Automated mass spectrometric analysis of urinary and plasma serotonin

Serotonin emerges as crucial neurotransmitter and hormone in a growing number of different physiologic processes. Besides extensive serotonin production previously noted in patients with metastatic carcinoid tumors, serotonin now is implicated in liver cell regeneration and bone formation. The aim was to develop a rapid, sensitive, and highly selective automated on-line solid-phase extraction method coupled to high-performance liquid chromatography–tandem mass spectrometry (XLC-MS/MS) to quantify low serotonin concentrations in matrices such as platelet-poor plasma and urine. Fifty microliters plasma or 2.5 μL urine equivalent were pre-purified by automated on-line solid-phase extraction, using weak cation exchange. Chromatography of serotonin and its deuterated internal standard was performed with hydrophilic interaction chromatography. Mass spectrometric detection was operated in multiple reaction monitoring mode using a quadrupole tandem mass spectrometer with positive electrospray ionization. Serotonin concentrations were determined in platelet-poor plasma of metastatic carcinoid patients (n = 23) and healthy controls (n = 22). Urinary reference intervals were set by analyzing 24-h urine collections of 120 healthy subjects. Total run-time was 6 min. Intra- and inter-assay analytical variation were <10%. Linearity in the 0–7300 μmol/L calibration range was excellent (R2 > 0.99). Quantification limits were 30 and 0.9 nmol/L in urine and plasma, respectively. Platelet-poor serotonin concentrations in metastatic carcinoid patients were significantly higher than in controls. The urinary reference interval was 10–78 μmol/mol creatinine. Serotonin analysis with sensitive and specific XLC-MS/MS overcomes limitations of conventional HPLC. This enables accurate quantification of serotonin for both routine diagnostic procedures and research in serotonin-related disorders

Drain blocking has limited short-term effects on greenhouse gas fluxes in a Molinia caerulea dominated shallow peatland (article)

This is the final version. Available on open access from Elsevier via the DOI in this recordThe dataset associated with this article is available in ORE at https://doi.org/10.24378/exe.2723Drained peatlands dominated by purple moor grass (Molinia caerulea) are widespread in the UK and Western Europe. Although substantial carbon stores may be present in these peatlands, in this degraded state they are not currently acting as carbon sinks. Therefore, M.caerulea dominated peatlands have been identified as potential sites for ecohydrological restoration to tackle the current climate emergency. However, at present little is known about whether ditch blocking can raise water tables and promote the recovery of bog plant species, and the subsequent effects on carbon sequestration in these peatlands. To investigate the potential for restoration, we measured changes in water table depth, vegetation composition, photosynthesis at 1000 μmol Photons m−2 s−1 (PG1000), ecosystem respiration (REco) and partitioned below-ground respiration in two M.caerulea dominated peatlands in which drainage ditches had been blocked located in Exmoor National Park, southwest England. Measurements were made in two headwater catchments at ⅛, ¼ and ½ of the distance between adjacent drainage ditches at four control-restored paired sites, during the growing seasons pre- (2012) and post- (2014, 2016 & 2018) restoration. Restoration had a small but significant (p = 0.009) effect on water table depths however, this did not result in a significant change in vegetation composition (p > 0.350). Ecosystem respiration increased in both the control and restored locations following restoration however, this increase was significantly smaller (p = 0.010) at the restored locations, possibly due to a similarly reduced increase in photosynthesis, although this change was not significant (p = 0.116). Below-ground respiration showed no significant changes following restoration. This research illustrates how degraded these shallow peatlands are, and raises concerns that ditch blocking alone may not bring about the high and stable water tables required to perturb the existing Molinia caerulea-dominated ecosystem and substantially alter the carbon balance. Additional restoration measures may be required.South West Water (SWW)University of ExeterTechnology Strategy Board CouncilNatural Environment Research Council (NERC

Expression of trk in MAH Cells Lacking the p75 Low-Affinty Nerve Growth Factor Receptor is Sufficient to Permit Nerve Growth Factor-Induced Differentiation to Postmitotic Neurons

