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PubMed1991

Isolation and characterization of a cytosolic aldehyde dehydrogenase-encoding cDNA from mouse liver.

A series of overlapping cDNA clones encoding AHD-2, the constitutive liver cytosolic aldehyde dehydrogenase of the mouse, were isolated from lambda gt11 libraries by screening with antibody and hybridization probes. Together, these clones span 2036 bp, virtually the entire Ahd-2 transcript. The protein corresponding to the cDNA contains 501 amino acids (aa) with a predicted Mr of 54467. The aa sequence is 87% and 84% identical to liver cytosolic AHDs of human and horse, respectively. Major and minor transcription start points (tsp), located 43 and 44 bp upstream from the ATG codon, respectively, were identified by primer extension analysis. The sequence of a 700-bp genomic fragment containing the first exon and 546 bp of 5'-flanking DNA was determined. TATA and CCAAT motifs are located 31 and 73 bp, respectively, from the major tsp.

Aldehyde Dehydrogenase↗

PubMed2006

Seeking completeness in bacterial mutant hunts.

The identification of most or all of the genetic functions that are required for a particular biological process could be achieved through phenotypic studies of high genome-coverage mutant collections. Technologies for creating such collections, in the form of mixed populations or individually arrayed sequence-defined mutants, are now available for numerous bacterial species. The analysis of mixed mutant collections using microarray-based detection procedures appears to be particularly effective in identifying functions required for complex processes such as virulence. The phenotypic analysis of sequence-defined mutant libraries provides a virtually complete identification of nonessential genes required for processes for which suitable screens can be devised. Such libraries also serve as a source of individual mutants for examining the biological relevance of gene associations revealed by transcriptional profiling or homology.

Bacteria↗

PubMed2002

Manufacture and composition of fresh frozen plasma and virus-inactivated therapeutic plasma preparations: correlation between composition and therapeutic efficacy.

The clinical efficacy of the therapeutic plasma used in the treatment of congenital and acquired severe coagulopathy depends on the potency of clotting factor and inhibitor activities. The composition of plasma strongly depends on the conditions under which it is produced. A low citrate anticoagulant-to-blood ratio, short intervals between donation and plasma separation and rapid freezing markedly improve the preservation of unstable coagulation factors. The influence of different leukocyte reduction filters on plasma quality still requires clarification. Recent trials on long-term storage conditions suggest that keeping plasma at -30 degrees C or colder over a period of 24-36 months prevents substantial decrease in clotting factor activities including factor VIII (FVIII). Three types of therapeutic plasma are currently available. Quarantine-stored fresh frozen plasma (FFP) contains physiological activities of therapeutically relevant plasma proteins, but carries a risk of transmitting blood-borne viruses that cannot be detected by human immunodeficiency virus (HIV) and hepatitis B and C screening. In contrast, solvent/detergent-treated plasma (SDP) and methylene blue/light-treated plasma (MBP) is virtually free of HIV and hepatitis C virus (HCV) subtypes. Virus inactivation procedures can have the consequence of reducing several clotting factors and inhibitors in SDP and MBP to varying degrees. However, pooling of plasma units before solvent/detergent (SD) treatment results in well-standardized protein levels of SDP. At least five prospective trials and four observational studies covering different clinical settings suggest that SDP and FFP do not substantially differ in their clinical efficacy or in their tolerance. By way of contrast, there is a lack of data about the clinical efficacy and tolerance of MBP compared to FFP.

Blood Coagulation Factors↗

PubMed2000

Myocardial perfusion scintigraphy as a screening method for significant coronary artery stenosis in cardiac transplant recipients.

