Dunn's Multiple Comparison Test Research Articles

The bleaching efficiency of KTP and diode 810 nm lasers on teeth stained with different substances: An in vitro study.

The purpose of this study was to evaluate the bleaching efficiency of two different lasers (KTP and diode 810 nm) on teeth, randomly divided by means an Excel function (Microsoft Excel 2010 "Fx causale") and stored in physiological solution, that were previously stained with different substances commonly considered as a cause of tooth discoloration, such as coffee, tea and red fruits and to investigate the role of laser irradiation in an experimental model, during the dental bleaching process. Three groups of 45 bovine teeth were created and immersed for one week in a solution of tea, coffee or red fruits respectively. Each group was divided into three sub-groups of fifteen teeth. One was bleached with a 30% hydrogen peroxide gel for 30 min only as control, another 15 teeth group was bleached with the gel plus 810 nm diode laser irradiation and the last group was bleached with the gel plus KTP irradiation. The lasers were applied in three cycles of 30 sec each with a power of 1.5 W localized on a 10 mm spot on the teeth. The temperature of the gel was checked during the bleaching procedure using a thermometer and the colour of each tooth was measured by a spectrophotometer. Statistical analysis of the collected data was performed using Graph Pad Prism, version 6.01 software, Kruskal-Wallis test and Dunn's multiple comparison test and Mann-Whitney test. P value <0.0001 was considered extremely significant (***), P value between 0.001 to 0.01 very significant (**), P value between 0.01 to 0.05 significant (*) and P value >0.05 not significant (ns). By these tests diode laser was effective only at bleaching teeth stained with coffee meanwhile the KTP laser was efficient at bleaching teeth with coffee, tea and red fruits stains. This study suggests that a relation between the laser wavelength and the type of staining on the dental enamel and the efficacy of the whitening treatment exists.

Seventy-two hours of mild hypothermia after cardiac arrest is associated with a lowered inflammatory response during rewarming in a prospective observational study

Whole-body ischemia and reperfusion trigger a systemic inflammatory response. In this study, we analyzed the effect of temperature on the inflammatory response in patients treated with prolonged mild hypothermia after cardiac arrest. Ten comatose patients with return of spontaneous circulation after pulseless electrical activity/asystole or prolonged ventricular fibrillation were treated with mild therapeutic hypothermia for 72 hours after admission to a tertiary care university hospital. At admission and at 12, 24, 36, 48, 72, 96 and 114 hours, the patients' temperature was measured and blood samples were taken from the arterial catheter. Proinflammatory interleukin 6 (IL-6) and anti-inflammatory (IL-10) cytokines and chemokines (IL-8 and monocyte chemotactic protein 1), intercellular adhesion molecule 1 and complement activation products (C1r-C1s-C1inhibitor, C4bc, C3bPBb, C3bc and terminal complement complex) were measured. Changes over time were analyzed with the repeated measures test for nonparametric data. Dunn's multiple comparisons test was used for comparison of individual time points. The median temperature at the start of the study was 34.3°C (33.4°C to 35.2°C) and was maintained between 32°C and 34°C for 72 hours. All patients were passively rewarmed after 72 hours, from (median (IQR)) 33.7°C (33.1°C to 33.9°C) at 72 hours to 38.0°C (37.5°C to 38.1°C) at 114 hours (P <0.001). In general, the cytokines and chemokines remained stable during hypothermia and decreased during rewarming, whereas complement activation was suppressed during the whole hypothermia period and increased modestly during rewarming. Prolonged hypothermia may blunt the inflammatory response after rewarming in patients after cardiac arrest. Complement activation was low during the whole hypothermia period, indicating that complement activation is also highly temperature-sensitive in vivo. Because inflammation is a strong mediator of secondary brain injury, a blunted proinflammatory response after rewarming may be beneficial.

Dental bleaching with a 10% hydrogen peroxide product: A six-month clinical observation

The possibility of bleaching vital teeth with peroxide-based products considerably revolutionized esthetic dentistry. The aim of this clinical study was to evaluate tooth color change and dental sensitivity after exposure to preloaded film containing a 10% hydrogen peroxide whitening system (Opalescence Trθswhite Supreme). A total of 13 volunteers, aged 18 to 25 years, participated in this study. The patients used the whitening system once a day for 60 minutes during the 8-day study. For maxillary incisors and canines, the color change was visually evaluated with the Vita color scale before, immediately, and six months after the treatment. Tooth sensitivity was evaluated during the daily gel applications. All whitening applications were done in office and under the supervision of a dental professional. The results were analyzed using the Friedman Test (nonparametric repeated measures ANOVA) at a level of 5%, and Dunn's Multiple Comparison Test at the level of 5%. It was verified that the original mean color values observed at the baseline analysis differed significantly from those observed immediately after bleaching, as well as from those seen in the analysis at six months ( P = 0.001). There was no significant difference between the mean color values observed in the immediate time and in the analysis at six months ( P = 0.474). No tooth sensitivity was observed in any patients. It was concluded that the bleaching technique using the 10% hydrogen peroxide system was effective in a short period of time without tooth sensitivity during applications.

