NEF-Induced HIV-Associated Nephropathy Through HCK/LYN Tyrosine Kinases.

HIV-1-associated nephropathy (HIVAN) is a severe complication of HIV-1 infection. To gain insight into the pathogenesis of kidney disease in the setting of HIV, a transgenic (Tg) mouse model [CD4C/HIV-negative regulator factor (Nef)] was used in which HIV-1 nef expression is under control of regulatory sequences (CD4C) of the human CD4 gene, thus allowing expression in target cells of the virus. These Tg mice develop a collapsing focal segmental glomerulosclerosis associated with microcystic dilatation, similar to human HIVAN. To identify kidney cells permissive to the CD4C promoter, CD4C reporter Tg lines were used. They showed preferential expression in glomeruli, mainly in mesangial cells. Breeding CD4C/HIV Tg mice on 10 different mouse backgrounds showed that HIVAN was modulated by host genetic factors. Studies of gene-deficient Tg mice revealed that the presence of B and T cells and that of several genes was dispensable for the development of HIVAN: those involved in apoptosis (Trp53, Tnfsf10, Tnf, Tnfrsf1b, and Bax), in immune cell recruitment (Ccl3, Ccl2, Ccr2, Ccr5, and Cx3cr1), in nitric oxide (NO) formation (Nos3 and Nos2), or in cell signaling (Fyn, Lck, and Hck/Fgr). However, deletion of Src partially and that of Hck/Lyn largely abrogated its development. These data suggest that Nef expression in mesangial cells through hematopoietic cell kinase (Hck)/Lck/Yes novel tyrosine kinase (Lyn) represents important cellular and molecular events for the development of HIVAN in these Tg mice.

Experimental Investigation of Dental Composites Degradation After Early Water Exposure.

While dental composite long-term aging has already been studied in the past, no data exist about the early aging while it might be detrimental regarding the composites' longevity. This study aims to better understand the effects of early water exposure on dental composites. Dental resin composites with different fillers ratio were subjected to water exposure during 24 h, 1 week, or 1 month. After photopolymerization, the samples were stored at different conditions, whether in wet or dry condition (W, D, respectively) and in wet conditions after a first 24 h storage in dry conditions (DW). Three-point bending tests were performed to measure the flexural modulus. The samples were then subjected to a sorption/desorption protocol. While the matrix alone did not undergo any mechanical degradation with exposure time, the composites matrices presented a decrease in elastic modulus. This decrease was the highest for the matrix with nonsilanized fillers. Interestingly, the DW condition was detrimental for all the samples. Regarding the sample with nonsilanized fillers in DW for 1 month presented an elastic modulus lower than the matrix alone. These results were assigned to the sorption capacity of the polymer matrix, suggesting that the diffusion mechanisms and the nature of water molecules are determinant in the composite degradation. This study showed that dental composite early degradation mechanisms after water exposure are involved in the polymer matrix postpolymerization process as soon as after 24 h. Such mechanisms are detrimental in terms of the dental composite efficiency and have to be understood.

HIV-1 Nef Induces Hck/Lyn-Dependent Expansion of Myeloid-Derived Suppressor Cells Associated with Elevated Interleukin-17/G-CSF Levels.

