Patients with oral squamous cell carcinoma are characterized by increased frequency of suppressive regulatory T cells in the blood and tumor microenvironment

Oral squamous cell carcinoma (OSCC) is a cancerous lesion with high incidence worldwide. The immunoregulatory events leading to OSCC persistence remain to be elucidated. Our hypothesis is that regulatory T cells (Tregs) are important to obstruct antitumor immune responses in patients with OSCC. In the present study, we investigated the frequency, phenotype, and activity of Tregs from blood and lesions of patients with OSCC. Our data showed that > 80% of CD4(+)CD25(+) T cells isolated from PBMC and tumor sites express FoxP3. Also, these cells express surface Treg markers, such as GITR, CD45RO, CD69, LAP, CTLA-4, CCR4, and IL-10. Purified CD4(+)CD25(+) T cells exhibited stronger suppressive activity inhibiting allogeneic T-cell proliferation and IFN-gamma production when compared with CD4(+)CD25(+) T cells isolated from healthy individuals. Interestingly, approximately 25% of CD4(+)CD25(-) T cells of PBMC from patients also expressed FoxP3 and, although these cells weakly suppress allogeneic T cells proliferative response, they inhibited IFN-gamma and induced IL-10 and TGF-beta secretion in these co-cultures. Thus, our data show that Treg cells are present in OSCC lesions and PBMC, and these cells appear to suppress immune responses both systemically and in the tumor microenvironment.

Characterization of CD4(+)CD25(+) natural regulatory T cells in the inflammatory infiltrate of human chronic periodontitis

Periodontitis is an infectious disease, where putative periodontopathogens trigger chronic inflammatory and immune responses against periodontal structures, in which an unbalanced host response is also determinant to the disease outcome. It is reasonable to assume that patient susceptibility to periodontal tissue destruction could be determined by the balance between the response against periodontopathogens and regulatory mechanisms of these events mediated by suppressive T cells. In the present study, we identified and characterized natural regulatory T cells ( Tregs) in the inflammatory infiltrate of human chronic periodontitis ( CP) with emphasis on phenotypic analyses that were carried out to address the participation of Tregs in CP. Results showed that patients with CP presented increased frequency of T lymphocytes and CD4(+)CD25(+) T cells in the inflammatory infiltrate of gingival tissues. These cells exhibited the phenotypic markers of Tregs such as forkhead box p3 ( Foxp3), CTLA- 4, glucocorticoidinducible TNFR, CD103, and CD45RO and seemed to be attracted to the inflammation site by the chemokines CCL17 and CCL22, as their expression and its receptor CCR4 were increased in CP patients. Moreover, besides the increased detection of Foxp3 mRNA, diseased tissues presented high expression of the regulatory cytokines IL-10 and TGF-beta. In addition, the inflammatory infiltrate in CP biopsies was composed of CD25(+)Foxp3(+) and CD25(+)TGF-beta(+) cells, thus corroborating the hypothesis of the involvement of Tregs in the pathogenesis of CP. Finally, these results indicate that Tregs are found in the chronic lesions and must be involved in the modulation of local immune response in CP patients.

CCR5-dependent regulatory T cell migration mediates fungal survival and severe immunosuppression

Paracoccidioidomycosis, a debilitating pulmonary mycosis, is caused by the dimorphic fungus Paracoccidioides brasiliensis. The infection results in the formation of granulomas containing viable yeast cells that are the fungal sources for disease reactivation. Because CD4(+)CD25(+) regulatory T cells (Tregs) are in the lesions of patients with paracoccidioidomycosis, the migration of Treg cells is dependent on the axis chemokine-chemokine receptors, and CCR5 ligands are produced in P. brasiliensis-induced lesions, we investigated the role of CCR5 in the control of the infection. The results showed that CCR5(-/-) mice are more efficient in controlling fungal growth and dissemination and exhibited smaller granulomas than wild-type (WT) mice. In the absence of CCR5, the percentage of CD4(+)CD25(+) T cells expressing Foxp3, glucocorticoid-induced TNFR (GITR), CD103, CD45(low), and CTLA-4 in the granulomas was significantly decreased. Interestingly, P. brasiliensis infection resulted in an absence of T cell proliferation in response to Con A in WT but not CCR5(-/-) mice that was abrogated by anti-CTLA-4 mAb and anti-GITR mAb. Moreover, the adoptive transfer of CD4(+)CD25(+) but not CD4(+)CD25(-) T cells from infected WT to infected CCR5(-/-) mice resulted in a significant increase in fungal load. Overall, CCR5 is a key receptor for the migration of Treg cells to the site of P. brasiliensis infections leading to down-modulation of effector immune response and the long-term presence of the fungus in the granulomas. Thus, a tight control of Treg cell migration to the granulomatous lesions could be an important mechanism for avoiding exacerbation and reactivation of the disease.

