Protective effect of Calendula officinalis extract against UVB-induced oxidative stress in skin: Evaluation of reduced glutathione levels and matrix metalloproteinase secretion

Background and purpose: Calendula officinalis flowers have long been employed time in folk therapy, and more than 35 properties have been attributed to decoctions and tinctures from the flowers. The main uses are as remedies for burns (including sunburns), bruises and cutaneous and internal inflammatory diseases of several origins. The recommended doses are a function both of the type and severity of the condition to be treated and the individual condition of each patient. Therefore, the present study investigated the potential use of Calendula officinalis extract to prevent UV irradiation-induced oxidative stress in skin. Methods: Firstly, the physico-chemical composition of marigold extract(ME) (hydroalcoholic extract)was assessed and the in vitro antioxidant efficacy was determined using different methodologies. Secondly, the cytotoxicity was evaluated in L929 and HepG2 cells with the MTT assay. Finally, the in vivo protective effect of ME against UVB-induced oxidative stress in the skin of hairless mice was evaluated by determining reduced glutathione (GSH) levels and monitoring the secretion/activity of metalloproteinases. Results and conclusions: The polyphenol, flavonoid, rutin and narcissin contents found in ME were 28.6 mg/g, 18.8 mg/g, 1.6 mg/g and 12.2 mg/g, respectively and evaluation of the in vitro antioxidant activity demonstrated a dose-dependent effect of ME against different radicals. Cytoxicity experiments demonstrated that ME was not cytotoxic for L929 and HepG2 cells at concentrations less than or equal to of 15 mg/mL However, concentrations greater than or equal to 30 mg/mL, toxic effects were observed. Finally, oral treatment of hairless mice with 150 and 300 mg/kg of ME maintained GSH levels close to non-irradiated control mice. In addition, this extract affects the activity/secretion of matrix metalloproteinases 2 and 9 (MMP-2 and -9) stimulated by exposure to UVB irradiation. However, additional studies are required to have a complete understanding of the protective effects of ME for skin. (C) 2009 Elsevier Ireland Ltd. All rights reserved.

Pre-treatment of cattle sperm and/or oocyte with antibody to lipocalin type prostaglandin D synthase inhibits in vitro fertilization and increases sperm-oocyte binding

The present study was conducted to determine the affect of pre-treating of oocytes and/or sperm with a rabbit polyclonal antibody against recombinant cattle lipocalin type prostaglandin D synthase (alpha L-PGDS) on in vitro sperm-oocyte binding and fertilization. In vitro matured cattle oocytes were incubated (39 degrees C, 5% CO2 in air) for I It in the following treatments either 500 mu L of fertilization medium (FM) or FM with alpha L-PGDS (1:2000). Frozen-thawed spermatozoa were washed by a 45/90% layered Percoll gradient centrifugation and incubated for I h either FM or FM with a L-PGDS. This study utilized five different treatments: (1) no antibody (control); (2) a rabbit IgG against a non-bovine antigen, bacterial histidase (alpha-hist); (3) a L-PGDS at fertilization time (with fertilization medium); (4) alpha L-PGDS-treated oocytes; or (5) a L-PGDS-treated sperm. Pre-treated oocytes were incubated with 10 X 10(4) washed spermatozoa per 25 oocytes. Oocytes used to assess sperm binding were stained with Hoescht 33342, and the number of sperm bound per zonae pellucidae counted. The remaining oocytes were fixed in acid alcohol, stained with 1% acetate-orcein and observed to determine the presence of pronuclei. More sperm bound to the zonae pellucidae when oocytes and/or sperm were pre-treated with alpha. L-PGDS: (1) 26.4 +/- 3.0; (2) 25.6 +/- 3.0; (3) 59.7 +/- 3.0; (4) 56.4 +/- 3.0; and (5) 57.1 +/- 3.0. Addition of alpha L-PGDS with sperm, oocytes, or both, decreased fertilization (P < 0.05) compared with the control: (1) 89.2 +/- 2.0%; (2) 87.5 +/- 2.0%; (3) 19.4 +/- 2.0%; (4) 27.2 +/- 3.1%; and (5) 14.1 +/- 3.4%. The alpha L-PGDS reacts with both oocytes and spermatozoa, resulting in increases of in vitro sperm-oocyte binding and inhibition of fertilization. These observations suggest that L-PGDS may have a role in cattle fertilization. (c) 2007 Elsevier B.V. All rights reserved.

