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Biomedical subjects

M Leavitt

Publications and source records attributed to M Leavitt.

14 recordsLinked to original sources

Increased titer of recombinant AAV vectors by gene transfer with adenovirus coupled to DNA-polylysine complexes.

One of the principal problems with using recombinant adeno-associated virus vectors (rAAV) as vehicles for gene delivery has been the difficulty in obtaining high-titer virus stocks after the initial transfection into producer cells. In this report we describe a method for transfecting cells at extremely high efficiency with rAAV vector DNA and complementation plasmid while simultaneously infecting those cells with replication competent adenovirus using adenovirus-polylysine-DNA complexes. We further show that this technique results in an increase in rAAV transducing titer by two orders of magnitude over what is typically achieved by standard calcium phosphate transfection.

Adenoviridae

An animal model for delirium.

This study describes an animal model for delirium comparing rats treated with either saline or atropine. The model was defined by recordings of cortical EEGs, maze performance, and behavioral observations. EEG slowing and increased amplitude, difficulty with attention and memory, sleep-wake cycle reversal, and changes in behavior (lack of focused direction, irritability, fluctuating levels of activity, excessive random sniffing) appeared consistent with signs and symptoms seen in human delirium. EEG abnormalities in atropine-treated rats returned to normal before cognitive deficits did. Motor activity monitoring did not reveal diminished motor activity as a confounding variable in maze performance.

Alcohol Drinking

Acute reduction in functional infarct expansion with late coronary reperfusion: assessment with quantitative two-dimensional echocardiography.

Reperfusion performed too late to salvage myocardium decreases chronic infarct expansion in experimental animals. However, the acute effects of delayed reperfusion are not known. Twenty-two dogs underwent 3 (n = 8), 4 (n = 8) or 6 h (n = 6) of circumflex artery occlusion followed by 3 h of reperfusion. Effects of reperfusion on diastolic expansion were assessed in two ways: 1) change in mean radius of curvature of the infarct segment, and 2) change in the ratio of the length of the diameter from the center of the infarct zone to the opposite wall (septal-lateral diameter) to the length of the diameter perpendicular to this (anteroposterior diameter). Effects on systolic expansion were examined with quantitative two-dimensional echocardiographic systolic thickening analysis. Delayed reperfusion produced an immediate decrease in diastolic infarct expansion. The ratio of septal-lateral/anteroposterior diameters, which had increased with occlusion from a preocclusion baseline of 0.98 +/- 0.06 to 1.13 +/- 0.08 (p less than 0.001), decreased with reperfusion to 1.02 +/- 0.07 at 15 min and 1.03 +/- 0.08 at 3 h of reperfusion (p = 0.001). This was due solely to a decrease in the septal-lateral diameter. The radius of curvature of the infarcted segment increased from 2.1 +/- 0.5 cm before reperfusion to 2.74 +/- 0.8 cm at 15 min and 2.6 +/- 0.85 cm at 3 h of reperfusion (p = 0.009). This occurred despite a significant (13.6%) decline in end-diastolic cavity area and is compatible with flattening of the reperfused infarct region. Systolic infarct expansion also improved slightly.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals

Superiority of quantitative exercise thallium-201 variables in determining long-term prognosis in ambulatory patients with chest pain: a comparison with cardiac catheterization.

The purpose of this study was to determine the prognostic utility of quantitative exercise thallium-201 imaging and compare it with that of cardiac catheterization in ambulatory patients. Accordingly, long-term (4 to 9 years) follow-up was obtained in 293 patients who underwent both tests for the evaluation of chest pain: 89 had undergone coronary artery bypass graft surgery within 3 months of testing and were excluded from analysis, 119 experienced no cardiac events and 91 had an event (death in 20, nonfatal myocardial infarction in 21 and coronary artery bypass operations performed greater than 3 months after cardiac catheterization in 50). When all variables were analyzed using Cox regression analysis, the quantitatively assessed lung/heart ratio of thallium-201 activity was the most important predictor of a future cardiac event (chi 2 = 40.21). Other significant predictors were the number of diseased vessels (chi 2 = 17.11), patient gender (chi 2 = 9.43) and change in heart rate from rest to exercise (chi 2 = 4.19). Whereas the number of diseased vessels was an important independent predictor of cardiac events, it did not add significantly to the overall ability of the exercise thallium-201 test to predict events. Furthermore, information obtained from thallium-201 imaging alone was marginally superior to that obtained from cardiac catheterization alone (p = 0.04) and significantly superior to that obtained from exercise testing alone (p = 0.02) in determining the occurrence of events. In addition, unlike the exercise thallium-201 test, which could predict the occurrence of all categories of events, catheterization data were not able to predict the occurrence of nonfatal myocardial infarction. The exclusion of bypass surgery and previous myocardial infarction did not alter the results. In conclusion, data from this study demonstrate that exercise thallium-201 imaging may be superior to data from both exercise testing alone and cardiac catheterization data alone for predicting future events in ambulatory patients who have undergone both exercise thallium-201 imaging and catheterization for the evaluation of chest pain.

Adult

Improving recovery following cardiac surgery: a randomized clinical trial.

