The effect of supplementary selenium source on apparent and true absorption, retention, performance, and selenium status in lactating Holstein cows.

Selenium is an essential trace mineral for dairy cattle and can be provided in the diet in various forms that may differ in bioavailability. The objective of this study was to determine how source of Se affects animal performance, Se status, retention, and apparent and true absorption. Multiparous Holstein cows (n = 24; 597 ± 49 kg body weight) were blocked by days in milk (DIM; 161 ± 18) and randomly assigned to receive 0.3 mg Se/kg of dry matter (100% of NASEM requirements) of either organic Se (ORG; selenized yeast) or inorganic Se (INO; sodium selenite). The Se premix was top-dressed on a common total mixed ration fed daily and mixed into the top 15 cm directly before feeding. Following an 11-wk adaptation period, cows received simultaneous infusions of an intraruminal isotope dose of 77Se in the same chemical form as the premix, and an intravenous dose of 82Se in an inorganic form. Infusions were followed by a 4-d period of blood and rumen fluid sampling, and total collection of feces, urine, and milk. Daily dry matter intake (23 ± 0.6 kg), milk yield (35 ± 1.2 kg), and serum Se (0.11 ± 0.003 µg/g) were not different between treatments during the adaptation period, but milk Se concentrations were greater for ORG compared with INO. Serum 77Se maximum concentration (Cmax) and area under the curve (AUC) were not different between treatments for 72 h following infusion, but rumen fluid 77Se AUC was higher for ORG than INO. Apparent absorption (64 ± 1.4%), and retention (44 ± 1.5%) of the 77Se dose did not differ between treatments. True absorption was calculated using 82Se enrichment in serum and feces and was determined to be 69 ± 1.3% and did not differ between treatments. Fecal excretion of the 77Se dose was not different between treatments (36 ± 1.4%), but ORG had lower urinary excretion and higher milk excretion compared with INO. These results indicate that organic Se resulted in greater Se concentration of milk and lower urinary Se excretion into the environment, but absorption, Se status, and performance of the cow were not affected by Se source at this supplementation level.

Evaluating Osteoarthritis Management Programs: outcome domain recommendations from the OARSI Joint Effort Initiative.

OBJECTIVE: To develop sets of core and optional recommended domains for describing and evaluating Osteoarthritis Management Programs (OAMPs), with a focus on hip and knee Osteoarthritis (OA). DESIGN: We conducted a 3-round modified Delphi survey involving an international group of researchers, health professionals, health administrators and people with OA. In Round 1, participants ranked the importance of 75 outcome and descriptive domains in five categories: patient impacts, implementation outcomes, and characteristics of the OAMP and its participants and clinicians. Domains ranked as "important" or "essential" by ≥80% of participants were retained, and participants could suggest additional domains. In Round 2, participants rated their level of agreement that each domain was essential for evaluating OAMPs: 0 = strongly disagree to 10 = strongly agree. A domain was retained if ≥80% rated it ≥6. In Round 3, participants rated remaining domains using same scale as in Round 2; a domain was recommended as "core" if ≥80% of participants rated it ≥9 and as "optional" if ≥80% rated it ≥7. RESULTS: A total of 178 individuals from 26 countries participated; 85 completed all survey rounds. Only one domain, "ability to participate in daily activities", met criteria for a core domain; 25 domains met criteria for an optional recommendation: 8 Patient Impacts, 5 Implementation Outcomes, 5 Participant Characteristics, 3 OAMP Characteristics and 4 Clinician Characteristics. CONCLUSION: The ability of patients with OA to participate in daily activities should be evaluated in all OAMPs. Teams evaluating OAMPs should consider including domains from the optional recommended set, with representation from all five categories and based on stakeholder priorities in their local context.

Temporal reproducibility of IgG and IgM autoantibodies in serum from healthy women.

Autoantibodies are present in healthy individuals and altered in chronic diseases. We used repeated samples collected from participants in the NYU Women's Health Study to assess autoantibody reproducibility and repertoire stability over a one-year period using the HuProt array. We included two samples collected one year apart from each of 46 healthy women (92 samples). We also included eight blinded replicate samples to assess laboratory reproducibility. A total of 21,211 IgG and IgM autoantibodies were interrogated. Of those, 86% of IgG (n = 18,303) and 34% of IgM (n = 7,242) autoantibodies showed adequate lab reproducibility (coefficient of variation [CV] < 20%). Intraclass correlation coefficients (ICCs) were estimated to assess temporal reproducibility. A high proportion of both IgG and IgM autoantibodies with CV < 20% (76% and 98%, respectively) showed excellent temporal reproducibility (ICC > 0.8). Temporal reproducibility was lower after using quantile normalization suggesting that batch variability was not an important source of error, and that normalization removed some informative biological information. To our knowledge this study is the largest in terms of sample size and autoantibody numbers to assess autoantibody reproducibility in healthy women. The results suggest that for many autoantibodies a single measurement may be used to rank individuals in studies of autoantibodies as etiologic markers of disease.