Dentine caries inhibition through CO(2) laser (10.6 mu m) irradiation and fluoride application, in vitro

Objective: The purpose of the study was to investigate whether dentine irradiation with a pulsed CO(2) laser (10.6 mu m) emitting pulses of 10 ms is capable of reducing dentine calcium and phosphorus losses in an artificial caries model. Design: The 90 dentine slabs obtained from bovine teeth were randomly divided into six groups (n = 15): negative control group (GC); positive control group, treated with fluoride 1.23% (GF); and laser groups irradiated with 8 J/cm(2) (L8); irradiated as in L8 + fluoride 1.23% (L8F); irradiated with 11j/cm(2) (L11); irradiated as in L11 + fluoride 1.23% (L11F). After laser irradiation the samples were submitted to a pH-cycling model for 9 days. The calcium and phosphorous contents in the de- and remineralization solutions were measured by means of inductively coupled plasma optical emission spectrometer - ICP-OES. Additionally intra-pulpal temperature measurements were performed. The obtained data were analysed by means of ANOVA and Tukey`s test (alpha = 0.05). Results: In the demineralization solutions the groups L11F and GF presented significantly lower means of calcium and phosphorous losses than the control group; and in L11F means were significantly lower than in the fluoride group. Both irradiation parameters tested caused intrapulpal temperature increase below 2 degrees C. Conclusion: It can be concluded that under the conditions of this study, CO(2) laser irradiation (10.6 mu m) with 11J/cm(2) (540 mJ and 10 Hz) of fluoride treated dentine surfaces decreases the loss of calcium and phosphorous in the demineralization process and does not cause excessive temperature increase inside the pulp chamber. (C) 2010 Elsevier Ltd. All rights reserved.

Influence of different volumes of EDTA for final rinse on smear layer removal

Objective. The aim of this study was to analyze the influence of different volumes of 17% EDTA for final rinse on smear layer removal on the different areas of the root canal. Study design. Forty single-rooted teeth were instrumented using rotary instruments. The teeth were divided into 3 test groups according to the EDTA volume for final rinse (5 mL, 10 mL, 15 mL) and 1 control group (10 mL of 1% sodium hypochlorite). The roots were axially split into halves, and the smear layer removal from the canals was determined under scanning electron microscope. The data were analyzed using Kruskal Wallis and Dunn tests. Results. The 3 experimental groups showed no statistical difference (P > .05); however, when the test groups were compared to the control group statistically significant differences were found (P < .01). The root canal wall surfaces of teeth in the control group showed the presence of heavy smear layer through the entire length of the root canals. The other groups showed mainly smear layer-free surfaces or a small amount of debris. When coronal, middle and apical thirds were compared, there was no statistically significant difference (P > .05). Conclusions. Based on the results, it appears that a final rinse with 5 mL of EDTA per canal provides good smear layer removal, with root canal walls free of debris and mostly open dentinal tubules in all areas. (Oral Surg Oral Med Oral Pathol Oral Radiol Endod 2008;106:e40-e43)

GAPD and tubulin are suitable internal controls for qPCR analysis of oral squamous cell carcinoma cell lines

The selection of housekeeping genes is critical for gene expression studies. To address this issue, four candidate housekeeping genes, including several commonly used ones, were investigated in oral squamous cell carcinoma cell lines. A simple quantitative RT-PCR approach was employed by comparing relative expression of the four candidate genes within two cancerous cell tines (HN6 and HN31) and one noncancerous cell tine (HaCaT) treated or not with EGF and TGF-beta 1. Data were analyzed using ANOVA followed by the NormFinder software program. On this basis, stability of the candidate housekeeping genes was ranked and non statistical differences were found using ANOVA test. On the other hand, the NormFinder was able to show that GAPD and TUBB presented the less variable results, representing appropriated housekeeping genes for the samples and conditions anatyzed. In conclusion, this study suggests that the GAPD and the TUBB represent adequate normalizers for gene profiting studies in OSCC cell. tines, covering, respectively, high and low expression levels genes. (C) 2008 Elsevier Ltd. All rights reserved.

