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DNA aneuploidy and integration of human papillomavirus type 16 e6/e7 oncogenes in intraepithelial neoplasia and invasive squamous cell carcinoma of the cervix uteri.

PURPOSE: Increasingly deregulated expression of the E6-E7 oncogenes of high-risk human papillomaviruses (HR-HPVs) has been identified as the major transforming factor in the pathogenesis of cervical dysplasia and derived cancers. The expression of these genes in epithelial stem cells first results in chromosomal instability and induces chromosomal aneuploidy. It is speculated that this subsequently favors integration of HR-HPV genomes into cellular chromosomes. This in turn leads to expression of viral cellular fusion transcripts and further enhanced expression of the E6-E7 oncoproteins. Chromosomal instability and aneuploidization thus seems to precede and favor integration of HR-HPV genomes. EXPERIMENTAL DESIGN: To prove this sequential concept, we analyzed here the sequence of events of DNA aneuploidization and integration in a series of HPV-16-positive cervical dysplastic lesions and carcinomas. Eighty-five punch biopsies of HPV-16-positive cervical lesions (20 CIN1/2, 50 CIN3, and 15 CxCa) were analyzed for DNA ploidy by DNA flow cytometry and for integration of HPV E6/E7 oncogenes using the amplification of papillomavirus oncogene transcripts assay, a reverse transcription-PCR method to detect integrate-derived human papillomavirus oncogene transcripts. RESULTS: DNA aneuploidy and viral genome integration were both associated with increasing dysplasia (P < 0.001, chi(2) test for trend). In addition, DNA aneuploidy was associated with increased viral integration (P < 0.01, Fisher's exact test). Nineteen of 20 (95%) lesions with integrated viral genomes had aneuploid cell lines; however, only 19 of 32 (59%) lesions with aneuploid cell lines had integrated viral genomes. CONCLUSIONS: These data support the hypothesis that aneuploidization precedes integration of HR-HPV genomes in the progression of cervical dysplasia. Accordingly, deregulated viral oncogene expression appears to result first in chromosomal instability and aneuploidization and is subsequently followed by integration of HR-HPV genomes in the affected cell clones.

Adult↗

Analysis of integration activity of human immunodeficiency virus type-1 integrase.

The integration activity of human immunodeficiency virus type-1 (HIV-1) integrase was characterized in vitro by using pre-processed oligonucleotide substrates. The highest level of integration activity was found at pH 6.5 to 7.0, while the endonucleolytic activity was highest at pH 7.4 to 8.0. Although the endonucleolytic and integration reactions are consecutive in retroviral integration, our result indicates that the optimal conditions of the two reactions are quite different. In addition, it is suggested that the endonucleolytic and integration steps can be separated by control of the cellular physiological state in retroviral therapy. Strong integration was detected in the presence of 0.5-10 mM Mn2+ ion, but weak integration at around 10 mM Mg2+ ion. This observation explains that the Mn2+ ion is preferred to the Mg2+ ion as a cofactor in the integration reaction. Although there was no sequence-specificity in the integration site of the target DNA, integration was found to frequently occur at particular regions of the target DNA. Furthermore, the mutant integrases such as Asp116, Ser147, and Glu152, which had been reported previously, were shown to lose integration activity completely, indicating that these residues are critically involved in catalytic action.

Base Sequence↗

A comparison of the molecular structure of integrated hepatitis B virus genomes in hepatocellular carcinoma cells and hepatocytes derived from the same patient.

To elucidate critical genetic elements in the development of hepatocellular carcinoma associated with hepatitis B virus DNA integration, a single integrant in hepatocellular carcinoma cells and one species of multiple integrants in hepatocytes, both obtained from the same patient, were compared structurally using molecular cloning techniques. Both hepatitis B virus integrants showed similar inverted repeat sequences consisting of two defective virus genomes. The recombination of viral DNAs seemed to be mediated by short regions of base homology near the direct repeat 1 and at other regions of the virus genomes in both integrants. The virus component in the junction with host DNAs was the cohesive end region in each identical end of the viral integrant in hepatocellular carcinoma cells and in one end of the viral integrant in hepatocytes. The structure of the integrant in hepatocellular carcinoma cells was characterized by an inverted, duplicated conformation composed not only of integrated virus genomes but also of flanking cellular sequences. It was shown to be the so-called "alpha dimer" of satellite DNA. In contrast, the flanking, nonreiterated cellular DNA in the hepatocyte-derived clone did not show discernible rearrangement. These findings suggest that a common mechanism underlies the integration of hepatitis B virus DNA so that a similar organization of inverted repeat genomes is found in hepatocellular carcinoma cells and in hepatocytes. The unstable nature of cellular DNA where DNA integration occurs may be important in generating chromosome alterations found in hepatocellular carcinoma.

