Navegando por Palavras-chave "Mouse"

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    Alterações degenerativas induzidas pela privação de sono paradoxal em glândula sublingual de ratos
    (Universidade Federal de São Paulo, 2023-04-26) Aguiar, Gabriel Carvalhal de [UNIFESP]; Ribeiro, Daniel Araki [UNIFESP]; Souza, Ana Carolina Flygare [UNIFESP]; http://lattes.cnpq.br/3721735477545941; http://lattes.cnpq.br/9969803499258672; http://lattes.cnpq.br/4018439041598806; Universidade Federal de São Paulo (UNIFESP)
    O ser humano passa cerca de um terço de sua vida dormindo, processo este fundamental para a manutenção e equilíbrio dos mecanismos psicossocial e biológico. O sono é um processo natural e complexo, e a falta dele pode acarretar e/ou agravar problemas de saúde, tais como diabetes, hipertensão, depressão, diminuição da função imunológica, dentre outros. Nesse contexto, o presente estudo teve como objetivo avaliar se a privação de sono é capaz de deflagrar processo proliferativo e apoptótico nas glândulas sublinguais de ratos. Para isso, 24 ratos foram distribuídos em três grupos, Controle (n=8), no qual os animais não foram submetidos a qualquer procedimento; Privação do Sono (n=8) os animais foram submetidos por um período de 96 horas consecutivas à privação de sono; e Rebote (n=8), os animais foram submetidos à privação de sono paradoxal por 96 horas consecutivas, seguidos por 96 horas sem intervenção. Foram investigadas alterações histopatológicas e imunoexpressões das proteínas Ki-67, p16, Caspase-3 clivada e BCL-2. Os resultados demonstraram que a privação de sono paradoxal induziu degeneração tecidual a partir da presença de picnose, vacúolos e áreas de retenção salivar, nos grupos experimentais. A expressão de Caspase 3 clivada e bcl2 aumentou tantos nos grupos privação de sono como rebote. A análise do ki-67 demonstrou um aumento da expressão somente no grupo rebote, associado à diminuição da proteína p16. Com isso, a privação do sono se mostra capaz de deflagrar processo degenerativo no parênquima glandular, por meio da desregulação da apoptose e atividade proliferativa na glândula sublingual de ratos.
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    Amifostine Increases FAS and Caspase-3 Expression in Colonic Tissue of Irradiated Mice
    (Int Inst Anticancer Research, 2015-05-01) Oshima, Celina Tizuko Fujiyama [UNIFESP]; Ribeiro, Daniel Araki [UNIFESP]; Gomes, Thiago Simao [UNIFESP]; Adios, Priscila C. [UNIFESP]; Egami, Mizue Imoto [UNIFESP]; Segreto, Helena Regina Comodo [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Background/Aim: The aim of this study was to evaluate the expression of FASL, FAS and FADD and caspase-3 in oesophagus, stomach and colonic tissues of mice irradiated in vivo by immunohistochemistry. Materials and Methods: A total of 48 adult male C57BL mice were distributed into four groups: Ami(-)/Rad(-): Mice received 0.5 ml of 0.9% physiological saline solution (PPS) intraperitioneally (i.p.); Ami(+)/Rad(-): mice received amifostine (400mg/kg i.p.) freshly dissolved in double-distilled water; Ami(-)/Rad(+): mice received 0.5 ml of PSS i.p. 30 min before a single whole-body radiation dose of 7 Gy; Ami(+)/Rad(+): mice received 0.5 ml of an aqueous solution of 400 mg/kg amifostine i.p.30 min prior to irradiation. All groups were assigned into subgroups sacrificed at 0.5 h, 1 h, 2 h and 4 h after irradiation. Results. In oesophagus and stomach tissues, we did not observe any difference between Ami(-)/ad(-), Ami(+)/Rad(-), Ami(-)/Rad(+) and Ami(+)/Rad(+) groups in the expression of FASL, FAS and FADD. The colonic tissue was the only to exhibit any difference in the expression of FAS and caspase-3 protein in the Ami(-)/Rad(+) group at 1 and 2 h. Amifostine increased FAS and caspase-3 immunoexpression when compared to the control. Immunoexpression for FASL and FADD was not remarkably different in colonic tissue. Conclusion: Taken together, our results demonstrate that amifostine increases FAS and caspase-3 expression in colonic tissue of irradiated mice.
