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Previous workers from this laboratory observed considerable variation in the proportions of acetic and lactic acids produced in pure broth culture as compared to consistently high proportions of acetic acid produced in the sourdough and flour suspension systems. In the latter the proportion of acetic acid was always in the range of 20 to 35% of the total, whereas in pure broth culture frequently less than 5% acetic acid was produced. In the natural environment, the sourdough bacteria, tentatively identified as lactobacilli, coexist with a yeast, Saccharomyces exiguus, and this study was undertaken to determine whether this yeast or flour ingredients including glucose or other factors were involved in this variable production of acetic acid. The proportion of acetic acid produced in broth culture on maltose, the preferred carbohydrate source, was found to depend almost entirely on the degree of aeration. Essentially anaerobic conditions, as obtained by thorough evacuation and flushing with CO(2) or N(2), resulted in very low (5% or less) proportions of acetic acid. Aerobic conditions, achieved by continuous shaking in cotton-plugged flasks, yielded high levels (23 to 39% of the total) of acetic acid. Similar effects of aeration were observed with glucose as the substrate, although growth was considerably slower, or in nonsterile flour suspension systems. It is theorized that, under aerobic conditions, the reduced pyridine nucleotides generated in the dissimilation of carbohydrate are oxidized directly by molecular oxygen, thereby becoming unavailable for the reduction of the acetyl phosphate intermediate to ethyl alcohol, the usual product of anaerobic dissimilation of glucose by heterofermentative lactic acid bacteria. Comparative studies with known strains of homo- and heterofermentative lactobacilli showed similar effects of aeration only on the heterofermentative strains, lending additional support to the tentative grouping by previous workers from this laboratory of the sourdough bacteria with the heterofermentative lactobacilli.
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PMID:Factors affecting organic acid production by sourdough (San Francisco) bacteria. 504 65

To develop an efficient method for the production of l-citrulline, optimum conditions for the conversion of l-arginine to l-citrulline by microbial l-arginine deiminase and for production of the enzyme were studied. A number of micro-organisms were screened to test their ability to form and accumulate l-citrulline from l-arginine. Pseudomonas putida was selected as the best organism. With this organism, enzyme activity as high as 9.20 units per ml could be produced by a shaking culture at 30 C in a medium containing glucose, ammonium phosphate, l-arginine hydrochloride, yeast extract, peptone, and inorganic salts. Appropriate addition of a surface active agent to the reaction mixture was found to shorten the time required for the conversion. A large amount of l-arginine hydrochloride was converted stoichiometrically to l-citrulline in 62 hr at 37 C. Accumulated l-citrulline was readily isolated in pure form by ordinary procedures with ion-exchange resins. Yields of isolated l-citrulline of over 90.5% from l-arginine hydrochloride were easily attainable.
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PMID:Enzymatic production of L-citrulline by Pseudomonas putida. 513 89

Pasteurella pestis, harvested after 24 to 30 hr of growth in a casein hydrolysate medium at 26 C, was resuspended and shaken in 3% lactose-0.1 m phosphate buffer for 4 hr at the same temperature. Certain characteristics of these starved cells were compared with those of control cells. No differences in the amounts of cellular carbohydrate or lipid were detected. The concentrations of the principal free amino acids were greater in the shaken cells, except that they contained no measureable arginine, and the normally large pools of intracellular tricarboxylic acid cycle intermediates were reduced. Greater viable-cell counts resulted with the cells that were shaken in lactose buffer than with the control cells when each was incubated at 5 C for several weeks. However, the reduced viabilities were apparent losses caused by the formation of aggregates of cells. The clumping of cells was caused by the polymerization of extracellular nucleic acids, principally deoxyribonucleic acid, that were excreted by the cells. Cell clumping could be partially prevented by prior shaking of the suspended cells, which removed some of the deleterious material, or by the action of crystalline deoxyribonuclease.
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PMID:Biochemical and physical changes in shaken suspensions of Pasteurella pestis. 533 54

