Effect of norgestrel on development of mouse pre-embryos

The effect of synthetic progestins found in oral contraceptives has potential implications for developing embryos in women who receive oral contraceptives during early pregnancy. We assessed the effect of the progestin norgestrel on the developing pre-embryo. B3C6F1 mice were given 5 IU PMSG followed by 5 IU hCG 48h later. Studies were performed on pre-embryos recovered and pooled at both 24h (Group A) and 48h (Group B) post hCG. At each time period, they were randomly assigned to control or norgestrel (4.0 ng/ml) treatment. In a third study, pre-embryos collected 24h post hCG were cultured in the absence or presence of 8.0, 80.0, or 800 ng/ml norgestrel. Cultures were performed in Ham's F-10 media with 10% fetal calf serum at 37 degrees C in an atmosphere of 5% CO2, 5% O2 and 90% N2 with 15-30 embryos per 1 ml of culture fluid. At 24h, 48h and 72h post recovery, cultures were viewed, the appearance of embryos noted, and number of blastomeres recorded. Compared to control groups, analysis demonstrated no significant difference in the rate of development of control and norgestrel pre-embryos in any group at any time period (24h, 48h, or 72h post recovery). We conclude that norgestrel at the dose tested has no acute adverse morphological effects on development of mouse pre-embryos. This observation has potential clinical implications with regard to inadvertent use of norgestrel-containing oral contraceptives during early days of pregnancy, as well as consideration of the mechanism of action of norgestrel-containing "morning after" pills.     

Assessment of Skin Barrier Function Using Transepidermal Water Loss: Effect of Age

To probe age-related changes in skin barrier function, transepidermal water loss (TEWL) rates have been measured in young (19–42 years) and old (69–85 years) subjects. TEWL was determined at ventral forearm skin sites, which had been occluded for 24 hr with polypropylene chambers. Baseline TEWL rates (J , which showed no dependence on age, were measured for each subject before and after the experiment. Following removal of the occlusive chamber, TEWL was monitored continuously from t = 0.5 min until its return to the baseline (preocclusion) level, which was typically in the range of 2–7 g/m²/hr. Initial TEWL rates (mean ± SD) were found to differ significantly between young (28.6 ± 7.5 g/m²/hr; n = 26) and old (36.9 ± 10.5 g/m²/hr; n = 18) subjects (P < 0.01). Relaxation of TEWL to J was significantly slower in the aged cohort, such that the characteristic time for diffusion of water in the stratum corneum was estimated to be (mean ± SD) 176 ± 59 min for the young subjects, compared to 360 ± 76 min for the old (P < 0.001.). Thus, the initial TEWL value following removal of occlusion is significantly greater, and the excessive stratum corneum hydration produced by occlusion is dissipated more slowly, in old skin than in young. A hypothesis to explain the slower relaxation of perturbed TEWL in old skin is proposed.     

Polymères thiocarboxyliques, 1. Synthèse et étude de la polymérisation radicalaire de monomères dithiocarboxyliques

The syntheses and free-radical polymerizations of methyl and carboxymethyl 4-vinyldithiobenzoates are described as well as various copolymers. Copolymerisation parameters were determined for the copolymerizations of the methyl dithioester with styrene and with methyl methacrylate.     

Morphology of Marburg virus NP-RNA

When Marburg virus (MBGV) nucleoprotein (NP) is expressed in insect cells, it binds to cellular RNA and forms NP-RNA complexes such as insect cell-expressed nucleoproteins from other nonsegmented negative-strand RNA viruses. Recombinant MBGV NP-RNA forms loose coils that resemble rabies virus N-RNA. MBGV NP monomers are rods that are spaced along the coil similar to the nucleoprotein monomers of the rabies virus N-RNA. High salt treatment induces tight coiling of the MBGV NP-RNA, again a characteristic observed for other nonsegmented negative-strand virus N-RNAs. Electron microscopy of fixed Marburg virus particles shows that the viral nucleocapsid has a smaller diameter than the free, recombinant NP-RNA. This difference in helical parameters could be caused by the interaction of other viral proteins with the NP-RNA. A similar but opposite phenomenon is observed for rhabdovirus nucleocapsids that are condensed by the viral matrix protein upon which they acquire a larger diameter. Finally, there appears to be an extensive and regular protein scaffold between the viral nucleocapsid and the membrane that seems not to exist in the other negative-strand RNA viruses.     

