Emergency Veno-Arterial Extracorporeal Membrane Oxygenation for Pericardial Decompression Syndrome.

Background: Pericardiocentesis is a therapeutic lifesaving intervention for patients presenting with cardiogenic shock due to pericardial effusion with signs of tamponade. Pericardial decompression syndrome (PDS) is a rare fatal complication that may occur after pericardiocentesis. Case Presentation. We report a case of a patient with idiopathic primary pulmonary hypertension who presented with massive pericardial effusion complicated with rapid hemodynamic and respiratory deterioration. Gradual therapeutic pericardiocentesis was done but progressive circulatory collapse occurred. Emergent veno-arterial extracorporeal membrane oxygenation (VA-ECMO) was applied. Echocardiography revealed severe right ventricle failure. Unfortunately, the patient developed acute progressive thrombocytopenia and bilaterally diffuse subarachnoid hemorrhage after 4 days of ECMO support. Conclusions: Therapeutic pericardiocentesis can be occasionally fatal in cases of significant pulmonary hypertension with massive pericardial effusion when complicated by pericardial decompression syndrome. Acute significant thrombocytopenia may occur with VA-ECMO support resulting in fatal bleeding.

Identification of Key Coagulation Activity Determining Elements in Canine Factor VIII.

It is well known that canine factor VIII (cFVIII) has a higher specific activity than does human FVIII (hFVIII), and it has been previously demonstrated that cFVIII light chain is able to enhance hFVIII activity. The goal of this study was to first determine which amino acids in cFVIII light chain were responsible for enhancing hFVIII activity, and second to use these amino acids to develop a hFVIII variant with enhanced functional activity. We systemically screened segments of cFVIII light chain by testing an array of human-canine light chain hybrids and found that canine amino acids 1857-2147 were key to this enhancement. Each canine amino acid within this span was screened individually using a negative selection method, which led to the identification of 12 aa (JF12) in the FVIII light chain that could enhance activity. Substitution of the corresponding 12 aa into hFVIII (hFVIIIJF12BDD) elevated the specific activity profile in vitro. Furthermore, hFVIIIJF12BDD expressed an in vivo-displayed increased coagulation activity compared to wild-type, while maintaining normal secretion efficiency. In conclusion, we identified the amino acids in cFVIII that are the key determinants for higher specific activity and may be the basis for future development of therapeutic treatments for hemophilia A.

Clinical and radiological outcomes of acetabular revision with the Delta Revision TT cup.

INTRODUCTION:: This study evaluates mid-term results of acetabular revision using a hemispherical acetabular cup in Trabecular Titanium with a cage construct. METHODS:: We reviewed 36 acetabular revisions performed with the Delta Revision TT cup in 34 patients (mean age = 75, range: 45-92 years). Acetabular defect types (Paprosky classification) included (1) 2B ( n = 5), (2) 2C ( n = 7), (3) 3A ( n = 15), and (4) 3B ( n = 9). Morcellised bone allografts were used in 24 cases, and synthetic bone used in 11. Outcomes were evaluated using the Harris Hip Score (HHS), and the Verbal Rating Scale (VRS) for pain measurement. X-ray visualisation of cup position was used to discern signs of mobilisation and bone graft incorporation. Survivorship at post-revision follow-up (mean = 39.8; range 12-91.5 months) was calculated. RESULTS:: HHS increased from 40.5 to 87 ( p < 0.01). 68% of cases were pain free; by comparison, 32% had an average VRS score of 1.9 (range 1-3). The average cup inclination angle was 40.8° (30-52°) postoperatively, compared with 41.2° (30-52°) at follow-up; there were no signs of loosening or mobilisation. Centre of rotation was fully restored in 21 (58.3%) hips. According to Gie classification; bone graft incorporation grades were (1) 3 ( n = 21), (2) 2 ( n = 12) and (3) 1 ( n = 2). The survival rate was 100% for aseptic loosening and 91.7% for any cause of revision. CONCLUSIONS:: The Delta Revision TT cup promises good clinical and radiographic results at short- to mid-term follow-up, with high rates of survival rate and bone integration.

Efficient production of dual recombinant adeno-associated viral vectors for factor VIII delivery.

