Mestrado em Biotecnologia

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Papel da ovariotectomia no aparecimento de sinais histopatológicos associados ao Mal de Alzheimer em camundongos APOE Knockout
(Universidade Federal do Espírito Santo, 2010-07-27) Destefani, Afrânio Côgo; Rangel, Letícia Batista Azevedo; Silva, Ian Victor; Pereira, Rodrigo Romcy; Houzel, Jean Christophe
Alzheimer's disease is a progressive degenerative disease that compromises elements of the central nervous system, associated with memory loss and dementia. Are observed in AD extracellular plaques, which consist of deposits of beta-amyloid peptide and intracellular neurofibrillary tangles, which can be associated with intermediate filaments, mainly in the hippocampus and cerebral cortex. Anamnesis and molecular studies have suggested the presence of genetic elements involved in this pathophysiology, such as APOE gene, betaamyloid and TAU beyond glycemic and lipid levels. It is known that the disease incidence increases rapidly with advancing age and Alzheimer's disease affects proportionately more women than men. It is believed that the decline in gonadal hormone levels in postmenopausal women may contribute to the observed difference between the sexes. This study aimed to determine the influence of ApoE and female sex hormones on the molecular aspects involved in the formation of senile plaques in the brain of female mice C57 and ApoE knockout using ovariectomized (OVX) as an agent mimicking menopause in women. We used 14 female mice C57 and 14 female mice ApoEKO (knockout for apolipoprotein E gene) at eight months. There were histological techniques (Cajal, Bielschowsky and Thioflavine S) for the visualization of senile plaques in brain sections. Analysis of plasma glucose, cholesterol, triglycerides, c-HDL, VLDL and LDL were also evaluated. The formation and distribution of βA, Tau protein and Tau [pS199] and BDNF were analyzed via ELISA. The results suggest that ovariectomy promoted the significant depletion of estradiol (E2) and BDNF and significantly increased levels of βA, Tau and Tau [pS199] in different biological compartments when compared to controls. In the meantime, we suggest that female sex hormones along with ApoE modulates the clearance of some molecules related to the formation of senile plaques and neurofibrillary tangles characteristic of Alzheimer's disease.
Interação do alendronato e da vitamina K no metabolismo osteomineral de ratas ovariectomizadas
(Universidade Federal do Espírito Santo, 2010-07-29) Pimentel, Fernanda Scarpatti; Rangel, Leticia Batista Azevedo; Silva, Ian Victor; Bissoli, Nazaré Souza; Miranda, Kildare Rocha de
The bone is a specialized form of connective tissue that provides support for metabolic and biomechanical throughout the body. Thus, the bone is very dynamic, with constantly renewing itself. His integrity therefore depends on the balance between the processes of formation and resorption. The loss of this balance alters both the structure of organic matrix as bone mineralization. Moreover, imbalances in bone remodeling process may result in the development of systemic skeletal diseases such as osteoporosis. The osteoporosis is a chronic progressive disease that affects millions of people around the world. Only in Brazil some 10 million people suffer from osteoporosis. The hormone estrogen deficiency induced by ovariectomy (OVX) rats, demonstrably stimulates increased bone resorption, especially in long bones and spine, mimicking what happens in postmenopausal osteoporosis. In this work we used OVX rats to investigate the interaction of alendronate (a drug widely used to treat osteoporosis) and vitamin K (VK) (recent investigations have pointed anabolic bone tissue of osteoporotic patients) in the metabolism osteomineral. This study revealed that administration of alendronate (ALE) and VK with ALE (ALE+VK) produced significant recovery in bone mineral density (BMD) in OVX rats. However, the use of VK alone did not appear to make any significant effect on BMD in OVX rats. We observed an increased excretion of urinary deoxypyridinoline (DPD), a marker of bone resorption, in OVX group, and statistically significant reduction of DPD when the animals were treated with VK, ALE, or both. There was no statistically significant difference in bone mineral content and body surface area. Was verified statistically significant difference in the thickness of compact bone in the different study groups. There was also a statistically significant reduction in wet weight and endometrium of OVX rats, demonstrating the effectiveness of ovariectomy. Therefore, the animal model used in this study efficiently mimicked estrogen deficiency induced by ovariectomy, resulting in increased bone resorption; treatment with ALE and VK+ALE increases BMD in OVX rats, while the VK alone does not produce this effect; treatment with ALE and VK reduces bone resorption in OVX rats, verified by the reduction in the excretion of DPD
Mutações na região aminoterminal do gene COL1A2 e a manifestação da osteogênese imperfeita
(Universidade Federal do Espírito Santo, 2011-04-29) Almeida, Márcio Germello de; Louro, Iuri Drumond; Paula, Flavia de; Batitucci, Maria do Carmo Pimentel; Freitas, Sílvia Regina Sampaio; Bortolini, Eliete Rabi
Osteogenesis Imperfecta (OI) is a genetic disease associated with alterations in the type I collagen molecule. OI is clinically characterized by skeletal fragility and deformity. The majority of OI cases are caused by dominant autosomal mutations on COL1A1 and COL1A2 genes, responsible for the synthesis of the type I collagen molecule. However, there are isolated cases and new cases of recessive autosomal forms of OI. Thus OI is a clinically and genetically heterogeneous group, and it justifies the importance of molecular studies related to this desease. The objective of this research was to characterize the mutation pattern of the beginning region of the COL1A2 gene of OI patients. We studied the exons 1 to 26 of the COL1A2 gene on 33 non-related patients of Vitoria-ES with OI diagnosed using PCR-SSCP and sequencing techniques. We detected an alteration on the exon 16 of a female OI with moderate form of the disease, identified as c.739 G>C (p.Gly247Arg). The clinical report of our patient differs from another one already described on literature. These results may contribute to the understanding of the disease, leading to the development of more efficient treatment methods to patients with OI.