PIK3CA mutations in plasma circulating tumor DNA predict survival and treatment outcomes in patients with advanced cancers.

BACKGROUND: Oncogenic mutations in PIK3CA are prevalent in diverse cancers and can be targeted with inhibitors of the phosphoinositide-3-kinase/protein kinase B/mammalian target of rapamycin (PI3K/AKT/mTOR) pathway. Analysis of circulating tumor DNA (ctDNA) provides a minimally invasive approach to detect clinically actionable PIK3CA mutations. PATIENTS AND METHODS: We analyzed PIK3CA hotspot mutation frequency by droplet digital PCR (QX 200; BioRad) using 16 ng of unamplified plasma-derived cell-free DNA from 68 patients with advanced solid tumors (breast cancer, n = 41; colorectal cancer, n = 13; other tumor types, n = 14). Results quantified as variant allele frequencies (VAFs) were compared with previous testing of archival tumor tissue and with patient outcomes. RESULTS: Of 68 patients, 58 (85%) had PIK3CA mutations in tumor tissue and 43 (74%) PIK3CA mutations in ctDNA with an overall concordance of 72% (49/68, κ = 0.38). In a subset analysis, which excluded samples from 26 patients known not to have disease progression at the time of sample collection, we found an overall concordance of 91% (38/42; κ = 0.74). PIK3CA-mutated ctDNA VAF of ≤8.5% (5% trimmed mean) showed a longer median survival compared with patients with a higher VAF (15.9 versus 9.4 months; 95% confidence interval 6.7-17.1 months; P = 0.014). Longitudinal analysis of ctDNA in 18 patients with serial plasma collections (range 2-22 time points, median 5) showed that those with a decrease in PIK3CA VAF had a longer time to treatment failure (TTF) compared with patients with an increase or no change (10.7 versus 2.6 months; P = 0.048). CONCLUSIONS: Detection of PIK3CA mutations in ctDNA is concordant with testing of archival tumor tissue. Low quantity of PIK3CA-mutant ctDNA is associated with longer survival and a decrease in PIK3CA-mutant ctDNA on therapy is associated with longer TTF.

The Effect of Dye and Pigment Concentrations on the Diameter of Melt-Electrospun Polylactic Acid Fibers.

Sub-microfibers and nanofibers produce more breathable fabrics than coarse fibers and are therefore widely used in the textiles industry. They are prepared by electrospinning using a polymer solution or melt. Solution electrospinning produces finer fibers but requires toxic solvents. Melt electrospinning is more environmentally friendly, but is also technically challenging due to the low electrical conductivity and high viscosity of the polymer melt. Here we describe the use of colorants as additives to improve the electrical conductivity of polylactic acid (PLA). The addition of colorants increased the viscosity of the melt by >100%, but reduced the electrical resistance by >80% compared to pure PLA (5 GΩ). The lowest electrical resistance of 50 MΩ was achieved using a composite containing 3% (w/w) indigo. However, the thinnest fibers (52.5 µm, 53% thinner than pure PLA fibers) were obtained by adding 1% (w/w) alizarin. Scanning electron microscopy revealed that fibers containing indigo featured polymer aggregates that inhibited electrical conductivity, and thus increased the fiber diameter. With further improvements to avoid aggregation, the proposed melt electrospinning process could complement or even replace industrial solution electrospinning and dyeing.

Precise localization of silicone-based intercranial planar electrodes in magnetic resonance imaging.

Intercranial planar electrodes enable neural recordings with high spatial resolution in diagnosis as well as for treatments. The value of the measurements increases with the precision of localization of the electrodes related to the individual anatomy. In this context, post-implantation MRI provides excellent soft tissue contrast, but the accurate localization of electrodes is impaired by magnetic susceptibility artifacts. We have addressed this problem without adding a substantial burden to the electrode fabrication process. Simple silicone reference structures were strategically placed on the implant surface to visualize the electrodes position in MRI. These reference structures allowed high precision electrode localization independently of electrode imaging artifacts. This implant manufacturing approach could prove extremely useful in combination with existing image processing pipelines.

Effects of encapsulated nitrate on growth performance, nitrate toxicity, and enteric methane emissions in beef steers: Backgrounding phase.

