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PubMed1995

ComEA, a Bacillus subtilis integral membrane protein required for genetic transformation, is needed for both DNA binding and transport.

The competence-related phenotypes of mutations in each of the four open reading frames associated with the comE locus of Bacillus subtilis are described. comEA and comEC are required for transformability, whereas the products of comEB and of the overlapping comER, which is transcribed in the reverse direction, are dispensable. Loss of the comEA product decreases the binding of DNA to the competent cell surface and the internalization of DNA, in addition to exhibiting a profound effect on transformability. The comEC product is required for internalization but is dispensable for DNA binding. ComEA is shown to be an integral membrane protein, as predicted from hydropathy analysis, with its C-terminal domain outside the cytoplasmic membrane. This C-terminal domain possesses a sequence with similarity to those of several proteins known to be involved in nucleic acid transactions including UvrC and a human protein that binds to the replication origin of the Epstein-Barr virus.

Amino Acid Sequence↗
PubMed1992

Control of synthesis and secretion of ovarian oxytocin in ruminants.

Increased expression of the oxytocin gene of ruminants is associated with the process of luteinization both in vivo and in vitro. Cell culture studies and measurements of mRNA in luteal extracts have confirmed that the gene is switched on in the preovulatory follicle about 24 h before ovulation, at the time of the gonadotrophin surge. It is downregulated again equally rapidly after ovulation, so that by day 2 of the cycle the capacity of the luteal cells to make oxytocin has already been greatly reduced. A number of factors can increase oxytocin production by luteinizing granulosa cells. They include oestradiol and compounds such as gonadotrophins and catecholamines which are known to act by increasing intracellular concentrations of adenylyl cyclase. However, all of these factors are ineffective if the follicle is collected too early, suggesting that an initial maturation step is necessary to develop responsiveness. Analysis of the promoter region of the bovine oxytocin gene has indicated that neither oestradiol nor cAMP can directly initiate activation; instead regulation appears to occur via a COUP factor binding site. Additional transacting nuclear proteins may therefore be required to act as intermediaries. The same factors that initially stimulate oxytocin production switch to inhibiting production shortly after ovulation, leading to downregulation of the gene. After translation of oxytocin mRNA during the luteal phase, oxytocin precursor is packaged into secretory granules in the large luteal cells. Processing involves a series of enzymatic steps, culminating in amidation to produce oxytocin. Cultured cells may secrete intermediate forms of partially processed peptide, but it is not known if this also occurs in vivo. Oxytocin release from the cell involves granule exocytosis which is probably triggered by an increase in intracellular calcium. During luteolysis this is regulated by the release of prostaglandin F2 alpha (PGF2 alpha) from the uterus, although additional factors may also contribute. Neither PGF2 alpha nor catecholamines appear to be prime regulators of luteal oxytocin release during the early and mid-luteal phases of the cycle and it remains to be determined how secretion is controlled at this time.

Animals↗
PubMed1990

Isolation and characterization of cDNA encoding a human nuclear antigen predominantly recognized by autoantibodies from patients with primary biliary cirrhosis.

Autoantibodies to a novel nuclear Ag, Sp100, have recently been described that recognize a nuclear protein with an apparent molecular mass of 95 to 100 kDa and a dot-like distribution within cell nuclei. By immunoscreening of a lambda gt11 cDNA expression library derived from HeLa cells with an anti-Sp100 autoimmune serum a 0.7-kb cDNA (Sp26) coding for a fragment of Sp100 was isolated. Expression of this cDNA and use of the recombinant protein in ELISA revealed that the fragment carries major Sp100 autoepitopes and that anti-Sp100 autoantibodies predominantly occur in patients suffering from primary biliary cirrhosis (50/184). The Sp26 cDNA was used as hybridization probe for isolation of longer cDNA from human liver- and placenta-derived lambda gt10 cDNA libraries. Overlapping fragments were assembled to generate a full length cDNA coding for a protein with a molecular mass of 53 kDa and an isoelectric point of 4.7. The Sp100 autoantigen expressed in vitro from this cDNA and authenticated by a capture immunoblot assay, comigrated in SDS-PAGE with the authentic HeLa autoantigen of 95 to 100 kDa and thus showed an aberrant electrophoretic mobility. Computer based protein sequence analysis of the Sp100 autoantigen revealed regions of striking sequence similarities to the alpha 1 and alpha 2 domains of various human and non-human MHC class I Ag and to several transacting transcriptional regulatory proteins.

