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Efficacy of chlorhexidine solution with fluoride varnishing in preventing enamel softening by Streptococcus mutans in an artificial mouth.

For study of the enamel-protective effect of chlorhexidine-fluoride applications, the labial surfaces of pieces of bovine incisors were treated with 0.2% chlorhexidine gluconate solution, with Duraphat fluoride varnish, or with both of the above agents, while one group was treated with distilled water and one was left as an untreated control. Furthermore, a placebo varnish was used in the chlorhexidine- and distilled-water-treated groups; all the varnishes were removed after 24 h. The enamel slabs were mounted pairwise in an artificial mouth to form approximal contacts. The teeth were continuously rinsed with a common pool of artificial saliva to which was added 3% sucrose, and which was infected on the first day with Streptococcus mutans, "Ingbritt". The saliva was renewed daily and the incubation at 37 degrees C lasted for 10 days. The appreciable softening found in the distilled-water- and placebo-varnish-treated group tended to be prevented by the chlorhexidine and even more by the fluoride treatment, while the chlorhexidine-fluoride treatment prevented enamel softening completely. The saliva, infected only on the first day, and renewed daily, tended to become more acidified toward the end of the experimental period, obviously because the fermenting organisms had infected the surfaces of the model and formed plaque-like coatings on the enamel.

Animals↗

Changes in muscarinic ligand binding to intestinal muscle strips produced by pre-exposure to hypotonic conditions.

The extent of the binding of [3H]propylbenzilylcholine mustard (3H-PrBCM) to muscarinic receptors in longitudinal muscle strips from guinea-pig small intestine is increased by nearly 50% when the strips are preexposed to distilled water before measurement of 3H-PrBCM binding in Krebs-Henseleit solution. The apparent rate constant for 3H-PrBCM-receptor complex formation is more than double that of intact strips. The curves for the inhibition of 3H-PrBCM binding by methylatropinium bromide in normal and treated strips are superimposable, but, in contrast, distilled water pretreatment shifts the inhibition curve for carbachol to lower concentrations by a factor of 5-6. The inhibition curve for methylfurmethide is also shifted, by a factor of approximately 4, but the effect on the curve for hexyltrimethylammonium (C6TMA) is slight. The relative inhibition produced by benzhexol in the two preparations was variable. Comparison of the rate of equilibration of benzhexol with muscarinic receptors in intact and in distilled water pretreated muscle indicates that this inconsistency is unlikely to be due to incomplete equilibration.

Animals↗

Water uptake by Bufo melanostictus, as affected by osmotic gradients, vasopressin and temperature.

1. The rate of water uptake across the skin was studied in the live toad, Bufo melanostictus. When toads were kept in distilled water at 29 degrees C the uptake of water amounted to 16.9 +/- 1.3 mul./cm(2)/hr; when bathed in sucrose or urea solutions, the water uptake diminished with increasing osmotic pressure. There was no water uptake observed when toads were kept in 200 m-osmolar sucrose or urea.2. Intramuscular injections of vasopressin increased the rate of water uptake from distilled water. There was a good relation between doses and responses over various time intervals. A dose of 4 m-u. vasopressin/g body wt. doubled the rate of water uptake over a period of 1 hr. The same dose of vasopressin doubled the rate of water uptake when the toads were kept in solutions of sucrose or urea of different osmolarity.3. The rate of water uptake when the toads were bathed in sodium chloride solutions was consistently 8 mul./cm(2)/hr greater than when bathed in sucrose or urea solutions of equal osmolarity. There was no water uptake when the sodium chloride solution was 285 m-osmolar.4. Vasopressin (4 m-u./g) injected intramuscularly doubled the rate of water uptake from sodium chloride solutions of different osmolarity.5. With solutions of potassium chloride, sodium nitrate, and potassium nitrate, in concentrations up to 150 m-osmoles/l., the rate of water uptake was found to be the same as with solutions of sodium chloride of the same osmolarity. Similarly, it was doubled by injection of vasopressin (4m-u./g).6. The effect of temperature on the rate of water uptake before and after injection of vasopressin was investigated in toads kept in distilled water, sucrose, or sodium chloride solutions. For temperatures between 20 and 37 degrees C, vasopressin (4 m-u./g) reduced the activation energy involved in the process of water uptake by 4000 cal.7. The results agree with the view that water uptake follows a diffusion process which is facilitated by vasopressin, possibly as a result of increasing the size or number of available pores.

