Does CDKN2A loss predict palbociclib benefit?

Palbociclib, an oral small-molecule inhibitor of cyclin-dependent kinases 4 and 6, was recently approved by the U.S. Food and Drug Administration in combination with letrozole for postmenopausal women with advanced hormone receptor-positive, her2-negative breast cancer. Patients with loss of CDKN2A (p16), an inherent negative regulator of cyclin-dependent kinases 4 and 6, were not separately studied because of the significant response of the patients selected based only on receptor status. Here, we report a patient with metastatic estrogen receptor- positive, her2-negative breast cancer with CDKN2A loss who experienced a clinical response to palbociclib.

Effects of feeding ground redberry juniper (Juniperus pinchotii) to gestating ewes on pre- and postpartum performance, serum metabolites and hormones, milk fatty acid composition, and progeny preweaning performance.

The objective of this research was to evaluate effects of replacing sorghum × Sudangrass hay with ground juniper in gestating ewe supplements on pre- and postpartum growth performance, serum metabolites and hormonal concentrations, milk fatty acid composition, and progeny preweaning performance. In a completely randomized design, commercial Rambouillet ewes (age = 3 to 5 yr; initial BW = 65.2 ± 1.6 kg) on a base diet of long-stem sorghum × Sudangrass hay were assigned to 1 of 4 dietary supplements in which ground juniper replaced 0% (CNTL), 33% (18JUN), 66% (36JUN), or 100% (54JUN) of the ground sorghum × Sudangrass hay in a pelleted supplement with ground juniper from d 38 ± 4 of gestation to 2 d postpartum. Treatment DM diet intake overall (g/kg BW) in ewes receiving no juniper was similar ( ≥ 0.38) to that of those receiving increasing concentrations of juniper. Changes in ewe BW and BCS were similar ( ≥ 0.24) in ewes throughout gestation. All serum metabolites and hormones were within normal clinical ranges; however, serum IGF-1 decreased linearly ( = 0.003), alanine increased (linear; = 0.003), and serum Na decreased (linear; = 0.049) as the percentage of juniper increased in the diet. Ewe milk fatty acid composition was similar ( > 0.05) for the majority of fatty acids across treatment groups, with the exception of arachidonic acid (C20:4n6) being greater ( < 0.02) in 54JUN vs. CNTL ewe milk. Lamb birth weights were similar ( = 0.13), whereas lamb ADG tended to differ (quadratic; = 0.06) from d 0 to 14, with 18JUN being the least. At weaning, BW tended ( = 0.09) to linearly decrease in lambs born to ewes consuming greater concentrations of juniper but were not different ( = 0.26) between CNTL and 18JUN, 36JUN, and 54JUN. Results indicated that feeding increasing levels of ground juniper in supplements did not negatively alter ewe performance or serum metabolites and hormones measured pre- and postpartum. Lamb birth weight and preweaning performance appeared unaffected by maternal consumption of ground juniper containing supplements. Results also provide novel information regarding the effects of plant secondary compound consumption throughout pregnancy on ewe and progeny performance and health.

Dodecandrin, a new ribosome-inhibiting protein from Phytolacca dodecandra.

Dodecandrin, a newly discovered ribosome-inhibiting protein, has been isolated and purified from the leaves of the African endod plant, Phytolacca dodecandra. Dodecandrin has a molecular weight of approx. 29 000. It cross-reacts with antiserum prepared against pokeweed antiviral protein from Phytolacca americana and exhibits similar requirements for antiribosomal activity. It is more basic than pokeweed antiviral protein, and comparison of the first 30 amino-terminal residues of the two proteins reveals 83% homology. This level of homology is greater than that between pokeweed antiviral protein and pokeweed antiviral protein S, another antiviral protein found in P. americana. Such conservatism in sequence, coupled with the high efficiency of the proteins in deactivating ribosomes and with their abundance in plant tissue, suggests that they serve an important function in the life of the plant, probably as a defense against infection.

Supervised pharmacy student-led medication review in primary care for patients with type 2 diabetes: a randomised controlled pilot study.

