Biomedical subjects

M Hahn

Publications and source records attributed to M Hahn.

At least 37 records · Page 2Linked to original sources

PubMed2001

Influence of fluorophor dye labels on the migration behavior of polymerase chain reaction--amplified short tandem repeats during denaturing capillary electrophoresis.

The determination of the length of polymerase chain reaction (PCR)-amplified short tandem repeats (STRs) by denaturing capillary electrophoresis (CE) is a standard procedure for purposes of genotyping. We show that dye-specific mobility anomalies exist for 5'-fluorophor-labelled single-stranded DNA (ssDNA) fragments in CE using the performance-optimized polymer 4 (POP4) buffer sieving matrix, containing the entangled poly(N,N-dimethylacrylamide) polymer, urea, and 2-pyrrolidinone. The dye-specific retardation effects relative to coseparated GeneScan-500 [TAMRA] standard fragments can lead to wrong genotyping, even for allele-specific fragments of pentanucleotide STRs, when comparing the relative calculated sizes of identical fragments, labelled with rhodamine (ROX, TAMRA) or fluorescein dyes (FAM, 6-FAM, HEX, JOE, NED, TET): The size of fluorescein dye-labelled fragments of appr. 100 b in length appears to be smaller by up to 6.5 b. This effect becomes more dramatic with decreasing size: a 6-FAM-labelled 24-mer oligonucleotide appeared to be smaller by 11 b. In contrast, in classical urea/polyacrylamide slab-gel electrophoresis only a small dye-specific retardation of identical fragments is observed. The dye-specific effects are superimposed by weaker size and sequence-dependent anomalies of fragment mobility. Therefore, in denaturing CE the coseparation of a defined allele ladder labelled with the same dye as the unknown sample fragments remains the method of choice for accurate genotyping.

DNA, Single-Stranded↗

PubMed2001

[Status of hearing aid use by children in schools for the hearing impaired and deaf].

To evaluate and possibly improve the hearing aid fittings of children attending the Westphalian School for the Hearing Impaired or the Westphalian School for the Deaf, regular pedaudiologic consulting hours were established at both schools. During a 2-year period, 115 children were examined once, 35 children twice, and 5 children three times. The examinations comprised ear microscopy, audiometry, and a check of the hearing aids with a 0.6-cm3 coupler (children up to 7 years) or 2-cm3 coupler, respectively. The following criteria were used to assess the quality of the hearing aid setting: status of the external auditory canal and middle ear, acceptance of wearing the hearing aid, status of the ear mold, technical status of the hearing aid, and its setting. The results were related to four variables: gender, type of school, age, and mean hearing loss. Overall, just 40.9% of all children showed satisfactory hearing aid performance at the first examination and just 37.1% at the second. A significant influence of the variables on the hearing aid performance was documented for hearing loss only. The higher the hearing loss, the more likely the children were to have good hearing aid status. Analysis of the different parameters revealed that an incorrect setting was the main problem, with a rate of 20.9%; the rate of the other parameters varied from 6.1% to 15.7%. Thus, no parameter was of major relevance to the results. The results of the second examination were poorer in most parameters than those of the first. These alarming results, which are probably not only of regional significance, demonstrate that the hearing aid status of children attending schools for the hearing impaired or for the deaf is in urgent need of improvement.

Adolescent↗

PubMed2001

Structural analysis of high resolution in vitro MR images compared to stained grindings.

The recent advancement of high resolution magnetic resonance imaging has opened up new avenues for the determination of structural characteristics of the trabecular network, which may significantly improve the diagnosis of osteoporosis. An analysis of the calcaneus in healthy women has shown similar age-related changes when comparing structural parameters in high resolution MR images and BMD as measured by DXA [1]. Here we undertook an in vitro study to further compare structural measurements in MR images with those from stained grindings. A 3D gradient echo sequence on a 1.5 T scanner was used to obtain four contiguous sagittal MR images with a slice thickness of 1 mm and an in plane pixel size of 195 microns. Twenty-one stained grindings with a slice thickness of 1 micron each were obtained from a 3 mm thick slab of the same volume investigated by MR. The stack of stained grindings was also used to simulate the influence of variations in slice thickness and in plane resolution. Results for structural parameters derived from the high resolution MR images differed considerably from those derived from the stained grindings because the MR images are heavily influenced by partial volume artifacts. This finding was supported by simulations which also revealed that even at a slice thickness of 500 microns and an in plane pixel size of 13 microns, accurate results could not be obtained when a histomorphometric type analysis was applied. Results also depended strongly on the segmentation method. However, contrary to the stained grindings, images averaged over several slices reveal the three-dimensional network character of the trabecular structure. New efforts should be undertaken to develop analysis strategies that are more adequate for in vivo high resolution images instead of using analysis techniques applied in classical histomorphometry.

