Polymorphic microsatellites from sperm whales and their use in the genetic identification of individuals from naturally sloughed pieces of skin.
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Animals↗
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Identification of individual amino acids required for secretion within the haemolysin (HlyA) C-terminal targeting region.
The release of haemolysin from Escherichia coli involves direct secretion across both the inner and outer membranes. Secretion of HlyA is dependent upon a specific membrane export complex composed of HlyB, -D and possibly TolC. HlyA is targeted to the medium via the membrane translocation complex, by a novel C-terminal secretion signal. Previous studies involving deletion and fusion analyses have given contradictory results for the minimal length (20-60 residues) of this HlyA signal region and little is known of the nature of the specific residues and structural features required for function. In this study we have analysed, quantitatively, the effect upon secretion of many point mutations introduced into the HlyA C-terminus. The results indicate the presence of a minimal secretion signal domain whose proximal boundary extends to at least residue -46 and which contains at least four individual residues essential for maximal secretion levels. We propose that such residues act co-operatively, forming multiple contact points with the translocator proteins, with the 'best fit' promoting maximal levels of secretion.
Amino Acid Sequence↗
Biosynthesis of the ubiquinol-cytochrome c reductase complex in yeast. Characterization of precursor forms of the 44-kDa, 40-kDa and 17-kDa subunits and identification of individual messenger RNAs for these and other imported subunits of the complex.
The mitochondrial ubiquinol--cytochrome c reductase complex (complex III or cytochrome bc1 complex) is thought to consist of eight subunits, seven of which are specified by nuclear genes and synthesized in the cytoplasm. We have studied the synthesis of five of the nuclear-encoded subunits both in vivo and in vitro and show that of these the 44-kDa, 40-kDa and 17-kDa subunits are synthesized with cleavable extensions, while the 14-kDa and 11-kDa proteins are synthesized without detectable extra sequences. The sizes of the pre-sequences, as determined by the relative mobility of the precursor proteins in sodium dodecyl sulphate/polyacrylamide gels, range from 0.5-kDa for the 44-kDa and 40-kDa subunits to 9-kDa for the 17-kDa subunit. The existence in vivo of precursor forms to the 44-kDa, 40-kDa and 17-kDa subunits implies that import is at least partially a post-translational process. The precursor of the 44-kDa subunit can be processed post-translationally in vitro by isolated mitochondria. The messenger RNAs for subunits of the complex have been studied. Those coding for the 44-kDa, 40-kDa, 14-kDa and 11-kDa proteins and cytochrome c1 are of different sizes, indicating that each of these subunits is synthesized as a separate protein, rather than as part of a polyprotein precursor.
Chemical Phenomena↗
31P-NMR investigation of magnetically oriented rod outer segments. Spectral analysis and identification of individual phospholipids.
A 31P-NMR study of magnetically oriented bovine rod outer segments is presented. We demonstrate that carefully isolated bovine rod outer segments retain the capacity to orient in a magnetic field. Maximal orientation (85-90%) is achieved at field strengths over 4.7 T in the NMR spectrometer. The lineshape of the 'oriented spectra' is totally different from the 'bilayer lineshape' of randomly oriented photoreceptor membranes. The oriented spectra consist of two phospholipid peaks, a major low-field peak (75-80% of the total intensity) near 30 ppm, and a minor high-field peak near - 14 ppm as well as two sharp metabolite peaks around 0 ppm. The phospholipid peaks are a composite of three narrower partially resolved resonances assigned to the individual phospholipid classes phosphatidylserine, phosphatidylcholine and phosphatidylethanolamine. Based on the morphology and magnetic anisotropy of the rod outer segment, the major phospholipid peak is attributed to the flat part of the disk membranes while the phospholipids of the plasma membrane are thought to contribute only to the minor peak. Disk rim phospholipids and non-oriented material contribute to the minor peak and, in addition, contribute some intensity to the middle part of the spectrum. The phospholipid class composition of the major peak is estimated by spectral simulation and is consistent with the phospholipid class composition of rod outer segment membranes. Hence, 31P analysis of oriented rod outer segments resolves the main phospholipids in at least two different membrane pools in the rod outer segment and allows the differential investigation of these pools. Most of the mobile phosphate metabolite intensity resides in the Pi peak at 3.5 ppm. A slight shift in the Pi resonance position indicates a 0.2 pH unit acidification upon illumination of rhodopsin. The absence of detectable nucleotide resonances, when compared with chemical analysis, indicates that the majority of the nucleotide population present is rather immobile and probably bound to the membranes.