OBJECTIVE: To pilot and feasibility-test supervised final year undergraduate pharmacy student-led medication reviews for patients with diabetes to enable definitive trial design. METHOD: Third year pharmacy students were recruited from one UK School of Pharmacy and trained to review patient's medical records and provide face-to-face consultations under supervision while situated within the patient's medical practice. Patients with type 2 diabetes were recruited by postal invitation letter from their medical practice and randomised via automated system to intervention or usual care. Diabetes-related clinical data, quality of life, patient reported beliefs, adherence and satisfaction with medicines information were collected with validated tools at baseline and 6 months postintervention. The process for collecting resource utilisation data was tested. Stakeholder meetings were held before and after intervention to develop study design and learn from its implementation. Recruitment and attrition rates were determined plus the quality of the outcome data. Power calculations for a definitive trial were performed on the different outcome measures to identify the most appropriate primary outcome measure. RESULTS: 792 patients were identified as eligible from five medical practices. 133 (16.8%) were recruited and randomised to control (n=66) or usual care (n=67). 32 students provided the complete intervention to 58 patients. Initial data analysis showed potential for impact in the right direction for some outcomes measured including glycated haemoglobin, quality of life and patient satisfaction with information about medicines. The intervention was found to be feasible and acceptable to patients. The pilot and feasibility study enabled the design of a future full randomised controlled trial. CONCLUSIONS: Student and patient recruitment are possible. The intervention was well received and demonstrated some potential benefits. While the intervention was relatively inexpensive and provided an experiential learning opportunity for pharmacy students, its cost-effectiveness remains to be determined. TRIAL REGISTRATION NUMBER: ISRCTN26445805; Results.

Effects of Juniperus species and stage of maturity on nutritional, in vitro digestibility, and plant secondary compound characteristics.

Rising feed costs and recurring feed shortages necessitate the investigation into alternative and underutilized feed resources. Nutritional characteristics of species are either unknown or limited to leaves and ground material from small stems. Therefore, the objective was to quantify nutritional characteristics, 48-h true IVDMD (tIVDMD), microbial gas production, and secondary compound characteristics of entire woody plant material of 4 species-, , , and -at immature and mature stages of growth. Immature plants had greater CP concentrations and lower NDF concentrations ( < 0.001) than mature plants regardless of species. Mature plants also had greater ( < 0.001) concentrations of ADF compared with immature plants with the exception of . In general, immature , , and had greater ( < 0.02) tIVDMD and total 48-h and asymptotic gas production than mature plants. Immature and plants were more digested (tIVDMD; < 0.001) than immature and , but tIVDMD did not differ in mature plant material across species. Condensed tannins (CT) were greater ( < 0.001) in immature and than mature plants; differences in CT concentrations among immature species were also detected ( < 0.04). Volatile oil yields were similar across maturity and species with 1 exception: immature yielded more ( < 0.02) volatile oil than mature material. Volatile oil composition across species varied and contained a range of 65 to 70 terpene compounds. The dominant terpenes across species were generally greater ( < 0.05) in immature vs. mature plant material with the exception of . Labdane acids were negligible in , , and and greater in ( < 0.001). Ground material from mature juniper species, although inferior in nutritional quality compared with immature plants, is comparable to traditional low-quality roughage ingredients. Given that has been successfully fed in lamb feedlot diets, the similarities of , and suggest that all three species have potential to be effective roughage ingredients.

The effects of plant polysaccharides and buffer additives on PCR.

A survey of the inhibitory effects of various plant polysaccharides on PCR amplification of a 974-bp section of rbcL in spinach revealed that most of the polysaccharides tested (arabinogalactan, carrageenan, dextran, gum guar, gum karaya, gum locust bean, inulin, mannan, pectin, starch and xylan) were not inhibitory. In contrast, two of the acidic polysaccharides (dextran sulfate and gum ghatti) were inhibitory. The addition of 0.5% Tween 20 reversed the inhibitory effects of gum ghatti (polysaccharide:DNA ratio of 500:1). The inhibitory effect of dextran sulfate (50:1) could be reversed by the addition of Tween 20 (0.25% or 0.5%), DMSO (5%) or polyethylene glycol 400 (5%), but none of these three additives were effective at a 100:1 ratio of dextran sulfate/DNA.

