Programa de Pós-graduação em Ciências Biológicas

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Enumeração, isolamento e identificação de bactérias aeróbias psicrotróficas viáveis do estômago e do ceco de Tartaruga da Amazônia (Podocnemis expansa)
(Universidade Federal de Goiás, 1997-12-19) Oliveira, Wilian Pires de; Mesquita, Albenones José de; http://lattes.cnpq.br/3180029815183858
Foram utilizados no presente experimento 480 Tartarugas da Amazonia (Podocnemis expansa) mantidas em 16 caixas d’água, distribuídas em quatro grupos. Cada grupo, recebeu um tipo de ração contendo respectivamente 5,5, 7,0, 8,5 e 10,0% de fibra bruta. Cada caixa recebeu 30 animais recém-nascidos, oriundos da natureza e colhidos no rio Araguaia, na região compreendida entre o Porto de Luiz Alves e a Ponta Sul da Ilha do Bananal. De cada uma das 16 parcelas experimentais foram retirados aleatoriamente dois animais, para colheita de material do estômago e do ceco. Estas colheitas foram feitas aos 15, 89 e 144 dias de idade, totalizando 96 amostras. A análise microbiológica das amostras, permitiu isolar e identificar 17 gêneros de microrganismos aeróbios psicrotróficos viáveis nos conteúdos das visceras analisadas: Pseudomonas, Bacillus, Proteus, Enterobacter, Serratia, Micrococcus, Streptococcus, Flavobacerium, Citrobacter, Corynebacterium, Staphylococcus, Aeromonas, Hafnia, Acinetobacter, Salmonella, Edwardsiella e Klebsiella. O teste de coloração pelo método de Gram das bactérias, nas diferentes idades, revelou que o número de gêneros Gram-positivos permaneceu o mesmo, enquanto que o número de gêneros Gram-negativos reduziu com o aumento da idade. A temperatura da água foi tomada diariamente, e as médias obtidas variaram entre 24,5 e 26,1C. Quanto às características químicas da água representadas pelo pH, O2, CO2 e CaCO3 verificou-se que encontraram-se valores dentro dos limites adequados para organismos aquáticos. As diferentes rações fornecidas aos animais durante a fase experimental não influenciaram significativamente na microbiota, ao nível de 5% de probabilidade. Diferenças significativas, entretanto ocorreram na frequência de gêneros de microrganismos quando foram analisados as variáveis idades e órgãos.
Avaliação da atividade mutagênica e antimutagênica da Annona crassiflora Mart. (Araticum) pelo teste do micronúcleo em camundongos Mus musculus
(Universidade Federal de Goiás, 2001-08-23) FERREIRA, Francinez Linhares; CHEN, Lee Chen; http://lattes.cnpq.br/4621907105842007
RESUMO CAPÍTULO I The araticum (Annona crassiflora Mart.) is a typical brazilian plant found in cerrados of Brazil. This plant contains acetogenins that presents cytotoxic, antitumorigenic, antiparasitic and antimicrobial properties. Its leaves, barks, fruits and seeds are used by the population as therapeutic medicine to treat several diseases as diarrhoea, rheumatism and syphilis. In the present study we have evaluated the mutagenic activity of the crude ethanolic extract of leaves of araticum though quantification of micronuclei induced in mice bone marrow. Doses of 10 mg/kg (5% of LD50), 20 mg/kg (10% of LD50), 50 mg/kg (25% of LD50), 100 mg/kg (50% of LD50) and 160 mg/kg (80% of LD50) were applied i.p. in mice Mus musculus, in groups of 5 (five) animals for each dose. The cytological preparations were made according to Heddle`s methodology. For all the applied doses, frequency of micronucleated polychromatic erythrocytes (MNPCE) was evaluated after 24, 48 and 72 h of treatment. The cytotoxicity was evaluated by ratio the between numbers of polychromatic and normochromatic erythrocytes (PCE/NCE). Ours results indicated an absence of significantly increased of micronuclei for all the doses tested (p>0,01). Thus, we concluded that the phytoterapic araticum did not present mutagenic activity under our experimental conditions. However, the toxic activity was observed upon analysis of LD50 and PCE/NCE ratio. RESUMO CAPÍTULO II Annona crassiflora Mart. (araticum) is a typical brazilian plant found in cerrados of Brazil. This plant contains acetogenins that presents cytotoxic, antitumorigenic, antiparasitc and antimicrobial properties. Its leaves, barks, fruits and seeds are used by the population as therapeutic medicine to treat several diseases as diarrhoea, rheumatism and syphilis. In the present study we have evaluated the antimutagenic activity of the crude ethanolic extract of leaves of araticum through quantification of micronuclei induced in mice bone marrow. Doses of the extract (10mg/kg, 20mg/kg, 50mg/kg, 100mg/kg and 160 mg/kg) and MMC (4mg/kg) were co-applied i.p in mice Mus musculus in groups of 5 (five) animals for each dose. The cytological preparations were made in according to Heddle s methodology. For all the applied doses, frequency of micronucleated polychomatic erythrocytes (MNPCE) was evaluated after 36 h of treatment. The obtained results indicated that the crude ethanol extract from leaves ay doses of 20 mg/kg, 50 mg/kg and 100 mg/kg inhibited significantly reduced frequencies of micronuclei (P<0.01). Thus, we concluded that the phytoterapic araticum exerted an antimutagenic effect.
