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M Hahn

Publications and source records attributed to M Hahn.

At least 73 records · Page 4Linked to original sources
PubMed2000

Influence of DNA target melting behavior on real-time PCR quantification.

BACKGROUND: Quantitative real-time PCR is increasingly used to quantify copy numbers of nucleic acids for clinical applications. We observed that the measurements of allele imbalances of the tumor suppressor gene p16 and the oncogene ErbB-2 yielded results with variable precision under certain experimental conditions. METHODS: We used the LightCycler(TM) real-time PCR system to quantify different genomic target sequences using hybridization probes or SYBR Green for detection. RESULTS: With two primer/template systems (p16 and ErbB-2), we observed sinusoidal scattering of the threshold cycle values depending on the capillary position in the thermostated reaction chamber. This scattering depended on the denaturation temperature only when complete genomic DNA was used as template and did not occur when PCR product or restricted or boiled genomic DNA was used or the denaturation temperature in the first cycles was increased (and other targets, such as p53, HBB, IGF-1, GAPDH, and PBGD, did not show this behavior). CONCLUSIONS: Before a primer system is used for precise quantitative real-time PCR, the dependence of the quantification results on the positions of reaction tubes in the thermocycler should be tested. Our data indicate that amplification efficiencies, especially in the first cycles, depend not only on the priming efficiencies of the primers and the melting temperature of the amplicon, but also on the melting behavior of the amplicon's genomic vicinity. Complete denaturation of genomic DNA is necessary to maximize precision of quantitative PCR. Higher denaturation temperatures in the initial cycles or boiling of DNA before the PCR can improve the accuracy of quantification in some cases.

Cyclin-Dependent Kinase Inhibitor p16↗
PubMed2000

Microcirculatory dysfunction in chronic venous insufficiency (CVI).

The elevated ambulatory pressure in the peripheral venous system of chronic venous insufficiency (CVI) patients manifests itself not only in the form of disturbed macrocirculation but also and particularly in microangiopathic changes. For this reason, it is closely correlated with trophic disorders of the skin and can ultimately lead to ulceration. Using microcirculation research techniques, we are able to provide clear evidence of a typical microangiopathy in chronic venous insufficiency. Fifty CVI in Widmer stages I, II, and III were examined with fluorescence video microscopy, intravital video capillaroscopy, transcutaneous oxygen partial pressure measurement, TcpO2 and laser Doppler flowmetry. The effects of compression therapy with individually fitted compression stockings on capillary morphology were studied over a period of 4 weeks in 20 CVI patients in Widmer stages I and II. The capillary pressure was measured during simulated muscle contraction using a servo-null micropressure system. We periodically drew blood from the dorsalis pedis vein and a brachial vein of 11 healthy test persons and 8 patients with stage III CVI during experimental venous hypertension in order to evaluate the expression pattern of leukocyte adhesion molecules involved in inflammation: LFA-1 (CD11a), Mac-1 (CD11b), p150,95 (CD11c), CD18, VLA-4 (CD49d), and L-selectin (CD62L). In the same patients, we used immunohistochemical methods to examine clinically unaffected skin and the skin near an ulcer, focusing on the adhesion molecules ICAM-1, VCAM-1, and E-selectin. The microangiopathic changes observed with worsening clinical symptoms include a decrease in the number of capillaries, glomerulus-like changes in capillary morphology, a drop in the oxygen content (tcpO2) of the skin, increased permeability of the capillaries to low-molecular-weight substances, increased laser Doppler flux reflecting elevated subcutaneous flow, and diminished vascular reserve. These microangiopathic changes worsen in linear proportion to the clinical severity of chronic venous insufficiency. In patients with venous ulcerations, the baseline expression of LFA-1 and VLA-4 on lymphocytes, Mac-1 expression on the myeloid cell line, and L-selectin expression on all three cell lines was not significantly different form that in healthy controls. During orthostatic stress, there was a significant reduction in the expression of L-selectin in blood cells collected at foot level in the controls (p=0.002), but not in the patients. Clinical improvement by compression therapy was accompanied by an increase in the number of nutritive capillaries, while the diameter of the capillaries and the dermal papillae was reduced. When ulcers healed in a short period (<6 weeks), we observed a concomitant increase in the number of capillaries (p<0.05). Microangiopathy appears before tropic disorders of the skin develop. Even trophically normal skin areas may have dilated nutritive capillaries, an early sign of disturbed skin perfusion. These changes represent a plausible explanation for the development and to recurrency tendency of venous ulcers. The reduced expression of lymphocytic L-selectin in healthy controls during the orthostatic stress test may be an indication that the cells are activated by venous stasis. Clinically effective therapeutic measures improve the impaired microcirculation of the skin in the ankle area.

