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1.
Biol Reprod ; 106(4): 814-822, 2022 04 26.
Artículo en Inglés | MEDLINE | ID: mdl-35040958

RESUMEN

Mature granulated trophoblast binucleate cells (BNC) have been found in all ruminant placentas examined histologically so far. BNC are normally fairly evenly distributed throughout the fetal villus and all their granules contain a similar variety of hormones and pregnancy associated glycoproteins (PAGs). Only the Giraffe is reported to show a different BNC protein expression, this paper is designed to investigate that. Gold labelled Lectin histochemistry and protein immunocytochemistry were used on deplasticised 1 µm sections of a wide variety of ruminant placentomes with a wide range of antibodies and lectins. In the Giraffe placentomes, even though the lectin histochemistry shows an even distribution of BNC throughout the trophoblast of the placental villi, the protein expression in the BNC granules is limited to the BNC either in the apex or the base of the villi. Placental lactogens and Prolactin (PRL) are present only in basally situated BNC: PAGs only in the apical BNC. PRL is only found in the Giraffe BNC which react with many fewer of the wide range of antibodies used here to investigate the uniformity of protein expression in ruminant BNC. The possible relevance of these differences to ruminant function and evolution is considered to provide a further example of the versatility of the BNC system.


Asunto(s)
Jirafas , Placenta , Animales , Femenino , Lectinas/metabolismo , Placenta/metabolismo , Embarazo , Prolactina/metabolismo , Rumiantes/metabolismo , Trofoblastos/metabolismo
2.
Int J Obes (Lond) ; 42(3): 507-517, 2018 03.
Artículo en Inglés | MEDLINE | ID: mdl-28901330

RESUMEN

BACKGROUND/OBJECTIVES: Dieting is a popular yet often ineffective way to lower body weight, as the majority of people regain most of their pre-dieting weights in a relatively short time. The underlying molecular mechanisms driving weight regain and the increased risk for metabolic disease are still incompletely understood. Here we investigate the molecular alterations inherited from a history of obesity. METHODS: In our model, male high-fat diet (HFD)-fed obese C57BL/6J mice were switched to a low caloric chow diet, resulting in a decline of body weight to that of lean mice. We measured body composition, as well as metrics of glucose, insulin and lipid homeostasis. This was accompanied by histological and gene expression analysis of adipose tissue and liver to assess adipose tissue inflammation and hepatosteatosis. Moreover, acute hypothalamic response to (re-) exposure to HFD was assessed by qPCR. RESULTS & CONCLUSIONS: Within 7 weeks after diet switch, most obesity-associated phenotypes, such as body mass, glucose intolerance and blood metabolite levels were reversed. However, hepatic inflammation, hepatic steatosis as well as hypertrophy and inflammation of perigonadal, but not subcutaneous, adipocytes persisted in formerly obese mice. Transcriptional profiling of liver and perigonadal fat revealed an upregulation of pathways associated with immune function and cellularity. Thus, we show that weight reduction leaves signs of inflammation in liver and perigonadal fat, indicating that persisting proinflammatory signals in liver and adipose tissue could contribute to an increased risk of formerly obese subjects to develop the metabolic syndrome upon recurring weight gain.


Asunto(s)
Tejido Adiposo/metabolismo , Inflamación/metabolismo , Hígado/metabolismo , Obesidad/metabolismo , Pérdida de Peso/fisiología , Tejido Adiposo/química , Animales , Biomarcadores/análisis , Restricción Calórica , Hígado Graso/metabolismo , Hígado/química , Masculino , Ratones , Ratones Endogámicos C57BL , Obesidad/dietoterapia
3.
Reprod Domest Anim ; 51(2): 282-6, 2016 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-26936628

RESUMEN

Plasma concentrations of PAG-1 are used for pregnancy diagnosis and as a marker of placental/foetal well-being, while those of PAG-2 may be an indicator of abortion risk in Neospora caninum-infected cows. Studies have shown that N. caninum infection modifies PAG-1 and PAG-2 patterns in maternal blood plasma. However, no prior work has examined the effects of N. caninum infection on concentrations of PAGs in foetal fluids. In this study, PAG-1, PAG-2 and pH levels were determined in the amniotic and allantoic fluids of foetuses collected at 152 days of gestation from control uninfected dams and from dams experimentally infected with N. caninum on Day 110 of gestation. Foetal fluids from infected foetuses had significantly higher PAG-2 concentrations (p = 0.026) and pH values (p = 0.02) than fluids from non-infected foetuses. In infected foetuses, significantly higher concentrations of PAG-1 (p < 0.001) and PAG-2 (p < 0.001) were detected in fluid samples showing antibodies against N. caninum than those without antibodies. Moreover, pH values were significantly higher (p = 0.011) in foetal fluid samples with antibodies than in samples from non-infected foetuses. In conclusion, this is the first report on the effect of N. caninum infection on PAG levels in foetal fluids. Our results indicate that following the experimental infection of dams with N. caninum on Day 110 of gestation, foetal fluids collected from the infected foetuses of these dams featured higher PAG-1 and PAG-2 levels and pH values than fluids from non-infected controls, provided that the samples tested showed the presence of antibodies. The clinical implications of these findings are that following infection with N. caninum, most cows will experience some level of placental damage and that this injury correlates with foetal fluid PAG levels and pH.