Avaliação tóxica, citotóxica e mutagênica de arilaminonaftoquinonas sintéticas
(Universidade Federal do Espírito Santo, 2011-05-16) Freitas, Josivany Valério de; Batitucci, Maria do Carmo Pimentel; Paula, Flávia de; Morales, Maria Aparecida Marin
The search for new anticancer agents has become increasingly necessary, with a view to increased efficiency and reduced cytotoxic effect on normal tissues. In this perspective, quinones which are classified according to the aromatic system in its present structure, bearing a ring the naphthoquinones naphthalenic characteristic, have been extensively studied in recent decades due to their bioreductive properties and participation in oxidative stress. Toxicological studies for testing acute toxicity, cytotoxicity, genotoxicity and antigenotoxicity of various substances, especially when aiming at producing new medicines, are performed by bioassays that evaluate the effects of the agent tested on the cell and genetic material. We propose to evaluate the toxicity, cytotoxicity and mutagenicity of arilaminonaftoquinones 1, 2 and 3 by synthetic determination of the LD50 in mice, cytotoxicity against Artemia salina larvae and the micronucleus. The test for LD50 determining was conducted in Swiss albino mice (Mus musculus) in which mice with body mass index between 25-45g, were divided into experimental groups (n = 10) and treated separately, with arilaminonaftoquinones solution of 1, 2 and 3 DMSO/H2O in concentrations of 200, 500 and 1000 mg / kg, and with the negative control (CN-0.9% saline solution) intraperitoneally in experimental casual.The lethality test to larvae of the A. saline was performed according to the method of Meyer (Meyer et al., 1982), simplified. The samples were dissolved in DMSO solution and supplemented with artificial sea water, this solution were prepared dilutions. Metanauplius larval stage (10 units) were added to each tube containing the dilutions and the cultures were incubated for 24 hours. DMSO controls (NC) and lapachol + DMSO (CP) were included in the test. O micronucleus test was performed in vivo in bone marrow of Swiss albino mice (Mus musculus). To this end, we used young animals, males and females. For each of arilaminonaftoquinone tested the animals were randomly divided into twelve groups (3 animals of the same sex per cage), the same treatment was applied to a group of males and a group of females. For each of the six arilaminonaftoquinones groups received by gavage, Arilaminonaftoquinone [50, 100 and 200mg.kg-1 mc] dissolved in DMSO. The other groups received cisplatin (3mg.kg mc-1, ip, CP), DMSO (0.005 mL.g mc-1, CS) or 0.9% NaCl (CN). Bone marrow was collected 24 hours after treatment for preparation of slides. For each animal was calculated the number of micronucleated polychromatic erythrocytes (PCEMN) in 2000 polychromatic erythrocytes (PCE) from bone marrow cells and the number of micronucleated erythrocytes normochromatic (NCEMN) in 2000 normochromatic erythrocytes (NCE) in peripheral blood. Cytotoxicity was evaluated by means of the PCE: NCE in 200 erythrocytes (PCE + NCE) in bone marrow, per animal. Statistical analysis was performed using the software Assistat ® 7.5. Statistical significance comparing data between different treatment groups was performed by ANOVA (analysis of variance) to a criterion and the Tukey test a posteriori the 5% probability. The obtained datas by the LD50 test and the front toxicity A. Salina, showed that arilaminonaftoquinones tested are classified as harmful and cytotoxic respectively. When the results of micronucleus test results show us that the animal groups treated with different concentrations of DMSO arilaminonaftoquinonas + did not differ significantly from the CS and CN ratio on the PCE: NCE and the frequency of MN, while the CP group decreased ratio of PCE: NCE (P <0.01) and increased frequency of PCEMN (P <0.01) compared to the control group, demonstrating the sensitivity of the assay to detect genotoxicity in vivo.
