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Biomedical subjects

M Witt

Publications and source records attributed to M Witt.

At least 73 records · Page 4Linked to original sources

The Kartagener syndrome--an electron microscopic studies of biopsied mucosa.

We studied 43 tissue samples taken from patients (16 male and 27 women) with clinical diagnosis of Kartagener's syndrome. The specimens were fixed and prepared for transmission electron microscopy. After preliminary light microscopic examination 23 cases were qualified for ultrastructural studies. We found abnormal number of cilia on the bronchial mucosa cells. The ciliary structure was abnormal. In several cases we observed lack of inner and/or outer dynein arms, abnormal location of central microtubules, and abnormal number of microtubular doublets. Other findings were as follows: replacement of ciliated cells by goblet ones, mucosal edema and inflammatory infiltrate.

Adolescent↗

GLP-1/GIP chimeric peptides define the structural requirements for specific ligand-receptor interaction of GLP-1.

The gastrointestinal hormones glucagon-like peptide-1 (GLP-1) and gastric inhibitory polypeptide (GIP) strongly stimulate insulin release. Despite their high N-terminal sequence similarity, GLP-1 does not bind to the GIP receptor and vice versa. To characterize the domains required for interaction of the peptide ligands with their specific receptors, we performed displacement studies with various synthetic GLP-1/GIP hybrid peptides on RINm5F insulinoma cells. Displacement of 125I-GIP and 125I-GLP-1 was measured using GLP-1/GIP chimeras which comprised GIP and GLP-1 sequences at different positions. The binding affinity to the GLP-1 receptor was found to be sensitive to GIP-like exchanges in the N-terminal 22 amino acids as well as in positions 13 and 15 (loss of affinity 280-fold to more than 1000-fold). C-terminal substitution of the GLP-1 sequence by GIP diminished the affinity towards the GLP-1 receptor only 20-fold. All hybrid peptides investigated showed minimal binding affinity for the GIP receptor, indicating that the entire GIP-sequence (1-31) is important for receptor recognition. These findings provide insight into the structural requirements for the specific interaction of two important insulinotropic peptides with their specific receptors.

Amino Acid Sequence↗

Embryonic and early fetal development of human taste buds: a transmission electron microscopical study.

BACKGROUND: Taste buds are assemblies of slender epithelial cells that receive chemical stimuli from the outer (oral) environment. In contrast to the large and well documented information on the morphology of taste buds in adult humans and animals, there are only a few reports on fetal ones, and ultrastructural studies of prenatal human taste buds are lacking completely. Therefore, the present investigation has been carried out to study the taste bud primordium, its morphological changes including synaptogenesis, cell differentiation, and taste pore formation from the time of the onset of taste bud formation around the 8th week until the 15th postovulatory week. METHODS: Taste bud primordia of 42 human embryonic/fetal tongues have been examined by means of transmission electron microscopy. RESULTS: Nerve fibers approach the lingual epithelium between the 6th and 7th postovulatory week. They penetrate the basal lamina during the 8th week and form synapses with poorly differentiated, elongated, epithelial cells. By the 12th week, more differentiated cell types are seen: 1) electron-dense cells resembling type III cells of the adult taste bud containing large numbers of dense-cored vesicles (80-150 nm in diameter); 2) electron-dark cells with well developed endoplasmic reticulum and many apical mitochondria, being candidates for type II cells. Basally, these cells have foot-like processes containing dense-cored vesicles (120-200 nm in diameter), but they do not synapse to nerve fibers. Type I cells, characterized by apically located dense secretory granules, are not observed. First shallow grooves above the taste bud primordium are found around the 10th week. Untypically differentiated apical cellular processes extend onto the surface. Most of the taste pores develop around the 14th to 15th week. In the taste pit, mucous material is not present during the first 15 weeks of gestation. Synapses between cells and afferent nerve fibers were found by the 8th week, reaching a maximum around the 12th to 13th week. CONCLUSIONS: The early presence of taste bud cells containing dense-cored vesicles suggests an at least dual function of embryonic/ fetal taste buds: First, from the 8th until the 14th week, non-gustatory, paracrine functions should be considered. After the 14th week of gestation, when typical taste pores are present, the taste buds possibly start their gustatory function. Differentiated marginal cells are possibly involved in the formation of the taste pore. The lack of type I cells producing the mucous material in the taste pit indicates that the taste bud has not achieved a fully developed function until the 15th week of gestation.

Aging↗

Multicenter evaluation of broth microdilution method for susceptibility testing of Cryptococcus neoformans against fluconazole.

