Magnetic perturbation diagnostics in the high-temperature lithiated environment of LTX-ß.

Magnetic perturbation measurements will be invaluable for characterizing Lithium Tokamak Experiment Beta (LTX-ß) plasmas due to the time-evolving 3D nature of the magnetic fields generated by eddy currents in the vessel and copper shell segments, as well as enhanced MHD instability drive due to newly introduced neutral beam heating. The LTX-ß upgrade includes two new arrays of Mirnov coils: a shell eddy sensor array of two-axis coils distributed over the back surface of one shell segment and a toroidal array of poloidal field coils at the low-field side midplane gap. Evaporative lithium wall-coating and the high temperatures required for liquid lithium wall operation both complicate the implementation of in-vessel diagnostics. While the shell array is protected from lithium exposure, the shell segment to which it is mounted will at times exceed 300 °C. The toroidal array, however, will experience direct line-of-sight exposure to the lithium evaporator as well as close proximity to the hot shell and may also be subject to poorly confined beam-driven fast ions. We describe how the two new Mirnov coil arrays meet these environmental challenges and enhance the LTX-ß diagnostic suite.

Dietary ractopamine supplementation during the first lactation affects milk composition, piglet growth and sow reproductive performance.

Excessive mobilization of body reserves during lactation delays the return to reproductive function in weaned primiparous sows. This study tested the hypothesis that supplementing the lactation diets of first-parity sows with ractopamine hydrochloride would reduce maternal weight loss and improve subsequent reproductive performance. Gestating gilts were allocated to one of two treatment groups (n=30 sows/treatment), with one group fed a standard lactation diet (2.5g/Mcal LYS: DE) throughout lactation (CTRL), whereas the treatment group received the standard lactation diet supplemented with 10mg/kg ractopamine hydrochloride (RAC) from d 1 to 13 of lactation and 20mg/kg RAC from d 14 of lactation until artificial insemination (AI). Weaning occurred on d 21 of lactation, with AI occurring at the first post-weaning estrus. Compared to CTRL, RAC supplementation decreased (P<0.05) liveweight loss between d 13 and 20 of lactation (4.3±0.90 versus 1.3±0.96kg), and tended to increase (P=0.06) the number of second litter piglets born alive (9.5±0.52 versus 8.1±0.74). Treatment (RAC versus CTRL) reduced milk protein levels on d 13 and 20 of lactation (P<0.05), and piglet weight gain between d 13 and 20 of lactation (260±0.01 versus 310±0.01g/day, P<0.01). In conclusion, it is evident that dietary RAC altered milk composition and stimulated conservation of maternal body reserves during the third week of lactation, resulting in a beneficial effect on subsequent reproductive performance.

Group and individual sow behavior is altered in early gestation by space allowance in the days immediately following grouping.

