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Embryonic stem-like cells derived from in vitro produced bovine blastocysts BABT
Freitas,Erika Regina Leal de; Sanches,Bruno Valente; Gambarini,Maria Lúcia; Oliveira Filho,Benedito Dias; Guillo,Lidia Andreu.
The aim of this work was to study the derivation of bovine embryonic stem-like (ES-like) cells from the inner cell mass (ICM) of in vitro produced blastocysts. The ICMs were mechanically isolated and six out of seventeen (35%) ICMs could attach to a monolayer of murine embryonic fibroblasts (MEF). Ten days after, primary outgrowths were mechanically dissected into several small clumps and transferred to a new MEF layer. Cells were further propagated and passaged by physical dissociation over a 60 days period. The pluripotency of the bovine ES-like cells was confirmed by RT-PCR of Oct-4 and STAT-3 gene markers. The colonies were weakly stained for alkaline phosphatase and the mesoderm and endoderm differentiation gene markers such as GATA-4 and Flk-1,...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Bovine blastocysts; ICM isolation; Embryonic stem cells; Gene markers.
Ano: 2011 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132011000300009
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Produção de Lactobacillus plantarum em Melaço de Cana-de-açúcar BABT
Villavicencio,Angel R. N.; Sant’Anna,Ernani S.; Tôrres,Regina C. O..
Comparative growth studies of Pediococcus pentosaceus were done in MRS (control) culture medium as well as in culture medium of 5% sugar cane blackstrap molasses (Broth 1) enriched with yeast extract (Broth 2) and enriched with beef extract (Broth 30. The experiment was carried out in a shaker (96 rpm) in 250 mL Erlenmeyers flasks with working volume of 150 mL, initial inoculum of about 10³ - 10(4) FCU/mL, at 35 ± 1ºC temperature and 28-hour fermentation. The biomass pruductivity was higher in MRS broth (0.0375 g/L.h), followed by Broth 2 (0.0275), Broth 3 (0.0189) and Broth 1 (0.0151). The higher viable cells number occured in MRS (13.96 log10 FCU/mL), followed by Broth 2 (12.09), Broth 3 (10.12) and Broth 1 (8.42).
Tipo: Info:eu-repo/semantics/article Palavras-chave: Pediococcus pentosaceus; Starter culture; Molasses.
Ano: 1999 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89131999000200015
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Role of Natural Antioxidants in the Modulation of Plasma Amino Acid Pattern in Rats Exposed to Hemic Hypoxia BABT
Al-Rasheed,Nouf Mohamed; Fadda,Laila Mohamed; Al-Rasheed,Nawal Mohamed; Attia,Hala; Ali,Hanaa Mahmoud; El-Agami,Hanaa.
ABSTRACTThe aim of this work was to investigate whether the free radical scavengers, L-arginine (L-arg) and/or carnosine, either alone, or in combination would modulate tissue injury induced by hypoxia by measuring Fischer's ratio [concentrations of branched chain amino acids (BCAAs)/aromatic amino acids]. Decreased Fischer's ratios and increased malondialdehyde (MDA) led to pathogeneses of many diseases. Rats were injected with sodium nitrite (60 mg/kg) to establish hypoxia. They were treated with L-arg, (200 mg/ kg) and/or carnosine (200 mg/ kg) and their combination 24 and 1 h prior to sodium nitrite intoxication. The results revealed that hypoxia significantly decreased hemoglobin, arginine, citrulline and proline and increased sLDH, MDA , ammonia ,...
Palavras-chave: Natural antioxidants; Amino acids; Carnosine; L-arginine; Fisher's ratio; Urea.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000500741
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Gills of juvenile fish piaractus mesopotamicus as histological biomarkers for experimental sub-lethal contamination with the Organophosphorus Azodrin®400 BABT
Rudnicki,Cácia Aparecida Mendes; Melo,Gisele Cristiane; Donatti,Lucélia; Kawall,Helena Gonçalves; Fanta,Edith.
Piaractus mesopotamicus is a freshwater native fish from rivers of the Paraná-Paraguay Basin and of the Pantanal region and has been used for repopulation programs in Brazil. Juvenile fishes were exposed to the sub-lethal dose of 1.08mg/L of the OP Azodrin®400 containing 0.43µL/L of the active principle monocrotophos for 96 h. A frequent pathology in the gills at all times of exposure was epithelial detachment, from minimal until 24 h of exposure, to intense after 48 h of contamination. Deformed pillar cells in the respiratory lamellae leading to irregular blood spaces and blood congestion, as well as hyperplasia and lamellar fusion were observed. These histopathologies suggested that 48 h after T0 was an important time when a reduction in the capability...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Freshwater Fish; Organophosphorus; Monocrotophos; Morphology; Branchiae.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000600015
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Composition and significance of glycosaminoglycans in the uterus and placenta of mammals BABT
Oliveira,Gleidson Benevides de; Vale,André Menezes do; Santos,Amílton Cesar dos; Moura,Carlos Eduardo Bezerra de; Rocha,Hugo Alexandre de Oliveira; Oliveira,Moacir Franco de.