We have transfected MAH cells, an immortalized sympathoadrenal progenitor cell line, with a plasmid encoding the 140-kDa Trk protein, a nerve growth factor (NGF) receptor with protein-tyrosine kinase activity. NGF promotes neurite outgrowth and proliferation from such cells, indicating that Trk is sufficient to mediate such responses in the absence of significant levels of the endogenous 75-kDa low-affinity NGF receptor (p75). These initial NGF responses are indistinguishable from those evoked by basic fibroblast growth factor (bFGF). However, NGF is sufficient to promote terminal differentiation of a {approx}8% of trk-transfected MAH cells to postmitotic, NGF-dependent neurons, whereas all cells eventually die in medium with bFGF. Other environmental signals (such as depolarization or ciliary neurotrophic factor) can cooperate with NGF to enhance production of postmitotic NGF-dependent neurons in trk-transfected MAH cells. The terminal differentiation of sympathetic neurons thus involves sequential and cooperative actions of different growth and neurotrophic factors, as well as cell-intrinsic changes in the response to these factors

Observations of Mercury's northern cusp region with MESSENGER's Magnetometer

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/95510/1/grl29143.pd

Developmental Exposure to Polychlorinated Biphenyls Influences Stroke Outcome in Adult Rats

BackgroundThe "developmental origins of adult disease" hypothesis was originally derived from evidence linking low birth weight to cardiovascular diseases including stroke. Subsequently, it has been expanded to include developmental exposures to environmental contaminants as risk factors for adult onset disease.ObjectiveOur goal in this study was to test the hypothesis that developmental exposure to poly-chlorinated biphenyls (PCBs) alters stroke outcome in adults.MethodsWe exposed rats to the PCB mixture Aroclor 1254 (A1254) at 0.1 or 1 mg/kg/day in the maternal diet throughout gestation and lactation. Focal cerebral ischemia was induced at 6-8 weeks of age via middle cerebral artery occlusion, and infarct size was measured in the cerebral cortex and striatum at 22 hr of reperfusion. PCB congeners were quantified in brain tissue by gas chromatography with microelectron capture detection, and cortical and striatal expression of Bcl2 and Cyp2C11 were quantified by quantitative reverse transcriptase-polymerase chain reaction.ResultsDevelopmental exposure to A1254 significantly decreased striatal infarct in females and males at 0.1 and 1 mg/kg/day, respectively. Predominantly ortho-substituted PCB congeners were detected above background levels in brains of adult females and males exposed to A1254 at 1 but not 0.1 mg/kg/day. Effects of developmental A1254 exposure on Bcl2 and Cyp2C11 expression did not correlate with effects on infarct volume.ConclusionOur data provide proof of principle that developmental exposures to environmental contaminants influence the response of the adult brain to ischemic injury and thus represent potentially important determinants of stroke susceptibility

Ion kinetic properties in Mercury's pre-midnight plasma sheet

With data from the Fast Imaging Plasma Spectrometer sensor on the MErcury Surface, Space ENvironment, GEochemistry, and Ranging spacecraft, we demonstrate that the average distributions for both solar wind and planetary ions in Mercury’s pre-midnight plasma sheet are well-described by hot Maxwell-Boltzmann distributions. Temperatures and densities of the H+ ranges ~1–10 cm3 and ~5–30 MK, respectively, maintain thermal pressures of ~1 nPa. The dominant planetary ion, Na+ abundances with respect to H+ and exhibit mass-proportional ion temperatures, indicative of a reconnection-dominated heating in the magnetosphere. Conversely, planetary ion species are accelerated to similar average energies greater by a factor of ~1.5 than that of H+ acceleration in an electric potential, consistent with the presence of a strong centrifugal acceleration process in Mercury’s magnetosphere

