Estudos epidemiológicos regionais que envolvam a traumatologia maxilofacial são importantes para auxiliar o desenvolvimento de formas mais eficientes de prestar cuidados em saúde, para avaliar e melhorar a qualidade destes cuidados e gerenciar estratégias de prevenção, financiamento e acesso ao tratamento no setor público de saúde. O tempo decorrido entre um trauma de face de um paciente até o atendimento clínico e tratamento das fraturas pode afetar os resultados clínicos em termos de intervenções resolução e complicações. O objetivo deste estudo foi verificar se no Hospital Público Eugênio Espejo de Quito, existe uma relação entre variáveis de tipificação das fraturas, seus tratamentos e os intervalos de tempo para sua resolução clínica com o aparecimento de complicações. Um estudo retrospectivo foi realizado nos prontuários do Serviço de Cirurgia bucomaxilofacial veificando dados epidemiológicos e da evolução clínica de pacientes que sofreram fraturas em face, esqueleto fixo e mandíbula entre os anos de 2012 e 2016. Os resultados mostraram que a epidemiologia das fraturas faciais foi bastante semelhante ao que a literatura pertinente ao assunto tem mostrado. A maioria dos casos ocorreram fora da cidade de Quito (64%). Observamos que não existe associação das variáveis: histórico de hábitos nocivos, presença de desvio, tipo de fratura, abordagem da fratura, tipo de fixação, presença ou não de comorbidades com o aparecimento de fraturas. Entretanto, os dados mostraram que existe 70% mais chances de risco de complicações quando o intervalo decorrido entre o acidente e o tratamento definitivo ultrapassar uma semana. Discutimos os fatores que podem implicar neste maior intervalo de tempo entre essas condutas e concluímos que existe a necessidade de investimento na especialidade e estudos para implantação de novos serviços fora da cidade de Quito. / Regional epidemiological studies involving maxillofacial traumatology are important to assist in the development of more efficient ways of providing health care, to assess and improve the quality of these care and to manage strategies of prevention, funding and access to treatment in public health sector. The time elapsed between a trauma of a patient\'s face until clinical care and treatment of fractures can affect the clinical results in terms of resolution interventions and complications. The objective of this study was to check whether in the public hospital Eugenio Espejo de Quito, there is a relationship between variables of typing of the fractures, their treatments and the intervals of time for their clinical resolution with the appearance of complications. A retrospective study was performed in the records of the Maxillofacial Surgery Service to verify epidemiological data and clinical evolution of patients who suffered fractures in face, fixed skeleton and jaw, between 2012 and 2016. The results showed that epidemiology of the facial fractures was quite similar to what pertinent literature to the subject has shown. Many of the cases occurred out city of Quito (64%). We observe that there is no association of historical variables of harmful habits, presence of deviation, type of fracture, approach of fracture, type of fixation, presence or not of comorbidities with the appearance of complications. However, the data showed that there are 70% more chances of risk of complications when the interval elapsed between the accident and the definitive treatment exceed one week. We discuss the factors that can be associated with the largest interval of time between conducts and we conclude that there is a need for investment in the specialty and studies for implementation of new services out the city of Quito.

O feijão caupi, através da simbiose com bactérias do gênero Bradyrhizobium, pode obter nitrogênio através do processo de fixação biológica do N2 (FBN). A nodulação e a FBN são influenciadas por fatores edafoclimáticos que podem trazer benefícios ou prejuízos ao processo. A disponibilidade de nutrientes está entre os principais fatores que influenciam a FBN e, dentre os principais nutrientes que influenciam tal processo, cita-se o fósforo.Objetivou-se com esse trabalho, avaliar o rendimento de cultivares de feijão caupi em função das estirpes fixadora de nitrogênio associada ou não a adubação fosfatada, no sul do estado do Tocantins em condições de casa de vegetação e em campo. Os experimentos foram conduzidos em casa de vegetação e a campo, um com a cultivar de feijão-caupi, BRS Nova Era e o outro BRS Sempre Verde, com 4 estirpes de rizóbio mais um tratamento sem bactéria, associadas ou não a 6 doses de fósforo.O delineamento utilizado foi em blocos casualizados, disposto em esquema fatorial 5 x 6. A campo os tratamentos foram dispostos em um delineamento experimental blocos casualizados sob arranjo fatorial 3 x 6, sendo o primeiro fator duas estirpes de rizóbio e um tratamento sem e o segundo fator seis doses de P2O5 com 4 repetições. As doses consistiram de 0, 30, 60, 90, 120 e 150 kg ha-1de P2O5, aplicadas na semeadura. As características avaliadas em casa de vegetação foram: altura de planta, diâmetro de colo, massa seca da parte aérea, massa seca das raízes, número de nódulos, massa seca dos nódulos. As características avaliadas em campo foram: clorofila total, massa seca da parte aérea, peso de cem sementes, produtividade e nitrogênio total na parte aérea. As estirpes estudadas associadas ou não as doses de fósforo proporcionaram acréscimo na altura de planta, diâmetro de colmo, massa seca da parte aérea, massa seca da raiz para as duas cultivares de feijão-caupi estudas BRS Nova Era e BRS Sempre Verde. Para os experimentos de campo a cv. BRS Sempre Verde a estirpe mais indicada é INPA 03-11B; para a cv. BRS Vinagre a estirpe BR 3299 foi mais eficiente. A dose de fósforo que condicionou a máxima eficiência agronômica variou de 70 a 110 kg ha-1. / Cowpea beans, through symbiosis with bacteria of the genus Bradyrhizobium, can obtain nitrogen through the biological fixation process of N2 (BNF). Nodulation and BNF are influenced by edaphoclimatic factors that can bring benefits or losses to the process. The availability of nutrients is among the main factors influencing BNF and, among the main nutrients that influence this process, phosphorus is cited. The objective of this work was to evaluate the yield of cowpea cultivars as a function of nitrogen fixing strains associated or not with phosphate fertilization, in the southern state of Tocantins under greenhouse conditions and in the field. The experiments were conducted in a greenhouse and in the field, one with the cowpea bean cultivar BRS Nova Era and the other BRS Sempre Verde with four strains of rhizobium plus a treatment without bacteria, associated or not To 6 doses of phosphorus. The experimental design was a randomized block design, arranged in a factorial scheme 5 x 6. In the field treatments were arranged in a randomized block design under a factorial arrangement 3 x 6, the first factor being two strains of rhizobia and A treatment without and the second factor six doses of P2O5 with 4 replicates. The doses consisted of 0, 30, 60, 90, 120 and 150 kg ha-1 of P2O5, applied at sowing. The characteristics evaluated in greenhouse were plant height, diameter of lap, dry mass of shoot, dry mass of roots, number of nodules, dry mass of nodules. The characteristics evaluated in the field were: total chlorophyll, shoot dry mass, weight of one hundred seeds, productivity and total nitrogen in shoot. The strains studied, whether or not associated with phosphorus doses, increased plant height, stem diameter, dry shoot mass, root dry mass for the two bean cultivars BRS Nova Era and BRS Sempre Verde. For the field-to-cv. BRS sempre verde the most indicated strain is INPA 03-11B; For cv. BRS Vinagre strain BR 3299 was more efficient. The dose of phosphorus that conditioned the maximum agronomic efficiency ranged from 70 to 110 kg ha-1.

