Bringing Molecular Biology to Bear on Adhesion Prevention: Postsurgical Adhesion Reduction Using Intraperitoneal Inoculation of Hyaluronic Acid–Inducing Adenoviral Vector in a Murine Model

Objective: Seprafilm (Genzyme, Cambridge, MA) an absorbable adhesion barrier incorporating hyaluronic acid (HA), a high molecular mass glycosaminoglycan and important component of the extracellular matrix, has been shown to prevent adhesions in both experimental models and human subjects. Yet, the application of HA as a sheet at the time of surgery has several important logistic limitations. Recently, our laboratory has identified and cloned the genes encoding murine hyaluronic acid synthase 2 (mHAS2) and 3 (mHAS3) and engineered adenoviruses incorporating these genes, which, on intraperitoneal injection, significantly increases HA in peritoneal fluid. We hypothesized that intraperitoneal gene therapy with mHAS2 or mHAS3 via an adenoviral vector prior to a standardized cecal abrasion surgery would lead to a reduction in postoperative adhesion severity. Methods: Mice were assigned to one of four groups: (1) intraperitoneal inoculation with adenovirus encoding mHAS2; (2) mHAS3; (3) a control reporter adenovirus (RV) encoding GFP; or (4) intraoperative placement of a commercially available and murine-validated hyaluronic acid adhesion barrier (Seprafilm, SF). An a priori sample size calculation was performed. Mice in groups 1, 2, and 3 underwent injection of 2 x 107 viral particles in 1 ml of fluid on day -1. Sham injection was performed on group 4 SF mice. On day 0, laparotomy was performed in random sequence by surgeon blinded to the experimental group. On day 7, adhesion scores (0-3) were assigned independently by two blinded investigators. Results: Mean adhesion scores (n = 247) were 0.68 (mHAS2), 0.91 (mHAS3), 1.28 (RV), and 0.47 (SF). Pairwise comparisons using Wilcoxon rank-sum test revealed significant reduction in severity of adhesions between mHAS2, mHAS3, and SF compared to RV (p = 0.0004, 0.039, and 0.0001, respectively). Significance persisted despite correction for multiple comparisons (p = 0.0002, Kruskal-Wallis). There was a direct relationship between intraperitoneal HA concentration and adhesion reduction. Only one death (RV) was secondary to adhesive disease; differential risk of death between groups was statistically significant (p = 0.008) (highest in mHAS2 group). Conclusions: In a dose-response relationship, an intraperitoneal gene therapy approach to adhesion prevention in a murine model was successful, with adenoviruses most productive of HA resulting in the most significant reduction in adhesion scores compared to empty virus (RV). Although SF best reduced postoperative adhesions, the adenoviral gene delivery approach may prove to be more effective in clinical use when peritoneal injury is less localized or at laparoscopy where the application of SF is not possible. Further studies to elucidate the reason for the differential death rates (time bias may have played a role) and to validate results are in progress

Impact of Zumba on Cognition and Quality of Life is Independent of APOE4 Carrier Status in Cognitively Unimpaired Older Women: A 6-Month Randomized Controlled Pilot Study

Objective: To investigate the association of a 6-month Zumba intervention with cognition and quality of life among older cognitively unimpaired apolipoprotein SMALL ELEMENT OF4 (APOE4) carrier and noncarrier women. Methods: Fifty-three women were randomly assigned to either twice-weekly Zumba group classes or maintenance of habitual exercise (control group) for 6 months. At baseline, 3, and 6 months, all participants underwent neuropsychological, physical activity, and quality-of-life assessments. Results: Overall, neuropsychological test scores and level of physical activity did not differ between intervention and control groups at any time. However, compared to the control group, quality of life was higher at 3 months, and visuospatial working memory and response inhibition improved more in the intervention group by 6 months. Apolipoprotein SMALL ELEMENT OF4 status did not affect the results. Discussion: Zumba may strengthen performance on visuospatial working memory among cognitively unimpaired older women but this needs to be tested in a larger clinical trial

Courtship, egg sac construction, and maternal care in Kukulcania hibernalis, with information on the courtship of Misionella mendensis (Araneae, Filistatidae)

Morphological and behavioural traits place Filistatidae basally within Araneomorphae, although some features, such as their continuing to moult after reaching adulthood, are reminiscent of mygalomorph spiders. This paper describes the courtship behaviour and other aspects of the reproductive biology of Kukulcania hibernalis and Misionella mendensis, and compares this information with that from related filistatid species and with Mygalomorphae. K. hibernalis has some unique behaviours during courtship (e.g. male lays threads on female web); other behaviours are probably widespread within Filistatidae (e.g. male uses the tarsi and metatarsi of one of his legs to rub the basal sections of the female’s legs and the sides of her cephalothorax). Some other behaviours seem more similar to Mygalomorphae than to those of other, more derived Araneomorphae. These include male construction of a large sperm web, and the positions of male and female facing each other during copulation, with the male holding the female cephalothorax lifted while insertions occur, similar to some mygalomorphs. The adult female K. hibernalis and the first instar spiderlings (outside the egg sac) feed simultaneously on the same prey, but spiderlings are also capable of cooperating during the attack of large prey. The courtship behaviour supports the hypothesis that places Filistatidae basally within Araneomorphae.UCR::Vicerrectoría de Docencia::Ciencias Básicas::Facultad de Ciencias::Escuela de Biologí

Reducing the cost of grammar-based testing using pattern coverage

In grammar-based testing, context-free grammars may be used to generate relevant test inputs for language processors, or meta programs, such as programming language compilers, refactoring tools, and implementations of software quality metrics. This technique can be used to test these meta programs, but the amount of sentences, and syntax trees thereof, which needs to be generated to obtain reasonable coverage of the input language is exponential. Pattern matching is a programming language feature used often when writing meta programs. Pattern matching helps because it automates the frequently occurring task of detecting shapes in, and extracting information from syntax trees. However, meta programs which contain many patterns are difficult to test using only randomly generated sentences from grammar rules. The reason is that statistically it is uncommon to directly generate sentences which accidentally match the patterns in the code. To solve this problem, in this paper we extract information from the patterns in the code of meta programs to guide the sentence generation process. We introduce a new coverage criterion, called Pattern Coverage, which focuses on providing a test strategy to reduce the amount of test necessary cases, while covering the relevant parts of the meta program. An initial experimental evaluation is presented and the result is compared with traditional grammar-based testing