Comparative Analysis of Esterase Activities of Human,Mouse, and Rat Blood

Acetylcholinesterase, butyrylcholinesterase, carboxylesterase, and paraoxonase activities in human, mouse, and rat blood were measured. The proportions of these enzymes activities differed significantly. In humans, the most significant were cholinesterase activities, while in rats and mice the contribution of carboxylesterase activity was the greatest. High arylesterase activity of paraoxonase was observed in all cases. Species-specific differences should be taken into consideration when carrying out preclinical trials on rodents for optimization of the pharmacokinetic characteristics of drugs containing complex ester groups.     

Automated Construction of High-Density Comparative Maps Between Rat, Human, and Mouse

Animal models have been used primarily as surrogates for humans, having similar disease-based phenotypes. Genomic organization also tends to be conserved between species, leading to the generation of comparative genome maps. The emergence of radiation hybrid (RH) maps, coupled with the large numbers of available Expressed Sequence Tags (ESTs), has revolutionized the way comparative maps can be built. We used publicly available rat, mouse, and human data to identify genes and ESTs with interspecies sequence identity (homology), identified their UniGene relationships, and incorporated their RH map positions to build integrated comparative maps with >2100 homologous UniGenes mapped in more than one species (approximately 6% of all mammalian genes). The generation of these maps is iterative and labor intensive; therefore, we developed a series of computer tools (not described here) based on our algorithm that identifies anchors between species and produces printable and on-line clickable comparative maps that link to a wide variety of useful tools and databases. The maps were constructed using sequence-based comparisons, thus creating "hooks" for further sequence-based annotation of human, mouse, and rat sequences. Currently, this map enables investigators to link the physiology of the rat with the genetics of the mouse and the clinical significance of the human.     

Immunohistochemical Examination of Novel Rat Monoclonal Antibodies against Mouse and Human Podoplanin

This study aims to develop new monoclonal antibodies (mAbs) against mouse and human podoplanin. Rats were immunized with synthetic peptides, corresponding to amino acids 38–51 of mouse podoplanin or human podoplanin which is 100% homologous to the same site of monkey podoplanin; anti-mouse podoplanin mAb PMab-1 (IgG_2a) and anti-human mAb NZ-1.2 (IgG_2a) were established. In immunocytochemistry, the mouse melanoma B16-F10 and mouse podoplanin (mPDPN)-expressed CHO transfectant were stained by PMab-1; human lymphatic endothelial cells (LEC) and human podoplanin (hPDPN)-expressed squamous cell carcinoma HSC3 transfectant, were stained by NZ-1.2. Western-blot analysis detected an about 40-kDa protein in CHO-mPDPN and B16-F10 by PMab-1, and in HSC3-hPDPN and LEC by NZ-1.2. In frozen sections, PMab-1 reacted with mouse kidney, pulmonary alveoli, pulmonary pleura, and salivary gland myoepithelial cells while NZ-1.2 reacted to the human salivary gland myoepithelial cells. The immunostaining of paraffin-embedded sections also showed the reaction of PMab-1 or NZ-1.2 to the mouse or monkey kidney glomerulus, pulmonary alveoli, and lung lymphatic vessels. These results indicate that the two novel rat mAbs to the mouse and human/monkey podoplanin are useful for Western-blot and immunostaining of somatic tissues on paraffin-embedded sections as well as frozen sections.     

