Polymorphisms of PAI-1 and platelet GP Ia may associate with impairment of renal function and thrombocytopenia in Puumala hantavirus infection

Introduction

Puumala virus (PUUV) infection is a viral hemorrhagic fever with renal syndrome (HFRS) characterized by thrombocytopenia and acute impairment of renal function. We aimed to assess whether genetic polymorphisms of platelet antigens together with those of von Willebrand factor (VWF) and plasminogen activator inhibitor (PAI-1) correlate with disease severity.Patients and methods172 consecutive hospital-treated patients with serologically confirmed acute PUUV infection were included. Platelet glycoprotein (GP) IIIa T > C (rs5918), GP Ia T > C (rs1126643), GP Ib C > T (rs6065), GP VI T > C (rs1613662), VWF A > G (rs1063856) and PAI-1 A > G (rs2227631) were genotyped. The associations of the rarer alleles with variables reflecting the severity of the disease were analyzed.

Results

PAI-1 G-carriers had higher maximum creatinine level compared with the non-carriers (median 213 μmol/l, range 60-1499 μmol/l vs. median 122 μmol/l, range 51-1156 μmol/l, p = 0.01). The GG-genotypes had higher creatinine levels than GA- and AA-genotypes (medians 249 μmol/l, 204 μmol/l and 122 μmol/l, respectively, p = 0.03). Polymorphisms of GP VI and VWF associated with lower creatinine levels during PUUV infection. The minor C-allele of GP Ia associated with lower platelet counts (median 44 × 10⁹/l, range 20-90 × 10⁹/l vs median 64 × 10⁹/l, range 3-238 × 10⁹/l; p = 0.02).

Conclusions

Polymorphism of PAI-1, a major regulator of fibrinolysis, has an adverse impact on the outcome of kidney function in PUUV-HFRS. Platelet collagen receptor GP Ia polymorphism associates with lower platelet count.     

Stimulus evaluation, event preparation, and motor action planning in young patients with mild spastic cerebral palsy: an event-related brain potential study

The study investigated stimulus evaluation time, event preparation, and motor action planning of patients with mild spastic cerebral palsy and a peer control group in the age range of 9 to 18 years. To this end, participants were carrying out a stimulus recognition task. Findings indicated an overall slowness and inaccurate reaction time performance of the patient group. An event-related potential analysis revealed that the stimulus evaluation processing, indexed by the parietal P300, was intact in the group of patients. Also event preparation and action planning, indexed by respectively the frontal late contingent negative variation and the frontal P2, were intact in the group of patients. It was concluded that patients' motor slowness reflected poor motor execution processes.     

Photocrosslinked poly(ester anhydride)s for peptide delivery: Effect of oligomer hydrophobicity on PYY3-36 delivery

The treatment for many diseases can be improved by developing more efficient peptide delivery technologies, for example, biodegradable polymers. In this work, photocrosslinked poly(ester anhydride)s based on functionalized poly(ε-caprolactone) oligomers were investigated for their abilities to achieve controlled peptide delivery. The effect of oligomer hydrophobicity on erosion and peptide release from poly(ester anhydride)s was evaluated by developing a sustained subcutaneous delivery system for an antiobesity drug candidate, peptide YY3-36 (PYY3-36). Oligomer hydrophobicity was modified with alkenylsuccinic anhydrides containing a 12-carbon alkenyl chain. PYY3-36 was mixed as a solid powder with methacrylated poly(ester anhydride) precursors, and this mixture was photocrosslinked at room temperature to form an implant for subcutaneous administration in rats. The oligomer hydrophobicity controlled the polymer erosion and PYY3-36 release as the increased hydrophobicity via the alkenyl chain prolonged polymer erosion in vitro and sustained in vivo release of PYY3-36. In addition, photocrosslinked poly(ester anhydride)s increased the bioavailability of PYY3-36 by up to 20-fold in comparison with subcutaneous administration of solution, evidence of remarkably improved delivery. In conclusion, this work demonstrates the suitability of photocrosslinked poly(ester anhydride)s for use in peptide delivery.     

Removal of large muscle artifacts from transcranial magnetic stimulation-evoked EEG by independent component analysis

We present two techniques utilizing independent component analysis (ICA) to remove large muscle artifacts from transcranial magnetic stimulation (TMS)-evoked EEG signals. The first one is a novel semi-automatic technique, called enhanced deflation method (EDM). EDM is a modification of the deflation mode of the FastICA algorithm; with an enhanced independent component search, EDM is an effective tool for removing the large, spiky muscle artifacts. The second technique, called manual method (MaM) makes use of the symmetric mode of FastICA and the artifactual components are visually selected by the user. In order to evaluate the success of the artifact removal methods, four different quality parameters, based on curve comparison and frequency analysis, were studied. The dorsal premotor cortex (dPMC) and Broca’s area (BA) were stimulated with TMS. Both methods removed the very large muscle artifacts recorded after stimulation of these brain areas. However, EDM was more stable, less subjective, and thus also faster to use than MaM. Until now, examining lateral areas of the cortex with TMS—EEG has been restricted because of strong muscle artifacts. The methods described here can remove those muscle artifacts, allowing one to study lateral areas of the human brain, e.g., BA, with TMS—EEG.     

