Melan-A/MART-1-specific CD4 T cells in melanoma patients: identification of new epitopes and ex vivo visualization of specific T cells by MHC class II tetramers.

Over the past decade, many efforts have been made to identify MHC class II-restricted epitopes from different tumor-associated Ags. Melan-A/MART-1(26-35) parental or Melan-A/MART-1(26-35(A27L)) analog epitopes have been widely used in melanoma immunotherapy to induce and boost CTL responses, but only one Th epitope is currently known (Melan-A51-73, DRB1*0401 restricted). In this study, we describe two novel Melan-A/MART-1-derived sequences recognized by CD4 T cells from melanoma patients. These epitopes can be mimicked by peptides Melan-A27-40 presented by HLA-DRB1*0101 and HLA-DRB1*0102 and Melan-A25-36 presented by HLA-DQB1*0602 and HLA-DRB1*0301. CD4 T cell clones specific for these epitopes recognize Melan-A/MART-1+ tumor cells and Melan-A/MART-1-transduced EBV-B cells and recognition is reduced by inhibitors of the MHC class II presentation pathway. This suggests that the epitopes are naturally processed and presented by EBV-B cells and melanoma cells. Moreover, Melan-A-specific Abs could be detected in the serum of patients with measurable CD4 T cell responses specific for Melan-A/MART-1. Interestingly, even the short Melan-A/MART-1(26-35(A27L)) peptide was recognized by CD4 T cells from HLA-DQ6+ and HLA-DR3+ melanoma patients. Using Melan-A/MART-1(25-36)/DQ6 tetramers, we could detect Ag-specific CD4 T cells directly ex vivo in circulating lymphocytes of a melanoma patient. Together, these results provide the basis for monitoring of naturally occurring and vaccine-induced Melan-A/MART-1-specific CD4 T cell responses, allowing precise and ex vivo characterization of responding T cells.

Melanosomal targeting sequences from gp100 are essential for MHC class II-restricted endogenous epitope presentation and mobilization to endosomal compartments

CD4+ T lymphocytes play an important role in CD8+ T cell-mediated responses against tumors. Considering that about 20% of melanomas express major histocompatibility complex (MHC) class II, it is plausible that concomitant antigenic presentation by MHC class I and class II complexes shapes positive (helper T cells) or negative (regulatory T cells) anti-tumor responses. Interestingly, gp100, a melanoma antigen, can be presented by both MHC class I and class II when expressed endogenously, suggesting that it can reach endosomal/MHC class II compartments (MIIC). Here, we demonstrated that the gp100 putative amino-terminal signal sequence and the last 70 residues in carboxy-terminus, are essential for MIIC localization and MHC class II presentation. Confocal microscopy analyses confirmed that gp100 was localized in LAMP-1+ endosomal/MIIC. Gp100-targeting sequences were characterized by deleting different sections in the carboxy-terminus (residues 590 to 661). Transfection in 293T cells, expressing MHC class I and class II molecules, revealed that specific deletions in carboxy-terminus resulted in decreased MHC class II presentation, without effects on MHC class I presentation, suggesting a role in MIIC trafficking for these deleted sections. Then, we used these gp100-targeting sequences to mobilize the green fluorescent protein (GFP) to endosomal compartments, and to allow MHC class II and class I presentation of minimal endogenous epitopes. Thus, we concluded that these specific sequences are MIIC targeting motifs. Consequently, these sequences could be included in expression cassettes for endogenously expressed tumor or viral antigens to promote MHC class II and class I presentation and optimize in vivo T cell responses, or as an in vitro tool for characterization of new MHC class II epitopes.

Les appareils myofonctionnels de correction de la Classe II : l’EMG.

