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Bovine oocyte vitrification: Effect of ethylene glycol concentrations and meiotic stages

MAGNUSSON, Vanessa; FEITOSA, Weber Beringui; GOISSIS, Marcelo Demarchi; YAMADA, Claudia; TAVARES, Liliam Mara Trevisan; ORTIZ, Mayra Elena; ASSUMPCAO, D`Avila; VISINTIN, Jose Antonio
Fonte: ELSEVIER SCIENCE BV Publicador: ELSEVIER SCIENCE BV
Tipo: Artigo de Revista Científica
Português
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47.58359%
Success in oocyte cryopreservation is limited and several factors as cryoprotectant type or concentration and stage of oocyte meiotic maturation are involved. The aim of the present study was to evaluate the effect of maturation stage and ethylene glycol (EG) concentration on survival of bovine oocytes after vitrification. In experiment 1, kinetics of oocyte in vitro maturation (IVM) was evaluated. Germinal vesicle (GV), germinal vesicle breakdown (GVBD), metaphase I (MI), and metaphase II (MII) oocytes were found predominantly at 0, 0-10, 10-14, and 18-24 h of INK respectively. In experiment 2, in vitro embryo development after in vitro fertilization (IVF) of oocytes exposed to equilibrium (ES) and vitrification solution VS-1 (EG 30%), or VS-2 (EG 40%) at 0, 12 or 18 It of IVM was evaluated. Only blastocyst rate from oocytes vitrified in SV-2 after 18 h of IVM was different from control oocytes. Hatched blastocyst rates from oocytes vitrified in VS-1 after 12 and 18 h, and SV-2 after 18 h of IVM were different from unvitrified oocytes. In experiment 3, embryo development was examined after IVF of oocytes vitrified using VS-I or VS-2 at 0, 12 or 18 h of IVM. Rates of blastocyst development after vitrification of oocytes in VS-1 at each time interval were similar. However...

Effects of gonadotropin-exposed medium with high concentrations of progesterone and estradiol-17 beta on in vitro maturation of canine oocytes

VANNUCCHI, Camila Infantosi; FAUSTINO, Marcelo; MARQUES, Mariana Groke; NICHI, Marcilio; ASSUMPCAO, Mayra Elena Ortiz D`Avila; VISINTIN, Jose Antonio
Fonte: SPRINGER Publicador: SPRINGER
Tipo: Artigo de Revista Científica
Português
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47.51626%
During the process of maturation in the oviduct, canine oocytes in the germinal vesicle stage are exposed to decreasing levels of estradiol-17 beta and increasing levels of progesterone. However, hormone concentrations in the microenvironments in which they act are higher than serum concentrations. Therefore, the aim of the present study was to compare the meiotic competence of canine oocytes harvested from anestrous bitches in culture medium containing high concentrations (20 mu g ml(-1)) of estradiol-17 beta and/or progesterone in association to gonadotropins (luteinizing hormone and follicle-stimulating hormone) using three different maturation periods (48, 72, and 96 h). Oocytes were cultured in tissue culture medium (TCM-199) and arranged in four experimental groups: group control, group E2 (estradiol-17 beta), group P4 (progesterone), and group E2 + P4. Regardless of the maturation period, groups P4 and E2 + P4 presented statistically higher rate of germinal vesicle breakdown oocytes compared to the group control and group E2. There were no significant differences among groups on germinal vesicle, metaphase I, metaphase II, and degenerated or unidentifiable oocytes rates. The mean percentage of metaphase II oocytes was higher at 96 h when compared to 72 h. Results of the present research indicate no influence of estradiol-17 beta supplementation...

Efeito da pré-maturação sobre o desenvolvimento embrionário de oócitos submetidos à ativação partenogenética e transferência de núcleo; Effect of pre-maturation on embryo development of oocytes submitted to parthenogenetic activation and nuclear transfer