BACKGROUND: Several studies have explored the feasibility of using myocardial perfusion imaging to detect allograft vasculopathy after heart transplantation. We undertook the present prospective consecutive study to comparatively evaluate the role of serial myocardial perfusion single-photon emission computed tomography (SPECT) scanning and coronary arteriography (CAG) in detecting coronary artery stenosis suitable for coronary angioplasty in heart transplant recipients. METHODS: Within a 2-week interval during a follow-up period of 5.6 (95% confidence limits 2.1 to 12) years, 255 serial CAGs and myocardial perfusion scintigraphies were performed in 67 patients. Arteriography and scintigraphy were performed once yearly after heart transplantation. We retrospectively analyzed the data. RESULTS: Myocardial scintigraphy showed pathologic reversible defects in 9 out of 67 patients. Four of these patients had significant (>50% and also >70%) focal segmental stenosis in the middle and proximal parts of the coronary arteries (Type A lesions), 1 had diffuse and circumferential narrowing in the distal parts (Type B lesions), whereas CAG showed no lesions in the remaining 4 patients. The patients with significant Type A lesions were revascularized with percutaneous coronary angioplasty. Coronary arteriography showed that 1 patient had extensive Type A and Type B lesions, whereas myocardial perfusion scans detected no. The predictive value of a negative (normal) SPECT was 98% (95% confidence limits 94% to 100%) for the detection of lesions suited for revascularization. CONCLUSIONS: Annual myocardial SPECT seems well suited to screen for significant coronary artery stenosis. A SPECT study without reversible defects virtually excludes lesions suitable for coronary artery revascularization.

Adolescent↗

PubMed1982

Rigid analogues of dopamine: synthesis and interaction of 6-exo- and 6-endo-(3',4'-dihydroxyphenyl)-2-azabicyclo[2.2.2]octanes with dopamine uptake sites and receptors.

Two isomeric 6-endo- and 6-exo-(3',4'-dihydroxyphenyl) derivatives (1 and 2) of 2-azabicyclo[2.2.2]octane were synthesized as semirigid analogues of dopamine (DA) to help evaluate the preferred conformation of dopamine at the uptake site of the presynaptic nerve terminal and at the DA receptor. Against the uptake of 0.1 microM [3H]DA by a synaptosomal preparation of corpus striatum from the reserpine-pretreated rat, 2 was found to have a weak inhibitory effect that was three times greater than that of 1 (IC50 = 32 vs. 110 microM). Interactions with DA receptors were assessed with competition for binding of [3H]apomorphine (APO) and on the effect on DA-sensitive adenylate cyclase. Compounds 1 and 2 were both virtually inactive against the binding of 0.5 nM [3H]APO at a screening concentration of 100 microM. The experimental compounds also exhibited only slight adenylate cyclase stimulation in rat striatal homogenates, with 1 appearing to be somewhat more active (at 50 or 400 microM). The weak activities of 1 and 2 and their relatively small differences in activity in these test systems suggest that the DA analogues interact only weakly with the DA transport and receptor sites, possibly as a result of the steric interference caused by the bulky bicyclic ring.

Adenylyl Cyclases↗

PubMed2005

Screening for colorectal cancer--now and the near future.

When used appropriately, screening for colorectal cancer (CRC) can reduce disease-related morbidity and mortality. Current methods include fecal occult blood testing (FOBT), flexible sigmoidoscopy [FS], barium enema, and colonoscopy; all are cost-effective techniques. Unfortunately, offering an array of options has not increased screening utilization, which continues to lag behind that of other common cancers. Newer techniques, particularly virtual colonoscopy (VC) and stool DNA testing, may offer attractive alternatives for healthcare provider recommendation and patient use.

Barium Sulfate↗

PubMedDate not supplied

A clinician's response to the oral contraceptive thrombosis controversy.

The controversy involving new progestin oral contraceptives (OC) began in late 1995, continued through 1996, and started to reach resolution in 1997. The fundamental question is whether OC containing desogestrel and gestodene have a different risk of thrombosis compared with OC containing older progestins. Correcting for preferential prescribing and the healthy user effect leads to the conclusion that all low-dose OC--regardless of progestin type--have an increased risk of venous thromboembolism. Low-dose OC do not increase the risk of myocardial infarction or stroke in healthy, non-smoking women less than 35 years of age. With effective patient screening for risk factors, the serious side effects of OC can be virtually eliminated.

