Made available in DSpace on 2014-06-11T19:22:16Z (GMT). No. of bitstreams: 0 Previous issue date: 2012-02-24Bitstream added on 2014-06-13T19:48:26Z : No. of bitstreams: 1 figueiredo_pg_me_botfca.pdf: 864751 bytes, checksum: 77b495899b271a3df8c49ee115f500a4 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Características físicas do solo como porosidade, retenção de água, densidade e resistência a penetração são influenciadas diretamente pelo tipo de preparo do solo. Em resposta a estas modificações ocorrem adaptações morfofisiológicas das raízes. Verifica-se na literatura relatos sobre a influência de diferentes tipos de preparos do solo na produção e na qualidade culinária de raízes de mandioca cultivadas em diferentes tipos de solo, entretanto são desconhecidos os fatores, pertinentes às raízes, que determinam essas diferenças, o que leva a hipótese de que estes fatores possam ser de ordem morfo-anatômica. Diante disso, objetivou-se através deste trabalho de pesquisa estudar a influência de três tipos de preparo do solo (cultivo mínimo, plantio convencional e plantio direto) nas características físicas do solo, produção, morfologia e anatomia de raízes tuberosas de Manihot esculenta Crantz cultivar IAC 576-70. Para isso foi conduzido um experimento a campo em delineamento experimental de blocos ao acaso com parcelas subdivididas e seis repetições. Foram consideradas as causas de variação os tratamentos, preparos do solo, a saber: cultivo mínimo (CM), preparo convencional (PC) e plantio direto (PD) e a idade da planta, em dias após o plantio (DAP). No delineamento experimental os preparos do solo foram considerados parcelas e as idades da planta ou épocas de amostragens as subparcelas, neste experimento foram avaliados a micro, macro e porosidade total, teor de água e resistência a penetração do solo, número, comprimento, diâmetro, massas fresca e seca, porcentagem... / Soil physical characteristics such as porosity, soil water content, bulk density and penetration resistance are directly affected by tillage. In response to these changes occur roots’ morpho-physiological adaptations. In the literature there are reports about the influence of different tillage on the cassava root yields and culinary qualities affected by soil types. However the factors inner roots which determine these differences are unknown, which leads to the hypothesis that these factors may be morphological or anatomical. So the objective of this work was to study the influence of three tillage types (minimum tillage, conventional tillage and no tillage) on soil physical properties, yield, morphology and anatomy of Manihot esculenta Crantz, cultivar IAC 576-70 roots. For those was conducted a field experiment, which experimental design was randomized block in split-plot with six replications. That were considered variation causes, the treatments: minimum tillage (MT), conventional tillage (CT) and no tillage (NT) and plant age, in days after planting (DAP). In the experiment the treatments, tillages, were considered the plots and the subplots were established by plant ages or times of sampling (DAP). In this experiment were evaluated micro, macro and total porosity, soil water content and soil penetration resistance, number, length, diameter, fresh and dry weight, percentage of dry mass, morphological and anatomical characteristics and the proportion of tissues that make up the cassava tuberous roots at different ages of the plant. From the results of these evaluations there was possible to conclude that he order of setting of yield components of sweet cassava cv. IAC 576-70 are independent of tillage, and are defined in the following order: number of root, length, diameter, fresh weight, dry weight... (Complete abstract click electronic access below)

Thesis (M.S.)--Massachusetts Institute of Technology, Dept. of Electrical Engineering and Computer Science, 1981. / MICROFICHE COPY AVAILABLE IN ARCHIVES AND ENGINEERING. / Includes bibliographical references. / by Andrew Emil Yagle. / M.S.

