The relation of the tricarboxylic acid cycle to bacterial infection. IV. The effect of three metabolic inhibitors and Salmonella typhimurium on the citric acid content of mouse tissues.
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The ability of eight strains of Aspergillus niger to produce citric acid by the solid surface method were found to correlate with their capabilities to synthesize intracellular enzymes which degrade phytates (phytase and acid phosphatase). Another high correlation was observed between phytase and acid phosphatase activities bound to the cell walls of mycelia.
In rats of three age groups (one month-old rattlings, young 3-month old and adult rats not younger than 5 months) receiving the usual vivarium food the exhalation of C14O2 substantially decreased with the age and the maximum radioactivity of the air following intraperitoneal introduction of citric acid-3-C14 was greatly delayed. Upon putting for 2 months on a cariogenic diet with 54 per cent of saccharose of one month old and 3-month old animals there was noted a significantly accelerated appearance in the expired air of C14O2 and a quicker reaching the maximum radioactivity of the expired air than in controls. It seems that in an accelerated exhalation of C14O2 a definite role is played by the transormation of citric acid in the muscles and blood with the animals kept on a saccharose diet.
The citric acid or tricarboxylic acid cycle is a central element of higher-plant carbon metabolism which provides, among other things, electrons for oxidative phosphorylation in the inner mitochondrial membrane, intermediates for amino-acid biosynthesis, and oxaloacetate for gluconeogenesis from succinate derived from fatty acids via the glyoxylate cycle in glyoxysomes. The tricarboxylic acid cycle is a typical mitochondrial pathway and is widespread among alpha-proteobacteria, the group of eubacteria as defined under rRNA systematics from which mitochondria arose. Most of the enzymes of the tricarboxylic acid cycle are encoded in the nucleus in higher eukaryotes, and several have been previously shown to branch with their homologues from alpha-proteobacteria, indicating that the eukaryotic nuclear genes were acquired from the mitochondrial genome during the course of evolution. Here, we investigate the individual evolutionary histories of all of the enzymes of the tricarboxylic acid cycle and the glyoxylate cycle using protein maximum likelihood phylogenies, focusing on the evolutionary origin of the nuclear-encoded proteins in higher plants. The results indicate that about half of the proteins involved in this eukaryotic pathway are most similar to their alpha-proteobacterial homologues, whereas the remainder are most similar to eubacterial, but not specifically alpha-proteobacterial, homologues. A consideration of (a) the process of lateral gene transfer among free-living prokaryotes and (b) the mechanistics of endosymbiotic (symbiont-to-host) gene transfer reveals that it is unrealistic to expect all nuclear genes that were acquired from the alpha-proteobacterial ancestor of mitochondria to branch specifically with their homologues encoded in the genomes of contemporary alpha-proteobacteria. Rather, even if molecular phylogenetics were to work perfectly (which it does not), then some nuclear-encoded proteins that were acquired from the alpha-proteobacterial ancestor of mitochondria should, in phylogenetic trees, branch with homologues that are no longer found in most alpha-proteobacterial genomes, and some should reside on long branches that reveal affinity to eubacterial rather than archaebacterial homologues, but no particular affinity for any specific eubacterial donor.
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Acid-treatment of root surfaces as adjunct to periodontal reconstructive surgery may have deleterious effects on that part which remains in a supragingival location after healing. The present experiment aimed to examine whether recalcification of such root surfaces may occur. Cylindrical blocks of root dentin were prepared for 10 premolars, treated with citric acid and mounted in acrylic specimen holders which were worn in the mouth continuously for 14 d by two test subjects. The specimens were then sectioned and microradiographs produced. The citric acid had produced a completely demineralized surface zone in the root dentin and the subjacent hard tissue showed a zone of partially reduced mineral content. Following exposure to the oral cavity, the demineralized surface layer appeared unchanged in width. The subjacent zone, however, had been reduced in width in all instances when compared with control specimens from the same teeth.
Five extracted human teeth with clinically demonstrable calculus were used. One proximal surface of each tooth was partly scaled, leaving a small amount of calculus. The crown was then cut off at the cementoenamel junction and the scaled proximal surface was divided vertically into two segments using a thin separating disk. One segment was treated by rubbing cotton pellets soaked in citric acid (pH1) over the surface for three minutes and then immersing the segment in saline to stop the reaction. The other (control) segment was treated by rubbing cotton pellets soaked in saline over the surface for three minutes. When viewed under a scanning electron microscope, the control sections displayed scaling striations, considerable surface debris, and large numbers of bacteria at the borders of the residual calculus. Citric acid-treated specimens displayed little debris on cementum or residual calculus and virtually no bacteria at the junction between calculus and cementum. The surface morphology of the citric acid-treated calculus varied from layered-like to honeycomb.