A simple technique for removing plant polysaccharide contaminants from DNA.

A survey of the inhibitory effects of various plant polysaccharides on DNA restrictions (HindIII and EcoRI) revealed that neutral polysaccharides (arabino-galactan, dextran, gum guar, gum locust bean, beta-glucan, inulin, laminaran, mannan and starch) were not very inhibitory. In contrast, acidic polysaccharides (carrageenan, dextran sulfate, gum ghatti, gum karaya, pectin and xylan) were very inhibitory, even at low concentrations. The Elutip-d (RPC-5 type resin) was evaluated for removal of the inhibitory polysaccharides. Used alone or in combination with a phenol/chloroform wash, it proved effective in removing the polysaccharide so that HindIII digestion was possible, except in the cases of carrageenan and dextran sulfate. In addition, the genomic DNA extracts from live oak (Quercus virginiana) and magnolia (Magnolia grandiflora) were sufficiently purified so that the DNAs could be restricted with both EcoRI and HindIII.

Interim and archival preservation of plant specimens in alcohols for DNA studies.

Three plant species, spinach, juniper and broccoli, were stored in different alcohol solutions, and the DNAs were examined to determine changes in the quality and quantity of DNA over time. The smallest changes in the genomic DNA were found in the samples stored in the lower mol wt alcohols (100% methanol and 100% and 95% ethanol) and when proteinase (Pronase E) was used in the extraction procedure. After 11 months of storage in ethanol, excellent yields of high mol wt DNA were obtained, but only when the procedure utilized Pronase E. The use of proteinase was found to be essential to obtain DNA from preserved specimens of spinach and broccoli. This appears to explain the previous reports of failures to obtain DNA from alcohol-preserved specimens. Vacuum infiltration of ethanol resulted in better DNA preservation than passive infiltration of ethanol.

Effect of hyperoxia on substrate utilization during intense submaximal exercise.

Six trained males [mean maximal O2 uptake (VO2max) = 66 ml X kg-1 X min-1] performed 30 min of cycling (mean = 76.8% VO2max) during normoxia (21.35 +/- 0.16% O2) and hyperoxia (61.34 +/- 1.0% O2). Values for VO2, CO2 output (VCO2), minute ventilation (VE), respiratory exchange ratio (RER), venous lactate, glycerol, free fatty acids, glucose, and alanine were obtained before, during, and after the exercise bout to investigate the possibility that a substrate shift is responsible for the previously observed enhanced performance and decreased RER during exercise with hyperoxia. VO2, free fatty acids, glucose, and alanine values were not significantly different in hyperoxia compared with normoxia. VCO2, RER, VE, and glycerol and lactate levels were all lower during hyperoxia. These results are interpreted to support the possibility of a substrate shift during hyperoxia.

Dynamic knee extension as model for study of isolated exercising muscle in humans.

In an attempt to approach a system of isolated exercising muscle in humans, a model has been developed that enables the study of muscle activity and metabolism over the quadriceps femoris (QF) muscles while the rest of the body remains relaxed. The simplest version includes the subject sitting on a table with a rod connecting the ankle and the pedal arm of a bicycle ergometer placed behind the subject. Exercise is performed by knee extension from a knee angle of 90 to approximately 170 degrees while flywheel momentum repositions the relaxed leg during flexion. Experiments where electromyographic recordings have been taken from biceps femoris, gastrocnemius, tibialis anterior, and other muscles in addition to QF indicate that only the QF is active and that there is an equal activation of the lateral, medial, and rectus femoris heads relative to maximum. Furthermore, virtually identical pulmonary O2 uptake (Vo2) during and without application of a pressure cuff below the knee emphasizes the inactivity of the lower leg muscles. The advantages of the model are that all external work can be localized to a single muscle group suitable for taking biopsies and that the blood flow in and sampling from the femoral vein are representative of the active muscles. Thus all measurements can be closely related to changes in the working muscle. Using this model we find that a linear relationship exists between external work and pulmonary Vo2 over the submaximal range and the maximal Vo2 per kilogram of muscle may be as much as twice as high as previously estimated.

