Biomedical subjects

A de Haan

Publications and source records attributed to A de Haan.

At least 73 records · Page 4Linked to original sources

PubMed1997

Rapid antibody test for diagnosing fragile X syndrome: a validation of the technique.

To date, the identification of patients and carriers of the fragile X syndrome has been carried out by DNA analysis by means of the polymerase chain reaction and Southern blot analysis. This direct DNA analysis allows both the size of the CGG repeat and methylation status of the FMR1 gene to be determined. We have recently presented a rapid antibody test on blood smears based on the presence of FMRP, the protein product of the FMR1 gene, in lymphocytes from normal individuals and the absence of FMRP in lymphocytes from patients. Here, we have tested the diagnostic value of this new technique by studying FMRP expression in 173 blood smears from normal individuals and fragile X patients. The diagnostic power of the antibody test is "perfect" for males, whereas the results are less specific for females.

Antibodies, Monoclonal↗

PubMed1997

Use of gene targeting for compromising energy homeostasis in neuro-muscular tissues: the role of sarcomeric mitochondrial creatine kinase.

We have introduced a single knock-out mutation in the mitochondrial creatine kinase gene (ScCKmit) in the mouse germ line via targeted mutagenesis in mouse embryonic stem (ES) cells. Surprisingly, ScCKmit -/- muscles, unlike muscles of mice with a deficiency of cytosolic M-type creatine kinase (M-CK -/-; Van Deursen et al. (1993) Cell 74, 621-631), display no altered morphology, performance or oxidative phosphorylation capacity. Also, the levels of high energy phosphate metabolites were essentially unaltered in ScCKmit mutants. Our results challenge some of the present concepts about the strict coupling between CKmit function and aerobic respiration.

Adenosine Triphosphate↗

PubMed1996

Myosin heavy chain isoform expression and high energy phosphate content in human muscle fibres at rest and post-exercise.

1. The relationship between myosin heavy chain (MyHC) isoforms and high energy phosphate content was studied in human muscle fibres at rest and following maximal dynamic exercise lasting 25 s. 2. Single fibre fragments were characterized as type I, type IIA, type IIX or type IIAX. These latter fibres were subdivided into five groups on the basis of the proportion of MyHC IIX isoform present. 3. Resting ATP concentration in type I fibres was 10% lower than in type II fibres (P < 0.05), but no differences were found amongst type IIA, IIX and IIAX fibre groups. Phosphocreatine (PCr) content was lower in type I than in type II fibres (P < 0.01) and, amongst type II fibres, increased progressively with the amounts of MyHC IIX expressed. 4. After 25 s of maximal dynamic exercise ATP concentration was reduced in all fibres. The decrease was approximately 25% in type I fibres and between 47 and 66% in the type II subgroups. 5. Post-exercise PCr content was low in all fibre types. Fibre groups with the lowest post-exercise PCr also had the lowest ATP and the highest inosine monophosphate contents. delta PCr (rest to post-exercise) was smallest in type I fibres and showed a progressive increase in the type II fibre groups as the proportion of the faster IIX myosin heavy chain isoform increased.

Adenosine Triphosphate↗

PubMed1996

Activation of murine lymphocytes by lipopolysaccharide incorporated in fusogenic, reconstituted influenza virus envelopes (virosomes).

We have studied the in vitro activation of murine lymphocytes with LPS incorporated in the membranes of both phospholipid vesicles (liposomes) and vesicles composed of fusogenic, reconstituted influenza virus envelopes (virosomes). The incorporation of Salmonella minnesota rough-LPS in liposomes reduced the potency of LPS to stimulate splenocyte proliferation and cell surface kappa-light chain expression on 70 Z/3 pre-B cells by over 100-fold. Salmonella minnesota rough-LPS inserted into virosomes was at least 10-fold more potent than free LPS, both when prebound virosomes were allowed to be taken up by the cells at neutral pH and when the virosomes were fused into the plasma membrane by low pH treatment. Inactivation of the virosomes by low pH pretreatment reduced the potency of the virosomal LPS to the level of liposome-incorporated LPS. The association of the various LPS forms with the cells was quantitated using radio-iodinated LPS. Correcting for uptake, virosomal LPS remained 2- to 10-fold more potent than free LPS in stimulating B lymphocytes and at least 100-fold more active than liposomal LPS or fusion-inactivated virosomes. After low pH-induced fusion with the plasma membrane, the majority (80%) of the prebound virosomes had fused with the cells, compared with about 8% after neutral uptake. From these results we conclude that LPS inserted into the plasma or endosomal membranes efficiently activates murine lymphocytes. The fusion data suggest that the incorporation into endosomal membranes might be a more effective stimulus.

