1,690 research outputs found

    Expected Response To Early-generation Selection For Yield And Tuber Appearance Traits In Potatoes

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    The aim of this study was to evaluate the performance of potato clonal families, and to estimate genetic variance, heritability and the expected response to selection of tuber yield and appearance traits in early generations. Twelve potato families were obtained from crosses between two groups of randomly-selected genotypes, including Eliza, C1730-7-94, and C1742-8-95 in group 1; and Shepody, Asterix, Caesar, and White Lady in group 2. The crosses were made in factorial design (3 genotypes x 4 genotypes), and each family consisted of 75 genotypes. Experiments were conducted in the fall of 2010, with a seedling generation under greenhouse conditions, and in the fall of 2011, with a clonal generation under field condition. High heritability estimates suggest that mild to moderate selection can be applied in the seedling generation to eye depth, eyebrow prominence, tuber curvature, flattening and shape uniformity. The C1742-8-95/White Lady stood out as a superior cross, as did all other crosses with White Lady, regarding tuber appearance and yield traits.3752849285

    Novel pharmacological actions of Trequinsin Hydrochloride improve human sperm cell motility and function

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    Background and purposeAsthenozoospermia is a leading cause of male infertility, but the development of pharmaceuticals to improve sperm motility has been hindered by the lack of effective screening platforms and knowledge of suitable molecular targets. We have demonstrated that a high throughput screening (HTS) strategy in conjunction with established in vitro tests can identify and characterise the action of compounds that improve sperm motility. The study aimed to apply HTS to identify new compounds from a novel small molecule library that increase intracellular calcium, [Ca2+]I, promote human sperm cell motility and systemically determine the mechanism of action. Experimental approach A validated HTS fluorometric [Ca2+]i assay was used to screen an in-house library of compounds. Trequinsin hydrochloride (a phosphodiesterase 3 inhibitor) was selected for detailed molecular (plate reader assays, electrophysiology and cyclic nucleotide measurement) and functional (motility and acrosome reaction) testing in sperm from healthy volunteer donors and, where possible, patients.   Key resultsThe fluorometric analysis identified Trequinsin as an efficacious agonist of [Ca2+]i, although less potent than progesterone (P4). Functionally, Trequinsin significantly increased cell hyperactivation and penetration into viscous medium in all donor sperm samples and cell hyperactivation in 22/25 (88%) patient sperm samples. The Trequinsin-induced [Ca2+]i response was cross-desensitised consistently by prostaglandin E1 but not with P4. Whole-cell patch clamp electrophysiology confirmed that Trequinsin activates CatSper and partially inhibits potassium channel activity. Trequinsin also increases intracellular cGMP.   Conclusion and Implications Trequinsin exhibits a novel pharmacological profile in human sperm and may be a suitable lead compound for the development of new pharmaceuticals to improve patient sperm function and fertilisation potential

    Complex CatSper-dependent and independent [Ca2<sup>+</sup>]i signalling in human spermatozoa induced by follicular fluid

