A Strategy for Sensitive,Large Scale Quantitative Metabolomics

Metabolite profiling has been a valuable asset in the study of metabolism in health and disease. However, current platforms have different limiting factors, such as labor intensive sample preparations, low detection limits, slow scan speeds, intensive method optimization for each metabolite, and the inability to measure both positively and negatively charged ions in single experiments. Therefore, a novel metabolomics protocol could advance metabolomics studies. Amide-based hydrophilic chromatography enables polar metabolite analysis without any chemical derivatization. High resolution MS using the Q-Exactive (QE-MS) has improved ion optics, increased scan speeds (256 msec at resolution 70,000), and has the capability of carrying out positive/negative switching. Using a cold methanol extraction strategy, and coupling an amide column with QE-MS enables robust detection of 168 targeted polar metabolites and thousands of additional features simultaneously.  Data processing is carried out with commercially available software in a highly efficient way, and unknown features extracted from the mass spectra can be queried in databases.     

Location of Calcium Within Bacillus Spores by Electron Probe X-Ray Microanalysis

Spectroscopic microanalysis of the element-characteristic X rays produced by a scanning electron microprobe was employed to detect calcium and carbon in both intact and thin-sectioned spores of Bacillus cereus T and B. megaterium QM B1551. Linear scan profiles and multilinear scan images of the X-ray emissions for calcium (Ca(Kalpha)) were compared with those for carbon (C(Kalpha)) as an index of mass. Location was accomplished by stereological comparisons with secondary electron images and conventional transmission electron micrographs. Although the elements could be detected at the attogram level theoretically, spatial resolution was limited to approximately 500 to 1,000 nm in an intact spore, e.g., by the primary electron beam diameter, the electron-excited spore microvolume, and the type of specimen support. The resolution was improved to approximately 100 to 200 nm by use of thin-sectioned spores, with precautions to prevent calcium leakage from the specimen during preparations. In both intact and sectioned spores, calcium was distributed throughout the spore, similarly to carbon, and concentrated mainly in a central region corresponding to the spore protoplast.     

TV Time but Not Computer Time Is Associated with Cardiometabolic Risk in Dutch Young Adults

Background

TV time and total sedentary time have been positively related to biomarkers of cardiometabolic risk in adults. We aim to examine the association of TV time and computer time separately with cardiometabolic biomarkers in young adults. Additionally, the mediating role of waist circumference (WC) is studied.

Methods and Findings

Data of 634 Dutch young adults (18–28 years; 39% male) were used. Cardiometabolic biomarkers included indicators of overweight, blood pressure, blood levels of fasting plasma insulin, cholesterol, glucose, triglycerides and a clustered cardiometabolic risk score. Linear regression analyses were used to assess the cross-sectional association of self-reported TV and computer time with cardiometabolic biomarkers, adjusting for demographic and lifestyle factors. Mediation by WC was checked using the product-of-coefficient method.TV time was significantly associated with triglycerides (B = 0.004; CI = [0.001;0.05]) and insulin (B = 0.10; CI = [0.01;0.20]). Computer time was not significantly associated with any of the cardiometabolic biomarkers. We found no evidence for WC to mediate the association of TV time or computer time with cardiometabolic biomarkers.

Conclusions

We found a significantly positive association of TV time with cardiometabolic biomarkers. In addition, we found no evidence for WC as a mediator of this association. Our findings suggest a need to distinguish between TV time and computer time within future guidelines for screen time.     

Reversed Halo Sign: Presents in Different Pulmonary Diseases

Objective

To observe the incidence of reversed halo sign in different pulmonary diseases and the pathological correspondence of reversed halo sign.

Methods

Retrospectively studied the high resolution computer tomography scans of all the patients who were admitted in our department with abnormal pulmonary imaging, from 1st of January 2011 to 31st of December 2013, and all the cases with reversed halo sign on the high resolution computer tomography were collected. Clinical data such as pathological findings and confirmed diagnosis of the patients with reversed halo sign on the high resolution computer tomography scan were collected and summarized.

