Activation of Ca2+-independent membrane-damaging activity in Lys49-phospholipase A(2) promoted by amphiphilic molecules

Association of class-II phospholipase A(2) (PLA(2)) with aggregated phospholipid substrate results in elevated levels of the Ca2+-dependent hydrolytic activity. The Asp49 residue participates in coordination of the Ca2+ ion cofactor, however, in Lys49-PLA(2) homologues (Lys49-PLA(2)S), substitution of the Asp49 by Lys results in loss of Ca2+ binding and lack of detectable phospholipid hydrolysis. Nevertheless, Lys49-PLA2S cause Ca2+-independent damage of liposome membranes. Bothropstoxin-I is a homodimeric Lys49-PLA(2) from the venom of Bothrops jararacussu, and in fluorescent marker release and dynamic light scattering experiments with DPPC liposomes we demonstrate activation of the Ca2+-independent membrane damaging activity by similar to4 molecules of sodium dodecyl sulphate (SDS) per protein monomer. Activation is accomparlied by significant changes in the intrinsic tryptophan fluorescence emission (ITFE) and near UV circular dichroism (UVCD) spectra of the protein. Subsequent binding of 7-10 SDS molecules results in further alterations in the ITFE and far UVCD spectra. Reduction in the rate of N-bromosuccinimide modification of Trp77 at the dimer interface suggests that initial binding of SDS to this region accompanies the activation of the membrane damaging activity. 1-anilinonaphthalene-8-sulphonic acid binding studies indicate that subsequent SDS binding to the active site is concomitant with the second structural transition. These results provide insights in the structural basis of amphiphile/protein coupling in class-II PLA(2)s. (C) 2004 Published by Elsevier B.V.

Structural insights into the catalytic mechanism of sphingomyelinases D and evolutionary relationship to glycerophosphodiester phosphodiesterases

Spider venom sphingomyelinases D catalyze the hydrolysis of sphingomyelin via an Mg2+ ion-dependent acid-base catalytic mechanism which involves two histidines. In the crystal structure of the sulfate free enzyme determined at 1.85 angstrom resolution, the metal ion is tetrahedrally coordinated instead of the trigonal-bipyramidal coordination observed in the sulfate bound form. The observed hyperpolarized state of His47 requires a revision of the previously suggested catalytic mechanism. Molecular modeling indicates that the fundamental structural features important for catalysis are fully conserved in both classes of SMases D and that the Class II SMases D contain an additional intra-chain disulphide bridge (Cys53-Cys201). Structural analysis suggests that the highly homologous enzyme from Loxosceles bonetti is unable to hydrolyze sphingomyelin due to the 95G1y -> Asn and 134Pro -> Glu mutations that modify the local charge and hydrophobicity of the interfacial face. Structural and sequence comparisons confirm the evolutionary relationship between sphingomyelinases D and the glicerophosphodiester phosphoesterases which utilize a similar catalytic mechanism. (c) 2006 Elsevier B.V. All rights reserved.

Os efeitos dos aparelhos funcionais sobre a dimensão transversal da maxila e mandíbula

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Influence of the banded Herbst appliance on dental changes in mixed dentition

OBJECTIVE: This prospective clinical study was conducted with the purpose of evaluating the influence of the banded Herbst appliance on dental changes during the early treatment of Class II malocclusion. METHOD: The sample consisted of 15 prepubertal subjects (12 boys and 3 girls, initial age: 9 years and 6 months) who were treated with the Herbst appliance. Treatment effects were compared with those of a Class II Division 1 group of 15 subjects (8 boys and 7 girls, mean initial age 9 years and 1 month), not treated orthodontically. Statistical analysis was performed using Student t-test with 5% significance level. RESULTS: The results showed that treatment with the banded Herbst appliance in the mixed dentition stage tended to upright maxillary incisors (mean: 4.14°). The maxillary molars were distalized and intruded significantly (mean 2.65 mm and 1.24 mm, respectively), the lower incisors slightly protruded anteriorly (mean 1.64 mm) and the molars showed no significant changes in the horizontal and vertical directions. Furthermore, significant improvements were noted in overbite (1.26 mm), overjet (4.8 mm) and molar relationship (12.08 mm). CONCLUSIONS: Changes in the upper dental arch were found to be greater than changes in the lower arch. Furthermore, mandibular anchorage loss was reduced due to the anchorage system used in the study.

