Further DNA sequence microheterogeneity of the HLA-DR4/Dw13 haplotype group: Importance of amino acid position 86 of the DRβ1 chain for T-cell recognition

Using the polymerase chain reaction we have isolated and sequenced cDNA clones corresponding to the polymorphic first domain of the DRβ1 chain from the DR4, “Dw13” cell line, JHa. We have found that the JHa DRβ1 allele differs from previously reported Dw13 alleles by a single amino acid substitution at position 86. The functional relevance of this polymorphism is supported by the reactivity pattern of a T-cell clone, E38. E38 is an alloreactive T-cell clone which reacts with all Dw14 stimulator cells and all Dw13-positive cells tested except the “Dw13”- positive homozygous typing cell line JHa. Inhibition studies with monoclonal antibodies revealed the stimulating determinant to be on DR and not on DQ or DP molecules. These data indicate that position 86 of the DRβ1 chain can play an important role in the formation of determinants recognized by T cells.     

Noncardiogenic pulmonary edema caused by decompression sickness: rapid resolution following hyperbaric therapy

Noncardiogenic pulmonary edema is a recognized but uncommon manifestation of type 2 decompression sickness. It typically occurs within 6 hours of a dive. Because the adult respiratory distress syndrome in this setting is believed to be due to microbubbles in the pulmonary vasculature, recompression in a hyperbaric chamber has been recommended as a form of therapy. A patient developed noncardiogenic pulmonary edema following a seawater dive to 75 feet. There was complete radiologic and clinical resolution within 5 hours of hyperbaric therapy.     

Intertrigo-like eruption caused by pegylated liposomal doxorubicin (PLD)

Pegylated liposomal doxorubicin (PLD) is a drug whose use is increasingly common. It has been associated with a lower rate of haematologic and cardiac side effects than its nonencapsulated form. However, mucocutaneous toxicity is quite frequent and can be severe. Here we provide a case report of a patient who developed an intertrigolike eruption during treatment with PLD.     

Endogenous N-acyl-dopamines induce COX-2 expression in brain endothelial cells by stabilizing mRNA through a p38 dependent pathway

Cerebral microvascular endothelial cells play an active role in maintaining cerebral blood flow, microvascular tone and blood brain barrier (BBB) functions. Endogenous N-acyl-dopamines like N-arachidonoyl-dopamine (NADA) and N-oleoyl-dopamine (OLDA) have been recently identified as a new class of brain neurotransmitters sharing endocannabinoid and endovanilloid biological activities. Endocannabinoids are released in response to pathogenic insults and may play an important role in neuroprotection. In this study we demonstrate that NADA differentially regulates the release of PGE₂ and PGD₂ in the microvascular brain endothelial cell line, b.end5. We found that NADA activates a redox-sensitive p38 MAPK pathway that stabilizes COX-2 mRNA resulting in the accumulation of the COX-2 protein, which depends on the dopamine moiety of the molecule and that is independent of CB₁ and TRPV1 activation. In addition, NADA inhibits the expression of mPGES-1 and the release of PGE₂ and upregulates the expression of L-PGD synthase enhancing PGD₂ release. Hence, NADA and other molecules of the same family might be included in the group of lipid mediators that could prevent the BBB injury under inflammatory conditions and our findings provide new mechanistic insights into the anti-inflammatory activities of NADA in the central nervous system and its potential to design novel therapeutic strategies to manage neuroinflammatory diseases.     

Cord blood banking in London: the first 1000 collections.

The London Cord Blood Bank was established with the aim of collecting, processing and storing 10000 unrelated stem cell donations for the significant number of children in the UK requiring transplantation, for whom a matched unrelated bone marrow donor cannot be found. Collection is performed at two hospitals by dedicated cord blood bank staff after delivery of the placenta. Mothers are interviewed regarding medical, ethnic and behavioural history by nurse counsellors and sign a detailed consent form. Donations are returned to the bank for processing. Volume reduction is undertaken by a simple, closed, semi-automated blood processing system, with excellent recovery of progenitor cells. Units are cryopreserved and stored in the vapour phase of liquid nitrogen. Blood samples from mothers and cord blood donations are tested for the UK mandatory red cell and microbiology markers for blood donors. Donations are typed for HLA-A, B and DR at medium resolution (antigen split) level using sequence-specific oligonucleotide probing and sequence-specific priming techniques. The selection of collection hospitals on the basis of ethnic mix has proven effective, with 41.5% of donations derived from non-European caucasoid donors. Bacterial contamination of collections has been dramatically reduced by implementation of improved umbilical cord decontamination protocols.     

Differences in cholesterol incorporation into mitochondria from hepatoma AS-30D and human term placenta.

