Influence of Polishing Systems on Surface Roughness of Composite Resins: Polishability of Composite Resins.

OBJECTIVES: The objective of this in vitro study was to compare, with a threshold value of 200 nm, the surface roughness obtained when using 12 different polishing systems on four different composite resins (microfill, nanofill, and two nanohybrids). METHODS AND MATERIALS: A total of 384 convex specimens were made using Durafill VS, Filtek Supreme Ultra, Grandio SO, and Venus Pearl. After sandblasting and finishing with a medium-grit finishing disc, initial surface roughness was measured using a surface roughness tester. Specimens were polished using 12 different polishing systems: Astropol, HiLuster Plus, D♦Fine, Diacomp, ET Illustra, Sof-Lex Wheels, Sof-Lex XT discs, Super-Snap, Enhance/Pogo, Optrapol, OneGloss and ComposiPro Brush (n=8). The final surface roughness was measured, and data were analyzed using two-way analysis of variance. Pairwise comparisons were made using protected Fisher least significant difference. RESULTS: There were statistical differences in the final surface roughness between polishing systems and between composite resins (p<0.05). The highest surface roughness was observed for all composite resins polished with OneGloss and ComposiPro Brush. Enhance/Pogo and Sof-Lex Wheels produced a mean surface roughness greater than the 200-nm threshold on Filtek Supreme Ultra, Grandio SO, and Venus Pearl. Data showed that there was an interaction between the composite resins and the polishing systems. CONCLUSIONS: A single polishing system does not perform equally with all composite resins. Except for Optrapol, multi-step polishing systems performed generally better than one-step systems. Excluding Enhance/Pogo, diamond-impregnated polishers led to lower surface roughness. Durafill VS, a microfill composite resin, may be polished more predictably with different polishers.

[Miller Fisher syndrome, internal and external ophthalmoplegia after flu vaccination]. / Síndrome de Miller Fisher, oftalmoplejía interna y externa tras vacunación antigripal.

CASE REPORT: Miller Fisher Syndrome (MFS) is the most frequent variant of the Guillain-Barré Syndrome. It is characterised by the classic triad of ophthalmoplegia, ataxia and areflexia. We present a case of a patient who developed these clinical findings 5 days after flu vaccination. DISCUSSION: Miller Fisher Syndrome is an unusual condition seen in ophthalmologic clinical practice. Although respiratory and digestive infections have been reported as antecedent infectious agents in MFS, it has not previously been described in relation to the flu vaccine.

Honey protects against wings posture error and molecular changes related to mitochondrial pathways induced by hypoxia/reoxygenation in adult Drosophila melanogaster.

We conducted an investigation to evaluate the effects of Brazilian Pampa biome honey and its major phenolic compounds on the development of an erected wings posture phenotype and related mitochondrial aspects induced by Hypoxia/Reoxygenation (H/R) in Drosophila melanogaster. Flies were pre-treated for 3 days with a 10% honey solution and different concentrations of caffeic acid and ρ-coumaric acid and then submitted to hypoxia for 3 h. We observed that after reoxygenation, some flies acquired an erected wings posture and that this feature may be related to mortality. In addition, H/R induced down-regulation of ewg mRNA expression, which could be associated to the observed complex phenotype. H/R also caused a dysregulation in opa1-like, ldh and diap genes expression and reduced O2 fluxes in flie's mitochondria. Honey mitigated opa1-like mRNA expression changes provoked by H/R. Differently from honey, caffeic and ρ-coumaric acids displayed no protective effects. In conclusion, we report for the first time the protective effects of honey against complex phenotypes and mitochondrial changes induced by H/R in adult flies.

Non-pathogenic Entamoeba histolytica: functional and biochemical characterization of a monoxenic strain.

