Tuber aestivum is an ectomycorrhizal ascomycete with underground fruit bodies. It is an economically important species, but has been regarded as endangered in Sweden. My inventory has increased the number of reported localities from 3 to 31. It has long been debated whether T. aestivum and T. uncinatum are conspecific or not, so a clarification would help conservation biology and cultivation. My study included 117 fruit bodies of both taxa from 8 countries. The phylogenetic (ITS) and microscopic analyses showed that the two taxa were synonyms and that the spore reticulum height, used to separate the taxa, is not diagnostic. T. aestivum was clearly different from T. mesentericum, which I reported new to Sweden. The Gotland T. aestivum population was genetically distinct (RAPD) from other European specimens. The genetic variation suggested sexual reproduction. The habitat of 18 T. aestivum sites on Gotland were analysed and compared with data from France. No striking functional differences in soil chemistry were found, so a possible T. aestivum ecotype on Gotland would rather be an adaptation to the colder and drier climate. Selecting local T. aestivum inoculum for truffle orchards in Northern Europe could be important for successful truffle production. In 1999, 10 experimental truffle orchards with a total of 240 oak and hazel seedlings were established on Gotland, and as a result of this project 3000 commercial oak seedlings were planted in 2000-2001. In 2004, T. aestivum mycorrhiza was still present in all of the 22 orchards studied on Gotland, some in soils different from natural habitats. In addition, the project has also generated a truffle cultivation association, a truffle company, truffle dog breeding and export of T. aestivum to France.

We have investigated the use of deep UV laser induced fluorescence for the sensitive detection and spectroscopic lifetime studies of terbium doped dipicolinic acid (DPA-Tb) and used this to study the optical characteristics of DPA which is a chemical surrounding most bacterial spores. Background absorption spectra, fluorescence spectra, and Excitation Emission Matrix (EEM) spectra were made of the DPA-Tb complex, using both fixed 266 nm wavelength and tunable (220 nm -- 280 nm) UV laser excitations. Of importance, the fluorescence lifetimes of the four main fluorescence peaks (488 nm, 543 nm, 581 nm, and 618 nm) of the DPA-Tb complex have been measured for the first time to our knowledge. The lifetimes of all the fluorescing lines have been measured as a function of DPA-Tb concentration, solvent pH, and solvent composition, including that for the weakest fluorescing line of DPA-Tb at 618 nm. In addition, a new spectroscopic lifetime measurement technique, which we call "Transient Fluorescence Spectroscopy" was developed. In this technique, a weak, quasi-CW, amplitude modulated UV laser (8.5 kHz) was used to measure the lifetimes of the fluorescence lines, and yields insight into energy transfer and excitation lifetimes within the system. This technique is especially useful when a high power laser is not either available or not suitable. In the latter case, this would be when a high power pulsed deep-UV laser could produce bleaching or destruction of the biological specimen. In addition, this technique simulated the excitation and fluorescence emission of the DPA-Tb using a 4-level energy model, and solved the dynamic transient rate equations to predict the temporal behavior of the DPA-Tb emitted fluorescence. Excellent agreement between the experiments and the simulation were found. This technique has the potential to provide a more accurate value for the fluorescence lifetime values. In addition, with the use of asymmetric excitation waveforms, the dynamic transient rate equation analysis may allow for detailed studies of selected transfer mechanisms in a wide range of other spectroscopic applications including rare-earth solid-state lasing materials and biological samples.

OPSOMMING: 'n Literatuurstudie is onderneem oor musikale ontlening soos dit in verskillende tydperke in die verlede gemanifesteer het. 'n Literatuurstudie is ook onderneem oor die historiese verband tussen Buxtehude en Bach, musikaal en andersins. 'n Indringende ondersoek na ooreenkomste in melodiese openingsformasies in die vrye orrelwerke van Buxtehude en Bach is gedoen. Die resultate is gesistematiseer deur dit in kategorieë in te deel. Repertoriums uit voorafgaande tydperke is betrek, om daaruit gemeenskaplike historiese spore, wat verband hou met die ooreenkomste in tematiese materiaal in die vrye orrelwerke van Buxtehude en Bach, aan te dui. Vroeëre repertoriums wat betrek is, sluit byvoorbeeld Middeleeuse cantus plani, Joodse melodieë soos opgeteken deur Idelsohn, die Geneefse Psalter en vroeë klawerbordliteratuur in. Die resultate van die ontleding is gesistematiseer in 'n nuut-ontwerpte model wat gebaseer is op ooreenkomstige melodiese aanvangspatrone in die vrye orrelwerke van Buxtehude en Bach. Hierdie sistematiese model is nie slegs op die tematiese materiaal van Buxtehude en Bach van toepassing nie, maar ook op tematiese materiaal wat veer verder in die verlede teruggevind kan word, tot sover as die Delfiese himnes van 127 v.C. In hierdie studie is insig in musikale ontlening verkry uit die merkbare ooreenkomste tussen die melodiese aanvangspatrone in die tematiek van Bach se vrye orrelwerke en die van Buxtehude. Die insig is verdiep deur die sistematiese ordening van hierdie ooreenkomste, en die vasstelling dat die ooreenkomste op gemeenskaplike historiese spore teruggaan. In die tradisionele studie van musiektekstuur is harmoniese progressies gesistematiseer met die vier kadense aan die eindes van frases as uitgangspunt. 