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PubMed2006

Characterization and comparison of raft-like membranes isolated by two different methods from rat submandibular gland cells.

Lipid rafts are defined as cholesterol and sphingolipid enriched domains in biological membranes. Their role in signalling and other cellular processes is widely accepted but the methodology used for their biochemical isolation and characterization remains controversial. Raft-like membranes from rat submandibular glands were isolated by two different protocols commonly described in the literature; one protocol was based on selective solubilization by Triton X-100 at low temperature and the other protocol consisted in extensive sonication. In both cases a low density vesicular fraction was obtained after ultracentrifugation in a sucrose density gradient. These fractions contained about 20% of total cholesterol but less than 8% of total proteins, and were more rigid than bulk membranes. Fatty acid analyses revealed a similar composition of raft-like membranes isolated by the two different methods, which was characterized by an enrichment in saturated fatty acids in detriment of polyunsaturated acids when compared with the whole cell membranes. Protein profile of detergent resistant membranes or raft-like membranes prepared by sonication was assessed by silver staining after SDS-PAGE and by MALDI-TOF. Both analyses provided evidence of a different protein composition of the Triton X-100 and sonication preparations. Immunoblot experiments revealed that raft-like membranes prepared by detergent extraction or sonication were free of Golgi apparatus or endoplasmic reticulum protein markers (beta-COP and calnexin, respectively) and that they were not substantially contaminated by transferrin receptor (a non-raft protein). While caveolin-1 was highly enriched in raft-like membranes prepared by the two methods, the P2X(7) receptor was enriched in raft-like membrane fractions prepared by sonication, but almost undetectable in the detergent resistant membranes. It can be concluded that both methods can be used to obtain raft-like membranes, but that detergent may affect protein interactions responsible for their association with different membrane domains.

Animals↗
PubMed2004

Differential Ca2+ and Sr2+ regulation of intracellular divalent cations release in ventricular myocytes.

The regulation of the Ca2+ -induced Ca2+ release (CICR) from intracellular stores is a critical step in the cardiac cycle. The inherent positive feedback of CICR should make it a self-regenerating process. It is accepted that CICR must be governed by some negative control, but its nature is still debated. We explore here the importance of the Ca2+ released from sarcoplasmic reticulum (SR) on the mechanisms that may control CICR. Specifically, we compared the effect of replacing Ca2+ with Sr2+ on intracellular Ca2+ signaling in intact cardiac myocytes as well as on the function of single ryanodine receptor (RyR) Ca2+ release channels in panar bilayers. In cells, both CICR and Sr2+ -induced Sr2+ release (SISR) were observed. Action potential induced Ca2+ -transients and spontaneous Ca2+ waves were considerably faster than their Sr2+ -mediated counterparts. However, the kinetics of Ca2+ and Sr2+ sparks was similar. At the single RyR channel level, the affinities of Ca2+ and Sr2+ activation were different but the affinities of Ca2+ and Sr2+ inactivation were similar. Fast Ca2+ and Sr2+ stimuli activated RyR channels equally fast but adaptation (a spontaneous slow transition back to steady-state activity levels) was not observed in the Sr2+ case. Together, these results suggest that regulation of the RyR channel by cytosolic Ca2+ is not involved in turning off the Ca2+ spark. In contrast, cytosolic Ca2+ is important in the propagation global Ca2+ release events and in this regard single RyR channel sensitivity to cytosolic Ca2+ activation, not low-affinity cytosolic Ca2+ inactivation, is a key factor. This suggests that the kinetics of local and global RyR-mediated Ca2+ release signals are affected in a distinct way by different divalent cations in cardiac muscle cells.

Action Potentials↗
PubMed2006

Road de-icing salt as a potential constraint on urban growth in the Greater Toronto Area, Canada.