Oxidative stress and labile plasmatic iron in anemic patients following blood therapy.

To determine the plasmatic iron content and evaluate the oxidative stress (OS) markers in subjects receiving blood therapy. Thirty-nine individuals with unspecified anemia receiving blood transfusions and 15 healthy subjects were included in the study. Anemic subjects were divided into three subgrouP: (1) those that received up to five blood transfusions (n = 14); (2) those that received from five to ten transfusions (n = 11); and (3) those that received more than ten transfusions (n = 14). Blood samples were collected by venous arm puncture and stored in tubes containing heparin. The plasma and cells were separated by centrifugation and subsequently used for analyses. Statistical analyses were performed using Kruskal-Wallis analysis of variance followed by Dunn's multiple comparison tests when appropriate. The eletrophoretic hemoglobin profiles of the subjects included in this study indicated that no patients presented with hemoglobinopathy. Labile plasmatic iron, ferritin, protein carbonyl, thiobarbituric acid-reactive substances (TBARS) and dichlorofluorescein diacetate oxidation were significantly higher (P < 0.05), whereas total thiol levels were significantly lower (P < 0.05) in transfused subjects compared to controls. Additionally, the activity of catalase, superoxide dismutase and glutathione peroxidase were significantly lower in the transfused subjects (P < 0.05). Antioxidant enzyme activities and total thiol levels were positively correlated (P < 0.05), and negatively correlated with the levels of protein carbonyl and TBARS (P < 0.05). In contrast, protein carbonyl and TBARS were positively correlated (P < 0.05). Altogether, these data confirm the involvement of OS in patients following therapy with repeated blood transfusions. Our data reveal that changes in OS markers are correlated with levels of labile plasmatic iron and ferritin and the number of transfusions.

Endocrine cells in the oxyntic mucosa of the stomach in patients with irritable bowel syndrome.

To study the different endocrine cell types in the oxyntic mucosa of patients with irritable bowel syndrome (IBS). Seventy-six patients with IBS were included in the study (62 females and 14 males; mean age 32 years, range 18-55 years), of which 40 also fulfilled the Rome III criteria for functional dyspepsia (FDP). Of the entire IBS cohort, 26 had diarrhea as the predominant symptom (IBS-D), 21 had a mixture of diarrhea and constipation (IBS-M), and 29 had constipation as the predominant symptom (IBS-C). Forty-three age and sex-matched healthy volunteers without any gastrointestinal complaints served as controls. The patients were asked to complete the Birmingham IBS symptom questionnaire. Both the patients and controls underwent a standard gastroscopy, during which three biopsy samples were taken from the corpus. Sections from these biopsy samples were immunostained using the avidin-biotin complex (ABC) method, for ghrelin, serotonin, somatostatin and histamine. The densities of these cell types and immunoreactivity intensities were quantified using computerized image analysis with Olympus cellSens imaging software (version 1.7). The densities of the ghrelin cells in the control, IBS-total, IBS-D, IBS-M and IBS-C groups were 389 (320, 771), 359 (130, 966), 966 (529, 1154), 358 (120, 966) and 126 (0, 262) cells/mm(2), respectively. There was a significant difference between the tested groups (P < 0.0001). Dunn's multiple comparison test showed that the ghrelin cell density was significantly higher in IBS-D and lower in IBS-C than in the controls (P = 0.03 and 0.0008, respectively). The ghrelin cell density in patients with both IBS and FDP was 489 (130, 966), and in those with IBS only 490 (130, 956). There was no statistical significant difference between these 2 groups of patients (P = 0.9). The immunoreactivity intensity did not differ between any of the groups (P = 0.6). The diarrhea score of the Birmingham IBS symptom questionnaire was significantly positively correlated with ghrelin cell density (r = 0.65; P < 0.0001) and significantly inversely correlated with that of constipation (r = 90.69; P < 0.0001). The densities of the serotonin cells were 63 (51, 82), 51 (25, 115), 120 (69, 128), 74 (46, 123) and 40 (0, 46) cells/mm(2) in the control, IBS-total, IBS-D, IBS-M and IBS-C groups, respectively. A statistically significant difference was found between the tested groups (P < 0.0001). Posttest revealed that serotonin cell density was significantly higher in IBS-D and lower in IBS-C than in controls (P = 0.02 and 0.004, respectively), but did not differ in the IBS-total and IBS-M groups from that in controls (P = 0.5 and 0.4, respectively). The serotonin cell density in patients with both IBS and FDP was 62 (25, 115) and in those with IBS only 65 (25, 123). There was no statistically significant difference between these 2 groups of patients (P = 1). The immunoreactivity intensity of serotonin did not differ significantly between any of the groups (P = 0.0.9). The serotonin cell density was significantly positively correlated with the diarrhea score of the Birmingham IBS symptom questionnaire (r = 0.56; P < 0.0001) and significantly inversely correlated with that of constipation (r = 0.51; P < 0.0001). The densities of the somatostatin cells were 97 (72, 126), 72 (0, 206), 29 (0, 80), 46 (0, 103) and 206 (194, 314) cells/mm(2) in the control, IBS-total, IBS-D, IBS-M and IBS-C groups, respectively (Figures 7 and 8). There was a statistically significant difference between the controls and the IBS subgroups (P < 0.0001). The density of somatostatin cells was significantly lower in the IBS-D and IBS-M groups but higher in IBS-C patients than in the controls (P < 0.01, P = 0.02, and P = 0.0008, respectively). The somatostatin cell density in patients with both IBS and FDP was 86 (0-194), and in those with IBS only 110 (0-206). There was no statistically significant difference between these 2 groups of patients (P = 0.6). There was no significant difference in somatostatin immunoreactivity intensity between the controls. The diarrhea score of the Birmingham IBS symptom questionnaire was inversely correlated with somatostatin cell density (r = 0.38; P = 0.0007) and was positively correlated with that of constipation (r = 0.64; P < 0.0001). The finding of abnormal endocrine cells in the oxyntic mucosa shows that the endocrine cell disturbances in IBS are not restricted to the intestine. Furthermore, it appears that ghrelin, serotonin and somatostatin in the oxyntic mucosa of the stomach may play an important role in the changing stool habits in IBS through their effects on intestinal motility.