This study compared the antimicrobial effect of MTAD, two of its components, doxycycline and citric acid, and sodium hypochlorite (NaOCl) in two in vitro models on Enterococcus faecalis. In the bovine tooth model, the lumens of 30 bovine dentin discs were infected with E. faecalis for 2 weeks before treating with either one of the experimental irrigants or saline. Bacteria in the shavings were collected with two sizes of burs and enumerated after overnight culturing. Zones of inhibition were recorded in the agar diffusion model for each irrigant. In the tooth model, NaOCl and doxycycline were more effective than control in killing E. faecalis at the shallow bur depth, but at the deeper bur depth only NaOCl was superior. In the agar diffusion model, NaOCl produced less inhibition than MTAD or doxycycline.
The changes in the rates of citrate biosynthesis and utilization in rat brain, liver, kidney and heart, produced by hypoxia, action of 2,4-DNP and thyreotoxicosis, were compared with changes of some regulatory parameters under the same conditions. The comparison of citrate-synthase activities, citrate levels in tissues and 14C-incorporation from different precursors into citric acid permitted us to establish that the biosynthesis of citrate in brain was more intensive than in other tissues studied. The main source of acetyl-CoA for citrate-synthase reaction in brain is the oxidation of pyruvate. The ratio of adenine nucleotides plays an important role in the control of citrate-synthase activity in brain, where the oxaloacetate control is not as significant as in liver. NAD-specific isocitrate dehydrogenase reaction was found to be the dominant pathway for citrate oxidation in brain: more than 60 percent of brain citrate were oxidized by NAD-ICDH, while less than 10 percent of citric acid were utilized by this enzyme in other tissues studied. The existance of an adenine nucleotide control of NAD-ICDH activity in brain may be an additional mechanism for the regulation of the first steps of energy metabolism in brain.
The effect of dietary citric acid (CA) and fumaric acid (FA) on pig weight gain (ADG) and gain/feed (G/F) was studied in two trials using 192 crossbred, 4-wk-old weaning pigs. Three dietary levels (0, 1.5 or 3.0%) of either FA (Trial 1) or CA (Trial 2) with or without an antibiotic supplement (110 mg chlortetracycline, 110 mg sulfamethazine and 55 mg penicillin/kg diet) formed six treatment combinations in each trial. These six diets were fed to two replicate pens of eight pigs each for a 4-wk period. In Trial 1, ADG was improved (P less than .01) during wk 1, and G/F was improved during wk 1 (P less than .01) as well as during wk 1 to 2 (P less than .05) for pigs consuming FA-supplemented diets. In Trial 2, CA had no beneficial influence on ADG during the 4-wk trial. However, feed intake during wk 1 was depressed (P less than .05) by adding CA, as was G/F during wk 1 to 2 (P less than .05). Based on these results, FA was selected to be used in a nutrient balance study. Twelve 4-wk-old weanling pigs were fed one of three diets: control (C), C + 1.5% FA, or C + antibiotic supplement (A). Diet digestible energy (DE), ME and N-corrected ME (MEN) were not different among treatments. Nitrogen balance, percentage N retained and apparent N digestibility were not affected by dietary treatment. Calcium balance and percentage of Ca retained were unaffected by diet.(ABSTRACT TRUNCATED AT 250 WORDS)
Rats with permanent access to a water supply adulterated with citric acid (CA) displayed persistent reductions in fluid intake and fluid/food ratios; and at appropriate concentrations of CA, they also exhibited lowered body weights and drinking deficits after fluid deprivation and after hypertonic NaCl injections. Unlike rats in previous investigations that were forced to consume quinine adulterated water, CA drinkers exhibited less disruption in their ingestive behavior following regulatory challenges, diminished short-term but greater long-term abilities in responding to hypovolemia, and an ability to increase fluid intake after fluid deprivation plus hypertonic NaCl. These data reveal that substances which degrade the taste quality of water do not exert a unitary influence on fluid intake, and they further underscore the complexity of ecological factors involved in controlling drinking behavior.
