Efeito do laser de baixa intensidade no procedimento de expansão rápida da maxila

A proposta do presente estudo foi avaliar os efeitos do laser de baixa intensidade na regeneração óssea no procedimento de expansão rápida da maxila. Utilizou-se 27 indivíduos com média de idade de 10,2 anos, divididos em dois grupos: grupo laser (n=14), no qual se realizou a expansão rápida da maxila, associada ao laser e grupo sem laser (n=13), que realizou somente a expansão rápida da maxila. O protocolo de ativação do parafuso expansor foi de 1 volta completa no primeiro dia e ½ volta diária até a sobrecorreção. O laser utilizado foi o de diodo (TWIN Laser MMOptics®, São Carlos), seguindo o protocolo de aplicação: comprimento de onda de 780nm, potência de 40mW, densidade de 10J/cm2, em 10 pontos localizados ao redor da sutura palatina mediana. Os estágios de aplicação foram: L1 (do primeiro ao quinto dia de aplicação), L2 (travamento do parafuso e 3 dias seguidos), L3, L4 e L5 (após 7, 14 e 21 dias do L2, respectivamente). Radiografias oclusais da maxila foram realizadas com auxílio de uma escala de alumínio, para referencial densitométrico, em diferentes tempos: T1 (inicial), T2 (dia de travamento do parafuso), T3 (3 a 5 dias do T2), T4 (30 dias do T3), T5 (60 dias do T4). As radiografias foram digitalizadas e submetidas a um programa de imagem (Image Tool - UTHSCSA, Texas, USA), para mensuração da densidade óptica das áreas previamente selecionadas. Para realização do teste estatístico, utilizou-se a Análise de Covariância usando como covariável o tempo para a fase avaliada. Em todos os testes foi adotado nível de significância de 5% (p<0,05).Para o Grupo Laser, os dados mostram que houve uma queda significante de densidade durante a abertura do parafuso (T2-T1), um aumento significante da mesma no período final de avaliação (T5-T4), e um aumento também da densidade no período de regeneração propriamente dito (T5-T2), ou seja, a partir do momento em que finalizou a fase de abertura do parafuso expansor. Enquanto que no Grupo Sem Laser, a densidade não mostrou diferença estatisticamente significantemente em nenhum período analisado. Os resultados mostraram que o laser propiciou consideravelmente uma melhor abertura da sutura palatina mediana, além de influenciar no processo de regeneração óssea da sutura, acelerando seus processos de reparo.(AU)

Efeito do laser de baixa intensidade no procedimento de expansão rápida da maxila

A proposta do presente estudo foi avaliar os efeitos do laser de baixa intensidade na regeneração óssea no procedimento de expansão rápida da maxila. Utilizou-se 27 indivíduos com média de idade de 10,2 anos, divididos em dois grupos: grupo laser (n=14), no qual se realizou a expansão rápida da maxila, associada ao laser e grupo sem laser (n=13), que realizou somente a expansão rápida da maxila. O protocolo de ativação do parafuso expansor foi de 1 volta completa no primeiro dia e ½ volta diária até a sobrecorreção. O laser utilizado foi o de diodo (TWIN Laser MMOptics®, São Carlos), seguindo o protocolo de aplicação: comprimento de onda de 780nm, potência de 40mW, densidade de 10J/cm2, em 10 pontos localizados ao redor da sutura palatina mediana. Os estágios de aplicação foram: L1 (do primeiro ao quinto dia de aplicação), L2 (travamento do parafuso e 3 dias seguidos), L3, L4 e L5 (após 7, 14 e 21 dias do L2, respectivamente). Radiografias oclusais da maxila foram realizadas com auxílio de uma escala de alumínio, para referencial densitométrico, em diferentes tempos: T1 (inicial), T2 (dia de travamento do parafuso), T3 (3 a 5 dias do T2), T4 (30 dias do T3), T5 (60 dias do T4). As radiografias foram digitalizadas e submetidas a um programa de imagem (Image Tool - UTHSCSA, Texas, USA), para mensuração da densidade óptica das áreas previamente selecionadas. Para realização do teste estatístico, utilizou-se a Análise de Covariância usando como covariável o tempo para a fase avaliada. Em todos os testes foi adotado nível de significância de 5% (p<0,05).Para o Grupo Laser, os dados mostram que houve uma queda significante de densidade durante a abertura do parafuso (T2-T1), um aumento significante da mesma no período final de avaliação (T5-T4), e um aumento também da densidade no período de regeneração propriamente dito (T5-T2), ou seja, a partir do momento em que finalizou a fase de abertura do parafuso expansor. Enquanto que no Grupo Sem Laser, a densidade não mostrou diferença estatisticamente significantemente em nenhum período analisado. Os resultados mostraram que o laser propiciou consideravelmente uma melhor abertura da sutura palatina mediana, além de influenciar no processo de regeneração óssea da sutura, acelerando seus processos de reparo.(AU)