Asunto(s)
Líquido Amniótico/química , Enfermedades de los Bovinos/metabolismo , Coccidiosis/veterinaria , Neospora , Complicaciones Parasitarias del Embarazo/veterinaria , Proteínas Gestacionales/metabolismo , Animales , Bovinos , Enfermedades de los Bovinos/parasitología , Coccidiosis/metabolismo , Coccidiosis/parasitología , Femenino , Embarazo , Complicaciones Parasitarias del Embarazo/metabolismo , Proteínas Gestacionales/química
4.
Acta Orthop Belg ; 82(2): 280-286, 2016 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-27682290

RESUMEN

The outcome of a metaphyseal ulnar shortening osteotomy, reported by Sennwald as an alternative option for ulnar abutment syndrome, was evaluated with -regard to the functional results, pain relief and incidence of complications. METHOD: We conducted a review of 12 patients ; measured parameters were DASH score, PRWE, VAS for pain, grip force and wrist motion. RESULTS: Only disability improved significantly with a DASH score of 42 (p = 0.03). The function and pain assessment scores did not improve in 3 patients. We found no correlation between change in functional scores and other measured variables. Six patients were reoperated for prominent screw heads or new-onset distal radioulnar joint osteoarthritis. CONCLUSION: All osteomies healed fast and uneventfully and disability improved in most patients but 25% did not improve and 50% were re-operated. Further investigation is needed to evaluate the value of the metaphyseal osteotomy as an alternative treatment for ulnar abutment syndrome.


Asunto(s)
Osteotomía/métodos , Cúbito/cirugía , Adulto , Femenino , Fuerza de la Mano , Humanos , Masculino , Persona de Mediana Edad , Osteotomía/efectos adversos , Dolor/prevención & control , Complicaciones Posoperatorias , Radiografía , Rango del Movimiento Articular , Reoperación , Síndrome , Cúbito/diagnóstico por imagen , Cúbito/patología , Muñeca/fisiología , Adulto Joven
5.
Mamm Genome ; 26(1-2): 33-42, 2015 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-25308000

RESUMEN

Sequences encoding DUF1220 protein domains show the most extreme human lineage-specific copy number increase of any coding region in the genome and have been linked to human brain evolution. In addition, DUF1220 copy number (dosage) has been implicated in influencing brain size within the human species, both in normal populations and in individuals associated with brain size pathologies (1q21-associated microcephaly and macrocephaly). More recently, increasing dosage of a subtype of DUF1220 has been linked with increasing severity of the primary symptoms of autism. Despite these intriguing associations, a function for these domains has not been described. As a first step in addressing this question, we have developed the first transgenic model of DUF1220 function by removing the single DUF1220 domain (the ancestral form) encoded in the mouse genome. In a hypothesis generating exercise, these mice were evaluated by 197 different phenotype measurements. While resulting DUF1220-minus (KO) mice show no obvious anatomical peculiarities, they exhibit a significantly reduced fecundity (χ(2) = 19.1, df = 2, p = 7.0 × 10(-5)). Further extensive phenotypic analyses suggest hyperactivity (p < 0.05) of DUF1220 mice and changes in gene expression levels of brain associated with distinct neurological functions and disease. Other changes that met statistical significance include an increase in plasma glucose concentration (as measured by area under the curve, AUC 0-30 and AUC 30-120) in male mutants, fasting glucose levels, reduce sodium levels in male mutants, increased levels of the liver functional indicator ALAT/GPT in males, levels of alkaline phosphatase (also an indicator of liver function), mean R and SR amplitude by electrocardiography, elevated IgG3 levels, a reduced ratio of CD4:CD8 cells, and a reduced frequency of T cells; though it should be noted that many of these differences are quite small and require further examination. The linking of DUF1220 loss to a hyperactive phenotype is consistent with separate findings in which DUF1220 over expression results in a down-regulation of mitochondrial function, and potentially suggests a role in developmental metabolism. Finally, the substantially reduced fecundity we observe associated with KO mice argues that the ancestral DUF1220 domain provides an important biological functionthat is critical to survivability and reproductive success.