Improvement of new attachment procedures using topically applied citric acid was tested in human beings. Furcation involvement and the adjacent proximal area were surgically exposed. Saline was used as placebo drug. Baseline pocket depth and attachment levels were scored prior to surgery and bone level recorded at the time of surgery. After 1, 3 and 6 months, the clinical parameters were re-scored and at that time, a re-entry operation allowed a new measurement of the bone level. In the post-operative experimental periods, there was an improvement of all clinical parameters in both the control and experimental groups. Pocket depth reduction was consistent; the gain in the attachment level compared to the baseline examination in the experimental group and the improvement in the bone height level were statistically significant within the 6-month post-operative interval; however, in both the furcation and proximal areas, no statistically significant differences could be detected on comparing experimental and control groups.
This paper reviews the operation of a full-scale, fixed-bed digester treating a citric acid production wastewater with a COD:sulphate ratio of 3-4:1. Support matrix pieces were removed from the digester at intervals during the first 5 years of operation in order to quantify the vertical distribution of biomass within the digester. Detailed analysis of the digester biomass after 5 years of operation indicated that H2 and propionate-utilising SRB had outcompeted hydrogenophilic methanogens and propionate syntrophs. Acetoclastic methanogens were shown to play the dominant role in acetate conversion. Butyrate and ethanol-degrading syntrophs also remained active in the digester after 5 years of operation. Laboratory-scale hybrid reactor treatment at 55 degrees C of a diluted molasses influent, with and without sulphate supplementation, showed that the reactors could be operated with high stability at volumetric loading rates of 24 kgCOD.m-3.d-1 (12 h HRT). In the presence of sulphate (2 g/l-1; COD/sulphate ratio of 6:1), acetate conversion was severely inhibited, resulting in effluent acetate concentrations of up to 4000 mg.l-1.
This study investigated the hypothesis that antiformin-citric acid chemical curettage combined with scaling and root planing (SRP/AF) would significantly reduce pathogenic bacteria of the periodontal pocket when compared to scaling and root planing alone (SRP). Ten patients with moderate periodontitis participated. Four pockets per patient, one in each posterior sextant, were chosen. Pocket depths, attachment levels, and gingival index (GI) were measured. Bacterial samples were examined under dark-field microscope for: (1) spirochetes, (2) motile rods, and (3) all others. A randomized four-sextant treatment design was used. One sextant received no treatment, a second received SRP, and the two remaining sextants received identical treatments of SRP/AF. Of these two sextants, one was sampled at baseline and 12 weeks only in order to serve as an additional control. The remaining sextants were sampled at 1, 4, 8, and 12 weeks. Both SRP and SRP/AF significantly reduced the percentage of spirochetes and motile rods. A significant reduction in GI and pocket depths was also noted. Attachment level showed no significant change. Results of both treatment groups showed no significant differences among any of the parameters. A comparison of the two SRP/AF groups indicated no significant difference among clinical or bacterial parameters due to sampling.
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Previous research in our laboratory has shown that citric acid (CA) improves phytate P utilization in New Hampshire x Columbian (NHC) crossbred chicks fed a P-deficient corn-soybean meal diet. The current study was conducted to determine if CA is also effective in commercial broiler chicks (Ross x Ross). In 3 experiments, 4 replicate groups of 5 male NHC chicks and male commercial chicks were fed corn-soybean meal diets varying in CA and nonphytate P (NPP) from 8 to 22 d of age. In experiment 1, a 2 x 2 x 2 factorial treatment arrangement was used to evaluate the effect of 2 levels of CA (0 and 3%) and NPP (0.13 and 0.28%) in NHC chicks and commercial chicks. The commercial chicks, but not the NHC chicks, fed the 0.13% NPP diet had to be removed from the experiment after 3 to 5 d due to very poor growth and severe leg problems. Chick weight gain and tibia ash were significantly increased (P < 0.05) by CA in both types of chicks. In experiment 2, the same 2 x 2 x 2 factorial treatment arrangement was again used except that the NPP levels were 0.18 and 0.28%. Tibia ash was increased significantly (P < 0.05) with the addition of CA in both breeds of chicks; response was greater at 0.18% NPP than at 0.28% NPP. In experiment 3, graded levels of CA (0, 1, 2, 3, and 4%) were evaluated in commercial chicks fed diets containing 0.18% NPP. Tibia ash increased linearly (P < 0.05) as CA increased from 0 to 4%. The average increase in bone ash resulting from 3% CA supplementation in experiments 2 and 3 was 41%. These results indicate that CA markedly improved phytate P utilization in NHC and Ross x Ross commercial broiler chicks.