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Simultaneous production of catalase, glucose oxidase and gluconic acid by Aspergillus niger mutant.

The production of gluconic acid, extracellular glucose oxidase and catalase in submerged culture by a number of biochemical mutants has been evaluated. Optimization of stirrer speed, time cultivation and buffering action of some chemicals on glucose oxidase, catalase and gluconic acid production by the most active mutant, AM-11, grown in a 3-L glass bioreactor was investigated. Three hundred rpm appeared to be optimum to ensure good growth and best glucose oxidase production, but gluconic acid or catalase activity obtained maximal value at 500 or 900 rpm, respectively. Significant increase of dissolved oxygen concentration in culture (16-21%) and extracellular catalase activity were obtained when the traditional aeration was employed together with automatic dosed hydrogen peroxide.

Aspergillus niger↗

Effect of ammonium and nitrate ratio on glucose oxidase activity during gluconic acid fermentation by a mutant strain of Aspergillus niger.

Of the factors tested, the source and concentration of carbon and nitrogen in the medium exerted maximum effect on growth and acid production. Glucose (15%) and urea (0.14%) induced glucose oxidase synthesis and optimum yield of calcium gluconate. Potassium dihydrogen phosphate (0.2%) and magnesium sulphate (0.06%) stimulated glucose oxidase activity and calcium gluconate production. Borax at a concentration of 1.5 g/L induced maximum glucose oxidase and calcium gluconate production with increased glucose utilization.

Aspergillus niger↗

Calcinosis cutis following extravasation of calcium gluconate in neonates.

Neonatal hypocalcemia is not an uncommon condition, especially in the premature neonate. It is effectively treated by intravenous infusion with calcium gluconate. We treated nine neonates with subcutaneous calcium deposition following calcium replacement with calcium gluconate from Jan. 1997 to Dec. 1997. Three of the infants were born to diabetic mothers, two had perinatal asphyxia and four were born prematurity. The average dosing number was 7.4 (5 to 9 doses). The onset of calcinosis cutis was 5 to 11 days after the first dose. The replacement of calcium gluconate caused amorphous masses at the site of extravasation and contracture of joint movement. A radiographic study was performed to determine the extent and course of extravasation, and areas remote from the infusion site also showed calcification. There is no specific mode of treatment except supportive management and a skin graft. The patient could functionally recover with cosmetic residue. In our follow-up clinics, all infants completely recovered without functional limitations.

Calcinosis↗

Anosmia after intranasal zinc gluconate use.

BACKGROUND: Zinc is an essential mineral. Beneficial zinc absorption takes place via enteral, parenteral, or cutaneous routes. However, direct application to the olfactory epithelium has been reported to cause loss of smell. Recently, intranasal zinc gluconate has been recommended as a treatment for the common cold. Severe posttreatment hyposmia and anosmia have been observed. METHODS: The case report of a typical patient is presented and analyzed in detail, followed by a series of patients with severe hyposmia or anosmia after the use of intranasal zinc gluconate. RESULTS: Although interindividual variation in drug response and drug effect is apparent, the severe hyposmia or anosmia appears to be long lasting or permanent in some cases. The mechanism of olfactory loss is thought to be the direct action of the divalent zinc ion on the olfactory receptor cell. CONCLUSIONS: Zinc ions are toxic to olfactory epithelium. Reports of severe hyposmia with parosmia or anosmia have occurred after intranasal use of zinc gluconate.

Administration, Intranasal↗

[Gluconic acid forming enzymes in Aspergillus niger (author's transl)].

At least three gluconic acid forming enzymes were identified in cell-free extracts of Aspergillus niger: glucose oxidase (EC 1.1.3.4), a glucose dehydrogenase (EC 1.1.99.10), and an enzyme or a mixture of enzymes catalyzing the cleavage of 6-phosphogluconate into gluconate and inorganic phosphate. 2,6-dichlorphenolindophenol was one of the hydrogen acceptors in vitro of the glucose dehydrogenase. Some properties of this enzyme (Km values, pH-dependence, substrate and hydrogen acceptor specificity), as determined in cell-free extracts, were found to be in good agreement with properties described in literature for a glucose dehydrogenase which has been purified from Aspergillus oryzae. The formation of Pi from 6-phosphogluconate and other phosphate esters was found to have an optimum between pH 7 and 8 , and another below pH 4. This suggests that it is catalyzed by an alkaline and an acid phosphomonoesterase (EC 3.1.3.1, 3.1.3.2), both enzymes exhibiting only low substrate specificity. The influence of extraction and assay buffers on the activity of gluconate forming enzymes was investigated. Loss of activity during preparation of cell-free extracts, as calculated from loss of activity storage of cell-free extracts at 4 degrees C, was found to be lower than 4%. Purified glucose oxidase added before homogenization was found in the extract almost quantitatively.

