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Biomedical subjects

M Prinz

Publications and source records attributed to M Prinz.

At least 55 records · Page 3Linked to original sources

Mouse brain microglia express interleukin-15 and its multimeric receptor complex functionally coupled to Janus kinase activity.

The cytokine, interleukin (IL)-15, and the T cell growth factor, IL-2, exhibit a similar spectrum of immune effects and share the IL-2 receptor (IL-2R) subunits IL-2Rbeta and IL-2Rgamma for signaling in hematopoietic cells. Numerous neuroregulatory activities of IL-2 have been suggested, but its expression in the normal central nervous system (CNS) is apparently very low and regionally restricted. We show by RNA and protein detection that IL-15, its specific receptor molecule, IL-15Ralpha, and the signal-transducing receptor subunits, IL-2Rbeta and IL-2Rgamma, are constitutively present in various regions of the developing and adult mouse brain. We further demonstrate, also at the single-cell level, that IL-15 and the components for IL-15Ralpha/IL-2Rbetagamma receptors are expressed by microglia. Tyrosine phosphorylation data are presented showing that IL-15 signaling in microglia involves Janus kinase 1 activity. At doses of 0.1-10 ng/ml, IL-15 affected functional properties of these cells, such as the production of nitric oxide, and supported their growth in culture, suggestive of a role as an autocrine growth factor. Microglial IL-15 could thus play a pivotal role in the CNS and may participate in certain CNS and neuroendocrine functions previously ascribed to IL-2.

Animals↗

Endothelin-induced calcium signaling in cultured mouse microglial cells is mediated through ETB receptors.

Microglial cells are the intrinsic immunocompetent cells of the central nervous system, which are activated by brain tissue damage. In this paper we investigated the ability of endothelins (ETs), which are potent vasoconstrictors, to induce intracellular calcium signals in cultured microglia cells. Both endothelin-1 and endothelin-3 increased intracellular Ca2+ concentration ([Ca2+]i). These [Ca2+]i transients were mimicked by BQ3020, an ETB receptor agonist and blocked by BQ788, a selective ETB antagonist, respectively. The calcium signals induced by the endothelins persisted in Ca(2+)-free media. Transcripts encoding the ETB receptor were detected in purified microglial cultures and cDNA fragments derived from ETB receptor mRNA were amplified from 9% of electrophysiologically characterized microglial cells by the use of single-cell RT-PCR.

Animals↗

Multiplexing of Y chromosome specific STRs and performance for mixed samples.

A combination of four Y-specific polymorphic STR loci was amplified simultaneously using fluorescently labeled primers. Multiplex conditions required optimization to eliminate constant bands and amplification products for female DNA. A series of experiments was carried out for mixtures of DNA from two males, and from male and female individuals for the Y-specific STRs and an autosomal locus. For the male/male mixtures amplified with the Y specific system, and amplified for an autosomal locus, the minor component in the mixture could only be identified up to a ratio of 1:10, 1:50 respectively. In male/female DNA mixtures the Y STR alleles could be identified for the highest ratio tested, 400 pg male in DNA in 800 ng female DNA which amounts to a ratio of 1:2000.

Alleles↗

Evaluation of Y-chromosomal STRs: a multicenter study.

A multicenter study has been carried out to characterize 13 polymorphic short tandem repeat (STR) systems located on the male specific part of the human Y chromosome (DYS19, DYS288, DYS385, DYS388, DYS389I/II, DYS390, DYS391, DYS392, DYS393, YCAI, YCAII, YCAIII, DXYS156Y). Amplification parameters and electrophoresis protocols including multiplex approaches were compiled. The typing of non-recombining Y loci with uniparental inheritance requires special attention to population substructuring due to prevalent male lineages. To assess the extent of these subheterogeneities up to 3825 unrelated males were typed in up to 48 population samples for the respective loci. A consistent repeat based nomenclature for most of the loci has been introduced. Moreover we have estimated the average mutation rate for DYS19 in 626 confirmed fatherson pairs as 3.2 x 10(-3) (95% confidence interval limits of 0.00041-0.00677), a value which can also be expected for other Y-STR loci with similar repeat structure. Recommendations are given for the forensic application of a basic set of 7 STRs (DYS19, DYS3891, DYS389II, DYS390, DYS391, DYS392, DYS393) for standard Y-haplotyping in forensic and paternity casework. We recommend further the inclusion of the highly polymorphic bilocal Y-STRs DYS385, YCAII, YCAIII for a nearly complete individualisation of almost any given unrelated male individual. Together, these results suggest that Y-STR loci are useful markers to identify males and male lineages in forensic practice.

DNA Mutational Analysis↗

Chromosome Y microsatellites: population genetic and evolutionary aspects.

