Implementation of a Multi-sampling-frequency System for RR-interval Timing and Blood Pressure Detection

INTRODUCTION Researchers study on cardiac electrophysiology and haemodynamics for more effective methods of di-agnosis and treatment. During these studies, howto obtain information as much as possible, depends onnot only the progress of analysis methods, but also the improvement of signal acquisition and the condi-tions for further analysis. Without automatic data processing, the analysis of experimental data might bewith poor accuracy,time-consuming and even impossible in some new cardiov…     

Novel Approach for Endothelializing Vascular Devices: Understanding and Exploiting Elastin–Endothelial Interactions

Elastin is an essential component of arteries which provides structural integrity and instructs smooth muscle cells to adopt a quiescent state. Despite interaction of endothelial cells with elastin in the internal elastic lamina, the potential for exploiting this interaction therapeutically has not been explored in detail. In this study, we show that tropoelastin (a precursor of elastin) stimulates endothelial cell migration and adhesion more than smooth muscle cells. The biological activity of tropoelastin on endothelial cells is contained in the VGVAPG domain and in the carboxy-terminal 17-amino acids. We show that the effects of the carboxy-terminal 17 amino acids, but not those of VGVAPG, are mediated by integrin α_Vβ₃. We demonstrate that tropoelastin covalently linked to stainless steel disks promotes adhesion of endothelial progenitor cells and endothelial cells to the metal surfaces. The adherent cells on the tropoelastin-coated metal surfaces form monolayers that can withstand and respond to arterial shear stress. Because of the unique effects of tropoelastin on endothelial and smooth muscle cells, coating intravascular devices with tropoelastin may stimulate their endothelialization, inhibit smooth muscle hyperplasia, and improve device performance.     

Tools for Detection of Mycoplasma amphoriforme: a Primary Respiratory Pathogen?

Mycoplasma amphoriforme is a recently described organism isolated from the respiratory tracts of patients with immunodeficiency and evidence of chronic infection. Novel assays for the molecular detection of the organism by real-time quantitative PCRs (qPCRs) targeting the uracil DNA glycosylase gene (udg) or the 23S rRNA gene are described here. The analytical sensitivities are similar to the existing conventional M. amphoriforme 16S rRNA gene PCR, with the advantage of being species specific, rapid, and quantitative. By using these techniques, we demonstrate the presence of this organism in 17 (19.3%) primary antibody-deficient (PAD) patients, 4 (5%) adults with lower respiratory tract infection, 1 (2.6%) sputum sample from a patient attending a chest clinic, and 23 (0.21%) samples submitted for viral diagnosis of respiratory infection, but not in normal adult control subjects. These data show the presence of this microorganism in respiratory patients and suggest that M. amphoriforme may infect both immunocompetent and immunocompromised people. Further studies to characterize this organism are required, and this report provides the tools that may be used by other research groups to investigate its pathogenic potential.     

Immuno‐Quantitative Polymerase Chain Reaction for Detection and Quantitation of Prion Protein

Abstract

Immuno‐polymerase chain reaction (PCR) is an extremely sensitive detection method, combining the specificity of antibody detection and the sensitivity of PCR. We have developed an immuno‐quantitative PCR (iqPCR), exploiting real‐time PCR technology, in order to improve this immuno‐detection method and make it quantitative. To illustrate the advantages of iqPCR, we have compared it with a conventional enzyme linked immuno sorbent assay (ELISA) technique in experiments aimed at detecting the cellular and the resistant form of prion protein in bovine brain extract. The iqPCR technique proved to be more sensitive than ELISA, so it could be a technique of choice for the diagnosis of infected animals both at an ante mortem and post‐mortem stage.     

Notes: Can We Use Imipenem and Meropenem Vitek 2 MICs for Detection of Suspected KPC and Other-Carbapenemase Producers among Species of Enterobacteriaceae?