Cholesterol transport for steroidogenesis in the human placental mitochondria is an enigma as, contrary to other steroidogenic tissues, the human placenta does not express steroidogenic acute regulatory protein (StAR), a protein known to be required for efficient utilization of cholesterol by adrenal and gonadal mitochondria. These observations suggest the possibility that cholesterol transport in human placental mitochondria involves a similar system to that present in other non-steroidogenic tissues. We studied cholesterol incorporation into mitochondria isolated from AS-30D hepatoma cells and the human placenta. Mitochondria from both sources incorporated cholesterol in vitro. There were no differences in cholesterol incorporation into hepatoma mitochondria treated with or without trypsin. In contrast, the human placental mitochondria treated with trypsin did not incorporate exogenous cholesterol. The presence of ATP increased the uptake of cholesterol by human placental mitochondria. This increase was inhibited by vanadate. These results suggest that cholesterol incorporation into human placental mitochondria is mediated by protein(s).     

Long-lasting modification of reflexes after neonatal nerve injury in the rat

The reflex activity of motoneurones to the extensor digitorum longus (EDL) muscle following sciatic nerve crush during the first 5 days after birth (neonatal crush) or in the adult (adult crush) was studied 3-6 months later, when the axons had reinnervated their target muscles. Electromyograms (EMG) and muscle tension were recorded from the EDL muscle (a physiological flexor) on the injured and uninjured sides. Reflex responses were evoked by stimulation of the common peroneal (CP), the tibial (T) and the sural (S) nerves, ipsilateral and contralateral to the side of injury. In animals which had sustained a neonatal crush, stimulation of branches of the injured sciatic nerve elicited ipsilateral reflex responses that were about 3 times larger than those recorded from the uninjured side or in normal animals. Stimulation of the CP nerve on the uninjured side invariably elicited a contralateral reflex response from the reinnervated muscles, while stimulation of the CP nerve on the injured side either failed to produce a response or produced a very weak reflex response from the control muscles. Reflexes recorded from the reinnervated muscles by stimulation of the tibial and sural branches of the uninjured sciatic nerve were 3-7 times greater than those recorded from the uninjured side or in normal animals. The reflex responses obtained from reinnervated muscles of animals with nerve injury in adulthood were similar to those obtained from control, unoperated adult rats. These results indicate that sciatic nerve injury during a critical development period leads to a permanent enhancement of reflex responses from reinnervated fast flexor muscles not seen after similar injury in adults.     

Comparative Efficacy of Once-Daily Umeclidinium/Vilanterol and Tiotropium/Olodaterol Therapy in Symptomatic Chronic Obstructive Pulmonary Disease: A Randomized Study

Introduction

We report the results of the first direct comparison of the once-daily fixed-dose long-acting muscarinic antagonist/long-acting β₂-agonist (LAMA/LABA) combinations umeclidinium/vilanterol (UMEC/VI) and tiotropium/olodaterol (TIO/OLO) in patients with COPD.

Methods

This was a randomized, two-period crossover open-label study in symptomatic patients with COPD [age 40 years or older, postbronchodilator forced expiratory volume in 1 s (FEV₁) of 70% or less and 50% or more of predicted normal values, and modified Medical Research Council Dyspnoea Scale score of 2 or greater] not receiving inhaled corticosteroid therapy. Patients were randomized to receive UMEC/VI (62.5/25 µg once daily) via a multidose dry powder inhaler (ELLIPTA) followed by TIO/OLO (5/5 µg once daily) via a soft mist inhaler (Respimat), each for 8 weeks with an interim 3-week washout or vice versa. The primary end point was the change from baseline in trough FEV₁ at week 8 with a noninferiority margin of ? 50 mL in the per-protocol (PP) population. The incidence of adverse events was also assessed.

Results

In total, 236 patients (mean age 64.4 years, 60% male) were included in the intent-to-treat population and 227 were included in the PP population. UMEC/VI treatment was noninferior in the PP population and superior in the intent-to-treat population to TIO/OLO treatment with regard to trough FEV₁ at week 8 [FEV₁ change from baseline 180 mL vs 128 mL; difference 52 mL (95% confidence interval 28–77 mL); p < 0.001]. Patients receiving UMEC/VI had twofold increased odds of experiencing a clinically meaningful increase (100 mL or more) from baseline in trough FEV₁ at week 8 compared with patients receiving TIO/OLO (odds ratio 2.05; 95% confidence interval 1.34–3.14). Adverse events occurred in 25% of patients in the UMEC/VI group and in 31% of patients in the TIO/OLO group.

Conclusion

In this first direct comparison of two once-daily fixed-dose LAMA/LABA combinations, superiority was observed for the primary end point of trough FEV₁ at week 8 with UMEC/VI compared with TIO/OLO in patients with symptomatic COPD. Both treatments had similar safety profiles. These findings confirm the results of previous indirect LAMA/LABA comparisons, and show that an efficacy gradient exists within the LAMA/LABA class.

Trial Registration

ClinicalTrials.gov identifier NCT02799784.

Funding

GlaxoSmithKline.