End-to-end sensor and neural network design using differential ray tracing.

In this paper we propose a new method to jointly design a sensor and its neural-network based processing. Using a differential ray tracing (DRT) model, we simulate the sensor point-spread function (PSF) and its partial derivative with respect to any of the sensor lens parameters. The proposed ray tracing model makes no thin lens nor paraxial approximation, and is valid for any field of view and point source position. Using the gradient backpropagation framework for neural network optimization, any of the lens parameter can then be jointly optimized along with the neural network parameters. We validate our method for image restoration applications using three proves of concept of focus setting optimization of a given sensor. We provide here interpretations of the joint optical and processing optimization results obtained with the proposed method in these simple cases. Our method paves the way to end-to-end design of a neural network and lens using the complete set of optical parameters within the full sensor field of view.

Impairment of Nrf2- and Nitrergic-Mediated Gastrointestinal Motility in an MPTP Mouse Model of Parkinson's Disease.

BACKGROUND: Gastrointestinal (GI) motility dysfunction is the most common non-motor symptom of Parkinson's disease (PD). Studies have indicated that GI motility functions are impaired before the onset of PD. AIMS: To investigate the underlying mechanism of PD-induced GI dysmotility in MPTP (1-methyl 4-phenyl 1,2,3,6-tetrahydropyridine)-induced animal model. METHODS: C57BL/6 mice were administered with or without a selective dopamine neurotoxin, MPTP, to induce parkinsonian symptoms. In addition to in vivo studies, in vitro experiments were also conducted in colon specimens using l-methyl-4-phenylpyridinium (MPP+), a metabolic product of MPTP. Gastric emptying, colon motility, nitrergic relaxation, and western blot experiments were performed as reported. RESULTS: MPTP-induced PD mice showed decreased expression of nuclear factor erythroid 2-related factor (Nrf2) and its target phase II genes in gastric and colon neuromuscular tissues. Decreased levels of tetrahydrobiopterin (BH4, a critical cofactor for nNOS dimerization) associated with uncoupling of nNOS in gastric and colon tissues exposed to MPTP. Impaired enteric nitrergic system led to delayed gastric emptying and slower colonic motility compared to the control mice. In vitro results in colon specimens confirm that activation of Nrf2 restored MPP+-induced suppression of alpha-synuclein, tyrosine hydroxylase (TH), Nrf2, and heme oxygenase-1. In vitro exposure to L-NAME [N(w)-nitro-L-arginine methyl ester], a NOS synthase inhibitor, reduced protein expression of TH in colon tissue homogenates. CONCLUSIONS: Loss of Nrf2/BH4/nNOS expression in PD impairs antioxidant gene expression, which deregulates NO synthesis, thereby contributing to the development of GI dysmotility and constipation. Nitric oxide appears to be important to maintain dopamine synthesis in the colon.

The effect of different irrigation protocols on elastic modulus of dentine and biomechanics of single-rooted premolar tooth: A nano-indentation and finite element analysis study.

AIM: This study evaluated the effect of different irrigation protocols on elastic modulus and biomechanics of single-rooted premolar tooth using with nano-indentation and finite element analysis (FEA). MATERIALS AND METHODS: Root canals of single-rooted human teeth were prepared, divided into eight groups, and irrigated with (1) 2.5% NaOCl + 17% EDTA; (2) 2.5% NaOCl + 17% EDTA + 2.5% NaOCl; (3) 2.5% NaOCl + SmearClear; (4) 2.5% NaOCl + 2% chlorhexidine; (5) 1.3% NaOCl + MTAD; (6) 5.25% NaOCl; (7) 17% EDTA; and (8) saline. The roots were vertically sectioned, and elastic modulus of the root dentine was measured using nano-indenter device at coronal, middle, and apical third. Data were recorded as megapascal and statistically analyzed (one-way analysis of variance, Tukey tests). Three-dimensional FEA model of a premolar tooth was created, and the inner root dentine was modified to simulate the effect of irrigation protocols on root dentine. The elastic properties of inner root dentine layer in the FEA models were modified for each group according to the data obtained with nano-indentation. A 300-N load was applied at the buccal cusp and central fossa of the models with a 45° angle. The stresses were calculated using von Mises stress criteria. RESULTS: All irrigation protocols affected the elastic modulus of root dentine. Groups 2 and 3 showed similar elastic modulus values (P > 0.05), whereas the lowest values were obtained in group 7 (P < 0.05). No statistically significant differences were found between groups 4, 5, and 8 (P > 0.05). CONCLUSION: Despite the effect of different clinically used irrigation protocols on elastic modulus of the inner dentine, this does not affect the biomechanics of the roots.

Chronic estrogen deficiency causes gastroparesis by altering neuronal nitric oxide synthase function.