Results

Of 1546 abnormal High resolution computer tomography scans 108 had a reverse halo sign present, including 108 cases were observed with reversed halo sign in the high resolution computer tomography, including 40 cases of pulmonary tuberculosis, 43 cases of cryptogenic organizing pneumonia, 16 cases of lung cancer, 7 cases of sarcoidosis, and 1 case of pulmonary cryptococcosis, 1 case of granulomatosis with polyangiitis. Reversed halo sign had a higher incidence in granulomatous diseases (16.28%) compared with non-granulomatous diseases (9.97%).

Conclusions

Reversed halo sign is relatively non specific; it can be observed in different lung diseases, and different phases of diseases; reversed halo sign is more commonly found in granulomatous diseases compared with non-granulomatous diseases, and is most commonly observed in pulmonary tuberculosis among the granulomatous diseases, and in cryptogenic organizing pneumonia among the non-granulomatous diseases.     

High-resolution cytometry of FISH dots in interphase cell nuclei.

BACKGROUND: Flow cytometry (FCM) and laser scanning cytometry (LSCM) provide indispensable tools for measuring large number of cells with low resolution. Confocal microscopy, on the other hand, is used for measuring small number of cells with high resolution. In this paper, we present a reasonable compromise between the two extremes. METHODS: We have developed a completely automated, high-resolution system (high-resolution cytometer, HRCM) capable of analyzing microscope slides with FISH-stained interphase nuclei in two dimensions as well as in three dimensions using a fully motorized epi-fluorescence microscope and a cooled digital CCD camera fully controlled by a high-performance computer which performs both acquisition and related on-line image analysis. The images of different dyes are acquired sequentially using highly specific filters and superimposed in computer memory. For each nucleus and each hybridization dot, user-selected attributes (such as position, size, intensity, etc.) are computed off-line using another processor or computer connected with a network. RESULTS: Using HRCM, it is possible to analyze multi-color preparations including UV-excited dyes as well as repeatedly hybridized preparations reacquiring individual nuclei. The speed of the acquisition and analysis is about 50 nuclei per minute in two dimensions and 1 nucleus per minute in three dimensions, but depends on the density of nuclei on the slide; the precision of the lateral and axial measurements is approximately 100 nm. CONCLUSIONS: Thus, using overnight acquisition, quantities comparable to those of FCM or LSCM measurements can be analyzed with an accuracy comparable to confocal microscopy. HRCM is suitable for a number of clinical and scientific tasks: routine diagnostics, follow-up of therapy, studies of chromatin structure, and many other different aspects of cell research.     

Association of Family Environment with Children's Television Viewing and with Low Level of Physical Activity

Objective: This study examined associations between the family environment and children's television (TV) viewing and likelihood of being low‐active. Research Methods and Procedures: In 2001, children were recruited from 19 primary schools in Melbourne, Australia. Parents completed a questionnaire about their child's TV viewing and the family environment. Children also completed a questionnaire and wore an accelerometer for 8 days. Movement counts were used to identify low‐active children (lowest quartile). Data were analyzed in May 2004. Results: The sample consisted of 878 children (mean age = 11.5 ± 0.6 yrs). Multiple logistic regression revealed that socioeconomic status [adjusted odds ratios (AOR) = 0.4 boys], frequency families watched TV together (AOR = 2.0 boys), mothers’ (AOR = 1.8 boys; AOR = 2.5 girls) and fathers’ (AOR = 2.6 boys; AOR = 2.8 girls) TV viewing, and rules prohibiting TV during mealtimes (AOR = 0.6 boys; AOR = 0.6 girls) related to children watching TV ≥2 h/d. Variables associated with low‐level physical activity included self‐reported enjoyment of Internet use (AOR = 1.7 boys) and preference for watching TV (AOR = 2.3 girls), perception that mother uses computer a lot (AOR = 1.9 boys) and likes using the computer (AOR = 0.6 girls), fathers’ reported computer/electronic games use (AOR = 1.7 girls), frequency families used computer together (AOR = 0.4 girls), rules that TV viewing must be supervised (AOR = 1.9 boys; AOR = 0.6 girls), and having pay TV (AOR = 0.6 boys) and electronic games at home (AOR = 2.6 boys). Discussion: These findings suggest that the relationships between the family environment and TV viewing and low‐level activity are complex and that these behaviors are distinct.     