Efeitos de alguns tensoativos sobre a viabilidade celular de linhagens celulares de câncer de pulmão
(Universidade Federal do Espírito Santo, 2011-08-15) Aguilar, Naidilene Chaves; Silva, Ian Victor; Silva, André Romero da; Ribeiro, Joselito Nardy
The Lung Carcinoma (LG) represents the major challenge to the global health, becoming the leading cause of death by cancer among men and women in Brazil. The natural history of the disease includes high mortality rates and aggressive evolution, often with the patient coming to the doctor when the disease is already advanced. The incidence of this disease has its peak between the ages of 55 and 65 years old. The occurrence of lung cancer is intrinsically linked to the exposure to carcinogens, so that 90% of the cases are associated with active smoking. Thus, this study evaluated the feasibility of tumor lines through tests with four types of drugs of different classes. Recently, several experiments were performed with drugs that selectively inhibit the spread of tumor cells but have no effect on primary growth (Ross et al., 1969). This communication is in agreement with experiments performed to study the effect anti-metastatic of some drugs on the tumor spread. Thus, our study experiments to use the surfactants, Triton® X-100 (Sigma), Tween® 20 (BioAgency), SDS (Vetec) and CDs (Sigma) in varying concentrations, in order to evaluate cell viability. In this study we used two tumor cell lines, the H460 and A549, respectively classified as large cell carcinoma and adenocarcinoma. The screening of the substances with potential cytotoxic effect was done by the colorimetric test using MTT (3 - (4, 5-dimethyl-2-y1) 2, 5-diphenyl tetrazolium bromide), as described by Mosmann (1983). The results showed variations in cell viability shown by the strains under study when treated with different types of drugs resulting in a viability proportional to the type and concentration of the drug. He was taken into consideration the different classes and chemical structures of the substances tested to discuss the different results and different strains. The SDS showed greater cytotoxic effect and the CD with the lowest or no effect under the same 8 conditions, Tween 20 and Triton X-100 had similar effects, with Triton X-100 getting a greater reduction in cell viability.
Efeito da metil-b-ciclodextrina e de alguns tensoativos sobre a viabilidade celular de linhagens celulares de câncer de ovário
(Universidade Federal do Espírito Santo, 2011-08-16) Gonçalves, Nahun Thiaghor Lippaus Pires; Silva, Ian Victor; Ribeiro, Joselito Nardy; Silva, Andre Romero
This study sets up a correlation between some surfactants, a derivate of cyclodextrin, cyclic oligosaccharide originated from the action of the glycosyltransferase cyclodextrin, a methyllated β-cyclodextrin and the lines of ovarian cancer A 2780 and OVCAR 3, trying to offering the possibility of using cyclodextrin associated with standard chemotherapy drugs in therapy against ovarian cancer, which is pointed as one of the main causes of death among gynecological ills. Ovarian cancer is connected with high mortality rate and has several histo-clinical subtypes, raising the number of patient diagnosed each year, therefore there are the needy of continuing the scientific research which this kind of cancer is related, once the knowledge developed so far are extremely significant, yet it still is insufficient to generate a cure. Through the cellular viability by MTT method, which uses salt of tetrazol to express quantitatively the proliferation and survival of cells, and Bradford method for standardization data, it attempts to establish the influence of surfactants and some of the derivate of cyclodextrin on and between the lines of ovarian cancer. The methyl-β-cyclodextrin and the surfactant SDS, TX100 and TW20 induced a reduction in dose-dependent mitochondrial activity in cell cultures of ovarian cancer line A 2780 and OVCAR 3. The cyclodextrin derivate has demonstrated induction of gradual reduction of the mitochondrial activity of two from eight concentrations tested (0,2% and 0,4%) with cell viability of approximately 60% in strain A 2780. In the line OVCAR 3 the methyl-β-cyclodextrin shows potential cytotoxic able to tamper 25% of cell growth in conditions of treatment is reduced in this strain when compared with strain A 2780. Due to the properties of cyclodextrins, results of comparative analyzes of cell viability, in the strains of ovarian cancer, indicates to greater possibilities in future studies, enabling the research presented here as a reference.
