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Impact of vesicoureteral reflux on the size of renal lesions after an episode of acute pyelonephritis.

PURPOSE: We determined the impact of vesicoureteral reflux (VUR) on the size of renal lesions in children after an episode of acute pyelonephritis. MATERIALS AND METHODS: A total of 161 children (mean age 2.44 years) with acute pyelonephritis were studied. All had renal lesions on dimercapto-succinic acid scintigraphy done at admission to the hospital. A second dimercapto-succinic acid scan was performed at 3 months. Voiding cystourethrography was done at 6 weeks and VUR was graded I to V. For each renal unit layouts of renal lesions were drawn, and the damage surface was calculated and reported for the total surface of the kidney. RESULTS: Mean size of acute lesions and scars increased with severity of reflux (p <0.0001), with an important overlap of individual values. Mean size of renal scars in the group of renal units with acute lesions was 5.8% +/- 8.5% in patients without VUR, 9.9% +/- 7.3% in those with grade I reflux, 7.7% +/- 11.0% in those with grade II reflux, 17.7% +/- 14.7% in those with grade III reflux and 17.4% +/- 27.7% in those with grade IV reflux (p <0.001). The size of renal lesions decreased significantly with time. The rate of regression of lesions decreased with increasing reflux. When analyzed according to 3 age groups sizes of scars increased significantly with age. CONCLUSIONS: VUR has an impact on the size of renal lesions after an episode of pyelonephritis. Children with a grade III or IV reflux are more likely to have larger renal scars. On the other hand, acute lesions of important size may develop even in the absence of VUR.

Acute Disease↗

pH-dependent and carrier-mediated transport of salicylic acid across Caco-2 cells.

The transport of monocarboxylic acid drugs such as salicylic acid was examined in the human colon adenocarcinoma cell line, Caco-2 cells that possess intestinal epithelia-like properties. [14C]Salicylic acid transport was pH-dependent and appeared to follow the pH-partition hypothesis. However, 10 mM unlabelled salicylic acid significantly reduced the permeability coefficient of [14C]salicylic acid. Kinetic analysis of the concentration dependence of the permeation rate of salicylic acid across Caco-2 cells showed both saturable (Kt = 5.28 +/- 0.72 mM Jmax = 36.6 +/- 3.54 nmol min-1 (mg protein)-1) and nonsaturable (kd = 0.37 +/- 0.08 microL min-1 (mg protein)-1) processes. The permeation rate of [14C]salicylic acid was competitively inhibited by both acetic acid and benzoic acid, which were demonstrated in our previous studies to be transported in the carrier-mediated-transport mechanism which is responsible for monocarboxylic acids. Furthermore, certain monocarboxylic acids significantly inhibited [14C]salicylic acid transport, whereas salicylamide and dicarboxylic acids such as succinic acid did not. From these results, it was concluded that the transcellular transport of [14C]salicylic acid across Caco-2 cells is by the pH-dependent and carrier-mediated transport mechanism specific for monocarboxylic acids.

Acetates↗

Utilization of Lactic Acid by Fusarium oxysporum var. lini: Regulation of Transport and Metabolism.

Lactic acid was transported in Fusarium oxysporum var. lini ATCC 10960 by a saturable transport system that had a half-saturation constant of 56.6 +/- 7.5 muM and a maximum velocity of 0.61 +/- 0.10 mmol h g (dry weight) at 26 degrees C and pH 5.0. This transport system was inducible and was not expressed in the presence of a repressing substrate. Evidence is presented that the anionic form lactate was taken up by the cells. Propionic, acetic, pyruvic, and bromoacetic acids but not succinic acid competitively inhibited the transport of lactic acid. Bromoacetic acid, which was not metabolized, was taken up to a steady-state level when intracellular and extracellular concentrations were identical, indicating that the transport system was not accumulative. The enzymatic activity that was physiologically more relevant in the metabolism of lactic acid was lactate: ferricytochrome c oxidase. This enzyme did not exhibit stereospecifity and was induced by lactic acid.

Journal Article↗

Isolation and characterization of a novel toluene-degrading, sulfate-reducing bacterium.

