Search

Opioid analgesics are commonly used for acute low back pain and neck pain, but supporting efficacy data are scarce. We aimed to investigate the efficacy and safety of a judicious short course of an opioid analgesic for acute low back pain and neck pain.

GLPG1972/S201086 is a disintegrin and metalloproteinase with thrombospondin motif‐5 (ADAMTS‐5) inhibitor in development as an osteoarthritis disease‐modifying therapy. We report the safety, tolerability, pharmacokinetics, and pharmacodynamics (turnover of plasma/serum ARGS‐aggrecan neoepitope fragments [ARGS]) of GLPG1972 in 3 randomized, double‐blind, placebo‐controlled phase 1 trials. Study A, a first‐in‐human trial of single (≤2100 mg [fasted] and 300 mg [fed]) and multiple (≤1050 mg once daily [fed]; 14 days) ascending oral (solution) doses, investigated GLPG1972 in healthy men (N = 41; NCT02612246). Study B investigated multiple ascending oral (tablet) doses of GLPG1972 (≤300 mg once daily [fed]; 4 weeks) in male and female participants with osteoarthritis (N = 30; NCT03311009). Study C investigated single (Japanese: ≤1500 mg; White: 300 mg [fasted]) and multiple (Japanese, ≤1050 mg once daily; White, 300 mg once daily [fed]; 14 days) ascending oral (tablet) doses of GLPG1972 in healthy Japanese and White men (N = 88). The pharmacokinetic profile of GLPG1972 was similar between healthy participants and participants with osteoarthritis, with low to moderate interindividual variability. GLPG1972 was rapidly absorbed (median time to maximum concentration, 4 hours), and eliminated with a mean apparent terminal elimination half‐life of ≈10 hours. Steady state was achieved within 2 days of dosing, with minimal accumulation. Steady‐state plasma exposure after 300 mg of GLPG1972 showed no or minor differences between populations. Area under the plasma concentration–time curve (56.8‐67.6 μg · h/mL) and time to maximum concentration (4 hours) were similar between studies. Urinary excretion of GLPG1972 (24 hours) was low (<11%). Multiple dosing significantly reduced ARGS levels vs baseline at all time points for all doses vs placebo. GLPG1972 was generally well tolerated at all doses.

Noninvasive prenatal screening (NIPS) using cell-free DNA has transformed prenatal care. Belgium was the first country to implement and fully reimburse NIPS as a first-tier screening test offered to all pregnant women. A consortium consisting of all Belgian genetic centers report the outcome of two years genome-wide NIPS implementation.

Idiopathic pulmonary fibrosis (IPF) causes irreversible loss of lung function. People with IPF have increased concentrations of autotaxin in lung tissue and lysophosphatidic acid (LPA) in bronchoalveolar lavage fluid and exhaled condensate. GLPG1690 (Galapagos, Mechelen, Belgium) is a novel, potent, selective autotaxin inhibitor with good oral exposure. We explored the effects of GLPG1690 in patients with IPF.

Objectives: BELGICA-STROKE is a longitudinal study to enhance the use of online cardiovascular risk prediction scores based on the SCORE 10-year risk estimates for fatal cardiovascular disease (adapted for Belgium) and the Framingham 10-year stroke risk and to evaluate their impact on the cardiovascular risk profile of hypertensive patients. Methods and baseline characteristics are described here.Design: Prospective, multicenter study in primary care.Methods: General practitioners (N = 810) recruited consecutive hypertensive patients aged >40 years who were not at blood pressure goal and assessed them every 4 months. The estimated 10-year risks for fatal cardiovascular disease and stroke were available on a secured, specially designed study website. The calculated risk profile of a patient was modifiable by adding treatment goals in order to increase awareness and motivation of both physician and patient. An automated feedback on goal-level attainment and both cardiovascular risk scores was provided.Results: Mean age of the 15,744 patients was 66.3 years: 51.9% were men, 77.8% had excess weight, 19.4% were smokers, and 25.9% had diabetes. Left ventricle hypertrophy was present in 20.0%, atrial fibrillation in 5.8%. Mean blood pressure was 153.8/88.2 mmHg, mean cholesterol 211.5 mg/dl. Most patients (89.2%) received antihypertensive medication, of which 36.9% was monotherapy. Mean estimated 10-year stroke risk was 19.1%, and mean estimated 10-year fatal cardiovascular disease risk 5.9%.Conclusions: The 10-year estimated stroke and fatal cardiovascular disease risks were moderate to high in hypertensive patients not at goal blood pressure, emphasizing the importance of global cardiovascular risk factor assessment.