Análise de mutações em formas recessivas de pacientes com osteogênese imperfeita do Espírito Santo : comparação de metodologias
(Universidade Federal do Espírito Santo, 2012-02-17) Quirino, Geise de Aguiar; Paula, Flavia de; Errera, Flavia Imbroisi Valle; Bortolini, Eliete Rabi
Osteogenesis Imperfecta (OI) is a genetic desease characterized by patient s bone fragility and deformity, in which severity ranges from a barely detectable connective tissue disorder to lethality in the perinatal period. The diversity of clinical variability in patients is caused by the different location or type of mutations in one of the ten genes related with the disease. This wide clinical variability difficults the perfect clinical diagnoses, due to that the use of molecular biology techniques becomes necessary to obtain a correct diagnoses and for genotype: phenotype correlation. One of the relevant genes associated with recessive forms of OI is the LEPRE-1 gene, responsible for encoding the prolyl 3 hidroxylase 1 protein. This protein and two others are components of the complex responsible for pro-collagen alfa 1 chains 3 prolyl hydroxylation. The target of this research was to analyze the LEPRE-1 gene in eight non consanguineous patients clinically diagnosed as severe Osteogenesis Imperfecta suggestive of autossomic recessive heritage by DNA sequencing of exons 1, 3, 5, 6 and 14 of the gene. In addition, the data obtained was used to analyze the efficiency of the SSCP technique by comparing the results between screening for mutations methodologies and gene sequencing methodologies. On exon 6, for instance, a mutation in one patient was found: a heterozygose base change (c.1087A>G / p.Lys363Glu), consequently, lysine was produced instead of glutamic acid. On the other exons, there was no mutation found on the patients chosen. All the results obtained in this research were compatible with datas generated by SSCP and suggest high efficient of SSCP technique for LEPRE-1 gene to recessive cases of Osteogenesis Imperfecta
Análise da expressão dos genes relacionados à parede celular e a membrana plasmática de Saccharomyces cerevisiae em resposta a alta pressão hidrostática
(Universidade Federal do Espírito Santo, 2012-02-28) Souza, Diego Trindade de; Fernandes, Patrícia Machado Bueno; Masuda, Cláudio Akio; Guimarães, Marco César Cunegundes; Bravim, Fernanda
The yeast Saccharomyces cerevisiaehas received much attention for its applications in the biotechnology industry. However, during these processes, yeasts are exposed to many different types of stress. Knowing the mechanisms S. cerevisiaeuses to respond to these stresses is essential for obtaining higher yields in the industrial sector. High hydrostatic pressure (HHP) is a stress that exerts a broad effect on S. cerevisiae, interfering with cell membranes, cellular architecture, and various biochemical processes. In this study, we used the microarray technique to investigate the influence of HHP on genes related to the cell wall and plasma membrane. For this, cells from BT0605 strain were subjected to hydrostatic pressure of 50 MPa for 30 min, and then incubated at ambient pressure for 5, 10 and 15 min.The microarray results clearly demonstrated that of the genes involved in biogenesis of the cell wall, those which are related to protection were readily activated by high pressure, whilst the genes involved in cell wall structure required 5 minutes on the ambient pressure to be induced.
Estudo da hepatite B oculta em doadores de sangue de Vitória, Espírito Santo
(Universidade Federal do Espírito Santo, 2012-02-29) Tovar, Thais Tristão; Pereira, Fausto Edmundo Lima; Louro, Iúri Drumond; Paula, Flavia de; Sá, Ricardo Tristão
Occult hepatitis B (OHB) is defined as the presence of low levels of HBV DNA in the liver or serum of individuals testing hepatitis B surface antigen (HBsAg) negative. In most cases of OHB, sera are positive for hepatitis B core antibody. The literature contains quite a few studies on the prevalence of OHB in Brazil, as well as in the worldwide population. Such reports, often controversial, demonstrate that the OHB prevalence varies among healthy individuals or patients with diseases unrelated to the liver and patients with chronic liver disease. Despite efforts, it is necessary a better understanding of: the reasons for the persistence of low levels of HBV-DNA in the absence of detectable HBsAg, the potential risk of OHB transmission and its role in the progression and aggravation of some liver diseases. Therefore, it is interesting to know the prevalence of OHB indifferent population samples which allows de monitoring of carriers of the occult infection, followed prospectively in order to try to surprise the possible effects of the presence of low levels of HBV-DNA in these individuals. In this study we investigated the presence of Occult Hepatitis B in peripheral blood obtained from 520 healthy donors of Vitoria, Espirito Santo, with the aim of guiding policies to include or not the sensitive HBV-DNA nucleic acid amplification technique (NAT) screening in blood donations with a detection limit of 54UI/mL. In order to enable the molecular detection we had also developed a method that screens plasma samples in pools which is capable of detecting the presence of HBV in the ratio of 1:40.Through the technique of nested-PCR we found that 0,2% (1/520) had occult HBV in serum samples. Despite the low prevalence of OHB detected in the study, a considerable number of patients with occult HBV infection may not have been detected if the blood units were only tested for serological markers of HBV infection. So it is important to know the prevalence of OHB in one or more additional population groups, in order to follow up carriers of occult infection prospectively to determine possible effects of the presence of HBV-DNA in low concentrations in these individuals
Análise da expressão temporal de genes relacionados ao metabolismo de Saccharomyces cerevisiae em resposta ao estresse por alta pressão hidrostática
(Universidade Federal do Espírito Santo, 2012-02-29) Silva, Lucas Ferreira da; Fernandes, Patrícia Machado Bueno; Masuda, Cláudio Akio; Louro, Iuri Drummond
Living organisms maintain a complex internal balance, which enables them to optimise their growth and metabolism. However, fluctuations in environmental conditions can affect this balance. The yeast Saccharomyces cerevisiae is a unicellular, eukaryote model organism. The biotechnological processes it is used for include the production of bread, cheese, alcohol, drugs and pharmaceuticals. From a physiological standpoint, all these processes affect the normal operation of the cell and it is extremely important to study the molecular mechanisms that provide protection against these stressful situations. One of the models used to study stress is high hydrostatic pressure (HHP), which is known to cause morphological and metabolic changes in yeast. Several studies have demonstrated that the expression pattern of genes from yeast submitted to high hydrostatic pressure shows a complex profile, including genes related to oxidative, osmotic, and heat protection.