Effects of variations of osmolarity on behavior in Leptodactylus macrosternum Miranda-Ribeiro, 1926 (Leptodactylidae, Anura, Amphibia)

Amphibians experiencing evapotranspirative water loss may suffer dehydration. Another difficulty is their limited tolerance to brackish water. This study aimed to investigate the effect of water salinity on behavior (activity and water conservation posture) in Leptodactylus macrosternum, in normally hydrated animals with urine in their bladder, in normally hydrated animals without urine in their bladder and in dehydrated ones with urine removed from their bladder. Experiments were filmed, behaviors were registered, and a Behavioral Dehydration Protection Index (BDPI) was calculated, using a weighted average of the postures. A GLM and Kruskall Wallis test was performed with Dunn's multiple comparisons test to evaluate the effects of osmotic gradient stress on BDPI. It was found that emptied bladder and aerial dehydration did not influence BDPI (p = 0.832 and p = 0.142, respectively), contrary to what is seen in the literature. The different osmotic media had a significant effect on BDPI (p = 0.0003).

Pulse Volume Recording Does Not Enhance Segmental Pressure Readings for Peripheral Arterial Disease Stratification

Noninvasive vascular laboratory determinations for peripheral arterial disease (PAD) often combine pulse volume recordings (PVRs), segmental pressure readings (SPs), and Doppler waveform traces (DWs) into a single diagnostic report. Our objective was to assess the corresponding diagnostic values for each test when subjected to interpretation by 4 vascular specialists. A total of 2226 non-invasive diagnostic reports were reviewed through our institutional database between January 2009 and December 2011. Data from noninvasive records with corresponding angiograms performed within 3 months led to a cohort of 76 patients (89 limbs) for analysis. Four vascular specialists, blinded to the angiographic results, stratified the noninvasive studies as representative of normal, <50% "subcritical," or ≥50% "critical" stenosis at the upper thigh, lower thigh, popliteal, and calf segments using 4 randomized noninvasive modalities: (1) PVR alone; (2) SP alone; (3) SP+DW; and (4) SP+DW+PVR. The angiographic records were independently graded by another 3 evaluators and used as a standard to determine the noninvasive diagnostic values and interobserver agreements for each modality. Statistical tests used include the Fleiss-modified kappa analysis, Kruskal-Wallis analysis of variance with Dunn's multiple comparison test, the Kolmogorov-Smirnov test, and the unpaired t-test with Welch's correction. Interobserver variance for all modalities was high, except for SP. When surveying for any stenosis (<50% and ≥50%), sensitivity (range 25-75%) was lower than specificity (range 50-84%) for all modalities. When surveying for critical stenosis only (≥50%), sensitivity (range 27-54%) was also lower than specificity (range 68-92%). Accuracy for detecting any stenosis with SP+DW was significantly higher than with PVR alone (66 ± 7% vs. 56 ± 12%, P = 0.017). There was a significant reduction in accuracy when including incompressible readings within the SP-only analysis compared with exclusion of incompressible vessels (P = 0.0006). However, the effect of vessel incompressibility on accuracy was removed with the addition of DW (P = 0.17) to the protocol. SP has the greatest interobserver agreement in evaluation of PAD and can be used preferentially for PAD stratification. Given the lower accuracy of PVR for detecting either subcritical or critical disease, PVR tests can be omitted from the noninvasive vascular examination without a significant reduction in overall diagnostic value and can be reserved for patients with incompressible vessels.