Cephalometric evaluation of skeletal alterations induced by Herbst appliance during mixed dentition

OBJECTIVE: The present prospective clinical study was designed in order to evaluate horizontal and vertical skeletal alterations induced by the use of Herbst appliance in individuals with Class II, division 1 malocclusion during mixed dentition stage. METHODS: The sampling consisted of 15 pre-pubertal individuals (12 boys and 3 girls; initial age 9 years and 6 months), who were treated with Herbst appliance for a period of 7 months. The effects of the treatment were compared to a group of 15 individuals with Class II, division 1 malocclusion (8 boys and 7 girls, initial age averaged 9 years and 1 month), orthodontically untreated, who were followed up for a period of 12 months. Statistical analysis was performed with Student's t-test with significance level at 5%. RESULTS: It was showed that the treatment with Herbst appliance in mixed dentition stage has restricted maxilla growth. Mandibular and palatal planes have not undergone significant alteration; however, anterior and posterior facial heights have increased significantly. Facial convexity and maxillomandibular relationship were altered positively. Mandible has positioned significantly forward and its effective length increased 2.5 times more than the increase observed in control group. CONCLUSION: It was possible to conclude that Herbst appliance was able to provide satisfactory results in individuals during mixed dentition stage.

Occurrence of skeletal malocclusions in Brazilian patients with dentofacial deformities

Este estudo avaliou a ocorrência de más oclusões esqueléticas apresentadas pelos pacientes do Centro de Pesquisa e Tratamento das Deformidades Bucofaciais (CEDEFACE), na cidade de Araraquara, SP, Brasil. Foram avaliados prontuários de 381 pacientes com deformidades dentoesqueléticas, que fizeram tratamento combinado ortodôntico-cirúrgico no período entre 2000 e 2006. Após a seleção da amostra (método de conveniência), baseado nos dados da documentação pré e pós-cirúrgica, o número de pacientes foi reduzido para 171. Para classificação do levantamento, considerou-se a discrepância ântero-posterior (Classe I, II e III), raça, idade, gênero, ausência ou presença de assimetria, excesso vertical maxilar e biprotrusão maxilar, além de determinar em qual base óssea o procedimento cirúrgico foi realizado. As documentações dos pacientes foram analisadas por um examinador previamente calibrado pelo processo de repetição até que o método fosse considerado adequado (correlação intraclasse >0,94). A idade média dos pacientes foi de 23,59 anos (DP 6,93), a maioria do gênero feminino (102 pacientes) e leucoderma (160 pacientes). A má oclusão mais prevalente foi a Classe III (81 pacientes). A assimetria, o excesso maxilar vertical e biprotrusão maxilar estavam presentes em 54, 33, e 7 pacientes, respectivamente. Na maioria dos casos, as cirurgias para correção de deformidades dentoesqueléticas foram combinadas, envolvendo os dois maxilares. Com base nos resultados, conclui-se que a Classe III foi a deformidade esquelética mais prevalente e a Classe I a menos prevalente. em geral, a prevalência de deformidades esqueléticas foi maior entre as mulheres e a maioria dos pacientes apresentou uma combinação de problemas maxilares e mandibulares, o que interfere diretamente na decisão sobre o plano de tratamento mais adequado. Houve uma maior incidência de assimetria na Classe III esquelética; o excesso vertical ocorreu de forma semelhante na Classe II e III e a biprotrusão teve baixa incidência entre as más oclusões avaliadas.

Efeitos dentoesqueléticos produzidos pelo aparelho de Herbst na dentadura mista

OBJETIVO: o objetivo desta pesquisa clínica prospectiva foi avaliar as alterações cefalométricas dentárias e esqueléticas produzidas pelo aparelho de Herbst em jovens com má oclusão de Classe II, 1ª divisão durante a dentadura mista. METODOLOGIA: trinta jovens (15 do gênero masculino e 15 do feminino) com idade média inicial de 9 anos e 10 meses foram tratados com o aparelho de Herbst por um período de 12 meses. Para a comparação dos grupos utilizou-se uma amostra controle de 30 jovens (15 do gênero masculino e 15 do feminino) Classe II, 1ª divisão, com idade média inicial de 9 anos e 8 meses, que foram mantidos sem tratamento durante 12 meses. Para cada jovem foram utilizadas duas telerradiografias em norma lateral, obtidas ao início e no final do período de acompanhamento. Utilizou-se um método convencional de avaliação cefalométrica e o método proposto por Pancherz. RESULTADOS E CONCLUSÕES: os resultados deste estudo demonstraram que os efeitos do aparelho de Herbst produzidos na dentadura mista foram primariamente de natureza dentoalveolar. Os incisivos inferiores foram inclinados para vestibular e os superiores foram retruídos; também houve uma extrusão significante dos molares inferiores, enquanto os superiores sofreram restrição de desenvolvimento no sentido vertical. Não houve diferença significante de restrição do crescimento anterior da maxila entre os dois grupos. No sentido vertical da face, a altura facial ântero-inferior se comportou de forma similar, não demonstrando alteração significante entre os grupos. O tratamento com o aparelho de Herbst produziu um aumento modesto, porém, significante no comprimento da mandíbula comparado ao grupo controle. Este aumento, entretanto, foi de menor magnitude que aquele observado em pacientes adolescentes utilizando o mesmo protocolo de tratamento. A correção do overjet (Herbst) ocorreu devido a 22% de alterações esqueléticas e 78% de alterações dentárias. A correção da relação molar ocorreu devido a 27% de alterações esqueléticas e 73% de alterações dentárias.