De Bem, Tiago Henrique Camara
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 03/06/2009 Português
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As taxas de produção embrionária tanto da FIV (30-40%) como da TN (23%) ainda estão aquém do esperado. Desta forma, a pré-maturação com inibidores do ciclo celular é uma das alternativas que vem sendo estudada para aumentar a competência dos oócitos utilizados na PIV, na tentativa de otimizar o sucesso das biotécnicas. Sabe-se que neurotrofinas desempenham funções no sistema reprodutor. O BDNF é um exemplo de neurotrofina que parece estar relacionada com a maturação dos oócitos. Desta forma, o objetivo deste trabalho foi de aperfeiçoar a pré-maturação e a maturação in vitro de oócitos bovinos submetidos posteriormente à ativação partenogenética, visando seu uso na biotécnica de transferência de núcleo. Oócitos bovinos foram submetidos à maturação na presença (MIV/BD) ou ausência (MIV) de BDNF ou pré-maturados com BLI e suplementados (BL/BD) ou não (BL) com a neurotrofina. Posteriormente foram avaliados quanto à taxa de maturação (metáfase II), ativação (formação de prónúcleo) e desenvolvimento embrionário (produção e qualidade dos blastocistos). Não houve diferença (P>0,05) entre taxas de MII após a maturação com ou sem BDNF. Porém, o grupo pré-maturado e suplementado (BL/BD n=73; 91...

Influência do hormônio folículo estimulante na via da óxido nítrico sintase em complexos cumulus-oócitos bovinos.; Influency of folicular stimulant hormon on the nitric oxide pathway in bovines cumulus-oocyte complex.

Pires, Pedro Ratto Lisboa
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 17/12/2010 Português
Relevância na Pesquisa
47.49489%
O óxido nítrico (NO) é um mensageiro químico gerado pela atividade da enzima óxido nítrico sintase (NOS) a qual foi detectada em vários órgãos incluídos os do sistema reprodutor e parece estar envolvido na maturação oocitária. No entanto, há poucos estudos sobre o papel desse sistema em oócitos da espécie bovina. Sabe-se que o NO atua pela via da guanilato ciclase (GC) estimulando a produção do nucleotídeo GMPc, que por sua vez é capaz de influenciar nos níveis de outro nucleotídeo (AMPc) via fosfodiesterases (PDE). O AMPc é um importante elemento da via de sinalização do FSH nos complexos cumulus-oócitos e no controle da maturação oocitária. O objetivo do presente projeto foi investigar a influência do FSH na via do NOS/NO e seus componentes em oócitos bovinos maturados in vitro e o envolvimento das células do cumulus (CC) na via de sinalização. Para tanto, complexos cumulus-oócito (CCO) e oócitos desnudos (OD - maturados sem células do cumulus) foram maturados in vitro por 24h na presença ou ausência de FSH. As amostras foram avaliadas quanto a: 1) taxa da maturação nuclear; 2) níveis de produção de NO; 3) níveis de AMPc e GMPc; 4) abundância relativa de RNAm de NOS2, PDE5A, PDE6C, PKG1...

Ultrastructural Morphology and Nuclear Maturation Rates of Immature Equine Oocytes Vitrified with Different Solutions and Exposure Times

Curcio, Bruna R.; Gastal, Melba O.; Pereira, Gabriel R.; Corcini, Carine D.; Landim-Alvarenga, Fernanda C.; Barros, Severo S.; Nogueira, Carlos E. W.; Deschamps, Joao Carlos; Gastal, Eduardo L.
Fonte: Elsevier B.V. Publicador: Elsevier B.V.
Tipo: Artigo de Revista Científica Formato: 632-640
Português
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Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES); Ultrastructural morphological injuries and maturation rates were investigated in equine oocytes exposed to vitrification solutions (VS) containing synthetic ice blockers (SIBs) during different exposure times. In experiment 1, compact cumulus-oocyte complexes (COCs; n = 30) were randomly allocated to treatments: (1) fresh fixed (control); (2) VS-1 (1.4 M dimethyl sulfoxide [DMSO] + 1.8 M ethylene glycol LEG] + 1% SIB) for 3 minutes of equilibrium time and VS-2 (2.8 M DMSO + 3.6 M EG + 0.6 M sucrose + 1% SIB) for 1 minute (Eq-long); and (3) VS-1 for 1.5 minutes and VS-2 for 30 seconds (Eq-short). In experiment 2, compact (n = 248) and expanded (n = 264) COCs were evenly distributed to the following treatments: (1) immediate maturation in vitro (control); (2) vitrification using the Eq-short protocol as in experiment 1; and (3) vitrification using a stock solution containing 2.8 M formamide, 2.8 M DMSO, 2.7 M EG, 7% polyvinylpyrrolidone, and 1% SIB (Eq-short-mod). More (P <.02) oocytes with normal ultrastructural morphology were seen in fresh control and Eq-short groups than in Eq-long group. Metaphase-II (MIL) rates were higher (P <.05) for oocytes with expanded cumulus than compact cumulus in the control group...