Platelet-Rich Plasma: Quantitative Assessment of Growth Factor Levels and Comparative Analysis of Activated and Inactivated Groups

BackgroundPlatelet-rich plasma (PRP) has more concentrated platelets than normal plasma (approximately 150-400×103 cell/dL). Platelets excrete several growth factors and cytokines that are associated with the healing and regeneration process. However, even though PRP is widely used, the mechanism or actual effect is presently unclear. Therefore, this study was performed to investigate the levels of growth factors and platelet concentration rate.MethodsAutologous blood for preparing PRP was obtained from healthy subjects aged 25 to 35 years. The samples were divided into 4 experimental groups (inactivated whole blood, inactivated PRP, activated whole blood with thrombin and calcium chloride, and activated PRP). The platelet counts in the blood were analyzed and the growth factors were quantitatively measured. A statistical analysis was performed by using Dunn's multiple comparison test.ResultsIn the blood cell analysis, the platelet count of the PRP group was approximately 4.25 times higher than that of the whole blood group. In the quantitative analysis of growth factors, the platelet-derived growth factor (PDGF)-AB, PDGF-BB, and transforming growth factor-β of the inactivated and activated PRP groups were higher than those of the inactivated and activated whole blood groups (P<0.05).ConclusionsIn this study, the platelet count and the levels of PDGF-AB and PDGF-BB in the PRP were determined. Further, more research is required on the bioactivity level of the growth factors secreted during the process of PRP preparation and the potency of growth factors that can be exerted physiologically in vivo.

The expression and immunohistochemical localization of periostin in odontogenic tumors of mixed epithelial/mesenchymal origin

The object of this study was to determine the expression and localization of periostin in the major mixed odontogenic tumors and to correlate any differential staining of the mesenchymal components to the interrelationship of these tumors. Five ameloblastic fibromas, 8 ameloblastic fibro-odontomas and 10 odontomas were assessed immunohistochemically for periostin staining. Because mesenchymal tissues were consistently present in all studied cases, these tissues were selected for statistical analysis of differential periostin staining. Periostin was variably localized to the mesenchymal component of the tumors as well as to preameloblasts and ameloblasts. Analysis of the mesenchymal staining intensity was statistically significantly different between ameloblastic fibro-odontomas and odontomas (P < .001; Dunn multiple comparisons test). Our results document periostin staining in human mixed odontogenic tumors. Statistical analysis of differential stromal staining supports the concept that the ameloblastic fibroma is a histogenetically distinct neoplasm as compared to ameloblastic fibro-odontoma and odontoma.

Effect of dexmedetomidine, morphine-lidocaine-ketamine, and dexmedetomidine-morphine-lidocaine-ketamine constant rate infusions on the minimum alveolar concentration of isoflurane and bispectral index in dogs

To determine the effect of dexmedetomidine, morphine-lidocaine-ketamine (MLK), and dexmedetomidine-morphine-lidocaine-ketamine (DMLK) constant rate infusions on the minimum alveolar concentration (MAC) of isoflurane and bispectral index (BIS) in dogs. 6 healthy adult dogs. Each dog was anesthetized 4 times with a 7-day washout period between anesthetic episodes. During the first anesthetic episode, the MAC of isoflurane (baseline) was established. During the 3 subsequent anesthetic episodes, the MAC of isoflurane was determined following constant rate infusion of dexmedetomidine (0.5 μg/kg/h), MLK (morphine, 0.2 mg/kg/h; lidocaine, 3 mg/kg/h; and ketamine, 0.6 mg/kg/h), or DMLK (dexmedetomidine, 0.5 μg/kg/h; morphine, 0.2 mg/kg/h; lidocaine, 3 mg/kg/h; and ketamine 0.6 mg/kg/h). Among treatments, MAC of isoflurane was compared by means of a Friedman test with Conover posttest comparisons, and heart rate, direct arterial pressures, cardiac output, body temperature, inspired and expired gas concentrations, arterial blood gas values, and BIS were compared with repeated-measures ANOVA and a Dunn test for multiple comparisons. Infusion of dexmedetomidine, MLK, and DMLK decreased the MAC of isoflurane from baseline by 30%, 55%, and 90%, respectively. Mean heart rates during dexmedetomidine and DMLK treatments was lower than that during MLK treatment. Compared with baseline values, mean heart rate decreased for all treatments, arterial pressure increased for the DMLK treatment, cardiac output decreased for the dexmedetomidine treatment, and BIS increased for the MLK and DMLK treatments. Time to extubation and sternal recumbency did not differ among treatments. Infusion of dexmedetomidine, MLK, or DMLK reduced the MAC of isoflurane in dogs.