Orientador: Fredererico Canato Martinho / Co-orientador: Márcia Carneiro Carneiro Valera / Banca: Ana Paula Martins Gomes / Banca: Izabel Coelho Gomes Camões / Resumo: Os objetivos do presente estudo são: 1) Estudar o perfil inflamatório das lesões periapicais de dentes com infecções endodônticas primárias através dos níveis das citocinas inflamatórias: IL-1beta, IL-6, TNF-alfa; correlacionando com a presença de sintomatologia clínica; 2) Investigar os níveis das metaloproteinases [MMP-1, -2 e -9], inibidores de metaloproteinase [TIMP-1 e -2] e complexos MMP/TIMP [MMP1/TIMP1, MMP1/TIMP2, MMP2/TIMP1, MMP2/TIMP2, MMP9/TIMP1 e MMP9/TIMP2], presentes nos tecidos periapicais de dentes com infecções endodônticas primárias e lesão periapical; correlacionando com a presença de sintomatologia clínica e reabsorção óssea; 3) Correlacionar os níveis das citocinas inflamatórias [IL-1beta, IL-6 e TNF-alfa] e seus antagonistas [IL1-RA, IL6-RA, sTNF-R1], presentes nos tecidos periapicais; 4) Estudar a efetividade de diferentes medicações intracanal no controle do processo inflamatório periapical, através do monitoramento dos níveis de citocinas inflamatórias, [IL-1beta, IL-6, TNF-alfa], MMP's [MMP-1, -2 e -9], TIMP's [TIMP-1 e -2] e complexos [MMP1/TIMP1, MMP1/TIMP2, MMP2/TIMP1, MMP2/TIMP2 MMP9/TIMP1, MMP9/TIMP2] coletadas dos tecidos periapicais. Vinte dentes unirradiculares com necrose pulpar e lesão periapical foram selecionados. Após o preparo biomecânico (PBM) os dentes foram divididos aleatoriamente em 2 grupos de acordo com a medicação intracanal utilizada: Ca(OH)2+SSL [Ca(OH)2 + solução salina fisiológica]; e Ca(OH)2+CLX [Ca(OH)2 + clorexidina gel 2%]. Para a análise dos biomarcadores inflamatórios, foram realizadas coletas utilizando cones de papel estéril/apirogênico, ultrapassando 2 mm além ápice para obtenção do fluído intersticial. As coletas foram realizadas em dois momentos operatórios : coleta após PBM (Coleta inicial-CIF1) e coleta após 14 dias de medicação intracanal (Coleta final-CFF2). Citocinas... / Abstract: The aim of this study was: 1) To investigate the inflammatory profile of apical periodontitis disease present in primarily infected root canals determined by the levels of inflammatory cytokines : IL-1beta, IL-6, TNF-alfa; and to correlate with the presence of clinical features; 2) To investigate the levels of metalloproteinases [MMP-1, -2, -9], metalloproteinase inhibitors [TIMP-1 e -2] ; complexes MMP/TIMP [MMP1/ TIMP1, MMP1/TIMP2, MMP2/TIMP1, MMP2/TIMP2, MMP9/TIMP1 e MMP9/TIMP2], present in apical peridontitis disease from teeth with primary endodontic infection; and to correlate with the presence of clinical features; 3) To correlate the levels of IL-1beta, IL-6, TNF-alfa with their respectively antagonists - IL1-RA, IL6-RA, s TNF-R1, all present in apical peridontitis; 4) To evaluate the effectiveness of different root canal medications in the inflammatory process by monitoring the levels of inflammatory cytokines [IL-1beta, IL-6, TNF-alfa], MMP's [MMP-1,-2,-9]; TIMP [TIMP-1 and -2] and complexes [MMP1/TIMP1, MMP1/TIMP2, MMP2/TIMP1, MMP2/TIMP2, MMP9/TIMP1 e MMP9/TIMP2] in apical periodontitis. Twenty patients with single-rooted teeth that had necrotic pulp and periapical periodontitis will be selected. After BMP teeth will be randonly devided into 2 groups according to the root canal medication selection: Ca(OH)2+SSL [Ca(OH)2 + Saline Solution]; e Ca(OH)2+CHX [Ca(OH)2 + chlorhexidine gel 2%]. Samples will be collected from intersticial fluid by using sterile/ apyrogenic paper points. Samples will performed at two diferente times: After chemomechaical preparation (S1) and after 14 of root canal medication (S2). Inflammatory cytokines, metalloproteinases and their antagonists will be dosage by ELISA-assay. The levels of inflammatory cytokines, metalloproteinases and their antagonists will be typed on a spreadsheet and statiscally analyzed by SPSS for Windows and Statistic packpage 9.0. ... / Mestre