'n Ooreenkomstige sistematisering van melodiese patrone wat aan die begin van frases voorkom, is egter, sover vasgestel kon word, nog nie voorheen onderneem nie. 'n Nuwe bydrae tot die musiekwetenskap is dus gelewer deurdat die melodiese openingspatrone volgens 'n nuut-ontwerpte model gesistematiseer is. / SUMMARY: Musical borrowing in Western music is a vast field, the history of which, according to J. Peter Burkholder, still has to be written. The main objective of this study is to provide a historical perspective on musical borrowing to serve as a background for a comparative study of thematics in the free organ works of Buxtehude and Bach. However, to provide a more comprehensive perspective on the phenomenon of musical borrowing and traces from the past, one requires a historical consciousness which goes further than Bach's immediate past. The research includes a study of literature on musical borrowing as it has manifested itself in various eras. This literature study also includes an investigation into the historical connection between Buxtehude and Bach, musically and otherwise. Similarities in melodic opening formations in the free organ works by Buxtehude and Bach are pointed out. The results are systematicised by the classification into categories. The classification draws on repertoires from previous eras in order to indicate common historical traces relating to correspondences of thematic material in the free organ works by Buxtehude and Bach. Earlier repertoires include Medieval chants, Jewish melodies recorded by Idelsohn, the Genevan Psalter and early keyboard literature. The results of the analysis are systematised in a newly designed model which is based on corresponding melodic opening patterns in the free organ works by Buxtehude and Bach. This systematic model applies not only to thematic material of Buxtehude and Bach but also to thematic material which can be traced back far further into the past, in fact, as far back as the Delphic hymns of 127 BC. This study provides insight into musical borrowing based on the noticeable similarities between melodic opening phrases in the thematics of the free organ works by Bach and Buxtehude. This insight is enhanced by the systematic ordering of these similarities and the realisation that the similarities are based on common historical traces. In the traditional study of musical texture harmonic progressions are systematised, using the four cadences at the ends of phrases as the point of departure. However, melodic patterns which occur at the beginnings of phrases, have not, as far as I know, been similarly systematised. This systemisation of melodic opening patterns according to a newly designed model therefore constitutes a new contribution to research in musicology. / Thesis (Ph.D. (Music))--North-West University, Potchefstroom Campus, 2010.

OPSOMMING: 'n Literatuurstudie is onderneem oor musikale ontlening soos dit in verskillende tydperke in die verlede gemanifesteer het. 'n Literatuurstudie is ook onderneem oor die historiese verband tussen Buxtehude en Bach, musikaal en andersins. 'n Indringende ondersoek na ooreenkomste in melodiese openingsformasies in die vrye orrelwerke van Buxtehude en Bach is gedoen. Die resultate is gesistematiseer deur dit in kategorieë in te deel. Repertoriums uit voorafgaande tydperke is betrek, om daaruit gemeenskaplike historiese spore, wat verband hou met die ooreenkomste in tematiese materiaal in die vrye orrelwerke van Buxtehude en Bach, aan te dui. Vroeëre repertoriums wat betrek is, sluit byvoorbeeld Middeleeuse cantus plani, Joodse melodieë soos opgeteken deur Idelsohn, die Geneefse Psalter en vroeë klawerbordliteratuur in. Die resultate van die ontleding is gesistematiseer in 'n nuut-ontwerpte model wat gebaseer is op ooreenkomstige melodiese aanvangspatrone in die vrye orrelwerke van Buxtehude en Bach. Hierdie sistematiese model is nie slegs op die tematiese materiaal van Buxtehude en Bach van toepassing nie, maar ook op tematiese materiaal wat veer verder in die verlede teruggevind kan word, tot sover as die Delfiese himnes van 127 v.C. In hierdie studie is insig in musikale ontlening verkry uit die merkbare ooreenkomste tussen die melodiese aanvangspatrone in die tematiek van Bach se vrye orrelwerke en die van Buxtehude. Die insig is verdiep deur die sistematiese ordening van hierdie ooreenkomste, en die vasstelling dat die ooreenkomste op gemeenskaplike historiese spore teruggaan. In die tradisionele studie van musiektekstuur is harmoniese progressies gesistematiseer met die vier kadense aan die eindes van frases as uitgangspunt. 'n Ooreenkomstige sistematisering van melodiese patrone wat aan die begin van frases voorkom, is egter, sover vasgestel kon word, nog nie voorheen onderneem nie. 