Liver development is restored by blastocyst complementation of HHEX knockout in mice and pigs.

BACKGROUND: There are over 17,000 patients in the US waiting to receive liver transplants, and these numbers are increasing dramatically. Significant effort is being made to obtain functional hepatocytes and liver tissue that can for therapeutic use in patients. Blastocyst complementation is a challenging, innovative technology that could fundamentally change the future of organ transplantation. It requires the knockout (KO) of genes essential for cell or organ development in early stage host embryos followed by injection of donor pluripotent stem cells (PSCs) into host blastocysts to generate chimeric offspring in which progeny of the donor cells populate the open niche to develop functional tissues and organs. METHODS: The HHEX gene is necessary for proper liver development. We engineered loss of HHEX gene expression in early mouse and pig embryos and performed intraspecies blastocyst complementation of HHEX KO embryos with eGFP-labeled PSCs in order to rescue the loss of liver development. RESULTS: Loss of HHEX gene expression resulted in embryonic lethality at day 10.5 in mice and produced characteristics of lethality at day 18 in pigs, with absence of liver tissue in both species. Analyses of mouse and pig HHEX KO fetuses confirmed significant loss of liver-specific gene and protein expression. Intraspecies blastocyst complementation restored liver formation and liver-specific proteins in both mouse and pig. Livers in complemented chimeric fetuses in both species were comprised of eGFP-labeled donor-derived cells and survived beyond the previously observed time of HHEX KO embryonic lethality. CONCLUSIONS: This work demonstrates that loss of liver development in the HHEX KO can be rescued via blastocyst complementation in both mice and pigs. This complementation strategy is the first step towards generating interspecies chimeras for the goal of producing human liver cells, tissues, and potentially complete organs for clinical transplantation.

Teaching emotional and social competencies: Efficacy of a work readiness program designed for vulnerable youth.

BACKGROUND: As emotional and social competency training proliferates within a work readiness context, concerns remain regarding their efficacy. Data on these programs tends to be scarce and outcome objectives are often poorly defined. OBJECTIVE: Authors developed and tested a work readiness emotional and social competency program specifically designed for at-risk young adults, tailored with best practices in mind. METHOD: 84 clients of a community organization that provides employment support to young adults with disabilities (48 men and 36 women) with a mean age of 28.17 years (SD = 11.64) completed measures of emotional intelligence and alexithymia on either side of the 4-week intervention. RESULTS: Men's interpersonal scores and women's adaptability scores showed significant improvement across the intervention. In addition, women's scores in both identifying and describing feelings improved significantly, as did men's scores in describing feelings. CONCLUSIONS: Within the context of work readiness, participants in an intervention to improve emotional and social competencies can see key improvements to competencies linked to occupational attainment.

The development of a noninvasive behavioral test for assessment of goat-human interactions.

The objective was to develop a protocol for a noninvasive behavioral test to assess goat-human interactions. Boer goats (n = 45) were housed in groups of 3 at the K-State Sheep and Meat Goat Unit. A 3-min human approach test (HAT) was conducted after their first- and seventh-handling experience (i.e., moved by group through the chute and scale). Video footage was analyzed using specialized software (Observer 11.5 XT, Leesburg, VA, USA). The ethogram included three main categories (mutually exclusive within category): 1) spatial (close, middle, far); 2) orientation (facing vs. turned-away); and 3) structural (lie, stand, and nutritive and nonnutritive oral behaviors). The durations of these behavior outcomes were converted to percent, and then applied to a weighted formula to create an approach index (AI). This index placed behaviors on a 0 to 100% scale (0 = lying in back of the pen; 100 = closest to the human, performing oral behaviors). The Proc Univariate (SAS v.9.3, SAS Inst. Inc., Cary, NC, USA) was used to evaluate descriptive statistics and calculate sample size for future projects from all behavior outcomes. In addition, each goat AI categorized as great approach (GA; ≥75% quartile), moderate approach (MA; 25% to 75% quartiles), or least approach (LA; ≤ 25%) and the 1st vs. 7th handling experience was evaluated using Chi-square (χ 2) analysis. The CV% was low for the AI outcome; therefore, less animals are needed if the AI is used rather than other behavior outcomes used. After the first-handling experience, there were less GA-goats than what was expected from the χ 2 analysis (χ 2 = 17.6; P ≤ 0.01; residual = 0.26; expected = 24.5). After the seventh-handling experience, there were more MA-goats than expected (χ 2 = 17.6; P ≤ 0.01; residual = 1.92; expected = 52.0). Repeated handling appeared to moderately improve goat approach to humans, which indicates that this protocol is a promising behavioral test to assess welfare of goats.

