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
Indução de resistência ao Papaya meleira virus em mamoeiros
(Universidade Federal do Espírito Santo, 2011-03-04) Abreu, Paolla Mendes do Vale de; Ventura, José Aires; Fernandes, Patricia Machado Bueno; Loureiro, Marcelo Ehlers; Louro, Iúri Drumond
Carica papaya L. is one of the most cultivated and consumed fruits in tropical and subtropical regions of the world. Brazil is the world's largest producer of papaya and the states of Espirito Santo and Bahia are responsible for more than 70% of the Brazilian production of this fruit. Diseases of fruit trees are the main limiting factors. Papaya meleira disease, caused by the Papaya meleira virus (PMeV), is one disease that does not have a resistant cultivar. However, the PMeV genome is a doublestranded RNA (dsRNA) and it has been shown that virus resistance in plants can be activated by dsRNA molecules. After detecting the presence of dsRNA, the cell initiates a process of degradation of RNA molecules, which may be viral, thus preventing the progress of the infection. This study aimed to induce resistance to PMeV in papaya seedlings using dsRNA molecules extracted from the viral genome. Four different treatments were evaluated qualitatively and quantitatively by means of molecular diagnosis of the virus in leaf samples by RT-PCR and qRT-PCR (real-time PCR), respectively. The papaya seedlings inoculated only with high load PMeV showed severe infection in the first days post-inoculation, while the seedlings inoculated with high viral load and additional dsRNA showed a milder viral infection, suggesting a reduction in infection by the virus that causes meleira. The combined inoculation of papaya seedlings with PMeV and viral dsRNA reduced the progress of the infection.
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.
Detecção molecular do complexo viral da murcha do abacaxizeiro em plantas e no inseto vetor
(Universidade Federal do Espírito Santo, 2011-05-03) Amorim, Walkíria Andrade de; Fernandes, Patricia Machado Bueno; Ventura, José Aires; Costa, Hélcio; Culik, Mark Paul
Brazil is one of the largest producers of pineapple, but the crop can have high economic losses due to diseases like mealybug wilt, caused by one or more Pineapple mealybug wilt-associated viruses (PMWaVs) with yield losses up to 80%. This study aimed to develop a methodology to detect the virus complex in plants and insect vectors, efficiently, simply and at low cost. Samples of plants and mealybugs were collected at the Experimental Farm of Incaper in Sooretama, at the Centro Serrano Regional Development Center in Domingos Martins, and at commercial plantations in the municipality of Marataízes, Espírito Santo, for virus detection by RT-PCR using degenerate primers. The results showed that the samples must be processed in the laboratory within 48 hours of collection or stored in a freezer for a maximum of 30 days. For samples of plants in vitro, and the green, pigmented leaf region and roots of plants from the field, the best protocol for RNA extraction was found to be Trizol®, and for chlorotic, unpigmented leaf tissues the method of Doyle and Doyle (1990) was preferable. In older leaves of plants from the field, we could not detect the virus. It was observed that absence of wilt symptoms in plants did not guarantee absence of the virus. In analysis of plants in tissue culture, we detected the presence of the virus in 15% of samples. The technique for viral diagnosis in mealybugs was standardized using degenerate and specific primers and Trizol® reagent, from the protocol of Gibbs and Mackenzie (1997). Although the mealybug Dysmicoccus brevipes is considered a vector, this is the first time that PMWaV has been detected in this insect by PCR with three PMWaVs detected in the same insect. The new methodology developed in this research was shown to be viable for the diagnosis of disease, disease indexing of propagative material, and detection of the virus in insect vector.
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.
Triagem de mutações no gene COL1A1 em pacientes com osteogênese imperfeita
(Universidade Federal do Espírito Santo, 2011-08-26) Moraes, Marcos Vinícius Dornelas de; Paula, Flávia de; Errera, Flávia Imbroisi Valle; Bortolini, Eliete Rabbi
Osteogenesis Imperfecta (OI) is a heritable disorder associated with bone fragility and propensity to fracture. The most common mutations associated with dominant inheritance affect the structural genes of type I collagen and can result from failure to synthesize the products of COL1A1 or COL1A2 genes (quantitative) or by substitution for glycine residues within the Gly-X-Y triplet domain of the triple helix (qualitative). The aim of this study was to verify the occurrence of pathogenic mutations in coding regions of the COL1A1 gene, including splice sites and exon flanking regions, in patients with Osteogenesis Imperfecta assisted at the Hospital Infantil Nossa Senhora da Glória, Vitória - ES.
