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Plasma membrane integrity of cryopreserved human sperm: an investigation of the results of the hypoosmotic swelling test, the water test, and eosin-Y staining.

OBJECTIVE: [1] To examine the relationship between sperm membrane integrity and motion parameters before and after cryopreservation; [2] to determine the capacity of the membrane integrity tests to predict the outcome of cryopreservation in fertile and infertile men; and [3] to examine the degree of agreement between tail and head membrane integrity of testicular and ejaculated immotile sperm cryopreserved for intracytoplasmic sperm injection. DESIGN: Prospective study. SETTING: Academic tertiary care institution. PATIENT(S): Fertile donors and normozoospermic oligozoospermic, and asthenozoospermic subfertile men. INTERVENTION(S): Semen samples were cryopreserved and thawed for analysis. MAIN OUTCOME MEASURE(S): Sperm membrane integrity and computer-assisted motion parameters. RESULT(S): The hypoosmotic swelling test and water test had a significant and positive correlation in the fresh and cryopreserved ejaculates of all groups. The results of the hypoosmotic swelling test correlated positively with the percent motility in the fresh ejaculates of fertile and subfertile men. None of the membrane integrity tests correlated with the cryosurvival rate in any group. In the ejaculated and testicular samples with no postcryopreservation motility, the simultaneous assessment of hypoosmotic swelling test and eosin showed that of 33% sperm exhibiting coiling with the hypoosmotic swelling test, only 9% were eosin negative, whereas 24% were eosin positive. CONCLUSION(S): [1] The water test may be a simpler replacement for the hypoosmotic swelling test; [2] none of the membrane integrity tests predicted sperm motility after cryopreservation; and [3] there was a high degree of disagreement between the hypoosmotic swelling test and eosin in the samples with no postcryopreservation motility.

Cell Membrane↗

Effects of a benzodiazepine, lorazepam, on motion integration and segmentation: an effect on the processing of line-ends?

Previous studies have shown that the perceptual integration of component motions distributed across space is inhibited whenever segmentation cues, such as line-ends, are salient. Herein, we investigate to what extent enhanced inhibition induced by lorazepam, a benzodiazepine facilitating the fixation of GABA on GABAA receptors, modifies the balance between motion integration and motion segmentation at the behavioural level. Motion integration was tested in 16 healthy volunteers taking a single and oral dose of either placebo or lorazepam (0.038 mg kg-1). The stimulus consisted of an outlined diamond presented behind four, otherwise invisible, apertures and translating along a circular trajectory (Lorenceau & Shiffrar (1992). Vision Research, 32, 263-273). Under these conditions, recovering the global diamond direction requires the integration of the component motions available within each aperture. The observers were asked to discriminate the global, clockwise or counter-clockwise, diamond direction under difficult--at high luminance contrasts--or easy--at low luminance contrasts--conditions. Overall, reaction times and error rates increased in the lorazepam group as compared to the placebo group, suggesting strong non-specific effects. However, the changes in performance in the lorazepam group are not homogeneous across conditions, suggesting that lorazepam also induces specific effects that modulate the integration/segmentation balance. Additional experiments performed with visible apertures or visible diamond vertices indicate that the effects of lorazepam are unlikely to reflect a deficit of motion processing or motion integration mechanisms since performance is only slightly impaired in the lorazepam as compared to the placebo group under these conditions. These results suggest that lorazepam might specifically modulate the saliency of line-ends, presumably because processing these features involves inhibitory mechanisms using GABA as a neuromediator, and in turn modify the balance between motion integration and segmentation.

Anti-Anxiety Agents↗

Contour integration in color vision: a common process for the blue-yellow, red-green and luminance mechanisms?