We have cultured under monoxenic conditions and characterized an Entamoeba histolytica clone, MAV-I CINVESTAV (MAV-I), obtained from feces from an asymptomatic carrier. The clone shows the non-pathogenic E. histolytica zymodeme type I, which did not change through the process of monoxenization. Clone MAV-I was non-pathogenic in both in vivo and in vitro tests, and it did not have a functional 112-kDa adhesin. As far as we know, this is the first non-pathogenic monoxenic strain reported. Clone A (strain HM1:IMSS), a highly virulent clone with pathogenic zymodeme type II, and which has the 112-kDa adhesin, was used as a control. Protein patterns from both clones were almost identical in one-dimensional gels. In two-dimensional gels, differences in high-molecular-weight proteins were detected. Clone MAV-I adhered and phagocytosed only 12% of the red blood cells adhered and phagocytosed by clone A. MAV-I trophozoites did not destroy cell culture monolayers and did not produce hepatic abscesses in hamsters. They also showed deficiency in protease activity. The absence of virulence in clone MAV-I correlated directly with the absence of a functional 112-kDa adhesion, supporting the role that this protein plays in virulence.

Thyrotropin-releasing hormone-induced down-regulation of pyroglutamyl aminopeptidase II activity involves L-type calcium channels and cam kinase activities in cultures of adenohypophyseal cells.

Released thyrotropin-releasing hormone (TRH) is inactivated by a narrow specificity ectopeptidase, pyroglutamyl aminopeptidase II (PPII), present in brain and lactotrophs. Various hypothalamic/paracrine factors, including TRH, slowly (in hours) regulate the activity of PPII on the surface of adenohypophyseal cells. TRH-induced down-regulation was mimicked by protein kinase C (PKC) activation but was not affected by inhibition of PKC. Adenylate cyclase activation can also down-regulate PPII. The purpose of this study was to identify elements of the transduction pathway used by TRH to regulate PPII activity. In primary cultures of female adenohypophyseal cells, activation of the stimulatory G protein or adenylate cyclase produced an effect additive to that of TRH; inhibition of protein kinase A activity did not interfere with TRH action. However, regulation of PPII activity by TRH was inhibited by a phospholipase C beta inhibitor or chelation of intracellular calcium. L-type calcium channels (LCC) agonists mimicked TRH action and their effect was not additive with that of TRH. Antagonists of LCC channels and inhibitors of calmodulin or calcium/calmodulin-dependent protein kinase blocked TRH action. Therefore, TRH-induced calcium entry through L-type calcium channels and the activity of calcium/calmodulin-dependent protein kinase are required for TRH effect on PPII activity in primary cultures of adenohypophyseal cells. This pathway may coregulate PPII and prolactin biosynthesis in response to TRH.

Multiple hypothalamic factors regulate pyroglutamyl peptidase II in cultures of adenohypophyseal cells: role of the cAMP pathway.

In the adenohypophysis, thyrotrophin-releasing hormone (TRH) is inactivated by pyroglutamyl peptidase II (PPII), a TRH-specific ectoenzyme localized in lactotrophs. TRH slowly downregulates surface PPII activity in adenohypophyseal cell cultures. Protein kinase C (PKC) activation mimics this effect. We tested the hypothesis that other hypothalamic factors controlling prolactin secretion could also regulate PPII activity in adenohypophyseal cell cultures. Incubation for 16 h with pituitary adenylate cyclase activator peptide 38 (PACAP; 10(-6) M) decreased PPII activity. Bromocryptine (10(-8) M), a D2 dopamine receptor agonist, or somatostatin (10(-6) M) stimulated enzyme activity and blocked the inhibitory effect of [3-Me-His2]-TRH, a TRH receptor agonist. Bromocryptine and somatostatin actions were suppressed by preincubation with pertussis toxin (400 ng ml(-1)). Because these hypophysiotropic factors transduce some of their effects using the cAMP pathway, we analysed its role on PPII regulation. Cholera toxin (400 ng ml(-1)) inhibited PPII activity. Forskolin (10(-6) M) caused a time-dependent decrease in PPII activity, with maximal inhibition at 12-16 h treatment; ED50 was 10(-7) M. 3-isobutyl-1-methylxanthine or dibutiryl cAMP, caused a dose-dependent inhibition of PPII activity. These data suggest that increased cAMP down-regulates PPII activity. The effect of PACAP was blocked by preincubation with H89 (10(-6) M), a protein kinase A inhibitor, suggesting that the cAMP pathway mediates some of the effects of PACAP. Maximal effects of forskolin and 12-O-tetradecanoylphorbol 13-acetate were additive. PPII activity, therefore, is independently regulated by the cAMP and PKC pathways. Because most treatments inhibited PPII mRNA levels similarly to PPII activity, an important level of control of PPII activity by these factors may be at the mRNA level. We suggest that PPII is subject to 'homologous' and 'heterologous' regulation by elements of the multifactorial system that controls prolactin secretion.