Animals↗
Usefulness of live/real time three-dimensional transthoracic echocardiography in the identification of individual segment/scallop prolapse of the mitral valve.
In this report, we present 34 patients in whom surgical intervention was undertaken for severe mitral insufficiency due to mitral valve prolapse (MVP). Location and severity of MVP and regurgitation were assessed preoperatively by live/real time three-dimensional transthoracic echocardiography and closely agreed with the surgical findings.
Adult↗
Proteins in fluid from non-keratinizing jaw cysts. 3. Identification of individual proteins with particular reference to alpha- and beta-globulins, including fibrinogen.
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Albumins↗
Identification of individual genotypes of Chlamydia trachomatis from experimentally mixed serovars and mixed infections among trachoma patients.
Genetic polymorphisms in the major outer membrane protein gene (omp-1) of Chlamydia trachomatis B and Ba serovars have been demonstrated in Tunisian isolates. A total of 15 of 27 unique sequence signatures or omp-1 genotypes were identified. However, differentiation of unique signatures from sequences that reflect those of strains involved in a mixed infection is necessary to define the molecular epidemiology of chlamydial ocular infections. We devised a strategy for identifying mixed infections by characterizing their effects on omp-1 genotyping. Various ratios of elementary bodies from organisms of serovars A, B, Ba, and C that cause trachoma were amplified by PCR and were subjected to automated and manual sequencing. Serovar-specific primers were also designed so that each serovar could be individually amplified and its omp-1 genotype unequivocally determined. One of 27 Tunisian samples showed a mixed infection with sequences comparable to those of serovars B and D. The omp-1 genotypes of organisms involved in mixed infections can be accurately identified by automated sequencing and will be useful for molecular epidemiologic studies of populations worldwide who live where trachoma is endemic.
Bacterial Outer Membrane Proteins↗
Identification of individual human immunodeficiency virus type 1 gp120 amino acids important for CD4 receptor binding.
The binding of the CD4 receptor by the human immunodeficiency virus type 1 gp120 exterior envelope glycoprotein is important for virus entry and cytopathic effect. To investigate the CD4-binding region of the gp120 glycoprotein, we altered gp120 amino acids, excluding cysteines, that are conserved among the primate immunodeficiency viruses utilizing the CD4 receptor. Changes in two hydrophobic regions (Thr-257 in conserved region 2 and Trp-427 in conserved region 4) and two hydrophilic regions (Asp-368 and Glu-370 in conserved region 3 and Asp-457 in conserved region 4) resulted in significant reductions in CD4 binding. For most of the mutations affecting these residues, the observed effects on CD4 binding did not apparently result from global conformational disruption of the gp120 molecule, as assessed by measurements of precursor processing, subunit association, and monoclonal antibody recognition. The two hydrophilic regions exhibit a strong propensity for beta-turn formation, are predicted to act as efficient B-cell epitopes, and are located adjacent to hypervariable, glycosylated regions. This study defines a small number of gp120 residues important for CD4 binding, some of which might constitute attractive targets for immunologic intervention.
Amino Acid Sequence↗
Regulation of the Discoidin I gamma gene in Dictyostelium discoideum: identification of individual promoter elements mediating induction of transcription and repression by cyclic AMP.
We dissected the promoter of the developmentally induced and cyclic AMP-repressed discoidin I gamma gene and identified a sequence element essential for developmental induction. Transfer of the element to an inactive heterologous promoter demonstrated that this sequence is sufficient to confer expression in axenically growing cells and to induce gene activity in development after growth on bacteria. A 16-base-pair sequence within this element was shown to be sufficient for induction in the discoidin promoter context and was used to reactivate different truncated promoter constructs. This led to the localization of an element necessary for down regulation of gene expression by extracellular cyclic AMP.
Base Sequence↗
Towards identification of individual homologous chromosomes: comparative genomic hybridization and spectral karyotyping discriminate between paternal and maternal euchromatin in Mus musculus x M. spretus interspecific hybrids.