Contraceptives, Oral, Combined↗

PubMed2005

Computer-aided detection for virtual colonoscopy.

Colon cancer is one of the leading causes of cancer deaths in the developed countries. Most colon cancers can be prevented if precursor colon polyps are detected and removed. Virtual colonoscopy, or CT colonography, has shown promise to be the future screening tool for polyp detection, with a number of studies performed at academic institutions showing high sensitivity and specificity. Two main factors limiting CT colonography in general use are its excessive interpretation time and the variable sensitivity among readers. This article discusses the potential of computer-aided detection to address these problems. We also review the current state of research in this field and the future roles and challenges of CAD for CT colonography.

Algorithms↗

PubMed2000

Diagnostic accuracy and interobserver agreement of CT colonography (virtual colonoscopy).

BACKGROUND AND AIMS: Computed tomographic (CT) colonography or virtual colonoscopy (VC) is a non-invasive imaging method proposed for screening patients with colorectal neoplasias. Our aims were to study the diagnostic accuracy and interobserver agreement of VC for correct patient identification compared with conventional colonoscopy (CC). METHODS: This was a prospective study of 50 patients successively undergoing VC and CC. Multiplanar two dimensional CT images and three dimensional VC were constructed using surface rendering software and interpreted by two independent investigator teams. VC findings were compared with those of CC. Interobserver agreement was determined using kappa statistics. RESULTS: CC found 65 polyps in 24 patients. For identification of patients with polyps > or =10 mm, the sensitivity of VC was 38% and 63%, and specificity was 74% and 74% for teams 1 and team 2. Interobserver agreement was good (kappa 0.72). For patients with polyps of any size, the sensitivity of VC was 75% and 71%, and specificity was 62% and 69% for teams 1 and 2. Interobserver agreement was fair (kappa 0.56). Accuracy improved when comparing the results of the first 24 with the last 26 patients. CONCLUSIONS: In our experience, VC had a low diagnostic value for identification of patients with colorectal neoplasias. Interobserver agreement for VC interpretation was fair. These results may be explained by software imperfections and a learning curve effect.

Aged↗

PubMedDate not supplied

Differential diagnosis of polypoid lesions seen at CT colonography (virtual colonoscopy).

Computed tomographic (CT) colonography, also referred to as virtual colonoscopy, holds significant promise for effective large-scale colorectal cancer screening. Two-dimensional (2D) and three-dimensional (3D) displays of the CT data are employed, both of which are critical for proper evaluation. Although many radiologists continue to use the 2D images for polyp detection, more emphasis on the 3D images for primary detection of polyps has yielded the best results for screening detection. The primary target lesion for colorectal screening is the adenomatous polyp, since detection and removal of all larger or advanced lesions could potentially prevent approximately 95% or more of all colon cancers. Frankly invasive adenocarcinoma is rarely encountered in an average-risk asymptomatic screening population, but it is of course another important target. In addition to these clinically significant epithelial neoplasms, however, a host of additional lesions and pseudolesions may be encountered that appear polypoid at CT colonography. A subset of "don't touch" lesions, which should not be confused with potential neoplasms, can also be recognized at CT colonography. A variety of useful techniques and observations can be used to increase the specificity of CT colonography for distinguishing false polyps from true polyps.

Colonic Polyps↗

PubMed2007

Monobodies: antibody mimics based on the scaffold of the fibronectin type III domain.

We developed the use of the 10th fibronectin type III domain of human fibronectin (FNfn10) as a scaffold to display multiple surface loops for target binding. We termed FNfn10 variants with novel binding function "monobodies." FNfn10 is a small (94 residues) protein with a beta-sandwich structure similar to the immunoglobulin fold. It is highly stable without disulfide bonds or metal ions, and it can be expressed in the correctly folded form at a high level in bacteria. These desirable physical properties render the FNfn10 scaffold compatible with virtually any display technologies. This chapter describes methods for library construction and screening and for the production of monobodies.

