Doutorado em Biotecnologia e Biodiversidade Rede Pró-Centro-Oeste (PRPG/UnB)

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    Detecção e análise de mutações de novo em pacientes com exposição parental à radiação ionizante de Césio-137 a partir de dados de genotipagem de polimorfismos de base única de alta densidade
    (Universidade Federal de Goiás, 2020-10-28) Leite Filho, Hugo Pereira; Caetano, Alexandre Rodrigues;; Silva, Cláudio Carlos da;; Silva, Cláudio Carlos da; Telles, Mariana Pires de Campos; Silva, Daniela de Melo e; Cruz, Alex Silva da; Gigonzac, Marc Alexandre Duarte
    In 1987, in the city of Goiânia, a series of unexpected events resulted in a serious radiological accident, generated by Césio-137. The mutagenic effects of ionizing radiation (IR) can lead to accumulation of mutations in children of irradiated parents. It was established that chromosomal microarray analysis (CMA), a cytogenomic technique for the detection of SNP in a wide spectrum of regions of the human genome. The use of cytogenetic assays based on high-density microarray of DNA allows identifying variations in SNPs and, consequently, genotyping them. In this study, using the GeneChip® HD® assay it was possible to establish the genotypes of SNPs in a population born from progenitors exposed to cesium-137 ionizing radiation. Mendelian germline deviations were used to estimate the rate of mutations induced by parental exposure in their offspring. The exposed group consisted of 11 families, of which at least one parent was directly exposed to Cesium-137 ionizing radiation, including a total of 37 individuals (11 couples and 15 children born after the accident). The absorbed dose for exposed individuals ranged from 0.2 to 0.5 Gray. A group of individuals not exposed to ionizing radiation was used as a control. This group consisted of 15 families from Goiás with no history of exposure to IR. The statistical tests used were: Shapiro-Wilk test, F test, regression analysis, clustering, and the main component analysis. All analyses were performed using the statistical package R, with a significance level of 5% (p<0.05). FMDM frequencies were estimated by case and control, representing 1.3x10-3, 0.9x10-3, respectively. Thus, the frequencies of FMDM showed statistically significant differences between the exposed and control groups (p<2x10-3, α = 0.5, Student-t). The F test to compare the variations of two samples (Case and Control) from populations with normal distribution showed that the variations (F=4.47; α = 0.5, p<8x10-3) between cases and controls were significantly different. In addition, the progeny of a population accidentally exposed to low doses of IR showed ~ 1.44x more de novo Mendelian deviations (MD) than healthy controls. In conclusion, the frequency of germ line/generation MD mutation may be useful to study human populations exposed to IR retrospectively, using the Mendelian deviation findings technique, it was possible to identify the origin of the parents, as well as the type of substitution and inform which variant suffered the mutation. Therefore, DM are potentially useful markers to discriminate parental exposure to IR.
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    Photon flux density and wavelengths alter the morphophysiological and chemical caracteristics of Anacardium othonianum Rizz. in vitro
    (Universidade Federal de Goiás, 2020-01-22) Rosa, Márcio; Pereira, Paulo Sérgio;; Rubio Neto, Aurélio;; Rubio Neto, Aurélio; Silva, Fabiano Guimarães; Pinto, José Eduardo Brasil Pereira; Miranda, Ricardo Motta; Batista, Priscila Ferreira
    Anacardium othonianum Rizzini is a native Cerrado species with great biotechnological potential. The selection of intensity and spectral quality may represent an alternative to optimize the physiological and productive characteristics of this species in vitro culture favoring its propagation. In this context, the objective was to define the best light conditions to optimize morphophysiological characteristics and stimulation of biosynthetic pathways of secondary metabolites in A. othonianum under in vitro conditions. Therefore, the work was divided into two tests, 1 - Determination of the best light intensity (12, 25, 50, 100 and 150 μmol m-2 s -1 ) and 2 - Determination of the best spectral quality, using LEDs (white, blue, green, red and blue + 1: 1 red). In each experiment the nodal segments were cultivated in 250 mL flasks with 50% MS medium containing agar, with 30 μM benzylaminopurine (BAP), 3% sucrose added under 16h photoperiod. After 50 days of treatment imposition, the seedlings were submitted to pigment, photosynthetic efficiency, biometrics, anatomical, enzymatic and chromatographic analyzes. This paper is the first report with physiological approach on the influence of light intensity and quality on A. othonianum in vitro. It was found that irradiance around 100 μmol m-2 s -1 under white light or blue / red association are the most satisfactory for the maintenance of plant homeostasis, the stimulation of photoautotrophism and the activation of biosynthesis pathways phenolic compounds.
