AVALIAÇÃO CEFALOMÉTRICA DA ESTABILIDADE PÓS-EXPANSÃO RÁPIDA DA MAXILA ASSISTIDA CIRURGICAMENTE

Este estudo avaliou a estabilidade das alterações dentárias e esqueléticas produzidas pela Expansão Rápida da Maxila Assistida Cirurgicamente (ERMAC), no sentido transversal e vertical. A amostra selecionada para este estudo retrospectivo foi composta de 60 telerradiografias em norma frontal, de 15 pacientes, sendo 6 do sexo masculino e 9 do sexo feminino, com média de idade de 23 anos e 3 meses. Utilizou-se o disjuntor tipo Hyrax e o procedimento cirúrgico foi caracterizado pela osteotomia sagital mediana da maxila e não abordagem da sutura pterigopalatina. O início da ativação ocorreu no terceiro dia pós-operatório, sendo que, os limites para a expansão foram determinados por critérios eminentemente clínicos. Todos os pacientes foram radiografados nas fases pré-expansão (T1), pós-expansão imediata (T2), 3 meses pós- expansão (com o próprio disjuntor como contenção) (T3) e 6 meses pós-expansão (com a placa de acrílico removível como contenção) (T4). Medidas lineares foram obtidas a partir dos traçados cefalométricos gerados por um programa computadorizado (Radiocef Studio 2) e analisadas estatisticamente pelo teste de variância (ANOVA) e Tukey ao nível de 5% de significância. Concluiu-se que a ERMAC produziu um aumento estatisticamente significante, da cavidade nasal, largura maxilar, distância intermolares superiores, de T1 para T2, e que se mantiveram em T3 e T4. A largura facial e as distâncias intermolares inferiores não apresentaram alterações após a ERMAC. Avaliando o comportamento vertical da face, notou-se um aumento da AFAI nos tempos T1 para T2 que, diminuiu após a contenção de 3 meses (T3) e permaneceu estável em T4, embora aumentada se comparada com T1.(AU)

Avaliação da influência dos formatos DICOM e JPEG na reprodutibilidade de pontos cefalométricos em Telerradiografia digital em Norma Frontal

A imagem digital no formato DICOM requer grande espaço para armazenamento, dificultando o arquivamento e transmissão da imagem via internet, sendo necessária, muitas vezes, a compressão das imagens por meio de formatos de arquivo como o JPEG. O objetivo neste estudo foi avaliar a influência dos formatos DICOM e JPEG, nos Fatores de Qualidade 100, 80 e 60, na reprodutibilidade intra e interexaminador na marcação de pontos cefalométricos em Telerradiografias digitais em Norma Frontal. A amostra consistiu de 120 imagens digitais de Telerradiografias em Norma Frontal, obtidas de 30 indivíduos. As 30 imagens originais, em formato DICOM, posteriormente, foram convertidas para o formato JPEG, nos Fatores de Qualidade 100, 80 e 60. Após cegar e randomizar a amostra, três ortodontistas calibrados marcaram os 18 pontos cefalométricos em cada imagem utilizando um programa de cefalometria computadorizada, que registra as medidas dos pontos cefalométricos em um sistema de coordenadas cartesianas X e Y. Nos resultados, os testes estatísticos de correlações intraclasses e análise de variância (ANOVA) apresentaram concordância de reprodutibilidade dos pontos cefalométricos em Telerradiografias digitais em Norma Frontal, tanto intra como interexaminador, com exceção dos pontos ZL, ZR, AZ, JR, NC, CN na coordenada Y e A6 na coordenada X, independentemente dos formatos de arquivo. Em conclusão, os formatos de arquivo DICOM e JPEG, nos Fatores de Qualidade 100, 80 e 60, não afetaram a reprodutibilidade intra e interexaminador na marcação dos pontos cefalométricos.(AU)