Impact of including distillers' dried grains with solubles at expense of soybean meal on Boer-influenced goat growth performance.

The objective of this project was to evaluate the efficacy of distillers' dried grains (DDGS) as a replacement for soybean meal (SBM) in a diet for Boer-influenced goats. Because diets were isonitrogenous and isocaloric, it was hypothesized that the replacement of SBM with DDGS would reduce diet costs without affecting growth performance. To test this hypothesis, forty-eight Boer-influenced goats (28.2 ± 0.96 kg BW; approximately 90 d of age) were allocated to four treatments in a completely random design in a climate-controlled (13°C) facility of the Kansas State University Sheep and Meat Goat Center. Goats were stratified by BW into 16 pens (four pens per treatment; three goats per pen) for a 42-d finishing study. The main effect concentration of DDGS (0%, 10%, 20%, or 30% DM basis) which resulted in dietary treatments of the following: 1) 0% SBM replaced by DDGS (0DDGS); 2) 33% SBM replaced by DDGS (10DDGS); 3) 66% SBM replaced by DDGS (20DDGS); and 4) 100% SBM replaced by DDGS (30DDGS). Goats remained on a self-feeder with continuous access to their respective pelleted, complete dietary treatments, and clean, fresh water. Two-day weights were taken at the beginning and end of the trial, with BW, ADG, ADFI, and G:F measured and calculated every 7 d. Beginning BW were similar across all treatments (P = 0.99). The inclusion of DDGS had no effect on BW or ADFI from days 0 to 42 (P ≥ 0.15). Both ADG (P = 0.04) and G:F (P = 0.001) increased linearly with increasing DDGS inclusion, with goats fed diets containing DDGS having 33% greater (P = 0.05) ADG than those fed diets without DDGS. Ultimately, these results confirm the hypothesis that DDGS can be used to replace SBM in the diet. At current commodity prices, this leads to a 17% savings in diet cost. This novel research shows similar improved performance measures to lambs and cattle when feeding DDGS.

Effects of dried distiller's grains and lasalocid inclusion on feedlot lamb growth, carcass traits, nutrient digestibility, ruminal fluid volatile fatty acid concentrations, and ruminal hydrogen sulfide concentration.

Our hypothesis was that increasing the inclusion level of dried distiller's grains with solubles (DDGS) to feedlot lambs would increase growth and the inclusion of lasalocid (LAS; Bovatec, Alpharma, LLC, Bridgewater, NJ) would increase ADG and G:F, while not affecting digestibility, ruminal VFA concentration, and ruminal pH. Furthermore, we hypothesized that rations containing LAS and higher levels of DDGS would cause increased ruminal hydrogen sulfide gas (HS) concentrations. Two hundred forty crossbred (Suffolk × Rambouillet) lambs (31.9 ± 5.87 kg BW; approximately 90 d of age) were allocated to 6 treatments in a completely randomized design with a 3 × 2 factorial arrangement of treatments. Lambs were placed into 24 feedlot pens (4 pens/treatment; 10 lambs/pen) for a 111 d finishing study. Main effects included concentration of DDGS (0, 15, or 30% DM basis) and inclusion of LAS (0 or 22.05 g/metric ton LAS) resulting in treatments of: 1) 0% DDGS without LAS (0DDGS-NL), 2) 0% DDGS with LAS (0DDGS-L), 3) 15% DDGS without LAS (15DDGS-NL), 4) 15% DDGS with LAS (15DDGS-L), 5) 30% DDGS without LAS (30DDGS-NL), and 6) 30% DDGS with LAS (30DDGS-L). Two-day weights were taken at the beginning and end of the experiment. Two-hundred-eighteen lambs (64.8 ± 7.99 kg BW) were slaughtered on d 112 at a commercial abattoir and carcass data collected. The inclusion of LAS increased ( ≤ 0.02) final BW, ADG, G:F, and HCW. As DDGS in the ration increased to 30%, DMI decreased linearly ( = 0.03) while G:F increased linearly ( = 0.03). A second study was conducted utilizing the same treatments to evaluate N and S balance, ruminal VFA and H2S concentration, and ruminal pH in 24 crossbred wethers (Suffolk × Rambouillet; 41.2 ± 12.23 kg BW). Daily urinary sulfur excretion and ruminal H2S concentration were linearly increased ( < 0.001) as DDGS increased in the ration. Total ruminal VFA concentration linearly decreased ( = 0.002) as DDGS increased in the ration. The inclusion of LAS increased ( = 0.02) ruminal pH. The results confirm our hypothesis that LAS increased overall growth and increasing DDGS increased ruminal HS concentration but did not influence growth. We reject the hypothesis that the combined effects of LAS and DDGS would have no effect on rumen pH and VFA concentrations.