Bandages↗
PubMed1999

Hearing screening in healthy newborns: feasibility of different methods with regard to test time.

Clicked-evoked otoacoustic emissions and automated auditory brainstem response are both considered appropriate hearing screening tools. The aim of this study was to compare their practicability with respect to test duration. In the first part of the study, clicked-evoked otoacoustic emissions were recorded in 388 healthy infants on the newborn ward in the first days of life. To reduce false positive rates of screening, clicked-evoked otoacoustic emissions combined with automated auditory brainstem response measurement (A-ABR, Algo 1E and Algo 2) were used in a second investigation. Fifty-five newborns screened by this two-stage method recorded an increased pass rate. The new digital technique of automated auditory brainstem response measurement (Algo 1E and Algo 2) employed in the second part of the study has a test time even shorter than that of clicked-evoked otoacoustic emissions and is no longer a time-consuming test due to improved technology of the second generation of automated auditory brainstem response equipment. Screening including preparation and follow-up time took 9.43 min (Algo 1E) and 8.26 min (Algo 2) respectively and was thus less time-consuming than reported in former studies.

Audiometry, Evoked Response↗
PubMed1999

Local cold exposure test with a new arterial photoplethysmographic sensor in healthy controls and patients with secondary Raynaud's phenomenon.

Local cold exposure tests are used to diagnose cold-induced vasospastic disorders and to evaluate therapeutic success. We investigated the pulsatile signal detected with a newly developed arterial photoplethysmography (APPG) method and the signal change induced by local cold exposure using a temperature-controlled finger holder, comparing it with laser Doppler flux (red and green laser, rLDF and gLDF) and red blood cell velocity measured in nailfold capillaries (CBV). Ten healthy volunteers and 10 age- and sex-matched patients suffering from Raynaud's phenomenon due to systemic sclerosis were investigated using a moderate cooling temperature of 16 degrees C for 5 min. All signals were recorded simultaneously. The results show a significant reduction of CBV (P < 0.0001), rLDF (P < 0.0003), and gLDF (P = 0.0214) during cooling and characteristic changes in the APPG signal (for instance a decrease in pulse wave amplitude; P < 0.0001). Significant differences in the APPG amplitude could be detected under resting conditions; at cooling temperatures there were also significant differences in CBV and APPG. The temperature-controlled finger holder with its built-in APPG probe appears to be a useful tool for evaluating the effect of local cooling on finger skin perfusion and differentiating between healthy controls and patients with secondary Raynaud's phenomenon due to systemic sclerosis. The gLDF signal was rather weak, limiting its value in cold stress tests. The differences between controls and patients in CBV were somewhat smaller than in previous studies, suggesting the advantage of lower local cooling temperatures, e.g., 12 degrees C.

Adult↗
PubMed1999

[Preventive surgery of inherited colorectal cancer identified through molecular diagnosis].

Numerous inherited genetic changes predisposing to cancer have already been identified and the number is increasing. Accurate prediction of individual risk by means of molecular diagnosis implies clinical consequences in the treatment of cancer-predisposing syndromes. Using familial adenomatous polyposis (FAP) and hereditary non-polyposis colorectal cancer (HNPCC) as an example, we present here the underlying genetic changes that contribute to tumor development. These genetic alterations can be efficiently identified through molecular diagnostic techniques. Identification of the familial germline mutation permits one to distinguish mutation carriers from non-mutation carriers within affected families and results in individually tailored surveillance and prevention. Therefore, molecular diagnosis is making a contribution to the advances in preventive surgical therapy. The indications are discussed.

Colorectal Neoplasms↗
PubMed1999

Hereditary colorectal cancer: clinical consequences of predictive molecular testing.

The continuing increase in knowledge about the genetic basis of carcinogenesis has led to diverse efforts to exploit this knowledge clinically, primarily in the form of predictive genetic testing. In conjunction with family history, gene tests are intended to improve individual cancer risk assessment. The objectives of predictive molecular testing are to identify the disease-causing germline mutation in an index person who has already developed the disease and to distinguish asymptomatic mutation carriers from non-mutation carriers within a given cancer-prone family. At present, genetic testing for colorectal cancer risk, primarily in form of DNA sequencing, is applied in familial adenomatous polyposis (FAP) and hereditary nonpolyposis colorectal cancer (HNPCC). In these inherited colorectal tumor syndromes determining the genetic status may result in an individually tailored surveillance program and prophylactic treatment. The implications of genetic testing for the clinical management of disease, both of mutation and non-mutation carriers, in familial adenomatous polyposis and hereditary nonpolyposis colorectal cancer families are discussed.