North America's fifth most populated municipality--the Greater Toronto Area (GTA)--is undergoing rapid urban development with serious questions being raised regarding the long-term impacts of urban growth on the quality and quantity of ground and surface water. Degradation of groundwater quality by NaCl de-icing salt is the primary concern since there are no cost effective alternatives to NaCl de-icing salt and there is little evidence that salt loadings to the subsurface can be significantly reduced. In 2001, the issue acquired a new sense of urgency when de-icing chemicals containing inorganic chloride salts (with or without ferrocyanide de-caking agents) were designated as toxic under the Canadian Environmental Protection Act. To heighten concerns, future growth in the GTA will inevitably take place in areas where groundwater is regularly used for potable supply. Studies using groundwater flow and transport models show that significant deterioration of groundwater quality can be expected in shallow aquifers as a result of urban development with chloride concentrations approaching the drinking water quality standard of 250 mg/l. Results demonstrate that urban planning needs a fresh approach that explicitly includes groundwater protection and aquifer management in the decision-making process, clearly defines acceptable environmental performance standards and makes greater use of groundwater models to evaluate alternative urban designs.

Canada↗
PubMed2004

Consistently high plasma high-density lipoprotein-cholesterol levels in children in Spain, a country with low cardiovascular mortality.

Coronary heart disease (CHD) mortality is relatively low in Spain compared with other developed countries and has remained low despite an apparent increase in mean plasma cholesterol concentration in adults over the last several years. It is accepted that pathologic processes related to arteriosclerosis development begin in childhood and seem to be related to the presence of cardiovascular risk factors at this age. High-density lipoprotein-cholesterol (HDL-C) levels in children have been inversely correlated with the incidence of coronary heart disease in the different countries studied. Childhood plasma lipoprotein profile might contribute to the low coronary heart disease mortality in Spain. Thus, we analyzed data on lipid levels over time in schoolchildren in Spain in the last decade. Plasma lipid levels were analyzed in prepuberal children (6 to 8 years) in 3 school-based surveys performed by our group in Madrid in 1987, 1993, and 1999. A significant increase in plasma total cholesterol (P < .05) and low-density lipoprotein-cholesterol (LDL-C) (P < .01) levels in prepuberal children was observed over the last decade. However, the mean concentration of plasma HDL-C remained stable and very high. These high levels of plasma HDL-C in Spanish school children may help to explain why the coronary heart disease mortality rate in Spain is low compared with that in other developed countries.

Apolipoproteins↗
PubMed2004

Induced depressive behavior impairs learning and memory in rats.

While it is generally accepted that cognitive processes such as learning and memory are affected by emotion, the impact of depression on learning and memory has rarely been directly studied in experimental animals. Effects of induced depressive behavior on learning and memory were determined in rats, using an open space swim test, a novel animal model of depressive behavior that is developed recently in our laboratory. The model indexes searching activity of the animals, with the induced depressive immobility behavior showing specific sensitivity to three major prototypic classes of antidepressants and a selective serotonin reuptake inhibitor. The induced depressive behavior in rats showed a delayed response to chronic antidepressant treatment and had a lasting effect on the ability of rats to learn and recall the learned experience. It impaired the subsequent ability of rats to learn and recall both a spatial water maze task and a multi-trial passive avoidance task. These impairments were all sensitive to antidepressant therapeutics, but not to buspirone, an anxiolytic. By way of contrast, the ability of the rats to sense and move to a visible platform and to escape from an unconditioned shock stimulus was neither impaired by inducing the depressive behavior nor altered by the drug treatment, suggesting that non-specific changes in sensorimotor ability were not involved. These impairments of learning and memory indicate that the depressive behavior-induced deficits show generalizability and are not context-limited. This animal model of depressive behavior shows promising potential as a screen for novel antidepressive therapeutics and as a disease model for revealing network/cellular/molecular mechanisms in the pathophysiology of depression and depression-induced cognitive deficits.

Animals↗
PubMed2003

Historical review: the discovery of 'giant' RNA and RNA processing: 40 years of enigma.