O papilomavírus humano em meninas de uma unidade de saúde do município de Vitória : avaliação de um potencial grupo de risco
(Universidade Federal do Espírito Santo, 2012-03-09) Sartori, Mariana Penha de Nadai; Chambô Filho, Antônio; Louro, Iuri Drumond; Errera, Flavia Imbroisi Valle; Rocha, Ricardo Cristiano Leal da
Human papillomavirus (HPV) is the most common sexually transmitted virus in the world, being a cause of genital warts (Low-Risk HPV) and cervical cancer (High-Risk HPV). The virus is found in sexually active adults and adolescents, however, it has been detected in girls victims of sexual violence or not. The worldwide incidence of HPV is increasing in girls and viral transmission modes are still not completely defined. Currently, HPV prevention is based on two available vaccines only for the age group 9-26 years old and there are no available vaccines or other prevention methods for patients under nine years old, since it is not considered a risk group by competent health authorities. There are no studies of the Espírito Santo state about the HPV prevalence in girls under 9 years old and therefore their importance is not established. Because of these reasons, the aim of this study was to detect HPV in girls under nine years old, as well as finding possible routes of transmission in order to provide information for the development of preventive practices for this risk group. A total of 43 samples extracted from girls under 9 years old were analyzed using PCR, RFLP and DNA sequencing methods for viral detection and typing. Human papillomavirus was detected in 13.9% of patients, mostly low risk genotypes. Clinical and personal evaluation suggested that girls were infected by horizontal transmission and via fomites. Due to our findings, we propose that HPV infection prevention through vaccination should be extended to girls under 9 years old, especially in specific high risk populations.
Avaliação do potencial de cicatrização do óleo essencial do fruto de aroeira-vermelha (Schinus terebinthifolius Raddi) em ratas com diabetes mellitus
(Universidade Federal do Espírito Santo, 2012-04-13) Gonçalves, Elaine Paiva Moraes; Zanini, Surama Freitas; Gava, Agata Lage; Nogueira, Breno Valentim; Pereira, Thiago de Melo Costa
Diabetes Mellitus (DM) is a metabolic disease that arises from the absence of insulin and/or an inability to exhibit in a proper manner their effects. Considering that the difficulty in healing is one of the major diabetes consequences and that the pink pepper has anti-inflammatory and healing potential, we aimed to evaluate the topical use of essential oil of Brazilian red-pepper fruit (Schinus terebinthifolius Raddi) on the healing process of lesions in rats subjected to an experimental model of diabetes type I Aloxanic. After the confirmation of diabetes, the rats were subjected to the removal of subcutaneous tissue in the dorsal region of the animal and divided n groups to the received local treatment (21 days) in control (CT), commercial oil (OC) and Brazilian red-pepper (AV). After surgery, the lesion was macroscopically and microscopically evaluated in the period of 7, 14 and 21 days. The macroscopic analysis revealed a local irritation in PP group, however the microscopic analysis showed an accelerated healing process in this group in comparison with CT and CO animals at the 7th treatment day, with no differences at the 14th and 21st day. Therefore, we can conclude that after 7 days of treatment, the Brazilian red-pepper oil has a beneficial effect on the healing process of diabetic animals.
