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Biomedical subjects

L Ernst

Publications and source records attributed to L Ernst.

12 recordsLinked to original sources
PubMed1992

Sensitive detection of RNAs in single cells by flow cytometry.

A rapid and sensitive fluorescent in situ hybridization method has been developed to probe RNA contents of individual cells by flow cytometry. Fixed cells in suspension were hybridized with 5' end-fluorophore-labeled oligodeoxynucleotides complementary to defined regions of the RNA of interest and analyzed by flow cytometry. With this method, we monitored combinations of histone H4 mRNA, 18S rRNA and 28S rRNA levels in synchronized HeLa S3 cells by multicolor analysis. A fluorescence signal equivalent to 1800 copies of histone H4 mRNA per cell was detected with signal-to-background ratio of 5.4. If non-specific binding of the fluorophore-labeled probe can be reduced, as few as 100 copies of mRNA of the size of H4 could be detected in individual cells by flow cytometry.

Base Sequence↗
PubMed1992

Specific and sensitive measurement of FK506 and its metabolites in blood and urine of liver-graft recipients.

A specific and sensitive assay for quantifying the immunosuppressant FK506 and its metabolites in blood and urine was developed. 32-O-Acetyl FK506 was synthesized and used as internal standard. FK506 and its metabolites were purified from the samples by solid-liquid extraction and were injected into a high-performance liquid chromatographic (HPLC) system linked to a mass spectrometer (MS) by particle-beam interface. The FK506 derivatives were separated from interfering material by use of a 100 x 4 mm C8 analytical column and water/acetonitrile or water/methanol gradient elution; they were detected by negative chemical ionization with methane as reagent gas. The limit of detection was 25 pg in a standard solution, and the limit of quantification in blood was 250 pg (extracted from 1 mL of blood). The CV was 11.3% at 5 ng, and no interferences with other drugs were found.

Chromatography, High Pressure Liquid↗
PubMed1986

A unique immune response control locus mapping with the H-2D class I region.

The immune response to the polysaccharide antigen Trinitrophenyl (TNP)-ficoll is controlled by two complementing loci in the murine major histocompatibility complex. One locus of control is present in the I-A subregion and the second is located between the S and D regions. A monoclonal antibody 48-21.7 was selected for its ability to significantly block the in vitro primary response to TNP-ficoll. This antibody bound to antigen-presenting cells does not interfere with the presentation of other antigens. Experiments are presented to demonstrate that the monoclonal antibody 48-21.7 precipitates a unique protein of approximately 40,000 daltons. Sequential immunoprecipitation with monoclonal antibodies to the class I Db antigen and to relevant Ia antigens demonstrated no cross-reactivity. Amino acid sequencing has found unique amino acids to be present at three class I N-terminal invariant positions. These observations indicate the presence of a unique class I-like molecule coded for by genes mapping between S and D and involved in the response to the polysaccharide ficoll.

Animals↗
PubMed1978

The psychological effects of sudden infant death syndrome on surviving family members.

It has been estimated that 7,000 to 10,000 deaths each year may be attributed to Sudden Infant Death Syndrome (SIDS). Although some clinicians have published their personal observations regarding the psychological effects of SIDS on surviving family members, a literature search failed to reveal more thorough studies of the aftermath of SIDS. This study reports the responses to a 13-page questionnaire of 32 parents who had experienced SIDS. Several major findings were observed: (1) SIDS is the most severe crisis these parents had ever experienced, taking their families an average of 8.3 months to regain the level of family organization they had held prior to the death, and taking individual parents an average of 15.9 months to regain the level of personal happiness they had held prior to the death; (2) a majority of parents suffered personal guilt, and numerous other psychological and/or physiological difficulties; (3) relationships with other family members were affected in various ways in the vast majority of cases; and (4) 60 percent of parents who had experienced SIDS in this particular population could not be found for participation in the study; all of these parents had, within 2 1/2 years of the death, moved from their home towns. Implications for crisis prevention and intervention are outlined.

Crisis Intervention↗