Improving HIV surveillance in Victoria: the role of the "detuned" enzyme immunoassay

Between 1999 and 2000, new diagnoses of HIV in Victoria (Australia) rose by 41%, from 140 to 197. In this time period, sera from new HIV diagnoses were tested using the Organon Teknika "detuned" enzyme immunoassay (EIA). We compared the results of the detuned EIA with incident infections defined by surveillance (on the basis of a previous negative or indeterminate HIV test and/or a seroconversion illness within the 12 months preceding HIV diagnosis). Of 317 specimens, 97 (31%) incident infections and 114 (36%) recent infections were detected using surveillance and detuned EIA, respectively. The detuned assay misclassified 11 cases with AIDS and 2 cases with CD4 counts < or = 200 micro3 (probable long-standing infections) as recent infections and was unable to identify 31 (32%) of 97 cases previously classified as incident cases by surveillance. The assay detected an extra 35 recent infections that were previously classified as nonincident by surveillance. By combining the detuned assay and surveillance, 132 (42%) incident infections were identified from 317 specimens, 36% more than surveillance alone. We recommend that a detuned assay or similar test become part of the routine strategy to identify incident infections in Victoria. Incident infections provide important information for targeting prevention strategies and the opportunity to interrupt ongoing viral transmission.     

The tyrosine-based YXXØ targeting motif of murine leukemia virus envelope glycoprotein affects pathogenesis

Retroviruses, such as human and simian immunodeficiency viruses (HIV and SIV), and murine leukemia viruses (MuLV), harbor a tyrosine-based motif in the intracytoplasmic domain of their envelope glycoprotein. This motif can act as an endocytosis signal or as a targeting signal, restricting viral budding at specific cell surface membrane domains. In the present study, proviral DNA of the ecotropic Cas-Br-E strain of MuLV was modified by substitution or deletion of the critical tyrosine residue. Mutant viruses lost basolateral targeting in polarized MDCK epithelial cells while expression level of the glycoprotein at the cell surface was not affected. This suggests that the tyrosine-based motif in MuLV does not act as an endocytosis signal. Only a small delay in the appearance of disease was observed in inoculated mice. In contrast, a striking change in the pathology was observed with enlarged thymus and lymph nodes in animals inoculated with mutant viruses.     

Fibrillar beta-amyloid peptide Aβ1–40 activates microglial proliferation via stimulating TNF-α release and H2O2 derived from NADPH oxidase: a cell culture study

Background  

Alzheimer's disease is characterized by the accumulation of neuritic plaques, containing activated microglia and β-amyloid peptides (Aβ). Fibrillar Aβ can activate microglia, resulting in production of toxic and inflammatory mediators like hydrogen peroxide, nitric oxide, and cytokines. We have recently found that microglial proliferation is regulated by hydrogen peroxide derived from NADPH oxidase. Thus, in this study, we investigated whether Aβ can stimulate microglial proliferation and cytokine production via activation of NADPH oxidase to produce hydrogen peroxide.     

Clinical information technology in hospitals: a comparison between the state of Iowa and two provinces in Canada

Despite the growing interest in adopting information technology (IT) in healthcare, the degree of technology sophistication varies among healthcare organizations. Changes in the health care sector and continuous pressure to improve the quality of care have driven the evolution of IT in hospitals. This paper provides an overview of clinical IT sophistication in a sample of U.S. hospitals, and compares clinical IT capacities in this sample with a sample of Canadian hospitals. The instrument used for the comparison measures three clinical dimensions of IT sophistication: functional sophistication, technological sophistication and integration level. Clinical areas that were considered include patient management, patient care activities and clinical support activities. The comparison between hospitals in Iowa and Canada shows differences in clinical IT sophistication between the two settings. Hospitals in Iowa appear to have more technologies but fewer computerized processes and integration of patient management activities. Technological sophistication however, was low in both samples. Our findings confirm the construct validity of the measurement instrument and show initial evidence of its generalizability. More initiatives using the instrument would lead to enhancement in IT assessment tools that can be used for evaluation of IT in relation to patient management and quality outcomes.     

