The microRNA miR-31 inhibits CD8+ T cell function in chronic viral infection.

During infection, antigen-specific T cells undergo tightly regulated developmental transitions controlled by transcriptional and post-transcriptional regulation of gene expression. We found that the microRNA miR-31 was strongly induced by activation of the T cell antigen receptor (TCR) in a pathway involving calcium and activation of the transcription factor NFAT. During chronic infection with lymphocytic choriomeningitis virus (LCMV) clone 13, miR-31-deficent mice recovered from clinical disease, while wild-type mice continued to show signs of disease. This disease phenotype was explained by the presence of larger numbers of cytokine-secreting LCMV-specific CD8+ T cells in miR-31-deficent mice than in wild-type mice. Mechanistically, miR-31 increased the sensitivity of T cells to type I interferons, which interfered with effector T cell function and increased the expression of several proteins related to T cell dysfunction during chronic infection. These studies identify miR-31 as an important regulator of T cell exhaustion in chronic infection.

An analysis of the recording inpatients in Irish hospitals of tobacco use among.

Smoking is the largest avoidable cause of premature mortality in the world. Hospital admission is an opportunity to identify and help smokers quit. This study aimed to determine the level of recording of tobacco use (current and past) in Irish hospitals. Information on inpatient discharges with a tobacco use diagnosis was extracted from HIPE. In 2011, a quarter (n=84, 679) of discharges had a recording of tobacco use, which were more common among males (29% (n=50,161) male v. 20% (n=30,162) female), among medical patients (29% (n=54,375) medical v. 20% (n=30,162) other) and was highest among those aged 55-59 years (30.6%; n=7,885). SLAN 2007 reported that 48% of adults had smoked at some point in their lives. This study would suggest an under- reporting of tobacco use among hospital inpatients. Efforts should be made to record smoking status at hospital admission, and to improve the quality of the HIPE coding of tobacco use.

An audit of smoking prevalence and awareness of HSE smoking cessation services among HSE staff.

This audit estimated smoking prevalence and awareness of quit services among Health Service Executive (HSE) staff. A questionnaire posted to a random sample of 1,064 staff received a 71% response rate. Staff smoking prevalence was 15.0% overall, and 4.4% among Medical/Dental staff. Front-line-healthcare staff were less likely to smoke than other staff categories (adjusted OR 0.38, p < 0.001). Only 63.6% of staff were aware of HSE quit services. Targeted interventions are required to help staff to quit smoking and to boost awareness of quit services.

Asa Issie, Aramis and the origin of Australopithecus.

The origin of Australopithecus, the genus widely interpreted as ancestral to Homo, is a central problem in human evolutionary studies. Australopithecus species differ markedly from extant African apes and candidate ancestral hominids such as Ardipithecus, Orrorin and Sahelanthropus. The earliest described Australopithecus species is Au. anamensis, the probable chronospecies ancestor of Au. afarensis. Here we describe newly discovered fossils from the Middle Awash study area that extend the known Au. anamensis range into northeastern Ethiopia. The new fossils are from chronometrically controlled stratigraphic sequences and date to about 4.1-4.2 million years ago. They include diagnostic craniodental remains, the largest hominid canine yet recovered, and the earliest Australopithecus femur. These new fossils are sampled from a woodland context. Temporal and anatomical intermediacy between Ar. ramidus and Au. afarensis suggest a relatively rapid shift from Ardipithecus to Australopithecus in this region of Africa, involving either replacement or accelerated phyletic evolution.

Safety switch optimization enhances antibody-mediated elimination of CAR T cells.

Activation of a conditional safety switch has the potential to reverse serious toxicities arising from the administration of engineered cellular therapies, including chimeric antigen receptor (CAR) T cells. The functionally inert, non-immunogenic cell surface marker derived from human epidermal growth factor receptor (EGFRt) is a promising safety switch that has been used in multiple clinical constructs and can be targeted by cetuximab, a clinically available monoclonal antibody. However, this approach requires high and persistent cell surface expression of EGFRt to ensure that antibody-mediated depletion of engineered cells is rapid and complete. Here we show that incorporating a short juxtamembrane sequence into the EGFRt polypeptide enhances its expression on the surface of T cells and their susceptibility to antibody-dependent cellular cytotoxicity (ADCC). Incorporating this optimized variant (EGFRopt) into bicistronic and tricistronic CAR designs results in more rapid in vivo elimination of CAR T cells and robust termination of their effector activity compared to EGFRt. These studies establish EGFRopt as a superior safety switch for the development of next-generation cell-based therapeutics.