Animals↗

Water response of the frog olfactory epithelium as observed from the olfactory bulb.

The water response elicited by application of distilled water on the olfactory epithelium was recorded extracellularly from single olfactory bulb neurones. Characteristics of the water response in the frog olfactory epithelium were examined in comparison with those of the water response in the gustatory and palatal organs. 1. Effects of various electrolyte solutions on the generation of the water response were studied by dripping distilled water on the olfactory epithelium after adaptation to each of these electrolyte solutions. Number of the olfactory bulb cells responding to distilled water increased with increasing the charge of the adapting cations and also with decreasing the size of the cations with a few exceptions. 2. Magnitude of the 'water response' increased with decreasing concentration of salt in the solution which was dripped after adaptation to the isotonic solution of the same salt. 3. The water response was effectively depressed by an electrolyte solution but not by a non-electrolyte solution. An electrolyte also depressed effectively the water response which was produced after adaptation to an organic salt solution. 4. The water response was blocked by treatment of the olfactory epithelium with the uranyl ions which had high affinity for phospholipids. A tentative hypothesis on the generating mechanism of the water response in the frog olfactory epithelium is presented on the basis of the present experimental results and the water responses of the gustatory and palatal organs so far reported.

Action Potentials↗

Effect of amphotericin B dilution with various beverages on the survival of Candida albicans cells.

Amphotericin B (AMPH) has been generally used for prophylaxis or treatment of specific fungal diseases in immunocompromised patients. However, because it is difficult for children to ingest, mainly because of its bitter taste, it is often diluted with soft drinks. We therefore investigated the effect of dilution of AMPH with various beverages on its antifungal activity in vitro. Candida albicans cells were exposed for 30 min to AMPH diluted twofold with each of six commercially available beverages or distilled water, and percent survival was determined. The results showed 60% survival in the dilution with distilled water and higher survival when diluted with Yakult (136%; p < 0.01), orange juice (104%; p < 0.01), and coffee-milk (92%; p < 0.01). By contrast, lower survival was obtained when diluted with gum-syrup (54%), sweet cider (76%), and shaved-ice syrup (52%) with no significant differences from distilled water (60%), suggesting that these three beverages may be useful for diluting AMPH. The results of this study are a warning to medical workers that some methods of making AMPH more palatable considerably decrease its antifungal activity and may have a negative effect on host defenses against infectious diseases.

Amphotericin B↗

Cellulose Metabolism by the Termite Flagellate Trichomitopsis termopsidis.

The end products of cellulose metabolism by the trichomonad flagellate Trichomitopsis termopsidis from the termite Zootermopsis sp. were investigated by growing axenic flagellates on [C]cellulose. The growth of T. termopsidis resulted in the release of label into the supernatant fraction of the culture fluid, and > 75% was volatile under acid conditions. The label was analyzed for CO(2) and for [C]volatile compounds by vacuum distillation under acid and alkaline conditions in disposable micro-distillation vessels. The distillate and undistilled culture supernatant fluid were chromatographed on cellulose thin layers to identify the labeled end product. T. termopsidis produced CO(2) and [C]acetate which accounted for 25 to 30% and 55 to 60% of the labeled end products, respectively. The ratio of label in CO(2) to acetate suggests that they are produced in equimolar amounts. No neutral volatile compounds were produced. The remaining unidentified end product (10 to 20%) was not volatile nor extractable into ether. Hydrogen was produced by T. termopsidis, and the cells were killed by the drug metronidazole. Enzymatic activities were found which account for these end products: pyruvate:ferredoxin oxidoreductase and hydrogenase. The results indicate that acetate is the end product of T. termopsidis cellulose metabolism and is available to the termite for energy metabolism and biosynthesis.