Delia radicum L. (Diptera: Anthomyiidae), the cabbage root fly, is a specialist root-feeding insect pest of Brassica crops. The impending withdrawal of chlorpyrifos, one of the main pesticides used against D. radicum, opens new opportunities to research alternative pest management strategies. Manipulating host plant location cues to influence D. radicum adult and larval behaviour, along with induced plant defence responses, offer potential integrative crop protection solutions (Chapter 1). This thesis aimed to identify the semiochemistry underpinning D. radicum larval host plant location, and to investigate whether plant defence induction treatments (methyl jasmonate [MeJA], D-Fructose) and a herbivore induced volatile (dimethyl disulfide [DMDS]) affect D. radicum larval performance and adult oviposition preference. In choice-test bioassays, larvae were inconsistent in their responses to root exudates collected from Brassica host plants (Chapter 2). A combined bioassay and EthoVision® video-tracking approach was developed to record and analyse larval movements in response to volatiles emitted from host and non-host plant roots (Chapter 3). Larvae were significantly attracted to host plant root volatiles. Olfactory stimuli from roots of the non-host plant onion (Allium cepa L. ‘Ailsa Craig’), which share overlapping, yet distinctive volatile profiles to that of Brassica plants, also elicited positive taxis. By analysing the volatile metabolome of broccoli (B. oleracea L. convar. botrytis L. Alef. var. cymosa Duchesne ‘Parthenon’) and onion roots using solid phase micro extraction-gas chromatography-mass spectrometry (SPME-GCMS), a suite of candidate volatile orientation cues were identified. A SPME-based method was developed to non-invasively collect root volatiles in situ from glasshouse- and field-grown broccoli plants pre- and post-D. radicum infestation (Chapters 4, 5 and 6). GC-MS analyses revealed that sulfur compounds, showing characteristic temporal emission patterns, were the principal volatiles released by roots in response to damage. This new method, which has potential for wide application in chemical ecology research, allows the study of volatiles in the soil in situ that are critical for interactions between trophic levels. In EthoVision® bioassays, a major volatile constituent of broccoli roots, DMDS, was attractive to larvae, but toxic at the highest dose tested (Chapter 3). Glasshouse and on-farm experiments using broccoli were conducted to evaluate the efficacy of MeJA, D-Fructose and DMDS against D. radicum compared to commercially available crop protection products (chlorpyrifos [Dursban® WG], spinosad [Tracer®], Steinernema feltiae Filipjev [Nematoda: Steinernematidae] [Entonem] and garlic granules [ECOguard®]). MeJA and garlic reduced larval performance under glasshouse conditions whereas D-Fructose and DMDS did not at the concentrations tested (Chapter 7). In field studies, MeJA combined with reduced rate chlorpyrifos, spinosad, and S. feltiae all showed partial efficacy for controlling D. radicum larvae. Inherent field site, weather and D. radicum population density variability highlighted that glasshouse results cannot always be reproduced in more complex field environments (Chapter 8). Further research is needed into formulation, mode of application and timing to improve efficacy of promising treatments that may help in future integrated pest management (IPM) for this key pest in the absence of existing pesticides.