Impacts of supplemental arginine on the reproductive performance of fall lambing ewes.

In sheep, embryonic and fetal death during pregnancy can account for 25% to 50% of the total number of corpora lutea (and thus potential embryos). The objective of this study was to determine the effects of injectable and oral Arg supplementation provided for 14 d postbreeding on the reproductive performance of naturally stimulated fall lambing ewes. Rambouillet ewes ( = 210) were exposed to rams equipped with marking harnesses to induce cyclicity in April 2012. Upon estrus detection (d 0) ewes were randomly assigned, in a completely random design, to 1 of 6 treatments for a 14-d treatment period: injectable saline (CON; = 25), injectable Ala (IVALA; = 20), injectable Arg (IVARG; = 23), oral rumen-protected Arg (RPARG; = 20), oral fish meal (FM; = 24), or oral soybean meal (SBM; = 23). Daily treatments, except CON, IVALA, and SBM, were formulated to provide supplemental Arg at 30 mg·kg BW·d and were provided at 0800 h daily. Ewes receiving injectable treatments were provided 454 g corn/d postinjection, whereas ewes receiving oral supplements were provided a ground ration of their respective treatments with corn individually at 0800 h daily. Plasma and serum samples were collected on d 0, 2, 4, 6, 8, 10, 12, and 14 from 12 ewes per treatment to evaluate plasma progesterone and serum AA concentrations. At lambing, birth weight, birth type, and sex were recorded. Weaning weights were recorded when the average age of lambs was 85 d. No differences ( ≥ 0.39) were detected for pregnancy, prolificacy, and lambing rates or lamb birth weights among treatments. However, litter weaning weight tended to be greater ( = 0.06) and weaning rates were greater ( = 0.05) in Arg-injected ewes (1.09, 0.95, 1.29, 0.72, 1.00, and 0.86, respectively). Plasma progesterone and serum Arg concentrations showed a treatment and day effect ( < 0.001), but no treatment × day interaction ( ≥ 0.99) was observed. In contrast to previous research, supplemental Arg during the first 14 d of pregnancy did not improve pregnancy or lambing rates; however, IVARG did positively impact weaning rates.

Maternal environment and placental vascularization in small ruminants.

Uteroplacental development is a crucial step facilitating conceptus growth. Normal placental development comprises extensive placental angiogenesis to support fetoplacental transport, meeting the metabolic demands of the fetus. Compromised pregnancies due to maternal stressors such as over or undernutrition, maternal age or parity, altered body mass index, or genetic background result in altered vascular development of the placenta. This negatively affects placental growth and placental function and ultimately results in poor pregnancy outcomes. Nonetheless, the placenta acts as a sensor to the maternal stressors and undergoes modifications, which some have termed placental programming, to ensure healthy development of the conceptus. Sex steroid hormones such as estradiol-17ß and progesterone, chemokines such as chemokine ligand 12, and angiogenic/vasoactive factors such as vascular endothelial growth factors, placental growth factor, angiopoietins, and nitric oxide regulate uteroplacental development and hence are often used as therapeutic targets to rescue compromised pregnancies. Interestingly, the presence of sex steroid receptors has been identified in the fetal membranes (developing fetal placenta). Environmental steroid mimetics known as endocrine disrupting compounds disrupt conceptus development and lead to transgenerational impairments by epigenetic modification of placental gene expression, which is another area deserving intense research efforts. This review attempts to summarize current knowledge concerning intrinsic and extrinsic factors affecting selected reproductive functions with the emphasis on placental development.