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.
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.
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
Avaliação de danos genômicos em pacientes submetidos a hemodiálise, após suplementação vitamínica
(Universidade Federal do Espírito Santo, 2012-03-02) Batista, Thatiane Lorena Miranda; Batitucci, Maria do Carmo Pimentel; Meyrelles, Silvana dos Santos; Bortolini, Eliete Rabi
Chronic renal failure (CRF) is increasingly common in the population and involves the irreversible loss of kidney function necessitating treatment of renal replacement therapy. The most common is hemodialysis that performs a bypass filtration, removing fluids and toxins from the blood of the patient. But this process does not physiological causes an increase in oxidative stress and complications such as diseases of morbidity and mortality among them the cardiovascular disease and cancer. For a reduction in the frequency of these diseases is achieved the use of vitamins is necessary because the nutritional status affects the development of these diseases. The aim of this study was to examine the genotoxicity and cytotoxicity in chronic renal patients using the micronucleus assay in lymphocytes and buccal mucosa cells after vitamin supplementation by folic acid and vitamins E, C and B12. There were 16 voluntary patients provided that each of the three samples for each test, a sample was received before supplementation and during the treatment, after one and two months of use of vitamins. In the assay of lymphocytes with blocking cytokinesis (CBMN) were analyzed nuclear buds, nucleoplasmic bridges and micronuclei in binucleated cells, and nuclear division index. Already in the cells of mucosa were binucleated cells, mononucleated and apoptosis, karyolysis, micronucleus and broken egg. The results show that the frequency of nuclear abnormalities was reduced with the use of vitamins in two trials, and in some assessments there were no statistical differences. In addition, were found gender differences in behavior of supplementation. This study provides evidence that vitamin use by patients with chronic renal failure was associated with reductions in nuclear changes evaluated, which may contribute to better quality of life for these patients.
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.
Avaliação in vitro da atividade antimicrobiana e antioxidante de extratos fitoterápicos produzidos na Pastoral da Saúde de Venda Nova do Imigrante-ES
(Universidade Federal do Espírito Santo, 2012-05-31) Baptista, Lilliane Bonella Meireles; Nunes, Ana Paula Ferreira; Batitucci, Maria do Carmo Pimentel; França, Hildegardo Seibert
The medicinal properties of herbs used traditionally by the population have been proved by researches from all around the world. Antimicrobial and antioxidant activities of eight phytotherapeutic extracts produced in the Pastoral care of health of Venda Nova do Imigrante-ES were assessed, from the plants: Anadenanthera colubrina (Vell.) Brenan, Achillea millefolium L., Aristolochia cymbifera Mart., Casearia sylvestris Sw., Cordia verbenacea DC., Echinodorus grandiflorus (Cham.& Schltdl.) Micheli, Gossypium hirsutum L. and Plantago major L. The antimicrobial activity was evaluated by the microdiluition method for determining the minimal bactericidal concentration (MBC) of each extract against 12 bacterial species, including multi-drug resistant strains causing nosocomial infections like MRSA, VRE and ESBL K.pneumonie. All plant extracts, with the exception of A. cymbifera whose MBC > 250 mg/mL, showed antimicrobial activity against the tested bacteria, with MBC ranging between 4 and 86 mg/mL to Gram-positive bacteria and Gram-negative bacteria, with special highlights for the extracts of A. colubrina and C. verbenacea, for which there was the kinetic evaluation test of bacterial death (time-kill). The antioxidant activity of extracts was evaluated by the DPPH method, demonstrating that the extracts are herbal medicines of regular to great antioxidant action, compared to standard quercetin. The results obtained confirm empirical indications attributed to the phytotherapeutic extracts including antimicrobial action, corroborating to assign new uses for the phytotherapics evaluated, as well as to encourage further studies with the extracts and the plants from which they are derived
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.

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