Molecular identification of Sarcocystis wobeseri-like parasites in a new intermediate host species, the white-tailed sea eagle (Haliaeetus albicilla).

A reintroduced white-tailed sea eagle (Haliaeetus albicilla) in moderate body condition was found dead and submitted for post-mortem examination. There were no signs of disease on gross pathological examination. Histopathological examination however revealed the presence of encysted protozoan parasites in pectoral and cardiac muscle sections. Polymerase chain reaction amplification of extracted genomic DNA and sequencing of four regions: the 18S rDNA, 28S rDNA, internal transcribed spacer (ITS) 1, and RNA polymerase B (rpoB) loci, confirmed the presence of a Sarcocystis species in pectoral and cardiac muscle which appeared phylogenetically similar to Sarcocystis wobeseri. This is the first report of S. wobeseri-like infection in a white-tailed sea eagle revealing a new intermediate host species for this parasite.

Endothelial activation is associated with intestinal epithelial injury, systemic inflammation and treatment regimen in children living with vertically acquired HIV-1 infection.

BACKGROUND: Premature development of cardiovascular disease in children living with HIV-1 (CLWH) may be associated with compromised gut barrier function, microbial translocation, immune activation, systemic inflammation and endothelial activation. Biomarkers of these pathways may provide insights into pathogenesis of atherosclerotic disease in CLWH. METHODS: This was a cross-sectional study of CLWH enrolled in the multicentre Early Pediatric Initiation-Canadian Child Cure Cohort (EPIC4 ) who were on antiretroviral therapy (ART) with undetectable viral load. Plasma biomarkers of intestinal epithelial injury [intestinal fatty acid binding protein-1 (IFABP)], systemic inflammation [tumour necrosis factor (TNF) and interleukin-6 (IL-6)] and endothelial activation [angiopoietin-2 (Ang2), soluble vascular endothelial growth factor-1 (sVEGFR1) and soluble endoglin (sEng)] were quantified by enzyme-linked immunosorbent assay. Correlation and factor analysis of biomarkers were used to examine associations between innate immune pathways. RESULTS: Among 90 CLWH, 16% of Ang2, 15% of sVEGFR1 and 23% of sEng levels were elevated relative to healthy historic controls. Pairwise rank correlations between the three markers of endothelial activation were statistically significant (ρ = 0.69, ρ = 0.61 and ρ = 0.65, P < 0.001 for all correlations). An endothelial activation index, derived by factor analysis of the three endothelial biomarkers, was correlated with TNF (ρ = 0.47, P < 0.001), IL-6 (ρ = 0.60, P < 0.001) and intestinal fatty acid binding protein-1 (ρ = 0.67, P < 0.001). Current or past treatment with ritonavir-boosted lopinavir (LPV/r) was associated with endothelial activation (odds ratio = 5.0, 95% CI: 1.7-17, P = 0.0020). CONCLUSIONS: Endothelial activation is prevalent in CLWH despite viral suppression with combination ART and is associated with intestinal epithelial injury, systemic inflammation and treatment with LPV/r.

Mucosal-associated invariant T (MAIT) cells are activated in the gastrointestinal tissue of patients with combination ipilimumab and nivolumab therapy-related colitis in a pathology distinct from ulcerative colitis.

The aim of this study was to investigate the pathogenesis of combination ipilimumab and nivolumab-associated colitis (IN-COL) by measuring gut-derived and peripheral blood mononuclear cell (GMNC; PBMC) profiles. We studied GMNC and PBMC from patients with IN-COL, IN-treated with no adverse-events (IN-NAE), ulcerative colitis (UC) and healthy volunteers using flow cytometry. In the gastrointestinal-derived cells we found high levels of activated CD8+ T cells and mucosal-associated invariant T (MAIT) cells in IN-COL, changes that were not evident in IN-NAE or UC. UC, but not IN-C, was associated with a high proportion of regulatory T cells (Treg ). We sought to determine if local tissue responses could be measured in peripheral blood. Peripherally, checkpoint inhibition instigated a rise in activated memory CD4+ and CD8+ T cells, regardless of colitis. Low circulating MAIT cells at baseline was associated with IN-COL patients compared with IN-NAE in one of two cohorts. UC, but not IN-COL, was associated with high levels of circulating plasmablasts. In summary, the alterations in T cell subsets measured in IN-COL-affected tissue, characterized by high levels of activated CD8+ T cells and MAIT cells and a low proportion of Treg , reflected a pathology distinct from UC. These tissue changes differed from the periphery, where T cell activation was a widespread on-treatment effect, and circulating MAIT cell count was low but not reliably predictive of colitis.

