Diversity of amino acid signaling pathways on autophagy regulation: A novel pathway for arginine

Autophagy is the intracellular bulk degradation process to eliminate damaged cellular machinery and to recycle building blocks, and is crucial for cell survival and cell death. Amino acids modulate autophagy in response to nutrient starvation and oxidative stress. We investigated the relevance of reactive oxygen species (ROS) production on the regulation of autophagy using amino acids, both as a mixture and individually, in rat hepatoma H4-II-E cells. Nutrient starvation elevated ROS production and stimulated autophagy. Treatment with complete (CAA), regulatory (RegAA) and non-regulatory (NonRegAA) amino acid mixtures showed significant suppression of ROS production, whereas only CAA and RegAA exhibited significant suppression of autophagy, suggesting a dissociation of the two responses. The effects of individual amino acids were examined. Leucine from RegAA decreased ROS production and suppressed autophagy. However, methionine and proline from RegAA and arginine, cystine and glutamic acid from NonRegAA suppressed autophagy with an opposite increase in ROS production. Other amino acids from the NonRegAA group showed stimulating effects on ROS production without an autophagic response. Arginine’s effect on autophagy suppression was not blocked by rapamycin, indicating an mTOR-independent pathway. Inhibitor studies on arginine-regulated autophagy may indicate the involvement of NO pathway, which is independent from ROS and mTOR pathways.     

Resveratrol derivatives increase cytosolic calcium by inhibiting plasma membrane ATPase and inducing calcium release from the endoplasmic reticulum in prostate cancer cells

Resveratrol (RES) is a putative chemotherapeutic naturally found in grapes, peanuts, and Japanese knotweed. Previous studies demonstrate that RES modulates calcium signaling as part of its chemotherapeutic activity. In this study, we determined the chemotherapeutic activity of three RES esters that have been modified at the 4’ hydroxyl by the addition of pivalate, butyrate, and isobutyrate. All of the RES derivatives disrupted the calcium signaling in prostate cancer cells more than the parent compound, RES. Further, we demonstrate that the RES derivatives may disrupt the calcium homeostasis by activating calcium release from the endoplasmic reticulum and inhibiting plasma membrane Ca²⁺-ATPase. The pivalated and butyrated RES derivatives decreased cell viability significantly more than RES. Because pivalated and butyrated RES are more effective than RES at targeting calcium signaling pathways, pivalated and butyrated RES may serve as more effective chemotherapeutics.     

施用不同氮、磷、钾肥和有机肥对‘红阳’猕猴桃生长及产量的影响

为阐明施肥与猕猴桃产量、品质的关系,优化施肥配方,以猕猴桃品种‘红阳’(Actinidia chinensis‘Hongyang’)为试验材料,采用田间小区栽培方法研究了不同施肥处理对‘红阳’植株生长、叶片光合特性及果实产量的影响。研究结果显示,N、P、K任何一种元素的缺乏均会影响植株生长并降低果实产量;均衡施用N、P、K肥可以提高叶片光合作用,促进植株新梢和主干茎的生长,从而提高果实产量;如果N和K比例过大则会降低叶片气孔导度、胞间CO2浓度和净光合速率,从而减少果实产量;增施有机肥对提高果实产量效果不显著,但可以提高单果重,过量施用有机肥则会显著降低产量。本研究结果表明,均衡施用N、P、K并配施一定的有机肥(N 250 g/株、P2O5250 g/株、K2O 250 g/株、有机肥6 kg/株)能显著促进植株的生长并提高果实产量。氮、磷、钾肥与有机肥配合施用是提高猕猴桃果园养分管理的有效方法。     

A Note on the Analysis of a p × 22 Design

TYAGI has given a method of constructing a balanced factorial experiment of the p × 2² type. A complete analysis of such a design with and without recovery of interblock information, using the eigenyalues and eigenvectors of the C-matrix of the design, is given in this paper. KHATRY & SHAH'S method of producing uniformly better estimators is employed to combine the inter and intra block estimates.     

'Population laboratories' or 'laboratory populations'? Making sense of the Baltimore Longitudinal Study of Aging, 1965-1987

Interest among historians, philosophers and sociologists of science in population-based biomedical research has focused on the randomised controlled trial to the detriment of the longitudinal study, the temporally extended, serial observation of individuals residing in the same community. This is perhaps because the longitudinal study is regarded as having played a secondary role in the debates about the validity of populations-based approaches that helped to establish epidemiology as one of the constitutive disciplines of contemporary biomedicine. Drawing on archival data and publications relating to the Baltimore Longitudinal Study of Aging, we argue however that the historical development of the longitudinal study is richer and more significant than has been appreciated. We argue that this history is shaped by the tension between two sets of epistemic practices, devices and norms. On the one side there were those who emphasised randomisation and sampling to evidence claims about, and justify policies with respect to, the aetiology of disease. On the other side there were those who evoked the technical repertoire of physiological research, especially the notion of the ‘model organism’, to argue for a different integration of the individual in modern society.     