Acid Phosphatase↗

Selection of lacZ operon fusions in genes of gluconate metabolism in E. coli. characterization of a gntT::lacZ fusion.

The initial steps involved in the utilization of gluconate by E. coli, its incorporation into the cell and subsequent phosphorylation to gluconate 6-phosphate, conform two systems that duplicate activities. These systems, GntI and GntII, are specified by two sets of genes distinctly regulated and located respectively at the malA-asd (75 min) and fdp-valS (96 min) regions of the bacterial chromosome. The presence of duplicate activities in the metabolism of gluconate of E. coli, has made difficult the study of the expression and participation of the GntI and GntII systems. In order to advance in this respect, the phage lambda placMu53 was used to select operon gnt::lacZ fusion in a E. coli strain delta (edd-zwf), delta (gnd-his), delta lac. Here we report the study of a gntT::lacZ fusion. This transductant allowed to differentiate the inducible expression of the gntT and gntU genes. Its characteristics, in agreement with previous reports, support the central role of the gntT gene in this metabolism.

Culture Media↗

Comparison trial of iron succinylprotein complex or iron gluconate complex in the treatment of iron deficiency anemia.

Forty patients suffering from iron deficiency anemia were randomly assigned to receive iron succinylprotein complex (containing 80 mg of elemental iron daily) or iron gluconate complex (containing 125 mg of elemental iron daily) for two months. Improvement, as measured by hematological variables, was noted in both groups, iron succinylprotein complex being superior to iron gluconate complex on some measures. Side effects were reported by two of the 20 patients treated with iron succinylprotein complex and by ten of the 20 patients treated with iron gluconate complex.

Adult↗

[Value of calcium gluconate labelled with 99mTc in the detection of intracranial tumors].

In a preliminary communication, the authors compare the success coefficients of pertechnate 99m-Tc and of a new labelled molecule: calcium gluconate 99m-Tc, in a group of 32 patients. The results obtained showed the undoubted superiority of the gluconate in comparison with the pertechnate in the detection of intracranial metestases, and in particular those of the posterior fossa. It proved equally effective as bleomycin labeled with 57-Co. Amongst the other types of tumour studies, the use of the gluconate seems to be particularly indicated for neoplasms of the posterior fossa and in those close to the base of the skull and venous sinuses, given the marked reduction in "background noise" obtained when this method is used.

Bleomycin↗

[Intravenous drop of calcium gluconate for phosphorus burns].

20 patients with phosphor burn (TBSA 2%-75%) were cured by i.v. drop of calcium gluconate combined with other therapies including eschar conservation. Our experimental data showed that dogs with burn by spreading 85% phosphoric acid and napalm locally increased the level of plasma phosphorus and pathological damages to the heart, lung, kidney and etc were similar to those previously reported phosphorus burns. Intravenous drop of calcium gluconate after phosphate burn reduced the level of plasma phosphorus to normal rapidly and lessened the visceral damages. We consider that i.v. drop of calcium gluconate can accelerate the elimination of phosphorus, and prevent phosphorus poisoning after phosphorus burns.

Adolescent↗

Protective effect of zinc gluconate on chemically induced gastric ulcer.

This study was undertaken in rats to ascertain the role of zinc as an antiulcerogenic agent employing its more bioavailable gluconate derivative. Pretreatment with zinc gluconate 10 mg/kg body wt orally for three consecutive days, protected against alcohol induced gastric epithelial damage and also significantly prevented non-steroidal anti-inflammatory drugs (NSAID) induced gastric ulcer in rats. The enhanced levels of mucus, and hexosamine and decreased acid output in the gastric secretion of zinc treated rats, increased the gastric mucosal barrier. Studies on the mechanism of action suggested the involvement of -SH groups in producing gastric antisecretory effect. Thus, zinc gluconate at > > 100 microM concentrations inhibited H(+)-ion transport which could be reversed by incorporating beta-mercaptoethanol in the secretory solution (luminal side). On the other hand, beta-mercaptoethanol added from the nutrient side showed no effect on the inhibition of H(+)-transport indicting that the implication of -SH groups may not be the sole factor. Zinc appeared to play a vital and multifaceted protective role in chemically induced gastric ulcer disorders.

Animals↗

The gluconate high affinity transport of GntI in Escherichia coli involves a multicomponent complex system.

Within the main system for gluconate utilization in E. coli, the gntT gene (located at the minute 76.4) that encodes a permease, is currently the only element involved in the high affinity transport. In this paper, the nucleotide sequence of the upstream region of this locus was determined. Two open reading frames of 729 bp (gntX) and 573 bp (gntY) were identified as additional gnt genes by complementation studies. Our observations suggest that these loci might conform an operon distinct of gntT under the control of the gntR gene product. Such operon encodes a gluconate periplasmic binding protein (GntX) and a putative membrane-bound protein (GntY). These products and the permease encoded by the gntT gene seem to conform a high-affinity complex transport system for gluconate. We suggest that this novel system could belong to the TRAP transporters.