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    Concentration of glycosaminoglycan in ovariectomized mice uterus after treatment with ovarian steroids
    (Hindawi Publishing Corp, 2016) Maioral, Gabriela C. C. C. [UNIFESP]; Gomes, Regina Celia T. [UNIFESP]; Verna, Carina [UNIFESP]; Simoes, Manuel de J. [UNIFESP]; Nader, Helena B. [UNIFESP]; Simoes, Ricardo S.; Baracat, Edmund C.; Soares-, Jose Maria, Jr.
    The aim of this study was to evaluate the amount of non-and sulfated glycosaminoglycans in the ovariectomized mice uterus, after treatment with ovarian steroids. For this purpose, 50 adult female mice were divided into five groups with 10 animals/each: control group: CG (ovary intact), and ovariectomized groups: OG (vehicle), EG (estradiol), PG (progesterone) and EPG (estradiol combined to progesterone). The treatments started 30 days after ovariectomy. All the animals were treated for 50 consecutive days. These hormones were administered in a sterile oily solution via gavage. Twenty-four hours after the last treatment, all animals were euthanized, removing the uterine horn for biochemical analyses. To quantify, the hyaluronic acid (HA) used ELISA-like fluorometric assay, and the sulfated glycosaminoglycans (GAGs) used agarose gel electrophoresis. The amount of HA was significantly higher in the group treated with progesterone (PG) compared to the others groups (p < 0.05), and in the group treated with estradiol (EG), the amount of chondroitin/dermatan sulfate was significantly higher compared to the others groups (p < 0.05), and in the group treated with progesterone (PG), the amount of heparan sulfate was significantly lower compared to the others groups, except to control group (p < 0.05). Our results showed that the estroprogestative therapy after long time (50 days) profoundly affected the amount of glycosaminoglycans in uterine. These changes may be indicative of uterine pathology such as the development of tumor.
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    Efeito da adição do LH durante o estímulo ovariano e maturação oocitária no perfil lipídico de oócitos murinos
    (Universidade Federal de São Paulo (UNIFESP), 2016-02-02) Oleinki, Talitha Dinardo [UNIFESP]; Fraietta, Renato [UNIFESP]; http://lattes.cnpq.br/1545035937368744; http://lattes.cnpq.br/7699135183138091; Universidade Federal de São Paulo (UNIFESP)
    Introduction: The addition of LH in assisted reproduction aiming to obtain a higher number of viable follicles for the treatment is still controversial. A good embryonic development depends on the follicular growth and proper oocyte maturation. Because of that, the association between alkaline comet assay and the study of oocyte lipidomics may help to understand the biological processes involved in oocyte maturation and the influence of hormonal protocol. This study may contribute in the future with effective hormonal protocols, improving the quality of oocytes and pregnancy rates. Objective: Evaluate the effect of adding LH on DNA integrity of cumulus cells and on lipid profile of murine oocytes. Methods: Female mice C57BL / 6J 23 to 28 days old received three different stimulation protocols intraperitoneally (i.p.): (i) only PMSG; (ii) PMSG plus hCG (iii) PMSG plus LHr. After 48 hours of administration, immature oocytes were collected and cultured for 24 hours in three different culture medium for maturation: (i) only FSH; (ii) FSH plus hCG (iii) FSH plus LHr. After this period, the oocytes which had the first polar body were frozen at -80oC until the analysis by electrospray ionization (ESI) and cumulus cells were subjected to alkaline comet technique to measure the integrity of DNA. The principal component analysis and discriminant analysis by least squares were performed and 35 ions with greater representation were identified by the variable importance in the projection. Results: There was no statistical difference in the maturation rate and DNA fragmentation of cumulus cells in different groups analyzed. The fingerprinting analysis of the lipid profile of oocytes matured in vitro identified twenty lipids. The hyper-represented lipids in the group stimulated with the addition of hCG and matured in culture medium only with FSH (HF group) are: phosphatidylethanolamine, polyketide (flavonoid), sterol, phosphatidylserine, polyketide (ansamycin), sphingomyelin and phosphatidylcholine; in the group stimulated and matured in the presence of hCG (HH group) phosphatidylglycerol, phosphatidylserine, sphingolipids and triacylglycerol are hyper-represented; and in group stimulated with addition of LHr and matured in culture medium only with FSH (LF group) the polyketide (flavonoid) and sphingolipids are hyper-represented. Conclusion: The protocol used for hormonal stimulation and MIV modify the lipid profile of the oocyte, but does not alter the oocyte maturation rate and the DNA integrity of cumulus cells.