Avaliação dos efeitos ambientais da vegetação urbana sobre a qualidade de vida em Goiânia
(Universidade Federal de Goiás, 2001-08-31) Martins Júnior, Osmar Pires; Romão, Patrícia de Araújo; http://lattes.cnpq.br/0188184635964559; Brandão, Divino; http://lattes.cnpq.br/1614480825290154; Brandão, Divino; Casseti, Valter; Castro, Tomás de Aquino Portes e
This project is a study of the eco-system of the city of Goiânia (Goiás). Goiânia’s first Directive Plan was created according to the Garden City of Howard, one of the most important urban conceptions in the word. Using this plan and its urbanist evolution, one has tried to make a preliminary identification of the social agents, which produced the urban spaces of Goiânia. A classification and qualification of the open spaces and “green areas” of the city has been made. The quantity of vegetation (m².inhab.-1 ) and its quality (typing), when distributed adequately is important to the preservation of the urban eco-system, having important environmental effects on the quality of life of the population. The “green area” rate (Índice de Área Verde – IAV) is, therefore, one of the indicators of urban development. Changes in the open spaces, as indicators of IAV, in relation with changes in demographic density, indicative of human presence in the environment, have allowed the prevision of the amount of “green area” per urban inhabitant. The IAV calculated for Goiânia is 100,25 m². inhab.-1 , having suffered a per capita reduction of 17,68% in relation to the IAV established in the original city plan in 1938. In the next fifteen years it is possible forecast that the IAV will be 54,4% smaller than it is at the moment, decreasing to 45,71 m².inhab.-1 , if the privatization policy regarding publics spaces should persist. The “green areas” are unalienable and imprescriptible public property destined for common use. In spite of this, these areas have been decreasing over the years. The degree of deterioration of public assets has been calculated and constitutes a factor in the decay in urban life quality. The adoption of a programme to register and monitor the environmental assets of Goiânia is suggested, among other measures.
Caracterização Molecular Do Plasmídeo pLK39 Extraído De Uma Bactéria Endofítica Isolada De Solanum Lycocarpum
(Universidade Federal de Goiás, 2002-02-28) OLIVEIRA, Vera Lúcia Cardoso de; BATAUS, Luiz Artur Mendes; http://lattes.cnpq.br/5637230378599476
The characterization of the plasmid pLK39.was the aim of work.pLK39 was isolated from a endophytic Gram-negative, bactéria isolated from leaves form Solanum lycocarpum (Lobeira). In order to obtain a selection marker which would characterization in Escherichia coli the kanamycin, resistence gene from pUC4K. was subcloned into the PstIsite of pLK39. The characterization of PLK39 was basedon studies of stability, incompatibility, determination of the copy number and through DNA sequencing. The stability was carried out in Escherichia coli XL10 cells. Cells transformed with pLK39 were inoculated in Lúria broth containing Kanamycin (50μg/ml) during24 hours. After 24 hours of incubation one sample of the culture was in medim with and without selective pressure, and inoculated in Lúria broth without pressure médium, for 240 generations. After 240 generations 81,5% of cells maintained the plasmid, showing that pLK39is highly stable in Escherichia coli to make the incompatibility test, cells of Escherichia coli XL10 were co-transformed with pLK39/pUC18; pBR322 and pLK39/pACYC184. After 240 generations, the plasmid pLK39 iof detected in all systems used, showing the compatibility of pLK39 with the plasmids tested. The copý number pLK39 was estimated by the intensity of plasmidial bands in agarose gel, electrophoresis using a photodocumentation and analysis system (KODAK EDAS). The copý number of pLK39 was estimated as 25 copies per cell. PLK39 was digested with Sau3AI restriction and subcloned into the BamHI site of plasmid pUC18. The recombinant clones were sequenced and 1637 pb reading fragment was obtained. This sequence showed high homology with pSW200, pSW100, pEC3, pUCD5000 and pBERT, which were isolated from Gram-negative bacteria. This sequence possesses two possible genes. The ORF I showed high homology with mobB gene from plasmid pSW200 (96%), pSW100 (96%), pEC3 (94%), pUCD5000 (94%) and pBERT (87%). The ORF II showed homology with mobD gene from plasmids pSW200 (92%), pSW100 (90%), pEC3 (90%) and PUCD5000 (89%).
Caracterização do gene ftsH de Streptomyces sp Y7
(Universidade Federal de Goiás, 2002-12-14) Paixão, Cinthia Ferreira da; Bataus, Luiz Artur Mendes; http://lattes.cnpq.br/5637230378599476
The actinomycets are Gram-positive bacterias, aerobic with rich DNA in G+C (larger than 60%) and immobile. They are found practically in all the environment, forming ramified filaments or hyphae that persist in the mycelium form. The Streptomyces constitutes 90% of the isolated actinomycets of the soils, in spite of they are also found in aquatic atmospheres and interior of some plants. They stand out for the diversity of production of hidrolytics enzymes and antibiotics, 70% of the know antibiotics are produced by those microorganisms. Aiming to clone genes with biotechnological interest, a genomic libraries of the Streptomyces sp Y7 isolated of the soil of Cerrado was constructed. After the analyses of the sequences of the genomics libraries of Streptomyces sp Y7, it was selected a plasmid named pFS8, that displayed similarity with ftsH genes. The ftsH gene encodes a metalloprotease ATPase and Zn+2 dependent, belongs to the AAA family (ATPases associated with a variety of cellular activities). It is involved with several cellular functions such as secretory proteins export and degradation of transcriptional factors (sigma 32 and Lambda CII). The data of sequencing showed that the ftsH gene was incomplete. In order to characterize if the product of this gene showed biological activity, it was made tests to evaluate the functionality of the fusions proteins in an ftsH-negative E.coli strain AR3291. AR3291 cells transformed with this plasmid showed a general growth advantage upon the cells AR3291. The protein produced did not present toxicant effects for cells AR3289, which had normal ftsH gene. The truncated protein obtained was also analyzed to prediction of the structure “coiled-coil”, that is common to the other FtsH studied, and the results showed those truncated proteins did nor form “coiled-coil” structure. We also tested whether fusion proteins decreased or inhibited the defective transfer of citosolic proteins.
