SAGE analysis demonstrates increased expression of TOSO contributing to Fas-mediated resistance in CLL

To identify novel genes involved in the molecular pathogenesis of chronic lymphocytic leukemia (CLL) we performed a serial analysis of gene expression (SAGE) in CLL cells, and compared this with healthy B cells (nCD19(+)). We found a high level of similarity among CLL subtypes, but a comparison of CLL versus nCD19(+) libraries revealed 55 genes that were over-represented and 49 genes that were down-regulated in CLL. A gene ontology analysis revealed that TOSO, which plays a functional role upstream of Fas extrinsic apoptosis pathway, was over-expressed in CLL cells. This finding was confirmed by real-time reverse transcription-polymerase chain reaction in 78 CLL and 12 nCD19(+) cases (P <.001). We validated expression using flow cytometry and tissue microarray and demonstrated a 5.6-fold increase of TOSO protein in circulating CLL cells (P =.013) and lymph nodes (P =.006). Our SAGE results have demonstrated that TOSO is a novel overexpressed antiapoptotic gene in CLL.

Determination of P-glycoprotein, MDR-related protein 1, breast cancer resistance protein, and lung-resistance protein expression in leukemic stem cells of acute myeloid leukemia

Background: The most primitive leukemic precursor in acute myeloid leukemia (AML) is thought to be the leukemic stem cell (LSC), which retains the properties of self-renewal and high proliferative capacity and quiescence of the hematopoietic stem cell. LSC seems to be immunophenotypically distinct and more resistant to chemotherapy than the more committed blasts. Considering that the multidrug resistance (MDR) constitutive expression may be a barrier to therapy in AML, we have investigated whether various MDR transporters were differentially expressed at the protein level by different leukemic subsets. Methods: The relative expression of the drug-efflux pumps P-gp, MRP, LRP, and BCRP was evaluated by mean fluorescence index (MFI) and the Kolmogorov-Smirnov analysis (D values) in five leukemic subpopulations: CD34(+)CD38(-)CD123(+) (LSCs), CD34(+)CD38(+)CD123(-), CD34(+)CD38(+)CD123(+), CD34(+)CD38(+)CD123(-), and CD34(-) mature cells in 26 bone marrow samples of CD34(+) AML cases. Results: The comparison between the two more immature subsets (LSC versus CD34(+)CD38(-)CD123(-) cells) revealed a higher P-gp, MRP, and LRP expression in LSCs. The comparative analysis between LSCs and subsets of intermediate maturation (CD34(+)CD38(+)) demonstrated the higher BCRP expression in the LSCs. In addition, P-gp expression was also significantly higher in the LSC compared to CD34(+)CD38(+)CD123(-) subpopulation. Finally, the comparative analysis between LSC and the most mature subset (CD34(-)) revealed higher MRP and LRP and lower P-gp expression in the LSCs. Conclusions: Considering the cellular heterogeneity of AML, the higher MDR transporters expression at the most immature, self-renewable, and quiescent LSC population reinforces that MDR is one of the mechanisms responsible for treatment failure. (C) 2008 Clinical Cytometry Society.

Glucocorticoid resistance in dialysis patients may impair the kidney allograft outcome

This study examines in vitro steroid sensitivity in chronic renal failure ( CRF) patients and its influence on the allograft outcome. We determined the inhibitory effect of dexamethasone ( DEX) on concanavalin A ( Con-A)-stimulated peripheral blood mononuclear cell ( PBMC) proliferation, and glucocorticoid receptor` ( GR) number of binding sites ( B-max) and affinity ( K-d) in 28 CRF patients and 40 normal healthy controls. Based on K-d values > 95th percentile from controls, patients were divided into two groups: glucocorticoid resistant ( n = 11) and glucocorticoid sensitive ( n = 17). Patients were followed during 18 months post-transplantation observing acute rejection episodes ( ARE), chronic allograft nephropathy ( CAN), allograft failure and death. The DEX concentration that caused 50% inhibition of Con-A-stimulated PBMC proliferation ( IC50) was higher in CRF than in healthy controls ( 2.2 x 10(-5) +/- 1.0 x 10(-5) versus 8.3 x 10(-6) +/- 4.2 x 10(-6) mol/ L, P = 0.02). Values of Kd ( 12.4 +/- 1.8 versus 7.2 +/- 0.9 nM) and Bmax ( 7.7 +/- 1.1 versus 4.1 +/- 0.3 fmol/ mg protein) were higher in CRF patients ( P = 0.02 and P = 0.001, respectively). There were higher incidences of ARE ( P = 0.02) and CAN ( P = 0.002) in the glucocorticoid-resistant group. Univariate and multivariate logistic regression showed that Kd was an independent predictor of ARE ( OR 8.8, P= 0.03) aswell as of CAN ( OR 16.5, P= 0.01). In conclusion, we observed glucocorticoid resistance in a subgroup of CRF patients undergoing dialysis, which led to a higher morbidity due to ARE and CAN in an 18-month follow-up period.