Adult↗

Modularity and parallel processing in the oculomotor integrator.

The neural signals that hold eye position originate in a brainstem structure called the neural integrator, so-called because it is thought to compute these position signals using a process equivalent to mathematical integration. Most previous experiments have assumed that the neural integrator reacts to damage like a single mathematical integrator: the eye is expected to drift towards a unique resting point at a simple exponential rate dependent on current eye position. Physiologically, this would require a neural network with uniformly distributed internal connections. However, Cannon et al. (1983) proposed a more robust modular internal configuration, with dense local connections and sparse remote connections, computationally equivalent to a parallel array of independent sub-integrators. Damage to some sub-integrators would not affect function in the others, so that part of the position signal would remain intact, and a more complex pattern of drift would result. We evaluated this parallel integrator hypothesis by recording three-dimensional eye positions in the light and dark from five alert monkeys with partial neural integrator failure. Our previous study showed that injection of the inhibitory gamma aminobutyric acid agonist muscimol into the mesencephalic interstitial nucleus of Cajal (INC) causes almost complete failure of the integrators for vertical and torsional eye position after approximately 30 min. This study examines the more modest initial effects. Several aspects of the initial vertical drift could not be accounted for by the single integrator scheme. First, the eye did not initially drift towards a single resting position; rapid but brief drift was observed towards multiple resting positions. With time after the muscimol injection, this range of stable eye positions progressively narrowed until it eventually approximated a single point. Second, the drift had multiple time constants. Third, multiple regression analysis revealed a significant correlation between drift rate and magnitude of the previous saccade, in addition to a correlation between drift rate and position. This saccade dependence enabled animals to stabilize gaze by making a series of saccades to the same target, each with less post-saccadic drift than its predecessor. These observations were predicted and explained by a model in which each of several parallel integrators generated a fraction of the eye-position command.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗

Dynamic and nonspecific dispersal of human T-cell leukemia/lymphoma virus type-I integration in cultured lymphoma cells.

The progression of HTLV-I proviral integration over a 3-year period of in vitro culture was examined in two human lymphoma lines, Hut 102 and MJ. Using specific HTLV-I molecular clones and a Southern analysis at different cell passages, Hut 102 increased from 2 to 19 integrated proviral integrations while MJ increased to at least 25 different integrations by passage 43. During the progress of increased superinfection and novel integration in vitro some of the previous proviral integrations were lost from the cultures. The 19 integrations of late passage Hut 102 cells were shown to be dispersed to 19 different human chromosomes by analysis of 34 distinct rodent X Hut 102 somatic cell hybrids which segregated human chromosomes (and included proviral integrations) in different combinations. The two primary integrations in Hut 102 were located on human chromosomes 4 and 20, respectively. A similar pattern of nonspecific integration was observed in somatic cell hybrid analysis of the 25 proviral integrations of MJ. The dynamic infection-reintegration process in vitro revealed in these studies may confuse experimental verification of potential cis acting functions of HTLV-I in the as yet poorly understood mechanism of neoplastic transformation.

Cell Line↗

Integration of Rous sarcoma virus DNA during transfection.

