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  • 1
    ISSN: 1432-119X
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Radioactive in situ hybridization techniques or enzymatic detection procedures of hapten-modified human cytomegalovirus (HCMV) probes have been widely used for studying the infection of peripheral blood leukocytes with HCMV. This report describes significant improvements in terms of signal resolution which can be obtained by applying a highly sensitive fluorescence in situ hybridization (FISH) technique in conjunction with a large subgenomic HCMV DNA probe. Three cosmid clones spanning 119.1 kb of the HCMV genome (230 kb) were used to construct the digoxigenin-11-dUTP-labeled probe which was found to be superior to a total HCMV probe representing the entire genome. Crucial hybridization parameters were analyzed systematically in order to ensure optimal resolution power and sensitivity. The protocol was successfully applied to HCMV-infected fibroblasts and peripheral blood leukocytes of 12 transplant patients and unambiguously facilitated the precise intracellular localization of HCMV genomes in infected cells. Because of its excellent resolution properties, accompanied by the virtual absence by any background staining, we recommend the use of this protocol as a sensitive approach for further virological analyses of the interactions between HCMV and peripheral blood leukocytes at the single-cell level.
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  • 2
    ISSN: 1432-1076
    Keywords: Intravascular Coagulation ; Neonatal Thrombosis ; Perinatal Consumption Coagulopathia ; Shock (Inadequate Capillary Perfusion) ; Perinatal Death
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Zusammenfassung Bei drei Neugeborenen fand sich eine Thrombose der peripheren Extremitätengefäße als morphologisches Teilsymptom einer generalisierten plasmatischen Hyperkoagulabilität und disseminierten intravasalen Gerinnung der Neonati unter der Geburt und in der Perinatalperiode. Fibrinreiche intravasale Mikrothromben in den Extremitätengefäßen können morphologisches Äquivalent einer intrauterin eingeleiteten plasmatischen Hyperkoagulabilität und bereits intrauterin nachfolgenden disseminierten intravasalen Gerinnung darstellen. Eine disseminierte intravasale Gerinnung post natum kann daneben im Verlauf einer intrauterin begonnenen, perinatal fortbestehenden generalisierten plasmatischen Hyperkoagulabilität in der unmittelbaren Perinatalperiode auftreten und durch den hypoxämieprovozierenden Einfluß eines Membranensyndroms perpetuiert und intensiviert werden. Hypoxämie und Acidose können nach rezidivierendem Atemstillstand auch unabhängig von asphyxieprovozierenden Geburtskomplikationen eine generalisierte plasmatische Hyperkoagulabilität und nachfolgende disseminierte intravasale Gerinnung induzieren. An Hand der drei vorliegenden Fälle werden die pathogenetischen Beziehungen zwischen den Faktoren Hypoxämie und Acidose in der Blutbahn der Nascituri und Neonati, plasmatische Hyperkoagulabilität unter Thrombocytopenie und Verbauch von Gerinnungsfaktoren in utero, hämorrhagische Diathese unter der Geburt, extravasale Polymerisation der intravasal entstehenden Fibrinmonomere zu perinatalen pulmonalen hyalinen Membranen einerseits und der intravasalen Polymerisation der Monomere zu perinatalen disseminierten intravasalen Gerinnseln mit Thrombose peripherer Extremitätengefäße andererseits diskutiert.
