The Enamel Microstructure of the Early Eocene Pantodont Coryphodonand the Nature of the Zigzag Enamel

An additional method for the investigation of the microstructure of enamel is described using the teeth of Coryphodon, Uintatherium, Entelodon, and Crocuta. Under low magnification natural surfaces or sections of teeth display details of the enamel microstructure when the light guide effect of prisms is used. Under the same low magnification even more details were obtained from sputter-coated surfaces of sections. The method is of particular significance for the investigation of large teeth with thick enamels when structures are somewhat irregular. The new method provides a better general survey, where scanning electron microscope images often show confusing details. The enamel of Coryphodon shows oblique lines of nested chevrons that are similar, to some degree, to the zigzag enamel in Crocuta, but a distinct asymmetry between ascending and descending lineaments was observed. This specific Coryphodon -enamel was also found in Uintatherium and Entelodon. This enamel type, which evolved several times in parallel, cannot be attributed to a specific diet, but must be regarded as one of the several ways to strengthen the enamel against breakage.     

Comparison of the relative and k0 methods for the standardization of NAA with stable low-flux reactors

Thek ₀ standardization method was Actapted for NAA with stable low-flux reactors where flux monitors are not needed. The modifiedk ₀ method offers the convenience of the use of libraries of sensitivity constants. It was compared to the relative method for 52 elements using a SLOWPOKE reactor and 6 counting geometries. The sensitivity constants determined fromk ₀ values were found to be as accurate or more accurate than those measured with standards. NAA with this modifiedk ₀ method should be accurate to 3% for light elements and 5% for heavy elements.     

A qPCR method to characterize the sex type of the cell strains from rats

A simple and fast method was established to identify the sex types of the rat-derived cell strains. The single copy X-chromosome-linked gene AR and the single copy Y-chromosome-linked gene Sry were both detected with qPCR for the rat genomic DNA sample and the AR/Sry ratio was calculated. According to the law of the AR/Sry ratio, a new method to identify the sex types of the rat-derived cell strains was developed. The new assay was proved effective. The new assay showed advantages over the traditional sex type identification PCR methods, which detected only the Sry gene. Moreover, the new method was used to identify the sex types of two rat-derived cell strains unknown for the sex types and the results were confirmed with the in situ hybridization. Finally, the problem of the cross contamination between the female and the male samples was addressed and discussed extensively.     

Laboratory culture of the mitesVarroa jacobsoni andTropilaelaps clareae

A laboratory method was developed for rearing the mitesVarroa jacobsoni Oudemans andTropilaelaps clareae Delfinado and Baker to the adult stage. The method consists of infesting larvae ofApis cerana Fabricius andA. mellifera L. with mites in ELISA microtiter racks.     

On the Estimation of the Nearest Neighbour Distribution,G(t), for Point Processes

A new method is proposed for estimating G(t), the distribution function of the distance from an object to its nearest neighbour in a spatial point process. The new method makes more complete use of the information available and has a smaller mean squared error than that of the existing alternatives. The method appears equally effective with random, clustered and regular patterns.     

Estimation of the stage-specific survival rate in the insect population with overlapping stages

An application ofHokyo andKiritani 's method (1967) was attempted to estimate the stage specific survival rates of the population with overlapping stages. This method can be written as follows assuming a constant daily survival rate (K) throughout the life: where, and F refer respectively to the total incidence of ith instar nymphs and that of individuals after ith instar inclusive, and α_i refers to the developmental period of ith instar. Application of this model to caged and natural populations of the southern green stink bug, Nezara viridula, was made to test its validity. The estimates of the initial number of successive stages obtained from the present method were compared with those fromRichards andWaloff 's method (1954) for the caged populations of 1st, 2nd and 3rd generations. The superiority of the present method to theRichards andWaloff 's in estimating adult numbers was shown in all the generations examined. When different daily survival rates are involved in the course of population decrease, application of the revised method proposed byHokyo andKiritani (1967), gives much reliable estimate as compared with one before correction. The present method is useful in constructing life table of such species as scale insects which complete their life cycle within a defined space, but their successive stages overlap considerably.     