The narrow specificity pyroglutamate amino peptidase degrading TRH in rat brain is an ectoenzyme.

In order to determine the pathway of extracellular metabolism of the thyrotropin releasing hormone (pyroglu-his-proNH(2)) in brain, the topographical organization of pyroglutamate aminopeptidase II on the plasma membrane was investigated. Its activity was only slightly increased when intact brain synaptosomes were lysed by osmotic shock or detergent treatment. Trypsin treatment of intact synaptosomes destroyed 70-80% of enzyme activity without affecting lactate dehydrogenase. Pyroglutamate aminopeptidase II activity was present in primary cultures of foetal mice cortical cells. It was detected in intact cells, was not released by the cells and its activity was not increased by saponin pretreatment. Trypsin treatment of the cells reduced pyroglutamate aminopeptidase II by 70% but did not affect pyroglutamate aminopeptidase I and lactate dehydrogenase. These data support that brain pyroglutamate aminopeptidase II is an ectoenzyme. They suggest that this enzyme could be responsible for thyrotropin releasing hormone extracellular catabolism in brain.

Subcellular distribution of the enzymes degrading thyrotropin releasing hormone and metabolites in rat brain.

In order to further understand the role of enzymes degrading Thyrotropin Releasing Hormone (TRH, pglu-his-proNH(2)) and metabolites, we studied their subcellular distribution in rat brain. Brain tissue was homogenized in 0.32 M sucrose, tris-HCl 0.01 M pH 7.4 and fractionated by differential and discontinuous gradient centrifugation; [(3)H]pro-TRH was incubated with the various subcellular fractions and the extent of degradation of each metabolite was measured after separation by thin layer chromatography. Several markers were simultaneously measured (lactate dehydrogenase, 5?-nucleotidase and hexosaminidase) to determine the pattern of distribution of the subcellular organelles. The post-proline cleaving enzyme responsible for pglu-his-pro formation and pyroglutamate amino-peptidase (which requires sulphydryl compounds for maximal activity) were found in cytosol but were barely detectable in the soluble component of synaptosomes; pyroglutamate aminopeptidase (dependent on metals) and post-proline dipeptidyl amino peptidase were found on the membranes of synaptosomes; imido peptidase was not enriched in any particular fraction. These data are consistent with the hypothesis that membrane-bound pyroglutamate aminopeptidase is responsible for TRH degradation once released into the synaptic cleft and that the post-proline dipeptidylaminopeptidase may participate in the extracellular catabolism of his-proNH(2) before it cyclizes to his-pro-DKP. They also suggest that post-proline cleaving enzyme and soluble pyroglutamate aminopeptidase may not play an important role in the regulation of TRH levels in nerve endings.

Regional distribution of in vitro release of thyrotropin releasing hormone in rat brain.

To increase our knowledge of the TRH functions in brain and the processes of TRH compartmentalization and release, we studied the in vitro release of endogenous TRH in different brain areas. We also determined the correlation between TRH levels and release under both basal and stimulated conditions. TRH concentration was measured in tissues and media by specific radioimmunoassay. TRH-like material detected in olfactory bulb and hypothalamic incubates (basal or K+ stimulated) were shown to be chromatographically identical to synthetic TRH. Different brain regions showed high variability in the basal release of TRH (1-20% of tissue content). This suggests the existence of different pools. The response to depolarizing stimulus (56 mM K+) was significant only in the following regions: median eminence, total hypothalamus, preoptic area, nucleus accumbens-lateral septum, amygdala, mesencephalon, medulla oblongata and the cervical region of the spinal cord. These regions have been shown to contain a high number of receptors, a high concentration of TRH nerve endings and are susceptible to TRH effects. These results support the hypothesis that TRH functions as neuromodulator in these areas.

Regional distribution of pyroglutamyl peptidase II in rabbit brain, spinal cord, and organs.