A long-term experiment was conducted to examine the effects of feeding encapsulated nitrate (EN) on growth, enteric methane production, and nitrate (NO) toxicity in beef cattle fed a backgrounding diet. A total of 108 crossbred steers (292 ± 18 kg) were blocked by BW and randomly assigned to 18 pens. The pens (experimental unit; 6 animals per pen) received 3 dietary treatments: Control, a backgrounding diet supplemented with urea; 1.25% EN, control diet supplemented with 1.25% encapsulated calcium ammonium NO (i.e., EN) in dietary DM, which partially replaced urea; or 2.5% EN, control diet supplemented with 2.5% EN (DM basis) fully replacing urea. Additionally, 24 steers were located in 4 pens and randomly assigned to 1 of the above 3 dietary treatments plus a fourth treatment: 2.3% UEN, control diet supplemented with 2.3% unencapsulated calcium ammonium NO (UEN) fully replacing urea. Animals in the additional 4 pens were used for methane measurement in respiratory chambers, and the pens (except UEN) were also part of the performance study (i.e., = 7 pens/treatment). The experiment was conducted for 91 d in a randomized complete block design. During the experiment, DMI was not affected by inclusion of EN in the diet. Feeding EN had no effect on BW, ADG, and G:F ( ≥ 0.57). Methane production (g/d) tended to decrease ( = 0.099) with EN and UEN, but yield (g/kg DMI) did not differ ( = 0.56) among treatments. Inclusion of EN in the diet increased ( ≤ 0.02) sorting of the diets in favor of large and medium particles and against small and fine particles, resulting in considerable increases in NO concentrations of orts without affecting DMI. Plasma NO-N and NO-N concentrations increased ( ≤ 0.05) for EN compared with Control in a dose response manner, but blood methemoglobin levels were below the detection limit. Nitrate concentration in fecal samples slightly increased (from 0.01% to 0.14% DM; < 0.01) with increasing levels of EN in the diet. In conclusion, EN can be used as a feed additive replacing urea in beef cattle during a backgrounding phase in the long term without NO intoxication or any negative effects on growth performance. In addition, the study confirmed that feeding EN tended to decrease enteric methane production in the long term.

Effects of encapsulated nitrate on growth performance, carcass characteristics, nitrate residues in tissues, and enteric methane emissions in beef steers: Finishing phase.

A finishing feedlot study was conducted with beef steers to determine effects of encapsulated nitrate (EN) on growth performance, carcass characteristics, methane production, and nitrate (NO) residues in tissues. The 132 crossbred steers were backgrounded in a feedlot for 91 d and transitioned for 28 days to the high-concentrate diets evaluated in the present study, maintaining the treatment and pen assignments designated at the start of the backgrounding period. The steers were initially assigned to 22 pens (6 animals per pen) in a randomized complete block design with BW (18 pens) and animals designated for methane measurement (4 pens) as blocking factors. Five animals in each pen designated for methane measurement (total of 20 animals) were monitored for methane emissions in respiratory chambers twice during the experiment. Pens received 3 dietary treatments (7 pens each): Control, a finishing diet supplemented with urea; 1.25% EN, control diet supplemented with 1.25% encapsulated NO in dietary DM that partially replaced urea; and 2.5% EN, control diet supplemented with 2.5% EN (DM basis) fully replacing urea. The final pen designated only for methane measurement received a fourth dietary treatment, 2.3% UEN, the control diet supplemented with unencapsulated NO (UEN) fully replacing urea. The cattle weighed 449 ± SD 32 kg at the start of the 150-d finishing period. The 2.5% EN diet decreased ( < 0.01) DMI compared with Control and 1.25% EN diets. Feeding EN tended to increase ( = 0.092) ADG compared with Control, and G:F was improved ( < 0.01) for EN compared with Control. No differences in methane production (g/d) and yield (g/kg DMI) were observed among treatments. Inclusion of EN in the diets increased ( ≤ 0.03) sorting in favor of large and medium particles and against small and fine particles. Plasma NO and NO concentrations were elevated ( < 0.01) with EN in a dose-response manner, but total blood methemoglobin levels for all treatments were low, below the detection limit. Feeding EN increased ( < 0.01) NO concentrations of samples from muscle, fat, liver, and kidney; NO concentrations of these tissues were similar between 1.25% EN and 2.3% UEN. In conclusion, inclusion of 2.5% EN in a finishing diet (DM basis; about 2% NO) did not cause NO toxicity or any health problems in the long term. In comparison with supplemental urea, feeding EN improved feed efficiency despite increases in sorting against dietary EN.

Predicting fecal nutrient concentrations and digestibilities and growth performance in feedlot cattle by near-infrared spectroscopy.