S100B is a calcium binding protein from astrocytes that regulates protein phosphorylation by binding to substrates and protein kinases. S100B might also regulate protein phosphatases and this was investigated for protein phosphatase 2B (calcineurin). The results indicate that S100B (5–10 μM) increased the activity of both purified and cytoskeletal calcineurin in a Ca2-dependent manner. This effect was blocked by a specific inhibitor of calcineurin activity, but not by TRTK-12 (an inhibitor of S100B binding to other protein targets). The present results and the known co-localization of S100B and calcineurin in the astrocyte cytoskeleton suggest that S100B may play a role in the phosphorylation state of cytoskeletal proteins.

The mechanism of carrier-mediated sucrose uptake by the dermal transfer cells of developing Vicia fabaL. cotyledons was studied using excised cotyledons and isolated transfer cell protoplasts. Addition of sucrose resulted in a transitory alkalinization of the bathing solution whereas additions of glucose, fructose or raffinose had no effect. Dissipating the proton motive force by exposing cotyledons and isolated transfer cell protoplasts to an alkaline pH, carbonylcyanide m-chlorophenylhydrazone, weak acids (propionic acid and 5,5′-dimethyl-oxazolidine-2,4-dione) or tetraphenylphos-phonium ion resulted in a significant reduction of sucrose uptake. The ATPase inhibitors, erythrosin B (EB), diethylstilbestrol (DES) and N,N′-dicyclohexylcarbodiimide (DCCD) were found to abolish the sucrose-induced medium alkanization as well as reduce sucrose uptake. Cytochemical localization of the ATPase, based on lead precipitation, demonstrated that the highest activity was present in the plasma membranes located in wall ingrowth regions of the dermal transfer cells. The presence of a transplasma-membrane redox system was detected by the extracellular reduction of the electron acceptor, hexacyanoferrate III. The reduction of the ferric ion was coupled to a release of protons. The redox-induced proton extrusion was abolished by the ATPase inhibitors EB, DES and DCCD suggesting that proton extrusion was solely through the H+-ATPase. Based on these findings, it is postulated that cotyledonary dermal transfer cells take up sucrose by a proton symport mechanism with the proton motive force being generated by a H +-ATPase. Sucrose uptake by the storage parenchyma and inner epidermal cells of the cotyledons did not exhibit characteristics consistent with sucrose-proton symport.

Two experimental systems were developed to study the uptake of sucrose by the dermal transfer cells of developing cotyledons of Vicia faba L. First, the in-vivo state was approximated by short-term (10 min) incubation of whole cotyledons in [14C]sucrose solutions. Under these conditions, a minimum of 67% of the 14C label entered the dermal transfer cell complex. Of this, at least 40% crossed the plasma membranes of the epidermal transfer cells. Second, a protocol was developed to enzymatically isolate and purify dermal transfer cell protoplasts. The yields of the transfer cell protoplasts were relatively low and their preparation incurred a significant loss of plasma membrane. However, the protoplasts remained viable up to 24 h following purification and proved to be a suitable system to verify transport properties observed with whole cotyledons. Using these two experimental systems, it was established that [14C]sucrose uptake by the dermal transfer cells exhibited features consistent with mediated energy-dependent transport. This included saturation kinetics, competition for uptake between structurally similar molecules, and inhibition of uptake by p-chloromercuribenzenesulfonic acid and several other metabolic inhibitors. For comparative purposes, sugar uptake by the storage parenchyma of the Vicia cotyledons was also examined. In contrast to the dermal transfer cell complex, sucrose uptake by the storage parenchyma displayed characteristics consistent with simple diffusion.

Abstract.: Cytosolic NADP-dependent isocitrate dehydrogenase (cyt-NADP-ICDH; EC 1.1.1.42) has been suggested to play a major role in the production of 2-oxoglutarate, an important precursor for amino acid synthesis. Using an antisense RNA approach under the control of the cauliflower mosaic virus 35S promoter, transgenic potato plants were created in which NADP-ICDH activity was reduced to 8% of the wild-type level in leaves. Residual activity was almost completely due to mitochondrial and chloroplastic NADP-ICDH isoforms. Activity staining after non-denaturing polyacrylamide gel electrophoresis revealed the complete absence of a major activity band in leaves of antisense plants. No differences in growth or development, including flower formation and tuber yield, were observed between transgenic and wild-type plants. Photosynthesis and respiration were also unchanged. Levels of amino acids were the same in wild-type and cyt-NADP-ICDH antisense plants, even when accumulation of amino acids was induced by incubation of detached leaves in tap water in the dark (`induced senescence'). Consistent with a reduction in NADP-ICDH activity, however, were slight increases in the levels of isocitrate (up to 2.5-fold) and citrate (up to 2-fold). 2-Oxoglutarate was not reduced. Our data indicate that potato plants can cope with a severe reduction in cyt-NADP-ICDH activity without major shifts in growth and metabolism.