A selective mapping algorithm for computer analysis of voided urine cell images

One of the fundamental targets of the automated image analysis of cytologic preparations is the reduction of computer classification errors due to cells or other objects that do not lend themselves to image segmentation or that have morphologic features that may mislead the cell classification schemes. In prior work from this laboratory, the achievement of this goal was attempted by hierarchical analysis of sequential microscopic objects at high resolution. This paper reports on the successful development and implementation of an automated "selective mapping algorithm" that selects cells at low power for further analysis and eliminates a large proportion of unwanted "objects." The algorithm classifies the objects and extracts appropriate features from a 256 X 240 digital image obtained via a 10 X planachromatic objective. The five-node binary tree classifier used in this triage is described. The algorithm was trained and tested initially on 501 visually classified microscopic "objects," resulting in a correct acceptance rate of 61.3% and correct rejection rate of 81.3%. The selective mapping algorithm was subsequently integrated into the video-based image analysis system constructed at the Montefiore Medical Center for the diagnostic evaluation of sediments of voided urine. The algorithm was then tested on ten cytocentrifuge preparations for a preliminary evaluation of its performance. Up to 100 "objects" per case were selected by the algorithm for further classification by the computer at high power. Of the 810 "objects" selected by the selective mapping algorithm, 344 (42.5%) were classified by the computer at high resolution as cells of diagnostic value ("WELL" cells) and 466 were rejected.(ABSTRACT TRUNCATED AT 250 WORDS)     

Time-lapse imaging of dendritic spines in vitro

Dendritic spines are small protrusions present postsynaptically at approximately 90% of excitatory synapses in the brain. Spines undergo rapid spontaneous changes in shape that are thought to be important for alterations in synaptic connectivity underlying learning and memory. Visualization of these dynamic changes in spine morphology are especially challenging because of the small size of spines (approximately 1 microm). Here we describe a microscope system, based on a spinning-disk confocal microscope, suitable for imaging mature dendritic spines in brain slice preparations, with a time resolution of seconds. We discuss two commonly used in vitro brain slice preparations and methods for transfecting them. Preparation and transfection require approximately 1 d, after which slices must be cultured for at least 21 d to obtain spines of mature morphology. We also describe imaging and computer analysis routines for studying spine motility. These procedures require in the order of 2 to 4 h.     

Physical characterisation of plasmids in a morpholine-degrading mycobacterium

The plasmid content of an environmental mycobacterium (MorG) which degrades morpholine (Mor+ phenotype) was investigated. The combination, in what appears to be a novel way, of restriction endonuclease digestion, gel electrophoresis and scanning densitometry permitted the resolution of mixed plasmid preparations into four distinct plasmids with sizes of 54 (approximately), 27.7, 22.8 and 22.6 kb. These plasmids were named pMOR1, 2, 3 and 4, respectively. The Mor+ phenotype was found to be unstable during acriflavin treatment. In four independently isolated Mor- mutants, plasmid pMOR2 was found to have acquired an insert of approximately 1.8 kb within a specific 5.9 kb BamHI fragment. It is concluded that pMOR2 is involved in the coding of the Mor+ phenotype.     

Bacillus thuringiensis and neem seed oil (Azadirachta indica) effects on the potato tuber moth Phthorimaea operculella zeller in the field and stores

A comparative evaluation for the efficacy of Bacillus thuringiensis and neem seed oil on Phthorimaea operculella has been carried out in the field and store. These two preparations were almost equally effective on the potato tuber moth infestation. The percentage of infestation was reduced through successive application of either preparations in the field up to harvest. No synergism was observed upon using combination of the two preparations. In the store, neem seed oil (500 ppm) was highly protective and was as effective as sevin. A combination of both neem and B.t. (Delfin) significantly protects the tubers. This suggests the possible use of either neem seed oil or B.t. in combating the insect pest in the field or during storage.     