Amino Acid Sequence↗
PubMed1993

Basic mechanisms of cardiac gene expression.

Although the physiological properties of the myocardium and their dynamic character have been the focus of intense research during the past three decades, the biochemical and molecular correlates underlying cardiac development and performance have, until recently, remained poorly understood. The development of modern biology has provided the necessary tools to undertake the study of the mechanisms involved in cardiac development and to understand the basis for important clinical and experimental problems in cardiovascular physiology. Most of the gene encoding contractile proteins have been cloned and characterized. The availability of molecular probes and the ability to introduce genes into individual cell types and tissues of living animals are the most important breakthroughs of molecular and cell biology. This permits not only analysis of basic mechanisms of gene expression but has also significant practical applications for gene therapy. It is now possible to analyse the role of different regulatory gene sequences and identify their corresponding transactive factors. In addition direct gene injection makes it possible to study gene expression in a natural context, under conditions that are physiologically relevant and controllable.

Animals↗
PubMed2006

Isolation and minimization of head motion-induced signal variations in fMRI data using independent component analysis.

Task-related head movement during acquisition of fMRI data represents a serious confound for both motion correction and estimates of task-related activation. Cost functions implemented in most conventional motion-correction algorithms compare two volumes for similarity but fail to account for signal variability that is not due to motion (e.g., brain activation). We therefore recently proposed the theoretical basis for a novel method for fMRI motion correction, termed motion-corrected independent component analysis (MCICA), that allows for brain activation present in an fMRI time-series to be implicitly modeled and mitigates motion-induced signal changes without having to directly estimate the motion parameters (Liao et al., IEEE Transactions on Medical Imaging 2005;25:29-44). To explore the effects of non-movement-related signal changes on registration error, we performed several previously proposed test simulations (Freire et al., IEEE Transactions on Medical Imaging 2002;21:470-484) to evaluate the performance of MCICA and compare it with the conventional square-of-difference-based measures such as LS-SPM and LS-AIR. We demonstrate that for both simulated data and real fMRI images, the proposed MCICA method performs favorably. Specifically, in simulations MCICA was more robust to the addition of simulated activation, and did not lead to the detection of false activations after correction for simulated task-correlated motion. With actual data from a motor fMRI experiment, the time course of the derived continually task-related ICA component became more correlated with the underlying behavioral task after preprocessing with MCICA compared to other methods, and the associated activation map was more clustered in the primary motor and supplementary motor cortices without spurious activation at the brain edge. We conclude that assessing the statistical properties of a motion-corrupted volume in relation to other volumes in the series, as is done with MCICA, is an accurate means of differentiating between motion-induced signal changes and other sources of variability in fMRI data.

Algorithms↗
PubMed1998

Infant abuse and neglect in monkeys--a discussion of definitions, epidemiology, etiology, and implications for child maltreatment: reply to Cicchetti (1998) and Mason (1998)

In response to D. Cicchetti's (1998) and W. A. Mason's (1998) commentaries, this article scrutinizes infant maltreatment in monkeys and its public health implications. Studies of infant abuse and neglect in monkeys have used operational definitions based on (a) adult behavior or (b) adult behavior and infant outcome (depending on data available for analysis). Direct comparisons between the incidence of maltreatment in monkey and human populations can be only tentative as a result of differences in operational definitions of maltreatment. A simplified version of the ecological-transactional model of maltreatment can be used in research with nonhuman primates, and different species can be used to model different aspects of the human phenomenon. Although abuse and neglect take different forms in animals and humans, research with animal models can make an important contribution to elucidating the adaptive function, if any, of child maltreatment and the proximate mechanisms underlying its occurrence.

Animals↗
PubMed2006

Small company mergers--good for whom?

An analysis of 105 mergers among small UK biotech companies over a 10-year period shows that the improvement of shareholder positions, rather than product pipelines or business opportunities, is the main motivation for such transactions.

Biotechnology↗
PubMed2004

Implementing the HL7v3 standard in Croatian primary healthcare domain.

