Women's motives for not participating in preconception counseling: qualitative study.

AIMS: Information about risk factors and preventive measures given before conception is estimated to prevent 15-35% of adverse pregnancy outcomes. We aimed to identify women's motives for not responding to an invitation for preconception counseling (PCC) from their general practitioner. METHODS: A purposive sample of 11 women who did not respond to an invitation for PCC and who became pregnant within 1 year was interviewed. RESULTS: Three key themes influencing nonresponse emerged from the data: perceived knowledge, perceived lack of risk and a misunderstanding of the aim of PCC. CONCLUSION: For successful future implementation of PCC, a more tailored approach may be necessary for certain (groups of) women, addressing the reasons why women do not consider themselves part of the target group for PCC.

The development and validation of a handicap questionnaire for children with a chronic illness.

INTRODUCTION: This paper describes the development and initial psychometric evaluation of the Handicap Scale for Children (HSC). This questionnaire is based on the London Handicap Scale (LHS), a valid and reliable utility instrument for measuring social participation in adults. METHODS: A multidisciplinary research group was involved in developing the HSC. The questionnaire was tested in 114 children with a chronic disease and 239 healthy children in the 8-18 age range. Relating the Health Utility Index Mark 3 (HUI3) attributes to corresponding HSC scores tested the assumption that a negative health status would lead to participation problems. RESULTS: Questionnaire development resulted in a five-dimension questionnaire: mobility, physical independence, daily activities, social integration and orientation. Each dimension included one item with a six-point response scale. A higher score indicates greater handicap. Feasibility testing with 10 children showed that none of the children experienced difficulties in filling in the questionnaire. Conceptual validity, measured by correlations between the dimensions of the HSC and HUI3, was satisfactory. As expected, moderate correlation coefficients between predefined pairs of HUI and HSC attributes were found; other correlation coefficients were low. Criterion validity was also satisfactory, as shown by large differences between the healthy and the chronically ill group and by several criteria within the chronically ill group. CONCLUSION: Based on this initial evaluation, the questionnaire seems feasible and valid for use with children in the age range 8-18 years.

Colocalization of androgen, estrogen and cholinergic receptors on cultured astrocytes of rat central nervous system.

By means of immunohistochemical and electrophysiological methods, we have investigated the presence of androgen receptors on astrocytes in explant and primary cultures from various regions of rat central nervous system. Our studies have shown that a great number of astrocytes and neurones express androgen receptors as recognized by a specific monoclonal antibody. Immunoreactivity was mainly distributed over the soma of the astrocytes, the nuclei being intensely stained. In contrast, glial processes were only faintly stained or not stained. Double-immunostaining studies have provided evidence for a colocalization of androgen and estrogen alpha- and beta-receptors on many astrocytes. Furthermore, there was also a coexistence of glial androgen receptors with cholinergic muscarinic and nicotinic sites. Our immunohistochemical findings are supported by electrophysiological investigations demonstrating that 5alpha-androstan, 17beta-estradiol as well as the cholinergic agonists muscarine and nicotine caused hyperpolarizations on the same astrocytes. Our studies suggest that there is a coexistence of functional receptors for androgen, estrogen as well as for the cholinergic agonists on glial cells. Further investigations are needed to elucidate the physiological role of glial androgen, estrogen and cholinergic receptors and to define their function in neurodegenerative diseases.

Colocalization of neurotransmitter receptors on astrocytes in explant cultures of rat CNS.

In recent years evidence has accumulated that astrocytes express functional receptors for a variety of neurotransmitters/neuromodulators. By means of electrophysiological and combined autoradiographic and immunohistochemical methods we have demonstrated the colocalization of cholinergic, adrenergic and peptidergic receptors on astrocytes in explant cultures from various regions of rat central nervous system. A great number of biochemical and electrophysiological studies from other laboratories have shown that most of the neurotransmitters exert their effects on second messenger systems and on Ca2+-activated K+-channels. Furthermore, certain neurotransmitters are involved in the regulation of energy metabolism by stimulating enzymatic breakdown of glycogen in astrocytes. It was suggested that there is a cross-talk between the various neurotransmitter receptors on the glial membrane and that these receptors act in a synergistic or antagonistic way. The coexistence of cholinergic and peptidergic receptors on astrocytes is of great interest since both neurotransmitter systems are involved in cognitive functions and are impaired in patients with Alzheimer's dementia. The question is therefore raised whether not only neurones but also astrocytes might be involved in neurodegenerative disorders such as Alzheimer's disease.