By means of a multicenter study, a large number of males have been characterized for Y-chromosome specific short tandem repeats (STRs) or microsatellites. A complete summary of the allele frequency distributions for these Y-STRs is presented in the Appendix. This manuscript describes in more detail some of the population genetic and evolutionary aspects for a restricted set of seven chromosome Y STRs in a selected number of population samples. For all the chromosome Y STRs markedly different region-specific allele frequency distributions were observed, also when closely related populations were compared. Haplotype analyses using AMOVA showed that when four different European male groups (Germans, Dutch, Swiss, Italians) were compared, less than 10% of the total genetic variability was due to differences between these populations. Nevertheless, these pairwise comparisons revealed significant differences between most population pairs. Assuming a step-wise mutation model and a mutation frequency of 0.21%, it was estimated that chromosome Y STR-based evolutionary lines of descent can be reliably inferred over a time-span of only 1950 generations (or about 49,000 years). This reduces the reliability of the inference of population affinities to a historical, rather than evolutionary time scale. This is best illustrated by the construction of a human evolutionary tree based on chromosome Y STRs in which most of the branches connect in a markedly different way compared with trees based on classical protein polymorphisms and/or mtDNA sequence variation. Thus, the chromosome Y STRs seem to be very useful in comparing closely related populations which cannot probably be separated by e.g. autosomal STRs. However, in order to be used in an evolutionary context they need to be combined with more stable Y-polymorphisms e.g. base-substitutions.

Biological Evolution↗

The growth of non-pyramidal neurons in the primary motor cortex of man: a Golgi study.

In Golgi-Cox-impregnated motor cortex (regio precentralis) of six children (36 weeks of gestation to 13 month after birth) four types of interneurons of laminae II-VI have been investigated qualitatively and quantitatively and finally, compared with interneurons of a child (3 month of age) with Down's Syndrome. All cell measurements have been made by using a computerized digital morphometric system. In order to study the developmental stage of interneurons in our cases we statistically compared analogue neuron types in the brains. As parameters for characterizing non-pyramidal neurons served the pattern of dendritic branching, dendritic length for all dendritic orders within single dendritic fields, and of the whole neuron. The quantitative morphological study of dendritic arborizations revealed that bitufted and large multipolar interneurons had significantly more branching material in the postnatal brains than their prenatal counterparts. These differences were due to the increase of both length and number of branches during perinatal period. Such constant increment in dendritic span and branching provides larger receptive areas and that should improve the development of connections in functional intracortical columns. The quantitative parameter area of dendritic fields and number of branching points of the brain with Down's syndrome revealed in comparison with normal brain of the same age differences such as lower dendritic areas and a higher amount of branching points.

Dendrites↗

Resolution of apolipoprotein B repeat unit position variants on agarose, denaturing, and native polyacrylamide gels.

Utilizing three different electrophoresis systems for the separation of variable number of tandem repeats (VNTR) alleles of the high variable region 3' from the apolipoprotein B (Apo B) locus, it could be shown that apparent allele subtypes on native polyacrylamide gels comigrated with the regular ladder alleles on agarose and denaturing polyacrylamide gels. Two subtypes, one with a shift towards the cathode and one shifted towards the anode, and the corresponding ladder alleles were sequenced in order to determine the molecular basis for this aberrant electrophoretical mobility. Subtype and ladder alleles displayed the same number of repeat units but differed in the number and order of different types of repeats. Depending on the variation in the region of pure AT repeat units, the potential secondary structures of hairpin loop formation differed for subtypes and ladder alleles, which is thought to explain the shift in electrophoretic mobility on native polyacrylamide gel electrophoresis.

Apolipoproteins B↗

Consonant clusters in child phonology and the directionality of syllable structure assignment.

The production of target consonant clusters at early stages of acquisition is analysed from a phonological representational perspective. The data stem from five normal monolingual German and four normal monolingual Spanish children at ages from 0;9 to 2;1, observed in naturalistic settings. At the beginning stages, target clusters are reduced to a single consonantal position, due to lack of branching of the syllabic constituents. This finding coincides with other results in the literature, which have in general been explained by means of universal principles. Nevertheless, there is an essential difference between the German and the Spanish data: German children tend to prefer the first consonant and Spanish children the second one. This difference can only be explained in terms of parameterization of syllabification, which in German takes place from left to right and in Spanish from right to left. At later stages, when clusters begin to be produced with two consonantal positions, they offer evidence for the beginning of branching of syllabic constituents, due to parameterization, and for the chronological order of the setting of the subsyllabic parameters. Our data offer evidence in favour of the following acquisitional hierarchy: CV > CVC > CVCC > CCVCC.

Child↗

Clear cell meningioma: report of a spinal case.