Cadaver↗

PubMed2001

Interaction of cisplatin with methionine- and histidine-containing peptides: competition between backbone binding, macrochelation and peptide cleavage.

The pH- and time-dependent reaction of cis-[PtCl2(NH3)2] with the methionine- and histidine-containing peptides H-Gly-Met-OH, H-Gly-Gly-Met-OH, Ac-His-Gly-Met-OH, and Ac-His-(Ala)3-Met-OH at 313 K has been investigated by ion-pairing reverse phase HPLC and NMR spectroscopy. For equimolar solutions (c=0.8 mM, pH approximately equals 3 or 8.8), initial formation of the kinetically favored S-bound complex is followed by relatively rapid metallation of the neighboring methionine amide nitrogen NM to afford a kappa2NM,S six-membered chelate. The strong trans effect of the methionine S then favors facile NH3 substitution, leading to generation of tridentate complexes such as [Pt(H-Gly-MetH(-1)-OH)-kappa3NG,NM,S)(NH3)]+ or [Pt(H-Ac-His-GlyH(-1)-MetH(-1)-OH-kappa3NG,NM,S)(NH3)]. In the case of H-Gly-Gly-Met-OH, this reaction is accompanied by loss of a second NH3 ligand in alkaline solution to generate the tetradentate kappa4NG1,NG2,NM,S species. In contrast, cleavage of the backbone C(O)-N bond to the second metallated amide nitrogen after t>100 h is common to the tridentate complexes of the tri- and pentapeptides at pH<5. Although an imidazole-coordinated kappa2N3H,S macrochelate is formed throughout the whole range 2.5 < or = pH < or = 10 for Ac-His-Gly-Met-OH, it slowly decays (t=10-1000 h) to the thermodynamically more stable tridentate kappa3NG,NM,S complex. All major final products were separated and fully characterized by NMR and MS.

Antineoplastic Agents↗

PubMed2001

Inter-laboratory comparison of DNA preservation in archival paraffin-embedded human brain tissue from participating centres on four continents.

DNA extracted from formalin-fixed and paraffin-embedded brain tissue is known to contain as yet ill-characterized inhibitors of the PCR process. As part of a project that aims to clarify the role of mitochondrial DNA sequence variation in human neurodegenerative diseases using DNA from various ethnic backgrounds, we have investigated factors that influence the preservation of archival DNA and its suitability for PCR. In this study, neuropathological tissue samples were analysed that had been routinely processed in 18 international centres on four continents. Following DNA extraction, PCR amplification of mitochondrial and nuclear DNA sequences was performed with and without additional purification of the template DNA. In addition, the DNA used for PCR was analysed by HPLC. Phosphate-buffered formalin proved to be a superior fixative compared with unbuffered aldehyde: DNA extraction resulted in greater yields, the molecular weight of the isolated DNA was higher and PCR was more successful. PCR inhibitors were identified as (1) high concentrations of small (<300 bp) DNA fragments that competitively compete with template DNA and (2) contaminants of the DNA template solution including denatured protein that cannot be completely removed by phenolic extraction. HPLC analysis did not reveal significant qualitative differences between DNA isolated from fresh-frozen tissue samples and DNA recovered from formalin-fixed, paraffin-embedded brain tissue. The fact that DNA could be amplified from the majority of tissue specimens in this study suggests that rare diseases and diseases where ethnic background plays an important role can be sampled for genetic polymorphism analysis on a global scale using archival neuropathological collections.

Alzheimer Disease↗

PubMed2001

The Structure of calnexin, an ER chaperone involved in quality control of protein folding.

