The routine differential leukocyte count vs automated differential counts

The clinical use of the proposed performance standards for differential leukocyte counts is determined by multiple factors. Their use must be considered according to the specific use of the differential count, the sources of variability in differential counting, the relation between specific use and sources of variability, the role of analytic errors in the detection of nonspecific changes, the use of qualitative vs quantitative data, the sensitivity and specificity of the routine eye count, the role of disease or specific cell prevalence in determination of predictive value, the effect on use of automated instruments for screening, and whether abnormal specimen flagging can be done. The routine eye-count differential method, as performed by a well-trained technologist or technician, seems to lack both sensitivity and specificity. Because of the magnitude of technique- and method-related and biologic sources of variability, the 100- or 200-cell eye-count differential method is not a good screening method for detection of hematologic illnesses, particularly those that are uncommon. The automated differential leukocyte instruments address many of the technique- and method-related errors and are thus able to equal or exceed the performance of the routine eye-count differential method.     

Liposomes with differential lipid components exert differential adjuvanticity in antigen-liposome conjugates via differential recognition by macrophages

We previously reported that liposomes having differential lipid components displayed differential adjuvant effects when antigen was coupled with liposomes via glutaraldehyde. In the present study, antigen-liposome conjugates prepared using liposomes having differential lipid components were added to the macrophage culture, and phagocytosis and the antigen digest of liposome-coupled antigen by macrophages were then investigated. Antigen presentation by macrophages to an antigen-specific T-cell clone was further investigated using the same conjugates. Antigen-liposome conjugates which induced higher levels of antibody production in vivo were recognized more often, and the liposome-coupled antigen was digested to a greater degree by macrophages than antigen-liposome conjugates which induced lower levels of antibody production. These results correlated closely with those regarding antigen presentation by macrophages; when antigen was coupled to liposomes showing higher adjuvant effect, macrophages cocultured with antigen-liposome conjugates activated antigen-specific T-cells at a higher degree. The concentration of OVA in the macrophage culture added as antigen-liposome conjugates was approximately 32 microg/mL. However, the extent of T-cell activation was almost equal to that when 800 microg/mL of soluble OVA was added to the culture. The results of the present study demonstrated that the adjuvant activity of liposomes observed primary in vivo correlated closely with the recognition of antigen-liposome conjugates and antigen presentation of liposome-coupled antigen by macrophages, suggesting that the adjuvant effects of liposomes are exerted at the beginning of the immune response, i.e., recognition of antigen by antigen-presenting cells.     

限制性酶切片段差异显示及其应用

差异显示技术(DD)-PCR是一种研究基因表达差异的重要而应用广泛的方法,传统的差异显示法由于在PCR时采用Poly（T）引物和随机引物而导致较高的假阳性率和产物的近Poly(A)非编码区的大量扩增。改进后的限制性酶切片段差异显示技术(RFDD)-PCR采用ToqI酶切双链cDNA,连上特殊设计的接头,再用经特殊设计的特异性配对于接头的引物来扩增,因此能重点扩增编码区并能极大地消除假阳性率。由于扩增时引物就带有荧光或放射性核素标记,还使得差异显示条带的检测更为方便、灵敏。本简要介绍了该法的原理、步骤、应用及其优缺点。     

A differential dilatometer

To obviate the difficulties resulting from partial solubility of membrane proteins in detergents or from the use of noxious solvent mixtures containing phenol or chloral, a simple procedure was devised for acrylamide gel electrophoresis of membrane proteins in 13 m formic acid. Polyacrylamide gels are equilibrated in 13 m formic acid and used in the electrophoresis assembly with 13 m formic acid as the electrolyte. Particulate proteinaceous preparations are dissolved in trifluoroacetic acid or in 24 m formic acid containing glycine to increase the density and to facilitate the solubilization of the protein. Protein samples (10 to 100 μg) migrate as polycations.     

mRNA差异显示

分离差异表达的基因是研究生命调节过程的重要手段,mRNA差异显示技术是一种能成功分离差异表达基因的方法,文章对该方法的基本原理、方法步骤及其应用作了较为详细的介绍。     

Multivariate analysis of microarray data: differential expression and differential connection

Background  

Typical analysis of microarray data ignores the correlation between gene expression values. In this paper we present a model for microarray data which specifically allows for correlation between genes. As a result we combine gene network ideas with linear models and differential expression.     

Multicolor fluorescent differential display

Differential display and DNA microarray have emerged as the two most popular methods for gene expression profiling. Here, we developed a multicolor fluorescent differential display (FDD) method that combines the virtues of both differential display in signal amplification and DNA microarray in signal analysis. As in DNA microarray, RNA samples being compared can be labeled with either a red or green fluorescent dye and displayed in a single lane, allowing convenient scoring and quantification of the differentially expressed messages. In addition, the multicolor FDD has a built-in signal proofreading capability that is achieved by labeling each RNA sample from a comparative study with both red and green fluorescent dyes followed by their reciprocal mixings in color. Thus, the multicolor FDD provides a platform upon which a sensitive and accurate gene expression profiling by differential display can be automated and digitally analyzed. It is envisioned that cDNAs generated by the multicolor FDD may also be used directly as probes for DNA microarray, allowing an integration of the two most widely used technologies for comprehensive analysis of gene expression.     

