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Biomedical subjects

M During

Publications and source records attributed to M During.

18 recordsLinked to original sources
PubMed2005

Detection of infectious Cryptosporidium oocysts by cell culture immunofluorescence assay: applicability to environmental samples.

In the past few years many waterborne outbreaks related to Cryptosporidium have been described. Current methods for detection of Cryptosporidium in water for the most part rely on viability assays which are not informative concerning the infectivity of oocysts. However, for estimation of the risk of infection with Cryptosporidium this information is required. For environmental samples the oocyst counts are often low, and the oocysts have been exposed to unfavorable conditions. Therefore, determination of the infectivity of environmental oocysts requires an assay with a high level of sensitivity. We evaluated the applicability of in vitro cell culture immunofluorescence assays with HCT-8 and Caco-2 cells for determination of oocyst infectivity in naturally contaminated water samples. Cell culture assays were compared with other viability and infectivity assays. Experiments with Cryptosporidium oocysts from different sources revealed that there was considerable variability in infectivity, which was illustrated by variable 50% infective doses, which ranged from 40 to 614 oocysts, and the results indicated that not only relatively large numbers of fresh oocysts but also aged oocysts produced infection in cell cultures. Fifteen Dutch surface water samples were tested, and the cell culture immunofluorescence assays were not capable of determining the infectivity for the low numbers of naturally occurring Cryptosporidium oocysts present in the samples. A comparison with other viability assays, such as the vital dye exclusion assay, demonstrated that surrogate methods overestimate the number of infectious oocysts and therefore the risk of infection with Cryptosporidium. For accurate risk assessment, further improvement of the method for detection of Cryptosporidium in water is needed.

Animals↗
PubMed2002

Replication competent helper functions for recombinant AAV vector generation.

Adeno-associated virus (AAV) is a promising gene transfer vector tested in both animal studies and human clinical trials. However, current production methods are generally inefficient and require improvements to meet the increasing clinical need for economical, high titer and high quality rAAV vectors. The inefficiency of the current systems largely arises from the AAV helper function, which contains only the AAV coding region but lacks inverted terminal repeats. The terminal repeats were originally removed to prevent replication competent AAV contamination. Here we designed a novel and highly efficient rAAV helper function containing AAV terminal repeats. The new helper function not only mimics the wild-type AAV growth as it replicates along with the vector plasmid, but also restores the cis regulating function of the AAV terminal repeats. Addition of heterologous introns to the helper genome and use of a mutant AAV terminal repeat defective in packaging effectively controls the contamination of replication competent AAV particles. This new strategy also performs better in AAV producing cell lines than those based on non-replicating AAV rep and cap genome.

Cell Line↗
PubMed2000

c-Jun promotes neurite outgrowth and survival in PC12 cells.

We investigated the function of c-Jun in PC12 cells by transfecting them with a plasmid containing a c-Jun cDNA transcription cassette. Transfected cells expressed high levels of c-Jun mRNA and protein and demonstrated an increase in both AP-1 DNA binding and gene activation. The c-Jun over-expressing cells showed marked neurite outgrowth but no evidence of spontaneous cell death. In fact, c-Jun over-expressing cells were more resistant to okadaic acid-induced apoptosis. The process outgrowth was not indicative of a full neuronal differentiation response as the transfected PC12 cells did not display action potentials when examined with whole-cell patch-clamping. The phosphorylation of c-Jun on serine 73 appears to be important for this neurite sprouting effect as mutagenesis at this site reduced sprouting whereas a serine 63 mutant tended to increase sprouting. Thus, in PC12 cells c-Jun expression does not induce apoptosis, but rather functions as a neurite outgrowth and neuronal survival signal.

Action Potentials↗
PubMed1997

The effect of leptin is enhanced by microinjection into the ventromedial hypothalamus.

