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Biomedical subjects

J Verhoef

Publications and source records attributed to J Verhoef.

At least 325 records · Page 18Linked to original sources

Intranasal estradiol administration to oophorectomized women.

An open, dose-finding study into the effects of a new intranasal 17 beta-estradiol formulation using dimethyl-beta-cyclodextrin as a solubilizer was conducted in nine hysterectomized and oophorectomized patients with symptoms of estrogen deficiency. After nasal delivery of 0.34 mg estradiol in three patients, concentration-time curves of estradiol in serum and its metabolite estrone in plasma were established. High initial estradiol serum levels were found in the order of 5 nmol/l. Estrone/estradiol ratios were below unity, whereas, in contrast, estrone/estradiol ratios after oral administration of estradiol are known to exceed unity. Biological activity, as reflected by suppression of FSH and LH, was manifest. In nine patients symptoms of estrogen deficiency were assessed by a questionnaire. After 3 and 6 months of treatment, a clinically and statistically significant decrease (P less than 0.01) of the total score according to this questionnaire was found compared to the beginning of the study. Thus nasal delivery of estradiol using this cyclodextrin formulation has potential for further clinical evaluation.

Administration, Intranasal↗

Control of epidemic methicillin-resistant Staphylococcus aureus in a Dutch university hospital.

Between 1986 and 1989 a single strain of a methicillin- and multiply-resistant Staphylococcus aureus caused three distinct outbreaks at Utrecht University Hospital, involving 11, 19 and 32 patients, respectively. In all three episodes, members of staff were screened for MRSA carriage, and 58 persons were found to have positive nose cultures. In each outbreak it became necessary to isolate colonized and infected patients on a separate isolation ward. Staff carriers were also treated. Over the 18 months since the last outbreak, no new acquisitions of this epidemic MRSA strain have occurred. Between 1986 and 1989, the strain which caused the three outbreaks was not the only MRSA strain which was introduced into the hospital. Six other strains, which differed from the epidemic strain as shown by phage typing and antimicrobial susceptibility pattern, were found in single patients. The experience at Utrecht University Hospital illustrates the need for strict measures to eradicate epidemic strains of MRSA as well as the differences in "epidemicity" among various strains of MRSA.

Bacteriophage Typing↗

C-reactive protein in the diagnosis and management of infections in granulocytopenic and non-granulocytopenic patients.

The serum levels of C-reactive protein (CRP) were assayed in 64 non-granulocytopenic and 35 granulocytopenic patients with or without fever and infection. Most patients showed a direct CRP response within 24 hours after onset of fever (95% of non-granulocytopenic patients, 100% of granulocytopenic patients). The mean peak level of CRP in febrile patients with septicemia was 207 mg/l (median 214 mg/l) in non-granulocytopenic patients and 173 mg/l (median 168 mg/l) in granulocytopenic patients, and differed significantly (p less than 0.001) from that in febrile patients without positive blood cultures. A significant difference between patients with major and minor infections was also found (p less than 0.01). No significant difference in the CRP level was found between patients with microbiologically and clinically documented infections (p greater than 0.05), nor did the serum CRP levels differ between patients with infections due to gram-positive and gram-negative organisms. The most favorable cut-off limit for detection of an inflammatory process in this study was 25 mg/l. There was no quantitative difference between CRP levels measured by a latex-agglutination method and the nephelometry assay.

Adolescent↗

Analysis of Escherichia coli isolates from subjects with travellers' diarrhoea using DNA probes and serotyping.

Escherichia coli isolated from faeces of 54 healthy volunteers who visited Tunisia for eight days were examined. These volunteers participated in a randomized double-blind placebo-controlled study to establish whether ciprofloxacin could prevent travellers' diarrhoea. Escherichia coli strains isolated before travel, during episodes of travellers' diarrhoea, immediately after return and five weeks after return were serotyped and tested for the presence of virulence genes indicating diarrheogenic properties by hybridization with a set of four non-radioactively labelled DNA probes. Subjects receiving ciprofloxacin prophylactically to prevent travellers' diarrhoea were asymptomatic and no Escherichia coli could be cultured shortly after return home. Sixty-four percent of subjects (18/28) who did not receive antibiotic prophylaxis suffered from travellers' diarrhoea. Hybridization tests detected 8 enterotoxigenic Escherichia coli strains producing heat stable toxin, 13 enterotoxigenic strains producing heat labile toxin and 10 strains which produced both heat labile and stable toxin. Of the 31 probe positive strains, 29 (94%) were cultured from 11 volunteers with travellers' diarrhoea. A bacterial cause was thus determined in 61% of the volunteers who experienced travellers' diarrhoea.

Adult↗

Evaluation of the magnetic immuno PCR assay for rapid detection of Salmonella.