Bone-marrow-derived mesenchymal stem cells attenuate cognitive deficits in an endothelin-1 rat model of stroke.

PURPOSE: Stroke is the third leading cause of death and permanent disability in the United States, often producing long-term cognitive impairments, which are not easily recapitulated in animal models. The goals of this study were to assess whether: (1) the endothelin-1 (ET-1) model of chronic stroke produced discernable cognitive deficits; (2) a spatial operant reversal task (SORT) would accurately measure memory deficits in this model; and (3) bone-marrow-derived mesenchymal stem cells (BMMSCs) could reduce any observed deficits. METHODS: Rats were given unilateral intracerebral injections of vehicle or ET-1, a stroke-inducing agent, near the middle cerebral artery. Seven days later, they were given intrastriatal injections of BMMSCs or vehicle, near the ischemic penumbra. The cognitive abilities of the rats were assessed on a novel SORT, which was designed to efficiently distinguish cognitive deficits from potential motoric confounds. RESULTS: Rats given ET-1 had significantly more cognitive errors at six weeks post-stroke on the SORT, and that these deficits were attenuated by BMMSC transplants. CONCLUSIONS: These findings indicate that: (1) the ET-1 model produces chronic cognitive deficits; (2) the SORT efficiently measures cognitive deficits that are not confounded by motoric impairment; and (3) BMMSCs may be a viable treatment for stroke-induced cognitive dysfunction.

Perineal body length as a risk factor for ultrasound-diagnosed anal sphincter tear at first delivery.

INTRODUCTION AND HYPOTHESIS: Shortened perineal body (PB) is associated with an increased risk of ultrasound-detected obstetric anal sphincter tear. The objective was to determine if shortened perineal body length (<3 cm) is a risk factor for ultrasound-detected anal sphincter tear at first delivery. METHODS: Pregnant nulliparous women were recruited over 18 months. At 35-37 weeks' gestation and 6 weeks' postpartum perineal body length (PB) was measured and subjects completed quality of life questionnaires. Primary outcome was ultrasound-diagnosed anal sphincter tear at 6 weeks postpartum. Secondary outcomes were also assessed. A priori power analysis determined that 70 subjects were needed to detect a difference in anal sphincter tear based on a PB cut-off of 3 cm. RESULTS: Seventy-three subjects completed the study. Mode of delivery was 69.9% spontaneous vaginal, 15.1% operative vaginal, and 15.1% labored cesarean. There were 25 anal sphincter abnormalities (34.2%) seen on ultrasound: 11 (15.1%) internal or external sphincter tears, 3 (4.1%) internal sphincter atrophy, 6 (8.2%) external sphincter thinning, and 7 (9.6%) external sphincter scarring. Only the 11 sphincter tears qualified as abnormal for the primary outcome. In the vaginal delivery group 16.4% (10 out of 61) had a sphincter tear, compared with 8.3% (1 out of 12) in the labored cesarean group (p = 0.68). Women with PB < 3 had a significantly higher rate of ultrasound-diagnosed anal sphincter tear (40.0% vs 11.1%, p = 0.038). When comparing women with and without sphincter tear, there was a significant difference in mean antepartum PB (3.1 vs 3.7 cm, p = 0.043). CONCLUSIONS: A shortened perineal body length in primiparous women is associated with an increased risk of anal sphincter tear at the time of first delivery.

Intragastric self-infusion of ethanol in high- and low-drinking mouse genotypes after passive ethanol exposure.

