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Biomedical subjects

C F Schachtele

Publications and source records attributed to C F Schachtele.

At least 19 recordsLinked to original sources
PubMed1997

Acid tolerance and acid-neutralizing activity of Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum.

The tolerance to acid and the acid-neutralizing activity of three important periodontopathic bacteria, Porphyromonas gingivalis, Prevotella intermedia and Fusobacterium nucleatum were studied. P. gingivalis strains grew only at neutral pH and did not utilize glucose, whereas strains of P. intermedia and F. nucleatum could grow under acidic conditions and increased their growth by utilizing glucose. P. gingivalis tended to raise the culture pH during growth. P. intermedia and F. nucleatum raised the culture pH during growth in the absence of glucose, while in the presence of glucose they decreased the pH. Resting cell suspensions of all the bacteria raised the pH in the presence of tryptone and casamino acids. Acid-neutralizing activity was confirmed by measuring base production at a fixed pH with a pH-stat. During neutralization, the cells produced cytotoxic substrates, ammonia and organic acids (butyric, isobutyric and isovaleric acids by P. gingivalis; isovaleric and succinic acids by P. intermedia; propionic and butyric acids by F. nucleatum). These findings suggest that deamination of amino acids into ammonia and organic acids occurs simultaneously with base production, resulting in acid neutralization. These results could partially explain the survival of P. intermedia and F. nucleatum in both supragingival and subgingival plaque and the apparent restriction of P. gingivalis to subgingival plaque. The former bacteria may aid in creation of an environment fostering colonization of subgingival plaque by P. gingivalis.

Culture Media↗
PubMed1995

Simultaneous monitoring of intracellular pH and proton excretion during glycolysis by Streptococcus mutans and Streptococcus sanguis: effect of low pH and fluoride.

A system was developed by which 2',7'-bis(carboxyethyl)-4 or 5-carboxyfluorescein could be used to monitor intracellular pH at the same time that proton excretion was being measured. Streptococcal cells were loaded with the dye, and after the addition of glucose protons were excreted and the intracellular pH increased quickly and remained higher than the extracellular pH of 7.0. The excretion of protons stopped and the intracellular pH returned to the original level when glucose was depleted. The intracellular level of ATP remained high during glucose metabolism and decreased with the depletion of glucose. At extracellular pH of 5.5, and 5.0, the intracellular pH of fasting cells was higher than the extracellular pH value. After addition of glucose there were initial lags of proton excretion and of increases in intracellular pH at the acidic extracellular pH values. In the presence of fluoride, a lag in proton excretion and a simultaneous decrease in intracellular pH were observed, indicating a partial and transient inhibition of proton-ATPase activity.

Adenosine Triphosphate↗
PubMed1995

Mechanism of inhibition of glycolysis in Streptococcus mutans NCIB 11723 by chlorhexidine.

Inhibition of the rate of acid production from glucose by the cells of Streptococcus mutans NCIB 11723 was directly related to the concentrations of 0.075 to 0.20 mM chlorhexidine. Lactate production was inhibited to a greater extent than acetate and formate. Quantification of glycolytic intermediates revealed that the steps in glycolysis inhibited by chlorhexidine were the reactions catalyzed by phosphofructokinase and glyceraldehyde 3-phosphate dehydrogenase and/or phosphoglycerate kinase. However, the activities of these enzymes were not decreased in cells treated with the inhibitor. It was demonstrated that chlorhexidine caused leakage of metabolites from the cells. Our results indicate that the decreased rate of glycolysis caused by chlorhexidine is due to the leakage of metabolic intermediates and not to direct effects on enzymes involved in glycolysis by S. mutans NCIB 11723.

Acetates↗
PubMed1995

Effect of sucrose monolaurate on acid production, levels of glycolytic intermediates, and enzyme activities of Streptococcus mutans NCTC 10449.

We studied the mechanism by which the antimicrobial compound sucrose monolaurate inhibits Streptococcus mutans NCTC 10449 by determining its effect on the rate of acid production from glucose and sucrose and the intracellular and extracellular levels of glycolytic intermediates. Sucrose monolaurate was more effective than either sodium laurate or sodium fluoride in inhibiting acid production at pH 7.0 from glucose. Inhibition of acid production was the same when either glucose or sucrose was the carbon source and in the presence or absence of oxygen. Quantitative analysis of various glycolytic intermediates revealed that the steps inhibited by sucrose monolaurate were the reactions catalyzed by phosphofructokinase and glyceraldehyde 3-phosphate dehydrogenase and/or phosphoglycerate kinase. Since the activities of these enzymes in cell-free extracts were not decreased by the addition of sucrose monolaurate, the inhibition of acid production could not be ascribed to direct effects on the enzymes. A decrease in the rate of acid production with corresponding elevations in the extracellular levels of glycolytic intermediates indicates that sucrose monolaurate inhibits S. mutans by altering the permeability of the cell membrane, which causes a loss of important metabolites.

