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Biomedical subjects

Adam Harris

Publications and source records attributed to Adam Harris.

5 recordsLinked to original sources

Rearrangement of a conditional allele regardless of inheritance of a Cre recombinase transgene.

To generate conditional gene knockouts in osteoblasts, we previously developed transgenic mice in which Cre recombinase cDNA was cloned downstream of a 3.6 or 2.3 kb fragment of the rat Col1a1 promoter (Col3.6-Cre and Col2.3-Cre, respectively). Col-Cre mice were bred with mice in which exon 4 of the Igf1 gene is flanked by loxP sites. Mating units were arranged such that either the male or the female breeder transmitted the Col-Cre transgenes. Progeny were evaluated for Cre-mediated Igf1 gene rearrangement. We found that the loxP-flanked Igf1 locus was rearranged in the absence of inheritance of the Cre transgene. The incidence was 50 and 28% with Col2.3-Cre and Col3.6-Cre females, respectively, and 15 and 18% with Col2.3-Cre and Col3.6-Cre males, respectively.

Alleles↗

Mortality after epilepsy surgery.

Mortality rates are higher in people with refractory epilepsy than in the general population. We assessed mortality rates in a prospectively followed cohort who had epilepsy surgery, to examine the factors related to mortality and to assess the relationship between seizure control and mortality. Five hundred eighty-three patients were evaluated. Mortality was strongly related to seizure control (p = 0.001), with 18 deaths observed in patients with recurrent seizures (mortality rate = 11.4 per 1,000 person-years) and 1 death in patients with no recurrent seizures (mortality rate = 0.85 deaths per 1,000 person-years). Patients with generalized epilepsy who had corpus callosotomy had a higher mortality rate than patients who had resective or transective surgery. The side of surgery and gender did not influence mortality rates. The standardized mortality ratio was 5.75 for patients with recurrent seizures and was significantly higher for females than males. These data show that the excess mortality associated with refractory epilepsy is eliminated after epilepsy surgery when seizures are abolished and suggest that epilepsy surgery reduces the risk of epilepsy-associated death.

Cohort Studies↗

Advanced transfection with Lipofectamine 2000 reagent: primary neurons, siRNA, and high-throughput applications.

Lipofectamine 2000 is a cationic liposome based reagent that provides high transfection efficiency and high levels of transgene expression in a range of mammalian cell types in vitro using a simple protocol. Optimum transfection efficiency and subsequent cell viability depend on a number of experimental variables such as cell density, liposome and DNA concentrations, liposome-DNA complexing time, and the presence or absence of media components such as antibiotics and serum. The importance of these factors in Lipofectamine 2000 mediated transfection will be discussed together with some specific applications: transfection of primary neurons, high throughput transfection, and delivery of small interfering RNAs.

Animals↗