This design project explores the complexity of witchcraft, focusing on the socio-cultural and ecological levels. The project’s transdisciplinarity bridges the historical facts of witchcraft, folktales, culture, and natural science practice with speculative design. It studies how, in the past, witchcraft was used against women as a patriarchal system of oppression and the ways in which it has been reclaimed through feminism in the present. It also looks into how women’s scientific participation and contributions have been partly and entirely erased or wrongly credited to men. Through practice-based research and multispecies collaboration, this project aims to empower women to learn and experiment with natural science through an ecofeminist lens and framework. The project offers a speculative design exploration of how today's witches can contribute to environmental restorative practices, through a communicational tool with the incredible mycorrhizal network found underground. This is directed toward young women who represent the future of environmental science and justice.

Sclerotinia blight, caused by <i>Sclerotinia minor</i>, is a severe disease of peanut in Virginia. Vinclozolin (V), iprodione (I), dicloran (D), and pentachloronitrobenzene (PCNB) were evaluated for their fungitoxicity to <i>S. minor</i>. The mean ED₅₈ values for five isolates were found to be 0.07, 0.11, 0.91, and 1.27 μg/ml, for V, I, D, and PCNB, respectively, on fungicide-amended glucose yeast-extract agar (GYEA). Fungicide-resistant growth sectors developed on media amended with I or V. Nine such strains occurred; they were capable of growth on GYEA amended with up to 1000 μg/ml of I or V, and were cross-resistant to D or PCNB. Resistance was maintained in all but two strains after repeated culture in the absence of fungicide for 3 yr. In field microplots, two resistant strains were pathogenic to peanut and survived as well as a fungicide-sensitive field isolate. D, I and V were applied to peanuts in the microplots for 3 yr at total annual rates of 8.41, 3.36, and 2.52 kg/ha, respectively. Disease severity caused by the resistant strains was suppressed 19, 33, and 87% by D, I, and V, respectively, as compared to 15, 24, and 76% for the sensitive isolate. Isolates recovered from tissue biopsies still grew on fungicide-amended GYEA indicating that <i>in vitro</i> and <i>in vivo</i> resistance are not equivalent in this case. Fungicide treatments reduced sclerotial populations of all strains, and reduced the viability of sclerotia from sensitive but not resistant strains. Fungicide-resistant strains were capable of surviving and competing pathogenically in microplots infested with equal numbers of sclerotia from a sensitive and a resistant strain; this trend was enhanced by fungicide applications. A survey of 763 isolates from fields treated with these fungicides failed to detect resistant strains. One fungicide-resistant isolate was recovered from an iprodione-treated microplot originally infested with a sensitive field isolate. A technique utilizing excised peanut stems was devised to evaluate isolate pathogenicity, cultivar resistance to the disease, susceptibility of different age peanut tissues, and fungicide persistence on peanut stems in the field. The method was also used to screen fungicides; results verified previous findings which indicated that <i>in vitro</i> resistance is not equivalent to <i>in vivo</i> resistance. Resistance to these fungicides may eventually become a field problem, but with correct management they should provide years of disease control. / Ph. D. / incomplete_metadata

A paracoccidioidomicose (PCM) é uma doença granulomatosa sistêmica, causada por Paracoccidioides spp., (P. brasiliensis e P. lutzii), geograficamente, limita-se a América Latina com as áreas endêmicas estendendo-se desde o México até a Argentina, constituindo uma das micoses sistêmicas de maior incidência na região, afetando principalmente trabalhadores rurais. O maior número de pacientes com PCM tem sido reportado principalmente no Brasil, Colômbia e Venezuela. A incidência real desta micose encontra-se subestimada no Brasil e pouco se conhece em relação a nova espécie descrita - P. lutzii. A maioria dos estudos em P. lutzii foram focados em genética, especiação e na geração de novos antígenos para melhorar a especificidade e sensibilidade dos testes sorológicos. Atualmente, as preparações antigênicas tradicionais, preparadas