A novel sulfate-reducing bacterium isolated from fuel-contaminated subsurface soil, strain PRTOL1, mineralizes toluene as the sole electron donor and carbon source under strictly anaerobic conditions. The mineralization of 80% of toluene carbon to CO2 was demonstrated in experiments with [ring-U-14C]toluene; 15% of toluene carbon was converted to biomass and nonvolatile metabolic by-products, primarily the former. The observed stoichiometric ratio of moles of sulfate consumed per mole of toluene consumed was consistent with the theoretical ratio for mineralization of toluene coupled with the reduction of sulfate to hydrogen sulfide. Strain PRTOL1 also transforms o- and p-xylene to metabolic products when grown with toluene. However, xylene transformation by PRTOL1 is slow relative to toluene degradation and cannot be sustained over time. Stable isotope-labeled substrates were used in conjunction with gas chromatography-mass spectrometry to investigate the by-products of toluene and xylene metabolism. The predominant by-products from toluene, o-xylene, and p-xylene were benzylsuccinic acid, (2-methylbenzyl)succinic acid, and 4-methylbenzoic acid (or p-toluic acid), respectively. Metabolic by-products accounted for nearly all of the o-xylene consumed. Enzyme assays indicated that acetyl coenzyme A oxidation proceeded via the carbon monoxide dehydrogenase pathway. Compared with the only other reported toluene-degrading, sulfate-reducing bacterium, strain PRTOL1 is distinct in that it has a novel 16S rRNA gene sequence and was derived from a freshwater rather than marine environment.

Acetyl Coenzyme A↗

Effect of uremic toxins--guanidino compounds and creatinine--on proliferation of HL60 and K562 cell lines.

Growth of the promyelocytic cell line HL60 and the erythroleukemia cell line K562 is inhibited by 'uremic toxins': creatinine, guanidino propionic acid and guanidino succinic acid in a concentration range similar to that of uremic sera. Among the tested compounds, creatinine exhibits the strongest and most dose-dependent inhibitory effect on both kinds of cells. These results provide a better understanding of the mechanism involved in the anemia of uremic patients.

Cell Line↗

Biofilm reactors for industrial bioconversion processes: employing potential of enhanced reaction rates.

This article describes the use of biofilm reactors for the production of various chemicals by fermentation and wastewater treatment. Biofilm formation is a natural process where microbial cells attach to the support (adsorbent) or form flocs/aggregates (also called granules) without use of chemicals and form thick layers of cells known as "biofilms." As a result of biofilm formation, cell densities in the reactor increase and cell concentrations as high as 74 gL(-1) can be achieved. The reactor configurations can be as simple as a batch reactor, continuous stirred tank reactor (CSTR), packed bed reactor (PBR), fluidized bed reactor (FBR), airlift reactor (ALR), upflow anaerobic sludge blanket (UASB) reactor, or any other suitable configuration. In UASB granular biofilm particles are used. This article demonstrates that reactor productivities in these reactors have been superior to any other reactor types. This article describes production of ethanol, butanol, lactic acid, acetic acid/vinegar, succinic acid, and fumaric acid in addition to wastewater treatment in the biofilm reactors. As the title suggests, biofilm reactors have high potential to be employed in biotechnology/bioconversion industry for viable economic reasons. In this article, various reactor types have been compared for the above bioconversion processes.

Journal Article↗

Inhibitory effects of RRR-alpha-tocopheryl succinate on benzo(a)pyrene (B(a)P)-induced forestomach carcinogenesis in female mice.

AIM: To study the inhibitory effects of VES (RRR-alpha-tocopheryl Succinate, VES),a derivative of natural Vitamin E, on benzo(a)pyrene(B(a)P)-induced forestomach tumor in female mice. METHODS: The model of B(a)P-induced forestomach tumor was established according to the methods of Wattenberg with slight modify-cations. One hundred and eighty female mice (6 weeks old) were divided into six groups equally; negative control (Succinic acid), vehicle control (Succinate+B(a)P),positive control(B(a)P), high VES(2.5 g/kg.b.w+B(a)P), low VES(1.25 g/kg.b.w+B(a)P)ig as well as VES by ip (20 mg/kg.b.w+B(a)P). Except the negative control group, the mice were administrated with B(a)P ig. and corresponding treatments for 4 weeks to study the anti-carcinogenetic effect of VES during the initiation period. The experiment lasted 29 weeks, in which the inhibitory effects of VES both on tumor incidence and tumor size were tested. RESULTS: The models of B(a)P-induced forestomach tumor in female mice were established successfully. Some were cauliflower-like, others looked like papilla, even a few were formed into the ulcer cavities. VES at 1.25 g/kg.b.w, 2.5 g/kg.b.w. by ig and 20 mg/kg.b.w. via ip could decrease the number of tumors per mouse (1.7 plus minus 0.41, 1.6 plus minus 0.34 and 1.1 +/- 0.43), being lower than that of B(a)P group (5.4 +/- 0.32, P<0.05). The tumor incidence was inhibited by 18.2%, 23.1% and 50.0%. VES at 1.25 g/kg.b.w., 2.5 g/kg.b.w. by ig and 20 mg/kg.b.w. via ip reduced the total volume of tumors per mouse (54.8 +/- 8.84, 28.4 8 +/- 8.32 and 23.9 8 +/- 16.05), being significantly lower than that of B(a)P group (150.2 8 +/- 20.93, P < 0.01). The inhibitory rates were 63.5%, 81.1% and 84.1%, respectively. CONCLUSION: VES has inhibitory effects on B(a)P-induced forestomach carcinogenesis in female mice, especially by ip and it may be a potential anti-cancer agent in vivo.