The inhibitory effect of proanthocyanidin on soluble and collagen-bound proteases

ObjectiveThis study evaluated the inhibitory effect of proanthocyanidin (PA), a natural collagen cross-linker, on soluble and matrix-bound proteases, which are responsible for progressive degradation of exposed collagen fibrils within the hybrid layer and resin–dentine bond failure over time. MethodsThe inhibitory effects of PA (1%, 2%, 3%, 4.5% and 6%) on soluble recombinant matrix metalloproteinases (MMP-2, -8 and -9) and cysteine cathepsins (cathepsin B and K) were evaluated using MMP and cysteine cathepsins fluorometric assay kits. Chlorhexidine (CHX) was used as an inhibitor control. The effect of PA on endogenous matrix-bound proteases was examined by determining the change in dry mass of demineralized dentine beams and solubilized collagen peptides over 30 days. Two-way ANOVA and Tukey multiple comparison tests were used to analyze the effect of PA and proteases on the percentage inhibition of soluble proteases (α=0.05). Kruskal–Wallis one-way ANOVA and Dunn's multiple comparison tests were used to analyse the effect of PA on loss of dry mass and hydroxyproline content over time (α=0.05). ResultsProanthocyanidin inactivated more than 90% of soluble recombinant MMP-2, -8 and -9 and around 75–90% of cysteine cathepsin B and K, which was significantly higher than CHX (P<0.05). The inhibition of endogenous proteases by PA increased in a dose-dependent manner. The loss of dry mass and hydroxyproline release in the medium over time was the lowest in dentine beams pretreated with PA<CHX<control (P<0.05). ConclusionProanthocyanidin exhibited both dentine MMP and cysteine cathepsins inhibition, which was higher than chlorhexidine.

Obesity and worsening of chronic venous disease and joint mobility

The aim of this study was to investigate a possible relationship between obesity and decreased mobility of the talocrural joint and in turn chronic venous disease. One hundred obese patients recruited at Hospital Santa Casa de Maringa, Parana were enrolled by order of arrival at the hospital in a randomized quantitative cross-sectional study. Inclusion criteria were patients with a body mass index above 30 kg/m(2) and the exclusion criteria were infectious conditions that would interfere with the assessment. Patients were graded according to the clinical, etiological, anatomical and pathophysiological classification. Talocrural goniometry was performed to assess the degree of mobility of the legs. The Kolmogorov-Smirnov normality test, Kruskal-Wallis test, Dunn's Multiple comparison test and analysis of variance were used for statistical analysis tests with an alpha error of 5% being considered acceptable. The increase in body mass index is correlated to the reduction in joint mobility (Kruskal-Wallis test: p-value <0.0001) and increase in clinical, etiological, anatomical and pathophysiological classification is correlated to a decrease in joint mobility and the increase in age is associated with an increase in clinical, etiological, anatomical and pathophysiological classification (Kruskal-Wallis test: p-value <0.0001). Obesity is associated with deterioration in joint mobility and worsening of chronic venous disease.

Pharyngeal airway characterization in adolescents related to facial skeletal pattern: A preliminary study

The objective of this study was to characterize the volume and the morphology of the pharyngeal airway in adolescent subjects, relating them to their facial skeletal pattern. Fifty-four subjects who had cone-beam computed tomography were divided into 3 groups-skeletal Class I, Class II, and Class III-according to their ANB angles. The volumes of the upper pharyngeal portion and nasopharynx, and the volume and morphology of the lower pharyngeal portion and its subdivisions (velopharynx, oropharynx, and hypopharynx) were assessed with software (version 11.5; Dolphin Imaging & Management Solutions, Chatsworth, Calif). The results were compared with the Kruskal-Wallis and the Dunn multiple comparison tests to identify intergroup differences. Correlations between variables assessed were tested by the Spearman correlation coefficient. Correlations between the logarithms of airway volumes and the ANB angle values were tested as continuous variables with linear regression, considering the sexes as subgroups. The minimum areas in the Class II group (112.9 ± 42.9, 126.9 ± 45.9, and 142.1 ± 83.5 mm(2)) were significantly smaller than in Class III group (186.62 ± 83.2, 234.5 ± 104.9, and 231.1 ± 111.4 mm(2)) for the lower pharyngeal portion, the velopharynx, and the oropharynx, respectively, and significantly smaller than the Class I group for the velopharynx (201.8 ± 94.7 mm(2)). The Class II group had a statistically significant different morphology than did the Class I and Class III groups in the velopharynx. There was a tendency to decreased airway volume with increased ANB angle in the lower pharyngeal portion, velopharynx, and oropharynx. In the upper pharyngeal portion, nasopharynx, and hypopharynx, there seemed to be no association between the airway volume and the skeletal pattern. The Class II subjects had smaller minimum andmean areas (lower pharyngeal portion, velopharynx, and oropharynx) than did the Class III group and significantly less uniform velopharynx morphology than did the Class I and Class III groups. A negative correlation was observed between the ANB value and airway volume in the lower pharyngeal portion and the velopharynx (both sexes) and in the oropharynx (just in male subjects).