The mission of HL7 Inc. is to provide standards for the exchange, management and integration of data that supports clinical patient care and the management, delivery and evaluation of healthcare services. The scope of this work includes the specifications of flexible, cost-effective approaches, standards, guidelines, methodologies, and related services for interoperability between healthcare information systems. In the field of medical information technologies, HL7 provides the world's most advanced information standards. Versions 1 and 2 of the HL7 standard have on the one hand solved many issues, but on the other demonstrated the size and complexity of the health information sharing problem. As the solution, a complete new methodology has been adopted, which is being encompassed in version 3 recommendations. This approach standardizes the Reference Information Model (RIM), which is the source of all domain models and message structures. Message design is now defined in detail, enabling interoperability between loosely-coupled systems that are designed by different vendors and deployed in various environments. At the start of the Primary Healthcare Information System project, we have decided to go directly to HL7v3. Implementing the HL7v3 standard in healthcare applications represents a challenging task. By using standardized refinement and localization methods we were able to define information models for Croatian primary healthcare domain. The scope of our work includes clinical, financial and administrative data management, where in some cases we were compelled to introduce new HL7v3-compliant models. All of the HL7v3 transactions are digitally signed, using the W3C XML Digital Signature standard.

Croatia↗
PubMed2002

[Occupational stress and health in a sample of chilean executives].

BACKGROUND: Most individual and organizational costs related to job stress are due to preventable health problems. To study the impact of occupational stress in Chile, an instrument that evaluates the different variables involved in the stress process is required. AIM: To study the effects of work stressors and psychological variables on health among Chileans managers. To study reliability and validity of the Spanish version of the Occupational Stress Indicator (OSI-2). SUBJECTS AND METHODS: A translated version of the OSI-2 was applied to a sample of 264 white-collar managers. RESULTS: A high frequency of symptoms was reported by our sample. Back pain was the most frequently reported symptom in managers with high levels of stress. Regression analysis showed that, together, work stressors and psychological characteristics significantly explained self-perceived mental (27.6%) and physical (22.9%) health variance. Responsibilities of the executive role were the stressors most highly associated with poor stress outcomes. The psychological variables most strongly correlated with mental and physical health were Problem-focused Coping and the subscale Impatience of Type A Behavior. The reliability and validity of the OSI-2, Spanish version, were reasonably high. CONCLUSIONS: The transactional model of work stress was confirmed in a sample of Chilean managers evaluated with the Spanish version of the OSI-2.

Administrative Personnel↗
PubMed1990

A method of pharmacoepidemiologic analysis that uses computerized Medicaid.

A method of pharmacoepidemiologic data analysis that utilizes computerized Medicaid data is presented. A cohort design in which Medicaid enrollees receiving drugs that are normally used to treat similar underlying conditions is described. A period of time in which Medicaid service transactions are evident is required before an individual is eligible for selection into a cohort. Selection of study subjects and descriptions of cohorts are based on Medicaid service histories occurring during the preliminary, prerequisite period. Time at risk is considered to begin after a prescription for a study drug is dispensed and continues until either a refill is dispensed, a prescription for an alternative drug within the same therapeutic class is dispensed, or a predetermined number of days has passed. Subjects are followed forward in time and relevant health care transactions that are suggestive of suspected adverse drug reactions are noted. Incidence densities associated with sequentially ranked prescriptions within sequential courses of therapy are compared. Methods to increase the accuracy of case ascertainment are briefly discussed. Separate validation studies may be used to evaluate the validity of computerized case ascertainment methods and to compensate for misclassification of outcome. The proposed method is intended to provide timely estimates of risk for selected outcomes. For outcomes that cannot be accurately ascertained from computerized data, this method may be useful in determining the feasibility of more customized studies.

Age Factors↗
PubMed2000

Cell biology of cytochrome P-450 in the liver.