Effect of the Combination of Sodium Hypochlorite and Chlorhexidine on Dentinal Permeability and Scanning Electron Microscopy Precipitate Observation

Introduction: This study compared the combined use of sodium hypochlorite (NaOCl) and chlorhexidine (CXH) with citric acid and CXH on dentinal permeability and precipitate formation. Methods: Thirty-four upper anterior teeth were prepared by rotary instrumentation and NaOCl. The root canal surfaces were conditioned for smear layer removal using 15% citric acid solution under ultrasonic activation and a final wash with distilled water. All teeth were dried, and 30 specimens were randomly divided into three equal groups as follows: positive control group (PC), no irrigation; 15% citric acid + 2% CHX group (CA + CHX); and 1% NaOCl + 2% CHX group (NaOCl + CHX). All roots were immersed in a 0.2% Rhodamine B solution for 24 hours. One-millimeter-thick slices from the cementum-enamel junction were scanned at 400 dpi and analyzed using the software ImageLab (LIDO-USP, Sao Paulo, Brazil) for the assessment of leakage in percentage. For scanning electron microscopy analysis, four teeth, irrigated for NaOCl + CHX samples, were split in half, and each third was evaluated at 1,000x and 5,000x (at the precipitate). Results: Using the analysis of variance test followed by the Bonferroni comparison method, no statistical differences between groups were found when analyzed at the cervical and medium thirds. At the apical third, differences between the PC and NaOCl + CHX (p<0.05) and CA + CHX and NaOCl + CHX could be seen (p < 0.05). Conclusion: The combination of 1% NaOCl and 2% CHX solutions results in the formation of a flocculate precipitate that acts as a chemical smear layer reducing the dentinal permeability in the apical third. (J Endod 2010;36:847-850)

Tooth abnormalities and soft tissue changes in patients with velocardiofacial syndrome

Objective. The objective of this study was to investigate the prevalence of tooth abnormalities and soft tissue changes in patients with velocardiofacial syndrome. Study design. Twenty-six patients with velocardiofacial syndrome were examined to investigate the presence of tooth abnormalities and soft tissue alterations. The occurrence of tooth agenesis and supernumerary teeth was compared to patients without morphofunctional alterations, matched for gender and age. Results. Of all patients, 76.92% exhibited at least one tooth abnormality, with predominance of hypoplastic alterations, especially represented by hypodevelopment of the lingual cusp of mandibular first premolars and enamel opacities. The occurrence of tooth agenesis and supernumerary teeth was similar in both study and control groups. Conclusion. the present results suggest an association between hypodevelopment of the lingual cusp of mandibular first premolars and enamel opacities, yet these findings still require corroboration. Future studies should further investigate these aspects in larger samples compared to control groups, as well as employing molecular genetics techniques.

Preventive effect of an iron varnish on bovine enamel erosion in vitro

Objective: The aim of this study was to evaluate, in vitro, the effect of an experimental varnish containing iron on the dissolution of bovine enamel by carbonated beverage. Methods: Eighty specimens were randomly allocated to four groups (n = 20 per group), according to the following treatments: Fe varnish (FeV, 10 mmoL/L Fe), F varnish (FV, 2.71% F), placebo varnish (PV) and control (not treated, NT). The varnishes were applied in a thin layer and removed after 6 h. Then, the samples were submitted to six cycles, alternating re- and demineralisation (only 1 day). Demineralisation was performed with the beverage Coca-Cola (R) (10 min, 30 mL/block) and remineralisation with artificial saliva for I h. In order to determine the amount of enamel dissolved, the wear was analysed by profilometry. Data were analysed by ANOVA and Tukey`s test (p < 0.05). Results: The mean wear (+/- S.E.) was significantly lesser for the FeV (0.451 +/- 0.018 mu m) when compared to the other treatments. The FV caused significantly less wear (0.554 +/- 0.022 mu m) when compared to PV (0.991 +/- 0.039 mu m) and NT (1.014 +/- 0.033), which did not significantly differ from each other. Conclusions: The results suggest that the iron varnish can interfere with the dissolution of dental enamel in the presence of acidic beverages. (C) 2008 Elsevier Ltd. All rights reserved.