A presença de projeção dentinária acima da linha de término no preparo de dentes tratados endodônticamente para receber núcleos metálicos fundidos é bem fundamentada na literatura, influenciando positivamente na resistência da raiz. Entretanto, devido à destruição dental, nem sempre é possível atingir uma forma de projeção homogênea, contribuindo para deficiências no preparo que podem influenciar na resistência mecânica à fratura. Um incisivo central superior direito foi scaneado com tomografia computadorizada e reconstruído com software específico em um modelo virtual. Este modelo foi editado com diferentes projeções dentinárias, 2 mm acima da linha de término envolvendo projeções: ausente, total, vestibular, palatina e proximal e submetidas à simulação de uma carga mastigatória, no software Ansys Workbench v11, para avaliar a distribuição de tensões geradas no mesmo. As simulações demonstram que os melhores resultados foram obtidas com a projeção total, a projeção palatina obteve resultados muito próximos à total. O pior resultado foi da projeção ausente obtendo resultados muito próximos á projeção vestibular. Os autores nesta pesquisa concluíram que uma projeção dentinária entre o núcleo e a força aplicada é um importante fator para beneficiar a biomecânica do remanescente dental, e que a projeção total fornece o melhor prognóstico nesta situação comparando às diferentes condições. / The presence of dentin projection above the finish line in the preparation of endodonticaly treated tooth to receive post and core is well established in literature, it influences positively in the resistance of the root. However, due to dental destruction, it\'s not always possible to achieve a homogenous projection design, contributing for deficiencies in the preparation that can influence in the mechanical resistance to breaking. A maxillary right central incisor was scanned with computer tomography and reconstructed with specific software into a virtual model. This model was edited with different dentin projections, 2 mm above the preparation line involving absent, total, bucal, palatal and proximal projections and later submitted to simulation of chewing load, in software ANSYS v11, to get the distribution of tensions generated in the same. The simulations show that the best results are when the dentin projection is total, followed closely by the palatal projection. The worse result belongs to the absent projection group followed closely by bucal group. The authors in this research concludes that a dentin projection between the core and the applied force is an important factor to benefit the biomechanical of the remaining anterior teeth, and the whole projection gives the best prognostic comparing the different situations.

O objetivo deste estudo foi investigar in vitro a influência do tratamento de superfícies radiculares na adesão e proliferação de fibroblastos gengivais e de ligamento periodontal humano em fragmentos radiculares de dentes humanos extraídos por razões periodontais. Todos os fragmentos receberam raspagem e alisamento radicular (RAR), e em seguida aleatoriamente divididos em grupos, de acordo com a substância utilizada no tratamento de superfície (n= 15/grupo): ácido fosfórico 37% aplicado por 90s (AF90) ou 180s (AF180) (RAR); EDTA 24% aplicado por 90s (EDTA90) ou 180s (EDTA180); ácido cítrico pH 1.0 a 10% aplicado por 90s (AC90) ou 180s (AC180); ácido cítrico pH 1.0 a 10% associado à tetraciclina 50% por 90s (ACTC90) ou 180s (ACTC180); tetraciclina hidroclorídrica (50mg/ml) aplicada por 90s (TC90) ou 180s (TC180). O grupo controle foi composto por fragmentos tratados por meio de RAR seguido de lavagem em soro fisiológico. Após a realização dos tratamentos, os espécimes (n=3/grupo) foram preparados para análise em MEV com o objetivo de avaliar a descontaminação das superfícies radiculares por meio dos índices de rugosidade superficial (IRS), cálculo residual (ICR), perda de substância dentária (IPSD), presença de restos teciduais (IPRT), remoção de smear layer (IRSL), abertura dos túbulos dentinário (ATD) e smear layer remanescente (SLR) em fotomicrografias em aumentos de 500x e 1000x. Em 6 espécimes de cada grupo, foram plaqueados 104 fibroblastos gengivais (FGH-1), e em outros 6, 104 fibroblastos de ligamento periodontal humano (FLP-1). Após 24h foram fixados 6 espécimes por grupo (n=3/grupo) e após 48h os outros 6 espécimes (n=3/grupo), para análise em microscópio eletrônico de varredura. Para determinar a adesão e proliferação celular, o número de células aderidas à superfície nos dois períodos de avaliação foi