Animals↗

PubMed1996

The sequence of a small subunit of cytochrome c oxidase from Crithidia fasciculata which is homologous to mammalian subunit IV.

The sequence of subunit 8 of cytochrome c oxidase from Crithidia fasciculata was determined by sequencing cDNA and N-terminus of the mature protein (Mr = 15.7 kDA). The (inferred) protein is homologous to mammalian cox IV and the corresponding cox subunits from yeast, Neurospora crassa and Dictyostelium discoideum, which is reflected in a very similar hydropathy profile. Elements that are conserved in the C. fasciculata sequence include (i) an N-terminal (D/E)-(K/R)-X-K-(X2)-W-(X2)-(I/L) motif, (ii) a putative membrane-spanning region in the middle portion of the protein, and (iii) a C-terminal W-(X13)-(N/D)-P motif. The C. fasciculata protein is synthesized with a cleavable presequence.

Amino Acid Sequence↗

PubMed1996

Absence of an effect of fatigue on muscle efficiency during high-intensity exercise in rat skeletal muscle.

The effect of fatigue was studied on rat skeletal muscle efficiency during maximal dynamic exercise of 10s duration. After the initial 4s of exercise, power output decreased rapidly to 46.2 +/- 6.7% (mean +/- SD; n = 6) after 6s of stimulation and further to 17.5 +/- 5.8% in the last contraction. Both the rates of total work output and high-energy phosphate consumption decreased with increasing exercise duration. As a result muscle efficiency was not affected by exercise time in the present experiments. This result indicates that fatigue in severe maximal exercise is induced by a feed-back mechanism, which in the case of high ATP utilisation rates will reduce ATP splitting probably by reducing Ca(2+)-release from the sarcoplasmic reticulum.

Animals↗

PubMed1996

Purification and characterization of cytochrome c oxidase from the insect trypanosomatid Crithidia fasciculata.

Cytochrome c oxidase was purified from the mitochondrial lysate of the insect trypanosomatid Crithidia fasciculata with the aid of a methyl hydrophobic interaction column in a rapid one-step procedure. The purified complex displayed all characteristics expected from a eukaryotic cytochrome c oxidase: the presence of CuA in electron paramagnetic resonance analysis, a characteristic 605 nm peak in reduced-minus-oxidized optical spectroscopy, and the capacity to efficiently oxidize homologous, but not heterologous, cytochrome c. Two-dimensional PAGE showed that C. fasciculata cytochrome c oxidase consists of at least 10 different subunits. N-terminal sequences were obtained from the six smallest subunits of the complex, one of them showing significant similarity to Neurospora crassa cytochrome c oxidase subunit V. The N-terminus of each of the four largest subunits was found to be blocked.

Amino Acid Sequence↗

PubMed1996

Mucosal immunoadjuvant activity of liposomes: role of alveolar macrophages.

Previously, we have reported on a liposomal adjuvant system for stimulation of both systemic IgG and mucosal s-IgA responses against viral antigens (influenza virus subunit antigen or whole inactivated measles virus) administered intranasally to mice. Immune stimulation is observed with negatively charged, but not with zwitterionic, liposomes and is independent of a physical association of the antigen with the liposomes. Furthermore, liposome-mediated immune stimulation requires deposition of the liposomes and the antigen in the lower respiratory tract. In the present study, it is shown that alveolar macrophages (AM) are the main target cells for negatively charged liposomes administered to the lungs of mice. AM isolated from animals, to which negatively charged liposomes were administered beforehand, showed large intracellular vacuoles, suggestive of massive liposome uptake. Under ex vivo conditions, both AM and RAW 264 cells exhibited a high capacity to take up negatively charged liposomes. The deposition of negatively charged liposomes, but not zwitterionic, liposomes in the lung reduced the phagocytic and migratory behaviour of AM, as assessed on the basis of transport of carbon particles to the draining lymph nodes of the lungs. Depletion of AM in vivo with dichloromethylene diphosphonate, facilitated an enhanced systemic and local antibody response against influenza subunit antigen deposited subsequently to the lower respiratory tract. In conclusion, these data provide support for the hypothesis that uptake of negatively charged liposomes blocks the immunosuppressive activity of AM, thereby facilitating local and systemic antibody responses.