We have developed an in situ technique to label individual euchromatic chromosome arms in interspecific crosses between Mus musculus (MMU) and M. spretus (MSP). The MMU and MSP genomes diverged 2-3 million years ago and show an overall sequence divergence of approximately 1%. Comparative hybridization of MMU versus MSP DNA and subsequent spectral analysis of the euchromatic hybridization profiles discriminated between maternal (MMU) and paternal (MSP) chromosomes in F(1) hybrids. Dispersed repetitive DNA elements were the preferred hybridization target of MMU DNA on maternal chromosomes and of MSP DNA on paternal chromosomes. Differences in centromeric satellite DNAs were detected by conventional fluorescence in situ hybridization and served as internal controls. Our experiments suggest that it is possible, in principle, to discriminate between paternal and maternal chromosomes on the basis of sequence differences.
Animals↗
Identification of individuals with covert ischemic thrombotic cerebrovascular disease: a discriminant function analysis.
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Age Factors↗
The relationship between endocytosis of concanavalin A and phytohaemagglutinin receptors and blast transformation, and direct identification of individual rabbit lymphocytes reactive to both mitogens.
Distribution and modulation of rabbit lymphocyte phytohaemagglutinin acceptors and concanavalin A acceptors during activation of rabbit lymphocytes have been examined by electron microscopy. Two types of cell surface acceptors have been tentatively identified, lectin binding acceptors that do not modulate, and receptors that are endocytosed when blast transformation is stimulated. All of the cells have binding acceptors for both lectins. Endocytosis correlates with early blast transformation and serves as an early marker for lymphocyte activation. When examined after 24 h of culture, those cells that undergo blast transformation contain endocytosed lectin receptors, whereas small untransformed cells do not. Capping prior to endocytosis is rarely observed. The mechanism whereby the signal for transformation is maintained after the reaction of lectin with cell surface receptors and transposed to the nucleus is not known. Although we conclude that endocytosis is an early event required for cell activation, it is possible that endocytosis is secondary to other activation events. By evaluation of sequential endocytosis, individual rabbit lymphocytes that endocytose only concanavalin A, only phytohaemagglutinin, both concanavalin A and phytohaemagglutinin, or neither lectin, have been identified.
Animals↗
Identification of individual barley chromosomes based on repetitive sequences: conservative distribution of Afa-family repetitive sequences on the chromosomes of barley and wheat.
The Afa-family repetitive sequences were isolated from barley (Hordeum vulgare, 2n = 14) and cloned as pHvA14. This sequence distinguished each barely chromosome by in situ hybridization. Double color fluorescence in situ hybridization using pHvA14 and 5S rDNA or HvRT-family sequence (subtelomeric sequence of barley) allocated individual barley chromosomes showing a specific pattern of pHvA14 to chromosome 1H to 7H. As the case of the D genome chromosomes of Aegilops squarrosa and common wheat (Triticum aestivum) hybridized by its Afa-family sequences, the signals of pHvA14 in barley chromosomes tended to appear in the distal regions that do not carry many chromosome band markers. In the telomeric regions these signals always placed in more proximal portions than those of HvRT-family. Based on the distribution patterns of Afa-family sequences in the chromosomes of barley and D genome chromosomes of wheat, we discuss a possible mechanism of amplification of the repetitive sequences during the evolution of Triticeae. In addition, we show here that HvRT-family also could be used to distinguish individual barley chromosomes from the patterns of in situ hybridization.
Biological Evolution↗
[A simplified screening test for the identification of individuals with diminished vision in developing countries].
Simple but effective tests have been produced for screening subjects with low vision in developing countries. These tests of distance and near vision, based on the E test, were evaluated and validated in trials with people aged 4-90 years, and have been field tested in the health, education and rehabilitation services in 32 developing countries. Their sensitivity and specificity as screening tools for low vision have been calculated: sensitivity of 85% and specificity of 96% for the distance vision test, and sensitivity of 100% and specificity of 84% for the near vision test. The content and format of the tests have been demonstrated to be appropriate for developing countries, and their effectiveness for screening for low vision has been confirmed.
Adolescent↗
Identification of individual acid mucopolysaccharide in tissue sections.
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Animals↗
Identification of individuals with insulin resistance using routine clinical measurements.