RNA processing is a primordial paradigm of gene expression. Iconoclastic when discovered, after 40 years there is still no general rationale for this apparent 'wasting' of up to 90% of RNA transcripts. This article tells the story of the discovery of RNA in the laboratory of J.E. Darnell. The discovery of 'giant' RNA and its conversion into rRNA revealed the phenomenon of RNA processing and pre-rRNA. Genuine mRNA was also identified, but the majority of DNA-like nuclear RNA was also found to be giant and unstable. In spite of early evidence, pre-mRNA processing was only accepted in 1977 when the discovery of gene fragmentation in DNA made it obvious.

Animals↗
PubMed2002

Targeting ergonomics interventions--learning from health promotion.

Ergonomics often involves encouraging people to accept change, a process affected by their knowledge, attitudes and beliefs. This paper argues that ergonomics projects are likely to have greater success where formal account is taken of these characteristics, and interventions are tailored accordingly. Furthermore, it is suggested that ergonomics can usefully draw upon behaviour change models developed by those concerned with health promotion in the community. One influential model is Prochaska and DiClemente's (Psychotherapy: Theory, Research and Practice, 19, 276-288, 1982) stage of change framework, which describes recipients of health information as progressing through six stages towards stable healthy behaviour: precontemplation, contemplation, preparation, action, maintenance and relapse. This recognises the dynamic nature of behaviour change and that, both within and between individuals, responses may vary from failing to acknowledge existence of a health risk, to where something has already been done about it. Proponents of the model contend that interventions should seek to identify recipients' stage of change and target information and advice accordingly. It is proposed that Prochaska and DiClemente's approach might usefully inform ergonomics activities, particularly when advising with respect to health and safety (e.g. manual handling, upper limb disorders, fall risk, plant safety, etc.) but perhaps also where dealing with other ergonomics considerations (i.e. comfort and performance).

Ergonomics↗
PubMed1997

Autoimmunity and reproduction.

OBJECTIVE: To review the association between autoimmunity and reproductive failure. DESIGN: A MEDLINE search done from 1965 to 1996. More than 300 original and review articles were evaluated, from which the most relevant were selected. RESULT(S): Autoimmune processes now are accepted widely as one of the possible mechanisms of many human diseases. The presence of autoimmune disorders has been associated repeatedly with reproductive failure. On the other hand, reproductive failure may be the first manifestation of autoimmune disorders. CONCLUSION(S): When abnormal autoantibody levels are present in women with reproductive failure, the reproductive failure alone should be considered as one of the possible clinical expressions of autoimmune disorders. Two relevant questions of whether these patients should be treated for autoimmunity remain unsolved. A prospective, placebo-controlled trial is necessary to evaluate the importance of any treatment.

Abortion, Habitual↗
PubMed1988

Pediatric Symptom Checklist: screening school-age children for psychosocial dysfunction.

The Pediatric Symptom Checklist (PSC) is a 35-item screening questionnaire that is completed by parents and designed to help pediatricians in outpatient practice identify school-age children with difficulties in psychosocial functioning. The current study assessed the validity of the PSC by screening 300 children in two pediatric practices, a middle-class group practice and an urban health maintenance organization. Validity was established by comparing the results of PSC screening of 48 children with in-depth interview assessments and pediatricians' ratings. Results indicate that the PSC has a specificity of 0.68 and a sensitivity of 0.95. The screening process was well accepted by parents and pediatricians. Several children whose pediatricians' ratings had indicated adequate functioning were identified by the PSC as having substantial psychosocial dysfunction and requiring further evaluation.

Child↗
PubMed2002

The sequestration switch: removing industrial CO2 by direct ocean absorption.