High-speed, random-access fluorescence microscopy: I. High-resolution optical recording with voltage-sensitive dyes and ion indicators.

The design and implementation of a high-speed, random-access, laser-scanning fluorescence microscope configured to record fast physiological signals from small neuronal structures with high spatiotemporal resolution is presented. The laser-scanning capability of this nonimaging microscope is provided by two orthogonal acousto-optic deflectors under computer control. Each scanning point can be randomly accessed and has a positioning time of 3-5 microseconds. Sampling time is also computer-controlled and can be varied to maximize the signal-to-noise ratio. Acquisition rates up to 200k samples/s at 16-bit digitizing resolution are possible. The spatial resolution of this instrument is determined by the minimal spot size at the level of the preparation (i.e., 2-7 microns). Scanning points are selected interactively from a reference image collected with differential interference contrast optics and a video camera. Frame rates up to 5 kHz are easily attainable. Intrinsic variations in laser light intensity and scanning spot brightness are overcome by an on-line signal-processing scheme. Representative records obtained with this instrument by using voltage-sensitive dyes and calcium indicators demonstrate the ability to make fast, high-fidelity measurements of membrane potential and intracellular calcium at high spatial resolution (2 microns) without any temporal averaging.     

Detection of Signature Sequences in Overlapping Genes and Prediction of a Novel Overlapping Gene in Hepatitis G Virus

In viruses an increased coding ability is provided by overlapping genes, in which two alternative open reading frames (ORFs) may be translated to yield two distinct proteins. The identification of signature sequences in overlapping genes is a topic of particular interest, since additional out-of-frame coding regions can be nested within known genes. In this work, a novel feature peculiar to overlapping coding regions is presented. It was detected by analysis of a sample set of 21 virus genomic sequences and consisted in the repeated occurrence of a cluster of basic amino acid residues, encoded by a frame, combined to a stretch of acidic residues, encoded by the corresponding overlapping frame. A computer scan of an additional set of virus sequences demonstrated that this feature is common to several other known overlapping ORFs and led to prediction of a novel overlapping gene in hepatitis G virus (HGV). The occurrence of a bifunctional coding region in HGV was also supported by its extremely lower rate of synonymous nucleotide substitutions compared to that observed in the other gene regions of the HGV genome. Analysis of the amino acid sequence that was deduced from the putative overlapping gene revealed a high content of basic residues and the presence of a nuclear targeting signal; these characteristics suggest that a core-like protein may be expressed by this novel ORF. Received: 21 July 1999 / Accepted: 26 October 1999     

Translational bypassing without peptidyl-tRNA anticodon scanning of coding gap mRNA

Half the ribosomes translating the mRNA for phage T4 gene 60 topoisomerase subunit bypass a 50 nucleotide coding gap between codons 46 and 47. The pairing of codon 46 with its cognate peptidyl-tRNA anticodon dissociates, and following mRNA slippage, peptidyl-tRNA re-pairs to mRNA at a matched triplet 5' adjacent to codon 47, where translation resumes. Here, in studies with gene 60 cassettes, it is shown that the peptidyl-tRNA anticodon does not scan the intervening sequence for potential complementarity. However, certain coding gap mutants allow peptidyl-tRNA to scan sequences in the bypassed segment. A model is proposed in which the coding gap mRNA enters the ribosomal A-site and forms a structure that precludes peptidyl-tRNA scanning of its sequence. Dissipation of this RNA structure, together with the contribution of 16S rRNA anti-Shine-Dalgarno sequence pairing with GAG, facilitates peptidyl-tRNA re-pairing to mRNA.     