determinado em triplicatas por um examinador independente A comparação entre os grupos foi realizada pelo método Kruskal-Wallis complementado pelo teste de Dunn para variáveis não lineares, e por meio de análise de variância múltipla (ANOVA) complementado pelo teste de Tukey para as variáveis lineares. A comparação entre os pares nos períodos de 24 e 48 horas foi realizada por meio do método ANOVA pósteste Sidak para variáveis lineares, e Kruskal Wallis pós-teste Dunn para variáveis não lineares. Foi adotado nível de significância de 5% em todos os testes. Não houve diferença estatisticamente significante para IRS, IPSD, IPRT, IRSL, ATD e SLR. Houve maior quantidade de cálculo residual nos grupos TC90 (3,66 ± 0,57; mediana = 4) e AF180 (3,66 ± 0,57; mediana = 4), enquanto que o grupo AC90 (1,33 ± 0,57; mediana = 1) mostrou quantidade significativamente menor de cálculo residual. Encontrou-se uma adesão significativamente maior de células FGH-1, no grupo EDTA180 (170 ± 77,99) no período de 24 horas, e maior efeito proliferativo (48 horas) no grupo TC90 (172,90 ± 65,38). Para células FLP-1, observou-se uma adesão significativamente maior no grupo ACTC90 (74,67 ± 98,84) no período de 24 horas e maior efeito proliferativo (48 horas) no grupo AC90 (173,8 ± 139,6). A partir dos resultados obtidos, sugere-se que a substância a ser utilizada para o condicionamento das superfícies radiculares seja escolhida de acordo com os objetivos do tratamento periodontal: quando se objetiva a formação de nova inserção conjuntiva o uso do EDTA ou AF por 180s ou da tetraciclina por 90s; para regeneração dos tecidos periodontais, sugere-se o uso de AC por 90s. / The aim of this study was to evaluate in vitro the influence of root surface conditioning on adhesion and proliferation of gingival and periodontal ligament fibroblasts on human root fragments of teeth extracted for periodontal reasons. Fragments received scaling and root planning (SRP), and were then randomly allocated into groups according to the substance used for root surface treatment (n= 15/grupo): phosphoric acid 37% applied for 90s (PA90) or 180s (PA180); EDTA 24% applied for 90s (EDTA90) or 180s (EDTA180); 10% citric acid pH 1.0 applied for 90s (CA90) or 180s (CA180); 10% citric acid pH 1.0 associated to tetracycline HCL 50% applied for 90s (CATC90) or 180s (CATC180); tetracycline hydrocloride (50mg/ml) applied for 90s (TC90) or 180s (TC180). Control group was composed by SRP treated root fragments, followed by saline solution washing. After treatment completion, specimens (n=3/grupo) were prepared for scanning electronmicroscopy (SEM) analysis, aiming at evaluation of its surfaces according to the following indexes: superficial roughness (SR); residual calculus (RC); loss of tooth substance (LT); tissue residual (TS), smear layer removal (SLR), dentin tubules opening (DTO) and smear layer residual (SLR) in photomicrographs on 500x and 1000x magnifications. In 6 specimens of each group 104 gingival fibroblasts (HGF-1) were plated; and over another 6 specimens, 104 periodontal ligament fibroblasts (PLF-1). After MEV evaluation, the number of cells adhered to the root surfaces over 24h and 48h were assessed by a calibrated examiner in triplicates. Groups comparison were analyzed through Kruskal-Wallis post-test Dunn for comparisons for non-linear variables, and ANOVA post-test Tuckey for linear variables. Comparison between pairs over 24 and 48 hours was accessed through Kruskal-Wallis post-test Dunn for non-linear variables, and ANOVA post-test Sidak for linear variables. Significance level of 5% was adopted in all tests. There was no statistical difference for SR, LT, TS, SLR, DTO and SLR. Although there was higher amounts of residual calculus on groups TC90 (3,66 ± 0,57; median = 4) and FA180 (3,66 ± 0,57; median = 4) while group CA90 (1,33 ± 0,57; median = 1) showed statistically less residual calculus. A singnificantlly higher HGF-1 cell count was found on EDTA180 (170 ± 77,99) on 24-hour period and a higher proliferative effect (48 hours) on group TTC90 (172,90 ± 65,38). A significantly higher cell adhesion for (PLF-1) was found on group ACTC90 (74,67 ± 98,84) at 24-hour assessment, and higher proliferative effect (48 hours) for AC90 (173,8 ± 139,6). From the data here exposed, it is suggested that the substance election for root surface conditioning should be based on the treatment primary goal: when a new connective tissue adhesion is aimed, EDTA or PA for 180s or TTC for 90s should be chosen; on the other hand, for periodontal regeneration, CA for 90s should be the best option.