'n Nuwe bydrae tot die musiekwetenskap is dus gelewer deurdat die melodiese openingspatrone volgens 'n nuut-ontwerpte model gesistematiseer is. / SUMMARY: Musical borrowing in Western music is a vast field, the history of which, according to J. Peter Burkholder, still has to be written. The main objective of this study is to provide a historical perspective on musical borrowing to serve as a background for a comparative study of thematics in the free organ works of Buxtehude and Bach. However, to provide a more comprehensive perspective on the phenomenon of musical borrowing and traces from the past, one requires a historical consciousness which goes further than Bach's immediate past. The research includes a study of literature on musical borrowing as it has manifested itself in various eras. This literature study also includes an investigation into the historical connection between Buxtehude and Bach, musically and otherwise. Similarities in melodic opening formations in the free organ works by Buxtehude and Bach are pointed out. The results are systematicised by the classification into categories. The classification draws on repertoires from previous eras in order to indicate common historical traces relating to correspondences of thematic material in the free organ works by Buxtehude and Bach. Earlier repertoires include Medieval chants, Jewish melodies recorded by Idelsohn, the Genevan Psalter and early keyboard literature. The results of the analysis are systematised in a newly designed model which is based on corresponding melodic opening patterns in the free organ works by Buxtehude and Bach. This systematic model applies not only to thematic material of Buxtehude and Bach but also to thematic material which can be traced back far further into the past, in fact, as far back as the Delphic hymns of 127 BC. This study provides insight into musical borrowing based on the noticeable similarities between melodic opening phrases in the thematics of the free organ works by Bach and Buxtehude. This insight is enhanced by the systematic ordering of these similarities and the realisation that the similarities are based on common historical traces. In the traditional study of musical texture harmonic progressions are systematised, using the four cadences at the ends of phrases as the point of departure. However, melodic patterns which occur at the beginnings of phrases, have not, as far as I know, been similarly systematised. This systemisation of melodic opening patterns according to a newly designed model therefore constitutes a new contribution to research in musicology. / Thesis (Ph.D. (Music))--North-West University, Potchefstroom Campus, 2010.

Viability and dispersal of ascospores of Anisogramma anomala, the cause of eastern filbert blight (EFB) on European hazelnut, from diseased branches pruned from trees were measured. In each of two years, branches bearing stromata of A. anomala were cut in mid-December and compared to branches cut near budbreak in March, when trees became susceptible to infection. The experiment was replicated three times at separated locations. At each location, 125 diseased branches (source) were piled loosely in a 1 x 1 m area. From March to June, spore traps (rain sampling-type) as well as 2-year-old potted hazelnut trees were placed next to each source, 6.4 m upwind and downwind, and 20 m downwind from each source. During seven significant major rain events over the two seasons, hazelnut seedlings (3-month-old) were placed adjacent to the spore traps. Near sources significantly higher (P. < 0.01) ascospores counts were obtained for branches cut near budbreak compared to those pruned in December in the first season; no significant difference in counts of ascospores were observed in the second season between pruning treatments. For both seasons significantly higher (P < 0.05) counts of ascospores were observed at 6.4 m downwind compared to 6.4 m upwind or 20 m downwind of a source. Ascospore viability, as assessed by staining with trypan blue, was similar for both pruning times at all distances and averaged 50%. At least one infected seedling was obtained for 5 of 7 major rain events regardless of pruning time at sources and 3 of 7 major rain events 6.4 m downwind of a source. All of the 2-year-old potted trees for both pruning treatments at the source and 6.4 m downwind became diseased and > 50% of trees at 20 m downwind became diseased in the 2010 season. Similar to ascospores counts, disease incidence was significantly higher (P < 0.01) in 2-year-old potted trees observed 6.4 m downwind compared to 6.4 m upwind or 20 m downwind in the 2010 season. Significantly higher (P < 0.01) disease incidence in 2-year-old potted trees was observed 20 m downwind compared to 6.4 m upwind in the 2010 season. Downwind disease gradients for both pruning treatments were shallow with slopes that were not significantly different than zero (p > 0.05) for the 2010 season. Based on these results, ascospores from diseased branches pruned from trees in both pruning treatments remained viable, infectious and were dispersed downwind of each treatment. / Graduation date: 2012

The data from public herbal collections were used to determine the frequency of the occurence of fertile bracken populations. Samples of bracken were collected to assess the degree of fertility and also to reveal the frequency of both diploid and triploid cytotypes using flow cytometry. Collected samples were also used for SSR analysis to characterize the population structure of both cytotypes and the propagation of triploid plants of bracken.

