How much of the remarkable variation in neuron number within a species is generated by genetic differences, and how much is generated by environmental factors? We address this problem for a single ...population of neurons in the mouse CNS. Retinal ganglion cells of inbred and outbred strains, wild species and subspecies, and F1 hybrids were studied using an unbiased electron microscopic method with known technical reliability. Ganglion cell numbers among diverse types of mice are highly variable, ranging from 32,000 to 87,000. The distribution of all cases (n = 252) is close to normal, with a mean of 58,500 and an SD of 7800. Genetic factors are most important in controlling this variation; 76% of the variance is heritable and up to 90% is attributable to genetic factors in a broad sense. Strain averages have an unanticipated bimodal distribution, with distinct peaks at 55,500 and 63,500 cells. Three pairs of closely related strains have ganglion cell populations that differ by > 20% (10,000 cells). These findings indicate that different alleles at one or two genes have major effects on normal variation in ganglion cell number. Nongenetic factors are still appreciable and account for a coefficient of variation that averages approximately 3.6% within inbred strains and isogenic F1 hybrids. Age- and sex-related differences in neuron number are negligible. Variation within isogenic strains appears to be generated mainly by developmental noise.
Translating promising preclinical pain relief data for novel molecules from drug discovery to positive clinical trial outcomes is challenging. The angiotensin II type 2 (AT
) receptor is a ...clinically-validated target based upon positive proof-of-concept clinical trial data in patients with post-herpetic neuralgia. This trial was conducted because AT
receptor antagonists evoked pain relief in rodent models of neuropathic pain. EMA401 was selected as the drug candidate based upon its suitable preclinical toxicity and safety profile and good pharmacokinetics. Herein, we provide an overview of the discovery, preclinical and clinical development of EMA401, for the alleviation of peripheral neuropathic pain.
The ability of organisms to adapt to sudden extreme environmental changes produces some of the most drastic examples of rapid phenotypic evolution. The Mexican Tetra, Astyanax mexicanus, is abundant ...in the surface waters of northeastern Mexico, but repeated colonizations of cave environments have resulted in the independent evolution of troglomorphic phenotypes in several populations. Here, we present three chromosome-scale assemblies of this species, for one surface and two cave populations, enabling the first whole-genome comparisons between independently evolved cave populations to evaluate the genetic basis for the evolution of adaptation to the cave environment. Our assemblies represent the highest quality of sequence completeness with predicted protein-coding and non-coding gene metrics far surpassing prior resources and, to our knowledge, all long-read assembled teleost genomes, including zebrafish. Whole genome synteny alignments show highly conserved gene order among cave forms in contrast to a higher number of chromosomal rearrangements when compared to other phylogenetically close or distant teleost species. By phylogenetically assessing gene orthology across distant branches of amniotes, we discover gene orthogroups unique to A. mexicanus. When compared to a representative surface fish genome, we find a rich amount of structural sequence diversity, defined here as the number and size of insertions and deletions as well as expanding and contracting repeats across cave forms. These new more complete genomic resources ensure higher trait resolution for comparative, functional, developmental, and genetic studies of drastic trait differences within a species.
Motivated by advances in laser technology and molecular spectroscopy, researchers are again looking for ways to control the temporal evolution of a complex quantum system.
To discover novel therapies that lower IOP by increasing aqueous humor outflow facility, ex vivo ocular perfusion systems provide a valuable tool. However, currently available designs are limited by ...their throughput. Here we report the development of a compact, scalable perfusion system with improved throughput and its validation using bovine and porcine eyes.
At a fixed IOP of 6 mm Hg, flow rate was measured by flow sensors. We validated the system by measuring the outflow responses to Y-39983 (a Rho kinase inhibitor), endothelin-1 (ET-1), ambrisentan (an antagonist for endothelin receptor A ETA), sphigosine-1-phosphate (S1P), JTE-013 (antagonist for S1P receptor 2 S1P2), S-nitroso-N-acetylpenicillamine (SNAP, a nitric oxide NO donor), and 3-Morpholino-sydnonimine (SIN-1, another NO donor).