To determine whether changes in food intake produced by leptin involve targeting the hormone to distinct central nervous system regions, guide cannulas were positioned stereotaxically into three brain regions--the ventromedial hypothalamus (VMH) (bilaterally, n = 6), the dorsal raphe nucleus (n = 3), and the lateral ventricle (n = 3)--of nonobese male rats (400-500 g). Daily food intake and body weight changes were measured during twice-daily injections of saline (0.1 microl) followed by recombinant human leptin (0.05 microg) for 3 days via the brain cannulas. VMH-injected rats also were followed during a postleptin saline recovery interval. This small dose of leptin did not change food intake or body weight from that during the preceding saline injection period in ventricle-injected or dorsal raphe-injected rats. In sharp contrast, VMH-injected rats ate much less food (56 +/- 8% basal) and lost 9 +/- 3 g/day or 5% of their body weight during 3 days of leptin administration. VMH-injected animals fully recovered from leptin-induced effects within 3 days. We conclude that small doses of leptin that do not effect eating behavior when delivered to the ventricle or the dorsal raphe (another brain region believed to regulate feeding), suppress food intake when injected into the VMH. These data suggest that the VMH or a brain region in close proximity to it is a key target for the biological actions of leptin.

Animals↗
PubMed1994

Community-based study of the incidence of gastrointestinal diseases in The Netherlands.

The incidence of gastrointestinal diseases was studied in a community-based study in four regions of The Netherlands. Two grades of severity were distinguished--1: diarrhoea or vomiting and at least 2 additional symptoms within the period of 1 week, and 2: diarrhoea or vomiting and at least 2 additional symptoms occurring on the same day lasting at least 2 days within the period of 1 week. The incidence of gastrointestinal episodes was calculated to be 630 for grade 1 and 180 for grade 2 disease per 1000 person-years, after correction for age and sex. The incidence was higher for women than for men (relative risk 1.25) and lower for those in the 19-64-year-old age group when compared to those younger or older (relative risk 0.75 and 0.40, respectively). Independent of the degree of severity of the symptoms, about 20% of the patients had consulted a general practitioner, about half in person and half by telephone. It is concluded that community studies are essential to assess the real incidence of gastrointestinal diseases in the population.

Adolescent↗
PubMed1993

[123I]iomazenil SPECT imaging demonstrates significant benzodiazepine receptor reserve in human and nonhuman primate brain.

SPECT imaging with [123I]iomazenil was used to measure benzodiazepine (BZ) neuroreceptor occupancy of the agonist lorazepam administered at therapeutically relevant doses in humans and supratherapeutic doses in monkeys. Lorazepam at therapeutic doses (0.03 mg/kg, i.v.) administered 90 min after the bolus injection of [123I]iomazenil had no statistically significant effect (P > 0.12) on the washout rates of regional brain activities compared to that in control subjects, although human subjects demonstrated marked sedation from the lorazepam. In baboons, the effects of higher doses of lorazepam (cumulative 0.5 mg/kg) were examined in a stepwise displacement paradigm. The in vivo potency was expressed as the ED50 (or dose required to displace 50% of receptor bound activity) and was equal to 0.34 +/- 0.01 mg/kg (mean +/- SD, n = 12). Log-logit analyses of displacement data corrected for endogenous washout showed that therapeutic doses of lorazepam were associated with < 3% BZ receptor occupancy. To examine if endogenous GABA modulates potency of the BZ agonist, the ED50 of lorazepam was compared with and without concurrent administration of tiagabine, a GABA reuptake inhibitor. These experiments were designed to measure an in vivo GABA shift of agonist potency. In vivo microdialysis demonstrated that tiagabine (up to 1 mg/kg, i.v.) increased extracellular GABA levels up to 200% of baseline, but these doses had only a minimal enhancement of lorazepam's potency to displace [123I]iomazenil. This study strongly suggests that single therapeutically relevant doses of lorazepam occupy a relatively small percentage (i.e. < 3%) of BZ receptors and that BZ binding sites have a significant (i.e. > 97%) receptor reserve.

Adult↗
PubMed1990

[Aeromonas species as cause of diarrhea and infections outside the gastrointestinal tract in The Netherlands].

In a national survey in the period May 1986-December 1987, Aeromonas was isolated from 277 out of 16,857 (1.6%) samples of watery, bloody or mucous stool, from patients with diarrhoea. There was a clear seasonal pattern (less than 1% in winter, up to 3% in summer). A. caviae was isolated most frequently (49%), followed by A. sobria (35%) and A. hydrophila (15%). Some non-identifiable strains were isolated as well. Aeromonas were isolated in particular from faeces of patients aged over 70 years (predominantly A. sobria) or under 5 years (predominantly A. caviae). In 67% Aeromonas was isolated as the only possible bacterial cause of diarrhoea, but in 33% other enteropathogenic bacteria were found as well (17% Campylobacter, 14% Salmonella, 2% Shigella). In addition, all Aeromonas isolates were collected which were obtained in normal diagnostic activities in the participating laboratories, among others from blood, from pus or wound fluid, from faecal samples which did not meet the above mentioned criteria. A. caviae was the dominant species in 'other' faeces and 'various' body sites but was not isolated from blood. The results of this study do not indicate that routine examinations for Aeromonas in faeces of patients with diarrhoea are necessary.

