HEF1 is a necessary and specific downstream effector of FAK that promotes the migration of glioblastoma cells.

The highly invasive behavior of glioblastoma cells contributes to the morbidity and mortality associated with these tumors. The integrin-mediated adhesion and migration of glioblastoma cells on brain matrix proteins is enhanced by stimulation with growth factors, including platelet-derived growth factor (PDGF). As focal adhesion kinase (FAK), a nonreceptor cytoplasmic tyrosine kinase, has been shown to promote cell migration in various other cell types, we analysed its role in glioblastoma cell migration. Forced overexpression of FAK in serum-starved glioblastoma cells plated on recombinant (rec)-osteopontin resulted in a twofold enhancement of basal migration and a ninefold enhancement of PDGF-BB-stimulated migration. Both expression of mutant FAK(397F) and the downregulation of FAK with small interfering (si) RNA inhibited basal and PDGF-stimulated migration. FAK overexpression and PDGF stimulation was found to increase the phosphorylation of the Crk-associated substrate (CAS) family member human enhancer of filamentation 1 (HEF1), but not p130CAS or Src-interacting protein (Sin)/Efs, although the levels of expression of these proteins was similar. Moreover downregulation of HEF1 with siRNA, but not p130CAS, inhibited basal and PDGF-stimulated migration. The phosphorylated HEF1 colocalized with vinculin and was associated almost exclusively with 0.1% Triton X-100 insoluble material, consistent with its signaling at focal adhesions. FAK overexpression promoted invasion through normal brain homogenate and siHEF1 inhibited this invasion. Results presented here suggest that HEF1 acts as a necessary and specific downstream effector of FAK in the invasive behavior of glioblastoma cells and may be an effective target for treatment of these tumors.

Frequent salad vegetable consumption is associated with a reduction in the risk of diabetes mellitus.

This cross-sectional study was undertaken to investigate the association between the reported frequency of consumption of vegetables and fruits, the choice of staple carbohydrate, and glucose intolerance. One thousand one hundred twenty-two subjects aged 40-64 years in a population-based study underwent an oral glucose tolerance test, and their food consumption was assessed using a food-frequency questionnaire. The crude prevalence of undiagnosed non-insulin-dependent diabetes mellitus (NIDDM) was 4.5%, and that of impaired glucose tolerance (IGT) 16.8%. The age-standardized prevalence rates were 2.3 and 11.2%, respectively. Frequent consumption of vegetables throughout the year was inversely associated with the risk of having NIDDM (odds ratio [OR] = 0.16; 95% confidence interval [CI] = 0.04-0.69). This association was maintained after adjustment for age, gender, and family history. Vegetable consumption during the summer months had a much weaker inverse association with the risk of having NIDDM that failed to reach statistical significance. A nonsignificant inverse association between frequent consumption of fruits and NIDDM was observed. Frequent self-reported pasta and rice consumption was associated with a reduction in the risk of having IGT and NIDDM. (OR = 0.62, 95% CI = 0.44-0.87, and OR = 0.51, 95% CI = 0.27-0.99, respectively) but this relationship was not independent of age. Whether these associations reflect specific effects of particular nutrients or are a reflection of the patterning of lifestyle factors remains to be determined.

Cigarette smoking is not associated with hyperinsulinemia: evidence against a causal relationship between smoking and insulin resistance.

Previous reports of a relationship between cigarette smoking and hyperinsulinemia and insulin resistance provide an important possible mechanism by which smoking could be associated with the metabolic cardiovascular syndrome and hence with ischemic heart disease. However, few previous studies have been able to adjust for all the possible confounding factors related both to smoking and to insulin resistance. Therefore, we examined this association in a population-based cohort study of 1,122 individuals aged 40 to 65 years who underwent a 75-g oral glucose tolerance test with specific measurement of insulin, 32,33-split proinsulin, and intact proinsulin concentrations. Physical activity was quantified using the Paffenbarger questionnaire, and smoking status and alcohol consumption were determined using the Health and Lifestyle Survey questionnaire; 17.4% of the population were current smokers and 32.4% were ex-smokers. Current smoking was associated with reduced overall obesity as indicated by the body mass index (BMI) but an increase in central adiposity as measured by the waist to hip ratio (WHR). There were also significant associations between cigarette smoking and the pattern of alcohol intake and physical inactivity. In unadjusted analyses, current smoking was associated with lower fasting and 120-minute insulin and also 120-minute glucose compared with levels in nonsmokers. Adjustment for confounding by age and BMI reduced these differences, but they were increased by adjustment for central obesity. We conclude from this study that a causal relationship between cigarette smoking and insulin resistance is unlikely.