BACKGROUND: Gastroparesis affects predominantly females; however, the biological basis for this gender bias is completely unknown. Several lines of evidence suggest that nitrergic dependent stomach motility function is reduced in diabetic gastroparesis and that nNOS is estrogen-regulated. AIMS: The purpose of this study was to investigate whether reduced levels of estradiol-17ß (E2) down-regulates tetrahydrobiopterin (BH4, a cofactor for nNOS dimerization and enzyme activity) biosynthesis and therefore nNOS mediated gastric motility would be impaired in a mouse model of chronic estrogen deficiency, follicle stimulating hormone receptor knock-out female mice (FORKO). METHODS: In-bred 12-week-old female FORKO mice were obtained from our FORKO breeding colony. Gastric emptying was measured in overnight fasting mice. Nitrergic relaxation (AUC/mg tissue) was measured at 2 Hz through electric field stimulation using gastric antrum strips prepared from WT and FORKO mice. Protein expression for nNOSα, BH4 biosynthesis enzymes (GCH-1, DHFR) and estrogen receptors (α, ß) were measured in gastric antrum by western blotting. Levels of BH4 and oxidized BH2, B biopterin levels were determined by HPLC. RESULTS: In FORKO, compared to wild type (WT) stomachs we indentified (1) reduced (%) gastric emptying (64 ± 2.5 vs. 77.6 ± 0.88), (2) greater reduction in nitregic relaxation (-0.13 ± 0.012 vs. -0.28 ± 0.012), (3) increased nNOS dimerization (0.48 ± 0.02 vs. 0.34 ± 0.05), (4) decreased NO release whether measured at 24 h (0.6 ± 0.04 vs. 1.7 ± 0.22, p < 0.05) or at 48 h (3.4 ± 0.26 vs. 5.0 ± 0.15, p < 0.05) of incubation, (5) decreased GCH-1 (1.9 ± 0.06 vs. 2.3 ± 0.04), DHFR (1.8 ± 0.14 vs. 2.4 ± 0.07) and ERα (2.7 ± 0.4 vs. 3.9 ± 0.4) and (6) increased oxidized biopterin levels and decreased ratio of BH4 versus BH2 + B. CONCLUSION: We conclude that chronic estrogen deficiency negatively modifies the function of both BH4 and nNOS thereby contributing to the development of gastroparesis in a FORKO mouse model.

First Report of Sorghum mosaic virus Causing Mosaic in Miscanthus sinensis.

Miscanthus is being evaluated as a bioenergy feedstock because of its potentially significant biomass production, perennial habit, and lack of major diseases and pests. It is also a valuable parent in sugarcane breeding programs as a source of cold tolerance. In May 2010, mosaic symptoms were observed on a clone of Miscanthus sinenesis Anderss. maintained at the USDA, ARS, Sugarcane Research Unit. All plants of the Miscanthus clone in our germplasm collection are from vegetative cuttings of the original infected plant and show mosaic symptoms. Leaves from the ratoon of a single plant tested positive in a reverse transcription-PCR with the Potyvirus Group PCR Test (Agdia, Inc., Elkhart, IN) with two primer sets, Poty-F1/Poty-R1 and Poty-F2/Poty-R2. After sequencing the potyvirus amplicons, a BLAST search in GenBank revealed that these sequences had the highest identities (81 and 69%) with Sorghum mosaic virus (SrMV) at the nucleic acid level and a 72 and 95% similarity at the amino acid level. Extracts from the Miscanthus clone prepared by the indirect extraction buffer (Agdia) containing sodium carbonate also tested positive for potyvirus by indirect ELISA with the 'universal' potyvirus monoclonal antibody, PTY1. To our knowledge, this is the first report of SrMV on Miscanthus. The only other member of the genus Potyvirus reported on Miscanthus is Sugarcane mosaic virus (1,2). Mosaic caused by SrMV could become an economically important disease of Miscanthus if this crop is grown for bioenergy feedstock on large areas. An SrMV-infected Miscanthus crop could pose a threat to established crops of susceptible sugarcane and sorghum since the virus is transmitted in a nonpersistent manner by several aphids, as well as, contributing to geographic shifts of the pathogen. References: (1) B. O. Agindotan et al. J. Virol. Methods 169:119, 2010. (2) D.-L. Xu et al. Arch. Virol. 153:1031, 2008.

An evaluation of a collaborative pharmacist prescribing model compared to the usual medical prescribing model in the emergency department.