Adolescent↗
PubMed1990

The presence of Aeromonas in drinking water supplies in The Netherlands.

The occurrence of Aeromonas in raw, treated and distributed waters of 20 different treatment stations in the Netherlands was studied over a one-and-a-half year period. River water yielded highest numbers (greater than 10,000 cfu/100 ml) of predominantly anaerogenic strains. In open storage reservoirs for river water or dune infiltrate, numbers were usually between 1,000 and 10,000/100 ml with a majority of aerogenic strains, in particular A. sobria. River water after dune infiltration or pure dune water, collected in closed systems, as well as aerobic or anaerobic groundwater, were usually free of Aeromonas in 100 ml. Treated waters showed low counts (usually less than 10 cfu/100 ml), irrespective of raw water source. Regrowth of aeromonads occurred in 16 out of 20 distribution systems examined. Geometric means counts (2nd half of 1986) in these systems varied between 1 and 440 cfu/100 ml and maximum counts between 10 and 3300 cfu/100 ml. Aeromonas-densities were related to temperature and residence time, but not to total organic matter or heterotrophic plate counts. Regrowth occurred particularly in drinking water derived from anaerobic groundwater containing methane. A. hydrophila was the most frequently isolated species from distribution waters, but A. caviae and A. sobria were predominant in a few systems.

Aeromonas↗
PubMed1989

Effects of running the Boston marathon on plasma concentrations of large neutral amino acids.

Plasma large neutral amino acid concentrations were measured in thirty-seven subjects before and after completing the Boston Marathon. Concentrations of tyrosine, phenylalanine, and methionine increased, as did their "plasma ratios" (i.e., the ratio of each amino acid's concentration to the summed plasma concentrations of the other large neutral amino acids which compete with it for brain uptake). No changes were noted in the plasma concentrations of tryptophan, leucine, isoleucine, nor valine; however, the "plasma ratios" of acid patterns may influence neurotransmitter synthesis.

Amino Acids↗
PubMed1989

Reducing sleep disruptions in young children: evaluation of therapist-guided and written information approaches: a brief report.

Regular night waking of young children is one of the most common problems encountered by parents. This study compared a standardized night waking programme that involved organized bedtime routines, procedures for settling the child and for the handling of crying, calling out and getting out of bed, with a group which received written information only and a waiting list control group. The children receiving the standard programme and those receiving written information only showed significant improvement over children in the waiting list group. This result supports the use of written parent instructions with or without therapist support.

Behavior Therapy↗
PubMed1988

Evaluation of the Anderson Baird-Parker direct plating method for enumerating Escherichia coli in water.

The direct plating (DP) method for enumerating Escherichia coli in food was adapted for water analysis by membrane filtration and a standardized protocol was described. The DP method was found to give equal or better recoveries of E. coli than a membrane filtration method using 0.1% sodium lauryl sulphate agar; the repeatability of the DP method was markedly better. The necessity to transfer membranes from the non-selective medium tryptone soy agar (TSA) to the selective medium tryptone bile agar (TBA) after pre-incubation for 4 h was considered disadvantageous for practical purposes. A double-layer method, where the membrane filter is placed on a layer of TSA poured over TBA, with incubation in an incubator that automatically switches from 37 degrees to 44 degrees C after 4 h, was found to be an acceptable alternative. Recovery of E. coli and inhibition of competitive flora were equal or only slightly less than for the standard DP method.

Bacteriological Techniques↗
PubMed1987

Ampicillin-dextrin agar medium for the enumeration of Aeromonas species in water by membrane filtration.