Aggression between domestic sows is greatest when sows are first introduced to each other and hierarchies form. The aim of this study was to determine the effect of a spacious "mixing pen" on sow aggression and stress. Sows were mixed into groups of 6 and allowed 2 (LOW; 8 groups and 48 sows), 4 (MED; 7 groups and 42 sows), or 6 m/sow (HIGH; 7 groups and 42 sows) for 4 d after mixing, at which point all pens were equalized to 2 m/sow. Salivary cortisol concentration and injury counts were measured on d -1, 0, 1, 3, and 4 relative to mixing, and behavior was also recorded on each of these days following mixing. Reproductive performance was assessed at farrowing. A linear mixed model was applied to the data. Data are presented as least squares means and standard error of the mean. Where transformations occurred, nontransformed adjusted means are presented in parentheses following the presentation of transformed data. In the primary analyses where measures were considered at the pen level, there were no effect of space allowance on fight number per sow, duration of fights, percentage of total time spent fighting, displacements, bites, knocks, and lunges ( > 0.05). These measures were higher on d 0 (i.e., fight number 1.0 ± 0.1 [13.8]) compared with d 1 (0.4 ± 0.1 [4.2]), 3 (0.7 ± 0.1 [5.3]), and 4 (0.7 ± 0.1 [5.5]; < 0.05), with no increase in aggression on d 4 when pen sizes were standardized ( > 0.05). There was increased percentage of time spent active (1.5 ± 0.02 [33.7] for LOW, 1.5 ± 0.02 [36.5] for MED, and 1.6 ± 0.02 [43.4] for HIGH) and time spent exploring (1.8 ± 0.1 [3.5] for LOW, 2.0 ± 0.1 [4.0] for MED, and 2.3 ± 0.1 [5.7] for HIGH) and number of nonaggressive sow-sow contacts (0.3 ± 0.09 [2.2] for LOW, 0.4 ± 0.07 [3.2] for MED, and 0.5 ± 0.07 [4.5] for HIGH) in HIGH compared with LOW ( < 0.05). Farrowing rate and total piglets born were not affected by treatment ( > 0.05). A secondary analysis was conducted that examined individual sow behavior within each pen, and this identified increased injury number in the lowest ranked sows (involved in no fights on d 0 and no displacements on d0 to d4) in LOW (9.3 ± 1.2 [107.9] for LOW, 6.2 ± 0.8 [53.0] for MED, and 5.1 ± 0.8 [28.1] for HIGH) and also decreased fight number and duration in HIGH compared with LOW on d 0 and 1 ( < 0.05). Our primary data analysis demonstrates positive exploratory and social behaviors with increased space and suggests that a reduction in space following hierarchy formation is not a significant stressor. Additionally, there is some evidence at an individual sow level that increased space at mixing benefits sow welfare parameters, especially for low-ranked sows.

Design and installation of a ferromagnetic wall in tokamak geometry.

Low-activation ferritic steels are leading material candidates for use in next-generation fusion development experiments such as a prospective component test facility and DEMO power reactor. Understanding the interaction of plasmas with a ferromagnetic wall will provide crucial physics for these facilities. In order to study ferromagnetic effects in toroidal geometry, a ferritic wall upgrade was designed and installed in the High Beta Tokamak-Extended Pulse (HBT-EP). Several material options were investigated based on conductivity, magnetic permeability, vacuum compatibility, and other criteria, and the material of choice (high-cobalt steel) is characterized. Installation was accomplished quickly, with minimal impact on existing diagnostics and overall machine performance, and initial results demonstrate the effects of the ferritic wall on plasma stability.

Melatonin implants do not alter estrogen feedback or advance puberty in gilts.

Puberty in pigs is often delayed during late summer and autumn, with long daylength the most likely cause. We hypothesised (1) that gilts born around the shortest day would have a later release from the negative feedback actions of estradiol than gilts born around the spring equinox and (2) melatonin treatment would result in an earlier release from estradiol negative feedback and advance the onset of puberty in gilts born around the spring equinox. We first determined the optimal number of estradiol implants required to monitor the release from estradiol negative feedback in ovariectomised gilts. Secondly we determined whether melatonin implants altered negative feedback in 4 cohorts of ovariectomised gilts born between the winter solstice and spring equinox, and in the following year whether melatonin altered the time of the first ovulation in 5 cohorts of intact gilts born between the winter solstice and spring equinox. Plasma LH and FSH increased between 126 and 210d of age (P<0.001) in each cohort (season), but there was no effect of cohort, melatonin treatment or interactions (P>0.05). Age at first detection of elevated plasma progesterone in untreated, intact gilts decreased across the 4 cohorts (P<0.05). Melatonin treatment of intact gilts failed to advance the age of puberty irrespective of their season of birth (P>0.05). In conclusion, while we confirmed that estradiol sensitivity is decreased as gilts age, we failed to demonstrate any effects of season or melatonin on estradiol feedback or melatonin on puberty.

Human and bovine viruses in the Milwaukee River watershed: hydrologically relevant representation and relations with environmental variables.