Caracterização limnológica e distribuição espacial de insetos aquáticos, das ordens Ephemeroptera, Plecoptera e Trichoptera, na bacia do Rio Meia Ponte, Goiás, Brasil
(Universidade Federal de Goiás, 2003-11-05) SILVEIRA, Flávio Luiz de Souza; OLIVEIRA, Leandro Gonçalves; http://lattes.cnpq.br/8837912214958454
The Meia Ponte River is one of the most important water sources in Goiás State, supplying water for almost half of its population. However, its watershed has been strongly affected by intense agriculture practice, cattle breeding, and industrial and domestic effluent discharges. In this paper, 34 sites in this watershed were chosen to the development of a limnological study. In the first chapter, the limnological characterization of these sites is presented, including 20 physical-chemical, microbiological and physiographic variables, during the rainy and dry seasons of 2001. The sites located at the Meia Ponte River downstream from Goiânia municipality showed the worst water quality. Temporal and spatial patterns were observed. In the second chapter, the influence of abiotic factors on Ephemeroptera, Plecoptera and Trichoptera community was accessed during the dry season of 2001. Physiographic variables were the most important factor for community variation (mainly substrate type and water velocity). Physical-chemical variables had a secondary effect on community structure.
Cultivo de embriões bovinos em estágio de blastocisto em meio sequencial, após maturação e fertilização de oócitos in vitro, de novilhas e vacas velhas: comparação entre métodos de congelamento lento, computadorizado e ultra-rápido
(Universidade Federal de Goiás, 2004-03-01) Rocha, Jane Porfírio; Freneau, Gustavo Eduardo; http://lattes.cnpq.br/9867769335357011
Studied in this work maturation in vitro oocytes collected from ovaries obtained from a slaughterhouse, without any hormonal stimulation and, comparing these data at different stages of maturation between heifers and old cows. The culture medium used for maturation was the In Vitro Fertilization (Scandinavian IVF Science AB. Sweden), where similar results were observed for the two age, presenting a significant correlation (r = 0.9991 and p <0.0001) for stages of maturation, despite the maturing rate for new cows have better 50.5%, and for the old cows of 31.3%, there was no statistical difference. The same is repeated for the fertilization of young cows to 98.1% and 86% for old cows. For the blastocyst formation rate, 17.14% to 22.58% and old cows to young cows. We tested two types of cryopreservation for embryos in the blastocyst stage, the ultra-fast freezing, using dimethyl sulfoxide-cryoprotector (DMSO), and the computerized slow freezing with cryo chambers - Australian, using glycerol as cryoprotector. Embryos at the new cows DMSO showed a 14% survival rate and 73% glycerol, and the old cows were 31% DMSO and 63% glycerol. The rehydration rate of re-expansion of blastocysts after thawing was 15% with 15% DMSO and 38% glycerol for old cows and 14% DMSO and 55% glycerol for new cows. Spontaneous Hatching was observed only for the embryos thawed freezing with glycerol for young cows. In the statistical calculation was no significance between the results of survival, to total frozen and thawed embryos, between the slow freezing methods and the ultra-fast slow cryopreservation method using glycerol as a cryoprotector.
AVALIAÇÃO DA ATIVIDADE ANTIINFLAMATÓRIA DA FRAÇÃO ALCALOÍDICA DO FRUTO DE Solanum lycocarpum A.St.-Hil. (LOBEIRA)
(Universidade Federal de Goiás, 2004-09-10) VIEIRA JÚNIOR, Geraldo; PAULA, José Realino de; http://lattes.cnpq.br/3191837532986128
Solanum lycocarpum A. St.-Hil., popularly known as lobeira (wolf-fruit), is easily found in the Brazilian savanna. It is characterized as a bushy plant with average height of about 5 meters (16 feet), fragile branches, featuring a nearly round berry type of fruit, with diameter that ranges from 8 to 15 cm (3.1 to 5.9 inches), yellowish green color even when ripe, fleshy and juicy pulp. The lobeira is used in folk medicine for several therapeutic purposes such as bronchitis, worm diseases, diabetes and ulcer. A study with ethanolic extract of fruit (EEF) of the lobeira showed analgesic and antidermatogenic activity in the experimental samples of acetic acid-induced abdominal writhing and ear edema. In the present work were performed the morphoanatomic study of the fruit and the antiinflammatory evaluation of the alkaloidic fraction (FA) acquired through the EEF. The morphoanatomic analysis of the fruit pointed out the existence of elements that allow its characterization, like the stellate trichomes, present in leaves of the same species as well. The FA was obtained by acid extraction of the EEF, being defined by thin-layer chromatography the presence of the glycoalkaloid solamargine, found in other species of the Solanum genus. The antiinflamatory activity was estimated in vivo through the ear edema method induced by the croton oil and by the leukocyte migration in the peritonitis induced by carrageenin. It was estimated in vitro the FA ability to restrain the phospholipase A2 action in snake poison. The prior treatment of animals with FA in doses of 50 to 100mg/kg (s.c.) reduced the ear edema formation, and in doses of 30 a 300mg/kg (s.c. and p.o.) decreased the leukocyte migration. The experimental models in vivo demonstrated that the FA displayed characteristic results of an antiinflamatory activity similar to the one shown by the glycocorticoids. The results obtained with the FA could be outcome of the glycoalkaloid presence and also indicate that the FA has the active principles responsible for the biological activity verified in EEF. The FA has not presented inhibitive activity of phospholipase A2 that could be detected by in vitro method that was applied. It becomes necessary the purification of the glycoalkaloid and the completion of specific trials aiming to confirm the antiinflamatory action as well as the participating mechanisms.