SOUZA, Pedro Filho Noronha de.Purificação, caracterização bioquímica e atividade antimicrobiana de uma nova albumina 2s da torta da mamoneira (Ricinus communis L.), com atividade inibitória contra tripsina. 2012. 114 f. Dissertação (Mestrado em Bioquímica) - Universidade Federal do Ceará, Fortaleza-CE, 2012. / Submitted by Eric Santiago (erichhcl@gmail.com) on 2016-06-27T14:14:02Z No. of bitstreams: 1 2012_dis_pfnsouza.pdf: 1847174 bytes, checksum: 6675a05ab248cfb741080071e366161c (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-08-02T20:11:34Z (GMT) No. of bitstreams: 1 2012_dis_pfnsouza.pdf: 1847174 bytes, checksum: 6675a05ab248cfb741080071e366161c (MD5) / Made available in DSpace on 2016-08-02T20:11:34Z (GMT). No. of bitstreams: 1 2012_dis_pfnsouza.pdf: 1847174 bytes, checksum: 6675a05ab248cfb741080071e366161c (MD5) Previous issue date: 2012 / Castor bean (Ricinus communis L.) is an important crop for the Northeast of Brazil, which recently has been used to produce biodiesel. Around 90% of the castor bean production in Brazil is concentrated at Northeast region and the state of Ceará is the second largest producer. During the oil extraction process from the castor bean seeds, a resulting residue, called castor cake, is underutilized. However, this byproduct is rich in protein and micronutrients such as nitrogen, phosphorus and potassium, an attribute that qualifies it as a stuff that could be used as organic fertilizer or as animal feed, for example, bringing added value to it. Unfortunately, its use as food has not been possible because of the presence of toxic elements and allergens (ricin, ricinine, complex allergens) in its composition, unless it were previously submitted to a detoxification process. To date, the existing technologies to this end are not economically viable on an industrial scale. Therefore, studies to add value to this abundant byproduct generated in the castor bean biodiesel productive chain is of paramount importance. Thus, in this context, this study was performed to identify, isolate, purify and characterize new bioactive molecules of de-oiled castor cake with biotechnological potential. Through extraction of soluble proteins with 50 mM Tris-HCl pH 7.5, fractionation with ammonium sulfate (50-75%), following by hydrophobic interaction chromatography in Phenyl-Sepharose column and ion exchange chromatography (DEAE-Sepharose), a protein, named Rc-2S-Alb, able to inhibit trypsin was purified. Rc-2S-Alb has a molecular mass of approximately 75.8 kDa, as determined by SDS-PAGE, and under reducing conditions showed a large and a small protein band of 15.8 kDa and 10.5 kDa, respectively. Its NH2-terminal sequence showed similarity with the following proteins: putative 2S albumin precursor (89%), chain A of RicC3 (89%), and chain A of mabilin-1 (89%), all from R. communis seeds; and with the short chain of a "napin-like" protein from Brassica napus (89%). In addition, these similar proteins have two highly conserved domains: QEVQRKDLS and YLRQS. Comparison of the partial primary structure of Rc-2S-Alb generated by the ESI-Q-TOF MS/MS analysis of 17 tryptic peptides, showed 43% similarity to Mabinlin-1 (pI/Mr 6.7 and 29.3 kDa) from R. communis. There were also high similarities among the three-dimensional structures of Rc-2S-Alb, RicC3 and Mabinlin-1. Rc-2S-Alb did not inhibit the spore germination of the phytopathogenic fungi Fusarium oxysporum and Rizoctonia solani, but promoted aggregation of their respective spores. Moreover, Rc-2S-Alb did not inhibit the mycelial growth of Fusarium oxysporum, Fusarium solani, Rizoctonia solani and Collethotricum gloeosporioides. Contrary, Rc-2S-Alb was effective in inhibiting the growth of the human pathogenic bacteria Pseudomonas aeruginosae, Klebsiella pneumoniae and Bacillus subtilis, at low concentrations. In conclusion, it was established a protocol to purify a new 2S albumin from castor bean cake, which inhibits trypsin and has important antibacterial activity. Thus, the Rc-2S-Alb should be further studied in order to verify its effectiveness as new alternative therapeutic agent against resistant bacteria to commercial antibiotics, which will contribute to improve the human health. / A mamoneira (Ricinus communis L.) é uma cultura importante para a região Nordeste do Brasil, onde, recentemente, tem sido utilizada para produção de biodiesel. O Nordeste detém 90% da produção brasileira de mamona, sendo o Ceará, o segundo maior produtor. No processo de extração de óleo das sementes de mamoneira para produção de biodiesel, o resíduo resultante, denominado de torta da mamona, representa um subproduto pouco utilizado. Apesar disso, essa torta é rica em proteínas e micronutrientes, como nitrogênio, fósforo e potássio, atributo que a qualifica como um insumo que poderia ser utilizado como adubo orgânico, ou mesmo como ração animal, o que lhe agregaria valor comercial. Todavia, seu uso como alimento não tem sido possível por causa da presença de elementos tóxicos