Avaliação da influência dos formatos DICOM e JPEG na reprodutibilidade de pontos cefalométricos em Telerradiografia digital em Norma Frontal

A imagem digital no formato DICOM requer grande espaço para armazenamento, dificultando o arquivamento e transmissão da imagem via internet, sendo necessária, muitas vezes, a compressão das imagens por meio de formatos de arquivo como o JPEG. O objetivo neste estudo foi avaliar a influência dos formatos DICOM e JPEG, nos Fatores de Qualidade 100, 80 e 60, na reprodutibilidade intra e interexaminador na marcação de pontos cefalométricos em Telerradiografias digitais em Norma Frontal. A amostra consistiu de 120 imagens digitais de Telerradiografias em Norma Frontal, obtidas de 30 indivíduos. As 30 imagens originais, em formato DICOM, posteriormente, foram convertidas para o formato JPEG, nos Fatores de Qualidade 100, 80 e 60. Após cegar e randomizar a amostra, três ortodontistas calibrados marcaram os 18 pontos cefalométricos em cada imagem utilizando um programa de cefalometria computadorizada, que registra as medidas dos pontos cefalométricos em um sistema de coordenadas cartesianas X e Y. Nos resultados, os testes estatísticos de correlações intraclasses e análise de variância (ANOVA) apresentaram concordância de reprodutibilidade dos pontos cefalométricos em Telerradiografias digitais em Norma Frontal, tanto intra como interexaminador, com exceção dos pontos ZL, ZR, AZ, JR, NC, CN na coordenada Y e A6 na coordenada X, independentemente dos formatos de arquivo. Em conclusão, os formatos de arquivo DICOM e JPEG, nos Fatores de Qualidade 100, 80 e 60, não afetaram a reprodutibilidade intra e interexaminador na marcação dos pontos cefalométricos.(AU)

Comunicação social com software livre

A tecnologização que a sociedade experimenta nas últimas décadas trouxe a profusão de máquinas informatizadas e seus sistemas de operação. Neste período a indústria desenvolveu sofisticados e caros softwares aplicativos proprietários para o pleno uso destas máquinas, o que colocou boa parte do mercado social nas mãos de poucas empresas multinacionais, entre elas, a Microsoft, e outras. Mas, o espírito libertário de membros das comunidades científicas e hackers promoveu o desenvolvimento do software livre e aberto, que pode ser usado como bem social mais amplo e, principalmente, evoluir no melhor do espírito colaborativo. O presente trabalho estuda os dois modelos de produção de software, os compara visando tornar evidentes as qualidades de cada um, seus custos, rendimentos e possibilidades de adoção. Projeta a possibilidade de que as habilitações da área da comunicação possam migrar para o modelo de software livre, dadas as plenas qualidades deste sistema, a radical redução de custos e as constatações que amplos segmentos da produção audiovisual os está adotando. Para tanto, compara as experiências aplicadas com ambos os sistemas em dois cursos de comunicação, em sua habilitação de Rádio e Televisão.(AU)

Attenuation of virulence in Mycobacterium tuberculosis expressing a constitutively active iron repressor

Iron is an essential nutrient for the survival of most organisms and has played a central role in the virulence of many infectious disease pathogens. Mycobacterial IdeR is an iron-dependent repressor that shows 80% identity in the functional domains with its corynebacterial homologue, DtxR (diphtheria toxin repressor). We have transformed Mycobacterium tuberculosis with a vector expressing an iron-independent, positive dominant, corynebacterial dtxR hyperrepressor, DtxR(E175K). Western blots of whole-cell lysates of M. tuberculosis expressing the dtxR(E175K) gene revealed the stable expression of the mutant protein in mycobacteria. BALB/c mice were infected by tail vein injection with 2 × 105 organisms of wild type or M. tuberculosis transformed with the dtxR mutant. At 16 weeks, there was a 1.2 log reduction in bacterial survivors in both spleen (P = 0.0002) and lungs (P = 0.006) with M. tuberculosis DtxR(E175K). A phenotypic difference in colonial morphology between the two strains also was noted. A computerized search of the M. tuberculosis genome for the palindromic consensus sequence to which DtxR and IdeR bind revealed six putative “iron boxes” within 200 bp of an ORF. Using a gel-shift assay we showed that purified DtxR binds to the operator region of five of these boxes. Attenuation of M. tuberculosis can be achieved by the insertion of a plasmid containing a constitutively active, iron-insensitive repressor, DtxR(E175K), which is a homologue of IdeR. Our results strongly suggest that IdeR controls genes essential for virulence in M. tuberculosis.