We have investigated the organization and integration sites of Rous sarcoma virus (RSV) DNA in NIH 3T3 mouse cells transformed by transfection with unintegrated and integrated donor RSV DNAs. RSV DNAs of different cell lines transformed by unintegrated donor DNA were flanked by different cellular DNA sequences, indicating that RSV DNA integrates at multiple sites during transfection. The RSV genomes of cells transformed by transfection were colinear with unintegrated RSV DNA, except that deletions within the terminal repeat units of RSV DNA were detected in some cell lines. These results suggested that the terminal repeat sequences of RSV DNA did not necessarily provide a specific integration site for viral DNA during transfection. In addition, cell lines transformed by integrated RSV DNAs contained both the RSV genomes and flanking cellular sequences of the parental cell lines, indicating that integration of integrated viral DNA during transfection occurred by recombinational events within flanking cellular DNA sequences rather than at the terminal of viral DNA. Integration of RSV DNA during transfection thus appears to differ from integration of RSV DNA in virus-infected cells, where the terminal repeat units of viral DNA provide a highly specific integration site. Integration of donor DNA during transfection of NIH 3T3 cells instead appears to proceed by a pathway which is nonspecific for both donor and recipient DNA sequences.

Animals↗

Analysis of transmural trend of myocardial integrated ultrasound backscatter for differentiation of hypertrophic cardiomyopathy and ventricular hypertrophy due to hypertension.

OBJECTIVES: This study was undertaken to differentiate hypertrophic cardiomyopathy from hypertensive hypertrophy using a newly developed M-mode format integrated backscatter imaging system capable of calibrating myocardial integrated backscatter with the power of Doppler signals from the blood. BACKGROUND: Myocardial integrated ultrasound backscatter changes in patients with hypertrophic cardiomyopathy; however, it is unknown whether ultrasound myocardial tissue characterization may be useful in differentiating hypertrophic cardiomyopathy from hypertensive hypertrophy. METHODS: Calibrated myocardial integrated backscatter and its transmural gradient were measured in the septum and posterior wall in 31 normal subjects, 13 patients with hypertensive hypertrophy and 22 patients with hypertrophic cardiomyopathy. The gradient in integrated backscatter was determined as the ratio of calibrated integrated backscatter in the endocardial half to that in the epicardial half of the myocardium. RESULTS: Cyclic variation of integrated backscatter was smaller and calibrated myocardial integrated backscatter higher in patients with hypertrophied hearts than in normal subjects, but there were no significant differences in either integrated backscatter measure between patients with hypertensive hypertrophy and those with hypertrophic cardiomyopathy. Transmural gradient in myocardial integrated backscatter was present only in patients with hypertrophic cardiomyopathy (5.0 +/- 1.8 dB [mean +/- SD] for the septum; 1.2 +/- 1.6 dB for the posterior wall). CONCLUSIONS: Hypertrophic cardiomyopathy and ventricular hypertrophy due to hypertension can be differentiated on the basis of quantitative analysis of the transmural gradient in integrated backscatter.

Acoustics↗

Electrocardiographic diagnosis of left ventricular hypertrophy by the time-voltage integral of the QRS complex.

OBJECTIVES: This study was conducted to test the hypothesis that the time-voltage integral of the QRS complex can improve the electrocardiographic (ECG) identification of left ventricular hypertrophy. BACKGROUND: Standard ECG criteria have exhibited poor sensitivity for left ventricular hypertrophy at acceptable levels of specificity. However, left ventricular mass may be more closely related to the time-voltage integral of the summed left ventricular dipole than to QRS duration or voltages used in standard ECG criteria. METHODS: Standard 12-lead ECGs, orthogonal lead signal-averaged ECGs and echocardiograms were obtained in 62 male control subjects without left ventricular hypertrophy and 51 men with left ventricular hypertrophy defined by echocardiographic criteria (indexed left ventricular mass > 125 g/m2). Voltage of the QRS complex was integrated over the total QRS duration in leads X, Y and Z to calculate the time-voltage integral of each orthogonal lead, of the maximal spatial vector complex and of the horizontal, frontal and sagittal plane vector complexes. RESULTS: At matched specificity of 99%, the 73% (37 of 51) sensitivity of the time-voltage integral of the vector QRS complex in the horizontal plane was significantly greater than the 10% sensitivity of the Romhilt-Estes point score, the 16% sensitivity of QRS duration alone, the 22% sensitivity of Cornell voltage, the 33% sensitivity of the 12-lead sum of QRS voltage and the 37% sensitivity of Sokolow-Lyon voltage (each p < 0.001). Sensitivity of the horizontal plane time-voltage integral was also greater than the 10% to 51% sensitivity of the time-voltage integral calculated in the individual X, Y or Z leads (p < 0.01 to < 0.001), the 18% and 35% sensitivity of the time-voltage integrals of the frontal and sagittal plane vectors (p < 0.001) and the 49% sensitivity of the time-voltage integral of the maximal spatial vector complex calculated from all three orthogonal leads (p < 0.001). Comparison of receiver operating characteristic curves confirmed that the superior performance of the horizontal plane time-voltage integral relative to standard and other signal-averaged criteria was independent of partition value selection. CONCLUSIONS: These findings suggest that use of the time-voltage integral of the QRS complex, a method that can be readily implemented on commercially available computerized ECG systems, can improve the accuracy of ECG methods for the identification of left ventricular hypertrophy.