Histochemical and electrophysiological evidence for estrogen receptors on cultured astrocytes: colocalization with cholinergic receptors.

By means of autoradiographic and immunohistochemical methods it was demonstrated that astrocytes in explant and primary cultures of rat neocortex, hippocampus, preoptic area and spinal cord express estrogen alpha- and beta-receptors. Immunoreactivity was mainly distributed over the soma, the nuclei being more intensely stained. Combined autoradiographic and immunohistochemical studies as well as double-immunostaining revealed a colocalization of estrogen alpha- and beta-receptors on many astrocytes. There was also a coexistence of estrogen receptors and cholinergic muscarinic and nicotinic sites. Electrophysiological investigations have shown that 17beta-estradiol induced hyperpolarizations on the majority of astrocytes in explant cultures of hippocampus and spinal cord, providing evidence for the existence of functional estrogen receptors on these cells. Furthermore, on the same astrocytes, 17beta-estradiol, muscarine and nicotine caused hyperpolarizations, suggesting a coexistence of receptors for estrogen and the cholinergic agonists on glial cells. The presence of glial estrogen receptors and their colocalization with cholinergic receptors is discussed with respect to the effects of these neurotransmitters/neuromodulators in development and maturation of the central nervous system, as well as to neurodegenerative events such as Alzheimer's disease.

Cellular localization of estrogen receptors on neurones in various regions of cultured rat CNS: coexistence with cholinergic and galanin receptors.

Autoradiographic studies have shown that many neurones in explant cultures of rat neocortex, hippocampus, preoptic area and spinal cord express binding sites for [3H]-estradiol which are distributed over the cell bodies and primary processes. By means of immunohistochemistry, it was observed that neurones were labelled by monoclonal antibodies against estrogen alpha-receptors and a polyclonal antibody against estrogen beta-receptors. Immunoreactivity was distributed over the soma and primary processes of the cells, the nuclei being more intensely stained. Double-immunostaining revealed a colocalization of estrogen alpha- and beta-receptors on approximately half of the neurones in cultures from neocortex and hippocampus whereas in cultures from preoptic area and spinal cord only few cells were double-stained. On many neurones, a coexistence of estrogen receptors and cholinergic muscarinic or nicotinic sites was found. Furthermore, combined autoradiographic and immunohistochemical studies have shown a colocalization of receptors for estrogen and the neuropeptide [125I]-galanin. The coexistence of estrogen and cholinergic sites as well as of estrogen and galanin receptors on the same neurones are discussed with respect to neurodegenerative events such as Alzheimer's disease.

Evidence for the existence of galanin receptors on cultured astrocytes of rat CNS: colocalization with cholinergic receptors.

The cellular localization of binding sites for [125I]galanin was studied in explant cultures of rat neocortex, cerebellum, locus coeruleus and spinal cord by means by of autoradiography. Binding sites for the peptide were observed on a great number of astrocytes in all CNS regions studied. In addition to astrocytes, many neurones were intensely labelled by [125I]galanin. Binding of [125I]galanin (10(-8) M) to both astrocytes and neurones was markedly reduced or inhibited by the unlabelled peptide at high concentration (10(-6) M), suggesting 'specific' binding of the radioligand. Evidence for the colocalization of galanin and cholinergic receptors on astrocytes was provided by combined autoradiographic and immunohistochemical studies. Many astrocytes were labelled by [125I]galanin and immunostained with antibodies to either muscarinic or nicotinic receptors. Electrophysiological studies revealed that addition of galanin (10(-9) to 10(-7) M) to the bathing fluid caused a dose-dependent hyperpolarization of the majority of astrocytes studied. When galanin (10(-8) M) and the cholinergic agonists muscarine and nicotine (10(-6) M) were tested on the same astrocyte, all three compounds induced a hyperpolarization, suggesting a colocalization of functional galanin and cholinergic receptors on the glial membrane.

Autoradiographic studies on the uptake of 3H-dopamine by neurons and astrocytes in explant and primary cultures of rat CNS: effects of uptake inhibitors.