Asunto(s)
Evolución Biológica , Encéfalo/crecimiento & desarrollo , Fertilidad/genética , Dosificación de Gen , Ratones Transgénicos/genética , Fenotipo , Animales , Área Bajo la Curva , Glucemia/metabolismo , Calorimetría Indirecta , Cartilla de ADN/genética , Perfilación de la Expresión Génica , Técnicas de Inactivación de Genes , Hipercinesia/genética , Hígado/metabolismo , Masculino , Ratones , Tamaño de los Órganos , Estructura Terciaria de Proteína
6.
J Dairy Sci ; 98(7): 4593-600, 2015 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-25981074

RESUMEN

Coagulase-negative staphylococci (CNS) are the most common bacteria involved in subclinical mastitis in dairy cows. Remarkably, CNS-infected dairy heifers produce more milk than uninfected heifers. Because the lactation hormone prolactin (PRL) is also involved in mammary gland immunity, we investigated the milk PRL response and the mammary quarter milk yield following experimental CNS challenge. Eight healthy Holstein-Friesian heifers in mid-lactation were experimentally infected using a split-udder design with 3 different CNS strains: one Staphylococcus fleurettii (from sawdust bedding) and 2 Staphylococcus chromogenes strains (one isolate from a teat apex, the other isolate from a chronic intramammary infection). Three mammary quarters per heifer were simultaneously inoculated with 1.0×10(6) cfu, whereas the remaining mammary quarter was infused with sterile phosphate-buffered saline, serving as a control. An existing radioimmunoassay was modified, validated, and used to measure PRL frozen-thawed milk at various time points until 78h after challenge. The mean milk PRL level tended to be higher in the CNS-challenged mammary quarters compared with the control mammary quarters (7.56 and 6.85ng/mL, respectively). The increase in PRL over time was significantly greater in the CNS-challenged mammary quarters than in the control mammary quarters. However, no difference was found in the PRL response when comparing each individual CNS strain with the control mammary quarters. The mean mammary quarter milk yield tended to be lower in the CNS-infected mammary quarters than in the control mammary quarters (1.73 and 1.98kg per milking, respectively). The greatest milk loss occurred in the mammary quarters challenged with the intramammary strain of S. chromogenes. Future observational studies are needed to elucidate the relation between PRL, the milk yield, and the inflammatory condition, or infection status, of the mammary gland.


Asunto(s)
Leche/metabolismo , Prolactina/metabolismo , Infecciones Estafilocócicas/veterinaria , Staphylococcus/crecimiento & desarrollo , Animales , Bovinos , Modelos Animales de Enfermedad , Femenino , Lactancia , Glándulas Mamarias Animales/microbiología , Mastitis Bovina/microbiología , Staphylococcus/aislamiento & purificación
7.
Reprod Domest Anim ; 49(2): 275-80, 2014 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24456132

RESUMEN

Gamma interferon (IFN-γ) production and cross-breed pregnancy have been attributed a role in protecting dairy cows infected with Neospora caninum against abortion. Plasma levels of pregnancy-associated glycoproteins-1 (PAG-1) are a marker of placental/foetal well-being and of PAG-2 is an abortion risk indicator in chronically N. caninum-infected animals. The present study examines, in cross-breed pregnancies, interactions between IFN-γ production and levels of PAG-1 and PAG-2 in non-aborting naturally Neospora-infected dairy cows. Data were obtained from 60 pregnant Holstein-Friesian cows: 44 Neospora-seropositive and 16 Neospora-seronegative; 12 became pregnant using Holstein-Friesian semen and 48 using Limousin semen. Blood samples were collected on Days 40, 90, 120, 150, 180 and 210 of gestation. Gamma interferon was only detected in the plasma of nine of the 44 Neospora-seropositive cows, all of them became pregnant using Limousin semen. Through GLM procedures, in cows inseminated with Limousin semen and Neospora-seropositive cows showing no IFN-γ production, PAG-1 concentrations were high and increased throughout gestation compared to the levels detected in cows inseminated with Holstein-Friesian semen and Neospora-seropositive cows producing IFN-γ, respectively. In Neospora-seronegative cows and in Neospora-seropositive cows showing no IFN-γ production, significantly increased PAG-2 concentrations were observed on gestation Day 120. Our findings indicate that IFN-γ production correlates negatively and the production of antibodies against N. caninum is uncorrelated with plasma PAG concentrations during gestation in Neospora-infected dairy cows. Accordingly, IFN-γ production could be linked to the transplacental migration of tachyzoites, which may cause a reduction in PAG levels.