Introduction: La correction de la Classe II avec un appareil myofonctionnel est un traitement commun chez les patients en croissance. Le Twin Block et le correcteur de Classe II fixe (CCF) sont des appareils populaires et plusieurs publications scientifiques ont décrit leurs effets sur les tissus orofaciaux. Plusieurs articles rapportent les changements de l’électromyographie des muscles de la mastication durant le traitement avec un Twin Block, mais peu d’articles ont étudié ces changements avec un CCF. Comme le Twin Block et le CCF ont des biomécaniques différentes, leur influence sur les muscles est possiblement différente. Objectifs: Évaluer les adaptations musculaires suite à un traitement par appareil myofonctionnel : Twin Block et CCF. Matériels et méthodes: Dans une étude cohorte prospective, 24 patients en pic de croissance ont été assignés aléatoirement à un traitement (13 Twin Block; 11 CCF) et l’EMG des muscles masséters et temporaux a été mesurée à 1, 5, 13, 21, 29, 37 semaines. Les muscles ont été mesurés sous trois états: au repos, en occlusion centré (OC) et en contraction volontaire maximal (CVM) Résultats: Les données ont été analysées à l’aide d’un modèle mixte linéaire à mesures répétées et ont été documentées pour chaque muscle selon quatre conditions: i- avec Twin Block en bouche, ii- sans Twin Block en bouche iii- avec CCF en bouche et iv- sans Twin Block comparé au groupe avec CCF. Dans la condition i, des résultats significatifs ont été observés au repos pour le masséter droit et gauche, ainsi que le temporal gauche avec une valeur-p≤0.005. En CVM, la condition i montre aussi des résultats significatifs pour le masséter droit et le temporal gauche avec une valeur-p≤0.05. Les conditions ii et iii ont obtenu des résultats non-significatifs en tout temps. Par contre, lorsque ces deux conditions sont comparées l’une à l’autre (condition iv), des résultats significatifs ont été obtenus en OC pour les temporaux gauche et droit avec une valeur-p=0.005. Conclusions: Avec le Twin Block en bouche, l’EMG augmente au cours du temps en CVM, mais diminue en OC. Par contre, sans le Twin Block en bouche et avec le CCF en bouche, l’EMG ne varie pas. Cependant, le Twin Block et le CCF sont différents au niveau des mesures de l’EMG au cours des neuf mois de traitement. Ceci peut être expliqué par le nivellement graduel de l’occlusion postérieure durant le traitement avec le CCF qui ne se produit pas avec le Twin Block.

Shared HLA Class II in Six Autoimmune Diseases in Latin America: A Meta-Analysis

The prevalence and genetic susceptibility of autoimmune diseases (ADs) may vary depending on latitudinal gradient and ethnicity. The aims of this study were to identify common human leukocyte antigen (HLA) class II alleles that contribute to susceptibility to six ADs in Latin Americans through a meta-analysis and to review additional clinical, immunological, and genetic characteristics of those ADs sharing HLA alleles. DRB1∗03:01 (OR: 4.04; 95%CI: 1.41–11.53) was found to be a risk factor for systemic lupus erythematosus (SLE), Sjogren’s syndrome (SS), and type 1 diabetes mellitus (T1D). DRB1 ¨ ∗04:05 (OR: 4.64; 95%CI: 2.14–10.05) influences autoimmune hepatitis (AIH), rheumatoid arthritis (RA), and T1D; DRB1∗04:01 (OR: 3.86; 95%CI: 2.32–6.42) is a susceptibility factor for RA and T1D. Opposite associations were found between multiple sclerosis (MS) and T1D. DQB1∗06:02 and DRB1∗15 alleles were risk factors for MS but protective factors for T1D. Likewise, DQB1∗06:03 allele was a risk factor for AIH but a protective one for T1D. Several common autoantibodies and clinical associations as well as additional shared genes have been reported in these ADs, which are reviewed herein. These results indicate that in Latin Americans ADs share major loci and immune characteristics.

Are class II phosphoinositide 3-kinases potential targets for anticancer therapies?