Caracterização Molecular e Expressão Heteróloga de um cDNA Codificante para Tiorredoxina do fungo patogênico humano Paracoccidioides brasiliensis
(Universidade Federal de Goiás, 2006-08-31) DOMINGOS, Fernanda de Castro; JESUÍNO, Rosália Santos Amorim; http://lattes.cnpq.br/5113656623817587
The temperature-dependent dimorphic fungus Paracoccidioides brasiliensis is the etiological agent of Paracoccidioidomycosis (PCM), a human systemic mycosis highly prevalent in countries of Latin America. P. brasiliensis is subjected to different insults from human host, such as oxidative stress caused by reactive oxygen species produced by the host during the infection. Thioredoxin (TRX) is an intracellular redox protein that is required to maintain redox homeostasis in response to both reductive and oxidative stress conditions in several organisms. We report here the characterization of a 811 bp cDNA Pbtrx1, encoding a PbTRX1 of 116 amino acids, with a predicted molecular mass of 12 kDa and pI 5.2. This putative protein presented one highly conserved active site motif (WCGPC) between TRXs from several organisms. The phylogenetic analysis performed with PbTRX1 and TRXs from other organisms, putted P. brasiliensis in the fungi clade. We also performed the prediction of the secondary structure of PbTRX1 that shows a pattern characteristic of the open twisted alpha/beta, similar to TRX secondary structures described in other fungus. In order to obtain the recombinant PbTRX1, the expression construct pGEX-4T-3-trx1 was introduced into Escherichia coli cells and the expression and purification of the recombinant protein was obtained. The recPbTRX1 and PbTRX1 from yeast cells extract were found to catalyze the reduction of insulin. However the PbTRX1 from yeast cells extract treated with H2O2 showed highly insulin reduction activity than the yeast cells no treated. PbTRX1 was detected by Western blotting in the extracts from yeast cells growth and from mycelium to yeast transition. The yeast cells growth was significantly inhibited by H2O2; however the mycelium to yeast transition was little affected by this oxidant. Semi-quantitative RT-PCR was employed to analysis the expression of Pbtrx1 gene in response to H2O2. The level of Pbtrx1 transcripts was higher in yeast cells treated with H2O2 than in yeast cells no treated. To realize how P. brasiliensis deals with oxidative stress is essential to understand the mechanisms involved in its survival in the host. It may be possible that PbTRX1 enhances survival of P. brasiliensis in the host, protecting the fungus against the reactive oxygen species and allowing, in this way, the progress of the infection.
Análise epidemiológica, molecular e citogenética do câncer de hipofaringe em Goiânia
(Universidade Federal de Goiás, 2006-09-29) Costa, Cesar Augusto Sam Tiago Vilanova; Cruz, Aparecido Divino da; http://lattes.cnpq.br/7868817504129985
Hypo pharynx is the under region of pharynx and serves both respiratory and digestive tract, acting as a passageway to air and food. It’s composed by squamous nonkeratinized epithelial tissue. Statistical studies indicates that 85 to 90% of neoplasies rush pyriform sinus and are characterized by fast dissemination and metastization. Epidemiological trends of hypo pharynx cancer in Goiania on period 1998 to 2003 analyzing data derived from Cancer Registry of Association for Cancer Combat of Goias, shows that the relative survival rate for men diagnosed with hypo pharynx cancer was 38% past 60 months while women presented a survival rate of 100%. Surgery was the treatments that present fewer rates (34%) and chemotherapy was associated to the most survival rate (54%). Patients that use tobacco presented a survival rate of 43% while non-tabagists present a rate of 60% of surviving. Relative survival rate for patients diagnosed with hypo pharynx tumor was 38% and estimated censure for 5 years was just 5%. Survival rate was extremely decreased due to 45% of cases were diagnosed in advanced stages of disease. It is important to point out that preventive campaign is important to evade the increasing of these pathologies that are related to behavior of tabagism and etilism. Present study also evaluates cytogenetically individuals diagnosed with hypo pharynx cancer. Cytogenetic analysis of exfoliated cells is largely used to evaluate cytotoxic and genotoxic alterations on people exposed to potentially mutagens. A number of 36 individuals were analyzed, representing case group, and 36 healthy individuals composed control group. Data was collected and analysis was performed to estimate main nuclear alterations. Standard protocols were adopted to collect and counting cells. Micronuclei test was used to determinate the frequency of cytotoxic and genotoxic alterations. Besides than micronuclei others alterations like broken-eggs, binucleated cells and piknotic nuclei are good markers of cell alterations because they are resulted from cytotoxic events frequently related to occupational exposure. Behaviors like tabagism, etilism and exposition time were also investigated to correlate cells alterations. The correlation between micronuclei frequencies of case and control group test doesn’t show statistic difference. Broken egg and binucleated cells were the most observed anomalies on case group and showed up significant statistic difference (p=0.03). Results indicates a high grade of cytotoxic anomalies on individuals with hypo pharynx cancer, specially in tabagists and chronic etilists individuals, behaviors that are correlated to induct and promote tumors in susceptible people due their capabilities to affect cell cycle. Studies about relationship