Two experiments examined the effect of 5 days of passive exposure to ethanol (or water) on later self-infusion of ethanol or water via surgically implanted intragastric (IG) catheters in mouse genotypes previously shown to drink high (C57BL/6J, HAP2) or low (DBA/2J, LAP2) amounts of ethanol in home-cage continuous-access two-bottle choice procedures. Intragastric ethanol self-infusion was affected by both genotype and a history of passive ethanol exposure, with greater intakes in the high-drinking genotypes and in groups that received passive exposure to ethanol. Passive ethanol exposure also increased preference for the flavor that signaled ethanol infusion (S+), eliminating genetic differences in this measure. The increases in ethanol intake and S+ preference induced by ethanol exposure might have been mediated jointly by development of tolerance to aversive post-absorptive ethanol effects and negative reinforcement because of alleviation of withdrawal. Bout analyses indicated that ethanol exposure increased ethanol self-infusion by increasing the total number of daily bouts rather than by increasing bout size. These analyses also showed that DBA/2J mice infused larger ethanol bouts and a greater percentage of their total intakes in large bouts than C57BL/6J mice. Overall, these studies suggest that the IG self-infusion procedure is a potentially useful new tool for studying genetic and environmental influences on excessive ethanol intake and preference in mice.

Insulin secretagogues, sulfonylurea receptors and K(ATP) channels.

ATP-sensitive K+ channels, termed K(ATP) channels, provide a link between cellular metabolism and membrane electrical activity in a variety of tissues. Channel isoforms have been identified and are targets for compounds that both stimulate and inhibit their activity resulting in membrane hyperpolarization and depolarization, respectively. Examples include relaxation of vascular smooth muscle and stimulation of insulin secretion. This article reviews the cloning, molecular biology, and structure of K(ATP) channels, with particular focus on the SUR1/K(IR)6.2 neuroendocrine channels that are important for the regulation of insulin secretion. We integrate the extensive pharmacologic structure-activity-relationship data on these channels, which defines a bipartite drug binding pocket in the SUR (sulfonylurea receptor), with recent structure-function studies that identify domains of SUR and K(IR)6.2, the channel pore, which are critical for channel assembly, for gating, and for the ligand-receptor interactions that modulate channel activity. The atomic structure of a sulfonylurea in a protein pocket is used to develop insight into the recognition of these compounds. A homology model of K(ATP) channels, based on VC-MsbA, another member of the ABC protein family, is described and used to position amino acids important for the action of channel openers and blockers within the core of SUR. The model has a central chamber which could serve as a multifaceted binding pocket.

A novel human leucocyte antigen-DRB1 genotyping method based on multiplex primer extension reactions.

We have developed and validated a semi-automated fluorescent method of genotyping human leucocyte antigen (HLA)-DRB1 alleles, HLA-DRB1*01-16, by multiplex primer extension reactions. This method is based on the extension of a primer that anneals immediately adjacent to the single-nucleotide polymorphism with fluorescent dideoxynucleotide triphosphates (minisequencing), followed by analysis on an ABI Prism 3700 capillary electrophoresis instrument. The validity of the method was confirmed by genotyping 261 individuals using both this method and polymerase chain reaction with sequence-specific primer (PCR-SSP) or sequencing and by demonstrating Mendelian inheritance of HLA-DRB1 alleles in families. Our method provides a rapid means of performing high-throughput HLA-DRB1 genotyping using only two PCR reactions followed by four multiplex primer extension reactions and PCR-SSP for some allele groups. In this article, we describe the method and discuss its advantages and limitations.

Joint acceleration during gait in relation to age.

Forty-four male and female subjects with no history of falling and whose ages ranged from 10 to 68 years participated in a series of experiments to assess movement at the joints during gait while walking in a straight line, in pivot turns and in turns of 0.33 and 0.66 m diameter. Acceleration at the joints in the forward and side-to-side direction was measured by dual-axis accelerometers placed at the ankles, knees, hips, shoulders, and on the head. Eye movement was assessed from electrodes placed on the sides of the eyes. The results of the experiments showed that for people whose age was above 40 years, significant increases in the forward-back and side-to-side movements occurred at all joints and progressively increased with age. By age 60, adverse movement of the joints as much as quadrupled in many subjects when compared to people whose age was 20-30 years. The increase in joint acceleration occurred equally in the front-back and side-to-side planes. The mechanism of the increased joint movement may be due to tendon laxness, peripheral neuropathy or loss of central control of gait due to age. Accelerometry may be a much more sensitive technique to analyze abnormalities in gait than standard video or observational gait analysis. Results are given as mean (SD) unless otherwise stated.

