A New Method to Detect Directly in Culture Cell Surface Membrane Immunoglobulins

Abstract

An ELISA assay is described for the measurement of the smIgG. The method is based on the detection of cell-smIgG directly on the same microplate used for the culture. The cells, preincubated at 37°C for one hour, were cultured in the presence of S-ConA and serum-free medium for two days. Using this strategy, the background noise due to non specific adsorbtion of IgG to plastic wells and cytophilic antibodies was eliminated. The cells in the presence of S-ConA and serum-free medium adhered to the plastic wells, and the cell-smIgG were detected using an anti-human IgG covalently linked to alkaline phosphatase or its F(ab')₂ fragment. The possibility of measuring the modulation of the expression of the cell-smIgG without any additional manipulation is stressed.     

Chimeric antigen receptor-modified human regulatory T cells that constitutively express IL-10 maintain their phenotype and are potently suppressive

Clinical trials of Treg therapy in transplantation are currently entering phases IIa and IIb, with the majority of these employing polyclonal Treg populations that harbor a broad specificity. Enhancing Treg specificity is possible with the use of chimeric antigen receptors (CARs), which can be customized to respond to a specific human leukocyte antigen (HLA). In this study, we build on our previous work in the development of HLA-A2 CAR-Tregs by further equipping cells with the constitutive expression of interleukin 10 (IL-10) and an imaging reporter as additional payloads. Cells were engineered to express combinations of these domains and assessed for phenotype and function. Cells expressing the full construct maintained a stable phenotype after transduction, were specifically activated by HLA-A2, and suppressed alloresponses potently. The addition of IL-10 provided an additional advantage to suppressive capacity. This study therefore provides an important proof-of-principle for this cell engineering approach for next-generation Treg therapy in transplantation.     

Well-Being and Perceived Quality of Life in Elderly People Displaced After the Earthquake in L’Aquila,Italy

On 6 April 2009, the city of L’Aquila was hit by a violent earthquake that destroyed almost all of its medieval centre, and the surviving inhabitants were evacuated and relocated in temporary quarters or undamaged homes. The aim of this study was to investigate the perceived quality of life of the elderly population 3 years after the earthquake in relation to the social and logistic issues of new housing. The study was carried out between October 2011 and March 2012, and involved 571 subjects aged over 65 years living in the municipality of L’Aquila. The interviews took place in the surgeries of general practitioners and the city’s Department of Prevention and Vaccination in the anti-influenza immunisation period. The instrument used was a 36-item questionnaire with closed, multiple choice answers divided into the following sections: demographics, everyday activities, health and perceived health, and the quality of life in the city. The results show that, 3 years after the earthquake, the elderly population living in the new towns and temporary housing of L’Aquila have a worse perception of their quality of life than the others. They feel a certain social isolation and wish to live elsewhere. Governments faced with the problems arising from a natural calamity should take into account all of the elements making up a good quality of life and, before making choices whose impact cannot be changed, consider both their immediate and long-term social consequences.     

Mechanism of Resistance to an Antitubercular 2-Thiopyridine Derivative That Is Also Active against Burkholderia cenocepacia

The discovery of new compounds that are able to inhibit the growth of Burkholderia cenocepacia is of primary importance for cystic fibrosis patients. Here, the mechanism of resistance to a new pyridine derivative already shown to be effective against Mycobacterium tuberculosis and to have good activity toward B. cenocepacia was investigated. Increased expression of a resistance-nodulation-cell division (RND) efflux system was detected in the resistant mutants, thus confirming their important roles in B. cenocepacia antibiotic resistance.     

Cognitive processing and body balance in elderly subjects with vestibular dysfunction

Abnormal body balance and cognitive dysfunction may develop in elderly patients with chronic vestibular dysfunction.AimTo evaluate the relationship between cognitive processing and body balance in elderly patients with chronic peripheral vestibular disease. Type of Study: Cross-sectional.MethodsSeventy-six patients (≥ 60 years) with chronic peripheral vestibular dysfunction and dizziness for more than three months were enrolled. The tests for investigating body balance were: the Berg Balance Scale (BBS), Dynamic Gait Index (DGI), Timed Up and Go Test (TUGT) Timed Up and Go Test modified (TUGTm); the Mini Mental State Examination (MMSE), Test Clock (RT,) and Verbal Fluency Test (VF) were applied for assessing cognition.ResultsThe mean age was 69.03 years (SD=6.21 years); most were female (82.9%). There was a significant negative correlation between the MMSE and the TUGT (ρ=-0.312; p=0.01), MMSE and TUGTm (ρ=-0.306; p=0.01), FV and TUGT (ρ=-0.346; p=0.01), and FV and TUGTm (ρ=-0.536; p=0.01); there was a significant positive correlation between the TR and BBS (ρ=0.343; p=0.01), TR and DGI (ρ=0.298; p=0.01), FV and BBS (ρ=0.299; p=0.01), and FV and DGI (ρ=0.306; p=0.01).ConclusionElderly patients with chronic peripheral vestibular disease and worse performance in body balance tests have functional impairment in cognitive skills.     

A complex ventricular septal defect following transcatheter aortic valve implantation evaluated by 3D transthoracic echocardiography

Transcatheter aortic valve implantation (TAVI) has revolutionized the treatment of elderly patients with symptomatic severe aortic valve stenosis. Among the possible TAVI complications, a rare one is the annular/left ventricular outflow tract rupture. We report a rare case of a late complex ventricular septal defect (VSD) following TAVI with a balloon‐expandable prosthesis, conservatively managed. Our case demonstrates the role of 3D transthoracic echocardiography (3DTTE) in the accurate diagnosis of this TAVI complication and suggests that, in some cases, it can be used as an alternative to other diagnostic tools, such as transesophageal echocardiography, cardiac catheterization, and computed tomography.