Avaliação da ação antimutagênica da ipriflavonacontra os danos induzidos por ciclofosfamida
(Universidade Federal do Espírito Santo, 2012-05-05) Delarmelina, Juliana Macedo; Batitucci, Maria do Carmo Pimentel; Paula, Flavia de; Marin-Morales, Maria Aparecida
Ipriflavone is a synthetic isoflavone derivative from daidzein and clinically prescribed for treating and preventing osteoporosis in postmenopausal women. We investigated the potential of this drug against the cytotoxic and mutagenic effects induced by cyclophosphamide (CPA) chemotherapy, using the micronucleus assay in bone marrow erythrocytes of Swiss albino mice (Mus musculus) in vivo. To evaluate their possible mechanisms of action, performed the evaluation of antioxidant activity by DPPH assay. For in vivo testing was carried out three protocols: pretreatment, simultaneous treatment and post treatment. The ipriflavone was evaluated in three different concentrations dissolved in DMSO (1,71; 8,57 e 42,85mg.kg-1 m.c) and administered by oral via. The bone marrow was collected for the evaluation of polycromatic erythrocytes (PCE) and the ratio PCE/(PCE+NCE) (polychromatic erythrocytes / polychromatic erythrocytes + normochromatic erythrocytes). For the DPPH test were assessed five concentrations of ipriflavone (500, 250, 150, 50 e 10μg.mLˉ¹) using DPPH solution (60μM). The results of in vivo tests show that the three concentrations of ipriflavone studied significantly reduced the frequency of MNPCEs induced by CPA, in the pre-treatment protocol and demonstrated the same effect at the concentrations of 1,71 e 42,85mg.kg-1 m.c in the post-treatment. However, simultaneous treatment did not reduce the frequency of MNPCE in any of the concentrations tested. In all protocols performed, the ratio PCE/(PCE+NCE) increased. There was variation between the genders in some of the experimental groups and the evaluation of antioxidant activity of ipriflavone showed no ability to donate hydrogens, suggesting that it acts through other mechanisms, such as inactivation of the enzyme activity of cytochrome P-450.
Caracterização da fadiga a partir do processamento de sinais mioelétricos e sua utilização no diagnóstico da síndrome da fibromialgia
(Universidade Federal do Espírito Santo, 2013-01-20) Moreira, Marcelo Hubner; Silva, Ian Victor; Bastos Filho, Teodiano Freire; Fioresi, Mirian; Cunha, Fransérgio Leite da
This work aims to characterize the fatigue from myoelectric signals and use them as an aid to the diagnosis of rheumatic diseases such as Fibromyalgia. The condition for this is the analysis of muscle fatigue. Through the evaluation of myoelectric signals, the behavior of muscle in some work situations was measured, such as isotonic and isometric muscle contraction, which describes the static and dynamic motor behavior. With the myoelectric signals, digital filtering techniques were applied to mitigate the noise corrupting the myoelectric signal. Then some algorithms were implemented to detect fatigue. With that, a protocol for assessing motor response based on the condition of muscle fatigue was established. In this situation, with the working muscle, the myoelectric signal acquisition was made from surface electrodes, using a commercial acquisition system. The data were processed in MATLAB R platform; algorithms were implemented for the identification of fatigue, such as RMS, MNF, ARV, MDF and AIF. In the final result, it was found that for both isometric tasks and isotonic tasks, it is recommended the use of constant weight with 60% of MCV, using MNF and RMS indicators, which were the most consistent indicators among them.
Tributilestanho prejudica ciclo reprodutivo de ratas
(Universidade Federal do Espírito Santo, 2013-01-25) Sena, Gabriela Cavati; Graceli, Jones Bernardes; Santos, Roger Lyrio dos; Andrade, Anderson Joel Martino
Triorganotins, mainly tributyltin (TBT), are environmental contaminants, commonly used in antifouling paints for boats, which suffer bioaccumulation and thus are found in mammals and humans due to ingestion of contaminated food. The importance of TBT as environmental endocrine disrupter and consequent reproductive toxicity in different animal models is well known. However, the adverse effects upon reproductive cycle are less well understood. The potential reproductive toxicity of TBT on regular reproductive cyclicity of female rats was examined. Wistar female rats at 12 weeks of age, weighing approximately 230 g were divided in two groups: Control (treated with vehicle, 0,4% ethanol solution), and TBT (treated with tributyltin, 100 ng/kg/d). The treatment lasted 16 days. TBT changed cycle regularity (%) (of animals with regular cycle), reduced the cycle duration, the proestrus and diestrus phases, and the number of epithelial cells in the vaginal smears collected during proestrous. TBT also increased the duration of metestrus and the number of cornified cells in this phase. The weight of the ovaries and levels of estradiol in serum were decreased, and we found a significant increase in progesterone levels. Histological analysis showed apoptotic cells in corpus luteum and granulosa cells layer of ovarian follicles treated animals, with cystic follicles, high number of atretic follicles and corpus luteum. The expression of the estrogen receptor alpha was reduced on uterine and ovarian tissues on TBT group. The micronucleus test (MN), using Chinese hamster ovary cells, demonstrated a concentration-dependent mutagenic effect of TBT. The toxic potential of TBT over the reproductive cycle may be attributed to changes found in the ovarian weight, unbalanced levels of sexual female hormones, and impaired ovarian follicles development.
