (Glut/t-but/CoCl(2)); and plasma fractions I-IV, which were derived from fresh human plasma centrifuged in poly(ethylene glycol). To act as a negative control, both graft types were incubated in distilled water. Samples of both graft types (100 mm with a 5.0-mm inner diameter) were incubated in these solutions at 37 degrees C for 70 days before environmental scanning electron microscopy, radial tensile strength and quality control, gel permeation chromatography, and in vitro compliance assessments were performed. Oxidative degradation was ascertained from significant changes in molecular weight with respect to a control on all Pulse-Tec grafts treated with t-but/CoCl(2), Glut/t-but/CoCl(2), and plasma fractions I-III. Pulse-Tec grafts exposed to the H(2)O(2)/CoCl(2) mixture had significantly greater compliance than controls incubated in distilled water (p < 0.001 at 50 mmHg). No changes in molecular weight with respect to the control were observed for the CPU samples; only those immersed in t-but/CoCl(2) and Glut/t-but/CoCl(2) showed an 11% increase in molecular weight to 108,000. Only CPU grafts treated with the Glut/t-but/CoCl(2) mixture exhibited significantly greater compliance (p < 0.05 at 50 mmHg). Overall, results from this study indicate that CPU presents a far greater chemical stability than poly(ether)-urethane grafts do.

Biocompatible Materials↗

Dynamic mechanical properties of medical grade silicone elastomer stored in simulated body fluids.

Medical grade silicone rubber samples were immersed in distilled water and in pseudo-extracellular fluid (PECF) at 37.5 degrees C for up to four months. Periodically, the storage modulus and log decrement of the samples were measured on a torsion pendulum. Up to 20% increases in storage modulus and about 40% decreases in log decrement were found over the four months independent of sample surface area. Similar trends were seen in both distilled water and in PECF. Higher (59 degrees C) storage temperatures accelerated the changes. Small amounts of moisture uptake were detected. However, the presence of water in the samples did not cause the change in mechanical properties.

Biomechanical Phenomena↗

Surface modification and characterization of some commonly used catheter materials. II. Friction characterization.

The effects of the modification of polystyrene (PS), polyethylene (PE), poly(vinyl chloride) (PVC), silicone rubber (SR), and fluorinated ethylene propylene (FEP) copolymer by radio frequency glow discharge in a helium environment were presented in part I. The hydrated polymer surfaces were characterized by XPS, SEM, visual microscopy, and by contact angle measurements. In general, exposure of the polymers to RFGD produced an oxidized hydrophilic surface, yet the roughness of the surface was unaltered by the relatively mild plasma conditions used. In this article, the frictional behavior of oxidized and unoxidized SR, PE, and FEP in distilled water, isotonic saline, and blood plasma environments is examined experimentally. The results are discussed in relation to the properties generally believed to affect frictional phenomena and to the surface properties as determined in part I. Results indicate that RFGD-treated SR generates less friction than untreated SR when dragged across all untreated and treated polymer surfaces, whether the medium is distilled water or an isotonic saline solution. Friction is consistently lower in a blood plasma medium between all surfaces investigated, most probably because of the presence of adsorbed proteins at the polymer interfaces.

Biocompatible Materials↗

Influence of storage regime prior to abrasion on surface topography of restorative materials.

This investigation was carried out to evaluate the effect of storage conditions prior to brushing simulation on surface texture of restorative materials. One resin-modified glass ionomer (Fuji II LC Improved/GC Corp.), one polyacid-modified composite resin (Dyract AP/Denstply), one microfill composite (Durafill VS/Kulzer), and one hybrid (Filtek-Z250/3M) composite were tested. Forty-five standardized cylindrical specimens of each material were made and randomly divided into three groups according to their subsequent storage conditions: distilled deionized water, artificial saliva, or pH-cycling regime. After 24 h, the experimental units were finished and polished and the surface roughness was measured to obtain Ra baseline values (Bv). Samples were subjected to their assigned storage regime and brushed afterwards. By the end of 10 repetitions of this protocol, final surface roughness readings (Fv) were taken. The analysis of covariance (alpha = 0.05), considering the covariate Bv showed a significant interaction between restorative material and storage condition (p(value) = 0.0002). Tukey's test revealed that the pH-cycling model provided a significantly lower surface roughness for Fuji II LC and Dyract AP than did the other media. For both composites no significant difference among storage regimes was detected. Under a condition simulating dynamic variation in pH prior to abrasion, the resultant surface texture may be either smoothed down or unchanged, depending on the restorative material, when compared to the effect provided by artificial saliva and distilled deionized water.

Biocompatible Materials↗

Leaching and mechanical properties characterization of dental composites.

The objective of this study was to determine the leaching of Ba, Si, and Sr from four dental composites: Restolux (RX), Micronew (M), Renew (RW), and Choice (C) and to correlate the effects of such leaching with flexure strength and modulus of elasticity. The specimens were 3 x 3 x 25-mm bars, polished with 120- and 240-grit SiC paper, and were aged for 4, 6, and 8 months in artificial saliva, distilled water, and a 50/50 mixture of ethanol and distilled water. Inductively coupled plasma with mass spectroscopy (ICP/MS) was used to determine the ion concentrations after aging. The greatest release of Ba and Sr occurred after aging in 50/50 volume fraction mixture of ethanol/water and for Si in artificial saliva. The 50/50 solution caused a breakdown of the resin and glass matrix, resulting in an increase of Ba and Sr, whereas aging in artificial saliva resulted in an ion charge balance, which allowed an elevated and continuous release of Si. The flexure strength and the elastic modulus showed a gradual decrease over time, with the greatest decreases occurring following aging in the 50/50 water/ethanol solution, but little correlation with the leaching of the filler ions.

Biocompatible Materials↗

Environmental degradation of glass-ionomer cements: a depth-sensing microindentation study.

This study investigated the effects of various environmental conditions on the hardness and elastic modulus of restorative glass-ionomer cements (GICs). Two resin-modified GICs (RMGICs) (Fuji II LC [FL]; Photac-Fil Quick [PQ]) and three highly viscous GICs (HVGICs) (Fuji IX Fast [FN]; KetacMolar [KM]; KetacMolar Quick [KQ]) were evaluated in this study. Specimens were fabricated according to the manufacturers' instructions and stored under a variety of conditions (n = 7): 100% humidity, distilled water, pH 5 demineralization solution, and pH 7 remineralization solution. The hardness and elastic modulus were measured using a depth-sensing microindentation test after 4 weeks. The results were analyzed using the independent samples T-test and ANOVA/Scheffe's post hoc test (p < 0.05). HVGICs showed significantly higher hardness and elastic modulus than RMGICs under all storage conditions. Storage in distilled water significantly increased the hardness and elastic modulus of FN, but decreased that of PQ. All HVGICs and RMGICs stored in remineralization solution had hardness values and elastic moduli comparable to those stored in water. Compared to remineralization solution, demineralization solution had no significant effects on the modified GICs with the exception of KQ. The results suggest that the mechanical properties of glass-ionomer restoratives are material-type and storage condition dependent. Therefore, the clinical selection of a glass-ionomer material should be based on the oral environment to which it will be subjected.

Biodegradation, Environmental↗