A sacrococcygeal case of clear cell meningioma in a 38-year-old man is reported. Compared to the seven spinal clear cell meningiomas described so far, this case demonstrated the highest recurrency rate. Multiple recurrences were most likely due to the extensive infiltrative growth pattern of the tumor hindering complete microscopic surgical resection. Histologic parameters were not relevant to predict recurrences, since cellular anaplasia was lacking and growth fraction was low in the first operation and all subsequent recurrences. In view of what is now known of clear cell meningiomas, the tumor showed all histologic and immunohistochemical features which are typical for this tumor entity. However, the tumor was first diagnosed by capable pathologists and neuropathologists as fibroma-like mesenchymal tumor, metastatic renal carcinoma, chordoma, chondroma untypical osteosarcoma, and microcystic meningioma. Correct diagnosis was made by electron microscopy which has revealed comparable findings to those of other rare electron microscopical cases reported so far; high content of cytoplasmic glycogen diffusely distributed throughout the cytoplasm or aggregated within vacuoles, intermediate filaments, desmosomes, interdigitation of cell membranes and large amounts of collagen fibers, some of which were of the giant amianthoid type. Since meningiomatous features of clear cell meningiomas are not obvious light microscopically and the tumors may be confused with nonmeningiomatous neoplasms, electron microscopical investigation is highly recommended in each case of suspected clear cell meningioma.

Adult↗

High sensitive DNA typing approaches for the analysis of forensic evidence: comparison of nested variable number of tandem repeats (VNTR) amplification and a short tandem repeats (STR) polymorphism.

The approach of using nested primers for the APO B variable number of tandem repeats (VNTR) increases the sensitivity of the polymerase chain reaction (PCR) to single cell level. Different experiments and a comparison to the short tandem repeats (STR) system VWA were carried out, to determine the applicability of this method to forensic samples. Nested amplification of the Apo B VNTR was affected by a strong tendency towards preferential amplification of the shorter alleles. This phenomenon was observed for DNA quantities as low as 100 pg and impaired, depending on the allele length, the results for mixed samples. As expected, VWA polymorphism showed less preferential amplification. The high sensitivity of both PCR systems is accompanied by an increased susceptibility to contamination. Using artificially contaminated bloodstains, the bloodstain genotype, the contamination or both genotypes could be found on one piece of evidence. Here a single analysis can lead to an incorrect result. Therefore a strategy for obtaining reliable results should consist of multiple stain extractions and the amplification of different stepped dilutions of the DNA solution.

Base Sequence↗

DNA typing of urine samples following several years of storage.

The possibility of typing DNA polymorphisms on urine samples was investigated in a controlled storage experiment and for samples that were 1-7 years old. Female urine samples showed a higher amount of epithelial cells and therefore a higher DNA yield. Employing the polymerase chain reaction, specific amplification results were achieved for all samples over a 6 month storage period. The microscopical examination of the samples revealed not only differing degrees of contamination with bacteria, yeasts and fungi, but also the presence of still intact epithelial cells. Only 20% of the male samples and 32% of the female samples yielded specific amplification results. By separating the human cells from the contaminating organisms prior to DNA extraction, the number of successfully typed samples could be improved to 35% of the male and 77% of the female samples. This result confirms that excess amounts of co-extracted non-human DNA can inhibit the specific amplification of human target sequences.

Cell Count↗

Analysis of creatine kinase, CK-MB, myoglobin, and troponin T time-activity curves for early assessment of coronary artery reperfusion after intravenous thrombolysis.

BACKGROUND: Thrombolysis has become the standard therapeutic approach in patients with acute myocardial infarction. To identify patients who may benefit from early invasive procedures, reliable noninvasive assessment of success or failure of thrombolytic therapy is mandatory. METHODS AND RESULTS: In a prospective study in 63 consecutive patients undergoing thrombolysis for their first myocardial infarction, serial measurements of creatine kinase (CK), its isoenzyme CK-MB, myoglobin, and troponin T were done to determine their value for noninvasive prediction of coronary artery patency. Blood samples were drawn every 15 minutes during the first 90 minutes, every 30 minutes during the first 4 hours, every 4 hours during the first 24 hours, and every 8 hours during the first 72 hours. The perfusion status of the infarct-related artery was assessed angiographically 90 minutes after initiation of thrombolysis. For each marker, time to its peak concentration and its early initial slope (start of thrombolysis to 90 minutes thereafter) were determined. Areas under receiver operator characteristic (ROC) curves were 0.83, 0.76, 0.82, and 0.80 for maxima of CK, CK-MB, myoglobin, and troponin T, respectively (p = NS by univariate Z test). The corresponding values for early slopes of CK, CK-MB, myoglobin, and troponin T were 0.79, 0.82, 0.89, and 0.80 (p = 0.23 for comparison between myoglobin and CK-MB; p = 0.07 between myoglobin and CK). Sensitivity, specificity, and positive and negative predictive values regarding noninvasive prediction of coronary artery patency after 90 minutes were 80%, 82%, 95%, and 61% for time to CK maximum; 91%, 77%, 91%, and 77% for time to myoglobin maximum; 87%, 71%, 89%, and 67% for early CK slope; and 94%, 88%, 94%, and 82% for myoglobin slope, respectively. When myoglobin slope was assessed together with other clinical reperfusion markers (resolution of chest pain or ST segment elevation, occurrence of reperfusion arrhythmias) by logistic regression analysis, only the myoglobin slope was an independent predictor of coronary artery patency (p < 0.0001). CONCLUSIONS: With regard to noninvasive prediction of coronary artery patency after thrombolytic therapy, measurement of the early initial slopes of the serum markers within only 90 minutes after the initiation of therapy is as accurate as the determination of the time to their peak concentration. Compared with the other markers examined, myoglobin appears to have advantages because of its earlier rise, yielding a better negative predictive value and a higher area under the ROC curve for determination of its early initial slopes.

