Provision of choline chloride to the bovine preimplantation embryo alters postnatal body size and DNA methylation.

Choline is a vital micronutrient that can be utilized in the formation of betaine and multiple phospholipids. In this study, we aimed to confirm, and expand on previous findings, how choline impacts embryos from the first 7 days of development to affect postnatal phenotype. Bos indicus embryos were cultured in a choline-free medium (termed vehicle) or medium supplemented with 1.8 mM choline Blastocyst-stage embryos were transferred into crossbred recipients. Once born, calves were evaluated at birth, 94 d, 178 d and at weaning (average age = 239 d). Following weaning, all calves were enrolled into a feed efficiency trial before being separated by sex, with males being slaughtered at approximately 580 d of age and females followed until their first pregnancy check. Results confirm that exposure of 1.8 mM choline chloride during the first 7 d of development alters postnatal characteristics of the resultant calves. Calves of both sexes from choline-treated embryos were consistently heavier through weaning and males had heavier testes at 3 mo of age. There were sex-dependent alterations in DNA methylation in whole blood caused by choline treatment. After weaning, feed efficiency was affected by an interaction with sex, with choline calves being more efficient for females and less efficient for males. Calves from choline-treated embryos were heavier, or tended to be heavier, than calves from vehicle embryos at all observations after weaning. Carcass weight was heavier for choline calves and the cross-sectional area of the Longissumus thoracis muscle was increased by choline. Few females became pregnant during the experiment although numerically more choline females were pregnant than vehicle females. Results confirm that exposure of the preimplantation embryo to 1.8 mM choline can alter phenotypes of the resultant calves through the first 19 months after birth.

Cycloaddition between nitrogen-doped graphene (6π-component) and benzene (4π-component): a theoretical approach using density functional theory with vdW-DF correction.

The interaction between nitrogen-doped graphene defects (N3V1 and N4V2 pyridinic, and N3V1 and N3V3 pyrrolic) and benzene have been investigated by applying density functional theory (DFT), together with the vdW-DF correction. We discovered that only the N3V3 pyrrolic defect is a reactive site (6π-component), forming a cycloadduct with benzene (4π-component) that has energy barriers below 154.38 kJ mol-1 (1.60 eV). The conduction and valence bands (HOMO and LUMO) for N3V3 form a degenerate pair of orbitals at the gamma point, with the same ionization potential (IP) and electron affinity (EA). Likewise, inspection of the orbital symmetries for both systems confirms that these must undergo concerted reactions based on the Woodward and Hoffmann principles of orbital symmetry, with the appropriate orbital occupancies. This is the first time that substitutionally doped graphene has been demonstrated to participate as a 6π-component for cycloaddition reactions with benzene.

Actions of putative embryokines on development of the preimplantation bovine embryo to the blastocyst stage.

Once it enters the uterus at d 4 to 5 after ovulation, the preimplantation bovine embryo is controlled in its development by regulatory signaling molecules from the mother called embryokines. Here, several cell-signaling molecules whose genes are expressed in the endometrium during d 5 to 7 after estrus were tested for the ability to affect the competence of the embryo for further development and the characteristics of the resultant blastocysts. Molecules tested were C-natriuretic peptide (CNP), IL-8, bovine morphogenetic protein 4 (BMP-4), IL-6, and leukemia inhibitory factor (LIF). None of the cell-signaling molecules tested improved the competence of the embryo to become a blastocyst; in fact, BMP-4 decreased development. All molecules modified attributes of the blastocyst formed in culture. In particular, CNP increased the number of cells in the ICM, whereas IL-8 decreased inner cell mass cell numbers and tended to increase the proportion of blastocysts that were hatching or hatched. In addition, BMP-4 decreased the proportion of blastocysts that were hatching. Interleukin-6 and, to a lesser extent, LIF activated the Janus kinase (JAK)/signal transducer and activator of transcription 3 (STAT3) signaling pathway in the inner cell mass, and LIF increased the percent of cells in the blastocyst that were positive for both NANOG and phosphorylated (activated) STAT3. In conclusion, our results indicate that CNP, IL-8, IL-6, LIF, and BMP-4 can modify embryonic development of the cow in a manner that affects characteristics of the resultant blastocyst. Further research is required to understand how these changes in characteristics of the blastocyst would affect competence of the embryo to establish and maintain pregnancy.

