Biomedical subjects

Andreas Müller

Publications and source records attributed to Andreas Müller.

53 records · Page 3Linked to original sources

PubMed2002

Hydrogen bond vibrations of 2-aminopyridine.2-pyridone, a Watson-Crick analogue of adenine.uracil.

The 2-aminopyridine.2-pyridone complex (2AP.2PY) is linked by antiparallel N-H.O=C and N-H.N hydrogen bonds, providing a model for the Watson-Crick hydrogen bond configuration of the adenine-thymine and adenine-uracil nucleobase pairs. Mass-selected S(1) <--> S(0) vibronic spectra of the supersonically cooled 2AP.2PY base pair analogue were measured by laser resonant two-photon ionization and emission spectroscopies. The hydrogen bond vibrations nu(2) (buckle, out-of-plane) and the three in-plane vibrations nu(3) (opening), nu(5) (shear), and nu(6) (stretch) were observed in the S(0) and S(1) states, giving detailed information on the stretching and deformation force constants of the (amide)N-H.N(pyridine) and the (amino)N-H.O=C hydrogen bonds. Density functional calculations with the B3LYP functional and the 6-311++G(d,p) and 6-311++G(2d,2p) basis sets yield ground-state hydrogen bond frequencies in close agreement with experiment.

Adenine↗

PubMed2002

Analysis of histone acetyltransferase and histone deacetylase families of Arabidopsis thaliana suggests functional diversification of chromatin modification among multicellular eukaryotes.

Sequence similarity and profile searching tools were used to analyze the genome sequences of Arabidopsis thaliana, Saccharomyces cerevisiae, Schizosaccharomyces pombe, Caenorhabditis elegans and Drosophila melanogaster for genes encoding three families of histone deacetylase (HDAC) proteins and three families of histone acetyltransferase (HAT) proteins. Plants, animals and fungi were found to have a single member of each of three subfamilies of the GNAT family of HATs, suggesting conservation of these functions. However, major differences were found with respect to sizes of gene families and multi-domain protein structures within other families of HATs and HDACs, indicating substantial evolutionary diversification. Phylogenetic analysis identified a new class of HDACs within the RPD3/HDA1 family that is represented only in plants and animals. A similar analysis of the plant-specific HD2 family of HDACs suggests a duplication event early in dicot evolution, followed by further diversification in the lineage leading to Arabidopsis. Of three major classes of SIR2-type HDACs that are found in animals, fungi have representatives only in one class, whereas plants have representatives only in the other two. Plants possess five CREB-binding protein (CBP)-type HATs compared with one to two in animals and none in fungi. Domain and phylogenetic analyses of the CBP family proteins showed that this family has evolved three distinct types of CBPs in plants. The domain architecture of CBP and TAF(II)250 families of HATs show significant differences between plants and animals, most notably with respect to bromodomain occurrence and their number. Bromodomain-containing proteins in Arabidopsis differ strikingly from animal bromodomain proteins with respect to the numbers of bromodomains and the other types of domains that are present. The substantial diversification of HATs and HDACs that has occurred since the divergence of plants, animals and fungi suggests a surprising degree of evolutionary plasticity and functional diversification in these core chromatin components.

Acetyltransferases↗

PubMed2002

Upper airway epithelial structural changes in obstructive sleep-disordered breathing.

The etiology of upper airway collapsibility in patients with snoring and obstructive sleep apnea (OSA) remains unclear. Structural mucosal changes could be contributory factors. The objective of this study was to determine whether pathologic changes in the epithelium or the epithelial-connective tissue interface are present in patients with snoring and/or OSA by means of scanning electron microscopy and immunohistochemistry. Uvulae were obtained by uvulopalatopharyngoplasty from three patients with habitual snoring and nine patients with mild to severe OSA, as well as by dissection from 43 nonsnoring body donors. Scanning electron microscopy revealed structural changes in the epithelial-connective tissue boundary that significantly differed from age-related changes in the control subjects. The immunohistochemical staining with antibodies against epithelial cytokeratins showed differences in the expression pattern of cytokeratin 13 between patients and control subjects. No differences were found in the distribution pattern of laminin. Analysis of defense cells revealed a significant diffuse infiltration of leukocytes, mainly T cells, inside the lamina propria of the patient group, which was not observed in the control group. In conclusion, these results support the hypothesis that progressive structural changes in the mucosa caused by the trauma of snoring are a possible contributory factor to upper airway collapsibility.