This review paper considers direct injection of industrial CO2 emissions into the mid-water oceanic column below 500 m depth. Such a process is a potential candidate for switching atmospheric carbon emissions directly to long term sequestration, thereby relieving the intermediate atmospheric burden. Given sufficient research justification, the argument is that harmful impact in both the Atmosphere and the biologically rich upper marine layer could be reduced. The paper aims to estimate the role that active intervention, through direct ocean CO2 storage, could play and to outline further research and assessment for the strategy to be a viable option for climate change mitigation. The attractiveness of direct ocean injection lies in its bypassing of the Atmosphere and upper marine region, its relative permanence, its practicability using existing technologies and its quantification. The difficulties relate to the uncertainty of some fundamental scientific issues, such as plume dynamics, lowered pH of the exposed waters and associated ecological impact, the significant energy penalty associated with the necessary engineering plant and the uncertain costs. Moreover, there are considerable uncertainties regarding related international marine law. Development of the process would require acceptance of the evidence for climate change, strict requirements for large industrial consumers of fossil fuel to reduce CO2 emissions into the Atmosphere and scientific evidence for the overall beneficial impact of ocean sequestration.

Absorption↗
PubMed1999

Modulation of DNA damage and DNA repair in chromatin.

DNA is packaged in the highly compact and dynamic structure of chromatin in eukaryotic cells. It is generally accepted that DNA processing events in the nucleus, such as transcription, replication, recombination, and repair, are restricted by this packaging. For some processes (e.g., transcription), the chromatin fiber is "preset" in a more open structure to allow access of proteins to specific regions of DNA within this structural hierarchy. These regions contain modified nucleosomes that accommodate a less compact state of chromatin and allow access to specific regions of DNA. DNA repair proteins, however, must access DNA lesions in all structural domains of chromatin after sudden insult to the genome. Damaged DNA must be recognized, removed, and replaced by repair enzymes at all levels of chromatin packaging. Therefore, the modulation of DNA damage and its repair in chromatin is crucial to our understanding of the fate of potential mutagenic and carcinogenic lesions in DNA. In this review, we discuss the modulation of DNA damage and DNA repair by chromatin structure, and the modulation of chromatin structure by these events.

Animals↗
PubMed1997

A quantitative assessment of the risk of transmission of foot-and-mouth disease, bluetongue and vesicular stomatitis by embryo transfer in cattle.

This paper addresses the risks involved when bovine embryos are moved internationally and, specifically, the possibilities of transmitting foot-and-mouth disease, bluetongue and vesicular stomatitis by embryos originating from an area in South America. The risk scenario pathway was divided into three phases for analysis. The first phase dealt with the potential for embryo contamination which depends on the disease situation in the exporting country and/or region, the health status of the herds and the donor cows from which the embryos are collected, and the pathogenetic characteristics of the specified disease agent. The second phase covers risk mitigation by use of internationally accepted standards for processing of embryos, and the third phase encompassed the risk reductions resulting from post-collection surveillance of the donors and donor herds, and also from testing of embryo-collection (flushing) fluids for the disease agent. Quantitative risk analysis showed that under the circumstances specified in the paper, the risk of transmission of foot-and-mouth disease and vesicular stomatitis by embryos would be likely to be less than 1 in 100 billion (10(-11.0)) and 1 in 100 million (10(-8.0)), respectively. The values for bluetongue were 1 in 30,000 (10(-4.2)) when embryos were collected in the vector season and 1 in 1 million (10(-6.0)) in the season with low vector activity. These risk values were influenced by the incidence of each disease in the area of origin and the ease with which clinical signs can be recognised. Competent embryo processing according to procedures recommended by the International Embryo Transfer Society were also of great importance. The analysis showed that the reasons for the low levels of risk of transmission differed for each of the three diseases. In the case of bluetongue, vector ecology was of major importance.

Animals↗
PubMed2002

Entering the well-guarded fortress: alternative practitioners in hospital settings.

