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Biomedical subjects

C Koch

Publications and source records attributed to C Koch.

At least 163 records · Page 9Linked to original sources

On the relationship between synaptic input and spike output jitter in individual neurons.

What is the relationship between the temporal jitter in the arrival times of individual synaptic inputs to a neuron and the resultant jitter in its output spike? We report that the rise time of firing rates of cells in striate and extrastriate visual cortex in the macaque monkey remain equally sharp at different stages of processing. Furthermore, as observed by others, multiunit recordings from single units in the primate frontal lobe reveal a strong peak in their cross-correlation in the 10-150 msec range with very small temporal jitter (on the order of 1 msec). We explain these results using numerical models to study the relationship between the temporal jitter in excitatory and inhibitory synaptic input and the variability in the spike output timing in integrate-and-fire units and in a biophysically and anatomically detailed model of a cortical pyramidal cell. We conclude that under physiological circumstances, the standard deviation in the output jitter is linearly related to the standard deviation in the input jitter, with a constant of less than one. Thus, the timing jitter in successive layers of such neurons will converge to a small value dictated by the jitter in axonal propagation times.

Action Potentials↗

A non-denaturing enzyme linked immunosorbent assay with protein preadsorbed onto aluminum hydroxide.

A method has been developed which prevents denaturation of proteins used for coating of plastic surfaces in enzyme linked immunosorbent assays (ELISA). The system takes advantage of the use of aluminum hydroxide (Al(OH)3) as an adsorbent for proteins. A model protein has been analyzed. and monoclonal antibodies specific for either the native form or the denatured form of the protein were used to monitor the extent of denaturation. Adsorption of the proteins to Al(OH)3 in carbonate buffer, pH 9.3, before coating the ELISA plate abolished the denaturation otherwise observed after direct adsorption of protein to plastic surfaces. The protection against denaturation was dependent on the buffer system and was not observed when phosphate buffers were used, due to elution of protein from Al(OH)3 or lack of binding to Al(OH)3 in the presence of phosphate. There is evidence that protein adsorbed onto the Al(OH)3 is required for binding of Al(OH)3 onto the plastic surface. This system may be useful in assay systems where discrimination between the native and denatured forms of proteins is important.

Adsorption↗

Antibiotic treatment of initial colonization with Pseudomonas aeruginosa postpones chronic infection and prevents deterioration of pulmonary function in cystic fibrosis.

Chronic pulmonary infection with Pseudomonas aeruginosa (PA) develops in most patients with cystic fibrosis (CF) and is associated with a poor prognosis. Much effort has been directed toward treating the chronic infection, but it is almost impossible to eradicate it once established; therefore, prevention is preferable. Since 1989 CF patients at the Danish CF Center in Copenhagen have been treated with an intensive three-step-protocol consisting of colistin inhalations and oral ciprofloxacin at the time of initial PA colonization. This study compares 48 patients treated according to this intensive protocol with 43 historic controls. The study was carried out over 44 months and included 218 patient-years. Only 16% of the treated patients developed chronic PA infection after 3 1/2 years compared with 72% of the control patients (Kaplan Meier estimate, P < 0.005, log rank test). This indicates that aggressive treatment prevented or delayed chronic PA infection in 78% of the patients for 3 1/2 years. Furthermore, aggressive treatment maintained or increased pulmonary function (forced vital capacity and forced expiratory volume in 1 second in percent of predicted values) during the year after inclusion compared with the control group, in which pulmonary function declined (P < 0.01, Mann-Whitney test). Although some of the treated patients eventually developed chronic PA infection, these patients had significantly better pulmonary function at the onset of chronic PA infection compared with control patients (P < 0.001, Mann-Whitney test). When the different steps in the intensive three-step-protocol were analyzed, there was a trend suggesting that 3 months of high-dose treatment with colistin inhalation and oral ciprofloxacin produced the best results in terms of postponement or prevention of chronic PA infection (P < 0.05).

Adolescent↗

International practice patterns by age and severity of lung disease in cystic fibrosis: data from the Epidemiologic Registry of Cystic Fibrosis (ERCF).

The Epidemiologic Registry of Cystic Fibrosis provides clinical profiles for more than 6,800 patients and descriptions of practice patterns across eight European countries. Preliminary cross-sectional analysis has been performed by age and pulmonary function as an assessment of disease severity. In general, pulmonary treatments including inhaled bronchodilators and rhDNase increased as lung disease became more severe. Use of a number of treatments, including mucolytic agents and inhaled corticosteroids, varied markedly from country to country. Several widely used therapies are not yet supported by controlled clinical trials, particularly in patients under 6 years of age. Nutritional intervention was more common in patients with advanced lung disease regardless of age. Patients with nasal polyps had less severe lung disease at each age than patients without polyps. It is clear that studies of early interventions are needed to determine the optimal types of treatments and the ages at which to begin treatment.