The Effect of Framework Design on Fracture Resistance of Metal-Ceramic Implant-Supported Single Crowns

This study evaluated the effect of framework design on the fracture resistance of metal-ceramic implant-supported crowns. Screw-retained molar crowns with a screw access hole composed of metal or porcelain were compared to a cement-retained crown (control). For each group (n = 10), five crowns were subjected to dynamic loading (1,200,000 x 100 N x 2 Hz at 37 degrees C). Afterward, all specimens were loaded to failure using a universal testing machine. Significant differences could be established between the cement-and screw-retained groups (P <= .05), but no difference was found between the screw-retained groups and the specimens subjected to dynamic loading. Occlusal discontinuity of screw-retained crowns affects their resistance, and the metallic support on the screw access hole did not reinforce the crowns. Int J Prosthodont 2010;23:350-352.

Effect of cantilever length and framework alloy on the stress distribution of mandibular-cantilevered implant-supported prostheses

The purpose of this in vitro study was to analyze the stress distribution on components of a mandibular-cantilevered implant-supported prosthesis with frameworks cast in cobalt-chromium (Co-Cr) or palladium-silver (Pd-Ag) alloys, according to the cantilever length. Frameworks were fabricated on (Co-Cr) and (Pd-Ag) alloys and screwed into standard abutments positioned on a master-cast containing five implant replicas. Two linear strain gauges were fixed on the mesial and distal aspects of each abutment to capture deformation. A vertical static load of 100 N was applied to the cantilever arm at the distances of 10, 15, and 20 mm from the center of the distal abutment and the absolute values of specific deformation were recorded. Different patterns of abutment deformation were observed according to the framework alloy. The Co-Cr alloy framework resulted in higher levels of abutment deformation than the silver-palladium alloy framework. Abutment deformation was higher with longer cantilever extensions. Physical properties of the alloys used for framework interfere with abutment deformations patterns. Excessively long cantilever extensions must be avoided. To cite this article:Jacques LB, Moura MS, Suedam V, Souza EAC, Rubo JH. Effect of cantilever length and framework alloy on the stress distribution of mandibular-cantilevered implant-supported prostheses.Clin. Oral Impl. Res. 20, 2009; 737-741.doi: 10.1111/j.1600-0501.2009.01712.x.

Effect of abutment`s height and framework alloy on the load distribution of mandibular cantilevered implant-supported prosthesis

In cantilevered implant-supported complete prosthesis, the abutments` different heights represent different lever arms to which the abutments are subjected resulting in deformation of the components, which in turn transmit the load to the adjacent bone. The purpose of this in vitro study was to quantitatively assess the deformation of abutments of different heights in mandibular cantilevered implant-supported complete prosthesis. A circular steel master cast with five perforations containing implant replicas (O3.75 mm) was used. Two groups were formed according to the types of alloy of the framework (CoCr or PdAg). Three frameworks were made for each group to be tested with 4, 5.5 and 7 mm abutments. A 100 N load was applied at a point 15 mm distal to the center of the terminal implant. Readings of the deformations generated on the mesial and distal aspects of the abutments were obtained with the use of strain gauges. Deformation caused by tension and compression was observed in all specimens with the terminal abutment taking most of the load. An increase in deformation was observed in the terminal abutment as the height was increased. The use of an alloy of higher elastic modulus (CoCr) also caused the abutment deformation to increase. Abutment`s height and framework alloy influence the deformation of abutments of mandibular cantilevered implant-supported prosthesis. To cite this article:Suedam V, Capello SouzaEA, Moura MS, Jacques LB, Rubo JH. Effect of abutment`s height and framework alloy on the load distribution of mandibular cantilevered implant-supported prosthesis. Clin. Oral Impl. Res. 20, 2009; 196-200.doi: 10.1111/j.1600-0501.2008.01609.x.