The instrument design enabled simultaneous measurements of 20 eyes with a footprint of 1 m2. Relative to vehicle control, Y-39983 increased outflow by up to 31% in calf eyes. On the contrary, ET-1 decreased outflow by up to 79%, a response that could be blocked by pretreatment with ambrisentan, indicating a role for ETA receptors. Interestingly, the effect of ET-1 was also inhibited by up to 70% to 80% by pretreatment with NO donors, SNAP and SIN-1. In addition to testing in calf eyes, similar effects of ET-1 and ambrisentan were observed in adult bovine and porcine eyes.
The compact eye perfusion platform provides an opportunity to efficiently identify compounds that influence outflow facility and may lead to the discovery of new glaucoma therapies.
Projections of building electricity consumption are used in multiple fields and for a variety of purposes, from energy utility investment decisions to global climate assessments. Existing approaches ...to modeling building electricity consumption span a range of structural methodologies, spatial resolutions, and temporal scales, potentially leading to divergent projections. This paper compares how two models with different structures and resolutions respond to a common set of population and climate drivers in the western U.S. The BEND model simulates hourly residential and commercial building electricity consumption at the county scale by weighting the results of simulations of thousands of representative buildings. In contrast, the projected electricity demand in GCAM-USA is determined for each state, year, sector (residential/commercial), and service (e.g., heating, cooling, and others), based on population, income, technology, energy prices, and average annual climate. This paper aggregates the two models’ results to a common resolution: annual residential and commercial building electricity consumption by state. Both models show similar responses to future projected population change and climate change, with population change having the larger impact between the two. Differences are primarily due to how the models capture changes in the aggregate energy efficiency of the building stock as it evolves over time.
•Models of building electricity consumption are methodologically diverse.•Two models of building electricity using very different approaches are compared.•The models mostly agree in their projections of building electricity consumption.•How the efficiency of buildings evolves drives disagreement between the models.
Dairy ultrafiltration permeate (UFP) has been filtered using polyamide nanofiltration (NF) membranes in order to separate potentially valuable calcium and lactose from unwanted monovalent salts. An ...investigation into both the fouling and separation behaviour has been carried out to gather useful information for the optimal selection of operating conditions that maximise efficiency and yield. Supersaturated calcium phosphate in the feed resulted in severe flux decline during filtration at high temperatures and pH due to salt precipitation. Investigation into the ion separation behaviour using sub-saturated model solutions containing KCl and CaCl
2 showed that the addition of the trivalent anions citrate and phosphate changed the pH dependent separation of K
+ and Ca
2+. For the multicomponent system, maximum calcium rejection and minimum potassium rejection occurred at high pH, whereas the opposite was true for calcium in the simpler systems. A possible explanation for this separation behaviour has been proposed which incorporates the membrane charge, represented by the zeta potential, and the speciation of calcium with these anions.
Abstract
Study question
Does in vitro exposure of granulosa cells (GC) to an equivalent hyperinsulinemic (HI) dose of insulin alter key signalling pathways, potentially slowing follicle growth?
...Summary answer
Exposure of GC to HI dose of insulin down-regulated FSH-mediated phosphorylation of AKT and ERK proteins, which was reversed by metformin but not myoinositol treatment
What is known already
Antral follicles utilise glycolytic methods of energy production releasing the metabolites pyruvate and lactate, which are then secreted into the follicular fluid or passed directly to the oocyte via gap junctions. Insulin increases GC glucose uptake and cross-talks with FSH-mediated intracellular pathways that determine follicle growth and glucose utilisation. We and others have shown that there is significant perturbation in glucose metabolism (as measured by reduced lactate production) in cultured luteinised GC (GLC) from women with anovulatory PCOS, compared to those with normal ovaries or polycystic ovarian morphology (PCOM), that represents both attenuated glucose uptake and reduced glycolytic activity.
Study design, size, duration
KGN-GC were exposed to 1000ng/ml insulin for 48h (mimicking prolonged HI in vivo) with/without metformin 10-7M or myo-inositol 25mM; followed by acute stimulation with FSH (10ng/ml at 15 and 30 mins) (n = 4-5). GLCs obtained from women with normal ovaries (n = 5), PCOM (n = 7) and PCOS (n = 2) were serum-starved and exposed acutely to insulin (10ng/ml for 30mins), glucose starved (20mins) and treated with a radiolabelled 2-deoxy-glucose mix. Fasting serum glucose and insulin were measured in PCOS women.
