Effects of Ethanol on Synthesis of Prostaglandin F2 alpha in Bovine Females

Ethanol stimulates the production of prostaglandins in many species. The purpose of this study was to verify the effect of ethanol on the production of prostaglandin F2 alpha (PGF2 alpha) and luteolysis in bovine females. In the first experiment, Holstein cows at day 17 of the oestrous cycle were treated with 100% ethanol (0.05 ml/kg of body weight, IV; n = 5), saline (0.05 ml/kg of body weight, IV; n = 4) or synthetic prostaglandin (150 mu g of D-cloprostenol/cow, IM; n = 4). The plasma concentrations of 13, 14-dihydro-15-keto PGF2 alpha (PGFM; the main metabolite of PGF2 alpha measured in the peripheral blood) were assessed by radioimmunoassay (RIA). There was an acute release of PGFM in response to ethanol comparing to other treatments (p <= 0.05). However, only cows treated with PGF2 alpha underwent luteolysis. In the second experiment, endometrial explants of cross-bred beef cows (n = 4) slaughtered at day 17 of the oestrous cycle were cultured for 4 h. During the last 3 h, the explants were cultured with medium supplemented with 0, 0.1, I, 10 or 100 mu l of 100% ethanol/ml. Medium samples were collected at hours 1 and 4 and concentrations of PGF2 alpha were measured by RIA. Ethanol did not induce PGF2 alpha production by the endometrium. In conclusion, ethanol does not cause luteolysis in cows because it stimulates production of PGF2 alpha in extra-endometrial tissues.

The influence of erbium:yttrium-aluminum-garnet laser ablation with variable pulse width on morphology and microleakage of composite restorations

The objective of this study was to evaluate the influence of various pulse widths with different energy parameters of erbium:yttrium-aluminum-garnet (Er:YAG) laser (2.94 mu m) on the morphology and microleakage of cavities restored with composite resin. Identically sized class V cavities were prepared on the buccal surfaces of 54 bovine teeth by high-speed drill (n = 6, control, group 1) and prepared by Er:YAG laser (Fidelis 320A, Fotona, Slovenia) with irradiation parameters of 350 mJ/ 4 Hz or 400 mJ/2 Hz and pulse width: group 2, very short pulse (VSP); group 3, short pulse (SP); group 4, long pulse (LP); group 5, very long pulse (VLP). All cavities were filled with composite resin (Z-250-3 M), stored at 37A degrees C in distilled water, polished after 24 h, and thermally stressed (700 cycles/5-55A degrees C). The teeth were impermeabilized, immersed in 50% silver nitrate solution for 8 h, sectioned longitudinally, and exposed to Photoflood light for 10 min to reveal the stain. The leakage was evaluated under stereomicroscope by three different examiners, in a double-blind fashion, and scored (0-3). The results were analyzed by Kruskal-Wallis test (P > 0.05) and showed that there was no significant differences between the groups tested. Under scanning electron microscopy (SEM) the morphology of the cavities prepared by laser showed irregular enamel margins and dentin internal walls, and a more conservative pattern than that of conventional cavities. The different power settings and pulse widths of Er:YAG laser in cavity preparation had no influence on microleakage of composite resin restorations.

Polysaccharide fraction of Agaricus brasiliensis avoids tumor-induced IL-10 production and changes the microenvironment of subcutaneous Ehrlich adenocarcinoma

Subcutaneous Ehrlich tumor-bearing mice were treated with in situ inoculation of a P-glucan-rich extract of Agaricus brasiliensis (ATF), which reduced tumor growth. Histopathological analysis showed that the tumor masses of control mice (Ehr) presented giant tumor cells and many mitotic figures whereas the tumor tissue obtained from ATF-treated animals (Ehr-ATF) presented a lower frequency of both mitotic and giant cells, associated with a higher frequency of apoptotic cells than Ehr. Analysis of the lymphoproliferative activity of spleen cells showed that the treatment had a suppressive rather than a stimulatory effect. Spleen cells of the Ehr group produced higher in vitro levels of IL-10 than normal controls and this occurrence was partially avoided by treatment with ATF. Analysis of cytokine production by tumor-infiltrating cells (ELISpot) showed that ATF induced a higher number of IFN-gamma-producing cells at 7 and 14 days as well as reduction of IL-10-secreting cells at the latter time. Confocal microscopy analysis showed higher intensity of labeling of CD4+ and Mac-3+ cells in ATF-treated mice. Analysis of in situ expression of angiogenic growth factors showed a slight decrease of FGF-2 mRNA in Ehr-ATF animals (7th day) but not of VEGF-A or TGF-beta expression. This fraction could not directly lyse either lymphocytes or tumor cells and we speculate that antitumor effect of ATF could be due to induction of a selective migration of immunocompetent cells from the spleen to the tumor site and to the switch of cytokine production. (C) 2009 Elsevier Inc. All rights reserved.