The effect of transforming growth factor beta1 gene polymorphisms in ankylosing spondylitis.

OBJECTIVES: To determine whether genetic polymorphisms in or near the transforming growth factor beta1 (TGFB1) locus were associated with susceptibility to or severity of ankylosing spondylitis (AS). METHODS: Five intragenic single-nucleotide polymorphisms (SNP) and three microsatellite markers flanking the TGFB1 locus were genotyped. Seven hundred and sixty-two individuals from 184 multiplex families were genotyped for the microsatellite markers and two of the promoter SNPs. One thousand and two individuals from 212 English and 170 Finnish families with AS were genotyped for all five intragenic SNPs. A structured questionnaire was used to assess the age of symptom onset, disease duration and disease severity scores, including the BASDAI (Bath Ankylosing Spondylitis Disease Activity Index) and BASFI (Bath Ankylosing Spondylitis Functional Index). RESULTS: A weak association was noted between the rare TGFB1 +1632 T allele and AS in the Finnish population (P = 0.04) and in the combined data set (P = 0.03). No association was noted between any other SNPs or SNP haplotype and AS, even among those families with positive non-parametric linkage scores. The TGFB1 +1632 polymorphism was also associated with a younger age of symptom onset (English population, allele 2 associated with age of onset greater by 4.2 yr, P = 0.05; combined data set, allele 2 associated with age of onset greater by 3.2 yr, P = 0.02). A haplotype of coding region SNPs (TGFB1 +869/+915+1632 alleles 2/1/2) was associated with age of symptom onset in both the English parent-case trios and the combined data set (English data set, haplotype 2/1/2 associated with age of onset greater by 4.9 yr, P = 0.03; combined data set, haplotype 2/1/2 associated with greater age of onset by 4.2 yr, P = 0.006). Weak linkage with AS susceptibility was noted and the peak LOD score was 1.3 at distance 2 cM centromeric to the TGFB1 gene. No other linkage or association was found between quantitative traits and the markers. CONCLUSION: This study suggests that the polymorphisms within the TGFB1 gene play at most a small role in AS and that other genes encoded on chromosome 19 are involved in susceptibility to the disease.

Interleukin 10 polymorphisms in ankylosing spondylitis.

Genetic polymorphisms of the IL10 promoter region have been implicated in many autoimmune diseases, including seronegative spondyloarthropathies. We studied three SNPs (IL10-1087, -824, and -597) and two microsatellites (IL10R and IL10G) lying within the promoter region of IL10 for association with susceptibility to and clinical manifestations of ankylosing spondylitis (AS), a common form of spondyloarthritis. Four hundred and sixty-eight individuals from 182 Finnish families affected with AS were studied. No association between individual IL10 promoter region polymorphisms or marker haplotype was observed with susceptibility to AS, but weak association was noted between the IL10-597 and -824 SNPs and age of disease onset (P=0.01 for each SNP). The IL10.G4 allele was associated with BASFI (corrected for disease duration) (P=0.03). We conclude that IL10 promoter polymorphisms have no significant effect on susceptibility to AS, but may play a minor role in determining age of disease onset and disease severity.

Maintaining the cornea and the general physiological environment in visual neurophysiology experiments.

Neurophysiologists have been investigating the responses of neurons in the visual system for the past half-century using monkeys and cats that are anesthetized and paralyzed, with the non-blinking eyelids open for prolonged periods of time. Impermeable plastic contact lenses have been used to prevent dehydration of the corneal epithelium, which would otherwise occur in minutes. Unfortunately, such lenses rapidly introduce a variety of abnormal states that lead to clouding of the cornea, degradation of the retinal image, and premature termination of the experiment. To extend the viability of such preparations, a new protocol for maintenance of corneal health has been developed. The protocol uses rigid gas permeable contact lenses designed to maximize gas transmission, rigorous sterile methods, and a variety of methods for sustaining and monitoring the overall physiology of the animal. The effectiveness of the protocol was evaluated clinically by ophthalmoscopy before, during, and after the experiments, which lasted 8-10 days. Histopathology and quantitative histology were performed on the corneas following the experiment. Our observations showed that this protocol permits continuous contact lens wear without adversely affecting the corneas. Thus, it is possible to collect data 24 h each day, for the entire duration of the experiment.