To examine the occurrence, hydrologic variability, and seasonal variability of human and bovine viruses in surface water, three stream locations were monitored in the Milwaukee River watershed in Wisconsin, USA, from February 2007 through June 2008. Monitoring sites included an urban subwatershed, a rural subwatershed, and the Milwaukee River at the mouth. To collect samples that characterize variability throughout changing hydrologic periods, a process control system was developed for unattended, large-volume (56-2800 L) filtration over extended durations. This system provided flow-weighted mean concentrations during runoff and extended (24-h) low-flow periods. Human viruses and bovine viruses were detected by real-time qPCR in 49% and 41% of samples (n=63), respectively. All human viruses analyzed were detected at least once including adenovirus (40% of samples), GI norovirus (10%), enterovirus (8%), rotavirus (6%), GII norovirus (1.6%) and hepatitis A virus (1.6%). Three of seven bovine viruses analyzed were detected including bovine polyomavirus (32%), bovine rotavirus (19%), and bovine viral diarrhea virus type 1 (5%). Human viruses were present in 63% of runoff samples resulting from precipitation and snowmelt, and 20% of low-flow samples. Maximum human virus concentrations exceeded 300 genomic copies/L. Bovine viruses were present in 46% of runoff samples resulting from precipitation and snowmelt and 14% of low-flow samples. The maximum bovine virus concentration was 11 genomic copies/L. Statistical modeling indicated that stream flow, precipitation, and season explained the variability of human viruses in the watershed, and hydrologic condition (runoff event or low-flow) and season explained the variability of the sum of human and bovine viruses; however, no model was identified that could explain the variability of bovine viruses alone. Understanding the factors that affect virus fate and transport in rivers will aid watershed management for minimizing human exposure and disease transmission.

Lactation estrus induction in multi- and primiparous sows in an Australian commercial pork production system.

This study evaluated the effect of full physical boar exposure and split weaning on the incidence of lactation estrus within a large commercial piggery. A total of 299 multiparous (MP; parity 2.5 ± 0.03) and 303 primiparous (PP) sows of Large White × Duroc × Landrace genetics were individually housed in conventional farrowing crates from 1 wk before expected farrowing until weaning on Day 30.7 ± 0.05 postparturition. Before shed entry, sows were allocated randomly within parity to receive either boar exposure (BE; n = 454) or no BE (No BE; n = 149). Sows assigned to receive BE were then allocated to 1 of 2 litter size treatments: litter size unchanged (BE; n = 302) or BE and the litter permanently reduced (split weaned) to 7 piglets (BESPW7; n = 152) on Day 18 of lactation. From Day 18 of lactation until weaning, sows in both BE treatments were taken daily to a detection mating area where they received 15 min of full physical BE and were artificially inseminated at the first observed estrus. Providing sows with BE increased the incidence of lactation estrus, with a further increase observed when litter size was reduced to 7 piglets (16% No BE vs. 62% BE and 75% BESPW7; P < 0.05). Multiparous sows exhibited a greater incidence of lactation estrus than PP sows irrespective of treatment (81 compared to 52%, respectively; P < 0.05). Both MP and PP sows exhibited an increased incidence of lactation estrus when a portion of the litter was removed (MP: 76 vs. 89% and PP: 47 vs. 61%; P < 0.05). Farrowing rates were higher in BE MP sows mated postweaning and all BESPW7 sows mated postweaning when compared to their counterparts mated in lactation (P < 0.05). Percentage live weight loss over the course of lactation was greatest for sows in the No BE compared to the BE and BESPW7 treatments (7.7% ± 0.5 vs. 5.4% ± 0.3 and 4.5% ± 0.4, respectively; P < 0.05). Between Day 17 and weaning, piglets suckling sows in the BESPW7 treatment had a higher average weight gain than piglets suckling sows with a full litter (3.5 ± 0.06 vs. 3.1 ± 0.05 kg; P < 0.05). In conclusion these data suggest that providing MP sows with BE is effective at stimulating a synchronous lactation estrus while PP sows require, in addition to BE, a reduction in suckled litter size to 7 piglets.

Supplementing sow gestation diets with betaine during summer increases litter size of sows with greater numbers of parities.

