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Biomedical subjects

B Herrmann

Publications and source records attributed to B Herrmann.

At least 37 records · Page 2Linked to original sources
PubMed2000

Trace analysis of endogenous and exogenous biomolecules from archaeological skeleton materials.

An analytical procedure was developed for the determination of pharmacologically active substances in archaeological skeleton materials. In comparative model studies, added ("spiked") test biomolecules of varying chemical behaviours were extracted from sample matrices and percentage of the analytes recovered were estimated using gas chromatography/mass spectrometry (GC/MS) or high performance liquid chromatography (HPLC). For the sterols and steroids studied, several organic solvents were appropriate. Extraction yields for the alkaloid nicotine, representing non-endogenous basic agents, were increased by alkalizing with triethylamine or by extracting with a two-phase system consisting of an alkaline aqueous and an organic layer (toluene). The flavonol quercetin was extractable only in an acidic environment. In a screening for native biomolecules using GC/MS, nicotine was identified in individual samples, in addition to lipophilic substances such as the endogenous cholesterol and its degradation products and two phytosterols which may have migrated into the bones from the surrounding soil.

Alkaloids↗
PubMed2000

Differentiation between occupational and dietary-related intake of barium in the skeleton.

On the basis of the reconstruction of general nutritional patterns by trace element data on historic populations, an examination on occupational Ba intakes in the skeleton was carried out for a historical metallurgy population. The concentrations of the trace elements Ba, Sr, and Pb as well as the matrix element Ca were determined by AAS analysis. Their correlations within the population were analysed statistically. Evidence of a correlation between Ba and Sr concentrations and the lack of a correlation between Ba and Pb concentrations is interpreted as evidence for a probably solely dietary-related Ba intake in the skeleton. Ba intake from working in ore smelting plants is therefore unlikely or at least insignificant. These conclusions are supported by analysis results of modern food items from the catchment area of the historical population. A sufficient Ba supply through rye was determined, thereby explaining the concentration of Ba in the bone material of the historic population.

Barium↗
PubMed1999

Optimized DNA extraction to improve reproducibility of short tandem repeat genotyping with highly degraded DNA as target.

The reproducibility of short tandem repeat (STR) genotyping of highly degraded DNA is often reduced due to artifacts generated during polymerase chain reaction (PCR) amplification. The frequency and amount of these artifacts are related to the quality and quantity of the DNA amplified. Consequently, the aim of this investigation was the optimization of DNA extraction to increase the reproducibility of STR genotyping of samples containing highly degraded DNA. Starting from a standard extraction protocol, systematic variation of individual parameters resulted in optimized protocols for three categories of ancient human bone material (different degrees of DNA degradation) and a consensus protocol for the extraction of a broad range of ancient DNA preservation states.

DNA↗
PubMed1999

Ancient DNA profiling by megaplex amplications.

Simultaneous amplification of nine human short tandem repeat (STR) DNA sequences and the amelogenin locus allows reducing to an absolute minimum the amount of sample material that is necessary for genetic identification or kinship analysis. Valuable remains can be studied this way without any visible damage, as is demonstrated by typing the DNA of a tooth root from the Saxon warrior Widukind, who died about 1200 years ago. The broad applicability of the megaplex approach is shown by typing bone and teeth specimens ranging from a few months to 3000 years of age employing AmpFISTR Profiler Plus. Additionally, megaplex STR typing is the method of choice for proving the authenticity of molecular results derived from ancient degraded DNA.

Amelogenin↗
PubMed1999

DNA profiling of orangutan (Pongo pygmaeus) feces to prove descent and identity in wildlife animals.

In this study, orangutan (Pongo pygmaeus ssp.) DNA was extracted from excrements, obtained from individual zoo populations. To examine whether human short tandem repeat loci (STR) are suitable for the reconstruction of kinship in orangutans, nine STRs, commonly used in forensic studies in man and the amelogenin system, were amplified in a multiplex polymerase chain reaction (PCR) approach. Five of the nine human autosomal STRs in question amplified successfully in orangutans. Furthermore, the experiments revealed that orangutans possess allele distributions comparable to humans in these systems. Fragment length determination allowed kinship structures to be reconstructed.

Animals↗
PubMed1999

Proving the authenticity of ancient DNA by comparative genomic hybridization.

