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Biosynthesis from gluconate of a random copolyester consisting of 3-hydroxybutyrate and medium-chain-length 3-hydroxyalkanoates by Pseudomonas sp. 61-3.

Pseudomonas sp. 61-3 isolated from soil was found to produce a polyester consisting of 3-hydroxyalkanoic acids of even carbon numbers C4, C6, C8, C10 and C12 when sodium gluconate was fed as the sole carbon source. The polyester produced was fractionated with boiling acetone. The acetone-insoluble fraction (28 wt%) of the polyester was a poly(3-hydroxybutyrate) homopolymer, while the acetone-soluble fraction (72 wt%) was composed of seven different 3-hydroxyalkanoate (3HA) units ranging from C4 to C12: 40 mol% 3-hydroxybutyrate (3HB), 5 mol% 3-hydroxyhexanoate (3HH), 20 mol% 3-hydroxyoctanoate (3HO), 24 mol% 3-hydroxydecanoate (3HD), 1 mol% 3-hydroxy-5-cis-decenoate (3H5D), 4 mol% 3-hydroxydodecanoate (3HDD) and 6 mol% 3-hydroxy-5-cis- dodecenoate (3H5DD). The copolymer was characterized by nuclear magnetic resonance (NMR) spectroscopy, gel permeation chromatography and differential scanning calorimetry. The acetone-soluble fraction of this amorphous copolymer was shown to have a random sequence distribution of the seven 3HA units of C4 to C12 by analysis of the 150 MHz 13C-NMR spectrum. This is the first example of microbial synthesis of a random copolyester consisting of 3HB and medium-chain-length 3HA units.

3-Hydroxybutyric Acid↗

Shortening and elongation of isolated outer hair cells in response to application of potassium gluconate, acetylcholine and cationized ferritin.

Individual outer hair cells isolated from guinea pig cochleae were observed in vitro during the application of solutions that are known to cause hair cells to shorten. Solutions containing high potassium, which depolarizes cells, were applied in the form of potassium gluconate. The initial response was a shortening, followed by an elongation, after which the hair cells nearly resumed their original length. Solutions containing the presumed efferent neurotransmitter acetylcholine also caused an initial shortening, occasionally followed by an elongation, where a cell either returned to normal or exceeded its original length. Solutions containing cationized ferritin caused some cells to shorten and caused others to lengthen. The results indicate that the hair cell response to a chemical stimulus can be bidirectional. Moreover, the initial response of an individual cell may depend not only on the stimulus but also on the physiological state of the hair cell or the original location of the hair cell along the length of the sensory epithelium when it was in the cochlea.

Acetylcholine↗

Multinuclear NMR study of the complexes of 6-phospho-D-gluconic acid with W(VI) and Mo(VI).

Multinuclear ((1)H, (13)C, (17)O, (31)P, (95)Mo, (183)W) magnetic resonance spectroscopy (1D and 2D) has been used to show that 6-phospho-d-gluconic acid forms three complexes with tungsten(VI) and six complexes with molybdenum(VI) in aqueous solution, depending on pH and concentration. Two isomeric 1:2 (metal-ligand) complexes are detected both with tungstate(VI) and molybdate(VI), having MO(2)(2+) centres and involving the carboxylate and the adjacent OH groups in addition to one 2:1 (metal-ligand) complex possessing a M(2)O(5)(2+) centre, with the ligand being coordinated by the carboxylate group and the three consecutive OH groups in positions 2, 3 and 4. Molybdate(VI) forms three additional species, which are not detected with tungstate. One of them is a 2:1 complex with a Mo(2)O(5)(2+) centre, with the ligand being tetradentate via O-3, O-4, O-5 and the phosphate group. The other two are 12:4 species, which can be seen as two 1:2 complexes bound together in a ring through two diphosphomolybdate moieties each derived from heptamolybdate by inclusion of two phosphate groups from the ligands.

Carbohydrate Conformation↗

Effectiveness of chlorhexidine gluconate-impregnated dressings in preventing central line-associated bloodstream infection in a paediatric intensive care unit: A randomised controlled trial.

