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121.
122.
Analysis of gene expression patterns in small amounts of human ventricular myocardium by a multiplex RNase protection assay 总被引:3,自引:0,他引:3
C. Mittmann Ursula Münstermann Joachim Weil Michael Böhm Stefan Herzig Christoph Nienaber Thomas Eschenhagen 《Journal of molecular medicine (Berlin, Germany)》1998,76(2):133-140
End-stage human heart failure is associated with changes in expression of steady-state messenger RNA (mRNA) levels. These
changes correspond to alterations in protein levels and myocardial function and may have clinical implications regarding etiology,
clinical state, or prognosis. However, analysis of mRNA levels in endomyocardial biopsies can be accomplished only by the
quantitative polymerase chain reaction, which is difficult to standardize. The aim of the study was to evaluate whether the
RNase protection assay is applicable to measure mRNAs of multiple genes simultaneously in small amounts of ventricular myocardium
comparable to myocardial biopsies. Total RNA was prepared from left ventricular myocardium from terminally failing hearts
with idiopathic (n=9) or ischemic cardiomyopathy (n=7) and from nonfailing control hearts (n=10). mRNA was measured by an optimized RNase protection assay for the β1-adrenoceptor, the stimulatory G protein α-subunit (Gsα), phospholamban, the calcium ATPase of the sarcoplasmic reticulum (SERCA), β-myosin heavy chain (β-MHC), and the atrial natriuretic
peptide (ANP). We extracted 10.7±2.1 μg total RNA from three myocardial biopsies taken in vitro. All of the six genes were
measurable in duplicate in a total of 7 μg RNA. mRNAs of β1-adrenoceptor, phospholamban, and SERCA were lower in failing than in nonfailing myocardium by 50%, 33%, and 42% respectively,
whereas β-MHC and Gsα mRNAs were unchanged. mRNA of ANP was expressed at high levels only in the failing myocardium, providing a highly specific
and sensitive marker for discriminating nonfailing and failing hearts. A direct comparison with ANP and Gsα levels obtained by Northern blot analysis with 7.5 μg total RNA showed a good correlation between the two methods. The RNase
protection assay is thus a suitable method for simultaneous measurements of multiple mRNA levels in human myocardial biopsies.
Changes in mRNA levels closely reflected those identified by other methods using larger amounts of RNA. Increased myocardial
ANP mRNA levels determined by the RNase protection assay may serve as a molecular marker of heart failure.
Received: 12 May 1997 / Accepted: 8 September 1997 相似文献
123.
Beneke R Beyer T Jachner C Erasmus J Hütler M 《European journal of applied physiology》2004,92(4-5):518-523
It is speculated that anaerobic metabolism is the predominant source of energy in karate kumite. However, no experimental proof is currently available. The metabolic cost and fractions of aerobic and anaerobic energy of karate kumite fighting were investigated. Ten male nationally or internationally ranked karateka [means (SD) age 26.9 (3.8) years, height 1.80 (0.08) m, mass 77.2 (12.8) kg] performed two to four fights scheduled and judged like a championship. Oxygen uptake was measured continuously with a portable spirometric device. Blood lactate was determined immediately before, and minute by minute after, each fight. Aerobic, anaerobic alactic and anaerobic lactic energy were calculated from oxygen uptake during the fight (VO2), the fast component of the post-fight oxygen uptake (VO2PCr) above resting values and changes in blood lactate concentration (Net-BLC), respectively. Altogether, 36 fights lasting 267 (61) s were analysed. The referees decisions caused an activity-to-break ratio of approximately 2:1. VO2, VO2PCr, and Net-BLC per fight were 165.3 (52.4) ml.kg–1, 32.2 (7.2) ml.kg–1and 4.2 (1.9) mmol.l–1; the overall energy cost above rest was 334.3 (86.3) kJ per fight. Fractions of aerobic, anaerobic alactic, and lactic energy sources were 77.8 (5.8)%, 16.0 (4.6)%, and 6.2 (2.4)%, respectively. The results indicate a high metabolic rate in karate kumite. However, the acyclic activity profile implies that aerobic metabolism is the predominant source of energy and there is anaerobic supplementation, mainly by high-energy phosphates. 相似文献
124.
125.