e alergênicos (Ricina, Ricinina, Complexos Alergênicos) na sua composição, a não ser que passasse por processamento para sua destoxificação. Infelizmente, tecnologia economicamente viável para esse fim, em escala industrial, ainda é inexistente. Portanto, há necessidade de pesquisas que venham a agregar valor a esse subproduto abundante da cadeia produtiva do biodiesel. Assim, nesse contexto, o presente trabalho está inserido num projeto cujo objetivo maior é identificar, isolar, purificar e caracterizar novas moléculas bioativas da torta delipidada de sementes de mamoeira com potencial biotecnológico. Através de extração de proteínas solúveis com tampão Tris-HCl, 50 mM, pH 7,5, e fracionamento do extrato obtido com sulfato de amônio (50-75%), cromatografia de interação hidrofóbica em coluna Phenyl-Sepharose e cromatografia de troca iônica (DEAE-Sepharose) foi possível purificar uma albumina 2S, denominada Rc-2S-Alb, capaz de inibir tripsina. A Rc-2S-Alb apresentou massa molecular de, aproximadamente, 75,8 kDa, determinada por SDS-PAGE, mas, em condições redutoras, apareceu como uma banda maior de 15,8 kDa e outra menor com 10,5 kDa. Sua sequência NH2-terminal revelou haver similaridade (89%) com o precursor putativo da albumina 2S de R. communis já descrita, com a cadeia A da estrutura da RicC3 (89%), com a cadeia A da mabilin-1, ambas, também, de R. communis (89%), com a cadeia pequena de uma proteina “napin-like”de Brassica napus (89%). Em todas essas proteínas similares, dois domínios, QEVQRKDLS e YLRQS, são altamente conservados. Comparação da estrutura primária gerada por ESI-Q-TOF MS/MS, a apartir de 17 peptídeos trípticos da Rc-2S-Alb, mostrou similaridade de 43% com a Mabinlin-1 (pI/Mr de 6,7 e 29.3 kDa) de R. communis. Em relação à estrutura tridimensional da Rc-2S-Alb, ela apresentou similaridade com a de Ric C3 e Mabinlin-1. A Rc-2S-Alb foi incapaz de inibir a germinação de esporos dos fungos fitopatogênicos Fusarium oxysporum e Rizoctonia solani, mas foi capaz de promover a aglomeração dos mesmos. A Rc-2S-Alb também não foi capaz de inibir o crescimento micelial dos fungos Fusarium oxysporum, Fusarium solani, Rizoctonia solani e Collethotricum gloeosporioides. Por outro lado, a Rc-2S-Alb foi eficiente em inibir o crescimento de Pseudomonas aeruginosae, Klebsiella pneumoniae e Bacillus subtilis, todas as bactérias patogênicas a seres humanos, quando em baixas concentrações. Como conclusão, foi possível a purificação de uma nova albumina 2S da torta da mamona, capaz de inibir tripsina e com atividade antibacteriana importante. Assim, a Rc-2S-Alb deve ser explorada no sentido de verificar sua eficácia como um novo agente terapêutico alternativo no combate a bactérias resistentes aos produtos disponíveis, hoje, no mercado, o que, se confirmado, poderá contribuir para a melhoria da saúde humana.

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Conselho Nacional de Desenvolvimento CientÃfico e TecnolÃgico / FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / Castor bean (Ricinus communis L.) is an important crop for the Northeast of Brazil, which recently has been used to produce biodiesel. Around 90% of the castor bean production in Brazil is concentrated at Northeast region and the state of Cearà is the second largest producer. During the oil extraction process from the castor bean seeds, a resulting residue, called castor cake, is underutilized. However, this byproduct is rich in protein and micronutrients such as nitrogen, phosphorus and potassium, an attribute that qualifies it as a stuff that could be used as organic fertilizer or as animal feed, for example, bringing added value to it. Unfortunately, its use as food has not been possible because of the presence of toxic elements and allergens (ricin, ricinine, complex allergens) in its composition, unless it were previously submitted to a detoxification process. To date, the existing technologies to this end are not economically viable on an industrial scale. Therefore, studies to add value to this abundant byproduct generated in the castor bean biodiesel productive chain is of paramount importance. Thus, in this context, this study was performed to identify, isolate, purify and characterize new bioactive molecules of de-oiled castor cake with biotechnological potential. Through extraction of soluble proteins with 50 mM Tris-HCl pH 7.5, fractionation with ammonium sulfate (50-75%), following by hydrophobic interaction chromatography in Phenyl-Sepharose column and ion exchange chromatography (DEAE-Sepharose), a protein, named Rc-2S-Alb, able to inhibit trypsin was purified. Rc-2S-Alb has a molecular mass of approximately 75.8 kDa, as determined by SDS-PAGE, and under reducing conditions showed a large and a small protein band of 15.8 kDa and 10.5 kDa, respectively. Its NH2-terminal sequence showed similarity with the following proteins: putative 2S albumin precursor (89%), chain A of RicC3 (89%), and chain A of mabilin-1 (89%), all from R. communis