a partir de isolados de P. brasiliensis, são ineficientes. Raros são os trabalhos focados na biologia de P. lutzii e nos fatores de virulência que podem ser comparados com P. brasiliensis nos modelos experimentais. A nossa proposta de estudo foi avaliar alguns aspectos in vitro e in vivo relacionados com a patogenicidade e destacamos: a fagocitose e a morte intracelular de P. lutzii por macrófagos, peritoneais, de camundongos Knockouts (KO) e selvagens para PRRs (TLR2, TLR4 e Dectina) e ativadores intracelulares (MyD88 e NALP3). Paralelamente a este estudo, animais foram infectados com leveduras de P. lutzii e comparados com os modelos de infecção já estabelecidos com leveduras (Pb18) e conídios (ATCCPb60855) de P. brasiliensis. Nossos dados indicam que similar ao que ocorre com P. brasiliensis a fagocitose de P. lutzii depende de TLR2, TLR4 e Dectina- 1, resultados semelhantes também foram observadas na expressão de moléculas envolvidas na co-estimulação e a apresentação de antígenos (MHC II, CD80 e CD86). Contudo, a morte intracelular de leveduras de P. lutzii é claramente dependente de TLR4, e a produção de citocinas IL-6, MIP-2, IFN- e IL-12p40 são importantes para o controle das leveduras pelos macrófagos. No modelo experimental de P. lutzii, camundongos machos C57BL/6 (6-7 semanas) foram infectados intratraquealmente como 1x106 leveduras viáveis do isolado de P. lutzii Pb01. Encontramos duas fases da doença, a primeira de 0 hora até 2 a 4 semanas pós-infecção, e a segunda de 4 até 12 semanas. As leveduras parecem ser contidas na primeira semana de infecção e posteriormente não encontramos leveduras nos macerados de pulmão, diferente do modelo de BALB/c infetado com conídios de ATCC-Pb60855 no qual as UFC são recuperadas até a semana 16 pós-infeção. Como relação aos níveis de citocinas, encontramos que na lavagem broncoalveolar e macerado de pulmão um perfil misto Th1/Th2 porém, marcado por citocinas próinflamatórias no primeiro período e citocinas regulatórias tipo Th2 no segundo período (IL-12p70, IL-23, IL-10); similar ao descrito nos modelos de P. brasiliensis infectados tanto com conídios como com leveduras. No entanto, no primeiro período da doença, em camundongos C57BL/6, parece ter uma carga inflamatória maior que reflete nas citocinas que mantém seus níveis até o período crônico: TNF-alfa, MIP-2 e GM-CSF está última, regulada positivamente tanto em experimentos in vitro como in vivo. Também observamos que a partir das 48horas pós-infecção encontramos níveis aumentados de IL-12p70 até o período crônico onde junto com a IL-23 parecem ser as responsáveis pela diminuição da infecção no período tárdio. Esta é a primeira vez que se descreve um modelo experimental com P. lutzii (isolado Pb01) indicando o perfil imunopatológico com pequenas diferenças comparados ao P. brasiliensis porém, de importância na patogenicidade da doença auxiliando a compreender as diferentes formas da doença no modelo experimental / Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides spp. (P. brasiliensis and P. lutzii), geographically, is limited to Latin America with endemic areas from Mexico to Argentina, as one of the systemic mycoses with the highest incidence in the region, mainly affecting rural workers. The largest number of patients with PCM has been mainly reported in Brazil, Colombia and Venezuela. The true incidence of this mycosis is underestimated in Brazil and little is known about the new species described - P. lutzii. Most studies in P. lutzii were focused on genetics, speciation and the generation of new antigens to improve the specificity and sensitivity of serological tests. Currently, traditional antigenic preparations, prepared with isolates of P. brasiliensis, are inefficient. There are few studies focused on P. lutzii biology and virulence factors that can be compared with P. brasiliensis in experimental models. Our study aimed to evaluate some in vitro and in vivo aspects related to pathogenicity: phagocytosis and intracellular killing of P. lutzii by peritoneal macrophages from knockouts (KO) for PRRs (TLR2, TLR4 and Dectin) and intracellular activators (MyD88 and NALP3). In addition, animals were infected with P. lutzii yeast and compared with the well-established models of infection with yeast cells (Pb18) and conidia (ATCC Pb60855) from P. brasiliensis. Our data indicate that similarly to what happens with the phagocytosis of P. brasiliensis, P. lutzii phagocytosis is dependent on