Large-Scale Production and Partial Purification of Mouse Immune Interferon

Large-scale production of high-titered (10(2.2) to 10(4) U/ml) immune interferon (type II) was carried out in roller cultures of mouse spleen cells by using the T-cell mitogen staphylococcal enterotoxin A. Precipitation of 90% of this interferon by 55 to 80% saturated ammonium sulfate resulted in a 20-fold concentration and a two- to sixfold purification. After application of this interferon to either bovine serum albumin (BSA)-Affi-Gel 10 or hydroxylapatite columns, 100% of the interferon activity was recovered. By BSA-Affi-Gel 10 chromatography, 7% of the recovered activity was not bound, 45% was eluted with pH gradient 5 to 7, and 48% was eluted with 1 M NaCl. The pH- and salt-eluted interferons from the BSA-Affi-Gel 10 column were purified 62- and 390-fold, respectively, when compared with the starting materials. Rechromatography of the pH- and salt-eluted interferon peaks from the BSA-Affi-Gel 10 column did not alter their elution patterns. Stepwise elution of interferon from the BSA-Affi-Gel 10 columns with buffers of various pH and salt contents also resulted in greater than 300-fold purification. Specific activities of up to 2 x 10(5) U of interferon per mg of protein were attained with either elution procedure from BSA-Affi-Gel 10 columns. By hydroxylapatite chromatography, 5% of the recovered activity was not bound, 20% was eluted with a salt gradient, and 75% was eluted with 30% glycerin. Purification was 107- and 16-fold, respectively, for the two fractions. Ultrogel AcA 34 chromatography of the interferon resulted in two peaks of activity, a major one with a molecular weight of approximately 40,000 and a minor peak of molecular weight 70,000 to 90,000. Thus, by different types of chromatography, immune interferon was found to be heterogeneous.     

Clinical Genetics & Human Genome Variation: The 2008 Human Genome Variation Society Scientific Meeting

The annual scientific meeting of the Human Genome Variation Society (HGVS) was held on 11 November 2008, in Philadelphia, PA. The major theme of this meeting was “Clinical Genetics & Human Genome Variation.” For complex diseases, it is becoming evident that the contribution of most associated genetic variants to the disease process is small and, most likely, multiple variants are required to explain the predisposition and variation that is observed. As genome‐wide association studies (GWASs) identify variants that are associated with a disease, there is a need to determine if the associated variants are causative, or simply in genetic disequilibrium with the true functional variant. New methods are being devised to help classify these genetic variants as either functional or nonfunctional. As study populations increase in size, there is also a need for better‐constructed databases that can bring together the different genetic variants being identified, including SNPs, copy number variants (CNVs), and methylation differences, environmental risk factors, and the clinical information needed to construct useful phenotypes. These topics and others were discussed in this year's meeting. Hum Mutat 30, 852–856, 2009. © 2009 Wiley‐Liss, Inc.     

Ku and the Stability of the Genome

Ku proteins are associated with a variety of cellular processes such as repair of DNA-double-strand breaks, telomere maintenance and retrotransposition. In recent years, we have learned a lot about their cellular and molecular functions and it has turned out that Ku-dependent processes affect the stability of the genome, both positively and negatively, in several ways. This article gives an overview on the role of Ku in determining the shape of the genome.     

Human Autoantibodies Recognizing Human and Mouse Preimplantation Stages

PROBLEM: To find out whether autoantibodies against human preimplantation stages are present in some human sera and, if so, whether the antibodies could be capable to affect the egg development and/or to trigger an activation of the complement system at the procedures of assisted conception. METHODS: 1. Immunohistochemistry on blots of human preimplantation stages. 2. Immunohistochemistry on paraffin sections of human and mouse preimplantation stages. 3. Culture of mouse morulae to analyze complement activation. RESULTS: 1. Some human sera contained autoantibodies against human preimplantation stages. 2. Human-mouse cross-reacting antibodies against preimplantation stages occurred. 3. Immune complexes, formed on mouse preimplantation stages, activated the complement systems in egg cultures, resulting in a damaging of the eggs. CONCLUSION: The presence of natural autoantibodies to preimplantation stages may be associated with reproduction failure, caused by a direct effect by the autoantibodies and/or an activation of the uterine complement system by the immune complexes formed.     