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    Efeito da nicotina sobre o epitélio seminífero de ratos expostos durante a fase intrauterina e a lactação
    (Universidade Federal de São Paulo (UNIFESP), 2015-05-27) Paccola, Camila Cicconi [UNIFESP]; Valdeolivas, Sandra Maria Miraglia [UNIFESP]; http://lattes.cnpq.br/7489168456011888; http://lattes.cnpq.br/8116023354222658; Universidade Federal de São Paulo (UNIFESP)
    A nicotina é uma droga lícita de alto consumo mundial, por meio do cigarro. Ela é capaz de atravessar a membrana placentária e está presente no leite de mães tabagistas. Esta droga induz apoptose em diversos órgãos, altera a secreção de hormônios sexuais e provoca infertilidade masculina. Para investigar se a exposição à nicotina durante a gestação e a lactação induz apoptose nas células do epitélio seminífero ou se altera a morfologia e a função das células de Sertoli comprometendo secundariamente as células da linhagem germinativa, ratas prenhes receberam nicotina (2mg/Kg/dia) através de uma mini-bomba osmótica. As proles masculinas (30, 60 e 90 dias pós-natais) tiveram o sangue coletado para dosagem hormonal (FSH e LH) e os testículos submetidos à análise histopatológica, com análises das frequências dos estágios do ciclo do epitélio seminífero. A ocorrência de apoptose (pelo método TUNEL e imuno-marcação das proteínas Fas e FasL), a função e estrutura de células de Sertoli (respectivamente, pela imuno-marcação de transferrina e vimentina), e a molécula juncional entre células de Sertoli e células germinativas (pela imuno-marcação da ?-catenina) foram avaliadas. A exposição à nicotina provocou aumento dos níveis plasmáticos de FSH e LH nos animais adultos. Embora a exposição à nicotina não tenha alterado a quantidade de células apoptóticas e nem a expressão das proteínas Fas e FasL, ela provocou uma intensa descamação de células epiteliais testiculares e alterou a frequência de alguns estágios do ciclo do epitélio seminífero na puberdade tardia e fase adulta. As expressões de transferrina e ?-catenina não foram alteradas, mas a vimentina foi significantemente reduzida nos estágios iniciais do ciclo do epitélio seminífero dos animais adultos expostos à nicotina. Assim, concluiu-se que a exposição à nicotina durante a gestação e a lactação, altera a expressão da vimentina, uma proteína importante do citoesqueleto das células de Sertoli e este evento que pode ter sido decorrente da alteração dos níveis de gonadotrofinas hipofisárias como também o responsável pela intensa descamação de células da linhagem germinativas dos animais na fase adulta. Portanto, esta pesquisa encoraja o abando do cigarro pela gestante fumante, diante da possibilidade de comprometimento da espermatogênese da prole, além de todos os malefícios já bem conhecidos relacionados ao consumo de cigarros.
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    Envenenamento experimental por Bothrops jararaca em camundongos prenhes: Eficácia do soro antibotrópico
    (Universidade Federal de São Paulo (UNIFESP), 2011-03-30) Ferreira, Karla Vanessa [UNIFESP]; Godosevicius, Sima [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Introduction: Snakebite accidents are considered a rare event among pregnant women, but serotherapy is indicated even when envenomation is not severe. However, antivenom can cause maternal adverse reactions and consequently fetal death. Experimental Bothrops jararaca (Bj) envenomation can provoke marked morphological alterations in the antimesometrial (AM) region of uterus in pregnant mice which can culminate in the end of gestation. This investigation aimed to verify whether Bothrops antivenom (BAV) could restore the normal morphology of murine uterus, after Bj envenomation. Methods: On the morning of day 7,5 of pregnancy, animals received Bj venom (0.24mg Bj venom/kg body weight) i.m., and after 3 hours they were treated with BAV i.v. (VBj+BAV). Control groups received saline and was treated with BAV (Sal+BAV) or Bj venom (VBj). On day 8,5, uterine morphology was analyzed, especially at the maternal-fetal interface in the AM region. Plasma fibrinogen (Fg) was assayed in plasma samples of pregnant animals. Aiming to study the external appearance and the skeletal morphology of fetuses, as well as the incidence of fetal resorptions, another group of animals on day 7,5 received the same treatments mentioned and was sacrificed on day 18,5. Results: Histological analysis of most dams of the VBj group revealed the maternal and fetal tissues disorganized, showing hemorrhagic areas and a prominent inflammatory infiltrate at the maternal-fetal interface. Additionally, decidual cells (maternal) and trophoblastic giant cells (fetal) exhibited evident signs of necrosis. The antimesometrial decidua of most dams of the VBj+BAV revealed the maternal fetal tissues organized, similarly to the uteri of dams of the Sal+BAV group. Analyses of the external appearance and skeletal morphology of fetuses on day 18,5 showed no difference among groups; however, the dams that received Sal+BAV and were treated with ABS showed smaller fetuses than VBj+BAV group. Plasma Fg levels of the VBj+BAV group were similar to those of Sal+BAV group. However, experimental Bj envenomation showed lower plasma Fg levels. Conclusion: Our results indicated that the treatment with the antibothropic serum was capable to maintain the development of the gestation, preserving the morphology in the maternal-fetal interface, in the animals.