Effect of different incremental exercise protocols on the determination of lactate and ventilatory thresholds.

The aerobic threshold (AeT) determined from ventilatory curves occurs at the same oxygen uptake and is independent of the rate of increase of power output in an incremental exercise test. On the other hand, it has been proposed that the AeT determined from blood lactate curves and the anaerobic threshold (AnT) determined from ventilatory curves may vary depending on the exercise protocol. Seven healthy subjects performed two incremental exercise tests to evaluate the effect of the rate of power increment on the AeT and the AnT determined from the break points in both ventilatory and venous blood lactate curves. The protocols on the cycle ergometer consisted of increments of 15 W every min (slow) or every 0.25 min (fast). When the results were expressed as the corresponding oxygen uptake, neither the lactate AeT (slow = 2.662 +/- 0.395 1/min; fast = 2.577 +/- 0.392 1/min; P greater than 0.05) nor the ventilatory AeT (slow = 2.737 +/- 0.426 1/min; fast = 2.583 +/- 0.555 1/min; P greater than 0.05) was significantly affected by the protocols. The lactate AnT (slow = 3.675 +/- 0.610 1/min; fast = 3.683 +/- 0.610 1/min; P greater than 0.05) and the ventilatory AnT (slow = 3.635 +/- 0.665 1/min; fast = 3.823 +/- 0.645 1/min; P greater than 0.05) were also not significantly affected by the protocols. The oxygen uptake corresponding to a blood lactate concentration of 2 mM was significantly higher for the fast protocol (slow = 3.235 +/- 0.735 1/min; fast = 4.005 +/- 0.643 1/min; P less than 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)

Geographic variation in leaf essential oils and RAPDs of Juniperus polycarpos K. Koch in central Asia.

Variations in the composition of the leaf essential oils and DNA fingerprints (RAPDs) of Juniperus excelsa, J. polycarpos, J. seravschanica, and J. turcomanica were examined. Juniperus procera was also included in the analyses to aid in determining the specific status of J. polycarpos. Based on these analyses, J. polycarpos is recognized as a distinct species from J. excelsa. The common, multi-seeded juniper of central Asia is J. polycarpos. Juniperus seravschanica and J. turcomanica are treated as part of the J. polycarpos complex but are not recognized as formal taxonomic groups at this time. The Balochistan, Pakistan juniper, usually called J. excelsa var. polycarpos or J. macropoda should be referred to as J. polycarpos in the future.

Family planning needs and behavior of Mexican American women: a study of health care professionals and their clientele.

"A random sample of Mexican American women and a sample of family planning health care professionals, both from two major southwestern cities in the United States, were compared in terms of their reports of birth control methods used, problems in obtaining family planning services, and values involved in making fertility-related decisions, within the Mexican American population.... While there were points of agreement between the two samples, discrepancies were found in reports of problems in obtaining family planning services, fertility-related values, and in the acceptability of female sterilization as a birth control method. It was concluded that family planning professionals in these service areas tend to stereotype Mexican American women, and may not yet realize that the family planning attitudes and behavior of these women are probably changing in significant ways." (SUMMARY IN SPA)

Endurance Exercise: Normal Physiology and Limitations Imposed by Pathological Processes (Part 1).

In brief: Endurance exercise induces significant and rapid changes in many physiological functions. To maintain homeostasis, adaptations in the oxygen transport and delivery chain are necessary. Increases in minute ventilation and diffusion across the alveolar-capillary membrane enhance oxygenation of blood in the lungs. Parallel changes in cardiac output, muscle blood flow, and arteriovenous oxygen difference increase oxygen transport and delivery. Pulmonary and cardiovascular diseases can limit oxygen transport and/or delivery, while muscle diseases can impair oxygen delivery.

Endurance Exercise: Normal Physiology and Limitations Imposed by Pathological Processes (Part 2).