Functional polymorphisms in the promoter of the matrix metalloproteinase-9 (MMP-9) gene are not linked with significant plasma MMP-9 variations in healthy subjects

Background: Matrix metalloproteinase-9 (MMP-9) is involved in the degradation of the extracellular matrix during physiological and pathological processes. Two functional polymorphisms [C(-1562)T and microsatellite (CA)(13-25)] in the promoter region of the MMP-9 gene have been associated with several diseases. The aim of this study was to examine whether these MMP-9 polymorphisms and haplotypes are linked with plasma MMP-9 variations in healthy subjects. Methods: We studied 177 healthy male white volunteers (age range 20-55 years) who were non-smokers and not taking any medication. Genomic DNA was extracted from whole blood and genotypes for the C(-1562)T and the microsatellite (CA)(n) polymorphisms were determined. MMP-9 levels were measured in plasma samples by gelatin zymography. Results: The frequency of the alleles C and T for the C(-1562)T polymorphism were 90% and 10%, respectively. The frequency of the alleles with less than 21 CA repeats Q and with 21 repeats or higher (H) were 47% and 53%, respectively. We found no differences in plasma MMP-9 levels among the genotype groups or among different haplotypes (all p > 0.05). Conclusions: These findings suggest that functional polymorphisms in the promoter of the MMP-9 gene are not linked with significant plasma MMP-9 variations in healthy subjects.

Inverse relationship between markers of nitric oxide formation and plasma matrix metalloproteinase-9 levels in healthy volunteers

Background: Nitric oxide (NO) is a major regulator of cardiovascular homeostasis and has anti-atherogenic properties. Reduced NO formation is associated with endothelial dysfunction and with cardiovascular risk factors. Although NO downregulates the expression and activity of the pro-atherogenic enzyme matrix metalloproteinase-9 (MMP-9), no previous clinical study has examined whether endogenous NO formation is inversely associated with the circulating levels of pro-MMP-9, which are associated with cardiovascular events. We examined this hypothesis in 175 healthy male subjects who were non-smokers. Methods: To assess NO bioavailability, the plasma concentrations of nitrite, nitrate, and cGMP were determined using an ozone-based chemiluminescence assay and an enzyme immunoassay. Pro-MMP-9 and pro-MMP-2 levels were measured in plasma samples by gelatin zymography. Results: We found significant negative correlations between pro-MMP-9 levels and plasma nitrite (P=0.035, rs=-0.159), nitrate (P=0.040, rs=-0.158), and cGMP (P=0.011, rs=-0.189) concentrations. However, no significant correlations were found between pro-MMP-2 levels and the plasma concentrations of markers of NO bioavailability (all P>0.05). Conclusions: There is an inverse relationship between markers of NO formation and plasma MMP-9 levels. This finding may shed some light on the possible mechanisms involved in the increased cardiovascular risk of apparently healthy subjects with low NO bioavailability or high circulating levels of pro-MMP-9. (C) 2008 Elsevier B.V. All rights reserved.