OBJECTIVES: To compare the effectiveness and dermatological safety of chlorhexidine gluconate (CHG)-impregnated dressings versus conventional transparent dressings in preventing central line-associated bloodstream infection (CLABSI) in paediatric patients admitted to a PICU. METHODS: A single-blind, randomised controlled trial with a 1:1 allocation ratio was conducted in a tertiary PICU. A total of 250 paediatric patients with a central venous catheter were included (125 per group). Patient characteristics, catheter-related variables, catheter dwell time and skin alterations were recorded. The primary outcome was CLABSI, defined according to CDC/NHSN criteria. Bivariate analyses and multivariable logistic regression were performed, including clinically relevant covariates related to catheter exposure, dressing type, and vaccination status. The trial was registered at ClinicalTrials.gov (NCT07175116). RESULTS: CLABSI incidence was significantly lower in the CHG group compared with the conventional dressing group (2.4% vs. 18.4%; p&#xa0;<&#xa0;0.001). In multivariable analysis, CHG-impregnated dressings were independently associated with reduced CLABSI risk (OR = 0.15; 95% CI: 0.04-0.58). Longer catheter dwell time (OR = 1.13 per day; 95% CI: 1.04-1.22) and absence of complete vaccination (OR = 8.68; 95% CI: 2.48-30.47) were also associated with increased infection risk. Incomplete vaccination showed a similar trend without reaching statistical significance (OR = 6.48; 95% CI: 0.95-44.20). Skin alterations were more frequent in the CHG group (16.8% vs. 11.2%), predominantly in younger infants. CONCLUSIONS: CHG-impregnated dressings were associated with a significant reduction in CLABSI incidence in critically ill paediatric patients, although their use was linked to a modest increase in mild skin reactions. IMPLICATIONS FOR CLINICAL PRACTICE: CHG-impregnated dressings may be considered as part of a multifaceted infection prevention strategy, alongside appropriate catheter management, minimisation of catheter manipulation and proactive skin monitoring, with cautious and individualised use in infants.

Humans↗

Skewing of cytokine profiles towards T helper cell type 2 response in visceral leishmaniasis patients unresponsive to sodium antimony gluconate.

Thirty-two parasitologically confirmed visceral leishmanisis (VL) patients and 23 healthy, age- and gender-matched controls were recruited between April 2001 and March 2002 and studied for intracellular cytokine production after stimulation in vitro using a Fluorescence Activated Cell Sorter-based intracellular cytokine assay. The VL patients were given i.m. sodium antimony gluconate at a dose of 20 mg/kg bodyweight daily for 28 d and were grouped as responders (n = 11) or non-responders (no response after 28 d of treatment; n = 21). Clinically, the non-responders had longer duration of illness (P < 0.05), larger spleen size (P < 0.05), and higher parasite load (P < 0.05) than responders. The percentage of T helper (Th) cells producing interferon-gamma (IFN-gamma) was significantly higher (P < 0.001) in responders than non-responders. Non-responders had higher IFN-gamma production than control subjects (P < 0.001). The percentage of Th cells producing interleukin-4 (IL-4) was significantly higher in non-responders than responders (P = 0.003) as well as in healthy subjects (P < 0.001). The frequency of IL-4 producing cells in responders and control subjects was similar (P= 0.65). The cytokine polarization index, as calculated by the formula loge IFN-gamma producing cells/loge IL-4 producing cells, was significantly lower in non-responders compared with both responders and control subjects (P = 0.003 and P < 0.001, respectively). The overall cytokine bias in non-responders was skewed towards a IL-4 dominance or Th2-like response and this was primarily due to induction of IL-4.

Adolescent↗

The role of calcium gluconate in the treatment of hydrofluoric acid eye burn.

Hydrofluoric acid is used for different purposes in industry and in the home as a rust remover. Most exposures are accidental and may result in severe superficial and deep tissue injury as well as systemic toxicity. There is uncertainty regarding the optimal treatment of hydrofluoric acid eye injury. A patient in whom a solution of 49% hydrofluoric acid induced a large corneal erosion is described. Repeated instillation of 1% calcium gluconate eye drops combined with the conventional treatment of acid eye burns resulted in a complete and quick recovery.

Adult↗

Optimizing an artificial metabolic pathway: engineering the cofactor specificity of Corynebacterium 2,5-diketo-D-gluconic acid reductase for use in vitamin C biosynthesis.