Imipenem and meropenem Vitek 2 MICs were evaluated for a panel of 104 Enterobacteriaceae for identification of carbapenemase producers. The sensitivity and specificity values for the new CLSI interpretative criteria (CLSI document M100-S20-U, 2010) were 98% and 83% for imipenem and 76% and 83% for meropenem, respectively. We propose an algorithm that is highly sensitive (98%) and specific (94%) for carbapenemase screening based on the combined use of imipenem and meropenem MICs.     

Detection of deletions in de novo “balanced” chromosome rearrangements: Further evidence for their role in phenotypic abnormalities

Purpose: The purpose of this study was to test the hypothesis that deletions of varying sizes in de novo apparently balanced chromosome rearrangements are a significant cause of phenotypic abnormalities.Methods: A total of fifteen patients, with seemingly balanced de novo rearrangements by routine cytogenetic analysis but with phenotypic anomalies, were systematically analyzed. We characterized the breakpoints in these fifteen cases (two of which were ascertained prenatally), using a combination of high-resolution GTG-banding, fluorescence in situ hybridization (FISH) with bacterial artificial chromosomes (BACs), and data from the Human Genome Project.Results: Molecular cytogenetic characterization of the 15 patients revealed nine with deletions, ranging in size from 0.8 to 15.3 Mb, with the number of genes lost ranging from 15 to 70. In five of the other six cases, a known or putative gene(s) was potentially disrupted as a result of the chromosomal rearrangement. In the remaining case, no deletions were detected, and no known genes were apparently disrupted.Conclusions: Our study suggests that the use of molecular cytogenetic techniques is a highly effective way of systematically delineating chromosomal breakpoints, and that the presence of deletions of varying size is an important cause of phenotypic abnormalities in patients with “balanced” de novo rearrangements.     

Time�CFrequency Approaches for the Detection of Interactions and Temporal Properties in Renal Autoregulation

We compare the influence of time?Cfrequency methods on analysis of time-varying renal autoregulation properties. Particularly, we examine if detection probabilities are similar for amplitude and frequency modulation for a modulated simulation signal among five time?Cfrequency approaches, and if time-varying changes in system gain are detected using four approaches for estimating time-varying transfer functions. Detection of amplitude and frequency modulation varied among methods and was dependent upon background noise added to the simulated data. Three non-parametric time?Cfrequency methods accurately detected modulation at low frequencies across noise levels but not high frequencies; while the converse was true for a fourth, and a fifth non-parametric approach was not capable of modulation detection. When applied to estimation of time-varying transfer functions, the parametric approach provided the most accurate estimations of system gain changes, detecting a 1?dB step increase. Application of the appropriate methods to laser Doppler recordings of cortical blood flow and arterial pressure data in anesthetized rats reaffirm the presence of time-varying dynamics in renal autoregulation. An increase in the peak system gain and detection of amplitude modulation of the Myogenic mechanism both occurred after inhibition of nitric oxide synthase, suggesting a connection between the operation of underlying regulators and system performance.     

Russian “Successful” Clone B0/W148 of Mycobacterium tuberculosis Beijing Genotype: a Multiplex PCR Assay for Rapid Detection and Global Screening

We describe a multiplex PCR assay to detect the Mycobacterium tuberculosis Beijing genotype variant B0/W148, which is considered a “successful” clone of M. tuberculosis, widespread in Russia. Specificity and sensitivity of the assay were 100% based on the analysis of a collection of 516 M. tuberculosis isolates of different genotypes and origins. This assay may be used for accurate and simple detection and surveillance of this clinically and epidemiologically important variant of M. tuberculosis.     

IgG,IgA and IgM Antibodies against FSH: Serological Markers of Pathogenic Autoimmunity or of Normal Immunoregulation?

