Optimum Designs for Estimating the Variance Components in a Simple Variance Component Model (Model II), I

The methods of optimal designing of experiments proposed by WALD (1943) are used for determination of an A_σ2-optimal concrete design for estimation of σ^2′ = (σ, σ) in case of one-way analysis of variance. Starting point of definition of the optimality criterion is a quadratic loss matrix.     

静注人免疫球蛋白细菌内毒素检查方法的建立

目的 :建立用于静脉注射用人免疫球蛋白 (IVIG)的细菌内毒素检查方法。方法 :通过干扰评价实验证明IVIG对细菌内毒素检查法有强烈的抑制作用 ,单纯用简单稀释和调整pH值法无法消除干扰 ,如果用稀释剂 (Ⅰ )对IVIG作 1∶4以上的稀释 ,即可消除样品对该检查法的干扰 ,利用灵敏度为 0 1 2 5EU ml的鲎试剂 ,将样品用稀释剂 (Ⅰ )稀释 4倍 ,按中国药典 ( 2 0 0 0版 )进行细菌内毒素的检查 ,结果与家兔热原质试验进行比较。结论 :IVIG用稀释剂 (Ⅰ )进行适当稀释后 ,可以用细菌内毒素检查法替代家兔热原试验 ,应用于生产过程及半成品的质量控制。     

A Simple Method to Obtain all the Sample-Values and their Probabilities of the Wald-Wolfowitz Test Statistics in the Existence of Ties

In order to carry out non-conservative tests in the general two-sample problem with ties, we want to know all possible sample-values of the used test statistics and their occurrence probabilities as well. But this knowledge can be acquired only after very protracted attempts. In the present paper we depict a simple technique for obtaining that without any exertion in the case of the Wald-Wolfowitz test statistic. With that, we then are able to lead the Wald-Wolfowitz test easily and effortlessly in any manner conservative or non-conservative and in the existence of any number and any length of ties.     

Reliability and Validity of a 20-s Alternative to the Wingate Anaerobic Test in Team Sport Male Athletes

The intent of this study was to evaluate relative and absolute reliability of the 20-s anaerobic test (WAnT₂₀) versus the WAnT₃₀ and to verify how far the various indices of the 30-s Wingate anaerobic test (WAnT₃₀) could be predicted from the WAnT₂₀ data in male athletes. The participants were Exercise Science majors (age: 21.5±1.6 yrs, stature: 0.183±0.08 m, body mass: 81.2±10.9 kg) who participated regularly in team sports. In Phase I, 41 participants performed duplicate WAnT₂₀ and WAnT₃₀ tests to assess reliability. In Phase II, 31 participants performed one trial each of the WAnT₂₀ and WAnT₃₀ to determine the ability of the WAnT₂₀ to predict components of the WAnT₃₀. In Phase III, 31 participants were used to cross-validate the prediction equations developed in Phase II. Respective intra-class correlation coefficients (ICC) for peak power output (PPO) (ICC = 0.98 and 0.95) and mean power output (MPO) (ICC 0.98 and 0.90) did not differ significantly between WAnT₂₀ and WAnT₃₀. ICCs for minimal power output (PO_min) and fatigue index (FI) were poor for both tests (range 0.53 to 0.76). Standard errors of the means (SEM) for PPO and MPO were less than their smallest worthwhile changes (SWC) in both tests; however, PO_min and FI values were “marginal,” with SEM values greater than their respective SWCs for both tests values. Stepwise regression analysis showed that MPO had the highest coefficient of predictability (R = 0.97), with PO_min and FI considerable lower (R = 0.71 and 0.41 respectively). Cross-validation showed insignificant bias with limits of agreement of 0.99±1.04, 6.5±92.7 W, and 1.6±9.8% between measured and predicted MPO, PO_min, and FI, respectively. WAnT₂₀ offers a reliable and valid test of leg anaerobic power in male athletes and could replace the classic WAnT₃₀.     

Simple Fecal Flotation Is a Superior Alternative to Guadruple Kato Katz Smear Examination for the Detection of Hookworm Eggs in Human Stool

Background

Microscopy-based identification of eggs in stool offers simple, reliable and economical options for assessing the prevalence and intensity of hookworm infections, and for monitoring the success of helminth control programs. This study was conducted to evaluate and compare the diagnostic parameters of the Kato-Katz (KK) and simple sodium nitrate flotation technique (SNF) in terms of detection and quantification of hookworm eggs, with PCR as an additional reference test in stool, collected as part of a baseline cross-sectional study in Cambodia.