In brief: Endurance exercise causes rapid changes in physiological and metabolic functions, involving not only the body's oxygen transport system (discussed in part 1 of this article, August 1986, page 94) but also the fuel supply and endocrine systems. The hormonal changes increase glucose delivery by stimulating glycogenolysis and gluconeogenesis in the liver and glycogen utilization in muscles, and also increase free fatty acid delivery by stimulating lipolysis in fat cells. Patients with disorders of muscle energy metabolism may have difficulty performing either brief, intense exercise or prolonged exercise, depending on the disorder. Diseases that cause abnormal hormonal and autonomic responses to exercise include autonomic neuropathy, central Cushing's disease, and diabetes mellitus.

Effects of using ground redberry juniper and dried distillers grains with solubles in lamb feedlot diets: growth, blood serum, fecal, and wool characteristics.

Effects of using ground redberry juniper and dried distillers grains with solubles (DDGS) in Rambouillet lamb (n = 45) feedlot diets on growth, blood serum, fecal, and wool characteristics were evaluated. In a randomized design study with 2 feeding periods (Period 1 = 64% concentrate diet, 35 d; Period 2 = 85% concentrate diet, 56 d), lambs were individually fed 5 isonitrogenous diets: a control diet (CNTL) that contained oat hay but not DDGS or juniper or DDGS-based diets in which 0 (0JUN), 33 (33JUN), 66 (66JUN), or 100% (100JUN) of the oat hay was replaced by juniper. During Period 1, lambs fed CNTL had greater (P < 0.05) DMI and ADG and tended to have greater (P < 0.10) G:F than lambs fed 0JUN or lambs fed DDGS-based diets. Lamb DMI, ADG, and G:F quadratically increased (P < 0.008) as juniper increased in the DDGS-based diets. During Period 2, lambs fed CNTL had greater (P < 0.05) DMI than lambs fed 0JUN or lambs fed DDGS-based diets, but ADG was similar (P > 0.41). Compared to 0JUN, lambs fed CNTL had similar (P = 0.12) G:F and tended to have less G:F (P = 0.07) than lambs fed DDGS-based diets. Among lambs fed DDGS-based diets, DMI was similar (P > 0.19), ADG increased linearly (P = 0.03), and G:F tended to decrease quadratically (P = 0.06) as juniper increased in the diet. Serum IGF-1, serum urea N (SUN), and fecal N were greater (P < 0.05) and serum Ca and P and fecal P were similar (P > 0.13) for lambs fed CNTL vs. lambs fed DDGS-based diets (CNTL). Within lambs fed DDGS-based diets, SUN increased quadratically (P = 0.01) and fecal N increased linearly (P = 0.004), which can partially be attributed to increased dietary urea and condensed tannin intake. Most wool characteristics were not affected, but wool growth per kilogram of BW decreased quadratically (P = 0.04) as percentage of juniper increased in the DDGS-based diets. When evaluating the entire 91-d feeding trial, results indicated that replacing all of the ground oat hay with ground juniper leaves and stems in lamb growing and finishing diets is not detrimental to animal performance and that DDGS-based diets can reduce total feedlot costs, as compared to sorghum grain and cottonseed meal-based diets. However, compared to using juniper or oat hay as the sole roughage source, using both during the growing period (Period 1) enhanced growth performance and further reduced total feedlot costs.

Feeding selectivity by mantled howler monkeys (Alouatta palliata) in relation to leaf secondary chemistry in Hymenaea courbaril.

This study is a quantitative examination of primate feeding selectivity in relation to secondary chemistry within a single plant species, Hymenaea courbaril. It provides the first evidence that sesquiterpenes may act as feeding deterrents in mantled howler monkeys. A free-ranging group of mantled howler monkeys at the study site of Sector Santa Rosa, Area de Conservacion Guanacaste, Costa Rica were observed for the 2-month period of H. courbaril leaf flush in 1999. Tree characteristic data and leaf specimens were collected from 22 focal trees. Gas chromatography and mass spectrometry were used to estimate relative percentages of sesquiterpenes in leaf specimens. The monkeys fed only on the youngest leaves and only from particular trees. Whereas leaf stage selectivity was likely governed by tannin content and structural carbohydrates in younger and older leaf stages, respectively, differential tree use may be related to variability in sesquiterpene content. There is evidence that alpha-copaene may have played a role in interindividual tree use, and that cyperene may also be implicated. However, there is no reported evidence of antiherbivore activity for cyperene.