The strict cofactor specificity of many enzymes can potentially become a liability when these enzymes are to be employed in an artificial metabolic pathway. The preference for NADPH over NADH exhibited by the Corynebacterium 2,5-diketo-D-gluconic acid (2,5-DKG) reductase may not be ideal for use in industrial scale vitamin C biosynthesis. We have previously reported making a number of site-directed mutations at five sites located in the cofactor-binding pocket that interact with the 2'-phosphate group of NADPH. These mutations conferred greater activity with NADH upon the Corynebacterium 2,5-DKG reductase [Banta, S., Swanson, B. A., Wu, S., Jarnagin, A., and Anderson, S. (2002) Protein Eng. 15, 131-140; (1)]. The best of these mutations have now been combined to see if further improvements can be obtained. In addition, several chimeric mutants have been produced that contain the same residues as are found in other members of the aldo-keto reductase superfamily that are naturally able to use NADH as a cofactor. The most active mutants obtained in this work were also combined with a previously reported substrate-binding pocket double mutant, F22Y/A272G. Mutant activity was assayed using activity-stained native polyacrylamide gels. Superior mutants were purified and subjected to a simplified kinetic analysis. The simplified kinetic analysis was extended for the most active mutants in order to obtain the kinetic parameters in the full-ordered bi bi rate equation in the absence of products, with both NADH and NADPH as cofactors. The best mutant 2,5-DKG reductase produced in this work was the F22Y/K232G/R238H/A272G quadruple mutant, which exhibits activity with NADH that is more than 2 orders of magnitude higher than that of the wild-type enzyme, and it retains a high level of activity with NADPH. This new 2,5-DKG reductase may be a valuable new catalyst for use in vitamin C biosynthesis.

Alanine↗

Effect of gluconic acid consumption during simulation of biological aging of sherry wines by a flor yeast strain on the final volatile compounds.

Flor yeast Saccharomyces cerevisiae (capensis G1) strain assimilates gluconic acid during the aerobic biological aging process of sherry wines and exerts significant changes on the final volatile compounds of wines, especially a decrease in volatile acidity and butanoic, isobutanoic, 2-methylbutanoic, and 3-methylbutanoic acids. This decrease may have a favorable effect on the quality of sherry wines.

Butyrates↗

Changes in gluconic acid, polyols and major volatile compounds in sherry wine during aging with submerged flor yeast cultures.

The traditional biological process by which sherry wines are aged can be accelerated by using submerged Saccharomyces cerevisiae var. capensis (G1) strain cultures previously grown in glycerol. The used controlled shaking conditions raise the acetaldehyde, acetoin, and meso 2,3-butanediol contents in the wine, and increases the consumption of gluconic acid by flor yeast relative to traditional biological aging under flor yeast velum.

Cell Culture Techniques↗

Changes in volatile compounds and aromatic series in sherry wine with high gluconic acid levels subjected to aging by submerged flor yeast cultures.

Volatile compounds of sherry wine containing gluconic acid under aging by submerged flor yeast cultures were analyzed. The aroma profile was obtained by grouping the compounds in nine aromatic series. The balsamic, fatty, herbaceous and empyreumatic series increased significantly as consequence of the increase of pantolactone, acids (butanoic, 2-methylbutanoic and 3-methylbutanoic), methionol and gamma-butyrolactone compounds, respectively. The decrease of higher alcohols promoted solvent series diminished. These changes are consistent with those observed in the production of commercial sherry wine using traditional biological aging.

Fermentation↗

Lithium gluconate 8% vs ketoconazole 2% in the treatment of seborrhoeic dermatitis: a multicentre, randomized study.

BACKGROUND: Lithium significantly improved seborrhoeic dermatitis symptoms in comparison with placebo. Objectives This randomized controlled trial was designed to show a non-inferiority of 15% (primary end-point) of lithium gluconate 8% ointment compared with ketoconazole 2% emulsion. METHODS: The study population comprised out-patients who had facial seborrhoeic dermatitis for at least 2 months, with moderate to severe erythema and desquamation at inclusion. The primary end-point was complete remission, defined as the disappearance of both erythema and desquamation. The non-inferiority of lithium was assessed on the 95% confidence interval (CI) of the difference between treatments. RESULTS: The intent-to-treat analysis (ITT) involved 288 patients and the per protocol (PP) analysis 269 patients. Treatment groups were comparable at baseline on age, sex, disease duration and symptoms. For the main criterion, the success rate was 52.0% (lithium) vs. 30.1% (ketoconazole) in the ITT population and 53.2% (lithium) vs. 30.7% (ketoconazole) in the PP population. The non-inferiority of lithium was demonstrated with differences of 21.9% (95% CI 10.0-33.7%) and 22.5% (95% CI 10.2-34.8%), respectively, in the ITT and PP population. As the lower limit of the 95% CI was > 0, the superiority of lithium was shown. Lithium also showed better results on other symptoms: burning and dryness. Adverse events were reported by 26.3% (lithium) and 25% (ketoconazole) of patients. CONCLUSIONS: Lithium was 22% more effective than ketoconazole in giving complete remission of seborrhoeic dermatitis, with comparable safety.