Fecal nutrients and apparent total tract digestibility (ATTD) were predicted using near-infrared spectroscopy (NIRS) of feces collected from the pen floor or the rectum of feedlot cattle in 2 studies, and pen floor samples were assessed for their ability to predict NE, ADG, and G:F. In study 1, 160 crossbred beef steers in 16 pens (4 pens per treatment) were fed dry-rolled barley or wheat (89% of diet DM) processed at 2 levels. Study 2 utilized 160 crossbred beef steers in 20 pens (5 pens per treatment) that were fed dryrolled barley with 4 levels of barley silage (0%, 4%, 8%, and 12% of diet DM). Both studies fed steers to a target weight of 650 kg. Differences in composition of feces collected from the rectum and the pen floor of a subset of steers (3 to 7) were examined. Fecal pats from the pen floor of each pen were collected throughout the feeding period and composited by pen. Except for DM, which was higher ( 0.01) in pen floor than rectal fecal samples, there were minimal differences in fecal constituents between collection methods. In study 1, interactions between grain type and processing index ( ≤ 0.05) were observed, with a reduction in DM, OM, and starch and an increase in NDF and ADL concentrations being associated with more extensively processed wheat than barley. As grain was more extensively processed, ATTD of all nutrients increased ( 0.01). In study 2, fecal ADF and ADL linearly increased ( 0.01) with increasing silage in the diet, whereas fecal DM and N linearly decreased ( 0.01). Digestibility of all nutrients except starch linearly decreased ( 0.01) with increasing silage. Apparent total tract digestibility of GE predicted using NIRS was related to NEg of the diets as estimated by performance for the wheat-fed steers in study 1 ( = 0.58, = 0.03) and those fed increasing silage in study 2 ( = 0.43, < 0.01). Similarly, observed ADG could be predicted using NIRS for steers fed wheat in study 1 ( = 0.48, = 0.05) and silage in study 2 ( = 0.40, < 0.01), but G:F could not. Using NIRS of feces collected from multiple cattle off the feedlot pen floor demonstrated potential for predicting growth performance of finishing cattle. However, grain type and stage of maturity of the cattle impacted the predictability of equations. Increasing the sample size and sampling frequency may be necessary to improve predictions.

Prevalence, symptom patterns and management of episodic diarrhoea in the community: a population-based survey in 11 countries.

BACKGROUND: The extent of episodic diarrhoea in the community is relatively unknown. AIM: To ascertain the prevalence, symptoms and management behaviours associated with self-reported diarrhoea across 11 countries. METHODS: Community screening surveys were conducted using quota sampling of respondents to identify a nationally representative sample of individuals suffering from 'episodic' diarrhoea (occurring once a month or more often). Second-phase in-depth surveys provided data on epidemiology, symptoms, attributed causes and management of episodic diarrhoea. RESULTS: A total of 11 508 phase 1 and 6613 phase 2 surveys were completed. The prevalence of self-reported episodic diarrhoea ranged from 16% to 23% across the 11 countries. The majority of episodic diarrhoea sufferers were female (57%) and were not diagnosed with pre-existing irritable bowel syndrome (IBS); IBS diagnosis ranged from 9% in Mexico to 44% in Italy. Diarrhoea was frequently attributed to anxiety/stress, food-related causes, gastrointestinal 'sensitivity' and menstruation. Accompanying symptoms included 'stomach pain/cramping' (35-62%), 'stomach grumbling' (29-68%) and 'wind' (18-74%). The proportion of episodic sufferers who reported treating their symptoms with remedies or medications ranged between 46% in Belgium and Canada and 90% in Mexico. CONCLUSIONS: A substantial proportion of the population in middle- to high-income countries report episodic diarrhoea in the absence of a pre-existing diagnosis. These symptoms are likely to be associated with substantial social and economic costs, and have implications on how best to configure and guide self-led, pharmacist-led and primary care management.

Optimization of the measurement procedure during multiphoton tomography of human skin in vivo.

BACKGROUND: The in vivo multiphoton tomography has evolved into a useful tool for the non-invasive investigation of morphological and biophysical characteristics of human skin. Until now, changes of skin have been evaluated mainly by clinical and histological techniques. The current study addresses the effects of a changed acquisition time for single scans in a Z-stack on the directly related qualitative and quantitative interpretability of the data. METHODS: A test area of the skin was used for scanning 12 Z-stacks of 10 volunteers aged between 25 and 34 years. The stacks were taken up to a depth of 220 µm at increments of 10 µm at four different times, 1, 3, 7, 13 s, per scan. Subsequently, the second harmonic generation (SHG)-to-autofluorescence aging index of dermis (SAAID) was evaluated at three different measuring depths, i.e. at the maximum of SHG as well as at depths of 60 and 150 µm. RESULTS: The evaluation did not reveal any significant differences in the SAAID behavior between the Z-stacks of each test area scanned at different acquisition times. However, the acquisition time of 1 s/frame increases the measurement stability without influencing the SAAID behavior. The resolution of subcellular structures decreases significantly at scan times ≤3 s, whereas the acquisition time from 7 to 13 s warrants a high image quality. CONCLUSION: The study has shown that there are no significant differences between the scan speeds per scan in a Z-stack and the resulting SAAID. Acquisition times of 7 s are suitable for the morphological evaluation whereas a further extension to 13 s does not result in any benefits. A scan time per image of 1 s is sufficient for the quantitative evaluation of SAAID thus substantially reducing the possible influence of movement artifacts.