Conducted during the Australian summer, this experiment evaluated the reproductive performance of sows receiving a diet supplemented with betaine, a potent organic osmolyte and methyl donor. Large White/Landrace/Duroc sows (n=450) ranging in parity from 1 to 7 (parity 2.9 ± 0.10, mean ± SEM), and mated between the 11th of January and 11th February were used. The treatments compared the effects of two gestation diets (standard (Stand) compared to betaine (Bet) supplemented) and two parity groups (parities one and two (P1/2) versus parity three and greater (P3+) on pregnancy outcomes and litter size. The betaine diet was fed from d 3 ± 1 post-mating until farrowing, with betaine content of the diet altered during gestation to ensure a daily intake of 7.6-9.0 g/sow. Liveweight (LW) and LW gain were unaffected by gestation diet; however, on d 1 of lactation P2 backfat (P2) tended (P=0.07) to be greater for standard compared to betaine fed sows (22.5 ± 0.42 compared to 21.5 ± 0.42 mm). P2, LW and LW gain were greater (P<0.05) for P3+ compared to P1/2 sows. Sow farrowing rate (0.79) was unaffected by gestation diet. Total litter size was greater (P<0.05) for Bet3+ (13.6 ± 0.35) sows compared to Stand3+ (12.1 ± 0.34), BetP1/2 (12.1 ± 0.36) and StandP1/2 (12.3 ± 0.38) sows. In conclusion, this is the first study to demonstrate that gestational betaine supplementation during summer increased litter size of sows with greater numbers of parities.

Nutritional restriction of pre-pubertal liveweight gain impairs ovarian follicle growth and oocyte developmental competence of replacement gilts.

The objective was to investigate the effects of moderate restriction of pre- and peri-pubertal liveweight gain on ovarian development and oocyte meiotic competence. At 70 d of age, and 27.7 ± 0.4 kg liveweight (LW), 64 Large White/Landrace crossbred gilts were allocated to two treatment groups (n = 32 gilts/treatment); one group was fed to attain a LW of 70 kg at 161 d of age (LIGHT), while the other group was fed to reach 100 kg LW (HEAVY). At 161 d of age, half of the gilts in each group (n = 16) were fed to gain LW at 0.5 kg/d (LOW), while the remaining half (n = 16) were fed to gain LW at 1.0 kg/d (HIGH) between 161 and 175 d of age, at which point they were killed and ovaries collected. For each gilt, surface antral follicles were counted and aspirated according to three size categories: 1-2.9 mm (small); 3-6 mm (medium); and > 6 mm (large). Follicles were pooled for each size class and treatment. Cumulus-oocyte-complexes (COC) recovered from small and medium follicles were matured in vitro (IVM) for 44 to 46 h, and meiotic maturation assessed. There was an effect of treatment (LIGHT versus HEAVY) on the number of medium sized follicles: 25.1 ± 2.59 versus 34.3 ± 2.60 (P < 0.05). The ovaries of LOW gilts had more small follicles and fewer medium follicles compared to those of HIGH gilts: 92.8 ± 8.35 versus 59.8 ± 5.24, and 25.1 ± 2.59 versus 32.5 ± 2.86 (P < 0.05). Target LW at 161 d did not affect meiotic progression of oocytes. However, LOW compared to HIGH LW gain between 161 and 175 d resulted in fewer oocytes reaching MII (0.40 versus 0.54; P < 0.05). In conclusion, moderately restricting feed intake impaired follicle growth beyond 3 mm and reduced oocyte meiotic competence. Further, although a carry-over effect of long-term feed restriction on follicle growth was evident, acute changes in feed intake during the 14 d prior to ovary collection had the greatest effect on oocyte nuclear maturation in vitro.

Ractopamine hydrochloride improves growth performance and carcass composition in immunocastrated boars, intact boars, and gilts.