Effects of choline on the phenotype of the cultured bovine preimplantation embryo.

Choline is a precursor of acetylcholine, phosphatidylcholine, and the methyl-donor betaine. Reports indicate that supplementation with rumen-protected choline improves postpartum reproductive function of dairy cows. The objective was to determine whether addition of choline to culture medium of in vitro-produced embryos alters the phenotype of the resultant blastocysts. Treatments were choline chloride (ChCl; 0.004, 1.3, 1.8, and 6.37 mM) and phosphatidylcholine (1.3 mM). Treatment with 0.004 mM ChCl improved development to the blastocyst stage, increased blastocyst cell number, and increased the percentage of blastocysts that were hatching or hatched. Development was not affected by higher concentrations of ChCl but was reduced by 1.3 mM phosphatidylcholine. Treatment of embryos with 1.3 mM ChCl (but not other concentrations) increased expression in blastocysts of 11 of 165 genes examined (AMOT, NANOG, HDAC8, HNF4A, STAT1, MBNL3, SOX2, STAT3, KDM2B, SAV1, and GPAM) and decreased expression of one gene (ASS1). Treatment with 1.3 mM ChCl decreased global DNA methylation at d 3.5 of development and increased DNA methylation at d 7.5 in blastocysts. Treatment with 1.8 mM ChCl also increased methylation in blastocysts. In conclusion, addition of choline to the culture medium alters the phenotype of preimplantation bovine embryos produced in vitro. Choline chloride can act in a concentration-dependent manner to alter development, expression of specific genes, and DNA methylation.

Effects of intrauterine infusion of seminal plasma at artificial insemination on fertility of lactating Holstein cows.

An inflammatory response is induced in the reproductive tract by deposition of semen during natural mating. This response might facilitate establishment and maintenance of pregnancy and alter the phenotype of the offspring by modifying the microenvironment of the reproductive tract. Here, we hypothesized that intrauterine infusion of 0.5 mL of seminal plasma at the time of artificial insemination (AI) in first-service lactating Holstein cows will improve pregnancy success after insemination. Cows were inseminated (511 primiparous cows inseminated with X-sorted semen, 554 multiparous cows inseminated with X-sorted semen, and 627 multiparous cows inseminated with conventional semen) using the Double-Ovsynch protocol. Cows were randomly assigned to receive intrauterine infusion of either 0.5 mL of seminal plasma or saline immediately after AI. There was no overall effect of seminal plasma infusion on the percentage of inseminated cows diagnosed pregnant at d 32 or 60 after AI, pregnancy loss, or percent of inseminated cows calving. If cows were inseminated with conventional semen, seminal plasma reduced pregnancies at d 32 and tended to reduce calvings. There was no effect of seminal plasma if cows were inseminated with X-sorted semen. Seminal plasma infusion increased the birth weight of heifer calves born using X-sorted semen but not conventional semen. These results do not support a beneficial effect of seminal plasma on pregnancy success after AI, but exposure to seminal plasma may program fetal development to affect phenotype at birth.

Interactions of human chorionic gonadotropin with genotype and parity on fertility responses of lactating dairy cows.

Fertility-promoting effects of treatment of lactating dairy cattle with human chorionic gonadotropin (hCG) after artificial insemination (AI) have been variable. Here, we tested whether fertility response to hCG in lactating Holstein cows interacts with genotype and parity. Primiparous (n = 538) and multiparous (n = 613) cows were treated with hCG (3,300 IU) or vehicle 5 d after AI. Pregnancy was diagnosed on d 32 and 60 after AI. A subset of cows (n = 593-701) was genotyped for 4 single nucleotide polymorphisms (SNP) previously associated with fertility. Treatment with hCG increased progesterone concentration on d 12 after AI regardless of genotype or parity. Pregnancy per AI was improved by hCG in primiparous cows but not in multiparous cows. Moreover, hCG treatment interacted with a SNP in coenzyme Q9 (COQ9) to affect fertility. Fertility of cows treated with vehicle was greatest for the AA allele, whereas fertility was lowest for the same genotype among cows treated with hCG. Pregnancy per AI was also affected by genotype for heat shock protein A1-like (HSPA1L) and progesterone receptor (PGR), but no interactions were observed with treatment. Genotype for a SNP in prostate androgen-regulated mucin-like protein 1 (PARM1) was not associated with fertility. Overall, results show that variation in response to hCG treatment on fertility depends on parity and interacts with a SNP in COQ9.

