Genetic relationships, serological cross-reaction and cross-protection between H1N2 and other influenza A virus subtypes endemic in European pigs

This study examines the genetic relationships between the recently emerged H1N2 swine influenza virus and viruses of H1N1 and H3N2 subtypes, and the extent of protection against H1N2 challenge in pigs immune after infection or vaccination with the other subtypes. There was low amino acid homology (70.4-71.9%) in the haemagglutinin (HA) gene between H1N1 viruses used for primary infection or vaccination and the H1N2 challenge strain, with 94-99 amino acid changes between these viruses involving all five antigenic sites. The NA genes of H3N2 viruses used for primary infection or vaccination showed higher amino acid homology with H1N2 (88.3-92.6%), while nucleoprotein (95.5-96.3% nucleotide identity) and matrix (96.8-98.4%) genes were most conserved between the three subtypes. Pigs immune as a result of intranasal inoculation with either H1N1 or H3N2 showed partial clinical protection against H1N2 challenge, and nasal virus excretion was 2 days shorter than in naive pigs. Moreover, dually infected (H1N1 + H3N2)-immune pigs showed complete clinical protection and H1N2 virus replication in the lungs and nasal secretions was either undetectable or markedly reduced. In contrast, a double vaccination with a commercial H1N1 and H3N2-based vaccine did not protect against H1N2 challenge. Haemagglutination inhibition (HI) or virus neutralisation (VN) tests of swine sera revealed little if any antigenic cross-reactivity between subtypes. These data suggest that serum HI or VN antibodies are not essential in heterosubtypic protection, but that mucosal or cellular immunity are probably involved. It is still unknown whether this type of cross-subtype protection will also occur in infection-immune pigs in the field.     

In vitro studies, pharmacokinetic studies and clinical use of a high purity double virus inactivated FVIII/VWF concentrate (Immunate) in the treatment of von Willebrand disease

Patients with type 2 and 3 von Willebrand disease (VWD) are treated with factor VIII/VWF concentrate in case of bleeding or surgery. Immunate (Baxter, Vienna, Austria) is a double virus inactivated FVIII/VWF concentrate and is registered in several countries for patients with VWD with reduced FVIII levels. We performed an in vitro, a pharmacokinetic and a clinical study to evaluate Immunate in VWD. In vitro studies showed a significant variation in VWF levels in 9 different batches. The median (range) values (in IU/mL) were 1.10 (0.98-1.30) for FVIII:C, 1.34 (0.95-1.61) for VWF:Ag, 0.60 (0.27-1.08) for VWF:CBA and 0.73 (0.59-0.94) for VWF:RCo. The relatively low VWF activity is mainly due to the lack of high molecular weight multimers (HMWM), as determined by electrophoresis. A pharmacokinetic study showed, based on a content of FVIII:C of 1 U/mL, in vivo recoveries (%) of 106 (56-150) (median and range) for FVIII:C, 105 (62-187) for VWF:Ag, 25 (7-41) for VWF:CBA and 43 (11-76) for VWF:RCo. Half-lives were 14.1 h (7.4-36.9) for FVIII:C, 10.8 h (7.7-26.2) for VWF:Ag, 15.3 h (7.8-44.6) for VWF:CBA and 16.4 h (4.2-26.5) for VWF:RCo. In a clinical study efficacy was determined after infusion given before surgery or dental extractions in ten patients. In two patients the hemostatic response was classified as inadequate. In conclusion, there is a wide variability in VWF concentration and activity in various batches of Immunate. In the clinical study in which the dosage was based on FVIII:C contents of the concentrate, two out of ten patients had an insufficient haemostatic response. Therefore dosing of Immunate dosing should not be based on FVIII:C levels, but should be based on VWF activity of the individual batches. Future studies using a VWF activity-guided dosage regimen have to be performed to establish the efficacy of Immunate in the treatment of von Willebrand disease.     

JNJ16259685, a highly potent, selective and systemically active mGlu1 receptor antagonist

We examined the pharmacological profile of (3,4-dihydro-2H-pyrano[2,3]b quinolin-7-yl) (cis-4-methoxycyclohexyl) methanone (JNJ16259685). At recombinant rat and human metabotropic glutamate (mGlu) 1a receptors, JNJ16259685 non-competitively inhibited glutamate-induced Ca2+ mobilization with IC50 values of 3.24+/-1.00 and 1.21+/-0.53 nM, respectively, while showing a much lower potency at the rat and human mGlu5a receptor. JNJ16259685 inhibited [3H]1-(3,4-dihydro-2H-pyrano[2,3-b]quinolin-7-yl)-2-phenyl-1-ethanone ([3H]R214127) binding to membranes prepared from cells expressing rat mGlu1a receptors with a Ki of 0.34+/-0.20 nM. JNJ16259685 showed no agonist, antagonist or positive allosteric activity toward rat mGlu2, -3, -4 or -6 receptors at concentrations up to 10 microM and did not bind to AMPA or NMDA receptors, or to a battery of other neurotransmitter receptors, ion channels and transporters. In primary cerebellar cultures, JNJ16259685 inhibited glutamate-mediated inositol phosphate production with an IC50 of 1.73+/-0.40 nM. Subcutaneously administered JNJ16259685 exhibited high potencies in occupying central mGlu1 receptors in the rat cerebellum and thalamus ( ED50=0.040 and 0.014 mg/kg, respectively). These data show that JNJ16259685 is a selective mGlu1 receptor antagonist with excellent potencies in inhibiting mGlu1 receptor function and binding and in occupying the mGlu1 receptor after systemic administration.     