Human immunodeficiency virus (HIV) or simian immunodeficiency virus (SIV) infection causes myelodysplasia, anemia, and accumulation of inflammatory monocytes (CD14+ CD16+) through largely unknown cellular and molecular pathways. The mouse cells thought to be equivalent to human CD14+ CD16+ cells are CD11b+ Gr1+ myeloid-derived suppressor cells (MDSC). We used HIV transgenic (Tg) mouse models to study MDSC, namely, CD4C/Nef Tg mice expressing nef in dendritic cells (DC), pDC, CD4+ T, and other mature and immature myeloid cells and CD11c/Nef Tg mice with a more restricted expression, mainly in DC and pDC. Both Tg strains showed expansion of granulocytic and CD11b+ Gr1low/int cells with MDSC characteristics. Fetal liver cell transplantation revealed that this expansion was stroma-independent and abrogated in mixed Tg/non-Tg 50% chimera. Tg bone marrow (BM) erythroid progenitors were decreased and myeloid precursors increased, suggesting an aberrant differentiation likely driving CD11b+ Gr1+ cell expansion, apparently cell autonomously in CD4C/Nef Tg mice and likely through a bystander effect in CD11c/Nef Tg mice. Hck was activated in Tg spleen, and Nef-mediated CD11b+ Gr1+ cell expansion was abrogated in Hck/Lyn-deficient Nef Tg mice, indicating a requirement of Hck/Lyn for this Nef function. IL-17 and granulocyte colony-stimulating factor (G-CSF) were elevated in Nef Tg mice. Increased G-CSF levels were normalized in Tg mice treated with anti-IL-17 antibodies. Therefore, Nef expression in myeloid precursors causes severe BM failure, apparently cell autonomously. More cell-restricted expression of Nef in DC and pDC appears sufficient to induce BM differentiation impairment, granulopoiesis, and expansion of MDSC at the expense of erythroid maturation, with IL-17→G-CSF as one likely bystander contributor. IMPORTANCE HIV-1 and SIV infection often lead to myelodysplasia, anemia, and accumulation of inflammatory monocytes (CD14+ CD16+), with the latter likely involved in neuroAIDS. We found that some transgenic (Tg) mouse models of AIDS also develop accumulation of mature and immature cells of the granulocytic lineage, decreased erythroid precursors, and expansion of MDSC (equivalent to human CD14+ CD16+ cells). We identified Nef as being responsible for these phenotypes, and its expression in mouse DC appears sufficient for their development through a bystander mechanism. Nef expression in myeloid progenitors may also favor myeloid cell expansion, likely in a cell-autonomous way. Hck/Lyn is required for the Nef-mediated accumulation of myeloid cells. Finally, we identified G-CSF under the control of IL-17 as one bystander mediator of MDSC expansion. Our findings provide a framework to determine whether the Nef>Hck/Lyn>IL-17>G-CSF pathway is involved in human AIDS and whether it represents a valid therapeutic target.

Children need privacy too: Respecting confidentiality in paediatric practice.

There is a paucity of published research and normative standards to guide health care providers in managing issues related to children's confidentiality. In this article, we examine challenges regarding the respect for confidentiality with children and youth in health care and offer guiding principles for clinical practice. We present an ethical framework and practice recommendations for: (a) respecting confidentiality for children who have decision-making capacity (as legally defined in different jurisdictions) and (b) recognizing some degree of respect for confidentiality among children who do not have decision-making capacity. This article will help incite additional analyses and research that can further refine the normative and practice standards proposed here.

Shear bond, wettability and AFM evaluations on CO2 laser-irradiated CAD/CAM ceramic surfaces.

The purpose of this study is to determine the CO2 laser irradiation in comparison with sandblasting (Sb), hydrofluoric acid (Hf) and silane coupling agent (Si) on shear bond strength (SBS), roughness (Rg) and wettability (Wt) of resin cement to CAD/CAM ceramics. Sixty (CAD/CAM) ceramic discs were prepared and distributed into six different groups: group A, control lithium disilicate (Li); group B, control zirconia (Zr); group C, Li: CO2/HF/Si; group D, Li: HF/Si; group E, Zr: CO2/Sb/Si; group F, Zr: Sb/Si. Result showed significant difference between irradiated and non-irradiated in terms of shear bond strength for zirconia ceramics (p value = 0.014). Moreover, partial surface wettability for irradiated and non-irradiated ceramics. Irradiated surface demonstrated more rough surface in lithium disilicate than zirconia ceramics. CO2 irradiation could increase shear bond strength, surface roughness and wettability for both CAD/CAM ceramics.

Quantification of receptor tyrosine kinase transactivation through direct dimerization and surface density measurements in single cells.

