Insertion of a nuclear factor kappa B DNA nuclear-targeting sequence potentiates suicide gene therapy efficacy in lung cancer cell lines.

Lung cancer currently causes the majority of cancer-related deaths worldwide and new treatments are in high demand. Gene therapy could be a promising treatment but currently lacks sufficient efficiency for clinical use, primarily due to limited cellular and nuclear DNA delivery. In the present study, we investigated whether it was possible to exploit the endogenous nuclear-shuttling activity by the nuclear factor kappa B (NFκB) system, which is highly prominent in many cancers as well as lung cancer. We observed that insertion of a DNA nuclear-targeting sequence (DTS) recognized by NFκB could improve plasmid nuclear delivery and enhance the therapeutic effect of a validated transcriptionally cancer-targeted suicide gene therapy system. A clear correlation between the number of inserted NFκB-binding sites and the therapeutic effect of the suicide system was observed in both small cell lung cancer (SCLC) and non-SCLC cell lines. The effect was observed to be due to elevated nuclear translocation of the suicide gene-encoding plasmids. The results show that a significant improvement of gene therapeutic efficiency can be obtained by increasing the intracellular trafficking of therapeutic DNA. This is to our knowledge the first time a DTS strategy has been implemented for suicide gene therapy.

Viral vector-mediated gene expression in olfactory ensheathing glia implants in the lesioned rat spinal cord.

Implantation of olfactory ensheathing glia (OEG) is a promising strategy to augment long-distance regeneration in the injured spinal cord. In this study, implantation of OEG following unilateral hemisection of the dorsal cervical spinal cord was combined with ex vivo gene transfer techniques. We report, to our knowledge for the first time, that purified cultures of primary OEG are capable of expressing a foreign gene following adenoviral (AdV) and lentiviral (LV) vector-mediated gene transfer. OEG implants subjected to AdV vector-mediated gene transfer expressed high levels of transgenic protein in both intact and lesioned spinal cord at 7 days after implantation. However, the levels of transgene expression gradually declined between 7 and 30 days after implantation in lesioned spinal cord. Infection with LV vectors resulted in stable transduction of primary OEG cultures and transgene expression persisted for at least 4 months after implantation. Genetic engineering of OEG opens the possibility of expressing additional neurotrophic genes and create optimal 'bridging' substrates to support spinal axon regeneration. Furthermore, stable transduction of OEG allows us to reliably study the behaviour of implanted cells and to obtain better understanding of their regeneration supporting properties.

[Complications in two children with acute lymphatic leukemia caused by vaccination against varicella zoster virus]. / Komplikationer hos to børn med akut lymfatisk leukaemi vaccineret mod Varicella zoster-virus.

Complications in two varicella zoster virus (VZV) vaccinated children with leukemia in remission are reported. Case I presented with varicella on day 30 after vaccination, with a relapse on day 49 and development of zoster on day 70. VZV was detected in vesicles by PCR on days 49 and 70. Case II presented with varicella on day 32 after vaccination, and VZV was detected in vesicles and nasal secretion. The manifestations were mild and responded to treatment. PCR methods were directed toward the R5 and PS regions. The virus from the two children was unambiguously identified as the Oka vaccine strain. The majority of Danish field strains had only one copy of the 112 basepair repeat element in the R5 region, but two, four and presumably higher copy numbers were also seen. All Danish field strains had the Pst1 cleavage site in the PS region.

Individualized gonadotropin regimens for follicular stimulation in macaques during in vitro fertilization (IVF) cycles.

Follicular stimulation was compared in macaques receiving sequential gonadotropin treatment which was terminated after seven, eight, or nine days depending on the time required to attain preselected criteria of follicular maturation. Although estradiol levels and follicle sizes varied, the number of follicles and oocytes/animal, oocyte nuclear maturity, IVF rates and progesterone levels during the luteal phase were similar among groups. Reducing the duration of gonadotropin treatment to individualize follicular stimulation regimens does not compromise follicle or gamete quality.

The physiological effects of step training with and without handweights.