Admissions and costs to acute hospitals resulting from road traffic crashes, 2005-2009.

Road traffic crashes (RTCs) remain a leading cause of death and injury. The aim of this study was to explore the use of hospital data as a source of RTC-related injury data in Ireland, as current systems are believed to under-estimate the burden. Information on inpatient discharges for years 2005-2009, admitted with RTC-related injuries were extracted from HIPE. There were 14,861 discharges; 9,661 (65.0%) were male, with an average age of 33 years. The median length of stay was two days. The most common diagnosis was head injury (n = 4,644; 31.2%). The average inpatient hospital cost was Euro 6,395 per discharge. 1,498 (10.1%) were admitted to intensive care units. This study has identified 3.5 times more serious injuries (14,861) than identified in the Road Safety Authority (RSA) statistics (4,263) indicating that the extent of road injuries is greater than previously estimated. Hospital data could be used annually in conjunction with RSA and other data; ideally the data should be linked.

Determining the cost of genital warts: a study from Ireland.

OBJECTIVES: To determine the average cost of a case of genital warts, for both males and females, with a view to informing the current debate as to which Human papillomavirus vaccine would have maximum cost-effectiveness in the Irish population. METHODS: Contact time between patients and healthcare professionals was prospectively measured at five genitourinary medicine clinics in the south-west of Ireland, over a period of 3 weeks. By identifying all those with genital warts, it was possible to calculate the proportion of total time taken by patients with this condition, and from this to calculate a cost per incident case, by gender. RESULTS: A total of 25.5% of attendances were for genital warts, and these patients used 26.2% of total clinic time (CI 25.4 to 27.0%). The average cost calculated for genital warts was 335 euros per incident case, and by gender 300 euros per male case and 366 euros per female case. CONCLUSIONS: There are considerable costs associated with the treatment of genital warts, with female cases representing a higher cost than males. By vaccinating with the quadrivalent HPV vaccine, there are significant savings to be made.

Not miR-ly small RNAs: big potential for microRNAs in therapy.

RNA interference (RNAi) describes a set of natural processes in which genes are silenced by small RNAs. RNAi has been widely used as an experimental tool that has recently become the focus of drug development efforts to treat a variety of diseases and disorders. Like all molecular therapies, in vivo delivery is the major hurdle to realizing therapeutic RNAi. Several strategies have been developed that increase small RNA half-life in the blood, facilitate transduction across biological membranes, and mediate cell-specific delivery. Importantly, these strategies permit targeting of mRNAs as well as microRNAs (miRNAs), a class of small RNAs encoded in the genome. miRNAs are required for multiple developmental and cellular processes. Dysfunction of miRNAs can result in a host of pathologies, suggesting that miRNAs are potential targets of therapy. Recent studies of miRNA function in immune-specific pathways indicate that specific miRNAs might be exploited as therapeutic targets to treat immune disorders, including autoimmunity, allergy, and hematopoietic cancers.

Alcohol levels in killed drivers and pedestrians on Irish roads 2003-2005: a national study.