The three-dimensional structure of the lumenal domain of the lectin-like chaperone calnexin determined to 2.9 A resolution reveals an extended 140 A arm inserted into a beta sandwich structure characteristic of legume lectins. The arm is composed of tandem repeats of two proline-rich sequence motifs which interact with one another in a head-to-tail fashion. Identification of the ligand binding site establishes calnexin as a monovalent lectin, providing insight into the mechanism by which the calnexin family of chaperones interacts with monoglucosylated glycoproteins.

Amino Acid Motifs↗

PubMed2001

Signal and nutrient exchange at biotrophic plant-fungus interfaces.

Biotrophic interfaces are formed in mutualistic and parasitic plant-fungus interactions. They result from coordinated developmental programs in both partners and represent specialized platforms for the exchange of information and nutritional metabolites. New data on the establishment and the components of functional interfaces have been obtained in a number of ways. First, by isolation of symbiotically defective mutants; second, by characterization of new genes and their products; and, third, by the identification and localization of components of biotrophic interfaces, such as cell-wall proteins, H+-ATPases and nutrient transporters.

Adenosine Triphosphatases↗

PubMed2001

Estimating the relapse stage in chronic myeloid leukaemia patients after allogeneic stem cell transplantation by the amount of BCR-ABL fusion transcripts detected using a new real-time polymerase chain reaction method.

We have used a new single-step real-time reverse transcription polymerase chain reaction (RT-PCR) method to quantify BCR-ABL transcripts, thereby estimating the relapse stage in chronic myeloid leukaemia patients after allogeneic transplants. In 402 samples from 172 patients, BCR-ABL expression was determined and normalized, using the GAPDH housekeeping gene product as an endogenous reference. In our real-time RT-PCR assay, serial dilutions of RNA of the K562 cell line remained positive down to 7.5 pg. The median normalized BCR-ABL amount differed significantly (P < 0.001) between the various disease stages and was 0.06% (range 0.001-1.55%), 3.2% (range 1.4-5.6%) and 21.5% (range 6.8 -827%) in 17 patients with a molecular relapse, in eight patients with a cytogenetic relapse and in 10 patients with a haematological relapse respectively.

Adolescent↗

PubMed2001

Chromoendoscopy with indigocarmine improves the detection of adenomatous and nonadenomatous lesions in the colon.

BACKGROUND AND STUDY AIMS: Depressed early cancers and flat adenomas have a high potential for malignancy with possible infiltrating growth, despite the small size of the lesion. Japanese investigators have shown that early diagnosis and classification of these lesions is possible with the help of chromoendoscopy. The aim of this study, therefore, was to evaluate the usefulness of chromoendoscopy during routine colonoscopy. PATIENTS AND METHODS: During routine colonoscopy, vital staining with indigocarmine solution (0.4 %, 1 - 10 ml) was performed on all visible lesions in 100 consecutive patients without visible inflammatory changes. If findings on macroscopic examination were unremarkable, the sigmoid colon and rectum were stained with indigocarmine over a defined segment (0 - 30 cm ab ano) and inspected for lesions visible only after staining. Each lesion was classified with regard to type (polypoid, flat, or depressed), position and size. The staining pattern was classified according to the pit pattern classification. RESULTS: A total of 52 patients had 105 visible lesions (89 polypoid, 14 flat and two depressed). The mean size of the lesions was 1.4 cm. Among the 48 patients with mucosa of normal appearance, 27 showed 178 lesions after staining (176 flat, two depressed) with a mean size of 3 mm. On histological investigation, 210 lesions showed hyperplastic or inflammatory changes, 67 were adenomas and six were cancers. Use of the pit pattern system to classify lesions (adenomatous, pit patterns III-V; nonadenomatous, pit patterns I-II) was possible, with a sensitivity of 92 % and a specificity of 93 %. Lesions with pit patterns III - V showed higher rates of dysplasia. CONCLUSIONS: Chromoendoscopy allows easy detection of mucosal lesions in the colon and facilitates visualization of the margins of flat lesions. This technique unmasks multiple mucosal lesions which are not identified by routine video colonoscopy. The pit pattern seen after staining allows differentiation between hyperplastic and adenomatous lesions which may have consequences with regard to the endoscopic interventions needed.

Adenoma↗

PubMed2001

[Interventional ERCP in patients with cholestasis. Degree of biliary bacterial colonization and antibiotic resistance].