seeds; and with the short chain of a "napin-like" protein from Brassica napus (89%). In addition, these similar proteins have two highly conserved domains: QEVQRKDLS and YLRQS. Comparison of the partial primary structure of Rc-2S-Alb generated by the ESI-Q-TOF MS/MS analysis of 17 tryptic peptides, showed 43% similarity to Mabinlin-1 (pI/Mr 6.7 and 29.3 kDa) from R. communis. There were also high similarities among the three-dimensional structures of Rc-2S-Alb, RicC3 and Mabinlin-1. Rc-2S-Alb did not inhibit the spore germination of the phytopathogenic fungi Fusarium oxysporum and Rizoctonia solani, but promoted aggregation of their respective spores. Moreover, Rc-2S-Alb did not inhibit the mycelial growth of Fusarium oxysporum, Fusarium solani, Rizoctonia solani and Collethotricum gloeosporioides. Contrary, Rc-2S-Alb was effective in inhibiting the growth of the human pathogenic bacteria Pseudomonas aeruginosae, Klebsiella pneumoniae and Bacillus subtilis, at low concentrations. In conclusion, it was established a protocol to purify a new 2S albumin from castor bean cake, which inhibits trypsin and has important antibacterial activity. Thus, the Rc-2S-Alb should be further studied in order to verify its effectiveness as new alternative therapeutic agent against resistant bacteria to commercial antibiotics, which will contribute to improve the human health. / A mamoneira (Ricinus communis L.) à uma cultura importante para a regiÃo Nordeste do Brasil, onde, recentemente, tem sido utilizada para produÃÃo de biodiesel. O Nordeste detÃm 90% da produÃÃo brasileira de mamona, sendo o CearÃ, o segundo maior produtor. No processo de extraÃÃo de Ãleo das sementes de mamoneira para produÃÃo de biodiesel, o resÃduo resultante, denominado de torta da mamona, representa um subproduto pouco utilizado. Apesar disso, essa torta à rica em proteÃnas e micronutrientes, como nitrogÃnio, fÃsforo e potÃssio, atributo que a qualifica como um insumo que poderia ser utilizado como adubo orgÃnico, ou mesmo como raÃÃo animal, o que lhe agregaria valor comercial. Todavia, seu uso como alimento nÃo tem sido possÃvel por causa da presenÃa de elementos tÃxicos e alergÃnicos (Ricina, Ricinina, Complexos AlergÃnicos) na sua composiÃÃo, a nÃo ser que passasse por processamento para sua destoxificaÃÃo. Infelizmente, tecnologia economicamente viÃvel para esse fim, em escala industrial, ainda à inexistente. Portanto, hà necessidade de pesquisas que venham a agregar valor a esse subproduto abundante da cadeia produtiva do biodiesel. Assim, nesse contexto, o presente trabalho està inserido num projeto cujo objetivo maior à identificar, isolar, purificar e caracterizar novas molÃculas bioativas da torta delipidada de sementes de mamoeira com potencial biotecnolÃgico. AtravÃs de extraÃÃo de proteÃnas solÃveis com tampÃo Tris-HCl, 50 mM, pH 7,5, e fracionamento do extrato obtido com sulfato de amÃnio (50-75%), cromatografia de interaÃÃo hidrofÃbica em coluna Phenyl-Sepharose e cromatografia de troca iÃnica (DEAE-Sepharose) foi possÃvel purificar uma albumina 2S, denominada Rc-2S-Alb, capaz de inibir tripsina. A Rc-2S-Alb apresentou massa molecular de, aproximadamente, 75,8 kDa, determinada por SDS-PAGE, mas, em condiÃÃes redutoras, apareceu como uma banda maior de 15,8 kDa e outra menor com 10,5 kDa. Sua sequÃncia NH2-terminal revelou haver similaridade (89%) com o precursor putativo da albumina 2S de R. communis jà descrita, com a cadeia A da estrutura da RicC3 (89%), com a cadeia A da mabilin-1, ambas, tambÃm, de R. communis (89%), com a cadeia pequena de uma proteina ânapin-likeâde Brassica napus (89%). Em todas essas proteÃnas similares, dois domÃnios, QEVQRKDLS e YLRQS, sÃo altamente conservados. ComparaÃÃo da estrutura primÃria gerada por ESI-Q-TOF MS/MS, a apartir de 17 peptÃdeos trÃpticos da Rc-2S-Alb, mostrou similaridade de 43% com a Mabinlin-1 (pI/Mr de 6,7 e 29.3 kDa) de R. communis. Em relaÃÃo à estrutura tridimensional da Rc-2S-Alb, ela apresentou similaridade com a de Ric C3 e Mabinlin-1. A Rc-2S-Alb foi incapaz de inibir a germinaÃÃo de esporos dos fungos fitopatogÃnicos Fusarium oxysporum e Rizoctonia solani, mas foi capaz de promover a aglomeraÃÃo dos mesmos. A Rc-2S-Alb tambÃm nÃo foi capaz de inibir o crescimento micelial dos fungos Fusarium oxysporum, Fusarium solani, Rizoctonia solani e Collethotricum gloeosporioides. Por outro lado, a Rc-2S-Alb foi eficiente em inibir o crescimento de Pseudomonas aeruginosae, Klebsiella pneumoniae e Bacillus subtilis, todas as bactÃrias patogÃnicas a seres humanos, quando em baixas concentraÃÃes. Como conclusÃo, foi possÃvel a purificaÃÃo de uma nova albumina 2S da torta da mamona, capaz de inibir tripsina e com atividade antibacteriana importante. Assim, a Rc-2S-Alb deve ser explorada no sentido de verificar sua eficÃcia como um novo agente terapÃutico alternativo no combate a bactÃrias resistentes aos produtos disponÃveis, hoje, no mercado, o que, se confirmado, poderà contribuir para a melhoria da saÃde humana.