Published selective media were evaluated for the isolation of Aeromonas spp. from environmental samples by membrane filtration. Satisfactory recoveries were obtained only with mA agar (Rippey & Cabelli) and dextrin-fuchsin-sulphite agar (Schubert), but neither was sufficiently selective. The positive aspects of these two media were combined in a new medium, ampicillin-dextrin agar. Recovery from pure cultures and environmental samples was optimal at an ampicillin concentration of 10 mg/l and incubation for 24 h at 30 degrees C under aerobic conditions, and specificity was high (i.e. confirmation rate usually greater than 90%, no false negative colonies encountered). The medium can also be used for isolation of Aeromonas spp. from sea water provided that the vibriostatic agent 0/129 is added at 50 mg/l.

Aeromonas↗
PubMed1986

C-390 as sole selective agent for isolation of Pseudomonas aeruginosa from hospital waste water.

Pseudomonas aeruginosa was recovered (in numbers ranging from 10(2) to 10(5) colony-forming units per millilitre) from heavily contaminated hospital waste water when grown at 41.5 degrees C on a differential medium agar containing 9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan (C-390) at a final concentration of 30 micrograms/mL. The medium appeared to be highly selective for P. aeruginosa with 95-100% of all colonies isolated from four different hospital waste waters being identified as P. aeruginosa. Many strains of P. aeruginosa isolated from hospital waste waters failed to hydrolyse casein when grown on skim milk agar and this medium appeared to restrict pigment production to only pyoverdin (detectable only under ultraviolet light). However, most strains were capable of casein hydrolysis when grown on a modified skim milk medium.

Acridines↗
PubMed1985

Model studies on a membrane filtration method for the enumeration of coagulase-positive staphylococci in swimming-pool water using rabbit plasma-bovine fibrinogen agar.

The recovery of Staphylococcus aureus from swimming-pool water by membrane filtration was studied in model experiments. On the nonselective medium tryptone soya agar (TSA) there was no difference in counts of noninjured S. aureus with all membrane filters tested and with pour plates. Chlorine-injured S. aureus was enumerated most efficiently on TSA by Gelman Tuffryn HT-450 and Sartorius SM 13806 filters. Tuffryn filters were also most productive when used in combination with the selective medium rabbit plasma - bovine fibrinogen agar (RPFA). Other filters, particularly Gelman GN-6 and Millipore HAWP, when used on RPFA were shown to have a synergistic inhibitory effect on both noninjured and chlorine-injured S. aureus. This effect was not found on Baird-Parker agar. Using Tuffryn filters, counts on RPFA were equal to those on TSA for noninjured S. aureus and 0.1-2.0 log units less for chlorine-injured S. aureus. Despite this, the possibility for reading the in situ coagulase reaction for individual colonies on RPFA is considered such an advantage of this medium that its general use for enumeration of S. aureus in swimming pools is recommended. Further studies should be carried out to allow better resuscitation.

Animals↗
PubMed1985

Comparative study of membrane filtration and enrichment media for the isolation and enumeration of Pseudomonas aeruginosa from sewage, surface water, and swimming pools.

The recovery of Pseudomonas aeruginosa on several selective culture media was tested using raw sewage and secondary sewage effluent samples as well as spiked chlorinated imitation swimming water and samples from whirlpools. mPA-medium B gave good recovery of both vital and chlorine-injured P. aeruginosa and selectivity was greater than 90% when analysing whirlpool samples. It is therefore the medium recommended for examination of chlorinated swimming pools. When analysing sewage polluted water with the mPA-B medium, reduced selectivity was noted from low verification rates and from overgrowth by competitive flora. A modified medium (mPA-D; addition of cetrimide, omission of sulphapyridine and actidione) was more selective and sufficiently recovered noninjured cells. Chlorine-injured cells were completely inhibited, however. C-390 (9-chloro-9-(4-diethylaminophenyl)-10-phenylacridan) was confirmed to be highly selective for P. aeruginosa when used in spread plates at a concentration of 30 micrograms/mL; P. aeruginosa was slightly inhibited. However, the medium could not be used with conventional membrane filtration techniques, because cellulose ester filters interfered with the selective action of C-390. Selectivity could be improved by using Gelman Tuffryn (polysulphone) filters and increasing the C-390 concentration to 120 micrograms/mL. At this concentration, however, the medium was strongly inhibitory to P. aeruginosa; resuscitation only partially improved recovery. Two other membrane filtration media were tested. Both cetrimide - nalidixic acid agar and Drake's medium No. 19 were inhibitory to chlorine-injured cells. Several types of membrane filters were tested and there was little difference between them.(ABSTRACT TRUNCATED AT 250 WORDS)

Cetrimonium Compounds↗