Identification of new and unusual rev and nef transcripts expressed by an HIV type 1 primary isolate

We analyzed RNA splice site usage in three HIV-1 subtype B primary isolates through reverse transcriptase polymerase chain reaction (RT-PCR) amplification of spliced RNAs using a fluorescently labeled primer, with computerized size determination and quantification of PCR products, which were also identified by clone sequencing. In one isolate, P2149-3, unusual and unreported spliced transcripts were detected. This isolate preferentially used for rev RNA generation a 3' splice site (3'ss) located five nucleotides upstream of A4a, previously identified only in a T cell line-adapted virus and in a group O isolate, and designated A4d. P2149-3 also used an unreported 3'ss for rev RNA generation, designated A4h, located 20 nucleotides upstream of 3'ss A4c. Additionally, unusual nef RNAs using 3'ss A5a and A7a and with exon composition 1.3.7 were identified. The identification of several unusual and unreported spliced transcripts in an HIV-1 primary isolate suggests a greater diversity of splice site usage in HIV-1 than previously appreciated.We thank the personnel at the Genomic Unit of Centro Nacional de Microbiología, Instituto de Salud Carlos III, for technical assistance in sequencing and GeneMapper analyses. This work was funded by Ministerio de Economía y Competitividad (Spain), Plan Nacional de I+D+I, through grants SAF2007-61688 and SAF2010-2096. Sequences of PCR clones derived from P2149-3 DS transcripts have been deposited in GenBank under accession numbers JF808039–JF808078

Floor and Ceiling Effects, Time to Completion, and Question Burden of PROMIS CAT Domains Among Shoulder and Knee Patients Undergoing Nonoperative and Operative Treatment

The Patient-Reported Outcomes Measurement Information System (PROMIS) computer adaptive tests (CATs) have emerged as an efficient technique for measuring patient-reported outcomes among orthopaedic patients. The purpose of this study was to investigate the floor and ceiling (F/C) effects, time to completion (TTC), and question burden of PROMIS CATs administered to patients presenting to a shoulder and sports medicine orthopaedic clinic. Methods: Patients prospectively completed PROMIS CATs including the physical function (PROMIS-PF) or upper-extremity function (PROMIS-UE), pain interference (PROMIS-PI), and depression (PROMIS-D) domains at their initial encounter and were retrospectively included in this study. Adult patients indicating a single problem involving either the shoulder or knee were included. Patients were also grouped as either preoperative or nonoperative. F/C effects were defined as the proportion of respondents scoring the highest (ceiling) or lowest (floor) possible score across a given domain. Results: Included were 2,952 patients (average age, 51.0 ± 16.9 years). The PROMIS-UE, PROMIS-PF, and PROMIS-PI demonstrated negligible F/C effects across all shoulder and knee patients (\u3c2%). The PROMIS-D displayed moderate to significant floor effects (13.9% to 18.9%) and a 0% ceiling effect in all main patient groups. The mean TTC and mean question burden of the PROMIS-UE, PROMIS-PF, and PROMIS-PI ranged from 45.3 to 54.4 seconds and 4.1 to 4.9 questions for all patient groups, while the PROMIS-D exhibited a TTC ranging from 20.9 to 38.6 seconds for all groups and a question burden that ranged from 6.2 to 6.7 questions. Conclusions: The PROMIS-PF, PROMIS-UE, and PROMIS-PI demonstrated favorable F/C effects, TTC, and question burden among both nonoperative and preoperative patients. These findings justify consideration of the PROMIS-PF, PROMIS-UE, and PROMIS-PI for clinical and research applications involving shoulder and knee sports medicine patients. Additionally, we found moderate to significant floor effects for the PROMIS-D in all patient groups, which may be multifactorial in nature and may not be unexpected in patients with an isolated joint concern. Clinical Relevance: This study highlights the psychometric properties of PROMIS CAT forms for knee and shoulder patients. Understanding these basic properties is important in considering the adoption of PROMIS CAT forms for patients with musculoskeletal conditions

Regulation of death receptor signaling by the autophagy protein TP53INP2

TP53INP2 positively regulates autophagy by binding to Atg8 proteins. Here, we uncover a novel role of TP53INP2 in death‐receptor signaling. TP53INP2 sensitizes cells to apoptosis induced by death receptor ligands. In keeping with this, TP53INP2 deficiency in cultured cells or mouse livers protects against death receptor‐induced apoptosis. TP53INP2 binds caspase‐8 and the ubiquitin ligase TRAF6, thereby promoting the ubiquitination and activation of caspase‐8 by TRAF6. We have defined a TRAF6‐interacting motif (TIM) and a ubiquitin‐interacting motif in TP53INP2, enabling it to function as a scaffold bridging already ubiquitinated caspase‐8 to TRAF6 for further polyubiquitination of caspase‐8. Mutations of key TIM residues in TP53INP2 abrogate its interaction with TRAF6 and caspase‐8, and subsequently reduce levels of death receptor‐induced apoptosis. A screen of cancer cell lines showed that those with higher protein levels of TP53INP2 are more prone to TRAIL‐induced apoptosis, making TP53INP2 a potential predictive marker of cancer cell responsiveness to TRAIL treatment. These findings uncover a novel mechanism for the regulation of caspase‐8 ubiquitination and reveal TP53INP2 as an important regulator of the death receptor pathway