A presença de projeção dentinária acima da linha de término no preparo de dentes tratados endodônticamente para receber núcleos metálicos fundidos é bem fundamentada na literatura, influenciando positivamente na resistência da raiz. Entretanto, devido à destruição dental, nem sempre é possível atingir uma forma de projeção homogênea, contribuindo para deficiências no preparo que podem influenciar na resistência mecânica à fratura. Um incisivo central superior direito foi scaneado com tomografia computadorizada e reconstruído com software específico em um modelo virtual. Este modelo foi editado com diferentes projeções dentinárias, 2 mm acima da linha de término envolvendo projeções: ausente, total, vestibular, palatina e proximal e submetidas à simulação de uma carga mastigatória, no software Ansys Workbench v11, para avaliar a distribuição de tensões geradas no mesmo. As simulações demonstram que os melhores resultados foram obtidas com a projeção total, a projeção palatina obteve resultados muito próximos à total. O pior resultado foi da projeção ausente obtendo resultados muito próximos á projeção vestibular. Os autores nesta pesquisa concluíram que uma projeção dentinária entre o núcleo e a força aplicada é um importante fator para beneficiar a biomecânica do remanescente dental, e que a projeção total fornece o melhor prognóstico nesta situação comparando às diferentes condições. / The presence of dentin projection above the finish line in the preparation of endodonticaly treated tooth to receive post and core is well established in literature, it influences positively in the resistance of the root. However, due to dental destruction, it\'s not always possible to achieve a homogenous projection design, contributing for deficiencies in the preparation that can influence in the mechanical resistance to breaking. A maxillary right central incisor was scanned with computer tomography and reconstructed with specific software into a virtual model. This model was edited with different dentin projections, 2 mm above the preparation line involving absent, total, bucal, palatal and proximal projections and later submitted to simulation of chewing load, in software ANSYS v11, to get the distribution of tensions generated in the same. The simulations show that the best results are when the dentin projection is total, followed closely by the palatal projection. The worse result belongs to the absent projection group followed closely by bucal group. The authors in this research concludes that a dentin projection between the core and the applied force is an important factor to benefit the biomechanical of the remaining anterior teeth, and the whole projection gives the best prognostic comparing the different situations.

Chlorosis in plants has been recognized as a devastating disease for over one-hundred and fifty years. It is easily recognized by a yellowing of the plant foilage and is associated with a reduced chlorophyll synthesis. Chlorosis is found so frequently on calcareous soils that its cause is attributed at least in part to this soil factor. It is therefore referred to as lime-induced chlorosis. This type of chlorosis has become a serious problem, especially in many of the fruit-growing regions of Europe and the Western United States where the soils are calcareous. In Utah the disease is more destructive than any other nutritional disease that affect horticultural crops. The exact cause of lime-induced chlorosis is not known, nor has a satisfactory control been developed as yet. Many conditions found in high-lime soils have been studied with regard to chlorosis, yet the problem is not so simple as it may appear. Green and chlorotic plants are frequently found growing in the same field, and yet a chemical analysis of the two soils fails to indicate a significant difference between them. It is common in chlorotic orchards to see trees where some of the major branches produce chlorotic foilage while the remainder of the tree is healthy and green. The severity of chlorosis varies from tree to tree as well as from season to season. It has been found that the heavier textured soils which are poorly drained are more conductive to chlorosis than are the lighter textured soils. If the soil moisture is held near field capacity, chlorosis is much more severe than if the soil is allowed to approach the wilting point before irrigation. Recent studies have also shown that the bicarbonate ion in solution cultures can induce chlorosis and retard the uptake of iron by plants. Since these factors--high moisture, soil aeration, and bicarbonate ion concentration--are all interrelated, it seems that their relationship to chlorosis should be investigated more thoroughly. Workers in the past have failed to show a consistent correlation between either the oxygen or the carbon dioxide can be measured at the soil-root interface. this is after all the only place in the soil that is of great importance as far as the living plant is concerned. Since the severity of chlorosis increases under poorly aerated conditions on calcareous soils, this hypothesis has been advanced: The respired carbon dioxide is given off at the plant root as carbonic acid which in a calcareous soil reacts with calcium and magnesium carbonate to give an increased bicarbonate ion concentration in the root environment. The resulting increased concentration of bicarbonate may reduce the effective oxygen at the root and thereby disturb the normal respiration and active-ion absorption of the roots or the increased concentration of bicarbonate may affect the roots absorption and plant metabolism in some other manner. The purpose of this investigation was to study the effect of the bicarbonate ion and different aeration mixtures on chlorosis. A preliminary study was made to relate the composition of the soil solution taken from the field with different conditions associated with chlorosis.