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    Patente de invenção e acesso a medicamentos em tecnologias sanitárias de alto custo
    (Universidade Federal de Goiás, 2019-12-10) Santos, Caroline Regina dos; Lima, Jordão Horácio da Silva;; Santos, Nivaldo dos;; Santos, Nivaldo dos; Moura, Katia Karina Verolli de Oliveira; Tarrega, Maria Cristina Vidotte Blanco; Lima, Jordão Horácio da Silva; Santos Junior, Clodoaldo Moreira dos
    The object of this study is the access with high cost technological drug for the Brazilian population. The general objective is to analyze whether the invention patent is in addressing the challenges for access to medicines in high cost health technologies for the realization of the constitutional right of health from the perspective of pharmaceutical care. The specific objectives are to provide historical and theoretical knowledge to present the historical evolution of international intellectual property regimes, the interrelationship with Brazilian domestic law, to demonstrate what the international scenario was like before before the TRIPS agreement and analyzing the compromising impacts related to innovation bring the consequent problem of access to medicines. Qualiquantitative medotology is used for a broader coverage of the theme, with the inductive method to compare the geopolitical behavior of the countries India, China and Brazil, focusing after the TRIPS Agreement and the deductive in the theoretical discussions. The justification is that public health is a worldwide problem, especially in developing countries such as Brazil, because there is a lack of investments in research for the quality and life of the population. It is also justified because it shows that the emergence of neglected diseases typical of developing countries creates a promising scenario for the creation of an international treaty between public and private entities for access to this neglected population. The key question is what are the real factors that drive the pharmaceutical industry to invest in research, development and innovation in developing countries? Low investment in R & D & I leads to a gap between technological innovation in developing countries. So far, the partial results show the legal milestones of innovation in Brazil, showing that R & D & I is growing slowly, but the legalistic scenario is more propelling than in the last decade, bringing tax benefits to industries that commit to investing in the environment. drug market for neglected diseases. The pricing of medicines is directly proportional to the access to medicines that are granted via administrative or judicial. Besides all the problematization in the incentive of R&D&I, there is bureaucracy of the regulatory agencies that effect the access to medicine like Anvisa. The high cost of high-tech medicines impacts the public coffers, as well as the health plans that are triggered. There is a risk sharing agreement with the pharmaceutical industry in the negotiation for access to medicines, and in some cases it is up to the courts to settle conflicts of interest.
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    Estudo da formação de biofilmes bacterianos em endopróteses (“stent”) e inibição da formação de biofilme por peptídeos antimicrobianos de venenos de artrópodes
    (Universidade Federal de Goiás, 2019-11-04) Neves, Rogério Coutinho das; Kipnis, André;; Kipnis, Ana Paula Junqueira;; Faria, Fabrícia Paula de; Carollo, Carlos Alexandre; Bataus, Luiz Artur Mendes; Fonseca, Simone Gonçalves da; Silva Junior, Nelson Jorge da
    The use of synthetic materials as temporary or permanent insertion in the body can result in infections associated with the colonization of these materials. The colonization of these materials can result in bacterial biofilms formation. Acinetobacter baumannii infections are difficult to treat due to biofilm formation and resistance to multiple drugs. Staphylococcus strains are able to form biofilm and to resist against antibiotics, in addition of being associated with endocarditis. Thus, in this thesis we hypothesize that hospital acquired bacteria were able to form biofilm in vascular stent. In this study, the biofilm formation in polystyrene plates and on coronary stents of three isolates of multidrug-resistant A. baumannii (MDR) AB 02, AB 53 and AB 72 were evaluated. The biofilm formation in coronary stents were analyzed by scanning electron microscopy. The antimicrobial peptides from wasps (Agelaia-MPI, Polydia-MPII, Polydim-I) and scorpion (Con10 and NDBP 5.8) were used to determine the minimum inhibitory concentration (MIC) and biofilm eradication (MBEC) of A. baumannii on polystyrene plates. Additionally, methicillin resistant Staphylococcus spp was also used to evaluate the ability of peptides to inhibit biofilm formation. A. baumannii MDR in contact with the vascular stent adhered to the biomaterial and initiated the formation of bacterial biofilm. The MIC of the wasp peptides against strains of A. baumannii MDR ranged from 3.12 to 6.25 μM and those of scorpion from 6.25 to 25 μM. The MBEC of Agelaia-MPI and Polybia-MPII was 6.25 μM. Con10 presented a 6.25 μM MBEC for the AB 72 strain and 12.5 μM for the AB 02 and AB 53 strains and the NBDP 5.8 inhibited at the 25 μM concentration. Agelaia-MPI inhibited the biofilm dispersion of AB 02 and AB 53 at 6.25 μM concentration and of AB 72 at 12.5 μM. Polybia-MPII inhibited at the concentration 6.25 μM. Agelaia-MPI and Polybia-MPII inhibited mature biofilms at 6.25 μM. Because Agelaia and Polybia peptides presented the best inhibitory performance they were tested against Staphylococcus ssp. Polybia-MPII and Agelaia-MPI showed MIC and MBEC of 12.5 μM. We conclude that A. baumannii forms biofilm in both plates and on vascular stents. The AMPs from wasp venoms (Agelaia-MPI and Polydia-MPII) prevent the biofilm formation of both A. baumannii and Staphylococcus epidermidis. These peptides were also able to reduce the bacterial load of biofilm-containing biomaterials. Therefore, we suggest that the Agelaia-MPI and Polybia-MPII antimicrobial peptides may be modified to coat biomaterials and prevent biofilm formation as well as for the treatment of individuals afflicted with contaminated biomaterials.