In PCR-supported amplification of ancient, degraded DNA, contamination with contemporary DNA can lead to false-positive results, which frequently give rise to discussions in which the mere existence of ancient DNA is doubted. Our confirmation of ancient DNA using comparative genome hybridization (CGH) eliminates these doubts. Unlike PCR methods, CGH requires no amplification of the DNA to be analyzed if adequate amounts of specimen DNA is used. Thus, false results traceable to contaminations are practically ruled out. The examples provided here prove the authenticity of ancient DNA for a 250-year-old and a 3,000-year-old sample. At the same time, the CGH of ancient DNA offers the chance to gain insight into the pattern of DNA degradation and to monitor the preservation of certain chromosomal segments.

Chromosomes, Human↗
PubMed1999

Amplification of Y-chromosomal STRs from ancient skeletal material.

The adaptation to ancient DNA analysis of a Y-chromosomal STR (short tandem repeat) multiplex comprising the four STR systems DYS19, DYS390, and DYS389I/II shows the suitability of Y-chromosomal STR typing on ancient human remains. A new primer site for the system, DYS389I/II, resulting in products shortened by 94 bp, was chosen to serve the special needs of amplification of ancient DNA. For the first time, it was possible to amplify STR loci of the Y chromosome from historical and prehistorical bones of up to 3000 years old.

Bone and Bones↗
PubMed1999

Vernal keratoconjunctivitis in a Stockholm ophthalmic centre--epidemiological, functional, and immunologic investigations.

PURPOSE: To classify a cohort of 62 patients with vernal keratoconjunctivitis (VKC) in immunologic, functional, and epidemiological terms. METHODS: A retrospective chart review was conducted to establish the patients' ethnic origin and to ascertain the results of standard in vitro and in vivo testing for atopic allergy. The latter data were compared with tear Phadiatop, an allergy screening test, in 31 subjects. Further subgroup analyses included methacholine bronchial provocations, serum screening for chlamydial antibodies and epidemiological calculations of the observed prevalence of VKC in the Stockholm area in 1994. RESULTS: Thirty-seven subjects (59.7%) were sensitised to common allergens. No additional allergic subjects were diagnosed with the tear Phadiatop test. The serology for ocular chlamydial disease was negative. Only 6 out of 17 subjects displayed bronchial reactivity to methacholine of whom 4 had a history of asthma. VKC was clearly more common in individuals with an Asian and African origin. CONCLUSIONS: Sensitivity to allergens is a strong determinant for the disease but in a large proportion of the subjects this immunologic abnormality is absent. The varying prevalence of the condition in different ethnic groups indicates a genetic predisposing factor.

Adolescent↗
PubMed1999

Adrenoceptors mediating the cardiovascular and metabolic effects of alpha-methylnoradrenaline in humans.

alpha-Methylnoradrenaline is a widely used tool to study alpha2-adrenoceptor function, but its selectivity for this receptor has not been validated in humans in vivo. To characterize the adrenoceptors mediating cardiovascular and metabolic effects of alpha-methylnoradrenaline in humans, we have performed graded i.v. infusions of alpha-methylnoradrenaline in a randomized, placebo-controlled crossover study in six young, healthy males in the absence and presence of the beta-adrenoceptor antagonist propranolol, the alpha1-adrenoceptor antagonist doxazosin, and the alpha2-adrenoceptor antagonist yohimbine. alpha-Methylnoradrenaline dose-dependently increased heart rate, systolic blood pressure, cardiac output, blood glucose, serum insulin, free fatty acids, and gastrin, shortened the duration of heart rate-corrected electromechanical systole, and decreased diastolic blood pressure, total peripheral resistance, and plasma noradrenaline. Propranolol completely reversed the rise in heart rate and cardiac output, the fall in peripheral resistance, the shortening of electromechanical systole, and the rise in insulin; it blunted the increase in free fatty acids and gastrin. Yohimbine did not significantly influence most parameters but significantly potentiated the rise in insulin, blunted the increase in glucose, and prevented the fall in noradrenaline. Doxazosin was largely without effect on any of these parameters. We conclude that i.v. administered alpha-methylnoradrenaline primarily acts on beta-adrenoceptors in the human cardiovascular and metabolic system, but an alpha2-adrenergic component of the response is detectable for changes of plasma noradrenaline, blood glucose, and serum insulin. Whereas alpha-methylnoradrenaline is selective for alpha2- over alpha1-adrenoceptors, beta-adrenoceptor blockade is required to unmask alpha-adrenoceptor-mediated vasoconstriction.

Adult↗
PubMed1998

The NH2 terminus of titin spans the Z-disc: its interaction with a novel 19-kD ligand (T-cap) is required for sarcomeric integrity.