Expressão do gene VEGFA em tecido adiposo subcutâneo e visceral de mulheres em obesidade grau III
(Universidade Federal do Espírito Santo, 2013-02-25) Silva, Fabiano Honorato Pereira e; Errera, Flavia Imbroisi Valle; Paula, Flavia de; Costalonga, Everlayny Fiorot
Obesity is a chronic and multifactorial disease resulting from a complex interaction of genetic and environmental risk. Despite efforts to develop actions to prevent and control, the prevalence of obesity continues to grow around the world. This disease is characterized by excess of body fat accumulation in adipose tissue deposits, subcutaneous (SAT) and visceral (VAT). Adipose tissue is highly vascularized and has intense angiogenic activity. For its expansion is required to occur the development of blood vessels from pre-existing vessels. The VEGFA gene is a important pro-angiogenic factors and acts promoting vasopermeability, migration and proliferation of endothelial cells. The objectives of this study were to observe the differences in the expression of VEGFA in SAT and VAT. The objectives of this study were to observe the differences in the expression of VEGFA in SAT and VAT. In addition, we also analyzed associations with clinical parameters and gene expression of adiponectin, a anti-inflammatory adipokine associated with protection against changes in glucose tolerance. In this study was performed, using the technique of quantitative PCR in real-time, the analysis of VEGFA gene expression in SAT and VAT of 33 female patients with morbid obesity (BMI ≥ 40). The sample was divided into two groups: a group with abnormal glucose tolerance (n = 9) and a group without these abnormalities (n = 19). In the total sample, the VEGFA was expressed similarly in both deposits. However, women with abnormal glucose tolerance had reduced expression of VEGFA in VAT. Moreover, in SAT the age was negatively correlated to this gene and in VAT the VEGFA gene was positively associated with overweight and negatively associated with total cholesterol and HDL. Furthermore, consistent positive correlations were observed between VEGFA and adiponectin, both SAT and in VAT. These results are important because they help to clarify the involvement and activity of VEGFA in adipose tissue
Análise molecular do gene CRTAP através da técnica de PCR-SSCP-sequenciamento em pacientes com osteogênese imperfeita do Espírito Santo
(Universidade Federal do Espírito Santo, 2013-02-28) Almeida, Lorena Schneider; Paula, Flavia de; Errera, Flavia Imbroisi Valle; Maranduba, Carlos Magno da Costa
The Osteogenesis Imperfecta (OI) is a genetic disease characterized by structural defects of type I collagen protein or by reducing its biosynthesis causing decreased bone mass and predisposition to fractures and bone deformities. Approximately 90% of individuals with OI exhibit autosomal dominant inheritance caused by mutations in the genes COL1A1 and COL1A2. However, the number of genes linked to autosomal recessive forms of OI is increasing in the literature. The CRTAP gene was the second identified causing recessive inheritance OI. This gene has 6622 bp, seven exons and encodes a protein of 46.5 kDa. The CRTAP encoding the protein cartilage associated (CRTAP) which is part of the collagen 3-hydroxylation complex, responsible for post-translational modifications during the biosynthesis of collagen molecule. CRTAP mutations are related to severe and lethal form of the disease. The target of this research was evaluating the exons of CRTAP and its adjacent regions in OI patients from Espírito Santo thought the Single-Stranded Conformation Polymorphism (SSCP) screening of mutations and sequencing. We studied 24 patients with clinical diagnosis of OI from Hospital Infantil Nossa Senhora da Glória de Vitória, Brazil. The patients’ ages ranged from 2 to 16 years (median: 14.5). The sex proportion of the patients was 15 males and 9 females. Eleven patients have mild clinical symptoms of the disease, 5 show moderate symptoms and 9 were severe cases. The lethal OI cases were not obtained by methodological difficulties. We found the polymorphisms c.534C> T previously reported in exon 2 of the CRTAP gene in patients from sample. No pathogenic mutations were found in this study. The results of this study suggest that mutations in CRTAP are rare in ES population. These data may assist in developing more efficient methodological strategies for molecular diagnosis of OI.