Effects of feed consumption rate of beef cattle offered a diet supplemented with nitrate ad libitum or restrictively on potential toxicity of nitrate.

The objective of the study was to investigate the effects of feed consumption rate on potential toxicity, rumen fermentation, and eating behavior when beef heifers were fed a diet supplemented with nitrate (NI). Twelve ruminally cannulated heifers (827 ± 65.5 kg BW) were used in a randomized complete block design. The experiment consisted of 10-d adaptation, 8-d urea-feeding, and 3-d nitrate-feeding periods. All heifers were fed a diet supplemented with urea (UR) during the adaptation and urea-feeding periods, whereas the NI diet (1.09% NO in dietary DM) was fed during the nitrate-feeding period. After adaptation, heifers were randomly assigned to ad libitum or restrictive feeding (about 80% of ad libitum intake) for the urea- and nitrate-feeding periods. Ad libitum DMI decreased (14.1 vs. 15.1 kg/d; < 0.01) when heifers were fed the NI diet compared with the UR diet. The amount of feed consumed increased ( < 0.01) at 0 to 3 h and decreased ( ≤ 0.03) at 3 to 24 h for restrictive vs. ad libitum feeding of both the UR and NI diets. Compared to the UR diet, the NI diet decreased ( < 0.01) feed consumption at 0 to 3 h and increased ( < 0.02) feed consumption at 3 to 24 h (except feed consumption at 9 to 12 h; = 0.90), indicating nitrate feeding changed the consumption pattern (a more even distribution of feed intake over the day). The increased feed consumption from 0 to 3 h after feeding the NI diet restrictively vs. ad libitum numerically decreased ( = 0.11) rumen pH and numerically or significantly increased ( = 0.01 to 0.28) rumen ammonia, NO, and NO; blood methemoglobin; and plasma NO and NO at 3 h. Regression analysis indicated that increased feed consumption (0 to 3 h) exponentially elevated ( < 0.01; = 0.75) blood methemoglobin, and plasma NO + NO among other rumen and blood variables had the greatest correlation (sigmoid response; < 0.01, = 0.47) with feed consumption (0 to 3 h). Particle size distribution of orts was partially altered ( = 0.02 to 0.40) when the NI diet was fed compared with the UR diet. During the nitrate-feeding period, the nitrate content of orts on d 2 and 3 was greater ( = 0.02) than that on d 1. In conclusion, the increased consumption rate of a diet supplemented with nitrate was an important factor influencing risk of nitrate toxicity based on blood methemoglobin and plasma NO. In addition, the pattern of daily feed consumption was altered by nitrate (creating a "nibbling" pattern of eating) in beef heifers.

Using organic acids to control subacute ruminal acidosis and fermentation in feedlot cattle fed a high-grain diet.

The objective of this study was to determine whether supplementing organic acids can prevent incidences of subacute ruminal acidosis (SARA) in beef heifers fed a diet consisting of 8% barley silage and 92% barley grain-based concentrate (DM basis). Ten ruminally cannulated Hereford crossbred heifers (484 ± 25 kg BW) were used in a replicated 5 × 5 Latin square design with 14-d periods including 10 d for dietary adaptation and 4 d for measurements. Dietary treatments included no supplementation (Control), low fumaric acid (61 g/d), high fumaric acid (125 g/d), low malic acid (59 g/d), and high malic acid (134 g/d). Organic acid supplementation had no effect on DMI ( = 0.77). Similarly, no effects were observed on mean ( = 0.74), minimum ( = 0.64), and maximum ( = 0.27) ruminal pH measured continuously for 48 h. Moreover, area under the curve for pH thresholds 6.2 ( = 0.97), 5.8 ( = 0.66), 5.5 ( = 0.55), and 5.2 ( = 0.93) was similar for all treatments. However, malic acid supplementation lowered the amount of time that ruminal pH was <6.2 compared with the Control ( = 0.02) and fumaric acid treatments ( < 0.01). No effects were observed on total VFA concentrations with organic acid supplementation ( = 0.98) compared with the Control, but greater total VFA concentrations were observed with fumaric acid compared with the malic acid treatments ( = 0.02). The population of total culturable bacteria 3 h after feeding was reduced with supplemental malic acid compared with the Control ( = 0.03) and fumaric acid treatments ( = 0.03). However, no effects were observed with organic acid supplementation on lactic acid-utilizing bacteria ( = 0.59). In conclusion, under the conditions of the present study, organic acid supplementation did not have any significant effects on ruminal fermentation parameters compared with the Control and were not effective in preventing SARA in beef cattle fed high-grain diets.