Interventional ERCP in patients with cholestasis. Degree of biliary bacterial colonization and antibiotic resistance. Biliary obstruction together with bacterial colonization of the bile duct may lead to development of acute cholangitis. The aim of our prospective study was to investigate the presence and degree of biliary bacterial colonization by means of bile aspiration during ERCP in patients with biliary obstruction. Furthermore, we evaluated antibiotic therapy regimens, which would cover the bacterial species obtained by ERCP and subsequent culture in each patient. In addition, analysis of risk factors was performed that would predispose to the development of cholangitis.80 patients with clinical and laboratory evidence of biliary obstruction underwent ERCP with initial aspiration of bile via the cannulation catheter. This material was used to culture aerobic and anaerobic bacteria and determine the colony count/ml bile, followed by identification of each species and antibiotic resistance testing. The minimal inhibitory concentration for Levofloxacin, Ciprofloxacin, Piperacillin, Ampicillin, Ceftriaxone, Imipenem, Gentamycin und Metronidazole was determined. Immediately after the ERCP or if the body temperature (after ERCP) rose to > 38 degrees C blood cultures were obtained. In 45 (56 %) patients biliary colonization with bacteria could be identified (56 %). In 20 patients a single isolate was cultured, in 25 cases mixed infection was present. A total of 83 species were isolated. The most common bacteria were E. coli, Enterococcus and Klebsiella. 9.6 % of all isolates were obligatory anaerobes. In 9 of 80 patients bloodcultures tested positive for bacterial growth (rate of bacteremia: 11.3 %). 10 patients had acute cholangitis clinically before ERCP, 13 patients developed signs of infection after ERCP. Statistically significant factors contributing to the risk of infection were age of the patient, the clinical condition of the patient before ERCP and the biliary colony count. Patients with development of infection after ERCP showed a significantly higher incidence of bacterial colonization of the biliary tree and a higher colony count. In all bacterial species Imipenem (4.5 %) or Levofloxacin (2.2 %) exhibited the lowest rate of in-vitro resistance. Based on these data, the implementation of Levofloxacin in combination with anaerobic coverage is advantageous as a calculated therapy for patients with acute cholangitis.

Adult↗

PubMed2001

Screening for specialized columnar epithelium with methylene blue: chromoendoscopy in patients with Barrett's esophagus and a normal control group.

BACKGROUND: Barrett's esophagus is characterized by the presence of specialized columnar epithelium (SCE) in the distal esophagus and is a precursor for dysplasia and adenocarcinoma. Methylene blue is used to selectively stain this type of epithelium within the columnar lined distal esophagus (CLE). The goal of our prospective study was to investigate the staining pattern with methylene blue in patients with visible CLE and a control group with macroscopically normal gastroesophageal junction. METHODS: Chromoendoscopy of the distal esophagus with 1% methylene blue was performed on a total of 73 patients (51 with visible CLE, 22 normal control subjects). Biopsies were targeted toward stained or macroscopically abnormal mucosal areas. Unstained CLE was sampled by obtaining 4-quadrant biopsy specimens at 2 cm intervals. RESULTS: The targeted biopsy of stained areas provided histologic proof of SCE with a sensitivity of 98% and a specificity of 61%. The detection of SCE increased with this method in patients with visible CLE (75%), but SCE was also prevalent in the control group (73%). Areas that by their appearance raised a suspicion for dysplasia and adenocarcinoma were already endoscopically apparent in 3 of 4 cases before methylene blue staining and showed different staining patterns. CONCLUSIONS: Methylene blue staining increases the rate of detection of SCE, both in patients with visible CLE and with normal gastroesophageal junction. For detection of dysplastic or malignant areas it is important to look for visible mucosal changes.

Aged↗

PubMed2001

Frequency of mediastinal lymph node metastases in patients undergoing EUS evaluation of pancreaticobiliary masses.