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    Influência de variantes polimórficas de CYP2C19 na aterosclerose
    (Universidade Federal de Goiás, 2019-09-04) Costa, Iasmim Ribeiro da; Moura, Kátia Karina Verolli de Oliveira;; Moura, Kátia Karina Verolli de Oliveira; Cruz, Aparecido Divino da; Costa, Sérgio Henrique Nascente; Reis, Paulo Roberto de Melo; Silva, Rita de Cássia Pereira da Costa e
    The cytochrome P450 (CYP) family enzymes are responsible for detoxification of the organism, as well as acting on the biotransformation of various drugs. Atherosclerosis patients undergoing interventional procedures are at increased risk for thrombus formation, and the use of platelet antiaggregants is required. Clopidogrel antiplatelet is a prodrug that needs to be activated by CYP family enzymes. Polymorphisms of this family, besides being related to atherogenesis, may influence the response to this drug. This study aimed to verify the possible association of CYP2C19 genotypes in response to clopidogrel. Two hundred and ninety-nine DNA samples from patients with and without atherosclerosis for the CYP2C19 * 2, * 3 and * 17 polymorphism were analyzed by ARMS-PCR and PCR-RFLP techniques. Regarding the atherogenesis process, the presence of two polymorphic * 2 alleles or one * 3 allele may have influenced the development of atherosclerosis. Allele 17 functioned as a protective factor, even in the presence of a * 2 or * 3. The combination of two mutated * 2 alleles and the presence of * 3 influenced the development of atherosclerosis, even when * 17 was present. By evaluating CYP2C19 polymorphisms and their association with disease progression, the need for stenting, generated somewhat contradictory results. Regarding response to clopidogrel and combinations of CYP2C19 polymorphisms, we found that the poor metabolizing phenotype (* 1 * 2 / * 1 * 3 / * 1 / * 17) was more prevalent in the restenosis group, reflecting the lack of activation of clopidogrel by the enzyme CYP. Although there are contradictory studies regarding the response to clopidogrel in patients with loss of function polymorphisms for the CYP2C19 gene, we found that allele 2 not only influences the atherogenesis process, but also influences the response to clopodigrel.
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    Caracterização citogenética de bovinos da raça Nelore (Bos taurus indicus, Linnaeus 1758)
    (Universidade Federal de Goiás, 2019-09-13) Amancio, Andréia Pires; Cruz, Aparecido Divino da;; Cruz, Alex Silva da; Amaral, Alliny das Graças; Cruz, Aparecido Divino da; Silva, Claudio Carlos da; Brito, Cintia Pelegrineti Targueta de Azevedo
    The agricultural sector has been standing out in the Brazilian economy in recent decades for significant increase in productivity and its growing importance for maintaining the country's balance of trade. After the introduction of cattle in Brazil and their subsequent dispersal throughout the national territory, there was an intense process of adaptation of certain groups to specific environmental conditions of each place or region, which is responsible for the formation of several local Brazilian cattle breeds. These animals may present numerical and/or structural chromosomal alterations, which may result in a group of undesired characteristics to the producer that intends to expand his herd or even for the preservation of characteristics of a species. The aim of this study was to standardize the C, GTG and NOR banding for the bovine subspecies (Bos taurus indicus, Linnaeus 1758) and subsequently produce a pancentromeric probe by amplification and labeling the centromeric regions of autosomal chromosomes of cattle. Bovine blood samples were collected to detect metaphasic chromosomes using the cell culture method. Subsequently, cytogenetic banding, amplification, chromosomal probe labeling and FISH technique were performed. Through the banding it was possible the correct chromosomal pairing of these animals and with the use of the pancentromeric probe, the marking of the centromere regions of all autosomal chromosomes. These results may offer a new tool that meets the needs of genetic improvement of these animals.
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    Investigação das atividades angiogênica, antiangiogênica, mutagênica e antimutagênica de uma bischalcona AB7 sintética
    (Universidade Federal de Goiás, 2018-08-20) Pontes, Susy Ricardo Lemes; Macedo, Maria Lígia Rodrigues;; Silva Júnior, Nelson Jorge da;; Silva Júnior, Nelson Jorge da; Telles, Mariana Pires de Campos; Saddi, Vera; Reis, Paulo Roberto de Melo
    Plant biodiversity is considered the main source of secondary metabolites useful for chemical synthesis. Among the different classes of plant metabolites with therapeutic properties are chalcones. This group represents an important class of organic compounds with numerous biological activities, such as anti-inflammatory, anti-HIV, antibacterial, antifungal and anticancer. This study aimed to investigate the angiogenic, antiangiogenic, mutagenic and antimutagenic potential of bischalcone synthetic αβ7E, 4E-1- (4-chlorophenyl) -5- (2,6,6-trimethylcyclohexen-1-yl) penta-1,4-dien -3-one (CAB7β), besides its optical properties. The angiogenesis test with the chorioallantoic membrane (CAM) of embryonated chicken eggs (Gallus domesticus) with the S180 sarcoma tumor line, the Ames and micronucleus mutagenicity tests in mouse bone marrow, and the techniques spectroscopic two-photon absorption (2PA), visible ultraviolet (UV-vis) and Z-Scan, respectively. In the angiogenic test, CAB7β promoted a significant reduction (p <0.05) in the total vessels, junctions, length and caliber of the blood vessels stimulated by the S180 cells, besides reducing the presence of the inflammatory elements in the vessels of the CAMs. Another observation was that MCAs treated with CAB7β only showed significant reductions (p <0.05) in the angiogenic process compared to the negative control group used in this study. In the Ames test, CAB7β showed a protective effect on DNA against the mutagenic action of 4-nitroquinoline-1-oxide and sodium azide, and also against the action of doxorubicin in co, pre and post-treatment models using the micronucleus test. CAB7β at the doses of 12.5 and 50 μg / plate presented a moderate cytotoxic profile in TA98 strain of S. typhimurium. In the micronucleus test, its higher dose (50mg / kg p.c.) also exhibited cytotoxic activity. However, an anti-cytotoxic pertituent of this compound was observed at all doses tested in TA100 and TA-98 strains of S. typhimurium and during co, pre and post-treatment in the micronucleus test. The experimental and theoretical results of the optical properties of CAB7β demonstrated that the absorption spectrum of this compound has an intense absorption in the region of the UVA, which is associated to a transition of the type π → π * between the HOMO - LUMO orbitals, with a transfer intramolecular charge ratio of the trimethylcyclohexen-1-yl ring to the chlorophenyl ring. Thus, we conclude that the results of this study demonstrated, therefore, the versatility of CAB7β. This molecule may be a promising target in the development of new therapies against cancer, especially against sarcoma, due to the antiangiogenic potential associated with the reduction of inflammatory elements and their significant anti-mutagenic and anti-cytotoxic effects. In addition, to present potential for the development of photoprotective filters and in photodynamic therapy given their optical properties.