Risk factors for pneumocystis pneumonia in giant cell arteritis: a single-centre cohort study.

OBJECTIVES: Pneumocystis jiroveci pneumonia (PCP) is a life-threatening opportunistic infection. Few PCP cases in giant cell arteritis (GCA) have been described, but it remains unknown, which patients need PCP prophylaxis. METHODS: Sixty-two patients with GCA from a prospective cohort were studied to identify treatment-related predictors of PCP infection. RESULTS: Four PCP infections occurred, all in patients treated with methotrexate in addition to prednisone. Moreover, PCP is associated with higher cumulative PDN doses and severe lymphocytopenia (<400/µl). CONCLUSIONS: Our findings support PCP-prophylaxis in GCA patients who are treated with methotrexate and PDN, and need high prednisone doses to achieve remission, or develop severe lymphocytopenia.

Effects of encapsulated nitrate on enteric methane production and nitrogen and energy utilization in beef heifers.

UNLABELLED: The objective of this study was to investigate effects of encapsulated nitrate (EN) on enteric methane emission and N and energy utilization in beef heifers. Eight ruminally-cannulated beef heifers (451 ± 21 kg BW) were used in a replicated 4 × 4 Latin square design. Four experimental diets were prepared and fed once daily for ad libitum intake: control, 1%, 2%, and 3% EN (0.15, 0.9, 1.5, and 2.5% NO3(-) in dietary DM, respectively). The control diet (55% forage and 45% concentrate) included encapsulated urea, which was gradually replaced with EN for the EN diets (iso-nitrogenous; 12.5% CP). In each period, EN was increased stepwise by 1% every 4 d during adaptation. A 7-d washout period (control diet offered to all heifers) was provided between experimental periods. Dry matter intake tended to decrease (10.4 to 10.1 kg/d; linear, P = 0.06) with EN levels. Enteric methane yield was linearly decreased (21.3 to 17.4 g/kg DMI; P < 0.01) by EN, and methane production (g/d) recovered to the level from heifers fed the control diet on the first day when EN was withdrawn from the diet. Apparent total-tract digestibility of DM and OM increased (P = 0.03) or tended to increase (P = 0.06), respectively, with EN levels. Starch digestibility tended to be greater (P = 0.07) for EN vs. CONTROL: The concentrations of rumen ammonia-N and plasma urea-N decreased linearly (P < 0.01) with EN. Total urinary N and urea-N excretion as proportions of N intake were linearly decreased (46.3 to 41.4%, = 0.09 and 37.1 to 29.9%, P = 0.01, respectively) with EN addition. However, NO3(-)-N excretion in urine increased linearly (P < 0.01) with EN levels. Fecal N excretion was not affected (P = 0.47) by EN, although fecal NO-N excretion increased linearly (P < 0.01) with inclusion of EN (0.09 to 0.88% of total N, P < 0.01). Retained N tended to be increased (percentage of N intake; 16.6 to 21.4%, = 0.08) by the EN. Supplementary EN lowered (6.64 to 5.46% of GE intake [GEI], P < 0.01) energy losses by enteric methane mitigation, which increased ME supply (calculated; 56.5 to 58.8% of GEI, P = 0.01) without changes in calculated heat production (P = 0.24). As a result, retained energy tended to increase (P = 0.07) with EN levels. In conclusion, feeding EN to beef heifers lowered enteric methane production in a dose-response manner, which slightly increased energy supply. Total urinary N excretion was lowered for EN due to lower urinary urea-N excretion.

Effects of encapsulated nitrate on eating behavior, rumen fermentation, and blood profile of beef heifers fed restrictively or ad libitum.