Adjuvants, Immunologic↗

PubMed1996

In vivo IIX and IIB fiber recruitment in gastrocnemius muscle of the rat is compartment related.

The purpose of the present study was to investigate to what extent fast-twitch IIX and IIB fiber recruitment was related to the natural existing muscle compartments (subvolumes of muscle innervated by different primary nerve branches) in rat medial gastrocnemius. Three groups (n = 6) of rats trotted on a motor-driven treadmill (20 degrees incline) at different speeds. A fourth group served as controls, and a fifth group received in situ electrical stimulation of all medial gastrocnemius muscle fibers. Postexercise glycogen levels (periodic acid-Schiff staining intensities) were made. Running caused more and in situ stimulation caused less glycogen breakdown in the proximal IIX and IIB fibers compared with the fibers of the same type in the most distal compartment. Furthermore, the boundaries of the most distal compartment could often be recognized in the periodic acid-Schiff-stained cross sections. It was concluded that during running the proximal IIX and IIB fibers were recruited to a greater extent (and at lower treadmill speeds) compared with the distal IIX and IIB fibers, respectively.

Animals↗

PubMed1995

Induction of a secretory IgA response in the murine female urogenital tract by immunization of the lungs with liposome-supplemented viral subunit antigen.

This study demonstrates that liposomes administered to the lower respiratory tract of mice have the capacity to stimulate secretory IgA (s-IgA) antibody production in the female urogenital system. Total respiratory tract immunization of mice with influenza virus subunit antigen simply mixed with negatively charged liposomes induced antigen-specific s-IgA in vaginal secretions, in addition to systemic IgG and s-IgA in the respiratory tract. Immunization of the upper respiratory tract alone or oral immunization were ineffective. These observations demonstrate that, upon stimulation with liposomes, the lymphoid tissue associated with the lung can act as an inductive site for migration of IgA-committed B cells to distant mucosal tissues, including the female urogenital tract. It is concluded that liposomes administered to the lower respiratory tract provide a promising adjuvant system for stimulation of both systemic and mucosal antibody responses against coadministered antigen, including production of s-IgA at distant mucosal sites.

Adjuvants, Immunologic↗

PubMed1995

Liposomes as an immunoadjuvant system for stimulation of mucosal and systemic antibody responses against inactivated measles virus administered intranasally to mice.

This paper reports on the immune-stimulatory activity of liposomes in an inactivated whole measles virus vaccine preparation administered intranasally to mice. Liposomes, simply mixed with inactivated whole measles virus, significantly stimulated the serum IgG response relative to the response to the virus alone. In addition, the liposomal vaccine, but not the free virus, induced a secretory IgA (s-IgA) response in the lungs and nasal cavity. Serum IgG and s-IgA responses persisted up to at least 24 weeks post-immunization. The liposomes induced a moderate increase in the serum IgG response, but no s-IgA response, following intramuscular immunization. It is concluded that liposomes provide a promising adjuvant system for induction of high systemic as well as mucosal antibody responses against inactivated measles virus in an intranasal or inhalation vaccine formulation.

Adjuvants, Immunologic↗

PubMed1995

Mucosal immunoadjuvant activity of liposomes: induction of systemic IgG and secretory IgA responses in mice by intranasal immunization with an influenza subunit vaccine and coadministered liposomes.