No official data are provided in Ireland to indicate what proportion of the deaths on Irish roads have alcohol as a contributory factor. The aim of this study was to identify the blood alcohol concentration (BAC) in fatally injured drivers and pedestrians in Ireland. An Garda Síochána (The Irish police) gather data on all fatal road crashes and individual paper files are kept on each crash. The authors examined all such files for deaths in 2003-2005. Of the 611 drivers fatally injured, 184 (30.1%) were over the BAC legal limit (80 mg/100 ml). BACs were available for only 397 (64.9%) of drivers. Of the 397 drivers who had their BACs recorded, 184 (46.3%) had a BAC over the legal limit of 80 mg/100 ml and 220 (55.4%) had BACs 20 mg/100 ml or higher. Fatally injured drivers with BACs 20 mg/100 ml or greater were more likely to be male (88.6%/o, p<0.01). Alcohol-related crashes were more likely to occur on week end nights. Pedestrian alcohol consumption was considered to be a contributory factor in 50 (24.4%) of the pedestrian deaths with 22 (10.7%) of the pedestrians having alcohol levels exceeding 240 mgl/100 ml. This study confirms that alcohol is a significant factor in road deaths. Further targeted action including a reduction in the legal limit is required.

B cells engineered to express pathogen-specific antibodies protect against infection.

Effective vaccines inducing lifelong protection against many important infections such as respiratory syncytial virus (RSV), HIV, influenza virus, and Epstein-Barr virus (EBV) are not yet available despite decades of research. As an alternative to a protective vaccine, we developed a genetic engineering strategy in which CRISPR-Cas9 was used to replace endogenously encoded antibodies with antibodies targeting RSV, HIV, influenza virus, or EBV in primary human B cells. The engineered antibodies were expressed efficiently in primary B cells under the control of endogenous regulatory elements, which maintained normal antibody expression and secretion. Using engineered mouse B cells, we demonstrated that a single transfer of B cells engineered to express an antibody against RSV resulted in potent and durable protection against RSV infection in RAG1-deficient mice. This approach offers the opportunity to achieve sterilizing immunity against pathogens for which traditional vaccination has failed to induce or maintain protective antibody responses.

Association of intra-operative metaphyseal fractures with prosthesis size during hemiarthroplasty of the hip.

PURPOSE: To study the possible causes of intra-operative metaphyseal fractures in elderly patients undergoing hemiarthroplasty for displaced intracapsular femoral neck fracture. METHODS: 36 men and 228 women aged 61 to 89 years with 273 displaced femoral neck fractures underwent hemiarthroplasty using a hydroxyapatite ceramic-coated Furlong bipolar prosthesis. Anteroposterior and lateral radiographs were taken immediately after surgery to evaluate the presence and type of any intra-operative fractures (classified according to the Vancouver system) and their effect on stem stability or osseointegration. Pain and clinical outcomes were evaluated using a visual analogue scale and the Harris Hip Score. RESULTS: Regarding the 273 surgeries for displaced femoral neck fracture, 28 (10%) were associated with intra-operative metaphyseal fracture (21 Vancouver type AL and 7 type AG). There was a correlation between intra-operative metaphyseal fractures and stem size. A size-9 stem was used in 64 surgeries without any fracture. A size-10 stem was used in 129 surgeries in which 11 (9%) sustained fractures, and a size-12 stem was used in 80 surgeries in which 17 (21%) sustained fractures. Postoperatively, 25 cases developed hip-related problems (thigh pain=14 and periprosthetic fractures=8) after 3 to 18 months. No patient whose metaphyseal fracture was fixed had hip problems. CONCLUSION: In elderly women with compromised bone quality, extra care is needed to achieve better fitting so as to avoid iatrogenic metaphyseal fractures. Under-sizing or cementing of the prosthesis is recommended when encountering difficulties.

The impact of legislation in Ireland on handheld mobile phone use by drivers.

Under the Road Traffic Act, 2006 handheld mobile phone use whilst driving is an offence liable to a fine and penalty points. The aim of this study was to determine whether there has been a change in driver behaviour following the introduction of this legislation. This study found that 2.3% of drivers were still using a handheld mobile phone.

The growth inhibition of human breast cancer cells by a novel synthetic progestin involves the induction of transforming growth factor beta.