Socio-economic differences in cardiovascular disease risk factor prevalence in people with type 2 diabetes in Scotland: a cross-sectional study.

AIM: To describe the association between socio-economic status and prevalence of key cardiovascular risk factors in people with type 2 diabetes in Scotland. METHODS: A cross-sectional study of 264 011 people with type 2 diabetes in Scotland in 2016 identified from the population-based diabetes register. Socio-economic status was defined using quintiles of the area-based Scottish Index of Multiple Deprivation (SIMD) with quintile (Q)1 and Q5 used to identify the most- and least-deprived fifths of the population, respectively. Logistic regression models adjusted for age, sex, health board, history of cardiovascular disease and duration of diabetes were used to estimate odds ratios (ORs) for Q1 compared with Q5 for each risk factor. RESULTS: The mean (sd) age of the study population was 66.7 (12.8) years, 56% were men, 24% were in Q1 and 15% were in Q5. Crude prevalence in Q1/Q5 was 24%/8.8% for smoking, 62%/49% for BMI ≥ 30 kg/m2 , 44%/40% for HbA1c ≥ 58 mmol/mol (7.5%), 31%/31% for systolic blood pressure (SBP) ≥ 140 mmHg, and 24%/25% for total cholesterol ≥ 5 mmol/l, respectively. ORs [95% confidence intervals (CI)] were 3.08 (2.95-3.21) for current smoking, 1.48 (1.44-1.52) for BMI ≥ 30 kg/m2 , 1.11 (1.08-1.15) for HbA1c ≥ 58 mmol/mol (7.5%), 1.03 (1.00-1.06) for SBP ≥ 140 mmHg and 0.87 (0.84-0.90) for total cholesterol ≥ 5 mmol/l. CONCLUSIONS: Socio-economic deprivation is associated with higher prevalence of smoking, BMI ≥ 30 kg/m2 and HbA1c ≥ 58 mmol/mol (7.5%), and lower prevalence of total cholesterol ≥ 5 mmol/l among people with type 2 diabetes in Scotland. Effective approaches to reducing inequalities are required as well as reducing risk factor prevalence across the whole population.

Breastfeeding by Women Living With Human Immunodeficiency Virus in a Resource-Rich Setting: A Case Series of Maternal and Infant Management and Outcomes.

The reduction in human immunodeficiency virus (HIV) transmission through breastmilk with maternal combination antiretroviral therapy (cART) has led many pregnant women living with HIV and healthcare providers to question exclusive formula feeding in resource-rich settings. Here, we describe cART prophylaxis in 3 breastfed infants whose mothers had sustained virologic suppression; all 3 of these infants remained uninfected.

The severity and associated comorbidities of retinopathy of prematurity among micro-premature infants with birth weights less than 750 grams.

OBJECTIVES: To evaluate the characteristics and comorbidities associated with ROP in micro-premature infants and their results. METHODS: This is a retrospective chart review involving multiple intensive care units in Central Texas from 2011 to 2016. Infants were included if birth weight (BW) was≤750 g with confirmed ROP by the International Classification of Retinopathy of Prematurity (ICROP). Neonates were examined and treated with laser ablation or intravitreal ranibizumab (IVR) with subsequent laser treatment, guided by fluorescein angiography, if met treatment criteria defined as type 1 ROP by the Early Treatment of Retinopathy of Prematurity standards. Time to regression was defined clinically. Results were analyzed using chi-squared test. RESULTS: 100 neonates were included in the study. Mean BW was 599 grams and mean gestational age was 24.2 weeks. Forty neonates were classified as type 1 ROP and therefore required intervention; of them 21 received laser alone and 19 required IVR with subsequent laser. Only 2 patients received more than one IVR injection. None of the patients progressed to stage 4 or 5 ROP. CONCLUSIONS: Despite such low birth weights, none of these neonates progressed to stage 4 or 5 ROP likely because of prompt examination and treatment with laser or with IVR and subsequent laser. IVR might serve as a bridge to laser in type 1 ROP allowing some retinal vessel development prior to definitive laser treatment.