Adult↗

PubMed2002

Validated capillary gas chromatographic-mass spectrometric assay to determine 2-methylcitric acid I and II levels in human serum by using a pulsed splitless injection procedure.

BACKGROUND: Despite some clinical applications of 2-methylcitric acid (2-MCA) determination in urine and amniotic fluid, a diagnostic use of 2-MCA levels in serum is not common practice. This could be related to the complexity of the assay, or possibly to unawareness of other feasible clinical applications. METHODS: The levels of the diastereomers 2-MCA I and II in human serum were determined by GC-MS based on a method using a pulsed splitless injection technique. A stable isotope dilution principle was modified considering the diastereomer ratio and impurities of the internal standard. Precision parameters as well as recovery rates of the assay were determined. Reference intervals for 2-MCA(total), 2-MCA I and II levels were obtained in 52 healthy volunteers (31 female, 21 male, mean age 41.7+/-14.4 years). RESULTS: 2-MCA was readily detected in each sample of serum, as well as in urine, cerebrospinal fluid and amniotic fluid. The limit of detection was 10 nmol/l for 2-MCA(total). The internal standard showed a diastereomer ratio of 2-MCA II-d3 to 2-MCA I-d3 of 0.83+/-0.05, its chemical purity had to be corrected to 90.5+/-0.5%. In concentrations of 446, 750 and 1256 nmol/l 2-MCA(total), recovery rates of 98.5, 93.7 and 88% with a mean intra-assay RSD of 1.5% were determined. The day-to-day precision was 10% RSD (SD 40 nmol/l) for 2-MCA(total) obtained with a pooled serum sample at a concentration of 401 nmol/l 2-MCA(total) over a period of 5 months (n=17). The normal range for 2-MCA(total) in human serum was calculated as 81-266 nmol/l confirming previous findings. CONCLUSIONS: The GC-MS assay using a pulsed splitless injection procedure ensures a good response to differing concentrations of 2-MCA in various specimens. Considering exact determination of the diastereomer ratio as well as the purity of the internal standard, the assay offers good precision and recovery for 2-MCA I and II levels in serum.

Adult↗

PubMed2002

A case of chondronecrosis of the epiglottis after laser chordotomy.

A 30-year-old Caucasian female patient suffering from bilateral vocal-fold immobility developed a necrosis of the epiglottis with severe respiratory distress 35 days after fiber-guided Nd:YAG-laser chordotomy on the right side. Twenty-two years prior to surgery, she had received a high-dose radiotherapy, which had caused severe scarring of the whole neck and the larynx. This necrosis is more likely an inflammatory complication than a direct side effect of the laser surgery in the radiation-altered laryngeal tissue. It underlines the importance of carefully considering the indications for laser surgery of an irradiated larynx.

Adult↗

PubMed2002

Analysis of hypermethylation in the RPS element suggests a signal function for short inverted repeats in de novo methylation.

A repetitive DNA sequence (RPS) from Petunia hybrida had previously been shown to enhance expression variegation in petunia and tobacco and to carry a hot spot for de novo DNA methylation. Here we show that a strong de novo hypermethylation site is located within a palindromic segment of the RPS and present indirect evidence, based on sequence homologies to other repeat units within the RPS, for the formation of secondary structures at the methylation site in vivo. We demonstrate that the palindromic RPS element, which is moderately to highly repetitive in petunia, does not predominantly localise to constitutive heterochromatin. To test whether the RPS is subject to de novo methylation due to its repetitive nature or to intrinsic signals within the RPS, we integrated the RPS into the genome of Arabidopsis thaliana, a plant lacking homology to the RPS. Our data indicate that the palindromic element also acts as a de novo hypermethylation site in the non-repetitive genomic background of Arabidopsis, strongly suggesting a signal function of the palindromic RPS element.