Wall shear stress effects of different endodontic irrigation techniques and systems

ObjectivesThis study examined débridement efficacy as a result of wall shear stresses created by different irrigant delivery/agitation techniques in an inaccessible recess of a curved root canal model. MethodsA reusable, curved canal cavity containing a simulated canal fin was milled into mirrored titanium blocks. Calcium hydroxide (Ca(OH)2) paste was used as debris and loaded into the canal fin. The titanium blocks were bolted together to provide a fluid-tight seal. Sodium hypochlorite was delivered at a previously-determined flow rate of 1mL/min that produced either negligible or no irrigant extrusion pressure into the periapex for all the techniques examined. Nine irrigation delivery/agitation techniques were examined: NaviTip passive irrigation control, Max-i-Probe® side-vented needle passive irrigation, manual dynamic agitation (MDA) using non-fitting and well-fitting gutta-percha points, EndoActivator™ sonic agitation with medium and large points, VPro™ EndoSafe™ irrigation system, VPro™ StreamClean™ continuous ultrasonic irrigation and EndoVac apical negative pressure irrigation. Débridement efficacies were analysed with Kruskal–Wallis ANOVA and Dunn's multiple comparisons tests (α=0.05). ResultsEndoVac was the only technique that removed more than 99% calcium hydroxide debris from the canal fin at the predefined flow rate. This group was significantly different (p<0.05) from the other groups that exhibited incomplete Ca(OH)2 removal. ConclusionsThe ability of the EndoVac system to significantly clean more debris from a mechanically inaccessible recess of the model curved root canal may be caused by robust bubble formation during irrigant delivery, creating higher wall shear stresses by a two-phase air–liquid flow phenomenon that is well known in other industrial débridement systems.

Ocular surface assessment in soft contact lens wearers; the contribution of tear osmolarity among other tests

To determine whether tear osmolarity contributes to the assessment of the ocular surface in soft contact lens (CL) wearers. Prospective, case-control series in 44 CL wearers (28 tolerant and 16 intolerant) and 34 healthy subjects. Every patient underwent a thorough ophthalmic examination with a tear osmolarity test (TearLab System), conjunctival impression cytology and meibomian lipid sampling. Symptoms, break-up time (BUT), tear osmolarity, conjunctival expression of HLA-DR and meibomian fatty acid composition were evaluated. Tear osmolarity did not differ between controls and CL wearers (p= 0.23). Flow cytometry results expressed in antibody-binding capacity (ABC) units and percentage of positive cells revealed a significant difference between the intolerant CL wearer group and the control group (p< 0.0001). Comparisons between tolerant and intolerant CL wearers showed only a significant difference for mean fluorescence levels expressed in ABC units (p< 0.0001). The BUT was significantly shorter in intolerant and tolerant CL wearers subjects than in healthy subjects (p< 0.0001), whereas there was no significant difference in meibomian fatty acid composition (p= 0.99) between the two groups. Contact lens wear is responsible for ocular surface alterations whose patterns are very similar to those reported in early dry-eye syndrome. However, tear osmolarity was not modified in these selected CL wearers. The yield of tear osmolarity with TearLab™ in assessing ocular surface disorders in CL wearers deserves further investigation.

The long‐term effect of calcium hydroxide application on dentin fracture strength of endodontically treated teeth

The aim of this in vitro study was to evaluate the long-term effect of calcium hydroxide (CH) on the microtensile fracture strength (MTFS) of endodontically treated human teeth. A total of 105 caries-free extracted human mandibular incisors were used. The teeth were divided into seven experimental groups of 15 teeth each. The root canals of all the teeth were rotary instrumented. The teeth in the control group were immediately obturated and tested for MTFS. The rest of the teeth were vertically compacted with CH and a sterile saline solution mixture and sealed with temporary filling. The teeth were stored in a moist environment for 30, 90, 180, 270, 360, and 540days. On scheduled dates, the root canals were obturated and then the teeth were tested for MTFS with an Instron testing machine. The Kruscal-Wallis test and a post hoc Dunn's multiple comparison test was utilized. The statistical significance level was established at P<0.05. The introduction of CH into the root canals seems to decrease the MTFS of teeth statistically significantly through the 180th, 270th, 360th, and 540th days, respectively, compared with the control group. The results clearly indicated that there are statistically significant differences between group 2 and group 7 (P<0.001) and between group 3 and group 7 (P<0.05). There were no statistically significant differences between the rest of the groups (P>0.05). The results indicate that long-term CH treatments can significantly reduce the strength of the teeth, causing an increase in fracture risk.