Respiratory quotient estimations as additional prognostic tools in early septic shock.

Central venous-to-arterial carbon dioxide difference (PcvaCO2), and its correction by the arterial-to-venous oxygen content difference (PcvaCO2/CavO2) have been proposed as additional tools to evaluate tissue hypoxia. Since the relationship between pressure and content of CO2 (CCO2) might be affected by several factors, some authors advocate for the use of CcvaCO2/CavO2. The aim of the present study was to explore the factors that might intervene in the difference between PcvaCO2/CavO2 and CcvaCO2/CavO2, and to analyze their association with mortality. Observational study in a 30-bed mixed ICU. Fifty-two septic shock patients within the first 24 h of ICU admission were studied. After restoration of mean arterial pressure, hemodynamic and metabolic parameters were evaluated. A total of 110 sets of measurements were performed. Simultaneous PcvaCO2/CavO2 and CcvaCO2/CavO2 values were correlated, but agreement analysis showed a significant proportional bias. The difference between PcvaCO2/CavO2 and CcvaCO2/CavO2 was independently associated with pH, ScvO2, baseline CcvaCO2/CavO2 and hemoglobin. A stepwise regression analysis showed that pH was the single best predictor for the magnitude of such difference, with very limited effect of other variables. At inclusion, variables associated with ICU-mortality were lactate, pH, PcvaCO2/CavO2, and the difference between PcvaCO2/CavO2 and CcvaCO2/CavO2. Initial ScvO2, PcvaCO2, CcvaCO2/CavO2, and cardiac index were not different in survivors and non-survivors. In a population of early septic shock patients, simultaneous values of PcvaCO2/CavO2 and CcvaCO2/CavO2 were not equivalent, and the main determinant of the magnitude of the difference between these two parameters was pH. The PcvaCO2/CavO2 ratio was associated with ICU mortality, whereas CcvaCO2/CavO2 was not.

Is the tiger shark Galeocerdo cuvier a coastal species? Expanding its distribution range in the Atlantic Ocean using at-sea observer data.

The occurrence of tiger shark Galeocerdo cuvier in the Atlantic Ocean was assessed using at-sea observer data from multiple pelagic longline fisheries. Geographic positions of 2764 G. cuvier recorded between 1992 and 2013 and covering a wide area of the Atlantic Ocean were compared with the currently accepted distribution ranges of the species. Most records fell outside those ranges in both the Southern and Northern Hemispheres, which strongly suggests that the distribution range of G. cuvier in the open ocean is considerably larger than previously described.

Modulation of the transient outward current (Ito) in rat cardiac myocytes and human Kv4.3 channels by mefloquine.

The antimalarial drug mefloquine, is known to be a potassium channel blocker, although its mechanism of action has not being elucidated and its effects on the transient outward current (Ito) and the molecular correlate, the Kv4.3 channel has not being studied. Here, we describe the mefloquine-induced inhibition of the rat ventricular Ito and of CHO cells co-transfected with human Kv4.3 and its accessory subunit hKChIP2C by whole-cell voltage-clamp. Mefloquine inhibited rat Ito and hKv4.3+KChIP2C currents in a concentration-dependent manner with a limited voltage dependence and similar potencies (IC50=8.9µM and 10.5µM for cardiac myocytes and Kv4.3 channels, respectively). In addition, mefloquine did not affect the activation of either current but significantly modified the hKv4.3 steady-state inactivation and recovery from inactivation. The effects of this drug was compared with that of 4-aminopyridine (4-AP), a well-known potassium channel blocker and its binding site does not seem to overlap with that of 4-AP.

