ABSTRACT
We describe a rapid technique to determine numerical abnormalities of chromosomes that can be applied to slides prepared from fresh, uncultured blood samples. Primed in situ synthesis is a method that has previously been utilized as a rapid alternative to conventional fluorescence in situ hybridization for localizing repeated DNA sequences on metaphase chromosomes and interphase nuclei prepared from cultured lymphocytes. By applying the technique to uncultured preparations from fresh blood, aneuploidy analysis can be completed in less than 3 h from the collection of the blood sample.
Subject(s)
Aneuploidy , DNA/blood , Karyotyping/methods , Chromosome Aberrations , Chromosomes, Human, Pair 1 , Chromosomes, Human, Pair 7 , Chromosomes, Human, Pair 9 , DNA Primers , Humans , Lymphocytes/ultrastructure , Male , Microscopy, Fluorescence , Nucleic Acid Hybridization , Repetitive Sequences, Nucleic Acid , Time Factors , Y ChromosomeABSTRACT
We studied G-CSF concentrations ([G-CSF]) at birth and their relationship with neutrophil count, incidence of infection, gestational age, labour, and the presence of maternal pregnancy-induced hypertension. Plasma [G-CSF] were significantly elevated in babies with suspected infection and in those of hypertensive mothers, compared to healthy babies delivered by elective caesarian section (median [range] = 3101 [75- > 5000] pg/ml and 153 [45-857] pg/ml versus 32 [11-266] pg/ml; P < 0.0001); and were unrelated to neutrophil count and gestational age. Initial high concentrations (> 100 pg/ml) declined by 7 d (P < 0.0001).
Subject(s)
Granulocyte Colony-Stimulating Factor/blood , Hypertension/blood , Infant, Newborn/blood , Infections/blood , Pregnancy Complications, Cardiovascular/blood , Aging/blood , Cesarean Section , Female , Humans , Maternal-Fetal Exchange/physiology , Pregnancy , Pregnancy Complications, Infectious/bloodABSTRACT
The fusion glycoprotein of the Edinburgh strain of respiratory syncytial (RS) virus was cloned from infected cell mRNA. A full-length clone was subjected to sequence analysis, and compared with other strains of RS virus. The inferred primary amino acid sequence was used to generate a nested set of overlapping peptides spanning the mature protein. Peptides were synthesized on polyethylene pins and examined for their reactivity towards high titre human antisera. Decameric peptides spanning the highly conserved region between amino acids Lys and Ala (positions 470 to 490), reacted strongly with the sera and a detailed study with hexameric peptides located the epitope to FPSDEF, at positions 483 to 488. Replacement synthesis analysis revealed that Pro at position 484 and Glu at 487 were critical components of the binding site and could not be substituted.