Antitumor effects in vitro and in vivo and mechanisms of protection against melanoma B16F10-Nex2 cells by fastuosain, a cysteine proteinase from Bromelia fastuosa

In the present work, the antitumor effect of fastuosain, a cysteine proteinase from Bromelia fastuosa, was investigated. In the intravenous model of lung colonization in C57Bl/6 mice, fastuosain and bromelain injected intraperitoneally were protective, and very few nodules of B16F10-Nex2 melanoma cells were detected. Tumor cells treated with fastuosain showed reduced expression of CD44 and decreased invasion through Matrigel, lost their cytoplasmic extensions and substrate adherence, and became round and detached, forming strongly bound cell clusters in suspension. Peritoneal cells recruited and activated by fastuosain treatment ( mainly monocytic cells and lymphocytes) migrated to the lung, where pulmonary melanoma metastases grew. Adoptive transference of peritoneal cells recruited by fastuosain had no protective effect against lung metastases in recipient mice. Treatment of green fluorescent protein - chimeric animals with fastuosain did not change the number of cells that migrated to the lung, compared to PBS-injected control mice, but the number of positive major histocompatibility complex class II cells increased with fastuosain treatment. Murine antibodies against fastuosain, bromelain, and cathepsins B and L cross-reacted in ELISA and recognized surface and cytoplasmic components expressed on B16F10-Nex2 cells. Anti-fastuosain antibodies were cytotoxic/lytic to B16F10-Nex2 cells. Antitumor effects of fastuosain involve mainly the direct effect of the enzyme and elicitation of protective antibodies.

Transverse skeletal base adaptations with Bionator therapy: A pilot implant study

The purpose of this randomized, controlled trial was to evaluate transverse skeletal base adaptations to Bionator therapy. The sample included 25 patients (15 male, 10 female) aged 6.9 to 11.2 years with Class II Division 1 malocclusion. The patients were randomly allocated to either a control (n = 11) or treatment (n = 14) group and followed longitudinally for approximately 12 months. Treatment consisted of a Bionator only, constructed to remain approximately 2 mm from the buccal dentition. Transverse maxillary and mandibular changes were evaluated cephalometrically according to 4 bilateral maxillary and 2 bilateral mandibular implants. Untreated Class II controls exhibited significant increases between posterior maxillary implants but no significant changes between the anterior maxillary or mandibular implants. There were no significant width differences between the control and treated groups before treatment. Posterior maxillary implant widths increased significantly (P < .05) in both groups, but the treated group showed significantly greater width increases than the control group. The treated group also showed greater increases between mandibular implants, but the differences were not statistically significant. These results suggest that transverse skeletal base adaptations occur as a result of Bionator therapy.

Analysis of the inflammatory reaction induced by the catfish (Cathorops spixii) venoms

Cathorops spixii is one of the most abundant venomous fish of the southeastern coast of the State of São Paulo, and consequently causes a great part of the accidents seen there. The accidents affect mainly fishermen, swimmers and tourists and are characterized by punctiform or wide wounds, erythema, edema, pain, sudoresis, indisposition, fever, nausea, vomiting and secondary infection. The objective of this work was to characterize the inflammatory response induced in mice by both venoms (mucus and sting) of the catfish C spixii. Our results demonstrated that both venoms induced a great number of rolling and adherent leukocytes in the post-capillary venules of cremaster muscle of mice, and an increase in the vascular permeability in peritoneal cavity. Mucus induced the recruitment of neutrophils immediately after injection followed later by macrophage infiltration. In contrast, the cellular infiltration elicited by sting venom was rapidly resolved. The peritonitis reaction provoked by venoms was characterized by cytokine (IL-6), chemokines (MCP-1 and KC) or lipid mediator (LTB4) production in the peritoneal cavity. The macrophages from 7-day mucus venom-induced exudates upon in vitro mucus venom stimulation, expressed CD1 Ic x MHC class II and release bioactive IL-12p70. on the other hand, sting venom-elicited peritoneal macrophages lost the ability to differentiate into dendritic cells, following re-stimulation in vitro with sting venom, they do not express CD11c, nor do they exhibit sufficient levels of MHC class II. In conclusion, both types of venoms (mucus or sting) promote inflammatory reaction with different profiles, and the inflammatory reaction induced by the first was characterized by antigen persistence in peritoneal cavity that allowed the activation of phagocytic cells with capacity of antigenic presentation. (C) 2007 Elsevier Ltd. All rights reserved.