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    STUDY QUESTION: Does progesterone in human follicular fluid (hFF) activate CatSper and do other components of hFF modulate this effect and/or contribute separately to hFF-induced Ca2+ signaling?SUMMARY ANSWER: hFF potently stimulates CatSper and increases [Ca2+]i, primarily due to high concentrations of progesterone, however,other components of hFF also contribute to [Ca2+]i signaling, including modulation of CatSper channel activity and inhibition of [Ca2+]i oscillations.WHAT IS KNOWN ALREADY: CatSper, the principal Ca2+ channel in spermatozoa, is progesterone-sensitive and essential for fertility. Both hFF and progesterone, which is present in hFF, influence sperm function and increase their [Ca2+]i.STUDY DESIGN, SIZE, DURATION: This basic medical research study used semen samples from &gt;40 donors and hFF from &gt;50 patients who were undergoing surgical oocyte retrieval for IVF/ICSI.PARTICIPANTS/MATERIALS, SETTING, METHODS: Semen donors and patients were recruited in accordance with local ethics approval (13/ES/0091) from the East of Scotland Research Ethics Service REC1. Activities of CatSper and KSper were assessed by patch clamp electrophysiology. Sperm [Ca2+]i responses were examined in sperm populations and single cells. Computer-assisted sperm analysis (CASA) parameters and penetration into viscous media were used to assess functional effects.MAIN RESULTS AND THE ROLE OF CHANCE: hFF and progesterone significantly potentiated CatSper currents. Under quasiphysiologicalconditions, hFF (up to 50%) failed to alter membrane K+ conductance or current reversal potential. hFF and progesterone (at an equivalent concentration) stimulated similar biphasic [Ca2+]i signals both in sperm populations and single cells. At a high hFF concentration (10%), the sustained (plateau) component of the [Ca2+]i signal was consistently greater than that induced by progesterone alone. In single cell recordings, 1% hFF-induced [Ca2+]i oscillations similarly to progesterone but with 10% hFF generation of [Ca2+]i oscillations was suppressed. After treatment to ‘strip’ lipid-derived mediators, hFF failed to significantly stimulate CatSper currents but induced small [Ca2+]i responsesthat were greater than those induced by the equivalent concentration of progesterone after stripping. Similar [Ca2+]i responses were observed when sperm pretreated with 3 μM progesterone (to desensitize progesterone responses) were stimulated with hFF or stripped hFF. hFF stimulated viscous media penetration and was more effective than the equivalent does of progesterone.LARGE SCALE DATA: N/A.LIMITATIONS, REASONS FOR CAUTION: This was an in vitro study. Caution must be taken when extrapolating these results in vivo.WIDER IMPLICATIONS OF THE FINDINGS: This study directly demonstrates that hFF activates CatSper and establishes that the biologically important effects of hFF reflect, at least in part, action on this channel, primarily via progesterone. However, these experiments also demonstrate that other components of hFF both contribute to the [Ca2+]i signal and modulate the activation of CatSper. Simple in vitro experiments performed out of the context of the complex in vivo environment need to be interpreted with caution

    Single-cell analysis of [Ca<sup>2+</sup>]i signalling in sub-fertile men:characteristics and relation to fertilization outcome