Of the three classes of true phosphoinositide (PI) 3-kinases, the class II subdivision, which consists of three isoforms, PI3K-C2alpha, PI3K-C2beta and PI3K-C2gamma, is the least well understood. There are a number of reasons for this. This class of PI 3-kinase was identified exclusively by PCR and homology cloning approaches and not on the basis of cellular function. Like class I PI 3-kinases, class II PI 3-kinases are activated by diverse receptor types. To complicate the elucidation of class II PI 3-kinase function further, their in vitro substrate specificity is intermediate between the receptor activated class I PI 3-kinases and the housekeeping class III PI 3-kinase. The class II PI 3-kinases are inhibited by the two commonly used PI 3-kinase family selective inhibitors, wortmannin and LY294002, and there are no widely available, specific inhibitors for the individual classes or isoforms. Here the current state of understanding of class II PI 3-kinase function is reviewed, followed by an appraisal as to whether there is enough evidence to suggest that pharmaceutical companies, who are currently targeting the class I PI 3-kinases in an attempt to generate anticancer agents, should also consider targeting the class II PI 3-kinases.

Class II phosphoinositide 3-kinase defines a novel signaling pathway in cell migration

The lipid products of phosphoinositide 3-kinase (PI3K) are involved in many cellular responses such as proliferation, migration, and survival. Disregulation of PI3K-activated pathways is implicated in different diseases including cancer and diabetes. Among the three classes of PI3Ks, class I is the best characterized, whereas class II has received increasing attention only recently and the precise role of these isoforms is unclear. Similarly, the role of phosphatidylinositol-3-phosphate (PtdIns-3-P) as an intracellular second messenger is only just beginning to be appreciated. Here, we show that lysophosphatidic acid (LPA) stimulates the production of PtdIns-3-P through activation of a class II PI3K (PI3K-C2β). Both PtdIns-3-P and PI3K-C2β are involved in LPA-mediated cell migration. This study is the first identification of PtdIns-3-P and PI3K-C2β as downstream effectors in LPA signaling and demonstration of an intracellular role for a class II PI3K. Defining this novel PI3K-C2β- PtdIns-3-P signaling pathway may help clarify the process of cell migration and may shed new light on PI3K-mediated intracellular events.

Nd : YAG laser clinical assisted in class II furcation treatment

The Nd:YAG laser efficacy associated with conventional treatment for bacterial reduction has been investigated throughout literature. The purpose of this study was to evaluate the bacterial reduction after Nd:YAG laser irradiation associated with scaling and root planning in class II furcation defects in patients with chronic periodontitis. Thirty-four furcation lesions were selected from 17 subjects. The control group received conventional treatment, and the experimental group received the same treatment followed by Nd:YAG laser irradiation (100 mJ/pulse; 15 Hz; 1.5 W, 60 s, 141.5 J/cm(2)). Both treatments resulted in improvements of most clinical parameters. A significant reduction of colony forming unit (CFU) of total bacteria number was observed in both groups. The highest reduction was noted in the experimental group immediately after the treatment. The number of dark pigmented bacteria and the percentage of patients with Porphyromonas gingivalis, Prevotella intermedia, and Actinobacillus actinomycetemcomitans reduced immediately after the treatment and returned to values close to the initial ones 6 weeks after the baseline for both groups. The Nd:YAG laser associated with conventional treatment promoted significant bacterial reduction in class II furcation immediately after irradiation, although this reduction was not observed 6 weeks after the baseline.

Molecular analysis of a mutant Anticarsia gemmatalis multiple nucleopolyhedrovirus (AgMNPV) shows an interruption of an inhibitor of apoptosis gene (iap-3) by a new class-II piggyBac-related insect transposon

A new piggyBac-related transposable element (TE) was found in the genome of a mutant Anticarsia gemmatalis multiple nucleopolyhedrovirus interrupting an inhibitor of apoptosis gene. This mutant virus induces apoptosis upon infection of an Anticarsia gemmatalis cell line, but not in a Trichoplusia ni cell line. The sequence of the new TE (which was named IDT for iap disruptor transposon) has 2531 bp with two DNA sequences flanking a putative Transposase (Tpase) ORF of 1719 bp coding for a protein with 572 amino acids. These structural features are similar to the piggyBac TE, also reported for the first time in the genome of a baculovirus. We have also isolated variants of this new TE from different lepidopteran insect cells and compared their Tpase sequences.