The beta-agonist ractopamine is a dietary ingredient that improves growth and increases the lean mass with little change in fat mass in gilts and barrows. Limited data in boars indicate that dietary ractopamine may increase lean tissue and decrease fat deposition, whereas there are no data for immunocastrated boars. The aims of this investigation were 1) to assess whether the growth performance of all sexes could be maintained over 31 d by using a step-up dietary ractopamine feeding program of 5 mg/kg of ractopamine for the first 14 d, then increasing the dose to 10 mg/kg for a further 17 d, and 2) to determine if dietary ractopamine would increase lean mass in all sexes and decrease fat mass in boars and immunocastrated boars. The study involved 286 pigs randomized and proportionally allocated by breed into 24 groups of 11 or 12 pigs at 17 wk of age, with equal groups of boars, immunocastrated boars, and gilts. Dietary ractopamine decreased (P = 0.005) ADFI during the first 2 wk, particularly in the intact and immunocastrated boars, with the reduction in ADFI being maintained in the immunocastrated boars after the increment in dietary ractopamine. Daily BW gain was not altered by dietary ractopamine during the first 2 wk, but was increased (P < 0.001) after the increment in dietary ractopamine. Dietary ractopamine decreased (P < or = 0.033) feed conversion ratio in all sexes with the response being greater after the increase in dietary ractopamine. Carcass weight was increased (P < 0.001) by dietary ractopamine in all sexes, whereas back fat tended (P = 0.076) to be reduced in the immunocastrated boars. Dietary ractopamine increased (P = 0.018) lean tissue mass by 4.0, 4.8, and 6.5 kg in the intact boars, gilts, and immunocastrated boars, respectively. In the entire and immunocastrated boars, the increase in lean tissue was accompanied with a decrease (P = 0.004) in fat mass. There was little effect of dietary ractopamine on fat mass in gilts. However, carcass percent fat was decreased (P = 0.004) and percent lean increased (P = 0.006) in all sexes. Immunocastration caused a decrease in lean tissue mass and an increase in fat mass and an increase in ADFI in the last one-half of the study. Dietary ractopamine may decrease fat mass in intact and immunocastrated boars and offers an excellent means of maximizing the effects of immunocastration and minimizing the increase in fat mass sometimes observed in immunocastrated boars.

The effects of PG600 and boar exposure on oestrus detection and potential litter size following mating at either the induced (pubertal) or second oestrus.

Within gilt pools, incidences of delayed puberty attainment, failure to exhibit regular oestrous cycles and low first litter size are often high. Boar exposure is an effective method of accelerating puberty; however, the timing of gilt response can vary greatly. Although, PG600 (400 IU of PMSG and 200 IU of hCG; Intervet) can induce a rapid and synchronous ovulatory response, thus providing an alternative to boar contact, the quality of the response is often variable. This study compared the effect of PG600, either alone (NBC) or in conjunction with boar exposure (BC), on puberty attainment and maintenance of oestrous cyclicity. The effects of first mating these gilts at the hormonally induced (pubertal) or second oestrus on ovulation rate and early embryo survival were also studied. Eighty Large White cross terminal (Duroc) line gilts were used in this study. The study was conducted in two blocks, with 10 gilts allocated to each of the four treatments in each block. Gilts were artificially inseminated at the allocated oestrus, with the reproductive tracts collected at 26.5+/-0.29 days after first mating (mean+/-S.E.M.), and the number of corpora lutea and viable embryos recorded. Mean days-to-puberty was significantly reduced (P<0.05) when gilts received both PG600 and boar exposure as opposed to PG600 alone (5.7+/-0.15 versus 6.9+/-0.37 days; P<0.01). The proportion of gilts exhibiting an ovulatory response to PG600 was similar for the BC and NBC treatment groups (0.88 and 0.84); however, the proportion of gilts exhibiting visible signs of oestrus in response to PG600 was significantly higher for the BC compared to the NBC treatment groups (0.81 versus 0.49; P<0.05). Boar contact resulted in a numerical, but not significant, increase in the proportion of gilts exhibited a second oestrus (1.00 versus 0.76). There was no significant effect of boar contact on ovulation rate, embryo number or survival. Although ovulation rate was unaffected by oestrus at mating, embryo number was significantly increased (P<0.05) following mating at the second compared to the first oestrus (11.2+/-0.96 versus 7.8+/-1.17). In conclusion, the current data indicate that the timing of puberty attainment and oestrus detection are significantly improved when PG600 treated gilts receive full boar contact. Further, it is evident that mating gilts at their second as opposed to the hormonally induced oestrus significantly increases embryo number at day 26 post-mating.

Mixing gilts in early pregnancy does not affect embryo survival.