La chine subtropicale est considérée comme une zone riche en biodiversité ainsi qu'une réserve naturelle abritant plusieurs espèces de plantes endémiques sous l'influence de la Mousson Asiatique (MA). Les enregistrements paléoclimatiques continus dans cette région sont trop peu nombreux pour comprendre l'évolution floristique liée aux changements climatiques qui demeure ainsi méconnue, particulièrement durant le DMG. Dans ce contexte, ce travail a pour objectifs d'étudier, au cours des derniers 40000 ans, la variabilité floristique et climatique enregistrées dans les zones humides montagneuse sub-tropicales chinoises. Notre étude s'appuie sur l'étude de deux carottes sédimentaires ou la variabilité est reconstituée à partir de l'étude palynologique. L'enregistrement le plus long (derniers 42000 ans) a été obtenu sur une carotte prélevée dans la zone humide sub-alpine de Dajiuhu, dans les montagnes Shennongjia situées en Chine centrale. Cette région est fortement influencée par la Mousson Est Asiatique (MEA) et caractérisée par des forets tempérées décidues associées à quelques taxons de conifères. L'autre enregistrement concerne une carotte (GT-2), qui quant à elle enregistre les derniers 21000 ans, prélevée dans la zone humide de Gutian, province de Guangxi au sud de la chine sous l'influence de la mousson indienne (MI). L'objectif de ce travail est de reconstituer les variations régionales de la végétation et les variations climatiques liées aux deux systèmes de mousson (MEA et MI). L'étude palynologique de nos deux carottes est complétée et renforcée par une approche multi-proxy s'appuyant sur les spores, le δ13C, l'analyse de l'échelle de gris, la susceptibilité magnétique ainsi que le degré d'humification. L'analyse du pollen dans la carotte DJH-1 révèle que le climat et la végétation ont significativement varié tout au long de ces derniers 42000 ans. La région de Dajiuhu, caractérisée aujourd'hui par des forêts denses était autrefois couvertes par des prairies alpine associés à une forêt clairsemée durant le DMG comme le laisse suggérer la prédominance des Cyperaceae et Poaceae dans les spectres polliniques. De faibles teneurs en matière organique (MOT) ainsi qu'un niveau de gris bas ont été aussi signalés durant la période glaciaire. La foret commençait à se reconstituer à partir de 14ka BP et les forets à arbre persistant ont atteint leur niveau maximale entre 10ka et 4ka BP, correspondant au maximum thermique de l'holocène. Les changements survenus après 4000BP ont été explorés aussi. La carotte GT-2 au sud de la chine sub-tropicale révèle quant à elle une végétation différente durant le dernier maximum glaciaire. Les spectres polliniques indiquent que cette période était caractérisée par des forêts de conifères associées à des forêts décidues. La prépondérance des genres décidus comme Carpinus, Betula et Corylus semble indiquer des conditions froides, alors que les pluies restaient abondantes. L'abondance du genre Tsuga, particulièrement entre 21 et 17ka BP suggère la descente de la limite forestière à des altitudes beaucoup plus basses. Les successions de biomes dans montagnes subtropicale du sud de la chine s'est étalée sur plusieurs phases : (1) la première se caractérise par des forêts décidues associées à des conifères (17-12.5ka BP), (2) la deuxième phase est dominée par les forêts decidues, (3) la troisième se caractérise par des forêts à arbres persistants associées à des arbres a feuilles caduques (12.5-9ka BP). Les deux dernières phases voient se succéder des forêts à arbres persistants (9-2.5ka BP) puis une forêt mixte, probablement liée à l'activité humaine. La comparaison entre les deux sites d'étude confirme que les changements de végétation durant le DMG étaient importants. / Subtropical China is considered as an important region for biodiversity and a great natural reserve for endemic plant species, where the climate is mainly controlled by Asian Monsoon (AM). Since the recent decades, few continuous records so far covering the last glacial period in this area have been studied. In consequence, the past floristic evolution and vegetation-climate changes during particularly the Last Glacial period are still unclear. In this study, two cores from subtropical mountain wetlands were studied by means of palynology and other multidisciplinary proxies. The longer studied material covering the last 42 ka was obtained from a sub-alpine wetland in Dajiuhu, Shennongjia Mountains of central China, where the present-day vegetation is temperate deciduous forest mixed with some conifer taxa, and the climate is greatly influenced by the East Asian monsoon (EAM). The other material, core GT-2, collected in Gutian wetland of Guangxi Province in south China has an age of 21 ka can reflect the changes of evergreen forest and climate which is controled by the Indian monsoon (IM) overlapped with EAM. The aim of the current study is to reconstruct the regional vegetation and climate changes, and better understand the variability of two monsoon system (EAM and IM) since the last glacial period. Multi-proxy