Adult↗

Integrated backscatter for quantification of left atrial spontaneous echo contrast.

OBJECTIVES: This study was designed to develop a quantitative method of spontaneous echo contrast (SEC) assessment using integrated backscatter and to compare integrated backscatter SEC measurement with independent qualitative grades of SEC and clinical and echocardiographic predictors of thromboembolism. BACKGROUND: Left atrial SEC refers to dynamic swirling smokelike echoes that are associated with low flow states and embolic events and have been graded qualitatively as mild or severe. METHODS: We performed transesophageal echocardiography in 43 patients and acquired digital integrated backscatter image sequences of the interatrial septum to internally calibrate the left ventricular cavity and left atrial cavity under different gain settings. Patients were independently assessed as having no, mild or severe SEC. We compared intensity of integrated backscatter in the left atrial cavity relative to that in the left ventricular as well as to the independently assessed qualitative grades of SEC. Fourier analysis characterized the temporal variability of SEC. The integrated backscatter was compared with clinical and echocardiographic predictors of thromboembolism. RESULTS: The left atrial cavity integrated backscatter intensity of the mild SEC subgroup was 4.7 dB higher than that from the left ventricular cavity, and the left atrial intensity of the severe SEC subgroup was 12.5 dB higher than that from the left ventricular cavity. The left atrial cavity integrated backscatter intensity correlated well with the qualitative grade. Fourier transforms of SEC integrated backscatter sequences revealed a characteristic dominant low frequency/high amplitude spectrum, distinctive from no SEC. There was a close relationship between integrated backscatter values and atrial fibrillation, left atrial size, left atrial appendage flow velocities and thrombus. CONCLUSIONS: Integrated backscatter provides an objective quantitative measure of SEC that correlates well with qualitative grade and is closely associated with clinical and echocardiographic predictors of thromboembolism. The relationship between integrated backscatter measures and cardioembolic risk will be defined in future multicenter studies.

Aged↗

Functional integration in schizophrenia: too little or too much? Preliminary results on fMRI data.

The disconnectivity hypothesis proposes that schizophrenia results from poor or miswired anatomical connections. Theoretically, its functional counterpart should be disintegration. Integration is thought to allow segregated neurons to interact as a coherent whole, referred to as the "core", while the non-interacting part of the brain is referred to as the "rest". In this study, it is suggested that schizophrenia is the result of rest noise interfering with core activity. Two possible causes are assessed: (i) defective core integration, making the core more vulnerable to noise from the rest, or (ii) the rest being too highly integrated, meaning that it can interfere with the core. These hypotheses were tested using fMRI data acquired from 13 stabilized medicated schizophrenic subjects compared to 11 matched controls. Subjects were required to perform a series of lexical decision and retrieval tasks in separate sessions. The brain was divided into 90 components. Integration was defined as the amount of information shared between the components of a sub-system. An iterative aggregation procedure made it possible to identify a core on the basis of the functional clustering index, which assesses the integration of the core relative to its integration with the rest. Correlation of component-pairs within the core was also compared between the two groups. This procedure was repeated for each subject and for each task. Cores did not differ between the two groups, either in terms of integration or in terms of functional clustering index. However, the core was still highly integrated with the rest and the rest was overly integrated in schizophrenic subjects. Both anomalies were correlated with the negative symptoms. These findings were consistent regardless of the task considered. Furthermore, within the core, anterior-posterior correlations were lower in patients (between the frontal and the parietal and posterior cingulate cortices), whereas frontal left-right correlations were excessive. No significant correlation was found with the medication. Thus, it appears that schizophrenia entails a deleterious combination of too much "noisy" integration (from the rest) and too little "significant" integration (anterior-posterior functional connectivity).