Phylogeny of Dicranophoridae (Rotifera: Monogononta) – a maximum parsimony analysis based on morphological characters

This study presents the first phylogenetic analysis of Dicranophoridae (Rotifera: Monogononta), a species rich rotifer family of about 230 species currently recognized. It is based on a maximum parsimony analysis including 77 selected ingroup and three outgroup taxa and a total of 59 phylogenetically informative morphological characters. Character coding is based on personal investigation of material collected by the authors and an extensive survey of the literature. Apart from covering general body organization, character coding primarily relies on scanning electron microscopic preparations of the mastax jaw elements. Our study suggests monophyly of Dicranophoridae with a clade of Dicranophorus and Dorria as the sister taxon of all other dicranophorid species. Monophyly of Encentrum , the most species rich genus within Dicranophoridae, cannot be demonstrated. Within Dicranophoridae our study identifies the monophyletic taxa Caudosubbasifenestrata, Intramalleata, Praeuncinata and Proventriculata, each based on unambiguous character transformations evolved in their stem lineages. However, resolution within Praeuncinata and Proventriculata is very limited. Although some terminal clades within Praeuncinata and Proventriculata are recognized, basal splits remain obscure. Probably, other characters such as DNA sequence data are needed to further our understanding of phylogenetic relationships within these poorly resolved taxa.     

Microfluorimetric scanning of sympathetic nerve fibers. An improved method to quantitate formaldehyde induced fluorescence of biogenic amines

A two dimensional scanning procedure was developed for studying quantitative aspects of formaldehyde induced fluorescence (FIF) from noradrenaline containing nerve fibers. A computer (PDP 12) controlled scanning device was equipped with a cooled photomultiplier, a 0.5 micron scanning stage and a 0.5 micron measuring spot. Photodecomposition was reduced by a high scanspeed (100/sec) and a small excitation field. Digitized images were obtained with a high resolution. Data (10,000/scan) of large scan areas (2500 micron2) in the iris of the rat were transformed into fluorescence histograms, which allowed statistical evaluation. Visible changes induced in the histochemical preparation by administration of drugs (reserpine, L-Dopa) were reflected in characteristic changes in the histograms. A minor reduction in noradrenaline content, which was too small to be detected by visual observation of the microscopical image and by classical microfluorimetric methods, did evoke a significant change in the histograms. It is concluded that microfluorimetric scanning is a very sensitive technique for detecting changes of formaldehyde induced fluorescence from neuronal networks.     

TV viewing, computer game playing and nightmares in school children.

Often the question arises whether TV viewing or playing computer games, especially those of violent and aggressive content, has negative effects on children's sleep patterns and dreaming. Empirical data on this topic, however, are scarce. Two hundred fifty-two schoolchildren (range: 9 to 13 years) completed a questionnaire about media use and nightmares. The findings indicates that interindividual differences in nightmare frequency were not explained by interindividual differences in TV viewing or computer game playing habits. In addition, no effect of the TV films the evening before completing the questionnaire on the dreams the subsequent night could be demonstrated. Since the approach could not rule out an effect of media use habits on nightmares, the next step will be a longitudinal study eliciting the media use habits and the occurrence of nightmares by applying self-rated scales as well as information obtained by parents. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

单排CT轴位螺旋扫描联合冠状面重组技术对鼻腔鼻窦检查的意义

目的：研究在单排CT轴位螺旋扫描联合多层面重组技术重建冠状面影像诊断鼻腔疾病,评价其诊断价值和可行性。方法：对2012年1月-2013年3月我院耳鼻咽喉科进行单排CT轴位螺旋扫描的患者,共219例,对这些患者加做冠状位扫描,比较重建后的影像资料与直接冠状位扫描的图像的分辨率情况,并评估重建质量及运用价值。,结果：单排CT轴位螺旋扫描后冠状面重组的图像：边界清楚、完整,数据无丢失。利用重组技术后重建的结构清晰、连续,能够真实地反映鼻腔、鼻窦及病变结构的实际情况。但分辨率较直接冠状位平扫的患者稍差。结论：单排CT轴位螺旋扫描后冠状面重组的图像,虽然分辨率不如直接冠状位扫描图像高,但与解剖结构、原图像等相结合仍能分辨出病变情况,基本可以满足诊断要求和临床需要。     