Adeno-associated viral-mediated catalase expression suppresses optic neuritis in experimental allergic encephalomyelitis

Suppression of oxidative injury by viral-mediated transfer of the human catalase gene was tested in the optic nerves of animals with experimental allergic encephalomyelitis (EAE). EAE is an inflammatory autoimmune disorder of primary central nervous system demyelination that has been frequently used as an animal model for the human disease multiple sclerosis (MS). The optic nerve is a frequent site of involvement common to both EAE and MS. Recombinant adeno-associated virus containing the human gene for catalase was injected over the right optic nerve heads of SJL/J mice that were simultaneously sensitized for EAE. After 1 month, cell-specific catalase activity, evaluated by quantitation of catalase immunogold, was increased approximately 2-fold each in endothelia, oligodendroglia, astrocytes, and axons of the optic nerve. Effects of catalase on the histologic lesions of EAE were measured by computerized analysis of the myelin sheath area (for demyelination), optic disc area (for optic nerve head swelling), extent of the cellular infiltrate, extravasated serum albumin labeled by immunogold (for blood–brain barrier disruption), and in vivo H2O2 reaction product. Relative to control, contralateral optic nerves injected with the recombinant virus without a therapeutic gene, catalase gene inoculation reduced demyelination by 38%, optic nerve head swelling by 29%, cellular infiltration by 34%, disruption of the blood–brain barrier by 64%, and in vivo levels of H2O2 by 61%. Because the efficacy of potential treatments for MS are usually initially tested in the EAE animal model, this study suggests that catalase gene delivery by using viral vectors may be a therapeutic strategy for suppression of MS.

MEKK1/JNK signaling stabilizes and activates p53

Activation of the tumor suppressor p53 by stress and damage stimuli often correlates with induction of stress kinases, Jun-NH2 kinase (JNK). As JNK association with p53 plays an important role in p53 stability, in the present study we have elucidated the relationship between the JNK-signaling pathway and p53 stability and activity. Expression of a constitutively active form of JNKK upstream kinase, mitogen-activated protein kinase kinase kinase (ΔMEKK1), increased the level of the exogenously transfected form of p53 in p53 null (10.1) cells as well as of endogenous p53 in MCF7 breast cancer cells. Increased p53 level by forced expression of ΔMEKK1 coincided with a decrease in p53 ubiquitination in vivo and with prolonged p53 half-life. Computerized modeling of the JNK-binding site (amino acids 97–116; p7 region) enabled us to design mutations of exposed residues within this region. Respective mutations (p53101-5-8) and deletion (p53Δp7) forms of p53 did not exhibit the same increase in p53 levels upon ΔMEKK1 expression. In vitro phosphorylation of p53 by JNK abolished Mdm2 binding and targeting of p53 ubiquitination. Similarly, ΔMEKK1 expression increased p53 phosphorylation by immunopurified JNK and dissociated p53–Mdm2 complexes. Transcriptional activity of p53, as measured via mdm2 promoter-driven luciferase, exhibited a substantial increase in ΔMEKK1-expressing cells. Cotransfection of p53 and ΔMEKK1 into p53 null cells potentiated p53-dependent apoptosis, suggesting that MEKK1 effectors contribute to the ability of p53 to mediate programmed cell death. Our results point to the role of MEKK1-JNK signaling in p53 stability, transcriptional activities, and apoptotic capacity as part of the cellular response to stress.