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    Aumento na taxa de diagnóstico genético dos pacientes a partir da identificação de CNVs, por CMA, envolvendo genes implicados com a manifestação clínica da deficiência intelectual
    (Universidade Federal de Goiás, 2019-05-27) Pinto, Irene Plaza; Pogue, Robert Edward;; Cruz, Aparecido Divino da;; Cruz, Aparecido Divino da; Silva, Cláudio Carlos da; Minasi, Lysa Bernardes; Moura, Katia Karina Verolli de Oliveira; Brasil, Maria das Graças Nunes
    Intellectual disability (ID) is characterized by significant impairment in both cognitive and adaptive functions, originating before the age of 18 years. In addition, it is a common phenotype sign in a cluster of heterogeneous syndromic or non-syndromic disorders, associated with some comorbidities such as autism and congenital malformations. In the worldwide, ID affects around 1–3% of the general population and in Brazil ID affects approximately 0.8% of the population. The Copy number variations account for about 15– 20% of children with unexplained ID, compromising the functioning of several genes, with more than 1,416 genes described as causative of this phenotype sign. The aim of the study was to evaluate the occurrence of CNVs, identified by CMA with the size filter of < 100 kb, harboring genes functionally associated with ID in patients from SUS with a clinical diagnosis of ID referred for the genetic diagnosis. During January 2013 to December 2016, GTG banding karyotype was performed in 325 patients with ID, achieve the genetic diagnostic in 57.2%, demonstrating to be an important screening approach for patients with DI. However, 42,8% of the patients showed the karyotype with no visible numerical or structural alterations. The CMA analysis with the size filter of ≥ 100 kb was performed in these patients, where it was possible to elucidate the genetic diagnose in 29.8% of the patients, demonstrating 7,1 % of the increment on the diagnostic. All the cases remained without a diagnosis were submitted to the CMA analysis with size filter of < 100 kb, where it was identified loss CNVs in regions harboring CNTNAP2, FGF13, MID1, MID2, SHANK3, IL1RAPL1, DMD, and PAK3 genes. The reduction of the size filter demonstrated an increase of 12% in the ratio of diagnosis, expanding the spectrum of CNVs identification in regions which harboring genes related to the clinical manifestation of ID. The application of both GTG banding and CMA with the size filter of ≥ 100 kb and later the size filter of < 100 kb allowed an increase in the genetic diagnosis of ID and comorbidities, giving a broad understanding of the genetic aspects related to these conditions and allowing the adequate management of families. Finally, the genetic counseling provides a better understanding of the genetic causes of ID, the familial implications of the genetic contribution and the chance of recurrence.
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    Potencial biotecnológico de linhagens bacterianas endofíticas de Anacardium othonianum Rizzini
    (Universidade Federal de Goiás, 2017-09-29) Faria, Paula Sperotto Alberto; Martins, Paula Fabiane;; Sales, Juliana de Fátima;; Sales, Juliana de Fátima; Martins, Paula Fabiane; Pinto, Gustavo Adolfo; Gonçalves, Priscila Jane Romano de Oliveira; Vitorino, Luciana
    Plants are filled with microorganisms that colonize their structures, those capable of inhabiting plant tissues internally are known as endophytes and provide new nutritional and defense pathways, acting as a plant strategy to withstand biotic and abiotic adversities. Anacardium othonianum Rizzini, also known as cerrado-tree-cashew, is one of the main cashew trees of economic and social importance in the central-western region of Brazil, but its endophytic bacterial community has not been investigated yet. In this regard, the aim of this work was to define endophytic bacterial lineages with functional potential to promote growth of Anacardium othonianum Rizzini. Root fragments of A. othonianum were inoculated in a culture medium. Grown colonies were isolated, purified and the identification was performed by comparing the 16S rDNA sequence in the GenBank database. Subsequently, in vitro tests of biostimulation, biofertilization and biocontrol were performed to select lineages with multifactors that promote plant growth. Two endophytic lines were selected and investigated with in vivo inoculation in A. othonianum to confirm their growth promoting action. This work is the first report on the isolation, selection and application of endophytic bacteria that promote plant growth of A. othonianum. Twenty-two species were isolated, of which Acinetobacter sp. Bac109, Bacillus mycoides Bac160, Lysinibacillus sphaericus Bac161 and Pantoea agglomerans Bac131 with multifactors of interest. Co-inoculation of A. othonianum with the endophytic bacteria Acinetobacter sp. (Bac109) and Pantoea agglomerans (Bac131), promoted an increase in initial growth.