Epidemiological and Environmental Investigations of Legionella pneumophila Infection in Cattle and Case Report of Fatal Pneumonia in a Calf
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Massimo Fabbi Maddalena Castellani Pastoris Eugenio Scanziani Simone Magnino Luigi Di Matteo 《Journal of clinical microbiology》1998,36(7):1942-1947
A fatal pneumonia due to Legionella pneumophila was diagnosed in a young calf reared in a dairy herd located in northern Italy. Clinical symptoms consisted of watery diarrhea, hyperthermia, anorexia, and severe dyspnea. The pathological and histological findings were very similar to those observed in human legionellosis. Legionella pneumophila serogroup 1 (SG1) and SG10 were isolated from the calf’s lung, and L. pneumophila SG1 was isolated from the calf’s liver. L. pneumophila SG1 was also demonstrated in the lung tissue by immunofluorescence and immunohistochemical examinations. Nine of 10 L. pneumophila SG1 isolates belonged to the Olda subtype, and 1 belonged to the Camperdown subtype. A very low prevalence of antibodies to Legionella was detected in cows and calves reared in the same herd. Cultures of aqueous sediment of an old electric water heater which supplied hot water for the feeding of the calves yielded L. pneumophila SG1. Four of the colonies tested belonged to the Olda subtype. Ten clinical and four environmental isolates were examined for the presence of plasmids. Nine of them were also examined by pulsed-field gel electrophoresis assay, and the same patterns were found for L. pneumophila SG1 Olda strains isolated from the calf and from the electric heater. This is the first report of a documented case of a naturally occurring Legionella pneumonia in an animal. Cattle probably act as accidental hosts for legionellae, much the same as humans. 相似文献
126.
Michael Heming Xiaolin Li Saskia Räuber Anne K. Mausberg Anna-Lena Börsch Maike Hartlehnert Arpita Singhal I-Na Lu Michael Fleischer Fabian Szepanowski Oliver Witzke Thorsten Brenner Ulf Dittmer Nir Yosef Christoph Kleinschnitz Heinz Wiendl Mark Stettner Gerd Meyer zu Hörste 《Immunity》2021,54(1):164-175.e6
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127.
128.
H. Heuer Will Spijkers Thomas Kleinsorge Hanno van der Loo Christoph Steglich 《Experimental brain research. Experimentelle Hirnforschung. Expérimentation cérébrale》1998,118(3):381-392
We investigated the time course of the amplitude specification of rapid bimanual reversal movements (lateral displacements
on two digitizers). To this end we used the timed-response paradigm in which the response has to be initiated synchronously
with an auditory signal. Information about the required amplitudes was presented at various times before the synchronization
signal. Consistent with previous results, the progression of amplitude specification was reflected in the dependence of the
amplitudes of the reversal movements on the time interval between amplitude information and synchronization signal. Same or
different amplitudes for the hands were used to examine cross-talk at the programming level of the two-level model of intermanual
interference. The results indicate the existence of cross-talk in particular at short intervals between information about
amplitude and movement initiation. This is consistent with the notion that cross-talk between concurrent processes of amplitude
specification is transient and vanishes as the time available for motor programming increases.
Received: 28 August 1996 / Accepted: 10 July 1997 相似文献
129.
Schwarz C Hauser P Steininger R Regele H Heinze G Mayer G Oberbauer R 《Laboratory investigation; a journal of technical methods and pathology》2002,82(7):941-948
SUMMARY: In renal transplantation, postischemic acute renal failure (ARF) develops in more than 20% of patients. We investigated whether tubular epithelial cells obtained from donor kidneys without subsequent ARF express a different pattern of survival genes, compared with cells from kidneys exhibiting ARF. Donor kidney biopsy specimens were obtained before transplantation from eight recipients of cadaveric kidneys with primary graft function (CAD-PF), eight patients with biopsy-proven ARF without rejection (CAD-ARF), and eight recipients of living donor kidneys with primary graft function (LIV). One thousand proximal tubular epithelial cells per biopsy specimen were isolated by laser capture microdissection. Quantitative analysis of apoptosis and the apoptosis regulatory genes Bcl-2, Bcl-xL, and Bax were performed by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-digoxigenin nick-end labeling staining and real-time PCR, respectively. Primary cultures of human proximal tubular epithelial cells served as calibrator. The number of apoptotic cells was significantly higher in CAD-ARF compared with LIV and CAD-PF (1.5 +/- 1.1% [p < 0.05] vs. 0.3 +/- 0.2% vs. 0.4 +/- 0.2%; mean +/- SD). The apoptosis inhibitors Bcl-2 and Bcl-xL were significantly up-regulated in renal tubular cells of recipients without ARF compared with CAD-ARF. The ratios of Bcl-2/GAPDH normalized to calibrator were as follows: LIV 48 +/- 30, CAD-PF 38 +/- 55, and CAD-ARF 5 +/- 7 (p < 0.05). The corresponding ratios for Bcl-xL were as follows: LIV 6 +/- 6, CAD-PF 5 +/- 3, and CAD-ARF 1 +/- 1 (p < 0.05). No difference in the expression of the proapoptotic Bax could be observed. These data suggest that failure of proximal tubular cells to respond to injury by up-regulation of survival factors from the Bcl-2 family contributes to postischemic ARF in patients after cadaveric renal transplantation. 相似文献
130.