Methods/Principle Findings

Fecal samples collected from 205 people in Dong village, Rovieng district, Preah Vihear province, Cambodia were subjected to KK, SNF and PCR for the detection (and in case of microscopy-based methods, quantification) of hookworm eggs in stool. The prevalence of hookworm detected using a combination of three techniques (gold standard) was 61.0%. PCR displayed a highest sensitivity for hookworm detection (92.0%) followed by SNF (44.0%) and quadruple KK smears (36.0%) compared to the gold standard. The overall eggs per gram feces from SNF tended to be higher than for quadruple KK and the SNF proved superior for detecting low egg burdens.

Conclusion/Significance

As a reference, PCR demonstrated the higher sensitivity compared to SNF and the quadruple KK method for detection of hookworm in human stool. For microscopic-based quantification, a single SNF proved superior to the quadruple KK for the detection of hookworm eggs in stool, in particular for low egg burdens. In addition, the SNF is cost-effective and easily accessible in resource poor countries.     

Genomic Inverse PCR for Exploration of Ligated Breakpoints (GIPFEL), a New Method to Detect Translocations in Leukemia

Here we present a novel method “Genomic inverse PCR for exploration of ligated breakpoints” (GIPFEL) that allows the sensitive detection of recurrent chromosomal translocations. This technique utilizes limited amounts of DNA as starting material and relies on PCR based quantification of unique DNA sequences that are created by circular ligation of restricted genomic DNA from translocation bearing cells. Because the complete potential breakpoint region is interrogated, a prior knowledge of the individual, specific interchromosomal fusion site is not required. We validated GIPFEL for the five most common gene fusions associated with childhood leukemia (MLL-AF4, MLL-AF9, MLL-ENL, ETV6-RUNX1, and TCF3-PBX1). A workflow of restriction digest, purification, ligation, removal of linear fragments and precipitation enriching for circular DNA was developed. GIPFEL allowed detection of translocation specific signature sequences down to a 10⁻⁴ dilution which is close to the theoretical limit. In a blinded proof-of-principle study utilizing DNA from cell lines and 144 children with B-precursor-ALL associated translocations this method was 100% specific with no false positive results. Sensitivity was 83%, 65%, and 24% for t(4;11), t(9;11) and t(11;19) respectively. Translocation t(12;21) was correctly detected in 64% and t(1;19) in 39% of the cases. In contrast to other methods, the characteristics of GIPFEL make it particularly attractive for prospective studies.     

Development of a Simple Culture Method for the Tissues Contaminated with Microorganisms and Application to Establishment of a Fish Cell line

Cell cultures are good in vitro model systems in place of using living animals. In the present study, we developed a simple culture method in which tissues were pretreated with a low concentration of sodium hypochlorite solution (NaClO) to prevent not only bacteria but also fungi. Scales removed from a goldfish (Carassius auratus) body were treated with 70% ethanol and then with 0.3% of sodium hypochlorite solution, and cultured in vitro in an atmosphere containing 0.5% CO2. The doubling time of the established cells (GAKS) was 24 hr. The GAKS cells contained alkaline phosphatase activity (8.3+/-1.1 nmol/min/mg protein) and secreted 0.32+/-0.07 pg/ml endothelin during a 3 day culture of a full monolayer sheet.     

Measurement of Acetylcholine Turnover Rate in Brain: An Adjunct to a Simple HPLC Method for Choline and Acetylcholine

Abstract: An existing method for measuring acetylcholine (ACh) and choline (Ch) is shown to be useful formeasuring the turnover rate of ACh in mouse brain. Methl-[3H]Ch is injected into mice. They are killed atdifferent times by microwave irradiation and Ch and AChextracted and separated by reverse-phase HPLC. Ch andACh are converted to hydrogen peroxide by a post-column enzyme reaction. Hydrogen peroxide, which isdirectly related to the tissue content of Ch or ACh, isdetermined electrochemically. The fractions that corre-spond to the detector response for Ch and ACh are col-lected for the measurement of radioactivity. In this wayspecific radioactivities of endogenous Ch and ACh areestimated in the same sample. We used the specific ra-dioactivity values determined by this procedure to esti-mate the turnover of ACh for striatum, cerebral cortex, and hippocampus of the mouse.     