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    Uso de nanopartículas metálicas na vacinologia: implicações para o desenvolvimento de vacinas contra doenças infecciosas
    (Universidade Federal de Goiás, 2018-10-09) Marques Neto, Lázaro Moreira; Kipnis, André;; Kipnis, Ana Paula Junqueira;; Kipnis, Ana Paula Junqueira; Guilo, Lídia Andreu; Campos, Helioswilton Sales de; Fonseca, Simone Gonçalves da; Silva, Roosevelt Alves da
    The search for new adjuvants is the main goal in vaccinology. Along with this, understanding the impact of using nanoparticles as a delivery system and immunomodulator in vaccine systems directly impacts the development of new vaccines. In this work, we seek to study and elucidate the adjuvanticity of magnetic nanoparticles, as well as its immunogenicity and protection of the vaccine systems. Initially, a literature review was made seeking scientific bases that demonstrated the possibility of using metallic nanoparticles (MeNPs) as innate immune system stimulators. It was also sought to find elements in which metallic nanoparticles could aid in the generation Th1, Th17 and T CD8 type cellular response. From this review, it was verified that the magnetic nanoparticles, or with metallic ions, were able to stimulate the activation of costimulatory molecules (CD80, CD40 and CD86), to induce secretion of cytokines (IL-1, IL-6, IFN-γ and TNF-α) as well as the humoral immune response, but no work demonstrated whether these nanoparticles were able to induce cellular response. Consequently, in the second part of the study, tuberculosis was used as model to verify if a vaccine formulation with a magnetic nanoparticle of manganese ferrite combined with recombinant fusion protein would have the ability to induce a protective cellular immune response, without adding other adjuvants. The nanoparticle was coated with recombinant CMX fusion protein and BALB/c mice were vaccinated with this formulation, in protocol with three vaccinations with 21-day intervals. Subsequently, the vaccinated animals were infected with Mycobacterium tuberculosis (H37Rv) to evaluate the protection conferred by the vaccine. The results showed that the nanoparticle was able to generate cellular immune responses of Th1, Th17 and T CD8 types, depending on the route of inoculation (subcutaneous, intranasal and mixed). The most preeminent response was Tc1 which was recalled after infection was able to protect against the challenge with Mtb. In addition, there was no appearance of side effects or damage to organs of infected animals, demonstrating that the formulation is safe. Finally, the vaccine formulations with MeNPs, more specifically with manganese ferrite, demonstrate potential application in vaccinology, and may be applied in vaccine formulations to generate cellular immune response, but the route must be considered and in case of use other adjuvants it should consider the possible interaction of NP with the molecule and their ligand.
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    Nanocompósitos biocompatíveis de poliuretana com hidroxiapatita e nanocelulose
    (Universidade Federal de Goiás, 2018-04-02) Arantes, Thaís Moraes; Alcantara , Glaucia Braz;; Lião, Luciano Morais;; Lião , Luciano Morais; Camargo, Emerson Rodrigues de; Rabelo, Denílson; Oliveira, Emília Lima de; Castro, Carlos Frederico de Souza
    Polyurethane nanocomposites were prepared with nanoparticles of hydroxyapatite and/or nanocellulose in order to obtain biocompatible materials. The nanocelluloses were obtained from sugarcane bagasse, seeking to evaluate their use as a source of cellulose nanofibers. The conditions of extraction and separation of the nanocelluloses were investigated. The effect of time and concentration of sulfuric acid was studied, resulting in particles with reduced size and homogeneity in the size distribution without cellulose degradation. The cellulose nanocrystals that presented the best results regarding suspension staining, crystallinity index, thermal properties, particle size and Zeta potential were those hydrolyzed in 50% H2SO4 at 45 ° C for 2 hours. For the synthesis of the hydroxyapatite (HA) nanoparticles, a multivariate statistical analysis was carried out using a factorial design with resolution 23 . From which an empirical model was created that allows the control of the shape and size of the hydroxyapatite nanoparticles. HA nanoparticles with sizes varying from 8 nm to 600 nm were formed by oriented coalescence growth mechanism. The structure was confirmed by images of Electron Transmission Electron Microscopy and Scanning Electron Microscopy. The hydroxyapatite nanoparticles presented a well-defined nanorod shape with a narrow size distribution. It was observed that the model was statistically significant and the main parameter for the growth of crystals in the hydrothermal process was the temperature. Polyurethanes derived from castor oil were synthesized without residues of their monomers. The insertion of the nanoparticles into the polymer matrix improved the thermal stability of the composite. Finally, the cell viability assay showed that polyurethane nanocomposites with hydroxyapatite are biocompatible and in this way can be used as biomaterial.
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    Desenvolvimento de um sistema multiplex de loci STRs autossômicos polimórficos para a identificação humana
    (Universidade Federal de Goiás, 2017-09-05) Rodovalho, Ricardo Goulart; Pereira, Rinaldo Wellerson;; Cruz, Aparecido Divino da;; Cruz, Aparecido Divino da; Vieira, Thaís Cidália; Rodrigues, Flávia Melo; Moura, Kátia Karina Verolli de Oliveira; Junior, Walter Pinto
    The main scope of the current study was to develop a short tandem repeat (STR) multiplex system, made up of 22 highly informative loci, for application in forensic genetics. The system comprised of 21 polymorphic autosomal short tandem repeat loci, namely D3S1358, TH01, D21S11, D18S51, Penta E, D5S818, D13S317, D7S820, D16S539, CSF1PO, Penta D, vWA, D8S1179, TPOX, FGA, D2S441, D17S1301, D19S433, D18S853, D20S482 and D14S1434, and the amelogenin gene locus. Strategies were developed to overcome the challenges involved in creating a multiplex system. Based on the literature and available databases, STR loci were selected to obtain discriminatory markers, and followed specific criteria for this purpose. Primers were designed using the Primer3 software and the AutoDimer was used to evaluate potential interactions between them. The 21 selected STR loci were validated individually and jointly, both to assess their sensitivity and to test the efficiency of the multiplex system. Statistical analyses were based on the genetic data of 450 unrelated individuals living in the State of Goiás, thus allowing the establishment of the parameters necessary to use this system. A total of 239 alleles were detected for the 21 loci in the set, allowing for a probability of identity of 4.23 x 10-25 to be obtained. The combined power of discrimination was 0.999999999999999999999999 and the combined power of exclusion was 0.99999. Upon complete validation of the entire system, this multiplex assay was considered to be a powerful tool for application in human identification by DNA analysis.
