PCR-SSCP--rapid and easy detection of DNA-sequence changes]

PCR-SSCP analysis is a rapid, simple and sensitive technique for detection of various mutations, including single nucleotide substitutions, insertions and deletions, in PCR-amplified DNA fragments. Here I review, the principle and sensitivity of the technique, and also show how this technique has been used in clinical and basic medical sciences.     

State-dependent life-history theory and its implications for optimal clutch size

Summary Life-history theory is usually based on an animal's age or size. McNamara describes a general technique for finding the optimal life-history when an organism's strategy is allowed to depend on other aspects of its state. In this paper we describe the technique in the context of previous work in life-history theory and discuss how it can be used to look at decisions on a finer time scale than the usual annual decisions. We show how it can be used to model optimal clutch size when there is a trade-off between number and quality of offspring. It is shown that the optimal clutch size is typically less than the most productive clutch size. Measuring the value of a clutch in terms of the number of offspring that survive to breed or even the number of grandchildren that survive to breed may give misleading results.     

Determination of Photoreceptor Cell Spectral Sensitivity in an Insect Model from In Vivo Intracellular Recordings

Intracellular recording is a powerful technique used to determine how a single cell may respond to a given stimulus. In vision research, intracellular recording has historically been a common technique used to study sensitivities of individual photoreceptor cells to different light stimuli that is still being used today. However, there remains a dearth of detailed methodology in the literature for researchers wishing to replicate intracellular recording experiments in the eye. Here we present the insect as a model for examining eye physiology more generally. Insect photoreceptor cells are located near the surface of the eye and are therefore easy to reach, and many of the mechanisms involved in vision are conserved across animal phyla. We describe the basic procedure for in vivo intracellular recording of photoreceptor cells in the eye of a butterfly, with the goal of making this technique more accessible to researchers with little prior experience in electrophysiology. We introduce the basic equipment needed, how to prepare a live butterfly for recording, how to insert a glass microelectrode into a single cell, and finally the recording procedure itself. We also explain the basic analysis of raw response data for determining spectral sensitivity of individual cell types. Although our protocol focuses on determining spectral sensitivity, other stimuli (e.g., polarized light) and variations of the method are applicable to this setup.     

Advances in the application of scanning electrochemical microscopy to bioanalytical systems

Scanning electrochemical microscopy (SECM) is a powerful surface characterisation technique that allows for the electrochemical profiling of surfaces with sub micrometer resolution. While SECM has been most widely used to electrochemically study and profile non-biological surfaces and processes, the technique has in recent years, been increasingly used for the study of biological systems - and this is the focus of this review. An overview of SECM and how the technique may be applied to the study of biological systems will first be given. SECM and its application to the study of cells, enzymes and DNA will each be considered in detail. The review will conclude with a discussion of future directions and scope for further developments and applications.     

Alternative approach to modal gait analysis through the Karhunen-Loève decomposition: An application in the sagittal plane

This article uses the Karhunen-Loève decomposition (KL) technique, also known as proper orthogonal decomposition (POD) or principal component analysis (PCA), to introduce the concept of gait mode, which can be used as a tool to identify gait differences among subjects or groups and to approximate gait curves. The KL is a statistical pattern analysis technique for finding dominant structures in an ensemble of data. This technique can be used to decompose a spatiotemporal signal into time-independent, orthogonal, spatial components and time-dependent amplitudes. This study demonstrates the existence of a common gait mode through the analysis of the kinematics of 57 young, healthy subjects, and how they can be used to identify different walking patterns, for instance differentiating male and female subjects.     

On predicting extinction in simple population models. I. Stochastic linearization

The technique of stochastic linearization derived by Bartlett is used to give an approximate solution to each of three stochastic models of predation. By defining the existence of a quasi-stationary equilibrium distribution of population sizes, the approximate variances and covariances of the joint distribution of population sizes are calculated. The results are used to predict whether prey or predators are likely to become extinct first and the predictions are tested against simulation data. The linearization technique is a good predictor of the outcome of extinction but not of how long it takes.     

Biointeraction analysis of immobilized antibodies and related agents by high-performance immunoaffinity chromatography

A method is described based on high-performance immunoaffinity chromatography for examining the interactions of immobilized antibodies or related binding agents with their targets. It is shown how this method can be used to obtain information on the binding, elution and regeneration kinetics of immobilized binding agents, such as those used with immunoaffinity supports. The theory behind this approach is briefly described and it is demonstrated how both the kinetic and thermodynamic properties of a biointeraction can be determined experimentally through this method. Several applications are used to illustrate this technique, including antibody-antigen interactions and the binding of aptamers with their targets in the presence of silica-based supports. The same approach can be adapted for use with other types of targets, binding agents and support materials.     