Adenomatous Polyposis Coli↗
PubMed1999

Collagen type I metabolism after bone surgery.

This study follows the postoperative course of serum collagen type I metabolites in patients after uncomplicated implantation of a cemented total hip endoprothesis (TEP; n = 12, mean age: 69.3 years), a cemented hemiendoprothesis (HEP; n = 13, mean age 79.7 years), a dynamic condylar or hip screw (DCS/DHS; n = 12, mean age 75.1 years) and osteosynthetic treatment of a Weber B or C fracture (OS; n = 17, mean age 54.3 years). The course of the propeptide of human type I procollagen (PICP) as an anabolic marker as well as of I-carboxyterminal telopeptide (ICTP) as a catabolic marker of bone metabolism was characterized. Measurements were done preoperatively and weekly for 3 weeks after surgery. The concentrations of both markers increased and reached a maximum in the 2nd or 3rd week after surgery. However, the PICP values differed, depending on the kind of surgical intervention and the type of bone healing. Secondary fracture healing with formation of callus occurred in the DCS/DHS group, which developed the highest median PICP concentrations (initial 83 microg/l, second week 337 microg/l; P < 0.001). In contrast, the primary bone healing in the OS group showed increasing ICTP but unchanged PICP concentrations. Patients in the cemented TEP and HEP groups as a kind of artificial bone healing had comparable concentrations. To consider the effective metabolism of collagen type I, the PICP/ICTP ratio was calculated. Although the median PICP and ICTP concentrations of the studied groups differed, the PICP/ICTP ratios were similar. In comparison to 54 young and healthy volunteers (median PICP/ICTP ratio: 37), the ratios of the studied groups were still normal but low (median ratios: < 20). This could be an effect of decreasing collagen type I metabolism with age. Although the results are in agreement with animal studies and histomorphometric investigations, the clinical use of PICP and ICTP determination as a tool for the detection of complicated bone healing is limited by the marked interindividual variability and the uncertain bone specificity.

Aged↗
PubMed1999

[Pt(dien)]2+ migrates intramolecularly from methionine S to imidazole Nepsilon2 in the peptides H-His-Gly-Met-OH and Ac-His-Ala-Ala-Ala-Met-NHPh.

The pH- and time-dependent reaction of [Pt(dien)(H2O)]2+ with the methionine- and histidine-containing peptides H-His-Gly-Met-OH and Ac-His-Ala-Ala-Ala-Met-NHPh at 313 K has been investigated by HPLC and NMR spectroscopy. For both peptides, initial relatively rapid formation of the kinetically favoured methionine S-bound complex is followed by slow intramolecular migration of the [Pt(dien)]2+ fragment to imidazole Nepsilon2 (or, in the case of H-His-Gly-Met-OH, to a much lesser extent to the competing imidazole Ndelta1) of the histidine side chain over a period of 500 h. Time-dependent studies for the pentapeptide at pH 8.0 demonstrate that this isomerization can take place by either direct S-->Nepsilon2 migration or by a two-step mechanism involving initial Nepsilon2 coordination of a second [Pt(dien)]2+ fragment and subsequent cleavage of the orginal Pt-S bond in the resulting dinuclear complex. The rate of kappaS/kappaNepsilon2 isomerization is markedly reduced on lowering the pH to 5.1.

Amino Acid Sequence↗
PubMed1999

A randomized trial of peroral versus transnasal unsedated endoscopy using an ultrathin videoendoscope.