TLR2, TLR4 and Dectin-1. Other molecules, involved in co-stimulation and presentation of antigens such as MHC II, CD80 and CD86 were also shown to participate in the P. lutzii-host interaction. However, intracellular killing of P. lutzii yeast cells was clearly dependent on TLR4, and the production of cytokines as IL-6, MIP-2, IFN- and IL-12p40 were important for the control of the yeast by macrophages. In the experimental model of P. lutzii, male C57BL/6 mice (6-7 weeks) were infected intratracheally with 1x106 viable yeasts of the isolate Pb01like. We found two phases of the disease, the first from the inoculation to 2 or 4 weeks after infection and the second from 4 to 12 weeks. Yeast appear to be contained within the first week of infection and subsequently are also absent from macerated lung, differently from the model of BALB/c mice infected with ATCC Pb60855 conidia in which CFUs were detected up to week 16 post-infection. We found a mixed Th1/Th2 pattern (IL-12p70, IL-23, IL-10) in bronchoalveolar lavage and lung, with the predominance of proinflammatory cytokines in the first phase and predominance of regulatory Th2 cytokines in the second phase, reproducing findings of P. brasiliensis infection models produced with both conidia and yeast. However, in the first period of the disease in C57BL/6 mice there was a higher inflammatory burden, reflected by the high cytokine levels (TNF-alpha, MIP-2 and GM-CSF), the latter in particular because it was positively regulated both in vitro and vivo), that persisted through the chronic period. We also observed that starting from 48 hours postinfection to the chronic period there were increased levels of IL-12p70, which together with IL-23 appeared to be responsible for the reduction of infection in the late period. This is the first time that an experimental model with P. lutzii (Pb01) is described, showing an immunological profile with only slight differences compared to the P. brasiliensis model. The present study details important aspects of the pathogenesis of the disease due to different species of Paracoccidioides and helps to understand the different forms of presentation in experimental models

A 24-hour fermentation of the optimized synbiotic composed of B. longum and EPR was performed to give a cell-free fermentation broth (S24). S24 was co-cultured with two colon cancer cell lines (Caco-2 and SW620) and normal colon cells (FHC). S24 significantly inhibited cell proliferation for both colon cancer cells but promoted FHC cell growth by 10-25% as shown by MTT and BrdU arrays. Primary DNA damage analysis by alkaline comet assay showed S24 caused DNA damage to a comparable extent as the positive control of 10 mM H2O2 (treated for 1 hour) for both cancer cells. Dynamic analysis on DNA damage-associated DNA repair showed the two colon cancer cells had different response pattern to S24. Flow cytometric analysis showed that both Caco-2 and SW620 when treated with S24 (IC 50=3.66 mM of acetate) were arrested initially at G2/M and subsequently at S phase accompanied with large sub-G1 peaks. Dual staining with PI/AnnexinV further proved the appearance of apoptosis. Live cell imaging analysis on Caco-2 cells treated with S24 showed the following events: mitochondria were rapidly destroyed within the first two-hour treatment, the cells bubbled and the nucleus condensed after the mitochondrial had shrunken, followed by apoptosis. / Despite active research on synbiotic on anti-carcinogenesis of colon cancer by synbiotics, the underlying mechanism still remains unclear. This study investigated a novel synbiotic composed of non-digestible carbohydrates (NDCs) extracted from mushroom sclerotia as prebiotics and Bifidobacteria as probiotics. Preliminary results on incubation of two probiotics ( Bifidobacterium longum and Lactobacillus brevis) and one pathogenic bacterium (Clostridium celatum) separately with 3 NDCs extracted from mushroom sclerotia [Poria cocos (PC), Polyporus rhinocerus (PR) and Pleurotus tuber-regium (PT)] indicated that the growth of B. longum and L. brevis was stimulated more preferentially than C. celatum after 72-hour fermentation. The short-chain fatty acid (SCFA) profile was dominated by acetate (> 98% of total SCFAs) with very little butyrate (&lt; 2.0% of total SCFAs) and the organic matter disappearance (OMD) during fermentation was consistent with the bacterial growth. Among the synbiotic combinations, NDC from PR and B. longum