We have developed a microdilution method for measuring the susceptibility of Cryptococcus neoformans to fluconazole. The present study evaluated the interlaboratory agreement of the results for the microdilution method obtained at three different sites and compared this method with the National Committee for Clinical Laboratory Standards M27-P reference method. Excellent interlaboratory agreement among the results obtained at the three sites was achieved with this method (83 and 96% agreement within 1 and 2 log2 dilutions, respectively). An overall agreement of 90% between the microdilution method and the M27-P method was observed, demonstrating the comparability of the two methods. However, there are inherent problems with the M27-P method in relation to measuring C. neoformans susceptibility, including suboptimal growth of the organism in RPMI 1640, a longer incubation period, and a narrow range of MICs. On the basis of these data, the microdilution method tested in this study is recommended for inclusion in the National Committee for Laboratory Standards method for testing the antifungal susceptibility of C. neoformans.

AIDS-Related Opportunistic Infections↗

Distribution of vasoactive intestinal peptide-like immunoreactivity in the taste organs of teleost fish and frog.

Using immunohistochemistry, vasoactive intestinal peptide (VIP) was visualized in taste bud cells of the carp, Cyprinus carpio, and the European catfish, Silurus glanis, by means of light and electron microscopy. Intracellular membrane systems, presumably smooth endoplasmic reticulum, of light (sensory) cells, but not of dark (supporting) cells and basal cells, were densely labelled with antibody. In the frog (four species: Rana temporaria, R. ridibunda, R. arvalis, R. pipiens), taste bud cells did not label. However, the dense basal nerve fibre plexus, some subepithelial ganglionic cells, but no ascending intragemmal fibres, were immunoreactive. In fish, the results support evidence that VIP is involved in the modulation of taste transduction at the level of receptor cells. In the frog, an indirect, possibly vasodilatatory effect on taste perception may be considered.

Animals↗

The correlation of cavernous systolic occlusion pressure with peak velocity flow using color duplex Doppler ultrasound.

This prospective study involved 27 patients who underwent dynamic infusion cavernosometry/cavernosography and color duplex Doppler ultrasound. Phase III dynamic infusion cavernosometry/cavernosography determination of cavernous artery systolic occlusion pressure and color duplex Doppler measurements of peak velocity flow were performed. Four subpopulations were defined: 1) normal cavernous artery systolic occlusion pressure and peak velocity flow, 2) abnormal systolic occlusion pressure and normal peak velocity flow, 3) normal systolic occlusion pressure and abnormal peak velocity flow, and 4) abnormal systolic occlusion pressure and peak velocity flow. Peak velocity flow significantly correlated with cavernous artery systolic occlusion pressure and, using a normal value of greater than 25 cm. per second, resulted in a sensitivity and specificity of 95%. The positive predictive value of an abnormal peak velocity flow was calculated to be 95%. We conclude that peak velocity flow is an effective, reliable and noninvasive means of evaluating corporeal arterial function in an office setting.

Adult↗

Structure/activity characterization of glucagon-like peptide-1.

Glucagon-like peptide-1 is a gastrointestinal hormone that strongly stimulates insulin release via specific receptors on the pancreatic beta-cell. To characterize the side-chain groups required for interaction of glucagon-like peptide-1 with its receptor, we performed binding studies with alanine-substituted glucagon-like peptide-1 analogues on RINm5F insulinoma cells. The binding affinity and biological activity of glucagon-like peptide-1 have been found to be sensitive to alanine exchanges in the N-terminal positions 1, 4, 6 and the C-terminal positions 22 and 23. Alanine substitutions at positions 5, 8, 10-12, 14, 16-21 and 25-30 do not change receptor affinity. These findings could be exploited to design glucagon-like peptide-1 agonists and probably antagonists for further physiological studies.

Amino Acid Sequence↗

Is sex worth searching for?

Several methods of molecular sexing (i.e. detection of Y-chromosomal sequences) have been briefly presented. In addition to other applications they can be used for monitoring of patients after sex-mismatched bone marrow transplantation for mixed chimerism and for detection of fetal cells in maternal circulation. Clinical significance of mixed chimerism and possibilities of non-invasive prenatal diagnosis with the use of fetal cells isolated from peripheral blood of pregnant women has been described.

DNA↗

Ultrastructure of the taste disc in the red-bellied toad Bombina orientalis (Discoglossidae, Salientia).

