BACKGROUND: The serpin-enzyme complex receptor (SECR) has previously been successfully targeted for gene delivery using synthetic peptide ligands covalently linked in fluid phase to commercially available polylysine preparations (approximately 10-54kDa). The objective of the present study was to improve this approach by the use of small, bifunctional, and easily standardised synthetic peptides. METHODS: Two synthetic peptides designated polylysine antitrypsin 1 (PAT1) (K16 FNKPFVFLI) and PAT2 (K16 CSIPPEVKFNKPFVFLI) were evaluated for gene delivery to the HUH7 human hepatocyte cell line. The K16 moiety binds DNA electrostatically, while the FVFLM motif of human alpha1-antitrypsin targets the SECR. RESULTS: Both PAT1 and PAT2 bind to and condense DNA into small particles as shown by laser scattering techniques. However, only PAT2 is effective for gene delivery, presumably on account of the greater distance between the K16 chain and the FVFLM motif. Gene delivery by PAT2/DNA complexes is chloroquine-dependent, can be blocked completely by free ligand (CSIPPEVKFNKPFVFLI), and is highly efficient (e.g. approximately five-fold more effective than lipofectamine). At physiological salt concentrations, PAT2/DNA complexes formed at 4 microg/ml DNA are approximately 350 nm in diameter and highly effective for gene transfer, but at 100 microg/ml DNA the complexes are aggregated (diameter > 4 microm) and inactive. CONCLUSIONS: A small (33 amino acid), bifunctional, synthetic peptide represents a highly efficient and readily standardised DNA vector for the SECR. The effectiveness of this peptide depends on the distance of the K16 moiety from the targeting ligand. High salt concentrations are not required to form effective vector/DNA complexes.

Carcinoma, Hepatocellular↗

PubMedDate not supplied

Correlation between high-field T2-weighted MR imaging and histology of ischemic lesions in gerbil brain.

Global forebrain ischemia in the Mongolian gerbil is a common animal model for use in stroke research. We produced lesions of graded severity in gerbil brains (after prescreening by MR imaging) by performing 6-minute bilateral carotid artery occlusions while monitoring pericranial temperature with a temporalis muscle thermocouple probe and maintaining the temperature at 32 degrees C, 36 degrees C, or 40 degrees C. Lesion severity was scored 4 days after occlusion from findings on spin-echo images acquired at 7 T and from histologic scores. Statistically significant correlation was observed between the MR imaging score and brain temperature and between the MR imaging score and the area of the CA region of the hippocampus measured by histology. In addition, because prescreening with MR imaging revealed abnormalities in the hippocampus of some of the animals, and these animals were rejected from the study, the statistical significance of the result could be strengthened.

Animals↗

PubMedDate not supplied

Diagnosis of intrathoracic tuberculosis by detection of tubercle and/or tubercular antigen (TB Ag) in bronchial aspirate.

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Antigens, Bacterial↗

PubMedDate not supplied

"Myocardial infarction" in adolescents: do we have the correct diagnosis?

The evaluation of adolescents with chest pain, elevated cardiac enzymes, and abnormal electrocardiograms (ECGs) continues to pose diagnostic and management dilemmas. Myocardial infarction is an uncommon finding in this population and alternative diagnoses must be considered. Our database was retrospectively reviewed for adolescents age 16-18 years without prior cardiac history who underwent cardiac catheterization. Patients who presented with chest pain, elevated cardiac enzymes, normal ejection fraction, and abnormal ECGs were included. Management, diagnostic testing, and final diagnosis were reviewed. Nine adolescents (eight males and one female) without prior cardiac history were identified. The ECG findings in all patients were consistent with myocardial ischemia in a coronary distribution. Thrombotic coronary occlusion was not found in any patient. In adolescents without prior cardiac history of risk factors for myocardial infarction such as Kawasaki disease, familial hypercholesterolemia, or drug use who present with chest pain, multiple diagnoses must be considered even in the presence of focal ischemic ECG changes and elevated cardiac enzymes. Thrombolytic therapy or anticoagulation should be withheld until a definitive diagnosis of myocardial infarction has been made. Magnetic resonance imaging is the most useful tool to differentiate focal myocarditis from myocardial infarction.

Adolescent↗