Anti-Ia-reactive Cells in the Urinary Tract of Man, Guinea-Pig, Rat and Mouse

Tissues from the urinary tract of man, the guinea-pig, mouse and rat were investigated for la (HLA-DR) tissue distribution with an immunohistochemical staining technique using anti-Ia monoclonal antibodies. Human and guinea-pig tissues were also investigated with a rabbit anti-human HLA-DR immunoglobulin. Ia-antigen-expressing cells were demonstrated in bladder connective tissue of man, the guinea-pig, rat and mouse. In man and the guinea-pig anti-Ia-reactive cells were present also in the bladder epithelium. In the kidney, the Ia antigens seemed to be restricted to the endothelium in man, the guinea-pig and mouse, whereas in the rat, the kidney contained anti-Ia-reactive dendritic cells in the inter-stitium. The staining of mouse tissues with anti-I-A^k- and anti-I-E/C^k-specific monoclonal antibodies revealed that cells of different Ia phenotypes were differently distributed in the bladder connective tissue. In sections from the mouse kidney no difference in staining was seen with the various antibodies. In experiments with mouse bone-marrow chimeras the anti-Ia-reactive cells in mouse bladder were shown to be of bone-marrow origin. In humans epithelial Ia-expressing cells could be isolated from the urine. Such cells might constitute a source of anti-Ia-reactive epithelial cells for future functional studies.     

快速低成本全基因组DNA测序技术

人类个体的全基因组序列信息，是最为重要的生物医学信息，是实现未来个体化医疗的基础；实现个体全基因组DNA序列的快速低成本检测，是人类基因组计划完成后又一个重要的技术挑战。对该技术当前的发展现状进行回顾和分析，预计在不长的时间内人类个体全基因组的测序成本能够降低到1万元人民币以下。     

Deep Brain Stimulation, Brain Maps and Personalized Medicine: Lessons from the Human Genome Project

Although the appellation of personalized medicine is generally attributed to advanced therapeutics in molecular medicine, deep brain stimulation (DBS) can also be so categorized. Like its medical counterpart, DBS is a highly personalized intervention that needs to be tailored to a patient’s individual anatomy. And because of this, DBS like more conventional personalized medicine, can be highly specific where the object of care is an N = 1. But that is where the similarities end. Besides their differing medical and surgical provenances, these two varieties of personalized medicine have had strikingly different impacts. The molecular variant, though of a more recent vintage has thrived and is experiencing explosive growth, while DBS still struggles to find a sustainable therapeutic niche. Despite its promise, and success as a vetted treatment for drug resistant Parkinson’s Disease, DBS has lagged in broadening its development, often encountering regulatory hurdles and financial barriers necessary to mount an adequate number of quality trials. In this paper we will consider why DBS—or better yet neuromodulation—has encountered these challenges and contrast this experience with the more successful advance of personalized medicine. We will suggest that personalized medicine and DBS’s differential performance can be explained as a matter of timing and complexity. We believe that DBS has struggled because it has been a journey of scientific exploration conducted without a map. In contrast to molecular personalized medicine which followed the mapping of the human genome and the Human Genome Project, DBS preceded plans for the mapping of the human brain. We believe that this sequence has given personalized medicine a distinct advantage and that the fullest potential of DBS will be realized both as a cartographical or electrophysiological probe and as a modality of personalized medicine.     

Increased Susceptibility of Periodate-Treated Tumour Cells to Human but Not to Mouse or Rat Natural Killer Cells

Treatment of intact cells in the cold with low concentrations (1 mM) of sodium meta periodate (PI) selectively oxidizes the surface-exposed sialic acid residues to the corresponding aldehydes. Such treated tumour cells show greatly enhanced sensitivity to lysis by fresh human NK cells but not to mouse or rat NK cells. Reduction of the PI-treated cells with sodium borohydride (NaBH4) reduced their NK sensitivity to that of untreated cells. In target conjugate formation assays PI-treated tumour cells displayed a higher binding capacity than control cells or PI+NaBH4-treated cells to both mouse and human effector cells. Neuraminidase treatment of K562 and Molt-4 increased target susceptibility to human NK cells but not to mouse, whereas the susceptibility of Yac-1 cells was left unchanged using both human and mouse effector cells. The same pattern of reactivity is shown in the target binding assay. These findings indicate that subtle molecular changes in the surface-exposed carbohydrates of target cells might have a fundamental impact on their sensitivity to lysis by NK cells from certain species, and that in cross species effector-target combinations a higher binding capacity is not sufficient for increased lysis to occur.     

Mouse Virulence of Human, Systemic, Pathogenic Escherichia coli Strains

Strains of Escherichia coli isolated from systemic infections of humans exhibited marked intraperitoneal and intracerebral virulence for mice. This marked virulence was not exhibited by enteropathogenic strains.