Further expansion of the alien seaweed Caulerpa taxifolia var. distichophylla (Sonder) Verlaque, Huisman & Procacini (Ulvophyceae, Bryopsidales) in the Eastern Mediterranean Sea

Date of Acceptance: 27/04/2015 We are grateful to Andreas Antoniou (Dep. of Environment, Ministry of Agriculture, Rural Development & Environment, Cyprus) for his assistance in the preparation of the illustrations. We would also like to thank Dr. Sotiris Orfanidis (NAGREF – Fisheries Research Institute, Kavala, Greece) for his valuable advice and both the DFMR and HSR / HCMR Rhodes crew and George Hatiris for their help in samplings. Special thanks are due to Dinos Leonidou (SeaQuest Divers Cyprus) for accompanying the deep dive for sampling Caulerpa at Cavo Greco. We are grateful to the Total Foundation (Paris) for its funding support to this study within the framework of the project “Brown algal ecology and biodiversity in the eastern Mediterranean Sea” and to the MASTS pooling initiative (Marine Alliance for Science and Technology for Scotland, funded by the Scottish Funding Council and contributing institutions; grant reference HR09011).

Experience-dependent, asymmetric expansion of hippocampal place fields

Theories of sequence learning based on temporally asymmetric, Hebbian long-term potentiation predict that during route learning the spatial firing distributions of hippocampal neurons should enlarge in a direction opposite to the animal’s movement. On a route AB, increased synaptic drive from cells representing A would cause cells representing B to fire earlier and more robustly. These effects appeared within a few laps in rats running on closed tracks. This provides indirect evidence for Hebbian synaptic plasticity and a functional explanation for why place cells become directionally selective during route following, namely, to preserve the synaptic asymmetry necessary to encode the sequence direction.

Synergies and trade-offs between renewable energy expansion and biodiversity conservation - a cross-national multifactor analysis

Peer reviewed

Expansion in vitro of transplantable human cord blood stem cells demonstrated using a quantitative assay of their lympho-myeloid repopulating activity in nonobese diabetic–scid/scid mice

Human hematopoiesis originates in a population of stem cells with transplantable lympho-myeloid reconstituting potential, but a method for quantitating such cells has not been available. We now describe a simple assay that meets this need. It is based on the ability of sublethally irradiated immunodeficient nonobese diabetic–scid/scid (NOD/SCID) mice to be engrafted by intravenously injected human hematopoietic cells and uses limiting dilution analysis to measure the frequency of human cells that produce both CD34−CD19+ (B-lymphoid) and CD34+ (myeloid) colony-forming cell progeny in the marrow of such recipients 6 to 8 weeks post-transplant. Human cord blood (CB) contains ≈5 of these competitive repopulating units (CRU) per ml that have a similar distribution between the CD38− and CD38+ subsets of CD34+ CB cells as long-term culture-initiating cells (LTC-IC) (4:1 vs. 2:1). Incubation of purified CD34+CD38− human CB cells in serum-free medium containing flt-3 ligand, Steel factor, interleukin 3, interleukin 6, and granulocyte colony-stimulating factor for 5–8 days resulted in a 100-fold expansion of colony-forming cells, a 4-fold expansion of LTC-IC, and a 2-fold (but significant, P < 0.02) increase in CRU. The culture-derived CRU, like the original CB CRU, generated pluripotent, erythroid, granulopoietic, megakaryopoietic, and pre-B cell progeny upon transplantation into NOD/SCID mice. These findings demonstrate an equivalent phenotypic heterogeneity amongst human CB cells detectable as CRU and LTC-IC. In addition, their similarly modest response to stimulation by a combination of cytokines that extensively amplify LTC-IC from normal adult marrow underscores the importance of ontogeny-dependent changes in human hematopoietic stem cell proliferation and self-renewal.