Asunto(s)
Ácido Aspártico Endopeptidasas/metabolismo , Coccidiosis/veterinaria , Interferón gamma/metabolismo , Complicaciones Parasitarias del Embarazo/veterinaria , Proteínas Gestacionales/metabolismo , Animales , Ácido Aspártico Endopeptidasas/sangre , Ácido Aspártico Endopeptidasas/genética , Bovinos , Coccidiosis/metabolismo , Industria Lechera , Femenino , Regulación de la Expresión Génica/fisiología , Interferón gamma/genética , Neospora , Embarazo , Proteínas Gestacionales/sangre , Proteínas Gestacionales/genética
8.
J Anim Physiol Anim Nutr (Berl) ; 98(5): 845-52, 2014 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-25356485

RESUMEN

The aim of this study is to evaluate the efficacy of frozen Azawak colostrum supplementation on body weight (BW), average daily gain (ADG), reproductive parameters (mean age at first parturition, fertility, fecundity, prolificacy) and mortality rate among red kids. The study was conducted at the goat farm secondary centre of Maradi in Niger from September 2010 to September 2011. The control animals (n = 20) were left with their mother, while the treatment animals (n = 20) received in addition 50 ml/animal/day of bovine colostrum at birth and 15 ml/animal/day from d2 to d15. Weight was measured weekly from birth to d365. Mortalities were also recorded over the same period. For reproductive parameters, observations began at weaning (d197). Growth rate was higher (p < 0.001) in supplemented animal, and the treatment effects on ADG were observed up to 150 day after the end of supplementation. A similar long-lasting trend was also observed in relation to the mortality rate (25% for ColG vs. 55% for ConG; p = 0.05). The age at first kidding tended to be lower in the treated group (13.8 ± 0.7 vs. 14.1 ± 0.8 month; p < 0.1). In conclusion, mild bovine colostrum supplementation induces a long-lasting positive impact on growth rate and to a lower extent on reproduction parameters and survival rate.


Asunto(s)
Animales Recién Nacidos/fisiología , Animales Lactantes/fisiología , Calostro/metabolismo , Cabras/crecimiento & desarrollo , Fenómenos Fisiológicos Nutricionales de los Animales , Animales , Bovinos , Femenino , Longevidad , Niger , Estado Nutricional , Análisis de Supervivencia , Aumento de Peso
9.
Schweiz Arch Tierheilkd ; 156(12): 585-90, 2014 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-25497564

RESUMEN

The aim of this study was to compare two methods of early pregnancy diagnosis by determining pregnancy-associated glycoprotein (PAG) concentration in blood and PAG concentration in milk. Blood and milk samples were obtained on days 0 (AI day), 14, 21, 28, 35, 49, 63, 77, 91 and 105 of gestation from 60 lactating Holstein Frisian cows from one herd, carrying live fetuses. To determine PAG concentration a specific RIA system (RIA-706) was used. PAG concentration in blood and milk increased after 28 days of pregnancy, with blood concentrations being significantly higher than in milk. However, the accuracy of both tests at this time point was similar (sensitivity: 92 % in blood, 93 % in milk; specificity 53 % and 60 % respectively). None of the tests were able to detect open cows properly at this stage. On day 35 of gestation sensitivity (100 % for blood, 97 % for milk) and specificity (100 % for blood, 100 % for milk) were high enough to be used for reliable pregnancy diagnosis. The accuracy (sensitivity and specificity) for PAG concentrations in blood and milk for the rest of the study was 100 %. Our investigation shows that PAG determination in milk is a stress-free and non-invasive method for early pregnancy diagnosis in cattle.