Anti-Infective Agents, Local↗
PubMed1992

Effect of gum chewing following food ingestion on the pH of interproximal dental plaque.

Recent publications have suggested that chewing sorbitol- or sucrose-containing gum after a snack or meal can reduce development of caries by neutralizing dental plaque acids at interproximal sites in the dentition. To confirm these findings four volunteers wore appliances containing a miniature pH electrode. After plaque accumulation, subjects ingested a bowl of sugar-coated cereal with milk and 20 minutes later chewed a sorbitol-containing gum, a sucrose-containing gum, or did not chew anything for 20 minutes. After exposure to the cereal, the plaque pH fell within 20 minutes from approximately 6.4 to 4.8. Sorbitol gum caused the pH to rise to 5.5, while the sucrose gum caused the pH to rise to only 5.1. After cessation of chewing, the pH in all cases dropped to 4.5 or lower. No statistically significant difference could be shown between plaque pH changes with the various protocols. Gum chewing after eating caused only a transient elevation in plaque pH.

Adult↗
PubMed1990

Effect of dental plaque age and bacterial composition on the pH of artificial fissures in human volunteers.

Changes in sucrose-induced plaque pH profiles and the microbial composition of occlusal tooth surface fissures were analyzed using wire telemetry and bacterial culturing techniques. Four human volunteers wore appliances containing artificial fissures constructed with ion-sensitive field-effect transistor (ISFET) electrodes for 1, 2 and 4 days; 1 subject kept the electrode for 3 weeks. After monitoring the plaque pH response at the base of the fissure to a 10% (w/v) sucrose rinse the plaque was removed and analyzed for total viable bacteria, total and specific streptococci, lactobacilli and Actinomyces spp. One-day-old plaque showed a rapid drop in plaque pH to a minimum of 4.8 +/- 0.2, with 2-day-old plaque showing the most acidogenic pH profile (minimum pH 4.6 +/- 0.2). The 4-day-old plaque response was less acidogenic (minimum pH 5.0 +/- 0.3) than the results from days 1 and 2. Responses from 13- and 21-day-old fissure plaques showed greatly decreased acidogenic responses (day 21 minimum pH 5.7). Viable bacteria recovered from the fissure increased from approximately 4 x 10(6) colony-forming units on day 1 to 1.2 x 10(7) on days 2 and 4 and 1.7 x 10(7) on day 21. Streptococci (greater than 50%) and Actinomyces (greater than 10%) dominated in the fissure plaques and their levels were related to minimum pH. Since fissure plaque of all ages tested contained high concentrations of acidogenic bacteria, the decreased acidogenic response at the base of fissures with increasing plaque age suggests that maturing fissure plaques provide an increasingly greater diffusion barrier to fermentable carbohydrates.

Actinomyces↗
PubMed1990

Effect of pH on the growth and proteolytic activity of Porphyromonas gingivalis and Bacteroides intermedius.

The effect of pH on the growth and proteolytic activity of the type strain and fresh isolates of Porphyromonas gingivalis and Bacteroides intermedius was investigated. B. intermedius strains grew with and without glucose at a pH as low as 5.0. These bacteria grew almost as well as Streptococcus mutans at pH 5.0 and better than Actinomyces viscosus at pH 5.5 and 5.0. Some B. intermedius strains raised the culture pH when grown at a low pH without glucose. In contrast, P. gingivalis strains grew only at pH 6.5 to 7.0. The P. gingivalis strains had proteolytic activities against azocoll, azocasein, and azoalbumin, while the B. intermedius strains degraded azocasein and azoalbumin, but not azocoll. B. intermedius showed maximum proteolytic activity at pH 7.0, and high activity over a wide pH range. In contrast, the optimum pH of proteolytic activity in P. gingivalis was pH 7.5 to 8.0. The P. gingivalis activities were more sensitive than those of B. intermedius to low pH. The capacity of B. intermedius to degrade proteins to more readily metabolizable substrates at low pH might explain the growth of this bacterium in an acidic environment. These differences between B. intermedius and P. gingivalis could explain their capacity to survive at different sites in the oral cavity and indicate how B. intermedius might positively influence the growth of P. gingivalis in subgingival plaque.

Albumins↗
PubMed1989

Comparison of in vivo human dental plaque pH changes within artificial fissures and at interproximal sites.