BACKGROUND: Workforce reform has placed a significant focus on the role of non-medical prescribers in the healthcare system. Pharmacists are trained in pharmacology and therapeutics, and therefore well placed to act as non-medical prescribers. OBJECTIVES: To assess the safety and accuracy of inpatient medication charts within a pharmacist collaborative prescribing model (intervention), compared to the usual medical model (control) in the emergency department (ED). Another objective compared venous thromboembolism (VTE) risk assessment and prescribing, between intervention and control groups. METHODS: Adult patients in ED referred for hospital admission were randomised into control or intervention by a block randomisation method, until the required sample size was reached. Medication charts were audited retrospectively by an independent auditor, using validated audit forms. RESULTS: Intervention group medication charts contained significantly fewer prescribing errors, omissions and discrepancies compared to the control group, and improved documentation of adverse drug reactions. VTE risk assessment and prescribing had higher guideline concordance in the intervention group compared to the control group. CONCLUSIONS: This collaborative prescribing trial showed excellent results in safety and accuracy of pharmacist prescribing when compared to the usual medical model of prescribing. The admitting medical practitioner and extended scope pharmacist prescriber worked as a collaborative team in emergency, which improved Australian national prescribing safety indicators.

Synthetic phospholipid vesicles containing a purified viral antigen and cell membrane proteins stimulate the development of cytotoxic T lymphocytes.

Synthetic phospholipid vesicles (liposomes) containing the purified glycoprotein (G) of vesicular stomatitis virus (VSV) and solubilized membrane proteins from cells of the appropriate H-2 haplotype elicited H-2-restricted cytotoxic T lymphocytes (CTL) that lysed VSV-infected target cells. The CTL were elicited by intact liposomes, not by released components. Thus, when spleen cells from VSV-primed H-2d X H-2b hybrid mice were stimulated with liposomes having G protein + membrane proteins from cells with one of the parental H-2 haplotypes, the resulting CTL lysed only VSV-infected target cells with that parent's H-2 type. This result argues against the view that T cells in general recognize only processed antigenic fragments on macrophages. Moreover, liposomes were only effective when G protein and cell membrane proteins were included in the same vesicles. This result suggests that for effective interaction with CTL precursors the antigen (G protein) and products of the H-2 complex must be closer to each other than 600--1,000 angstrom, the diameter of the lipid vesicles used in this study.

Comprehensive genetic analysis of the platelet activating factor acetylhydrolase (PLA2G7) gene and cardiovascular disease in case-control and family datasets.

Platelet-activating factor acetylhydrolase (PLA2G7) is a potent pro- and anti-inflammatory molecule that has been implicated in multiple inflammatory disease processes, including cardiovascular disease. The goal of this study was to investigate the genetic effects of PLA2G7 on coronary artery disease (CAD) risk in two large, independent datasets with CAD. Using a haplotype tagging (ht) approach, 19 ht single nucleotide polymorphisms (SNPs) were genotyped in CATHGEN case-control samples (cases = 806 and controls = 267) and in the GENECARD Family Study (n = 1101 families, 2954 individuals). Single SNP analysis using logistic regression revealed nine SNPs with significant association in all CATHGEN subjects (P = 0.0004-0.02). CATHGEN cases were further stratified into subgroups based on age of CAD onset (AOO) and severity of disease; 599 young affecteds (YA, AOO <56) and 207 old affected (OA, AOO >56). Significant genetic effects were observed in both OA and YA (P = 0.0001-0.02). The GENECARD probands demonstrated results similar to those seen in the YA CATHGEN cases (P = 0.002-0.05). Of the 19 SNPs genotyped, 3 SNPs result in nonsynonymous coding changes (I198T, A379V and R92H). Two of the coding SNPs, R92H and A379V, constitute two of the most significantly associated SNPs, even after Bonferroni correction and appear to represent independent associations (r(2) = 0.09). Multiple additional polymorphisms in low linkage disequilibrium with these coding SNPs were also strongly associated. In summary, PLA2G7 represents an important, potentially functional candidate in the pathophysiology of CAD based on replicated associations using two independent datasets and multiple statistical approaches. Further functional studies involving a combination of risk alleles are warranted.

Physical properties of MTA, BioAggregate and Biodentine in simulated conditions: A micro-CT analysis.

The aim of this study was to evaluate the physical properties of calcium-silicate based cements as retrograde filling in different pH and blood conditions using micro-CT. Eighty-four teeth were obturated and after root-end resection, 3 mm-deep root-end cavity was ultrasonically prepared. The samples were divided into four test groups (n=21). Cements were freshly prepared and analyzed with micro-CT before and after exposed. The test groups were exposed for four days to environments containing acid, alkali, or blood. An acidic pH significantly reduced the volume of all materials after four days and differed from the other environments. Biodentine has the highest percentage of loss in volume and density after treatment in an acidic environment. Porosity formation in the acidic medium was observed in Biodentine amongst all materials. The three-dimensional structures of all materials changed after exposed to an acidic pH, while fewer changes were observed in the structures of materials treated with blood and alkali.

Genomic characterization of Ugandan smallholder farmer-preferred cassava varieties.

