Plasma levels of adenosine 3',5'-monophosphate in the fetoplacental unit and maternal circulation at delivery

Adenosine 3',5'-monophosphate (cAMP) was measured in matched samples of maternal plasma (MV) and umbilical artery (UA) and vein (UV) of 14 normal infants born following an uncomplicated vaginal delivery. There was good correlation between UA and UV cAMP levels, which seems to indicate that the fetoplacental unit maintains a constant equilibrium of the nucleotide concentration in the umbilical circulation. There was a markedly increased UA cAMP concentration at term relative to that of MV, as well as an arteriovenous gradient of plasma cAMP level (UA greater than UV) in the fetoplacental circulation, which strongly suggest that the high cAMP levels in the cord blood originate mainly from the fetus.     

Type-specific efficacy of acellular pertussis vaccine

Two types of acellular pertussis vaccine have been used in Japan since their introduction in 1981. They are different in their antigen content and immunogenicity. To assess the type-specific efficacy of acellular pertussis vaccine commercially available in Japan, 442 households of patients with pertussis were surveyed from 1981 to 1987. The secondary attack rates in children 2 through 8 years of age were 61.3% (46/75) in unimmunized children, 7.5% (4/53) in children fully immunized by the filamentous hemagglutinin predominant type acellular pertussis vaccine, 14.3% (1/7) in children fully immunized by the pertussis toxin-filamentous hemagglutinin type vaccine, and 13.5% (7/52) in children given whole-cell pertussis vaccine. The estimated efficacy was 87.7% for the filamentous hemagglutinin predominant type vaccine and 76.7% for the pertussis toxin-filamentous hemagglutinin type vaccine in children aged 2 through 8 years. Both types of acellular pertussis vaccine were similarly effective, without any statistically significant differences, and their efficacy was not different from that of whole-cell vaccines. This survey also indicated that adults are a major source of infection, with a high secondary attack rate of 7.8%.     

Aerosolization of lipoplexes using AERx Pulmonary Delivery System

The lung represents an attractive target for delivering gene therapy to achieve local and potentially systemic delivery of gene products. The objective of this study was to evaluate the feasibility of the AERx Pulmonary Delivery System for delivering nonviral gene therapy formulations to the lung. We found that "naked" DNA undergoes degradation following aerosolization through the AERx nozzle system. However, DNA formulated with a molar excess of cationic lipids (lipoplexes) showed no loss of integrity. In addition, the lipoplexes showed no significant change in particle size, zeta (zeta) potential, or degree of complexation following extrusion. The data suggest that complexation with cationic lipids had a protective effect on the formulation following extrusion. In addition, there was no significant change in the potency of the formulation as determined by a transfection study in A-549 cells in culture. We also found that DNA formulations prepared in lactose were aerosolized poorly. Significant improvements in aerosolization efficiency were seen when electrolytes such as NaCl were added to the formulation. In conclusion, the data suggest that delivery of lipoplexes using the AERx Pulmonary Delivery System may be a viable approach for pulmonary gene therapy.     

Performance characteristics of updated INNO-LiPA assays for molecular typing of human leukocyte antigen A (HLA-A), HLA-B, and HLA-DQB1 alleles

We carried out a multicenter performance evaluation of three new DNA-based human leukocyte antigen (HLA) typing assays: INNO-LiPA HLA-A Update, INNO-LiPA HLA-B Update, and INNO-LiPA HLA-DQB1 Update. After optimization, the accuracy rates were all 100%, and the final observed resolutions were 99.4, 92.4, and 85.6%, respectively. These rapid and easy-to-perform assays yielded results fully concordant with other DNA-based tissue typing tests.     

Functional cross-talk between cytokine receptors revealed by activating mutations in the extracellular domain of the beta-subunit of the GM-CSF receptor

Several reports have suggested an interaction between the erythropoietin receptor (EpoR) and the shared signaling subunit (hbeta(c)) of the human granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-3, and IL-5 receptors, although the functional consequences of this interaction are unclear. We previously showed that in vivo expression of constitutively active extracellular (EC) mutants of hbeta(c) induces erythrocytosis and Epo independence of erythroid colony-forming units (CFU-E). This occurs despite an apparent requirement of these mutants for the GM-CSF receptor alpha-subunit (GMRalpha), which is not expressed in CFU-E. Here, we show that coexpression of hbeta(c) EC mutants and EpoR in BaF-B03 cells, which lack GMRalpha, results in factor-independent proliferation and JAK2 activation. Mutant receptors that cannot activate JAK2 fail to produce a functional interaction. As there is no detectable phosphorylation of hbeta(c) on intracellular tyrosine residues, EpoR displays constitutive tyrosine phosphorylation. These observations suggest that JAK2 activation mediates cross-talk between EC mutants of hbeta(c) and EpoR. The implications of these data are discussed as are our findings that activated hbeta(c) mutants can functionally interact with certain other cytokine receptors.