A slow-release form of nitrate (encapsulated nitrate [EN]) was investigated for effects on risk of toxicity, feed consumption rates, and feed sorting behavior in 2 experiments. In Exp. 1, 5 beef heifers (806 ± 72 kg BW) were fed once daily at 75% of ad libitum intake. Supplementary EN (85.6% DM; 71.4% NO3(-) on a DM basis) was increased by 1% every 4 d from 0 to 1.0, 2.0, 2.9, 3.9, 4.8, and 5.8% of dietary DM (from 10.3 to 15.6% CP and 0.11 to 4.8% total NO3(-)). During the study, a heifer was removed due to nitrate poisoning with 59% blood methemoglobin (MetHb; % of total hemoglobin) at 2.9% EN and another due to refusal to eat the 2% EN diet. When dietary EN increased from 0 to 5.8%, DMI (8.8 to 7.6 kg/d; P < 0.001) and feed consumption from 0 to 3 h were decreased (70.3 to 48.6% of total feed offered on an as-is basis; P = 0.001) and feed consumption from 12 to 24 h was increased (0.6 to 22.6%; P < 0.001). Blood MetHb at 1% EN was negligible (<1.5% of total hemoglobin). However, MetHb levels were greater (average 9.8 vs. 3.1% and maximum 23.6 vs. 13.6% at 3 h) at 2.0 and 2.9% EN than at 3.9, 4.8, and 5.8% EN. In Exp. 2, 8 beef heifers (451 ± 21 kg BW) were used in a replicated 4 × 4 Latin square design and fed for ad libitum intake, once daily, diets containing 0, 1, 2, and 3% EN (isonitrogenous, 12.7% CP, and 0.15 to 2.5% total NO3(-) in dietary DM). Each period consisted of 14 d for adaption and 14 d for sampling followed by a 7-d washout. During adaptation, EN was provided to heifers in a stepwise manner (an increase by 1% every 4 d). In Exp. 2 with ad libitum feeding, feed consumption rates were not different (41.8% of total feed consumed from 0 to 3 h; P = 0.56) among EN levels, but DMI tended to decrease linearly (0 to 3% EN; 10.4 to 10.1 kg/d; P = 0.06) and feed was sorted (linear, P < 0.05) against concentrates (containing EN) with EN inclusion. Potential toxicity of nitrate based on blood MetHb was not observed (all samples <2.0% of total hemoglobin). In both studies, ruminal pH increased numerically (Exp. 1) and significantly (Exp. 2; linear, P = 0.04) with EN. In conclusion, studies indicated that the organoleptic properties of diets containing >3% EN may have caused lower feed intake, reduced feed consumption rates, and feed sorting. In addition, feed consumption rate, which was altered by restrictive or ad libitum feeding, was an important factor affecting the potential toxicity of nitrate (e.g., MetHb). Ad libitum feeding minimized risks of nitrate toxicity.

Inhibition of the phosphoinositide 3-kinase pathway for the treatment of patients with metastatic metaplastic breast cancer.

BACKGROUND: Mesenchymal/metaplastic breast cancers (MpBCs) are often triple-negative (TNBC), and chemo-refractory, and can harbor phosphoinositide 3-kinase (PI3kinase) alterations; thus, therapy with mTor inhibitors may demonstrate activity. PATIENTS AND METHODS: Patients with mesenchymal/MpBC treated with temsirolimus-based regimens were evaluated. Mutational analyses [polymerase chain reaction (PCR)-based DNA sequencing method, mass spectrometric detection (Sequenom MassARRAY), or next-generation sequencing] as well as loss of phosphatase and tensin homolog (PTEN) (immunohistochemistry) were performed (archived tissue when available). RESULTS: Twenty-three patients (one of whom was on two separate trials) were treated using temsirolimus-containing regimens: temsirolimus alone (n = 1 patient) or combined with the following: liposomal doxorubicin and bevacizumab (DAT, n = 18); liposomal doxorubicin (DT, n = 1); paclitaxel and bevacizumab (TAT, n = 2); paclitaxel (TT, n = 1); carboplatin and bevacizumab (CAT, n = 1). Response rate [complete response (CR) + partial response (PR)] was 25% across all regimens; 32% in the anthracycline-based regimens [DAT and DT (CR = 2, PR = 4; N = 19)]. An additional two patients achieved stable disease (SD) ≥6 months [total SD ≥6 months/CR/PR = 8 (33%)]. Molecular aberrations in the PI3K pathway were common: PIK3CA mutation = 6/15 (40%), PTEN mutation = 3/11 (27%), and PTEN loss = 2/11 (18%). A point mutation in the NF2 gene (K159fs*16; NF2 alterations can activate mTor) was found in one patient who attained CR (3+ years). Of the eight patients who achieved SD ≥6 months/CR/PR, all 4 patients with available tissue had a molecular aberration that activate the PIK3CA/Akt/mTOR axis: PIK3CA mutation = 2; PTEN loss = 1; NF2 aberration = 1. CONCLUSIONS: DAT has activity in MpBCs including complete CRs. Molecular aberrations that can activate the PI3 K/Akt/mTOR axis are common in MpBC.

