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Biomedical subjects

B Bachmann

Publications and source records attributed to B Bachmann.

At least 19 recordsLinked to original sources

A special reactor design for investigations of mixing time effects in a scaled-down industrial L-lysine fed-batch fermentation process

A specially designed model reactor based on a 42-L laboratory fermentor was equipped with six stirrers (Rushton turbines) and five cylindrical disks. In this model reactor, the mixing time, Theta(90), turned out to be 13 times longer compared with the 42-L standard laboratory fermentor fitted with two Rushton turbines and four wall-fixed longitudinal baffles. To prove the suitability of the model reactor for scaledown studies of mixing-time-dependent processes, parallel exponential fed-batch cultivations were carried out with the leucine-auxotrophic strain, Corynebacterium glutamicum DSM 5715, serving as a microbial test system. L‐Leucine, the process-limiting substrate, was fed onto the liquid surface of both reactors. Cultivations were conducted using the same inoculum material and equal oxygen supply. The model reactor showed reduced sugar consumption (-14%), reduced ammonium consumption (-19%), and reduced biomass formation (-7%), which resulted in a decrease in L-lysine formation (-12%). These findings were reflected in less specific enzyme activity, which was determined for citrate synthase (CS), phosphoenolpyruvate carboxylase (PEP-C), and aspartate kinase (AK). The reduced specific activity of CS correlated with lower CO(2) evolution (-36%) during cultivation. The model reactor represents a valuable tool to simulate the conditions of poor mixing and inhomogeneous substrate distribution in bioreactors of industrial scale. Copyright 1999 John Wiley & Sons, Inc.

Journal Article↗

Determination of trehalose by flow injection analysis using immobilized trehalase.

A new method for the determination of trehalose by flow injection analysis (FIA) is described. The basic principle is the hydrolysis of the disaccharide trehalose into its monomer d-glucose by trehalase, a periplasmic enzyme of Escherichia coli. d-glucose is quantified spectrophotometrically after reaction with hexokinase and glucose-6-phosphate dehydrogenase. Trehalase is prepared by osmotic shock from a recombinant E. coli strain and precipitated with ammonium sulfate. The enzyme is immobilized on VA-Epoxy Biosynth from Riedel-de-Haën. The immobilization rate is about 60%. The FIA signals show a nonlinear dependence on the trehalose concentration. The resulting curve corresponds to a second-order polynomial that serves as a calibration function for test samples. Immobilized trehalase was used during a period of 4 months without any loss of suitability. Several samples of fermentation broth were tested. The results are verified by HPLC. Within an interval of 2 to 10 g/L trehalose the recovery is about 100-120% with a precision of 7% (coefficient of variation).

Autoanalysis↗

7-alkylidenecephalosporin esters as inhibitors of human leukocyte elastase.

A series of 7-alkylidenecephalosporins and 7-vinylidenecephalosporins, as their benzhydryl esters, have been tested as inhibitors of both porcine pancreatic elastase and human leukocyte elastase. Selected 7-alkylidenecephalosporin esters are found to be potent inhibitors of HLE. One category of new inhibitors is the 7-(haloalkylidene)cephalosporins. In contrast to previously reported cephalosporin-based elastase inhibitors, these haloalkylidene cephems show optimum inhibitory activity as sulfides, rather than as sulfones. They are efficient and irreversible inhibitors. A second class of active compounds is represented by the benzhydryl ester 7-(cyanomethylidene)cephalosporin sulfone. In contrast to the activity of these new inhibitors, the benzhydryl ester of the mechanism-based beta-lactamase inhibitor, 7-[(2'-pyridyl)methylidene]-cephalosporin sulfone showed little inhibitory activity as an elastase inhibitor. 7-Vinylidenecephalosporins were also relatively poor inhibitors, although the terminally unsubstituted allene sulfide showed activity as an inhibitor of PPE. A modeling analysis suggests the 7-alkylidene substituents can be readily accommodated in the S1 pocket. A potential mechanism of inhibition is proposed.

Animals↗

Virus inactivation of blood products by phenothiazine dyes and light.