Bacterial Proteins↗

Cochlear ototoxicity of chlorhexidine gluconate in cats.

Chlorhexidine gluconate is a derivative of chlorhexidine and is a popular disinfectant with a strong bactericidal action that is widely used for preoperative sterilization in ear surgery. The purpose of this investigation is to ascertain the potential ototoxicity of this agent. After topically applying chlorhexidine gluconate solutions to the middle ear cavities of 12 cats, we observed the excised cochleas using both scanning and transmission electron microscope studies. Either 0.05% or 2% chlorhexidine gluconate solutions were infused into the right ear of the test animal through one of two tubes chronically installed in the tympanic bullae. The left ears were utilized as controls and were infused with sterilized physiological saline. The solutions were administered once every other day for three separate infusions. Nine animals were decapitated 7 days after the third application, while the other three animals were sacrificed at 4 weeks. In the 2% chlorhexidine group, we found that hair cells in the organ of Corti had degenerated and had lost their hair bundles over a wide range. This pathology was more marked in the lower cochlear turns. In the animals sacrificed at 4 weeks, the injuries present seemed to have progressed. Even at a clinical concentration of 0.05%, chlorhexidine caused intracellular degeneration but with little surface damage. Our findings would suggest a cause of hearing loss when chlorhexidine is used clinically in the ear.

Administration, Topical↗

The effect of chlorhexidine gluconate irrigation on the root canal flora of freshly extracted necrotic teeth.

Forty freshly extracted teeth with necrotic pulps were endodontically treated under simulated clinical conditions. Bacteriologic samples were obtained before during, immediately after, and 24 hours after instrumentation, irrigation, and medication either with 0.2 percent chlorhexidine gluconate or with sterile saline. There was a highly significant (p less than 0.0049) reduction in microorganisms in the chlorhexidine-treated specimens after the instrumentation and irrigation procedures. Further significant reductions were noted after intracanal dressing of the teeth for 24 hours with the chlorhexidine gluconate solution. The teeth treated with saline also demonstrated a generalized decrease in flora after the biomechanical procedures. However, there was an absolute increase in numbers of microorganisms for 80 percent of the uniradicular and 50 percent of the multiradicular specimens when no intracanal, antimicrobial dressing was applied. It was concluded that chlorhexidine gluconate in a 0.2 percent solution can be an effective antimicrobial agent when used as an endodontic irrigating solution and that as an intracanal, interappointment dressing, chlorhexidine helps to further reduce bacteria remaining within the root canal system.

Bacteria↗

Efficacy evaluation of a 4% chlorhexidine gluconate as a full-body shower wash.

A 4% chlorhexidine gluconate (Xttrium Laboratories, Chicago, Ill.) was used in a shower bath application to evaluate its merits in reducing resident skin microorganisms. Five volunteers completed a 14-day microbial stabilization period, a 7-day baseline period, and a 5-day test period. Subjects followed a standard protocol, performed five shower washes and were sampled at both the abdominal and inguinal regions immediately after the shower wash as well as at 3 and 6 hours later on days 1, 2, and 5. Chlorhexidine gluconate provided significant microbial reductions from baseline, with greater reductions noted as the study progressed. Use of chlorhexidine gluconate in a shower wash application in conjunction with a preoperative skin-preparation procedure may reduce the probability of surgically associated infections.

Abdomen↗

Effect of gluconic acid as a secondary carbon source on non-growing L-lysine producers cells of Corynebacterium glutamicum. Purification and properties of 6-phosphogluconate dehydrogenase.

We studied the production of L-lysine in Corynebacterium glutamicum ATCC 21543 non growing cells obtained by nutrient limitation. Statistical analysis revealed significant differences in the L-lysine titers of glucose, gluconic acid or glucose-gluconic acid cultures. Higher L-lysine titer obtained in batch cultures with mixed carbon sources or gluconic acid alone were found to be associated with a high 6-phosphogluconate dehydrogenase activity (6PGDH, E.C.1.1.1.44). This enzyme is a pivotal enzyme within the hexose monophosphate pathway, and thus of importance for L-lysine production. 6PGDH was purified and characterized. The purified enzyme migrates as a single band on sodium dodecyl sulfate-polyacrylamide gel electrophoresis with a molecular mass of 52.5 kDa. The molecular mass of the native enzyme was estimated to be 120 kDa by molecular exclusion chromatography, thus suggesting a homodimeric structure. The amino terminal sequence shows a strong similarity (a match of 86% of the first 20 amino acid) to the 6PGDH from other microorganisms such as, E. coli and B. subtilis. The pI of the dimeric native enzyme and the optimum pH were 6.2 and 8.0, respectively. For the oxidative decarboxylation of 6-phosphogluconate, K(m) of 71 &mgr;M and 43 &mgr;M were obtained for 6-phosphogluconate and NADP(+), respectively.

