The histone demethylase enzymes KDM3A and KDM4B co-operatively regulate chromatin transactions of the estrogen receptor in breast cancer

Many estrogen receptor (ER)-positive breast cancers develop resistance to endocrine therapy but retain canonical receptor signalling in the presence of selective ER antagonists. Numerous co-regulatory proteins, including enzymes that modulate the chromatin environment, control the transcriptional activity of the ER. Targeting ER co-regulators has therefore been proposed as a novel therapeutic approach. By assessing DNA-binding dynamics in ER-positive breast cancer cells, we have identified that the histone H3 lysine 9 demethylase enzymes, KDM3A and KDM4B, co-operate to regulate ER activity via an auto-regulatory loop that facilitates the recruitment of each co-activating enzyme to chromatin. We also provide evidence that suggests that KDM3A primes chromatin for deposition of the ER pioneer factor FOXA1 and recruitment of the ER-transcriptional complex, all prior to ER recruitment, therefore establishing an important mechanism of chromatin regulation involving histone demethylases and pioneer factors, which controls ER functionality. Importantly, we show via global gene-expression analysis that a KDM3A/KDM4B/FOXA1 co-regulated gene signature is enriched for pro-proliferative and ER-target gene sets, suggesting that abrogation of this network could be an efficacious therapeutic strategy. Finally, we show that depletion of both KDM3A and KDM4B has a greater inhibitory effect on ER activity and cell growth than knockdown of each individual enzyme, suggesting that targeting both enzymes represents a potentially efficacious therapeutic option for ER-driven breast cancer

COMPARING FUNCTIONAL DATA ANALYSIS AND HYSTERESIS LOOPS WHEN TESTING TREATMENTS FOR REDUCING HEAT STRESS IN DAIRY COWS

Various techniques are commonly used to reduce heat stress, including sprayers and misters, shading, and changes in feed. Oftentimes studies are performed where researchers do not control the times when animals use shading or other means available to reduce heat stress, making it hard to test differences between treatments. Two methods are used on data from a study where Holstein cows were given free access to weight activated “cow showers.” Functional data analysis can be used to model body temperature as a function of time and environmental variables such as the Heat Load Index. Differences between treatment groups can be tested using a Functional Bayesian MCMC model. Alternatively hysteresis loops, such as the ellipse, formed by a plot of air temperature or the Heat Load Index against body temperature over the course of a day can be estimated and their parameters used to test differences between cows with access to showers and cows without. Results from an R package hysteresis, which can estimate these loops and their parameters are illustrated. Functional data analysis allows for looser assumptions regarding the body temperature curve and the ability to look for differences between groups at specific time points, while hysteresis loops give the ability to look at heat stress over the course of a day holistically in terms of parameters such as amplitude, lag, internal heat load and central values

COMPARISON OF SAS PROC NLIN AND NLMIXED FOR PARAMETER ESTIMATION IN PET MODEL

Cattle body temperatures were measured under semi-controlled hot cyclic chamber temperatures. The four-parameter nonlinear PET model, is used to estimate body temperature in cattle challenged by heat stress. For each steer, the parameters can be estimated by the Nlin procedure and the sensitivity of each animal can be studied. It is also desirable to generalize the results by using the Nlmixed procedure to combine both the fixed and random effects. When comparing the results from the two procedures, we found heterogeneity among animals and/or days caused convergence problems for proc Nlmixed. Simulation studies were used to study how deviations from homogeneity effected the accuracy of parameter estimates, coverage of confidence intervals, and measures of nonlinear behavior when using the PET model to describe the dynamics of heat stress in cattle

Effects of selenium and vitamin E on performance, physiological response, and selenium balance in heat-stressed sheep

Forty-two 7-mo-old Australian Merino wethers were used in a 50-d trial to investigate the effects of Se and vitamin E on the performance and physiological responses of heat-stressed sheep. Sheep were exposed to thermoneutral conditions (maximum = 24°C and minimum = 20°C) for 28 d followed by heat (maximum = 38°C and minimum = 28°C) for 22 d. Hot conditions were imposed between 0700 and 1800 h. Sheep were randomly allocated to diets containing 0.8 mg/kg Se (Sel-Plex), 150 mg/kg vitamin E, or 0.8 mg/kg Se and 150 mg/kg vitamin E for either the duration of the study (50 d) or from d 1 of the hot period until the end of the study. A control group that received no supplemental Se and vitamin E for the duration of the study was included. Feed intake was measured daily and sheep were weighed weekly. Blood samples were collected from all sheep before feeding on d 1, 21, and 49 for measurement of biochemical and enzymatic variables. The concentration of Se was determined in offered and refused feed, feces, urine, water, plasma, liver, and kidneys. Exposure to heat reduced (P < 0.05) DMI by 11.9%, ADG by 198 g, serum concentration of urea nitrogen and Se by 17.8%, and plasma total antioxidant status by 26.4%. During hot conditions, sheep receiving Se and vitamin E supplements for 50 d had reduced (P < 0.05) BW loss and elevated G:F compared to control sheep. Serum Se concentration and the plasma total antioxidant status were greatest in sheep receiving Se and vitamin E supplements for 50 d (P < 0.05). These results indicate that dietary supplementation with Se and vitamin E reduces the adverse effects of a high heat load. Additional studies are warranted to elucidate the mechanisms responsible for these effects

Spatial distribution of shore-based fishers in the greater Perth Metropolitan area over summer 2010/2011