Enantiopurity Determination of the Enantiomers of the Triple Reuptake Inhibitor Indatraline

The present study describes the development of two approaches for the determination of the enantiopurity of both enantiomers of indatraline. Initially, a method was developed using different chiral solvating agents (CSAs) for diastereomeric discrimination regarding signal separation in ¹H nuclear magnetic resonance (NMR) spectroscopy, revealing MTPA as a promising choice for the differentiation of the indatraline enantiomers. This CSA was also tested for its ideal molar ratio, temperature, and solvent. Optimized conditions could be achieved that made determination of enantiopurity for (1R,3S)‐indatraline up to 98.9% enantiomeric excess (ee) possible. To quantify even higher enantiopurities, a high‐performance liquid chromatography (HPLC) method based on a modified β‐cyclodextrine phase was established. The influence of buffer type, concentration, pH value, percentage and kind of organic modifier, temperature, injection volume as well as sample solvent on chromatographic parameters was investigated. Afterwards, the reliability of the established HPLC method was demonstrated by validation according to the ICH guideline Q2(R1) regarding specificity, accuracy, precision, linearity, and quantitation limit. The developed method proved to be strictly linear within a concentration range of 1.25–1000 μM for the (1R,3S)‐enantiomer and 1.25‐750 μM for its mirror image that enables a reliable determination of enantiopurities up to 99.75% ee for the (1R,3S)‐enantiomer and up to 99.67% ee for the (1S,3R)‐enantiomer. Chirality 25:923–933, 2013. © 2013 Wiley Periodicals, Inc.     

Reliability of the resorcinol method for identifying isomorphic phases in the Gigartinaceae (Rhodophyta)

The Gigartinaceae are economically important because the carrageenans in this family are used for a variety of purposes including food stabilizers, food substitutes, pharmaceutical applications and cosmetics. The resorcinol method takes advantage of the different carrageenans found in life history phases in this family to identify nonreproductive phases, and consequently, a better understanding of the biology of this economically important family is possible. This study investigates sources of variability that could affect the accuracy of the resorcinol method in identifying phases within the Gigartinaceae. Vegetative disks of Iridaea splendens from different areas of the blade and disks containing reproductive structures all reacted consistently to the resorcinol reagent. The minimum size and testing conditions required to reliably identify phases at the germling stage of I. splendens were determined. Phases of I. splendens cannot be identified if blades are stored in 3% formaldehyde in seawater. Phases of I. splendens, I. lineare, I. heterocarpa, Rhodoglossum californicum and R. affine can be identified reliably at different times of the year. Manipulations of the resorcinol method improved its accuracy in identifying phases of Iridaea cornucopiae and Gigartina exasperata but the high percent of G. exasperata misidentified demonstrates that it cannot be assumed that the current resorcinol method works for all taxa in the Gigartinaceae.     

医院科室工作量标准化及其与职工数的相关性研究

目的 建立科室工作量标准化方法及科室职工数的相关联变量。方法 调研临床科室、访谈医务人员,分析医务人员关于工作量量化考核的意见和建议;梳理样本医院内部绩效工资分配测算方法在工作量标准化上的问题。对科室数据进行模拟测算分析。结果 建立基于科室日均工作负担的科室工作量标准化测算方法,科室日均工作负担与科室职工数显著相关。结论 标准化的科室日均工作负担改进科室绩效工资测算的合理性。     

The Chemical Potential of Dense Liquids by the Coupled Test Particle Method

Abstract

A new method for chemical potential estimation is proposed which is based on the coupled particle approach. The coupled particle method defines an attractive solution to the weighting function problem in umbrella sampling, bridging the gap between f and g distributions at high density. A way of eliminating the origin singularity is suggested, which is similar in spirit to the restricted umbrella sampling of Shing and Gubbins, but which is based on geometric rather than energetic criteria.

The method is illustrated on the Lennard-Jones system up to a reduced density ρ? = 1.1 along the isotherm T? = 1.2 and results are compared with the test particle insertion method and empirical equations of state. The new method is particularly useful at high liquid densities where it is superior to the other methods relying on the degree of overlap of f and g distributions. It gives reliable estimates of the chemical potential in the whole range of liquid densities.     

The Value of the Smear Method for Plant Cytology

The smear method of plant cytology as described by Taylor, presented difficulties in obtaining an adequate stain with the iron-alum haematoxylin combination. The writer has largely avoided those difficulties by curtailing the times of mordanting and of staining. The smear method has been applied by the writer in the study of microsporogenesis in the monocotyledonous genera Tradescantia and Rhoeo and in the dicotyledonous genus Podophyllum. Due to the rapidity with which permanent preparations can be completed by the smear method, there is presented a valuable means for securing a more critical evaluation of the available preservatives.     