    Notes: Abstract Three out of four newborn infants (premature twins, one case of prolonged delivery and one case of relapsing respiratory arrest) showed microthrombi in the vessels of the extremities as morphological symptoms of a generalized plasmatic hypercoagulability, consumption of clotting factors, and disseminated intravascular coagulation which takes place in the newborns during birth and during the perinatal period. The thrombi in the extremities are regarded as clinical manifestation of a generalized microthrombosis which involves the vascular system of many organs, i.e. liver, lungs, spleen, intestine, suprarenal glands, and kidneys. Fibrin-rich intravascular microthrombi in the vessels of the extremities are considered as morphological equivalents of a generalized hypercoagulability and subsequent disseminated intravascular coagulation. They are already initiated in utero following intra-uterine asphyxia with hypoxemia and acidosis in the capillary microcirculation (inadequate capillary perfusion). Disseminated intravascular coagulation can also occur in the immediate perinatal period due to a generalized hypercoagulability of the plasma, which had been initiated in utero and persisted during the perinatal time without having been already converted in utero into a disseminated intravascular microthrombosis. Hypercoagulability can be perpetuated and intensified by a membrane syndrome in newborn infants with respiratory distress syndrome. The pathogenetic relations between hypoxemia and acidosis in the microcirculation of nascituri and newborns, hypercoagulability of plasma with thrombocytopenia and with consumption of clotting factors in utero, hemorrhagic diathesis during birth, disseminated intravascular coagulation in stillbirths and in newborn infants, and the respiratory distress syndrome with hyaline membranes of the lung are discussed.
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  • 3
    ISSN: 1432-8798
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Summary Although viremia is an integral part of the pathogenesis of human cytomegalovirus (HCMV) disease, the interaction between HCMV and circulating leukocytes of actively infected patients remains an area of uncertainty. It is still a matter of dispute, whether leukocytes support viral replication with subsequent production of infectious virus. In a new approach we developed and applied a sensitive fluorescence in situ hybridization assay for the precise intracellular localization of HCMV genomes in leukocytes. It was shown that in vivo HCMV genomes were exclusively localized in the cytoplasm of leukocytes, indicating that the majority of these cells are virus carriers or abortively infected. Though this method easily detects single copy genes in metaphase chromosomes, the number of HCMV DNA positive leukocytes was significantly lower than the number of HCMV pp65 antigen positive cells. In relation to the pp65 antigen positive cells, only 1–4% of these cells were DNA positive. In addition, the much lower frequency of HCMV immediate early antigen positive leukocytes in comparison to the pp65 antigen positive cells and the impossibility of detecting other viral antigens support the hypothesis that the origin of pp65 found in leukocytes results mainly from protein uptake.
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  • 4
    ISSN: 1433-0474
    Keywords: Schlüsselwörter Anomaler Abgang der rechten Koronararterie ; Bland-White-Garland-Syndrom ; Angeborene Herzfehler ; Key words Anomalous origin of the right coronary artery ; Bland-White-Garland-Syndrome ; Congenital heart disease
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary Anomalous origin of the right coronary artery from the pulmonary trunk is rare, however, it is of considerable clinical significance. Discussion: We report of the findings in an asymptomatic child with anomalous origin of the right coronary artery from the pulmonary trunk and shortly review this entity.
    Notes: Zusammenfassung Der fehlerhafte Abgang der rechten Koronararterie aus der Pulmonalarterie ist selten, jedoch von erheblicher klinischer Relevanz. Diskussion: Wir berichten über die Befunde eines asymptomatischen Schulkinds mit diesem Krankheitsbild und geben einen Überblick über diese Fehlbildung.
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  • 5
    ISSN: 1591-9528
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Zusammenfassung Im Gegensatz zur Rückresorption von Glucose, Orthophosphat und Sulfat bleibt die Ascorbinsäure-Rückresorption der Katze bei geringer Diurese mit steigender Belastung im Bereich annähernd konstanter Werte von rund 6 mg/100 ml Filtrat stehen. Osmotische Diurese führt zu einer Verminderung der Ascorbinsäure-Rückresorption. Da sich mit zunehmender Plasmakonzentration auch bei stärkster osmotischer Diurese annähernd konstante — wenn auch erniedrigte — Rückresorptionswerte einstellen, könnte man bei der Katze von „diureseabhängigen tubulären Maxima“ für Ascorbinsäure sprechen. Im Gegensatz zu Ascorbinsäure wies die Rückresorption von Glykokoll und racemischem Alanin bei der Katze keine Beschränkung im Sinne tubulärer Maxima auf. Osmotische Diurese führte auch bei diesen Aminosäuren und besonders deutlich bei d-Alanin zu einer Verminderung der Rückresorption. Zur Erklärung der Diureseabhängigkeit wird — neben der Annahme teilweiser oder ausschließlicher Rückdiffusion — auf die morphologischen Nierenveränderungen bei osmotischer Diurese hingewiesen.