Ultrastructural Analysis of Radicular Dentine Surface Submitted to CO(2) Laser at Different Parameters

Background: There are no reported studies comparing different parameter settings of the CO(2) laser and irradiation direction considering their effect on the morphology of radicular dentine surface. Purpose: To evaluate the alterations of radicular dentine (cervical, middle, and apical thirds) irradiated with CO(2) laser at different potencies and irradiation directions. Study Design: Roots of 35 canines were prepared and randomly distributed according to the laser potency: GI: no laser treatment (control) (n = 5); GII, 2 W (n = 10); GIII: 4 W (n = 10); GIV: 6 W (n = 10). Each group (excepting GI) was divided in two subgroups according to the irradiation distance (n = 5): (A) parallel and (B) perpendicular to the root canal walls. The roots were splited longitudinally and analyzed by scanning electron microscopy in a qualiquatitative way. The scores were submitted to Kruskal-Wallis and Dunn`s tests. Results: No significant statistical differences were observed among root canal thirds (P > 0.05). The specimens irradiated with 2 W were statistically different (P < 0.05) from those irradiated with 4 and 6 W, which were statistically similar between themselves (P > 0.05). With 2, 4, and 6 W at in parallel irradiation and 2 W in perpendicular direction, the surface showed a fissured aspect. With 4 W in perpendicular direction and 6 W in parallel and perpendicular direction, surface was modified by laser action and exhibited fused areas. Conclusions: The intensity of the effects is dependent on the laser-irradiation dosimetries. Alterations were more intense when higher parameters were used. Microsc. Res. Tech. 72:737-743, 2009. (C) 2009 Wiley-Liss, Inc.

Morphological Alterations of Radicular Dentine Pretreated With Different Irrigating Solutions and Irradiated With 980-nm Diode Laser

Background: The topographical features of intraradicular dentine pretreated with sodium hypochlorite (NaOCl) or ethylenediamine tetraacetic acid (EDTA) followed by diode laser irradiation have not yet been determined. Purpose: To evaluate the alterations of dentine irradiated with 980-nm diode laser at different parameters after the surface treatment with NaOCl and EDTA. Study design: Roots of 60 canines were biomechanically prepared and irrigated with NaOCl or EDTA. Groups were divided according to the laser parameters: 1.5 W/CW; 1.5 W/100 Hz; 3.0 W/CW; 3.0 W/100 Hz and no irradiation (control). The roots were splited longitudinally and analyzed by scanning electron microscopy (SEM) in a quali-quatitative way. The scores were submitted to two-way Kruskal-Wallis and Dunn`s tests. Results: The statistical analysis demonstrated that the specimens treated only with NaOCl or EDTA (control groups) were statistically different (P < 0.05) from the laser-irradiated specimens, regardless of the parameter setting. The specimens treated with NaOCl showed a laser-modified surface with smear layer, fissures, and no visible tubules. Those treated with EDTA and irradiated by laser presented absence of smear layer, tubules partially exposed and melting areas. Conclusions: The tested parameters of 980-nm diode laser promoted similar alterations on dentine morphology, dependent to the type of surface pretreatment. Microsc. Res. Tech. 72:22-27, 2009. (C) 2008 Wiley-Liss, Inc.

The identification of the transcriptional regulator CRP in Aeromonas hydrophila JMP636 and its involvement in amylase production and the 'acidic toxicity' effect

Aims: The physiological examination of amylase production by Aeromonas hydrophila JMP636 and identification of the mechanism of regulation. Methods and Results: Aeromonas hydrophila JMP636 was grown with single, then dual carbon sources; the growth cycle was followed and amylase activity throughout was monitored. The levels of cAMP, a known secondary messenger for the regulatory gene crp, were also examined. Amylase activity was regulated by catabolite repression. Physiological studies revealed that JMP636 exhibited both diauxic growth, with two carbon sources, and the 'acid toxicity' effect on glucose. The crp gene was cloned, expressed and inactivated from the JMP636 chromosome. Catabolite repression of amylase production and the 'acid toxicity' effect both require crp and were linked to cAMP levels. Conclusions: Regulation of amylase production was predicted to follow the model CRP-mediated cAMP-dependent Escherichia coli catabolite regulation system. Significance and Impact of the Study: This work provides an understanding of the physiology of the opportunistic pathogen Aer. hydrophila through identification of the mechanism of catabolite repression of amylase production and the existence of crp within this cell. It also provides a broader knowledge of global gene regulation and suggests regulatory mechanisms of other Aer. hydrophila gene/s.