Unwinding of a cholesteric liquid crystal and bidirectional surface anchoring

We examine the influence of bidirectional anchoring on the unwinding of a planar cholesteric liquid crystal induced by the application of a magnetic field. We consider a liquid crystal layer confined between two plates with the helical axis perpendicular to the substrates. We fixed the director twist on one boundary and allow for bidirectional anchoring on the other by introducing a high-order surface potential. By minimizing the total free energy for the system, we investigate the untwisting of the cholesteric helix as the liquid crystal attempts to align with the magnetic field. The transitions between metastable states occur as a series of pitchjumps as the helix expels quarter or half-turn twists, depending on the relative sizes of the strength of the surface potential and the bidirectional anchoring. We show that secondary easy axis directions can play a significant role in the unwinding of the cholesteric in its transition towards a nematic, especially when the surface anchoring strength is large

Thymus transplantation for complete DiGeorge syndrome: European experience

Background: Thymus transplantation is a promising strategy for the treatment of athymic complete DiGeorge syndrome (cDGS). Methods: Twelve patients with cDGS were transplanted with allogeneic cultured thymus. Objective: To confirm and extend the results previously obtained in a single centre. Results: Two patients died of pre-existing viral infections without developing thymopoeisis and one late death occurred from autoimmune thrombocytopaenia. One infant suffered septic shock shortly after transplant resulting in graft loss and the need for a second transplant. Evidence of thymopoeisis developed from 5-6 months after transplantation in ten patients. The median (range) of circulating naïve CD4 counts (x10663 /L) were 44(11-440) and 200(5-310) at twelve and twenty-four months post-transplant and T-cell receptor excision circles were 2238 (320-8807) and 4184 (1582 -24596) per106 65 T-cells. Counts did not usually reach normal levels for age but patients were able to clear pre-existing and later acquired infections. At a median of 49 months (22-80), eight have ceased prophylactic antimicrobials and five immunoglobulin replacement. Histological confirmation of thymopoeisis was seen in seven of eleven patients undergoing biopsy of transplanted tissue including five showing full maturation through to the terminal stage of Hassall body formation. Autoimmune regulator (AIRE) expression was also demonstrated. Autoimmune complications were seen in 7/12 patients. In two, early transient autoimmune haemolysis settled after treatment and did not recur. The other five suffered ongoing autoimmune problems including: thyroiditis (3); haemolysis (1), thrombocytopaenia (4) and neutropenia (1). Conclusions: This study confirms the previous reports that thymus transplantation can reconstitute T cells in cDGS but with frequent autoimmune complications in survivors

A Review of Multi- Compartment Infectious Disease Models

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/156488/2/insr12402.pdfhttp://deepblue.lib.umich.edu/bitstream/2027.42/156488/1/insr12402_am.pd

Abnormalities in autonomic function in obese boys at-risk for insulin resistance and obstructive sleep apnea.

Study objectivesCurrent evidence in adults suggests that, independent of obesity, obstructive sleep apnea (OSA) can lead to autonomic dysfunction and impaired glucose metabolism, but these relationships are less clear in children. The purpose of this study was to investigate the associations among OSA, glucose metabolism, and daytime autonomic function in obese pediatric subjects.MethodsTwenty-three obese boys participated in: overnight polysomnography; a frequently sampled intravenous glucose tolerance test; and recordings of spontaneous cardiorespiratory data in both the supine (baseline) and standing (sympathetic stimulus) postures.ResultsBaseline systolic blood pressure and reactivity of low-frequency heart rate variability to postural stress correlated with insulin resistance, increased fasting glucose, and reduced beta-cell function, but not OSA severity. Baroreflex sensitivity reactivity was reduced with sleep fragmentation, but only for subjects with low insulin sensitivity and/or low first-phase insulin response to glucose.ConclusionsThese findings suggest that vascular sympathetic activity impairment is more strongly affected by metabolic dysfunction than by OSA severity, while blunted vagal autonomic function associated with sleep fragmentation in OSA is enhanced when metabolic dysfunction is also present