Journal Article↗

Adsorption of reovirus to clay minerals: effects of cation-exchange capacity, cation saturation, and surface area.

The adsorption of reovirus to clay minerals has been reported by several investigators, but the mechanisms defining this association have been studied only minimally. The purpose of this investigation was to elucidate the mechanisms involved with this interaction. More reovirus type 3 was adsorbed, in both distilled and synthetic estuarine water, by low concentrations of montmorillonite than by comparable concentrations of kaolinite containing a mixed complement of cations on the exchange complex. Adsorption to the clays was essentially immediate and was correlated with the cation-exchange capacity of the clays, indicating that adsorption was primarily to negatively charged sites on the clays. Adsorption was greater with low concentrations of clays in estuarine water than in distilled water, as the higher ionic strength of the estuarine water reduced the electrokinetic potential of both clay and virus particles. The addition of cations (as chloride salts) to distilled water enhanced adsorption, with divalent cations being more effective than monovalent cations and 10(-2) M resulting in more adsorption than 10(-3) M. Potassium ions suppressed reovirus adsorption to montmorillonite, probably by collapsing the clay lattices and preventing the expression of the interlayer-derived cation-exchange capacity. More virus was adsorbed by montmorillonite made homoionic to various mono-, di-, and trivalent cations (except by montmorillonite homoionic to potassium) than by comparable concentrations of kaolinite homoionic to the same cations. The sequence of the amount of adsorption to homoionic montmorillonite was Al greater than Ca greater than Mg greater than Na greater than K; the sequence of adsorption to kaolinite was Na greater than Al greater than Ca greater than Mg greater than K. The constant partition-type adsorption isotherms obtained when the clay concentration was maintained constant and the virus concentration was varied indicated that a fixed proportion of the added virus population was adsorbed, regardless of the concentration of infectious particles. A heterogeneity within the reovirus population was indicated.

Adsorption↗

Effect of proteins on reovirus adsorption to clay minerals.

Organic matter in sewage, soil, and aquatic systems may enhance or inhibit the infectivity of viruses associated with particulates (e.g., clay minerals, sediments). The purpose of this investigation was to identify the mechanisms whereby organic matter, in the form of defined proteins, affects the adsorption of reovirus to the clay minerals kaolinite and montmorillonite and its subsequent infectivity. Chymotrypsin and ovalbumin reduced the adsorption of reovirus to kaolinite and montmorillonite homoionic to sodium. Lysozyme did not reduce the adsorption of the virus to kaolinite, but it did reduce adsorption to montmorillonite. The proteins apparently competed with the reovirus for sites on the clay. As lysozyme does not adsorb to kaolinite by cation exchange, it did not inhibit the adsorption of reovirus to this clay. The amount of reovirus desorbed from lysozyme-coated montmorillonite was approximately 38% less (compared with the input population) than that from uncoated or chymotrypsin-coated montmorillonite after six washings with sterile distilled water. Chymotrypsin and lysozyme markedly decreased reovirus infectivity in distilled water, whereas infectivity of the virus was enhanced after recovery from an ovalbumin-distilled water-reovirus suspension (i.e., from the immiscible pelleted fraction plus supernatant). The results of these studies indicate that the persistence of reovirus in terrestrial and aquatic environments may vary with the type of organic matter and clay mineral with which the virus comes in contact.

Adsorption↗

Membrane damage and microbial inactivation by chlorine in the absence and presence of a chlorine-demanding substrate.