Prenatal diagnosis of facial clefting as part of the oculo-auriculo-vertebral spectrum

We report a rare facial cleft (type 2 according to the Tessier classification) as the first presenting echographic sign of the oculo-auriculo-vertebral spectrum (OAVS) (Goldenhar syndrome). Associated malformations included a left lateral cleft with macrostomia, left ear hypoplasia, left preauricular tag, single umbilical artery, hyposegmentation of the left lung and imperforatio ani.     

Small molecule crystallography in drug design

Crystal structures of small molecules (i.e. isolated ligands) are a source of valuable structural information helpful in the process of drug design (pharmacophore model elaborations, 3D QSAR, docking, and de novo design). Indeed, structural data obtained from small molecules crystallography can approach ligand-receptor binding by providing unique structural features both about the conformation (internal geometry) of the ligand (s) and about the intermolecular interaction potentially occurring within the active site of a target (enzyme/receptor). Small molecule crystal structure databases can also be used in three dimensional search to identify new drug candidates. Future development in small molecule crystallography (e.g. powder diffraction) should also provide original solutions to complex problems related to polymorphism.     

Ring chromosome 20 epilepsy syndrome in children: electroclinical features

Ring chromosome 20 mosaicism is associated with dysmorphic features, mental retardation, and intractable seizures, including recurrent episodes of nonconvulsive status epilepticus. The authors' findings in four children, all without dysmorphic features, indicate that mental deterioration and frequent subtle nocturnal frontal lobe seizures, associated with a characteristic EEG pattern, represent prominent additional clinical features not previously described in this syndrome. This emphasizes the importance of full-night video-EEG in children with frontal lobe seizures and cognitive deterioration.     

Acute pulmonary hypertension causes depression of left ventricular contractility and relaxation

BACKGROUND AND OBJECTIVE: The haemodynamic effects of acute pulmonary hypertension can be largely attributed to ventricular interdependence during diastole. However, there is evidence that the two ventricles also interact during systole. The aim of the present study was to examine the effects of acute pulmonary hypertension on both components of left ventricular systole, i.e. contraction and relaxation, using load-independent indices. METHODS: Ten pigs were instrumented with biventricular conductance catheters, a pulmonary artery flow probe and a high-fidelity pulmonary pressure catheter. Haemodynamic measurements were performed in baseline conditions and during stable pulmonary vasoconstriction induced by the thromboxane analogue U46619. Contractility was quantified using the end-systolic pressure-volume and preload recruitable stroke work relationships. The tau-end-systolic pressure relationship was used to assess load-dependency of relaxation. RESULTS: Acute pulmonary hypertension caused a decrease in the slope of the left ventricular preload recruitable stroke work relationship (from 6.64 +/- 1.7 to 5.19 +/- 1.9, mean +/- SD; P < 0.05), a rightward shift of the end-systolic pressure-volume relationship (P < 0.05), and an increase in the slope of the tau-end-systolic pressure relationship (from -0.15 +/- 0.5 to 0.35 +/- 0.17; P < 0.05). The diastolic chamber stiffness constant of both ventricles increased during pulmonary hypertension (P < 0.05). CONCLUSIONS: In the present model, acute pulmonary hypertension impairs left ventricular contractile function and relaxing properties. The present study provides additional evidence that, besides the well-known diastolic ventricular cross talk, systolic ventricular interaction may play a significant role in the haemodynamic consequences of acute pulmonary hypertension.     

Body-water compartments measured by bio-electrical impedance spectroscopy in patients with chronic obstructive pulmonary disease

It was previously demonstrated that single frequency bio-electrical impedance (BIA) measurement at 50 kHz is a useful method to assess total body water (TBW) in patients with chronic obstructive pulmonary disease (COPD). In the present study it was examined whether bio-electrical impedance spectroscopy (BIS) could predict extracellular water (ECW) and improve the prediction of TBW in these patients. TBW and ECW (corrected bromide space) were measured by deuterium and bromide dilution. In 37 COPD patients prediction equations were obtained using BIS (5-500 kHz) measurements, and these were cross validated in a second group of 40 COPD patients. All patients were in a clinically stable condition. TBW predicted by BIS was not significantly different from actual TBW and demonstrated a comparable standard error of estimate (SEE) as found previously in healthy subjects (male symbol correlation coefficient: r = 0.88, SEE: 2.3 L, female symbol r = 0.85, SEE: 2.9 L). Predicted ECW using BIS-measurements was not significantly different from measured ECW (male symbol r = 0.75, SEE: 1.4 L, female symbol r = 0.73, SEE: 1.2 L), but the error in the prediction was relatively large and the correlation between predicted and actual ECW relatively low compared to most studies in healthy subjects. Predicted TBW using BIS was comparable to actual TBW, but presented no improvement of the prediction of TBW using BIA at 50 kHz and a patient specific regression equation. The error of the prediction of ECW by BIS limits the ability to predict fluid shifts in individual patients with clinically stable COPD.