Exposure to automotive pollution increases plasma susceptibility to oxidation

Low-density lipoprotein oxidation is implicated in the development of atherosclerosis. Plasma susceptibility to oxidation may be used as a marker of low-density lipoprotein oxidation and thus predict atherosclerotic risk. In this study the authors investigated the relationship between plasma susceptibility to oxidation and exposure to automotive pollution in a group of automobile mechanics (n = 16) exposed to high levels of automotive pollution, vs. matched controls (n = 13). The authors induced plasma oxidation by a free radical initiator and they determined susceptibility to oxidation by (1) change in absorbance at 234 nm, (2) lag time to conjugated diene formation, and (3) linear slope of the oxidation curve. Mechanics had significantly higher values (mean standard error) for change in absorbance (1.60 +/- 0.05 vs. 1.36 +/- 0.05; p < .002), and slope (1.6 x 10(-3) +/- 0.1 x 10(-3) vs. 1.3 x 10(-3) +/- 0.1 x 10(-3); p < .001), compared with controls. These results indicate that regular exposure to automotive pollutants increases plasma susceptibility to oxidation and may, in the long term, increase the risk of developing atherosclerosis.

Clarithromycin and prednisolone inhibit cytokine production in chronic rhinosinusitis

Objectives. Long-term, low-dose macrolide therapy is effective in the treatment of chronic rhinosinusitis. The mechanism of its anti-inflammatory effect and how this differs from corticosteroids remains unclear. The effect of clarithromycin and prednisolone on interleukin-5, interleukin-8, and granulocyte-macrophage colony-stimulating factor production by cultured chronic sinusitis nasal mucosa was examined in the study. Study Design and Methods. Nasal mucosa was obtained from 11 patients with chronic sinusitis. This tissue was cultured for 24 hours in the presence of clarithromycin or prednisolone at a variety of concentrations. Cytokine levels were determined by enzyme-linked immunoassay. Results. Clarithromycin and prednisolone each produced significant reductions in interleukin-5, interleukin-8, and granulocyte-macrophage colony-stimulating factor production. There was no significant difference between the effects of clarithromycin and prednisolone. Conclusion: Macrolide antibiotics are capable of inhibiting pro-inflammatory cytokine production in vitro and are as potent as prednisolone. This mechanism is likely to be at least partly responsible for the clinical efficacy of macrolide antibiotics in chronic rhinosinusitis. Key Words. Macrolide, prednisolone, sinusitis, enzyme-linked immunosorbent assay, cytokine.

Plasma growth hormone and growth hormone-binding protein during development in the marsupial brushtail possum (Trichosurus vulpecula)

Plasma concentrations of growth hormone (GH) were measured in the brushtail possum (Trichosurus vulpecula) pouch young from 25 through to 198 days post-partum (n=71). GH concentrations were highest early in pouch life (around 100 ng/ml), and thereafter declined in an exponential fashion to reach adult concentrations (10.8 +/- 1.8 ng/ml; n=21) by approximately 121-145 days post-partum, one to two months before the young is weaned. Growth hormone-binding protein (GHBP), which has been shown to modify the cellular actions of GH in eutherian mammals, was identified for the first time in a marsupial. Based on size exclusion gel filtration, possum GHBP had an estimated molecular mass of approximate to 65 kDa, similar to that identified in other mammalian species, and binding of I-125-labelled human GH (hGH) was displaced by excess hGH (20 mug). An immunoprecipitation method, in which plasma GHBP was rendered polyethylene glycol precipitable with a monoclonal antibody to the rabbit GHBP/GH receptor (MAb 43) and labelled with I-125-hGH, was used to quantitate plasma GHBP by Scatchard analysis in the developing (pooled plasma samples) and adult (individual animals) possums. Binding affinity (K-a) values in pouch young aged between 45 and 54 and 144 and 153 days post-partum varied between 1.0 and 2.4 x 10(9)/M, which was slightly higher than that in adult plasma (0.96 +/- 0.2 x 10(9)/M, n = 6). Binding capacity (B-max) values increased from non-detectable levels in animals aged 25-38 days post-partum to reach concentrations around half that seen in the adult (1.4 +/- 0.2 x 10(-9) M) by about 117 days post-partum and remained at this level until 153 days post-partum. Therefore, in early pouch life when plasma GH concentrations are highest, the very low concentrations of GHBP are unlikely to be important in terms of competing with GH-receptor for ligand or altering the half-life of circulating GH.