Network structure of avian mixed-species flocks decays with elevation and latitude across the Andes.

Birds in mixed-species flocks benefit from greater foraging efficiency and reduced predation, but also face costs related to competition and activity matching. Because this cost-benefit trade-off is context-dependent (e.g. abiotic conditions and habitat quality), the structure of flocks is expected to vary along elevational, latitudinal and disturbance gradients. Specifically, we predicted that the connectivity and cohesion of flocking networks would (i) decline towards tropical latitudes and lower elevations, where competition and activity matching costs are higher, and (ii) increase with lower forest cover and greater human disturbance. We analysed the structure of 84 flock networks across the Andes and assessed the effect of elevation, latitude, forest cover and human disturbance on network characteristics. We found that Andean flocks are overall open-membership systems (unstructured), though the extent of network structure varied across gradients. Elevation was the main predictor of structure, with more connected and less modular flocks upslope. As expected, flocks in areas with higher forest cover were less cohesive, with better defined flock subtypes. Flocks also varied across latitude and disturbance gradients as predicted, but effect sizes were small. Our findings indicate that the unstructured nature of Andean flocks might arise as a strategy to cope with harsh environmental conditions. This article is part of the theme issue 'Mixed-species groups and aggregations: shaping ecological and behavioural patterns and processes'.

Complex surface chemistry of 4-mercaptopyridine self-assembled monolayers on Au(111).

The adsorption of 4-mercaptopyridine on Au(111) from aqueous or ethanolic solutions is studied by different surface characterization techniques and density functional theory calculations (DFT) including van der Waals interactions. X-ray photoelectron spectroscopy and electrochemical data indicate that self-assembly from 4-mercaptopyridine-containing aqueous 0.1 M NaOH solutions for short immersion times (few minutes) results in a 4-mercaptopyridine (PyS) self-assembled monolayer (SAM) with surface coverage 0.2. Scanning tunneling microscopy images show an island-covered Au surface. The increase in the immersion time from minutes to hours results in a complete SAM degradation yielding adsorbed sulfur and a heavily pitted Au surface. Adsorbed sulfur is also the main product when the self-assembly process is made in ethanolic solutions irrespective of the immersion time. We demonstrate for the first time that a surface reaction is involved in PyS SAM decomposition in ethanol, a surface process not favored in water. DFT calculations suggest that the surface reaction takes place via disulfide formation driven by the higher stability of the S-Au(111) system. Other reactions that contribute to sulfidization are also detected and discussed.

Increased number of Purkinje cell dendritic swellings in essential tremor.

BACKGROUND AND PURPOSE: Essential Tremor (ET) is among the most prevalent neurologic disorders. Growing clinical and neuro-imaging evidence implicates cerebellar dysfunction in the pathogenesis of ET and emerging postmortem studies have identified structural changes in the cerebellum, particularly in Purkinje cells. In this study we systematically quantified focal Purkinje cell dendritic swellings (DS) in 20 ET vs. 19 control brains. METHODS: In each brain, a standard parasagittal neocerebellar tissue block was harvested. DS were quantified in one 7-µm thick section stained with Luxol Fast Blue/Hematoxylin and Eosin (LH&E) and one section stained with Bielschowsky method. RESULTS: The number of DS were higher in cases than controls by LH&E (1.50 ± 1.79 vs. 0.05 ± 0.23, P = 0.002) and Bielschowsky methods (2.70 ± 3.10 vs. 0.37 ± 0.50, P = 0.002). The number of DS was correlated with the number of torpedoes and marginally inversely correlated with the number of Purkinje cells. CONCLUSION: The current study documents and quantifies an additional structural abnormality in the ET cerebellum, adding to the growing list of such changes in this disease. The mechanisms that underlie this and other structural changes observed in ET are currently unknown, and they deserve additional exploration.

The psychology of LUTS: ICI-RS 2011.