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    Disponibilidade e mobilidade de elementos-traço em sistemas aquáticos tropicais
    (Universidade Federal de Goiás, 2017-09-27) Costa, Henrique Santana;; Zara, Luiz Fabrício;; Rocha, Cleonice;; Rocha, Cleonice; Tejerina-Garro, Francisco Leonardo; Campos, Alfredo Borges de; Telles, Mariana Pires de Campos; Longhin, Sandra Regina
    The complexity of an aquatic system requires understanding the dynamics of its components - both spatially and temporally. The trace elements, being found under different oxidation states (due to their high enrichment factors and low removal rates) are an important environmental stressor. This research aims to determine the availability and mobility of trace elements (Zn), cadmium (Cd), lead (Pb), copper (Cu) and chromium (Cr) between water, sediments and fish in rivers located in the Paraná river basin, in the state of Goiás, Brazil. In order to compare the distribution of these compounds and the dynamics of their flows between fish, sediments and water, data were collected during the dry season (April to September), using descriptive and multivariate statistics data analysis. The analysis of the distribution coefficient (K d ) of the trace elements Cr, Cu and Zn suggests that the water-sediment interaction was influenced by environmental variables: the trace element Cu was influenced by the water temperature; the trace element Zn, in turn, was influenced by the pH of the water. As regards the biotic component, the analysis of the distribution coefficient (K d ) of the trace elements Cr, Cu and Zn (obtained from muscle tissue samples of the fish species collected) suggests that the biomass, migratory habit and basin (hydrology) of the rivers sampled influenced the bioaccumulation of trace elements by fish species. On the other hand, the variables group-trophic, lithology and vegetation cover did not influence the bioaccumulation of trace elements by fish species. It was not observed a relation between the distribution coefficient (K d ) of the trace elements Cr, Cu and Zn in the water-sediment-fish interrelationship, which suggests that the presence/incorporation of these metals occurred independently. There was no indication that geological variables (lithology), anthropic (removal of riparian forest), chemical (pH), hydrological (water velocity) and physical (water temperature) variables influenced the interrelation of Cr, Cu and Zn between analyzed components. The steps involved in this study, as well as the results obtained, characterize the importance of research aimed at a better understanding of the processes that control the availability and mobility of trace elements in aquatic systems.
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    Efeitos do tratamento com a Hev b 13, extraída do látex natural de Hevea brasilienis, na resposta inflamatória de ratos com sepse
    (Universidade Federal de Goiás, 2017-03-06) Araújo, Lilhian Alves de; Mrué, Fátima;; Silva Júnior, Nelson Jorge da;; Silva Júnior, Nelson Jorge da; Reis, Paulo Roberto de Melo; Gomes, Clayson Moura; Pelli, Milton Adriano de; Silva, Cláudio Carlos da
    Sepsis is an infectious disease characterized by severe systemic inflammatory response. Rupture of the complex equilibrium between inflammatory mediators in the acute phase of the disease leads to exacerbated production of proinflammatory cytokines, with consequent hypotension, increased capillary permeability, organ lesions and death. Recent research using the Hev b 13 protein derived from natural Hevea brasiliensis (rubber tree) latex has demonstrated important anti-inflammatory and immunomodulating effects. The aim of this study was to investigate the effects of Hev b 13 on the systemic and tissue inflammatory response of septic rats. To that end, cecal ligation and puncture (CLP) was performed on male Wistar rats and after six hours, the animals were randomized into groups and subcutaneously treated with doses of 0.5; 2.0 and 3.0 mg/Kg of Hev b 13. Next, the animals were subdivided into three different times (1, 6 and 24 hours after treatment) for blood sample collection and euthanasia with removal of the lungs and liver. Leukocytes, tumor necrosis factor- alpha (TNF-α) plasma and tissue levels, interleukin (IL)-6, IL-10, and IL-4 and histological slides were analyzed. The results demonstrated that treatment with the Hev b 13 protein prompted a significant decline in total and differential leukocytes, as well as the production of TNF-α and IL-6, associated with an increase in IL-10 and IL-4 in plasma and lung tissue. Moreover, it restricted the morphopathological changes found in the lungs, including neutrophil infiltration, swelling and alveolar thickening. In the liver, it increased IL-10 production and inhibited TNF-α and IL-6, in addition to reducing hemorrhage, sinusoidal inflammatory infiltrates and hydropic degeneration in histological assessment. We conclude that Hev b 13 displays anti-inflammatory and immunomodulating activity capable of attenuating lung and liver lesions in rats during acute sepsis, with potential for clinical applications.