Apical and basolateral coated pits of MDCK cells differ in their rates of maturation into coated vesicles, but not in the ability to distinguish between mutant hemagglutinin proteins with different internalization signals

Fonte: The Rockefeller University Press Publicador: The Rockefeller University Press
Tipo: Artigo de Revista Científica
Publicado em 01/06/1995 Português
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In polarized epithelial MDCK cells, all known endogenous endocytic receptors are found on the basolateral domain. The influenza virus hemagglutinin (HA) which is normally sorted to the apical plasma membrane, can be converted to a basolateral protein by specific mutations in its short cytoplasmic domain that also create internalization signals. For some of these mutations, sorting to the basolateral surface is incomplete, allowing internalization of two proteins that differ by a single amino acid of the internalization signal to be compared at both the apical and basolateral surfaces of MDCK cells. The rates of internalization of HA-Y543 and HA-Y543,R546 from the basolateral surface of polarized MDCK cells resembled those in nonpolarized cells, whereas their rates of internalization from the apical cell surface were fivefold slower. However, HA-Y543,R546 was internalized approximately threefold faster than HA-Y543 at both membrane domains, indicating that apical endocytic pits in polarized MDCK cells retained the ability to discriminate between different internalization signals. Slower internalization from the apical surface could not be explained by a limiting number of coated pits: apical membrane contained 0.7 as many coated pits per cell cross-section as did basolateral membranes. 10-14% of HA-Y543 at the apical surface of polarized MDCK cells was found in coated pits...

Subunits fold at position-dependent rates during maturation of a eukaryotic RNA virus

Matsui, Tsutomu; Lander, Gabriel C.; Khayat, Reza; Johnson, John E.
Fonte: National Academy of Sciences Publicador: National Academy of Sciences
Tipo: Artigo de Revista Científica
Português
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47.469478%
Effective antiviral agents are difficult to develop because of the close relationship between the cell biology of the virus and host. However, viral capsid maturation, the in vivo process where the particle transitions from a noninfectious provirion to an infectious virion, is an ideal process to interrupt because the provirion is usually fragile and the conversion to the virion often involves large conformational changes and autocatalytic chemistry that can be hampered by small molecules. The Nudaurelia capensis ω virus (NωV) is one of the few eukaryotic viruses where this process can be investigated in vitro with a variety of biophysical methods, allowing fundamental chemical and structural principles of the maturation to be established. It has a T = 4 quasi-equivalent capsid with a dramatic maturation pathway that includes a particle size reduction of 100 Å and an autocatalytic cleavage. Here we use cryo-EM and difference maps, computed at three time points following maturation initiation, to show that regions of NωV subunit folding are maturation dependent and occur at rates determined by their quasi-equivalent position in the capsid, explaining the unusual kinetics of the maturation cleavage. This study shows that folding is rapid and peptide chain self-cleavage occurs early for subunits adjacent to 3-fold and 5-fold icosahedral symmetry elements and that folding is slower in regions where molecular switches are required for the formation of the proper interfacial contacts. The results connect viral maturation to the well-studied assembly-dependent folding that occurs in the formation of cellular complexes.

Developmental Acquisition of a Rapid Calcium-Regulated Vesicle Supply Allows Sustained High Rates of Exocytosis in Auditory Hair Cells

Levic, Snezana; Bouleau, Yohan; Dulon, Didier
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Publicado em 06/10/2011 Português
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56.721357%
Auditory hair cells (HCs) have the remarkable property to indefinitely sustain high rates of synaptic vesicle release during ongoing sound stimulation. The mechanisms of vesicle supply that allow such indefatigable exocytosis at the ribbon active zone remain largely unknown. To address this issue, we characterized the kinetics of vesicle recruitment and release in developing chick auditory HCs. Experiments were done using the intact chick basilar papilla from E10 (embryonic day 10) to P2 (two days post-hatch) by monitoring changes in membrane capacitance and Ca2+ currents during various voltage stimulations. Compared to immature pre-hearing HCs (E10-E12), mature post-hearing HCs (E18-P2) can steadily mobilize a larger readily releasable pool (RRP) of vesicles with faster kinetics and higher Ca2+ efficiency. As assessed by varying the inter-pulse interval of a 100 ms paired-pulse depolarization protocol, the kinetics of RRP replenishment were found much faster in mature HCs. Unlike mature HCs, exocytosis in immature HCs showed large depression during repetitive stimulations. Remarkably, when the intracellular concentration of EGTA was raised from 0.5 to 2 mM, the paired-pulse depression level remained unchanged in immature HCs but was drastically increased in mature HCs...