We compare the performance of the red-green, blue-yellow and luminance postreceptoral mechanisms on a contour integration task requiring the linking of oriented Gabor elements across space to extract a winding 'path' or contour. We first establish that for all three mechanisms curvature and contrast are independent; losses in performance due to one cannot be compensated by changes in the other. We then compare contour integration by the three mechanisms using a method that controls for their differences in cone contrast thresholds. Our results show that despite the poor orientation discrimination thresholds and poor spatial sampling found for the blue-yellow mechanism, all three mechanisms perform similarly on contour integration over a wide range of curvatures. Furthermore, all three mechanisms have the same dependence on path curvature. We also investigate the effects of adding external orientation noise. Our results imply that the internal orientation noise for extracting 'aligned' path elements is similar in the three mechanisms and for all path curvatures, and the relative efficiencies are also similar for the three mechanisms. To account for our results, we propose that the three postreceptoral mechanisms use a common contour integration process. This linking process, however, cannot be color-blind; our last experiment shows that linking between different chromatic mechanisms or between opposite spatial phases disrupts contour integration. We thus propose that the common integration process remains sensitive to the color contrast and phase of its inputs.

Color Perception↗

Mechanosensitivity as an integrative system in heart: an audit.

This review examines a manifold of apparently loosely linked observations and mechanisms, from membrane to man, and assembles them to support the notion that mechanoelectric transduction is an integrative regulatory system in the heart. For this, the assemblage has to satisfy, at least to some extent, criteria that apply to other integrative regulatory systems such as the endocrine and nervous systems. The integrative effectors in the endocrine system are chemical linkages, circulating hormones: in the nervous system the linkage is a network of cables, nerve conduction and neurotransmitters. Mechanical integration is would be effected through mechanical machinery, cardiac contractile and hydraulic function with attendant stress and strain transmitted via "tensegrity". This can, through the cytoskeleton, begin with membrane integrins and transmit intracellularly for example via F actins to reach the rest of the membranous integrins. Further transmission to the organ is via cell-to-cell adhesion complexes and the extracellular matrix. This tensegrity facilitates integration of force and strain changes from area to area. In consequence, and analogous to the neurendocrine system, mechanoelectric transduction should, and does (1) operate at the molecular or membrane level--this would be via mechanotransducers affecting transmembrane ionic flow; (2) operate in the cell--to influence electrophysiology; (3) have a multicellular expression--e.g. mechanical distortion of one cell can raise intracellular calcium of an adjacent cell; (4) express in the intact organ--e.g. an increase in venous return hydraulically distends the sinoatrial node, steepening its pacemaker potential, thus increasing heart rate. It should also (5) demonstrate elements of a feedback system--"mechanoelectric feedback", and (6) interact with other systems--the cytoskeleton incorporates cell signalling complexes intersecting with other signal cascades. Finally, (7) it can malfunction to produce clinical abnormality--it contributes electrophysiologically to lethal cardiac arrhythmia. This anatomical and functional behaviour of mechanoelectric transduction could sanction the prospect of viewing it as analogous to the other integrative physiological systems.

Actins↗

Schizophrenia and the mechanisms of conscious integration.

This article considers the possibility that defective interactions among distributed brain areas may underlie certain dysfunctions of conscious integration such as those seen in schizophrenia. Recent experimental evidence obtained using whole-head magnetoencephalography during binocular rivalry is first reviewed. The results outline a cortical network that underlies conscious integration in the normal brain. This network is not localized to a small part of the brain but it is distributed over frontal, parietal, temporal, and occipital areas. Large-scale simulations of the dynamics of thalamocortical integration are then examined. These studies indicate that several factors can affect the rapid integration of the activity of distributed thalamocortical regions and the resulting behavioral performance. These simulations show that an altered dynamics of corticothalamic and corticocortical re-entrant circuits can result from increased conduction delays, blockade of voltage-dependent connections, reduced synaptic density, and disruptions of the local connectivity within a single cortical area. It can also result from alterations in the activity of diffuse ascending systems that lead to defective reinforcement of integrated activity patterns. Finally, the article briefly reviews theoretical measures of the integration of multiple brain areas, such as measures of functional clustering. These measures have been applied to PET data obtained from schizophrenic subjects and controls while performing cognitive tasks. The results show a change in the functional interactions among distributed brain areas in schizophrenics despite the absence of a change in activation patterns. The possibility is raised that disruption of re-entrant interactions among cortical areas may contribute to the pathophysiology of schizophrenia.

Brain↗

DNA integrity determination in marine invertebrates by Fast Micromethod.