Glycosaminoglycans (GAGs) are heteropolysaccharides in mammalian tissue and consist of repeated disaccharide units with mono-sulfated or non-sulfated monosaccharides. GAGs are important components of the Extracellular Matrix (ECM) with several physiological roles, in the recognition, migration, adhesion, proliferation and differentiation processes. They are also important in angiogenesis, blood homeostasis, immune reactions, follicule development and also in the development of pathologies such as infertility, tumors and metastases. It has been shown that the profile of glycosaminoglycans in the uterine and placental tissues is highly variable throughout the reproductive cycle and during pregnancy. It may be directly related to their physiological or...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Extracellular matrix; Hormonal fluctuation; Reproductive events; Euterium mammals; Pathology.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000400512
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Combination vitamin C and vitamin E prevents enteric diabetic neuropathy in the small intestine in rats BABT
Zanoni,Jacqueline Nelisis; Hermes-Uliana,Catchia.
The present study evaluated the effects of supplementation with a combination of vitamin C and vitamin E on NADH-diaphorase-positive (NADH-d+) and neuronal nitric oxide synthase (nNOS)-immunoreactive myenteric neurons in the duodenum and ileum in diabetic rats. Forty rats were distributed into the following groups: normoglycemic (N), normoglycemic supplemented with vitamin C and vitamin E (NS), diabetic (D), and diabetic supplemented with vitamin C and vitamin E (DS). Vitamin C was added to the drinking water, and vitamin E was incorporated in the diet (1%). After 120 days, the animals were euthanized, and the duodenum and ileum were subjected to NADH-d and nNOS staining. Quantitative and morphometric analyses of myenteric neurons were performed. Diabetes...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Ascorbic acid; Α-tocopherol; Diabetes mellitus; Myenteric plexus; Enteric nervous system.
Ano: 2015 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132015000400504
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Nanofibrous Scaffolds for Tissue Engineering Applications BABT
Jaiswal,Amit.
The discipline of tissue engineering opens up the ways for repair and regenerate damaged organs and tissues. In the current work biomimetic nanofibrous scaffolds were fabricated by electrospinning. Poly-L-lactic acid (PLLA) was blended with collagen and gelatin to fabricate PLLLA-collagen and PLLA-gelatin fibrous scaffolds respectively. Pure PLLA and gelatin scaffolds served as controls. All the scaffolds displayed randomly oriented smooth fibers studied by SEM. Surface topography and roughness were studied by AFM and surface contact angle was also measured for all the fabricated scaffolds. Surface roughness was found to be higher in collagen and gelatin blended scaffolds in comparison to PLLA scaffold. Blending of collagen and gelatin reduced the surface...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Electrospinning; Biomimetic; Tissue Engineering.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100602
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Genetic variability in four fish species (Pimelodus maculatus, Prochilodus lineatus, Salminus brasiliensis and Steindachneridion scripta) from Uruguay River basin BABT
Ramella,Micheline Sandra; Kroth,Mariela Aparecida; Meurer,Samira; Nuñer,Alex Pires de Oliveira; Zaniboni Filho,Evoy; Arisi,Ana Carolina Maisonnave.
The genetic variability of four fish species (Pimelodus maculatus, Prochilodus lineatus, Salminus brasiliensis and Steindachneridion scripta) collected in the upper Uruguay River basin was analyzed using the RAPD technique. A total of 118 amplified fragments was obtained, 11 for P. maculatus, 29 for P. lineatus, 45 for S. brasiliensis and 33 for S. scripta. Amplified fragments with monomorphic profile were not found in the studied species, except for S. brasiliensis, which presented seven monomorphic bands for Saltinho population. All species showed high levels of genetic variability among individuals.
Tipo: Info:eu-repo/semantics/article Palavras-chave: RAPD; Pimelodus maculatus; Prochilodus lineatus; Salminus brasiliensis; Steindachneridion scripta.