There is a growing evidence that alternative health care practitioners and physicians are working together in collaborative patterns. The paper examines these collaborative patterns in hospital settings in Israel. On the theoretical level, the specific issues relate to theories concerning relationships between dominant institutional structures which enjoy the benefits of epistemological legitimacy as well as extensive, supportive social structures and groups of non-conformists who seek to attain many of the same goals by utilizing different methods based on other epistemologies. In the most general sense, the issues involved concern processes of accommodation and social change. Data were collected by means of semi-structured, qualitative interviews in four general hospitals in Jerusalem during 2000. Nineteen persons were interviewed including 10 alternative practitioners working in a variety of fields and nine biomedical practitioners who worked with them (six physicians and three nurses). Interviews focused on background and training, reasons for entry into the hospital, length of practice, status in the hospital system, mode of remuneration, content of work, modes of interaction with others in the hospital and problems encountered. The findings suggest a dual process of simultaneous acceptance and marginalization of alternative practitioners. While small numbers of alternative practitioners were found to be practicing in a wide variety of hospital departments and in a broad spectrum of specialties, they were in no way accepted as regular staff members and their marginality was made clear by a variety of visible structural, symbolic and geographical cues. There is a division of labour expressed by focusing on the biomedical practitioners on the diagnosis and treatment of specific disease entities, while the alternative practitioners work in the illness context, concentrating of feelings and affective states involving the alleviation of pain, suffering and efforts to improve the quality of life.

Ancillary Services, Hospital↗
PubMed2001

Tissue-specific expression of human ERalpha and ERbeta in the male.

The important role of estrogens in women in physiological and pathological processes is well accepted, but recently it has become evident that estrogens are also important in male physiology, in particular, within bone metabolism and reproduction. Consequently, it is necessary to identify and to characterize the molecular mechanisms of estrogen action in order to evaluate how the pleiotropic effects of estrogens are mediated in a variety of tissues. We have recently shown that human estrogen receptor alpha (ERalpha) mRNA is transcribed from at least six different promoters (1A-1F). Transcription of ERalpha in bone is exclusively dependent on the F-promoter. To study the regulation of ER expression in this tissue, we examined 1 kbp of the F-promoter region of human ERalpha, which is located more than 70 kbp upstream of the transcription start site of the ERalpha gene. Transient transfection experiments demonstrated a basal activity from the F-promoter, which was further increased when ERalpha was cotransfected. We have shown recently that the F-promoter can give rise to at least two ERalpha isoforms in bone. On the contrary, ERbeta expression in primary osteoblasts is extremely low, indicating that this ER isoform plays only a minor role in these cells. In contrast to bone, we have demonstrated that both ERalpha and ERbeta transcripts are readily detected in testis. Here, we report that besides ERalpha, ERbeta transcripts can give rise to two protein isoforms and that this complex situation could have important functional consequences for the signalling of estrogens and their analogs.

Alternative Splicing↗
PubMed2001

Ultrasonic depolymerization of aqueous polyvinyl alcohol.

Ultrasonication has proved to be a highly advantageous method for depolymerizing macromolecules because it reduces their molecular weight simply by splitting the most susceptible chemical bond without causing any changes in the chemical nature of the polymer. Most of the effects involved in controlling molecular weight can be attributed to the large shear gradients and shock waves generated around collapsing cavitation bubbles. In general, for any polymer degradation process to become acceptable to industry, it is necessary to be able to specify the sonication conditions which lead to a particular relative molar mass distribution. This necessitates the identification of the appropriate irradiation power, temperature, concentration and irradiation time. According to the results of this study the reactors constructed worked well in depolymerization and it was possible to degrade aqueous polyvinyl alcohol (PVA) polymer with ultrasound. The most extensive degradation took place at the lowest frequency used in this study, i.e. 23 kHz, when the input power was above the cavitation threshold and at the lowest test concentration of PVA, i.e. 1% (w/w). Thus this study confirms the general assumption that the shear forces generated by the rapid motion of the solvent following cavitational collapse are responsible for the breakage of the chemical bonds within the polymer. The effect of polymer concentration can be interpreted in terms of the increase in viscosity with concentration, causing the molecules to become less mobile in solution and the velocity gradients around the collapsing bubbles to therefore become smaller.