Le but de cette étude était de comparer les concentrations de glycoprotéines associées à la gestation (PAG) dans le sang et le lait en vue d'un diagnostic de gestation précoce chez la vache. Des échantillons de sang et de lait ont été prélevés aux jours 0 (IA), 14, 21, 28, 35, 49, 63, 77, 91 et 105 de la gestation sur 60 vaches Holstein Frisonnes en lactation avec un fœtus vivant. On a utilisé pour mesurer la concentration de PAG une méthode RIA spécifique (RIA-706). Les concentrations de PAG dans le sang et le lait s'élevaient à partir du 28ème jour de gestation, les concentrations mesurées dans le sang étant nettement plus élevée que celles mesurées dans le lait. L'exactitude des deux tests était à ce moment-là similaires (sensitivité: 92 % dans le sang, 93 % dans le lait, spécificité: 53 % respectivement 60 %). Aucune des deux méthodes n'était, à ce stade, à même de distinguer une vache non portante avec justesse. Au 35ème jour de gestation, la sensitivité des deux méthodes (100 % dans le sang, 97 % dans le lait) et leur spécificité (100 % dans le sang, 100 % dans le lait) était assez élevées pour permettre un diagnostic de gestation sûr. L'exactitude (sensitivité et spécificité) des mesures de concentration de PAG dans le sang et le lait durant la suite de la gestation était de 100 %. Ces études montrent donc que la mesure de PAG dans le lait représente une méthode non-invasive et exempte de stress pour le diagnostic précoce de gestation chez la vache.


Asunto(s)
Ácido Aspártico Endopeptidasas/sangre , Leche/química , Proteínas Gestacionales/sangre , Pruebas de Embarazo/métodos , Preñez/sangre , Animales , Bovinos , Femenino , Embarazo , Pruebas de Embarazo/normas , Sensibilidad y Especificidad
10.
Reprod Domest Anim ; 48(4): 613-8, 2013 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-23228018

RESUMEN

The present study sought to determine: (i) the effects of Neospora caninum infection and twin pregnancy on plasma pregnancy-associated glycoprotein-2 (PAG-2) concentrations throughout pregnancy and (ii) whether plasma PAG-2 concentrations could predict abortion in N. caninum-infected cows. The study was performed on a commercial Holstein-Friesian dairy herd in northeastern Spain and the final data included those recorded in 53 non-aborting and 19 aborting animals. Blood samples were collected immediately before pregnancy diagnosis (on Days 40, 90, 120, 150, 180 and 210 post-insemination) in non-aborting cows or until the time of abortion detection in aborting cows. General lineal models (GLM) repeated measures anova revealed the different behaviour of PAG-1 and PAG-2, and significant effects of Neospora seropositivity, cool season and twin pregnancy on plasma PAG-2 concentrations throughout gestation (between-subject effects). In addition, based on the odds ratios, the likelihood of abortion increased in Neospora-seropositive cows (by a factor of 7.0) compared to seronegative animals and decreased in cows with a high plasma PAG-2 concentration (>4.5 ng/ml) on Day 120 of pregnancy (by a factor of 0.24), compared to the remaining cows. In conclusion, there is a relationship between plasma PAG-2 concentrations and the risk of abortion in Neospora-infected dairy cows. Thus, plasma PAG concentrations measured using anti-boPAG-2 antiserum on Day 120 of gestation could serve as an indicator of the abortion risk in N. caninum infected animals; values <4.5 ng/ml indicating a high risk of abortion in chronically infected animals.


Asunto(s)
Aborto Veterinario/sangre , Ácido Aspártico Endopeptidasas/sangre , Enfermedades de los Bovinos/parasitología , Coccidiosis/veterinaria , Neospora , Complicaciones Parasitarias del Embarazo/veterinaria , Aborto Veterinario/parasitología , Animales , Anticuerpos Antiprotozoarios/sangre , Ácido Aspártico Endopeptidasas/inmunología , Biomarcadores/sangre , Bovinos , Enfermedades de los Bovinos/sangre , Coccidiosis/sangre , Coccidiosis/complicaciones , Ensayo de Inmunoadsorción Enzimática/veterinaria , Femenino , Edad Gestacional , Sueros Inmunes/inmunología , Neospora/inmunología , Embarazo , Complicaciones Parasitarias del Embarazo/sangre , Radioinmunoensayo/veterinaria , España
11.
Microbiol Spectr ; 10(5): e0034922, 2022 10 26.
Artículo en Inglés | MEDLINE | ID: mdl-35972267