PAGE-1, an X chromosome-linked GAGE-like gene that is expressed in normal and neoplastic prostate, testis, and uterus

We have used a combination of computerized database mining and experimental expression analyses to identify a gene that is preferentially expressed in normal male and female reproductive tissues, prostate, testis, fallopian tube, uterus, and placenta, as well as in prostate cancer, testicular cancer, and uterine cancer. This gene is located on the human X chromosome, and it is homologous to a family of genes encoding GAGE-like proteins. GAGE proteins are expressed in a variety of tumors and in testis. We designate the novel gene PAGE-1 because the expression pattern in the Cancer Genome Anatomy Project libraries indicates that it is predominantly expressed in normal and neoplastic prostate. Further database analysis indicates the presence of other genes with high homology to PAGE-1, which were found in cDNA libraries derived from testis, pooled libraries (with testis), and in a germ cell tumor library. The expression of PAGE-1 in normal and malignant prostate, testicular, and uterine tissues makes it a possible target for the diagnosis and possibly for the vaccine-based therapy of neoplasms of prostate, testis, and uterus.

Distinct gene-specific mechanisms of arrhythmia revealed by cardiac gene transfer of two long QT disease genes, HERG and KCNE1

The long QT syndrome (LQTS) is a heritable disorder that predisposes to sudden cardiac death. LQTS is caused by mutations in ion channel genes including HERG and KCNE1, but the precise mechanisms remain unclear. To clarify this situation we injected adenoviral vectors expressing wild-type or LQT mutants of HERG and KCNE1 into guinea pig myocardium. End points at 48–72 h included electrophysiology in isolated myocytes and electrocardiography in vivo. HERG increased the rapid component, IKr, of the delayed rectifier current, thereby accelerating repolarization, increasing refractoriness, and diminishing beat-to-beat action potential variability. Conversely, HERG-G628S suppressed IKr without significantly delaying repolarization. Nevertheless, HERG-G628S abbreviated refractoriness and increased beat-to-beat variability, leading to early afterdepolarizations (EADs). KCNE1 increased the slow component of the delayed rectifier, IKs, without clear phenotypic sequelae. In contrast, KCNE1-D76N suppressed IKs and markedly slowed repolarization, leading to frequent EADs and electrocardiographic QT prolongation. Thus, the two genes predispose to sudden death by distinct mechanisms: the KCNE1 mutant flagrantly undermines cardiac repolarization, and HERG-G628S subtly facilitates the genesis and propagation of premature beats. Our ability to produce electrocardiographic long QT in vivo with a clinical KCNE1 mutation demonstrates the utility of somatic gene transfer in creating genotype-specific disease models.

A methylated human 9-kb repetitive sequence on acrocentric chromosomes is homologous to a subtelomeric repeat in chimpanzees.

We have implemented an approach for the detection of DNA alterations in cancer by means of computerized analysis of end-labeled genomic fragments, separated in two dimensions. Analysis of two-dimensional patterns of neuroblastoma tumors, prepared by first digesting DNA with the methylation-sensitive restriction enzyme Not I, yielded a multicopy fragment which was detected in some tumor patterns but not in normal controls. Cloning and sequencing of the fragment, isolated from two-dimensional gels, yielded a sequence with a strong homology to a subtelomeric sequence in chimpanzees and which was previously reported to be undetectable in humans. Fluorescence in situ hybridization indicated the occurrence of this sequence in normal tissue, for the most part in the satellite regions of acrocentric chromosomes. A product containing this sequence was obtained by telomere-anchored PCR using as a primer an oligonucleotide sequence from the cloned fragment. Our data suggest demethylation of cytosines at the cloned Not I site and in neighboring DNA in some tumors, compared with normal tissue, and suggest a greater similarity between human and chimpanzee subtelomeric sequences than was previously reported.