Spontaneous Ruptured Pyomyoma in a Nulligravid Female: A Case Report and Review of the Literature.

INTRODUCTION: Pyomyoma, or suppurative leiomyoma, is a rare complication of uterine fibroids. It occurs most commonly in the setting of pregnancy, the immediate postpartum period, or postmenopausal status. It may also arise after recent uterine instrumentation, after uterine artery embolization, or in immunocompromised patients. The most likely cause of pyomyoma is vascular compromise followed by bacterial seeding from direct, hematogenous, or lymphatic spread. Diagnosis is difficult, as the condition is rare, presents with vague symptoms, and is difficult to identify on imaging. Definitive diagnosis is only possible with surgery. Pathology shows a degenerating fibroid with hemorrhage, necrosis, cystic degeneration, and/or inflammatory change. Cultures of the pus contained within often show polymicrobial infection. CASE PRESENTATION: Our patient is a 24-year-old nulligravid female who presented with a surgical abdomen, fever, hypotension, and leukocytosis. She had no significant prior medical or surgical history, no history of uterine instrumentation, and no history of pelvic infection; she was not currently sexually active at the time of presentation. She was taken to the operating room, where she underwent diagnostic laparoscopy. This showed a ruptured pyomyoma originating in the left broad ligament. She then underwent laparoscopic myomectomy. She was transferred to the ICU intubated; she slowly recovered on IV antibiotics and was discharged home on postoperative day 10. DISCUSSION: Pyomyoma is a rare condition and is even rarer in premenopausal patients without recent history of pregnancy or uterine instrumentation. This demonstrates an unusual case of spontaneous pyomyoma in the absence of risk factors, other than a history of known fibroids. Pyomyoma should be considered as a diagnosis in patients with sepsis, history of fibroids, and no other identifiable source of infection.

Incident ischaemic stroke and Type 2 diabetes: trends in incidence and case fatality in Scotland 2004-2013.

AIM: To describe trends in first ischaemic stroke incidence and case fatality in adults with and without a diagnosis of Type 2 diabetes prior to their ischaemic stroke event in Scotland between 2004 and 2013. METHODS: Using population-wide hospital admission, death and diabetes datasets, we conducted a retrospective cohort study. Negative binomial and logistic regression models were used to calculate year-specific incidence and case-fatality rates for people with Type 2 diabetes and for people without diabetes. RESULTS: During 41.0 million person-years of follow-up there were 69 757 ischaemic stroke events. Type 2 diabetes prevalence among patients who experienced ischaemic stroke increased from 13.5% to 20.3% between 2004 and 2013. Stroke incidence rates declined by 2.7% (95% CI 2.4, 3.0) annually for people with and without diabetes [diabetes/year interaction: rate ratio 0.99 (95% CI 0.98, 1.01)]. Type 2 diabetes was associated with an increased risk of ischaemic stroke in men [rate ratio 1.23 (95% CI 1.17, 1.30)] and women [rate ratio 1.41 (95% CI 1.35, 1.48)]. Case-fatality rates were 14.2% and 12.7% in people with Type 2 diabetes and without diabetes, respectively. Case fatality declined by 3.5% (95% CI 2.7, 4.5) annually [diabetes/year interaction: odds ratio 1.01 (95% CI 0.98, 1.02)]. CONCLUSIONS: Ischaemic stroke incidence declined no faster in people with a diagnosis of Type 2 diabetes than in people without diabetes. Increasing prevalence of Type 2 diabetes among stroke patients may mean that declines in case fatality over time will be less marked in the future.

The antiviral role of zinc and metallothioneins in hepatitis C infection.

