Dysregulation of granulosal bone morphogenetic protein receptor 1B density is associated with reduced ovarian reserve and the age-related decline in human fertility

Reproductive ageing is linked to the depletion of ovarian primordial follicles, which causes an irreversible change to ovarian cellular function and the capacity to reproduce. The current study aimed to profile the expression of bone morphogenetic protein receptor, (BMPR1B) in 53 IVF patients exhibiting different degrees of primordial follicle depletion. The granulosa cell receptor density was measured in 403 follicles via flow cytometry. A decline in BMPR1B density occurred at the time of dominant follicle selection and during the terminal stage of folliculogenesis in the 23-30 y good ovarian reserve patients. The 40+ y poor ovarian reserve patients experienced a reversal of this pattern. The results demonstrate an association between age-induced depletion of the ovarian reserve and BMPR1B receptor density at the two critical time points of dominant follicle selection and pre-ovulatory follicle maturation. Dysregulation of BMP receptor signalling may inhibit the normal steroidogenic differentiation required for maturation in older patients.

A low mortality, high morbidity reduced intensity status epilepticus (RISE) model of epilepsy and epileptogenesis in the rat

Animal models of acquired epilepsies aim to provide researchers with tools for use in understanding the processes underlying the acquisition, development and establishment of the disorder. Typically, following a systemic or local insult, vulnerable brain regions undergo a process leading to the development, over time, of spontaneous recurrent seizures. Many such models make use of a period of intense seizure activity or status epilepticus, and this may be associated with high mortality and/or global damage to large areas of the brain. These undesirable elements have driven improvements in the design of chronic epilepsy models, for example the lithium-pilocarpine epileptogenesis model. Here, we present an optimised model of chronic epilepsy that reduces mortality to 1% whilst retaining features of high epileptogenicity and development of spontaneous seizures. Using local field potential recordings from hippocampus in vitro as a probe, we show that the model does not result in significant loss of neuronal network function in area CA3 and, instead, subtle alterations in network dynamics appear during a process of epileptogenesis, which eventually leads to a chronic seizure state. The model’s features of very low mortality and high morbidity in the absence of global neuronal damage offer the chance to explore the processes underlying epileptogenesis in detail, in a population of animals not defined by their resistance to seizures, whilst acknowledging and being driven by the 3Rs (Replacement, Refinement and Reduction of animal use in scientific procedures) principles.

Transcendental Brauer groups of products of CM elliptic curves

Let L be a number field and let E/L be an elliptic curve with complex multiplication by the ring of integers O_K of an imaginary quadratic field K. We use class field theory and results of Skorobogatov and Zarhin to compute the transcendental part of the Brauer group of the abelian surface ExE. The results for the odd order torsion also apply to the Brauer group of the K3 surface Kum(ExE). We describe explicitly the elliptic curves E/Q with complex multiplication by O_K such that the Brauer group of ExE contains a transcendental element of odd order. We show that such an element gives rise to a Brauer-Manin obstruction to weak approximation on Kum(ExE), while there is no obstruction coming from the algebraic part of the Brauer group.

Bad reduction of genus three curves with complex multiplication

Let C be a smooth, absolutely irreducible genus 3 curve over a number field M. Suppose that the Jacobian of C has complex multiplication by a sextic CM-field K. Suppose further that K contains no imaginary quadratic subfield. We give a bound on the primes p of M such that the stable reduction of C at p contains three irreducible components of genus 1.

Shadow lines in the arithmetic of elliptic curves

Let E/Q be an elliptic curve and p a rational prime of good ordinary reduction. For every imaginary quadratic field K/Q satisfying the Heegner hypothesis for E we have a corresponding line in E(K)\otimes Q_p, known as a shadow line. When E/Q has analytic rank 2 and E/K has analytic rank 3, shadow lines are expected to lie in E(Q)\otimes Qp. If, in addition, p splits in K/Q, then shadow lines can be determined using the anticyclotomic p-adic height pairing. We develop an algorithm to compute anticyclotomic p-adic heights which we then use to provide an algorithm to compute shadow lines. We conclude by illustrating these algorithms in a collection of examples.