Assessment of two central auditory tests in elderly patients without hearing complaints

Speech understanding disorders in the elderly may be due to peripheral or central auditory dysfunctions. Asymmetry of results in dichotic testing increases with age, and may reflect on a lack of inter-hemisphere transmission and cognitive decline. Aim: To investigate auditory processing of aged people with no hearing complaints. Study design: clinical prospective. Materials and Methods: Twenty-two voluntary individuals, aged between 55 and 75 years, were evaluated. They reported no hearing complaints and had maximal auditory thresholds of 40 dB HL until 4 KHz, 80% of minimal speech recognition scores and peripheral symmetry between the ears. We used two kinds of tests: speech in noise and dichotic alternated dissyllables (SSW). Results were compared between males and females, right and left ears and between age groups. Results: There were no significant differences between genders, in both tests. Their Left ears showed worse results, in the competitive condition of SSW. Individuals aged 65 or older had poorer performances than those aged 55 to 64. Conclusion: Central auditory tests showed worse performance with aging. The employment of a dichotic test in the auditory evaluation setting in the elderly may help in the early identification of degenerative processes, which are common among these patients.

Short-Term Effects of Hydroxyethylstarch Resuscitation on Systemic and Regional Hemodynamics and Metabolism in a Brain-Dead Canine Model

Background. Hydroxyethylstarch (HES) is a synthetic polymer of glucose that has been suggested for therapeutic use in long-term plasma expansion. The aim of this study was to test the hypothesis that the infusion of a small volume of HES may provide benefits in systemic and regional hemodynamics and metabolism in a brain-dead canine model compared with large volume crystalloid resuscitation. Methods. Fourteen mongrel dogs were subjected to a brain-death protocol by consecutive insufflations of a balloon catheter in the epidural space. One hour after induction of brain-death, the animals were randomly assigned to two groups: NS (0.9% NaCl, 33mL/kg), and HES (6% HES 450/0.7, 17mL/Kg). Systemic and regional hemodynamics were evaluated using Swan-Ganz, ultrasonic flowprobes, and arterial catheters. Serial blood samples were collected for blood gas, electrolyte, and serum chemistry analysis. Systemic, hepatic, and splanchnic O(2)-derived variables were also calculated. Results. Epidural balloon insufflations induced a significant increase in mean arterial pressure, cardiac output (MAP and CO, respectively), regional blood flow, and systemic vascular resistance. Following the hyperdynamic phase, severe hypotension with normalization of systemic and regional blood flow was observed. Fluid resuscitation induced a prompt increase in MAP, CO, and portal vein blood flow, and a significant reduction in systemic and pulmonary vascular resistance. There were no differences between groups in metabolic indices, liver function tests (LFTs), or renal function tests. HES was more effective than NS in restoring cardiac performance in the first 2h after fluid resuscitation (P < 0.05). Both tested solutions partially and temporarily restored systemic and regional oxygen delivery. Conclusion. Small volumes of 6% HES 450/0.7 improved cardiovascular performance and provided the same regional hemodynamic and metabolic benefits of large volumes of isotonic crystalloid solutions. (C) 2011 Elsevier Inc. All rights reserved.