Application of GA3 and girdling of branches on the production of extemporaneous fruits of 'Tahiti' acid lime

The objective of this study was to evaluate the effect of girdling of branches and the application of GA3, still in the main crop flowering on the formation of reproductive structures, the fruit set of the following extemporaneous flowering of 'Tahiti' acid lime. This work used a randomized block design with five replications in a 4x3 factorial design. The first factor was the foliar application of GA3, 88 days before extemporaneous flowering at five concentrations (0, 7, 14 and 21 mg.L-1). The second factor was the girdling times, 108 days before flowering onset (A108AF), 78 days before flowering (A78AF) plus no-girdling control (NG). In extemporaneous flowering, the proportion of buds, flowers and fruits that abscised was 38.75, 31.25 and 18.64% respectively and 11.34% of all reproductive structures formed were collected. GA3 did not affect fruit set. Girdling increased fruit set by 229 and 256% at A108DAF and A78DAF respectively, in relation to NG plants. The GA3 reduced the formation of floral structures, which resulted in a quadratic increase in field production by reducing the competition for photoassimilates among fruits. The concentration of 21 mg.L-1 was the most productive, with 15.97 fruit.plant-1.

Multi-scaled femtosecond laser structuring of stationary titanium surfaces

The evolution of the topography of titanium surfaces treated with femtosecond laser radiation in stationary conditions as a function of radiation fluence and number of laser pulses is investigated. Depending on the processing parameters, ripples, microcolumns, wavy or smooth surfaces can be obtained. The ripples predominate for fluences near the damage threshold of titanium (0.2+/-0.1) J/cm(2), while microcolumns form during the first 200 pulses for fluences between (0.6+/-0.2) and (1.7+/-0.2) J/cm(2). A wavy topography develops for fluences and number of pulses higher than (1.7+/-0.2) J/cm(2) and 300, respectively. A bimodal surface topography consisting of surface ripples overlapping a microcolumnar topography can be obtained if the surfaces are firstly treated to create microcolumns followed by laser treatment with a lower fluence near the ablation threshold of the material, in order to generate periodic ripple

Layer-by-layer immobilization of carbon dots fluorescent nanomaterials on single optical fiber

We report within this paper the development of a fiber-optic based sensor for Hg(II) ions. Fluorescent carbon nanoparticles were synthesized by laser ablation and functionalized with PEG200 and N-acetyl-l-cysteine so they can be anionic in nature. This characteristic facilitated their deposition by the layer-by-layer assembly method into thin alternating films along with a cationic polyelectrolyte, poly(ethyleneimine). Such films could be immobilized onto the tip of a glass optical fiber, allowing the construction of an optical fluorescence sensor. When immobilized on the fiber-optic tip, the resultant sensor was capable of selectively detecting sub-micromolar concentrations of Hg(II) with an increased sensitivity compared to carbon dot solutions. The fluorescence of the carbon dots was quenched by up to 44% by Hg(II) ions and interference from other metal ions was minimal.

Biodiesel production over lithium modified lime catalysts: activity and deactivation

Biodiesel production by methanolysis of semi-refined rapeseed oil was studied over lime based catalysts. In order to improve the catalysts basicity a commercial CaO material was impregnated with aqueous solution of lithium nitrate (Li/Ca = 03 atomic ratio). The catalysts were calcined at 575 degrees C and 800 degrees C, for 5 h, to remove nitrate ions before reaction. The XRD patterns of the fresh catalysts, including the bare CaO, showed lines ascribable to CaO and Ca(OH)(2). The absence of XRD lines belonging to Li phases confirms the efficient dispersion of Li over CaO. In the tested condition (W-cat/W-oil = 5%; CH3OH/oil = 12 molar ratio) all the fresh catalysts provided similar biodiesel yields (FAME >93% after 4 h) but the bare CaO catalyst was more stable. The activity decay of the Li modified samples can be related to the enhanced, by the higher basicity, calcium diglyceroxide formation during methanolysis which promotes calcium leaching. The calcination temperature for Li modified catalysts plays an important role since encourages the crystals sinterization which appears to improve the catalyst stability. (C) 2013 Elsevier B.V. All rights reserved.