Quantitative Genetics of Body Size and Timing of Maturation in Two Nine-Spined Stickleback (Pungitius pungitius) Populations

Shimada, Yukinori; Shikano, Takahito; Kuparinen, Anna; Gonda, Abigél; Leinonen, Tuomas; Merilä, Juha
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Publicado em 14/12/2011 Português
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Due to its influence on body size, timing of maturation is an important life-history trait in ectotherms with indeterminate growth. Comparison of patterns of growth and maturation within and between two populations (giant vs. normal sized) of nine-spined sticklebacks (Pungitius pungitius) in a breeding experiment revealed that the difference in mean adult body size between the populations is caused by differences in timing of maturation, and not by differential growth rates. The fish in small-sized population matured earlier than those from large-sized population, and maturation was accompanied by a reduction in growth rate in the small-sized population. Males matured earlier and at smaller size than females, and the fish that were immature at the end of the experiment were larger than those that had already matured. Throughout the experimental period, body size in both populations was heritable (h2 = 0.10–0.64), as was the timing of maturation in the small-sized population (h2 = 0.13–0.16). There was a significant positive genetic correlation between body size and timing of maturation at 140 DAH, but not earlier (at 80 or 110 DAH). Comparison of observed body size divergence between the populations revealed that QST exceeded FST at older ages...

Differential Growth and Development of the Upper and Lower Human Thorax

Bastir, Markus; García Martínez, Daniel; Recheis, Wolfgang; Barash, Alon; Coquerelle, Michael; Rios, Luis; Peña-Melián, Ángel; García Río, Francisco; O’Higgins, Paul
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Publicado em 20/09/2013 Português
Relevância na Pesquisa
56.723613%
The difficulties in quantifying the 3D form and spatial relationships of the skeletal components of the ribcage present a barrier to studies of the growth of the thoracic skeleton. Thus, most studies to date have relied on traditional measurements such as distances and indices from single or few ribs. It is currently known that adult-like thoracic shape is achieved early, by the end of the second postnatal year, with the circular cross-section of the newborn thorax transforming into the ovoid shape of adults; and that the ribs become inclined such that their anterior borders come to lie inferior to their posterior. Here we present a study that revisits growth changes using geometric morphometrics applied to extensive landmark data taken from the ribcage. We digitized 402 (semi) landmarks on 3D reconstructions to assess growth changes in 27 computed tomography-scanned modern humans representing newborns to adults of both sexes. Our analyses show a curved ontogenetic trajectory, resulting from different ontogenetic growth allometries of upper and lower thoracic units. Adult thoracic morphology is achieved later than predicted, by diverse modifications in different anatomical regions during different ontogenetic stages. Besides a marked increase in antero-posterior dimensions...

Demographic Structure, Sex Ratio and Growth Rates of Southern Bluefin Tuna (Thunnus maccoyii) on the Spawning Ground

Farley, Jessica H.; Eveson, J. Paige; Davis, Tim L. O.; Andamari, Retno; Proctor, Craig H.; Nugraha, Budi; Davies, Campbell R.
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Publicado em 05/05/2014 Português
Relevância na Pesquisa
56.722026%
The demographics of the southern bluefin tuna (SBT) Thunnus maccoyii spawning stock were examined through a large-scale monitoring program of the Indonesian longline catch on the spawning ground between 1995 and 2012. The size and age structure of the spawning population has undergone significant changes since monitoring began. There has been a reduction in the relative abundance of larger/older SBT in the catch since the early 2000s, and a corresponding decrease in mean length and age, but there was no evidence of a significant truncation of the age distribution. Pulses of young SBT appear in the catches in the early- and mid-2000s and may be the first evidence of increased recruitment into the spawning stock since 1995. Fish in these two recruitment pulses were spawned around 1991 and 1997. Size-related variations in sex ratio were also observed with female bias for fish less than 170 cm FL and male bias for fish greater than 170 cm FL. This trend of increasing proportion of males with size above 170 cm FL is likely to be related to sexual dimorphism in growth rates as male length-at-age is greater than that for females after age 10 years. Mean length-at-age of fish aged 8–10 years was greater for both males and females on the spawning ground than off the spawning ground...