RESUMEN

Transposable elements (TEs) play a pivotal role in shaping diversity in eukaryotic genomes. The covered smut pathogen on barley, Ustilago hordei, encountered a recent genome expansion. Using long reads, we assembled genomes of 6 U. hordei strains and 3 sister species, to study this genome expansion. We found that larger genome sizes can mainly be attributed to a higher genome fraction of long terminal repeat retrotransposons (LTR-RTs). In the studied smut genomes, LTR-RTs fractions are the largest in U. hordei and are positively correlated with the mating-type locus sizes, which is up to ~560 kb in U. hordei. Furthermore, LTR-RTs were found to be associated with higher nucleotide substitution levels, as these occur in specific genome regions of smut species with a recent LTR-RT proliferation. Moreover, genes in genome regions with higher nucleotide substitution levels generally reside closer to LTR-RTs than other genome regions. Genome regions with many nucleotide substitutions encountered an especially high fraction of CG substitutions, which is not observed for LTR-RT sequences. The high nucleotide substitution levels particularly accelerate the evolution of secretome genes, as their more accessory nature results in substitutions that often lead to amino acid alterations. IMPORTANCE Genomic alteration can be generated through various means, in which transposable elements (TEs) can play a pivotal role. Their mobility causes mutagenesis in itself and can disrupt the function of the sequences they insert into. They also impact genome evolution as their repetitive nature facilitates nonhomologous recombination. Furthermore, TEs have been linked to specific epigenetic genome organizations. We report a recent TE proliferation in the genome of the barley covered smut fungus, Ustilago hordei. This proliferation is associated with a distinct nucleotide substitution regime that has a higher rate and a higher fraction of CG substitutions. This different regime shapes the evolution of genes in subjected genome regions. We hypothesize that TEs may influence the error-rate of DNA polymerase in a hitherto unknown fashion.


Asunto(s)
Nucleótidos , Retroelementos , Elementos Transponibles de ADN , Secretoma , Secuencias Repetidas Terminales , Aminoácidos , Proliferación Celular , Evolución Molecular
12.
Phys Rev Lett ; 106(11): 115002, 2011 Mar 18.
Artículo en Inglés | MEDLINE | ID: mdl-21469868

RESUMEN

We used microparticles under hypergravity conditions, induced by a centrifuge, in order to measure nonintrusively and spatially resolved the electric field strength as well as the particle charge in the collisional rf plasma sheath. The measured electric field strengths demonstrate good agreement with the literature, while the particle charge shows decreasing values towards the electrode. We demonstrate that it is indeed possible to measure these important quantities without changing or disturbing the plasma.

13.
Reprod Domest Anim ; 46(3): 463-70, 2011 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-20887399

RESUMEN

In vitro oocyte maturation can be influenced by oocyte source and maturation media composition. The aim of the present study was to compare the efficiency of a defined in vitro maturation medium (TCM199 supplemented with cysteamine and epidermal growth factor; Cys + EGF) with an undefined medium (TCM199 supplemented with follicle-stimulating hormone and follicular fluid; FSH + FF) for in vitro production (IVP) of ovine embryos, using oocytes obtained by laparoscopic ovum pick-up from FSH-stimulated [n=11; 158 cumulus-oocyte complexes (COCs)] and non-stimulated (n=16; 120 COCs) live ewes, as well as abattoir-derived oocytes (170 COCs). The produced blastocysts were vitrified and some of them were transferred to synchronized recipients. The best and the worst final yields of embryo IVP observed in this study were obtained using oocytes from FSH-stimulated ewes matured in FSH + FF (41.3%; 33/80) and in Cys + EGF (19.2%; 15/78) medium, respectively (p<0.01). No significant differences between both media were attained in the blastocyst development rate or in the final yield of embryo IVP using oocytes from non-stimulated ewes or abattoir-derived oocytes. The overall in vivo survival rate of the transferred vitrified blastocysts was 13.1% (8/61), without significant differences between oocyte sources or maturation media. In conclusion, under the experimental conditions of the present study, TCM199 supplemented with cysteamine and EGF is a convenient defined maturation medium for IVP of embryos from oocytes of live non-stimulated ewes or from oocytes of abattoir-derived ovaries. However, the best final yield of embryo IVP observed in this study was attained when oocytes came from FSH-stimulated donors and TCM199 was supplemented with FSH and follicular fluid.


Asunto(s)
Medios de Cultivo , Técnicas de Cultivo de Embriones/veterinaria , Oocitos/fisiología , Ovinos/embriología , Animales , Blastocisto/fisiología , Células Cultivadas , Criopreservación/veterinaria , Transferencia de Embrión , Desarrollo Embrionario , Femenino , Fertilización In Vitro/veterinaria , Hormona Folículo Estimulante , Líquido Folicular , Embarazo
14.
Reprod Domest Anim ; 46(4): 656-63, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21121967