This study was focused toward the adaptation of the previously developed Fast Micromethod for DNA damage determination to marine invertebrates for the establishment of biomonitoring assessment. The Fast Micromethod detects DNA damage (strand breaks, alkali-labile sites and incomplete excision repair) and determines DNA integrity in cell suspensions or tissue homogenates in single microplates. The procedure is based on the ability of the specific fluorochrome dye PicoGreen to preferentially interact with high integrity DNA molecules, dsDNA, in the presence of ssDNA and proteins in high alkaline medium, thereby allowing direct fluorometric measurements of dsDNA denaturation without sample handling and stepwise DNA separations. The results presented herein describe the influence of the DNA amount and the pH of the denaturation media on slopes of the kinetic denaturation curves and calculated strand scission factors (SSFs). The optimal amount of DNA in Mytilus galloprovincialis gills homogenate was found to be 100 ng ml(-1) and the greatest differences in DNA unwinding kinetics (slopes and SSF values) were reached at pH 11.5. The induction of DNA damage and loss of DNA integrity was measured in native DNA isolated from cotton-spinner Holothuria tubulosa, marine sponge Suberites domuncula cells and mussel M. galloprovincialis gills homogenate. DNA damage and loss of DNA integrity were detected after induction by different doses of (gamma-rays, generated by 137Cs 1800 Ci; 0-500 rad in marine sponge S. domuncula cells up to SSFx(-1) values 0.082 +/- 0.012 for the highest radiation dose). Analysis by chemical xenobiotics based on the in vitro action of bleomycin (bleomycin-Fe(II) complex 0-50 or 0-83 microg ml(-1) (microM)) with native DNA from cotton-spinner H. tubulosa and mussel M. galloprovincialis gills homogenate yielded values of 0.537 +/- 0.072 and 0.130 +/- 0.018, respectively. In vivo experiments with mussel M. galloprovincialis gills homogenate by 4-nitroquinoline-N-oxide (NQO; 0-1 microg g(-1) NQO mussel) and benzo[a]pyrene (B[a]P; 0-20 microg g(-1) B[a]P mussel) indicated SSFx(-1) values of 0.121 +/- 0.016 and 0.090 +/- 0.007, respectively, for the highest applied doses of chemical xenobiotics. The analytical technique described here allows simple and fast analysis of DNA integrity, requires very short time for multiple analyses (less than 3 h) and even less than 100 ng DNA per single well (50 ng DNA isolated from cotton-spinner, 12,500 sponge cells or about 10 mg of mussel gills homogenate) in a microplate. This makes the Fast Micromethod applicable for the measurement of DNA integrity of small samples for genotoxicity assessment (biomonitoring), the effects of genotoxins on lower marine taxa or sessile invertebrates in marine environment (e.g. sponges, mussels) and the estimation of directional changes and harmful effects in the ecosystem.

Animals↗

Community and self: concepts for rural physician integration and retention.

Low physician availability in rural areas of the United States is a long-standing issue. While most research has focused upon the locational decisions of physicians to understand the causes of the problem, a relatively new and neglected research focus is the retention of rural physicians. This paper takes the perspective that integration of physicians within rural communities is the basis for retention. It introduces concepts of self and community as a basis for understanding "domains" of rural physician integration. From data collected during in-depth qualitative research in rural Kentucky, a framework of integration domains is developed. The three domains of physician self, medical community, and community-at-large are elaborated with examples from the data. To further explain the relationships among the integration domains and their value, the findings are combined with three concepts from social theory. Social capital, core participation, and community reconstruction add dynamic and meaningful aspects to the context in which integration and retention occur, and they provide a conceptual basis to use when investigating or devising actions to facilitate integration.

Attitude to Health↗

Temporal integration in a subjective accentuation task as a function of child cognitive development.

We have previously shown that temporal integration in the domain of a few seconds may be studied using a subjective accentuation paradigm. Here we report developmental effects on the limits of this temporal integration in 9-10-year-olds in comparison with 13-14-year-olds. The task was to listen to a string of identical metronome beats and mentally bind the presented beats by subjectively accentuating every second, third or nth beat. The integration interval length was defined as the number of stimuli mentally connected multiplied by the temporal interval between two successive beats. For the lowest stimulus frequency integration intervals were approximately 3 s for the older and 2.2 s for the younger children. For higher frequencies integration intervals got systematically shorter, but being always longer for the older age group. It is suggested that the prefrontal region is responsible for this developmental effect. The expansion of temporal integration correlates with cognitive development in the investigated phase of ontogenesis.