Characterization of the terminal cisternae and longitudinal tubules of sarcoplasmic reticulum from malignant hyperpyrexia susceptible porcine skeletal muscle

1. The sarcoplasmic reticulum (SR) from malignant hyperpyrexia susceptible (MHS) and control porcine skeletal muscle was separated into vesicular fractions enriched in the membrane elements of the terminal cisternae and longitudinal tubules. 2. The two membrane preparations were highly purified and had distinctive features which were associated with their origins in the SR membraneous network. 3. Calsequestrin and calcium were enriched in the terminal cisternae fraction (HSR), in comparison to longitudinal tubule preparations (LSR). 4. The HSR membrane also had a greater total capacity to store Ca2+ and Ca2+ release was more rapid than from LSR preparations. 5. No distinction could be made between the membrane morphology, Ca2+ -fluxes or Ca2+ -dependent ATPase activities, associated with these functionally distinct regions of MHS and control preparations.     

Media Use and Dreaming: The Relationship Among Television Viewing, Computer Game Play, and Nightmares or Pleasant Dreams.

Children (N=2,546) in a random sample of 15 secondary schools in Flanders, Belgium, completed a questionnaire about volume of television viewing, computer game playing, and nightmares and pleasant dreams related to these activities. TV content showed up frequently in nightmares for 33% of the children, and computer games were associated with nightmares in about 10% of boys and 5% of girls. About 60% of 13-year-olds and 50% of 16-year-olds reported having pleasant dreams related to TV. Computer games also appeared in pleasant dreams for a majority of 13-year-olds and a minority of 16-year-olds. Media influence on dream content was not limited to excessive media users. Concerns about the implications for the impact of media on dreams and general psychological well-being are discussed. (PsycINFO Database Record (c) 2010 APA, all rights reserved)     

Effect of microcomputed tomography voxel size on the finite element model accuracy for human cancellous bone

The level of structural detail that can be acquired and incorporated in a finite element (FE) analysis might greatly influence the results of microcomputed tomography (microCT)-based FE simulations, especially when relatively large bones, such as whole vertebrae, are of concern. We evaluated the effect of scanning and reconstruction voxel size on the microCT-based FE analyses of human cancellous tissue samples for fixed- and free-end boundary conditions using different combinations of scan/reconstruction voxel size. We found that the bone volume fraction (BV/TV) did not differ considerably between images scanned at 21 and 50 microm and reconstructed at 21, 50, or 110 microm (-0.5% to 7.8% change from the 21/21 microm case). For the images scanned and reconstructed at 110 microm, however, there was a large increase in BV/TV compared to the 21/21 microm case (58.7%). Fixed-end boundary conditions resulted in 1.8% [coefficient of variation (COV)] to 14.6% (E) difference from the free-end case. Dependence of model output parameters on scanning and reconstruction voxel size was similar between free- and fixed-end simulations. Up to 26%, 30%, 17.8%, and 32.3% difference in modulus (E), and average (VMExp), standard deviation (VMSD) and coefficient of variation (COV) of von Mises stresses, respectively, was observed between the 21/21 microm case and other scan/reconstruction combinations within the same (free or fixed) simulation group. Observed differences were largely attributable to scanning resolution, although reconstruction resolution also contributed significantly at the largest voxel sizes. All 21/21 microm results (taken as the gold standard) could be predicted from the 21/50 (r2adj= 0.91-0.99;p<0.001), 21/110 (r2adj =0.58-0.99;p<0.02) and 50/50 results (r2adj=0.61-0.97;p<0.02). While BV/TV, VMSD, and VMExp/sigma(z) from the 21/21 could be predicted by those from the 50/110 (r2adj =0.63-0.93;p<0.02) and 110/110 (r2adj =0.41-0.77;p<0.05) simulations as well, prediction of E, VMExp, and COV became marginally significant (0.04相似文献