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    Homocysteine and cysteine concentrations are modified by recent exposure to environmental air pollution in São Paulo, Brazil
    (Elsevier B.V., 2009-10-01) Abe, Karina Camasmie [UNIFESP]; Brandao, Leticia de Campos [UNIFESP]; Aguilar Calegare, Bruno Frederico [UNIFESP]; Tufik, Sergio [UNIFESP]; Nascimento Saldiva, Paulo Hilario do; D'Almeida, Vania [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)
    Millions of people worldwide are affected by anthropogenic air pollution derived from the combustion of fossil fuels. in this work, we tested the effects of fetal, lactation and post-weaning ambient air pollution exposure on total homocysteine (tHcy) concentrations and on a downstream pathway element, the plasma cysteine (Cys) concentration. Two similar exposure chambers (polluted and filtered chamber) were located near an area with heavy traffic in São Paulo, Brazil, and male Swiss mice were housed there from the pre-natal period until 3 months of age. Groups during fetal, lactation and adult periods of exposure were apportioned, and tHcy and Cys plasma concentrations were assessed when the animals were 3 months old. in our study, both the tHcy and Cys concentrations were decreased in groups that spent their final stage of life in polluted chambers, suggesting recent alterations in tHcy and Cys concentrations due to air pollution exposure. the possible relationship of these data with cardiovascular dysfunction is still a matter of controversy in animals; nevertheless, epigenetic mechanisms emerge as a possible issue to consider in the investigation of the link between air pollution and Hcy measurement. (C) 2009 Elsevier Inc. All rights reserved.
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    Inducing mutations in the mouse genome with the chemical mutagen ethylnitrosourea
    (Associação Brasileira de Divulgação Científica, 2006-09-01) Massironi, Sílvia Maria Gomes; Reis, B.l.f.s.; Carneiro, Juliana G.; Barbosa, Luciene B. S.; Ariza, Carolina Batista [UNIFESP]; Santos, Gilmara Cristina dos; Guénet, Jean Louis; Godard, Ana Lúcia Brunialti; Universidade de São Paulo (USP); Universidade Federal de Minas Gerais Instituto de Ciências Biológicas Departamento de Biologia Geral; Universidade Federal de São Paulo (UNIFESP); Institut Pasteur Département de Biologie du Développement
    When compared to other model organisms whose genome is sequenced, the number of mutations identified in the mouse appears extremely reduced and this situation seriously hampers our understanding of mammalian gene function(s). Another important consequence of this shortage is that a majority of human genetic diseases still await an animal model. To improve the situation, two strategies are currently used: the first makes use of embryonic stem cells, in which one can induce knockout mutations almost at will; the second consists of a genome-wide random chemical mutagenesis, followed by screening for mutant phenotypes and subsequent identification of the genetic alteration(s). Several projects are now in progress making use of one or the other of these strategies. Here, we report an original effort where we mutagenized BALB/c males, with the mutagen ethylnitrosourea. Offspring of these males were screened for dominant mutations and a three-generation breeding protocol was set to recover recessive mutations. Eleven mutations were identified (one dominant and ten recessives). Three of these mutations are new alleles (Otop1mlh, Foxn1sepe and probably rodador) at loci where mutations have already been reported, while 4 are new and original alleles (carc, eqlb, frqz, and Sacc). This result indicates that the mouse genome, as expected, is far from being saturated with mutations. More mutations would certainly be discovered using more sophisticated phenotyping protocols. Seven of the 11 new mutant alleles induced in our experiment have been localized on the genetic map as a first step towards positional cloning.