The purpose of this study was to examine the physiological effects of eight weeks of step training with (N = 12) and without (N = 12) handweights. Twenty-four college-aged females volunteered as subjects for the study which involved pre- and post-laboratory testing consisting of the following: an incremental treadmill test for determination of aerobic work indices, which included maximal oxygen uptake rate (VO2max), maximal heart rate, ventilatory threshold oxygen uptake rate (VT-VO2), and ventilatory threshold heart rate (VT-HR); hydrostatic weighing for determination of body composition, which included percent body fat (%BF), fat mass (FM), and lean body mass (LBM); anthropometric measurements for determination of body build characteristics, which included the endomorphic (ENDO) somatotype rating, mesomorphic (MESO) somatotype rating, and ectomorphic somatotype rating; isokinetic strength testing at 60 degrees.sec-1 for determination of flexion and extension strength of the arm at the shoulder joint, forearm at the elbow joint, and leg at the knee joint; and the concentric leg press for determination of overall lower body strength. Two-way analysis of covariance indicated that the main effect of step training resulted in significant (p < 0.05) overall improvements in VO2max (38.29 +/- 1.05 to 41.32 +/- 0.95 ml.kg-1/min-1), VT-VO2 (22.86 +/- 0.86 to 26.03 +/- 0.84 (ml.kg-1/min-1), VT-HR (159.62 +/- 3.01 to 168.29 +/- 3.19 b/min-1), %BF (26.16 +/- 1.05 to 23.95 +/- 1.06%), FM (15.13 +/- 0.96 to 13.82 +/- 0.96 kg), ENDO rating (5.24 +/- 0.18 to 4.93 +/- 0.18 units), MESO rating (3.22 +/- 0.19 to 3.50 +/- 0.19 units), arm flexion strength (30.73 +/- 1.83 to 35.08 +/- 1.73 N/m), forearm flexion strength (26.89 +/- 1.13 to 29.21 +/- 1.14 N/m), and forearm extension strength (28.13 +/- 1.26 to 31.07 +/- 1.38 N/m). Similar improvements were observed in both the handweight and no-handweight groups of subjects.

Physical characteristics of novice and experienced women marathon runners.

The physiological and anthropometric characteristics of 23 non-elite women marathoners were studied. Ten of these women had never run a marathon before (novices) and 13 had run at least one marathon during the previous year (experienced). A comparison of characteristics of these two groups to each other and to elite women marathoners, as reported in the literature, disclosed no significant (p greater than 0.01) among the groups in age, % body fat, body weight, height, lean body mass or HR max. Significant differences (p less than 0.01) were noted, however, in VO2 max (45.8, 51.8, 59.1 ml.kg-1 min-1), VE max (76.3, 94.7, 108.9 L.min-1), and years of training (0.54, 2.06, 4.55 years) with the novice runners having the smallest values, the experienced runners having the next larger values, and the elite runners having the largest values. For our subjects, estimated percentage of body fat did not correlate with finish time, but VO2 max and finish time were significantly related (r = -0.72, p less than 0.01). This suggest that women marathon runners are similar in anthropometric measurements, and that improved performance is associated with higher aerobic capacity and years of training rather than with body dimensions.

Effects of three kinds of hypoxias on vigilance performance.

The consequences for vigilance performance of inhaling carbon monoxide at altitude were examined. In addition, the effects of altitude and CO at an equivalent reduction in O2-carrying capacity were compared. Ten subjects performed a visual vigilance task under four atmospheric conditions. Physiological measures included heart rate, blood pressure, and ventilation. Blood hemoglobin and CO levels were measured directly from blood samples drawn after 0,50 and 120 min of exposure. The physiological variables measured and subjective responses of subjects showed no changes attributable to the atmospheric conditions. A statistically significant change in vigilance performance as measured by precentage of signals detected, was found between control and low oxygen; however, performance under CO and the combination of CO and low oxygen was not different from control. The lack of deterioration in performance under the combination of CO and low oxygen suggested that the increased severity of the stress resulted in activation of compensatory mechanisms which counterbalanced the decreased O2 available to the tissues.