OX133, a monoclonal antibody recognizing protein-bound N-ethylmaleimide for the identification of reduced disulfide bonds in proteins

In vivo, enzymatic reduction of some protein disulfide bonds, allosteric disulfide bonds, provides an important level of structural and functional regulation. The free cysteine residues generated can be labeled by maleimide reagents, including biotin derivatives, allowing the reduced protein to be detected or purified. During the screening of monoclonal antibodies for those specific for the reduced forms of proteins, we isolated OX133, a unique antibody that recognizes polypeptide resident, N-ethylmaleimide (NEM)-modified cysteine residues in a sequence-independent manner. OX133 offers an alternative to biotin-maleimide reagents for labeling reduced/alkylated antigens and capturing reduced/alkylated proteins with the advantage that NEM-modified proteins are more easily detected in mass spectrometry, and may be more easily recovered than is the case following capture with biotin based reagents.

Structural features of condensed tannins affect in vitro ruminal methane production and fermentation characteristics

An in vitro study was conducted to investigate the effects of condensed tannins (CT) structural properties, i.e. average polymer size (or mean degree of polymerization); percentage of cis flavan-3-ols and percentage of prodelphinidins in CT extracts on methane production (CH4) and fermentation characteristics. CT were extracted from eight plants in order to obtain different CT types: black currant leaves, goat willow leaves, goat willow twigs, pine bark, red currant leaves, sainfoin plants, weeping willow catkins and white clover flowers. They were analysed for CT content and CT composition by thiolytic degradation, followed by HPLC analysis. Grass silage was used as a control substrate. Condensed tannins were added to the substrate at a concentration of 40 g/kg, with or without polyethylene glycol (+ or −PEG 6000 treatment) to inactivate tannins, and then incubated for 72 h in mixed buffered rumen fluid from three different lactating dairy cows per run. Total cumulative gas production (GP) was measured by an automated gas production system. During the incubation, 12 gas samples (10 μl) were collected from each bottle headspace at 0, 2, 4, 6, 8, 12, 24, 30, 36, 48, 56 and 72 h of incubation and analyzed for CH4. A modified Michaelis–Menten model was fitted to the CH4 concentration patterns and model estimates were used to calculate total cumulative CH4 production (GPCH4). Total cumulative gas production and GPCH4 curves were fitted using biphasic and monophasic modified Michaelis-Menten models, respectively. Addition of PEG increased GP, GPCH4, and CH4 concentration compared to the −PEG treatment. All CT types reduced GPCH4 and CH4 concentration. All CT increased the half time of GP and GPCH4. Moreover, all CT decreased the maximum rate of fermentation for GPCH4 and rate of substrate degradation. The correlation between CT structure and GPCH4 and fermentation characteristics showed that the proportion of prodelphinidins within CT had the largest effect on fermentation characteristics, followed by average 27 polymer size and percentage of cis-flavan-3-ols.

Emergence dynamics of barnyardgrass and jimsonweed from two depths when switching from conventional to reduced and no-till conditions

A cylinder experiment was conducted in northern Greece during 2005 and 2006 to assess emergence dynamics of barnyardgrass (Echinochloa crus-galli (L.) Beauv.) and jimsonweed (Datura stramonium L.) in the case of a switch from conventional to conservation tillage systems (CT). Emergence was surveyed from two burial depths (5 and 10 cm) and with simulation of reduced tillage (i.e. by soil disturbance) and no-till conditions. Barnyardgrass emergence was significantly affected by burial depth, having greater emergence from 5 cm depth (96%) although even 78% of seedlings emerged from 10 cm depth after the two years of study. Emergence of barnyardgrass was stable across years from the different depths and tillage regimes. Jimsonweed seeds showed lower germination than barnyardgrass during the study period, whereas its emergence was significantly affected by soil disturbance having 41% compared to 28% without disturbance. A burial depth x soil disturbance interaction was also determined, which showed higher emergence from 10 cm depth with soil disturbance. Jimsonweed was found to have significantly higher emergence from 10 cm depth with soil disturbance in Year 2. Seasonal emergence timing of barnyardgrass did not vary between the different burial depth and soil disturbance regimes, as it started in April and lasted until end of May in both years. Jimsonweed showed a bimodal pattern, with first emergence starting end of April until mid-May and the second ranging from mid-June to mid-August from 10 cm burial depth and from mid-July to mid-August from 5 cm depth, irrespective of soil disturbance in both cases.