Effects of individual housing on behavior and resistance to Ehrlich tumor growth in mice

This study analyzed in Balb/C mice the effects of individual housing on behavior, serum corticosterone and resistance to Ehrlich tumor growth. Mice (60 days old) were individually (IH) or grouped housed (G) (10-12 animals/cage) for 14-21 days. The 1st day of the housing condition was considered experimental day 1 (ED1). Results showed that on ED21, IH mice, when compared to G mice, presented no differences on corticosterone serum levels when kept undisturbed; however, an increased level of this hormone was observed in IH mice after an immobilization stress challenge. An increased time spent in the plus-maze closed arms and a decreased time in the open arms were also observed in IH mice. When compared to G animals, after inoculation with 105 Ehrlich tumor cells on ED1, IH mice presented an increase in volume of ascitic fluid and number of tumor cells. The survival time of IH mice was also shorter than that measured in G animals. Furthermore, IH mice injected with a different number of tumor cells on ED1 always presented increased Ehrlich tumor cells than G group. Interestingly. these effects were not observed when the tumor cells injection was done on ED4. These results suggest that individual-housing conditions induce an altered immune-endocrine response and, at the same time, decrease animals` resistance to Ehrlich tumor growth. It is proposed that the neural link between the behavioral and immunological changes observed after the stress of individual housing might involve the activation of the HPA axis. (C) 2008 Elsevier Inc. All rights reserved.

Cyclic Fatigue Resistance of Rotary Nickel-Titanium Instruments Submitted to Nitrogen Ion Implantation

Introduction: The aim of this study was to assess cyclic fatigue resistance in rotary nickel-titanium instruments submitted to nitrogen ion implantation by using a custom-made cyclic fatigue testing apparatus. Methods: Thirty K3 files, size #25, taper 0.04, were divided into 3 experimental groups as follows: group A, 12 files exposed to nitrogen ion implantation at a dose of 2.5 x 10(17) ions/cm(2), accelerating voltage of 200 kV, currents of 1 mu A/cm(2), 130 degrees C temperature, and vacuum conditions of 10 x 10(-6) torr for 6 hours; group B, 12 nonimplanted files; and group C, 6 files submitted to thermal annealing for 6 hours at 130 degrees C. One extra file was used for process control. All files were submitted to a cyclic fatigue test that was performed with an apparatus that allowed the instruments to rotate freely, simulating rotary instrumentation of a curved canal (40-degree, 5-mm radius curve). An electric motor handpiece was used with a contra-angle of 16:1 at an operating speed of 300 rpm and a torque of 2 N-cm. Time to failure was recorded with a stopwatch in seconds and subsequently converted to number of cycles to fracture. Data were analyzed with the Student t test (P < .05). Results: Ion-implanted instruments reached significantly higher cycle numbers before fracture (mean, 510 cycles) when compared with annealed (mean, 428 cycles) and nonimplanted files (mean, 381 cycles). Conclusions: Our results showed that nitrogen ion implantation improves cyclic fatigue resistance in rotary nickel-titanium instruments. Industrial implementation. of this surface modification technique would produce rotary nickel-titanium instruments with a longer working life. (J Endod 2010;36:1183-1186)

In vitro evaluation of erbium, chromium:yttrium-scandium-gallium-garnet laser-treated enamel demineralization

This study evaluated the effect of different parameters of erbium, chromium:yttrium-scandium-gallium-garnet (Er,Cr:YSGG) laser irradiation on enamel mineral loss in a simulated caries model. Forty-five enamel samples obtained from third molar teeth (3 mmx 3 mm) were randomly divided into five groups (n = 9): G1-Er,Cr:YSGG laser at 0.25 W, 20 Hz, 2.8 J/cm(2); G2-Er,Cr:YSGG laser at 0.50 W, 20 Hz, 5.7 J/cm(2); G3-Er,Cr:YSGG laser at 0.75 W, 20 Hz, 8.5 J/cm(2); G4-sodium fluoride (NaF) dentifrice (positive control); G5-no treatment (negative control). After irradiation, the samples were submitted to 2 weeks of pH cycling. After the acid challenge, the samples were assessed by cross-sectional microhardness at different depths from the enamel surface. Analysis of variance (ANOVA) and Student-Newman-Keuls tests were performed (alpha = 5%). The percentage of lesion inhibition for each group was: G1 37%; G2 38%; G3 64%, and G4 50.5%. Regarding the relative mineral loss values (micrometers x volume percent), groups G1 (1,392 +/- 522) and G2 (1,292 +/- 657) did not differ significantly from each other, but both had higher values than group G3 (753 +/- 287); the groups irradiated with Er,Cr:YSGG laser did not differ from group G4. Although the findings of the study revealed that Er,Cr:YSGG laser irradiation at 8.5 J/cm(2) can be an alternative for the enhancement of the enamel`s resistance to acid, lower energy densities also produced a cariostatic potential comparable to the use of fluoride dentifrice.