The relationship between cell inactivation and membrane damage was studied in two gram-positive organisms, Listeria monocytogenes and Bacillus subtilis, and two gram-negative organisms, Yersinia enterocolitica and Escherichia coli, exposed to chlorine in the absence and presence of 150 ppm of organic matter (Trypticase soy broth). L. monocytogenes and B. subtilis were more resistant to chlorine in distilled water. The addition of small amounts of organic matter to the chlorination medium drastically increased the resistance of both types of microorganisms, but this effect was more marked in Y. enterocolitica and E. coli. In addition, the survival curves for these microorganisms in the presence of organic matter had a prolonged shoulder. Sublethal injury was not detected under most experimental conditions, and only gram-positive cells treated in distilled water showed a relevant degree of injury. The exposure of bacterial cells to chlorine in distilled water caused extensive permeabilization of the cytoplasmic membrane, but the concentrations required were much higher than those needed to inactivate cells. Therefore, there was no relationship between the occurrence of membrane permeabilization and cell death. The addition of organic matter to the treatment medium stabilized the cytoplasmic membrane against permeabilization in both the gram-positive and gram-negative bacteria investigated. Exposure of E. coli cells to the outer membrane-permeabilizing agent EDTA increased their sensitivity to chlorine and caused the shoulders in the survival curves to disappear. Based on these observations, we propose that bacterial envelopes could play a role in cell inactivation by modulating the access of chlorine to the key targets within the cell.

Cell Membrane↗

Identification of the predominant volatile compounds produced by Aspergillus flavus.

A culture of Aspergillus flavus grown on moistened wheat meal was homogenized with a blendor, and the resulting slurry was vacuum-distilled at 5 mm of Hg and 35 C. The aqueous distillate was collected in traps cooled to -10 to -80 C. The culture volatiles were extracted from the distillate with CH(2)Cl(2), and, after removal of the bulk of the solvent, the concentrated volatiles were examined by packed-column gas chromatography. Nineteen peaks were observed, and coupled gas chromatography-mass spectrometry was employed to identify the larger components. The compounds identified were: 3-methyl-butanol, 3-octanone, 3-octanol, 1-octen-3-ol, 1-octanol, and cis-2-octen-1-ol. The two octenols were the predominant compounds, and sufficient sample was trapped from the gas chromatograph for infrared analyses; this confirmed the mass spectral identifications and permitted the assignment of the cis designation to 2-octen-1-ol. Both oct-1-en-3-ol and cis-2-octen-1-ol are thought to be responsible for the characteristic musty-fungal odor of certain fungi; the latter compound may be a useful chemical index of fungal growth.

Aspergillus↗

Interaction of Mg-2+ with peptidoglycan and its relation to the prevention of lysis of a marine pseudomonad.

Intact cells of marine pseudomonad B-16 (ATCC 19855) which have been washed with a solution of NaCl require only 0.001 M MgSO4 and 100 to 300 times this concentration of NaCl or KCl to prevent lysis. Conversion of intact cells to mureinoplasts, a process involving removal of the outer double-track layer (outer membrane) and the periplasmic space layer of the cell wall, approximately doubled the requirement for the three salts to prevent lysis. The formation of protoplasts from mureinoplasts by removing the peptidoglycan layer again doubled the requirement for Na+ and K+ salts but increased the requirement for the Mg-2+ salt 200- to 300-fold. Cells of the marine pseudomonad suspended in solutions containing Mg-2+ salts failed to lyse on subsequent repeated suspension in distilled water, whereas cells presuspended in NaCl lysed immediately. Isolated envelope layers including the peptidoglycan layer, when dialyzed against solutiions containing Mg-2+ salts, retained Mg-2+ after subsequent suspension in distilled water. Envelope layers exposed to solutions of Na+ or K+ salts failed to retain these ions after exposure to distilled water. Na+ displaced Mg-2+ from the cell envelope layers. The results obtained indicate that the capacity of Mg-2+ salts at very low concentration to prevent lysis of intact cells and mureinoplasts of this organism is due primarily to the interaction of Mg-2+ with the peptidoglycan layer of the cell wall. Ion interaction with the layers lying outside of the peptidoglycan layer contributes only a small amount to the mechanical strength of the wall.

Bacteriolysis↗

Recovery of competence in calcium-limited Azotobacter vinelandii.

