Palle Bratholm

Lymphocyte norepinephrine and epinephrine, but not plasma catecholamines predict lymphocyte cAMP production.

Endogenous norepinephrine (NE) and epinephrine (E) were demonstrated in lymphocytes isolated from peripheral venous blood. In 13 young subjects lymphocyte NE and E averaged 14.3 and 1.9 pg per 10(7) cells, respectively. The ratio NE/E was similar in plasma and in lymphocytes. Highly significant correlations were obtained between lymphocyte NE and E on the one hand and cAMP in lymphocytes on the other both in the basal state and after stimulation with isoproterenol. In a group of elderly subjects lymphocyte NE concentration was significantly reduced in long-term smokers as compared to non-smokers (7 and 35 pg/10(7) cells, respectively), whereas plasma NE was increased in smokers. Addition of exogenous NE or propranolol to blood samples did not change lymphocyte NE concentration in in vitro experiments. Variability in endogenous lymphocyte concentration of E in 9 young subjects, correlated with concomitant changes in number of NK(CD3-CD56+) cells and cAMP. It is concluded that endogenous lymphocyte NE and E concentrations in healthy subjects reflected basal cAMP production in lymphocytes and lymphocyte subset composition.

Effect of Oxytocin Receptor Blockade on Rat Myometrial Responsiveness to Prostaglandin F2α

Abstract In the present study we have shown that the genetic expression of prostaglandin (PG)F2α receptor (R) and cyclooxygenase (COX)-2 increases in laboring rat myometrium. This finding was associated with a relatively weak contractile in vitro response (Emax) of isolated uterine strips when challenged with PGF2α. Five days postpartum PGF2α-R mRNA values exceeded those during labor while COX-2 mRNA was reduced to preparturient values. Maximal contractility of isolated strips stimulated with PGF2α at this time was enhanced and EC50 decreased. Oxytocin treatment of estrogen-primed nonpregnant rats down-regulated uterine contractile responsiveness to PGF2α, leaving mRNA values for this receptor unchanged, whereas oxytocin receptor blockade with atosiban (an oxytocin receptor antagonist) left Emax unaltered. In contrast, atosiban treatment of pregnant rats resulted in a 2.5-fold increase in Emax and a considerably reduced EC50 during labor when compared to untreated delivering rats. The increased contractile ability was associated with a threefold increase in PGF2α-R mRNA production, indicating that the regulation by atosiban of the PGF2α-induced response is exerted at the genetic level. Based on the present data we suggest that 1) PGF2α-R stimulation may not primarily exert a contracting role in the normally delivering myometrium, and 2) the presence of the PGF2α-R system in rat myometrium may explain the apparent functional redundancy of the oxytocinergic system during the process of birth in animals lacking oxytocin or where the oxytocin receptor is blocked. In this context PGF2α receptor stimulation may, in the absence of oxytocin receptor stimulation, exert the contractile forces needed for proper propulsion of the fetus.

Simultaneous determination of glucose, triglycerides, urea, cholesterol, albumin and total protein in human plasma by Fourier transform infrared spectroscopy: direct clinical biochemistry without reagents.

OBJECTIVE Direct measurement of chemical constituents in complex biologic matrices without the use of analyte specific reagents could be a step forward toward the simplification of clinical biochemistry. Problems related to reagents such as production errors, improper handling, and lot-to-lot variations would be eliminated as well as errors occurring during assay execution. We describe and validate a reagent free method for direct measurement of six analytes in human plasma based on Fourier-transform infrared spectroscopy (FTIR). DESIGN AND METHODS Blood plasma is analyzed without any sample preparation. FTIR spectrum of the raw plasma is recorded in a sampling cuvette specially designed for measurement of aqueous solutions. For each analyte, a mathematical calibration process is performed by a stepwise selection of wavelengths giving the optimal least-squares correlation between the measured FTIR signal and the analyte concentration measured by conventional clinical reference methods. The developed calibration algorithms are subsequently evaluated for their capability to predict the concentration of the six analytes in blinded patient samples. RESULTS The correlation between the six FTIR methods and corresponding reference methods were 0.87<R(2)<1.00. The interassay imprecision meets international quality criteria for all the six analytes. The linearity of the FTIR methods extends over the clinically significant concentration ranges. Visible hemolysis and icterus have some influence on the measurements. Plasma samples can be stored at 2-8°C for at least 8 days before the analysis. CONCLUSIONS The developed FTIR methods use a simple and robust technology to achieve stable and accurate results that meet international quality criteria for the measurement of glucose, triglycerides, urea, cholesterol, albumin and total protein in human plasma.

Regulation of TSH Receptor Autoantibodies by a long Non-Coding RNA (Heg) and Cdk1- A Review

Aims: A substantialpartofthegenome istranscribedinnon-codingRNAs. We review ourfinding of along non-coding RNA (designatedHeg)in mononuclearcells(MNC) and regulation of TSHreceptorautoantibodies(TRAb). Results:The Heg RNA transcriptinMNC isnegatively correlatedwithTRAb inpatients withearlyand untreatedGravesdisease.In treatedpatients andincontrolsHeg correlatednegativelywithCD14 mRNA. Transfection studieswithfragments ofHeg added toMNC (exogenous Heg)decreased CD14 mRNA in MNC and increasedgene expressionofRIG-I,TLR7 and IFN-�≥ .Heg islikelyto activate TLR7 receptors. CD14 isa co-receptorofTLR7. Decrease ingene expressionofCD14 afterHeg isa signof differentiationofMNC todendriticcells.Thismay reduce surfaceexpressionof CD14, cytokineresponses and the responsiveness to TSH receptorantigens. Thusthe relationshipbetween TRAb and lncHeg RNA ismost likelyexplainedbyreceptorcross- interference.Cdk1 mRNA (an indexofcellcycleactivity)ispositivelyrelatedwithTRAb. Cdk1 mRNA and TRAb butnotHeg decreased significantlyduringantithyroidtreatment. Cdk1 decreased tovaluesbelownormal. Conclusion: Thus bothHeg RNA and Cdk1 may regulatethe level ofTRAb but by two different mechanisms.