Adolescent↗

Amphotericin B is superior to sodium antimony gluconate in the treatment of Indian post-kala-azar dermal leishmaniasis.

Parasitologically confirmed cases of post-kala-azar dermal leishmaniasis (PKDL) were treated by infusion with amphotericin B deoxycholate (ABD; 1 mg/kg.day on days 1-20, 21-40 and 61-80) or by intramuscular injection with sodium antimony gluconate (SAG; 20-day courses at 20 mg/kg day, with 20-day, drug-free intervals). Of the 11 patients given ABD, all were cured with the three courses, none relapsed in 12 months of follow-up, all developed mild adverse effects (shivering and fever) because of the infusion, five lost their appetites, and three showed increases in their serum creatinine concentrations (although none exceeded 'normal' limits). In contrast, only seven (63%) of the 11 patients given SAG were considered treatment successes (improvement in lesions by the end of the third course) and these took six courses (two cases), nine courses (four cases) or 10 courses (one case) to cure completely. Two of the patients given SAG developed arthralgia and two others developed non-specific ST changes in their electrocardiograms (ECG), although their ECG were normal between courses. The better cure rate with ABD was not statistically significant, probably because of the small sample size. However, ABD appears to be a superior to SAG in terms of the speed of response and cure, although it is more expensive and has some nephrotoxicity. As the effectiveness of SAG against PKDL is apparently declining over time and the cost of ABD is prohibitive in poor countries such as India, a safe, cheap and more effective drug for the treatment of PKDL is needed.

Adolescent↗

The effect of prostaglandin E2 and calcium gluconate on orthodontic tooth movement and root resorption in rats.

Possible modifications in orthodontic tooth movement (OTM) and root resorption as a result of local injections of prostaglandin E2 (PGE2) alone and with calcium gluconate (Ca) formed the aim of the present study. Twenty-four 8-week-old male Wistar rats were selected and randomly divided into three groups of eight. Both quadrants of the upper jaws of the first group of animals were used; therefore this group comprised two groups: control and normal. The upper left first molars of these eight animals were not placed under orthodontic force and received no injection, to serve as the normal group, considered for root resorption comparison only. The control group had localized submucosal injections of normal saline on the buccal side of the upper right first molar. In the third group, 0.1 ml of 1 mg/ml PGE2 was injected at the same site and the fourth group received an intraperitoneal injection of 200 mg/kg Ca (10%) in addition to the PGE2. All the injections were performed on days 0 and 7. The orthodontic appliance consisted of a closed coil spring ligated to the upper right first molar and incisor, exerting a force of 60 g during the 21-day experimental period, after which the animals were sacrificed. Palatal halves were removed for histological examination and for calculation of the amount of root resorption. Statistical analysis of data showed a significant (P < 0.05) acceleration in OTM after PGE2 injection compared with the control group. The addition of Ca reduced OTM but a significant increase (P< 0.05) was still recorded. A significant difference (P < 0.05) in root resorption was only observed between the PGE2 and normal groups. The findings show the importance of calcium ions working in association with PGE2 in stabilizing root resorption while significantly increasing OTM.

Analysis of Variance↗

Dietary sodium gluconate protects rats from large bowel cancer by stimulating butyrate production.

Butyrate has an antitumorigenic effect on colorectal cancer cell lines. Dietary sodium gluconate (GNA) promotes butyrate production in the large intestine. Accordingly, we examined the effect of dietary GNA on tumorigenesis in the large intestine in rats. Male Fisher-344 rats (n = 32) were divided into 4 groups: 2 diets (with or without 50 g GNA/kg basal diet) x 2 treatments (with or without carcinogen administration). Colonic tumors were induced by 3 intraperitoneal injections of azoxymethane (15 mg/kg body wt, 1 time/wk) and dietary deoxycholic acid (2 g/kg basal diet). The experiment was conducted for 33 wk except for a few rats. Ingestion of GNA increased cecal butyrate concentration at the end of experiment (P < 0.01). No tumor development occurred in the untreated groups. Ingestion of GNA decreased the incidence of tumors in rats administered the carcinogen (37.5 vs. 100%, P < 0.05). Ingestion of GNA also decreased the mean number of tumors per rat (0.5 +/- 0.8 vs. 2.8 +/- 1.5, P < 0.01). beta-Catenin accumulation and TdT-mediated dUTP nick end labeling (TUNEL) positive cells in tumors were histochemically examined. The results of this study suggested that the antitumorigenic effect of GNA may involve the stimulation of apoptosis through enhanced butyrate production in the large intestine.