between Human Papillomavirus (HPV) and cancer development are performed since 80’s. Although the correlation of HPV and cervical cancer are well established, the possible role of HPV and head and neck cancer are still unclear. Among controversial factors of HPV and carcinogenesis of squamous epithelial cells, there is a viral incidence that oscillates from 0 to almost 100% on studies that includes a myriad of viral genome detect methods, such Blotting and PCR. Importance of HPV infection and epithelial carcinogenesis is related to capabilities of High risk HPV 16 and 18 to promote cell alterations, by tumor suppressor proteins inactivation, tumor suppressor genes blockage and insertions of HPV oncogenes. On present study, HPV detection and typing was performed on tumors derived from hypo pharynx of 24 patients diagnosed with cancer (CID-O: C12,0 to C13,9). All samples was submitted to PCR with generic primers MY09/11 and GP05/06, as specific primers to HPV 6, 11, 16, 18, 33, 35, 45 and 58. Results shows that on 9 of 24 samples, HPV were detected by generic primers (a rate of 37,5%) and HPV types 16, 18 and 45 were identified, with a sample presenting coinfection of HPV 16 and 18.
Fluorescência da clorofila a e liberação de O2 em diferentes variedades de feijão (Phaseolus vulgaris) e soja (Glycine Max(L.)Merr.) após aplicação de sulfato de amônio
(Universidade Federal de Goiás, 2006-11-25) Silva, Kelly Pereira da; Castro, Tomás de Aquino Portes e; http://lattes.cnpq.br/7313401367533383; Castro, Tomás de Aquino Portes e; Fageria, Nand Kumar; Didonet, Agostinho Dirceu
The productivity of vegetables is influenced by weather, physiology and genetics associated by photosynthesis, by the division of photoassimilated, and by the fixation of N. The analysis of fluorescence of clorophyll allows the study of the characteristics related with the capability of absorption and transference of the luminous energy in the chain of electron transport. The aim of the present research was to test three varieties of kidney bean and three varieties of soy bean influenced by nitrogen in the exchanges of O2 and the fotochemical efficiency (Ef), using techniques of emission of fluorescence and O2 envolved during photosynthesis, comparing each other. The determinated variants were: minimum fluorescence (F0), maximum fluorescence (Fm), terminal fluorescence (Ft), variable fluorescence (Fv), efficiency quencher (Eq) and fotochemical efficiency (Ef). The O2 envolved during photosynthesis was avaliated. The plants were submitted to the recommended level of N and the determinations of fluorescence emission (F0, Fm, Ft, Fv, Fv/Fm) and the O2 envolved during photosynthesis were realized one day after the application of NH4+ like ammonium sulfate. Related to fluorescence there was not difference between the varieties of kidney bean and soy bean. When it was analised the application of NH4+ it was observed that there was difference in the statistic of soy bean. It was observed the O2 envolved during photosynthesis in the variety of kidney bean Pérola ( 1,50 mol de O2 m –2 s –1) was greater than the other and in the variety of soy bean Monsoy ( 1,58 mol de O2 m –2 s –1) was greater than the other. Additional index terms: O2 evolution, Phaseolus vulgaris, Glycine Max, fluorescence, fotochemical efficiency, quencher efficiency.
Anotação e caracterização preliminar de genes de celulose sintase em diferentes espécies de Eucalyptus
(Universidade Federal de Goiás, 2006-12-01) SALAZAR, Marcela Mendes; COELHO, Alexandre Siqueira Guedes; http://lattes.cnpq.br/0840926305216925
Cellulose is the world s most abundant polymer, being the main constituent of plant biomass. Genes from CesA family, that encodes the cellulose synthase enzyme, was identified in several plant species, especially in Arabidopsis thaliana, in which three of them (AtCesA4, AtCesA7 e AtCesA8) were associated with secondary cell wall cellulose production. Eucalyptus species represent an important target of genetic improvement studies, since it is the most planted forest genus in the world, beyond deserving a special place in the international market of cellulose and paper. The molecular breeding technology enables that gene characterization can be applied to forest species genetic improvement programs with the aim to improve the quality and productivity of its products. In this work, 320 Eucalyptus ESTs, separated in four genes related with cellulose production in secondary cell wall, using AtCesA4, AtCesA7 e AtCesA8 genes as reference. For these genes, primers were designed in order to screen an Eucalyptus BACs library. An emphasis was given to genes that are orthologs to AtCesA7 from Arabidopsis thaliana for witch it was sequenced a 1197 base pairs region from the BACs library and this served as a support to expression level studies of this genes in different species/tissues, showing that this is preferentially expressed in xylem and weakly expressed in leaves. The expression level of this gene was higher in E. urophylla than in other species studied. Sequences from approximately 500bp was obtained from different Eucalyptus species and in these, with one intron between two exons, the amount of SNPs in the intron (4), as waited, was higher than that found in exons (1 in each), although the intron nucleotide diversity index (π) (0,0824) were less than in the exon (0,2029). In this manner, one expects that this work can contribute for one better understanding of the mechanisms involved in biosynthesis and regulation of the cellulose pathway in Eucalyptus species, as well as subsidize genetic mapping studies and linkage disequilibrium analysis for this genus.