Azotobacter vinelandii cells required 0.5 mM calcium in the iron-limited competence induction medium. This requirement also was fulfilled by strontium, but not by magnesium. Cells pregrown in competence medium lacking calcium rapidly recovered competence with the addition of 0.5 mM calcium, provided they were suspended in the growth supernatant. A 60,000-dalton glycoprotein (pI 5.10) present in competent or incompetent culture supernatants participated in calcium-mediated competence recovery. Cells grown in calcium-limited medium appeared to have leaky cell envelopes and released a diverse array of proteins into the culture supernatant and into distilled water washes of the cells, seven of which appeared to be more dominant in competent cells. Two distilled water washes of cells grown in calcium-limited medium did not prevent calcium-mediated recovery of competence in the culture supernatant. Four to six distilled water washes removed a competence-specific protein (pI 5.19) and prevented calcium-mediated recovery of competence in the culture supernatant.

Azotobacter↗

Cation interactions and biochemical composition of the cell envelope of a marine bacterium.

Envelopes of a marine isolate, c-A1, and of a terrestrial isolate, 121, were compared for their susceptibility to disintegration in distilled water after exposure to 0.05 m MgCl(2) and to 0.1 and 1.0 m NaCl. After exposure to MgCl(2) alone, both types of envelopes remained intact in distilled water. Envelopes of marine isolate c-A1, but not of the terrestrial isolate, fragmented in distilled water after exposure to 1.0 m NaCl. Partial reaggregation of the c-A1 envelope fragments occurred on addition of MgCl(2). In cation-exchange experiments, bound Mg(++) in the envelopes of both organisms was displaced by Na(+). The envelopes of c-A1 were found to contain lipopolysaccharide, muramic acid, and a variety of phospholipids, of which the major component was phosphatidylethanolamine, accompanied by lesser amounts of phosphatidic acid, diphosphatidylglycerol, and phosphatidylserine. Analyses of envelope acid hydrolysates revealed a similar amino acid distribution in the marine and terrestrial isolates, but envelopes of c-A1 had less than half the total amino acid content of envelopes of 121 per envelope dry weight. Possible relationships between cations and biochemical components of the envelopes are considered in terms of differences in behavior of the two organisms in low ionic environments.

Amino Acids↗

Bronchial reactivity in asthmatic adults with normal spirometric values.

A comparison was made between airways obstruction produced by exercise. Hyperventilation of cold air, hyperventilation of room air, and inhalation of ultrasonically nebulised distilled water. Seven asthmatics with no airflow obstruction were studied together with seven normal controls. All four provocations produced comparable mean reductions in forced expired volume in one second in the asthmatic patients, but no reduction was obtained in the controls. Whereas exercise, hyperventilation of cold air, and hyperventilation of room air were associated with heat loss from airways, inhalation of nebulised distilled water was not. Inhalation of ultrasonically nebulised distilled water offers a simple additional diagnostic test for asthma. Changes in the osmolality of airway surface liquid may possibly be the common mechanism by which "fog" and hyperventilation produce bronchoconstruction.

Adult↗

Micromethodology for measuring ATPase activity in renal tubules: mineralocorticoid influence.

The ATPase activity of rabbit isolated renal tubule segments was measured using a microtechnique in which the hydrolysis of ATP was enzymatically coupled to the appearance of an alkali-converted, highly fluorescent form of nicotinamide adenine dinucleotide. The methods are simple, reproducible, and have a high sensitivity in which picomole quantities of hydrolyzed ATP can readily be measured. Several methods for permeabilizing the cell membranes for measurement of Na+-K+-ATPase activity were evaluated, including osmotic (distilled water or 300 mM imidazole) and temperature (freezing) shock and addition of the nonionic detergent octylglucoside. An octylglucoside concentration of 0.5% was found to cause a maximum activation of the Na+-K+-ATPase and was comparable with that observed when tubules were permeabilized by exposure to distilled water and freezing. Incubation of tubules in 300 mM imidazole was less effective in permeabilizing the cell membranes. In all subsequent studies, the cells were permeabilized by exposure to distilled water and freezing as done by others. The methods were used to assay for the basal levels of Na+-K+-ATPase in the superficial proximal convoluted tubule, the superficial proximal straight tubule, and the cortical collecting tubule and were found to average 44.9 +/- 6.3, 26.4 +/- 2.4, and 11.8 +/- 2.2 pmol ADP X mm-1 X min-1, respectively. Furthermore, elevation of plasma mineralocorticoids by daily injections of deoxycorticosterone acetate (2 mg X kg-1 X day-1) for 4-15 days caused a doubling in the Na+-K+-ATPase activity of the cortical collecting duct, confirming the results of others. The methods presented can easily be adapted for microanalysis of other ATPases.