Weinberg's differential rule reconsidered

A central problem in research on twins is the estimation of the rates of monozygotic and dizygotic twin maternities. The estimation usually follows Weinberg's differential rule. According to this rule, the rate of dizygotic twinning is twice the rate of twin maternities in which the twins are of opposite sex. The monozygotic twinning rate is the difference between the rates of same-sex and opposite-sex twin sets. Weinberg's differential rule is implicitly based on the assumptions that the probability of a male equals the probability of a female and that the sexes in a dizygotic twin set are independent. Although Weinberg's differential rule has been the target of continuing discussions, the reliability of these assumptions has never been conclusively verified or rejected. In this study we present new variance formulas for the monozygotic and dizygotic twinning rates obtained using Weinberg's differential rule and stress that these new formulas have to be used. We analyze the accuracy of Weinberg's differential rule by considering alternative attempts. Especially, we build a general likelihood function and show that the maximum-likelihood estimates differ only slightly from the rates obtained using Weinberg's differential rule. In addition, our methods are applied to twinning data from Finland and Sweden. We compare our results with findings in the literature. In conclusion, our findings indicate that Weinberg's differential rule is rather robust and that despite its simplicity, it gives reliable results when official birth registers are analyzed.     

Subtractive differential display: a modified differential display technique for isolating differentially expressed genes

Differential display (DD) is a novel PCR-based technique, very commonly used to study differentially expressed genes at the mRNA level. In this paper we report a modified version of this technique that we have used to study the differences between the mRNA population from brain tissue of adult and old rats. We have modified the technique to enhance reproducibility and reduce false positives and redundancy. It is fast and does not require any expensive or uncommon reagent. We choose to call it as subtractive differential display as it is a differential display performed over subtracted mRNA population. We have used this protocol successfully to clone a number of age-related differentially expressed sequences from rat brain that need to be sequenced to establish the gene identity.     

A decade of differential display

It has been 10 years since the invention of differential display (DD), a conceptually simple methodology that allows the detection and identification of differentially expressed genes. In the past decade, the number of publications describing successful applications of DD has outnumbered those using any other competing methodologies, including subtractive hybridization, representational difference analysis, serial analysis of gene expression, and DNA microarrays. This review will provide a glimpse of the current progress made in DD technological development, refinement, and automation. Excellent examples of DD applications in studying a variety of biological problems, in such diverse biological systems as bacteria, yeast, flies, plants, and higher mammals, are presented to provide a roadmap for those who would like to pursue a fruitful gene "fishing" expedition. Some of the fundamental differences between DD and DNA microarrays are also discussed.     

Superparasitism as a differential game

Superparasitism refers to a female parasitoid laying an egg in a host already parasitized by a conspecific. In solitary species, only one offspring per host is expected to complete development, hence the game. Hosts are often clumped in patches and several females exploiting such an aggregate of resource make its state change over time, hence the dynamical character of the game. Two coupled questions arise: (i) Is it worth accepting a parasitized host? (ii) When to leave the host patch? Through these decisions (i) the competition for healthy hosts and (ii) the trade-off between leaving in quest of a better patch and staying to make the patch less profitable for other parasitoids (this is a way to lower superparasitism likely to occur after having left the patch) are addressed. The aim of this work is to characterize a strategy that would be evolutionarily relevant in such a situation, as it directly concerns females' reproductive success. Investigating a (synchronous) nonzero-sum two-player differential game allows us to characterize candidate dynamic evolutionarily stable policies in terms of both oviposition and patch-leaving decisions. For that matter, the game is (in the most part of the parameter space) completely solved if the probability that superparasitism succeeds is assumed to be close to one-half, a fair value under direct competition. The strategic equilibrium consists, for each females, in (i) superparasitizing consistently upon arrival on the patch, and (ii) leaving when the loss of fitness due to superparasitism likely to occur after its departure is reduced to zero. The competing females are thus expected to leave the patch as they arrived: synchronously. Superparasitism does not necessarily lead to a war of attrition.     

Protein biochips for differential profiling

Progress has been made in utilizing ProteinChip technology to profile and compare protein expression in normal and diseased states, particularly in the areas of cancer, infectious disease and toxicology. The past year has also seen the development of several novel chip types designed to analyze proteins in a fashion analogous to the array-based format of DNA microarrays. Some of these platforms may be used for differential profiling.     

Semi-supervised discovery of differential genes

Background  

Various statistical scores have been proposed for evaluating the significance of genes that may exhibit differential expression between two or more controlled conditions. However, in many clinical studies to detect clinical marker genes for example, the conditions have not necessarily been controlled well, thus condition labels are sometimes hard to obtain due to physical, financial, and time costs. In such a situation, we can consider an unsupervised case where labels are not available or a semi-supervised case where labels are available for a part of the whole sample set, rather than a well-studied supervised case where all samples have their labels.     

差异蛋白质组学的研究进展

差异蛋白质组是蛋白质组学研究的一个主要内容,其核心在于寻找某种特定臣寸素引起样本之间蛋白质组的差异,揭示并验证蛋白质组在生理或病理过程中的变化。进一步对蛋白质组差异信息分析后,理论上可以推断造成这种变化的原因。因此,对于临床上肿瘤预诊、药物靶标寻找、细胞调控分子的鉴别等有着极大的实际意义。差异蛋白质组研究要求可靠性和可重复性。因此,对于样本处理要求较高,激光微切割技术和高丰度蛋白去除技术的应用优化了样本处理方法。目前差异蛋白质组的主要研究方法仍是2-DE分离和MS鉴定联合应用,基于2-DE的2-DDIGE方法弥补了2-DE的弱点,更适用于差异蛋白质组研究。除2-DE技术外的其他几种技术手段,如多维液相色谱分离技术、ICAT技术、蛋白芯片技术等差异蛋白质组学研究技术可以作为2-DE技术的补充,甚至或替代技术。