Vestibular navigation directed by the slope of terrain.

Slope of terrain is an important orienting gradient affecting the goal-directed locomotion of animals. Its significance was assessed in experiment 1 by training rats to find in darkness a feeder on the top of a low cone (80-cm base, 0- to 4-cm high). A computerized infrared tracking system monitoring the rat's position in darkness showed that the path length on the cone surface was inversely proportional to cone height. A device allowing continuous generation of slope-guided locomotion was used in experiment 2. This device consists of a 1-m arena, the floor of which can be supported at a point corresponding to the position of one of three equidistant feeders located 17 cm from its center. The arena is inclined by the locomotion of the rat to a plane passing through the elevated (2- or 4-cm) feeder, the rat's center of gravity, and a point at the edge of the arena resting on the floor. The multitude of such planes generated by the rat's locomotion forms the surface of a virtual cone, the top of which is formed by the feeder. Additional path (difference between distance traveled and shortest distance of the animal from the goal at the onset of inclination) is inversely related to the incline of the arena and is a sensitive measure of performance in this type of vestibular navigation.

Peptides from conserved regions of paramyxovirus fusion (F) proteins are potent inhibitors of viral fusion.

The synthetic peptides DP-107 and DP-178 (T-20), derived from separate domains within the human immunodeficiency virus type 1 (HIV-1) transmembrane (TM) protein, gp4l, are stable and potent inhibitors of HIV-1 infection and fusion. Using a computer searching strategy (computerized antiviral searching technology, C.A.S.T.) based on the predicted secondary structure of DP-107 and DP-178 (T-20), we have identified conserved heptad repeat domains analogous to the DP-107 and DP-178 regions of HIV-1 gp41 within the glycoproteins of other fusogenic viruses. Here we report on antiviral peptides derived from three representative paramyxoviruses, respiratory syncytial virus (RSV), human parainfluenza virus type 3 (HPIV-3), and measles virus (MV). We screened crude preparations of synthetic 35-residue peptides, scanning the DP-178-like domains, in antiviral assays. Peptide preparations demonstrating antiviral activity were purified and tested for their ability to block syncytium formation. Representative DP-178-like peptides from each paramyxovirus blocked homologous virus-mediated syncytium formation and exhibited EC50 values in the range 0.015-0.250 microM. Moreover, these peptides were highly selective for the virus of origin. Identification of biologically active peptides derived from domains within paramyxovirus F1 proteins analogous to the DP-178 domain of HIV-1 gp4l is compelling evidence for equivalent structural and functional features between retroviral and paramyxoviral fusion proteins. These antiviral peptides provide a novel approach to the development of targeted therapies for paramyxovirus infections.

Rapid mass spectrometric peptide sequencing and mass matching for characterization of human melanoma proteins isolated by two-dimensional PAGE.

We report a general mass spectrometric approach for the rapid identification and characterization of proteins isolated by preparative two-dimensional polyacrylamide gel electrophoresis. This method possesses the inherent power to detect and structurally characterize covalent modifications. Absolute sensitivities of matrix-assisted laser desorption ionization and high-energy collision-induced dissociation tandem mass spectrometry are exploited to determine the mass and sequence of subpicomole sample quantities of tryptic peptides. These data permit mass matching and sequence homology searching of computerized peptide mass and protein sequence data bases for known proteins and design of oligonucleotide probes for cloning unknown proteins. We have identified 11 proteins in lysates of human A375 melanoma cells, including: alpha-enolase, cytokeratin, stathmin, protein disulfide isomerase, tropomyosin, Cu/Zn superoxide dismutase, nucleoside diphosphate kinase A, galaptin, and triosephosphate isomerase. We have characterized several posttranslational modifications and chemical modifications that may result from electrophoresis or subsequent sample processing steps. Detection of comigrating and covalently modified proteins illustrates the necessity of peptide sequencing and the advantages of tandem mass spectrometry to reliably and unambiguously establish the identity of each protein. This technology paves the way for studies of cell-type dependent gene expression and studies of large suites of cellular proteins with unprecedented speed and rigor to provide information complementary to the ongoing Human Genome Project.