Cytochromes P-450 (P-450) are members of a multigene superfamily of hemoproteins consisting the microsomal monooxygenase system with NADPH P-450 reductase (reductase) and/or reducing equivalents. Expression of many P-450 isoforms in hepatocytes is shown to be regulated at the level of transcription through interaction between cis-acting elements in the genes and DNA-binding (transacting) factors. Some isoforms of the CYP1A, 2B, 2E, and 3A subfamilies are regulated at the posttranscriptional level. For the topology of P-450 and reductase molecules in ER membrane of hepatocytes, models from stopped flow analysis and electron spin resonance are proposed. The densities of total P-450 and reductase molecules are revealed to be high enough to support the cluster model, suggesting that about ten P-450 molecules form an aggregate and surround one reductase molecule, and therefore the two enzymes form large micelles. ER proliferation after PB administration, which had been correlated with increase in P-450 level, is shown to be probably independent of the increase in P-450 level. There are considerable discrepancies among results reported on sublobular expression of various P-450 isoforms. Causes of the discrepancies are likely to be differences in experimental conditions of histochemical detection carried out and/or in species, strain, and/or sex.

Animals↗
PubMed1999

Sites and mechanisms of antibiotic-induced neuromuscular block: a pharmacological analysis using quantal content, voltage clamped end-plate currents and single channel analysis.

Since the original observation of Vital Brazil and Corrado (1957) concerning the antibiotic induced neuromuscular block produced by streptomycin, there has been considerable interest in the mechanisms responsible for not only neuromuscular block but also the effects of antibiotics on different systems. We used the voltage clamped end-plate of transacted skeletal muscle to examine the concentration-dependent actions of several groups of antibiotics. The aminoglycoside antibiotics, neomycin and streptomycin, were both more effective at reducing quantal release of acetylcholine (ACh) than interacting with the postjunctional ACh receptor-channel complex. Neomycin was approximately 10 X more potent prejunctionally than streptomycin and the prejunctional effects of each antibiotic were reversed competitively by raising extracellular calcium. Both neomycin and streptomycin also had postjunctional actions at higher concentrations. Neomycin interacted with the open state of the ACh receptor ion channel complex while streptomycin blocks the ACh receptor. The lincosamide antibiotics, lincomycin and clindamycin produced their neuromuscular block postjunctionally by interacting with the open state of the ACh-receptor channel complex. Clindamycin is approximately 20 X more effective at blocking the open channel than was lincomycin. Using cell attached patch clamp recordings in cultured rat myotubes, we demonstrated a lincosamide-induced block of open ion channels with clindamycin having a much slower unblocking rate than lincomycin. Using epimers of the lincosamides, we demonstrated that lipophilicity of the molecule, rather than stereochemical considerations, is important for open channel blockade affecting primarily the "off" rate of channel blocking. This mechanism appears important for not only the lincosamide antibiotics but also for the postjunctional actions of the aminoglycoside antibiotics, particularly neomycin.

Animals↗
PubMed2000

Job uncertainty and health status for nurses during restructuring of health care in Alberta.

The Alberta health care system experienced dramatic changes after provincial funding cuts to health care from 1993 to 1996. As a result, stressors for nurses increased. The question of whether job uncertainty, working conditions, cognitive appraisal, and coping strategies influence the health of registered nurses in a context of health care restructuring was examined. Lazarus and Folkman's Transactional Model of Stress was used as the conceptual framework. A total of 271 registered nurses employed in a large, urban, acute-care teaching hospital responded to a self-administered survey questionnaire. Using multiple regression analysis, depression and self-reported physical health were analyzed. The data suggest that the threat of being placed on recall, having a coworker bumped or laid off, and perceived job security were adversely related to physical health. High primary appraisal of threat was associated with high levels of depression and poor physical health. In addition, the findings suggest that various coping strategies had both buffering and exacerbating effects on physical health and depression.

Adult↗
PubMed1998

Pygmalion in the cradle: observer bias against cocaine-exposed infants.

This study examined whether the widely disseminated negative image of the "cocaine baby" would lead adults to perceive a videotaped unexposed infant more negatively simply because they had been told the infant was prenatally cocaine-exposed. Two hundred and forty-nine students from three state universities used a seven-point Likert scale to rate either an African-American or a white female infant on 20 bipolar adjective-pairs. As predicted, participants who observed a labeled infant rated her more negatively than did those for whom the infant had not been labeled as cocaine-exposed. The potentially negative consequences of this documented bias toward cocaine-exposed infants should both alert and concern professionals and researchers. If adults view the behavior of a nonexposed infant more negatively merely because they believe that the infant has been exposed, then parents (biological and adoptive), professionals, and researchers may view and respond to the behavior of infants who are cocaine-exposed more negatively. Transactional models of development suggest a potential for self-fulfilling prophecy.