Role of TNF-related apoptosis-inducing ligand (TRAIL) in the pathogenesis of varicocele-induced testicular dysfunction

The higher frequency of varicocele in men with infertility has drawn attention and resulted in increased research at the molecular level towards treatments. The aim of this study was to investigate the role of tumor necrosis factor (TNF)-related apoptosis-inducing ligand (TRAIL) and its receptors in varicocele-induced testicular dysfunction in an experimental rat model. The rats were divided into three groups: control, sham and varicocele. Varicoceles in rats were induced by partial ligation of the left renal vein and left testes. The rats were analyzed 13 weeks after surgery. The degree of DNA fragmentation within cells in the testis was determined using terminal deoxynucleotidyl transferase deoxyuridine triphosphate nick end labeling (TUNEL) assay. Tubule degeneration was evaluated using the Johnsen score. The expression of TRAIL and its receptors was detected by immunohistochemical and Western blotting techniques. The apoptotic index, Johnsen score and the expression of TRAIL and TRAIL receptors were examined. The data are presented as the mean±s.d. and were analyzed using computer software. The Kruskal-Wallis and Dunn's multiple comparison tests were used in the statistical analyses. The germ cell apoptotic index was increased in rats with varicoceles when compared with the sham and control groups (P=0.0031). The Johnsen score was significantly decreased in the varicocele group when compared with the sham and control groups (P<0.0001). Immunohistochemical and Western blotting analyses showed that after varicocele induction, the expression of TRAIL-R1 and TRAIL-R4 in germ cells was increased and the expression of TRAIL-R2 was decreased. There are no significant differences among the groups in terms of TRAIL and TRAIL-R3 receptor expression. The results of this study indicate that TRAIL and its receptors may have a potential role in the pathogenesis of varicocele-induced testicular dysfunction.

Survival of microorganisms on complete dentures following ultrasonic cleaning combined with immersion in peroxide‐based cleanser solution

To compare ultrasonic cleaning combined with immersion in a commercially available peroxide-based cleanser solution (Polident(®) ) with other denture cleaning methods, we examined the quantity of micro-organisms that survived on dentures before and after various cleaning methods. One hundred complete dentures belonging to 50 nursing home residents (mean age, 84.6 years) were randomly assigned to five groups according to the cleaning method employed: (A) immersion in Polident(®) solution alone, (B) brushing with water, (C) ultrasonic cleaning with water, (D) method (A) followed by method (B) and (E) ultrasonic cleaning combined with immersion in Polident(®) solution. Before and after the dentures had been cleaned, denture biofilm was collected from the mucosal surface of each lateral half of the examined dentures. The collected micro-organisms were cultured, presumptively identified by standard methods and quantified. Comparisons between the five cleaning methods were carried out using the Kruskal-Wallis test and Dunn's multiple comparisons test. The denture cleaning methods involving the use of Polident(®) solution (methods A, D and E) were significantly more effective at denture disinfection than the other methods (p < 0.05); in particular, the quantity of Candida spp. was lowest after method E (median, 0.00; significantly lower than those observed after methods A, B and C; p < 0.05). It was concluded that ultrasonic cleaning combined with immersion in a peroxide-based cleanser solution effectively reduces the quantity of micro-organisms surviving on dentures and is a suitable method for elderly individuals who find brushing their dentures difficult.

Elevation of Hypercoagulability Markers in Hemoglobin SC Disease

AbstractAbstract 3227 Background:Sickle cell anemia (SCA) patients present an elevated rate of thrombotic complications and increased biological markers of hemostatic activation. Hemoglobin SC disease (HbSC) is the second most prevalent hemoglobinopathy after SCA (homozygous HbSS), has specific clinical and pathological characteristics that may differ from SCA, and viscosity appears to be a hallmark of the disease. Studies have shown an increase in thrombotic events in HbSC patients, especially pulmonary embolism, however, not much is known about the coagulation activation in this population. We herein aimed to evaluate the hypercoagulability markers in HbSC disease and their associations with patients' clinical and laboratory characteristics. Patients and Methods:This was a cross-sectional study performed on a cohort of 41 adult HbSC (mean age of 43 years) and 58 adult HbSS patients (mean age of 36 years), all in steady-state, and 25 healthy age-matched controls. We evaluated the expression of tissue factor (TF), the physiological initiator of coagulation, and thrombin-antithrombin complex (TAT), a final marker of coagulation activation. Leukocyte TFmRNA expression was analyzed by real time quantitative PCR and TAT plasma levels were measured by ELISA. Comparisons between the two patient groups and controls were performed using Kruskal-Wallis test followed by Dunn's Multiple Comparisons test. Fisher's exact test and Mann-Whitney's U test were used to compare patients' clinical complications and laboratory characteristics. Spearman's rank test was used for correlations analysis. Results:Relative TF mRNA expression was significantly up-regulated in HbSC patients when compared to controls (2.6 vs. 1.2), however levels of TF were lower in HbSC than HbSS patients (2.6 vs. 3.3) (P<0.0001). Confirming the biological relevance of TF expression, TAT plasma levels were also higher in HbSC patients in comparison to controls (4.2 vs. 2.4), and lower in HbSC than HbSS patients (4.2 vs. 7.3); (P<0.0001). In the analyses of the HbSC cohort, TAT levels presented significant positive correlations with inflammation markers: leukocyte (r=0.5; P=0.001), monocyte (r=0.4; P=0.01), and platelet counts (r=0.5; P=0.002); and hemolysis markers: reticulocyte counts (r=0.4; P=0.01) and lactate dehydrogenase levels (r=0.6, P=0.0001). We also evaluated associations between TAT levels and clinical complications: stroke, pulmonary arterial hypertension, acute thoracic syndrome, retinopathy, osteonecrosis, leg ulcers, autosplenectomy and microalbuminuria. HbSC patients with retinopathy had significantly higher TAT levels (4.7 vs. 3.9; P=0.03) when compared with patients without this complication. TAT levels were also higher in patients with autosplenectomy (4.8 vs. 3.8; P=0.004), and in patients with osteonecrosis, although this had borderline statistical significance (4.6 vs. 3.9; P=0.06). TF expression significantly correlated with monocyte counts (r=0.6; P=0.01). Conclusions:Our results indicate that HbSC disease patients present elevated coagulation activation markers when compared to controls, although not as intense as seen in SCA. Thrombotic complications in HbSC patients have been mainly linked to the hyperviscosity present in this disease. Our data suggest that inflammation and hemolysis are also important factors contributing to hemostatic activation, which may participate in the pathophysiology of very prevalent chronic complications of HbSC disease: retinopathy and osteonecrosis. Interestingly, in our cohort, patients with autosplenectomy had higher levels of pro-coagulant markers, possibly due to a higher intravascular hemolytic rate and elevated peripheral blood counts. Although HbSC disease is considered a milder form of SCA, autopsy studies have shown that mortality by pulmonary embolism is more frequent in HbSC disease than in SCA, being the second cause of mortality in these patients (Manci et al, 2003). Studies addressing the pathophysiology of coagulation activation in HbSC disease are lacking. Low numbers of cases of HbSC disease are usually included in studies focusing mainly in SCA, and the results are very variable, probably due to the small numbers of patients. In view of the high prevalence and morbimortalilty of thrombotic complications in this population, we believe that future studies should focus on a better understanding of hypercoagulability in HbSC disease. Disclosures:No relevant conflicts of interest to declare.