Methylene blue (MB) and its derivatives azure A, B, C and thionine are photoactive and, in principle, are suitable for photodynamic virus inactivation of blood and blood products, such as therapeutic plasma. Methylene blue was selected for plasma decontamination because it is being clinically used and because of its known toxicological and other properties. The standard procedure for photodynamic treatment of single units of fresh plasma involves illumination with visible light at an MB concentration of 1 microM. Polymerase chain reaction analysis revealed that, in addition to model viruses, the bloodborne viruses hepatitis B virus, hepatitis C virus, human immune deficiency virus-1 and probably also the nonenveloped parvovirus B19 are sensitive to MB/light treatment. The procedure is further improved when the fluorescent tubes routinely used for illumination are replaced by more intense light sources, e.g. light-emitting diodes or low-pressure sodium lamps. Surprisingly, the improved virus kill is accompanied by reduced damage to plasma proteins.

Blood-Borne Pathogens↗

Bupivacaine plasma concentrations during continuous epidural anesthesia in infants and children.

Venous bupivacaine plasma concentrations were measured in six neonates and infants aged 4 days to 3.9 mo (mean, 2.1 mo) and 10 infants and children aged 9 mo to 6 yr (mean, 3.1 yr) after administration of an initial bolus of 0.5 mL/kg bupivacaine 0.25%, followed by a continuous infusion of local anesthetic (0.25 mL.kg-1.h-1) over a period of 4 h (first hour: bupivacaine 0.25%, then reduced to 0.125%). Plasma concentrations of local anesthetic measured at 180 min and 300 min after beginning of bupivacaine administration were significantly higher in younger infants when compared to older infants and children (180 min: 0.67 +/- 0.24 micrograms/mL [0.25-0.97] vs 0.27 +/- 0.11 micrograms/mL [0.19-0.55], P < 0.01; 300 min: 0.86 +/- 0.36 micrograms/mL [0.35-1.25] vs 0.34 +/- 0.12 micrograms/mL [0.18-0.57], P < 0.01). The results of our study show that despite applying the same dosage of epidural bupivacaine significantly higher plasma concentrations were seen after short periods of continuous infusion in infants up to 4 mo than in children older than 9 mo.

Age Factors↗

Synthesis and biological activity of 7-alkylidenecephems.

Several 7-alkylidenecephalosporins were synthesized and biologically evaluated as beta-lactamase inhibitors. The three beta-lactamase enzymes used in this study included two type C beta-lactamases, derived from Enterobacter cloacae P99 and E. cloacae SC12368, and one type A beta-lactamase, derived from Escherichia coli WC3310. Of the cephalosporins prepared, compound 7e, the sodium salt of 7-[(Z)-(2'-pyridyl)methylene]cephalosporanic acid sulfone, was found to have excellent inhibitory properties against both type C enzymes. Also, compound 7f, the sodium salt of 7-[(Z)-(tert-butoxycarbonyl)methylene]cephalosporanic acid sulfone showed high activity as an inhibitor of the type A enzyme. The inhibition kinetics of 7e were further explored. The IC50 value of 7e indicated that this compound was approximately 20-fold more active than tazobactam against the enzyme derived from E. cloacae P99 and 167-fold more active than tazobactam against the enzyme derived from E. cloacae SC12368. A plot of enzymatic activity vs incubation time with stoichiometric amounts of inhibitor reveals a rapid deactivation of the enzyme followed by an extremely slow reactivation. 7e exhibited a second-order rate constant of k3' = 5.3 x 10(6) L/mol.min, and a partition ratio of approximately 20:1 inhibitor:enzyme was determined for this inhibitor. After separation of excess inhibitor with Sephadex filtration, a rate constant of enzyme reactivation was measured at kreactiv = 1.0 x 10(-3) s-1. Following 24 h of incubation of enzyme with a large excess of inhibitor and sephadex filtration to remove excess inhibitor, the enzyme was able to recover only 43% of its original activity, indicating an irreversible component to the inhibition. Potential mechanisms of inhibition for both 7e and 7f are suggested.

Anti-Bacterial Agents↗

Target structures for HIV-1 inactivation by methylene blue and light.

In a photodynamic virus inactivation procedure for human fresh frozen plasma the plasma is exposed to visible light in the presence of 1 microM methylene blue. This procedure is known to inactivate HIV-1 by at least 10(6.32) TCID50/ml within 10 minutes. To elucidate the mechanism of photodynamic inactivation of HIV-1 by methylene blue/light treatment, reverse transcriptase (RT), the HIV-1 associated protein p24, and viral RNA were examined. In the dark, methylene blue up to 10 microM has no inhibitory effect on recombinant RT. In the presence of light, recombinant RT inactivation was dependent on illumination time and the concentration of methylene blue. After photoinactivation of the whole virus by methylene blue/light treatment, RT activity was also almost completely inhibited. Simultaneously, it was found by Western blotting that HIV-1 p24 and gp120 are altered in size, possibly due to protein cross-linking. In addition, it was shown by polymerase chain reaction (PCR) inhibition assay that HIV-1 inactivation leads to destruction of its RNA. In summary, methylene blue/light treatment acts on HIV-1 at different target sites: the envelope and core proteins, and the inner core structures RNA and RT.