Amino Acid Sequence↗

PubMed1985

Plasmid fingerprinting. A tool for bacterial strain identification and surveillance of nosocomial and community-acquired infections.

Plasmid fingerprinting provides a rapid and dependable means of identifying bacterial isolates of the same strain. The stability, wide distribution, and diverse nature and size of extrachromosomal elements make it suitable for virtually all bacterial genera. There are many different procedures available for plasmid screening, and the one chosen depends primarily on the types of organisms to be analyzed. Some procedures are better suited to gram-positive organisms; others are better for visualizing the very large plasmids often seen in Pseudomonas and Rhizobium species. The key to the plasmid fingerprinting technique is agarose gel electrophoresis. In this step of the technique, it is important to differentiate open circular from closed circular forms of plasmids and to recognize the "smile effect." Plasmid fingerprinting can be utilized for epidemiologic studies of both nosocomial and community-acquired infections. The use of restriction endonuclease analysis can greatly enhance the ability of the investigator to differentiate strains that harbor only a single plasmid. Plasmid fingerprinting often provides the only differential characteristic for strains involved in epidemics.

Bacteria↗

PubMed1979

Rapid screening procedure for detection of plasmids in streptococci.

An enrichment procedure, yielding plasmid deoxyribonucleic acid preparations normally containing less than 5% chromosomal contamination, has been devised for the isolation of plasmids from virtually all species of streptococci. The procedure is rapid, reproducible, and inexpensive, requiring no radioisotopes or density gradient centrifugation. The procedure can be used for routine screening of several hundred isolates in a short period of time, and plasmids obtained from 10- to 20-ml cultures can readily be visualized in agarose gels.

DNA, Bacterial↗

PubMed2003

Virtual reality provides leisure time opportunities for young adults with physical and intellectual disabilities.

Due to limitations in their physical abilities, adults with cerebral palsy (CP) have relatively few opportunities to engage in independent leisure activities. This pervasive lack of opportunity often leads to the development of dependent behavioral patterns and learned helplessness. The objective of this pilot study was to explore ways in which virtual reality can provide positive and enjoyable leisure experiences during physical interactions with different game-like virtual environments and potentially lead to increased self-esteem and a sense of self-empowerment. The study sample consisted of five young male adults with CP and severe intellectual disabilities who are non-speaking and who use wheelchairs for mobility. Each participant experienced three game-like virtual scenarios via VividGroup's Gesture Xtreme video capture virtual reality (VR) system. The participant's video captured image was processed on the same plane as screen graphical animations that react in real time in response to his movements. Outcome measures included the participants' responses to a five-item presence questionnaire, a 6-item task specific questionnaire and observation of their videotaped performance while participating in the virtual games. Participants' responses to the questionnaire showed a high level of presence in all three scenarios. These participants demonstrated an exceptional degree of enthusiasm during each VR experience; some reacted to the various stimuli via appropriate and goal-oriented responses, in other cases the response was more arbitrary. Documentation of behaviors during and following the VR experiences may provide insight into the important role that VR may play in nurturing their self-esteem and sense of empowerment.

Adult↗

PubMed2003

Virtual colonoscopy using oral contrast compared with colonoscopy for the detection of patients with colorectal polyps.