Animals↗

[10-Acyldithranol dimers; new derivatives of the antipsoriatic dithranol; synthesis, characterization and biochemical properties].

With regard to the synthesis of 10-(omega-carboxyacyl)-derivatives dithranol 1 was reacted with the carboxylic acid dichlorides of succinic acid, glutaric acid, adipic acid and pimelic acid in toluene and collidin as a base. Instead of the expected derivatives the 10-acyldithranol dimers 5-7 were isolated as main products except for the reaction of succinyl chloride where only lactone 2 was formed. Moreover the 6-ring lactone 3 as a side product and traces of the 7-ring lactone 4 could also be isolated and characterized. All compounds revealed to be inhibitors of the enzyme glucose-6-phosphate dehydrogenase, indicating antipsoriatic activity.

Anthralin↗

[Role of GABA and its derivatives in regulating cerebral circulation].

GABA, its derivative -- gamma-hydroxybuturic acid and metabolite --succinic acid have a pronounced dilatatory activity on cerebral circulation in various brain parts. GABA increases cerebral circulation by 25.3%, gamma-hydroxybutyric acid by 35.9% and sucinic acid by 20.4%. In ischaemia of the brain a relationship has been established between cerebral circulation, changes in the GABA system in brain and in the walls of cerebral arteries. The content of GABA increases following enhancement of GAD activity and inhibition of GABA-T. The increase of endogenous GABA level in brain during hypoxia of the brain brings to an improvement of blood circulation through increasing collateral vessels. Experiments with GABA-T inhibition by aminooxyacetic acid give direct evidence about the role of the GABA system in cerebral blood circulation. This mechanism is evaluated by us as an example of an autoregulatory system that is realized by a feed-back mechanism providing the adaptability and compensatory function of cerebral haemodynamics to changing conditions.

4-Aminobutyrate Transaminase↗

Monitoring of itaconic acid hydrogenation in a trickle bed reactor using fiber-optic coupled near-infrared spectroscopy.

Near-infrared (NIR) spectroscopy has been applied to determine the conversion of itaconic acid in the effluent stream of a trickle bed reactor. Hydrogenation of itaconic to methyl succinic acid was carried out, with the trickle bed operating in recycle mode. For the first time, NIR spectra of itaconic and methyl succinic acids in aqueous solution, and aqueous mixtures withdrawn from the reactor over a range of reaction times, have been recorded using a fiberoptic sampling probe. The infrared spectra displayed a clear isolated absorption band at a wavenumber of 6186 cm(-1) (wavelength 1.617 microm) resulting from the =C-H bonds of itaconic acid, which was found to decrease in intensity with increasing reaction time. The feature could be more clearly observed from plots of the first derivatives of the spectra. A partial least-squares (PLS) model was developed from the spectra of 13 reference samples and was used successfully to calculate the concentration of the two acids in the reactor effluent solution. Itaconic acid conversions of 23-29% were calculated after 360 min of reaction time. The potential of FT-NIR with fiber-optic sampling for remote monitoring of three-phase catalytic reactors and validation of catalytic reactor models is highlighted in the paper.

Bioreactors↗

[Pathomorphological and histochemical changes in the liver of rabbits following fractional irradiation with Co-60].