Ano: 2006 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132006000500008
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Maize Productivity, Mycorrhizal Assessment, Chemical and Microbiological Soil Attributes Influenced by Maize-Forage Grasses Intercropping BABT
Méndez,Daniel Fernando Salas; Paula,Alessandra Monteiro de; Ramos,Maria Lucrécia Gerosa; Busato,Jader Galba.
Abstract Mycorrhizae are important components of agroecosystems and the diversification of crops stimulates the abundance of arbuscular mycorrhizal fungi and the participation of symbiosis in plant growth. This experiment examined mycorrhizal assessment, chemical and microbiological soil attributes in a maize-forage grasses intercropping compared to a maize-monocropping system. A complete randomized block design was used with crop systems installed under no-till management with three replicates, as follow: Maize (Zea mays L.); Panicum maximum Jacq. cv. Aruana; Urochloa humidicola (Rendle.) Schweickerdt.; Maize-P. maximum intercropping and Maize-U. humidicola intercropping. In 2015/2016 season, intercropping maize with Panicum maximum Jacq. cv. Aruana or...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cropping systems; Arbuscular mycorrhiza; Tropical soils; Grain crops.
Ano: 2019 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132019000100205
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Development of a Low Cost Bioprocess for Endotoxin Production by Bacillus thuringiensis var israelensis Intended for Biological Control of Aedes aegypti BABT
Soccol,Carlos Ricardo; Pollom,Teresinha E. V.; Fendrich,Ricardo Cancio; Prochmann,Fernando Alberto; Mohan,Radijiskumar; Blaskowski,Marita Maciel Moreira; Melo,André Luiz de Almeida; Carvalho,Cláudio Jose Barros de; Soccol,Vanete Thomaz.
Aedes aegypti is the vector of Dengue disease, responsible for 20,000 deaths/year worldwide. Bacillus thuringiensis var israelensis - Bti releases selective and effective toxins (crystal proteins) against A. aegypti larvae. We present a low cost bioprocess for toxin production, accomplished by a selected Brazilian strain Bti (BR-LPB01) and employment of low cost substrates. Soybean meal and sugarcane molasses lead to high toxic effectiveness after 2L bioreactor fermentation (LD50=26ng/mL), near to the reference strain IPS82 (LD50=17.3 ng/mL). The pH ranged between 5.8 and 7.0 during the exponential growth period and between 7.0 and 8.4 during the stationary phase, with low activity. Thus, control of foam and pH 7.0 were started and proved to be crucial for...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Biopesticide; BTI; Dengue Fever Disease; Soybean meal; Sugarcane molasses.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700017
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Bone-seeking therapeutic radiopharmaceuticals BABT
Srivastava,Suresh C..
Bone-seeking therapeutic radiopharmaceuticals are utilized on the basis of the radionuclide’s particulate emissions (primarily low to intermediate beta emission). The requirements therefore are different from those of bone imaging agents that consist mainly of short-lived single photon emitters. Lately, the therapeutic bone seeking radiopharmaceuticals have attained increasing importance due to their potential role in alleviating pain from osseous metastases in cancer patients, for the treatment of joint pain resulting from inflamed synovium (radiosynoviorthesis, or radiosynovectomy), or from various other forms of arthritic disease. There is, however, a paucity of published data on the bio-pharmacokinetics of these agents when used following intravenous...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Therapeutic radiopharmaceuticals; Bone seekers; Pain palliation; Bio-pharmacokinetics; Radiosynovectomy.
Ano: 2002 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132002000500007
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Recovery of ascorbic oxidoreductase from crude extract with an aqueous two-phase system in a perforated rotating disc contactor BABT
Porto,Ana Lúcia Figueiredo; Sarubbo,Leonie Asfora; Moreira,Keila Aparecida; Melo,Homero José Farias de; Lima-Filho,José Luís; Campos-Takaki,Galba Maria; Tambourgi,Elias Basile.
A continuous perforated rotating disc contactor was used to extract the enzyme ascorbic oxidoreductase (E.C.1.10.3.3) from crude extract of Curcubita maxima with an aqueous two-phase system of poly (ethylene glycol) and phosphate salts. The effect of dispersed phase velocity on either protein mass transfer coefficients or separation efficiency at 1, 2 and 3 mL/min was studied. An increase of the mass transfer coefficients was observed with the dispersed phase velocity, while the separation efficiency showed a small decrease with the increase of this parameter. The experimental results obtained during continuous extraction showed that the ascorbic oxidoreductase activity was partitioned preferentially into the salt-rich phase in all conditions studied. The...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Efficiency; Mass transfer coefficient; Perforated Rotating Disc contactor; Aqueous two-phase system; Ascorbic oxidoreductase.