This paper reports on a novel immunoadjuvant activity of liposomes. An influenza subunit preparation, containing the isolated viral surface antigens, was incorporated in a liposomal formulation. Administration of this vaccine to mice via the intranasal (i.n.) route resulted in a stimulated serum IgG response relative to the response to i.n. immunization with the antigen alone. In addition, the liposomal vaccine induced a secretory IgA (sIgA) response in the mucosa of the lungs and nasal cavity. Both serum IgG and sIgA responses persisted up to at least 21 weeks postimmunization. Immune stimulation was observed with negatively charged liposomes consisting of phosphatidylcholine (PC), cholesterol and dicetylphosphate (DCP), but not with zwitterionic liposomes, consisting of PC and cholesterol alone. Remarkably, for stimulation of serum IgG responses and induction of an sIgA response, liposomes could be simply mixed with the antigen. Moreover, i.n. administration of empty liposomes up to 48 h prior to i.n. immunization with the subunit antigen also resulted in immune stimulation, indicating that the liposomes did not exert their adjuvant effect by acting as a carrier for the antigen. The liposomal vaccine conferred protection against infection. It is concluded that liposomes, administered i.n., provide a promising adjuvant system for stimulation of antibody responses in general, and mucosal sIgA responses in particular.

Adjuvants, Immunologic↗

PubMed1995

The effect of different dosages of caffeine on endurance performance time.

The effect of different dosages of caffeine (0-5-9-13 mg.kg body weight-1) on endurance performance was examined. Nine well-trained cyclists participated in this study (VO2max 65.1 +/- 2.6 ml.kg-1.min-1). Caffeine capsules were administered in random order and double-blind. One hour after capsule ingestion, subjects cycled until exhaustion at 80% Wmax on an electromagnetically braked cycle ergometer. Blood samples were taken before, during and after the exercise test. Before and after the test a urine sample was obtained. A significant increase in endurance performance was found for all caffeine tests compared to placebo (endurance time 47 +/- 13, 58 +/- 11, 59 +/- 12 and 58 +/- 12 min for 0, 5, 9 and 13 mg.kg-1 body weight, respectively). No differences were found in endurance performance between the three caffeine dosages which indicates that no dose-response relation of caffeine and endurance performance was found. An increased free fatty acid and glycerol concentration was found after caffeine consumption compared with placebo. The mean urinary caffeine concentrations after exercise were 4.8 +/- 1.8, 8.9 +/- 5.2 and 14.9 +/- 6.9 micrograms.ml-1 urine for 5, 9 and 13 mg of caffeine.kg-1 body weight. Only the lowest dose of caffeine resulted in urine caffeine concentrations below the doping limit of the International Olympic Committee of 12 micrograms.ml-1 urine in all individuals. It is concluded that caffeine is an ergogenic aid that stimulates endurance performance. A dose-response relation between caffeine and endurance time was not found for the dose-range investigated.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

PubMed1995

The effects of MM-creatine kinase deficiency on sustained force production of mouse fast skeletal muscle.

Twitch force production was normal in muscles of mice which lack MM-creatine kinase, but the tetanic force:twitch force ratio was lower than in control muscles (3.60 vs 4.27; P < 0.05). In a series of repeated tetanic contractions the force in the second contraction was already markedly depressed (20-50%), while subsequently only small changes were observed. The effect was greater in exercise that would require a higher metabolic peak flux. The depressed force production was not accompanied by a slowing of relaxation, indicating that enough ATP was present to sustain myofibrillar ATPase and Ca(2+)-ATPase activity.

Adenosine Triphosphatases↗

PubMed1995

Beta-adrenoceptor blockade and skeletal muscle energy metabolism during endurance exercise.

Twelve healthy male volunteers cycled to exhaustion at a workload corresponding to 70% of maximal aerobic power after administration of 80 mg of the beta 1+2-adrenoceptor antagonist propranolol and after administration of placebo by mouth. Exercise times until exhaustion were 39 +/- 7 and 86 +/- 7 min in the propranolol and placebo groups, respectively. Muscle inosine 5'-monophosphate content was significantly increased above resting levels at exhaustion after placebo. At exhaustion after propranolol, inosine 5'-monophosphate was not increased significantly and was lower than at exhaustion after placebo. No changes in ATP and the total adenine nucleotide content during exercise were found in the two tests. Muscle glycogen content was significantly reduced at exhaustion after placebo as well as after propranolol, but the levels were still significantly higher at exhaustion after propranolol than after placebo. No evidence for a shift in glycogen utilization among types I, IIa, and IIb fibers after propranolol was found. The results show that neither an imbalance between ATP utilization and ATP regeneration nor premature glycogen depletion, either in the whole muscle or in specific muscle fiber types, provides a satisfactory explanation for the premature fatigue during endurance exercise after propranolol.