Two groups of experimental animals, each consisting of 12 rabbits, were subjected to local fractional irradiation with cobalt 60. Group I received the total dose of 2580 R during 13 days, group II - 5100 R during 24 days. The effects of irradiation were estimated on the strength of histological examination of the liver immediately and after 7, 15, 30, 60 and 90 days after the last exposition. The histological sections were stained with haematoxylin and eosin, and colour reactions were performed for argentaffine and collagen fibres and for glycogen, neutral fats, alkaline and acid phosphatase, ATP-ase, glycose-6-phosphatase, non-specific esterase and succinic acid dehydrogenase. It was found that the dose of 2580 R is safe for the liver. The effects of irradiation were slight and limited to weak catabolic disturbances in the form of mild steatosis of the liver and of a transient and short-lived fall of glycogen and rise of hydrolytic enzymes. More pronounced and intense changes were observed in the other group of animals. During the early period, the changes were of a retrograde character and were typical of the acute post-irradiation effect. There was necrosis of the walls of the blood vessels, of the epithelium of the bile ducts and of the liver cells, accompanied by a rise in the hydrolytic enzymes and by a considerable fall of the level of glycogen and succinic acid dehydrogenase. During the late period (30-90 days), changes typical of the so-called post-irradiation hepatitis were found histologically.

Animals↗

Effect of carbon source on growth rate and phospholipid composition of Escherichia coli 15T- and an unsaturated fatty acid auxotroph.

Escherichia coli 15T- was grown with glucose, succinic acid, aspartic acid, oleic acid, and oleic plus aspartic acids as carbon sources, and a fatty acid auxotroph derived from 15T- was grown on oleic acid and oleic plus aspartic acids. The doubling time, phospholipid composition, phosphorus content, and the fatty acid composition of the phospholipids of cells in each of the media were determined. In all cases, phosphatidylethanolamine was the major phospholipid present; but with 15T- its concentration was inversely proportional to the doubling time in unsupplemented media. With the auxotroph the phosphatidylethanolamine concentration was essentially unchanged with growth. Total lipid phosphorus was inversely proportional to doubling time, an effect particularly evident with the auxotroph. Without oleic acid supplementation, the major effects of carbon source on fatty acid composition are decreases in the content of palmitoleic acid and increases in the content of cis-9, 10-methylene hexadecanoic acid as growth rate decreases. Oleic acid supplementation elevated 18:1 fatty acid content in both 15T- and the auxotroph.

Aspartic Acid↗

Succinate production in dual-phase Escherichia coli fermentations depends on the time of transition from aerobic to anaerobic conditions.

We examined succinic acid production in Escherichia coli AFP111 using dual-phase fermentations, which comprise an initial aerobic growth phase followed by an anaerobic production phase. AFP111 has mutations in the pfl, ldhA, and ptsG genes, and we additionally transformed this strain with the pyc gene (AFP111/pTrc99A-pyc) to provide metabolic flexibility at the pyruvate node. Aerobic fermentations with these two strains were completed to catalog physiological states during aerobic growth that might influence succinate generation in the anaerobic phase. Activities of six key enzymes were also determined for these aerobic fermentations. From these results, six transition times based on physiological states were selected for studying dual-phase fermentations. The final succinate yield and productivity depend greatly on the physiological state of the cells at the time of transition. Using the best transition time, fermentations achieved a final succinic acid concentration of 99.2 g/l with an overall yield of 110% and productivity of 1.3 g/l h.

Aerobiosis↗

The acid end-products of glucose metabolism of oral and other haemophili.

The acids produced in broth culture by various species of oral haemophili and by stock strains of capsulated and other haemophili were identified and measured by gas-liquid chromatography. Succinic acid was the major acid end-product of all strains, with acetic acid also being regularly produced but in smaller amounts. A stock strain, Haemophilus parainfluenzae NCTC 4101, produced less succinic acid than other strains of haemophili. Strain NCTC 4101 possessed all the enzymes of the tricarboxylic acid cycle, as previously reported, but in the other haemophili examined only succinic dehydrogenase, fumarase and malate dehydrogenase could be detected. No other enzymes of the tricarboxylic acid cycle were detected and isocitrate lyase, malate synthase and pyruvate carboxylase were also absent. Phosphoenolpyruvate-carboxylase was present in all strains. A partial tricarboxylic acid cycle and marked malate dehydrogenase activity appear to be characteristic of haemophili. The pathway to succinate in haemophili appears to be via carboxylation of phosphoenolpyruvate to oxalacetate and thence via malate and fumarate. The results of tracer studies on a single oral strain of H. parainfluenzae using various labelled substrates were in keeping with this proposed metabolic pathway.

Carboxylic Acids↗

Effects of medium composition on production of 5-aminolevulinic acid by recombinant Escherichia coli.