Molecular and phenotypic abnormalities in individuals with germline heterozygous PTEN mutations and autism.

PTEN is a tumor suppressor associated with an inherited cancer syndrome and an important regulator of ongoing neural connectivity and plasticity. The present study examined molecular and phenotypic characteristics of individuals with germline heterozygous PTEN mutations and autism spectrum disorder (ASD) (PTEN-ASD), with the aim of identifying pathophysiologic markers that specifically associate with PTEN-ASD and that may serve as targets for future treatment trials. PTEN-ASD patients (n=17) were compared with idiopathic (non-PTEN) ASD patients with (macro-ASD, n=16) and without macrocephaly (normo-ASD, n=38) and healthy controls (n=14). Group differences were evaluated for PTEN pathway protein expression levels, global and regional structural brain volumes and cortical thickness measures, neurocognition and adaptive behavior. RNA expression patterns and brain characteristics of a murine model of Pten mislocalization were used to further evaluate abnormalities observed in human PTEN-ASD patients. PTEN-ASD had a high proportion of missense mutations and showed reduced PTEN protein levels. Compared with the other groups, prominent white-matter and cognitive abnormalities were specifically associated with PTEN-ASD patients, with strong reductions in processing speed and working memory. White-matter abnormalities mediated the relationship between PTEN protein reductions and reduced cognitive ability. The Pten(m3m4) murine model had differential expression of genes related to myelination and increased corpus callosum. Processing speed and working memory deficits and white-matter abnormalities may serve as useful features that signal clinicians that PTEN is etiologic and prompting referral to genetic professionals for gene testing, genetic counseling and cancer risk management; and could reveal treatment targets in trials of treatments for PTEN-ASD.

Steroid hormone measurements from different types of assays in relation to body mass index and breast cancer risk in postmenopausal women: Reanalysis of eighteen prospective studies.

Epidemiological studies have examined breast cancer risk in relation to sex hormone concentrations measured by different methods: "extraction" immunoassays (with prior purification by organic solvent extraction, with or without column chromatography), "direct" immunoassays (no prior extraction or column chromatography), and more recently with mass spectrometry-based assays. We describe the associations of estradiol, estrone and testosterone with both body mass index and breast cancer risk in postmenopausal women according to assay method, using data from a collaborative pooled analysis of 18 prospective studies. In general, hormone concentrations were highest in studies that used direct assays and lowest in studies that used mass spectrometry-based assays. Estradiol and estrone were strongly positively associated with body mass index, regardless of the assay method; testosterone was positively associated with body mass index for direct assays, but less clearly for extraction assays, and there were few data for mass spectrometry assays. The correlations of estradiol with body mass index, estrone and testosterone were lower for direct assays than for extraction and mass spectrometry assays, suggesting that the estimates from the direct assays were less precise. For breast cancer risk, all three hormones were strongly positively associated with risk regardless of assay method (except for testosterone by mass spectrometry where there were few data), with no statistically significant differences in the trends, but differences may emerge as new data accumulate. Future epidemiological and clinical research studies should continue to use the most accurate assays that are feasible within the design characteristics of each study.

Prevalence of vascular disease in systemic lupus erythematosus compared with type-1 diabetes mellitus: a cross-sectional study of two cohorts.

OBJECTIVES: Systemic lupus erythematosus (SLE) is associated with considerable cardiovascular morbidity that has not yet been directly compared with other diseases with known cardiovascular risk. METHODS: Two hundred and forty-one patients of the multicentre Swiss SLE cohort study (SSCS) were cross-sectionally assessed for coronary heart disease (CHD), cerebrovascular disease (CVD) and peripheral artery disease (PAD). SLE patients were compared with a cohort of 193 patients with type-1 diabetes mellitus being followed at the University Hospital Basel. A subgroup analysis of 50 age- and sex-matched patients from the University Hospital Basel was performed. RESULTS: Of patients within the SSCS 13.3% had one or more vascular events: 8.3% CHD, 5% CVD and 1.2% PAD. In type-1 diabetes mellitus patients, 15% had vascular events: 9.3% CHD, 3.1% CVD and 5.6% PAD. In the matched subgroup, 26% of SLE patients had vascular events (14% CHD) compared with 12% in type-1 DM patients (2% CHD). Cardiovascular risk factors were similar in both groups. Vascular events in SLE patients were associated with age, longer disease duration, dyslipidaemia, and hypertension. CONCLUSION: Cardiovascular morbidity in SLE is at least as frequent as in age- and sex-matched type-1 diabetes mellitus patients. Therefore, aggressive screening and management of cardiovascular risk factors should be performed.