Pyroglutamyl peptidase II (PPII) is a narrow specificity ectoenzyme that degrades thyrotropin-releasing hormone (TRH). We detected the enzyme in the brain of various mammals, with highest specific activity in rabbit brain. In this species, activity was heterogeneously distributed in the central nervous system. There was a 28-fold difference between regions of highest and lowest PPII activity. Enzyme activity was highest in the olfactory bulb and posterior cortex. In the spinal cord, activity was low but unevenly distributed, with highest values detected in the thoracic (T) region. Segments T1 and T2 activities were particularly high. Other organs contained low or undetectable levels of activity. The levels of TRH-like immunoreactivity (TRH-LI) in spinal cord segments were greatest in T3-T4 and lumbar L2-L6. Low concentrations were found in T1 and T9-T12. There was a partial correlation between the distribution of PPII activity and TRH receptors but not with TRH-LI levels. These results demonstrate that PPII is predominantly a central nervous system enzyme, and they support the hypothesis that PPII is responsible for degrading TRH released into the synaptic cleft.

The effects of alprazolam on corticotropin-releasing factor neurons in the rat brain: implications for a role for CRF in the pathogenesis of anxiety disorders.

Considerable evidence indicates that corticotropin-releasing factor (CRF) is responsible for integrating not only the endocrine, but the autonomic and behavioral responses of an organism to stress. We have investigated the effects of the anxiolytic triazolobenzodiazepine, alprazolam, on the activity of the hypothalamic-pituitary-adrenal (HPA) axis and of CRF neurons following acute and chronic administration. In addition, because many of the signs and symptoms observed in animals and humans following abrupt discontinuation of benzodiazepines resemble those of the stress response, we examined the effect of alprazolam withdrawal on CRF neurons and HPA axis activity. Alprazolam decreases CRF concentrations in the locus coeruleus 0.5-3.0 hours following acute injection. Similarly, chronic (14 days) alprazolam administration also results in decreased CRF concentrations in the locus coeruleus. CRF concentrations return to control values 24 hours following abrupt alprazolam withdrawal. Moreover, abrupt alprazolam withdrawal results in increased plasma ACTH and corticosterone concentrations and decreased anterior pituitary CRF receptor concentrations 24 hours following drug discontinuation. Thus, abrupt alprazolam withdrawal profoundly activates the HPA axis. These indices of HPA axis activity return to control values by 48 hours post-withdrawal. These actions of alprazolam on CRF neurons are opposite to those observed following acute or chronic stress. These results support the hypothesis that CRF-containing neurons innervating the locus coeruleus may be involved in the pathogenesis of anxiety, and in the actions of clinically efficacious anxiolytics.

Pyroglutamyl peptidase II inhibition specifically increases recovery of TRH released from rat brain slices.

Pyroglutamyl peptidase II (EC 3.4.19-) is a highly specific membrane-bound thyrotropin releasing hormone (TRH) degrading enzyme. To study the functional significance of pyroglutamyl peptidase II in TRH degradation, we synthesized the reversible inhibitor N-1-carboxy-2-phenylethyl (Nimbenzyl)-histidyl-beta-naphthylamide (CPHNA). CPHNA inhibited the enzyme with a Ki of 8 microM, but had no effect no TRH receptors or no prolyl endopeptidase (EC 3.4.21.26). It weakly inhibited cytosolic pyroglutamyl peptidase I (EC 3.4.19.3). CPHNA at a concentration of 10(-4) M increased both the basal and potassium stimulated recovery of TRH released from hypothalamic slices by approximately two-fold. An even higher recovery was observed in slices from brain regions with relatively high levels of pyroglutamyl peptidase II. CPHNA had no effect on the basal recovery of gamma-aminobutyric acid or Met-enkephalin released from brain slices but decreased the potassium stimulated recovery of both Metenkephalin and gamma-aminobutyric acid. These data further support the involvement of pyroglutamyl peptidase II in the extracellular inactivation of brain TRH.

Ontogenesis of pyroglutamyl peptidase II activity in rat brain, adenohypophysis and pancreas.