Effects of rain shelter or simulated rain during grain filling and maturation on subsequent wheat grain quality in the UK

The effects of simulated additional rain (ear wetting, 25 mm) or of rain shelter imposed at different periods after anthesis on grain quality at maturity and the dynamics of grain filling and desiccation were investigated in UK field-grown crops of wheat (Triticum aestivum L., cvar Tybalt) in 2011 and in 2012 when June–August rainfall was 255.0 and 214.6 mm, respectively, and above the decadal mean (157.4 mm). Grain filling and desiccation were quantified well by broken-stick regressions and Gompertz curves, respectively. Rain shelter for 56 (2011) or 70 d (2012) after anthesis, and to a lesser extent during late maturation only, resulted in more rapid desiccation and hence progress to harvest maturity whereas ear wetting had negligible effects, even when applied four times. Grain-filling duration was also affected as above in 2011, but with no significant effect in 2012. In both years, there were strong positive associations between final grain dry weight and duration of filling. The treatments affected all grain quality traits in 2011: nitrogen (N) and sulphur (S) concentrations, N:S ratio, sodium dodecyl sulphate (SDS) sedimentation volume, Hagberg Falling Number (HFN), and the incidence of blackpoint. Only N concentration and blackpoint were affected significantly by treatments in 2012. Rain shelter throughout grain filling reduced N concentration, whereas rain shelter reduced the incidence of blackpoint and ear wetting increased it. In 2011, rain shelter throughout reduced S concentration, increased N:S ratio and reduced SDS. Treatment effects on HFN were not consistent within or between years. Nevertheless, a comparison between the extreme treatment means in 2012 indicated damage from late rain combined with ear wetting resulted in a reduction of c. 0.7 s in HFN/mm August rainfall, whilst that between samples taken immediately after ear wetting at harvest maturity or 7 d later suggested recovery from damage to HFN upon re-drying in planta. Hence, the incidence of blackpoint was the only grain quality trait affected consistently by the diverse treatments. The remaining aspects of grain quality were comparatively resilient to rain incident upon developing and maturing ears of cvar Tybalt. No consistent temporal patterns of sensitivity to shelter or ear wetting were detected for any aspect of grain quality.

Effects of Cobalt Chloride in Junctional Epithelium and Reduced Enamel Epithelium of the Rat Maxillary First Molar

Cobalt is one of the main components of cast metal alloys broadly used in dentistry. It is the constituent of 45 to 70% of numerous prosthetic works. There are evidences that metal elements cause systemic and local toxicity. The purpose of the present study was to evaluate the effects of cobalt on the junctional epithelium and reduced enamel epithelium of the first superior molar in rats, during lactation. To do this, 1-day old rats were used, whose mothers received 300mg of cobalt chloride per liter of distilled water in the drinker, during lactation. After 21 days, the rat pups were killed with an anesthetic overdose. The heads were separated, fixed in ""alfac"", decalcified and embedded in paraffin. Frontal sections stained with hematoxylin and eosin were employed. Karyometric methods allowed to estimate the following parameters: biggest, smallest and mean diameters, D/d ratio, perimeter, area, volume, volume/area ratio, eccentricity, form coefficient and contour index. Stereologic methods allow to evaluate: cytoplasm/nucleus ratio, cell and cytoplasm volume, cell number density, external surface/basal membrane ratio, thickness of the epithelial layers and surface density. All the collected data were subjected to statistic analysis by the non-parametric Wilcoxon-Mann-Whitney test. The nuclei of the studied tissues showed smaller values after karyometry for: diameters; perimeter, area, volume and volume/area ratio. Stereologically, it was observed, in the junctional epithelium and in the reduced enamel epithelium, smaller cells with scarce cytoplasm, reflected in the greater number of cells per mm3 of tissue. In this study, cobalt caused epithelial atrophy, indicating a direct action on the junctional and enamel epithelium.