Adolescent↗

Stimulus and response factors affecting the development of behavioral sensitization to apomorphine.

The present study was designed to assess the role of stimulus and response factors in the context-dependency of behavioral sensitization to the direct dopamine agonist apomorphine. In two experiments, male Wistar rats were given repeated injections of the direct dopamine agonist, apomorphine (5 mg/kg, s.c.), or vehicle at 24- to 72-h intervals and tested for locomotor activity for 30 min in either an openfield activity drum or a running wheel. In experiment 1, after eight activity sessions in either the activity drum or running wheel, one-half of the rats in each drug condition (apomorphine or vehicle) were tested in the alternate activity test environment. In both activity test environments, apomorphine produced progressively greater levels of locomotor activity with repeated treatment (i.e., sensitization). Moreover, sensitization to apomorphine transferred completely across test environments. That is, rats given apomorphine associated with one test environment (e.g., wheel) displayed equivalent sensitization when tested in the alternate environment (e.g., drum). Thus, changing external stimulus cues associated with repeated drug exposure did not affect the expression of sensitization. In experiment 2, rats were given either apomorphine or vehicle daily and tested for activity in a running wheel. For one-half the rats in each drug condition, the running wheel was free to move, but for the remainder the wheel was immobilized. After nine training sessions, all rats were given an apomorphine challenge injection and tested in a mobile wheel. After the challenge injection of apomorphine, rats previously treated with apomorphine and trained in the mobile wheel were significantly more active than rats previously treated with vehicle. In contrast, rats given equivalent apomorphine treatments and trained in the immobile wheel did not differ in activity from rats previously given only vehicle. Since the external stimulus cues associated with drug exposure for the mobile and immobile wheel groups were the same, this finding suggests that environmental factors affecting response expression are critical to the development of behavioral sensitization to apomorphine.

Animals↗

Spatial vision thresholds in the near absence of attention.

It is well known that attention increases the discriminability of some types of spatial information. To ascertain more specifically which types of spatial information benefit from attention, we have measured spatial vision thresholds both in the presence and in the near absence of attention. To obtain near absence of attention, we induce subjects to focus attention elsewhere in the display by means of a suitably demanding concurrent visual task. We measure contrast and orientation thresholds for sine-wave gratings, as well uni- and bidirectional offset thresholds for vernier targets. The results suggest that attention selectively lowers some thresholds but not others: orientation thresholds are far more affected than contrast thresholds, and bidirectional vernier thresholds are far more affected than unidirectional thresholds.

Attention↗

Soluble haemoglobin is a marker of recent Plasmodium falciparum infections.

Monoclonal antibodies (Mab) were raised against haemoglobin (Hb) associated with Plasmodium falciparum protein and used to develop an ELISA, measuring circulating levels of released Hb. This assay was evaluated in different malaria patients in parallel with ELISA assays for C-reactive protein (CRP) and haptoglobin. Levels of Hb were negatively associated with levels of haptoglobin. Increased levels of serum Hb and CRP and decreased levels of haptoglobin were seen in Danish malaria patients. Consecutive studies showed that increased Hb levels were detectable 3-7 days after initiation of treatment probably because of drug induced destruction of infected erythrocytes. Increased levels of CRP were measured 0-3 days after initiation of treatment. The Hb assay was used in an epidemiological study of malaria in an area of Sudan with unstable malaria transmission. The proportion of Sudanese adults with detectable soluble Hb was higher in the rainy season with malaria transmission compared to the dry season. Hb levels in the rainy season were negatively associated with levels of haptoglobin. Most adults had increased levels of soluble Hb and decreased levels of haptoglobin 7 and 30 days after their treatment of P. falciparum malaria compared to the levels during acute disease. Thus, both soluble Hb and haptoglobin appear to be markers of recent P. falciparum infections. Very high levels of CRP protein were measured in some of the malaria patients at the day of treatment while lower levels were recorded 7 and 30 days after treatment. Soluble Hb levels were associated with malariometric parameters in a similar fashion to haptoglobin. The new Mab-based assay for measuring soluble Hb in the peripheral blood of malaria patients may be useful for future epidemiological studies of malaria.

Adult↗

Epidemiology of Pseudomonas aeruginosa in cystic fibrosis and the possible role of contamination by dental equipment.