Antiviral Agents↗

The effects of epidural and intravenous lidocaine on somatosensory evoked potentials after stimulation of the posterior tibial nerve.

The measurement of somatosensory evoked potentials (SEPs) after stimulation of the posterior tibial nerve (PTN-SEPs) has been proposed as an objective indicator of the quality of lumbar epidural block. It is unclear whether peak latency increases after epidural application of local anesthetics may be due in part to systemic effects of the drug absorbed from the epidural space. In this clinical study, we compared PTN-SEPs after intravenous and epidural administration of lidocaine to those of a control group who did not receive lidocaine. Plasma concentrations of lidocaine remained within expected ranges for epidural and intravenous administration. No subjects developed signs for overdose or toxicity. After epidural application of 2% lidocaine, mean latencies of peaks P1, N1, and P2 increased significantly in comparison to baseline values. In 3 of 10 patients, latency changes were not observed. Intravenous lidocaine did not produce statistically significant changes in latencies, although a trend toward increasing latencies appeared to be present. In the control group without lidocaine, no statistically significant changes occurred during the 1-h study period. No correlation was found between peak latency changes and plasma concentrations of lidocaine. We conclude that latency increases observed after epidural application of lidocaine are due primarily to local, not systemic, effects of the local anesthetic.

Absorption↗

Effectiveness of coronary heart disease risk management in high-risk patients.

The purpose of this study was to investigate the efficacy of the patient-based coronary heart disease (CHD) risk management strategy in general practice by analyzing patients who were sent to a specialized center as refractory to conventional treatment. Of the 452 patients studied, 152 were excluded because of secondary hyperlipidemia. The effects of a food protocol-monitored dietary/drug treatment on lipid profiles and CHD risk indices and the compliance to this approach were monitored for 12 months in 300 of 452 patients. CHD risk classification was performed according to NIH criteria by the referring physicians, but secondary hyperlipidemia was not identified and treated appropriately. Physicians did not fully utilize dietary and drug treatment and referred the patients to a specialized center too early. The initiated food protocol-controlled treatment was more efficient than pretreatment in general practice, with a compliance of 80.3% by patients selected according to their CHD risk. Long-term CHD risk reduction was persistent for 12 months in compliant patients; however, a cumulative dropout rate of 43% after 6 months and of 68% after 12 months was noted. Statistical analysis failed to reveal consistent prognostic factors of long-term compliance.

Apolipoproteins A↗

[A possible risk for geriatric risk patients caused by intraoperative disorder of cerebral energy utilization?].

OBJECTIVE: This study was carried out on cerebrospinal fluid (CSF) to investigate the perioperative course of certain ischaemic markers, namely neurone-specific enolase (NSE), creatine kinase (CK-BB), hypoxanthine, and lactate in order to identify a disturbed cerebral energy utilisation which could be responsible for the development of temporary mental dysfunctions. Those dysfunctions are characterised by preserved memory content and perception, but the coordination and association of these functions are disturbed. Typical clinical signs are motor restlessness, disordered emotions, and symptoms of dementia. Little is known about the aetiology of those symptoms, but they are most likely due to various events, such as direct drug effects, the extent of surgical trauma, sensorial deprivation, and disturbed perfusion. METHODS: Eight orthopaedic patients (ASA III or IV) scheduled for removal of their total hip replacement were anaesthetised by catheter-spinal anaesthesia (CSA) for pain relief in combination with standardised, modified neuroleptanalgesia (NLA). At six defined times (15 hours preoperatively, immediately before and after surgery and 6, 24, and 36 hours postoperatively) CSF samples were drawn and the ischaemic markers were determined by means of radioimmunoassay (NSE), electrophoresis (CK-BB), photometry (lactate), and high-pressure liquid chromatography (hypoxanthine). The release of ischaemic markers into CSF correlates linear with the extent of ischaemic brain damage. RESULTS: Mean concentrations of the following ischaemic markers increased in all patients intraoperatively: NSE from 12.3 ng/ml to 13.4 ng/ml, hypoxanthine from 1.86 mumol/l to 3.73 mumol/l, and lactate from 1.4 mmol/l to 2.0 mmol/l respectively, all of which returned to normal within 36 hours. The CK-BB concentrations were all within normal values and not affected by the operation during this investigation. CONCLUSION: Although no clinical signs of temporary mental dysfunction have been observed, the results indicate that in CSF ischaemic markers temporarily undergo certain changes in their concentrations during the removal of total hip replacements in elderly patients. These changes are reason for assuming that risk patients may suffer a temporary disturbed cerebral energy utilisation intraoperatively, even if stable clinical and cardiovascular conditions prevail under anaesthesia. Such a temporary ischaemic penumbra might be responsible for the postoperative development of temporary mental dysfunctions.