gave the largest amount of acetate (2.47+/-0.232 mmol/g of organic matter disappearance). / Results obtained from human pathway finder RT2 Profiler(TM) PCR Array indicated that S24 could modulate the proliferation of colon cancer cells mainly by various pathways such as cell cycle and DNA damage repair, apoptosis and cell senescence, etc. In SW620 cells, PCR Array of Human Cell Cycle further revealed that the modulated genes mainly belonged to the gene cluster of S phase and DNA replication as well as G2 and G2/M transition. While for Caco-2 cells, the cell-cycle modulated genes mainly belonged to the cluster of G2 and G2/M transition. Immuno-blotting on the pivotal upstream regulators showed that phosphorylation of ATM at Serine 1981 was significantly increased in both cancer cells. Site-specific phosphorylation of pRB was decreased and phosphorylation of Chk1 was increased in both cancer cells while Chk2 were increased in SW620 cells. Cdc25A was phosphorylated at serine17 in both cancer cells. It can be proposed that the blockage of DNA synthesis or DNA damage was due to the down-regulation of some pivotal DNA replication related proteins such as RPA3, PCNA and MCMs, detected by ATM-Chk1/Chk2-Cdc25A pathway. This would cause the prolonged staying of cells at the G1/S checkpoint which further moved on to S phase arrest for SW620 cells. Moreover the sharply up-regulated p21, an important inhibitor of Cdk2 would further hinder the cells passing the G1/S checkpoint in SW620 cells. / The tumor suppressor p53 was detected phosphorylated at various sites in SW620 but not in Caco-2 cells. In SW620 cells, G2/M arrest was caused by the inhibition of CDK1/CDC2 due to increased expression of GADD45A and p21 and phosphorylation of Cdc25A, while for Caco-2, the G2/M arrest was caused by degradation of Cdc25A due to the absence of p53-activated GADD45A and p21 expression as shown in the pathway finder results. Some apoptosis-related proteins of Bax, Apaf-1 and PARP were modulated as shown by immuno-blotting in both colon cancer cells. (Abstract shortened by UMI.) / Gao, Shane. / Adviser: Peter Chi-Keung Cheung. / Source: Dissertation Abstracts International, Volume: 72-11, Section: B, page: . / Thesis (Ph.D.)--Chinese University of Hong Kong, 2009. / Includes bibliographical references (leaves 55-94). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Electronic reproduction. [Ann Arbor, MI] : ProQuest Information and Learning, [201-] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniversitet, 2006. / Härtill 4 uppsatser.

Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2006. / Härtill 4 uppsatser.

Ganoderma lucidum é um cogumelo medicinal consumido tradicionalmente nos países asiáticos que apresenta diversos efeitos benéficos comprovados. Apesar de todos os estudos sobre seus compostos bioativos, a melhor forma de enriquecimento de meios de cultivo visando aumentar a produção destes compostos ainda se mantém incerta. Além disso, poucos trabalhos tratam dos efeitos sobre o desempenho de animais de produção. Para testar diferentes meios de cultivo de micélio de G. lucidum, foram testados resíduos agrícolas (fermentação em estado sólido) e diferentes açúcares e aminoácidos (em meios de cultura com ágar), para avaliar o crescimento do micélio de G. lucidum. O fornecimento de farinha com micélio de G. lucidum obtido por fermentação em meio sólido (grãos de trigo) para coelhos também foi avaliado. O micélio de G. lucidum desenvolveu-se bem nos resíduos agrícolas, destacando-se a casquinha de soja que apresentou maior taxa de crescimento e também maior concentração de β-glucanas. No experimento em placas, o G. lucidum também se desenvolveu bem, destacando-se os meios que continham celobiose e tirosina, que apesar de apresentarem menores taxas de crescimento e crescimento total em 10 dias, produziram micélios com maior concentração de β-glucanas e capacidade antioxidante em equivalentes trolox (TEAC), respectivamente. Os coelhos não apresentaram sinais de intolerância ao consumo de ração com diferentes concentrações farinha com micélio de G. lucidum. Todos os parâmetros de desempenho e avaliação de carcaça foram adequados à idade em que foram abatidos. A avaliação histológica dos órgãos apresentou alterações nas células renais (tubulares e glomerulares), apontando para uma possível lesão renal conforme aumentou a concentração de farinha com micélio na ração. A avaliação histomorfométrica resultou em maior