analysis including pollen and spore, stable carbon isotope (δ13C), sediment gray-scale (GS), magnetic suscepbitility (MS),peat humification (HD) and so on were perfomed for evaluating the regional environment changes. The overall result can be summarized as below: The pollen analysis from the DJH-1 core reveals that the past vegetation and climate in northern subtropical zone of central China varied significantly over the last 42 ka. The vegetation in Dajiuhu region, dominated by dense temperate forest today, was an alpine meadow with sparse mixed forest during the last glacial characterized by predominant Cyperaceae and Poaceae in the pollen spectra. Other proxies show that the lowest TOC content and lighter gray-scale in the glacial interval. The broadleaved forest began to return since 14 cal ka BP, and the evergreen broadleaved trees attained their highest level between ~10 and 4 cal ka BP, accordance in timing with the Holocene thermal maximum. A change at 4000 cal BP in pollen spretra was also investigated. The results from the core GT-2 in southern subtropical zone of China suggest a different replacement of vegetation during the last glacial. Pollen data indicate that a dense mixed forest of coniferous and deciduous broadleaved forest covered the southern subtropical mountains during the LGM (21-12.5 cal ka BP). The grass was however in low percentage. The high percentage of deciduous taxa such as Carpinus, Betula and Corylus indicate a colder condition, whereas the rainfall maintains abundant. The relatively high amount of Tsuga particularly during 21-17 cal ka BP suggests an important lowering of vertical forest belt. The biome changes in the southern subtropical mountains since the last glacial maximum can be outlined as follow: (1) deciduous and coniferous mixed forest (21-17 ka BP); (2) deciduous broadleaved forest (17-12.5 ka BP); (3) deciduous and evergreen mixed forest (12.5-9 ka BP); (4) evergreen broadleaved forest (9-2.5 cal ka BP) and (5) Mixed forest (from 2.5 cal ka BP) possible caused by human activity. The comparison of the two studied sites confirms that the shift of vegetation zone during the last glacial period is important. The alpine tree line might decend at an amplitude of more than 1000 m lower than that of today, and the temperate zone of deciduous broadleaved forest moved southwards to Guangxi Province (e.g. from about 30 to 22 latitude N ). The rapid increase of braodleved forest in central China began at about 10 cal ka BP, whereas the return to evergreen forest in Guangxi of southern subtropical zone took place at ca. 9 cal ka BP.

Anthrax spores, in weaponized form, are dangerous biological warfare agents. Handheld technology for the rapid detection of anthrax is greatly needed to improve national security. Methods to detect anthrax spores are diverse, with most taking at least an hour for positive identification. A viable option for rapid detection is analytical pyrolysis (AP), which produces chemicals containing taxonomical information (biomarkers). AP methods are reviewed and critically analyzed to show that reproducible detection of anthrax spores in a rapid manner (< 5 min) with a handheld device is not currently possible. A promising alternative to AP is the use of a catalyst to produce biomarkers from anthrax spores with improved selectivity and reproducibility. Catalytic materials having promise for this include platinum, nickel, and superacids. Experiments evaluating several of these materials are described. A biomarker mass spectral library was created, based on information available in the scientific literature, to facilitate analysis and identification of the biomarkers produced experimentally. The RAMFAC algorithm was used to deconvolute chromatographic peaks to produce clean mass spectra and match them against entries in the biomarker library. While the library is not complete, its use with the RAMFAC algorithm enabled detection of many important biomarkers in experiments involving catalytic breakdown of anthrax spores. Experimental results from preliminary tests of several catalysts are presented and discussed. Addition of catalysts in the form of platinum nanoclusters and superacids to bacterial spores in a commercial pyrolyzer effected an increase in the amount of biomarkers produced at mild conditions over traditional pyrolysis methods. Electroformed nickel mesh, on the other hand, demonstrated low catalytic activity for the production of biomarkers, likely due to poor contact of the spores with the mesh. Biomarkers similar to those published in the literature were observed, including dipicolinic acid, picolinic acid, propionamide, acetamide, diketopiperazines, fatty acids, furfuryl alcohol, and DNA bases. A statistically designed factorial study was used to determine the importance of temperature, spore loading, and nanocluster loading on the production of three important biomarkers. The relative importance of these variables differs for each of the three important biomarkers, suggesting they are produced by different reaction mechanisms.