Cell signaling involves dynamic changes in protein oligomerization leading to the formation of different signaling complexes and modulation of activity. Spatial intensity distribution analysis (SpIDA) is an image analysis method that can directly measure oligomerization and trafficking of endogenous proteins in single cells. Here, we show the use of SpIDA to quantify dimerization/activation and surface transport of receptor protein kinases--EGF receptor and TrkB--at early stages of their transactivation by several G protein-coupled receptors (GPCRs). Transactivation occurred on the same timescale and was directly limited by GPCR activation but independent of G-protein coupling types. Early receptor protein kinase transactivation and internalization were not interdependent for all receptor pairs tested, revealing heterogeneity between groups of GPCRs. SpIDA also detected transactivation of TrkB by dopamine receptors in intact neurons. By allowing for time and space resolved quantification of protein populations with heterogeneous oligomeric states, SpIDA provides a unique approach to undertake single cell multivariate quantification of signaling processes involving changes in protein interactions, trafficking, and activity.

Inhibition of the Nef regulatory protein of HIV-1 by a single-domain antibody.

The Nef protein of HIV-1 is important for AIDS pathogenesis, but it is not targeted by current antiviral strategies. Here, we describe a single-domain antibody (sdAb) that binds to HIV-1 Nef with a high affinity (K(d) = 2 × 10(-9)M) and inhibited critical biologic activities of Nef both in vitro and in vivo. First, it interfered with the CD4 down-regulation activity of a broad panel of nef alleles through inhibition of the Nef effects on CD4 internalization from the cell surface. Second, it was able to interfere with the association of Nef with the cellular p21-activated kinase 2 as well as with the resulting inhibitory effect of Nef on actin remodeling. Third, it counteracted the Nef-dependent enhancement of virion infectivity and inhibited the positive effect of Nef on virus replication in peripheral blood mononuclear cells. Fourth, anti-Nef sdAb rescued Nef-mediated thymic CD4(+) T-cell maturation defects and peripheral CD4(+) T-cell activation in the CD4C/HIV-1(Nef) transgenic mouse model. Because all these Nef functions have been implicated in Nef effects on pathogenesis, this anti-Nef sdAb may represent an efficient tool to elucidate the molecular functions of Nef in the virus life cycle and could now help to develop new strategies for the control of AIDS.

Cdx2 regulation of posterior development through non-Hox targets.

The homeodomain transcription factors Cdx1, Cdx2 and Cdx4 play essential roles in anteroposterior vertebral patterning through regulation of Hox gene expression. Cdx2 is also expressed in the trophectoderm commencing at E3.5 and plays an essential role in implantation, thus precluding assessment of the cognate-null phenotype at later stages. Cdx2 homozygous null embryos generated by tetraploid aggregation exhibit an axial truncation indicative of a role for Cdx2 in elaborating the posterior embryo through unknown mechanisms. To better understand such roles, we developed a conditional Cdx2 floxed allele in mice and effected temporal inactivation at post-implantation stages using a tamoxifen-inducible Cre. This approach yielded embryos that were devoid of detectable Cdx2 protein and exhibited the axial truncation phenotype predicted from previous studies. This phenotype was associated with attenuated expression of genes encoding several key players in axial elongation, including Fgf8, T, Wnt3a and Cyp26a1, and we present data suggesting that T, Wnt3a and Cyp26a1 are direct Cdx2 targets. We propose a model wherein Cdx2 functions as an integrator of caudalizing information by coordinating axial elongation and somite patterning through Hox-independent and -dependent pathways, respectively.

HIV-1 Nef disrupts maturation of CD4+ T cells through CD4/Lck modulation.