Seed coats of Phaseolus vulgaris L. unload photosynthetic products, mineral ions and acid into the apoplastic space surrounding the embryo. We report measurements, on detached seed coats, of the rates of unloading of photosynthates, ions and acid at different external pH and in the presence of treatments intended to alter the rate of proton pumping. We also report measurements of membrane potential difference (PD) and of cytoplasmic pH under the same conditions, measurements which have allowed us to validate the treatments we used and to investigate functional relationships between membrane processes. A chemiosmotic model of the seed-coat cell membrane is proposed, in which sucrose efflux and acid efflux are both driven by the proton pump. Sucrose efflux is proposed to occur by sucrose/proton antiport driven by the proton-motive force (PMF), and acid efflux to occur by pumped protons accompanied by a passive efflux of anions. We use our measurements to estimate the net efflux of sucrose on the antiporter and the total efflux of protons on the pump. We have tested the model by using experimental treatments designed to manipulate the pump rate as the independent variable. Under these conditions, and assuming the model is correct, the pump rate determines the cytoplasmic pH. Over the range covered by our experiments the net sucrose efflux is dependent on external and cytoplasmic pH, the latter having the major role. The efflux of acid, under the same treatments, depends primarily on the proton pump rate, and was found to be well fitted by a quadratic function of pump rate. This means that, as pump rate increases, an increasing proportion of the pump output is used by acid efflux and a decreasing proportion by sucrose antiport. The membrane PD, although an important component of the PMF, does not appear to function in rate control of net sucrose or of acid efflux, since neither efflux is correlated with membrane PD under our treatments which vary the pump rate. The PD correlates well with external potassium concentration, and seems largely determined by the diffusion of potassium ions and anions.

The cells responsible for the photosynthate efflux from coats of developing seed of Vicia faba L. and Phaseolus vulgaris L. were elucidated using known properties of the efflux mechanism. Sensitivity of sucrose efflux to NEM and high potassium concentrations was retained by seed-coat halves of Phaseolus following pectinase removal of the branch parenchyma cell layer. In contrast, removal of the thin-walled parenchyma transfer cell layer from Vicia seed-coat halves abolished this sensitivity. The membrane-impermeant thiol-binding fluorochrome, qBBr, selectively stained the surface of the thin-walled parenchyma transfer cells. This phenomenon was inhibited by the slowly permeable sul-phydryl agent, PCMBS, indicating that the plasma membranes of these cells are enriched in sulphydryl groups characteristic of membrance porter proteins. On the basis that carrier-mediated sucrose efflux from seed coats appears to be proton coupled, the putative plasma membrane H+-ATPase was used as a marker for the cells responsible for carrier-mediated photosynthate efflux. When seed-coat halves were exposed briefly at pH 8.5 to the weak acid fluorochrome, SRG, the ground parenchyma and thin-walled parenchyma transfer cell layers selectively accumulated the dye. The apparent low pH environment in the walls of these cells that renders SRG membrane permeant appeared to be maintained by a VAN-sensitive proton pump. The findings with SRG were corroborated by the cyto-chemical localization of plasma membrane ATPase activity to the ground parenchyma and thin-walled parenchyma transfer cells using precipitation of cerium phosphate. Together, our observations provide qualified support for the conclusion that carrier-mediated photosynthate efflux from coats of Phaseolus and Vicia seed is primarily restricted to the ground parenchyma and thin-walled parenchyma transfer cell layers, respectively.

In order to develop a tentative model of the mechanism of photosynthate efflux from the vascular region of Vicia faba L. seed coats, wash-out experiments were performed after removal of the embryo. The sulphydryl group modifiers, pCMBS and NEM, reduced 14C-photosynthate efflux by 40% and 50%, respectively. Their inhibitory effect could be prevented or reduced (in the latter case) by including DTT in the bathing solution. Maltose competed with sucrose for efflux; a concentration of 300 mol m−3 inhibited 14C-photosynthate release by 35%. The cations K+ , Na+ Mg2+ and TPP+ enhanced efflux significantly, whereas the countenon Cl− had no effect. The presence of the protonophore CCCP (0·1 mol m−3) led to a reduction of efflux by 50% net proton extrusion dropped by 34%. To a lesser extent, an efflux inhibition was also achieved by decreasing the cytoplasmic pH with the weak acid DM0. In contrast, alterations in the external pH caused only a feeble response. The ATPase inhibitor, EB, decreased photosynthate efflux and H+ extrusion. DES reduced efflux slightly, presumably by affecting ATPase activity as well as energy metabolism. Based on these findings, it is proposed that a sucrose/proton antiport mechanism could be responsible for photosynthate efflux from Vicia faba seed coats.