Gene flow versus selection pressure and ancestral differentiation in the composition of species: analysis of population variation of Juniperus ashei Buch. using terpenoid data.

Twenty-four populations of Juniperus ashei were sampled throughout the range of this taxon and the terpenoids of the foliage were analyzed by gas/liquid chromatography. Population differentiation was investigated by analysis of variance and numerical taxonomy. Three south Texas and one Mexican population clustered together with the rest of the taxon appearing fairly uniform. No evidence was found of hybridization or introgression with other taxa. Disjunct populations in Oklahoma and the Ozarks, which have been genetically isolated from the central population for thousands of years, showed no signs of differentiation nor genetic drift. The present pattern of distribution probably dates from the Pleistocene. The south Texas and Mexican populations appear to be the primitive elements of the species. Populational differences have apparently been maintained in adjacent populations in spite of seemingly large gene flow and conversely, chemical uniformity is being maintained in many disjunct populations where there is little or no gene flow.

Total and hindlimb oxygen deficit and "repayment" in hypoxic anesthetized dogs.

Whole-body (WB) and hindlimb [(HL), paw excluded] O2 uptake (VO2) were measured in 26 anesthetized paralyzed dogs while they were ventilated with 9% O2-91% N2 for 15- and 30-min periods and with room air during recovery periods. Ten of the dogs were pretreated with 1 mg/kg propranolol (beta-Blockade). O2 deficit during hypoxia and the excess O2 used during recovery were obtained by assuming that VO2 would have followed the time course described by a line connecting prehypoxic and postrecovery VO2. Amounts of O2 deficit and excess were corrected for changes in O2 stores. O2 excess was seldom as great as O2 deficit in either WB or HL and the two quantities were not obviously related. The rate of HL O2 deficit accumulation decreased with time in hypoxia, whereas WB O2 deficit remained constant. All of HL VO2 was attributed to skeletal muscle so that WB O2 deficit and excess could be partitioned into muscle and nonmuscle portions. The rate of nonmuscle O2 deficit accumulation increased with time, but the nonmuscle portion of O2 excess decreased after the longer hypoxic period. beta-Blockade accentuated but did not change these qualitative relationships. We concluded that neither WB nor HL O2 deficits were fully matched by O2 excess and that regional patterns of O2 deficit accumulation and "repayment" did not necessarily parallel those for WB.

O2 transport during two forms of stagnant hypoxia following acid and base infusions.

Cardiac output and mean arterial pressure were decreased in two groups of 16 anesthetized paralyzed dogs ventilated by pump. Pericardial tamponade was used in one group, and hemorrhagic hypotension was used in the other. After a 30-min control period and 30 min of circulatory shock by either method, 0.3N HCl was infused into half the dogs in each group and 1.0N NaHCO3 into the other half so that pH was separated by 0.3-0.4 units. The slope of the line relating O2 uptake to total O2 transport (blood flow X arterial O2 concentration) was used to evaluate how well the tissues extracted O2 relative to O2 supply. During the initial shock period before infusion, the slope of the line relating O2 uptake of left hindlimb skeletal muscle to total O2 transport in the limb was almost twice as great as that for the whole body. Acid infusion increased the slope of the whole-body line but did not alter that for the hindlimb. Base infusion, on the other hand, decreased the slope of the line for the limb during hemorrhagic shock but had no other effect. We concluded that acid either improved the distribution of a limiting blood supply to nonmuscle organ systems, or increased tissue capillary PO2 and O2 diffusion by decreasing hemoglobin O2 affinity (HOA), or both. The effect of an increased HOA with base infusion was noticeable in hindlimb skeletal muscle only when volume depletion by hemorrhage presumably greatly increased the normally short intercapillary diffusion distance in muscle.