Quantifying the hemodynamic characteristics of an arteriovenous fistula used for hemodialysis

Browne, Leonard D
Fonte: University of Limerick Publicador: University of Limerick
Tipo: info:eu-repo/semantics/doctoralThesis; all_ul_research; ul_published_reviewed; ul_theses_dissertations
Português
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47.591455%
peer-reviewed; Chronic kidney disease refers to an irreversible, progressive reduction of the renal function of kidneys. When there is a significant loss of renal function a replacement therapy is sought. A kidney transplant is the optimal replacement. However, the demand for kidney transplantation exceeds the supply of transplantable organs and most patients will never receive a kidney transplant. As a consequence, these patients are often referred to hemodialysis to filter blood. To facilitate efficient and adequate dialysis an access capable of supplying high flow rates is required. An arteriovenous fistula is the access modality of choice for patients requiring hemodialysis. This access is formed from the anastomosis of an artery and vein which generates the high flow rates needed for efficient dialysis. Despite the preference of this access, the modality suffers from drastically poor primary and secondary patency rates which necessitate urgent improvement. Following the creation of the access a period of time is required to ensure adequate remodelling and maturation of the fistula occurs prior to cannulation. Non-maturation contributes significantly to the dismal patency rates of this access. Impaired outward remodelling and aggressive intimal hyperplasia are both considered to contribute to fistula non-maturation and the abnormal hemodynamics arising from fistula creation are believed to provide a stimulus for both facets of remodelling. Increasing an understanding of this relationship forms the primary focus of this dissertation. The primary aim of this thesis is to accurately characterise the hemodynamics of an arteriovenous fistula using experimental and numerical models. An unsteady incompressible Navier Stokes solver strategy was implemented and validated as a reliable computational approach to accurately estimate the pressure drop across a representative fistula. Both in-vitro and numerical models demonstrated a quadratic relationship between pressure drop and flow rate across the anastomosis. Instabilities were found to develop within the anastomosis and generated high frequency oscillations of the pressure drop signal. These instabilities were most prevalent during the end of the systole and throughout diastole phase of the cycle. Direct in-vivo measurements of the pressure distribution within two brachiocephalic fistulae were obtained intra operatively to characterise the hemodynamics of a fistula and provide a further benchmark for numerical solutions. A continuous pressure drop across these fistulae was observed and coincided with the presence of perivascular vibrations known as thrills. The representative fistula model was scaled to match characteristics of a patient case and a favourable comparison was observed between the pressure distribution of the in-vivo and numerical case. High frequency oscillations of the numerical pressure drop signal were observed and the flow demonstrated an unstable behaviour. The numerical strategy was applied to a reconstructed case of a brachiocephalic fistula to assess the influence of hemodynamic parameters on remodelling. Outward remodelling over time was found to reduce the level of shear stress over time within the vein. However...

Factors affecting the developmental competence of pig oocytes matured in vitro.

Bagg, Melanie Anna
Fonte: Universidade de Adelaide Publicador: Universidade de Adelaide
Tipo: Tese de Doutorado
Publicado em //2007 Português
Relevância na Pesquisa
47.588203%
Pre-pubertal pig oocytes possess lower developmental competence than those from adult pigs following in vitro maturation (IVM). Previous studies have demonstrated that exposure of pre-pubertal oocytes to 1 mM dibutyryl cAMP (dbcAMP), a membrane permeable cyclic adenosine monophosphate (cAMP) analogue, for the first 20 h of IVM improves the rate of blastocyst development. Developmental competence of in vitro matured pig oocytes has been reported to increase with increasing follicle size. In this thesis, experiments were carried out using pre-pubertal and adult pig oocytes to investigate the relationship between donor age, intra-oocyte cAMP level and follicle size in terms of oocyte maturation and developmental competence. These experiments demonstrated that, while ovarian, follicular and oocyte morphology are immediately altered with the onset of puberty, pre-pubertal oocytes must be exposed to more than the first oestrous cycle to achieve improved developmental competence in vitro. Later experiments demonstrated that pre-pubertal oocytes accumulate less cAMP during IVM, undergo more rapid meiotic progression and display reduced rates of blastocyst development compared to in vitro matured adult oocytes. Treatment with dbcAMP for 22 h IVM increased the cAMP content of pre-pubertal oocytes...