Adult↗

Development-associated differences in integrative cartilage repair: roles of biosynthesis and matrix.

A recurring problem in tissue transplantation therapies for articular cartilage defects is the lack of integration between the implant and the host cartilage. Previous studies have shown that in vitro integration between explants of calf cartilage is markedly higher than that between fetal cartilage, despite similarly high levels of deposition of newly synthesized collagen. The aim of this study was to determine if cellular biosynthesis and extracellular matrix each contribute to these development-associated differences in integrative repair in vitro. The approach taken was to examine integration between specific combinations of cartilage explants that were apposed for two weeks. The cartilage matrix showed different propensities for repair, as integration of calf live cartilage to calf devitalized cartilage was greater than that of calf live cartilage to fetal devitalized cartilage. An inhibiting factor appeared to be present in fetal cartilage matrix since guanidine treatment of fetal devitalized cartilage was able to enhance its integration. The difference between integration to living cartilage and integration to devitalized cartilage, for calf and fetal tissue, indicated that the biosynthetic contribution to integration by calf cartilage was greater than the biosynthetic contribution by fetal cartilage. Thus, the increasing level of integration between fetal and fetal cartilage, fetal and calf cartilage, and calf and calf cartilage appeared to reflect both biosynthetic and matrix differences. Therapeutic strategies to enhance integration to cartilage may thus target both the extracellular components and the cellular biosynthetic activities of implants and host cartilage.

Animals↗

Analysis of junction sequences resulting from integration at nonhomologous loci in Neurospora crassa.

We have analyzed the junctions involved in two examples of ectopic integration of plasmids containing the am+ (glutamate dehydrogenase) gene into a strain of Neurospora crassa bearing a complete deletion of the am locus. In one transformed strain a single copy of plasmid DNA had been integrated into linkage group (LG) III DNA without the loss of chromosomal DNA. In contrast, 450 bp had been lost from plasmid sequences at the site of integration. The transforming DNA used was circular, so we postulate that the plasmid was linearized and truncated prior to its integration by end joining into a break in LG III DNA. There was no significant homology between the incoming DNA and DNA at the site of integration. The second transformed strain resulted from transformation with a linearized plasmid. It contained multiple integrated copies of plasmid DNA, one of which was recloned, together with adjacent chromosomal DNA, by plasmid rescue in Escherichia coli. Prior to integration into chromosomal DNA, the linear plasmid had been truncated by 64 bp on one end and 3.2 kbp on the other end. One end of the integrated DNA was adjacent to DNA from the right arm of LG I, while the other end was integrated into a copy of a repetitive sequence. Restriction fragment length polymerism mapping showed that integration was in a copy of the repetitive sequence that is linked to the previously unassigned telomere M11 and is distantly linked to the LG VI marker con-11. Genetic analysis revealed that a long segment of LG I containing all markers from un-1 to the right tip has been translocated to the right end of LG VI. Tetrad analysis showed that the integrated DNA was closely linked to the translocation. We conclude that the transforming DNA was truncated and joined to DNA from two different chromosomes by end joining during the formation of a quasiterminal translocation, T(IR----VIR) UK-T12. We also conclude that the previously unassigned telomere, M11, is the right end of LG VI.

Base Sequence↗

Research environments that promote integrity.

BACKGROUND: The body of empirical knowledge about research integrity and the factors that promote research integrity in nursing research environments remains small. OBJECTIVE: To propose an internal control model as an innovative framework for the design and structure of nursing research environments that promote integrity. METHODS: An internal control model is adapted to illustrate its use for conceptualizing and designing research environments that promote integrity. RESULTS: The internal control model integrates both the organizational elements necessary to promote research integrity and the processes needed to assess research environments. The model provides five interrelated process components within which any number of research integrity variables and processes may be used and studied: internal control environment, risk assessment, internal control activities, monitoring, and information and communication. DISCUSSION: The components of the proposed research integrity internal control model proposed comprise an integrated conceptualization of the processes that provide reasonable assurance that research integrity will be promoted within the nursing research environment. Schools of nursing can use the model to design, implement, and evaluate systems that promote research integrity. The model process components need further exploration to substantiate the use of the model in nursing research environments.