Ion-sensitive field-effect transistor (ISFET) pH electrodes were used to monitor changes in plaque pH at the base of artificial occlusal surface fissures and at interproximal sites. Bovine enamel was used to construct fissures (1.5 x 0.1 x 1.0 mm) containing a small ISFET electrode. The fissures were fixed to carrier appliances and worn by 4 human volunteers. After plaque accumulation for 4 days changes in pH were monitored by wire telemetry following 1-min rinses with 10% solutions of either sorbitol or sucrose. Results were compared to data obtained from interproximal sites in the same subjects. Responses to sorbitol in the fissure and on the proximal surfaces were minimal and showed no significant difference in minimum pH (5.9 +/- 0.4 and 6.1 +/- 0.3, respectively) and area under pH 7.0. The response to sucrose at the two sites was very different revealing unique pH profiles which were statistically significantly different with regard to minimum pH (5.0 +/- 0.3, fissure and 4.3 +/- 0.2, proximal) and area under pH 5.7. Thus, the acidogenic potential of fermentable carbohydrate at two caries-prone sites in the human dentition is significantly different and conclusions based on interproximal telemetry measurements may not be applicable to occlusal surface fissures.

Acid-Base Equilibrium↗
PubMed1988

Effect of chewing gum containing sodium bicarbonate on human interproximal plaque pH.

The effect of chewing sorbitol gum containing sodium bicarbonate on interproximal plaque pH was determined by use of an ion-sensitive field-effect transistor (ISFET) electrode system with four human volunteers. Plaque pH was lowered by 2 min of chewing of a sucrose-containing toffee. After 20 min, either sodium bicarbonate or control gum was chewed for 10 min, and the pH was followed for an additional 20 min. The minimum pH after toffee chewing was elevated by both the control gum (4.5 +/- 0.3 to 5.2 +/- 0.5) and the sodium-bicarbonate-containing product (4.3 +/- 0.3 to 6.1 +/- 0.6) to levels which were significantly different (p less than 0.01). The rate of rise in pH was 2.6 times faster with the gum containing buffer compared with the control gum. In comparison with the control gum, the sodium bicarbonate gum caused the pH to remain at a higher level, approximately 0.5 pH units, for 20 min after gum chewing. The addition of sodium bicarbonate to gum containing sorbitol markedly enhanced its capacity to cause and maintain an elevation of interproximal plaque pH previously lowered by exposure to fermentable carbohydrate. Sodium bicarbonate may be useful in products designed to reduce the acidogenic challenge to the teeth following food ingestion.

Adult↗
PubMed1988

Utilization of a continuous streptococcal surface to measure interbacterial adherence in vitro and in vivo.

Cell-to-cell interactions are essential for the formation of dental plaque. A continuous layer of Streptococcus sanguis SA-1 cells fixed to a solid surface has been used to evaluate interactions among this bacterium, Haemophilus parainfluenzae, and Streptococcus sobrinus. S. sanguis cells were attached to a Falcon 3001 tissue culture plates or bovine enamel chips, coated with a biological adhesive. Scanning electron microscopy of the chips showed the streptococci as a contiguous surface. Radiolabeled bacteria were used to measure a second-species interbacterial adherence to the streptococcal-coated culture plates. Strains of H. parainfluenzae known to coaggregate (strain HP-28) and not to coaggregate (strains HP-42 and HP-80), in suspension with S. sanguis strain SA-1, were studied for adherence. Ten-fold-higher numbers of coaggregating strain HP-28 adhered in vitro to the streptococcal layer than did the non-coaggregating strains. S. sobrinus strain 6715 did not show appreciable adherence to the S. sanguis surface. Saliva did not affect the adherence of coaggregating or non-coaggregating H. parainfluenzae strains to S. sanguis strain SA-1. Bovine enamel chips, coated with streptococci, mounted on modified orthodontic appliances and placed in the mouths of three volunteers, facilitated the measurement of interbacterial adherence in vivo of streptomycin-resistant strains of H. parainfluenzae (HP-28R or HP-42R). Suspensions of bacteria were placed into the mouth, distributed throughout, and expectorated. After 15 or 120 minutes, the appliance with the chips was removed, the chips sonified, and colony-forming units (CFU) of streptomycin-resistant haemophili determined per chip.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

Molecular cloning of the lactose-metabolizing genes from Streptococcus lactis.