Carbon monoxide levels in the breath of smokers and nonsmokers: effect of domestic heating systems.

Breath and ambient (room) carbon monoxide (CO) levels were measured in a random sample of 168 adults in their own homes. The levels of breath CO in the 69 smokers ranged from 3 ppm to over 100 ppm, 74% being above 10 ppm; mean levels in the 99 nonsmokers were lower than in the smokers, 79% being below 6 ppm. In the remaining 21% of nonsmokers with higher breath levels than expected, the ambient CO was also found to be elevated, ranging up to 38 ppm. A close correlation in the nonsmokers was found between the breath and ambient CO levels (r = 0.952, p less than 0.001). The rooms with the elevated ambient CO levels (above 5 ppm) were those which, at the time of testing, were being heated by gas radiant heaters, open fires or stoves. The maximum ambient CO in the rooms of smokers with non CO generating heating was 16 ppm. The results suggest that many people, both smokers and nonsmokers, may be at risk from CO generated by certain domestic heating systems and that nonsmokers are far more likely to be exposed to high levels of CO from these sources than from being in a room with a heavy smoker. Poor ventilation associated with the current trend towards excluding all draughts is likely to exacerbate the situation for both smokers and nonsmokers.

Some regional variations in dietary patterns in a random sample of British adults.

Comparison was made of the reported frequency of consumption or choice of 30 food items by 8860 adults in the 11 standard regions of Great Britain, with the use of log-linear analysis to allow for the age, sex, social class and smoking habit variations between the regions. The South-East was taken as the base region against which the others were compared. The number of food items for which there were significant differences from the South-East were Scotland 23, North 25, North-West and Yorkshire/Humberside 20, Wales 19, West Midlands 15, East Midlands 10, East Anglia 8, South-West 7 and Greater London 9. Overall the findings confirm a North/South trend in relation to eating habits, even when demographic and smoking-habit variations are taken into account, with the frequent consumption of many fruit and vegetable products being much less common and of several high-fat foods (chips, processed meats and fried food) more common in Scotland, Wales and the northern part of England. In most regions there was a significantly lower frequency of consumption of fresh fruit, fruit juice, 'brown' bread, pasta/rice, poultry, skimmed/semi-skimmed milk, light desserts and nuts, and a higher consumption of red meat, fish and fried food than in the South-East.

The induction by X-rays of chromosome aberrations in male Guinea-pigs, rabbits and golden hamsters. III. Dose-response relationship after single doses of X-rays to spermatogonia.

The induction by X-rays of translocations in spermatogonia was studied by cytological means in spermatocytes derived from them. In the rabbit and guinea-pig hump shaped dose-response curves were obtained, with a linear relationship at the low doses. The shapes of the curves were similar to those reported for the mouse, except that the maximum occurred at 600-700 rad in the mouse as opposed to 300 rad in the guinea-pig and rabbit. Unlike the guinea-pig and rabbit, the golden hamster showed a hump dose-response curve without a definite peak value and with little decrease in yield at high radiation doses. Over the low dose range 100-300 rad, the slopes of the curves of translocation yield were in the order:mouse (highest), rabbit, guinea-pig and hamster. Data on sterile periods suggested that the amount of spermatogonial killing in the rabbit and guinea-pig was as great or greater than in the mouse, and that in the golden hamster it was most severe. It is suggested that the differing shapes of the dose-response curves can be explained by a lower sensitivity to translocation induction in the test species and, also especially in the golden hamster, a greater sensitivity to cell killing. The possibility of extrapolating from these data to other species is discussed.

The induction by X-rays of chromosome aberrations in male Guinea-pigs and golden hamsters. IV. Dose response for spermatogonia treated with fractionated doses.