Mapeamento de QTLs para teor de proteína em feijoeirocomum (Phaseolus vulgaris L.)
(Universidade Federal de Goiás, 2006-12-15) LEÃO, Ariane Castro Mendes; CARNEIRO, Monalisa Sampaio; http://lattes.cnpq.br/2696490871291334
The common bean besides being one of the basic meals of brazilian´s population, it is one of the main products that provide protein in the nutritional diet from the society share which is economically less favorable. The identification of molecular markers linked to controlling genes of the protein content in common beans (Phaseolus vulgaris L.) is a very important tool to help breeding programs, raising the efficiency and agility. This way, this work was made with two main goals: a) to map SSR and RAPD markers linked to loci (QTLs) that control protein content in two generations of a segregating population of common beans and b) to compare detection procedures of markers linked to QTLs using the ANOVA method and the process of interval mapping. For that reason, 94 families were taken from the F2 generation and 90 families from the F2:3 generation derived from the cross of genitors CNFC 7812 e CNFC 8056. Results indicated that there is the possibility of identifying molecular markers related to protein content in common beans, utilizing both detection procedures. The ANOVA method identified a greater number of QTLslinked markers than the process of interval mapping in both generations. There was coincidence between the identified loci obtained with the two methods for each generation. Loci that were associated with protein content were different for the F2 and F2:3 generations. However, there was a stable detection of a genomic region of the linkage group 4, indicating a possible role of this region of the common bean genome in the control of seed protein content. The proportion of the trait s phenotypic variation explained by QTLs varied from 5,5% to 9,5%, considering both generation.
Morfologia do fígado e das brânquias do guaru (Poecilia vivipara) expostos às concentrações agudas do herbicida Roundup original (glifosato(N-(fosfonometil) glicina))
(Universidade Federal de Goiás, 2007-08-20) Leão, Michelle Furquim; Sabóia-Morais, Simone Maria Teixeira de; http://lattes.cnpq.br/6723881044959716
The sharp toxicity of the herbicide original Roundup (Glifosato), one of the more acquaintances and maybe one of the more used dessecantes in direct planting in the farmings in the area Centro Oeste and in every country now, it was investigated through the effects detected in the fish Poecilia viviparous...
Morfo-anatomia e fitoquímica de Cymbopogon densiflorus (Steud.) Stapf e Cymbopogon nardus (L.) Rendle (Poaceae: Panicoideae)
(Universidade Federal de Goiás, 2007-09-28) BARBOSA, Lília Cristina de Souza; PAULA, José Realino de; http://lattes.cnpq.br/3191837532986128; REZENDE, Maria Helena; http://lattes.cnpq.br/5093753722360659
The genus Cymbopogon Sprengel belong to the Poaceae family and it has 40 species distributed in Tropical and Subtropical Africa, Asia and Australia, although some species went introduced in America. Many species of this genus are cultivated for the extraction of essential oil, from their leaves, with large medicinal, food and industrial importance. The species in focus, Cymbopogon densiflorus (Steud.) Stapf and C. nardus (L.) Rendle are originated from Africa and Asia, respectively. This research had as objective, to broaden the knowledge about the species C. densiflorus and C. nardus, by the morphological and anatomy studies from leaves and culms, phytochemical analysis and essential oil analysis from the leaves. Anatomical studies have been of relevant importance to the pharmacognosy researches, mainly for the identification of many vegetal raw materials. Several times, these raw materials are known by the same popular name or then, they are commercialized with contaminated agents or with other parts of the specie. Through of anatomical analysis, it was checked commons characters, such as leaf lamina and sheath amphistomata, stomatas with guard cells dumbbell and subsidiary cells dome-shape, rares in adaxial surface and abundant in abaxial surface, predominated in intercostal zones. The adaxial and abaxial surfaces had long cells and short cells: cork and dumb-bell and cross-shaped silica cells, these last it is placed in costal zones; macro-hairs and micro-hairs abundant in abaxial surface. In the leaf lamina, bulliforms cells are presents in adaxial surface, they were alternated with fibers in the costal zones and the mesophyll is homogeneous with chlorenchyma radiated to the bundle sheaths and arm cells with walls invaginated that they determined the intervenal distance by one or three cells, characterized Kranz anatomy. Bundle sheaths collateral, of 1st, 2nd and 3rd orders with single vascular bundle sheaths. The cap region is constituted by sclerenchyma and the epidermis has silica cells. However, both species had different anatomical features, as the form of midrib, in the leaf laminas; C. densiflorus showed colourless parenchyma cells in the mesophyll of leaf sheaths, that they do not exist in C. nardus. In the culms, numbers of metaxylem vessels in the each side of protoxylem vessels in vascular bundles: 1, in C. densiflorus, 2 and 3, in C. nardus; and the presence of sclerenchyma cylinder and fistula in C. nardus, absent characters in C. densiflorus. Moreover, in C. densiflorus, while C. nardus showed these characters. The preliminary phytochemistry analysis C. densiflorus and C. nardus leaves evidencied flavonoids, saponins, coumarins and traces of cardioactive glycosides. In the essential oil analysis, C. densiflorus leaves showed trans-p-mentha-1(7),8-dien-2-ol, trans-p-mentha-2-8-dien-1-ol, cis-carveol and cis-p-mentha-2-8-dien-1-ol as majority constituents; while C. nardus leaves had geraniol, citronellol and citronellal. The anatomical characters observed can be important to the taxonomic determinations of species studied, in the genus. Through the results found, it verifies the phytotherapics potential of both species. Future researches in isolation and purify of the secondary metabolites, pharmacologics and toxicologics analysis of extracts and of the essential oil, it will be important to assure the therapeutic efficiency of these.