Introdução: Os pacientes com fissura labiopalatina, com deficiências maxilares muito severas, geralmente são tratados com avanço maxilar por meio da osteotomia tipo Le Fort I. Entretanto, a distração osteogênica com o distrator externo rígido (RED) pode funcionar como uma alternativa terapêutica para a correção da discrepância esquelética. Proposição: O objetivo do presente estudo é avaliar os efeitos do avanço maxilar por meio da distração osteogênica com distrator externo rígido (RED), associada à osteotomia tipo Le Fort I, em pacientes com fissura transforame unilateral ou bilateral, quanto à quantidade de avanço maxilar e à sua estabilidade a médio e longo prazo. Materiais e Métodos: Para a realização deste estudo longitudinal e retrospectivo, foram usadas telerradiografias em norma lateral de 9 pacientes (6 do gênero masculino e 3 do gênero feminino), onde 4 apresentaram fissura transforame unilateral e 5 apresentaram fissura transforame bilateral, submetidos ao avanço maxilar por meio da distração osteogênica com distrator externo rígido (RED). Foram estabelecidos três tempos de avaliação: fase pré-distração (T1), fase pós-distração imediata (T2) e fase pós-distração controle, com o mínimo de 1 ano após a finalização da distração (T3). A demarcação dos pontos cefalométricos e a obtenção das medidas das variáveis cefalométricas foram realizadas através do software Dolphin Imaging®, versão 11.5. Para a análise dos resultados, o teste estatístico ANOVA de medidas repetidas foi utilizado, adotando-se o nível de significância de 5%. Resultados: No início da distração, a idade média foi de 14 anos e 4 meses (idade mínima de 9 anos, e máxima de 21 anos). O período médio de distração foi de 18 dias, com uma média de ativação no distrator de 1,0mm/dia. O avanço médio da maxila medido em LVR-A, em T2, foi de 15,6mm (p<0,001), com recidiva não estatisticamente significante de 21,79% (p=0,102), em T3. O aumento médio de SNA, em T2, foi de 14,8º (p<0,001), com recidiva não estatisticamente significante de 18,90% (p=0,130), em T3. Os valores médios das medidas SN.GoMe, 1.PP e IMPA não apresentaram variação estatisticamente significante (p>0,05) entre T1, T2 e T3. Conclusão: A terapia de distração osteogênica para avanço maxilar com o RED mostrou ser eficiente, com aumentos significantes das medidas cefalométricas lineares e angulares relacionadas ao avanço maxilar, demonstrando efeito predominantemente esquelético, e estabilidade no período pós-distração médio (T3) de 1 ano e 8 meses. / Introduction: Patients with unilateral and bilateral cleft lip and palate, with significant maxillary hypoplasia are commonly treated with maxillary advancement by Le Fort I osteotomy. However, distraction osteogenesis with a rigid external distraction (RED) device can function as an alternative option for treatment of the skeletal discrepancy. Purpose: The aim of this study is to assess the effects of maxillary advancement by distraction osteogenesis using a rigid external distraction (RED) device, associated with the Le Fort I osteotomy in patients with unilateral or bilateral cleft lip and palate, as the amount of maxillary advancement and their stability in the medium and long term. Materials and Methods: To perform this retrospective longitudinal study, lateral cephalograms of 9 patients (6 males and 3 females) were used, where 4 had unilateral cleft lip and palate and 5 had bilateral cleft lip and palate, who underwent maxilla advancement by distraction osteogenesis with RED device. Three stages of evaluation were established: pre-distraction (T1), immediate post-distraction (T2) post-distraction control, with a minimum of 1 year after completion of distraction (T3). The anatomic landmarks and measurements of cephalometric variables were performed by using the Dolphin Imaging® version 11.5 software. To evaluate the results, the ANOVA test for repeated measures was used, adopting a significance level of 5%. Results: At the start of distraction, mean age was 14 years and 4 months (minimum age 9 years old and maximum of 21 years old). The mean distraction period was 18 days, with a mean rate of distractor activation in 1.0 mm / day. The mean maxillary advancement in LVR-A, at T2, was 15.6 mm (p<0.001), with no statistically significant relapse of 21.79% (p=0.102) at T3. The SNA angle increase, at T2, was 14.8º (p<0.001), with no statistically significant relapse of 18.90% (p=0.130), at T3. The mean values of SN.GoMe, IMPA and 1.PP measures showed no statistically significant variation (p>0.05) between T1, T2 and T3. Conclusion: The therapy of distraction osteogenesis for maxillary advancement with RED is efficient, with significant increases in the linear and angular cephalometric measurements related to the maxilla advancement, demonstrating predominantly skeletal effect and stability in mean post-distraction period (T3) of 1 year and 8 months.