Journal Article↗

Treatment of severe malaria in the United States with a continuous infusion of quinidine gluconate and exchange transfusion.

During the past decade the incidence of Plasmodium falciparum malaria in the United States has increased 10-fold. Treatment may be delayed because the therapy recommended for severe or complicated disease, intravenous quinine dihydrochloride, is available only from the Centers for Disease Control. We studied 17 patients who were treated for severe or complicated P. falciparum malaria in the United States between 1985 and 1987. Five patients were treated with a continuous infusion of quinidine gluconate, 10 with an exchange transfusion in addition to the continuous infusion of quinidine gluconate, and 2 with intermittently administered intravenous quinine dihydrochloride and an exchange transfusion. All 16 patients with P. falciparum malaria (1 patient had P. vivax malaria) had hyperparasitemia at the time of diagnosis (6 to 54 percent of the erythrocytes infected; median, 13 percent). Three patients with marked hyperparasitemia (54, 38, and 30 percent) and multiple other indicators of a poor prognosis, including advanced age, died. The 13 patients who completed their courses of quinidine with or without exchange transfusion had a parasitemia level of 1.1 percent or less 28 to 72 hours (mean, 44.4 hours) after the start of therapy. Side effects of quinidine treatment were observed in only two patients, one of whom had a serum quinidine concentration above the toxic level. We conclude that the continuous infusion of quinidine gluconate is well tolerated alone and with exchange transfusion and is effective in the treatment of severe and complicated malaria.

Adult↗

Effects of 0.2% chlorhexidine gluconate and amoxicillin plus clavulanic acid on the prevention of alveolar osteitis following mandibular third molar extractions.

OBJECTIVE: The purpose of this study was to evaluate the use of a 0.2% chlorhexidine gluconate and amoxicillin plus clavulanic acid combination as a prophylactic therapy for the prevention of alveolar osteitis after mandibular third molar extractions and to investigate adverse reactions to chlorhexidine. STUDY DESIGN: This randomized, placebo-controlled, parallel group study was conducted in a group of 177 subjects, from which 3 groups were formed. The first group (n = 62) received 0.2% chlorhexidine gluconate, the second group (n = 56) received a 0.2% chlorhexidine gluconate and amoxicillin plus clavulanic acid combination, and the third group (n = 59) received 0.09% sterile saline solution. All patients were recalled for the diagnosis of alveolar osteitis on the third and seventh postoperative days. RESULTS: When patients in the antibiotic group were compared with those in the other 2 groups, a significant reduction in alveolar osteitis was noted (P <.05). An alteration in taste, the bad taste of the solution, and staining of dentures and oral tissues were the major complaints about chlorhexidine. CONCLUSION: It would be more beneficial to use chlorhexidine solution with a beta-lactamase inhibitor-containing antibiotic to enhance its effectiveness for the prevention of alveolar osteitis.

Adult↗

Disinfection with 10% povidone-iodine versus 0.5% chlorhexidine gluconate in 70% isopropanol in the neonatal intensive care unit.

AIM: The finding that 10% povidone-iodine skin disinfectant may compromise thyroid function in premature infants prompted its replacement with 0.5% chlorhexidine gluconate solution in 70% isopropanol. The objective of this study was to compare the incidence rates of true infection and contamination associated with the use of these two disinfectants in the neonatal intensive care unit. METHODS: The study population comprised two cohorts of infants admitted to our neonatal intensive care unit: 1) in 1992-1993 when only 10% povidone-iodine was used as a skin disinfectant, and 2) in 1995-1996 when only 0.5% chlorhexidine gluconate solution in 70% isopropanol was used. A retrospective chart review was conducted to determine whether all documented positive blood, CSF and suprapubic aspirate cultures indicated true infection or contamination. True infection was defined as clinical symptoms and/or laboratory abnormalities suggestive of sepsis, with positive blood, CSF or suprapubic aspirate cultures. RESULTS: 1146 infants were admitted during the study periods, 507 during the first period and 639 during the second. In the early group, 17.6% of infants had major malformations, 72.0% were premature and 25.2% had weights of < 1500 g. Corresponding percentages for the latter group were 16.0%, 80.6% and 32.9%, respectively. No statistically significant differences were found between the two research periods in rate of infants with positive blood cultures, true infections, or contamination. CONCLUSION: The use of 0.5% chlorhexidine gluconate solution in 70% isopropanol as a skin disinfectant is justified in neonatal intensive care units because it is not associated with an increased incidence of infections as opposed to 10% povidone-iodine and is devoid of detrimental effects.

Anti-Infective Agents, Local↗