There is general acceptance that mixing sows during the first 3 weeks of gestation is detrimental to embryo development and survival. However, there is a paucity of data describing the influence of group housing and remixing during the first 14 days of gestation on pregnancy outcomes. Using 96 purebred maternal (Large White)/terminal (Duroc) line gilts, the current study determined the effects of regrouping, and the timing of regrouping, during the pre-implantation period on embryo mortality. The study was conducted in 2 blocks, with 12 gilts allocated to each of 4 treatments in each block. At 175 days of age, the combination of PG600 and 20 min of daily physical boar contact was used to stimulate puberty, with boar contact resuming 12 days after first detection of oestrus and gilts receiving two artificial inseminations (AIs), 24 h apart, at their second oestrus. After their first AI gilts were allocated to one of four treatment groups (n=12 gilts/treatment). Gilts in one treatment group were housed individually in stalls (STALL). The remaining gilts continued to be housed in their pre-AI groups and were either not remixed (NOMIX), or remixed to form new groups on day 3/4 (RMIXD3/4) or day 8/9 (RMIXD8/9) of gestation (day 0=day of first detection of second oestrus and first insemination). Group-housed gilts were housed in groups of 6, with a space allowance of 2.4 m2/gilt. All gilts were fed once a day (2.2 kg/gilt). Reproductive tracts were collected on day 26.6+/-0.13 of gestation, and the number of corpora lutea (CL) and viable embryos counted. Pregnancy rate was similar across all treatments, averaging 94.5% across the four treatment groups. The number of embryos present on day 26 of gestation was unaffected by housing treatments (P>0.05); gilts in the STALL, NOMIX, RMIXD3/4 and RMIXD8/9 groups possessed 13.2+/-0.67, 12.9+/-0.66, 14.1+/-0.46 and 13.8+/-0.57 embryos, respectively. Similarly, embryo survival rates were 0.91+/-0.04, 0.85+/-0.04, 0.91+/-0.02 and 0.87+/-0.05 for the STALL, NOMIX, RMIXD3.4 and RMIXD8/9 groups, respectively (P>0.05). In conclusion, the current data indicate that individually housing gilts immediately after their first AI does not improve embryo survival. There also appear to be no adverse effects on embryo development or survival when group-housed, mated gilts are remixed during the first 10 days of gestation.

Increasing the age of gilts at first boar contact improves the timing and synchrony of the pubertal response but does not affect potential litter size.

One hundred and ninety-two Large White/Landrace crossbred gilts were used in this study. The study was conducted in two blocks, with 16 gilts allocated to each of six treatments in each block. The treatments compared the effects on puberty attainment of commencing boar exposure at 161, 182 or 203 days of age, and the effect of first mating gilts at either the pubertal or second oestrus on ovulation rate and early embryo survival. Boar contact took place in a detection-mating area (DMA), and consisted of 20 min/day of full contact with a vasectomized boar greater than ten months of age. Gilts were artificially inseminated at the allocated oestrus, with the reproductive tracts collected at 22.8+/-0.4 days after first mating (mean+/-S.E.M.), and the numbers of corpora lutea and viable embryos recorded. The age at which gilts attained puberty increased with the age at which boar exposure commenced. Mean gilt ages at puberty were 179.5+/-1.6, 191.7+/-1.2 and 210.3+/-0.9 days, respectively, when boar contact commenced at 161, 182 and 203 days of age, P<0.01. Mean days-to-puberty was significantly shorter when boar contact began at 182 and 203 days of age compared to 161 days of age (10.4+/-1.2 and 8.3+/-0.9 days versus 18.9+/-1.5 days, respectively, P<0.01). Similarly, commencing boar exposure at 182 or 203 days of age as opposed to 161 days of age significantly increased the proportion of gilts attaining puberty within 10 days of start of boar exposure (0.67 and 0.70 versus 0.24, P<0.01). Mean days-to-puberty and the synchrony of puberty attainment were similar when boar contact commenced at 182 and 203 days of age. There was no significant effect of mating oestrus or age of gilts at mating on ovulation rate, embryo number or embryo survival. In conclusion, the current data indicate that the timing and synchrony of puberty attainment is significantly improved when first boar exposure of gilts is delayed until 182 days of age. Further, it is evident that within the age range investigated, delaying first mating until the second oestrus does not significantly increase either ovulation rate or embryo number at day 20 post-mating.