Quantitative analysis of senile plaques in Alzheimer disease: observation of log-normal size distribution and molecular epidemiology of differences associated with apolipoprotein E genotype and trisomy 21 (Down syndrome).

The discovery that the epsilon 4 allele of the apolipoprotein E (apoE) gene is a putative risk factor for Alzheimer disease (AD) in the general population has highlighted the role of genetic influences in this extremely common and disabling illness. It has long been recognized that another genetic abnormality, trisomy 21 (Down syndrome), is associated with early and severe development of AD neuropathological lesions. It remains a challenge, however, to understand how these facts relate to the pathological changes in the brains of AD patients. We used computerized image analysis to examine the size distribution of one of the characteristic neuropathological lesions in AD, deposits of A beta peptide in senile plaques (SPs). Surprisingly, we find that a log-normal distribution fits the SP size distribution quite well, motivating a porous model of SP morphogenesis. We then analyzed SP size distribution curves in genotypically defined subgroups of AD patients. The data demonstrate that both apoE epsilon 4/AD and trisomy 21/AD lead to increased amyloid deposition, but by apparently different mechanisms. The size distribution curve is shifted toward larger plaques in trisomy 21/AD, probably reflecting increased A beta production. In apoE epsilon 4/AD, the size distribution is unchanged but the number of SP is increased compared to apoE epsilon 3, suggesting increased probability of SP initiation. These results demonstrate that subgroups of AD patients defined on the basis of molecular characteristics have quantitatively different neuropathological phenotypes.

Gerenciamento integrado de resíduos sólidos: estudo de caso no Instituto Butantan

A relevância do gerenciamento de resíduos sólidos justifica-se pelos impactos à saúde e ao meio ambiente. Resíduos gerados em serviços de saúde podem apresentar periculosidade por suas características físico-químicas e biológicas. No Brasil, todo grande gerador é obrigado a elaborar o Plano de Gerenciamento de Resíduos Sólidos, e todo estabelecimento de saúde deve criar o Plano de Gerenciamento de Resíduos de Serviços de Saúde. Objetivo: Desenvolver modelo de gestão de resíduos sólidos para apoiar a elaboração e implantação desses planos em instituições públicas de pesquisa, desenvolvimento e fabricação de produtos de saúde. Métodos: Estudo de caso conduzido no Instituto Butantan, localizado no município de São Paulo/SP. Foi realizado considerando as seguintes etapas: diagnóstico do gerenciamento dos resíduos e elaboração, implantação e avaliação de Plano Integrado de Gerenciamento de Resíduos Sólidos. Todo o processo teve como abordagem a gestão participativa, compartilhada e integrada, envolvendo todos os atores da instituição. Na avaliação foram considerados o atendimento legal quanto à gestão e gerenciamento, às práticas e procedimentos implantados e à atuação dos envolvidos. Resultados: Destacam-se a caracterização e quantificação para cada fluxo de resíduos, a elaboração do Guia Prático de Descarte de Resíduos, o Modelo de Gestão para instituições de pesquisa, desenvolvimento e fabricação de produtos de uso em saúde humana baseado nas premissas do ciclo PDCA e o sistema informatizado de gerenciamento para estruturação e controle da gestão dos resíduos. Conclusões: Verifica-se que o processo desenvolvido ao longo do estudo propiciou mudança de cultura, envolvimento dos funcionários por meio da capacitação contínua para atuação e segurança do trabalhador e melhoria no gerenciamento dos resíduos, promovendo a redução de custos de destinação e a valorização dos resíduos.