PROBLEM: Autoimmune mechanisms are often involved in causing infertility. Among the possible targets of autoantibodies, the follicle-stimulating hormone (FSH) which regulates the follicular maturation in human ovary is a promising candidate. We aimed to study whether anti-FSH-antibodies might be involved in different clinical types of infertility. METHOD OF STUDY: The study group consisted of 178 patients (75 with polycystic ovary syndrome (PCOS), 103 with endometriosis) and 75 pregnant women. Female blood donors formed the control group (n = 85). Indirect enzyme-linked immunosorbent assay tests were performed using purified FSH as antigens and a synthetic peptide corresponding to the 78-93 region (V14D) of the human FSH beta-chain. CONCLUSION: We showed that anti-FSH-antibodies were present in controls and their production decreased during pregnancy. Endometriosis and PCOS were associated with higher values of anti-FSH-immunoglobulin (Ig)A, anti-V14D-IgA, and endometriosis with anti-V14D-IgG. Our data suggest that anti-FSH-IgA could be a marker of ovarian disorders that cause infertility.     

Detection of “Candidatus Neoehrlichia mikurensis” in Two Patients with Severe Febrile Illnesses: Evidence for a European Sequence Variant

Recently, a new genus of Anaplasmataceae termed “Candidatus Neoehrlichia” was discovered in ticks and rodents. Here, we report on two patients who suffered from febrile bacteremia due to “Candidatus Neoehrlichia mikurensis” associated with thrombotic or hemorrhagic events. 16S rRNA and groEL gene sequencing provided evidence of three groups of sequence variants.     

Detection of an “epimastigote-like” intracellular stage ofTrypanosoma cruzi

Monoclonal antibody (mAb) DION 5.1b, derived from mice immunized withTrypanosoma dionisii, recognizes a 72/76-kD surface glycoprotein specific to the epimastigote stage ofT. dionisii andT. cruzi. None of the three other stages of theT. cruzi life cycle expresses any DION 5.1b-specific epitope. However, mAb DION 5.1b labels an intracellular form with epimastigote-like morphology that appears to be late and transient in the intracellular cycle. This result suggests that the morphological similarity between the observed epimastigote-like intracellular form in mammals and the epimastigote form in insects may extend to the antigenic pattern.     

Effects of Denaturation Agents on Antigenic Expression and Levels of Nonspecific Background Staining for 5-Bromo-2′-Deoxyuridine Immunohistochemistry

Abstract

The capillary gap procedure by Frank et a1 in 1993 used to intensify irnmunohistochemical demonstration of the thymidine analog, 5-bromo-2'deoxyuridine (BrdU) has proved useful for quantitation of chemically induced cell proliferation. Localization of BrdU in fixed tissue requires that DNA be denatured to single strands with acids, bases, heat, or fixatives. To determine if staining intensity and levels of background staining were affected by the method of denaturation, the following denaturation agents were compared: 4 M and 2 M hydrochloric acid for 20 min at room temperature; Bouin fixative for 60 min or 30 min at 60°C; deionized water (DI) heated in a microwave on high for 10 rnin (approximately 96–98°C) followed by 10 min of cooling; and DI water at room temperature. Slide 1 of each pair consisted of serial sections of formalin fixed rat kidney and duodenum. Slide 2 of each pair consisted of serial sections of both Bouin fixed rat testes and formalin fixed rat duodenum. Four M hydrochloric acid and Bouin at 60°C for 1 hr gave the most intense nuclear expression for formalin fixed tissue. Following DI water at room temperature, excellent BrdU staining occurred for Bouin fixed testes; however, formalin fixedtissues wereminimally labeled and showed nonspecific background staining. Microwave heating resulted in tissue destruction or increased background. Bouin fixative at 60°C for 1 hr gave the best BrdU staining with little or no nonspecific background staining in any tissue evaluated. (The J Histotechnol 18:301, 1995)     

Collective Cell Migration: “All for One and One for All”

Cell migration is a key mechanism during neural development, as it allows cells to reach their final destination from their birthplace. In some cases, cells migrate in isolation, whereas in others they migrate in collectives, as chains, streams, clusters, or sheets. The coordinated and timely process of collective migration eventually ensures the proper organization of the nervous system and its misregulation leads to severe diseases, including neurological disorders. This review impinges upon the cellular and molecular interactions underlying collective cell migration in animal models, and highlights the recent advances made through in vivo analyses of the Drosophila wing glia.