Animals↗

PubMed2002

A retrospective pilot study to assess the impact of strabismus surgery on the corneal endothelium in children.

PURPOSE: To investigate if strabismus surgery has an impact on the corneal endothelium in children. METHODS: Fourteen children who had undergone uncomplicated strabismus surgery an average of 41 months previously (range 24-73 months) were compared with a group of age-matched (range 5-12 years) and refractive error (RE)-matched (range -4.74 to +4.25 D) children. Following noncontact specular microscopy, areas of at least 100 cells were measured, the number of sides for each cell recorded, and endothelial cell density (ECD) calculated from the area measures. Corneal diameter was measured with a clinical rule, corneal curvature with a manual keratometer, central corneal thickness (CCT) by ultrasound pachometry, and IOP by non-contact tonometry. RESULTS: The average ECD (+/-S.D.) in the post-surgery group was 4467+/-627 cells mm(-2), which was significantly higher (p < 0.02) than in the control group (3781+/-452 cells mm(-2)). This was because the average endothelial cell area in the surgery group was significantly smaller than in the control group (p < 0.005; 252+/-25 microm2 vs 296+/-29 microm2). Differences in cell areas were slightly greater for smaller cells (-15.7% for five-sided cells, -15.6% for six-sided cells, -14.3% for seven-sided cells and -13.8% for eight-sided cells). The cell differences appear to be related to both a slightly smaller corneal diameter (11.00+/-0.52 mm vs. 11.21+/-0.58 mm) and a slightly higher CCT (0.528+/-0.033 mm vs. 0.515+/-0.032 mm) in the post-operative group. CONCLUSION: The endothelial mosaic in children after strabismus surgery can show a higher than expected ECD value and slight cell pleomorphism. These signs indicate that there might be differences in eye growth and endothelial inflammatory stress associated with uneventful strabismus surgery.

Age Distribution↗

PubMed2002

Central corneal thickness in European (white) individuals, especially children and the elderly, and assessment of its possible importance in clinical measures of intra-ocular pressure.

PURPOSE: To ascertain the impact of central corneal thickness (CCT, as assessed by pachometry) and central corneal curvature (as assessed by keratometry) on clinical measures of intra-ocular pressure (IOP, as assessed by tonometry), especially in the young and elderly. METHODS: Pachometry, keratometry and tonometry were carried out on three groups, namely children aged 5-15 years, adults aged 32-60 years, and elderly individuals aged between 61 and 82 years. For children, ultrasound pachometry was combined with non-contact tonometry (NCT), specular microscopy was used with Perkins tonometry in the adults, and ultrasound pachometry was used with Perkins tonometry for the elderly. Central corneal curvature was assessed by keratometry. RESULTS: The average CCT in children was 0.529+/-0.034 mm (n = 104, +/-S.D.), averaged 0.533+/-0.033 in adults (n=75) and 0.527+/-0.034 mm (n=91) in the elderly. Tonometry values averaged 16.7+/-2.9 mm in children, 13.0+/-3.5 mmHg in adults and 13.6+/-2.5 mm in the elderly group. Central corneal thickness values were not predictably different in relation to central corneal curvature values. Regression analyses indicated that the tonometry values were higher in both children and the elderly who had thicker corneas (and vice versa) (p < 0.003), with the measures increasing by 1.3+/-0.4 and 2.6+/-0.4 mmHg for a 10% difference in CCT in children and the elderly, respectively. For adults, no statistically significant difference in tonometry values could be demonstrated with respect to CCT (<1 mmHg for a 10% difference in CCT), and for no group were the CCT or tonometry values predictably different in relation to central comeal curvature values. CONCLUSIONS: The results of these studies, albeit relatively small scale, indicate that in young or elderly individuals with essentially normal IOP and CCT measures, the tonometry values show only small differences with respect to CCT. The slope in the observed relationship was not that different from an average of 1.5 mmHg for a 10% different in CCT, as obtained from a literature analysis over a 30-year period. The magnitude of the effect does not provide evidence that pachometry needs to be routinely performed in glaucoma screening protocols based on tonometry. Notwithstanding, the finding of higher than expected tonometry values should be further investigated, by pachometry, especially in very young children and in the elderly.

