Simultaneous detection and typing of influenza viruses A and B by a nested reverse transcription-PCR: comparison to virus isolation and antigen detection by immunofluorescence and optical immunoassay (FLU OIA).

A nested reverse transcription (RT)-PCR was developed for simultaneous detection and typing of influenza viruses A and B. The detection limit for influenza virus A subtypes H1 and H3 and that for influenza virus B were between 1 and 4 target gene copies per reaction for each type. The clinical benefit of the RT-PCR method was evaluated by comparing the results with virus isolation and direct immunofluorescence (IF) assays on 215 nasopharyngeal aspirates from patients with suspected influenza virus infection.

Large-scale screen for genes controlling mammalian embryogenesis, using high-throughput gene expression analysis in mouse embryos.

We have adapted the whole-mount in situ hybridization technique to perform high-throughput gene expression analysis in mouse embryos. A large-scale screen for genes showing specific expression patterns in the mid-gestation embryo was carried out, and a large number of genes controlling development were isolated. From 35760 clones of a 9.

Large-scale screen for genes involved in gonad development.

The vertebrate gonad develops from the intermediate mesoderm as an initially bipotential organ anlage, the genital ridge. In mammals, Sry acts as a genetic switch towards testis development. Sox9 has been shown to act downstream of Sry in testis development, while Dax1 appears to counteract Sry.

Chlamydophila abortus in a Brown skua (Catharacta antarctica lonnbergi) from a subantarctic island.

On Bird Island, South Georgia, a new strain of Chlamydophila abortus was detected in one Brown skua out of 37 specimens from six different seabird species. Phylogenetic analysis of the rnpB and omp1 genes indicated the strain to be more closely related to C. abortus than to 6BC, the type strain of Chlamydophila psittaci.

Characterization of the rnpB gene and RNase P RNA in the order Chlamydiales.

The sequence of the RNase P RNA gene (rnpB) was determined for 60 strains representing all nine species in the family Chlamydiaceae and for the related Chlamydiales species, Parachlamydia acanthamoebae and Simkania negevensis. These sequences were used to infer evolutionary relationships among the Chlamydiaceae. The analysis separated Chlamydophila and Chlamydia into two lineages, with Chlamydophila forming three distinct clusters: the Chlamydophila pneumoniae strains; the Chlamydophila pecorum strains; and a third cluster comprising the species Chlamydophila psittaci, Chlamydophila abortus, Chlamydophila caviae and Chlamydophila felis.

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.

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.

Palaeogenetic analysis of (pre)historic artifacts and its significance for anthropology.

The possibility of isolating ancient DNA (aDNA) from all kinds of (pre)historic anthropogenetic artifacts opens new perspectives. This study applies palaeogenetic techniques to three anthropological issues: 1. Palaeodiet.

Reconstruction of kinship by fecal DNA analysis of orangutans.

Genetic analysis is a useful tool for assigning biological relationships. Thus, it will improve genetic management of wild animal populations and breeding colonies. Kinship analysis will give new insights into the behavior, sociobiology and genetic management of orangutans.

STR allelic frequencies in a German skeleton collection.

Chromosomal DNA was isolated from bones from a German skeleton collection (Goslar, 18th century) and detected by PCR. Nine microsatellite regions were amplified by multiplex reactions using the AmpFlSTR Profiler Plus kit and analysed to obtain their allelic distribution. A statistical evaluation of the results revealed no allelic differentiation between the historic sample and a modern German one at each locus.

Ancient DNA-typing approaches for the determination of kinship in a disturbed collective burial site.

Several DNA-typing approaches are applied for identification and kinship analysis. Autosomal Short Tandem Repeat (STR) typing produces the genetic fingerprint that is unique to an individual. Y-chromosomal STR typing identifies individuals of the same paternal lineage, and sequence analysis of the hypervariable region of the mitochondrion can identify maternally related individuals.

STR-genotyping of archaeological human bone: experimental design to improve reproducibility by optimisation of DNA extraction.