altura de vilosidade de íleo e menor largura de vilosidade de jejuno na concentração 0,5%, e redução do diâmetro de cripta conforme o aumento da concentração de farinha com micélio. Estes resultados indicam a possibilidade de estudos mais aprofundados sobre os aspectos de utilização de celobiose e tirosina para produção de compostos bioativos, e sobre aspectos de toxicidade do micélio deste cogumelo para segurança no fornecimento para animais. / Ganoderma lucidum is a medicinal mushroom traditionally consumed in Asian countries that presents several beneficial effects already verified. Despite all studies about their bioactive compounds, the best cultivation media enrichment aiming to increase the production of these compounds is still uncertain. Besides, few studies are related to the performance of production animals. In order to test different cultivation media for G. lucidum mycelia, agricultural residues (solid state fermentation) and different sugars and aminoacids (in culture media with agar), were tested to evaluate G. lucidum mycelium growth. Supply of flour with G. lucidum mycelia obtained by solid state fermentation (wheat grain) for rabbits was also evaluated. Mycelium of G. lucidum developed very well in all agricultural residues, soybean hulls was the residue that presented higher growth rate and higher concentration of β-glucans. In the cultivation media experiment, G. lucidum also developed well, media that contained cellobiose and tyrosine, despite presenting lower growth rates and total growth within 10 days, produced mycelia with higher concentration of β-glucans and trolox equivalent antioxidant capacity (TEAC), respectively. Rabbits did not show any sign of intolerance to feed with different concentrations of flour with G. lucidum mycelium. All performance parameters and dressing percentages were adequate to the age at which they were slaughtered. Histological evaluation of organs presented alterations in renal cells (tubular and glomerulus), indicating a possible renal lesion according to the increase of flour with mycelium in feed. Histomorphometric evaluation showed increased vilous height in ileum and decreased vilous width of jejunus at 0.5% concentration, and decrease in crypt diameter according to the increase of concentration of flour with mycelium in feed. These results indicate the possibility of more studies regarding the aspects about cellobiose and tyrosine utilization for the production of bioactive compounds, and about toxicity of this mushroom mycelia, assuring the safety in supplying this product for animails.

Ganoderma lucidum é um cogumelo medicinal consumido tradicionalmente nos países asiáticos que apresenta diversos efeitos benéficos comprovados. Apesar de todos os estudos sobre seus compostos bioativos, a melhor forma de enriquecimento de meios de cultivo visando aumentar a produção destes compostos ainda se mantém incerta. Além disso, poucos trabalhos tratam dos efeitos sobre o desempenho de animais de produção. Para testar diferentes meios de cultivo de micélio de G. lucidum, foram testados resíduos agrícolas (fermentação em estado sólido) e diferentes açúcares e aminoácidos (em meios de cultura com ágar), para avaliar o crescimento do micélio de G. lucidum. O fornecimento de farinha com micélio de G. lucidum obtido por fermentação em meio sólido (grãos de trigo) para coelhos também foi avaliado. O micélio de G. lucidum desenvolveu-se bem nos resíduos agrícolas, destacando-se a casquinha de soja que apresentou maior taxa de crescimento e também maior concentração de β-glucanas. No experimento em placas, o G. lucidum também se desenvolveu bem, destacando-se os meios que continham celobiose e tirosina, que apesar de apresentarem menores taxas de crescimento e crescimento total em 10 dias, produziram micélios com maior concentração de β-glucanas e capacidade antioxidante em equivalentes trolox (TEAC), respectivamente. Os coelhos não apresentaram sinais de intolerância ao consumo de ração com diferentes concentrações farinha com micélio de G. lucidum. Todos os parâmetros de desempenho e avaliação de carcaça foram adequados à idade em que foram abatidos. A avaliação histológica dos órgãos apresentou alterações nas células renais (tubulares e glomerulares), apontando para uma possível lesão renal conforme aumentou a concentração de farinha com micélio na ração. A avaliação histomorfométrica resultou em maior altura de vilosidade de íleo e menor largura de