Adolescent↗

pBRINT-Ts: a plasmid family with a temperature-sensitive replicon, designed for chromosomal integration into the lacZ gene of Escherichia coli.

A pBRINT-Ts family of integrative vectors for Escherichia coli was constructed by using a temperature-sensitive replicon derived from pSC101, a region of homology to the lacZ gene, and various antibiotic resistance markers (kanamycin, chloramphenicol and gentamycin) for selection of the integrants. The gene or group of genes to be integrated can be inserted into a multiple cloning site, flanked by an antibiotic resistance marker and lacZ sequences. A simple and rapid procedure was developed for the selection of cells where allelic exchange had occurred. With this procedure, the efficiency of integration of around 10-3 was observed, using several E. coli strains. From colonies with an integrated pBRINT-Ts plasmid, we detected an average allelic exchange event frequency of 7.5%. As a test for this system, we integrated the amy gene that codes for the alpha-amylase from B. stearothermophilus, into the lacZ gene of E. coli W3110. Production of alpha-amylase was found to be proportional to copy number; at up to 10 copies per chromosome.

Chromosomes, Bacterial↗

Developing an integrative therapies program in a tertiary care cardiovascular hospital.

This article describes one hospital's approach to developing an inpatient integrative therapies program and the foundation for a broader integrative healthcare vision. Since the program's inception, additional evidence has accumulated in the literature supporting the impact of integrative therapies strategies on patients' quality of life during inpatient stays. These findings and our own evaluation processes have encouraged continued program growth. Several elements were critical to the program's success. Review of the literature and investigation of other programs served as preparation before the actual program started. It was necessary to have administrative, nursing, and physician champions who were able to envision the program and see the value of this approach for patients. We appreciated the need for evidence-based outcomes research and demonstrable patient outcomes. Finally, a program manager was hired who was able to understand the culture of the hospital and the organizational change process. Each of these basic steps, which called for interdisciplinary collaboration, allowed us to accomplish the goal of using integrative therapies as adjuncts to conventional medical care and thereby supported an integrative approach. Consistently linking the integrative vision to patient needs and requirements helped us to identify many new avenues to expand upon this work. The process of program development described may be useful to other inpatient cardiovascular programs inclusive of critical care settings. Adaptations of our experience to other populations in critical care and across other hospital settings may be possible.

Cardiac Care Facilities↗

Molecular dynamics integration meets standard theory of molecular vibrations.

An iterative SISM (split integration symplectic method) for molecular dynamics (MD) integration is described. This work explores an alternative for the internal coordinate system prediction in the SISM introduced by JaneZic et al. (J. Chem. Phys. 2005, 122, 174101). The SISM, which employs a standard theory of molecular vibrations, analytically resolves the internal high-frequency molecular vibrations. This is accomplished by introducing a translating and rotating internal coordinate system of a molecule and calculating normal modes of an isolated molecule only. The Eckart frame, which is usually used in the standard theory of molecular vibrations as an internal coordinate system of a molecule, is adopted to be used within the framework of the second order generalized leapfrog scheme. In the presented MD integrator the internal coordinate frame at the end of the integration step is predicted halfway through the integration step using a predictor-corrector type iterative approach thus ensuring the method's time reversibility. The iterative SISM, which is applicable to any system of molecules with one equilibrium configuration, was applied here to perform all-atom MD simulations of liquid CO2 and SO2. The simulation results indicate that for the same level of accuracy, this algorithm allows significantly longer integration time steps than the standard second-order leapfrog Verlet (LFV) method.

Journal Article↗

Cre/lox-mediated site-specific integration of Agrobacterium T-DNA in Arabidopsis thaliana by transient expression of cre.