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    Retinal and Ocular Toxicity in Ocular Application of Drugs and Chemicals - Part I: Animal Models and Toxicity Assays
    (Karger, 2010-01-01) Penha, Fernando Marcondes [UNIFESP]; Rodrigues, Eduardo B. [UNIFESP]; Maia, Mauricio [UNIFESP]; Dib, Eduardo [UNIFESP]; Costa, Elaine Fiod [UNIFESP]; Furlani, Bruno A. [UNIFESP]; Moraes Filho, Milton Nunes [UNIFESP]; Dreyfuss, Juliana L. [UNIFESP]; Bottos, Juliana [UNIFESP]; Farah, Michel E. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP)
    Aims: Experimental retinal research has gained great importance due to the ophthalmic pharmacotherapy era. An increasing number of drugs are constantly released into the market for the treatment of retinal diseases. in this review, animal species, animal models and toxicity assays in retinal research are discussed. Methods: An extensive search of the literature was performed to review various aspects of the methods of investigation of drug toxicity. the different types of animal species, as well as single animal models available for the evaluation of safety and efficacy of retinal pharmacotherapy, were identified. in addition, a large variety of reported laboratory techniques were critically examined. Results: in vitro studies are the first-line experiments for the development of a new drug for retinal diseases, using retinal pigment epithelial cells and other cell lines. the next step involves in vivo animal studies where nonhuman primates are considered the gold standard. However, cost and legal issues make their use difficult. Mice and rats provide genetically controlled models for investigations. Pigs, dogs and cats represent good large-size animal models, while rabbits are one of the most used species for retinal toxicity evaluations. Various laboratory methods were identified, including light microscopy, electron microscopy, electroretinography and new emerging methods, such as optical coherence tomography and scanning laser ophthalmoscopy for experimental purposes. Conclusions: A great number of animal species and models are available that simulate retinal diseases and provide experimental data for further human use. Work with animal models should include properly designed toxicity assays to obtain reliable results for safety and efficacy. Copyright (C) 2010 S. Karger AG, Basel
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    Simplified estimates of ion-activity products of calcium oxalate and calcium phosphate in mouse urine
    (Springer, 2012-08-01) Tiselius, Hans-Goran; Nogueira Ferraz, Renato Ribeiro [UNIFESP]; Heilberg, Ita Pfeferman [UNIFESP]; Karolinska Inst; Universidade Federal de São Paulo (UNIFESP)
    This study aimed at formulating simplified estimates of ion-activity products of calcium oxalate (AP(CaOx)) and calcium phosphate (AP(CaP)) in mouse urineto find the most important determinants in order to limit the analytical work-up. Literature data on mouse urine composition was used to determine the relative effect of each urine variable on the two ion-activity products. AP(CaOx) and AP(CaP) were calculated by iterative approximation with the EQUIL2 computerized program. the most important determinants for AP(CaOx) were calcium, oxalate and citrate and for AP(CaP) calcium, phosphate, citrate, magnesium and pH. Urine concentrations of the variables were used. A simplified estimate of AP(CaOx) (AP(CaOx)-index(MOUSE)) that numerically approximately corresponded to 10(8) x AP(CaOx) was given the following expression:AP(CaOx)-index(MOUSE) = 0.70 x Calcium(1.05) x Oxalate(0.95)(0.90-0.0225 x Citrate) + (6.6 x 10(-8) x Citrate(3.98))For a series of urine samples with various composition the coefficient of correlation between AP(CaOx)-index(MOUSE) and 10(8) x AP(CaOx) was 0.99 (p = 0.00000). A similar estimate of AP(CaP) (AP(CaP)-index(MOUSE)) was formulated so that it approximately would correspond numerically to 10(14) x AP(CaP) taking the following form:Ap(CaP)-index(MOUSE) = 0.05 x Calcium(1.17) x Phosphate(0.85) x Magnesium(0.18) x (pH - 4.5)(6.8) / Citrate(0.76)For a series of variations in urine composition the coefficient of correlation was 0.95 (p = 0.00000). the two approximate estimates shown in this article are simplified expressions of AP(CaOx) and AP(CaP). the intention of these theoretical calculations was not to get methods for accurate information on the saturation levels in urine, but to have mathematical tools useful for rough conclusions on the outcome of different experimental situations in mice. It needs to be emphasized that the accuracy will be negatively influenced if urine variables not included in the formulas differ very much from basic concentrations.