Nociceptive stimulation activates locus coeruleus neurones projecting to the somatosensory thalamus in the rat

In the thalamus, noradrenergic output from the pontine nucleus locus coeruleus (LC) may actively shape the response properties of various sensory networks en route to the cortex. Little is known, however, about the involvement of ascending noradrenergic innervation of the somatosensory thalamus in the processing of nociceptive information. To address this question, we combined the study of Fos expression upon nociceptive tooth pulp stimulation in the anaesthetized rat, with the detection of retrogradely traced neurones from the somatosensory thalamus. Cell bodies labelled retrogradely from the left thalamus were observed on both sides of the LC, with an ipsilateral predominance ( n = 8). Electrical stimulation of the right incisor pulp ( n = 4) provoked a significantly stronger Fos expression (around twice) than sham surgery ( n = 4), in both the ipsi- and contralateral LC. Significantly larger numbers of double labelled neurones were counted in the LC of tooth-pulp-stimulated animals (representing around 30% of retrogradely labelled cells in LC) than in the LC of sham animals. They were found bilaterally, but with a clear, significant, ipsilateral (i.e. left) predominance. The present data offer an anatomical framework to understand how the LC is involved in the sensory processing of nociceptive information in the thalamus. For the first time, it is shown that nociceptive stimulation activates LC neurones projecting to the somatosensory thalamus. This suggests a new role for LC in modulating nociception within the thalamus.     

The volume-activated chloride current in endothelial cells from bovine pulmonary artery is not modulated by phosphorylation

We employed the patch-clamp technique to investigate the effects of various phosphorylation pathways on activation and modulation of volume-activated Cl- currents (I _Cl,vol) in cultured endothelial cells from bovine pulmonary arteries (CPAE cells). Half-maximal activation ofI _Cl,vol occurred at a hypotonicity of 27.5 ± 1.2%. Run-down of the current upon repetitive activation was less than 15% within 60 min. Stimulation of protein kinase C (PKC) by phorbol-12-myristate-13-acetate (PMA) or by (–)-indolactam did not affectI _Cl,vol. Down regulation of PKC activity by a 24-h preincubation of the cells with 0.2 mol/l PMA, or its inhibition by loading the cells with the specific inhibitory 19–31 pseudosubstrate peptide, did not influenceI _Cl,vol. Trifluoperazine and tamoxifen fully blockedI _cCl,vol with concentrations required for half-maximal inhibition of 3.0 and 2.4 mol/1 respectively. This inhibitory effect is probably not mediated by the calmodulin-antagonistic action of these compounds, because it occurs at free intracellular [Ca²⁺] of 50 nmol/l, which are below the threshold for calmodulin activation. The tyrosine kinase inhibitor herbimycin A (1 ol/1) and genistein (100 ol/1) did not affectI _Cl,vol Exposing CPAE cells to lysophosphatidic acid (1mol/1), an activator of p42 MAPkinase and the focal adhesion kinase p125^FAK in endothelial cells, neither evoked a Cl^– current nor affectedI _Cl,vol Neither wortmannin (10 mol/1), an inhibitor of MAP kinases and of PI-3 kinase, nor rapamycin (0.1 mmol/1), which interferes with the p70S6 kinase pathway, affectedI _Cl,vol Exposure of CPAE cells to heat or Na-arsenite, both activators of a recently discovered stress-activated tyrosine phosphorylation pathway, neither activated a current nor affected the hypotonic solution-induced Cl^– current. We conclude that none of the studied phosphorylation pathways is essential for the activation of the Cl^– current induced by hypotonicity.