The cellular localization of the uptake of 3H-dopamine was studies in explant and primary cultures from various regions of rat central nervous system by means of autoradiography. In explant cultures of substantia nigra, 3H-dopamine was taken up by cell bodies and processes of many neurons. In cultures from striatum, cerebellum and spinal cord, neuronal cell bodies were not labelled, whereas outgrowing nerve fibres revealed intense uptake of the monoamine. Uptake of 3H-dopamine by neurons was Na(+)- and temperature-dependent, suggesting an active uptake mechanism. In explant cultures, astrocytes did not accumulate 3H-dopamine, whereas in primary cultures, which were prepared from the same regions of rat central nervous system as the explant cultures, astrocytes also revealed uptake of this monoamine. The intensity of labelling was dependent on the incubation time. Little uptake of 3H-dopamine was observed after an incubation time of 5 min and only after 10-15 min did the astrocytes show moderate labelling. Uptake of 3H-dopamine by astrocytes was not Na(+)- and temperature-dependent, indicating that glial cells do not possess an active uptake mechanism for this monoamine. This is consistent with biochemical investigations by other laboratories, demonstrating that astrocytes accumulate 3H-dopamine by a facilitated diffusion system. Addition of the uptake inhibitors nomifensine or GBR 12909 to explant cultures markedly reduced or inhibited uptake of 3H-dopamine by neurons at a concentration of 10(-6) M. In contrast, accumulation of 3H-dopamine by astrocytes in primary cultures was only slightly affected by nomifensine at 10(-6) M. At the highest concentration used (10(-5) M), nomifensine also blocked the uptake of 3H-dopamine by astrocytes. Our finding that GBR 12909 almost completely inhibited the uptake of 3H-dopamine by astrocytes already at 10(-6) M suggests that this compound is a more potent inhibitor of the glial uptake of dopamine than nomifensine.

Expression of GABA(A) receptors by reactive astrocytes in explant and primary cultures of rat CNS.

The presence of GABA(A)-receptors on astrocytes was studied in explant and primary cultures of rat cerebellum, hippocampus and spinal cord by means of immunohistochemistry. For these studies we have used the monoclonal antibody bd 17 against the beta2- and beta3-subunits of GABA(A)-receptor. In explant cultures many neurones were intensely stained with the GABA(A)-receptor antibody whereas adjacent astrocytes revealed little or no immunoreactivity. In the far outgrowth zone of explant culture, however, many immunostained astrocytes were observed. In primary astrocyte cultures, only a few cells were stained by the antibody. Astrocytes which became reactive after producing an artificial scar or after addition of certain compounds such as dibutyryl cyclic AMP, interleukin-6, basic fibroblast growth factor and kainic acid, also revealed GABA(A)-receptor immunoreactivity. Furthermore, these astrocytes were intensely stained for glial fibrillary acidic protein and vimentin. From our studies we conclude that only a sub-population of normal astrocytes are immunopositive for the GABA(A)-receptor antibody whereas astrocytes which become reactive following injury of the tissue or after addition of dibutyryl cyclic AMP, the cytokine interleukin-6, fibroblast growth factor or the neurotoxin kainic acid express GABA(A)-sites.

Action and binding of the endothelin antagonist bosentan on astrocytes of cultured rat central nervous system. Electrophysiological and autoradiographic studies.

Our autoradiographic studies demonstrate that astrocytes in explant cultures of rat central nervous system possess binding sites for the first orally active, mixed, nonpeptide endothelin receptor antagonist [3H] bosentan. Binding of [3H]bosentan was inhibited by unlabelled bosentan and endothelin-1 at high concentrations, suggesting specific binding of the antagonist. Electrophysiological studies have revealed that bosentan reversibly blocked the depolarizations by endothelin but not by angiotensin II, indicating that the antagonist specifically antagonizes the action of endothelin on the glial membrane. This is consistent with biochemical studies from other laboratories demonstrating that bosentan did not interfere with binding of angiotensin II. The availability of bosentan, a potent and selective endothelin receptor antagonist should help to elucidate the role of endothelin on astrocyte function.

Autoradiographic studies on the uptake of 3H-noradrenaline and 3H-serotonin by neurones and astrocytes in explant and primary cultures of rat CNS: effects of antidepressants.