A new technique, the Magnetic Immuno PCR Assay (MIPA), has been developed for the detection of Salmonella. The assay utilizes magnetic particles coated with monoclonal antibodies against Salmonella to extract these bacteria from the sample. Trapped bacteria are lysed, and the supernatant, which contains bacterial DNA, is then subjected to the polymerase chain reaction (PCR) using primers from the Salmonella typhimurium origin of DNA replication to amplify a 163 bp region. The specificity of the primer set was tested in the PCR; amplification occurred with all 25 Salmonella strains tested but not with 19 other species of Enterobacteriaceae tested. A sensitivity of 100 cfu Salmonella typhimurium was achieved for the MIPA by visualization of the amplified products by ethidiumbromide stained agarose gel electrophoresis. A ten-fold higher sensitivity was obtained by Southern blotting of the amplified products. The presence of 10(7) cfu Escherichia coli did not interfere with these detection levels. The MIPA thus specifically detected 100 cfu of Salmonella within 5 h and may be potentially useful for rapid detection of Salmonella in clinical specimens and food.

Base Sequence↗

Species differences in impairment and recovery of alveolar macrophage functions following single and repeated ozone exposures.

Effects of single (0.4 ppm for 3, 6, or 12 hr) and repeated (0.4 ppm, 12 hr/day for 3 or 7 days) in vivo ozone exposures on rat and mouse alveolar macrophage functions and cell number were investigated. Single ozone exposure of rats resulted in a small (approximately 15%) decrease in Fc-receptor-mediated phagocytosis and phorbol ester-induced superoxide production by the alveolar macrophages and was followed by recovery above control levels within 12 hr of exposure. Repeated exposures of rats for up to 7 days did not alter alveolar macrophage functions, with the exception of the effects of 3 days of exposure on superoxide production (71 +/- 9% as compared with the controls). In mice, significant changes in alveolar macrophage functions were not observed until 12 hr of exposure (at that timepoint phagocytosis was 74 +/- 2%). Repeated ozone exposures of mice did not cause a further decrease in phagocytosis (at Day 7, 74 +/- 14%). Both after 3 and 7 days of repeated ozone exposure of mice, superoxide production by the alveolar macrophages was inhibited approximately 50%. In rats and mice, repeated ozone exposures led to an increase in the number of alveolar macrophages. In mice, this increase appeared at a later time point (at Day 7 vs Day 3) and was less pronounced (at Day 7, 139 +/- 9% vs 179 +/- 17%) as compared with rats. In summary, our data show that rat and mouse alveolar macrophages have different susceptibilities to both single and repeated in vivo ozone exposures.

Administration, Inhalation↗

The influence of the adjuvant Quil A on the epitope specificity of meningococcal lipopolysaccharide anti-carbohydrate antibodies.

Rabbits were immunized with immunotype L3,7,9 phosphoethanolamine (PEA) group containing oligosaccharide-tetanus toxoid conjugates both with and without the addition of the adjuvant Quil A. The epitope specificity of the antibodies present in these antisera was analysed in an immunotype L2 and L3,7,9 specific inhibition ELISA using the homologous and heterologous lipopolysaccharide, oligosaccharide and partial dephosphorylated oligosaccharide as inhibitors. Two groups of antisera could be identified. In one group of antisera, at least two antibody populations are present, namely directed against the PEA group containing determinants on immunotype L3,7,9 lipopolysaccharide and against immunotype L2 specific epitopes in which no PEA group is present. In the second group of antisera, one but probably more antibody populations are detected with a similar specificity towards the conserved epitopes of both immunotypes. In general, immunization with the conjugates only resulted in the induction of antibodies against the PEA group containing epitopes on the L3,7,9 lipopolysaccharide (80%). Antibodies directed against the conserved epitopes of both immunotypes are mainly evoked with the conjugates in combination with the adjuvant Quil A (80%). Although these results suggest that the epitope specificity of the antibodies induced depends on the use of Quil A, the influence of genetic factors cannot be excluded. At the moment it is not known whether the differences in epitope specificities are reflected in biological function of these antibodies. However, the induction of antibodies with clearly different epitope specificities after immunization of different rabbits with the same antigen stresses the importance of this kind of analysis when developing a vaccine based on oligosaccharide-protein conjugates.

Animals↗

Effects of ozone on cellular ATP levels in rat and mouse alveolar macrophages.