The taste disc of the red-bellied toad Bombina orientalis (Discoglossidae) has been investigated by light and electron microscopy and compared with that of Rana pipiens (Ranidae). Unlike the frog, B. orientalis possesses a disc-shaped tongue that cannot be ejected for capture of prey. The taste discs are located on the top of fungiform papillae. They are smaller than those in Ranidae, and are not surrounded by a ring of ciliated cells. Ultrastructurally, five types of cells can be identified (mucus cells, wing cells, sensory cells, and both Merkel cell-like basal cells and undifferentiated basal cells). Mucus cells are the main secretory cells of the taste disc and occupy most of the surface area. Their basal processes do not synapse on nerve fibers. Wing cells have sheet-like apical processes and envelop the mucus cells. They contain lysosomes and multivesicular bodies. Two types of sensory cells reach the surface of the taste disc; apically, they are distinguished by either a brush-like arrangement of microvilli or a rod-like protrusion. They are invaginated into lateral folds of mucus cells and wing cells. In contrast to the situation in R. pipiens, sensory cells of B. orientalis do not contain dark secretory granules in the perinuclear region. Synaptic connections occur between sensory cells (presynaptic sites) and nerve fibers. Merkel cell-like basal cells do not synapse onto sensory cells, but synapse-like connections exist between Merkel cell-like basal cells (presynaptic site) and nerve fibers.

Animals↗

A simplified method for detection of the mutations predominantly causing cystic fibrosis and phenylketonuria in Polish families.

Genomic DNA was isolated from dried blood specimens and subsequently used as a template in simplified PCR-based detection assays of delta F508 mutation of CFTR gene and of R408W mutation of PAH gene in families with cystic fibrosis and phenylketonuria, respectively. Products of amplification of CFTR gene were analyzed in NuSieve agarose gel. The amplification-created restriction site with TaqI digestion was used for detection of the PAH gene mutation.

Base Sequence↗

An improved, non-isotopic method of screening cells from patients with abnormalities of sexual differentiation for Y chromosomal DNA content.

The detection of 45,X/46,XY mosaicism in patients with abnormalities of sexual differentiation is of crucial diagnostic importance. Here we present application of a PCR based method of detection of alphoid repeats of Y chromosomal origin. The method detects 0.01% of male DNA on a female DNA background. Out of 28 patients studied, in all cases where the Y chromosome or a part of it containing centromeric sequences was present, a positive amplification signal of Y chromosomal alphoid repeats was detected. In five cases the Y origin of marker chromosomes was diagnosed. The pattern of amplification signal distribution of the SRY gene was identical to that of Y specific alphoid primers, which confirms applicability of this method in the molecular diagnostic laboratory. The other diagnostic advantage is the ability to use dried blood specimens as an easy to handle and efficient source of DNA.

DNA↗

Binding specificity and signal transduction of receptors for glucagon-like peptide-1(7-36)amide and gastric inhibitory polypeptide on RINm5F insulinoma cells.

Glucagon-like peptide-1(7-36)amide (GLP-1(7-36) amide) and gastric inhibitory polypeptide (GIP), peptides of the glucagon family, stimulate insulin secretion in vitro and in vivo. They possess high N-terminal sequence homology. Binding studies with 125I-labelled GIP and 125I-labelled GLP-1(7-36)amide were performed in RINm5F insulinoma cells to investigate receptor specificity and to compare both receptors directly. Both binding sites were highly ligand-specific: GIP did not bind to the GLP-1(7-36)amide receptor and vice versa. Both peptides increased intracellular cyclic AMP levels; GLP-1(7-36)amide was 100-fold more potent in stimulating cyclic AMP production when compared with GIP. At ranges of 1-10 nmol GLP-1(7-36)amide/l and 0.1-10 nmol GIP/l, corresponding to submaximal binding concentrations, the hormones showed an additive effect on cyclic AMP production. The N-terminal portion of GIP was important for binding, as GIP(1-30) showed almost full binding and biological activity. GIP(17-42) bound in a concentration-dependent manner with approximately 500-fold lower potency than GIP. At concentrations of up to 10 mumol GIP(17-42)/l no stimulation of cyclic AMP was observed.

Animals↗

[Principles of muscle efficiency in weightlessness].