An inhibitor of mTOR reduces neoplasia and normalizes p70/S6 kinase activity in Pten+/- mice.

PTEN phosphatase acts as a tumor suppressor by negatively regulating the phosphoinositide 3-kinase (PI3K) signaling pathway. It is unclear which downstream components of this pathway are necessary for oncogenic transformation. In this report we show that transformed cells of PTEN(+/-) mice have elevated levels of phosphorylated Akt and activated p70/S6 kinase associated with an increase in proliferation. Pharmacological inactivation of mTOR/RAFT/FRAP reduced neoplastic proliferation, tumor size, and p70/S6 kinase activity, but did not affect the status of Akt. These data suggest that p70/S6K and possibly other targets of mTOR contribute significantly to tumor development and that inhibition of these proteins may be therapeutic for cancer patients with deranged PI3K signaling.

Deficiency of Pten accelerates mammary oncogenesis in MMTV-Wnt-1 transgenic mice.

BACKGROUND: Germline mutations in the tumor suppressor PTEN predispose human beings to breast cancer, and genetic and epigenetic alterations of PTEN are also detected in sporadic human breast cancer. Germline Pten mutations in mice lead to the development of a variety of tumors, but mammary carcinomas are infrequently found, especially in mice under the age of six months. RESULTS: To better understand the role of PTEN in breast tumor development, we have crossed Pten heterozygous mice to MMTV-Wnt-1 transgenic mice that routinely develop ductal carcinomas in the mammary gland. Female Wnt-1 transgenics heterozygous for Pten developed mammary tumors earlier than Wnt-1 transgenics that were wild type for Pten. In most tumors arising in Pten heterozygotes, the Pten wild-type allele was lost, suggesting that cells lacking Pten function have a growth advantage over cells retaining a wild type allele. Tumors with LOH contained high levels of activated AKT/PKB, a downstream target of the PTEN/PI3K pathway. CONCLUSIONS: An animal model has been developed in which the absence of Pten collaborates with Wnt-1 to induce ductal carcinoma in the mammary gland. This animal model may be useful for testing therapies specific for tumors deregulated in the PTEN/PI3K/AKT pathway.

Motion direction signals in the primary visual cortex of cat and monkey.

When an image feature moves with sufficient speed it should become smeared across space, due to temporal integration in the visual system, effectively creating a spatial motion pattern that is oriented in the direction of the motion. Recent psychophysical evidence shows that such "motion streak signals" exist in the human visual system. In this study, we report neurophysiological evidence that these motion streak signals also exist in the primary visual cortex of cat and monkey. Single neuron responses were recorded for two kinds of moving stimuli: single spots presented at different velocities and drifting plaid patterns presented at different spatial and temporal frequencies. Measurements were made for motion perpendicular to the spatial orientation of the receptive field ("perpendicular motion") and for motion parallel to the spatial orientation of the receptive field ("parallel motion"). For moving spot stimuli, as the speed increases, the ratio of the responses to parallel versus perpendicular motion increases, and above some critical speed, the response to parallel motion exceeds the response to perpendicular motion. For moving plaid patterns, the average temporal tuning function is approximately the same for both parallel motion and perpendicular motion; in contrast, the spatial tuning function is quite different for parallel motion and perpendicular motion (band pass for the former and low pass for the latter). In general, the responses to spots and plaids are consistent with the conventional model of cortical neurons with one rather surprising exception: Many cortical neurons appear to be direction selective for parallel motion. We propose a simple explanation for "parallel motion direction selectivity" and discuss its implications for the motion streak hypothesis. Taken as a whole, we find that the measured response properties of cortical neurons to moving spot and plaid patterns agree with the recent psychophysics and support the hypothesis that motion streak signals are present in V1.