Autoradiographic studies were made on the uptake of 3H-noradrenaline and 3H-serotonin in explant cultures and primary astrocyte cultures from various regions of rat central nervous system (cortex, cerebellum, locus coeruleus, nucleus raphé, spinal cord). In explant cultures from locus coeruleus and nucleus raphé cell bodies and processes of many neurones revealed intense labelling by 3H-noradrenaline and 3H-serotonin, respectively. In cultures from cortex, cerebellum and spinal cord the cell bodies of neurones did not show labelling by the monoamines but many nerve fibres in the outgrowth zone had taken up 3H-noradrenaline and 3H-serotonin. Astrocytes in explant cultures did not take up 3H-noradrenaline and 3H-serotonin whereas astrocytes in primary cultures showed heavy uptake of both monoamines. In contrast, amino acid transmitters such as 3H-GABA and 3H-glutamate were accumulated by astrocytes in explant as well as in primary cultures. Uptake of both 3H-noradrenaline and 3H-serotonin by neurones and astrocytes was considerably reduced or inhibited in Na(+)-free incubation medium or at low temperature, suggesting an active uptake mechanism. Addition of the antidepressants maprotiline and (+)oxaprotiline inhibited the uptake of 3H-noradrenaline by neuronal cell bodies and fibres in explant cultures and by astrocytes in primary cultures. The uptake of 3H-serotonin by neurones and astrocytes was blocked by citalopram and paroxetine. Our studies demonstrate that astrocytes in primary cultures are able to actively take up 3H-noradrenaline and 3H-serotonin whereas there was no uptake of monoamines into astrocytes in explant cultures, suggesting that there is a difference between astrocytes in different culture systems (explant cultures vs primary cultures) with respect to the uptake of monoamine transmitters.

Autoradiographic and electrophysiological evidence for the existence of neurotensin receptors on cultured astrocytes.

By means of autoradiography we have studied the cellular localization of binding sites for [3H]neurotensin and its nonpeptide receptor antagonist [3H]SR-48692 in explant cultures of rat neocortex, striatum, brain stem and spinal cord. Binding sites for the peptide and its antagonist were observed on a great number of astrocytes in all CNS regions studied. Simultaneous staining of the cultures with a monoclonal antibody against glial fibrillary acidic protein has shown that the labelled cells in the outgrowth zone of the cultures were glial fibrillary acidic protein-positive and could therefore be identified as astrocytes. In addition to astrocytes, many neurons and outgrowing nerve fibres were labelled by the radioligands. Binding of [3H]neurotensin and [3H]SR-48692 (10(-8)M) to neurons and glial cells was markedly reduced or inhibited by the unlabelled compounds at high concentration (10(-6)M), suggesting "specific" binding of the radioligands. Electrophysiological studies have shown that addition of neurotensin to the bathing solution caused a hyperpolarization of the majority of astrocytes tested. There was a dose-response relationship between the magnitude of the hyperpolarization and the concentration of the peptide (10(-10)-10(-7)M); 10(-10)M being the threshold concentration. The specificity of the action of neurotensin was confirmed by the selective nonpeptide neurotensin receptor antagonist SR-48692 which reversibly blocked or markedly reduced the hyperpolarization by the peptide on all astrocytes tested. Our electrophysiological findings together with our autoradiographic data provide strong evidence for the presence of specific and functional neurotensin receptors on astrocytes.

Coexistence of cholinergic and somatostatin receptors on astrocytes of rat CNS.

Electrophysiological studies have shown that somatostatin (SOM; 10(-8) and 10(-7) M) causes a hyperpolarization of the majority of astrocytes in explant cultures of rat spinal cord and cortex. When SOM and the cholinergic agonists muscarine and nicotine (10(-6) M) were tested on the same cell, all three compounds produced hyperpolarizations, suggesting a colocalization of functional cholinergic and SOM receptors on the glial membrane. Combined immunohistochemical and autoradiographic binding studies demonstrating that almost all astrocytes which were immunostained by the monoclonal muscarinic or nicotinic antibodies were also intensely labelled by 125I-SOM, provide further evidence for the coexistence of cholinergic and SOM receptors on astrocytes.

Binding of cholecystokinin, bombesin and muscarine to neurons and astrocytes in explant cultures of rat central nervous system: autoradiographic and immunohistochemical studies.

The cellular localization of binding sites for the gastrointestinal peptides [3H]cholecystokinin and [125I]bombesin as well as the cholecystokininB-antagonist [3H]L-365,260 was investigated in explant cultures of rat cortex, cerebellum, brainstem and spinal cord using autoradiographic techniques. Many neurons in cortical, brainstem and spinal cord cultures revealed intense labelling of the radioligands whereas cerebellar neurons showed only little binding. In addition to neurons, binding sites for these peptides were also observed on astrocytes. Labelling of glial cells in cerebellar cultures was usually weaker than in the other CNS areas studied, suggesting a certain specialization of astrocytes in various brain regions. By means of combined immunohistochemical and autoradiographic techniques it was demonstrated that many neurons and astrocytes which expressed binding sites for [3H]cholecystokinin, [3H]L-365,260 and [125I]bombesin were also immunostained by the monoclonal muscarinic receptor antibody M 35 providing evidence for a co-localization of peptidergic and cholinergic receptors on the membrane of these cells. Our autoradiographic findings suggesting the presence of receptors for cholecystokinin and bombesin on astrocytes are supported by electrophysiological studies demonstrating that both peptides induce a hyperpolarization of glial cells.