Hydrogen peroxide (H2O2) is thought to be a major intermediate in the toxicity of ozone. In a previous study we demonstrated that ATP depletion may play an important role in the H2O2-induced inhibition of the phagocytic functions of alveolar macrophages. Ozone exposure can adversely affect the phagocytic capacities of alveolar macrophages. In the present study we investigated whether a decrease in the cellular ATP concentration may underly the effects of ozone on alveolar macrophages. Neither following single (6 and 12 h) exposure nor repeated (12 h/day for 3 and 7 days) exposures of mice or rats to 0.4 ppm ozone, were decreased levels of ATP found in the alveolar macrophages. In contrast, repeated exposures of mice for 7 days to ozone led to a significant increase (1.5-fold) in the ATP content of the alveolar macrophages. In vitro ozone exposure of rat and mouse alveolar macrophages also did not lead to a decrease in the cellular ATP concentration. These results showed that ATP depletion does not play a role in the toxicity mechanism of ozone for alveolar macrophages.

Adenosine Triphosphate↗

Effect of sub-minimal inhibitory concentrations of ciprofloxacin and fleroxacin on the bacterial capsular antigen and opsonophagocytosis by human polymorphonuclear leukocytes.

We have studied the effect of sub-minimal inhibitory concentrations (sub-MIC) of ciprofloxacin and fleroxacin on capsulated (K+) and non-capsulated (K-) Gram-negative bacilli (Escherichia coli O1:K1, O7:K1, O1:K-, O7:K-, and Klebsiella oxytoca) as well as on Staphylococcus aureus and we investigated the interaction of antibiotic pretreated bacteria with human serum and polymorphonuclear leukocytes (PMN). Following overnight growth in the presence of 1/2 MIC of the antibiotics, bacteria were opsonized in human serum and incubated with PMN. Opsonophagocytosis was quantified as the ratio of uptake by PMN of radioactively labeled bacteria. Ciprofloxacin and fleroxacin enhanced the phagocytosis rate of E. coli K+ strains (control 5-10%; 1/2 MIC of ciprofloxacin and fleroxacin 70-80%) of K. oxytoca (control O-2%; 1/2 MIC of ciprofloxacin and fleroxacin 35-40%) as well as of Staph. aureus (control 5-10%; 1/2 MIC of the antibiotics 35-40%). Opsonophagocytosis of K- strains was not altered. The enhancement of opsonophagocytosis was a complement dependent process. Exposure of capsulated E. coli to ciprofloxacin as well as to fleroxacin resulted in decreased amounts of the capsular antigen.

Agglutination Tests↗

Immunization of mice with antibiotic-treated Escherichia coli results in enhanced protection against challenge with homologous and heterologous bacteria.

The murine immune response to Escherichia coli exposed to subminimal inhibitory concentrations of four antibiotics was investigated. Groups of mice were injected for 8 weeks with formalin-killed bacteria and subsequently challenged with 10 x LD50 of viable E. coli. Mice receiving saline only (controls) died within 24 h. The mortality of mice immunized with ciprofloxacin-treated E. coli was significantly lower than that of mice immunized with E. coli untreated or treated with other antibiotics. Sera from mice immunized with ciprofloxacin-treated bacteria showed better bacteriostatic capacity and enhanced production of antibodies that bound to homologous and heterologous lipopolysaccharide isolated from several smooth and rough gram-negative strains. The better protection observed in mice immunized with ciprofloxacin-treated E. coli was probably due to an enhanced production of antibodies to epitopes on lipopolysaccharide that became better exposed and so more accessible after treatment with ciprofloxacin.

Animals↗

A randomized study to compare oral fluconazole to amphotericin B in the prevention of fungal infections in patients with acute leukaemia.

In a prospective randomized study the efficacy of fluconazole (50 mg in one single daily dose) was compared with oral amphotericin B in suspension and tablets (each 200 mg four times daily) for prevention of colonization and subsequent infection by yeasts in 50 patients undergoing remission induction treatment for acute leukaemia. All patients received ciprofloxacin for prevention of bacterial infections. Fluconazole was as effective as amphotericin B in preventing severe local and disseminated fungal disease (one documented and one highly suspected infection in each group of patients). Fluconazole effectively prevented yeast colonization of the oropharynx but was less effective than amphotericin B in preventing colonization of the lower alimentary tract. Fifty-two percent of patients receiving fluconazole had persistent positive stool cultures as compared to 4% in the amphotericin B group (P less than 0.01). Fluconazole was better tolerated than amphotericin B. One patient developed an extended rash leading to the termination of fluconazole.

Acute Disease↗

Influence of polymorphonuclear granulocytes on the zona-free hamster oocyte assay.