Oxygen concentration during mouse oocyte in vitro maturation affects embryo and fetal development

Banwell, K.; Lane, M.; Russell, D.; Kind, K.; Thompson, J.
Fonte: Oxford Univ Press Publicador: Oxford Univ Press
Tipo: Artigo de Revista Científica
Publicado em //2007 Português
Relevância na Pesquisa
47.51156%
BACKGROUND: Little is known of how the oxygen environment in the ovarian follicle affects oocyte and embryo development, but this has an important impact on the conditions used for in vitro maturation (IVM) of oocytes. We investigated the effect of varying oxygen concentrations during IVM on subsequent pre and post-implantation development. METHODS: IVM of mouse cumulus-oocyte complexes (COCs) was performed under 2, 5, 10 or 20% O(2) (6% CO(2), balance N(2)). In vivo-matured COCs were collected post ovulation. Embryos were generated by IVF and culture. Blastocyst development, cell number and apoptosis were assessed, and fetal and placental outcomes analysed following embryo transfer at day 18 of pregnancy. RESULTS: Oxygen concentration during IVM did not affect oocyte maturation or subsequent fertilization, cleavage and blastocyst development rates. Maturation of oocytes under 2% O(2) increased blastocyst trophectoderm cell number compared with all groups and numbers at 5% were higher than 20% (both P < 0.05). Percentage of apoptotic cells was increased in blastocysts developed from 2% O(2)-matured oocytes, compared with maturation at 5% O(2) or in vivo (P < 0.05). Rates of embryo implantation and development into a viable fetus were not altered by IVM oxygen. However...

Differential growth and development of the upper and lower human thorax

Bastir, Markus; García Martínez, Daniel; Recheis, Wolfgang A.; Barash, Alon; Coquerelle, Michael; Ríos, Luis; Peña-Melián, Ángel L.; García-Rio, Franciso; O'Higgins, Paul
Fonte: Public Library of Science Publicador: Public Library of Science
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
56.723613%
The difficulties in quantifying the 3D form and spatial relationships of the skeletal components of the ribcage present a barrier to studies of the growth of the thoracic skeleton. Thus, most studies to date have relied on traditional measurements such as distances and indices from single or few ribs. It is currently known that adult-like thoracic shape is achieved early, by the end of the second postnatal year, with the circular cross-section of the newborn thorax transforming into the ovoid shape of adults; and that the ribs become inclined such that their anterior borders come to lie inferior to their posterior. Here we present a study that revisits growth changes using geometric morphometrics applied to extensive landmark data taken from the ribcage. We digitized 402 (semi) landmarks on 3D reconstructions to assess growth changes in 27 computed tomography-scanned modern humans representing newborns to adults of both sexes. Our analyses show a curved ontogenetic trajectory, resulting from different ontogenetic growth allometries of upper and lower thoracic units. Adult thoracic morphology is achieved later than predicted, by diverse modifications in different anatomical regions during different ontogenetic stages. Besides a marked increase in antero-posterior dimensions...

Effect of Hormonal Supplementation Periods and In Vitro Maturation Media on Developmental Competence of Pig Oocytes

LIMA, Alecssandra Sobreira de; MARQUES, Mariana Groke; NASCIMENTO, Anibal Ballarotti; ASSUMPCAO, Mayra Elena Ortiz D'Avila; VISITIN, Jose Antonio
Fonte: UNIV FED RIO GRANDE DO SUL Publicador: UNIV FED RIO GRANDE DO SUL
Tipo: Artigo de Revista Científica
Português
Relevância na Pesquisa
47.823926%
Background: The oocyte ability to undergo successful fertilization, cleavage and embryonic development depends on meiotic maturation and developmental competence acquisition. In vitro maturation (IVM) protocols currently use eCG, hCG or a combination of both, the effect of these gonadotrophins during IVM and subsequent embryonic development is still controversial. Several media have been used for IVM of porcine oocytes: TCM199, Whitten's and NCSU23 have also been shown to support pig oocyte IVM. This study was designed to determine the effect of hormonal supplementation period and maturation media during in vitro maturation of pig oocytes (1) and subsequent embryonic development (2). Materials, Methods & Results: Oocytes with intact cumulus oophurus layers and homogeneous cytoplasm were collected from prebubertal gilts. IVM was subjected in NCSU23, TCM199 or Whitten's media supplemented with 10 IU/mL eCG and 10 IU/mL hCG for the first 24 or 48 h of IVM. In each replicate the oocytes were fixed every 4 h from 32 to 48 h IVM or the past 48 h after IVM, oocytes were fertilized in vitro in mTBM medium for six hours and cultured in NCSU23 medium for nine days. Cleavage, blastocyst and hatching rates were evaluated at 48 h (day 2), 168 h (day 7) and 216 h (day 9)...