The need for studies to evaluate the reproducibility of the T-wave alternans (TWA), and the rationale for a correction index of the TWA

Sudden cardiac death (SCD) due to various cardiomyopathies is currently prevented by the implantation of an automated cardioverter/defibrillator (ICD). ICD impalntation in patients who are not survivors of SCD, or have not suffered potentially lethal ventricular arrhythmias, are based on the presence of cardiomyopathy with a reduced left ventricular ejection fraction. The bulk of patients who are considered suitable for an ICD implantation and receive such devices, do not experience device therapy shocks at follow-up ("false positives"), thus creating a climate of uncertainty among patients and physicians about the soundness of our current eligibility criteria for ICDs. In addition the cost of inappropriate ICDs is staggering, and the undue exposure of "false positive" patients to complications, and hardships is disconcerting. T-wave alternans (TWA) has emerged as a possible "risk detection of SCD" technology, but its reproducibility has not been tested. Peripheral edema (extracardiac) or other cardiac mechanisms, unrelated to the degree of SCD risk, alter the amplitude, and other attributes, of the T-waves. Since TWA may be T-wave amplitude-, or other T-wave attributes-dependent (this is still a speculation), a need may be emerging for its correction by the T-wave amplitude (TWA index); such an index may enhance the reproducibility, and evaluate the true sensitivity, specificity and predictive accuracy of the TWA in detecting future victims of SCD.     

Melatonin desensitizing effects on the in vitro responses to MCH,alpha-MSH,isoproterenol and melatonin in pigment cells of a fish (S. marmoratus), a toad (B. ictericus), a frog (R. pipiens), and a lizard (A. carolinensis), exposed to varying photoperiodic regimens

Melatonin is a weak dose-independent lightening agonist in fish skin, a moderate dose-dependent lightening agonist in toad skin and a potent lightening agent in frog and lizard skins (reversing in a dose-dependent manner the darkening caused by alpha-melanocyte-stimulating hormone). In frog skins, previous exposure to melatonin reduced further lightening actions of the indoleamine, and in toad skins, increasing concentrations of melatonin elicited decreasing lightening responses, suggesting an autodesensitizing action of the hormone. Various concentrations of melatonin diminished the responses to the lightening agonist melanin-concentrating hormone (MCH) in fish skins and to the darkening agonists alpha-MSH in toad, frog and lizard skins and isoproterenol in frog skins. In vitro inhibitory actions of melatonin are mimicked in the absence of the hormone in skin preparations from toads kept in continuous darkness for 48 hr. The lipophylic nature of the indoleamine associated with the results herein described suggests intracellular actions of melatonin on vertebrate pigment cells.     

A Test of the Mating Limitation Hypothesis for Caste Determination in Evylaeus albipes (Hymenoptera: Halictidae), a Primitively Eusocial Halictine Bee

Yanega's (1997) mating limitation hypothesis (MLH) states that if a female mates promptly after emerging, she then becomes a member of the maximally reproductive behavioral caste (i.e., in most cases an overwintering gyne). Females that do not mate early become workers. We tested the MLH in laboratory colonies of a eusocial population of Evylaeus albipes. Of 24 worker brood females (13 from queenright and 11 from orphaned nests), 13 mated on the first day of flight activity and all mated within the first 5 days; there were no significant differences between mating rates of females from the two colony types. All 24 commenced foraging as workers after an average of between 3 and 4 days postmating. We conclude that the MLH does not apply to this species despite the fact that the only known halictine for which this hypothesis has been experimentally tested is the fairly closely related E. marginatus.     

The Input Signal Step Function (ISSF), a Standard Method to Encode Input Signals in SBML Models with Software Support, Applied to Circadian Clock Models

Time-dependent light input is an important feature of computational models of the circadian clock. However, publicly available models encoded in standard representations such as the Systems Biology Markup Language (SBML) either do not encode this input or use different mechanisms to do so, which hinders reproducibility of published results as well as model reuse. The authors describe here a numerically continuous function suitable for use in SBML for models of circadian rhythms forced by periodic light-dark cycles. The Input Signal Step Function (ISSF) is broadly applicable to encoding experimental manipulations, such as drug treatments, temperature changes, or inducible transgene expression, which may be transient, periodic, or mixed. It is highly configurable and is able to reproduce a wide range of waveforms. The authors have implemented this function in SBML and demonstrated its ability to modify the behavior of publicly available models to accurately reproduce published results. The implementation of ISSF allows standard simulation software to reproduce specialized circadian protocols, such as the phase-response curve. To facilitate the reuse of this function in public models, the authors have developed software to configure its behavior without any specialist knowledge of SBML. A community-standard approach to represent the inputs that entrain circadian clock models could particularly facilitate research in chronobiology.     