Caries Resistance of Lased Human Root Surface with 10.6 mu m CO(2) Laser-Thermal, Morphological, and Microhardness Analysis

Although the cariostatic effects of CO(2) laser on enamel have been shown, its effects on root surface demineralization remains uncertain. The objectives of this in vitro research was to establish safe parameters for a pulsed 10.6 mu m CO(2) laser and to evaluate its effect on morphological features of the root surface, as well as on the reduction of root demineralization. Ninety-five human root surfaces were randomly divided into five groups: G1-No treatment (control); G2-2.5 J/cm(2); G3-4.0 J/cm(2); G4-5.0 J/cm(2); and G5-6.0 J/cm(2). Intrapulpal temperature was evaluated during root surface irradiation by a thermocouple and morphological changes were evaluated by SEM. After the surface treatment, the specimens were submitted to a 7-day pH-cycling model. Subsequently, the cross-sectional Knoop microhardness values were measured. For all irradiated groups, intrapulpal temperature changes were less than 1.5 degrees C. Scanning electron microscopy images indicated that fluences as low as 4.0 J/cm(2) were sufficient to induce morphological changes in the root surface. Additionally, for fluences reaching or exceeding 4.0 J/cm(2), laser-induced inhibitory effects on root surface demineralization were observed. It was concluded that laser energy density in the range of 4.0 to 6.0 J/cm(2) could be applied to a dental root to reduce demineralization of this surface without compromising pulp vitality.

Physical-mechanical properties of glass ionomer cements indicated for atraumatic restorative treatment

Background: This study evaluated mechanical properties of glass ionomer cements (GICs) used for atraumatic restorative treatment. Wear resistance, Knoop hardness (Kh), flexural (F(s)) and compressive strength (C(s)) were evaluated. The GICs used were Riva Self Cure (RVA), Fuji IX (FIX), Hi Dense (HD), Vitro Molar (VM), Maxxion R (MXR) and Ketac Molar Easymix (KME). Methods: Wear was evaluated after 1, 4, 63 and 365 days. Two-way ANOVA and Tukey post hoc tests (P = 0.05) analysed differences in wear of the GICs and the time effect. F(s), C(s), and Kh were analysed with one-way ANOVA. Results: The type of cement (p < 0.001) and the time (p < 0.001) had a significant effect on wear. In early-term wear and Kh, KME and FIX presented the best performance. In long-term wear, F(s) and C(s), KME, FIX and HD had the best performance. Strong explanatory power between F(s) and the Kh (r(2) = 0.85), C(s) and the Kh (r(2) = 0.82), long-term wear and F(s) of 24 h (r(2) = 0.79) were observed. Conclusions: The data suggested that KME and FIX presented the best in vitro performance. HD showed good results except for early-term wear.

Fluoride uptake and acid resistance of enamel irradiated with Er : YAG laser

This study evaluated the resistance to demineralization and fluoride incorporation of enamel irradiated with Er:YAG. A total of 110 bovine teeth were selected and divided into eight groups: unlased, 37% phosphoric acid, and samples irradiated with the Er:YAG laser at several fluences (31.84 J/cm(2), 25.47 J/cm(2), 19.10 J/cm(2), 2.08 J/cm(2), 1.8 J/cm(2), and 0.9 J/cm(2)). The application of acidulated phosphate fluoride was performed after treatments. All samples were immersed in 2 ml of 2.0 M acetic-acetate acid solution at pH 4.5 for 8 h, and fluoride, calcium, and phosphorus ions dissolved were analyzed by atomic absorption spectrometry and spectrophotometry. The phosphoric acid and 31.84 J/cm(2) groups presented the lowest dissolution of calcium and phosphorus ions. Higher fluoride incorporation was observed on 1.8 J/cm(2) and 0.9 J/cm(2) groups. Based on these results, Er:YAG laser was able to decrease acid dissolution and increase fluoride uptake and can be a promissory alternative for preventive dentistry.