Prevalência de oclusopatias e caracteríticas cefalométricas e dentais de escolares cearense da cidade de Fortaleza

The people of Ceará state are descended from miscegenation between the Portuguese colonizers and the native population, resulting in a different facial pattern from other populations. It is important that this pattern be thoroughly understood, along with its minimum and maximum values so that they can be assessed and respected, allowing professionals who deal with the craniofacial complex to work more efficiently and scientifically. Aim: To characterize the morphological pattern of individuals from Ceará state, whose father and grandfather are also native from Ceará, in the 10-12 year age group, not submitted to previous orthodontic treatment, in order to determine: 1) the prevalence of occlusal pattern; 2) the prevalence of dental anomalies (DA) and, 3) the skeletal and dental cephalometric characteristics of individuals that present with normal occlusion and harmonious facial pattern. Methodology: A list of 10-12 year-olds was obtained from 515 schools containing 162,713 students (Education Secretariat of Ceará State), from which 234 individuals were examined (107 boys and 157 girls). The assessment criteria adopted were: 1) Angle s Classification System to determine occlusal pattern. The occlusal characteristics were measured through overbite, overjet, crowding and interincisal diastema. 2) DA are anomalies of number, shape, size, eruption and structure and, 3) in the group that presented with normal occlusion, we used cephalometric analysis measures proposed by Downs, Steiner, Tweed, Holdaway, Jacobson and McNamara. Results: 1) 25.8% of the schoolchildren had normal occlusion, 47.5% class I malocclusion, 22.3% class II malocclusion and 4.2% class III malocclusion. No statistically significant difference was found between the age group studied and sex. Thirty percent of the individuals had normal overbite, while 36.7% and 19.7% had increased and reduced overbite, respectively. Normal overjet was found in 33.7% of the individuals, increased overjet in 50% and reduced in 16.3%. Dental crowding was observed in 62.5% of the individuals and the presence of interincisal diastema in 14.8%. 2) The prevalence of DA was 56.1%, 6.8% in the number, 10.8% in shape, 4.1% in size, 34.5% in eruption, 26.4% in structure and 17.4% had more than one DA. No association was found between DA and sex, but DA was significantly associated to malocclusion (p<0.05); 3) there was no association between sex or facial type between the measures of nasal-labial angle, position and effective maxillary length, effective mandibular length and the sagittal relationship between the molars, overjet and overbite, position of upper incisors, lower incisors and between the incisors themselves. There was a difference between sex, on the VERT index and in lower anterior facial height, upper incisor inclination and line-H, between facial types for the occlusal plane angles, mandibular plane, facial axis, lower incisor inclination, mandibular position, upper incisor position, lower anterior facial height, ANB and line-H. It was concluded that: 1) the most prevalent occlusal type was class I malocclusion, with no distinction for sex or age group, and the assessment of occlusal characteristics showed that excessive overbite and overjet were the most predominant findings, along with a high occurrence of tooth crowding; 2) a high prevalence of DA was found, particularly eruption anomalies, not influenced by sex but significantly associated to malocclusion and 3) individuals from Ceará are predominantly brachyfacial, exhibiting a number of similarities inherent to their facial pattern, such as a convex profile, retracted jaw, reduced lower third and protruded lower incisors. This study was multidisciplinary, involving researchers from the areas of epidemiology, radiology and dentistry, thereby meeting the multidisciplinarity requirements of the Postgraduate Program in Health Sciences

Distalização dos molares superiores com aparelho Pendex: estudo em modelos de gesso

OBJETIVO: a presente pesquisa clínica, longitudinal e prospectiva, avaliou os efeitos do aparelho Pendex mediante o emprego de modelos de gesso. METODOLOGIA: utilizou-se uma amostra de 30 modelos de gesso, obtidos no início e no final da distalização dos molares superiores com o aparelho Pendex, de 15 jovens brasileiros tratados no Curso de Pós-Graduação, nível de Mestrado em Ortodontia, da Faculdade de Odontologia de Araçatuba - UNESP. RESULTADOS E CONCLUSÕES: os resultados mostraram aumento do perímetro do arco dentário superior com inclinação distal e controle transversal dos primeiros molares superiores.