Influence of root parallelism on the stability of extraction-site closures

Introduction: In premolar extraction cases, root parallelism is recommended to preserve the stability of space closures. The influence of the degree of root parallelism on relapse of tooth extraction spaces has been a controversial topic in the literature. The aim of this study was to compare the angle between the long axes of the canine and the second premolarin patients with and without stability of extraction-space closures. Methods: A sample of 56 patients, treated with 4 premolar extractions, was divided into 2 groups: group 1, consisting of 25 patients with reopening of extraction spaces; and group 2, consisting of 31 patients without reopening of extraction spaces. Panoramic radiographs of each patient were analyzed at the posttreatment and 1-year posttreatment stages. The data were statistically analyzed by using chi-square tests, t tests, analysis of variance (ANOVA), and Pearson correlation coefficients. Results: The results showed that the groups did not differ regarding the angle between the canine and the second premolar, and there was no correlation between angular changes and reopening of extraction spaces, showing that dental angular changes are not determining factors for relapse, and other factors should be investigated. Conclusions: The final angle and the posttreatment changes observed in the angle between the long axes of the canine and the second premolar showed no influence on the relapse of extraction spaces. (Am J Orthod Dentofacial Orthop 2011; 139: e505-e510)

Effects of accentuated and reversed curve of Spee on apical root resorption

Introduction: In this study, we evaluated the influence of intrusion mechanics with accentuated and reversed curve of Spee on root resorption of the maxillary and mandibular incisors. Methods: A sample of 60 patients with Class I and Class II Division 1 malocclusions having nonextraction treatment was divided into 2 groups with the following characteristics: group 1 comprised 30 deepbite patients, treated with accentuated and reversed curve of Spee intrusion mechanics, with an initial mean age of 12.8 +/- 1.23 years (range, 10.01-15.32 years), and group 2 comprised 30 patients with normal overbite treated without intrusion mechanics, with an initial mean age of 12.87 +/- 1.43 years ( range, 10.02-15.36 years). Pretreatment and posttreatment periapical radiographs were used to evaluate root resorption. The groups were compared by using the Mann-Whitney U test. Correlation between root resorption and tooth movement was investigated with the Spearman correlation coefficient. Results: The deepbite group treated with accentuated and reversed curve of Spee had statistically greater root resorption ( 1.87) than the normal overbite group ( 1.54), at P=.017. Changes in overbite and vertical displacements of the maxillary central incisor apices had significant correlations to root resorption ( r = 0.30, P =.019; r = 0.27, P =.037, respectively). Conclusions: Accentuating and reversing the curve of Spee in the archwires to correct deep overbite causes more root resorption than nonintrusive mechanics.

Cleft Palate Obturation With Branemark Protocol Implant-Supported Fixed Denture and Removable Obturator

A 41-year-old man with cleft palate presented with a wide dehiscence and missing teeth. Six implants had been placed for fabrication of an overdenture, which was unsatisfactory. A bar was waxed and cast for connection to the implants; precision attachments were placed laterally for retention. A fixed partial denture was fabricated, and milled crowns were fabricated at the molar region to provide a guiding plane for insertion of a removable palatal obturator. Good swallowing and speech outcomes were achieved. This technique provided functional and esthetic benefits, enhanced oral hygiene, and improved the psychological condition of the patient.

Post space cleaning using a new nickel titanium endodontic drill combined with different cleaning regimens

This study compared the effect of two drills and five cleaning regimens on post space debridement. One hundred extracted premolars were instrumented and obturated with warm vertical compaction of gutta percha. The teeth were divided into two groups according to the drill used to remove gutta percha/sealer and for post space preparation: a Largo drill (Largo; Dentsply, St Quentin en Yvelines, France) or a MTwo-PF drill (Sweden&Martina, Due Carrare, Padova, Italy). The following cleaning regimens were used: EDTA, ultrasonics, ultrasonics + EDTA, phosphoric acid, and distilled water. Scanning electron microscopic images of the post spaces were taken, and the presence of debris and of open dentin tubules were evaluated. The ultrasonics + EDTA, phosphoric acid, and EDTA groups were comparable in open tubules scores for both drills and in debris scores after the use of MTwo-PF (p > 0.05). The ultrasonics and control groups performed significantly worse (p < 0.05). The MTwo-PF drill resulted as effective as the Largo drill in obtaining a good post space cleaning, especially when followed by ultrasonics + EDTA irrigant regimen.