Metallothioneins (MTs) are small, cysteine-rich proteins characterized by a high affinity for monovalent and divalent cations, such as copper and zinc. Of the four known MT isoforms, only, members of the MT 1 and 2 subfamilies are widely expressed, acting as metal chaperones whose primary role is to mediate intracellular zinc homoeostasis. Metallothioneins are potently induced by heavy metals and other sources of oxidative stress where they facilitate metal binding and detoxification as well as free radical scavenging. Metallothionein expression is well documented in the context of viral infection; however, it remains uncertain whether MTs possess specific antiviral roles or whether induction is merely a consequence of cellular stress. To better understand the role of MTs following hepatitis C virus (HCV) infection, we examined MT expression and localization in vitro and in vivo and used a siRNA knockdown approach to ascertain their antiviral efficacy. We confirmed HCV-driven MT induction in vitro and demonstrated MT accumulation in the nucleus of HCV-infected hepatocytes by immunofluorescence. Using a pan-MT siRNA to knock down all members of the MT1 and MT2 subfamilies, we demonstrate that they are mildly antiviral against the JFH1 strain of HCV in vitro (~1.4 fold increase in viral RNA, P < .05). Furthermore, the antiviral effect of zinc treatment against HCV in vitro was mediated through MT induction (P < .05). Our data suggest a potential benefit of using zinc as a low-cost adjunct to current HCV antiviral therapies and suggest that zinc may facilitate the antiviral role of MTs against other viruses.

The effect of using mobile applications for improving oral hygiene in patients with orthodontic fixed appliances: a randomised controlled trial.

OBJECTIVE: To investigate the effect of using mobile applications active reminders to improve oral hygiene in comparison to verbal oral hygiene instructions. DESIGN: Two-arm parallel randomised controlled trial. SETTING: orthodontic clinics at two branches of a university hospitals of the college of dentistry of Riyadh Colleges of Dentistry and Pharmacy, Riyadh, Saudi Arabia. PARTICIPANTS: Forty-four 12-year-old and older subjects. METHOD: Subjects undergoing orthodontic treatment with fixed appliances were randomly assigned to one of two groups using simple randomisation. Group I: subjects received a mobile application that sends active reminders of oral hygiene three times a day (n = 22). Group II: subjects received verbal oral hygiene instructions verbally during their routine orthodontic visits (n = 22). Two primary outcomes were assessed using plaque index (PI) and gingival index (GI) for Ramfjord teeth to evaluate the level of oral hygiene at baseline and after 4 weeks. RESULTS: Mean differences for PI and GI for group I were reduced from T1 to T2 (P < 0.05, P < 0.05) but did not significantly change for group II (P > 0.05, P > 0.05). Both PI and GI significantly reduced for group I compared to group II between T1 and T2 (P < 0.05, P < 0.05). CONCLUSIONS: PI and GI all significantly decreased after 4 weeks of using active reminders of oral hygiene instructions on mobile application compared to verbal oral hygiene instructions. The study was registered at clinicaltrials.gov with number: NCT03109769.

Assessing knowledge and attitudes about end of life: Evaluation of three instruments designed for adults with intellectual disability.

BACKGROUND: This paper examines the development and psychometric characteristics of three instruments about end of life, designed for use with adults with intellectual disability (ID). Respectively, the instruments assess understanding of the concept of death, end-of-life planning, and fear of death. METHODS: Part 1: instruments were developed or adapted, and pilot tested with 11 adults with ID and 2 disability staff. Part 2: 39 adults with ID and 40 disability staff were assessed on all three instruments. RESULTS: We evaluated comprehensibility, internal consistency, inter-rater reliability, subscale: total score correlations, missing data, and withdrawal. Psychometric findings were mostly good. Overall, 23% of participants with ID withdrew at some point. This outcome may have been as much due to assessment fatigue as to sensitive content. There were no adverse events. CONCLUSIONS: People with ID can reliably complete assessments about end-of-life. Generally, each instrument was found to be comprehensible, reliable and valid.

Associations between neighbourhood environmental factors and the uptake and effectiveness of a brief intervention to increase physical activity: findings from deprived urban communities in an English city.