Fatigue life and failure modes of crowns systems with a modified framework design

Objectives: To evaluate the effect of framework design on the fatigue life and failure modes of metal ceramic (MC, Ni-Cr alloy core, VMK 95 porcelain veneer), glass-infiltrated alumina (ICA, In-Ceram Alumina/VM7), and veneered yttria-stabilized tetragonal zirconia polycrystals (Y-TZP, IPSe.max ZirCAD/IPS e.max,) crowns. Methods: Sixty composite resin tooth replicas of a prepared maxillary first molar were produced to receive crowns systems of a standard (MCs, ICAs, and Y-TZPs, n = 10 each) or a modified framework design (MCm, ICAm, and Y-TZPm, n = 10 each). Fatigue loading was delivered with a spherical steel indenter (3.18 mm radius) on the center of the occlusal surface using r-ratio fatigue (30-300 N) until completion of 10(6) cycles or failure. Fatigue was interrupted every 125,000 cycles for damage evaluation. Weibull distribution fits and contour plots were used for examining differences between groups. Failure mode was evaluated by light polarized and SEM microscopy. Results: Weibull analysis showed the highest fatigue life for MC crowns regardless of framework design. No significant difference (confidence bound overlaps) was observed between ICA and Y-TZP with or without framework design modification. Y-TZPm crowns presented fatigue life in the range of MC crowns. No porcelain veneer fracture was observed in the MC groups, whereas ICAs presented bulk fracture and ICAm failed mainly through the veneer. Y-TZP crowns failed through chipping within the veneer, without core fractures. Conclusions: Framework design modification did not improve the fatigue life of the crown systems investigated. Y-TZPm crowns showed comparable fatigue life to MC groups. Failure mode varied according to crown system. (C) 2010 Elsevier Ltd. All rights reserved.

Effect of framework design on crown failure

This study evaluated the effect of core-design modification on the characteristic strength and failure modes of glass-infiltrated alumina (In-Ceram) (ICA) compared with porcelain fused to metal (PFM). Premolar crowns of a standard design (PFMs and ICAs) or with a modified framework design (PFMm and ICAm) were fabricated, cemented on dies, and loaded until failure. The crowns were loaded at 0.5 mm min(-1) using a 6.25 mm tungsten-carbide ball at the central fossa. Fracture load values were recorded and fracture analysis of representative samples were evaluated using scanning electron microscopy. Probability Weibull curves with two-sided 90% confidence limits were calculated for each group and a contour plot of the characteristic strength was obtained. Design modification showed an increase in the characteristic strength of the PFMm and ICAm groups, with PFM groups showing higher characteristic strength than ICA groups. The PFMm group showed the highest characteristic strength among all groups. Fracture modes of PFMs and of PFMm frequently reached the core interface at the lingual cusp, whereas ICA exhibited bulk fracture through the alumina core. Core-design modification significantly improved the characteristic strength for PFM and for ICA. The PFM groups demonstrated higher characteristic strength than both ICA groups combined.

A framework for valuing derivative securities

This paper develops a general framework for valuing a wide range of derivative securities. Rather than focusing on the stochastic process of the underlying security and developing an instantaneously-riskless hedge portfolio, we focus on the terminal distribution of the underlying security. This enables the derivative security to be valued as the weighted sum of a number of component pieces. The component pieces are simply the different payoffs that the security generates in different states of the world, and they are weighted by the probability of the particular state of the world occurring. A full set of derivations is provided. To illustrate its use, the valuation framework is applied to plain-vanilla call and put options, as well as a range of derivatives including caps, floors, collars, supershares, and digital options.