Cystic fibrosis (CF) patients often suffer from Pseudomonas aeruginosa lung infection yet the source of this organism is not known. In order to determine whether CF patients might be contaminated with P. aeruginosa from dental equipment, a total of 103 water samples from 25 dental sessions in Frederiksberg Municipal Oral Health Care Service were examined. Three samples (2.9%) were positive for P. aeruginosa. Three hundred and twenty-seven water samples from 82 dental sessions from various other Municipal Oral Health Services in Denmark, attended by CF patients, were also examined. Eighteen of 327 samples (5.5%) from nine sessions (11%) were positive for P. aeruginosa. In one case, genotypically identical (RFLP, pulsed-field gel electrophoresis) P. aeruginosa strains were found both in water from the dental equipment and in the CF patients sputum. This indicates a small risk for acquiring P. aeruginosa from dental sessions, which is however equal to the yearly 'natural background' incidence (1-2%) of acquisition of P. aeruginosa in our CF centre.

Case-Control Studies↗

Shock-wave measurement using a calibrated interferometric fiber-tip sensor.

The results of shock-wave measurements using a calibrated fiber-tip sensor based on a Michelson interferometer are presented. A transfer function, obtained by an independent experiment that describes the properties of the sensor system, was used to correct the measured shock-wave data in the Fourier frequency domain. The phase of the transfer function was determined from its amplitude by a fitting procedure using minimum-phase terms. As an example of application, the acoustic output field of an electromagnetic lithotriptor was investigated, and the shock-wave source was reliably characterized. The measured data provide a basis for estimating the hazard to which a patient is exposed during shock-wave treatment and for optimizing a lithotriptor system to produce a sharply localized and effective acoustic field.

Algorithms↗

Inhibition of LPS and Plasmodium falciparum induced cytokine secretion by pentoxifylline and two analogues.

Pentoxifylline and the two analogues HWA138 and HWA448, at concentrations exceeding 60 micrograms/ml, inhibited malaria antigen or lipopolysaccharide (LPS) induced TNF-alpha and IL-1 alpha secretion, but not IL-6 secretion, from human peripheral blood mononuclear cells in vitro. HWA448 had lower inhibitory activity in vitro than pentoxifylline and HWA138. A small enhancement of cytokine secretion was induced by pentoxifylline and the two analogues at low concentrations. The drugs did not affect cell viability. Pentoxifylline, HWA138 and HWA448 also inhibited LPS induced TNF production in vivo in female CF1xBalb/c mice. The drugs were inhibitory at 0.5-1 mg per mouse when mixed with LPS, and 1 mg per mouse of the drugs was inhibitory when injected 1 h before LPS challenge. HWA448 had similar inhibitory activities in vivo compared to pentoxifylline and HWA138, possibly because of the longer serum half-life of HWA448. The pentoxifylline analogues may have lower toxicity than pentoxifylline itself and may therefore be useful in future treatment of diseases induced by endotoxic substances.

Adult↗

The anesthetic effect of dexmedetomidine does not adhere to the Meyer-Overton rule but is reversed by hydrostatic pressure.

To test the hypothesis that the anesthetic action of dexmedetomidine is similar to that of other anesthetics, dexmedetomidine was studied using two classical descriptors of anesthesia, the Meyer-Overton correlation and the pressure reversal of anesthesia. The anesthetic potency of dexmedetomidine and its isomer, levomedetomidine, were determined in Xenopus laevis tadpoles. Anesthesia was defined as loss of righting reflex. Octanol/water partition coefficients were determined using ultraviolet absorbance spectrophotometry. Pressure experiments were performed at pressures ranging from 1 to 91 atmospheres absolute (ata). A concentration-response curve was obtained with a half-maximum effective concentration (EC50) of dexmedetomidine of 7.1 +/- 1.1 microM (mean +/- SEM), whereas levomedetomidine showed an EC50 of 59.3 +/- 5.5 microM. The octanol/water partition coefficient was 206 +/- 49. With increasing pressure, the EC50 of dexmedetomidine increased linearly up to a value of 23.9 +/- 3.2 microM at 91 ata. Correlation of lipid solubility and anesthetic potency demonstrates that dexmedetomidine does not adhere to the Meyer-Overton rule. However, the anesthetic action of dexmedetomidine is diminished by increased pressures, thus demonstrating a similar effect compared with other general anesthetics.

Anesthetics↗

Phylogenetic evidence for the taxonomic heterogeneity of Photorhabdus luminescens.