Enterobactérias são comumente encontradas em simbiose com plantas, promovendo o crescimento vegetal, através da redução do nitrogênio atmosférico a amônia. Entretanto, são os principais patógenos na medicina humana e veterinária. O objetivo deste estudo foi buscar marcadores genéticos e fisiológicos que caracterizem espécies de enterobactérias isoladas de cana-de-açúcar (n=24) e de origem clínica (n=15). Foram submetidas a testes moleculares e fisiológicos como: capacidade de reduzir N2; perfil de susceptibilidade a antibióticos; atividade hemolítica; produção de substâncias promotoras de crescimento e liberação de enzimas extracelulares. Atividades metabólicas e fisiológicas altamente conservadas, sustentadas pelo ponto vista genético, permitem a distribuição destas espécies em diferentes ecossistemas, e, em condições favoráveis, esta adaptação pode contribuir para o estabelecimento de processos patogênicos ou simbióticos em plantas. Então, enterobacterias ambientais podem se tornar patogênicas, enquanto isolados clínicos podem se adaptar a condições ambientais. / Enterobacteria are commonly found in symbiosis with plants, promoting their growth by reducing atmospheric nitrogen to ammonia. However, they are also major pathogens in human and veterinary medicine. The aim of this study was to characterize physiological and genetic markers that distinguish enterobacteria species isolated from sugarcane (n= 24) and clinical sources (n= 15). The samples were characterized regarding their molecular and physiological assays as: ability to reduce N2, antibiotic susceptibility profile; screen for hemolytic activity; production of plant-growth promoting compounds and release of extracellular enzymes. Finally, highly conserved physiological and metabolic activities are supported on genetic backgrounds that allow a wide distribution along different ecosystems and, in favorable conditions, this adaptation could contribute to the establishment of a pathogenic process or symbiosis in plants. So, environmental enterobacteria isolates can become pathogenic, whereas clinical strains can adapt to environmental conditions.

Raramente plantações de eucalipto respondem à adubação nitrogenada. Contudo, por se tratar de povoamentos monoespecíficos, com baixa capacidade de fixação de N, depois de algumas rotações de cultivo a probabilidade de resposta a este nutriente aumenta, devido ao esgotamento das reservas naturais de nitrogênio do solo acumuladas no ecossistema anterior. O presente estudo teve como objetivos avaliar i) o crescimento, o desenvolvimento de raízes finas de Eucalyptus grandis e de Acacia mangium em povoamentos monoespecíficos e mistos; ii) a nodulação da A. mangium nestes povoamentos. As árvores foram plantadas em maio de 2003 no espaçamento 3m x 3m. O delineamento experimental foi o de blocos completamente aleatorizados (4 blocos), sendo avaliados 3 tratamentos: povoamentos monoespecíficos de E. grandis e A. mangium e um povoamento em que essas espécies foram consorciadas (1100 plantas por ha de E. grandis e 550 árvores por ha de A. mangium). Quando consorciada, a A. mangium foi plantada no meio da distância entre as árvores de E. grandis na mesma linha de plantio. Foram amostradas raízes finas vivas (&#8804;3 mm) em 5-16 posições (dependendo do tratamento), de uma árvore de área basal média em doze parcelas (3 tratamentos x 4 blocos), até 1 m de profundidade aos 6 e 12 meses pós-plantio e até 2 m de profundidade aos 18 e 30 meses pós-plantio, por meio de uma sonda de aço com 4,5 cm de diâmetro interno. O E. grandis foi diferente da A. mangium quanto ao desenvolvimento radicular nos diferentes períodos de amostragem, principalmente na camada superficial do solo (0-10 cm). No povoamento misto, aos 30 meses pós-plantio, o E. grandis foi 64% maior em altura do que a A. mangium e 75% maior em área basal, produzindo 94% mais biomassa do que esta leguminosa. A assimetria do crescimento radicular foi elevada entre as espécies, reflexo da grande competição por luz na parte aérea. Não houve clara estratificação de exploração das camadas do solo pelo sistema radicular das espécies. O E. grandis apresentou maior exploração radicular em todas as camadas do solo, fazendo com que o desenvolvimento radicular da A. mangium ficasse restrito a proximidade da árvore. O elevado grau de assimetria no desenvolvimento radicular entre as espécies resultou, aos 30 meses, na camada 0-10 cm, numa diferença de 91% na densidade de raízes finas (drf) entre as espécies. A nodulação foi sensível a competição com o E. grandis, aparentemente, devido à variação de umidade e temperatura sazonal. Aos 18 meses, no povoamento monoespecífico, houve tendência de maior densidade de nódulos na linha de plantio, camada 0-10 cm. Aos 30 meses os nódulos foram encontrados próximos às árvores. No misto, aos 18 e 30 meses ocorreu diminuição da densidade de nódulos na camada 10-30 cm e aumento entre 30 e 50 cm. / Eucalypts plantations rarely answer to the nitrogen fertlization. However, for being of monospecific stands, with low capacity of N fixation, after some cultivation rotations the answer probability to this nutritious one increases, due to the exhaustion of the natural reservations nitrogen of the soil accumulated in previous ecosystem. The present study had as objectives to evaluate i) the growth, the development of Eucalyptus grandis and Acacia mangium fine-root in monospecific and mixed stands; ii) the A. mangium nodulation in these stands. The trees were planted in may of 2003 in the spacing 3m x 3m. The experimental design was it of completely randomized blocks (4 blocks), between assessed 3 treatments: Monospecific stands of E. grandis and A. mangium and a stand in that those species were mixed (1100 plants for ha of E. grandis and 550 trees for ha of A. mangium). When mixed, to A. mangium was planted in the middle of the distance among the trees of E. grandis in the same plantation line. They were sampling live fine-root (&#8804; 3 mm) in 5-16 positions (depending on the treatment), of a tree of medium basal area in twelve portions (3 treatments x 4 blocks), until 1 m of depth to the 6 and 12 months powder-plantation and until 2 m of depth to the 18 and 30 months powder-plantation, by means of a sledgehammer with 4,5 cm of internal diameter. The E. grandis was different from the A. magium with relationship to the roots development in the different sampling periods, mainly in the superficial layer of the soil (0-10 cm). In the mixed stand, between 29 and 30 months after plantation, E. grandis was 64% larger in height than at A. mangium and 75% larger in basal area, producing 94% more biomass than this leguminous. The asymmetry of the root growth was elevated between the species, reflex of the great competition for light in the aerial part. There was not clear stratification of exploration of the layers of the soil for the roots system of the species. The E. grandis presented larger roots exploration radicular in all the layers of the soil, doing with that the roots development of the A. mangium was restricted in the proximity of the tree. The high asymmetry degree in the roots development between the species, resulted to the 30 months in the layer 0-10 cm, a difference of 91% in the density of fine roots (dfr) between the species. The nodulation was sensitive the competition with the E. grandis , seemingly, due to the humidity variation and seasonal temperature. To the 18 months, in the monospecific stands, there was tendency of larger density of nodules in the plantation line, layer 0-10 cm. To the 30 months the nodules were found close to the trees. In the mixed, to the 18 and 30 months it happened decrease of the density of nodules in the layer 10-30 cm and increase between 30 and 50 cm.