Good practice in head and neck fine needle aspiration cytology as assessed by CUSUM

Providing data as evidence of good practice is becoming imperative to meet the demands of professional revalidation and clinical governance. Sensitivity and specificity are common performance measures in fine needle aspiration (FNA) but are vulnerable to discordant analytical methods. We introduce a CUSUM technique and show how it may be used to show attainment and maintenance of proficiency in head and neck (H&N) FNA. In addition, we show how it can be used to compare practices and demonstrate different performance for FNAs from different tissues; a fact that must be recognized by anyone devising minimum performance values.     

Advantages to mutagenesis techniques generating populations containing the complete spectrum of single codon changes

The limitations of current mutagenesis techniques are analyzed in terms of the number and kinds of codon changes they make and in terms of the population size needed to produce all single or multiple amino acid variants. It is shown how a technique that can alter a single codon of a gene, producing all possible variant codons without affecting the rest of the gene, has certain advantages, if it can be used at each place in the gene in one experiment. Such a technique has advantages when the goals are to understand: (1) how specific structural alterations in a mutant protein cause it to function in a different but specific way, (2) how to predict which amino acids in a protein contact or interact with each other, and (3) why a protein is more or less sensitive to mutational disruption, depending upon the specific mutation. This is because it would generate the maximum number of (1) mutant proteins with different functions, (2) intracistronic suppressor for any starting mutation, and (3) random amino acid substitutions at random places. Furthermore, such a technique could produce useful variants more quickly and on a smaller scale than either evolution or current methods.     

活体动物全细胞记录技术及其应用

活体动物全细胞记录技术不仅可以用于研究感觉系统对自然刺激（如视觉系统的光刺激、听觉系统的声音刺激等）反应的特性和规律,还可以较准确地记录细胞的突触电位（包括阈下反应）,实现EPSP和IPSP的相对分离,并实现活体细胞内灌流,从而进一步研究感觉信息的处理机制。本文较为详细地介绍了在活体动物上进行全细胞记录的方法,包括一些技术细节和关键仪器设备的选取原则,举例说明了该技术在视觉系统研究和体感系统研究中的应用,并讨论了这一方法在神经科学中的应用前景。     

Rhinoplasty in the African-American patient

Because of the increasing popularity of rhinoplasty in the African-American patient, we delineate how a rhinoplasty surgeon can perform this challenging technique to obtain uniform and consistent results. First, we address how one can appreciate and analyze the various aesthetic concepts of beauty and the unique anatomic characteristics of the African-American nose. Second, we present a pragmatic, systematic analysis of the African-American nose. Last, we describe the techniques consistently used to modify the African-American nose while achieving or maintaining facial harmony using the open approach to rhinoplasty. Specific case analyses are presented to demonstrate utilization of the technique.     

Nonparametric variable importance assessment using machine learning techniques

In a regression setting, it is often of interest to quantify the importance of various features in predicting the response. Commonly, the variable importance measure used is determined by the regression technique employed. For this reason, practitioners often only resort to one of a few regression techniques for which a variable importance measure is naturally defined. Unfortunately, these regression techniques are often suboptimal for predicting the response. Additionally, because the variable importance measures native to different regression techniques generally have a different interpretation, comparisons across techniques can be difficult. In this work, we study a variable importance measure that can be used with any regression technique, and whose interpretation is agnostic to the technique used. This measure is a property of the true data‐generating mechanism. Specifically, we discuss a generalization of the analysis of variance variable importance measure and discuss how it facilitates the use of machine learning techniques to flexibly estimate the variable importance of a single feature or group of features. The importance of each feature or group of features in the data can then be described individually, using this measure. We describe how to construct an efficient estimator of this measure as well as a valid confidence interval. Through simulations, we show that our proposal has good practical operating characteristics, and we illustrate its use with data from a study of risk factors for cardiovascular disease in South Africa.     

Anti-inflammatory action of therapeutic and low-frequency ultrasound on the inflammatory process model on rats

The results of experimental investigations of the anti-inflammatory effect of ultrasound on the inflammatory process on a rat model of carrageenan-induced edema of the hind limb are given. The technique used in the experiment showing ultrasound exposure parameters and how to control the characteristics of inflammatory process is described. The anti-inflammatory effect of ultrasound is established.     