Expansion of a CUG trinucleotide repeat in the 3′ untranslated region of myotonic dystrophy protein kinase transcripts results in nuclear retention of transcripts

Expansion of a CTG trinucleotide repeat in the 3′ untranslated region (UTR) of DMPK, the gene encoding myotonic dystrophy protein kinase, induces the dominantly inherited neuromuscular disorder myotonic dystrophy (DM). Transcripts containing the expanded trinucleotide are abundant in differentiated cultured myoblasts, and they are spliced and polyadenylylated normally. However, mutant transcripts never reach the cytoplasm in these nonmitotic cells; instead, they form stable clusters that are tightly linked to the nuclear matrix, which can prevent effective biochemical purification of these transcripts. In DM patients, reduced DMPK protein levels, consequent to nuclear retention of mutant transcripts, are probably a cause of disease development. Formation of nuclear foci is a novel mechanism for preventing transcript export and effecting a loss of gene function.

Evolution of the Friedreich’s ataxia trinucleotide repeat expansion: Founder effect and premutations

Friedreich’s ataxia, the most frequent inherited ataxia, is caused, in the vast majority of cases, by large GAA repeat expansions in the first intron of the frataxin gene. The normal sequence corresponds to a moderately polymorphic trinucleotide repeat with bimodal size distribution. Small normal alleles have approximately eight to nine repeats whereas a more heterogeneous mode of large normal alleles ranges from 16 to 34 GAA. The latter class accounts for ≈17% of normal alleles. To identify the origin of the expansion mutation, we analyzed linkage disequilibrium between expansion mutations or normal alleles and a haplotype of five polymorphic markers within or close to the frataxin gene; 51% of the expansions were associated with a single haplotype, and the other expansions were associated with haplotypes that could be related to the major one by mutation at a polymorphic marker or by ancient recombination. Of interest, the major haplotype associated with expansion is also the major haplotype associated with the larger alleles in the normal size range and was almost never found associated with the smaller normal alleles. The results indicate that most if not all large normal alleles derive from a single founder chromosome and that they represent a reservoir for larger expansion events, possibly through “premutation” intermediates. Indeed, we found two such alleles (42 and 60 GAA) that underwent cataclysmic expansion to pathological range in a single generation. This stepwise evolution to large trinucleotide expansions already was suggested for myotonic dystrophy and fragile X syndrome and may relate to a common mutational mechanism, despite sequence motif differences.

Homeostatic expansion and phenotypic conversion of naïve T cells in response to self peptide/MHC ligands

Recent data suggest that survival of resting, naïve T cells requires an interaction with self MHC molecules. From analysis of the class I MHC-restricted T cell receptor transgenic strain OT-I, we report a different response. Rather than merely surviving, these T cells proliferated slowly after transfer into T-depleted syngeneic hosts. This expansion required both T cell “space” and expression of normal levels of self class I MHC molecules. Furthermore, we demonstrate that during homeostatic expansion in a suitable environment, naïve phenotype (CD44low) OT-I T cells converted to memory phenotype (CD44med/high), despite the absence of foreign antigenic stimulation. On the other hand, cells undergoing homeostatic expansion did not acquire cytolytic effector function. The significance of these data for reactivity of T cells with self peptide/MHC ligands and the implications for normal and abnormal T cell homeostasis are discussed.

A transient expansion of the native state precedes aggregation of recombinant human interferon-γ

Aggregation of proteins, even under conditions favoring the native state, is a ubiquitous problem in biotechnology and biomedical engineering. Providing a mechanistic basis for the pathways that lead to aggregation should allow development of rational approaches for its prevention. We have chosen recombinant human interferon-γ (rhIFN-γ) as a model protein for a mechanistic study of aggregation. In the presence of 0.9 M guanidinium hydrochloride, rhIFN-γ aggregates with first order kinetics, a process that is inhibited by addition of sucrose. We describe a pathway that accounts for both the observed first-order aggregation of rhIFN-γ and the effect of sucrose. In this pathway, aggregation proceeds through a transient expansion of the native state. Sucrose shifts the equilibrium within the ensemble of rhIFN-γ native conformations to favor the most compact native species over more expanded ones, thus stabilizing rhIFN-γ against aggregation. This phenomenon is attributed to the preferential exclusion of sucrose from the protein surface. In addition, kinetic analysis combined with solution thermodynamics shows that only a small (9%) expansion surface area is needed to form the transient native state that precedes aggregation. The approaches used here link thermodynamics and aggregation kinetics to provide a powerful tool for understanding both the pathway of protein aggregation and the rational use of excipients to inhibit the process.