INTRODUCTION: There is undoubtedly a link between lower urinary tract and psychological symptoms. The association is likely to be multifactorial, but whether psychological symptoms are causal or coincidental remains unknown. METHODS: This paper is a result of a think tank session conducted at the ICI-RS meeting 2011 in Bristol and a PubMed search on the subject of psychology and LUTS. RESULTS: Recent population studies have shown that LUTS are prevalent and that psychological morbidity is related to severity of LUTS. Evidence suggests that the psychological status of patients plays a significant role in their symptoms perception. Animal studies have shown the importance of programming and emotional stress on subsequent psychological problems. The link between early events in childhood and resulting LUTS in later life remains uncertain. The possibility that for some patients urinary symptoms could be part of a somatizing process requires further consideration. The possibility that patients' symptoms could be part of a somatization process needs further consideration. Neuroendocrine and neuroimaging studies suggest pathways that may link psychological and lower urinary tract functioning from early life, opening new avenues to better understand the treatment of both symptom groups. CONCLUSION: Whilst psychological morbidity will result from any distressing chronic medical condition the possibility of causation, and or maintenance of LUTS through psychological causes requires further research. It is important to recognize that amongst our younger and older patients suffering with LUTS a significant psychological component exists. Failure to address this may lead to incomplete assessment and consequently inadequate treatment.

In vitro and in vivo assessment of the glycemic index of bakery products: influence of the reformulation of ingredients.

PURPOSE: To evaluate whether the modification of ingredients of two bakery products, muffins and bread, reduces their glycemic index, by means of in vitro and in vivo procedures. METHODS: In vitro and in vivo glycemic index were evaluated for two types of bread and two types of muffins including one standard product for each category. For the in vitro determination, kinetics of starch digestion method was used. For the in vivo procedure, postprandial glucose measured as IAUC was obtained in a group of eighteen healthy volunteers (ten did the test with muffins and eight with breads). RESULTS: In in vitro, a reduction in the expected glycemic index regarding the control muffin was achieved with the partial substitution of wheat flour by a mixture of resistant starch, dextrin and lentil flour. In breads, with the partial substitution of wheat flour by a mixture of resistant starch and dextrins, a decrease in the expected glycemic index was also observed. In in vivo, a reduction in GI was also achieved both in muffin and in bread. All the obtained GI was higher in in vitro method. CONCLUSIONS: Despite the fact that in vitro overestimate in vivo method, the trend in the reduction in GI seems to be similar in both methods. With the substitution assayed, a reduction in the expected glycemic index and the glycemic index were obtained both in muffins and in breads.

Reproductive variables of Urotrygon rogersi (Batoidea: Urotrygonidae): a species with a triannual reproductive cycle in the eastern tropical Pacific Ocean.

Reproductive aspects of the round ray Urotrygon rogersi were studied based on 2005 specimens obtained in the artisanal shrimp fishery operating on the Colombian Pacific coast. Females reached greater maximum total length (L(T) ), disc width (W(D) ) and mass (M) (38·0 cm, 19·9 cm and 348 g) than males (32·5 cm, 17·0 cm and 165 g). Sex ratio of juveniles and adults was 1:1. Clasper length increased rapidly between 10·0 and 12·5 cm W(D) . The smallest mature male measured 10·5 cm W(D) and the largest immature individual 13·7 cm W(D) . Male first maturity was reached at 61·8% of maximum W(D) , and estimated W(D50) was between 11·5 and 11·8 cm. The smallest mature female measured 10·5 cm W(D) ; the size at first maturity was 52·8% of maximum W(D) , and estimated W(D50) was between 11·8 and 12·3 cm. Embryos were found in females ≥ 10·5 cm W(D) and maximum fecundity was three embryos per female (mode = 1) and varied with maternal size. Embryos were found in all months, but three birthing peaks per year were identified and a gestation period of 4-5 months estimated. Based on ovulation time, embryonic growth and parturition dates, a triannual reproductive cycle was inferred for this species, with overlapping ovarian and uterine cycles. These results suggest that U. rogersi has a reproductive strategy based on low fecundity, a rapid reproductive cycle (short ovulation and gestation time), three birth peaks per year and large embryos. This strategy probably allows U. rogersi to withstand the fishing pressure they are subject to on the Colombian Pacific coast. The results also suggest that the study area is an important nursery and reproductive area for this species.