BACKGROUND: Potential advantages of unsedated endoscopy include the prevention of side effects or morbidity related to the use of sedative drugs, less intensive patient monitoring, and less expense. We compared transnasal (T-EGD) with peroral (P-EGD) unsedated endoscopy by using an ultrathin video instrument with respect to patient tolerance and acceptance. METHOD: Patients were randomized to T-EGD or P-EGD. If the initial route of insertion failed, the patient was crossed over to the other route. If this also failed, the patient underwent endoscopy under conscious sedation with an ultrathin instrument. A questionnaire for tolerance was completed by the patient (a validated 0-10 scale where "0" represents none/well tolerated and "10" represents severe/poorly tolerated). RESULTS: Of 105 recruited patients, 60 consented to undergo unsedated endoscopy. There were 20 men and 11 women (mean age 45 years) in the P-EGD group and 15 men and 14 women (mean age 48 years) in the T-EGD group. Of 35 total P-EGD patients (4 were crossed over T-EGD patients), 34 (97%) completed an unsedated examination. Of 29 T-EGD patients, 25 (86%) had a complete examination. Three T-EGD examinations failed for anatomical reasons; all 3 patients when crossed over to the P-EGD route had a successful examination. One patient was unable to tolerate either route. Between the P-EGD and the T-EGD groups, pre-procedure anxiety (3.6 +/- 0.5 vs. 3.0 +/- 0.6), discomfort during insertion (2.1 +/- 0.5 vs 3.3 +/- 0.7), gagging (4.7 +/- 0.5 vs. 3.2 +/- 0. 6), and overall tolerance (2.4 +/- 0.5 vs. 3.8 +/- 0.7) were similar (p > 0.05). However, discomfort on insertion was significantly greater in the T-EGD versus the P-EGD group (4.4 +/- 0.6 vs. 2.7 +/- 0.5: p < 0. 05). Eighty-nine percent of P-EGD patients and 69% of T-EGD patients, p = 0.07, were willing to undergo unsedated endoscopy in the future. CONCLUSION: T-EGD patients experienced significantly more pain on insertion than did P-EGD patients. Otherwise, unsedated endoscopy by either the transnasal or the peroral route is generally well tolerated. In this study it was completed in 59 of 60 patients.

Adult↗
PubMed1999

Nonlinear analysis of blood flux in human vessels.

Laser Doppler fluxmetry (LDF) is frequently used in research on microcirculation of blood. Usually LDF time series are analysed by conventional linear methods, mainly Fourier analysis. These methods may not be optimal for the investigation of nonlinear effects of vasomotion, heartbeat or vessels. Nonlinear methods are based on a reconstruction of the system trajectory in an embedding space describing not only the measured time series but the behaviour of the whole system. The fill factor is a tool for displaying the main properties of this attractor in two dimensions and for determining diverse parameters for further analysis. A quantitative characterization of the system is possible by the distribution of correlation dimensions in the embedding space. The singular value decomposition (SVD) can be used to display and characterize individual degrees of freedom. These methods were applied to LDF time series from nine healthy controls and nine patients with Raynaud's phenomenon due to connective tissue disease. The fill factor and the SVD indicate qualitatively that in the controls vasomotion and heartbeat are the main influences on blood flow and act fairly independently of each other. In the patients there was a mixture of strong but irregular degrees of freedom. The mean and the maximal local correlation dimensions were significantly higher in the patient group. Nonlinear analysis of LDF time series provides additional information which cannot be detected using conventional approaches.

Adult↗
PubMed1999

Combined molecular and clinical approaches for the identification of families with familial adenomatous polyposis coli.

OBJECTIVE: Using an interdisciplinary clinical and molecular approach, the authors identified APC germline mutations in families with familial adenomatous polyposis (FAP). Correlation of mutation site with disease manifestation and the impact of molecular data on clinical proceedings were examined. SUMMARY BACKGROUND DATA: Germline mutations in the APC gene predispose to FAP. Established and proposed genotype-phenotype correlations as well as the influence of mutation site on surgical procedures have been reported. The predictive value of APC mutation analysis for disease manifestation and therapeutic decision making needs to be investigated further. METHODS: One hundred twenty-three kindreds of the local FAP registry were included in this study. CHRPE phenotype was defined as at least one large characteristic lesion or a total of four lesions in both eyes. APC mutations were identified by protein truncation test and automated DNA sequencing from patient lymphocyte DNA and RNA. RESULTS: APC germline mutations were identified in 85/123 families with FAP. They were located between codons 213 and 1581 of the APC gene and displayed distinct genotype-phenotype correlations. CHRPE status facilitated mutation analysis by discriminating regions of interest within the APC coding region. Severe manifestations of desmoids were restricted to mutations between codons 1444 through 1581. Whereas 91% (75/82) of at-risk persons were excluded as mutation carriers, APC germline mutations were detected before clinical examination in 9% (7/82) of at-risk persons. One patient agreed to endoscopy only after mutation detection. CONCLUSIONS: This study supports the feasibility of combined molecular and clinical screening of families with FAP and may provide a guideline for routine presymptomatic molecular diagnostics in a clinical laboratory.