Titin is a giant elastic protein in vertebrate striated muscles with an unprecedented molecular mass of 3-4 megadaltons. Single molecules of titin extend from the Z-line to the M-line. Here, we define the molecular layout of titin within the Z-line; the most NH2-terminal 30 kD of titin is located at the periphery of the Z-line at the border of the adjacent sarcomere, whereas the subsequent 60 kD of titin spans the entire width of the Z-line. In vitro binding studies reveal that mammalian titins have at least four potential binding sites for alpha-actinin within their Z-line spanning region. Titin filaments may specify Z-line width and internal structure by varying the length of their NH2-terminal overlap and number of alpha-actinin binding sites that serve to cross-link the titin and thin filaments. Furthermore, we demonstrate that the NH2-terminal titin Ig repeats Z1 and Z2 in the periphery of the Z-line bind to a novel 19-kD protein, referred to as titin-cap. Using dominant-negative approaches in cardiac myocytes, both the titin Z1-Z2 domains and titin-cap are shown to be required for the structural integrity of sarcomeres, suggesting that their interaction is critical in titin filament-regulated sarcomeric assembly.

Actinin↗
PubMed1998

Screening for chlamydial infections and the risk of ectopic pregnancy in a county in Sweden: ecological analysis.

OBJECTIVES: To analyse trends in rates of genital chlamydial infection and ectopic pregnancy between 1985 and 1995 in a county in Sweden. SETTING: Uppsala county where screening for chlamydial infection, treatment, and contact tracing has been widespread and where ectopic pregnancies are recorded. METHODS: Rates of chlamydial infections and ectopic pregnancy between 1985 and 1995 were calculated for women aged 20-39 years. Poisson and linear regression were used to examine the association between the risk of ectopic pregnancy and the current rates of chlamydial infection and rates of chlamydial infection from up to 5 years earlier. MAIN OUTCOME MEASURES: Rates of chlamydial infection per 100 examinations, rates of ectopic pregnancy per 1000 pregnancies, rate ratios and 95% confidence intervals for an increase in chlamydial infections of 5 new cases per 100 examinations. RESULTS: 103 870 cervical samples from women aged 15-39 years were analysed; 5648 (5.4%) were positive for chlamydia. 51 630 pregnancies were analysed; 930 (1.8%) were ectopic. Both rates declined over time. For women aged 20-24 years there was a strong correlation between the rate of ectopic pregnancy and the rate of chlamydial infection in the same year (r=0.93, P<0. 001); among older women correlations were stronger with rates of chlamydial infection occurring 1 or 2 years earlier. In Poisson regression ectopic pregnancy was most strongly associated with the current rate of chlamydial infection among women aged 20-24 (rate ratio 1.85, 95% confidence interval 1.44 to 2.38), with the rate of infection 1 year earlier among 25-29 year olds (rate ratio 1.72, 1. 33 to 2.22) and 30-34 year olds (rate ratio 2.27, 1.53 to 3.37); and with the rate 2 years earlier among 35 to 39 year olds (rate ratio 2. 58, 1.45 to 4.60). CONCLUSIONS: Declining rates of genital chlamydial infections have probably led to a fall in the rate of ectopic pregnancies. The timing of the decline in the rate of ectopic pregnancies varies with age. Among young women falling rates of chlamydial infection have been accompanied by an immediate reduction in the rate of ectopic pregnancy.

Adolescent↗
PubMed1998

Comparative gene expression profiling by oligonucleotide fingerprinting.

The use of hybridisation of synthetic oligonucleotides to cDNAs under high stringency to characterise gene sequences has been demonstrated by a number of groups. We have used two cDNA libraries of 9 and 12 day mouse embryos (24 133 and 34 783 clones respectively) in a pilot study to characterise expressed genes by hybridisation with 110 hybridisation probes. We have identified 33 369 clusters of cDNA clones, that ranged in representation from 1 to 487 copies (0.7%). 737 were assigned to known rodent genes, and a further 13 845 showed significant homologies. A total of 404 clusters were identified as significantly differentially represented (P < 0.01) between the two cDNA libraries. This study demonstrates the utility of the fingerprinting approach for the generation of comparative gene expression profiles through the analysis of cDNAs derived from different biological materials.

Animals↗
PubMed1998

Genital infections among antenatal care attendees in Cape Verde.