Identificação de mutações que conferem resistência ao tratamento com inibidores de tirosina cinase
(Universidade Federal do Espírito Santo, 2013-03-06) Moreira, Roberta Bitencourt; Renault, Ilana Zalcberg; Louro, Iuri Drumond; Von Zeidler, Sandra Lucia Ventorin; Cappelletti, Paola Alejandra
The chronic myeloid leukemia (CML) is characterized by a cytogenetic alteration known as Philadelphia chromosome (Ph), a result of reciprocal translocation t (9; 22) (q34; q11). The resulting fusion bcr-abl gene encodes a protein with tyrosine kinase constitutive activity that deregulates signal transduction inducing proliferation, apoptosis, and cellular differentiation. The natural evolution of CML is currently changing with the development of tyrosine kinase inhibitors (TKI). Imatinib (Gleevec® , Novartis) was the first TKI approved for the treatment of CML. The eight-year update of the international randomized study of interferon and imatinib (IRIS), confirmed the efficacy and safety of imatinib in the long term with an overall survival of 85% and an event-free survival of 81%. However, some mutations in the kinase domain BCR-ABL confer resistance to one or more TKI, influencing the choice of therapy, as in the case of a T315I mutation, which is highly resistant to imatinib. Although many factors contribute to the resistance to imatinib, the presence of mutations is more prevalent and has been further investigated. Therefore, were aimed to perform mutation analysis in patients with a resistant phenotype of two cancer reference hospitals in Vitoria, ES, Brazil. We also developed a CML database relating clinical information and laboratory cytogenetics and molecular biology results, facilitating the monitoring of CML patients, as well as the understanding of disease progression.
Efeitos quimioprotetivos do flavonoide hesperidina contra mutagenicidade induzida por cisplatina em medula óssea de camundongos
(Universidade Federal do Espírito Santo, 2013-03-25) Passos, Tatiane da Silva; Batitucci, Maria do Carmo Pimentel; Matsumoto, Silvia Tamie; Paula, Flavia de
Hesperidin is a flavonoid naturally found in citrus fruit especially orange and lemon peel. We investigated the antimutagenic activity of this compound against the cytotoxic and mutagenic effects induced by the anticancer drug cisplatin (CddP) using the micronucleus assay in bone marrow erythrocytes of mice (Mus musculus). We used three protocols: pre-treatment with hesperidin (by gavage) for 14 days, followed by applying (intraperitoneally) from CddP no 14th day; Post-treatment administration of CDDP and 24 hours after the administration of hesperidin and simultaneous treatment with hesperidin and CddP. We evaluated three concentrations of Hesperidin diluted in corn oil (100mg.kg-1, 200mg.kg-1 e 400mg.kg- 1). The controllling was conducted by the negative control (NaCl 0,9%), positive control (CddP 0,05mg.kg-1) and and the solvent controls (corn oil). For micronucleus test, the animals were euthanized 24 hours following the last treatment by cervical dislocation, and bone marrow was collected. The samples were transferred to Falcon tubes and centrifuged. Slides were prepared by smear technique with a drop of suspension and fixed with methanol for 10 minutes, staining was performed with Leishman stain. The reading was performed in blind test, we analyzed 2000 cells per animal in an optical microscope for evaluation of micronucleated polychromatic erythrocytes (MNPCEs) and the ratio PCE/(PCE+NCE) (polychromatic erythrocytes/polychromatic erytrocites + normochromatic erythrocytes). All protocols studied showed a reduction in the number of micronuclei when compared to CP and achieved high rates of reduction of damage caused by cisplatin, except the concentration of 400 mg.kg-1 of the simultaneous treatment protocol where such protective effect was not observed indicating that hesperidin has chemoprotective action on the DNA molecule. It is known that various flavonoids have the ability to eliminate free radicals such as hydroxyl radicals generated by chemical agents that cause DNA damage, but the exact mechanism of how hesperidin acts is not yet clear.
Efeito da alta pressão hidrostática em sementes de Carica papaya
(Universidade Federal do Espírito Santo, 2013-04-18) Santos, Alan Costa Sarcineli; Fernandes, Patricia Machado Bueno; Fernandes, Antonio Alberto Ribeiro; Costa, Aureliano Nogueira da; Silva, Diolina Moura
The Carica papaya originates from tropical America. Brazil is among the world's major producers, and the state of Espírito Santo figure as the country's biggest exporter, the variety of the group is more cultivated soil, and crops for export, a variety prevails Golden Land Group (Jacomino , BROWN, KLUGE, 2003). The state produces more than 630,000 tons annually, approximately, 7,900 there. The cultivation generates a gross income of approximately $ 50 million Porano and employs about 9,000 people in the production and marketing throughout the year, which makes this fruit, among the most important of the exploited in this state.

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