Recombinant adeno-associated viral (rAAV) vectors have gained attention for human gene therapy because of their high safety and clinical efficacy profile. For factor VIII gene delivery, splitting the coding region between two AAV vectors remains a viable strategy to avoid the packaging capacity limitation (∼5.0 kb). However, it is time-consuming and labor-intensive to produce two rAAV vectors in separate batches. Here we demonstrated successful production of dual rAAV vectors for hemophilia A gene therapy in a single preparation. When the AAV vector plasmids carrying the human factor VIII heavy chain (hHC) and the light chain (hLC) expression cassettes were cotransfected into 293 cells along with the AAV rep&cap and mini-adenovirus helper plasmids, both rAAV-hHC and rAAV-hLC were produced at the desired ratio and in high titer. Interestingly, the rAAV-hHC vectors always yielded higher titers than rAAV-hLC vectors as a result of more efficient replication of rAAV-hHC genomes. The resulting vectors were effective in transducing the tissue culture cells in vitro. When these vectors were administered to hemophilia A mice, factor VIII was detected in the mouse plasma by both the activated partial thromboplastin time assay and enzyme-linked immunosorbent assay. The functional activity as well as the antigen levels of secreted factor VIII were similar to those of vectors produced by the traditional method. The dual-vector production method has been successfully extended to both AAV2 and AAV8 serotypes. In conclusion, cotransfection of vector plasmids presents an efficient method for producing dual or multiple AAV vectors at significantly reduced cost and labor.

Resolution of inflammation in murine autoimmune arthritis is disrupted by cyclooxygenase-2 inhibition and restored by prostaglandin E2-mediated lipoxin A4 production.

Acute inflammation follows defined phases of induction, inflammation and resolution, and resolution occurs by an active process that requires cyclooxygenase-2 (COX-2) activity. This study aims to address whether this paradigm extends to recognized model of chronic inflammation. We demonstrated that murine collagen-induced arthritis follows a similar sequential course. Interestingly, COX-2 and its metabolite, the presumably proinflammatory PGE(2), are present in the joints during resolution, and blocking COX-2 activity and PGE(2) production within this period perpetuated, instead of attenuated, inflammation. Repletion with PGE(2) analogs restored homeostasis, and this function is mediated by the proresolving lipoxygenase metabolite, lipoxin A(4), a potent stop signal. Thus, the study provided in vivo evidence for a natural, endogenous link between the cyclooxygenase-lipoxygenase pathways and showed that PGE(2) serves as a feedback inhibitor essential for limiting chronic inflammation in autoimmune arthritis. These findings may explain the enigma regarding why COX-2 inhibitors are palliative rather than curative in humans, because blocking resolution may mitigate the benefit of preventing induction.

Synaptotagmin 2 couples mucin granule exocytosis to Ca2+ signaling from endoplasmic reticulum.

Synaptotagmin 2 (Syt2) functions as a low affinity, fast exocytic Ca(2+) sensor in neurons, where it is activated by Ca(2+) influx through voltage-gated channels. Targeted insertion of lacZ into the mouse syt2 locus reveals expression in mucin-secreting goblet cells of the airways. In these cells, rapid Ca(2+) entry from the extracellular medium does not contribute significantly to stimulated secretion (Davis, C. W., and Dickey, B. F. (2008) Annu. Rev. Physiol. 70, 487-512). Nonetheless, Syt2(-/-) mice show a severe defect in acute agonist-stimulated airway mucin secretion, and Syt2(+/-) mice show a partial defect. In contrast to Munc13-2(-/-) mice (Zhu, Y., Ehre, C., Abdullah, L. H., Sheehan, J. K., Roy, M., Evans, C. M., Dickey, B. F., and Davis, C. W. (2008) J. Physiol. (Lond.) 586, 1977-1992), Syt2(-/-) mice show no spontaneous mucin accumulation, consistent with the inhibitory action of Syt2 at resting cytoplasmic Ca(2+) in neurons. In human airway goblet cells, inositol trisphosphate receptors are found in rough endoplasmic reticulum that closely invests apical mucin granules, consistent with the known dependence of exocytic Ca(2+) signaling on intracellular stores in these cells. Hence, Syt2 can serve as an exocytic sensor for diverse Ca(2+) signaling systems, and its levels are limiting for stimulated secretory function in airway goblet cells.

Efeitos da administração sistêmica de corticosteróides sobre propriedades mecânicas do diafragma de coelhos / Effects of corticosteroid systemic administration on mechanical properties of rabbits diaphragm muscle

Corticosteróides sistêmicos em altas doses podem causar miopatia metabólica. O objetivo deste estudo foi avaliar, por meio de ensaios de tração, os efeitos da miopatia induzida por corticosteróides nas propriedades mecânicas do músculo diafragma de coelhos...

High doses of systemic corticosteroids might promote metabolic myopathy. The aim of this study was to assess, by means of traction assays, the effects of myopathy induced by systemic corticosteroids on mechanical features of the diaphragm muscle of rabbits...