Aerial surveys are a useful technique for estimating numbers of recreational fishers. Following the success of a pilot study to examine the spatial patterns of recreational shore-based fishing activity in the Perth Metropolitan area from April – June 2010, another survey was conducted from December 2010 – February 2011 between Lancelin – Bunbury. More than 7,000 shorebased fishers were counted during 36 aerial flights, with significantly more recorded on morning and afternoon flights, when compared to those conducted at midday. Flights on weekends/public holidays also recorded more fishers than weekdays. The spatial distribution of fishers was variable along the coast, with sandy beaches at the northern and southern sections of the study area having high counts of fishers, along with groynes in the central section. Comparisons with the previous (autumn) study revealed that the northern beaches generally had higher numbers of shore-based fishers in summer, while those in the south had more during autumn. Data from this study assists with understanding the patterns of shore-based recreational fishing occurring along the greater Perth Metropolitan coast. Such information can be used to assist in the design of future on-ground surveys, compliance and education activities

Seat Belt Use by Missouri Teens*

In 2006, the Institute of Public Policy conducted 12 focus groups with Missouri teens on behalf of the Missouri Department of Transportation to:1) understand how teenagers make decisions on seatbelt use; and 2) determine what outreach methods could influence teens' decisions regarding seatbelt use and traffic safety. The study found that teens' seatbelt use is set well before they begin driving, that the lack of use of seatbelts among parents is especially influential, and that teens are not well informed about the consequences of accidents when drivers or passengers are not wearing seatbelts. This report summarizes studies of seatbelt use in Missouri as compared to other states and describes teens' attitudes about seatbelt usage. Finally, it recommends that the Department continue its teen- focused informational campaign and that the state enact primary enforcement of the seatbelt law.Includes bibliographical reference

CHARACTERIZING THERMAL HYSTERESIS IN BODY TEMPERATURE FOR A HEAT STRESSED STEER

Studies have shown that exposure of animals to a high ambient temperature environment poses serious threats to their health, performance and productivity. Above a certain threshold an animal\u27s body temperature (Tb) appears to be driven by the hot ambient temperature (Ta). For steers challenged by heat stress, the Tb-Ta relationship shows a dramatic increase in Tb per unit change of Ta and the dynamics of the Tb-Ta relationship follow a pattern which depends on whether Ta is increasing or decreasing. A delay becomes noticeable in a steer’s thermo-regulatory response to Ta when Ta is controlled to be sinusoidal in the steer’s heat stress chamber. In other words, Tb lags behind Ta. Consequently when plotted in a Tb-Ta phase diagram, a hysteresis effect appears in the form of a hysteresis loop, indicating the steer is thermally challenged. The hysteresis loop shows a rotated elliptical pattern which depends on the delay (or lag) between Tb and Ta. The angle of rotation of the hysteresis loop indicates the correlation (and lag) between Tb and Ta. The area of the elliptical hysteresis loop can be used to quantify the amount of heat stress during the period of thermal challenge. For example, results of a thermal challenge of 32±7oC applied to a Hereford steer showed, the delay is longer (4 hr lag) and ellipse is larger in an acute stage than in a chronic stage (3 hr lag). A greater delay (or lag) suggests more time is needed to dissipate the heat stress. This result suggests, steers in an acute stage require more energy to dissipate heat than steers in a chronic stage

REMOTE INFLUENCE OF HUMAN PHYSIOLOGY BY A RITUAL HEALING TECHNIQUE

Two experiments tested the hypothesis that remote calming effects of a traditional healing ritual can be objectively measured using indicators of electrodermal activity, heart rate and blood volume. A total of 14 sessions were conducted in the initial study and 16 sessions were conducted in the replication. In both experiments, the authors exchanged roles as experimenter, healer and patient. Healers were instructed to try to calm the remote patient using a set of traditional ritual magic strategies, or to exert no influence (as a control). The patient created a doll in his or her likeness and provided mementos, pictures and an autobiographical sketch. The healer used these materials to form a sympathetic connection with the patient who was located in another building in an isolated toom. During the experiments, there were no other connections between the healer and the patient. Each session consisted of a randomized counterbalanced sequence of five calming and five control epochs of one minute each. No performance feedback was provided to the healer or patient during the session. The combined results of both experiments showed significant effects for changes in blood volume (p = .00002), heart rate (p = .001) and electrodermal activity (p = .013), suggesting that traditional magic healing rituals caused significant relaxation of the vascular system and arousal of electrodermal activity. These rituals appear to be helpful in focusing mental intention in laboratory investigations of direct mental interactions with living systems

The histone demethylase enzyme KDM3A is a key estrogen receptor regulator in breast cancer

Endocrine therapy has successfully been used to treat estrogen receptor (ER)-positive breast cancer, but this invariably fails with cancers becoming refractory to treatment. Emerging evidence has suggested that fluctuations in ER co-regulatory protein expression may facilitate resistance to therapy and be involved in breast cancer progression. To date, a small number of enzymes that control methylation status of histones have been identified as co-regulators of ER signalling. We have identified the histone H3 lysine 9 mono- and di-methyl demethylase enzyme KDM3A as a positive regulator of ER activity. Here, we demonstrate that depletion of KDM3A by RNAi abrogates the recruitment of the ER to cis-regulatory elements within target gene promoters, thereby inhibiting estrogen-induced gene expression changes. Global gene expression analysis of KDM3A-depleted cells identified gene clusters associated with cell growth. Consistent with this, we show that knockdown of KDM3A reduces ER-positive cell proliferation and demonstrate that KDM3A is required for growth in a model of endocrine therapy-resistant disease. Crucially, we show that KDM3A catalytic activity is required for both ER-target gene expression and cell growth, demonstrating that developing compounds which target demethylase enzymatic activity may be efficacious in treating both ER-positive and endocrine therapy-resistant disease