Adolescent↗

PubMed2002

Differential regulation of Th1-type and Th2-type cytokine profiles in pancreatic islets of C57BL/6 and BALB/c mice by multiple low doses of streptozotocin.

In the nonobese diabetic (NOD) mouse, the T helper (Th)1-type inflammatory cytokines interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha play a critical role in the development of type 1 diabetes, whereas the Th2-type anti-inflammatory cytokines interleukin (IL)-4 and IL-10 operate counterregulatory. There are no comprehensive analyses on cytokine profiles in the mouse model of diabetes induced with multiple low doses of streptozotocin (MLD-STZ). Therefore, we used islets to study ex vivo effects of MLD-STZ and in vitro effects of STZ on IFN-gamma, TNF-alpha, IL-4, and IL-10 on both levels of protein-producing cells and the mRNA expression, as well as the mRNA expression of the Th3-type cytokine transforming growth factor TGF-beta1. C57BL/6 and BALB/c mice of both genders were injected intraperitoneally with 40 mg/kg body wt STZ on five consecutive days and islets were isolated on day I and 3 after the fifth STZ-injection. Control mice received the solvent of STZ. In islets of C57BL/6 mice of both genders MLD-STZ similarly stimulated production of IFN-gamma and TNF-alpha, but significantly reduced IL-4 and IL-10 levels in male mice only. Opposite results were obtained in islets of BALB/c mice of both genders. Here, MLD-STZ markedly decreased the levels of IFN-gamma and TNF-alpha, but significantly increased the levels of IL-4 and IL-10. The functional results were in line with MLD-STZ effects on the mRNA expression of the cytokines. Moreover, MLD-STZ effects on the TGF-beta1 mRNA expression were reversed to the effects on IFN-gamma and TNF-alpha. The in vitro effects of STZ in islets, in general, were similar to those exerted by MLD-STZ. Apparently, reduction and upregulation of Th2-type cytokines was more associated with susceptibility and resistance, respectively, to MLD-STZ-induced diabetes than upregulation of Th1-type cytokine levels.

Animals↗

PubMed2002

Headache as an unusual presenting symptom of retropharyngeal ganglioneuroma.

Ganglioneuromas are rare benign neurogenous neoplasms. The clinical symptoms of ganglioneuromas of the neck are usually mild and non-specific and definitive diagnosis requires a histological examination. We present the case of a 35-year-old female who complained of retroauricular pain as her first symptom and who was initially diagnosed with migraine. ENT examination revealed a bulging of the left pharyngeal wall. Fine-needle aspiration biopsy was misleading in terms of diagnosis. Histology after extirpation showed a ganglioneuroma. Ganglion cell differentiation was proven using the new intermediate filament class alpha-internexin immunohistochemical staining technique. Headache is an uncharacteristic symptom of ganglioneuroma and an interdisciplinary approach is required to find a possible cause. Periauricular pain without abnormal otoscopic findings should lead the otorhinolaryngologist to consider a retropharyngeal condition, especially if combined with dysphagia. Complete resection of the tumor using modern microsurgical techniques is the best way to extract ganglioneuroma today.

Adult↗

PubMed2002

Assessments of corneal endothelial cell density in growing children and its relationship to horizontal corneal diameter.

BACKGROUND: Few studies have been published of corneal endothelial cell density (ECD) in children, especially in relation to the age and corneal size of the individuals. The possible effect of corneal enlargement on the endothelium was studied, and the results were objectively reviewed in relation to literature reports on corneal diameter and age. METHODS: ECD was assessed in 119 children aged 5 to 15 years after endothelial photography with a noncontact specular microscope (Topcon SP-1000 or SP-2000P), followed by manual digitization of cell overlays. The horizontal corneal diameter, as the visible iris diameter, was measured to 0.5-mm accuracy. RESULTS: The mean +/- SD value for ECD, calculated using all individual cell area values, was 3542 +/- 510 cells/mm2 (range, 2576 to 5316), whereas it was 3246 +/- 416 cells/mm2 (range, 2404 to 4817) if the mean cell area value from each image was used to calculate ECD. The mean horizontal corneal diameter was 11.48 +/- 0.58 mm (+/- SD). The corneal diameter was not obviously age-related for this group, but there were clear trends for younger children to have higher ECD values than older children (p2, depending on the method for calculating ECD. CONCLUSIONS: ECD values were found to be significantly correlated to corneal diameter in children aged 5 to 15 years, and this factor should be considered in comparative studies.