The HIV-1 Nef protein is a major determinant of HIV-1 pathogenicity. It has been found to induce thymocyte depletion, but the mechanisms involved are not completely understood. Also, nothing is known about its effects on thymocyte selection. We used the CD4C/HIV(Nef) transgenic (Tg) mice, which develop a profound CD4(+) T cell lymphopenia, to study their thymic development. We report that HIV-1 Nef causes depletion of double-positive thymocytes and impairs selection and lineage commitment of CD4(+) single-positive thymocytes. This latter defect could be relieved by increasing the affinity of the TCR-MHC interaction or by allowing CD4(+) T cell maturation to proceed in absence of the CD4 tail, in double-Tg (Nef × CD4(tailless)) mice or in the presence of constitutively active Tg Lck(Y505F). These rescue strategies also resulted in reversal of peripheral CD4(+) T cell lymphopenia. Our data indicate that impairment of Lck-mediated CD4 coreceptor signaling by Nef is an important in vivo mechanism of HIV-1 pathogenesis.

NuA4-dependent acetylation of nucleosomal histones H4 and H2A directly stimulates incorporation of H2A.Z by the SWR1 complex.

Structural and functional analyses of nucleosomes containing histone variant H2A.Z have drawn a lot of interest over the past few years. Important work in budding yeast has shown that H2A.Z (Htz1)-containing nucleosomes are specifically located on the promoter regions of genes, creating a specific chromatin structure that is poised for disassembly during transcription activation. The SWR1 complex is responsible for incorporation of Htz1 into nucleosomes through ATP-dependent exchange of canonical H2A-H2B dimers for Htz1-H2B dimers. Interestingly, the yeast SWR1 complex is functionally linked to the NuA4 acetyltransferase complex in vivo. NuA4 and SWR1 are physically associated in higher eukaryotes as they are homologous to the TIP60/p400 complex, which encompasses both histone acetyltransferase (Tip60) and histone exchange (p400/Domino) activities. Here we present work investigating the impact of NuA4-dependent acetylation on SWR1-driven incorporation of H2A.Z into chromatin. Using in vitro histone exchange assays with native chromatin, we demonstrate that prior chromatin acetylation by NuA4 greatly stimulates the exchange of H2A for H2A.Z. Interestingly, we find that acetylation of H2A or H4 N-terminal tails by NuA4 can independently stimulate SWR1 activity. Accordingly, we demonstrate that mutations of H4 or H2A N-terminal lysine residues have similar effects on H2A.Z incorporation in vivo, and cells carrying mutations in both tails are nonviable. Finally, depletion experiments indicate that the bromodomain-containing protein Bdf1 is important for NuA4-dependent stimulation of SWR1. These results provide important mechanistic insight into the functional cross-talk between chromatin acetylation and ATP-dependent exchange of histone H2A variants.

A numerical, theoretical and experimental study of the effect of thermocycling on the matrix-filler interface of dental restorative materials.

OBJECTIVE: Thermocycling is widely used to age dental composites but with very different results from one study to another, even with apparent similar conditions. This study aims at understanding better the relative damaging speed of matrix and fillers, based on theoretical models. METHODS: Eight formulations of an experimental dental material were produced. The same organic matrix was used and silanated barium glass particles were added as fillers with different filler ratios. Samples were thermocycled up to 10 000 cycles. Three-point bending tests were carried out at different steps. The yield stress was measured among other mechanical properties. RESULTS: Composite properties were degraded by thermocycling. The decrease was slight during the first 5 000 cycles whereas it decreased significantly after 10 000 cycles. The Turcsányi model asserts that the interface yield stress is slightly affected in the first 5 000 cycles and then falls down, while the decrease of matrix yield stress is linear. SIGNIFICANCE: Each component of a composite does not age at the same rate. First, the matrix acts as a protector until the water finds its way to the interphase. The filler silanization treatment is highly sensitive to hydrolysis and is damaged rapidly from that moment. Numerical simulations and surface observations confirmed that cracks appear to propagate in the neighbourhood of the interface but not directly within it.

Nd:YAP laser in the treatment of dentinal hypersensitivity: An ex vivo study.