Understanding Contradictory Data in Contraction Stress Tests

The literature shows contradictory results regarding the role of composite shrinkage and elastic modulus as determinants of polymerization stress. The present study aimed at a better understanding of the test mechanics that could explain such divergences among studies. The hypothesis was that the effects of composite shrinkage and elastic modulus on stress depend upon the compliance of the testing system. A commonly used test apparatus was simulated by finite element analysis, with different compliance levels defined by the bonding substrate (steel, glass, composite, or acrylic). Composites with moduli between 1 and 12 GPa and shrinkage values between 0.5% and 6% were modeled. Shrinkage was simulated by thermal analogy. The hypothesis was confirmed. When shrinkage and modulus increased simultaneously, stress increased regardless of the substrate. However, if shrinkage and modulus were inversely related, their magnitudes and interaction with rod material determined the stress response.

Measuring Bond Strength Between Fiber Post and Root Dentin: A Comparison of Different Tests

Purpose The aim of this study was to evaluate the ability of bond strength tests to accurately measure the bond strength of fiber posts luted into root canals Materials and Methods The test methods studied were hourglass microtensile (HM), push-out (PS), modified push out (MP) and pull out (PL) The evaluated parameters were bond strength values, reliability (using Weibull analysis), failure mode (using confocal microscopy), and stress distribution (using finite element analysis) Forty human intact single rooted and endodontically treated teeth were divided into four groups Each group was assigned one of the test methods The samples in the HM and PS groups were 1 0 +/- 0 1 mm thick, the HM samples were hourglass shaped and the PS samples were disk shaped For the PL and MP groups, each 1 mm dentin slice was luted with a fiber post piece Three dimensional models of each group were made and stress was analyzed based on Von Mises criteria Results PL provided the highest values of bond strength followed by MP both of which also had greater amounts of adhesive failures PS showed the highest frequency of cohesive failures MP showed a more homogeneous stress distribution and a higher Weibull modulus Conclusion The specimen design directly influences the biomechanical behavior of bond strength tests

Long-term effect of carbodiimide on dentin matrix and resin-dentin bonds

Objectives: To characterize the interaction of 1-Ethyl-3-[3-dimethylaminopropyl] carbodiimide Hydrochloride (EDC) with dentin matrix and its effect on the resin-dentin bond. Methods: Changes to the stiffness of demineralized dentin fragments treated with EDC/N-hydroxysuccinimide (NHS) in different solutions were evaluated at different time points. The resistance against enzymatic degradation was indirectly evaluated by ultimate tensile strength (UTS) test of demineralized dentin treated or not with EDC/NHS and subjected to collagenase digestion. Short- and long-term evaluations of the strength of resin-dentin interfaces treated with EDC/NHS for 1 h were performed using microtensile bond strength (mu TBS) test. All data (MPa) were individually analyzed using ANOVA and Tukey HSD tests (alpha = 0.05). Results: The different exposure times significantly increased the stiffness of dentin (p < 0.0001, control-5.15 and EDC/NHS-29.50), while no differences were observed among the different solutions of EDC/NHS (p = 0.063). Collagenase challenge did not affect the UTS values of EDC/NHS group (6.08) (p > 0.05), while complete degradation was observed for the control group (p = 0.0008, control-20.84 and EDC/NHS-43.15). EDC/NHS treatment did not significantly increase resin-dentin mu TBS, but the values remained stable after 12 months water storage (p < 0.05). Conclusions: Biomimetic use of EDC/NHS to induce exogenous collagen cross-links resulted in increased mechanical properties and stability of dentin matrix and dentin-resin interfaces. (C) 2010 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater 94B: 250-255, 2010.