The sequences of the 16S rRNA gene of 40 strains of bacterial symbionts isolated from the nematodes Heterorhabditis spp. and seven bacterial symbionts of the nematodes Steinernema spp. which were isolated from different geographical areas, as well as the type strain of Xenorhabdus japonicus, were determined and compared to each other and to the sequences of several reference strains of members of the Enterobacteriaceae. The data confirmed the separate status of the two genera of symbionts of entomopathogenic rhabditid nematodes. The symbionts of Heterorhabditis spp. clustered with the type strain of Photorhabdus luminescens, while the symbionts of Steinernema spp. grouped with Xenorhabdus species. X. japonicus clustered with the other Xenorhabdus species. Phylogenetic analysis of 15 almost complete 16S ribosomal DNA (rDNA) sequences of the Heterorhabditis symbionts indicated that there were several subclusters. The properties correlated with these subclusters are not yet apparent, although there may be some geographical and ecological correlations. For example, among the nematode-symbiotic bacteria, the members of subclusters I and III are from southeastern and midwestern North America, respectively, while the members of subclusters II and IV are primarily from Europe and Australia, respectively. The nonsymbiotic strains of P. luminescens form a highly homologous subcluster by themselves. The results of DNA-DNA hybridization studies performed with a few selected strains of five of the 16S rDNA subclusters support the existence of several genospecies within P. luminescens.

Animals↗

Production and characterization of mouse monoclonal antibodies against Afipia felis.

A series of 10 monoclonal antibodies reacting with Afipia felis antigens were selected from mice immunized with live organisms of the reference strain ATCC 53690. Immunoblotting against SDS-PAGE-separated A felis sonicate allowed the antibodies to be classified into three groups: 1) 168-4, -6, -7 and -10 reacted with a 53 kDa antigen, 2) 168-1, -3 and -9 reacted with both 53 kDa and 60 kDa antigens, and 3) 168-2, -5 and -9 reacted with other antigens. Antibodies of group 1 did not cross-react with other Afipia species or 36 unrelated bacteria, whereas those of groups 2 and 3 reacted with other Afipia species and some unrelated bacteria. Immunoblots of crossed immunoelectrophoretic patterns of A. felis sonicate against rabbit antiserum showed that antibodies of groups 1 and 2 bound to the same precipitin arcs. Antibodies of group 1 reacted with a species-specific epitope on the 53 kDa antigen, while those of group 2 reacted with other epitopes shared by the 53 kDa and 60 kDa antigens. The binding of antibodies of group 1 to A. felis sonicate was inhibited by post-infection rabbit serum, whereas no inhibition was observed for antibodies of group 2. The species-specific epitope of the 53 kDa antigen and the early appearance of antibodies against this epitope after infection suggest that this antigen can be used in a serodiagnostic test for A. felis infection.

Animals↗

Identification of Afipia felis antigens in culture medium: reaction with human sera.

Fourteen protein antigens were identified on SDS-PAGE of Afipia felis culture supernatant. Immunoblotting against 10 monoclonal antibodies obtained from mice infected with live A. felis showed that 4 antibodies reacted with a 56 kDa band and 3 with both 56 kDa and 62 kDa bands. Compared with A. felis sonicate, the reacting proteins in culture supernatant showed an increase in molecular mass of 2-3 kDa, suggesting that they were more glycosylated. Purified antigen obtained by affinity chromatography of culture supernatant on the seven immobilized antibodies was tested against antibodies reacting with the 56 kDa and 62 kDa bands. All eluates contained both components, suggesting that the antibodies were directed against different epitopes of a double antigen held together during the affinity chromatography but cleaved by reduction and SDS-PAGE. The molecular size of the uncleaved protein in culture supernatant was determined by size-exclusion chromatography as > 1000 kDa. Testing of pre- and post-infection rabbit sera in immunoblotting against culture supernatant demonstrated that the 56 kDa and 62 kDa components gave the most prominent specific reactions with post-infection sera. One of fifty human sera submitted for testing for cat-scratch disease and 1 of 50 sera from healthy blood donors reacted with several bands in A. felis culture supernatant, including the 56 kDa and 62 kDa bands.

Animals↗

Evolution and probable transmission of intersubtype recombinant human immunodeficiency virus type 1 in a Zambian couple.