BACKGROUND: Mediastinal lymph node metastases have rarely been reported in patients with pancreatic cancer. Our aim was to determine the frequency of mediastinal lymph node metastases in patients with pancreaticobiliary masses by using EUS-guided fine needle aspiration. METHODS: Sixty-six consecutive patients with pancreatobiliary masses were evaluated on EUS for the presence of mediastinal lymph node metastases. All masses were staged by commonly used EUS criteria by using sector scanning echoendoscopes. Mediastinal lymph nodes with EUS features that suggested malignancy were aspirated. RESULTS: Of the 66 patients (mean age 65.6 years; 38 men), 4 had biliary masses, 5 had lesions of the major duodenal papilla, and 57 had pancreatic masses. Eleven patients (10 pancreatic masses, 1 biliary mass) had enlarged mediastinal lymph node (12-30 mm) on EUS; in 2 patients these had a benign appearance and were not aspirated. Nine patients underwent EUS-guided fine needle aspiration: in 1 the cytology was inconclusive (patient subsequently had a negative Whipple resection); in 4 the mediastinal lymph node cytology was benign; the remaining 4 patients had adenocarcinoma cells in the aspirate from mediastinal lymph node. These 4 pancreatic tumors were staged by EUS as T2N1M1 (1), as T4N0M1 (2, one later found to also have a lung mass), and T4N1M1 (1). CONCLUSION: Enlarged mediastinal lymph nodes were found on EUS in 16.6% (95% CI [7.7%, 25.6%]) of patients with pancreatobiliary masses and in 17.5% (95% CI [7.6%, 27.4%]) of patients with pancreatic masses. The frequency of mediastinal lymph node metastases in pancreatobiliary masses was 6.1% (95% CI [0.34%, 11.9%]) and in pancreatic masses 7.0% (95% CI [0.4%, 13.6%]). Routine EUS evaluation of the mediastinum in patients with pancreatic masses is warranted.

Adenocarcinoma↗

PubMed2001

Three-dimensional visualization of coronary arteries in excised hearts.

OBJECTIVE: We sought to image coronary arteries in excised hearts. METHODS: Twelve excised pigs' hearts were imaged in a water bath. The aortic valve was closed surgically. A contrast agent (Echovist) was injected into the aortic root and selectively into single coronary arteries. Three-dimensional (3D) imaging was performed with TomTec Echoscan equipment. Mechanical rotations were performed at 1 degrees intervals. The hearts were visualized by InVivo software. Selective coloring of coronary arteries in 3D data sets was obtained by using color superpositioning, which differentiates information before and after injection of contrast. Distance measurements were performed in conventional 3D echocardiograms of coronary arteries and color-superimposed echocardiograms and compared with those from angiograms and casts. RESULTS: After a learning curve, during which optimal conditions for the visualization of coronary arteries were determined, a quick display of all major parts of the coronary tree was obtained. Distance measurements (n >400) revealed that fundamental contrast echocardiography overestimated angiography by 25% +/- 5% and casts by 28% +/- 6%. However, distances in color-superimposed echocardiograms (flow mode 4) were not significantly different from those obtained from angiograms and casts. In harmonic contrast echocardiograms, color super-positioning gave smaller distances compared with those from fundamental contrast echocardiograms, though they were still significantly larger than the reference diameters. CONCLUSIONS: The 3D imaging of epicardial coronary arteries under ideal conditions in a water bath seems feasible and provides insight into coronary visualization with the use of ultrasonography.

Animals↗

PubMed2001

The influence of hypothermia on outer hair cells of the cochlea and its efferents.

Transient evoked otoacoustic emissions (TEOAE) were recorded in 21 guinea-pigs undergoing hypothermia. The minimal average body temperature during cooling was 26 degrees C/24.9 degrees C measured orally or rectally, respectively. The animals were subsequently warmed to normal body temperature. A clear influence of body temperature on TEOAE could be documented. During cooling the amplitude and reproducibilities decreased, disappearing completely at a mean temperature below 28.5 degrees C (oral) and 27.3 degrees C (rectal). The emissions reappeared during rewarming at a mean temperature of 30.1 degrees C (oral) and 30.8 degrees C (rectal). Contralateral auditory stimulation (CAS) led to a decrease of the amplitudes of TEOAE during cooling down to a mean of 33 degrees C/32 degrees C (oral/rectal temperature). During rewarming, influences of the CAS could be recognized, again at an oral temperature above 35 degrees C. The changes to the TEOAE observed in these experiments suggest that hypothermia affects not only the outer hair cells (OHC) of the cochlea but also the efferent supply to the cochlea.

Acoustic Stimulation↗

PubMed2001

Comparison between Taq DNA polymerase and its Stoffel fragment for quantitative real-time PCR with hybridization probes.