A comparison study of two lower urinary tract symptoms screening tools in clinical practice: the B-SAQ and OAB-V8 questionnaires.

Lower urinary tract symptoms (LUTS) have a detrimental effect on quality-of-life (QoL). However, sufferers are often reluctant to seek help. Screening for LUTS will identify patients with bothersome symptoms who may benefit from treatment and allow patients to self-assess their symptoms and the need for medical intervention, potentially saving costly medical time and reducing long-term morbidity. The aim of this study was to compare the value of two validated questionnaires: the Bladder Control Self Assessment Questionnaire (B-SAQ) and the Overactive Bladder Awareness Tool (OAB-V8) as screening questionnaires in clinical practice. A total of 223 women were recruited prospectively from three centres. Participants completed both questionnaires in the waiting area prior to assessment by a clinician, who completed a symptom evaluation sheet. Data were analysed using receiver operating characteristic curves. Both the B-SAQ and the OAB-V8 performed well in detecting symptoms of OAB and mixed urinary symptoms. The B-SAQ performed better in detecting symptoms of stress incontinence than the OAB-V8. The opportunity to screen for haematuria should never be missed and this is an important omission from the OAB-V8.

[Clinical usefulness of three quantitative D-dimers tests in outpatients with suspected deep vein thrombosis]. / Análisis de 3 métodos para determinar el dímero D en pacientes con sospecha de trombosis venosa profunda.

BACKGROUND AND OBJECTIVE: The diagnostic approach in outpatients with suspected deep vein thrombosis (DVT) of the lower limbs includes D-dimer measurement (DD). Elevated DD is not a diagnostic value for DVT. However, a normal value contributes to ruling out DVT. We do not know the best method to determine DD. Therefore, we have analyzed the clinical utility of three quantitative assays to determine DD in outpatients with suspected DVT. PATIENTS AND METHODS: Consecutive outpatients with suspected DVT of the lower limbs who were referred to the DVT medical consultation were enrolled in the study. We used a diagnostic algorithm that included determining the pretest clinical probability (PCP) (Wells scale), DD level using three different quantitative methods (ELISA mini-VIDAS(®), Acure-care DDMR and DD-Plus). The DVT diagnosis was confirmed by seriated compression ultrasonography of the lower limbs. We analyzed the concordance between the three analytic methods to quantify DD and the characteristics. RESULTS: A total of 306 patients (mean age 60 years, 62% women) with suspected DVT of the lower limbs were included. The compression ultrasonography confirmed the diagnosis of DVT in 23.8% of the patients. Anticoagulation treatment was not performed in patients in whom DVT was ruled out, and no thromboembolic event occurred during the 3 months of follow-up. The best concordance test results were between ELISA mini-VIDAS(®) and Acure-care DDMR assays. Both assays demonstrated elevated sensibility and a negative predictive value. ELISA mini-VIDAS(®) was the best analytic method for the subgroup of patients with low clinical probability. CONCLUSIONS: The ELISA mini-VIDAS(®) method to determine DD rules out DVT in patients with low clinical probability.

Chaotic solitons in the quadratic-cubic nonlinear Schrödinger equation under nonlinearity management.

We analyze the response of rational and regular (hyperbolic-secant) soliton solutions of an extended nonlinear Schrödinger equation (NLSE) which includes an additional self-defocusing quadratic term, to periodic modulations of the coefficient in front of this term. Using the variational approximation (VA) with rational and hyperbolic trial functions, we transform this NLSE into Hamiltonian dynamical systems which give rise to chaotic solutions. The presence of chaos in the variational solutions is corroborated by calculating their power spectra and the correlation dimension of the Poincaré maps. This chaotic behavior (predicted by the VA) is not observed in the direct numerical solutions of the NLSE when rational initial conditions are used. The solitary-wave solutions generated by these initial conditions gradually decay under the action of the nonlinearity management. On the contrary, the solutions of the NLSE with exponentially localized initial conditions are robust solitary-waves with oscillations consistent with a chaotic or a complex quasiperiodic behavior.