Adenosine Diphosphate↗

Regulation of body volume by salivation in a tick challenged with fluid loads.

Injection into the hemolymph of 1.2% NaCl, 11.2% sucrose, 2.3% urea (all approximately isosmotic to hemolymph), or distilled water induced salivary fluid secretion in the ixodid tick Amblyomma hebraeum Koch. Saline gave the largest response at high doses. Injection of hyperosmotic NaCL into the hemolymph did not induce salivation but led to the drinking of distilled water in amounts sufficient to dilute the salt load to isosmolarity. Atropine only partially inhibited salivation induced by NaCl, sucrose, and distilled water. Reserpine markedly inhibited salivation induced by NaCl. We propose that at least two sensory pathways (one cholinergic, one not) converge on the secretory nerve. The physiological significance of the cholinergic pathway is not known. The other pathway probably mediates the regulation of hemolymph volume, possibly via stretch receptors, but its transmitter is not known.

Animals↗

Osmotic factors in restitution from thermal dehydration in rats.

The degree of voluntary dehydration after thermal dehydration was assessed while supplying drinking water of varying NaCl concentrations. Adult male albino rats were exposed to a hot-dry environment (dry bulb temp 36 degrees C; relative humidity 20%) for 6-8 h, and dehydration of 7% body wt was induced. After dehydration the rats were divided into three groups, and distilled water and 0.45 or 0.9% NaCl solution was given ad libitum. The amount of fluid intake reached an equilibrium in 6-12 h; the 0.45 and 0.9% NaCl groups regained almost all lost water, whereas the distilled water group regained only half. During rehydration the 0.45 and 0.9% NaCl groups gained Na by approximately 430-650 mueq and lost K by 90-130 mueq, whereas the distilled water group lost Na slightly and K by 80 mueq/100 g body wt. As for the electrolyte balance during thermal dehydration, rats excreted Na and K into urine and saliva. Na loss was 200 mueq/100 g, almost all of which was derived from the interstitial space of skin and skeletal muscle, and K loss was 230 mueq/100 g, almost all of which was derived from intracellular space of skeletal muscle. Total cation loss (Na, K) during thermal dehydration, including K excretion during rehydration, was 510-560 mueq/100 g, which was almost identical to the Na gained by rats given 0.45 or 0.9% NaCl solution. These results suggest that voluntary dehydration is caused by the dilutional inhibition of drinking due to loss of electrolytes during thermal dehydration.

Animals↗

Dopamine D1 receptor antagonist inhibits swallowing reflex in guinea pigs.

To determine whether dopamine D1 receptor antagonist impairs the swallowing reflex and reduces substance P (SP) in the peripheral organs, the swallowing reflex in terms of the number of swallows elicited by injections of three different volumes (0.2, 0.4, and 0.6 ml) of distilled water into the pharynx through a catheter was examined in anesthetized guinea pigs pretreated with Sch-23390. Animals were pretreated with either subcutaneous Sch-23390 (200 micrograms/kg) or a vehicle of Sch-23390 every 12 h for 7 days. The number of swallows was counted by submental electromyographic activity and visual observation of characteristic laryngeal movement. Injections of distilled water caused a volume-dependent increase in the number of swallows in animals without Sch-23390 treatment. Sch-23390 significantly decreased and exogenously administered SP increased the number of swallows elicited by all volumes of distilled water. FK-888 (10(-5) M, 1 ml), a specific inhibitor of the NK1 receptor, reduced the number of swallows to a greater degree than Sch-23390. Sch-23390 significantly reduced SP content in the laryngeal and pharyngeal mucosa compared with control. These results suggest that inhibition of the dopamine D1 receptor may impair the swallowing reflex and reduce SP content in the peripheral organs.

Animals↗