Ano: 2004 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132004000500019
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Monitoring the nutritional and reproductive state of dairy cows through the presence of urea in milk BABT
Grande,Paula Adriana; Santos,Geraldo Tadeu dos; Ribeiro,Hilton; Damasceno,Júlio César; Alcalde,Claudete Regina; Barbosa,Orlando Rus; Horst,José Augusto; Santos,Fabio Seiji dos.
The effects of farm, parity (PO) and month of parturition on milk production, percentage of fat and protein, somatic cell count (SCC), and milk urea nitrogen (MUN) concentration were assessed on four specialized dairy farms using 120 cows. Also, the effects of MUN on gestation rate (GR) and artificial insemination rate (AIR) in early lactation were studied. The parameters of production and milk quality were similar among farms and were not influenced by the month of parturition. Farm D presented the highest MUN concentration. Concentration of MUN, percentages of fat and protein and SCC were not influenced by PO. Cows with a value of MUN between 10.1 and 13.0 mg/dL had the highest AIR and GR between days 55 and 70 postpartum and after 70 days in milk. These...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Dairy cattle; Gestation; Insemination; Milk urea nitrogen; Parturition order.
Ano: 2009 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132009000700032
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Experimental Design Applied to the Optimization and Partial Characterization of Pectin Liase from a Newly Isolated Penicillium brasilianum BABT
Zeni,Jamile; Gomes,Jonaína; Ambroszini,Éllin; Basso,Ana Paula; Toniazzo,Geciane; Valduga,Eunice.
Penicillium brasilianum was previously isolated from tea and identified by molecular biology technique. A Plackett-Burman design, followed by a complete second order design was used for the screening of most important factors and to maximize the pectin liase (PMGL) activity, respectively. The maximum PMGL activity by P. brasilianum achieved was 9.0 U/mL after 48 h of cultivation in a medium containing pectin (33.0 g/L), yeast extract (30.0 g/L) and potassium phosphate (2.0 g/L) at 30ºC, with a stirring rate of 180 rpm, initial pH 5.5 and 5x106spores/mL inoculum size. The kinetic evaluation in terms of substrate consumption demonstrated that the maximum production of PMGL was at 72 h, and 40% of the total organic carbon, 25% of the nitrogen, 88% of the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Penicillium brasilianum; Pectin liase; Experimental design technique.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000600908
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Effect of crude extract and its semi purified constituents from guaraná seeds [Paullinia cupana var. sorbilis (Mart.) lucke] on cognitive performance in Morris water maze in rats BABT
Otobone,Fernanda Jacques; Sanches,Andréia Conegero; Nagae,Rosangela Lika; Martins,Juliana Vanessa Colombo; Obici,Simoni; Mello,João Carlos Palazzo de; Audi,Elisabeth Aparecida.
The effect of crude lyophilized extract (EBPC) and the semi-purified constituents (EPA and EPB) of Paullinia cupana (guaraná) seeds long-term administered in rats by gavage at different doses was studied on cognitive behavior in rats. EBPC (30.0 mg/kg) and EPA (2.0 mg/kg), but not EPB (2.0 or 4.0 mg/kg) showed a smaller escape latency to find the emerged platform in Morris water maze test (MWMT), showing nootropic-like effect in normal rats, and in scopolamine induced amnesia rats compared to their controls (saline + 0.2% Tween 80) group. These extracts had no significant effect in open field test (OFT). Caffeine did alter escape latency in MWMT only in scopolamine induced amnesia rats and increased the crossings number in OFT, showing significant...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Cognitive effects; Guaraná; Morris water maze; Paullinia cupana.
Ano: 2005 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132005000600007
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Effects of fenoprofen on the labeling of blood constituents with technetium-99m, the morphology of red blood cells and the plasmid BABT
Pereira,Marcia de Oliveira; Rocha,Gabrielle de Souza; Lombardi,Simone dos Santos; Geller,Mauro; Pereira,Mário José; Santos-Filho,Sebastião David; Fonseca,Adenilson de Souza da; Bernardo-Filho,Mario.
The aim of this work was to evaluate the effect of fenoprofen on the labeling of blood constituents with technetium-99m, on the morphology of red blood cells and on the plasmid DNA. Blood samples from Wistar rats were incubated with fenoprofen and the assay of labeling of blood constituents with technetium-99m (99mTc) was performed. Blood cells, plasma, soluble and insoluble fractions of blood cells and plasma were separated. The radioactivity in each fraction was counted and percentage of incorporated radioactivity (%ATI) was determined. Blood smears were prepared, fixed, stained and the qualitative and quantitative morphology of the red blood cells (RBC) was evaluated. Plasmid (pBSK) was incubated with fenoprofen with stannous chloride, and agarose gel...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Technetium-99m; Blood; Morphology; Plasmid; Fenoprofen.