In quantitative real-time PCR assays, fluorophor-labeled oligonucleotide probes are employed to generate sequence-specific signals for the quantitative evaluation. Whereas TaqMan probes have to be hydrolyzed during PCR by the endonucleolytic activity of Taq DNA polymerase to generate a signal, the hybridization probes in LightCycler assays must not be hydrolyzed. In this study, we demonstrate for four different targets that the probes are degraded during PCR by Taq DNA polymerase. Signal yield, quality of amplification curves, and accuracy of quantitative measurements can be improved using the Stoffel fragment lacking an endonucleolytic activity and TaqStart antibody suppressing the formation of nonspecific products, without laborious efforts to optimize the amplification protocol.

DNA-Directed DNA Polymerase↗

PubMed2001

Venous leg ulcers and microcirculation.

Incompetence of venous valves do not lead per se to trophic skin changes of the lower limb. In severely concerned patients suffering from dermatoliposclerosis or venous leg ulcers retrograde pressure waves were detected in the nutritive capillaries of the skin by the "servo nulling" pressure measurement during simulated calf muscle contraction. Between the degree of trophic skin change and the capillary density and cutaneous oxygen partial pressure a close correlation was found. Healing of venous ulcers occurred only if the cutaneous microangiopathy in the ulcer area improved. Capillary density and transcutaneous oxygen partial pressure were of prognostic impact as to healing of venous ulcers.

Blood Pressure↗

PubMed2001

Time independence of the prognostic impact of tumor cell detection in the bone marrow of primary breast cancer patients.

PURPOSE: Tumor cell detection (TCD) in bone marrow is an outstanding prognostic factor in breast cancer. There is only one other study that has investigated more than 300 patients with a median follow-up of more than 5 years (J. L. Mansi et al., Lancet, 354:197-202, 1999). We report data from 727 patients with a median follow-up period of 6.5 years. EXPERIMENTAL DESIGN: In a prospective study, intraoperatively aspirated bone marrow was screened for micrometastatic cancer cells. We used an immunocytological method (monoclonal mucin antibody 2E11; the avidin-biotin complex method). RESULTS: Forty-three percent of the patients were TCD positive. Sixty percent of the patients with distant metastases were tumor cell positive (155 of 258 patients). Forty-nine percent of the patients with positive TCD developed distant metastases (155 of 315 patients). TCD was an independent prognostic factor for clinical outcome after a median follow-up time of 6.5 years. The prognostic impact of TCD and tumor size remains constant with the time, whereas the impact of grading and progesterone receptor on risk seems to decrease with longer follow-up time. CONCLUSIONS: TCD remains an independent prognostic factor The impact of TCD does not change with longer follow-up time. TCD is a reliable prognostic factor and provides important information about the process of metastasis.

Adult↗

PubMed2000

Enhancing effect of polyoxyethylene alkyl ethers on the skin permeation of ibuprofen.

The influence of polyethoxylated non-ionic surfactants on the transport of ibuprofen across rat skin was investigated. The skin permeation of ibuprofen from a series of 17 polyoxyethylene (POE) alkyl ethers containing 5% ibuprofen was determined using Franz diffusion cells fitted with excised rat skins. Differential scanning calorimetry (DSC) and Fourier transform infrared spectroscopy (FT-IR) were performed for the physicochemical characterization of ibuprofen-surfactant interaction. In vitro transdermal flux through excised rat skin was found in the decreasing order of POE(5)cetyl/oleyl ether (110.24 microg/cm(2)/h)>POE(2)lauryl ether (99.91 microg/cm(2)/h)>POE(2)oleyl ether (67.46 microg/cm(2)/h)>POE(10)stearyl ether (66.19 microg/cm(2)/h). POE(2)oleyl ether showed the longest lag time (2.47 h). The enhancers containing the EO chain length of 2-5, HLB value of 7-9 and an alkyl chain length of C16-C18 were effective promoters of ibuprofen flux. FT-IR and DSC studies to probe the nature of the interaction between the ibuprofen and surfactant indicated that the hydrogen bonding state of ibuprofen was changed from the dimeric form to the carbonyl-hydroxyl (C=O-HO) hydrogen bond form in the presence of excess POE alkyl ether. These results indicated that this new system may be used in developing a transdermal formulation with improved skin permeation of ibuprofen.

Algorithms↗