Pyroglutamyl peptidase II (PPII; E.C. 3.4.19.-) is a highly specific membrane-bound ectoenzyme degrading thyrotropin releasing hormone (TRH). The ontogenesis of this enzyme was measured in rat brain regions, adenohypophysis and pancreas. In hypothalamus PPII activity was maximal at day 8 postnatal, decreasing to adult values at day 45. The postnatal ontogenic patterns in posterior cerebral cortex and hypothalamus were similar. In olfactory bulb, two peaks of activity were observed (3th and 22nd day) while in adenohypophysis it appeared only at day 8, increased to day 30, decreasing thereafter to adult values.

Pyroglutamyl peptidase II activity is not in the processes of bulbospinal TRHergic neurons.

Pyroglutamyl peptidase II (PPII) is a neuronal ectoenzyme involved in released thyrotropin-releasing hormone (TRH) inactivation. In an attempt to define if it is present in the pre or postsynaptic membrane, we induced neuronal degeneration of serotonin-TRHergic cells that project from raphe nuclei to the spinal cord. 2-4 weeks after intracisternal injection of 5,7-dihydroxytryptamine, TRH levels decreased over 70% in the cervical, thoracic or lumbar regions of spinal cord. In contrast, no change of PPII activity was observed. Longer times after injection (6-8 weeks), a 59-66% increase in activity was detected in the lumbar region. These data suggest that PPII is not localized in these TRHergic neurons but probably in the target cells.

Carotenoid content of chlorophycean microalgae: factors determining lutein accumulation in Muriellopsis sp. (Chlorophyta).

Fifteen strains of chlorophycean microalgae have been investigated with regard to their carotenoid profile. Lutein, beta-carotene and violaxanthin were present in virtually all of the strains, lutein, in general, being the most abundant carotenoid, whereas canthaxanthin and astaxanthin were found in some strains only. Chlorella fusca SAG 211-8b, Chlorococcum citriforme SAG 62.80, Muriellopsis sp., Neospongiococcum gelatinosum SAG B 64.80 and Chlorella zofingiensis CCAP 211/14 exhibited high lutein levels, the latter strain containing in addition substantial amounts of astaxanthin. Muriellopsis sp. was further characterized, since besides a high lutein content (up to 35 mg l(-1) culture), it had the highest growth rate (up to 0.17-0.23 h(-1)) and maximal standing cell density (up to 8 x 10(10) cells l(-1) culture). These levels of lutein are in the range of those reported for astaxanthin in Haematococcus and for beta-carotene in Dunaliella, microalgae of recognized interest for the production of these carotenoids. Lutein content of Muriellopsis sp. increased during the exponential phase of growth, with the highest value being recorded in the early stationary phase. Maximum levels of lutein in Muriellopsis sp. cultures were recorded at 20-40 mM NaNO3, 2-100 mM NaCl, 460 micromol photon m(-2) s(-1), pH 6.5 and 28 degrees C, conditions which were, in general, also optimal for cell growth. Growth-limiting conditions, such as pH values of 6 or 9 and a temperature of 33 degrees C, were found to stimulate carotenogenesis in Muriellopsis sp. This strain represents a potential source of lutein, a commercially interesting carotenoid of application in aquaculture and poultry farming, as well as in the prevention of cancer and diseases related to retinal degeneration.

Lutein production by Muriellopsis sp. in an outdoor tubular photobioreactor.

The effect of dilution rate, mixing and daily solar cycles on lutein and biomass productivity of the green unicellular alga Muriellopsis sp. has been studied, throughout the year, in an outdoor tubular photobioreactor. Highest productivity values, for both lutein (about 180 mg m(-2) per day) and biomass (about 40 g (dry weight) m(-2) per day) were achieved on May and July. Values for the optimal dilution rate varied, being lower in May (0.06 h(-1)) than in November (0.09 h(-1)). Similar values for photosynthetic efficiency (about 4%) were recorded throughout the year, indicating that optimization of culture conditions was achieved for each experimental period. Along the daily solar cycle, there was a fast increase of lutein content of Muriellopsis sp. in response to irradiance during the early hours of daytime, with maximal lutein content (about 6 mg (g dry weight)(-1)) being recorded at noon, and decreasing slowly, thereafter. An increase in cell growth was observed following the establishment of maximum lutein/chlorophyll ratio, which might indicate a role for lutein in protecting cells from photodamage.