Cells of Arthrobacter crystallopoietes, harvested during growth as spheres and as rods, were starved by shaking at 30 C in phosphate buffer for 30 days, during which time they maintained 100% viability. Changes in cellular components and the activity of specific enzyme pathways were monitored. A glycogen-like polysaccharide comprised 40% of the dry weight of growing spherical cells and 10% of the dry weight of rod cells. This material was utilized at approximately the same rate, on a percentage basis, during starvation of both cell forms. The rods degraded intracellular protein at approximately twice the rate of the spheres. At the end of 30 days, the rods had degraded 40% and the spheres 20% of their initial content of protein. Ribonucleic acid (RNA) was degraded significantly more rapidly in the rods. After 30 days starvation, 85 and 32% of the initial RNA of rods and spheres, respectively, had been depleted. Magnesium ion followed this same general pattern; the rods lost 65% and the spheres 45% of their initial content during 28 days of starvation. Deoxyribonucleic acid increased by 20% during the first few hours of starvation of both cell forms and then remained constant. The ability of glucose-, succinate-, and 2-hydroxypyridine (2-HP)-grown cells to oxidize glucose remained constant during 14 days of starvation. The ability of succinate-grown cells to oxidize succinate decreased rapidly during the first few hours of starvation to a rate which remained constant for 14 days. Cells adapted to growth on 2-HP completely lost their ability to oxidize this substrate after 3 days starvation.
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PMID:Intracellular substrates for endogenous metabolism during long-term starvation of rod and spherical cells of Arthrobacter crystallopoietes. 547 76

The formation of adenosine cyclic 3',5'-phosphate by Brevibacterium liquefaciens ATCC 14929 was studied with the use of nonproliferating cells and cell-free extract. With nonproliferating cells provided by deprivation of sulfate, the formation of this nucleotide was accelerated by adding some amino acids and sugars. Among amino acids tested, alanine and asparagine were most effective. Pentoses were more favorable than hexoses and other sugars. Formation of adenosine cyclic 3',5'-phosphate was observed also with chloramphenicol-treated cells. Experiments on cell-free extract showed that addition of alanine or pyruvate stimulated the formation of adenosine cyclic 3',5'-phosphate from adenosine-5'-triphosphate. When alanine was added to the cell-free system, shaking of the reaction mixture further increased the amount of the nucleotide, but pyruvate was far more effective than alanine. No synergistic effect of alanine and pyruvate was observed. Some enzyme activity was observed which decomposed adenosine cyclic 3',5'-phosphate, but it was weak as compared with adenyl cyclase activity in the presence of pyruvate. From the results obtained, it appears that pyruvate may act as an activating factor of adenyl cyclase in Brevibacterium liquefaciens.
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PMID:Formation of adenosine cyclic 3',5'-phosphate by nonproliferating cells and cell-free extract of Brevibacterium liquefaciens. 603 54

A method of fixed rabies virus purification from infected sheep brains was proposed. It consisted of suspending the brain tissue in phosphate-buffered hypertonic (0.3 M) NaCl solution, shaking at 37 degrees C and low speed centrifugation at the same temperature. From 65 to 85% ballast proteins were removed, the neuroallergenic activity of the material was lost, but practically no losses of the virus occurred.
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PMID:Freeing rabies virus of the neuroallergenic factor from brain tissue. 611 64

Phosphorylated proteins may play an important role in regulating the metabolism or function of rod photoreceptors. In mammalian retinas, a photoreceptor protein of 33 000 (33K) molecular weight is phosphorylated in a cyclic nucleotide dependent manner in vitro. Since light initiates the activation of a photoreceptor-specific phosphodiesterase and a rapid reduction in guanosine cyclic 3',5'-phosphate concentration, phosphorylation of the 33K protein may be modulated by light in situ. In order to test this possibility, dark-adapted rat retinas were incubated for 30 min in the dark in phosphate-free Kreb's buffer containing [32P]orthophosphate. Following incubation, rod outer segments were detached by shaking, and the 32P-labeled rod outer segment proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, detected by autoradiography, and quantitated by densitometric scanning. The incorporation of radioactivity (32P) into the 33K protein was higher than into any other rod outer segment protein, and the amount of 32P-labeled 33K protein in the detached rod outer segments remained unchanged during 10 additional min of darkness. The addition of isobutylmethylxanthine to the incubation medium enhanced the incorporation of 32P into 33K protein to about 400% of the original level. Exposure of freshly detached rod outer segments to room light for 90 s decreased the amount of labeled 33K protein to 45% of its original level. The dephosphorylation of labeled 33K protein continued, reaching 12% of the original dark value 10 min after the previously illuminated sample was returned to darkness. Light initiated the phosphorylation of rhodopsin, and rhodopsin phosphorylation continued during the postillumination period of darkness.
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PMID:Light-induced dephosphorylation of a 33K protein in rod outer segments of rat retina. 620 20

