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Biomedical subjects

R M el Oakley

Publications and source records attributed to R M el Oakley.

11 recordsLinked to original sources
PubMed1998

Efficiency of a high-titer retroviral vector for gene transfer into skeletal myoblasts.

BACKGROUND: Genetic transformation of skeletal myoblasts for myocardial repair is dependent on an efficient gene transfer system that integrates the genes of interest into the genome of the target cell and its progeny. The aim of this investigation was to evaluate the use of a new retrovirally based gene transfer system for this purpose. METHODS: MFGnlslacZ retroviral vector, packaged in high-titer, split-genome packaging cell line (FLYA4) was used to transduce the skeletal myoblast cell line L6. L6 cells, cultured in 10% fetal calf serum, were transduced with the MFGnlslacZ vector by means of filtered supernatant from FLYA4 cells. Transduced L6 cells were divided into four groups. Group I cells were fixed as myoblasts 3 days after transduction. Group II cells were allowed to differentiate into myotubes. Group III cells were split every 3 days for 4 months. Group IV cells were split as in group III but then allowed to differentiate into myotubes. All samples were fixed and stained for beta-galactosidase activity. The effects on gene transfer of transforming growth factor-beta, insulin-like growth factor-I, and platelet-derived growth factor were determined by spectrophotometric assay of beta-galactosidase activity in cells transduced in the presence or absence of serum with 0 to 200 ng/ml of each growth factor. RESULTS: Morphometric analysis showed that 66.3% +/- 3% to 69.6% +/- 6% of cells in group I to IV expressed the lacZ reporter gene. In the presence of serum, transforming growth factor-beta significantly inhibited gene transfer, whereas insulin-like growth factor-I and platelet-derived growth factor significantly enhanced gene transfer. In absence of serum, however, only platelet-derived growth factor enhanced retrovirally mediated gene transfer into skeletal myoblasts. CONCLUSION: MFG retroviral vectors packaged in FLYA4 cells are efficient in gene transfer into skeletal myoblasts and result in transgenic expression that is maintained after repeated cell division, differentiation, or both. Platelet-derived growth factor enhances retrovirally mediated gene transfer into skeletal myoblasts.

Cell Differentiation↗
PubMed1996

Extended criteria for cardiac allograft donors: a consensus study.

BACKGROUND AND METHODS: To achieve a consensus regarding extending the current criteria for referring potential cardiac allograft donor, we sent a detailed questionnaire to all heart transplant surgeons in the United Kingdom. RESULTS: Replies representing the opinion of 16 heart transplant surgeons (88% response rate) suggest that the majority believe that donor age could be extended up to 55 years even in the absence of a coronary angiogram or echocardiogram. Family history of ischemic heart disease, history of hyperlipidemia or smoking, and brain death caused by paracetamole or barbiturates overdose were not considered prohibitive to cardiac allograft donation. Similarly, chest trauma, prolonged mechanical ventilation, pre-referral cardiac arrest or hypotension for any length of time, and nonspecific ST-segment elevation were not considered to be contraindications. By contrast, significant Q waves in a recent electrocardiogram, the presence of hepatitis C antibodies, pulmonary capillary wedge pressure more than 20 mm Hg, and dependency on multiple inotropic support were considered definite exclusion criteria. Donor hormonal resuscitation (T3-cocktail), insertion of a Swan-Ganz catheter, and direct measurement of intracavitary pressure at the time of retrieval were considered unnecessary by more than 80% of respondents. There was no consensus of opinion regarding the use of donors above the age of 55 years, brain death caused by carbon monoxide poisoning, history of diabetes mellitus, alcohol or drug abuse, systemic infections, electrocardiographic evidence of left ventricular hypertrophy, or the maximum dose of inotropic support required to maintain satisfactory hemodynamics. CONCLUSIONS: The number of potential cardiac allograft donors can be increased by extending the donor age to 55 years. Brain death caused by Paracetamole and barbiturate overdose may not prohibit organ donation.

Adolescent↗
PubMedDate not supplied

Effects of cardiomyoplasty on cardiac growth in rats.

BACKGROUND AND AIM OF THE STUDY: Cardiomyoplasty (CMP) has been proposed as a treatment for pediatric patients, but restriction of cardiac growth by the muscle wrap is a potential source of concern. This possibility was investigated in an immature animal model. METHODS: Six-week-old rats (body weight 203.8 +/- 5.4 g, mean +/- SEM) underwent either left thoracotomy with CMP (group I, n = 7), or thoracotomy without CMP (group II, n = 8). A third group (group III, n = 7) served as untreated controls. Final measurements were made 20 weeks later after body weights had reached a plateau. RESULTS: Preoperative body weights were not significantly different between the groups. At elective sacrifice, the body weights of animals that underwent surgery did not differ significantly (group I, 558.0 +/- 21.5 g and group II, 617.3 +/- 20.3 g), but were significantly less than those of control animals (727.6 +/- 13.3 g, p < 0.001 and p < 0.01, respectively). Cardiac ventricular weights in the CMP group were significantly less than those of control animals (group I, 1.21 +/- 0.06 g; group III 1.45 +/- 0.04 g; p < 0.01), but were not statistically different from those of the sham thoracotomy group (group II, 1.36 +/- 0.05 g). Mean left ventricular end-diastolic volumes were similar in all groups (group I, 0.67 +/- 0.07 mL; group II, 0.66 +/- 0.07 mL; and group III, 0.69 +/- 0.10 mL; p = ns). CONCLUSIONS: A major surgical procedure impairs growth in juvenile rats. no evidence emerged from this study for additional restriction of cardiac development due to cardiac wrapping. However, studies that include stimulated muscle wraps are needed before CMP should be considered for the pediatric age group.

Animals↗
PubMedDate not supplied

Factors affecting the integrity of latissimus dorsi muscle grafts: implications for cardiac assistance from skeletal muscle.

BACKGROUND: Severe latissimus dorsi muscle damage may compromise cardiomyoplasty performance. We analyzed factors underlying the damage produced in 20 sheep latissimus dorsi muscles by isolating the influences of electrical stimulation, mobilization (with some loss of vascular supply), loss of normal resting tension, or a combination of these. METHODS: In group I (n = 3), the muscle was mobilized except for its neurovascular pedicle and reattached at normal resting length. In group II (n = 3), the muscle was mobilized and reattached at about 80% of resting length. Groups III (n = 6) and IV (n = 4) were as groups I and II except that continuous indirect stimulation at 2 Hz was added after 2 weeks. In group V (n = 4), the undisturbed muscle received stimulation alone. After 10 to 12 weeks, muscle samples were taken for morphometric analysis. RESULTS: Loss of resting muscle tension appeared to be the single most damaging intervention, though mobilization and stimulation had further deleterious effects. The worst damage was seen when all three factors were combined, when 60% of the muscle cross section was occupied by connective tissue and fat. The changes were significantly more severe in the distal than in the proximal part of the muscle, implicating ischemia as a contributory factor. CONCLUSIONS: Fiber damage reduces the effectiveness of muscle grafts used for cardiac assistance and merits further systematic investigation.

Adipose Tissue↗