Adenomatous Polyposis Coli↗
PubMed1999

The effect of compression therapy on venous haemodynamics in pregnant women.

OBJECTIVE: To study the influence of compression on the haemodynamics of the deep venous system in pregnancy. DESIGN: A prospective, observational study. SETTING: The phlebological unit of the department of dermatology at a university hospital. POPULATION: Fifteen pregnant women with no previous signs of chronic venous insufficiency. METHODS: Parameters of the venous pump function were assessed by strain-gauge plethysmography. Blood flow velocity, flow volume and vessel diameter in the superficial femoral vein were measured by duplex sonography. All examinations were performed with and without applied compression stockings (25-32 mmHg) at two different stages of gestation and after delivery. In addition, subjective symptoms were graded. RESULTS: Venous pump function improved and refilling time lengthened significantly when compression was used during gestation and postnatally. Duplex sonography showed an increase in blood flow velocity and flow volume in the superficial femoral vein with applied compression; the vessel diameter increased slightly. Subjective symptoms of the leg, graded on an arbitrary scale, were reduced by regular compression therapy. CONCLUSION: Our results indicate that compression improves the clinical symptoms of venous congestion and the venous haemodynamics of the legs during pregnancy and in the postpartum period. Thus, the regular use of compression during pregnancy and the puerperium may reduce the incidence of thromboembolic events.

Bandages↗
PubMed1999

[A simplified procedure for mechanical testing of lumbar spinal fixation implants].

On the basis of the current ASTM and ISO standard proposals, a simplified test procedure for spinal fixation implants has been developed. It comprises static and dynamic tests aimed at evaluating the stiffness and strength of various different internal implants. Different methods of mounting the pedicle screws to the test device are shown to significantly affect the characteristic values and failure mechanisms of the implants. The feasibility of the procedure was investigated by comparing 7 different internal fixation implants. The reproducible results revealed general differences associated with the material, dimensions and design, which latter in particular correlated with the specific failure mechanisms. For longer-term in situ duration, testing of these implants should be expanded to include an analysis of wear and corrosion properties.

Biomechanical Phenomena↗
PubMed1999

[Predictive molecular diagnosis and preventive surgical treatment of hereditary colorectal carcinoma: a new field of interest for surgical research].

Colorectal cancer is one of the most frequent cancers in the Western hemisphere. It seems to be well established that colorectal tumors develop as a result of an accumulation of inherited and/or acquired somatic mutational events in tumor suppressor genes and oncogenes. An increasing understanding of the molecular basis of the most prevalent colorectal cancer syndromes, such as hereditary nonpolyposis colorectal cancer (HNPCC) and familial adenomatous polyposis (FAP), is reflected by modifications in diagnosis and therapy. Therefore, strategies have been developed for predictive molecular diagnosis and preventive surgical treatment of colorectal cancer syndromes. In the future surgical research will participate in research and development in the field of molecular diagnosis of colorectal cancer and will then evaluate the clinical management concepts as a result.

Adenomatous Polyposis Coli↗
PubMed1999

Genetic alterations of the tumor suppressor gene PTEN/MMAC1 in human brain metastases.

The high mutation rate in advanced brain tumors, recent functional studies, and the high frequency of mutations in prostate metastases all strongly suggest that PTEN/MMAC1 alterations are involved in the formation of metastases. We searched for genetic alterations in the PTEN/MMAC1 gene in 56 consecutive brain metastases from various primary tumors by loss of heterozygosity (LOH), direct sequence analysis, and differential PCR analysis. The highest LOH rates were detected in metastases deriving from lung (67%) and breast (64%) cancers. Three (25%) of the eight detected inactivating mutations (one nonsense mutation, one splice-site mutation, one 11-bp deletion, and five homozygous deletions) were found in metastases originating from 12 different lung carcinomas, suggesting that PTEN/MMAC1 alterations may play a role in the progression of this tumor. With the exception of lung carcinomas, our findings indicate that genetic abnormalities of the PTENM/MMAC1 gene are only involved in a relatively small subset of brain metastases. However, the discrepancy between the high overall LOH rate (50%) and the low frequency of PTEN/MMAC1 mutation detection rate (14%) suggests the presence of one or more additional tumor suppressor genes on chromosome 10q.

Alleles↗