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    STUDY QUESTIONWhat are the characteristics of progesterone-induced (CatSper-mediated) single cell [Ca2+]i signals in spermatozoa from sub-fertile men and how do they relate to fertilizing ability?SUMMARY ANSWERSingle cell analysis of progesterone-induced (CatSper-mediated) [Ca2+]i showed that reduced progesterone-sensitivity is a common feature of sperm from sub-fertile patients and is correlated with fertilization rate.WHAT IS KNOWN ALREADYStimulation with progesterone is a widely used method for assessing [Ca2+]i mobilization by activation of CatSper in human spermatozoa. Although data are limited, sperm population studies have indicated an association of poor [Ca2+]i response to progesterone with reduced fertilization ability.STUDY DESIGN, SIZE, DURATIONThis was a cohort study using semen samples from 21 donors and 101 patients attending the assisted conception unit at Ninewells Hospital Dundee who were undergoing ART treatment. Patients were recruited from January 2016 to June 2017.PARTICIPANTS/MATERIALS, SETTING, METHODSSemen donors and patients were recruited in accordance with local ethics approval (13/ES/0091) from the East of Scotland Research Ethics Service (EoSRES) REC1. [Ca2+]i responses were examined by single cell imaging and motility parameters assessed by computer-assisted sperm analysis (CASA).MAIN RESULTS AND THE ROLE OF CHANCEFor analysis, patient samples were divided into three groups IVF(+ve) (successful fertilization; 62 samples), IVF-FF (failed fertilization; eight samples) and ICSI (21 samples). A further 10 IVF samples showed large, spontaneous [Ca2+]i oscillations and responses to progesterone could not be analysed. All patient samples loaded with the [Ca2+]i-indicator fluo4 responded to progesterone stimulation with a biphasic increase in fluorescence (transient followed by plateau) which resembled that seen in progesterone-stimulated donor samples. The mean normalized response (progesterone-induced increase in fluorescence normalized to resting level) was significantly smaller in IVF-FF and ICSI patient groups than in donors. All samples were further analysed by plotting, for each cell, the relationship between resting fluorescence intensity and the progesterone-induced fluorescence increment. In donor samples these plots overlaid closely and had a gradient of ≈ 2 and plots for most IVF(+ve) samples closely resembled the donor distribution. However, in a subset (≈ 10%) of IVF(+ve) samples, 3/8 IVF-FF samples and one-third of ICSI samples the gradient of the plot was significantly lower, indicating that the response to progesterone of the cells in these samples was abnormally small. Examination of the relationship between gradient (regression coefficient of the plot) in IVF samples and fertilization rate showed a positive correlation. In IVF-FF and ICSI groups, the proportion of cells in which a response to progesterone could be detected was significantly lower than in donors and IVF (+ve) patients. Approximately 20% of cells in donor, IVF(+ve) and ICSI samples generated [Ca2+]i oscillations when challenged with progesterone but in IVF-FF samples only ≈ 10% of cells generated oscillations and there was a significantly greater proportion of samples where no oscillations were observed. Levels of hyperactivated motility were lower in IVF(+ve) and IVF-FF groups compared to controls, IVF-FF also having lower levels than IVF(+ve).LIMITATIONS, REASONS FOR CAUTIONThis is an in vitro study and caution must be taken when extrapolating these results in vivo.WIDER IMPLICATIONS OF THE FINDINGSThis study reveals important details of impaired [Ca2+]i signalling in sperm from sub-fertile men that cannot be detected in population studies

    Anomalous diffusion and anisotropic nonlinear Fokker-Planck equation

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    We analyse a bidimensional nonlinear Fokker-Planck equation by considering an anisotropic case, whose diffusion coefficients are DxxθD_x \propto |x|^{-\theta} and DyyγD_y \propto |y|^{-\gamma} with θ,γR\theta, \gamma \in {\cal{R}}. In this context, we also investigate two situations with the drift force F(r,t)=(kxx,kyy)\vec{F}(\vec{r},t)=(-k_{x}x, -k_y y). The first one is characterized by kx/ky=(2+γ)/(2+θ)k_x/k_y=(2+\gamma)/(2+\theta) and the second is given by kx=kk_{x}=k and ky=0k_{y}=0. In these cases, we can verify an anomalous behavior induced in different directions by the drift force applied. The found results are exact and exhibit, in terms of the qq-exponentials, functions which emerge from the Tsallis formalism. The generalization for the DD-dimensional case is discussed.Comment: 6 pages, tex fil

    Postnatal development of rats exposed to fluoxetine or venlafaxine during the third week of pregnancy