Adenocarcinoma↗

Identification and nucleotide sequence of the promoter region of the Bacillus subtilis gluconate operon.

The nucleotide sequence (742 bp) of the promoter region of the Bacillus subtilis gluconate (gnt) operon is presented. Nuclease Sl mapping revealed the start point of the transcription and suggested that the expression of this operon is probably regulated at the transcriptional level. The sequences of the -35 and -10 regions suggested that RNA polymerase possessing sigma-43 may recognize this structure. The 223 bp fragment containing 100 bp upstream from the transcription start site actually exhibited a promoter activity when cloned in a promoter probe vector of pPL603B. This promoter activity was highly derepressed and although still under catabolite repression. The fragment on a high copy plasmid could titrate a regulator of the gnt operon so that the expression of the operon on the host chromosome also became derepressed.

Amino Acid Sequence↗

Alteration of the specificity of the cofactor-binding pocket of Corynebacterium 2,5-diketo-D-gluconic acid reductase A.

The NADPH-dependent 2,5-diketo-D-gluconic acid (2,5-DKG) reductase enzyme is a required component in some novel biosynthetic vitamin C production processes. This enzyme catalyzes the conversion of 2,5-DKG to 2-keto-L-gulonic acid, which is an immediate precursor to L-ascorbic acid. Forty unique site-directed mutations were made at five residues in the cofactor-binding pocket of 2,5-DKG reductase A in an attempt to improve its ability to use NADH as a cofactor. NADH is more stable, less expensive and more prevalent in the cell than is NADPH. To the best of our knowledge, this is the first focused attempt to alter the cofactor specificity of a member of the aldo-keto reductase superfamily by engineering improved activity with NADH into the enzyme. Activity of the mutants with NADH or NADPH was assayed using activity-stained native polyacrylamide gels. Eight of the mutants at three different sites were identified as having improved activity with NADH. These mutants were purified and subjected to a kinetic characterization with NADH as a cofactor. The best mutant obtained, R238H, produced an almost 7-fold improvement in catalysis with NADH compared with the wild-type enzyme. Surprisingly, most of this catalytic improvement appeared to be due to an improvement in the apparent kcat for the reaction rather than a large improvement in the affinity of the enzyme for NADH.

Binding Sites↗

Sugar-free zinc gluconate glycine lozenges (Cold-Eeze) do not adversely affect glucose control in patients with type 1 or type 2 diabetes mellitus.

Several controlled clinical trials have shown that zinc gluconate glycine lozenges can reduce symptom severity and duration of symptoms in patients with the common cold. Over-the-counter zinc lozenges are used commonly by the general population, including people with diabetes. The purpose of this study was to assess the effects of sugar-free Cold-Eeze (The Quigley Corp., Doylestown, PA), a commonly used zinc preparation, on glucose control in patients maintained on stable antidiabetic therapy. Forty-eight patients with either type 1 (n = 3) or type 2 (n = 45) diabetes were randomized in a 3:1 ratio to receive either zinc lozenges (four to six lozenges/day for 10 days) or matching placebo. The primary endpoint was change in serum fructosamine concentration. Secondary endpoints included daily home glucose and fasting blood glucose monitoring (baseline, days 10 and 21). The mean age for all patients was 54 years (range, 25 to 76), with slightly more women (60%). The treatment groups did not differ with respect to age, sex, body mass index, duration of diabetes, baseline hemoglobin A1C level, or fasting plasma glucose level. The patients treated with placebo (n = 13) and zinc (n = 34) had similar fructosamine levels (mean +/- SD) at baseline (318 +/- 90 versus 297 +/- 86 micromol/l, respectively). After 10 days of dosing, both groups showed modest reductions in serum fructosamine (-7 +/- 42 and -9 +/- 90 micromol/l). These changes were not statistically significant. In conclusion, these findings suggest that sugar-free zinc lozenges can be administered safely to patients with diabetes without deleterious effects on glycemic control.

Administration, Oral↗