Simple cryoprotection and cell dissociation techniques for application of the comet assay to fresh and frozen rat tissues

The single-cell gel electrophoresis (comet) assay has been widely used for genotoxicity studies in cell cultures, but its use in solid tissues is hindered by problems in isolation of cells and in cryopreservation techniques. Here, we used minced liver tissues from rats to compare a homogenization technique for isolation of nuclei with a collagenase digestion method (300 λunits/g liver at 37°C for 20 λmin) for isolation of intact cells for subsequent comet assay. We found that collagenase digestion was preferred to the homogenization technique in fresh tissues, but neither method prevented the extensive DNA damage caused by cryopreservation ( -85°C for 72 λh). To minimize this damage, minced liver (1.0 λg) and kidney (0.5 λg) tissues were added to 20 λml of pre-cooled 10% glycerol or 10% dimethylsulfoxide (DMSO). We showed that cryoprotection with DMSO ( -85°C for 72 λh and 3 weeks), and to a slightly lesser extent with glycerol (72 λh), followed by collagenase digestion led to satisfactory recovery of liver cells with little or no DNA strand breakage. We then used DMSO as a cryoprotective agent to optimize the amount of collagenase and its incubation time in frozen liver and kidney tissues. We showed that the collagenase digestion at 150 λunits/g liver and 300 λunits/g kidney for 10 λmin produced highest cell numbers and minimal DNA strand breaks. We also validated these procedures by injection (i.p.) of rats with a known renal carcinogen, ferric nitrilotriacetate (Fe/NTA). We showed that Fe/NTA strongly induced DNA strand breaks in both rat liver and kidney, while no DNA strand breakage occurred in these tissues from the control rats. In addition, no significant differences in strand breaks were found between fresh tissues and tissues treated with DMSO during freezing at -85°C for 72 λh. Thus, the cryoprotection and the cell dissociation techniques developed here are satisfactory for preparing both fresh and frozen tissues for comet assay. These simple techniques are expected to expand greatly the usefulness and efficacy of the assay.     

Optimal adaptive randomized designs for clinical trials

Optimal decision-analytic designs are deterministic. Such designsare appropriately criticized in the context of clinical trialsbecause they are subject to assignment bias. On the other hand,balanced randomized designs may assign an excessive number ofpatients to a treatment arm that is performing relatively poorly.We propose a compromise between these two extremes, one thatachieves some of the good characteristics of both. We introducea constrained optimal adaptive design for a fully sequentialrandomized clinical trial with k arms and n patients. An r-designis one for which, at each allocation, each arm has probabilityat least r of being chosen, 0 r 1/k. An optimal design amongall r-designs is called r-optimal. An r₁-design is also an r₂-designif r₁ r₂. A design without constraint is the special case r = 0and a balanced randomized design is the special case r = 1/k.The optimization criterion is to maximize the expected overallutility in a Bayesian decision-analytic approach, where utilityis the sum over the utilities for individual patients over a‘patient horizon’ N. We prove analytically thatthere exists an r-optimal design such that each patient is assignedto a particular one of the arms with probability 1 – (k – 1)r,and to the remaining arms with probability r. We also show thatthe balanced design is asymptotically r-optimal for any givenr, 0 r < 1/k, as N/n  . This implies that everyr-optimal design is asymptotically optimal without constraint.Numerical computations using backward induction for k = 2arms show that, in general, this asymptotic optimality featurefor r-optimal designs can be accomplished with moderate trialsize n if the patient horizon N is large relative to n. We alsoshow that, in a trial with an r-optimal design, r < 1/2,fewer patients are assigned to an inferior arm than when followinga balanced design, even for r-optimal designs having the samestatistical power as a balanced design. We discuss extensionsto various clinical trial settings.     

Physiological energetics of the green abalone, Haliotis fulgens, fed on a balanced diet

A regression of different physiological responses against body size enables populations or stock cultures of various size ranges to be compared. Thus, the aim of the present work was to evaluate the physiological responses from juvenile green abalone (Haliotis fulgens), grouped according to size, in a standard controlled culture condition within their optimal range to create the balanced growth equation within an allometric relation, providing a basic framework for physiological research into H. fulgens. Feed intake, absorption efficiency, respiration and excretion were measured as functions of dry tissue weight (DTW) in juvenile abalone acclimated on a balanced diet. The daily feed intake (I) was related to body mass by the relationship I (mg day⁻¹)=24.25W^0.59. Absorption efficiency averaged 87% and was independent of body size. The equation relating daily respiration rate (R) to body mass was R (ml O₂ day⁻¹)=12.013W^0.704, including night correction. The rate of ammonia excretion (U) was related to dry tissue weight by the equation U (μmol N-NH₄ day⁻¹)=43.57W^0.85. The energetic value of food was 18.8 J mg⁻¹. The low feeding activity (<1% of abalone live weight per day) was compensated for by a relatively high absorption efficiency and a depression of 23% in the metabolic rate during diurnal activity. Gross and net growth efficiencies were constant throughout the size range, with average values of 36.4% and 41.5%, respectively. Values of the O/N ratio, with an average of 31.5, showed a higher use of proteins from the diet as an energy source for the size range studied.     

宋内Ⅰ相O抗原及霍乱毒素B亚基在福氏2a痢疾菌中的表达及其免疫保护效果的观察

本文利用基因重组的方法,将宋内Ｉ相Ｏ抗原基因以及霍乱毒素Ｂ亚单位基因（ｃｔｘ－Ｂ）克隆至带链球菌的ａｓｄ基因的表达载体,然后转化至ａｓｄ－痢疾菌苗株福氏２ａＴ３２。脂多糖银染以及Ｗｅｓｔｅｒｎｂｌｏｔｔｉｎｇ实验证实以上两基因都能在宿主菌中稳定表达。动物（小白鼠）保护实验表明,该重组菌对福氏２ａ、宋内氏痢疾菌的保护效率达１００％,对霍乱弧菌的保护效率也达７０％。该菌具有稳定、无抗生素标记、多价的特点。