The analysis of degraded DNA with the help of short tandem repeat loci (STRs) is an important source of information both in forensic casework and in the anthropological context. The reproducibility of STR-genotyping of highly degraded or "ancient" DNA can be reduced by the generation of artifacts during PCR amplification. The frequency and amount of these artifacts--allelic dropout and the generation of shadow bands--are related to the quality and quantity of the extracted DNA amplified in a PCR reaction.

Megaplex DNA typing can provide a strong indication of the authenticity of ancient DNA amplifications by clearly recognizing any possible type of modern contamination.

Recent experiments revealed the perfect applicability of megaplex typing by autosomal short tandem repeats (STRs) to degraded DNA. The advantages of megaplex approaches lie in reduced amounts of sample material that are necessary and in remarkable time saving. Furthermore, megaplex typing clearly recognizes possible contaminations and thus has a large potential for indicating authenticity in ancient DNA analysis.

Evaluation of morphological sex determinations by molecular analyses.

The study presents an evaluation of morphological sex determinations of adult skeletal individuals based on traits of the ossa coxae and the cranium. The evaluation criterion was genetic analysis of the amelogenine gene, which represents parts of the X- and the Y-chromosome (Mannucci et al. 1994).

Molecular sex identification of stillborn and neonate individuals ("Traufkinder") from the burial site Aegerten.

The study reconstructs the sex ratio of 121 stillborn and neonate individuals from the early modern burial site Aegerten, Switzerland. The immature individuals, who were not baptised before death, were buried along the walls of the church. To perform a molecular sex identification, bone samples from the infants were collected from different skeletal elements.

The paired homeobox gene Uncx4.1 specifies pedicles, transverse processes and proximal ribs of the vertebral column.

The axial skeleton develops from the sclerotome, a mesenchymal cell mass derived from the ventral halves of the somites, segmentally repeated units located on either side of the neural tube. Cells from the medial part of the sclerotome form the axial perichondral tube, which gives rise to vertebral bodies and intervertebral discs; the lateral regions of the sclerotome will form the vertebral arches and ribs. Mesenchymal sclerotome cells condense and differentiate into chondrocytes to form a cartilaginous pre-skeleton that is later replaced by bone tissue.

Brachyury is a target gene of the Wnt/beta-catenin signaling pathway.

To identify target genes of the Wnt/beta-catenin signaling pathway in early mouse embryonic development we have established a co-culture system consisting of NIH3T3 fibroblasts expressing different Wnts as feeder layer cells and embryonic stem (ES) cells expressing a green fluorescent protein (GFP) reporter gene transcriptionally regulated by the TCF/beta-catenin complex. ES cells specifically respond to Wnt signal as monitored by GFP expression. In GFP-positive ES cells we observe expression of Brachyury.

A protein kinase encoded by the t complex responder gene causes non-mendelian inheritance.

Males heterozygous for the t-haplotype form of mouse chromosome 17 preferentially transmit the t-chromosome to their progeny. Several distorter/sterility loci carried on the t-haplotype together impair flagellar function in all spermatozoa whereas the responder, Tcr, rescues t-sperm but not wild-type sperm. Thus, t-sperm have an advantage over wild-type sperm in fertilizing egg cells.

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.

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.

Kidney-specific cadherin (cdh16) is expressed in embryonic kidney, lung, and sex ducts.

Cdh16 was initially described as a truncated cadherin expressed in the adult rabbit kidney. We have analyzed the expression pattern of cdh-16 during mouse embryogenesis, and show that cdh-16 transcripts are present in ureter-derived epithelia of the metanephric kidney. In addition, we demonstrate that cdh-16 is also transiently expressed in the epithelia of embryonic sex ducts and the lung of the embryo.

Diurnal changes in xylem pressure of the hydrated resurrection plant Myrothamnus flabellifolia: evidence for lipid bodies in conducting xylem vessels.

The resurrection plant Myrothamnus flabellifolia has the ability to recover from repeated prolonged and extreme desiccation cycles. During the dry state the inner walls of the xylem vessels seemed to be covered, at least partly, by a lipid film as shown by Sudan III and Nile Red staining. The lipid film apparently functioned as an 'internal cuticle' which prevented the adjacent parenchyma ray cells from complete water loss.

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.

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.