Effect of different adhesive systems on microleakage in class II composite resin restorations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Skeletal and dental components of Class II correction with the bionator and removable headgear splint appliances

The purpose of this study was to differentiate the dentoalveolar and skeletal effects to better understand orthodontic treatment. We evaluated the treatment changes associated with the bionator and the removable headgear splint (RHS). Methods: The sample comprised 51 consecutively treated Class II patients from 1 office who had all been successfully treated with either a bionator (n = 17) or an RHS appliance (n = 17). Class II patients waiting to start treatment later served as controls (n = 17). A modified version of the Johnston pitchfork analysis was used to quantify the dentoalveolar and skeletal contributions to the anteroposterior correction at the levels of the molars and the incisors. Results: Both appliances significantly improved anteroposterior molar relationships (2.15 mm for the bionator, 2.27 mm for the RHS), primarily by dentoalveolar modifications (1.49 and 2.36 mm for the bionator and the RHS, respectively), with greater maxillary molar distalization in the RHS group. Overjet relationships also improved significantly compared with the controls (3.11 and 2.12 mm for the bionator and the RHS, respectively), due primarily to retroclination of the maxillary incisors (2.2 and 2.38 mm for the bionator and the RHS, respectively). The differences between overall corrections and dentoalveolar modifications for both molar and overjet relationships were explained by skeletal responses, with the bionator group showing significantly greater anterior mandibular displacement than the RHS group. Conclusions: The bionator and the RHS effectively corrected the molar relationships and overjets of Class II patients primarily by dentoalveolar changes. (Am J Orthod Dentofacial Orthop 2008; 134: 732-41)

Orthodontic movement after periodontal regeneration of class II furcation: a pilot study in dogs

Purpose: the effect of orthodontic movement on the periodontal tissues of maxillary second pre-molars, after regenerative treatment for class II furcations, was evaluated in four mongrel dogs.Material and Methods: Class II furcation lesions were created. After 75 days they were treated with bovine bone mineral matrix and guided tissue regeneration with absorbable membrane. After 2 months of daily plaque control, each of the dog's furcation pre-molars was randomly assigned to a test or control group. Orthodontic appliances were placed on both sides of the maxilla using third pre-molars and canines as anchorages. In the test group, bodily orthodontic movement of the second pre-molars was performed in the mesial direction for 3 months while control pre-molars remained unmoved. The dogs were sacrificed for histometric and histologic analyses.Results: There were no statistically significant differences between the two groups in total bone and biomaterial areas or linear extension of periodontal regeneration on the radicular surfaces. In the test group, however, there was a tendency to a greater quantity of bone and a lesser quantity of biomaterial.Conclusion: the orthodontic movement was not pre-judicial to the results obtained with the regenerative periodontal treatment.

Evaluation of marginal microleakage in Class II cavities: Effect of microhylolrid, flowable, and compactable resins

Objective: the goal of the present study was to evaluate the microleakage on the cementum/dentin and enamel surfaces in Class 11 restorations, using different kinds of resin composite (microhybrid, flowable, and compactable). Method and materials: Forty human caries-free molars were extracted and selected. Eighty Class 11 standardized cavities were made in the cervical wall at the cementoenamel junction (CEJ) and at the mesial and distal surfaces. The teeth were divided into four groups: G1 - adhesive system + microhybrid resin composite Z100; G2 - adhesive system + compactable resin composite Prodigy Condensable; G3 - adhesive system + flowable resin composite Revolution + Z1 00 resin composite; G4 - adhesive system + Revolution fluid resin + compactable resin composite Prodigy Condensable. The adhesive system used in this study was Scotchbond Multi-Purpose Plus. The specimens were thermocycled in baths of 5degreesC and 55degreesC for 1,000 cycles and immersed in 50% silver nitrate solution. The specimens then were sectioned and evaluated on degree of dye penetration. Results: the results were evaluated using the nonparametric Kruskall-Wallis test, which showed a statistically significant difference between groups G1 and G4, G2 and G4, and G3 and G4. Conclusions: None of the materials was able to eliminate the marginal microleakage at the cervical wall; the application of a low-viscosity resin composite combined with a compactable resin composite significantly decreased the microleakage.