Integrin activation controls metastasis in human breast cancer.

Metastasis is the primary cause of death in human breast cancer. Metastasis to bone, lungs, liver, and brain involves dissemination of breast cancer cells via the bloodstream and requires adhesion within the vasculature. Blood cell adhesion within the vasculature depends on integrins, a family of transmembrane adhesion receptors, and is regulated by integrin activation. Here we show that integrin alpha v beta 3 supports breast cancer cell attachment under blood flow conditions in an activation-dependent manner. Integrin alpha v beta 3 was found in two distinct functional states in human breast cancer cells. The activated, but not the nonactivated, state supported tumor cell arrest during blood flow through interaction with platelets. Importantly, activated alpha v beta 3 was expressed by freshly isolated metastatic human breast cancer cells and variants of the MDA-MB 435 human breast cancer cell line, derived from mammary fat pad tumors or distant metastases in severe combined immunodeficient mice. Expression of constitutively activated mutant alpha v beta 3(D723R), but not alpha v beta 3(WT), in MDA-MB 435 cells strongly promoted metastasis in the mouse model. Thus breast cancer cells can exhibit a platelet-interactive and metastatic phenotype that is controlled by the activation of integrin alpha v beta 3. Consequently, alterations within tumors that lead to the aberrant control of integrin activation are expected to adversely affect the course of human breast cancer.

The effector loop and prenylation site of R-Ras are involved in the regulation of integrin function.

The closely related small GTP-binding proteins H-Ras and R-Ras have opposing effects on the regulation of integrin cell adhesion receptors. To gain insight into the properties of R-Ras with respect to the regulation of integrin function and interactions with downstream effectors we performed an analysis of R-Ras variants containing mutations in the effector binding domain and C-terminal prenylation site. We found that the activation of the downstream effector PI 3-kinase was sensitive to mutations in the effector binding domain, as was the binding to the effectors, Ral-GDS, Raf-1 and the novel effector Nore1. Furthermore, specific mutations in the effector binding loop and C-terminal prenylation motif impaired the ability of R-Ras to regulate integrin function in CHO cells. However, the ability of the R-Ras effector loop mutants to bind, and activate known effectors did not correlate with their ability to regulate integrin function. Thus, the known R-Ras effectors are not critical for regulating integrin activation, at least in CHO cells. Consequently, these studies provide insight into the structural basis of the interactions between R-Ras and its candidate effectors and suggest the existence of novel mechanisms through which this GTPase could regulate cell adhesion.

Death effector domain protein PEA-15 potentiates Ras activation of extracellular signal receptor-activated kinase by an adhesion-independent mechanism.

PEA-15 is a small, death effector-domain (DED)-containing protein that was recently demonstrated to inhibit tumor necrosis factor-alpha-induced apoptosis and to reverse the inhibition of integrin activation due to H-Ras. This led us to investigate the involvement of PEA-15 in Ras signaling. Surprisingly, PEA-15 activates the extracellular signal receptor-activated kinase (ERK) mitogen-activated protein kinase pathway in a Ras-dependent manner. PEA-15 expression in Chinese hamster ovary cells resulted in an increased mitogen-activated protein kinase kinase and ERK activity. Furthermore, PEA-15 expression leads to an increase in Ras guanosine 5'-triphosphate loading. PEA-15 bypasses the anchorage dependence of ERK activation. Finally, the effects of PEA-15 on integrin signaling are separate from those on ERK activation. Heretofore, all known DEDs functioned in the regulation of apoptosis. In contrast, the DED of PEA-15 is essential for its capacity to activate ERK. The ability of PEA-15 to simultaneously inhibit apoptosis and potentiate Ras-to-Erk signaling may be of importance for oncogenic processes.

Neuroprotective strategies for basal ganglia degeneration: Parkinson's and Huntington's diseases.