Increased Expression of the Inhibitory B-Cell Molecule CD22 on CD27+IgM+ B Cells Containing ABO-Antibody-Secreting Cells: A Role in Susceptibility to B-Cell Tolerance after ABO-Incompatible Infant Heart Transplantation?

Purpose: Immune immaturity allows ABO-incompatible heart transplantation (ABOi HTx) to be performed safely in infants and generally results in B-cell tolerance to donor ABO antigens. ABO antibodies are thought to arise in a T-independent (TI) manner; TI B-cell activation has been shown to be inhibited by the interaction of the inhibitory B-cell co-receptor CD22 with sialic acids on cells and tissues, leading to B-cell tolerance. It is unknown whether CD22 plays a role in regulating B-cell responses to ABO antigens in the transplant setting. Due to the generally reduced immune response to TI-antigens in early childhood, we hypothesized an enhanced role for CD22 during infancy compared to later in life. In this study we examined CD22 expression on various B-cell subsets. In addition, we performed functional assays to determine CD22 expression after B-cell stimulation and to assess the presence of ABO antigen-specific IgM antibody-secreting cells (ASC). Methods: We analyzed human splenocytes isolated from organ donors (n=41; ages 4 days - 74 years). Flow cytometric analysis was performed to quantify the expression levels of CD22, CD27, CD38, IgM and IgG on the surface of CD19+ B cells. CD27+IgM+ B cells and CD27−IgM+ B cells were isolated from human splenocytes (n=5) by automated magnetic activated cell sorting, labelled with Cell Proliferation Dye eFluor450 and stimulated with CpG plus IL-2, IL-10 and IL-15. After one week, CD22 expression was examined by flow cytometry and the frequency of ABO antigen-specific ASC detected by ELISPOT. Results: Significant differences were observed when comparing the median fluorescence intensity (MFI) of CD22 amongst various B-cell subsets (p< 0.0001; Kruskal Wallis test). Post testing using the Dunn's multiple comparison test revealed that CD27+IgM+ B cells had higher expression (p< 0.001) than other B-cell subsets. Furthermore, the MFI of CD22 on the CD27+IgM+ B cells was inversely correlated with age (p=0.001), with infant samples having the highest level of CD22, and expression decreasing with increasing age. After culture, down-regulated CD22 expression was observed in proliferating CD27+IgM+ B cells compared to non-proliferating cells. ELISPOT analysis showed that the vast majority of ABO antigen-specific ASC were derived from the CD27+IgM+ B-cell population. Conclusion: Based on previous studies showing the inhibitory role of CD22 on B-cell activation, our findings suggest that the increased expression of CD22 on the CD27+IgM+ B-cell subset, which includes precursors of ABO antigen-specific ASC, may cause these infant B cells to be more susceptible to down-regulation of B-cell signaling leading to subsequent inactivation. CD22 may therefore play an inhibitory role in immune responses to ABO antigens in the ABOi HTx setting. Studies are underway to determine the role of CD22 in B-cell signalling and regulation during infancy.