Background: Evidence suggests behavioural interventions may exacerbate health inequalities, potentially due to differences in uptake or effectiveness. We used a physical activity intervention targeting deprived communities to identify neighbourhood-level factors that might explain differences in programme impact. Methods: Individuals aged 40-65 were sent a postal invitation offering a brief intervention to increase physical activity. We used postcodes linkage to determine whether neighbourhood indicators of deprivation, housing, crime and proximity to green spaces and leisure facilities predicted uptake of the initial invitation or an increase in physical activity level in those receiving the brief intervention. Results: A total of 4134 (6.8%) individuals responded to the initial invitation and of those receiving the intervention and contactable after 3 months, 486 (51.6%) reported an increase in physical activity. Area deprivation scores linked to postcodes predicted intervention uptake, but not intervention effectiveness. Neighbourhood indicators did not predict either uptake or intervention effectiveness. Conclusions: The main barrier to using brief intervention invitations to increase physical activity in deprived, middle-aged populations was the low uptake of an intervention requiring significant time and motivation from participants. Once individuals have taken up the intervention offer, neighbourhood characteristics did not appear to be significant barriers to successful lifestyle change.

IFNL3/4 genotype is associated with altered immune cell populations in peripheral blood in chronic hepatitis C infection.

Single-nucleotide polymorphisms near the interferon lambda 3 (IFNL3) gene predict outcomes to infection and anti-viral treatment in hepatitis C virus (HCV) infection. To identify IFNL3 genotype effects on peripheral blood, we collected phenotype data on 400 patients with genotype 1 chronic hepatitis C (CHC). The IFNL3 responder genotype predicted significantly lower white blood cells (WBCs), as well as lower absolute numbers of monocytes, neutrophils and lymphocytes for both rs8099917 and rs12979860. We sought to define the WBC subsets driving this association using flow cytometry of 67 untreated CHC individuals. Genotype-associated differences were seen in the ratio of CD4CD45RO+ to CD4CD45RO-; CD8CD45RO+ to CD8CD45RO-, NK CD56 dim to bright and monocyte numbers and percentages. Whole blood expression levels of IFNL3, IFNLR1 (interferon lambda receptor 1), IFNLR1-mem (a membrane-associated receptor), IFNLR1-sol (a truncated soluble receptor), MxA and T- and NK (natural killer) cell transcription factors TBX21, GATA3, RORC, FOXP3 and EOMES in two subjects were also determined. CHC patients demonstrated endogenous IFN activation with higher levels of MxA, IFNLR1, IFNLR1-mem and IFNLR1-sol, and IFNL3 genotype-associated differences in transcription factors. Taken together, these data provide evidence of an IFNL3 genotype association with differences in monocyte, T- and NK cell levels in the peripheral blood of patients with CHC. This could underpin genotype associations with spontaneous and treatment-induced HCV clearance and hepatic necroinflammation.

Variations in weight management services in Scotland: a national survey of weight management provision.

BACKGROUND: Despite National guidance recommending their use, there is uncertainty regarding the best way to deliver weight management services across the UK and worldwide. METHODS: To ascertain access, provision and interventions used in lifestyle Tier 2 and specialist Tier 3 weight management services in Scotland, a survey was distributed to all mainland health boards covering pathways for referral, eligibility criteria, intervention format and definitions of attendance completion and adherence. RESULTS: Nine Health boards provided information on their weight management services. The provision of services was low. Only four health boards offered services for those with a BMI 25-30 kg/m2. Lifestyle Tier 2 services were mainly weekly or fortnightly group sessions for 8-12 weeks delivered by dietitians or community workers. Specialist Tier 3 services were largely similar to lifestyle Tier 2 services. The provision of specialist interventions including pharmacotherapy, cognitive behavioural therapy sessions and low-calorie prescribed diets was low. CONCLUSIONS: This national survey has illustrated large disparities in the provision of weight management across Scotland, a likely consequence of uncertainty regarding best practice. There is a clear requirement for the evaluation of existing services to identify those that lead to the largest improvements in health outcomes and are cost-effective.

Hepatic metallothionein expression in chronic hepatitis C virus infection is IFNL3 genotype-dependent.