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    Avanços tecnológicos e variabilidade genética da expansão CGG da região promotora do gene FMR1
    (Universidade Federal de Goiás, 2016-02-02) Gigonzac, Marc Alexandre Duarte; Pereira, Rinaldo Wellerson;; Cruz, Aparecido Divino da;; Cruz , Aparecido Divino da; Ayres, Flavio Monteiro; Oliveira , Kátia Karina Verolli de; Silva, Claudio Carlos da; Reis, Angela Adamski da Silva
    X-Fragile Syndrome (FXS) is the leading cause of inherited intellectual disability in the world and the second of genetic etiology, with an estimated prevalence of 1/4000 men and 1/8000 women. The most common molecular mechanism in SXF is due to changes in the expression of the FMR1 gene, located in Xq27.3, due to CGG trinucleotide expansions in the promoter region and subsequent methylation of the gene. In spite of presenting consistent clinical findings, they are not exclusive, and the existence of carriers of alteration in the FMR1 gene without apparent clinical manifestations makes it impossible to diagnose SXF based only on the evaluation. In the present study, a methodological proposal for the molecular diagnosis of X-Fragile Syndrome was developed from the methylation-specific triple amplification of the promoter region of the FMR1 gene combined with capillary electrophoresis. Thirty-four patients with clinical indication of SXF were referred to a laboratory of the public health network. After extraction and quantification of the DNA, the samples were amplified in an optimized protocol and the products submitted to 36cm capillary electrophoresis to verify the amount of CGG repeats and the degree of DNA methylation of each sample. Pre-mutation (3%) and six complete mutations (18%) were detected, all of which revealed a high degree of methylation. Considering the clinical signs commonly presented, the patients were also analyzed for the occurrence of Autism Spectrum Disorder (ASD), which shadowing and overlapping the SXF, verifying that 100% of the individuals with complete mutation presented the phenotype. Thus, it was possible to observe small behavioral differences in the patients analyzed, indicating a lighter clinical picture regarding aspects of social interaction and stereotypies. Thus, the new methodological proposal allows to effectively determine the CGG trinucleotide expansions in FMR1 allowing an assertive diagnosis of SXF for the families of patients attended in the public health network in Goiás.
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    Regulamentação jurídica da nanotecnologia
    (Universidade Federal de Goiás, 2016-09-12) Nolasco, Loreci Gottschalk; Martines, Marco Antonio Utrera;; Santos, Nivaldo dos;; Santos, Nivaldo dos; Neves, Cleuler Barbosa das; Araújo, Luciene Martins de; Tarrega, Maria Cristina Vidotte Blanco; Pereira, Zefa Valdivina
    INTRODUCTION. Research on the nanotechnology jobs emerge as one of the most spectacular possibilities of science in the twenty-first century. With skills of building materials, devices and systems with atomic precision, nanotechnology promises to improve human skills, new industries and products, social outcomes and quality of life, with the potential to produce considerable economic-political-social-environmental and legal impacts. It is estimated that by 2020 will be moved globally about $ 3 trillion, with about 20% of all manufactured products in the world based to some extent, the use of nanotechnology and that all semicondutores sector and half of the pharmaceutical industry relies upon new materials; besides directly involved six million jobs. This is due to its incorporation into various existing technologies (physics, chemistry, biology, medicine, electronics etc.), the result of technological convergence with ability to create new forms of life, but with the possibility to increase the risk of unlimited and unmanageable, which would, if released, interact differently with living systems, causing surprising effects and unpredicted (which is not provided) or unpredictable (which is unable to be provided), and potentially more toxic than the same mass equivalent conventional and larger particles. Given the unpredictability of its results due to scientific uncertainties about the understanding of the risks related to the small size, area and surface chemistry, solubility and size, nanoparticles could cause disturbances in molecular and cellular levels. Similar in size to biological macromolecules such as proteins, DNA and phospholipids, have important consequences and can lift entirely unprecedented ethical principles in relation to other biotechnologies. Studies have reported potential toxicological effects of nanoparticles on human health result of interactions and biological, physical and chemical changes in various organic functional systems such as respiratory, digestive, nervous, lymphatic, excretory, blood circulation, skin, breast milk, muscle and placenta; as well as contamination of the environment. In the specific case study on the use of titanium dioxide nanoparticles in sunscreens, it was concluded by exposing both in professional environments or between the population groups and living organisms (biota), almost all phases of the lifecycle, and no exposure pathway can be dismissed as irrelevant to the workers. Nevertheless, is available for sale a quantity greater than 1,800 products and services of the most diverse, including medicines, cosmetics and foods containing nanomaterials and nanoparticles, and may expose the health and safety of consumers and workers globally to your process manufacturing and marketing. Because nanotechnology be able to act in a fundamentally different way compared to their respective material macro scale, it has been impossible to infer the safety of nanomaterials using the information derived from the bulk source material. In Brazil and almost the entire globe, there is no specific legislation with requirements of new and specific methods and assessment tools when a compound product of larger scale is replaced with the same compound nanoscale, getting health and safety aspects, and ethical, social and governance issues, short of nanotechnology development. Although there is no specific regulatory framework for the area where the products are registered in different countries, including Brazil, the respective regulatory agencies do it according to its type, in case-by-case basis, using normative applied generally to the chemicals, pharmaceuticals and cosmetics, even without any explicit reference to nanomaterials. Representatives of various organizations, governmental or not, and scientific groups, national and international express doubts about the ability of regulatory legislation, research standards and methods and conventional measurement strategies of non-specific toxicity applied to nanotechnology. GOALS. The specific aim of the thesis was to investigate whether the Brazilian legal system is (in) sufficient to specifically meet the technological innovations inherent in nanotechnology, with demands for creation of methodologies to identify, evaluate and manage the possible risks throughout the life cycle of nanomaterials and nanoparticles through prevention and precautionary instruments before placing on the market of products, services and processes that contain nanotechnology. It is understood that the legal regulations should also require the adoption of proper disposal of waste production measures, and to establish procedures for civil liability, criminal and administrative those involved, if applicable. The purpose of the legal regulation is to provide legal certainty for consumers and employees of present and future generations (prospective focus) and the environment. METHODOLOGY. The study had the scope to carry out exploratory and bibliographic research through survey data in the literature. Literature searches were performed by databases CAPES, is consulting with original and review articles on the subject Nanotechnology, Risk, Regulation; as well as specific books of Nanotechnology Area and Law. We also used the analytical method in the study of nanotechnology risks, the prospective liability, the precautionary principle and the analysis of the legislation. Based on the 1988 Federal Constitution, under the paradigm of democratic rule of law, the survey adopted the garantista theory and substantialist of law, which is based on the guarantee and direct application of fundamental rights, indicating that economic agents, companies and state should pay attention the mandamentais precepts of the Constitutional Charter and the infra-constitutional legislation, to at least achieve a standard of conduct that meets the dictates of fundamental rights, so it is possible to observe that economic development is not more important than human development, both one and the other, promises of converging technologies. RESULTS AND CONCLUSIONS. The advance of science, particularly in the sector of biology, genetic engineering, chemistry, medicine, biotechnology and nanotechnology, have imposed and continue to impose the state and the law, increased vigilance for possible risks and perpetráveis damage to physical and mental integrity human beings, so that scientific progress reconciles with the standards and tutelary principles of human personality, recognized and established in the 1988 Constitution, considered in the current democratic state paradigm of law, the center of the legal system. The Law as a science, through the establishment of legal instruments must create preventive and precautionary management measures for risk, based on the constitutional principles of information and accountability, the one that underlies the principle of focused caution for a time span (prospective) so far disregarded the law. In practice, in legal and constitutional terms, it implies the obligation to adopt appropriate safety and precautionary measures ordered and anticipatory (legislation, assessment tools and risk management), which limit or neutralize the causation of damage with a total irreversibility or generates partial effects negatively disturbing damage and imbalances of the decent survival of human life and all forms of life centered on the balance and stability of natural ecosystems or processed. Currently innovation in nanotechnology applications is proceeding ahead of regulatory policy, raising concerns that ethical, economic, legal, social, toxicological and environmental issues are delayed or lagged. These concerns challenge, globally, governments, manufacturers and civil organizations to establish a legal and judicial system that addresses new methods of management and monitoring of probable and some proven risks and damage during the production chain and post-marketing products and nanoparticles containing nanomaterials. New approaches must incorporate criteria of size, shape, surface area, activity and structure area, and require the construction of new detection tools, monitoring and adequate characterization of nanomaterials, as well as the understanding of processes occurring on the surface of nanoparticle when in contact with living systems in order to understand the possible toxicological effects, and therefore address the specificities of control and risk management throughout the production chain and life cycle of products and services with nanoparticles. Although exposures of workers, consumers and ecosystems contact applications and products containing nanomaterials are subject to a significant gravity context, putting workers at risk of exposure through inhalation, skin absorption or ingestion, and despite calls for moratorium on governments and atentassem industries to the problems generated by technologies convergence, the legal regulation of nanotechnology inched around the globe. Inobstante, concluded that reflex and partially by means of integrative interpretation of legis analogy, the Brazilian legal system provides regulations to nanotechnology, to identify responsibility, measurement of parameters, penalty and establishment of cautious conduct in dealings with the risk nanotechnology, in particular by establishing the precautionary principle, such as the Biosafety Law no. 11,105 / 2005 the National Policy on Solid Waste no. 12,305 / 2010; Nuclear Activities no. 6,453 / 1977, as well as judicial practice with the application of international treaties ratified by Brazil, before and after the enactment of the Brazilian Constitution of 1988, including, Agenda 21 (1992) and the Cartagena Protocol on Biosafety (2000) and the recourse to constitutional principles inspiring the juris analogy of the system, the principle of reasoned caution to the general safety requirement and full compensation of the damage, the preservation of human dignity, respect for life and health. Added to that, normative not legally binding (soft law) developed by economic organizations and international standardization (codes of conduct and responsibility) for the development, marketing and nanotechnology risk management, can complement the control regulation and state control. So that stakeholders in nanotechnology should take into consideration at the time committed efforts and resources in that sense some parameters to guide its activities and observe what they represent for their activity externalities of legal regulation. The research points principles and indicators that should be deployed as supervisory measures and preventive management of risks of nanotechnology and nanomaterials in order to ensure its safe development, among which stands out the precautionary approach and mandatory specific regulations. They also point out some strategic proposals for risk management, including controls in the workplace, volunteer programs and insurance. It concluded that the risk management for protection and promotion of research activity and its holders, as well as to preserve the quality of life of the human being and the environment in general, can be made at various levels, including government regulatory agencies the definition of environmental, health and safety, companies in the implementation of industrial management programs and hygiene products and insurers in the formulation of coverage policies and prices. It remains, however, the Brazilian legal challenge in specific regulatory and comprehensively nanotechnology or promote adaptation in particular the biosafety laws and solid waste for its proper disposal, involving the generation of new methodologies and protocols with a multidisciplinary approach, between principalmente chemistry, responsible for the synthesis, quantification and characterization of materials, biology and medicine, the design of the trials and interpretation of results in order to identify and evaluate systematically materials and safer alternative processes, and thus, anticipate the risks potential products and processes containing nanoparticles and nanomaterials, paying attention to the Federal Constitution of 1988 garantista bias, and head to the field of formulation, interpretation and application of laws, which binds all state powers (executive, legislative and judicial) , industries, scientists, laboratories, universities and other stakeholders in the development of nanotechnology, with purpose to ensure that the company will enjoy the economic and social benefits that nanotechnology promises widespread offer.