Adolescent↗

PubMed2002

Cooperative activation of human papillomavirus type 8 gene expression by the E2 protein and the cellular coactivator p300.

The E2 proteins of papillomaviruses (PV) bind to the coactivator CBP/p300 as do many other transcription factors, but the precise role of CBP/p300 in E2-specific functions is not yet understood. We show that the E2 protein of human PV type 8 (HPV8) directly binds to p300. Activation of HPV8 gene expression by low amounts of HPV8 E2 was stimulated up to sevenfold by coexpression of p300. The interaction between E2 and p300 may play a role in differentiation-dependent activation of PV gene expression, since we can show that the expression level of p300 increases during keratinocyte differentiation. Surprisingly, sequence-specific binding of E2 to its recognition sites within the regulatory region of HPV8 is not necessary for this cooperation, indicating that E2 can be recruited to the promoter via protein-protein interaction. HPV8 E2 binds via its N-terminal activation domain (AD), its C-terminal DNA binding domain (DBD), and its internal hinge region to p300 in vitro. Transient-transfection assays revealed that the AD is necessary and sufficient for cooperative activation with p300. However, we provide evidence that the interaction of the hinge and the DBD of HPV8 E2 with p300 may contribute. Our data suggest an important role of p300 in regulation of HPV8 gene expression and reveal a new mechanism by which E2 may be recruited to a promoter to activate transcription without sequence specific DNA binding.

Acetyltransferases↗

PubMed2002

Noncontact three-dimensional laser measuring device for tracheoscopy.

Luminal dimension measurements are still an unsolved problem in endoscopy. The goal of our study was to develop a 3-dimensional (3-D) measuring device for endoscopic applications. For this purpose, a fiber probe projecting a ring of laser light (laser diode, 675 nm) was integrated in a position registration system. Software was developed for image analysis, distortion correction, and export of data for 3-D display. Experimental evaluation of measuring accuracy employing plastic tubes and 15 postmortem pig trachea preparations with artificial stenosis indicated a high level of method precision (Pearson's correlation coefficients, r = 0.99 for normal tracheal lumen and r = 0.97 for high-grade stenosis; p < .0001). Our clinical experience with the technique in 10 patients with tracheal stenosis revealed no side effects and encourages us to recommend this method for other endoscopic applications. The method enhances our 3-D grasp of endoscopically examined lesions.

Animals↗

PubMed2002

Impact of vocal cord paralysis on cricoarytenoid joint.

To demonstrate structural changes in the cricoarytenoid joint after recurrent laryngeal nerve paralysis, we performed a laboratory investigation of fixed arytenoid cartilages from adult humans obtained during laser surgical arytenoidectomy in cases of bilateral vocal fold paralysis, analyzing the articular cartilage, the joint capsule, and the attached laryngeal musculature. Ten arytenoid cartilages from adult humans were studied by means of histology, as well as scanning and transmission electron microscopy. After long-standing denervation (>6 months), all arytenoid cartilages showed degenerative changes in their joint surface structure at various levels of intensity. The articular surface revealed fibrillation in some places, demasking of collagen fibrils next to the joint surface, and formation of chondrocyte clusters near the joint surface. All specimens also showed muscle atrophy. We conclude that long-standing recurrent laryngeal nerve paralysis does not result in ankylosis of the cricoarytenoid joint, as assumed, but the articular cartilage undergoes structural changes comparable to those in osteoarthritis. Structural changes in the articular cartilage and in the surrounding musculature hamper efforts at joint function recovery, as do procedures aiming solely at either medialization or lateralization of the vocal fold.