Communication↗

Identification of Arabidopsis thaliana transformants without selection reveals a high occurrence of silenced T-DNA integrations.

Several recent investigations of T-DNA integration sites in Arabidopsis thaliana have reported 'cold spots' of integration, especially near centromeric regions. These observations have contributed to the ongoing debate over whether T-DNA integration is random or occurs preferentially in transcriptionally active regions. When transgenic plants are identified by selecting or screening for transgenic activity, transformants with integrations into genomic regions that suppress transcription, such as heterochromatin, may not be identified. This phenomenon, which we call selection bias, may explain the perceived non-random distribution of T-DNA integration in previous studies. In order to investigate this possibility, we have characterized the sites of T-DNA integration in the genomes of transgenic plants identified by pooled polymerase chain reaction (PCR), a procedure that does not require expression of the transgene, and is therefore free of selection bias. Over 100 transgenic Arabidopsis plants were identified by PCR and compared with kanamycin-selected transformants from the same T(1) seed pool. A higher perceived transformation efficiency and a higher frequency of transgene silencing were observed in the PCR-identified lines. Together, the data suggest approximately 30% of transformation events may result in non-expressing transgenes that would preclude identification by selection. Genomic integration sites in PCR-identified lines were compared with those in existing T-DNA integration databases. In PCR-identified lines with silenced transgenes, the integration sites mapped to regions significantly underrepresented by T-DNA integrations in studies where transformants were identified by selection. The data presented here suggest that selection bias can account for at least some of the observed non-random integration of T-DNA into the Arabidopsis genome.

Arabidopsis↗

Auditory temporal integration and the power function model.

The auditory temporal integration function was studied with the objective of improving both its quantitative description and the specification of its principle independent variable, stimulus duration. In Sec. I, temporal integration data from 20 studies were subjected to uniform analyses using standardized definitions of duration and two models of temporal integration. Analyses revealed that these data were best described by a power function model used in conjunction with a definition of duration, termed assigned duration, that de-emphasized the rise/fall portions of the stimuli. There was a strong effect of stimulus frequency and, in general, the slope of the temporal integration function was less than 10 dB per decade of duration; i.e., a power function exponent less than 1.0. In Sec. II, an experimental study was performed to further evaluate the models and definitions. Detection thresholds were measured in 11 normal-hearing human subjects using a total of 24 single-burst and multiple-burst acoustic stimuli of 3.125 kHz. The issues addressed are: the quantitative description of the temporal integration function; the definition of stimulus duration; the similarity of the integration processes for single-burst and multiple-burst stimuli; and the contribution of rise/fall time to the integration process. A power function in conjunction with the assigned duration definition was again most effective in describing the data. Single- and multiple-burst stimuli both seemed to be integrated by the same central mechanism, with data for each type of stimulus being described by a power function exponent of approximately 0.6 at 3.125 kHz. It was concluded that the contribution of the rise/fall portions of the stimuli can be factored out from the rest of the temporal integration process. In Sec. III, the conclusions that emerged from the review of published work and the present experimental work suggested that auditory temporal integration is best described by a power function in conjunction with the assigned duration definition. The exponent for the power function is typically less than 1.0, and varies with frequency and hearing level. Second, a means of empirically assaying the contribution of the rise-fall portions of the stimuli is presented and evaluated. Finally, properties of a central auditory integrator are hypothesized.(ABSTRACT TRUNCATED AT 250 WORDS)

Adult↗

Measures of auditory-visual integration in nonsense syllables and sentences.