Central NOS inhibition differentially affects vasopressin gene expression in hypothalamic nuclei in septic rats

Our aim was to investigate the effect of central NOS inhibition on hypothalamic arginine vasopressin (AVP) gene expression, hormone release and on the cardiovascular response during experimental sepsis. Male Wistar rats were intracerebroventricularly injected with the non-selective NO synthase (NOS) inhibitor (L-NAME) or aminoguanidine, a selective inhibitor of the inducible isoform (iNOS). After 30 min. sepsis was induced by cecal ligation and puncture (CLP) causing an increase in heart rate (HR), as well as a reduction in median arterial pressure (MAP) and AVP expression ratio (AVP(R)), mainly in the supraoptic nucleus. AVP plasma levels (AVP(P)) increased in the early but not in the late phase of sepsis. L-NAME pretreatment increased MAP but did not change HR. It also resulted in an increase in AVP(P) at all time points, except 24 h, when it returned to basal levels. AVP(R), however remained reduced in both nuclei. Aminoguanidine pretreatment resulted in increased MAP in the early phase and higher AVP(R) in the supraoptic, but not in the paraventricular nucleus, while AVP(P) remained elevated at all time points. We suggest that increased central NO production, mainly inducible NOS-derived, reduces AVP gene expression differentially in supraoptic and paraventricular nuclei, and that this may contribute to low AVP plasma levels and hypotension in the late phase of sepsis. (c) 2010 Elsevier B.V. All rights reserved.

Eag1 potassium channel immunohistochemistry in the CNS of adult rat and selected regions of human brain

Eag1 (K(v)10.1) is the founding member of an evolutionarily conserved superfamily of voltage-gated K+ channels. In rats and humans Eag1 is preferentially expressed in adult brain but its regional distribution has only been studied at mRNA level and only in the rat at high resolution. The main aim of the present study is to describe the distribution of Eag1 protein in adult rat brain in comparison to selected regions of the human adult brain. The distribution of Eag1 protein was assessed using alkaline-phosphatase based immunohistochemistry. Eag1 immunoreactivity was widespread, although selective, throughout rat brain, especially noticeable in the perinuclear space of cells and proximal regions of the extensions, both in rat and human brain. To relate the results to the relative abundance of Eag1 transcripts in different regions of rat brain a reverse-transcription coupled to quantitative polymerase chain reaction (real time PCR) was performed. This real time PCR analysis showed high Eag1 expression in the olfactory bulb, cerebral cortex, hippocampus, hypothalamus, and cerebellum. The results indicate that Eag1 protein expression greatly overlaps with mRNA distribution in rats and humans. The physiological relevance of potassium channels in the different regions expressing Eag1 protein is discussed. (C) 2008 IBRO. Published by Elsevier Ltd. All rights reserved.

The impact of nutrition support on body composition in cancer outpatients receiving radiotherapy

This study investigated the change in body composition in 36 cancer outpatients receiving radiotherapy to the head and neck area (mean age: 63 ± 15 years) randomised to receive either nutrition intervention (NI; n=15) or usual care (UC; n=21). Body weight and composition were measured at the commencement of radiotherapy and 3 months later. The UC group lost significantly more weight; mean decrease = 4.3 kg, than the NI group: mean decrease = 1.1 kg (t(30)=-2.5, p=0.019). Fat-free mass loss was significantly higher in the UC group with a mean loss of 2.2 kg versus 0.3 kg in the NI group (t(30)=- 2.3, p=0.029). Body composition as measured by foot-to-foot bioelectrical impedance analysis provides more information than weight alone and can allow for tailoring of NI.