Adolescent↗

PubMed2000

Interaction with CBP/p300 enables the bovine papillomavirus type 1 E6 oncoprotein to downregulate CBP/p300-mediated transactivation by p53.

The E6 oncoprotein of bovine papillomavirus type 1 (BPV-1) can transform cells independently of p53 degradation. The precise mechanisms underlying this transformation are not yet completely understood. Here it is shown that BPV-1 E6 interacts with CBP/p300 in the same way as described for the E6 proteins of oncogenic human papillomaviruses. This interaction results in an inhibition of the transcriptional coactivator function of CBP/p300 required by p53 and probably by other transcription factors. The comparison of the CBP/p300-binding properties of BPV-1 E6 mutants previously characterized in transcription and transformation studies suggests (i) that the E6-CBP/p300 interaction may be necessary, but not sufficient, for cell transformation, and (ii) that the transcriptional activator function, inherent to the E6 protein, is not derived from forming a complex with CBP/p300.

Animals↗

PubMedDate not supplied

[Ultrasound biomicroscopy of conjunctival lesions].

BACKGROUND: The value of ultrasound biomicroscopy in the diagnosis of conjunctival lesions is not well established. PATIENTS AND METHODS: For the examination of conjunctival lesions, we used an ultrasound biomicroscope (Humphrey, Zeiss, Oberkochen) with a high frequency transducer (30 MHz). Between January 2000 and August 2001, 28 patients (16 female, 12-male) with conjunctival lesions, aged 9 to 81 years, were available for this study. RESULTS: Histological examination of the excised tissue displayed the presence of a compound naevus (8/28), cysts (6/28), inflammatory processes (3/28), granulomatous processes (2/28), lymphomas (2/28), foreign bodies (2/28), a pterygium (2/28), a malignant melanoma (1/28), a primary acquired melanosis (1/28), and a conjunctival amyloidosis (1/28). Using ultrasound biomicroscopy we were able to demonstrate a cystic tumour in the six patients (21 %) with a cyst of the conjunctiva. In patients suffering from solid tumours of the conjunctiva the definite diagnosis could not be made with ultrasound biomicroscopy alone. The eight patients with compound naevus displayed a somewhat heterogeneous sonographic structure within the tumour. In the patient with a foreign body we were able to demonstrate posterior shadowing of the underlying tissue. CONCLUSION: For evaluation of conjunctival lesions caused by a cyst or a solid tumour, ultrasound biomicroscopy may be an additional diagnostic tool, e. g. for assessing the margins of the tumour. However, up to now it is not possible to differentiate between different lesions solely by means of ultrasonography.

Adolescent↗

PubMedDate not supplied

Validation of primed in situ labeling for interphase analysis of chromosomes 18, X, and Y in uncultured amniocytes.

OBJECTIVE: Primed in situ labeling (PRINS) is an interesting alternative to the traditional fluorescence in situ hybridization (FISH) for the in situ detection of specific sequences in chromosome anomalies. It combines the sensitivity and specificity of the polymerase chain reaction with the specific in situ signal detection capability of FISH. METHODS: We performed PRINS on uncultured amniocytes and compared the results with standard cytogenetic analysis. In a prospective study, a total of 262 independent samples were analyzed for numerical aberrations of chromosome 18. RESULTS: In more than 95% of the cases PRINS reactions were successfully achieved. Neither false-positive nor false-negative results were obtained. 62 of the 262 cases were in parallel examined for chromosome 18 aneuploidies by FISH. Although there were significant differences in signal distribution, these did not lead to a different overall classification of the respective cases, i.e., the end results of disomic and trisomic cases for chromosome 18 were identical between FISH and PRINS. In 205 of 262 cases PRINS was performed for chromosomes X and Y. 97.6% of these samples were properly sex differentiated. CONCLUSIONS: The PRINS assay is a simple and cheap alternative to detect numerical aberrations of chromosome 18. However, the rate of false-positive results for chromosome X was calculated as 1% and the rate of false-negative ones for chromosome Y as 2%. Further investigations are required to transform PRINS into an alternative to conventional methods for routine rapid prenatal diagnosis of gonosomes.

Amnion↗