Caracterização de um Antígeno Rico em Prolina(PRA/Ag2) do fungo patogênico humano Paracoccidioides brasiliensis
(Universidade Federal de Goiás, 2008-01-31) CASTRO, Kelly Pacheco de; SOARES, Célia Maria de Almeida; http://lattes.cnpq.br/8539946335852637
The dimorphic fungus Paracoccidioides brasiliensis is the causative agent of the most frequent systemic mycosis in Latin America. In humans, infection starts by inhalation of fungal propagules, which reach the pulmonary epithelium and differentiate into the yeast parasitic phase. Here we describe the characterization of a proline-rich protein (PRA/Ag2) homologue of P. brasiliensis, a predictable cell wall protein, first identified in Coccidioides immitis. The protein, the cDNA and genomic sequences were analyzed. Southern blot analysis suggested that there is one copy of the gene in P. brasiliensis. The cloned cDNA was expressed in Escherichia coli and the purified rPbPRA/Ag2 was used to obtain polyclonal antibody. The purified recombinant protein was recognized by sera of patients with proven paracoccidioidomycosis and not by sera of healthy individuals. Immunoelectron microscopy and biochemical studies demonstrated the presence of PbPRA/Ag2 in the fungal cell wall, linked through a GPIanchor. The expression of the Pbpra/ag2 gene was analyzed by real time PCR and results demonstrated developmental regulation in phases of P. brasiliensis, with a higher expression in the mycelium saprobic phase. The protein expression analyses corroborate the transcript levels.
ESTUDO DO POTENCIAL MUTAGÊNICO E ANTIMUTAGÊNICO DA Solanum paniculatum L. PELO TESTE DO MICRONÚCLEO EM CAMUNDONGOS
(Universidade Federal de Goiás, 2008-02-23) VIEIRA, Pabline Marinho; CHEN, Lee Chen; http://lattes.cnpq.br/4621907105842007
Solanum paniculatum L., popularly known as jurubeba, is a widespread plant species used in Brazilian folk medicine as a tonic, antifever agent, colagogue, bitter, and eupeptic to treat liver and gastric dysfunctions. The chemical composition of Solanum paniculatum has long been studied and many substances have been isolated from the entire plant, including alkaloids, resins, glucids and saponins. The aim of this study was to evaluate the mutagenic, antimutagenic and cytotoxic effects of jurubeba s ethanolic leaf and fruits extracts using micronucleus test in mice bone marrow. The experimental procedure was performed according Schimid (1975). Swiss mice were orally treated with three different concentrations (100, 200 or 300 mg/kg) of leaf or fruit extracts of S. paniculatum. To evaluate the antimutagenic activity were used the same doses of the extract simultaneously with a single dose of Mitomicin C (4mg/kg) i.p. Cytotoxicity was evaluated by the polychromatic and normochromatic erythrocytes ratio. Our results have indicated that the ethanolic leaf extract of S. paniculatum did not exhibit mutagenic effect in mice, but it showed antimutagenic effects in this extract towards MMC. The cytotoxicity was strongly demonstrated specially for higher doses. Meanwhile, the ethanolic fruit extract of S. paniculatum was mutagenic only in dose of 200 mg/Kg at time of 48 hours. It was showed the cytotoxic activity for higher doses. However, no antimutagenic effect was evidenced by fruit extract.
AVALIAÇÃO DAS ATIVIDADES GENOTÓXICA E ANTIGENOTÓXICA DE DUGUETIA FURFURACEA EM BACTÉRIAS E CAMUNDONGOS
(Universidade Federal de Goiás, 2008-02-25) SILVA, Carolina Ribeiro e; CHEN, Lee Chen; http://lattes.cnpq.br/4621907105842007
The plant Duguetia furfuracea (St. Hil.) Benth and Hook. f. (1862), belongs to the family Annonaceae, is popularly known as sofre‐do‐rim‐quem‐quer, araticum‐do‐cerrado and ata‐do‐mato. This plant occurres in several Brazilian states, as Amazonas, Bahia, Distrito Federal, Goiás, Minas Gerais, Mato Grosso, Mato Grosso do Sul and São Paulo. It is commonly used by people as anti‐rheumatic and to cure renal colic. Previous studies have described therapeutic activity of this plant with trypanocidal, antiplasmodial and antiprotozoal effects. Due to the large utilization of this plant by the local people, the present work aimed to evaluate the genotoxic/mutagenic, antigenotoxic/antimutagenic and cytotoxic activities of lyophilized leaf extract of D. furfuracea by the lisogenic induction test (Inductest) and the mice bone marrow micronucleus assay (Micronucleus Test). The lisogenic induction test was performed according Moreau et al., (1976) using E. coli WP2s(λ) and RJF013 strains. To evaluate the genotoxic and toxic activities, the doses employed were 1 mg, 2 mg, 5 mg e 10 mg, while to evaluate the antigenotoxic action the doses were of 0,5 mg, 1 mg, 2 mg, 5 mg e 10 mg co‐treated with 0,5 μg de mitomycin C. Micronucleus test in the bone marrow of mice was performed according to Schmid (1975). To evaluate mutagenic activity, mice were treated with three different doses of plant extract (100, 200 and 300 mg/Kg body weight), to evaluate antimutagenic activity, mice were treated with the same doses co‐administered with 4 mg/kg of mitomycin C. The genotoxic action was evaluated by the frequency of micronucleated polychromatic erythrocytes (MNPCE) and cytotoxicity was evaluated by the polychromatic and normochromatic erythrocytes ratio (PCE/NCE). The results obtained from the evaluation of mutagenicity of this plant showed that the extract of D. Furfuracea didn t increase significantly (p>0.05) the frequency of MNPCE compared to negative control. Cytotoxicity was evident in all applied doses (p<0.05). Concerning antimutagenicity, all doses of extract reduced significantly the frequency of MNPCE compared to the positive control group (p<0.05). In both tests, the results obtained demonstrated that the extract of D. furfuracea presented cytotoxicity; however, it didn t show genotoxic or mutagenic effects in bacteria and mice bone marrow respectively. In the other hand, it was observed a antigenotoxic and antimutagenic action.