Aged↗

Variability of the env gene in cynomolgus macaques persistently infected with human immunodeficiency virus type 2 strain ben.

The sequence variability of distinct regions of the proviral env gene of human immunodeficiency virus type 2 strain ben (HIV-2ben) isolated sequentially over 3 to 4 years from six experimentally infected macaques was studied. The regions investigated were homologous to the V1, V2, V3, V4, V5, and V7 hypervariable regions identified in the env genes of HIV-1 and simian immunodeficiency virus SIVmac, respectively. In contrast to findings with HIV-1 and SIVmac, the V1- and V2-homologous regions were found to be highly conserved during the course of the HIV-2ben infection in macaques. The V3-homologous region showed a degree of variation comparable to that of HIV-1 but not of SIV. In the V4-, V5-, and V7-homologous regions, mutation hot spots were detected in most reisolates of the infected monkeys. Most of these mutations occurred during the first 10 weeks after infection. After 50 weeks, new mutations were rarely detected. At most mutation sites, a dynamic equilibrium between the mutated viral isotype and the infecting predominant wild type was present. This equilibrium might prevent an accumulation of mutations in isolates later in the course of infection.

Amino Acid Sequence↗

Identification of crossovers in Wilson disease families as reference points for a genetic localization of the gene.

Wilson disease (WD) is an autosomal recessive disorder of copper metabolism. A minimum recombinant analysis using D13S22, ESD, RB1, D13S31, D13S55, D13S26, D13S39, and D13S12, all localized at 13q14-q22, has been carried out in 20 WD families of Northwest-European origin. No inconsistencies have been observed with respect to locus order or location of the WD locus (WND) compared with previous linkage studies. D13S31 was mapped as the closest marker proximal to WND, whereas D13S55 and D13S26 were mapped as the closest markers distal to WND. We have identified a crossover between WND and D13S31 in one family and a crossover between WND and D13S55 in another. These crossover sites can be used as reference points for new chromosome 13q14-q21 markers, and are therefore important for a more accurate mapping of the WD locus.

Chromosome Mapping↗

[Fetal methemoglobinemia caused by prilocaine--is use of prilocaine for pudendal block still justified?].

17 women received 2 x 10 ml prilocaine 1% as a pudendal block sub partu. At delivery, the foetomaternal distribution ratio of the local anaesthetic was evaluated and the development of Met-Hb-concentration in the neonate was measured up to six hours post partum. The Met-Hb-concentration in the neonate was relatively low with a maximum of 1.8% after two hours, followed by a steady decline. A probable explanation for the Met-Hb-concentration could be the unexpected low foetomaternal ratio of distribution (0.5) and the increased renal elimination of the amide-type local anaesthetic in the neonate, respectively. According to these results, no contraindication for prilocaine in pudendal block is indicated.

Administration, Intravaginal↗

Genetic and biochemical analysis of the aspartokinase from Corynebacterium glutamicum.

The lysC/asd gene cluster of Corynebacterium glutamicum ATCC 13032 was cloned and sequenced. The lysC locus coding for aspartokinase consists of two in-frame overlapping genes, lysC alpha encoding a protein of 421 amino acids (Mr 44,300) and lysC beta encoding a protein of 172 amino acids (Mr 18,600). The C. glutamicum aspartokinase was purified and found to contain two proteins of Mr 47,000 and Mr 18,000. A C. glutamicum mutant expressing a feedback-resistant aspartokinase was shown to be changed in a single base pair of the lysC beta gene, leading to an amino acid exchange in the beta-subunit of the aspartokinase. In addition, the identified mutation was found to be responsible for the enhanced expression of the asd gene located downstream of lysC.

Amino Acid Sequence↗