Adult↗

[Experimental studies on intraligamental anesthesia].

In an experimental study on animals the correlation between the pressure created by periodontal ligament injection and the distribution pattern of the anesthetic solution was examined. It was possible to demonstrate the penetration following intraligamental injection by scintiscanning of the mandibular bone. A correlation between pressure and distribution could not be established.

Anesthesia, Dental↗

The effect of various stain carriers on the quality and quantity of DNA extracted from dried bloodstains.

Bloodstains were made with 200 microliters blood on each of 11 different common substrates to examine the effect of the stain carrier on the amount and quality of DNA recoverable. High-molecular-weight DNA was extracted from all samples after 2 days. The yield of DNA from each sample varied considerably, not only between the different stain carriers but also within a given category. With a DNA yield of up to 10 micrograms, paper, glass, nylon, wood, smooth leather and wool gave the best results, followed by blue denim and wallpaper (up to 6 micrograms), cotton fabric and carpeting (up to 4 micrograms) and suede (up to 2 micrograms). For several stain carriers the DNA-containing solution was contaminated by chemical substances, which in the case of the blue denim, suede, and carpet samples inhibited the digestion of the DNA with restriction enzymes and prevented DNA typing. The different textures of the stain carriers tested and (as for varying yields on the same carrier) the differing degree of loss of DNA during extraction and the physiological variation in the number of leukocytes in human blood are discussed as possible reasons for the wide range of variation in the amounts of DNA it was possible to extract.

Blood Stains↗

[Possibilities for individual identification of hair roots using DNA probes].

High-molecular weight DNA was extracted from hair roots. The obtained DNA was digested with Hae III and Msp I and transferred to nylon membranes by Southern blotting. The possibility of applying a probe for sex determination (pHY2.1) a "single-locus"-probe (YNH24) as well as a "multi-locus"-probe (M13) was examinated as a function of the amount of hair being supplied. In the laboratory experiment a single hair root was enough for a sex determination or individual identification. Because of methodical difficulties in practice this kind of profiling should only be done with several hair roots.

Adolescent↗

DNA extraction from bloodstains in respect to age and stained substrate.

The amount and the quality of DNA that could be extracted from bloodstains equivalent to 200 microliters blood were examined after 3, 6, 9, 12, 15, 18 and 21 days of storage under dry or humid conditions at room temperature. DNA was also extracted from two days old 200 microliters bloodstains on different stain carriers. The amount of DNA that could be extracted showed a dependency on all the parameters that were examined. On carriers with a rough surface structure, where the blood can soak in, the resolving of leukocytes was impaired and the DNA recovery rate was low, e.g. carpeting and suede. The DNA yield was higher for substrates with a smooth surface, e.g. paper, glass and smooth leather. The aging experiments revealed that for the stains stored under dry conditions, there was no decrease in the DNA yield for stains on glass, while the amount of DNA that could be extracted became less for older stains on cotton. Under humid conditions the DNA yield was high and did not decrease for both glass and cotton fabric. The quality of the DNA was not impaired by aging or storage conditions, but for several stain carriers the DNA was contaminated by chemical substances which inhibited restriction enzyme digestion and successful DNA-typing.

Aging↗

[Methodologic variants of DNA blood stain analysis: extraction in agarose and digoxigenin marking].

DNA was extracted from 1 microliter bloodstains on glass, cotton cloth and paper using proteinase K digestion in a solution containing low gelling temperature agarose. This method is advantageous for small amounts of source material because the purifying procedures of other methods that contain a risk of loss of DNA, are avoided. The recovered DNA was digested with Hae III and after Southern blotting, a Digoxigenin labeled probe (Amprobe, human Y-chromosome specific repeat) was successfully used for sex determination. With this non-radioactive method quantities of DNA smaller than 1.5 pg were detected.

Blood Grouping and Crossmatching↗