Purificação parcial das quitinases, Pbcts1 e Pbcts2, do fungo Paracoccidiodes brasiliensis
(Universidade Federal de Goiás, 2008-04-03) SANTANA, Lidiane Aparecida da Penha; PEREIRA, Maristela; http://lattes.cnpq.br/1345781867765758
Paracoccidoides brasiliensis is a human pathogenic dimorphic fungus. The recombinant chitinase from P. brasiliensis, Pbcts1r, was overexpressed in Escherichia coli using pET-32a (+) as vector. The enzyme was produced as inclusion bodies and became soluble by Sarkosyl being purified by a single step using a Ni-NTA resin. Pbcts1r showed activity against 4-MU-(GlcNAc)3 and 4-MU-(GlcNAc)2, presenting a endochitinase activity. Immunoblot reaction with anti-Pbcts1r identified two proteins in yeast crude extract. A partial purification of P. brasiliensis yeast crude extract by cationic-exchange chromatography on HPLC revealed two different chitinases, Pbcts1 and Pbcts2, with molecular mass of 45 kDa and 34 kDa, respectively. Pbcts2 has exochitinase activity and Pbcts1 has endochitinase activities. Reactions with anti- Pbcts1r showed the presence of Pbcts1 and Pbcts2 in crude extracts of yeast and transition from mycelium to yeast. On mycelium crude extracts was found only Pbcts1 and on yeast cell wall extract only Pbcts2. Both proteins were found to be secreted by yeast parasitic phase showing their probable importance in the permanence of the fungus in the human host. Phylogenetic relationships between the orthologs Pbcts1 and the putative Pbcts2 indicated the presence of a common ancestral. During evolution, P. brasiliensis could have acquired Pbcts2 and Pbcts1 playing distinct roles in order to growth and survive in diverse environment on saprophytic and parasitic phases
Identificação e descrição morfoanatômica e farmacognóstica das folhas de Solamum Scuticum M. Nee e bioatividade de extrato bruto em microorganismos e da fração alcaloídica em células cultivadas da linhagem vero
(Universidade Federal de Goiás, 2008-04-16) MORAES, Leslivan Ubiratan de; PAULA, José Realino de; http://lattes.cnpq.br/3191837532986128; SABÓIA-MORAIS, Simone Maria Teixeira de; http://lattes.cnpq.br/6723881044959716
The jurubeba was identified that such species was Solanum scuticum M. Nee, from which pharmacognostic and morphanatomic data were not available to Goiás. Because of that, we tried some complementary data. For the morphoanatomic evaluation of the leaves some cuts were carried out as described by Kraus and Arduin (1997). For the phytochemical trial, it was used some methodologies as described by Costa (2001). With the phytochemical screening, were identified alkaloids, flavonoids, coumarins, anthraquinones and saponins in prospecting phytochemical. Both in phytochemical trial, as in the phytochemical prospection, it was found alkaloids. Due to the biological activities of these secondary metabolites, we obtained the alkaloid fraction, using the dust of the leaves, and we got the ethanol extract. The fractions were divided by polarity and they were evaluated by thin-layer chromatography. Microbiological evaluation was carried out to verify possible contaminants. Such evaluation did not reveal the presence of microorganisms, and it was raised the possibility of antimicrobiotic activity of the raw extract in twenty three strains of bacteria and in two yeasts. According to the antimicrobial tests, the extract presented some features, as the difficult of solubility in aqueous medium, and the best dilution was achieved in dimethyl sulfoxide (DMSO) 50%. These tests also demonstrated the difficult of the solubilization of the fractions, which it will be used in the experiments of Vero cells cultures. It was verified the low antimicrobial activity of the raw hidroethanolic extract. Of all the fractions, the alkaloid fraction presented the best solubility in DMSO 0.3%. So, it was proceeded the dilution and the evaluation of the alkaloid fraction in Vero cells, and it was evaluated morphology, viability and cellular proliferation. We finally verified the citotoxicity of the alkaloid fraction in the statistical analyses, and in the vitality test carried out by Trypan blue, and also in the morphological alterations compatible to citotoxity alterations.

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