Quantifying the colony shape of the Montastraea annularis species complex

The reef coral Montastraea annularis has been used in a wide range of investigations. Recently, it has been recognized as a complex of three species based on field observations of the variation in colony shape. These observations have also been confirmed by molecular methods as well as morphometrics on individual corallites in the colonies. This paper presents a new quantitative method for measuring overall colony shape based on geostatistics. Seventeen colonies collected from San Blas, Panama in 1995 and 1996 were examined using the Polhemus 3SPACE FASTRAK system to construct three-dimensional coordinates of the center of several hundred corallites in each colony. This method measures the larger bumps or “ridges” as well as the smaller bumps or “lumps” of the colonies. Variograms were then calculated for all the specimens and the lag distances and the values of the variogram were used in a multivariate statistical analysis. Overall, this method indicated that M. annularis and Montastaea faveolata overlap in their relative amount of bumpiness in colony shape while Montastraea franksi is distinct from the other two species. This method has implications for both the modern and fossil record of Montastraea as well as other organisms with similar shapes.     

Detecting Surfactant-producing Microorganisms by the Drop-collapse Test

Summary A rapid method, ’drop-collapse’, was used for screening biosurfactant production by Pseudomonas aeruginosa, Bacillus subtilis, Candida albicans and Phanerochaete chrysosporium liquid cultures. Before measuring the total biosurfactant, the drop-collapse method was used in order to detect rhamnolipid presence in the culture broths. The method was performed in a microwell plate; the polystyrene platform with small wells. If the culture broth contained biosurfactant, the droplets of the broth in the oil-coated wells collapsed. If not, there was no change in the shape of the droplets. Pseudomonas aeruginosa and Bacillus subtilis culture supernatants showed spreading movement, meaning that they produced biosurfactants. However, Candida albicans and Phanerochaete chrysosporium supernatants remained beaded, meaning they did not produce any type of microbial surfactant.     

Age changes of the carpal bones in macaques: Application of the fourier analysis on the radiographs

A method based on the Fourier analysis is proposed, which describes and analyzes the contour morphology of carpal bones by separating morphology into factors of shape and size. Here, “size” refers to the average diameter of the contour. The “shape” is expressed byshape factors which are derived from the Fourier series and the “shape” of a monkey is expressed by ashape index which is calculated fromshape factors. The age change in the morphology of the lunate and capitate ofMacaca fuscata fuscata was analyzed by this method. The development of “shape” approximately completes by 3 years of age, whereas increase in “size” begins its spurt at that age as do body weight and anterior trunk length. By applying this method to other macaque species, it was found thatM. mulatta, M. f. yakui andM. cyclopis exhibit similar patterns of growth and development of carpal bones to those ofM. f. fuscata. Patterns found inM. fascicularis differ in that its bones develop faster than in the other macaques with respect to the “shape,” but remain small with respect to the “size.”     

Theory and practice of the analytical centrifugation of an active substrate-enzyme complex

The active enzyme centrifugal-ion (AEC) method presented here permits the hydrodynamic study of active enzyme–substrate(s) complexes in solutions containing micro-gram amounts of the studied enzyme, even if the enzyme preparation is very impure. The AEC method can he used only when the specific enzymatie reaction can be measured directly in a spectrophotometer. The general equations relevant to the method and their solutions are presented in detail. Their use requires some numerical calculations. A practical summary of the AEC method is given, and the precision of the measured values of the sedimentation and diffusion coefficients is discussed.     

Use of Quantitative Real-Time PCR to Investigate the Dynamics of the Red Tide Dinoflagellate Lingulodinium polyedrum