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    The aim of the present study was to compare the toxic effects of fluoxetine (F) (8 and 16 mg/kg) and venlafaxine (V) (40 and 80 mg/kg) administered during the third week of pregnancy on early development of rats. Both antidepressants were administered by gavage on pregnancy days 15 to 20 to groups of 10 to 12 animals each. Duration of gestation, food and water consumption, number of live pups and birth weight were recorded. Litters were culled to six pups at birth (day 1) and followed for growth until weaning (day 25). On day 60, a male and a female from each litter were injected with the 5-HT1 agonist, 5-methoxy-N,N-dimethyltryptamine (6 mg/kg, ip) and the serotonergic syndrome was graded. Fluoxetine but not venlafaxine reduced the duration of pregnancy when compared to the control (C) group (F = 21.1 days and C = 21.6 days, mean, P<0.02; maximum = 22 days and minimum = 21 days in both groups). The highest doses of both fluoxetine, 16 mg/kg (F16), and venlafaxine, 80 mg/kg (V80), reduced the food intake of pregnant rats, resulting in different rates of body weight gain during treatment (from pregnancy day 15 to day 20): F16 = 29.0 g, V80 = 28.7 g vs C = 39.5 g (median). Birth weight was influenced by treatment and sex (P<0.05; two-way ANOVA). Both doses of fluoxetine or venlafaxine reduced the body weight of litters; however, the body weight of litters from treated dams was equal to the weight of control litters by the time of weaning. At weaning there was no significant difference in weight between sexes. There was no difference among groups in number of live pups at birth, stillbirths, mortality during the lactation period or in the manifestation of serotonergic syndrome in adult rats. The occurrence of low birth weight among pups born to dams which did not show reduced food ingestion or reduction of body weight gain during treatment with lower doses of fluoxetine or venlafaxine suggests that these drugs may have a deleterious effect on prenatal development when administered during pregnancy. In addition, fluoxetine slightly but significantly affected the duration of pregnancy (about half a day), an effect not observed in the venlafaxine-treated groups.Universidade Federal FluminenseUniversidade Federal de São Paulo (UNIFESP)UNIFESPSciEL

    Determination of low molecular weight volatiles in Ficus carica using HS-SPME and GC/FID

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    Ficus carica L. is one of the earliest cultivated fruit trees, having an important consumption in Mediterranean countries. In this work, the volatile compound profiles of two characteristic Portuguese white varieties (‘‘Pingo de Mel” and ‘‘Branca Tradicional”) was determined by HS-SPME and GC/FID. Leaves, pulps and peels, submitted to freezing and lyophilisation treatments, were analysed. The two varieties presented a similar profile composed of eight volatile compounds: acetaldehyde, ethyl acetate, methanol, ethanol, hexanal, limonene, (E)-2-hexenal and octanal. The total volatile content was different among the vegetal materials, following the order leaves > peels > pulps. Methanol and ethanol are the major compounds in all samples. The developed procedure revealed to be rapid, sensitive, reproducible and accurate. The detection limit values were low, and the method precise. The recovery values for acetaldehyde, ethyl acetate, methanol and ethanol were generally high, suggesting that it will be most suitable for compounds with low molecular weight. Due to its rapidity and low cost, this technique can be useful in the quality control of fig fruit and leaves

    Tripping over emerging pathogens around the world: A phylogeographical approach for determining the epidemiology of Porcine circovirus-2 (PCV-2), considering global trading

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    AbstractPorcine circovirus-2 (PCV-2) is an emerging virus associated with a number of different syndromes in pigs known as Porcine Circovirus Associated Diseases (PCVAD). Since its identification and characterization in the early 1990s, PCV-2 has achieved a worldwide distribution, becoming endemic in most pig-producing countries, and is currently considered as the main cause of losses on pig farms. In this study, we analyzed the main routes of the spread of PCV-2 between pig-producing countries using phylogenetic and phylogeographical approaches. A search for PCV-2 genome sequences in GenBank was performed, and the 420 PCV-2 sequences obtained were grouped into haplotypes (group of sequences that showed 100% identity), based on the infinite sites model of genome evolution. A phylogenetic hypothesis was inferred by Bayesian Inference for the classification of viral strains and a haplotype network was constructed by Median Joining to predict the geographical distribution of and genealogical relationships between haplotypes. In order to establish an epidemiological and economic context in these analyses, we considered all information about PCV-2 sequences available in GenBank, including papers published on viral isolation, and live pig trading statistics available on the UN Comtrade database (http://comtrade.un.org/). In these analyses, we identified a strong correlation between the means of PCV-2 dispersal predicted by the haplotype network and the statistics on the international trading of live pigs. This correlation provides a new perspective on the epidemiology of PCV-2, highlighting the importance of the movement of animals around the world in the emergence of new pathogens, and showing the need for effective sanitary barriers when trading live animals