PURPOSE: The aims of this ex vivo study were to evaluate the effectiveness of the Nd:YAP laser in the treatment of dentin hypersensitivity, to compare the temperature rise during laser irradiation at three different dentine thicknesses, and to analyse the composition of the dentine-lased surface. METHODS: A total of 33 teeth were used in this study. For scanning electron microscopy (SEM) observation, 24 teeth were transversely sectioned and divided into 4 groups: group A was irrigated with EDTA; group B was irradiated by Nd:YAP laser with 180 mJ energy/per pulse, 0.9 W average power, and 5 Hz frequency (power density [PD] = 229 W/cm2); group C was irradiated by Nd:YAP laser with 280 mJ energy/pulse, 1.4 W average power, and 5 Hz frequency (PD = 356 W/cm2); and group D was irradiated by Nd:YAP with 360 mJ energy/pulse, 1.8 W average power, and 5 Hz frequency (PD = 458 W/cm2). Energy-dispersive spectroscopy (EDS) analysis was performed on the same teeth evaluated for SEM observations. For temperature increase evaluation performed with thermocouples, 9 teeth were transversely sectioned at 3 different thicknesses (3 for each group) of 1, 2, and 3 mm. RESULTS: Statistical analysis showed significant changes in the diameter of the dentinal tubule orifices among all groups; EDS did not show modification of the Ca/P ratio. Temperature increase under irradiation exceeded 5.5 °C only in the group D samples. CONCLUSIONS: This ex vivo study, based on temperature recording, SEM observation, and EDS analysis, demonstrated that Nd:YAP laser at a PD of 356 W/cm2, corresponding to an average power of 1.4 W, defines the best treatment for dentine hypersensitivity in terms of compromise between efficacy of the treatment and safety of the pulp.

Theoretical prediction of dental composites yield stress and flexural modulus based on filler volume ratio.

OBJECTIVE: A costly advantageous approach in composites development process is to limit experimental tests by predicting mechanical properties with respect to their filler ratio. Models exist for other fields than dentistry. They have been compared to 3-point bending test experimental results for yield stress, flexural modulus and flexural strength. METHODS: Five formulations of the same experimental material were made. They were composed of an organic matrix and different ratios of silanated barium glass particles. The samples were stored in distilled water for 24h at 37°C prior to the 3-point bending test. The Turcsányi model for yield stress was notably investigated, and SEM was used to complete data analysis. RESULTS: The yield stress showed reproducible results and a good fit with Turcsányi model with respect to filler ratio. The flexural modulus data are not scattered but did not fit with the existing models. No trend could emerge for flexural strength and strain because of scattering; these properties are more unpredictable. The SEM observations of fracture areas confirm a good matrix-filler interface quality. SIGNIFICANCE: SEM pictures validated the numerical parameter obtained from Turcsányi model. The latter therefore seems to be applicable to dental composites. Firstly, it enables to predict the evolution of the material yield stress without testing all filler ratios. Secondly, this model provides a good way to get micro-information on the matrix-filler interface from macroscopic tests. The discrepancy between flexural modulus results and theory highlighted the necessity to include an "interface quality" parameter in accurate predictive models.

Finishing and polishing effects of multiblade burs on the surface texture of 5 resin composites: microhardness and roughness testing.

OBJECTIVES: The aim of this in vitro study was to test the effect of 2 finishing-polishing sequences (QB, combining a 12/15-fluted finishing bur and an EVO-Light polisher; QWB, adding a 30-fluted polishing bur after the 12/15-fluted finishing bur used in the QB sequence) on 5 nanotech-based resin composites (Filtek Z500, Ceram X Mono, Ceram X Duo, Tetric Evoceram, and Tetric Evoceram Bulk Fill) by comparing their final surface roughness and hardness values to those of a Mylar strip control group (MS). MATERIALS AND METHODS: Twelve specimens of each nanocomposite were prepared in Teflon moulds. The surface of each resin composite was finished with QB (5 samples), QWB (5 samples), or MS (2 samples), and then evaluated (60 samples). Roughness was analysed with an optical profilometer, microhardness was tested with a Vickers indenter, and the surfaces were examined by optical and scanning electron microscopy. Data were analysed using the Kruskal-Wallis test (p < 0.05) followed by the Dunn test. RESULTS: For the hardness and roughness of nanocomposite resin, the QWB sequence was significantly more effective than QB (p < 0.05). The Filtek Z500 showed significantly harder surfaces regardless of the finishing-polishing sequence (p < 0.05). CONCLUSIONS: QWB yielded the best values of surface roughness and hardness. The hardness and roughness of the 5 nanocomposites presented less significant differences when QWB was used.