There are three main mechanisms of neuronal cell death which may act separately or cooperatively to cause neurodegeneration. This lethal triplet of metabolic compromise, excitotoxicity, and oxidative stress causes neuronal cell death that is both necrotic and apoptotic in nature. Aspects of each of these three mechanisms are believed to play a role in the neurodegeneration that occurs in both Parkinson's and Huntington's diseases. Strategies to rescue or protect injured neurons usually involve promoting neuronal growth and function or interfering with neurotoxic processes. Considerable research has been done on testing a large array of neuroprotective agents using animal models which mimic these disorders. Some of these approaches have progressed to the clinical arena. Here, we review neuroprotective strategies which have been found to successfully ameliorate the neurodegeneration associated with Parkinson's and Huntington's diseases. First, we will give an overview of the mechanisms of cell death and the background of Parkinson's and Huntington's diseases. Then we will elaborate on a range of neuroprotective strategies, including neurotrophic factors, anti-excitotoxins, antioxidants, bioenergetic supplements, anti-apoptotics, immunosuppressants, and cell transplantation techniques. Most of these approaches hold promise as potential therapies in the treatment of these disorders.

A note on the effects of contact frequency and time of day of boar exposure on the efficacy of the boar effect.

Sixty-four Large White/Landrace crossbred gilts were used in this study, 16 gilts being allocated to each of four treatments to compare the effects on puberty attainment of exposure to boar contact either 0, 1 or 2 times daily. The once-daily exposure occurred in either the morning or the afternoon (AM vs. PM). Treatments were of 20-min duration starting at a mean gilt age of 160 days and continuing for 60 days. Boar exposure significantly increased the proportion of gilts attaining puberty within 60 days of the commencement of treatments (P < 0.05) compared with gilts not receiving boar contact. Gilts receiving boar exposure twice daily attained puberty significantly earlier than did gilts in the two treatment groups (AM and PM, respectively) given a single daily boar exposure period (mean gilt ages at puberty 176.4 vs. 192.7 and 189.2 days of age, respectively, P < 0.05). It is concluded that (a) twice-daily boar contact enhances the efficacy of the boar effect in gilts above that seen with a single daily boar exposure period and (b) this enhanced response of the gilt is due to the frequency of boar contact and not to the time of day at which the contact occurs.

The IGF-I amino-terminal tripeptide glycine-proline-glutamate (GPE) is neuroprotective to striatum in the quinolinic acid lesion animal model of Huntington's disease.

Huntington's disease is an incurable genetic neurological disorder characterized by the relatively selective degeneration of the striatum. Lesioning of the striatum in rodents using the excitatory amino acid agonist, quinolinic acid (QA), effectively mimics the human neuropathology seen in Huntington's disease. Using this animal model of Huntington's disease, we investigated the ability of the insulin-like growth factor-I (IGF-I) amino-terminal tripeptide glycine-proline-glutamate (GPE) to protect striatal neurons from degeneration. Adult rats received a single unilateral intrastriatal injection of QA (100 nmol) and then daily injection of either vehicle or GPE (0.3 microgram/microliter/day) into the striatum for 7 days. QA at this dose resulted in a partial lesioning of the striatum after 7 days to approximately 50% of cells of unlesioned levels in vehicle-treated animals. The major striatal neuronal phenotype, GABAergic projection neurons, were identified by immunocytochemical labeling of either glutamate decarboxylase 67 (GAD(67)) or the calcium binding protein calbindin in alternate sections. Treatment with GPE for 7 days reversed the loss in projection neurons when assessed by counts of calbindin-stained cells; however, these rescued cells did not regain immunologically detectable levels of GAD(67). GPE also significantly reversed the phenotypic degeneration of cholinergic interneurons identified by immunolabeling for choline acetyltransferase (ChAT) and NADPH diaphorase interneurons identified histochemically. GPE treatment failed to rescue the calcium binding protein interneuron populations of parvalbumin and calretinin neurons. These findings reveal that exogenous administration of GPE selectively prevents excitotoxin induced phenotypic degeneration of striatal projection neurons and cholinergic and NADPH diaphorase interneurons in an animal model of Huntington's disease.