RESUMEN

The objective of this study was to test the accuracy of genotype diagnosis after whole amplification of DNA extracted from biopsies obtained by trimming goat embryos and to evaluate the viability of biopsied embryos after vitrification/warming and transfer. Whole genome amplification (WGA) was performed using Multiple Displacement Amplification (MDA). Sex and prion protein (PRNP) genotypes were determined. Sex diagnosis was carried out by PCR amplification of ZFX/ZFY and Y chromosome-specific sequences. Prion protein genotype determination was performed on codons 142, 154, 211, 222 and 240. Embryos were collected at day 7 after oestrus and biopsied either immediately after collection (blastocysts and expanded blastocysts) or after 24 h of in vitro culture (compacted morulae). Biopsied embryos were frozen by vitrification. Vitrified whole embryos were kept as control. DNA of biopsies was extracted and amplified using MDA. Sex diagnosis was efficient for 97.4% of biopsies and PRNP genotyping was determined in 78.7% of biopsies. After embryo transfer, no significant difference was observed in kidding rate between biopsied and vitrified control embryos, whereas embryo survival rate was different between biopsied and whole vitrified embryos (p = 0.032). At birth, 100% of diagnosed sex and 98.2% of predetermined codons were correct. Offspring PRNP profiles were in agreement with parental genotype. Whole genome amplification with MDA kit coupled with sex diagnosis and PRNP genotype predetermination are very accurate techniques to genotype goat embryos before transfer. These novel results allow us to plan selection of scrapie-resistant genotypes and kid sex before transfer of cryopreserved embryo.


Asunto(s)
Blastocisto/fisiología , Genotipo , Priones/genética , Análisis para Determinación del Sexo/veterinaria , Animales , ADN/genética , Transferencia de Embrión , Femenino , Genoma , Cabras , Masculino , Embarazo , Índice de Embarazo
15.
Rev Sci Instrum ; 92(1): 013506, 2021 Jan 01.
Artículo en Inglés | MEDLINE | ID: mdl-33514186

RESUMEN

We present the design and commissioning of a resonant microwave cavity as a novel diagnostic for the study of ultracold plasmas. This diagnostic is based on the measurements of the shift in the resonance frequency of the cavity, induced by an ultracold plasma that is created from a laser-cooled gas inside. This method is simultaneously non-destructive, very fast (nanosecond temporal resolution), highly sensitive, and applicable to all ultracold plasmas. To create an ultracold plasma, we implement a compact magneto-optical trap based on a diffraction grating chip inside a 5 GHz resonant microwave cavity. We are able to laser cool and trap (7.25 ± 0.03) × 107 rubidium atoms inside the cavity, which are turned into an ultracold plasma by two-step pulsed (nanosecond or femtosecond) photo-ionization. We present a detailed characterization of the cavity, and we demonstrate how it can be used as a fast and sensitive probe to monitor the evolution of ultracold plasmas non-destructively. The temporal resolution of the diagnostic is determined by measuring the delayed frequency shift following femtosecond photo-ionization. We find a response time of 18 ± 2 ns, which agrees well with the value determined from the cavity quality factor and resonance frequency.

16.
Rev Sci Instrum ; 92(9): 093504, 2021 Sep 01.
Artículo en Inglés | MEDLINE | ID: mdl-34598523

RESUMEN

Low-temperature plasmas are widely studied in laboratory environments and form the backbone of many industrial processes. Highly energized electrons enable processes such as ionization, dissociation, and plasma chemical reactions, while the heavy species, such as neutral gas atoms and molecules, remain near room temperature. Hence, understanding the electron dynamics is crucial to the control and optimization of plasmas and their applications. In this contribution, we investigated the impact of electron density profile correction on microwave cavity resonance spectroscopy (MCRS) as a diagnostic tool for low-pressure discharges. Following standard practice, we first obtained a volume-averaged electron density by assuming a uniform plasma in the interpretation of the MCRS diagnostic technique. Second, we compare the experiments with a numerical model solved using PLASIMO software to evaluate the predictive capabilities. Third, we obtained profile-corrected electron densities by means of incorporating the numerically obtained distribution of the electron density and the numerical solution for the resonant microwave electric field in the interpretation of the experimental data using MCRS. Although the volume-averaged data agree closely with the electron density found from the numerical model, it is shown that implementing the spatial distribution of the electron density and the microwave electric field leads to a significant correction to the experimental data. The developed strategy could easily be implemented in other situations deploying MCRS as a non-invasive technique for measuring the electron density.