The extraordinary genetic diversity of human immunodeficiency virus type 1 (HIV-1) results from the introduction of mutations by an error-prone reverse transcriptase and from recombination of the two RNA genomes packaged in the virion during the synthesis of proviral DNA. The occurrence of multiple, genetically distant HIV-1 subtypes and their geographic intermixing set up conditions for dramatic, rather than gradual, changes in genotype whenever genomes from different subtypes are copackaged in virions. Here we describe, for the first time, the sequential generation of multiple different, but related, intersubtype HIV-1 recombinants within an infected individual. Full-length gag and env genes were recovered directly from peripheral blood mononuclear cells or from primary virus cultures, using serial blood samples from a Zambian woman and a sample from her spouse. DNA sequencing and phylogenetic analysis established that two different A/C recombinant forms of HIV-1 predominated at two time points in the woman. A related but distinct recombinant HIV-1 was recovered from her spouse. Intersubtype recombination apparently played a central role in the evolution of HIV-1 in this couple and may contribute substantially to the rapid emergence of HIV-1 variants whenever mixed-subtype HIV-1 infections occur.

Base Sequence↗

Flow resistance of individual neutrophils in coronary artery disease: decreased pore transit times in acute myocardial infarction.

OBJECTIVE: To investigate single neutrophil flow resistance in coronary artery disease, including myocardial infarction before initiation of reperfusion therapy. DESIGN: Neutrophil flow resistance was measured in 93 subjects in five groups: (group 1) 28 patients within 12 hours after the onset of myocardial infarction, before reperfusion therapy; (group 2) 18 with unstable angina; (group 3) 13 with stable angina; (group 4) 13 age matched patients without coronary disease, and (group 5) 21 healthy volunteers. MAIN PARAMETERS: Single neutrophil transit times through 8 microns oligopore filters determined with a modified cell transit analyser. RESULTS: Leucocyte count (10(9)/l) was increased in coronary disease, especially in myocardial infarction and unstable angina (mean and 95% confidence intervals for groups 1 to 5: 12.6 (11.0 to 14.2), 11.3 (8.5 to 14.1), 8.5 (7.4 to 9.6), 8.0 (6.0 to 10.0), 7.0 (6.1 to 7.9)). Polymorphonuclear granulocyte (PMN) flow resistance correlated negatively with white blood cell (WBC) count and was significantly decreased in coronary artery disease (CAD), especially in myocardial infarction; mean transit times (ms) for groups 1 to 5 were: 13.6 (11.8 to 15.4), 16.9 (13.9 to 19.0), 16.9 (12.8 to 21.0), 22.0 (19.6 to 24.4), and 18.6 (15.7 to 21.5). CONCLUSION: Neutrophil flow resistance was decreased in CAD, especially in myocardial infarction before reperfusion therapy. In contrast to previous findings in reperfused myocardial infarction, the present study showed that stiffened PMNs were not yet present in the circulating blood pool. Thus a pharmacological approach aimed at suppressing leucocyte activation before or during reperfusion therapy may be feasible.

Acute Disease↗

Controlled trial of inhaled budesonide in patients with cystic fibrosis and chronic bronchopulmonary Psuedomonas aeruginosa infection.

The efficacy and safety of anti-inflammatory treatment with inhaled glucocorticosteroids in patients with cystic fibrosis (CF) and complicating chronic Pseudomonas aeruginosa (P.a.) lung infection was studied in a placebo-controlled, parallel, double-blind single center trial. Active treatment consisted of budesonide dry powder, 800 microg twice daily, delivered from a Turbuhaler. The study period covered two successive 3-mo intervals between elective courses of intravenous anti-Pseudomonas antibiotics. Fifty-five patients entered the study, with a mean age of 20 yr and a mean FEV1 of 63% of predicted. Analysis of all patients entered, irrespective of trial adherence ("intention to treat"), showed a decrease in FEV1 in the first period of -0.032 L in patients on budesonide versus -0.187 L in patients on placebo (p = 0.08). The corresponding figures for the patients adhering to the protocol during the first period were -0.017 L versus -0.198 L (p < 0.05, confidence interval of the difference: -0.035 to +0.327 L). For all patients entered, as well as for patients adhering to the trial, there was always a trend in favor of budesonide, as judged by changes in FEV1 and FVC in both 3-mo periods. None of the patients had asthma, but the patients on budesonide had a mean improvement in histamine reactivity of +1.15 dose steps over the entire 6-mo period, as opposed to +0.017 dose steps in patients on placebo (p < 0.05). There was also a significant (p = 0.01) correlation between pre-trial histamine reactivity and the change in FEV1 in the first period in patients on budesonide. We conclude that inhaled glucocorticosteroids can be of short-term benefit in patients with CF and chronic P.a. infection and that those patients most likely to benefit from this treatment are patients with hyperreactive airways. Prolonged studies in larger number of patients are necessary to determine the long-term efficacy of this treatment.

Administration, Inhalation↗