Colocalization of binding sites for somatostatin, muscarine and nicotine on cultured neurones of rat neocortex, cerebellum, brain stem and spinal cord: combined autoradiographic and immunohistochemical studies.

The cellular localization of binding sites for [125I]1-tyramine somatostatin ([125I]SS) was studied in explant cultures of rat CNS by autoradiography. In cultures from cortex, brain stem and spinal cord many neurones revealed binding sites for the peptide whereas in cerebellar cultures only little binding of [125I]SS was observed. In addition to neurones, astrocytes were also labelled by the peptide. By combined immunohistochemical and autoradiographic techniques, it was demonstrated that the majority of neurones which expressed binding sites for [125I]SS were also immunostained by the monoclonal cholinergic muscarinic or nicotinic receptor antibodies (M 35 and W 6, respectively), providing evidence for a colocalization of cholinergic and somatostatin receptors on the neuronal membrane.

Electrophysiological evidence for the presence of receptors for cholecystokinin and bombesin on cultured astrocytes of rat central nervous system.

The action of cholecystokinin (CCK) and bombesin (Bom) was studied on the membrane potential of astrocytes in explant cultures of rat cortex, cerebellum, brain stem and spinal cord. Both peptides (10(-8) and 10(-7) M) caused a hyperpolarization of most astrocytes studied. The hyperpolarization by CCK was markedly reduced or blocked by the CCKB-antagonist L-365,260 whereas addition of the Bom-antagonist [D-Phe12,Leu14]-Bom antagonized the effects of Bom, suggesting a specific action of these peptides. When CCK and Bom were tested on the same cell, both peptides were effective, indicating a colocalization of receptors for CCK and Bom on the glial membrane. Our electrophysiological investigations provide strong evidence for the existence of functional CCK and Bom receptors on astrocytes.

Colocalization of cholinergic, adrenergic and peptidergic receptors on astrocytes.

By means of combined immunohistochemical and auto-radiographic techniques we have studied the colocalization of cholinergic, adrenergic and peptidergic binding sites on astrocytes in explant cultures of rat spinal cord, brain stem and cerebellum. Many astrocytes which were immunostained by the monoclonal muscarinic receptor antibody M 35 were also intensely labelled by 3H-noradrenaline, the beta-adrenergic antagonist 3H-dihydroalprenolol and the peptides 125I-angiotensin II, 125I-neuropeptide Y and 3H-bradykinin. Electrophysiological studies demonstrating that muscarine, the adrenergic agonists noradrenaline and isoprenaline as well as angiotensin II, neuropeptide Y and bradykinin affect the membrane potential of the same astrocytes provide further evidence for the coexistence of cholinergic, adrenergic and peptidergic receptors on astrocytes.

Autoradiographic localization of binding sites for neuropeptide Y and bradykinin on astrocytes.

The cellular localization of binding sites for the vasoactive peptides 125I-neuropeptide Y (NPY) and 3H-bradykinin (BK) was studied in explant cultures of rat cerebellum, brain stem and spinal cord by means of autoradiography. The majority of astrocytes in these cultures expressed binding of 125I-NPY and 3H-BK over their cell bodies and processes. Simultaneous staining of the cultures with anti-glial fibrillary acidic protein (GFAP) has shown that the labelled cells were GFAP-positive and could therefore be identified as astrocytes. In addition to glial cells, a great number of neurones also revealed binding sites for the neuropeptides. Our autoradiographic findings together with recent electrophysiological studies provide good evidence for the existence of NPY- and BK-receptors on astrocytes.

[Methods and suitability of treatment for dental anxiety]. / Een onderzoek naar methoden en doelmatigheid van angstbegeleiding.

211 Patients followed a special dental fear programme. The Tell-Show-Do Method was the method most often used. Half a year after having finished the programme all patients received a questionnaire. A majority (74%) of the 158 patients that returned the questionnaire reported a good contact with their home dentist.