Recent experimental work has identified a novel intracellular binding site for the synthetic progestin, Gestodene, that appears to be uniquely expressed in human breast cancer cells. Gestodene is shown here to inhibit the growth of human breast cancer cells in a dose-dependent fashion, but has no effect on endocrine-responsive human endometrial cancer cells. Gestodene induced a 90-fold increase in the secretion of transforming growth factor-beta (TGF-beta) by T47D human breast cancer cells. Other synthetic progestins had no effect, indicating that this induction is mediated by the novel Gestodene binding site and not by the conventional progesterone receptor. Furthermore, in four breast cancer cell lines, the extent of induction of TGF-beta correlated with intracellular levels of Gestodene binding site. No induction of TGF-beta was observed with the endometrial cancer line, HECl-B, which lacks the Gestodene binding site, but which expresses high levels of progesterone receptor. The inhibition of growth of T47D cells by Gestodene is partly reversible by a polyclonal antiserum to TGF-beta. These data indicate that the growth-inhibitory action of Gestodene may be mediated in part by an autocrine induction of TGF-beta.

Lymphocyte phenotype and function in pseudolymphoma associated with Sjögren's syndrome.

Lymph node (LNL) and salivary gland lymphocytes (SGL) from three patients with pseudolymphoma and primary Sjögren's syndrome (1(0)SS) were characterized with monoclonal antibodies to demonstrate (a) a predominance of T cells (greater than 80%) reactive with anti-T cell antibodies OKT4 (greater than 70%) and OKT8 (less than 20%); (b) a high prevalence of activation antigens (greater than 50% of cells reactive with antibody OKT10 and anti-Ia antibody); (c) polyclonal B cells (8-15% of all cells expressing kappa or lambda); and (d) a specific B cell subset defined by reactivity with antibody B532 that was not present in their peripheral blood. In vitro functional studies showed that both SGL and LNL provided T helper activity for immunoglobulin synthesis and that this activity could be abolished by treatment with antibody OKT4 plus complement. The SGL and LNL exhibited little natural killer, antibody-dependent cellular cytotoxicity, or cytotoxic T cell activity. Normal karyotype was observed in SGL, LNL, and peripheral blood lymphocytes (PBL) from these patients. These findings indicate that pseudolymphoma in 1(0)SS results from the infiltration of salivary glands and extraglandular tissues by nonneoplastic T helper cells. Monoclonal antibodies provide an important tool to distinguish pseudolymphoma from non-Hodgkins (B cell) lymphomas that have a markedly elevated incidence in 1(0)SS patients. Our finding of T helper cells in pseudolymphoma tissues supports the hypothesis that chronic stimulation of B cells by helper T cells leads to eventual escape of a malignant B cell clone.

Staged bilateral hip or knee arthroplasties.

PURPOSE: To compare the operating time, amount of blood transfused, length of hospital stay, and early complications (within 6 months) between 2-week staged bilateral arthroplasties and matched randomised controls undergoing unilateral arthroplasties. METHODS: From October 1992 to October 2000, 90 patients who underwent bilateral hip or knee arthroplasties with a 2-week interval were compared with matched randomised controls undergoing unilateral arthroplasties. A single surgeon performed all procedures. RESULTS: After the match-up process, 30 pairs of patients were included in the analysis. There were no significant differences in the operating times, amount of blood transfused, and early complication rates. The mean difference in length of hospital stay was significant (t=-3.552, df=29, p<0.001). CONCLUSION: Compared to staged procedures with an interval months apart, staged sequential arthroplasty with a 7- to 10-day interval during one hospital admission is more efficient, as it facilitates earlier rehabilitation without higher complication rates, and entails shorter hospital stays.

Antagonism of HIV-specific CD4+ T cells by C-terminal truncation of a minimum epitope.

Antagonism of T cell responses by variants of the cognate peptide is a potential mechanism of viral escape from immune responses and may play a role in the ability of HIV to evade immune control. We show here a rarely described mechanism of antagonism by a peptide shorter than the minimum length epitope for an HIV p24-specific CD4+ T cell clone. The shorter antagonist peptide-MHC complex bound the T cell receptor (TCR), albeit with lower affinity than the full-length agonist peptide. Prior work showing the crystal structure of the peptide-MHC complex revealed a unique glycine hinge near the C-terminus of the agonist peptide, allowing the generation of full-length antagonist peptide lacking the hinge. These results confirm the dependence of productive TCR engagement on residues spilling out from the C-terminus of the MHC binding groove and show that partial engagement of the TCR with a truncated, low-affinity ligand can result in T cell antagonism.