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  • 6
    ISSN: 1398-9995
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background:  A polymorphism in the promoter region of the CD14 gene, C-159T, has been shown to be associated with increased levels of soluble CD14 (sCD14) and decreased serum immunoglobulin E (IgE) and the expression of a more severe atopic phenotype in previous studies.Methods:  To test if these associations are consistently found in different populations and different age groups, we genotyped 2048 children of different age groups as well as 888 adults from different regions of Germany for the CD14 C-159T polymorphism.Results:  While an association between this promoter polymorphism and levels of sCD14 could be confirmed in our study population (CC: 1017 ng/ml vs TT: 1370 ng/ml, P = 0.03), no association between CD14 C-159T genotypes and IgE levels or the prevalence of atopic diseases was seen.Conclusions:  The lack of association between CD14 genotypes and IgE as well as atopic outcomes in this large German study population seems to indicate that CD14 genotypes may not directly be involved in the development of allergies during childhood.
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  • 7
    ISSN: 1365-3083
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Reliable quantification of cytokine mRNA expression is an important technique for analyzing immune responses. Up until now, little to no information has been available as to whether different mRNA quantification techniques lead to similar results. Recently, real time quantitative reverse transcriptase (RT)-PCR using SYBR® Green I as a double stranded DNA specific dye has been introduced. This novel method enables simple and rapid measurement of PCR product accumulation during the log-linear reaction phase and obviates the need for expensive hybridization probes. Here, we analyzed murine gamma interferon (IFN)-γ mRNA expression in splenocytes by this technique in comparison to semiquantitative noncompetitive RT–PCR, Northern blot analysis, and ELISA after stimulation of the cells with interleukin (IL)-12, IL-18 and a combination of both cytokines. The results clearly show that all of the techniques detect differences in the IFN-γ gene expression induced by these distinctive stimuli qualitatively exactly in the same order. However, real-time kinetic RT–PCR offers several advantages, notably its high sensitivity that allows the detection of basal IFN-γ mRNA expression in unstimulated samples. In addition it provides the lowest interassay variability of all techniques investigated. Finally, the gene expression measured by this method eliminates any post-PCR manipulations because the PCR product identification can be easily performed by melting curve analysis.
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  • 8
    ISSN: 1365-2222
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Medicine
    Notes: Background Toll-like receptor 9 (TLR9) is a pattern-recognition receptor that detects unmethylated CpG motifs prevalent in bacterial and viral DNA. TLR9 stimulation is a key event after bacterial infection, triggering innate immunity and T-helper type 1 skewed adaptive immunity. Synthetic CpG-oligodeoxynucleotides (CpG-ODNs) represent a promising and novel class of immune adjuvants for allergy treatment, vaccination, and cancer therapy. However, common functional TLR9 gene variants could interfere with the clinical utilization of CpG-ODN in immunotherapy. Recently, a possible association of TLR9 polymorphism C-1237T with asthma has been reported.Objective The aim of the present study was to investigate whether TLR9 polymorphisms or haplotypes have functional relevance and are associated with atopy.Methods We genotyped five common TLR9 single-nucleotide polymorphisms (SNPs) in promoter, exon, and intron regions of the gene in 527 healthy blood donors, and estimated four common haplotypes. The total IgE and specific IgE levels against the most common aeroallergens were measured (n=303). IFN-α production by plasmacytoid dendritic cells (pDCs) was analysed after stimulation with TLR9 ligand CpG-ODN (n=220).Results No significant influence of common TLR9 polymorphisms and haplotypes on the total and specific IgE levels was found. Functional analysis of CpG-ODN-induced IFN-α did not indicate a significant role for common TLR9 gene polymorphisms in TLR9 function.Conclusion We conclude that common genetic differences in the TLR9 gene exert no major influence on allergy susceptibility, and are unlikely to have on impact on clinical application of CpG-ODNs.