Adult↗
PubMed2006

Responsivity to offspring's expression of emotion among childhood-onset depressed mothers.

This study examined responsivity of mothers with childhood-onset depression (COD) in relation to children's overt expression of positive and negative emotion. It was hypothesized that COD and control mothers would differ in contingent responsivity to their children's expression of both positivity and different types of negative emotionality. Using observations and maternal reports of their own emotional reactions, COD mothers were found to be less responsive to children's expression of distress. A Gender x Group interaction was also found with respect to observed maternal responsiveness to child positivity, such that COD mothers were more responsive to girls' than boys' expression of positivity. The results are discussed in reference to transactional models of early child psychopathology.

Adult↗
PubMed1995

Importance of the most proximal GC box for activity of the promoter of human thyroid hormone receptor beta 1.

We wish to localize the sequences required for transcriptional expression of the thyroid hormone receptor beta 1 (TR beta 1). Constitutive activity of the promoter of human thyroid hormone receptor beta 1 was assessed by transient transfection of deletion constructs attached to luciferase as reporter, into P19, GH3, HepG2, H19-7, and COS1 cells. A 40-base pair fragment of the beta 1 promoter including the TATA box induced minimal luciferase activity, which was considered basal activity. The activities of various lengths of the beta 1 promoter were estimated relative to the minimal promoter in five cell lines. The region between -130 and -40 was crucial for constitutive activity in all cell lines. Further deletion analysis in HepG2 cells showed that two regions mainly augmented the transcriptional activity of the minimal 40 base pair fragment. One region located at -115 to -93, which is highly GC-rich, included the most proximal of five putative GC boxes present in the whole 1325-base pair promoter. A second region contributing to expression of TR beta 1 in HepG2 cells is at -70 to -40. Mutation of the most proximal GC box strongly suppressed transactivity of the whole promoter in P19 and HepG2 cells. In contrast, mutations in the other GC boxes did not suppress transactivation in P19 cells and slightly suppress activation in HepG2 cells. In Schneider cells, which do not express Sp1, transactivity of the region distal to -40 is positively regulated by cotransfection with a vector expressing Sp1.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1984

Psychosocial metaphors of physical distress among MS patients.

This paper explores the dynamics between chronic illness and culture and analyzes how a person's expressions of chronic illness are culturally constituted. The analysis focuses on the ways physical symptoms among multiple sclerosis patients are expressed and experienced as psychosocial problems. The process of transforming a biological construct into a social one is discussed. These transformations are explored through the doctor/patient transaction and the solidification of psychosocial problems among MS patients during their interaction with their 'significant others' and then transferred to their larger social-cultural sphere.

Adult↗
PubMed1989

Developmentally regulated male-specific transfactor(s) enable in vitro transcription of a cloned alpha 2u-globulin gene.

Selected members of the rat alpha 2u-globulin gene family are expressed in several tissues, manifesting characteristic developmental and endocrine transcriptional control. Studies are now underway to identify the responsible cis sequences and transacting factors. We recently reported that the cloned rat alpha 2u-globulin 207 gene manifests tissue-specific androgen-dependent expression; it is expressed in livers of male, but not female, transgenic mice. In the present study a portion of this gene (-639 to +1395) is used as an in vitro template in the presence of nuclear extracts derived from hepatic nuclei of prepubescent and mature male and female rats. alpha-Amanitin-sensitive in vitro transcription of the alpha 2u 207 gene by extracts derived from mature male rats is highly active and is at least 13 times as rapid as that with preparations from mature female or immature animals. In contrast, the rate of transcription of a control template, the adenoviral late promoter, is the same with all of these nuclear extracts. S1 nuclease analysis and the size of the transcript indicate that transcription is initiated in vitro at the same site as it is in vivo and that it continues to the 3' terminus of the alpha 2u-globulin template. Thus, cis sequences are present in this gene fragment which are controlled by developmentally regulated male transacting factors, enabling selective transcription of this alpha 2u-globulin gene. Mixing experiments indicate that this transcriptional control is positive.

Alpha-Globulins↗