Journal Article↗
PubMed2004

Ultrasonic depolymerization of aqueous carboxymethylcellulose.

Prolonged exposure of solutions of macromolecules to high-energy ultrasonic waves produces a permanent reduction in viscosity. However, the exact mechanism by which degradation occurs is still open to discussion. According to this study hydrodynamic forces played the primary role in degradation process. This study showed that there is an optimal carboxymethylcellulose (CMC) concentration to the most efficient degradation. Ultrasound degraded preferentially large CMC molecules and cleavage took place roughly at the centre of the CMC molecules. Degradation of CMC did not proceed below a certain molecular mass. During ultrasonic degradation the molecular mass distribution narrowed. For any polymer degradation process to become acceptable to industry, it is important to be able to specify the sonication conditions to produce a particular relative molecular mass distribution.

Journal Article↗
PubMed2003

African American grandmothers parenting AIDS orphans: concomitant grief and loss.

Ten urban African American grandmothers participated in a qualitative study using in-depth open-ended interviews to determine if and how the African American cultural experience informed the grieving process and the acceptance of the "off-time" parenting role. The findings do not support the premise that grief resolution requires the bereaved to relinquish all ties to the deceased. The participants maintained strong bonds with the deceased through the conscious decision to remember their unique qualities and through parenting their children, a tradition rooted in the history of the African American family. A spiritual relationship with God was a significant source of strength that enhanced their ability to parent while mourning.

Acquired Immunodeficiency Syndrome↗
PubMed1994

Pharmacokinetics and pharmacodynamics of MK-383, a selective non-peptide platelet glycoprotein-IIb/IIIa receptor antagonist, in healthy men.

MK-383 (L-tyrosine, N-(n-butylsulfonyl)-O-[4-butyl(4-piperidinyl)], monohydrochloride monohydrate) is a potent and specific platelet fibrinogen receptor antagonist that may be useful in preventing processes that lead to occlusive thrombus formation in the lumen of the blood vessel. Two placebo-controlled phase I trials were completed in 56 healthy volunteers to investigate the safety, tolerability, pharmacokinetics, and pharmacodynamics of MK-383 administered as 1- and 4-hour infusions in the presence and absence of aspirin. When administered to healthy male subjects by constant infusions up to 0.4 microgram/kg/min over 1 hour or up to 0.2 microgram/min over 4 hours, it provided a well-tolerated reversible means of inhibiting platelet function. At infusion rates of 0.25 and 0.15 microgram/kg/min for 1 and 4 hours, respectively, MK-383 extended baseline bleeding time by 2.0- to 2.5-fold and inhibited adenosine diphosphate (ADP)-induced platelet aggregation by at least 80%. The pharmacokinetics of MK-383 include a mean plasma clearance of 329 ml/min, steady-state volume of distribution of 76 L, and half-life of 1.6 hours. The percentage of dose excreted in the urine was 37%. Correlations between MK-383 plasma concentration (C) and inhibition of platelet aggregation were examined by fitting with a sigmoid maximum-effect model. The plasma concentration yielding 50% inhibition (C50) for MK-383 in healthy volunteers is approximately 13 ng/ml, with a Hill coefficient > 5. Based on a naive pooled analysis, an exponential empirical model best describes the MK-383 C-extension of template bleeding time (BTE) relationship. The model indicates that the MK-383 plasma concentration necessary to double BTE is approximately 30 ng/ml (i.e., 2.5-fold greater than the C50 for ADP-induced inhibition of platelet aggregation). The pharmacokinetics of MK-383 was unaffected by pretreatment with 325 mg aspirin 1 day before and 1 hour before infusion. Conversely, aspirin pretreatment reduced C50 and increased bleeding time extension, suggesting that aspirin may have an additive effect with respect to inhibition of platelet function. Based on the putative role of the fibrinogen receptor in thrombotic processes and an acceptable human pharmacokinetic-pharmacodynamic profile, MK-383 should be evaluated in patients with unstable angina.

Analysis of Variance↗