Depending on the time spent in a space station, weightlessness causes varying changes to various organs and regulatory mechanisms of the human organism, among them the structures of the active and passive locomotor system. Experience has shown that regular physical activity using varying forms of training is necessary to keep astronauts and cosmonauts healthy, efficient and fit for work and to prepare them for a safe return to terrestrial conditions. One of the main tasks of physical activity is therefore to counteract the changes in the neuromuscular function, muscle strength, endurance and fatigability under conditions of weightlessness. So far, the diagnostics of these changes was limited to dynamometric measurements before and after space flights as no dynamometric method had been devised which was precise enough under conditions of weightlessness. The development and the introduction of the dynamoergometer Motomir is described against the background of previous results obtained by dynamometric, ergometric, morphometric, biochemical, histochemical and neuromuscular examinations before and after missions of varying duration under conditions of weightlessness. This device was used for the first valid and reproducible examinations of muscle strength and endurance aboard the space station Mir during the Austro-Russian space mission Austromir. The results of the examinations with this speed-controlled dynamometer and training device obtained during a short-term flight of 10 days and a long-term flight of 6 months are described and compared to previous results in the literature. Based on the existing training devices, training methods and training plans and the results of the Motomir Study, plans for devices and methodologies for future long-term space flights of up to 3 years' duration will be discussed. Finally, the possible use of a speed-controlled dynamometer and training device for rehabilitation on earth after injuries, surgery and immobilization of the active and passive locomotor system as well as in the bed-rest syndrome is being presented.

Animals↗

Ullrich-Turner syndrome with a small ring X chromosome and presence of mental retardation.

Since some patients with Ullrich-Turner syndrome (UTS) have mental retardation, we reviewed our experience to look for a high-risk subgroup. Among 190 UTS and gonadal dysgenesis patients with X chromosome abnormalities, 12 had mental retardation. All of the six (100%) with a small ring X were educable (EMI) or trainable mentally impaired (TMI) with more severe delay than expected in UTS. Among the 184 with other X abnormalities, only 6 had similar delays (2 from postnatal catastrophes), for a frequency of 3.3% mental retardation among those without a small ring X; only 2.2% of these had unexplained mental retardation. Polymerase chain reaction studies showed no Y-derived material in the 2 patients who were evaluated, and in situ hybridization confirmed X origin of the ring in the 6 subjects who were evaluated. We describe the phenotype of the 6 individuals with a small ring X, and an additional 2 patients with a small ring X who were identified outside the survey. The subjects with a small ring X comprised a clinically distinct subgroup which had EMI/TMI and shorter stature than expected in UTS. Seizures and a head circumference less than 10th centile were observed in half of the patients with a small ring X, and strabismus, epicanthus, and single palmar creases were present in more than half. A "triangular" face in childhood, pigmentary dysplasia, sacral dimple, and heart defects were also common. Neck webbing appeared to be less frequent than in 45,X. We hypothesize that the high risk of mental retardation in this form of the UTS results from lack of lyonization of the ring X due to loss of the X inactivation center. Excluding those with a small ring X, mental retardation is not significantly increased in patients with UTS.

Adolescent↗

Comparative lectin histochemistry on taste buds in foliate, circumvallate and fungiform papillae of the rabbit tongue.

Taste buds (TB) in the foliate, circumvallate and fungiform papillae of the rabbit tongue were examined with lectin histochemistry by means of light (LM) and electron (EM) microscopy. Biotin- and gold-labeled lectins were used for the detection of carbohydrate residues in TB cells and subcutaneous salivary glands. At the LM level, the lectins of soybean (SBA) and peanut (PNA) react with material of the foliate and circumvallate taste pores only after pretreatment of the section with neuraminidase. This indicates that the terminal trisaccharide sequences are as follows: Sialic acid-Gal-GalNAc in O-glycosylated glycoproteins or Sialic acid-Gal-GlcNAc in N-glycosylated glycoproteins. In fungi-form taste buds the lectins of Dolichos biflorus (DBA) and Helix pomatia (HPA), also specific to GalNAc residues, are reactive without preincubation with neuraminidase. Wheat germ agglutinin (WGA), specific to GlcNAc, reacts with TBs of all papillae; and the lectin from Ulex europaeus (UEA I), specific to fucose, binds to individual TB cells. The presence of sialic acid may protect mucus or other glycoproteins in TB cells and inside the taste pore from premature enzymatic degradation. In a post-embedding EM procedure on LR-White-embedded tissue sections, only gold-labeled HPA was found to bind especially on membrane surfaces of the microvilli which protrude into the taste pore; however HPA did not bind to the electron-dense mucus inside the taste pore. The mucus situated in the trough and at the top of the adjacent epithelial cells also is strongly HPA-positive, but is of different origin and composition than that found in the taste pore. These results demonstrate distinct carbohydrate histochemical differences between fungiform and circumvallate/foliate taste buds. The different configuration of galactosyl residues and the occurrence of mannose in circumvallate and foliate TBs leads to the suggestion that the lectin reactivities of TBs are not only due to the presence of mucins, but also to N-linked glycoproteins, possibly with a hormone-like paraneuronal function. A possible relationship to v. Ebner glands in these papillae is discussed.

Animals↗