Intraamniotic administration of an adenoviral vector for gene transfer to fetal sheep and mouse tissues

Replication-deficient adenoviruses have been used to transfer various genes of interest to mammalian tissues in vivo. Effective gene therapy for inborn genetic defects presenting with significant morbidity and mortality at birth will require correction of the defect prenatally. To test the hypothesis that intra-amniotically administered adenovirus transfers gene expression to fetal tissues, replication-deficient human type 5 adenovirus carrying the lacZ gene which encodes nuclear-targeted bacterial beta-galactosidase (Av1LacZ4) was instilled into the amniotic cavity of fetal sheep (10(10) to 1.5 x 10(11) pfu) and fetal mice (10(9) pfu) at 0.8 term gestation. Amniotic membranes and gastrointestinal and respiratory tract tissues were harvested after 3 d, bacterial beta-galactosidase activity was determined by 5-bromo-4-chloro-3-indoyl-beta-D-galactopyranoside (X-gal) enzyme-histochemistry, and tissue integrity was assessed in sections stained with hematoxylin and eosin. Bacterial beta-galactosidase activity was abundant in amniotic membranes and present in lower levels in esophagus, stomach, and small intestine as well as in conducting airways and pulmonary alveoli. To determine whether gene transfer by intraamniotic injection of adenovirus was dose-dependent, Av1Luc1, an adenoviral vector carrying the gene for luciferase (10(5)-10(9) pfu), was injected intraamniotically into fetal mice at 0.8 term gestation. Luciferase activity measured after 3 d in tissue homogenates of Av1Luc1-treated fetal mice revealed a linear dose response in amniotic membranes and gastrointestinal and respiratory tract organs. Intraamniotic administration of an adenoviral gene vector leads to expression of the transferred gene in amniotic membranes as well as in fetal gastrointestinal and respiratory tract tissues in a dose-dependent manner.     

Total pancreatectomy for carcinoma of the pancreas

Phacoanaphylaxis (lens-induced uveitis) is a potentially curable ocular inflammation secondary to lens truma. Recently we have seen three clinically undiagnosed enucleation specimens in our eye pathology laboratory with this entity. Our observations add an argument to the recent concept that early lens removal in cases of traumatic cataract with lens capsule rupture would ultimately lead to better visual results.     

Temperature-sensitive DNA polymerase induced by a bacteriophage T5 mutant: relationship between polymerase and exonuclease activities

DNA polymerase induced by bacteriophage T5ts53, a mutant with temperature-sensitive polymerase, was purified to about 95% purity as judged by dodecyl sulfate gel electrophoresis. The 3' leads to 5' exonuclease associated with the polymerase had higher activity than that associated with the parent wild-type enzyme. It was more stable to heat than the polymerase, and it degraded primer-template even in the presence of 4 dNTP's at higher temperature. However, the evidence presented shows that the inhibition of DNA synthesis by higher temperature was primarily due to defects in polymerase function rather than to overactive exonuclease. The presence of primer-template DNA stabilized the polymerase to heat. Purified ts53 polymerase was also shown to discriminate against incorportion of BrdUMP, especially at higher temperature. This is an agreement with observations made in vivo with ts53-infected bacteria.     

Patient responses to request for written permission to review medical records

2,163 patients of 55 physicians in one county, were sent letters asking for their written permission to have their medical records audited by the University of Michigan research team. The overall positive response rate was 64.1% for one county. Differences in response rates were statistically significant among specialties, among diagnoses, and among individual physicians.     

System C: an essay in human relatedness

The system of dyadic human relationship, designated System C, is examined in terms of unique qualities and measures useful for both research and teaching in the area of dynamic psychiatric interviews. Implications are indicated in polyadic systems, and a new aspect of transference is noted. A sample of interview material is analyzed as illustration.