Restriction endonucleases and agarose gel electrophoresis were used to analyze plasmid pLM2001, which is required for lactose metabolism by Streptococcus lactis LM0232. The enzymes XhoI, SstI, BamHI, and KpnI each cleaved the plasmid into two fragments, whereas EcoRI and BglII cleaved the plasmid into seven and five fragments, respectively. Sizing of fragments and multiple digestions allowed construction of a composite restriction map. The KpnI fragments of pLM2001 were cloned into the KpnI cleavage site of the vector plasmid pDB101. A recombinant plasmid (pSH3) obtained from a lactose-fermenting, erythromycin-resistant (Lac+ Eryr) transformant of Streptococcus sanguis Challis was analyzed by enzyme digestion and agarose gel electrophoresis. Plasmid pSH3 contained 7 of the 11 KpnI-HindIII fragments from pLM2001 and 5 of the 7 fragments from pDB101. It was determined that a 23-kilobase (kb) KpnI-generated fragment from pLM2001 had been cloned into pDB101 with deletion of part of the vector plasmid. The recombinant plasmid could be transformed with high frequency into several Lac- strains of S. sanguis, conferring the ability to ferment lactose and erythromycin resistance. The presence of pSH3 allowed a strain deficient in Enzyme IIlac, Factor IIIlac, and phospho-beta-galactosidase of the lactose phosphoenolpyruvate-dependent phosphotransferase system to efficiently ferment lactose. Under conditions designed to maximize curing of plasmid DNA with acriflavin, no Lac- derivatives could be isolated from cells transformed with pSH3. Seven of the 40 Lac+ colonies isolated after 10 transfers in acriflavin were shown to be sensitive to erythromycin and did not appear to harbor plasmid DNA.(ABSTRACT TRUNCATED AT 250 WORDS)

Base Sequence↗
PubMed1984

The effect of an antibacterial solution on the microflora of human incipient fissure caries.

A technique for collecting various levels of plaque from incipient occlusal fissure caries was used to evaluate the antibacterial properties of a solution containing I2-KI, CuSO4, and ethanol. In comparison with control fissures (n = 6), the treated fissures (n = 9) showed a 98% difference in recoverable bacteria, with the killing of S. mutans, Actinomyces, and lactobacilli appearing to reflect the level of agent penetration.

Actinomyces↗
PubMed1984

Fissure removal and needle scraping for evaluation of the bacteria in occlusal fissures of human teeth.

The bacterial counts obtained with a needle-scrape method for collecting plaque samples from human occlusal fissures with incipient caries were compared with the microflora remaining in the fissures as determined with a fissure removal method. Scraping of six fissures with a sterile needle recovered only 18.2% of the total recovered fissure flora. The needle-scrape method failed to detect specific cariogenic bacteria which were present in four of the fissures. Treatment of nine fissures with an antibacterial solution demonstrated that the needle method failed to detect viable bacteria in eight of the fissures which were subsequently shown to contain bacteria by fissure removal. The commonly used needle-scraping method does not appear adequate for studies on the effect of antimicrobial agents on fissure plaque.

Adolescent↗
PubMed1983

The acidogenic potential of reference foods and snacks at interproximal sites in the human dentition.

Telemetry was used to evaluate changes in plaque pH at interproximal sites in the dentition of five human subjects following the ingestion of a variety of reference foods and snacks. Short-term telemetry (30 min) revealed that most of the substrates yielded both pH minima and total responses similar to that obtained with a 10% sucrose rinse. Aged Cheddar cheese and skim milk were much less acidogenic than were the other foods.

Acids↗
PubMed1983

Plaque pH measurements by different methods on the buccal and approximal surfaces of human teeth after a sucrose rinse.

Changes in plaque pH after a sucrose rinse were simultaneously monitored by plaque sampling, touching of the plaque with antimony and glass electrodes, and telemetry. The minimum pH at approximal sites was approximately 0.7 pH units lower than that on buccal surfaces. The pH at the approximal site remained below resting levels after 120 min, and the area under the pH response curves from this site was five times greater than that from the buccal surfaces.

Dental Plaque↗
PubMed1982

Plaque sampling and telemetry for monitoring acid production on human buccal tooth surfaces.

The plaque sampling method and wire telemetry using miniature, glass pH electrodes on the buccal surface of mandibular first permanent molars, compared human dental plaque pH responses to a 10 per cent solution of sucrose. Nine subjects abstained from oral hygiene and were tested in morning sessions on 5 consecutive days. The mean resting plaque pH value for all sessions by plaque sampling was 6.60 +/- 0.18 (mean +/- SD) and by telemetry was 6.56 +/- 0.19. After a 1 min sucrose rinse, 1-day-old plaque showed a decrease in pH approx. 5.5 by both methods. The 2-, 3-, 4- and 5-day-old plaque, the mean minimum pH achieved was 4.69 +/- 0.28 as measured by sampling and 3.94 +/- 0.49 as determined by telemetry. The times taken to reach minimum pH as determined by the two methods were almost identical. The electrode calibration data showed that all the telemetry electrodes responded consistently for all 5 days of study. Scanning electron microscopy and microbiological analysis of electrode tips and enamel replicas revealed that plaque accumulated on glass at the same rate with a similar bacterial composition to plaque formed on clean enamel. It is proposed that glass microelectrodes and wire telemetry are a reasonable means for continuously monitoring plaque pH in situ. Comparison with plaque sampling suggested that telemetric responses reflect the type of plaque which accumulates at particular sites on the dentition.

Adult↗