In situ programming of leukaemia-specific T cells using synthetic DNA nanocarriers.

An emerging approach for treating cancer involves programming patient-derived T cells with genes encoding disease-specific chimeric antigen receptors (CARs), so that they can combat tumour cells once they are reinfused. Although trials of this therapy have produced impressive results, the in vitro methods they require to generate large numbers of tumour-specific T cells are too elaborate for widespread application to treat cancer patients. Here, we describe a method to quickly program circulating T cells with tumour-recognizing capabilities, thus avoiding these complications. Specifically, we demonstrate that DNA-carrying nanoparticles can efficiently introduce leukaemia-targeting CAR genes into T-cell nuclei, thereby bringing about long-term disease remission. These polymer nanoparticles are easy to manufacture in a stable form, which simplifies storage and reduces cost. Our technology may therefore provide a practical, broadly applicable treatment that can generate anti-tumour immunity 'on demand' for oncologists in a variety of settings.

Biopolymers codelivering engineered T cells and STING agonists can eliminate heterogeneous tumors.

Therapies using T cells that are programmed to express chimeric antigen receptors (CAR T cells) consistently produce positive results in patients with hematologic malignancies. However, CAR T cell treatments are less effective in solid tumors for several reasons. First, lymphocytes do not efficiently target CAR T cells; second, solid tumors create an immunosuppressive microenvironment that inactivates T cell responses; and third, solid cancers are typified by phenotypic diversity and thus include cells that do not express proteins targeted by the engineered receptors, enabling the formation of escape variants that elude CAR T cell targeting. Here, we have tested implantable biopolymer devices that deliver CAR T cells directly to the surfaces of solid tumors, thereby exposing them to high concentrations of immune cells for a substantial time period. In immunocompetent orthotopic mouse models of pancreatic cancer and melanoma, we found that CAR T cells can migrate from biopolymer scaffolds and eradicate tumors more effectively than does systemic delivery of the same cells. We have also demonstrated that codelivery of stimulator of IFN genes (STING) agonists stimulates immune responses to eliminate tumor cells that are not recognized by the adoptively transferred lymphocytes. Thus, these devices may improve the effectiveness of CAR T cell therapy in solid tumors and help protect against the emergence of escape variants.

Reductions in tobacco smoke pollution and increases in support for smoke-free public places following the implementation of comprehensive smoke-free workplace legislation in the Republic of Ireland: findings from the ITC Ireland/UK Survey.

OBJECTIVE: To evaluate the psychosocial and behavioural impact of the first ever national level comprehensive workplace smoke-free law, implemented in Ireland in March 2004. DESIGN: Quasi-experimental prospective cohort survey: parallel cohort telephone surveys of national representative samples of adult smokers in Ireland (n = 769) and the UK (n = 416), surveyed before the law (December 2003 to January 2004) and 8-9 months after the law (December 2004 to January 2005). MAIN OUTCOME MEASURES: Respondents' reports of smoking in key public venues, support for total bans in those key venues, and behavioural changes due to the law. RESULTS: The Irish law led to dramatic declines in reported smoking in all venues, including workplaces (62% to 14%), restaurants (85% to 3%), and bars/pubs (98% to 5%). Support for total bans among Irish smokers increased in all venues, including workplaces (43% to 67%), restaurants (45% to 77%), and bars/pubs (13% to 46%). Overall, 83% of Irish smokers reported that the smoke-free law was a "good" or "very good" thing. The proportion of Irish homes with smoking bans also increased. Approximately 46% of Irish smokers reported that the law had made them more likely to quit. Among Irish smokers who had quit at post-legislation, 80% reported that the law had helped them quit and 88% reported that the law helped them stay quit. CONCLUSION: The Ireland smoke-free law stands as a positive example of how a population-level policy intervention can achieve its public health goals while achieving a high level of acceptance among smokers. These findings support initiatives in many countries toward implementing smoke-free legislation, particularly those who have ratified the Framework Convention on Tobacco Control, which calls for legislation to reduce tobacco smoke pollution.