Object Discrimination Reversal As a Method to Assess Cognitive Impairment in Nonhuman Primate Enhanced Pre‐ and Postnatal Developmental (ePPND) Studies: Statistical Power Analysis

An important aspect of the enhanced pre‐ and postnatal developmental (ePPND) toxicity study in nonhuman primates (NHP) is that it combines in utero and postnatal assessments in a single study. However, it is unclear if NHP ePPND studies are suitable to perform all of the evaluations incorporated into rodent PPND studies. To understand the value of including cognitive assessment in a NHP ePPND toxicity study, we performed a power analysis of object discrimination reversal task data using a modified Wisconsin General Testing Apparatus (ODR‐WGTA) from two NHP ePPND studies. ODR‐WGTA endpoints evaluated were days to learning and to first reversal, and number of reversals. With α = 0.05 and a one‐sided t‐test, a sample of seven provided 80% power to predict a 100% increase in all three of the ODR‐WGTA endpoints; a sample of 25 provided 80% power to predict a 50% increase. Similar power analyses were performed with data from the Cincinnati Water Maze (CWM) and passive avoidance tests from three rat PPND toxicity studies. Groups of 5 and 15 in the CWM and passive avoidance test, respectively, provided 80% power to detect a 100% change. While the power of the CWM is not far superior to the NHP ODR‐WGTA, a clear advantage is the routine use of larger sample size, with a group of 20 rats the CWM provides ～90% power to detect a 50% change. Due to the limitations on the number of animals, the ODR‐WGTA may not be suitable for assessing cognitive impairment in NHP ePPND studies.     

The thermoregulatory and metabolic responses to photoperiod manipulations of the Macedonian mouse (Mus macedonicus), a post-fire invader

1. The Macedonian mouse (Mus macedonicus), a small (15 g) mesic rodent distributed in the Mediterranean ecosystem, is a species which invades post fire habitats in the first stages of habitat recovery. 2. In order to assess the seasonal acclimatization of thermoregulatory and metabolic mechanisms, the response of several physiological variables to photoperiod manipulations under a constant T_a were studied. 3. Our results show that mice acclimated to a short photoperiod increased their resistance to cold, while acclimation to a long photoperiod increased their resistance to high T_as and the effectiveness of their thermoregulatory mechanisms. Body mass increased in mice acclimated to a short photoperiod. We conclude that photoperiod is an important environmental cue for seasonal acclimatization of thermoregulatory and metabolic mechanisms in this species.     

The distribution of macroinvertebrate assemblages in a reach of the River Allier (France), in relation to riverbed characteristics

Investigations on large canalised rivers, for example the Danube, have shown that transported particulate matter, which is typically inorganic, is predominantly deposited in waters near the river’s main channel. This investigation deals with the lower section of the River Havel (NE Germany), a canalised lowland river with a very flat floodplain. This river is highly polluted by nutrients from urban areas (Berlin) and a long chain of river lakes produces high concentrations of phytoplankton. Due to the high proportion of planktogenic detritus, it was hypothesised that greater quantities of nutrient-rich fine particulate organic matter (FPOM) would be deposited in floodplain waters located further from the main channel than has been reported for large rivers. The total nutrient, P-binding metal (Fe, Al, and Mn), organic and inorganic carbon (TOC, TIC) contents of the upper organic sediment layer (0–4 cm) were analysed in samples collected from 48 floodplain water and river sites. The sediment bulk density, calculated on the basis of dry mass content and loss on ignition, was used to characterise the waters according to the impact of the river current. The results showed that the variability of total phosphorus (TP) was best explained by the variability of total iron (TFe, R² = 0.52). The floodplain water sediments could clearly be separated into two groups on the basis of the sediment particle size composition, and of the element ratios TOC:TP, TN:TP, primarily TFe:TP. The sediments from impounded river sections and from mouth sections of backwaters (approx. 100–200 m) were characterised by a high proportion particles from the 0.1 to 0.5 mm size fraction and by homogeneous, low TFe:TP, TOC:TP and TN:TP ratios. Sediments from distal sections of backwaters and of oxbow lakes tended to exhibit high element ratios with much higher variability. These results were interpreted as a spatially limited impact of the river on the floodplain water sediments. Contrary to expectation, the phosphorus bound in river seston was predominantly and very homogeneously deposited in the impounded river and mouth sections of backwaters. This implies that the inundation of the floodplain waters during spring floods seems to have no important material impact on the sediments in waters of low hydrological connectivity with the River Havel.     