Ferric carboxymaltose with or without erythropoietin for the prevention of red-cell transfusions in the perioperative period of osteoporotic hip fractures: a randomized contolled trial. The PAHFRAC-01 project

Background: Around one third to one half of patients with hip fractures require red-cell pack transfusion. The increasing incidence of hip fracture has also raised the need for this scarce resource. Additionally, red-cell pack transfusions are not without complications which may involve excessive morbidity and mortality. This makes it necessary to develop blood-saving strategies. Our objective was to assess safety, efficacy, and cost-effictveness of combined treatment of i.v. ferric carboxymaltose and erythropoietin (EPOFE arm) versus i.v. ferric carboxymaltose (FE arm) versus a placebo (PLACEBO arm) in reducing the percentage of patients who receive blood transfusions, as well as mortality in the perioperative period of hip fracture intervention. Methods/Design: Multicentric, phase III, randomized, controlled, double blinded, parallel groups clinical trial. Patients > 65 years admitted to hospital with a hip fracture will be eligible to participate. Patients will be treated with either a single dosage of i.v. ferric carboxymaltose of 1 g and subcutaneous erythropoietin (40.000 IU), or i.v. ferric carboxymaltose and subcutaneous placebo, or i.v. placebo and subcutaneous placebo. Follow-up will be performed until 60 days after discharge, assessing transfusion needs, morbidity, mortality, safety, costs, and health-related quality of life. Intention to treat, as well as per protocol, and incremental cost-effectiveness analysis will be performed. The number of recruited patients per arm is set at 102, a total of 306 patients. Discussion: We think that this trial will contribute to the knowledge about the safety and efficacy of ferric carboxymaltose with/without erythropoietin in preventing red-cell pack transfusions in patients with hip fracture. ClinicalTrials.gov identifier: NCT01154491

Timing of surgery for hip fracture and in-hospital mortality: a retrospective population-based cohort study in the Spanish National Health System

<p>Abstract</p> <p>Background</p> <p>While the benefits or otherwise of early hip fracture repair is a long-running controversy with studies showing contradictory results, this practice is being adopted as a quality indicator in several health care organizations. The aim of this study is to analyze the association between early hip fracture repair and in-hospital mortality in elderly people attending public hospitals in the Spanish National Health System and, additionally, to explore factors associated with the decision to perform early hip fracture repair.</p> <p>Methods</p> <p>A cohort of 56,500 patients of 60-years-old and over, hospitalized for hip fracture during the period 2002 to 2005 in all the public hospitals in 8 Spanish regions, were followed up using administrative databases to identify the time to surgical repair and in-hospital mortality. We used a multivariate logistic regression model to analyze the relationship between the timing of surgery (< 2 days from admission) and in-hospital mortality, controlling for several confounding factors.</p> <p>Results</p> <p>Early surgery was performed on 25% of the patients. In the unadjusted analysis early surgery showed an absolute difference in risk of mortality of 0.57 (from 4.42% to 3.85%). However, patients undergoing delayed surgery were older and had higher comorbidity and severity of illness. Timeliness for surgery was not found to be related to in-hospital mortality once confounding factors such as age, sex, chronic comorbidities as well as the severity of illness were controlled for in the multivariate analysis.</p> <p>Conclusions</p> <p>Older age, male gender, higher chronic comorbidity and higher severity measured by the Risk Mortality Index were associated with higher mortality, but the time to surgery was not.</p

p38α Mitogen-activated Protein Kinase Sensitizes Cells to Apoptosis Induced by Different Stimuli

p38α mitogen-activated protein (MAP) kinase is a broadly expressed signaling molecule that participates in the regulation of cellular responses to stress as well as in the control of proliferation and survival of many cell types. We have used cell lines derived from p38α knockout mice to study the role of this signaling pathway in the regulation of apoptosis. Here, we show that cardiomyocytes and fibroblasts lacking p38α are more resistant to apoptosis induced by different stimuli. The reduced apoptosis of p38α-deficient cells correlates with decreased expression of the mitochondrial proapoptotic protein Bax and the apoptosis-inducing receptor Fas/CD-95. Cells lacking p38α also have increased extracellular signal-regulated kinase (ERKs) MAP kinase activity, and the up-regulation of this survival pathway seems to be at least partially responsible for the reduced levels of apoptosis in the absence of p38α. Phosphorylation of the transcription factor STAT3 on Ser-727, mediated by the extracellular signal-regulated kinase MAP kinase pathway, may contribute to the decrease in both Bax and Fas expression in p38α-/- cells. Thus, p38α seems to sensitize cells to apoptosis via both up-regulation of proapoptotic proteins and down-regulation of survival pathways