The Cre/lox system was used to obtain targeted integration of an Agrobacterium T-DNA at a lox site in the genome of Arabidopsis thaliana. Site-specific recombinants, and not random events, were preferentially selected by activation of a silent lox-neomycin phosphotransferase (nptII) target gene. To analyse the effectiveness of Agrobacterium-mediated transfer we used T-DNA vectors harbouring a single lox sequence (this vector had to circularize at the T-DNA left- and right-border sequences prior to site-specific integration) or two lox sequences (this vector allowed circularization at the lox sequences within the T-DNA either prior to or after random integration, followed by targeting of the circularized vector), respectively. Furthermore, to control the reversibility of the integration reaction, Cre recombinase was provided transiently by using a cotransformation approach. One precise stable integrant was found amongst the recombinant calli obtained after transformation with a double-lox T-DNA vector. The results indicate that Agrobacterium-mediated transformation can be used as a tool to obtain site-specific integration.

Arabidopsis↗

The integration of treatment and rehabilitation in psychiatric practice and services: a case study of a community mental health center.

After briefly reviewing the relationship of psychosocial rehabilitation to psychiatric practice, the authors recommend a renewed commitment of psychiatrists to bridge and integrate psychiatric treatment with psychosocial rehabilitation in practice and in the organization of services. They use the case example of an urban, community mental health center to illustrate a strategy for achieving greater integration of these two, relatively independent fields of professional practice. The Center's strategy for integration includes (1) center-wide planning, (2) structuring the medical staff office to support the task of integration, (3) establishing a model of practice and principles of care that supports both domains of intervention, (4) educating medical staff about psychosocial rehabilitation, (5) inter-disciplinary team building, including a definition and discussion of professional roles, (6) expanding services research on psychosocial rehabilitation, and (7) advocating in alliance with rehabilitation colleagues for expanded psychosocial rehabilitation services and their integration with treatment. By taking initiative to forward the integration of treatment and rehabilitation, psychiatrists better serve seriously ill patients and more effectively define their own work and roles.

Combined Modality Therapy↗

Target sites for SINE integration in Brassica genomes display nuclear matrix binding activity.

Short interspersed nuclear elements (SINEs) are ubiquitous components of complex animal and plant genomes. SINEs are believed to be important players in eukaryotic genome evolution. Studies on SINE integration sites have revealed non-random integration without strict nucleotide sequence requirements for the integration target, suggesting that the targeted DNA might assume specific secondary structures or protein associations. Here, we report that S1 SINE elements in the genomes of Brassica show an interesting preference for matrix attachment regions (MARs). Ten cloned genomic regions were tested for their ability to bind the nuclear matrix both before and after a SINE integration event. Eight of the genomic regions targeted by S1 display strong affinity for the nuclear matrix, while two show weaker binding. The SINE S1 did not display any matrix-binding capacity on its own in either non-methylated or methylated forms. In vivo, an integrated S1 is methylated while the surrounding genomic regions may remain undermethylated or undergo methylation. However, tested genomic regions containing methylated S1, with or without methylated flanking genomic sequences, were found to vary in their ability to bind the matrix in vitro. These results suggest a possible molecular basis for a preferential targeting of SINEs to MARs and a possible impact of the integration events upon gene and genome function.

Binding, Competitive↗

Molecular structure and regulatory potential of a T-DNA integration site in petunia.

The genomic structure surrounding a T-DNA integration site in a transgenic petunia plant, which shows deregulation of a root-specific promoter, was investigated. We have already demonstrated that T-DNA integration in this transformant (P13) had occurred close to a scaffold/matrix attachment region (S/MAR). A major question regarding the observed promoter leakiness was whether the T-DNA had integrated into the centre or at the border of the Petun-SAR and whether other regulatory elements are located within this genomic region. While small rearrangements were shown to occur during T-DNA integration in agreement with other reports, we find indications of the presence of a SINE retroposon--an apparent landmark for recombinogenic targets--at the integration site. Binding assays to both plant and animal nuclear scaffolds, supported by biomathematical analyses, reveal that the T-DNA is definitely located at the border of a strong S/MAR, which is in agreement with current models on the structure of integration sites. These results, together with a developmentally regulated leaf-specific enhancer effect of the Petun-SAR on gene expression in transgenic tobacco plants, indicate that the Petun-SAR demarcates the right border of a chromatin domain with genes predominantly active in leaves.