A new method based on quantitative real-time polymerase chain reaction (qPCR) was developed and applied to quantify the red tide dinoflagellate Lingulodinium polyedrum in natural seawater samples and in laboratory cultures. The method uses a Molecular Beacon™ approach to target a species-specific region of the small subunit ribosomal RNA gene. The accuracy of the method was verified by microscopical counts using cultures of the dinoflagellate isolated from coastal waters near Los Angeles, CA, and with natural water samples spiked with cultured L. polyedrum. The method was applied to document the pattern and timing of vertical migration by the dinoflagellate in a 2-m water column on an 11:13 h light/dark photoperiod established in the laboratory. Positive phototaxis of L. polyedrum resulted in dense aggregations of the dinoflagellate within the top few centimeters of the water column during the light period. This pattern of distribution was readily established by both methods, although abundances of L. polyedrum determined using qPCR were higher than abundances determined by microscopy in the morning and lower in the afternoon and evening. These differences may have been a consequence of variability in the DNA content per cell because of synchrony of cell division. Counts using both methods to analyze natural samples collected from coastal waters in the Long Beach–Los Angeles area and adjacent San Pedro Channel were in close agreement. However, the qPCR method exhibited greater sensitivity than the microscopical method when L. polyedrum was present at low abundances, and qPCR had a much higher rate of sample throughput than microscopy. The development of this new approach for enumerating L. polyedrum provides a useful tool for studying the ecology of this important red tide species.     

Determination of the Activity of Immobilized Biocatalysts in the Case of Poorly Soluble Substrates: The Example of D-5Phenylhydantoinase

The present communication describes the determination of activity of immobilized biocatalysts, from progress curves, in the case of a poorly soluble substrate. Computer simulation is used for the comparison of five well known methods for the determination of initial velocity of the reaction with the modified method of smoothing experimental data by cubic spline. The computer simulation data show that, in the case of poorly soluble substrate, it is expedient to use the method of data linearization in the co-ordinate system p/t versus p and method of splines.     

Comments on the approximate solution of the diffusion equation: I

The approximation method of N. Rashevsky is discussed and reviewed. It is shown that in addition to theexplicit assumptions and approximations there is involved the assumption that the rate of metabolism is the same at every point in the cell and that theaverage rate of metabolism is different from zero. An expression is given for the error in the approximate method when the rate of metabolism is any function of the concentration. It is also shown that a solution in theform of that obtained by the approximate method is not possible if the generalized laws of diffusion are assumed to apply. A portion of this work was performed while the author was a research participant, Oak Ridge Institute of Nuclear Studies, assigned to the Mathematics Panel, Oak Ridge National Laboratory.     

Comments on the approximate solution of the diffusion equation: II

On the basis of a previous general formulation (Bull. Math. Biophysics,15, 21–29, 1953a) a discussion is given of the error in the approximation method of N. Rashevsky. This error, inherent in the method when the metabolic rate is different at each point in the cell, is discussed in detail and numerical values are presented for two particular cases: the rate proportional to the concentration and the rate a prescribed function of the spatial coordinates. It is shown that the formulation for the first case also applies to several other cases, that the error is negligible provided the rate is sufficiently small, and that the error is fairly sensitive to the cell size. If the rate depends upon the coordinatesalone a small rate is not sufficient to insure a negligible error. The relations between the exact method, the standard approximate method, an earlier approximate method (Physics,7 260, 1936), and a more recent refinement (Bull. Math. Biophysics,10, 201, 1948) of the standard method are discussed. This work was performed while the author was a research participant, Oak Ridge Institute of Nuclear Studies, assigned to the Mathematics Panel, Oak Ridge National Laboratory.     

Inter-Alu-like species-specific sequences in the Saccharum complex

Alu sequences constitute the most abundant family of short interspersed nuclear elements, SINEs, in the primate genome. The Alu-PCR method, which consists of amplification between Alu sequences, is usually applied in human genetics to provide polymorphic markers. Here we report the presence of Alu-like sequences in sugarcane and related species by applying the Alu-PCR-like method. Amplifications using a PCR primer defined in conserved regions of Alu human sequences lead to specific complex multiband profiles in all the Saccharum and related genera clones surveyed. The isolation and characterisation of the amplified genus-specific inter-Alu-like fragments allowed us to isolate repeated sequences that are specific for different genera of the Saccharum complex: MsCIR2 from Miscanthus, EaCIR6 and EaCIR7 from Erianthus, and SrCIR2 from Saccharum. Two PCR diagnostic tests were developed from the inter-Alu-like sequences MsCIR2 and EaCIR6, and proved efficient in identifying intergeneric hybrids Saccharum×Miscanthus or Saccharum×Erianthus, respectively. The present study illustrates how the Alu-PCR-like method could help investigating the origin of amphiploid species and monitoring introgression in plants. Received: 7 March 1999 / Accepted: 25 March 1999