vilosidade de jejuno na concentração 0,5%, e redução do diâmetro de cripta conforme o aumento da concentração de farinha com micélio. Estes resultados indicam a possibilidade de estudos mais aprofundados sobre os aspectos de utilização de celobiose e tirosina para produção de compostos bioativos, e sobre aspectos de toxicidade do micélio deste cogumelo para segurança no fornecimento para animais. / Ganoderma lucidum is a medicinal mushroom traditionally consumed in Asian countries that presents several beneficial effects already verified. Despite all studies about their bioactive compounds, the best cultivation media enrichment aiming to increase the production of these compounds is still uncertain. Besides, few studies are related to the performance of production animals. In order to test different cultivation media for G. lucidum mycelia, agricultural residues (solid state fermentation) and different sugars and aminoacids (in culture media with agar), were tested to evaluate G. lucidum mycelium growth. Supply of flour with G. lucidum mycelia obtained by solid state fermentation (wheat grain) for rabbits was also evaluated. Mycelium of G. lucidum developed very well in all agricultural residues, soybean hulls was the residue that presented higher growth rate and higher concentration of β-glucans. In the cultivation media experiment, G. lucidum also developed well, media that contained cellobiose and tyrosine, despite presenting lower growth rates and total growth within 10 days, produced mycelia with higher concentration of β-glucans and trolox equivalent antioxidant capacity (TEAC), respectively. Rabbits did not show any sign of intolerance to feed with different concentrations of flour with G. lucidum mycelium. All performance parameters and dressing percentages were adequate to the age at which they were slaughtered. Histological evaluation of organs presented alterations in renal cells (tubular and glomerulus), indicating a possible renal lesion according to the increase of flour with mycelium in feed. Histomorphometric evaluation showed increased vilous height in ileum and decreased vilous width of jejunus at 0.5% concentration, and decrease in crypt diameter according to the increase of concentration of flour with mycelium in feed. These results indicate the possibility of more studies regarding the aspects about cellobiose and tyrosine utilization for the production of bioactive compounds, and about toxicity of this mushroom mycelia, assuring the safety in supplying this product for animails.

A paracoccidioidomicose (PCM) é uma doença granulomatosa sistêmica, causada por Paracoccidioides spp., (P. brasiliensis e P. lutzii), geograficamente, limita-se a América Latina com as áreas endêmicas estendendo-se desde o México até a Argentina, constituindo uma das micoses sistêmicas de maior incidência na região, afetando principalmente trabalhadores rurais. O maior número de pacientes com PCM tem sido reportado principalmente no Brasil, Colômbia e Venezuela. A incidência real desta micose encontra-se subestimada no Brasil e pouco se conhece em relação a nova espécie descrita - P. lutzii. A maioria dos estudos em P. lutzii foram focados em genética, especiação e na geração de novos antígenos para melhorar a especificidade e sensibilidade dos testes sorológicos. Atualmente, as preparações antigênicas tradicionais, preparadas a partir de isolados de P. brasiliensis, são ineficientes. Raros são os trabalhos focados na biologia de P. lutzii e nos fatores de virulência que podem ser comparados com P. brasiliensis nos modelos experimentais. A nossa proposta de estudo foi avaliar alguns aspectos in vitro e in vivo relacionados com a patogenicidade e destacamos: a fagocitose e a morte intracelular de P. lutzii por macrófagos, peritoneais, de camundongos Knockouts (KO) e selvagens para PRRs (TLR2, TLR4 e Dectina) e ativadores intracelulares (MyD88 e NALP3). Paralelamente a este estudo, animais foram infectados com leveduras de P. lutzii e comparados com os modelos de infecção já estabelecidos com leveduras (Pb18) e conídios (ATCCPb60855) de P. brasiliensis. Nossos dados indicam que similar ao que ocorre com P. brasiliensis a fagocitose de P. lutzii depende de TLR2, TLR4 e Dectina- 1, resultados semelhantes também foram observadas na expressão de moléculas envolvidas na co-estimulação e a apresentação de antígenos (MHC II, CD80 e CD86). Contudo, a morte intracelular de leveduras de P. lutzii é claramente