Insulin resistance is a treatable precursor of diabetes and potentially of cardiovascular disease as well. To identify insulin-resistant patients, we developed decision rules from measurements of obesity, fasting glucose, insulin, lipids, and blood pressure and family history in 2,321 (2,138 nondiabetic) individuals studied with the euglycemic insulin clamp technique at 17 European sites; San Antonio, Texas; and the Pima Indian reservation. The distribution of whole-body glucose disposal appeared to be bimodal, with an optimal insulin resistance cutoff of <28 micromol/min . kg lean body mass. Using recursive partitioning, we developed three types of classification tree models: the first, based on clinical measurements and all available laboratory determinations, had an area under the receiver operator characteristic curve (aROC) of 90.0% and generated a simple decision rule: diagnose insulin resistance if any of the following conditions are met: BMI >28.9 kg/m(2), homeostasis model assessment of insulin resistance (HOMA-IR) >4.65, or BMI >27.5 kg/m(2) and HOMA-IR >3.60. The fasting serum insulin concentrations corresponding to these HOMA-IR cut points were 20.7 and 16.3 microU/ml, respectively. This rule had a sensitivity and specificity of 84.9 and 78.7%, respectively. The second model, which included clinical measurements but no laboratory determinations, had an aROC of 85.0% and generated a decision rule that had a sensitivity and specificity of 78.7 and 79.6%, respectively. The third model, which included clinical measurements and lipid measurements but not insulin (and thus excluded HOMA-IR as well), had a similar aROC (85.1%), sensitivity (81.3%), and specificity (76.3%). Thus, insulin-resistant individuals can be identified using simple decision rules that can be tailored to specific needs.
Blood Glucose↗
Does the metabolic syndrome improve identification of individuals at risk of type 2 diabetes and/or cardiovascular disease?
OBJECTIVE: The metabolic syndrome has been promoted as a method for identifying high-risk individuals for type 2 diabetes and cardiovascular disease (CVD). We therefore sought to compare this syndrome, as defined by the National Cholesterol Education Program, to the Diabetes Predicting Model and the Framingham Risk Score as predictors of type 2 diabetes and CVD, respectively. RESEARCH DESIGN AND METHODS: A population-based sample of 1,709 initially nondiabetic San Antonio Heart Study (SAHS) participants were followed for 7.5 years, 195 of whom developed type 2 diabetes. Over the same time interval, 156 of 2,570 SAHS participants experienced a cardiovascular event. A population-based sample of 1,353 initially nondiabetic Mexico City Diabetes Study (MCDS) participants were followed for 6.5 years, 125 of whom developed type 2 diabetes. Baseline measurements included medical history, age, sex, ethnicity, smoking status, BMI, blood pressure, fasting and 2-h plasma glucose levels, and fasting serum total and HDL cholesterol and triglycerides. RESULTS: The sensitivities for predicting diabetes with the metabolic syndrome were 66.2 and 62.4% in the SAHS and the MCDS, respectively, and the false-positive rates were 27.8 and 38.7%, respectively. The sensitivity and false-positive rates for predicting CVD with the metabolic syndrome in the SAHS were 67.3 and 34.2%, respectively. At corresponding false-positive rates, the two predicting models had significantly higher sensitivities and, at corresponding sensitivities, significantly lower false-positive rates than the metabolic syndrome for both end points. Combining the metabolic syndrome with either predicting model did not improve the prediction of either end point. CONCLUSIONS: The metabolic syndrome is inferior to established predicting models for either type 2 diabetes or CVD.
Adult↗
Epidemiology of polyps in the rectum and sigmoid colon. Discriminant analysis for identification of individuals at risk of developing colorectal neoplasia.
In an endoscopic population screening examination for detection of colorectal polyps among 324 men and women aged 50-59 years, adenomas greater than or equal to 5 mm in diameter were found in 38 individuals, and 212 were polyp-free. Dietary registration was performed by 155 individuals on a case/control basis. At 2 years' follow-up study polyps less than 5 mm in diameter were also removed. This rendered a total of 39 adenoma patients and 59 polyp-free individuals available for a combined analysis of both case history and dietary information. Application of a linear discriminant model on the combined clinical and dietary information enabled us to select a possible set of variables to characterize 88 out of 98 individuals (90%) correctly as adenoma patients or polyp-free individuals. However, when only case history information was used for the total material of 38 adenoma patients and 212 polyp-free individuals, only 189 out of 250 individuals (76%) were correctly classified. Although age, smoking habits, and a family history of colorectal cancer appeared as useful variables in the discriminant function, this study demonstrated the importance of nutritional factors in characterization of adenoma patients in the present age group of 50-59 years. In the present study we could not find any set of case history information which could indicate the usefulness of a questionnaire-based 'pre-screening' to guide recommendations for endoscopic screening examination.
Colonic Polyps↗