Participants/materials, setting, methods
Western blotting on proteins from KGN-GC was performed using antibodies against total and phosphorylated forms of AKT and ERK. Densitometry measurements of phospho:total forms were taken from individual treatments, normalised to loading controls (either β-actin/Vinculin) and then untreated controls. GLCs were lysed and uptake of 14C-2DG (non-metabolised) measured and expressed as percentage uptake of 14C-2DG in the insulin-treated normalised to untreated/control cells (100%). Insulin resistance (IR) was calculated from fasting glucose and insulin using HOMA2.
Main results and the role of chance
Chronic exposure of KGNs to high dose insulin significantly down-regulated acute FSH-mediated phosphorylation of AKT and ERK (ANOVA *p=0.03, multiple post-hoc t-tests p < 0.05, two-tailed), without altering insulin receptor levels. Addition of metformin to FSH treatment, significantly reversed the HI-induced reduction in pAKT levels and enhanced it further (*p=0.02) but had no effect on pERK levels. Myoinositol had no effect on either pAKT or pERK levels. These findings demonstrated that HI directly inhibited FSH signalling events downstream of the insulin receptor in GC, but that metformin could counteract some of these detrimental effects. The 2-deoxy form of glucose is taken up into the cell but not metabolised further. Measurement of 14C-2DG showed that acute stimulation with post-prandial levels of insulin of GLCs taken from normal and PCOM women significantly increased glucose uptake (*p<0.05, ratio of paired t-test, 2-tailed); whereas GLCs from women with PCOS and insulin resistance were unable to respond to insulin and take up glucose. Therefore, GLCs from insulin resistant women with PCOS retain their phenotype in vitro and this is not an artefact, since GLCs from normal and PCOM also subjected to the same stimulation regime were able to take up glucose in response to insulin stimulation.
Limitations, reasons for caution
A clear limitation is the small number of insulin resistant women with PCOS. In addition, human GLCs were collected after hormonal stimulation.
Wider implications of the findings
Women with IR, HI and PCOS undergoing ART may require extra gonadotrophin stimulation to grow and mature sufficient eggs. Additionally, the oocyte quality maybe compromised, as indicated by reduced glucose uptake. Treatment of women with metformin but not myoinositol may improve the dysregulated FSH-signalling pathways brought about by HI.
Trial registration number
not applicable
The products formed from the degradation of the blister agent sulfur mustard bis(2-chloroethyl) sulfide on concrete were identified using gas chromatography with mass spectrometry detection (GC/MSD),
...1H NMR, 2D
1H–
13C NMR and
13C solid state magic angle spinning (SSMAS) NMR. In situ and extraction experiments were performed. Sulfur mustard was detected in the in situ
13C SSMAS samples for 12 weeks, whereas less than 5% of the sulfur mustard was detected in extracts from the concrete monoliths after 8 days. Sulfonium ions and (2-chloroethylthio)ethyl ether (T) were observed on the in situ samples after a period of 12 weeks, whereas vinyl species and bis(2-chloroethyl) sulfoxide were observed in the extracts of the concrete monoliths within 24
h. The differences between the extraction and the SSMAS data indicated that the sulfur mustard existed in the concrete in a non-extractable form prior to its degradation. Extraction methods alone were not sufficient to identify the products; methods to identify the presence of non-extractable degradation products were also required.
Neuropathic pain is a large unmet medical need. The angiotensin II type 2 (AT2) receptor is a target with promising data in rodent models of peripheral neuropathic pain. The AT2 receptor has ...attracted attention on the basis of human data from a proof‐of‐concept clinical trial showing that oral EMA401, a highly selective, peripherally restricted, small molecule AT2 receptor antagonist, at 100 mg twice‐daily for 4 weeks, alleviated postherpetic neuralgia, an often intractable type of peripheral neuropathic pain.
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