Agrobacterium tumefaciens↗

Sensitivity of indentation testing to step-off edges and interface integrity in cartilage repair.

Step-off edges and tissue interfaces are prevalent in cartilage injury such as after intra-articular fracture and reduction, and in focal defects and surgical repair procedures such as osteochondral graft implantation. It would be useful to assess the function of injured or donor tissues near such step-off edges and the extent of integration at material interfaces. The objective of this study was to determine if indentation testing is sensitive to the presence of step-off edges and the integrity of material interfaces, in both in vitro simulated repair samples of bovine cartilage defect filled with fibrin matrix, and in vivo biological repair samples from a goat animal model. Indentation stiffness decreased at locations approaching a step-off edge, a lacerated interface, or an integrated interface in which the distal tissue was relatively soft. The indentation stiffness increased or remained constant when the site of indentation approached an integrated interface in which the distal tissue was relatively stiff or similar in stiffness to the tissue being tested. These results indicate that indentation testing is sensitive to step-off edges and interface integrity, and may be useful for assessing cartilage injury and for following the progression of tissue integration after surgical treatments.

Animals↗

Edible oil crops and their integration with the major cereals in North Shewa and South Welo, Central Highlands of Ethiopia: an ethnobotanical perspective.

A total of 1050 sorghum (Sorghum bicolor) and tef (Eragrostis tef) fields distributed in six study sites within north Shewa and south Welo (Central Highlands of Ethiopia) were systematically surveyed to examine the status of integration of edible oil crops into the cereal-based farming system. Farmers' criteria for, and perception on, integration between edible oil crops and the major cereals at field level as well as integration between the cereal grains and the oilseeds in food making at home were studied and analyzed based on formal semi-structured interview and informal discussion with local expert farmers as key informants. Farmers' traditional space optimization technique has been instrumental in rightly fitting edible oil crops (as intercrops and border crops) into the cereal-based system. Six species of edible oil crops are integrated (70.3%) in various proportions in fields of sorghum and tef. At least one oil crop was significantly intercropped and/or border cropped in sorghum fields. Noog (Guizotia abyssinica) and sesame (Sesamum indicum) were the most important edible oil crops of the study area having strong integration with sorghum both at field and home level. On average, noog was more frequently intercropped with sorghum (8.3%) than with tef (4.5%), while it was more frequently border cropped with tef (32.4%) than with sorghum (19%). Sorghum was more frequently inter/border cropped with sesame (39/2%) than with tef (10.1/0.5%). The stronger the integration of a given oilseed with sorghum-based foods, the higher the companionship between sorghum fields and the oil crop in the landscape. This cultural practice by farmers has positive contributions to on-farm conservation of oil crops along with tef and specific sorghum landraces. The central theme of this paper therefore converges on the issue of on-farm in-situ agrobiodiversity conservation that was shaped by successive ancestral generations and passed on to the present.

Crops, Agricultural↗

Visual form integration and discontinuity detection.

Two experiments were conducted to determine if discontinuity detection limits the visual integration of two stimuli presented successively. If this is the case, then presenting two dim stimuli should permit better integration than presenting two bright stimuli. In Experiment 1, five observers named the position of the missing dot in a 5 x 5 dot matrix. Twelve randomly selected dots were presented in each of two stimulus presentations. The stimuli varied in intensity in a ratio of 15:1 and stimulus onset asynchrony (SOA) varied between 50 msec and 80 msec. At the longer SOAs, presenting two bright stimuli led to a higher percentage of correct responses than presenting two dim stimuli. There was no difference between those conditions at the shorter SOAs. In Experiment 2, 12 naive observers rated the amount of integration of the same stimuli on a 7-point scale, under the same presentation conditions. Rated integration was higher with two dim stimuli than with two bright stimuli. These apparently contradictory results are explained in terms of Hawkins and Shulman's distinction between two kinds of visual persistence. It is concluded that subjects can respond to integration and discontinuity detection separately. No evidence was found that discontinuity detection limits the integration.

Discrimination Learning↗