    Spatial distribution of the most abundant sea urchin populations on the southeast coast of São Paulo (Brazil)

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    Se investigó la variación espacial en la abundancia de Echinometra lucunter, Lytechinus variegatus y Arbacia lixula con respecto a cuatro escalas espaciales (105, 104, 103 y 102 m) y dos profundidades (0–3 y 3–8 m), a lo largo de 100 km del litoral de São Paulo (agosto a octubre de 1996). Echinometra lucunter fue la especie más abundante, prefiriendo los fondos más someros bajo el efecto del oleaje. Se identificó un gradiente en su abundancia, aumentando en dirección del suroeste al noreste. Arbacia lixula mostró el mismo gradiente, pero con una mayor abundancia en el sector más profundo. Ambas especies mostraron diferencias significativas a una escala espacial de centenares de metros y E. lucunter también a una escala de kilómetros. Lytechinus variegatus mostró un patrón en mosaico, siendo abundante en ciertos sitios. Las variaciones en la calidad del agua y en la heterogeneidad natural del hábitat podrían explicar de forma conjunta la distribución espacial de estas poblaciones.The spatial variability of Echinometra lucunter, Lytechinus variegatus and Arbacia lixula was studied in relation to four spatial scales (105, 104, 103 and 102 m) and two depths (0–3 and 3–8 m), along 100 km of the São Paulo coastline (August to October 1996). Echinometra lucunter was the most abundant species, with preference for shallow substrates subject to wave action. An abundance gradient was identified, increasing from southwest to northeast. Arbacia lixula showed the same pattern of spatial variation, but was more abundant in the deeper areas. Both species showed significant differences on a spatial scale of hundreds of meters, and E. lucunter also on a scale of kilometers. Lytechinus variegates presented a patchy distribution, being more abundant at certain sites. Variations in the water quality and natural heterogeneity of the habitat may explain the spatial distribution of these populations.El presente trabajo ha sido realizado gracias a dos becas INTERCAMPUS (1996–1997) concedidas por el Gobierno Español (Ministerio de Asuntos Exteriores) a PSJ para su estancia en el Laboratorio de Ecotoxicología (Brasil)

    Laparoscopic antireflux surgery in patients with extra esophageal symptoms related to asthma

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    Asthma, laryngitis and chronic cough are atypical symptoms of the gastroesophageal reflux disease. To analyze the efficacy of laparoscopic surgery in the remission of extra-esophageal symptoms in patients with gastroesophageal reflux, related to asthma. Were reviewed the medical records of 400 patients with gastroesophageal reflux disease submitted to laparoscopic Nissen fundoplication from 1994 to 2006, and identified 30 patients with extra-esophageal symptoms related to asthma. The variables considered were: gender, age, gastroesophageal symptoms (heartburn, acid reflux and dysphagia), time of reflux disease, treatment with proton pump inhibitor, use of specific medications, treatment and evolution, number of attacks and degree of esophagitis. Data were subjected to statistical analysis, comparing the pre- and post-surgical findings. The comparative analysis before surgery (T1) and six months after surgery (T2) showed a significant reduction on heartburn and reflux symptoms. Apart from that, there was a significant difference between the patients with daily crises of asthma (T1 versus T2, 45.83% to 16.67%, p=0.0002) and continuous crises (T1, 41.67% versus T2, 8.33%, p=0.0002). Laparoscopic Nissen fundoplication was effective in improving symptoms that are typical of reflux disease and clinical manifestations of asthma.Asthma, laryngitis and chronic cough are atypical symptoms of the gastroesophageal reflux disease. To analyze the efficacy of laparoscopic surgery in the remission of extra-esophageal symptoms in patients with gastroesophageal reflux, related to asthma. W2729295sem informaçãosem informaçã
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