No description available.

Bacteroids of B. japonicum from nodules of soybean roots were isolated using differential centrifugation (the standard bench method) and density gradient centrifugation methods (either sucrose- or Percoll-) under anaerobic conditions in which N2 fixation was preserved. The relationships between N2 fixation and respiration, O2 supply, O2 demand, substrate (mainly malate) transport and metabolism in bacteroids were investigated using the flow chamber system. In related experiments, the primary products of N2 fixation which leave the bacteroids were investigated using a 15N-labelling technique in a closed shaken system and other biochemical methods.¶ In the flow chamber experiments, the rates at which O2 was supplied to bacteroids in the chamber were varied by (a) changing the flow rate of reaction medium through the chamber; (b) by changing the [O2 free] in the inflowing reaction medium by using either 3-5% (v/v) or 100% air in the gas mixture above the stirred reaction medium in two reservoir flasks; (c) by successively withdrawing bacteroids from the chamber, thus increasing the supply of O2 per bacteroid to those remaining in the chamber. The results showed that the rate of O2 supply regulates respiratory demand for O2 by bacteroids rather than the O2 concentration present in the reaction system. Respiration is always coupled to N2 fixation. ¶ Uptake of malate by bacteroids withdrawn from the flow chamber was measured under microaerobic conditions. Malate uptake by these N2-fixing bacteroids was lower than that by bacteroids isolated under aerobic conditions, which eliminate N2 fixation of bacteroids, but is closely correlated with bacteroid respiration rates. When respiration was increased by an increase in O2 supply, malate uptake by bacteroids was also increased. This suggested that transport of malate through the bacteroid membrane is also regulated by O2 supply, but indirectly. Higher uptake by bacteroids under aerobic conditions was observed because respiration was enhanced by the high availability of O2, but the fast uptake of malate by bacteroids driven by the abnormal respiration rates may not reflect the reality of malate demand in vivo by bacteroids when N2 fixation by bacteroids is fully coupled. ¶ The results of 15N labelling experiments and other biochemical assays once again demonstrated that ammonia is the principal significant 15N labelled product of N2 fixation accumulated during 30 min in shaken assays with 0.008-0.01 atm O2. Alanine although sometimes found in low concentrations in the flow chamber reactions, was not labelled with 15N in shaken closed system experiments. No evidence could be obtained from the other biochemical assays, either. Therefore, it is concluded that these and earlier results were not due to contamination with host cytosolic enzymes as suggested by Waters et al. (Proc. Natl. Aca. Sci. 95, 1998, pp 12038-12042). ¶ Malate transported into bacteroids is oxidized in a modified TCA cycle present in bacteroids. The results of flow chamber experiments with a sucA mutant (lacking a-ketoglutarate dehydrogenase) showed that respiratory demand for O2 by the mutant bacteroids is regulated by O2 supply in the same way as the wild-type. Despite differences in other symbiotic properties, rates of nitrogen fixation by the mutant bacteroids, based on the bacteroid dry weight, appeared to be the same as in the wild-type. Also N2 fixation was closely coupled with respiration in the same manner in both mutant bacteroids and wild type bacteroids. These results and other supporting data, strongly support the conclusion that there is an alternative pathway of the TCA cycle in bacteroids, which enables the missing step in the mutant to be by-passed with sufficient activity to support metabolism of transported malate.