17.
Rev Sci Instrum ; 92(6): 063518, 2021 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-34243512

RESUMEN

A simple and selective new technique for atomic hydrogen flux measurements in a hydrogen plasma environment is introduced and demonstrated in this work. This technique works by measuring the etching rate of an amorphous carbon film and translating this to an incoming hydrogen radical flux through a well-defined carbon etch yield per radical. Ions present in the plasma environment have a much higher etch yield than radicals do. For that reason, suppression of the ion flux toward the carbon film is crucial to ensure that the observed carbon etch rate is dominated by atomic hydrogen etching. It is demonstrated that this can be achieved using a simple cylindrical pipe (hereinafter "chimney") in which a bend is introduced to enforce ion-wall collisions, neutralizing the ions. The chimney is made out of Macor, a material with low catalytic surface activity, to preserve the incoming atomic hydrogen flux while effectively suppressing ions. Ultimately, the etching sensor is deployed in a radio frequency inductively coupled hydrogen plasma operated at low pressure (1-10 Pa). Atomic hydrogen fluxes are measured and compared with heat flux sensor and vacuum ultraviolet absorption spectroscopy measurements in the same setup. All sensors agreed within a factor 4 in the atomic hydrogen flux range 1019 to 1021 m-2 s-1.

18.
Nat Commun ; 12(1): 4692, 2021 Aug 03.
Artículo en Inglés | MEDLINE | ID: mdl-34344881

RESUMEN

The plasma-induced charge of non-spherical microparticles is a crucial parameter in complex plasma physics, aerosol science and astrophysics. Yet, the literature describes this charge by two competing models, neither of which has been experimentally verified or refuted. Here we offer experimental proof that the charge on a two-particle cluster (doublet) in the spatial afterglow of a low-pressure plasma equals the charge that would be obtained by the smallest enclosing sphere and that it should therefore not be based on its geometrical capacitance but rather on the capacitance of its smallest enclosing sphere. To support this conclusion, the size, mass and charge of single particles (singlets) and doublets are measured with high precision. The measured ratio between the plasma-afterglow-induced charges on doublets and singlets is compared to both models and shows perfect agreement with the predicted ratio using the capacitance of the smallest enclosing sphere, while being significantly dissimilar to the predicted ratio based on the particle's geometrical capacitance.

19.
Reprod Domest Anim ; 45(6): e231-8, 2010 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-19735438

RESUMEN

Thirteen female Bedouin goats living in arid land of Algeria Sahara desert were used in this study. These goats were pregnant but they sustained an abortion because of unidentified causes. None of the goats showed any signs of general disease. Plasma concentrations of caprine pregnancy-associated glycoproteins (cPAGs) and progesterone (P4) were determined during pregnancy using radioimmunoassay. The cPAGs concentration was undetectable (<0.8 ng/ml) throughout the first 2 weeks of gestation. From week 3 after mating, cPAGs concentration was detectable with significant individual variations (p < 0.05) reaching a maximum secretion (436.1 ng/ml). Throughout gestation, cPAGs concentration remained relatively constant but decreased few days before abortion, on an average of 9.2 ± 1.2 days (n = 11), except for two females where the concentrations decreased later (1-2 days before abortion). One or two peaks of cPAGs concentrations (in 4/13 and in 9/13 females, respectively) have been measured few weeks before abortion (77-124 days after mating), when a decline of cPAGs was detected. The P4 concentration increased after mating, and was high from the first week till the end of pregnancy. The P4 concentration (9.1 ± 0.9 ng/ml) decreased rapidly (<0.5 ng/ml) after 4 ± 0.7 days (n = 6) or 9.4 ± 1.6 days (n = 7) before abortion. A positive relationship (p < 0.01) was found between P4 and cPAGs concentrations during gestation. Results indicate that cPAGs and P4 measurements can be used for monitoring gestation and for abortion prediction.


Asunto(s)
Aborto Veterinario/sangre , Glicoproteínas/sangre , Enfermedades de las Cabras/sangre , Progesterona/sangre , Aborto Veterinario/epidemiología , Argelia/epidemiología , Animales , Femenino , Cabras , Embarazo
20.
Rev Sci Instrum ; 91(6): 063503, 2020 Jun 01.
Artículo en Inglés | MEDLINE | ID: mdl-32611041

RESUMEN

For the microwave cavity resonance spectroscopy based non-destructive beam monitor for ionizing radiation, an addition-which adapts the approach to conditions where only little ionization takes place due to, e.g., small ionization cross sections, low gas pressures, and low photon fluxes-is presented and demonstrated. In this experiment, a magnetic field with a strength of 57 ± 1 mT was used to extend the lifetime of the afterglow of an extreme ultraviolet-induced plasma by a factor of ∼5. Magnetic trapping is expected to be most successful in preventing the decay of ephemeral free electrons created by low-energy photons. Good agreement has been found between the experimental results and the decay rates calculated based on the ambipolar and classical collision diffusion models.

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