Com o objetivo de se estudar a estabilidade da resistência mecânica da fixação pedicular do segmento vertebral, foram realizados ensaios mecânicos de sistemas de fixação pedicular compostos por parafusos hastes, sapatas e porcas, fabricados em titânio. Os ensaios foram flexo-compressão e flexão lateral, realizados na máquina universal de ensaio com a finalidade de se avaliar a influência da estrutura dos parafusos pediculares canulados (ocos) e não canulados (maciços) e a utilização ou não das sapatas na estabilidade de fixação do sistema. Os ensaios foram realizados na fase elástica, durante todo o tempo de aplicação das cargas. Foram realizadas três combinações de cada ensaio, com troca dos parafusos e sapatas do seguinte modo: parafusos canulados com sapata (CCS), canulados sem sapata (CSS) e não canulados sem sapata (NCSS). Os corpos de prova foram confeccionados em madeira mogno, e foram ensaiadas dez montagens de cada combinação, para os dois tipos de ensaios. Os resultados dos ensaios permitiram o cálculo do momento fletor para os valores das deflexões máximas em cada ensaio, assim como a rigidez e a energia absorvida. No ensaio de flexo-compressão, a análise de variância do momento fletor entre os grupos mostrou que não houve diferença significativa (CCS x CSS (p=0,304; CCS x NCSS (p=0,447); CSS x NCSS (p=0,183). O mesmo resultado foi encontrado para a rigidez (CCS x CSS (p=0,988); CCS x NCSS (p=0,500); CSS x NCSS (p=0,272), e para a energia absorvida (CCS x CSS (p=0,751); CCS x NCSS (p=0,673); CSS x NCSS (p=0,738). No ensaio de flexão lateral, a análise de variância do momento fletor entre os grupos mostrou que não houve diferença significativa (CCS x CSS (p=0.304); CCS x NCS (p=0,883); CSS x NCSS (p=0,186). O mesmo resultado foi encontrado para a rigidez (CCS x CSS (p=0,285); CCS x NCSS (p=0,957); CSS x NCSS (p=0,147), e para a energia absorvida (CCS x CSS (p=0,304); CCS x NCSS (p=0.892): CSS x NCSS (p=0,182). / Mechanical tests of the pedicle fixation systems composed of screws, rods, devices and nuts, made of titanium were realised with the purpose of studying the stability of mechanical resistance of the pedicle fixation of vertebral segments. The tests were bending-compression and lateral flexion done in the universal machine of test aiming at analysing the influence oft he structure of the hollowed pedicles screws and massy pedicles screws and the use or not of the devices in the stability of system fixation. The tests were carried out during the elastic phase, while the loads were being applied. Three combinations of each test were carried out, exchanging the devices and screws in the following way: screws with a device (CCS), screws without a device (CSS) and massy screws without devices (NCSS). The implants were made of \"mogno\" and ten (10) specimens used for each combination for both kinds of tests. The results of the tests allowed the calculation of the bending moment for the value in the maximum deflection of each test as well as the stiffness and energy absorbed. In test of bending compression, the analysis of variation of the bending moment among the groups showed that there was no difference (CCS x CSS (p=0,304); CCS x NCSS (p=0,447); CSS x NCSS (p=0,183)). The same result was found for the stiffness (CCS x CSS (p=0,988); CCS x NCSS (p=0,500); CSS x NCSS (p=0,272)), and the absorbed energy (CCS x CSS (p=0,751); CCS x NCSS (p=0,673); CSS x NCSS (p=0,738)). In the flexion lateral, the analysis of the variation of the bending moment the groups showed no difference (CCS x CSS (p=0,304); CCS x NCSS (p=0,883); CSS x NCSS (p=0,186)). The same result was found for the stiffness (CCS x CSS (p=0,285); CCS x NCSS (p=0,957); CSS x NCSS (p=0,147), and the absorbed energy (CCS x CSS (p=0,304); CCS x NCSS (p=0,892); CSS x NCSS (p=0,182)).

This diploma thesis deals with common parasites of game animals, animals and human occurring in the territory of the Czech Republic. It focuses on their fixation and general preparation of such specimens for usage in teaching of Science. The first part is devoted to the phenomenon of parasitism, and the history and definition of this concept. Systematic hierarchy of various unrelated parasitic groups is clarified. The second part provides the list of parasites, from the evolutionary original to the highly derived groups. We give an overview of their morphology and anatomy with the emphasis on their life cycle. The detailed knowledge of life cycle is essential for obtaining specimens. In each group, we indicate methods of collection and fixation in detail. The co called smartmicroscope represent the main point of the third part of proposed diploma thesis. Text of the thesis is supplemented with pictures taken by smart microscope, as well as by common microscope. KEY WORDS parasite, parasitism, mikropredatore, vector, fixation, preparation, smart mikroscope