Physical and sensory properties of regular and reduced-fat pound cakes with added amaranth flour

The sum of wheat flour and corn starch was replaced by 10, 20, or 30% whole amaranth flour in both conventional (C) and reduced fat (RF) pound cakes. and the effects on physical and sensory properties of the cakes were investigated. RF presented 33% fat reduction. The increasing amaranth levels darkened crust and crumb of cakes, which decreased color acceptability. Fresh amaranth-containing cakes had similar texture characteristics to (he controls, evaluated both instrumentally and sensorially. Sensory evaluation revealed that replacement by 30% amaranth flour decreased C cakes overall acceptability scores, clue to its lower specific volume and darker color. Amaranth flour levels had no significant effect on overall acceptability of RF cakes. Hence, the sum of wheat flour and corn starch could be successfully replaced by up to 20% amaranth flour in C and up to 30% in RF pound cakes without negatively affecting sensory quality in fresh cakes. Moisture losses for all the cakes were similar, approximate to 1% per day during storage. After six days of storage, both C and RF amaranth-containing cakes had higher hardness and chewiness values than control cakes. Further experiments involving sensory evaluation during storage are necessary to determine the exact limit of amaranth flour replacement.

Loss of Methylation at H19 DMD Is Associated with Biallelic Expression and Reduced Development in Cattle Derived by Somatic Cell Nuclear Transfer

Although cloning of mammals has been achieved successfully, the percentage of live offspring is very low because of reduced fetal size and fewer implantation sites. Recent studies have attributed such pathological conditions to abnormal reprogramming of the donor cell used for cloning. The inability of the oocyte to fully restore the differentiated status of a somatic cell to its pluripotent and undifferentiated state is normally evidenced by aberrant DNA methylation patterns established throughout the genome during development to blastocyst. These aberrant methylation patterns are associated with abnormal expression of imprinted genes, which among other genes are essential for normal embryo development and gestation. We hypothesized that embryo loss and low implantation rates in cattle derived by somatic cell nuclear transfer (SCNT) are caused by abnormal epigenetic reprogramming of imprinted genes. To verify our hypothesis, we analyzed the parental expression and the differentially methylated domain (DMD) methylation status of the H19 gene. Using a parental-specific analysis, we confirmed for the first time that H19 biallelic expression is tightly associated with a severe demethylation of the paternal H19 DMD in SCNT embryos, suggesting that these epigenetic anomalies to the H19 locus could be directly responsible for the reduced size and low implantation rates of cloned embryos in cattle.

CYP3A5*3A allele is associated with reduced lowering-lipid response to atorvastatin in individuals with hypercholesterolemia

Background: The cytochrome P450 isoenzyme 3A5 (CYP3A5) has an important role on biotransformation of xenobiotics. CYP3A5 SNPs have been associated with variations on enzyme activity that can modify the metabolism of several drugs. Methods: In order to evaluate the influence of CYP3A5 variants on response to lowering-cholesterol drugs, 139 individuals with hypercholesterolemia were selected. After a wash-out period of 4 weeks, individuals were treated with atorvastatin (10 mg/day/4 weeks). Genomic DNA was extracted by a salting-out procedure. CYP3A5*3C, CYP3A5*6 and CYP3A5*1D were analyzed by PCR-RFLP and DNA sequencing. Results: >Frequencies of the CYP3A5*3C and CYP3A5*1D alleles were lower in individuals of African descent (*3C: 47.8% and *1D: 55.2%) than in non-Africans (*3C: 84.9% and *1D 84.8%, p<0.01). Non-Africans carrying *3A allele (*3C and *1D combined alleles) had lower total and LDL-cholesterol response to atorvastatin than non-*3A allele carriers (p<0.05). Conclusion: CYP3A5*3A allele is associated with reduced cholesterol-lowering response to atorvastatin in non-African individuals. (C) 2008 Elsevier B.V. All rights reserved.

The SARS algorithm: detrending CoRoT light curves with Sysrem using simultaneous external parameters

Surveys for exoplanetary transits are usually limited not by photon noise but rather by the amount of red noise in their data. In particular, although the CoRoT space-based survey data are being carefully scrutinized, significant new sources of systematic noises are still being discovered. Recently, a magnitude-dependant systematic effect was discovered in the CoRoT data by Mazeh et al. and a phenomenological correction was proposed. Here we tie the observed effect to a particular type of effect, and in the process generalize the popular Sysrem algorithm to include external parameters in a simultaneous solution with the unknown effects. We show that a post-processing scheme based on this algorithm performs well and indeed allows for the detection of new transit-like signals that were not previously detected.