To gain more insight into a possible detrimental influence of polymorphonuclear granulocytes (PMN) on fertility, PMN were added to spermatozoa and oocytes in the hamster oocyte assay. Addition of 2.5 x 10(6)/ml PMN to the sperm suspension at the start of the first incubation period (capacitation period) resulted in a significant decrease (P less than 0.03) in the number of decondensed sperm-heads per hamster oocyte (the decondensation index). When a concentration of 0.5 x 10(6)/ml PMN was used, no decrease was found. Stimulation of PMN by phorbol myristate acetate (PMA) did not enhance the inhibitory action of PMN. After incubation of oocytes with PMN for 1 h, an almost total inhibition of decondensation was observed. A significant inhibition could also be obtained when unstimulated PMN (2.5 x 10(6] were added at the start often 3 h coincubation period of spermatozoa and oocytes (P less than 0.004). The inhibition by PMN decreased to a nonsignificant level when the hydroxyl radical scavenger thiourea was present. The possible mechanisms by which PMN affect the results of the hamster oocyte assay and the possible implications of the presence of PMN in the genital tract are discussed.

Adult↗

Inhibition by lipid A-specific monoclonal antibodies of priming of human polymorphonuclear leucocytes by endotoxin.

Exposure to lipopolysaccharide (LPS) primes polymorphonuclear leucocytes (PMNL) for enhanced release of oxygen metabolites after subsequent stimulation. The metabolic response of human PMNL primed with LPS and stimulated with formyl-methionyl-leucyl-phenylalanine (FMLP) was measured by chemiluminescence (CL) as a parameter for endotoxic activity. Polymyxin B (PMB) and monoclonal antibodies (MAbs) with specificity for lipid A were tested for inhibition of the priming effect of Re LPS of Salmonella minnesota R595, Rc LPS of Escherichia coli J5 and smooth LPS of E. coli O111. The CL response of PMNL primed with Re LPS or Rc LPS was higher than that of PMNL primed with smooth LPS. Pre-incubation of rough or smooth LPS with PMB caused dose-dependent inhibition of priming of PMNL. Two IgM MAbs, 8-2 and 26-20, which recognise different epitopes on the hydrophobic part of lipid A, also completely prevented priming of PMNL by either rough or smooth LPS. The dose-dependent inhibitory effect of both MAbs was similar to the inhibition by PMB. These results indicate that the binding of MAbs to the hydrophobic part of lipid A is important in blocking lipid A-mediated effects.

Antibodies, Monoclonal↗

Characterisation and functional aspects of monoclonal antibodies specific for surface proteins of coagulase-negative staphylococci.

Mouse monoclonal antibodies (MAbs) raised against whole cells of Staphylococcus epidermidis strain 354 were characterised morphologically and functionally. Nine MAbs showed strong reactivity with coagulase-negative staphylococci (CNS). Only two MAbs were specific for CNS; both belonged to the IgG1 subclass, and one, MAb 36.4, reacted only with the strain used for immunisation. In immunoblotting, both CNS-specific MAbs 36.3 and 36.4 reacted strongly with cell-wall protein bands of 220 Kda of S.epidermidis strain 354 and weak reactivity was observed with a 110-Kda band. MAb 36.3 reacted also with 220-230 Kda bands of two other S.epidermidis strains (291 and ATCC 35984) and a 160-180 Kda band of S.epidermidis strain 354. Only MAb 36.4 promoted phagocytosis of strain 354 by polymorphonuclear leucocytes (PMNL) and monocytes, whereas MAb 36.3 and the other MAbs lacked this activity. Opsonisation of S. epidermidis with MAb 36.4 in the presence of complement enhanced uptake by PMNL, but not by monocytes. Furthermore, S.epidermidis strain 354 opsonised with MAb 36.4 induced chemiluminescence of PMNL. Immuno-gold electronmicroscopy with both MAbs 36.3 and 36.4 demonstrated a homogeneous distribution of gold particles on the surface as well as close to the surface of S.epidermidis.

Animals↗

Comparative in vitro antibacterial activity of sparfloxacin (AT-4140; RP 64206), a new quinolone.

The in vitro activity of sparfloxacin (AT-4140; RP 64206), a new fluoroquinolone, was compared with those of 10 other agents against 1,222 clinical isolates. Sparfloxacin and ciprofloxacin were the most active quinolones against members of the family Enterobacteriaceae and nonfermenting gram-negative bacilli; sparfloxacin had superior activity against gram-positive cocci in comparison with the activities of ciprofloxacin and the other quinolones tested (norfloxacin, lomefloxacin, and pefloxacin). Among the inhibited strains, several were resistant to the tested beta-lactam antibiotics or to aminoglycosides. The activity of sparfloxacin was not influenced by the medium that was used; lowering of the pH to 5 had a marked effect on the MICs for two strains each of Enterobacter cloacae and Pseudomonas aeruginosa and one strain each of Escherichia coli and Staphylococcus aureus; the MBC of sparfloxacin was within 1 to 2 dilution steps of the MIC for the strains that were tested.

4-Quinolones↗