Adenine Nucleotides↗

PubMed1995

The mATPase histochemical profile of rat type IIX fibres: correlation with myosin heavy chain immunolabelling.

In the present study we report a novel histochemical method which, by sequential pre-incubations in alkaline and acidic media, selectively differentiates muscle fibres expressing myosin heavy chain IIX, on the basis of a specific profile for myofibrillar actomyosin ATPase (mATPase) activity. The enzyme reactions were tested for specificity by means of anti-myosin heavy chain monoclonal antibodies, which were characterized on Western blots of muscle homogenates. Enzyme histochemical reactions with the traditional pH buffers were compared to those of the new method and, in conjunction with the immunoreactions, used to confirm the relationship between MyHC expression and the distinct profiles for mATPase. Immunohistochemical reactions demonstrated that the new method only differentiates those fibres expressing myosin heavy chain IIX. The method revealed a continuum in which the intermediate staining intensities corresponded to hybrid fibres expressing myosin heavy chain IIX in combination with either the IIA or IIB forms. Quantitative histochemistry and immunohistochemistry (by image analysis), used to examine the relationship between staining intensities for mATPase and amounts of myosin heavy chain IIX expression, revealed that the new method discriminates well between hybrid fibres expressing variable amounts of the IIX isoform (r2 = 0.93).

Animals↗

PubMed1994

Novel pattern of editing regions in mitochondrial transcripts of the cryptobiid Trypanoplasma borreli.

In mitochondria of Kinetoplastida belonging to the suborder Trypanosomatina, the nucleotide sequence of transcripts is post-transcriptionally edited via insertion and deletion of uridylate residues. In order to shed more light on the evolutionary history of this process we have searched for editing in mitochondrial RNAs of Trypanoplasma borreli, an organism belonging to the suborder Bodonina. We have cloned and sequenced a 5.3 kb fragment derived from a 37 kb mitochondrial DNA molecule which does not appear to be a part of a network structure and have found genes encoding cytochrome c oxidase (cox) subunit 1, cox 2 and apocytochrome (cyt) b, and genes encoding the small and large subunit mitoribosomal RNAs. The order in which these genes occur is completely different from that of trypanosomatid maxicircle genes. The 5' and 3' termini of both the cytb and cox1 gene are cryptic, the protein coding sequences being created by extensive insertion/deletion of Us in the corresponding mRNA sections. Phylogenetic analyses of the protein and ribosomal RNA sequences demonstrated that the separation between T.borreli and Trypanosomatina was an early event, implying that U-insertion/deletion processes are ancient. Different patterns of editing have persisted in different lineages, however, since editing of cox1 RNA and of relatively small 3'-terminal RNA sections is not found in trypanosomatids. In contrast, cox2 RNA which is edited in trypanosomatids by the insertion of four Us, is unedited in T.borreli.

Amino Acid Sequence↗

PubMed1994

Prediction of mental status in carriers of the fragile X mutation using CGG repeat length.

For genetic counseling in fragile X (fra(X)) families, it is important to know the diagnostic impact of the CGG repeat length in carriers of the fragile X mutation. We have analyzed the CGG repeat length in 106 males and 73 females who had inherited the maternal fra(X) mutation. The sensitivity, specificity, and predictive value of the CGG repeat analyses, as measured on Southern blots of PstI/EcoRI digests, were calculated. In males the sensitivity, specificity and negative predictive value were 99%, 100% and 94%; respectively. In females the specificity (60%) and, consequently, the positive predictive value (82%) was reduced. Therefore, it remains impossible to predict accurately whether a female fetus demonstrating a full mutation will be affected. On the other hand, the negative predictive value of the CGG test in females was 100%. We have no evidence, as yet, that for female carriers with the full mutation, cytogenetic analysis is of additional value to predict the mental status. However, we have observed one mentally retarded fra(X) case with extreme mosaicism in which a typical premutation fragment was the predominant DNA species. Therefore, until more data and better DNA tests are available, we would like to advocate additional cytogenetic investigation if in a fetus a CGG repeat length in the premutation range is found.

Blotting, Southern↗