Post-traumatic stress disorder.

In the past few years, there has been a growing interest in the neurobiology of PTSD. A number of models have been proposed including possible structural changes. We have now seen the appearance in the literature of controlled and double-blind trials. Of interest are positive outcome studies associated with the use of drugs with serotonergic actions for a period of 8 weeks. Symptom relief provided by pharmacotherapy enables the patient to participate more thoroughly in individual, behavioral, or group therapy.

Viscoelastic-injecting cystotome.

This continuous curvilinear capsulorhexis (CCC) technique is for use in complicated surgical cases such as when the anterior chamber is shallow, the red reflex is not good, or eye movements are present. This technique is easier and safer in such cases because it uses a cystotome connected to a viscoelastic syringe. First, the anterior chamber is filled with viscoelastic material using a conventional cannula. The cannula is replaced with a bent needle (or cystotome), and the CCC is performed in the usual way. This instrument allows the surgeon to inject small amounts of viscoelastic material exactly where and when it is needed. The anterior chamber remains deep while the CCC is performed, and the anterior capsule tear is done in a more controlled fashion.

Effect of in vitro maedi-visna virus infection on adherence and phagocytosis of staphylococci by ovine cells.

This work was aimed at studying the effect of maedi-visna virus (MVV) infection in vitro on the ability of sheep cells to adhere to staphylococci (Staphylococcus aureus and Staphylococcus epidermidis), and phagocytose these bacteria. Adherence was studied in sheep choroid plexus cells (SCPC) using an ELISA test and phagocytosis was studied in pulmonary alveolar macrophages (PAM) by chemiluminescence. A 5- and 7-day of in vitro MVV infection resulted in syncytium formation and a significant increased adherence (P < 0.01) of SCPC to bacteria. SCPC endogenous fibronectin was significantly higher (P < 0.01) on days 5 and 7 than on day 0 of MVV infection. A significantly decreased phagocytosis (P < 0.05) was also observed on days 5 and 7 of MVV infection in PAM when compared to MVV-free controls. Comparatively, phagocytosis was highest for S. aureus non-slime producing strains, followed by S. epidermidis, and S. aureus slime producing strains, in that order. Finally, increased expression of both, class I and class II major histocompatibility antigens was also observed in MVV-infected PAM on days 5 and 7, whereas SCPC only demonstrated upregulation of MHC class I. These results, indicative of an alteration of some cell functions in MVV-infected cells, may help to understand interactions between MVV-infected cells and bacteria in simultaneous infections and may provide clues to the possible in vivo interactions of both pathogens.

Early detection of maedi-visna (ovine progressive pneumonia) virus seroconversion in field sheep samples.

The aim of this work was to investigate whether an enzyme-linked immunosorbent assay (ELISA) was useful for early detection of maedi-visna virus (MVV) infection in sheep under field conditions. An ELISA based on p25 recombinant protein and a gp46 synthetic peptide was used. Sequentially obtained serum samples (n = 1,941) were studied for 4 years. ELISA results were compared with those of the agar gel immunodiffusion (AGID) test, and results of both tests were compared with a reference result established using consensus scores for at least 2 of 3 serologic techniques (AGID, ELISA, and western blotting, which was used to resolve result discrepancies between the other 2 techniques). A total of 247 discrepancies were observed between ELISA and AGID. Of these, 131 were due to an earlier detection of 120 sera by the ELISA and 11 sera by AGID. The remaining discrepancies (116) were due to the presence of false reactions in both tests. Fewer false-negative results were found by ELISA than with AGID (6 vs. 69 sera, respectively), whereas the number of false-positive results was virtually the same for ELISA and AGID (21 vs. 20, respectively). In relation to the reference result, ELISA sensitivity and specificity were 97.8% and 98.2%, respectively, whereas values for AGID were 76.3% and 98.3%, respectively. The agreement between ELISA and the reference result was higher than that between AGID and the reference result (K value: 0.96 and 0.77, respectively). A variation in the ELISA signal (based on optical density) was observed during the study period, suggesting different antibody levels throughout the animal's life. The ELISA was useful for detecting MVV-infected sheep in field conditions and has potential for use in control and eradication programs.