临床及动物实验的文献报告表明, 低能量脉冲超声波 (LIPUS) 能促进骨折愈合。 可是, 不同研究小组针对LIPUS的功效所提供的数据结果往往并不一致。为了找出导致数据结果不一致的原因, 以及提升LIPUS的生物功效, 科研人员正致力于测定超声波在骨折治疗中的最佳信号参数。 在临床运用上, LIPUS对骨折的治疗一般是以经皮方式应用的。 故此, 不同层次深度的骨折会暴露在不同的超声波场区里。 超声波场有两个不同的区域, 就是近场区 (接近超声波换能器的区域) 及远场区 (远离超声波换能器的区域)。 在我们早期的临床研究中, 我们曾使用超声波的近场区治疗胫骨的复杂性骨折。 我们发现, 当超声波换能器置于胫骨骨折处的前方, 骨痂集中生成于胫骨骨折处的背面。 这研究结果显示, 近场区以外的超声波场或许更具促进骨痂形成的功效。 再者, 针对LIPUS声场仿真分析的结果显示, 近场区内的声压分布是不稳定的, 而远场区内的声压则远比近场区的均及稳定。由于声压的稳定性会大大影响超声波于组织内的能量透射, 我们相信在超声波场中, 骨折的深度会影响LIPUS的生物效应。 / 本研究采用大鼠闭合性股骨骨折模型及细胞培养实验, 探究不同的超声波场区对骨折愈合的影响。本研究作出了以下三个科研假设: (1) LIPUS 的远场区在促进骨折愈合的应用上有着更高的生物效应; (2) LIPUS 的远场区能透过促使骨细胞产生旁分泌调节因子, 从而提升成骨细胞的成骨活性; (3) 通过换能器直径的调制而产生的LIPUS远场区能有效地促进骨折愈合。 / 在第一部分的实验里, 股骨骨折的SD大鼠被随机分为对照组 (control), 近场区超声波治疗组 (near field; 伤肢跟换能器相距0mm), 中近场区超声波治疗组 (mid-near field; 伤肢跟换能器相距60mm), 远场区超声波治疗组 (far field; 伤肢跟换能器相距130mm)。在伤肢及超声波换能器之间安放了跟软组织具有同一超声波衰减系数的凝胶 (长度: 0mm, 60mm, 130mm)。LIPUS每天治疗20分钟, 每周治疗5天。 我们研究结果显示, 治疗后的第四周, 远场治疗组的骨痂组织具有各组中最高的相对骨体积及组织矿密度, 这造就远场治疗组相比对照组具有更强的力学属性。我们的研究结果表明, LIPUS的远场区治疗能通过提升骨痂的骨体积及骨矿化, 进一步促进骨折的愈合。 / 在第二部分的实验里, 我们把骨细胞株(MLO-Y4) 暴露在三種不同的超声波場中: 0 mm, 60 mm 及130 mm 。 经过不同的LIPUS处理后, 我们把条件培养基(CM) 收集, 并将其用于培养成骨前趋细胞株(MC3T3-E1)。 这部分的实验共有5组: Non组(非条件培养基处理组), Con组(骨细胞条件培养基处理组), 0mm组(条件培养基处理组; 条件培养基收集自经过LIPUS近场区刺激后的骨细胞), 60mm组(条件培养基处理组; LIPUS中近场区刺激后的骨细胞), 以及 130mm组(条件培养基处理组; LIPUS远场区刺激后的骨细胞)。我们测试了各超声波场对骨细胞的直接影响, 以及成骨前趋细胞经过各类骨细胞条件培养基培养后的成骨活性。 免疫染色的结果显示近场区以后的超声波场 (130mm 及 60mm) 能进一步诱导β-catenin 于骨细胞的入核作用。 另外, 远场区的骨细胞条件培养基 (130mm CM) 的处理促进了成骨前趋细胞的: (1) 细胞迁移的能力 (反映自细胞伤口愈合测试) ; (2) 细胞分化成熟的机制 (BrdU细胞增殖检验及ALP活性分析): (3) 基质钙化 (Alizarin red 钙化结节染色)。 / 在第三部分的实验里, 我们把换能器的直径缩减致一半, 以致换能器跟LIPUS远场区之间的距离从130 mm被拉近至30 mm。 当LIPUS以经皮的方式应用, 位于皮下大约40 mm的大鼠股骨骨折因此暴露在LIPUS的远场区。 相对于在换能器及伤肢之间安置130 mm凝胶, 以调制换能器直径而直接让骨折暴露于LIPUS远场区是更具临床应用性的方法。 在这部分, 股骨骨折的SD大鼠被随机分为对照组 (control), LT-Near30 (正常的换能器直径; 近场超声处理; I[subscript SATA] = 30 mW/cm²), ST-Far30 (缩减后的换能器直径; 远场超声处理; I[subscript SATA] = 30 mW/cm²), ST-Far150 (缩减后的换能器直径; 远场超声处理; I[subscript SATA] = 150 mW/cm²)。 研究结果证实, 以调制换能器直径而产生的远场LIPUS (ST-Far30)能透过提升骨痂的生成及力学属性, 进一步促进骨折愈合。 同时, 我们的结果显示, 相对高强度 (150 mW/cm²) 的远场LIPUS治疗不能更有效地促进骨折愈合。 / 综上所述, 动物及细胞培养实验结果证明, LIPUS的远场区在促进骨折愈合上更具功效。由于LIPUS的远场区放射稳定的超声波束, 骨痂中的骨细胞受引发释放可溶性因子, 从而进一步激发成骨样细胞的成骨活性。 这些细胞的生物效应造就LIPUS的远场区在促进骨折愈合上更具治疗效果。最后, 我们亦把以上的研究发现转化成具临床应用性的LIPUS应用方法。 这应用方法能让超声波换能器以紧贴皮肤的方式直接使骨折暴露于LIPUS远场区, 从而达成促进骨折愈合的功效。 / Low-intensity Pulsed Ultrasound (LIPUS) has been confirmed to enhance fracture healing in many clinical and animal studies. However, the evidences from literatures to support the applications of LIPUS on fracture healing were inconsistent. Therefore, scientists have been studying various ultrasound parameters aiming to find out the factors resulting in the inconsistent outcomes among research groups, and to further enhance the efficacy of LIPUS. Clinically, LIPUS is usually applied onto fracture sites transcutaneously, hence, fractures at different depths are exposed to different zones of ultrasound beam. There are two characteristic zones of ultrasound beam: the near field (close to the transducer) and far field (farther from the transducer). In our previous clinical study, direct transcutaneously applied LIPUS (near field LIPUS exposure) was used to treat human tibial complex fractures. We found that callus usually formed on the posterior side when the transducer was placed on the anterior side. This finding implied that ultrasound beam beyond near field bears higher potential in promoting callus formation. Moreover, beam mapping measurement of LIPUS shows a variable spatial pressure in near field; while a more uniform pressure profile was found beyond it (far field). As the stability of pressure profile influences the ultrasound energy transmission in tissue, we postulate that the biological effects of LIPUS are affected by the fracture depths within the ultrasound field. / This study aims to address the research question of how ultrasound fields influence the fracture healing through testifying the following hypotheses in animal and cell culture studies: (1) Far field LIPUS bears higher biological effect in facilitating fracture healing; (2) Far field LIPUS could enhance the osteogenic activities of osteoblastic cells via paracrine factors secreted from osteocytes; (3) Far field LIPUS setup by transducer diameter modulation could facilitate fracture healing. / In part one study, femoral fractured Sprague-Dawley (SD) rats were randomized into control, near-field (fractures placed at 0mm away from transducer), mid-near field (60mm away from transducer) or far-field (130mm away from transducer) groups. Rubber gel block (lengths: 0mm, 60mm and 130mm) with attenuation coefficient equivalent to soft tissue was interposed between the transducer and the fractured limb. LIPUS was given 20min/day and 5days/week. We found the callus in 130mm group was the highest in bone volume fraction and tissue mineral density at week 4. These advancements mutually contributed to its significantly stronger mechanical properties than the control group. Our results indicated that far field LIPUS could further facilitate fracture healing by promoting bone volume increase and callus mineralization, which led to enhanced mechanical properties. / In part two study, LIPUS was applied to osteocyte cell line (MLO-Y4) at three distances: 0mm, 60mm and 130mm. The conditioned medium (CM) collected from different LIPUS treatment regimens were used to culture pre-osteoblast cell line (MC3T3-E1). There were 5 groups in the CM treatment: Non group (plain α-MEM treatment), Con group (osteocyte CM), 0mm group (Near field LIPUS treated osteocyte CM), 60mm group (Mid-near field LIPUS treated osteocyte CM) and 130mm group (Far field LIPUS treated osteocyte CM). The effect of ultrasound fields on osteocytes and the osteogenic activities of the pre-osteoblasts after different CM treatments were assessed. The immunostaining results indicated that beyond near field LIPUS (LIPUS at 130mm and 60mm) could further promote β-catenin nuclear translocation in osteocytes. The far field LIPUS osteocyte-CM (130mm group) caused the highest biological effect on (1) pre-osteoblasts migration (reflected by wound healing assay); (2) maturation of pre-osteoblasts: transition of cell proliferation into osteogenic differentiation (BrdU cell proliferation assay and ALP activity assay); and (3) matrix calcification (Alizarin red calcium nodule staining). / In part three study, the transducer diameter was reduced by half in order to draw the far field location closer to the transducer (i.e. from 130 mm to 30 mm). As the femoral shaft fractures of rats are located at around 40 mm beneath the skin, fractures were directly exposed to far field LIPUS transcutaneoulsy. It is a more clinically applicable approach than the method of physical separation (i.e. 130 mm separation between transducer and fractured limb). Femoral fractured SD rats were randomized into control, LT-Near30 (conventional transducer diameter, near field, I[subscript SATA] = 30 mW/cm²), ST-Far30 (small transducer, far field, I[subscript SATA] = 30 mW/cm²) and ST-Far150 (small transducer, far field, I[subscript SATA]=150 mW/cm²). Our results confirmed that the far field LIPUS emitted from the transducer diameter reduction setup (ST-Far30) could further facilitate fracture healing process by enhancing callus formation and mechanical properties. Our findings also indicated that fractures exposed to far field LIPUS with relatively higher intensity (150 mW/cm²) did not heal better. / In summary, our in vivo and in vitro findings reinforce each other to confirm the positive effects of far field LIPUS on promoting fracture healing. As far field LIPUS radiates a stable ultrasound beam, osteocytes inside the callus are triggered to secrete soluble factors to promote the osteogenic activities of osteoblastic cells. This contributes to the higher therapeutic effects of far field LIPUS on fracture healing. We also translated these findings to establish a clinically applicable LIPUS device, which directly radiates far field LIPUS to subcutaneous fracture site without any distance separation needed. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Fung, Chak Hei. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 186-207). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Abstract --- p.i / 中 文 摘 要 --- p.v / Publications --- p.ix / Acknowledgements --- p.xii / List of Abbreviations --- p.xiii / Index for Figures --- p.xvi / Index for Tables --- p.xviii / Chapter Chapter 1. --- Introduction and Literature Review --- p.1 / Chapter 1.1 --- Long Bone Fracture - A Growing Global Challenge --- p.2 / Chapter 1.2 --- Long-Bone Fracture - Current Management and Limitations --- p.5 / Chapter 1.3 --- Cellular Biology of Fracture Healing --- p.7 / Chapter 1.3.1 --- Stage 1: Inflammation --- p.7 / Chapter 1.3.2 --- Stage 2: Soft callus formation --- p.8 / Chapter 1.3.3 --- Stage 3: Hard callus formation --- p.9 / Chapter 1.3.4 --- Bone Remodeling --- p.10 / Chapter 1.4 --- Biophysical Stimulation to Bone --- p.13 / Chapter 1.5 --- Low-intensity Pulsed Ultrasound --- p.14 / Chapter 1.5.1 --- Application of LIPUS on Fracture Healing --- p.14 / Chapter 1.5.2 --- Physics of Ultrasound --- p.16 / Chapter 1.5.3 --- Ultrasound Parameters --- p.20 / Chapter 1.5.3.1 --- Ultrasound Frequency --- p.20 / Chapter 1.5.3.2 --- Duty Cycle --- p.22 / Chapter 1.5.3.3 --- Intensity --- p.22 / Chapter 1.5.3.4 --- Angle of Incidence --- p.24 / Chapter 1.5.3.5 --- Ultrasound Field --- p.25 / Chapter 1.5.4 --- Possible Mechanism of LIPUS on Tissue --- p.31 / Chapter 1.5.4.1 --- Thermal Effect --- p.31 / Chapter 1.5.4.2 --- Cavitation --- p.31 / Chapter 1.5.4.2 --- Acoustic Streaming --- p.32 / Chapter 1.5.4.3 --- Frequency Resonance Hypothesis --- p.32 / Chapter 1.5.4.4 --- Micromotion --- p.33 / Chapter 1.6 --- Possible Cellular and Molecular Mechanotransduction Mechanism of LIPUS --- p.34 / Chapter 1.6.1 --- Osteocyte: Potential Mechanosensor --- p.34 / Chapter 1.6.3 --- Osteocyte-osteoblast mechanotransduction --- p.39 / Chapter 1.7 --- Hypothesis --- p.39 / Chapter 1.8 --- Study Plan and Objectives --- p.40 / Chapter 1.8.1 --- Study Plan --- p.40 / Chapter 1.8.2 --- Objectives --- p.42 / Chapter Chapter 2. --- Characterization of Ultrasound Field Distances on Rat Fracture Model --- p.43 / Chapter 2.1 --- Introduction --- p.44 / Chapter 2.2 --- Material & Methods --- p.47 / Chapter 2.2.1 --- Closed Femoral Shaft Fracture Model in Rat --- p.47 / Chapter 2.2.2 --- Ultrasound Field Distances Setup --- p.51 / Chapter 2.2.3 --- Animal Grouping & LIPUS Treatment Protocol --- p.53 / Chapter 2.2.4 --- Assessments --- p.56 / Chapter 2.2.4.1 --- Radiological Analysis --- p.56 / Chapter 2.2.4.2 --- Micro-computed Tomography --- p.61 / Chapter 2.2.4.3 --- Histomorphometry --- p.64 / Chapter 2.2.4.4 --- Mechanical Testing --- p.66 / Chapter 2.2.4.5 --- Statistical Analysis --- p.66 / Chapter 2.3 --- Results --- p.68 / Chapter 2.3.1 --- Radiological Analysis --- p.71 / Chapter 2.3.2 --- MicroCT --- p.77 / Chapter 2.3.3 --- Histomorphometry --- p.82 / Chapter 2.3.4 --- Mechanical Testing --- p.85 / Chapter 2.4 --- Discussion --- p.87 / Chapter 2.4.1 --- Far Field LIPUS Enhances Mechanical Properties of Healing Callus --- p.88 / Chapter 2.4.2 --- Mid-near field and Near field LIPUS Enhances Woven Bone Formation --- p.90 / Chapter 2.4.3 --- The Biological Effects of LIPUS with Different Ultrasound Field Exposure --- p.94 / Chapter 2.5 --- Conclusion --- p.97 / Chapter Chapter 3. --- The Effect of Ultrasound Field Distances on Bone Cells --- p.100 / Chapter 3.1 --- Introduction --- p.101 / Chapter 3.2 --- Material & Methods --- p.102 / Chapter 3.2.1 --- Cell Culture --- p.102 / Chapter 3.2.2 --- Ultrasound Field Distances Setup & Treatment Protocol --- p.102 / Chapter 3.2.2 --- Immunostaining of β-catenin --- p.106 / Chapter 3.2.3 --- Wound Healing Assay --- p.109 / Chapter 3.2.4 --- BrdU Cell proliferation Assay --- p.111 / Chapter 3.2.5 --- Alkaline phosphatase activity assay --- p.112 / Chapter 3.2.6 --- Alizarin calcium nodule staining --- p.113 / Chapter 3.2.7 --- CM characterization - PGE₂ ELISA assay --- p.114 / Chapter 3.2.8 --- CM characterization - nitrite assay --- p.114 / Chapter 3.2.9 --- Statistical Analysis --- p.115 / Chapter 3.3 --- Results --- p.116 / Chapter 3.3.1 --- Immunostaining of β-catenin --- p.116 / Chapter 3.3.2 --- Wound healing assay --- p.119 / Chapter 3.3.3 --- BrdU Cell proliferation Assay --- p.119 / Chapter 3.3.4. --- Alkaline phosphatase activity assay --- p.122 / Chapter 3.3.5 --- Alizarin calcium nodule staining --- p.122 / Chapter 3.3.6. --- CM characterization - PGE₂ ELISA assay --- p.125 / Chapter 3.3.7 --- CM characterization - nitrite assay --- p.125 / Chapter 3.4 --- Discussion --- p.128 / Chapter 3.4.1 --- The Osteogenic Effect of Far Field LIPUS-Osteocyte Conditioned Medium --- p.128 / Chapter 3.4.2. --- Mechanisms of Mechanotransduction between Osteocyte and Osteoblast --- p.131 / Chapter 3.5 --- Conclusion --- p.136 / Chapter Chapter 4. --- Rat Fracture Exposed to Far Field LIPUS by Modulating Ultrasound Transducer Diameter --- p.139 / Chapter 4.1 --- Introduction --- p.140 / Chapter 4.2 --- Material & Methods --- p.143 / Chapter 4.2.1 --- Closed Femoral Shaft Fracture Model in Rat --- p.143 / Chapter 4.2.2 --- Ultrasound Field Distances Setup & Treatment Protocol --- p.143 / Chapter 4.2.3 --- Assessments --- p.148 / Chapter 4.2.3.1 --- Radiological Analysis --- p.148 / Chapter 4.2.3.2 --- Micro-computed Tomography --- p.148 / Chapter 4.2.3.3 --- Histomorphometry --- p.149 / Chapter 4.2.3.4 --- Mechanical Testing --- p.151 / Chapter 4.2.3.5 --- ex vivo Temperature Measurements --- p.151 / Chapter 4.2.3.6 --- Statistical Analysis --- p.151 / Chapter 4.3 --- Results --- p.152 / Chapter 4.3.1 --- Radiological Analysis --- p.154 / Chapter 4.3.2 --- MicroCT --- p.157 / Chapter 4.3.3 --- Histomorphometry --- p.160 / Chapter 4.3.4 --- Mechanical Testing --- p.166 / Chapter 4.3.5 --- ex vivo Temperature Measurement --- p.168 / Chapter 4.4 --- Discussion --- p.170 / Chapter 4.4.1 --- Far field LIPUS Setup by Transducer Diameter Modulation Enhanced Fracture Healing --- p.170 / Chapter 4.4.2 --- Fractures Exposed to Far Field LIPUS with Higher Intensity Did Not Heal Better --- p.174 / Chapter 4.4.3 --- Biphasic Effect of LIPUS Intensities on Fracture Healing --- p.176 / Chapter 4.5 --- Conclusion --- p.178 / Chapter Chapter 5. --- Conclusion --- p.179 / Chapter 5.1 --- Differential Biological Effects of Ultrasound Fields --- p.180 / Chapter 5.2 --- Far Field LIPUS exposure can be achieved by transducer diameter modulation --- p.181 / Chapter 5.3 --- Biphasic Effect of Ultrasound Intensities on Fracture Healing --- p.181 / Chapter 5.4 --- Mechanotransduction between Osteocyte and Osteoblastic cells --- p.182 / Chapter 5.5 --- Clinical Implications --- p.183 / Chapter 5.6 --- Future Investigations --- p.184 / Chapter 5.7 --- Limitations --- p.184 / Bibliography --- p.186 / Appendix --- p.208

Every year over 20,000 [3] people die as a result of being in a fire. Although flames have the biggest visual impact, it is usually the smoke produced by the combustion of natural and synthetic materials that causes more damage and claims more lives. The main constituents of smoke, both the particulate matter as well as the hot and toxic gasses, are devastating to the tracheal and lung tissues. The damage caused to the lung and trachea by inhaling this smoke can increase a fire victim's susceptibility to infectious disease significantly [1]. Between 20% and 50% of people who suffer inhalation injury contract pneumonia due to the weakened status of their body's defenses [2] and between 4,800 and 6,400 [1] people die from either pneumonia or other complications. Despite the importance of the inner-lining of the trachea to a burn victim's health and survival, current treatments consist of keeping the patient in a clean environment, supplying fresh oxygen, keeping the airways open, and letting the patient's body heal itself [1]. This treatment is not so much an active healing mechanism; rather it is a passive means of allowing the body to repair itself. The main goal of this work is to develop a minimally invasive technique that will replace lost cells on the inside surface of the trachea as efficiently as possible, actively healing the patient's injury. Ideally, the patient would receive a single treatment and then make a complete recovery on his or her own. The main challenge lies in delivering an even layer of intact cells to the inner-surface of the trachea in such a manner that they will stay in place and will replace the damaged or missing tissue. The overall approach is to spray a suspension, composed of epithelial cells in an aqueous solution of Pluronic F-127 polymer, onto the trachea using a jet atomizer. Because Pluronic F-127 solutions can be liquids at room temperature but gels at body temperature, the role of the polymer will be to immobilize the cells onto the tracheal surface long enough for them to attach and grow.