CO2 laser dentin surface treatment most effectively increased ceramic shear bond strength.

BACKGROUND AND AIMS: The purpose of this study is to determine the effect of CO2 laser irradiation in comparison with sandblasting (Sb), hydrofluoric acid (Hf) and silane coupling agent (Si) on shear bond strength (SBS) of resin cement to CAD/CAM ceramics bonded to dentin. MATERIAL AND METHODS: Forty-eight (CAD/CAM) ceramic discs were prepared and grouped by material, i.e. lithium disilicate (Emax CAD) and zirconia ceramic (Emax ZirCAD), distributed into four different groups: group A, lithium disilicate (Li) CO2/HF/Si; group B, Li: HF/Si; group C, zirconia (Zr) CO2/Sb/Si; group D, Zr: Sb/Si. RESULTS: It was shown significant difference between irradiated and non-irradiated groups in terms of shear bond strength for zirconia ceramics (p value = 0.039). CONCLUSION: CO2 laser irradiation increases shear bond strength for both CAD/CAM ceramics bonded to dentin.

Disilicate Dental Ceramic Surface Preparation by 1070 nm Fiber Laser: Thermal and Ultrastructural Analysis.

Lithium disilicate dental ceramic bonding, realized by using different resins, is strictly dependent on micro-mechanical retention and chemical adhesion. The aim of this in vitro study was to investigate the capability of a 1070 nm fiber laser for their surface treatment. Samples were irradiated by a pulsed fiber laser at 1070 nm with different parameters (peak power of 5, 7.5 and 10 kW, repetition rate (RR) 20 kHz, speed of 10 and 50 mm/s, and total energy density from 1.3 to 27 kW/cm²) and the thermal elevation during the experiment was recorded by a fiber Bragg grating (FBG) temperature sensor. Subsequently, the surface modifications were analyzed by optical microscope, scanning electron microscope (SEM), and energy dispersive X-ray spectroscopy (EDS). With a peak power of 5 kW, RR of 20 kHz, and speed of 50 mm/s, the microscopic observation of the irradiated surface showed increased roughness with small areas of melting and carbonization. EDS analysis revealed that, with these parameters, there are no evident differences between laser-processed samples and controls. Thermal elevation during laser irradiation ranged between 5 °C and 9 °C. A 1070 nm fiber laser can be considered as a good device to increase the adhesion of lithium disilicate ceramics when optimum parameters are considered.

Microhardness evaluations of CAD/CAM ceramics irradiated with CO2 or Nd:YAP laser.

BACKGROUND AND AIMS: The aim of this study was to measure the microhardness values of irradiated computer-aided design/computer-aided manufacturing (CAD/CAM) ceramics surfaces before and after thermal treatment. MATERIALS AND METHODS: Sixty CAD/CAM ceramic discs were prepared and grouped by material, i.e. lithium disilicate ceramic (Emax CAD) and zirconia ceramic (Emax ZirCAD). Laser irradiation at the material surface was performed with a carbon dioxide laser at 5 Watt (W) or 10 W power in continuous mode (CW mode), or with a neodymium:yttrium aluminum perovskite (Nd:YAP) laser at 10 W on graphite and non-graphite surfaces. Vickers hardness was tested at 0.3 kgf for lithium disilicate and 1 kgf for zirconia. RESULTS: Emax CAD irradiated with CO2 at 5 W increased microhardness by 6.32 GPa whereas Emax ZirCAD irradiated with Nd:YAP decreased microhardness by 17.46 GPa. CONCLUSION: CO2 laser effectively increases the microhardness of lithium disilicate ceramics (Emax CAD).

The effect of CO2 and Nd:YAP lasers on CAD/CAM Ceramics: SEM, EDS and thermal studies.