Long-term effectiveness and treatment timing for bionator therapy

The aim of the present investigation was to provide information about the long-term effects and optimal timing for class-II treatment with the Bionator appliance. Lateral cephalograms of 23 class-II patients treated with the Bionator were analyzed at three time periods: T1, start of treatment; T2, end of Bionator therapy; and T3, long-term observation (after completion of growth). T3 includes a phase with fixed appliances. The treated sample was divided into two groups according to their skeletal maturity as evaluated by the cervical vertebral maturation (CVM) method. The early-treated group (13 subjects) initiated treatment before the peak in mandibular growth, which occurred after completion of Bionator therapy. The late-treated group (10 subjects) received Bionator treatment during the peak. The T1-T2, T2-T3, and T1-T3 changes in the treated groups were compared with changes in control groups of untreated class-II subjects by nonparametric statistics (P < .05). The findings of the present study on Bionator therapy followed by fixed appliances indicate that this treatment protocol is more effective and stable when it is performed during the pubertal growth spurt. Optimal timing to start treatment with the Bionator is when a concavity appears at the lower borders of the second and the third cervical vertebrae (CVMS 11). In the long term, the amount of significant supplementary elongation of the mandible in subjects treated during the pubertal peak is 5.1 mm more than in the controls, and it is associated with a backward direction of condylar growth. Significant increments in mandibular ramus height also were recorded.

Protective Effect of HLA-DRB1*11 and Predisposition of HLA-C*04 in the Development of Severe Liver Damage in Brazilian Patients with Chronic Hepatitis C Virus Infection

The objective of this study was to investigate human leucocyte antigen (HLA) genes in patients chronically infected with hepatitis C virus (HCV) and to analyse the possible role of these genes in the progression of chronic hepatitis C. One hundred and forty-five (145) Brazilian patients infected only with HCV genotype 1 were evaluated. HLA class I (A*, B*, C*) and class II (DRB1*, DQA1*, DQB1*) typing were carried out by PCR-SSO, through Luminex technology. Associations were found with protection against development of liver damage by both DRB1*11 (5.0% versus 18.2%, P = 0.0016, OR = 0.23, CI 95% = 0.090.58; Pc=0.0208) and DRB1*11-DQA1*05-DQB1*03 haplotype (4.2% versus 15.3%, P = 0.0032; OR = 0.24, CI 95% = 0.08-0.64). Liver damage was associated with HLA-C*04 in patients with <20 years of infection (38.4% versus 9.1%, P = 0.002, OR = 6.25, CI 95% = 1.9719.7; Pc=0.0238). It is concluded that HLA alleles can influence the development of liver damage in HCV type-1 chronically infected Brazilian patients.

Influence of HLA alleles in response to treatment with pegylated interferon-alpha and ribavirin in patients with chronic hepatitis C

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Estudo da infiltração marginal em restaurações de resinas compostas para dentes posteriores. Efeito do material, preparo cavitário e condicionamento do esmalte a nível cervical.

The purpose of this in vitro study was to investigate the cervical marginal leakage in class II restorations with chemically cured resin (P10) and light-cured resin (P30) in two types of cavities: conventional and adhesive. The effect of acid-etching in this area was also observed. Dentine adhesive Scotchbond was used in all experimental groups. Leakage was evidenced by Rodamina B dye penetration after thermocycling procedure between 10 degrees C and 50 degrees C temperature and analysed by using Zeiss Stereoscopic Magnifying Glass (10 X). According to the results obtained marginal leakage occurred in all experimental groups, with lower percentage for adhesives cavities when enamel acid-etching and light-cured resin P30 was used.

Effects of cervical headgear and edgewise appliances on growing patients

Maxillary basal bone, dentoalveolar, and dental changes in Class II Division 1 patients treated to normal occlusion by using cervical headgear and edgewise appliances were retrospectively evaluated. A sample of 45 treated patients was compared with a group of 30 untreated patients. Subjects were drawn from the Department of Orthodontics, Araraquara School of Dentistry, Brazil, and ranged in age from 7.5 to 13.5 years. The groups were matched based on age, gender, and malocclusion. Roughly 87% of the treated group had a mesocephalic or brachicephalic pattern, and 13% had a dolicocephalic pattern. Cervical headgear was used until a Class I dental relationship was achieved. Our results demonstrated that the malocclusions were probably corrected by maintaining the maxillary first molars in position during maxillary growth. Maxillary basal bone changes (excluding dentoalveolar changes) did not differ significantly between the treated and the untreated groups. Molar extrusion after the use of cervical headgear was not supported by our data, and this must be considered in the treatment plan of patients who present similar facial types. (Am J Orthod Dentofacial Orthop 2001;119:531-9).