The effect of dose fractionation on the induction of translocations by 400 and 600 rad X-rays in spermatogonia of guinea-pigs and hamsters was investigated cytologically. Three types of fractionation were used, dividing the dose into (a) two equal fractions 24 h apart, (b) two equal fractions 8 weeks apart, and (c) eight or twelve equal fractions of 50 rad, at intervals of one week. The two species responded similarly throughout, but gave lower translocation yields than the mouse. The effects of the first and third types of fractionation were similar to those described previously in the mouse, and suggested that a first radiation dose modifies the spermatogonial population so that its sensitivity to a dose 24 h later is altered, and that repeated radiation doses result in development of resistance to translocation induction. After 8-week fractionation the results suggested that in guinea-pigs and hamsters the spermatogonial population had not returned to normal by 8 weeks after the first dose, whereas in the mouse normal sensitivity had returned by this time. The results, reported previously, of single doses of X-rays suggest that the spermatogonial population consists of fractionated doses in the mouse suggest that the sensitive and resistant types represent different phases of the same cell type rather than two distinct types of cell. In the guinea-pig and hamster this question remains open.

X-ray induced dominant lethal mutations in mature and immature oocytes of guinea-pigs and golden hamsters.

The induction of dominant lethal mutations by doses of 100-400 rad X-rays in oocytes of the guinea-pig and golden hamster was studied using criteria of embryonic mortality. For both species higher yields were obtained from mature than from immature oocytes, in contrast to results for the mouse. Data on fertility indicated that in the golden hamster, as in the mouse, immature oocytes were more sensitive to killing by X-rays than mature oocytes but that the converse was true in the guinea-pig. The dose-response relationship for mutation to dominant lethals in pre-ovulatory oocytes of guinea-pig and golden hamsters was linear, both when based on pre- and post-implantation loss and when on post-implantation loss only. The rate per unit dose was higher for the golden hamster, and the old golden hamsters were possibly slightly more sensitive than young ones. The mutation rate data for mature oocytes of the mouse, using post-implantation loss alone, also fitted a linear dose-response relationship, except that the rate per unit dose was lower than for the other two species.

The induction by X-rays of chromosome aberrations in male guinea-pigs, golden hamsters and rabbits. II. Properties of translocations induced in post-meiotic stages.

Translocations induced by X-rays in post-meiotic germ cells of male guinea-pigs, golden hamsters and rabbits were studied cytologically in the F1 sons of the irradiated males. The percentage of spermatocytes displaying multivalent configurations varied with the translocation, but the average percentage appeared to depend on the species: fewer quadrivalents were observed in hamster than in guinea-pig heterozygotes and most were recorded for rabbit heterozygotes. Chain quadrivalents were more abundant than ring quadrivalents at meiosis for the guinea-pig and hamster, in contrast to the mouse. Too few translocation heterozygotes were examined to determine which meiotic configuration was the more prevalent in the rabbit. In all three species, as in the mouse, translocations were found which caused male sterility, due to partial or complete failure of spermatogenesis, although most translocations caused semi-sterility. For these semi-sterile males both the frequency and time of embryonic death in the progeny appeared to be the same as in the mouse. It is concluded that similar types of chromosome aberrations are induced by X-rays in post-meiotic germ cells of male guinea-pigs, rabbits, golden hamsters and mice.

The induction by X-rays of chromosome aberrations in germ cells of male guinea-pigs, golden hamsters and rabbits. I. Dose response in post-meiotic stages.

The induction by X-rays of translocations in post-meiotic germ cells of the guinea-pig, golden hamster and rabbit was studied by cytological analysis of male offspring of the irradiated animals. As reported previously for the mouse, the pattern of sensitivity to dominant lethal induction, as indicated by litter-size, was similar to that for translocation induction in both the guinea-pig and golden hamster. In both speciesspermatids were more sensitive than spermatozoa, and in the golden hamster spermatocytes gave a lower yield than spermatids. The translocation frequency among post-meiotic germ cells treated with 600 rad was higher in the rabbit than the guinea-pig, and both were above that for the golden hamster. However, for spermatozoa, species differences with respect to the recovered translocation yield appeared to depend on dose. In the hamster, the translocation frequency after 600 rad, as measured in the female offspring, was similar to that obtained in the male offspring. A small amount of data on the induction of sex-chromosome aneuploidy by 200 rad in golden hamsters suggested that the hamster might be as sensitive as the mouse.

Dose-response data for X-ray induced translocations in spermatogonia of Rhesus monkeys.

The yields of translocations in spermatocytes after irradiation of spermatogonia of Rhesus monkeys with doses of 100, 200 or 300 rad X-rays were low, and consistent with a humped dose-response curve with a peak at about 200 rad. Such a curve would agree well with earlier results on the marmoset and man, but the yields at any dose in the Rhesus monkey were lower.