dependente de TLR4, e a produção de citocinas IL-6, MIP-2, IFN- e IL-12p40 são importantes para o controle das leveduras pelos macrófagos. No modelo experimental de P. lutzii, camundongos machos C57BL/6 (6-7 semanas) foram infectados intratraquealmente como 1x106 leveduras viáveis do isolado de P. lutzii Pb01. Encontramos duas fases da doença, a primeira de 0 hora até 2 a 4 semanas pós-infecção, e a segunda de 4 até 12 semanas. As leveduras parecem ser contidas na primeira semana de infecção e posteriormente não encontramos leveduras nos macerados de pulmão, diferente do modelo de BALB/c infetado com conídios de ATCC-Pb60855 no qual as UFC são recuperadas até a semana 16 pós-infeção. Como relação aos níveis de citocinas, encontramos que na lavagem broncoalveolar e macerado de pulmão um perfil misto Th1/Th2 porém, marcado por citocinas próinflamatórias no primeiro período e citocinas regulatórias tipo Th2 no segundo período (IL-12p70, IL-23, IL-10); similar ao descrito nos modelos de P. brasiliensis infectados tanto com conídios como com leveduras. No entanto, no primeiro período da doença, em camundongos C57BL/6, parece ter uma carga inflamatória maior que reflete nas citocinas que mantém seus níveis até o período crônico: TNF-alfa, MIP-2 e GM-CSF está última, regulada positivamente tanto em experimentos in vitro como in vivo. Também observamos que a partir das 48horas pós-infecção encontramos níveis aumentados de IL-12p70 até o período crônico onde junto com a IL-23 parecem ser as responsáveis pela diminuição da infecção no período tárdio. Esta é a primeira vez que se descreve um modelo experimental com P. lutzii (isolado Pb01) indicando o perfil imunopatológico com pequenas diferenças comparados ao P. brasiliensis porém, de importância na patogenicidade da doença auxiliando a compreender as diferentes formas da doença no modelo experimental / Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by Paracoccidioides spp. (P. brasiliensis and P. lutzii), geographically, is limited to Latin America with endemic areas from Mexico to Argentina, as one of the systemic mycoses with the highest incidence in the region, mainly affecting rural workers. The largest number of patients with PCM has been mainly reported in Brazil, Colombia and Venezuela. The true incidence of this mycosis is underestimated in Brazil and little is known about the new species described - P. lutzii. Most studies in P. lutzii were focused on genetics, speciation and the generation of new antigens to improve the specificity and sensitivity of serological tests. Currently, traditional antigenic preparations, prepared with isolates of P. brasiliensis, are inefficient. There are few studies focused on P. lutzii biology and virulence factors that can be compared with P. brasiliensis in experimental models. Our study aimed to evaluate some in vitro and in vivo aspects related to pathogenicity: phagocytosis and intracellular killing of P. lutzii by peritoneal macrophages from knockouts (KO) for PRRs (TLR2, TLR4 and Dectin) and intracellular activators (MyD88 and NALP3). In addition, animals were infected with P. lutzii yeast and compared with the well-established models of infection with yeast cells (Pb18) and conidia (ATCC Pb60855) from P. brasiliensis. Our data indicate that similarly to what happens with the phagocytosis of P. brasiliensis, P. lutzii phagocytosis is dependent on TLR2, TLR4 and Dectin-1. Other molecules, involved in co-stimulation and presentation of antigens such as MHC II, CD80 and CD86 were also shown to participate in the P. lutzii-host interaction. However, intracellular killing of P. lutzii yeast cells was clearly dependent on TLR4, and the production of cytokines as IL-6, MIP-2, IFN- and IL-12p40 were important for the control of the yeast by macrophages. In the experimental model of P. lutzii, male C57BL/6 mice (6-7 weeks) were infected intratracheally with 1x106 viable yeasts of the isolate Pb01like. We found two phases of the disease, the first from the inoculation to 2 or 4 weeks after infection and the second from 4 to 12 weeks. Yeast appear to be contained within the first week of infection and subsequently are also absent from macerated lung, differently from the model of BALB/c mice infected with ATCC Pb60855 conidia in which CFUs were detected up to week 16 post-infection. We found a mixed Th1/Th2 pattern (IL-12p70, IL-23, IL-10) in bronchoalveolar lavage and lung, with the predominance of proinflammatory