The IFNL3 genotype predicts the clearance of hepatitis C virus (HCV), spontaneously and with interferon (IFN)-based therapy. The responder genotype is associated with lower expression of interferon stimulated genes (ISGs) in liver biopsies from chronic hepatitis C patients. However, ISGs represent many interacting molecular pathways, and we hypothesised that the IFNL3 genotype may produce a characteristic pattern of ISG expression explaining the effect of genotype on viral clearance. For the first time, we identified an association between a cluster of ISGs, the metallothioneins (MTs) and IFNL3 genotype. Importantly, MTs were significantly upregulated (in contrast to most other ISGs) in HCV-infected liver biopsies of rs8099917 responders. An association between lower fibrosis scores and higher MT levels was demonstrated underlying clinical relevance of this association. As expected, overall ISGs were significantly downregulated in biopsies from subjects with the IFNL3 rs8099917 responder genotype (P=2.38 × 10(-7)). Peripheral blood analysis revealed paradoxical and not previously described findings with upregulation of ISGs seen in the responder genotype (P=1.00 × 10(-4)). The higher MT expression in responders may contribute to their improved viral clearance and MT-inducing agents may be useful adjuncts to therapy for HCV. Upregulation of immune cell ISGs in responders may also contribute to the IFNL3 genotype effect.

Pharmacological validation of a refined burrowing paradigm for prediction of analgesic efficacy in a rat model of sub-chronic knee joint inflammation.

BACKGROUND: Burrowing is an evolutionarily conserved behaviour in rodents. This study validates a refined burrowing paradigm (requiring a reduced number of animals) in a rat model of sub-chronic knee joint inflammation and evaluates its sensitivity and specificity for analgesic drugs. METHODS: Knee joint inflammation in rats was induced by intra-articular injection with complete Freund's adjuvant (CFA). Burrowing performance was assessed at baseline without study drugs, and in CFA-naive and CFA-injected animals following administration of the analgesic drugs naproxen, pregabalin and morphine, each at three doses, or corresponding vehicle (nine rats per dose group). The specificity of the model was evaluated by also testing the anxiogenic drug yohimbine, the stimulant drug dexamphetamine and the anxiolytic drug chlordiazepoxide in CFA-naive and CFA-injected animals. Percentage maximum possible effect (%MPE) was determined by relating the difference between post-CFA and baseline burrowing performance in each drug dose group to that in the vehicle group in each experiment. RESULTS: Burrowing performance in the vehicle groups was decreased by 39.0-59.8% in CFA-injected animals compared with CFA-naive animals. CFA-induced reductions in burrowing performance were reversed by each of the three analgesic drugs tested. The highest %MPE was 75.2% with naproxen 50 mg/kg, 80.9% with pregabalin 10 mg/kg and 77.0% with morphine 1 mg/kg (all p < 0.05 vs. control). CFA-induced reductions in burrowing performance were not reversed by yohimbine, dexamphetamine or chlordiazepoxide. CONCLUSIONS: This study provides pharmacological validation of a refined burrowing paradigm for analgesic efficacy that exhibits good predictive validity, with high sensitivity and specificity.

Burrowing as a non-reflex behavioural readout for analgesic action in a rat model of sub-chronic knee joint inflammation.

BACKGROUND: Innate responses against spontaneous pain are proposed to improve the predictive validity of preclinical analgesia models. Therefore, development and validation of novel readouts is necessary. To investigate whether innate rodent burrowing is a useful alternative behavioural readout for assessment of analgesic efficacy, a complete Freund's adjuvant (CFA)-induced model of sub-chronic inflammation was used to compare the effects of naproxen, ibuprofen and pregabalin in weight-bearing (WB), open-field (OF) and burrowing assays. METHODS: Male Sprague Dawley rats were injected with 150 µL of CFA (2 mg/mL) into the knee (hind leg) 3 days before testing. Naproxen, ibuprofen and pregabalin were administered at different doses 30, 90 and 60 min, respectively, before testing. WB was determined using a rat incapacitance tester; horizontal distance moved and vertical rearings were recorded in an OF; and burrowing was measured by the weight of gravel remaining in a hollow tube after 60 min. RESULTS: CFA-induced arthritis reduced WB, OF activity and burrowing. Naproxen, pregabalin and ibuprofen treatment normalized WB; however, horizontal OF activity was not improved by any treatment; rearing behaviour was moderately reinstated by ibuprofen (100 mg/kg). In burrowing, naproxen (100 mg/kg), ibuprofen (31.6 and 100 mg/kg) and pregabalin (10 mg/kg) reversed CFA-induced deficits. CONCLUSIONS: Burrowing performance is an alternative non-reflex readout relying on innate rodent behaviour that is affected by nociceptive behaviour and can be pharmacologically manipulated. The burrowing assay appears to be more sensitive than OF assays and is as sensitive as WB assays at distinguishing between analgesic doses and doses that impair locomotion.