Identification of the regulatory subunit of Arabidopsis thaliana acetohydroxyacid synthase and reconstitution with its catalytic subunit

Acetohydroxyacid synthase (EC 4.1.3.18; AHAS) catalyzes the initial step in the formation of the branched-chain amino acids. The enzyme from most bacteria is composed of a catalytic subunit, and a smaller regulatory subunit that is required for full activity and for sensitivity to feedback regulation by valine. A similar arrangement was demonstrated recently for yeast AHAS, and a putative regulatory subunit of tobacco AHAS has also been reported. In this latter case, the enzyme reconstituted from its purified subunits remained insensitive to feedback inhibition, unlike the enzyme extracted from native plant sources. Here we have cloned, expressed in Escherichia coil, and purified the AHAS regulatory subunit of Ambidopsis thaliana. Combining the protein with the purified A. thaliana catalytic subunit results in an activity stimulation that is sensitive to inhibition by valine, leucine, and isoleucine. Moreover, there is a strong synergy between the effects of leucine and valine, which closely mimics the properties of the native enzyme. The regulatory subunit contains a sequence repeat of approximately 180 residues, and we suggest that one repeat binds leucine while the second binds valine or isoleucine. This proposal is supported by reconstitution studies of the individual repeats, which were also cloned, expressed, and purified. The structure and properties of the regulatory subunit are reminiscent of the regulatory domain of threonine deaminase (EC 4.2.1.16), and it is suggested that the two proteins are evolutionarily related.

An empirical framework for studying desistance as a process

Recent reviews of the desistance literature have advocated studying desistance as a process, yet current empirical methods continue to measure desistance as a discrete state. In this paper, we propose a framework for empirical research that recognizes desistance as a developmental process. This approach focuses on changes in the offending rare rather than on offending itself We describe a statistical model to implement this approach and provide an empirical example. We conclude with several suggestions for future research endeavors that arise from our conceptualization of desistance.

Are pioneer axons guided by regulatory gene expression domains in the zebrafish forebrain? High-resolution analysis of the patterning of the zebrafish brain during axon tract formation

Although the principles of axon growth are well understood in vitro the mechanisms guiding axons in vivo are less clear. It has been postulated that growing axons in the vertebrate brain follow borders of neuroepithelial cells expressing specific regulatory genes. In the present study we reexamined this hypothesis by analysing the earliest growing axons in the forebrain of embryonic zebrafish. Confocal laser scanning microscopy was used to determine the spatiotemporal relationship between growing axons and the expression pattern of eight regulatory genes in zebrafish brain. Pioneer axons project either longitudinally or dorsoventrally to establish a scaffold of axon tracts during this developmental period. Each of the regulatory genes was expressed in stereotypical domains and the borders of some were oriented along dorsoventral and longitudinal planes. However, none of these borders clearly defined the trajectories of pioneer axons. In two cases axons coursed in proximity to the borders of shh and pax6, but only for a relatively short portion of their pathway. Only later growing axons were closely apposed to the borders of some gene expression domains. These results suggest that pioneer axons in the embryonic forebrain do not follow continuous pathways defined by the borders of regulatory gene expression domains, (C) 2000 Academic Press.

Adult mouse intrinsic laryngeal muscles express high levels of the myogenic regulatory factor, MYF-5

The intrinsic laryngeal muscles display unique structural and functional characteristics that distinguish them from the skeletal muscle of the trunk and limbs. These features include relatively small muscle fibers, super-fast contraction speed, and fatigue resistance. The molecular basis of tissue-specific functions and other characteristics is differential gene expression. Accordingly, we have investigated the molecular basis of the functional specialization of the intrinsic laryngeal muscles by examining the expression of two key genes in the larynx, known to be important for skeletal muscle development and function: (a) the muscle regulatory factor, Myf-5, and (b) the superfast-contracting myosin heavy chain (EO-MyHC). We have found that the adult thyroarytenoid muscles express much higher levels of both Myf-5 and EO-MyHC messenger ribonucleic acid (mRNA), compared to lower hindlimb skeletal muscle where Myf-5 mRNA levels are very low and EO-MyHC is not detectable. These findings suggest that the unique functional characteristics of the intrinsic laryngeal muscles may be based in laryngeal muscle-specific gene expression directed by a unique combination of muscle regulatory factors. Such laryngeal muscle-specific genes may allow the future development of new treatments for laryngeal muscle dysfunction.