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.

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.

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.

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.

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.

Hospitalisations and costs relating to ambulatory care sensitive conditions in Ireland.

BACKGROUND: Ambulatory care sensitive conditions (ACSCs) are conditions for which the provision of timely and effective outpatient care can reduce the risks of hospitalisation by preventing, controlling or managing a chronic disease or condition. AIMS: The aims of this study were to report on ACSCs in Ireland, and to provide a baseline for future reference. METHODS: Using HIPE, via Health Atlas Ireland, inpatient discharges classified as ACSCs using definitions from the Victorian ACSC study were extracted for the years 2005-2008. Direct methods of standardisation allowed comparison of rates using the EU standard population as a comparison for national data, and national population as comparison for county data. Costs were estimated using diagnosis-related groups. RESULTS: The directly age-standardised discharge rate for ACSC-related discharges increased slightly, but non-significantly, from 15.40 per 1,000 population in 2005 to 15.75 per 1,000 population in 2008. The number of discharges increased (9.5%) from 63,619 in 2005 to 69,664 in 2008, with the estimated associated hospital costs increasing (31.5%) from 267.8 million in 2005 to 352.2 million in 2008. Across the country, there was considerable variation in the discharge rates for the Top-10 ACSCs for the years 2005-2008. Significantly lower rates of hospitalisation were observed in more urban areas including Cork, Dublin and Galway. The most common ACSC in 2008 was diabetes with complications (29.8%). CONCLUSIONS: The variation in rates observed indicates the scope of reducing hospitalisations and associated costs for ACSCs, across both adult's and children's services and particularly in relation to diabetes complications.

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.

MicroRNA expression profiling identifies activated B cell status in chronic lymphocytic leukemia cells.

Chronic lymphocytic leukemia (CLL) is thought to be a disease of resting lymphocytes. However, recent data suggest that CLL cells may more closely resemble activated B cells. Using microRNA (miRNA) expression profiling of highly-enriched CLL cells from 38 patients and 9 untransformed B cells from normal donors before acute CpG activation and 5 matched B cells after acute CpG activation, we demonstrate an activated B cell status for CLL. Gene set enrichment analysis (GSEA) identified statistically-significant similarities in miRNA expression between activated B cells and CLL cells including upregulation of miR-34a, miR-155, and miR-342-3p and downregulation of miR-103, miR-181a and miR-181b. Additionally, decreased levels of two CLL signature miRNAs miR-29c and miR-223 are associated with ZAP70(+) and IgV(H) unmutated status and with shorter time to first therapy. These data indicate an activated B cell status for CLL cells and suggest that the direction of change of individual miRNAs may predict clinical course in CLL.

Macrovertebrate paleontology and the Pliocene habitat of Ardipithecus ramidus.

A diverse assemblage of large mammals is spatially and stratigraphically associated with Ardipithecus ramidus at Aramis. The most common species are tragelaphine antelope and colobine monkeys. Analyses of their postcranial remains situate them in a closed habitat. Assessment of dental mesowear, microwear, and stable isotopes from these and a wider range of abundant associated larger mammals indicates that the local habitat at Aramis was predominantly woodland. The Ar. ramidus enamel isotope values indicate a minimal C4 vegetation component in its diet (plants using the C4 photosynthetic pathway), which is consistent with predominantly forest/woodland feeding. Although the Early Pliocene Afar included a range of environments, and the local environment at Aramis and its vicinity ranged from forests to wooded grasslands, the integration of available physical and biological evidence establishes Ar. ramidus as a denizen of the closed habitats along this continuum.

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.

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.

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.

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.

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.

A small RNA makes a Bic difference.

The first highly specific knockouts of a microRNA, miR155, in mice result in multiple defects in adaptive immunity, and also show the feasibility of investigating at least some microRNAs by gene knockout.

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.

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.

Blood alcohol levels in persons who died from accidents and suicide.

Although it is known that alcohol is associated with a high proportion of fatal accidents and suicides, little information is available in Ireland on blood alcohol concentrations (BACs) of those who died. This study was undertaken to identity the (BACs) in persons who died as a result of suicide or injury. The study was a retrospective review of coroners' records to identify BACs in three counties in Ireland. All cases where the person died as a result of injury or suicide in 2001 and 2002 were included. There were 129 deaths eligible for inclusion. Of these, 98 (76%) were male, 55 (42.6%) were road traffic accidents (RTAs), 31 (24.0%) suicides, 12 (9.3%) substance misuse, 11 (8.5%) house fires and 20 (15.5%) others. Of the 55 who died as a result of RTAs, 22 (40%) had positive BACs ranging from 16mg/100 ml to 325 mg/100 ml. Of the 31 who died as a result of suicide, 28 (90.3%) were male. BACs were available for 29 (93.5%). Of these, 16 (55.5%) had alcohol detected. Persons aged less than 30 years were more likely to have alcohol in their blood (p < 0.002). The mean BAC for persons aged less than 30 was 191.5 mg/100 ml compared to 84.0 mg/100 ml for those aged 30 and over. The mean BAC for adults who died in house fires was 225.2 mg/100 ml. The high BACs in those who died as a result of suicide or injury reflect the high level of alcohol consumption and binge drinking in Ireland.