Associations between off-label feed additives and farm size, veterinary consultant use, and animal age.

Data from the United States National Swine Survey collected by the National Animal Health Monitoring System were used to describe the use of feed additives in swine feeds. Data were collected from 710 farms. The concentration of feed additives expressed in grams per ton of complete feed was described by stage of production, and the use of feed additives above the labeled treatment levels (i.e. off-label) was identified. Of the 3328 feeds, about 79% contained feed additives used in the labeled manner. For all classes of pigs, the prevalence of labeled feed additive use was greater than 75%. Penicillin was used according to its label most often, followed by apramycin, bacitracin, tetracyclines, lincomycin, and tylosin. Carbadox had the highest prevalence of off-label use. Of the 699 feeds that included feed additives in an off-label manner, about 57% included additives at greater than the recommended concentrations or were fed to an incorrect class of pig. About 56% of the feeds had off-label combinations of additives. Small farms were more likely to use rations with no feed additives than intermediate or large farms (P < 0.001). Of those farms using feed additives, the odds of a small farm using all feed additives in the labeled manner was 7.7 times that of an intermediate or large farm (P < 0.0001). After controlling for herd size, producers who used a veterinary consultant were 2.1 times more likely to use feeds with feed additives (P < 0.0001).

Effect of ankle orthoses on functional performance for individuals with recurrent lateral ankle sprains.

Ankle orthoses are commonly used for protection against initial ankle sprain injuries and for protection against reinjury. The purpose of this study was to compare the effects of the DonJoy Ankle Ligament Protector (ALP) and the Aircast Sport-Stirrup (AS) on the functional performance tasks of a 40-meter sprint, a figure-of-eight run, and standing vertical jump. Subjects were 14 males and nine females who reported a history of two or more lateral ankle sprains of the same ankle and no sprains of the contralateral ankle. Each subject performed all functional tasks on the first test day with both ankles unbraced and with the recurrently sprained ankle braced with one of the two orthoses. Each subject returned for testing on a second day and performed all tasks with both ankles unbraced and with the other orthosis on the recurrently sprained ankle. Subjects also rated the orthoses for comfort and support. Analysis of variance indicated that braced data were not significantly different than unbraced data and that data for the two orthoses did not differ significantly for any of the functional performance tasks. Seventy-four percent of the subjects reported the AS was more comfortable than the ALP, and 61% of the subjects indicated that the AS provided more support than the ALP. Neither orthosis had an effect on functional performance. The results may assist clinicians in selecting either of these orthoses for use in protection against ankle sprain injury.

Children's use of mnemonic strategies: variability in response to metamemory training.

In this study, 8- to 12-year-old children were tested to determine their level of use of a strategy for organizing lists of low-associated words into semantic groups. Then, on the basis of their scores in the strategy tests, the children were placed in either a strategic or nonstrategic group. The groups were divided into a sort-instruction training condition, in which they were told to group words so that they "go together" in some way, and a metamemory training condition, in which these instructions were supplemented by feedback, strategy reminders, and opportunities to self-test. One week later, each child's strategy use was reassessed. Children in the strategic group sorted semantically throughout training and transfer, and they showed high recall, regardless of condition. Children in the nonstrategic group showed high strategy use and recall during training, but they showed significantly more variability in recall than did the children in the strategic group. Children in the nonstrategic-metamemory condition who benefited from training were also more likely to transfer the use of the strategy than were children trained in the sort-instruction condition.

Effects of temporally varied biased and unbiased story summaries on the suggestibility of preschoolers.

Preschool children's resistance to the influence of postevent misinformation was investigated. Ninety-one preschoolers (3 years 6 months to 5 years 6 months old) were read a story about a boy's birthday party. One week later, they received 1 of 3 summary conditions containing general information (unbiased, biased, or no summary). Two weeks after the original story, they were presented with either a biased summary or no summary. The children's suggestibility was assessed by a recognition test that provided a choice between the original and postevent misinformation. Those who received an unbiased summary showed higher recognition rates than those who received a biased summary, regardless of whether the biased information had been given at the 1- or 2-week interval. Although an unbiased summary followed by biased information did not produce greater recognition rates that were found in the control group, participants who received unbiased summaries did perform above chance, suggesting that even a vague summary may help to reactivate specific memory traces, particularly when information is bimodally presented.