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  • 9
    ISSN: 1432-1971
    Keywords: Key words: Duchenne — Becker — X-chromosomal muscular dystrophy — Dilated cardiomyopathy
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Notes: Abstract. In order to screen for cardiac abnormalities, we prospectively studied 15 patients (age 8–25 years, mean 15.5 years) with Duchenne's (DMD) (n= 9) and Becker's (BMD) (n= 6) muscular dystrophy using the echocardiogram. Data were compared to a control group of 92 healthy individuals (age 7.9–25 years, mean 14.3 years). Left ventricular filling in diastole showed a different pattern when comparing echocardiographic Doppler results in patients and controls: Patients had lower peak velocity of early left ventricular diastolic filling (E-vmax)(P 〈 0.0001) and smaller time velocity integral of the E-wave (E-tvi)(P 〈 0.0001). In contrast, the atrial component (A-vmax, A-tvi) of diastolic filling in DMD/BMD showed no significant difference to controls. The mean area of the mitral valve orifice was significantly larger in patients (P 〈 0.0001) without presence of mitral regurgitation. Systolic left ventricular function was significantly impaired in the DMD/BMD group; we found lower heart rate corrected fiber shortening velocity VCFc (P 〈 0.001) and higher peak systolic wall stress (P 〈 0.001) in DMD/BMD. In 8 of 15 patients, peak systolic wall stress was above 95th percentile of controls. In 6 of 15 patients, VCFc was lower than the 5th percentile of controls. Systolic and diastolic myocardial impairment was found even in young patients and at low stages of disability—equally among patients with DMD or BMD. Diastolic left ventricular impairment predominantly affected the early diastolic filling, but atrial compensation was poor. Peak systolic wall stress measurements were particularly useful in patients with CMP, reflecting the left ventricular afterload.
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  • 10
    Electronic Resource
    Electronic Resource
    Springer
    Lung 131 (1965), S. 331-337 
    ISSN: 1432-1750
    Source: Springer Online Journal Archives 1860-2000
    Topics: Medicine
    Description / Table of Contents: Summary We followed the destiny of 131 patients after their lung extirpation in our hospital between 1953 and 1964. The lung extirpation was performed upon them because of the specific highest damage of one lung. A thorough analysis of any possible premature or subsequent complication of our own patients was made. Particularly among one part of the patients the ventilation could often be examined even after years. In a questioning about their further destiny all the alive patients could be comprehended. We are glad to state that now more than one half of the patients were able to work, whereas at one time, they used to die or only after the chemotherapy to be invalids for years in isolation hospitals.
    Notes: Zusammenfassung Wir gingen dem Schicksal von 131 Tuberkulosekranken mit einer destroyed lung nach, die zwischen 1953 und 1964 in unserem Hause pneumonektomiert wurden. Die möglichen Früh- und Spätkomplikationen wurden eingehend an Hand der eigenen Zahlen analysiert. Besonderes Gewicht wurde auf die Bronchusstumpfinsuffizienz und das postoperative Empyem gelegt. Bei einem Teil der Fälle konnte die Ventilation oft noch nach Jahren überprüft werden. In einer katamnestischen Befragung konnte das Schicksal aller Pat. erfragt werden. Es ergab sich die erfreuliche Tatsache, daß die Hälfte aller am Leben befindlichen Patienten heute noch arbeitsfähig sind. Nur bei 12 Pat. besteht ein Tb-positives Sputum. Besonderes Gewicht bei der Langzeitnachbehandlung der Pneumonektomierten sollte neben der regelmäßigen Röntgenkontrolle auf den Zustand der Bronchien gelegt werden. Die Entstehung eines obstruktiven Emphysems sollte soweit als irgend möglich verhindert werden.
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