A novel gene therapy strategy using secreted multifunctional anti-HIV proteins to confer protection to gene-modified and unmodified target cells.

Current human immunodeficiency virus type I (HIV) gene therapy strategies focus on rendering HIV target cells non-permissive to viral replication. However, gene-modified cells fail to accumulate in patients and the virus continues to replicate in the unmodified target cell population. We have designed lentiviral vectors encoding secreted anti-HIV proteins to protect both gene-modified and unmodified cells from infection. Soluble CD4 (sCD4), a secreted single chain variable fragment (sscFv(17b)) and a secreted fusion inhibitor (sFI(T45)) were used to target receptor binding, co-receptor binding and membrane fusion, respectively. Additionally, we designed bi- and tri-functional fusion proteins to exploit the multistep nature of HIV entry. Of the seven antiviral proteins tested, sCD4, sCD4-scFv(17b), sCD4-FI(T45) and sCD4-scFv(17b)-FI(T45) efficiently inhibited HIV entry. The neutralization potency of the bi-functional fusion proteins sCD4-scFv(17b) and sCD4-FI(T45) was superior to that of sCD4 and the Food and Drug Administration-approved fusion inhibitor T-20. In co-culture experiments, sCD4, sCD4-scFv(17b) and sCD4-FI(T45) secreted from gene-modified producer cells conferred substantial protection to unmodified peripheral blood mononuclear cells. In conclusion, continuous delivery of secreted anti-HIV proteins via gene therapy may be a promising strategy to overcome the limitations of the current treatment.

A study on the use of Gumbel approximation with the Bernoulli spatial scan statistic.

The Bernoulli version of the spatial scan statistic is a well established method of detecting localised spatial clusters in binary labelled point data, a typical application being the epidemiological case-control study. A recent study suggests the inferential accuracy of several versions of the spatial scan statistic (principally the Poisson version) can be improved, at little computational cost, by using the Gumbel distribution, a method now available in SaTScan(TM) (www.satscan.org). We study in detail the effect of this technique when applied to the Bernoulli version and demonstrate that it is highly effective, albeit with some increase in false alarm rates at certain significance thresholds. We explain how this increase is due to the discrete nature of the Bernoulli spatial scan statistic and demonstrate that it can affect even small p-values. Despite this, we argue that the Gumbel method is actually preferable for very small p-values. Furthermore, we extend previous research by running benchmark trials on 12 000 synthetic datasets, thus demonstrating that the overall detection capability of the Bernoulli version (i.e. ratio of power to false alarm rate) is not noticeably affected by the use of the Gumbel method. We also provide an example application of the Gumbel method using data on hospital admissions for chronic obstructive pulmonary disease.

Early caffeine therapy and clinical outcomes in extremely preterm infants.

OBJECTIVE: To determine if early caffeine (EC) therapy is associated with decreased bronchopulmonary dysplasia (BPD) or death, decreased treatment of patent ductus arteriosus (PDA), or shortened duration of ventilation. STUDY DESIGN: In a retrospective cohort of 140 neonates ≤1250 g at birth, infants receiving EC (initiation <3 days of life) were compared with those receiving late caffeine (LC, initiation ≥3 days of life) using logistic regression. RESULT: Of infants receiving EC, 25% (21/83) died or developed BPD compared with 53% (30/57) of infants receiving LC (adjusted odds ratio (aOR) 0.26, 95% confidence interval (CI) 0.09 to 0.70; P<0.01). PDA required treatment in 10% of EC infants versus 36% of LC infants (aOR 0.28, 95%CI 0.10 to 0.73; P=0.01). Duration of mechanical ventilation was shorter in infants receiving EC (EC, 6 days; LC, 22 days; P<0.01). CONCLUSION: Infants receiving EC therapy had improved neonatal outcomes. Further studies are needed to determine if caffeine prophylaxis should be recommended for preterm infants.