Submitted by Eunice Novais (enovais@uepg.br) on 2018-03-05T18:23:35Z No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Leandro Tullio.pdf: 1410480 bytes, checksum: b92ca948d0810d08208334b6703b86c9 (MD5) / Made available in DSpace on 2018-03-05T18:23:35Z (GMT). No. of bitstreams: 2 license_rdf: 811 bytes, checksum: e39d27027a6cc9cb039ad269a5db8e34 (MD5) Leandro Tullio.pdf: 1410480 bytes, checksum: b92ca948d0810d08208334b6703b86c9 (MD5) Previous issue date: 2016-02-26 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / O pH é o principal determinante da diversidade microbiana do solo. Rhizobium freirei é um notável endossimbionte de feijão e reconhecido por sua elevada capacidade de tolerância a estresses ambientais, como o pH ácido. O objetivo deste trabalho foi analisar alterações na expressão gênica, em nível transcricional e traducional, de R. freirei cultivada em pH ácido. A bactéria foi cultivada em caldo TY até alcançar a fase exponencial em pH 6,8 e pH 4,8 e, submetida à extração de proteínas totais. As proteínas foram separadas por eletroforese bidimensional e os perfis proteicos obtidos foram analisados em imagens de alta resolução. Os spots diferencialmente expressos (p ≤ 0,05) foram selecionados para análise em espectrometria de massa e identificação in silico. A expressão diferencial dos genes gshB, gstA e ropB foi analisada após 0, 30 e 60 min de exposição de R. freirei ao pH ácido. De acordo com os resultados, o principal mecanismo de tolerância de R. freirei ao pH ácido parece estar associado à alteração de vias metabólicas centrais, devido ao aumento na expressão de proteínas envolvidas com vias oxidativas (ZwF, KDPG e PckA), relacionadas ao Ciclo do Ácido Tricarboxílico. A maior expressão destas enzimas sugere um aumento no consumo de carboidratos e consequente formação de NADH, que pode ser corroborado pelo aumento na expressão da subunidade NuoC do Complexo 1 da cadeia respiratória, que cataliza a conversão de NADH em NAD+ acoplada ao transporte de prótons para o periplasma, reduzindo a acidificação citosólica. O Complexo 1 é considerado o principal formador de radical superóxido, além disso a maior expressão das enzimas antioxidativas AhpC e GstA em nível traducional, bem como a maior expressão de gstA em nível transcricional, sugere fortemente que o pH ácido induz ao estresse oxidativo em R. freirei. Por outro lado, a menor expressão da proteína AccC pode estar relacionada a um direcionamento do metabolismo central ao consumo de ácidos em detrimento à biossíntese de ácidos graxos, processo em que AccC é fundamental. A menor expressão da subunidade β do Complexo ATPase (AtpD) pode estar direcionada à redução na acidificação do pH interno, por meio da diminuição na entrada de prótons. Tal abrangência de vias metabólicas envolvidas sugere que a resposta adaptativa ao pH ácido possui caráter multigênico, cuja predominância de proteínas citoplasmáticas reforça a importância do metabolismo central para sua capacidade de tolerar pH acidificado. Ao contrário de trabalhos que enfocam genes ou mecanismos específicos de tolerância, nossos resultados mostraram, pela primeira vez, que o metabolismo central parece ser o principal mecanismo de tolerância ao pH ácido em R. freirei e que há fortes indícios da correlação entre estresse ácido e oxidativo. / The pH is the main controller of microbial diversity of soil. Rhizobium freirei is a remarkable bean endosymbiont and recognized by its ability to tolerate environmental stresses, such as acidic pH. The aim of this study was to analyze changes in gene expression at transcriptional and translational level of R. freirei grown in acidic pH. Bacteria was grown in TY medium until exponential phase in two treatments: pH 6,8 and pH 4,8. The protein extraction was performed and the extract was separated by two-dimensional electrophoresis. High-resolution images of the profiles were analyzed and the spots that showed statistical difference in the relative volume (%vol) between treatments were selected for identification by mass spectrometry (MS) and in silico identification. Differential expression of R. freirei was assessed by analysis of gshB, gstA and ropB genes after 0, 30 and 60 min of exposure in acidic pH. According to the results, the changing of central metabolic pathways seems to be the main mechanism of tolerance to acid stress in R. freirei, due to the increased expression of proteins involved in oxidative pathways (Zwf, aldolase KDPG and PckA) related to the Tricarboxilic Acid cicle (TCA). The increased expression of these proteins suggests higher carbohydrate consumption and NADH formation, reinforced by elevated expression of NuoC subunit of respiratory chain Complex 1, that converts NADH into NAD+ coupled to proton transport to periplasm and consequent decreases the cytosolic acidification. The Complex 1 of the respiratory chain is the main superoxide former; moreover the higher expression of the antioxidative enzymes AhpC and GstA at translational level, and the higher expression of gstA at transcriptional level, strongly suggests an oxidative stress induced by acid pH condition. On the other hand, the decreased AccC protein expression supports the trend of the central metabolism to acid-consuming reactions, in contrast to fatty acids biosynthesis pathways, where AccC is essential. Down-regulation of the β subunit of ATPase Complex can be related to regulation of cytosolic acidity by reduction of proton entry through ATPase Complex. Such broad range of pathways involved in the response to acid conditions suggests an adaptive response of multigenic character mainly involving cytoplasmic proteins, and reinforces the importance of the central metabolism for R. freirei ability to tolerate acidic pH conditions. In contrast to studies focused in few genes or specific mechanisms of tolerance, our results shows by the first time that the central metabolism appears to be the main mechanism of acid tolerance in R. freirei and suggests a correlation between acid and oxidative stress responses.