Research report: The study of ethnic alignment: A new technique and an application in Malaysia

In some circumstances, like driving a motor vehicle, group membership is irrelevant. In others, individual interest motivates group alignment. Sometimes choices have to be made between individual responses and group alignment, or between alignment on the basis of class, or ethnicity, or religion. The authors describe a technique used to assess the strength of ethnic loyalty relative to self‐interest and personal obligation among an urban sample in Malaysia. Predictions of how a representative Malay would react in situations of value conflict made possible the measurement of pluralistic ignorance. The technique could be used elsewhere. It could be used to generate predictions about trends in the strengthening, maintenance or weakening of ethnic boundaries.     

The Analysis of Life Table and Follow-up Data with Covariates Using Poisson Regression Model

The Poisson regression model for the analysis of life table and follow-up data with covariates is presented. An example is presented to show how this technique can be used to construct a parsimonious model which describes a set of survival data. All parameters in the model, the hazard and survival functions are estimated by maximum likelihood.     

Methods in molecular cardiology: DNA sequencing

Sequencing is one the major breakthroughs in molecular cardiology. The development of this technique has made it possible to determine the exact order of the nucleotides in DNA. The exact order is relevant for the formation of proteins, through the genetic code. Sequencing is even more important for the identification of genetic variation and disease-causing mutations. The elucidation of the human genome is based on the continuous improvement of this technique, reducing the cost and increasing efficiency. Initially, complex chemical reactions were performed using isotopes to unravel the base sequence in genes. Nowadays, fluorescent capillary-based techniques are available to determine the genetic information. Here, the historical development of the technique is described. In addition, examples are provided on how sequencing is used in clinical medicine.     

Ups and downs of protein crystallization: studies of protein crystals by high-performance capillary electrophoresis

High-performance capillary electrophoresis is a high-technology micro-separation method. Short run time, full automation and minute amounts of sample make it a very attractive technique. In this report we describe studies of protein crystals by capillary electrophoresis. We show how high-performance capillary electrophoresis can be used effectively for rapid evaluation and examination of the protein solution used for crystallization, the protein crystals (solubilized) and surrounding mother liquor. With coated capillaries, the runs were reproducible and disturbing effects, such as electroendosmosis and interaction of the proteins with the capillary wall, were suppressed efficiently. We recommend this new technique as a powerful and routine companion to protein crystallography.     

Determining the Reactivity and Titre of Serum using a Haemagglutination Assay

Haemagglutination is a specific form of agglutination and is used when antibodies bind to red blood cells, which act as a particulate antigen. Red blood cells are particularly useful targets as they are readily available and agglutination is observable using the naked eye. This technique is commonly used to determine the titre of an antibody (Ab), for blood grouping and viral quantification. In this video, the steps involved in preparing and performing a haemagglutination assay is demonstrated using antibodies specific to blood group A-antigens added to red blood cells (Revercells). The antiserum is serially diluted in a 96 well U-bottom microtitre tray, to which is added a suspension of Revercells. The samples are mixed and then incubated at 37°C for 60 minutes. After this time, the samples can then be easily scored for ve, +ve and intermediate (-/+) haemagglutination reactions. This approach allows for the reactivity and titre of a serum sample to be assessed using a rapid and simple technique. The video will cover the theory behind the assay, how the results are read and interpreted, how the titre is determined, how the assay can be modified and any issues associated with the use of this technique.Open in a separate windowClick here to view.^{(20M, flv)}     

Calculation of the Volterra kernels of non-linear dynamic systems using an artificial neural network

The Volterra series is a well-known method of describing non-linear dynamic systems. A major limitation of this technique is the difficulty involved in the calculation of the kernels. More recently, artificial neural networks have been used to produce black box models of non-linear dynamic systems. In this paper we show how a certain class of artificial neural networks are equivalent to Volterra series and give the equation for the nth order Volterra kernel in terms of the internal parameters of the network. The technique is then illustrated using a specific non-linear system. The kernels obtained by the method described in the paper are compared with those obtained by a Toeplitz matrix inversion technique. Received: 4 June 1993/Accepted in revised form: 2 March 1994     

滚环扩增信号放大技术在生物检测中应用的研究进展

滚环扩增(Rolling circle amplification,RCA)是一种快速、灵敏且恒温的单链DNA(Single-stranded DNA,ssDNA)扩增技术,与染色或探针联用可实现检测信号的放大,在生物检测等方面得到广泛的应用。文中对RCA的构建方法进行了简介,综述了近几年其在致病菌、核酸肿瘤标记物、蛋白质、生物小分子和病毒等检测中的研究进展,并对其未来的发展趋势进行了展望。