In a cross-sectional study, 350 pregnant Capeverdian women were examined to assess the prevalence of Chlamydia trachomatis infection (CT), Neisseria gonorrhoeae infection (NG) and Bacterial vaginosis (BV). Among various analytic methods used, the polymerase chain reaction PCR (for NG, CT) yielded a higher detection rate than did direct microscopy or culture (NG), or direct immuno-fluorescence (CT). Since the PCR analytic of air-dried specimens is not hampered by harsh storage and transport conditions, it could serve to validate other detection methods where laboratory facilities are suboptimal. Among sociodemographic risk factors young age, and currently living alone, were significantly associated with infection.

Adult↗
PubMed1997

Comparison of the Ca2+-binding properties of human recombinant calretinin-22k and calretinin.

Calretinin-22k (CR-22k) is a splice product of calretinin (CR) found specifically in cancer cells, and possesses four EF-hands and a differently processed C-terminal end. The Ca2+-binding properties of recombinant human calretinin CR-22k were investigated by flow dialysis and spectroscopic methods and compared with those of CR. CR possesses four Ca2+-binding sites with positive cooperativity (nH = 1.3) and a [Ca2+]0.5 of 1.5 microM, plus one low affinity site with an intrinsic dissociation constant (K'D) of 0.5 mM. CR-22k contains three Ca2+-binding sites with nH of 1.3 and [Ca2+]0.5 of 1.2 microM, plus a low affinity site with K'D of 1 mM. All the sites seem to be of the Ca2+-specific type. Limited proteolysis and thiol reactivity suggest that that the C terminus of full-length CR, but not of CR-22k, is in close proximity of site I leading to mutual shielding. Circular dichroism (CD) spectra predict that the content of alpha-helix in CR and CR-22k is similar and that Ca2+ binding leads to very small changes in the CD spectra of both proteins. The optical properties are very similar for CR-22k and CR, even though CR-22k possesses one additional Trp at the C-terminal end, and revealed that the Trp residues are organized into a hydrophobic core in the metal-free proteins and become even better shielded from the aqueous environment upon binding of Ca2+. The fluorescence of the hydrophobic probe 2-p-toluidinylnaphtalene-6-sulfonate is markedly enhanced by the two proteins already in the absence of Ca2+ and is further increased by binding of Ca2+. The trypsinolysis patterns of CR and CR-22k are markedly dependent on the presence or absence of Ca2+. Together, our data suggest the presence of an allosteric conformational unit encompassing sites I-III for CR-22k and I-IV for CR, with a very similar conformation and conformational changes for both proteins. In the allosteric unit of CR, site IV is fully active, whereas in CR-22k this site has a 80-fold decreased affinity, due to the decreased amphiphilic properties of the C-terminal helix of this site. Some very specific Ca2+-dependent conformational changes suggest that both CR and CR-22k belong to the "sensor"-type family of Ca2+-binding proteins.

Calbindin 2↗
PubMed1997

[Molecular sex determination in skeletal remains of premature and newborn infants of the Aegerten, Switzerland, burial field].

The morphological and morphometrical analyses of skeletal remains usually give reliable access to the gender of mature individuals while the analyses of skeletal remains of immature individuals allocate only 70-90% of the individuals. However, the use of modern techniques like the Polymerase Chain Reaction (PCR) enables a sex identification also if fragmentary material or skeletal remains of children are investigated. The present study reconstructs the sex ratio of about 120 stillborn and neonate individuals of the burial place Aegerten, Switzerland. The morphometrical sex determination of the children suggests a large excess (about 60%) of female individuals. This finding was compared to the molecular sex identification. In order to perform a molecular sex identification aDNA was extracted from bone samples of the stillborn and neonate individuals. A standard phenol/chloroform extraction and a purification with a silica powder were carried out. Finally, the aDNA samples were amplified with a primer system for the amelogenin gene, that is located on the human sex chromosomes.

Bone and Bones↗
PubMed1997

[Detection of DNA single-copy sequences of prehistoric teeth. Site milieu as a factor for preservation of DNA].

DNA-extracts from prehistoric roots of teeth were analyzed by PCR to investigate microsatellite-systems. At the same time the x/y-chromosome specific system Amel A/B was investigated. The samples were collected from different burial conditions of similar age. Factors leading to limited DNA-degradation are given. For the first time reproducible Quadruplex-amplification on single-copy loci of prehistoric tissue was obtained in samples from the Lichtenstein-Cave, Kr. Osterode/Harz. Positive results are explained by low temperature in the burial site. In addition the results show that desiccation allows DNA-preservation but cannot protect the DNA from damage due to microbial origin. Microorganisms can destroy DNA-structures completely.

Burial↗