Chimeric Cytokine Receptor Can Transduce Expansion Signals in Interleukin 6 Receptor α (IL-6Rα)-, IL-11Rα-, and gp130-low to -negative Primitive Hematopoietic Progenitors

We generated transgenic mice expressing chimeric receptors, which comprise extracellular domains of the human granulocyte–macrophage colony-stimulating factor (hGM-CSF) receptor and transmembrane and cytoplasmic domains of the mouse leukemia inhibitory factor receptor. In suspension cultures of lineage-negative (Lin−), 5-fluorouracil-resistant bone marrow cells of the transgenic mice, a combination of hGM-CSF and stem cell factor (SCF) induced exponential expansions of mixed colony-forming unit. The combination of hGM-CSF and SCF was effective on enriched, Lin−Sca-1+c-kit+ progenitors and increased either mixed colony-forming unit or cobblestone area–forming cells. In case of stimulation with hGM-CSF and SCF, interleukin-6 (IL-6) and SCF, or IL-11 and SCF, the most efficient expansion was achieved with hGM-CSF and SCF. When Lin−Sca-1+c-kit+CD34− further enriched progenitors were clone sorted and individually incubated in the presence of SCF, hGM-CSF stimulated a larger number of cells than did IL-6, IL-6 and soluble IL-6 receptor (IL-6R), or IL-11. These data suggest the presence of IL-6Rα-, IL-11Rα-, and gp130-low to -negative primitive hematopoietic progenitors. Such primitive progenitors are equipped with signal transduction molecules and can expand when these chimeric receptors are genetically introduced into the cells and stimulated with hGM-CSF in the presence of SCF.

Aggregation of huntingtin in yeast varies with the length of the polyglutamine expansion and the expression of chaperone proteins

Huntington's disease (HD) is an inherited neurodegenerative disorder caused by polyglutamine (polyQ) expansions in the huntingtin (Ht) protein. A hallmark of HD is the proteolytic production of an N-terminal fragment of Ht, containing the polyQ repeat, that forms aggregates in the nucleus and cytoplasm of affected neurons. Proteins with longer polyQ repeats aggregate more rapidly and cause disease at an earlier age, but the mechanism of aggregation and its relationship to disease remain unclear. To provide a new, genetically tractable model system for the study of Ht, we engineered yeast cells to express an N-terminal fragment of Ht with different polyQ repeat lengths of 25, 47, 72, or 103 residues, fused to green fluorescent protein. The extent of aggregation varied with the length of the polyQ repeat: at the two extremes, most HtQ103 protein coalesced into a single large cytoplasmic aggregate, whereas HtQ25 exhibited no sign of aggregation. Mutations that inhibit the ubiquitin/proteasome pathway at three different steps had no effect on the aggregation of Ht fragments in yeast, suggesting that the ubiquitination of Ht previously noted in mammalian cells may not inherently be required for polyQ length-dependent aggregation. Changing the expression levels of a wide variety of chaperone proteins in yeast neither increased nor decreased Ht aggregation. However, Sis1, Hsp70, and Hsp104 overexpression modulated aggregation of HtQ72 and HtQ103 fragments. More dramatically, the deletion of Hsp104 virtually eliminated it. These observations establish yeast as a system for studying the causes and consequences of polyQ-dependent Ht aggregation.

Regulated expression of P210 Bcr-Abl during embryonic stem cell differentiation stimulates multipotential progenitor expansion and myeloid cell fate

P210 Bcr-Abl is an activated tyrosine kinase oncogene encoded by the Philadelphia chromosome associated with human chronic myelogenous leukemia (CML). The disease represents a clonal disorder arising in the pluripotent hematopoietic stem cell. During the chronic phase, patients present with a dramatic expansion of myeloid cells and a mild anemia. Retroviral gene transfer and transgenic expression in rodents have demonstrated the ability of Bcr-Abl to induce various types of leukemia. However, study of human CML or rodent models has not determined the direct and immediate effects of Bcr-Abl on hematopoietic cells from those requiring secondary genetic or epigenetic changes selected during the pathogenic process. We utilized tetracycline-regulated expression of Bcr-Abl from a promoter engineered for robust expression in primitive stem cells through multilineage blood cell development in combination with the in vitro differentiation of embryonal stem cells into hematopoietic elements. Our results demonstrate that Bcr-Abl expression alone is sufficient to increase the number of multipotent and myeloid lineage committed progenitors in a dose-dependent manner while suppressing the development of committed erythroid progenitors. These effects are reversible upon extinguishing Bcr-Abl expression. These findings are consistent with Bcr-Abl being the sole genetic change needed for the establishment of the chronic phase of CML and provide a powerful system for the analysis of any genetic change that alters cell growth and lineage choices of the hematopoietic stem cell.