The recombinant Escherichia coli BL21(DE3) harboring hemA from Agrobacterium radiobacter, which was engineered in our previous work, was used for the extracellular production of 5-aminolevulinic acid (ALA). The effects of various physiological factors, such as the concentrations of precursors (glycine, succinic acid and glucose) and the inhibitor 5-aminolevulinate dehydratase (levulinic acid), on the ALA accumulation in the fermentation broth were investigated in both shake flasks and a jar fermentor. Among these precursors, glycine exhibited the strongest ability to inhibit cell growth, while glucose mainly inhibited ALA formation. The optimum initial concentrations of glycine, succinic acid and glucose were found to be 2.0, 10.0 and 2.0 g/l, respectively. Levulinic acid (LA; 30 mM) was fed to the fermentation broth at the end of the exponential cell growth phase (about 8 h), and the intracellular activity of ALA dehydratase was efficaciously suppressed. Repeating the optimum composition of the medium in a stirred tank fermenter resulted in 1.49 g/l ALA. Furthermore, the fed batch of the precursors and inhibitor further increased ALA production up to 3.01 g/l.

Aldehyde Oxidoreductases↗

Synergistic insulinotropic action of succinate, acetate, and glucose esters in islets from normal and diabetic rats.

Succinic acid esters are currently under investigation as possible insulinotropic tools in the treatment of noninsulin-dependent diabetes mellitus. The present article introduces three novel nutrient esters and aims mainly to explore, in both normal and GK rats, the secretory response to such esters when tested alone or in combination. It documents that in pancreatic islets from normal rats, methyl acetate (10 mM), which fails to augment basal insulin output, potentiates the secretory response to succinate dimethyl ester (also 10 mM). It also reveals that alpha-D-glucose pentaacetate (alpha GPA) (1.7 mM) stimulates insulin release in the absence of any other exogenous nutrient and even more so in the presence of succinate methyl ester. Moreover, the methyl esters of succinic acid (10 mM), when used together with either methyl acetate or alpha GPA, provoked insulin secretion in islets from diabetic GK rats incubated in the absence of D-glucose, although no significant secretory response of such islets could be detected when each of these agents was tested separately. These findings thus draw attention to the insulinotropic potential in type 2 diabetes of selected combinations of nutrient esters, including a D-glucose ester presumably able to enter into islet cells without requiring the intervention of a hexose carrier.

Acetates↗

Oxo-4-methylpentanoic acid directs the metabolism of GABA into the Krebs cycle in rat pancreatic islets.

OMP (oxo-4-methylpentanoic acid) stimulates by itself a biphasic secretion of insulin whereas L-leucine requires the presence of L-glutamine. L-Glutamine is predominantly converted into GABA (gamma-aminobutyric acid) in rat islets and L-leucine seems to promote its metabolism in the 'GABA shunt' [Fernández-Pascual, Mukala-Nsengu-Tshibangu, Martín del Río and Tamarit-Rodríguez (2004) Biochem. J. 379, 721-729]. In the present study, we have investigated how 10 mM OMP affects L-glutamine metabolism to uncover possible differences with L-leucine that might help to elucidate whether they share a common mechanism of stimulation of insulin secretion. In contrast with L-leucine, OMP alone stimulated a biphasic insulin secretion in rat perifused islets and decreased the islet content of GABA without modifying its extracellular release irrespective of the concentration of L-glutamine in the medium. GABA was transaminated to L-leucine whose intracellular concentration did not change because it was efficiently transported out of the islet cells. The L-[U-14C]-Glutamine (at 0.5 and 10.0 mM) conversion to 14CO2 was enhanced by 10 mM OMP within 30% and 70% respectively. Gabaculine (250 microM), a GABA transaminase inhibitor, suppressed OMP-induced oxygen consumption but not L-leucine- or glucose-stimulated respiration. It also suppressed the OMP-induced decrease in islet GABA content and the OMP-induced increase in insulin release. These results support the view that OMP promotes islet metabolism in the 'GABA shunt' generating 2-oxo-glutarate, in the branched-chain alpha-amino acid transaminase reaction, which would in turn trigger GABA deamination by GABA transaminase. OMP, but not L-leucine, suppressed islet semialdehyde succinic acid reductase activity and this might shift the metabolic flux of the 'GABA shunt' from gamma-hydroxybutyrate to succinic acid production.

Aldehyde Oxidoreductases↗