Ano: 2008 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132008000700022
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Effects of GDF-9 and FSH on mRNA expression for FSH-R, GDF-9 and BMPs in in vitro cultured goat preantral follicles BABT
Leitão,Cintia Camurça Fernandes; Costa,José Jackson do Nascimento; Brito,Ivina Rocha; Magalhães-Padilha,Déborah de Melo; Almeida,Anderson Pinto; Figueiredo,José Ricardo de; van den Hurk,Robert; Silva,José Roberto Viana.
The aim of the present study was to determine the role of GDF-9 and/or FSH on the growth and mRNA expression for FSH-R, GDF-9, and BMPs in goat secondary follicles after culture in vitro. Goat secondary follicles (~200µm) were isolated and cultured for six days in minimum essential medium (MEM) supplemented with GDF-9 (200 ng/mL), FSH (50 ng/mL) or both. At the beginning and end of culture, the follicular diameter was evaluated and compared. The levels of mRNA for GDF-9, FSH-R and BMPs -2, -4, -6, -7 and -15 in cultured follicles were quantified by real time PCR. The results showed that a significant increase of follicle diameter after six days when compared to day 0, but the presence of GDF-9 and FSH did not influence the follicular growth in comparison...
Tipo: Info:eu-repo/semantics/article Palavras-chave: BMPs; Follicles; GDF-9; In vitro; MRNA.
Ano: 2014 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132014000200007
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Tropical Fruit Pulps: Processing, Product Standardization and Main Control Parameters for Quality Assurance BABT
Silva,Carlos Eduardo de Farias; Abud,Ana Karla de Souza.
ABSTRACT Fruit pulp is the most basic food product obtained from fresh fruit processing. Fruit pulps can be cold stored for long periods of time, but they also can be used to fabricate juices, ice creams, sweets, jellies and yogurts. The exploitation of tropical fruits has leveraged the entire Brazilian fruit pulp sector due mainly to the high acceptance of their organoleptic properties and remarkable nutritional facts. However, several works published in the last decades have pointed out unfavorable conditions regarding the consumption of tropical fruit pulps. This negative scenario has been associated with unsatisfactory physico-chemical and microbiological parameters of fruits pulps as outcomes of little knowledge and improper management within the...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Processing; Standards of identity and quality; Vegetable product; Food safety.
Ano: 2017 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132017000100502
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Ovary maturation stages and oocyte features in three species of the neotropical fish Hemiodus (Müller, 1842) BABT
Brandão,Claudia Angelica da Silva; Valentim,Maria de Fátima Moraes; Caramaschi,Érica Pellegrini.
Macro- and microscopic ovarian features of Hemiodus microlepis, H. ternetzi and H. unimaculatus were analyzed. Based on these features we proposed an ovarian maturation scale. The nine stages of the preliminary macroscopic fieldwork scale were reduced to five after microscopic analysis of ovaries. The microscopic analysis indicated a group-synchronous oocyte development common to the three species that were characterized as iteroparous synchronous spawners with a total spawning type. The remarkable thickness of the zona radiata layer and the large size of vitellogenic oocytes of Hemiodus ternetzi distinguished this species from the others.
Tipo: Info:eu-repo/semantics/article Palavras-chave: Hemiodontid; Oocyte development; Zona radiata; Fish reproduction; Tocantins River.
Ano: 2003 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132003000300015
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The Role of Apollon Gene Silencing on Viablity and Radiosensitivity of Cervical Cancer Hela Cells BABT
Milani,Saeideh; Bandehpour,Mojgan; Sharifi,Zohreh; Kazemi,Bahram.
Cervical cancer is the second common cause of cancer deaths in women worldwide. Radioresistancy of cancer is a principal cause of treatment impairing. Inhibitor of apoptosis proteins (IAPs) widely block apoptosis against apoptotic stimuli, including current chemo- and radiation therapies. Apollon, a membrane of IAP, can support cells against apoptosis and is over expressed in some treatment-resistant cancer cells. The aim of this study was to evaluate the effects of apollon knockdown on induction of apoptosis and also its potential for enhancement of radiosensitvity on hela cells. plasmid encoding shRNA which has been confirmed its effect against apollon, transfected into hela cells. Consequent effects on the level of P53 , Bax and BAK analyzed by real...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Apollon; Apoptosis; Radiosensitivity; Cervical cancer.
Ano: 2016 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S1516-89132016000100320
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