BACKGROUND AND AIMS: The objective of this study was to investigate the interaction of infrared laser light on Computer Aided Design and Computer Aided Manufacturing (CAD/CAM) ceramic surfaces. MATERIAL AND METHODS: Sixty CAD/CAM ceramic discs were prepared and divided into two different groups: lithiumdisilicate ceramic (IPSe.maxCADs) and Zirconia ceramic (IPSe.maxZirCADs). The laser irradiation was performed on graphite and non-graphite surfaces with a Carbon Dioxide laser at 5W and 10W power in continuous mode (CW mode) and with Neodymium Yttrium Aluminum Perovskite (Nd:YAP) laser at 10W. Surface textures and compositions were examined using Scanning Electron Microscopy (SEM), and Energy Dispersive Spectroscopy (EDS). Thermal elevation was measured by thermocouple during laser irradiation. RESULTS: The SEM observation showed a rough surface plus cracks and fissures on CO2 10W samples and melting areas in Nd:YAP samples; moreover, with CO2 5W smooth and shallow surfaces were observed. EDS analysis revealed that laser irradiation does not result in modifications of the chemical composition even if minor changes in the atomic mass percentage of the components were registered. Thermocouple showed several thermal changes during laser irradiation. CONCLUSION: CO2 and Nd:YAP lasers modify CAD/CAM ceramic surface without chemical composition modifications.

Photodynamic therapy versus ultrasonic irrigation: interaction with endodontic microbial biofilm, an ex vivo study.

INTRODUCTION: Photodynamic therapy was introduced as an adjuvant to conventional chemo-mechanical debridement during endodontic treatment to overcome the persistence of biofilms. The aim of this study was to evaluate the ability of photodynamic therapy (PDT) to disrupt an experimental microbial biofilm inside the root canal in a clinically applicable working time. MATERIALS AND METHODS: Thirty extracted teeth were prepared and then divided in three groups. All samples were infected with an artificially formed biofilm made of Enterococcus faecalis, Streptococcus salivarius, Porphyromonas gingivalis and Prevotella intermedia bacteria. First group was treated with Aseptim Plus® photo-activated (LED) disinfection system, second group by a 650 nm Diode Laser and Toluidine blue as photosensitizer, and the third group, as control group, by ultrasonic irrigation (PUI) using EDTA 17% and NaOCl 2.6% solutions. The working time for all three groups was fixed at 3 min. Presence or absence of biofilm was assessed by aerobic and anaerobic cultures. RESULTS: There was no statistically significant difference between results obtained from groups treated by Aseptim Plus® and Diode Laser (P<0.6267). In cultures of both groups there was a maximal bacterial growth. The group that was treated by ultrasonic irrigation and NaOCl and EDTA solutions had the best results (P<0.0001): there was a statistically significant reduction of bacterial load and destruction of microbial biofilm. CONCLUSION: Under the condition of this study, Photodynamic therapy could not disrupt endodontic artificial microbial biofilm and could not inhibit bacterial growth in a clinically favorable working time.

No detectable XMRV in subjects with chronic fatigue syndrome from Quebec.

We investigated the presence of XMRV in a cohort of Quebec patients with chronic fatigue syndrome (CFS). DNA was purified from activated peripheral blood mononuclear cells (PBMCs) and PCR was used to detect XMRV gag and env in 72 patients. Anti-XMRV antibodies were searched in sera of 62 patients by Western blot analysis. Attempts to detect XMRV antigens was made, using immunofluorescence with Gag anti-p30 antiserum on activated PBMC from 50 patients. Plasma viremia was measured by RT-PCR on 9 subjects. Finally, detection of infectious virus in 113 CFS subjects was made by co-culture of PHA+IL-2 activated PBMC with human LNCaP carcinoma cells, and by infecting the same susceptible cells with plasma, using a reverse transcriptase (RT) assay as a readout in both experiments. No detection of XMRV footprints nor infectious virus was detected with any of the approaches, in any of the tested individuals.