cytokines in the first phase and predominance of regulatory Th2 cytokines in the second phase, reproducing findings of P. brasiliensis infection models produced with both conidia and yeast. However, in the first period of the disease in C57BL/6 mice there was a higher inflammatory burden, reflected by the high cytokine levels (TNF-alpha, MIP-2 and GM-CSF), the latter in particular because it was positively regulated both in vitro and vivo), that persisted through the chronic period. We also observed that starting from 48 hours postinfection to the chronic period there were increased levels of IL-12p70, which together with IL-23 appeared to be responsible for the reduction of infection in the late period. This is the first time that an experimental model with P. lutzii (Pb01) is described, showing an immunological profile with only slight differences compared to the P. brasiliensis model. The present study details important aspects of the pathogenesis of the disease due to different species of Paracoccidioides and helps to understand the different forms of presentation in experimental models

Our current system of research and production is no longer suitable for solving the problems we face today. As climate change threatens our cities and livelihoods, the global economic system preys on the weak. A more responsive, equitable, and resilient system needs to be implemented. Our post industrial cities are both products and victims of the boom-bust economies employed for the last few centuries. While some communities have survived by converting to retail and services based economies, others have not been so fortunate and have become run-down husks of their former bustling selves. The key to revitalizing these cities is to create new industries that empower people, unlike the service economies that deride and devalue them. Peer to Peer (P2P) development models like open source software communities create platforms for people to collaborate on projects and share resources. On the scale of cities, the goal is to stimulate the growth of closed loop, local, micro-economies that are inherently more stable than traditional, centralized economic models.Commons Based Peer Production (CBPP) is a term coined by Professor Yochai Benkler at Harvard Law School. It describes a new model of socio-economic production in which the labor of large numbers of people is coordinated (usually with the aid of the Internet) mostly without traditional hierarchical organization. It is based on low thresholds for participation, freely available modular tasks, and community verification of quality (peer governance). CBPP usually only applies to intellectual output, from software to libraries of quantitative data to human-readable documents (manuals, books, encyclopedias, reviews, blogs, periodicals, and more); however, this system can be adapted for physical manufacturing. A P2P system of development for material goods must be explored through the production of a common resource. Mycelium is the “roots” of fungi. It can be grown anywhere with agricultural refuse as a substrate. It has properties that make it ideal for building insulation and it is environmentally innocuous. It is Cradle to Cradle certified, and it requires little specialized equipment to produce. As a consumer product, it has had trouble gaining traction in a notoriously stubborn market dominated by hydrocarbon based market leaders like extruded polystyrene (XPS). Mycelium products are ripe for development as a regenerative building material. The goal is to increase the R-value of the material, decrease the cost of manufacturing, and carve out a market for this extraordinary product. The purpose of applying a CBPP approach is to increase the speed of development and aid in market penetration. The strategy is to decentralize manufacturing of and experimentation with the product. This requires a robust network of production nodes. Essentially, this involves setting up franchises in select markets (like the Pioneer Valley), where there is a strong interest in local, sustainable products. The nodes would be small cooperative businesses that are licensed to produce the material as well as collect data on the manufacturing and performance of mycelium insulation. The data will then be used to improve the production process. The bulk of the thesis is in designing one such node in Greenfield, MA, located adjacent to the new John W. Olver Transit Center on Bank Row St.