Índices de maturação, ponto de colheita e padrão respiratório de goiabas 'Kumagai' e 'Paluma'. ; Maturation rates, harvest point and respiratory standard of the ‘kumagai’ and ‘paluma’ guavas.

Cavalini, Flavia Cristina
Fonte: Biblioteca Digitais de Teses e Dissertações da USP Publicador: Biblioteca Digitais de Teses e Dissertações da USP
Tipo: Dissertação de Mestrado Formato: application/pdf
Publicado em 21/06/2004 Português
Relevância na Pesquisa
47.71286%
A goiaba é uma fruta altamente perecível, e o conhecimento de sua fisiologia pós-colheita é fundamental para o emprego adequado de tecnologias, visando aumentar o período de conservação. O presente trabalho foi realizado com os objetivos de determinar índices de maturação; verificar a influência dos estádios de maturação na qualidade pós-colheita e determinar o padrão respiratório para goiabas ‘Kumagai’ e ‘Paluma’. Primeiramente determinaram-se os índices de maturação e a influência de cinco estádios de maturação na qualidade pós-colheita dos frutos. Os frutos foram selecionados em cinco estádios de maturação segundo a cor da casca: Estádio 1: cor da casca verde-escura; Estádio 2: quebra da cor verde; Estádio 3: início da coloração amarela da casca; Estádio 4: cor da casca parcialmente amarela; Estádio 5: frutos com cor da casca totalmente amarela, em seguida, foram armazenados em câmara a 25 + 2ºC e 80-90% UR e avaliados quanto às mudanças físico-químicas e qualidade sensorial. Posteriormente determinou-se o padrão respiratório dos frutos, analisando-se a atividade respiratória, a produção de etileno e as mudanças físico-químicas após a colheita para os estádios 1, 2 e 3. A cor da casca e a firmeza foram consideradas os melhores índices de maturação para ambas as variedades. A variedade Paluma também apresentou o ratio como um bom índice de maturação. As variáveis físico-químicas apresentaram pouca variação entre os estádios de maturação após o amadurecimento...

Absolute rates of protein synthesis during meiotic maturation of mammalian oocytes in vitro.

Schultz, R M; LaMarca, M J; Wassarman, P M
Fonte: PubMed Publicador: PubMed
Tipo: Artigo de Revista Científica
Publicado em /09/1978 Português
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Measurements of the rates of incorporation of [35S]methionine into protein and the specific activities of endogenous free methionine pools have been used to calculate the absolute rates of protein synthesis in mouse oocytes during spontaneous meiotic maturation in vitro. Fluorodinitro[3H]benzene was used to determine the specific activity of the oocyte's free methionine pool. It was found that the absolute rate of protein synthesis decreased from 43 to 31 pg/hr per oocyte during meiotic progression from dictyate to metaphase II (meiotic maturation), while the size of the intracellular free methionine pool decreased from 61 to 35 fmol per oocyte during the same period. Comparable measurements made on ovulated mouse oocytes that had undergone meiotic maturation in vivo strongly suggest that the decrease in the absolute rate of protein synthesis observed during meiotic maturation in vitro is physiologically significant. An alternative method that depends upon differential expansion of the oocyte's endogenous methionine pool was also used to determine absolute rates of protein synthesis. The results of these experiments are in excellent agreement with those obtained by using fluorodinitro[3H]benzene, indicating that the oocyte's free methionine pool is not compartmentalized.