Rhythmic 24 h Variation of Core Body Temperature and Locomotor Activity in a Subterranean Rodent (Ctenomys aff. knighti), the Tuco-Tuco

The tuco-tuco Ctenomys aff. knighti is a subterranean rodent which inhabits a semi-arid area in Northwestern Argentina. Although they live in underground burrows where environmental cycles are attenuated, they display robust, 24 h locomotor activity rhythms that are synchronized by light/dark cycles, both in laboratory and field conditions. The underground environment also poses energetic challenges (e.g. high-energy demands of digging, hypoxia, high humidity, low food availability) that have motivated thermoregulation studies in several subterranean rodent species. By using chronobiological protocols, the present work aims to contribute towards these studies by exploring day-night variations of thermoregulatory functions in tuco-tucos, starting with body temperature and its temporal relationship to locomotor activity. Animals showed daily, 24 h body temperature rhythms that persisted even in constant darkness and temperature, synchronizing to a daily light/dark cycle, with highest values occurring during darkness hours. The range of oscillation of body temperature was slightly lower than those reported for similar-sized and dark-active rodents. Most rhythmic parameters, such as period and phase, did not change upon removal of the running wheel. Body temperature and locomotor activity rhythms were robustly associated in time. The former persisted even after removal of the acute effects of intense activity on body temperature by a statistical method. Finally, regression gradients between body temperature and activity were higher in the beginning of the night, suggesting day-night variation in thermal conductance and heat production. Consideration of these day-night variations in thermoregulatory processes is beneficial for further studies on thermoregulation and energetics of subterranean rodents.     

Responses to reversed NH3 and NH4+ gradients in a teleost (Ictalurus punctatus), an elasmobranch (Raja erinacea), and a crustacean (Callinectes sapidus):evidence for NH4+/H+ exchange in the teleost and the elasmobranch

Ammonia excretion rates of channel catfish, Ictalurus punctatus, little skate (Raja erinacea), and blue crab (Callinectes sapidus) were measured in experimental regimes which permitted simultaneous assessment of the partial pressure gradients for nonionized NH3 and the chemical concentration gradients of NH4+. Under conditions of low external ammonia, the average ammonia excretion was +295 microM kg-1 h-1 for catfish, +149 microM kg-1 h-1 for blue crabs, and +59 microM kg-1 h1 for skates with partial pressure gradients of +72.5 mu Torr, +413 mu Torr, and +24.4 mu Torr, respectively; and [NH4+] gradients of +189 microM l-1, +643 microM l-1, and +107 microM l-1 (positive indicating greater from inside to medium). When the external ammonia was increased to 1.15 mM l-1, both gradients were reversed, and the net ammonia movement was initially from the external water into all three species. In the catfish the inward movement ceased, however, and ammonia excretion eventually resumed in the face of reversed gradients of both NH3 partial pressure and [NH4+]. Unidirectional Na+ influx, indicative of a Na+/NH4+ exchange, did not increase. The ammonia data, changes in titratable acidity, and net apparent H+ efflux were all consistent with a linked extrusion of internal NH4+ for external H+. Incorporation of such an exchange into a computer simulation model of the ammonia equilibrium and exchange system duplicated the experimental data. Other hypotheses failed to match experimental data, or failed to predict internal ammonia levels lower than outside. In the crab, internal ammonia levels rose rapidly to concentrations and partial pressures above the external medium until excretion was reestablished, with no evidence of maintenance of a reversed gradient. In the skate, internal concentrations rose appreciably in the first hour and continued to rise for 6-8 h, with no resumption of ammonia excretion. The interspecies differences appear to be due at least partly to differences in ammonia permeability of the gills.