To enhance individual and family health during recovery from heart surgery, this study employed nursing interventions based on self-efficacy and family stress theory during the hospitalization period and for 3 months thereafter. The effectiveness of the interventions were assessed through a randomized trial in which 67 prospective bypass and valve surgery patients, aged 30-77 years, and their spouses, were allocated either the experimental interventions or usual care and followed for 6 months. At 3 months post-surgery, the only statistically significant differences between the experimentals and controls were on perceived self-efficacy for lifting and tolerating emotional distress. At 6 months no significant differences were found on individual or family measures. Analyses revealed that age, gender and preoperative cardiac status significantly affected individual recovery. The study is continuing with a larger sample in order to explicate the recovery process and to better determine whether a low intensity nursing intervention can effect changes in individual and family recovery.

Adult

Transposon-induced symbiotic mutants of Bradyrhizobium japonicum: isolation of two gene regions essential for nodulation.

Two strains of the soybean endosymbiont Bradyrhizobium japonicum, USDA 110 and 61 A101 C, were mutagenized with transposon Tn5. After plant infection tests of a total of 6,926 kanamycin and streptomycin resistant transconjugants, 25 mutants were identified that are defective in nodule formation (Nod-) or nitrogen fixation (Fix-). Seven Nod- mutants were isolated from strain USDA110 and from strain 61 A101 C, 4 Nod- mutants and 14 Fix- mutants were identified. Subsequent auxotrophic tests on these symbiotically defective mutants identified 4 His- Nod- mutants of USDA110. Genomic Southern analysis of the 25 mutants revealed that each of them carried a single copy of Tn5 integrated in the genome. Three 61 A101 C Fix- mutants were found to have vector DNA co-integrated along with Tn5 in the genome. Two independent DNA regions flanking Tn5 were cloned from the three non-auxotrophic Nod- mutants and one His-Nod- mutant of USDA110. Homogenotization of the cloned fragments into wild-type strain USDA110 and subsequent nodulation assay of the resulting homogenotes confirmed that the Tn5 insertion was responsible for the Nod- phenotype. Partial EcoR1 restriction enzyme maps around the Tn5 insertion sites were generated. Hybridization of these cloned regions to the previously cloned nod regions of R. meliloti and nif and nod regions of B. japonicum USDA110 showed no homology, suggesting that these regions represent new symbiotic clusters of B. japonicum.

DNA Transposable Elements

Cardiac function: evaluation with fast-echo MR imaging.

Magnetic resonance (MR) imaging of the heart has, to date, been limited in its ability to evaluate cardiac function. The authors have implemented a technique for functional assessment of the heart using shorter echo times than those generally used for conventional spin-echo imaging. With these short echo times, multiple images can be obtained in a multisection mode approximately within the isovolumetric phases of the cardiac cycle. This permits a pair of image stacks to be obtained, one in end systole and the other in end diastole. With the use of a modified Simpson rule, left ventricular volume and ejection fraction were calculated and compared with results obtained from contrast material-enhanced ventriculography. Preliminary results indicate that this method has promise for the evaluation of a variety of functional parameters in the heart. The short acquisition times for this functional study permit it to be combined with a tissue characterization study within the time constraints of a clinical MR imaging session.

Heart Function Tests

Comparison of contrast x-ray biplane cineangiography and technetium-99m radionuclide scans for measurement of ventricular volumes in human autopsy hearts.

The area-length method is widely used in the determination of left ventricular volume. Although previous studies have reported that this technique overestimates true volume (TV), it is unknown whether this overestimation is the same at different volumes. In the present study, with the use of 10 postmortem human hearts, left ventricular volumes were determined by contrast x-ray biplane cine ventriculography (LVA), biplane radionuclide (LVR), and absolute-counts (LVC) technetium scans, and the results were correlated with TV. LVA correlated well with TV (r = 0.98). LVR correlated well with TV (r = 0.97), and LVR also correlated well with LVA (r = 0.96). Both area-length techniques (LVA and LVR) resulted in overestimation of TV with an upward shift of the regression line of 30.4ml +/- 3.8 (SEM) for LVA and 28.5 +/- 4.4 for LVR. The percentage of overestimation error was significantly greater at smaller left ventricular volumes (error = 33% at TV = 30ml and 10% at TV = 100ml, p less than 0.001). LVC correlated well with TV (r = 0.99), TVA, and LVR but underestimated TV. Thus left ventricular volumes can be reliably obtained from LVR and LVA by means of the area-length method and from LVC. When the area-length method is utilized, the percentage of error in the determination of left ventricular volume is proportional to 1/TV.

Angiocardiography

The syndrome of type A chronic atrophic gastritis, pernicious anemia, and multiple gastric carcinoids.

Gastric carcinoid tumors comprise approximately 1.0 to 2.7% of all carcinoid tumors. There appears to be an association between gastric carcinoids and pernicious anemia on the basis of underlying type A chronic atrophic gastritis. We review the relevant literature and describe a 63-year-old woman who had pernicious anemia and multiple polypoid gastric carcinoid tumors. The association of these disorders, proposed mechanisms of such association, and the need for an aggressive diagnostic approach are discussed.

Anemia, Pernicious

The discharge crisis: the experience of families of psychiatric patients.

Families of 16 psychiatric patients were interviewed immediately prior to discharge regarding the impending discharge of their relative, events prior to admission, and the families' impression of the hospitalization. Despite wide variation sample families' descriptors and differences in exposure to the treatment settings, there was a marked uniformity in responses. The families were not prepared for the discharge of their relative. In spite of their experiences and exposure to treatment, families continued to demonstrate tremendous uncertainty and lack of direction about the future with the patient. Families did not view the patient as ready to be discharged. They had not experienced an opportunity to express their views or participate in decision making. They did not consider the staff as a resource for themselves.

Aftercare