Maturação e desenvolvimento embrionário in vitro de oócitos bovinos após bloqueio da meiose com inibidores de MPF; In vitro maturation and embryo development of bovine oocytes after meiosis blockage with MPF inhibitors

Marques, Mariana Groke; Mello, Marco Roberto Bourg de; Tavares, Liliam Mara Trevisan; Nicacio, Alessandra Corallo; Assumpção, Mayra Elena Ortiz D'Avila; Visintin, José Antonio
Fonte: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia Publicador: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; ; ; ; ; ; Formato: application/pdf
Publicado em 01/01/2011 Português
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O objetivo deste estudo foi avaliar a maturação e o desenvolvimento embrionário após a fecundação in vitro de oócitos bovinos que tiveram a maturação bloqueada com Butirolactona I e Roscovitina em meio de pré-maturação suplementado com soro fetal bovino (SFB). Oócitos foram divididos em 4 grupos: Controle 0 hora, Controle (maturação por 24 horas), Butirolactona I (bloqueio da maturação com 150µM de Butirolactona I por 24 horas, seguido de 24 horas de maturação) e Roscovitina (bloqueio da maturação com 50µM de Roscovitina por 24 horas, seguido de 24 horas de maturação). Para avaliar a maturação nuclear, os oócitos foram fixados e corados em aceto orceína. Parte dos oócitos dos grupos Controle 24 horas, Roscovitina e Butirolactona I após o período de maturação, foi fecundado in vitro. O desenvolvimento embrionário foi avaliado pelos índices de clivagem (D3) e formação de blastocistos (D7). Oócitos do grupo Butirolactona I apresentaram índices de Vesícula Germinativa após o bloqueio e de Metáfase 2 após a maturação semelhantes ao dos grupos Controle 0 hora e Controle, respectivamente. Por outro lado, a Roscovitina apresentou menores índices de Vesícula Germinativa e Metáfase 2. Os grupos Controle e Butirolactona I apresentaram maiores índices de clivagens. O grupo Controle apresentou maior produção de blastocistos que o Roscovitina e não diferiu do grupo Butirolactona I. Conclui-se que a Butiroloactona I pode ser utilizada no sistema de pré-maturação em meio contendo SFB...

Cinética da maturação nuclear in vitro de oócitos bubalinos; Nulear maturation kinetcs in vitro of buffalo oocytes

Santos, Simone do Socorro Damasceno; Dantas, Jedina Kato; Miranda, Moysés dos Santos; Biondi, Flávia Costa; Ohashi, Otávio Mitio
Fonte: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia Publicador: Universidade de São Paulo. Faculdade de Medicina Veterinária e Zootecnia
Tipo: info:eu-repo/semantics/article; info:eu-repo/semantics/publishedVersion; ; ; ; ; ; Formato: application/pdf
Publicado em 01/01/2002 Português
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Com o objetivo de avaliar a cinética da maturação in vitro de oócitos bubalinos, foram cultivados 1.619 oócitos com cumulus oophorus compacto em quatro diferentes tratamentos: T1 - TCM 199 com 10% de Soro Fetal Bovino (SFB) (meio base); T2 - Meio base e células da granulosa (CG); T3 - Meio base, CG, 10 UI de gonadotrofina coriônica equina (eCG) e 10 UI de gonadotrofina coriônica humana (hCG); T4 - Meio base, 10 UI de eCG, 10UI de hCG e 1 µg de 17b-estradiol / ml, em tempos de 14 a 17h, 20 a 22h, 23 a 25h, 26 a 28h, 29h e 32h de cultivo em estufa de CO2 a 5% e temperatura de 38,5ºC. No tratamento 1, observou-se que 81,83%, 69,61%, 65,88%, 60,28%, 64,98% e 67,67% dos ovócitos reiniciaram a meiose. No tratamento 2, as taxas de reinicio da meiose foram de 76,06%, 72,06%, 69,78%, 68,12%,74,97% e 89,96%, no tratamento 3 estes percentuais foram de 83,59%, 78,86%, 77,49%, 82,4%, 72,92% e 81,06% e com o tratamento 4 os índices foram de 95,0%, 91,82%, 93, 17%, 91,65%, 92,09% e 74,99% nos tempos de 14 a 17h, 20 a 22h, 23 a 25h, 26 a 28h, 29h e 32h de cultivo, respectivamente, podendo-se concluir que o meio com maior suplementação proporciona maior taxa de reinício da meiose em menor tempo, assim como o aumento no tempo de cultivo pode levar a maiores índices de degeneração.; To evaluate the kinetics of the in vitro maturation of buffalo oocytes...