For all but the most profoundly hearing-impaired (HI) individuals, auditory-visual (AV) speech has been shown consistently to afford more accurate recognition than auditory (A) or visual (V) speech. However, the amount of AV benefit achieved (i.e., the superiority of AV performance in relation to unimodal performance) can differ widely across HI individuals. To begin to explain these individual differences, several factors need to be considered. The most obvious of these are deficient A and V speech recognition skills. However, large differences in individuals' AV recognition scores persist even when unimodal skill levels are taken into account. These remaining differences might be attributable to differing efficiency in the operation of a perceptual process that integrates A and V speech information. There is at present no accepted measure of the putative integration process. In this study, several possible integration measures are compared using both congruent and discrepant AV nonsense syllable and sentence recognition tasks. Correlations were tested among the integration measures, and between each integration measure and independent measures of AV benefit for nonsense syllables and sentences in noise. Integration measures derived from tests using nonsense syllables were significantly correlated with each other; on these measures, HI subjects show generally high levels of integration ability. Integration measures derived from sentence recognition tests were also significantly correlated with each other, but were not significantly correlated with the measures derived from nonsense syllable tests. Similarly, the measures of AV benefit based on nonsense syllable recognition tests were found not to be significantly correlated with the benefit measures based on tests involving sentence materials. Finally, there were significant correlations between AV integration and benefit measures derived from the same class of speech materials, but nonsignificant correlations between integration and benefit measures derived from different classes of materials. These results suggest that the perceptual processes underlying AV benefit and the integration of A and V speech information might not operate in the same way on nonsense syllable and sentence input.

Adult↗

Development of an in vitro integration assay for the Bacteroides conjugative transposon CTnDOT.

Integrated self-transmissible elements called conjugative transposons (CTns) are responsible for the transfer of antibiotic resistance genes in many different species of bacteria. One of the best characterized of these newly recognized elements is the Bacteroides CTn, CTnDOT. CTnDOT is thought to have a circular transfer intermediate that transfers to and integrates into the genome of the recipient cell. Previous investigations of the mechanism of CTnDOT integration have been hindered by the lack of an in vitro system for checking this model of integration and determining whether the CTnDOT integrase alone was sufficient to catalyze the integration reaction or whether host factors might be involved. We report here the development of an in vitro system in which a plasmid containing the joined ends of CTnDOT integrates into a plasmid carrying a CTnDOT target site. To develop this in vitro system, a His-tagged version of the integrase gene of CTnDOT was cloned and shown to be active in vivo. The protein produced by this construct was partially purified and then added to a reaction mixture that contained the joined ends of the circular form of CTnDOT and a plasmid carrying one of the CTnDOT target sites. Integration was demonstrated by using a fairly simple mixture of components, but integration was stimulated by a Bacteroides extract or by purified Escherichia coli integration host factor. The results of this study demonstrate both that the circular form of CTnDOT is the form that integrates into the target site and that host factors are involved in the integration process.

Bacteroides↗

Structural integrity of resin-modified glass ionomers as affected by the delay or omission of light activation.

Since light activation of resin-modified glass ionomers as a means of polymerizing the HEMA is usually done shortly after mixing occurs, the acid-base reaction will proceed mainly within a formed HEMA-polymer matrix. Delaying or omitting light activation may alter the structure and consequently its integrity. The aim of this study was to investigate the effect on the structural integrity of Fuji II LC, Photac-Fil, and Vitremer by delaying or omitting light initiation as compared with the integrity when light activation is performed 2 min after mixing occurs. We evaluated integrity by three-body wear experiments, conducted 8 hrs after sample preparation, to establish the integrity in the early phase of hardening, as well as after 1 wk and after 4 mos, to follow the materials throughout the process of maturation. When light activation was delayed for 1 hr, the structural integrity of Fuji II LC and Photac-Fil improved significantly in the early stages of hardening. In the case of Vitremer, an hour's delay of light activation significantly decreased integrity, which declined further when light activation was omitted. Fuji II LC was not affected by the omission of light activation, while Photac-Fil was markedly weakened. After 4 mos of aging, most of the samples of each product which had been cured by the different methods attained equal integrity, with the exception of the non-light-activated Vitremer samples, which remained weaker. We concluded that the structural integrity of resin-modified glass-ionomer cements benefits from a chemical integration of the polyalkenoate and poly-HEMA networks, as in Vitremer. Improvement in the structural integrity in the early phase for cements with a mechanical entanglement of the matrices, as in Fuji II LC and Photac-Fil, requires an acid-base reaction, a considerable portion of which may take place before activation of the HEMA polymerization.

Composite Resins↗