Exploring the Solid-Form Landscape of Pharmaceutical Hydrates: Transformation Pathways of the Sodium Naproxen Anhydrate-Hydrate System

Purpose

To understand the transformation pathways amongst anhydrate/hydrate solid forms of sodium naproxen and to highlight the importance of a polymorphic dihydrate within this context.

Methods

Multi-temperature dynamic vapour sorption (DVS) analysis combined with variable-humidity X-ray powder diffraction (XRPD) to establish the transformation pathways as a function of temperature and humidity. XRPD and thermogravimetric analysis (TGA) to characterise bulk samples. Monitoring of in-situ dehydration using solid-state ¹³C CP/MAS spectroscopy.

Results

At 25°C, anhydrous sodium naproxen (AH) transforms directly to one dihydrate polymorph (DH-II). At 50°C, AH transforms stepwise to a monohydrate (MH) then to the other dihydrate polymorph (DH-I). DH-II transforms to a tetrahydrate (TH) more readily than DH-I transforms to TH. Both dihydrate polymorphs transform to the same MH.

Conclusions

The properties of the polymorphic dihydrate control the transformation pathways of sodium naproxen.     

Interactions of sesquiterpenes zederone and germacrone with the human cytochrome P450 system

Misclassification of Curcuma species (family Zingiberaceae) may lead to unwanted human exposure to Curcuma elata sesquiterpenes zederone and germacrone which have caused hepatotoxicity and changes in CYP expression in laboratory animals. We investigated how these compounds interact with the human cytochrome P450 (CYP) system, in order to evaluate their potential for human liver toxicity and herb–drug interactions. We found that both sesquiterpenes (1–30 μM) greatly induced expression of CYP2B6 and CYP3A4 but not CYP1A2 mRNAs in human primary hepatocytes (HPHs). This induction profile correlated with activation of constitutive androstane and pregnane X receptors. Cytotoxicity was also observed in exposed HPHs. CYP inhibition studies with pooled human liver microsomes (HLMs) indicated that zederone and germacrone moderately inhibited CYP2B6 and CYP3A4 activities in vitro, with IC₅₀ values below 10 μM. When zederone was incubated with HLMs and NADPH, one di-epoxide metabolite was formed and by using glutathione trapping, five epoxide-derived conjugates were detected. Germacrone produced two oxidized metabolites and four glutathione conjugates. The results suggest that enzymes in HLMs convert sesquiterpenes into reactive, electrophilic compounds which may be causative for the reported liver injuries. These findings provide insight on the safety and drug–herb interactions of the Curcuma species.     

Pharmacological Characterisation of a Structurally Novel α2C ‐Adrenoceptor Antagonist ORM‐10921 and its Effects in Neuropsychiatric Models

The α₂‐adrenoceptors (ARs) are important modulators of a wide array of physiological responses. As only a few selective compounds for the three α₂‐AR subtypes (α_2A, α_2B and α_2C) have been available, the pharmacological profile of a new α_2C‐selective AR antagonist ORM‐10921 is reported. Standard in vitro receptor assays and antagonism of α₂, and α₁‐AR agonist ‐evoked responses in vivo were used to demonstrate the α_2C‐AR selectivity for ORM‐10921 which was tested in established behavioural models related to schizophrenia and cognitive dysfunction with an emphasis on pharmacologically induced hypoglutamatergic state by phencyclidine or MK‐801. The Kb values of in vitro α_2C‐ AR antagonism for ORM‐10921 varied between 0.078–1.2 nM depending on the applied method. The selectivity ratios compared to α_2A‐AR subtype and other relevant receptors were 10‐100 times in vitro. The in vivo experiments supported its potent α_2C‐antagonism combined with only a weak α_2A‐antagonism. In the pharmacodynamic microdialysis study, ORM‐10921 was found to increase extracellular dopamine levels in prefrontal cortex in the baseline conditions. In the behavioural tests, ORM‐10921 displayed potent antidepressant and antipsychotic‐like effects in the forced swimming test and prepulse‐inhibition models analogously with the previously reported results with structurally different α_2C‐selective AR antagonist JP‐1302. Our new results also indicate that ORM‐10921 alleviated the NMDA‐antagonist‐induced impairments in social behaviour and watermaze navigation. This study extends and further validates the concept that α_2C‐AR is a potential therapeutic target in CNS disorders such as schizophrenia or Alzheimer's disease and suggests the potential of α_2C‐antagonism to treat such disorders .