BACKGROUND & AIMS: Virtual colonoscopy using abdominal spiral computed tomography scanning allows total colonic evaluation with minimal invasiveness. Two-dimensional images and selective 3-dimensional images of the colon are used to detect colorectal lesions. This trial used conventional colonoscopy with segmental unblinding to determine the ability of virtual colonoscopy to identify patients with colorectal lesions who need conventional colonoscopy. METHODS: We studied 205 patients with virtual colonoscopy using oral iodinated contrast preceding conventional colonoscopy. Colonic lavage was achieved with an oral sodium phosphosoda preparation and colonic distention with a carbon dioxide electronic insufflator. RESULTS: The overall sensitivity and specificity of virtual colonoscopy in identifying patients with colorectal lesions was 61.8% and 70.7%, respectively. Virtual colonoscopy was more accurate in identifying patients with lesions >/=6 mm (sensitivity 84.4% and specificity 83.1%) and those with lesions >/=10 mm (sensitivity 90% and specificity 94.6%). The negative predictive value of virtual colonoscopy was 95% for a 6-mm cutoff size and 98.9% for a 10-mm cutoff. Using a 10-mm cutoff, virtual colonoscopy precludes the need for conventional colonoscopy in 86% of patients with a 1% false-negative rate (68% with a 3.4% false-negative rate when using a 6-mm cutoff). CONCLUSIONS: Virtual colonoscopy has a high sensitivity and specificity for detecting patients with significant colorectal lesions. Its high negative predictive value may help reduce the number of negative screening colonoscopies. Further studies are needed to determine what lesion cutoff size is clinically acceptable and the appropriate interval time for repeat virtual colonoscopy when it detects lesions below this cutoff size.

Administration, Oral↗

PubMed2006

Colorectal cancer screening and surveillance.

Screening and surveillance substantially reduce both the incidence and mortality of colorectal cancer. Screening of normal-risk individuals may be accomplished by several methods, including fecal occult blood testing, fiberoptic sigmoidoscopy, double contrast barium enema, and colonoscopy. New technologies for screening are being developed, such as fecal immunochemical testing for blood,fecal DNA testing, and virtual colonoscopy. Patients at increased risk for colorectal cancer, such as those with a positive family history, previous adenomatous polyps or cancer, and inflammatory bowel disease, should be offered more intensive evaluation and surveillance.

Colorectal Neoplasms↗

PubMed1997

Association mapping of disease loci, by use of a pooled DNA genomic screen.

Genomic screening to map disease loci by association requires automation, pooling of DNA samples, and 3,000-6,000 highly polymorphic, evenly spaced microsatellite markers. Case-control samples can be used in an initial screen, followed by family-based data to confirm marker associations. Association mapping is relevant to genetic studies of complex diseases in which linkage analysis may be less effective and to cases in which multigenerational data are difficult to obtain, including rare or late-onset conditions and infectious diseases. The method can also be used effectively to follow up and confirm regions identified in linkage studies or to investigate candidate disease loci. Study designs can incorporate disease heterogeneity and interaction effects by appropriate subdivision of samples before screening. Here we report use of pooled DNA amplifications-the accurate determination of marker-disease associations for both case-control and nuclear family-based data-including application of correction methods for stutter artifact and preferential amplification. These issues, combined with a discussion of both statistical power and experimental design to define the necessary requirements for detecting of disease loci while virtually eliminating false positives, suggest the feasibility and efficiency of association mapping using pooled DNA screening.

Case-Control Studies↗

PubMed2004

Automatic identification of subcellular phenotypes on human cell arrays.

Light microscopic analysis of cell morphology provides a high-content readout of cell function and protein localization. Cell arrays and microwell transfection assays on cultured cells have made cell phenotype analysis accessible to high-throughput experiments. Both the localization of each protein in the proteome and the effect of RNAi knock-down of individual genes on cell morphology can be assayed by manual inspection of microscopic images. However, the use of morphological readouts for functional genomics requires fast and automatic identification of complex cellular phenotypes. Here, we present a fully automated platform for high-throughput cell phenotype screening combining human live cell arrays, screening microscopy, and machine-learning-based classification methods. Efficiency of this platform is demonstrated by classification of eleven subcellular patterns marked by GFP-tagged proteins. Our classification method can be adapted to virtually any microscopic assay based on cell morphology, opening a wide range of applications including large-scale RNAi screening in human cells.

Artifacts↗