A micro-enzyme-linked immunosorbent assay (ELISA) utilizing antigen from lyophilized Clonorchis sinensis adult worms extracted with triethanolamine buffered saline (TBS), veronal buffered saline (VBS) or phosphate buffered saline (PBS) were used to test 66, 31 and 9 sera from clonorchiasis patients, uninfected normal humen and patients with other parasitic infections, respectively. Four different combinations of incubation time between serum with antigen and serum with conjugate at either 37 degrees C or room temperature were tried. The combination of 6 hours and 16 hours reaction time processed at room temperature with shaking was chosen in this study because it revealed no pseudo-negative and cross reactions with high sensitivity, specificity and predictive value. There were no statistic difference among mean optical densities in micro-ELISA reactions of 66 sera (in 800 X dilution) from clonorchiasis patients with TBS, VBS or PBS antigen, respectively, but results obtained from TBS antigen seemed better than other two antigens. Good correlation was observed between the antibody titer or ELISA readings of sera form clonorchiasis patients and their worm burden expressed as eggs per gram of feces (EPG).
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PMID:[Immunodiagnosis of human clonorchiasis by a micro-enzyme-linked immunosorbent assay]. 653 54

The optimal conditions for preparations of rifampicin-containing liposomes were determined with the methods of mechanical shaking, gas dispersion and and reversible phases. It was found that the percentage of rifampicin incorporation into liposomes depended on the molar ratio of the antibiotic to the lipid (the optimal ratio was 1 : 10), the size and structure of liposomes, the amount of cholesterol added and the lipid membrane charge. Incorporation of rifampicin amounted to 16.1 +/- 2.4, 39.2 +/- 3.2 and 60.5 +/- 2.9 per cent with respect to neutral lecithin multilamellar liposes, liposomes prepared with the gas dispersion method and liposomes prepared with the method of reversible phases, respectively. Cholesterol in a molar ratio to lecithin equal to 2 : 5 or higher and dicetyl phosphate imparting the negative charge to the membrane had an inhibitory effect on the drug uptake by liposomes, while stearyl amine having the positive charge had a stimulating effect. The effect of the cryoprotectors glucose, polyvinylpyrrolidone, poly-ethylene glycole-400 and glycerol on low-temperature preservation and storage of rifampicin-containing liposomes was studied. It was shown that 10--15 per cent solutions of sucrose and glucose had the highest cryoprotective effect, when the two-stage method of freezing was used. It provided almost 84 per cent preservation of liposomal rifampicin. Electron microscopy showed that after defrosting liposomes no significant changes in the size and structure of lipid membranes were observed.
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PMID:[Isolation and low-temperature preservation of liposomes containing rifampicin]. 663 70

Albuterol is a long-acting beta 2-adrenergic receptor-selective drug that relaxes airway smooth muscle. It is currently available in the United States in oral and metered-dose inhaler forms. Nebulizer solutions and parenteral preparations are likely to be marketed here in the future. The chemical modifications that make albuterol beta 2-selective also promote oral bioavailability and increased duration of action by decreasing sensitivity to degradative enzymes. Albuterol can also produce undesirable dose-related effects: metabolic effects including decreased levels of plasma potassium, phosphate, calcium and magnesium; increased levels of plasma glucose, insulin, renin, lactate and ketones; peripheral vasodilation and perhaps some direct cardiac stimulation resulting in decreased systemic and pulmonary vascular resistance, increased pulse pressure and tachycardia; and skeletal muscle tremor. These side effects are most common with parenteral administration and much less prominent with aerosol administration, which yields lower systemic concentrations. Limited pharmacokinetic data suggest a long distribution phase, a terminal half-life of 3-8 hours, and 10-20% oral bioavailability. Aerosolization of albuterol or a similar agent with a compressed-air nebulizer appears to be best first-line management of the patient with acute dyspneic asthma, but appropriate preparations for this kind of therapy are currently missing from the United States market. Intravenous albuterol has also been employed in acutely dyspneic patients, but produces more side effects than carefully administered intravenous theophylline, is impaired by lack of sufficient pharmacokinetic information to guide dosing, and is of uncertain efficacy in the asthmatic with respiratory failure. However, it appears to lack the potentially life-threatening side effects that can result when theophylline is used carelessly . In the ambulatory patient, aerosolized albuterol (or a similar agent) administered by metered-dose inhaler is an excellent agent for treatment as needed and/or for prevention of acute bronchospasm triggered by exercise or other predictable cause. Advantages include a high degree of efficacy, rapid onset and long duration of effect, and minimal side effects. Regularly scheduled administration of albuterol by metered-dose inhaler is a widely used and effective maintenance medication for patients requiring long-term prophylactic therapy. However comparisons of the ability of this regimen and the other common maintenance regimens (cromolyn and theophylline) to control chronic symptoms of asthma are needed.
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PMID:Albuterol: an adrenergic agent for use in the treatment of asthma pharmacology, pharmacokinetics and clinical use. 673 11


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