Understanding the genetic relationships among farmer-preferred cassava (Manihot esculenta Crantz) varieties is indispensable to genetic improvement efforts. In this study, we present a genetic analysis of 547 samples of cassava grown by 192 smallholder farmers, which were sampled at random within four districts in Uganda. We genotyped these samples at 287,952 single nucleotide polymorphisms using genotyping-by-sequencing and co-analyzed them with 349 cassava samples from the national breeding program in Uganda. The samples collected from smallholders consisted of 86 genetically unique varieties, as assessed using a genetic distance-based approach. Of these varieties, most were cultivated in only one district (30 in Kibaale, 19 in Masindi, 14 in Arua, and three in Apac), and only three were cultivated across all districts. The genetic differentiation we observed among farming districts in Uganda (mean fixation index [F ST] = .003) is similar to divergence observed within other countries. Despite the fact that none of the breeding lines were directly observed in farmer fields, genetic divergence between the populations was low (F ST = .020). Interestingly, we detected the presence of introgressions from the wild relative M. glaziovii Müll. Arg. on chromosomes 1 and 4, which implies ancestry with cassava breeding lines. Given the apparently similar pool of alleles in the breeding germplasm, it is likely that breeders have the raw genetic material they require to match the farmer-preferred trait combinations necessary for adoption. Our study highlights the importance of understanding the genetic makeup of cassava currently grown by smallholder farmers and relative to that of plant breeding germplasm.

Wheat blast: histopathology and transcriptome reprogramming in response to adapted and nonadapted Magnaporthe isolates.

* Blast disease (causal agent Magnaporthe oryzae) has presented as a new and serious field disease of wheat in South America. Here, we investigated the responses of wheat to both adapted and nonadapted isolates of the blast fungus Magnaporthe, examining cellular defence and transcriptional changes. * Resistance towards the nonadapted isolate was associated with the formation of appositions, here termed halos, beneath attempted Magnaporthe grisea penetration sites that wheat-adapted, M. oryzae isolates were able to breach. * Transcriptome analysis indicated extensive transcriptional reprogramming following inoculation with both wheat-adapted and nonadapted isolates of Magnaporthe. Functional annotation of many of the differentially expressed transcripts classified into the categories: cell rescue and defence, plant metabolism, cellular transport and regulation of transcription (although a significant number of transcripts remain unclassified). * Defence-related transcripts induced in common by adapted and nonadapted isolates were differentially regulated in response to M. oryzae and M. grisea isolates over time. Differential expression of genes involved in cellular transport indicated the importance of this process in plant defence. Functional characterisation of these transcripts and their role in defence may eventually lead to the identification of broad-spectrum resistance mechanisms in wheat towards Magnaporthe.

Cell surface changes and Rous sarcoma virus gene expression in synchronized cells.

We have investigated whether cell surface changes associated with growth control and malignant transformation are linked to the cell cycle. Chicken embryo cells synchronized by double thymidine block were examined for cell-cycle-dependent alterations in membrane function (measured by transport of 2-deoxyglucose, uridine, thymidine, and mannitol), in cell surface morphology (examined by scanning electron microscopy), and in the ability of tumor virus gene expression to induce a transformation-specific change in membrane function. We reach the following conclusions: (a) The high rate of 2-deoxyglucose transport seen in transformed cells and the low rates of 2-deoxyglucose and uridine transport characteristic of density-inhibited cells do not occur in normal growing cells as they traverse the cell cycle. (b) Although there are cell cycle-dependent changes in surface morphology, they are not reflected in corresponding changes in membrane function. (c) Tumor virus gene expression can alter cell membrane function at any stage in the cell cycle and without progression through the cell cycle.

Endocarditis in mice infected with Coxsackie virus B4.

Endocarditis has not been generally considered to be a complication of viral infection. We show that mural and valvular endocarditis can be produced in mice infected with Coxsackie virus B(4). Because this virus commonly infects man and is highly cardiotropic, it is important to know whether it produces valvular lesions in man similar to those we describe in mice.

GATA2 is associated with familial early-onset coronary artery disease.

The transcription factor GATA2 plays an essential role in the establishment and maintenance of adult hematopoiesis. It is expressed in hematopoietic stem cells, as well as the cells that make up the aortic vasculature, namely aortic endothelial cells and smooth muscle cells. We have shown that GATA2 expression is predictive of location within the thoracic aorta; location is suggested to be a surrogate for disease susceptibility. The GATA2 gene maps beneath the Chromosome 3q linkage peak from our family-based sample set (GENECARD) study of early-onset coronary artery disease. Given these observations, we investigated the relationship of several known and novel polymorphisms within GATA2 to coronary artery disease. We identified five single nucleotide polymorphisms that were significantly associated with early-onset coronary artery disease in GENECARD. These results were validated by identifying significant association of two of these single nucleotide polymorphisms in an independent case-control sample set that was phenotypically similar to the GENECARD families. These observations identify GATA2 as a novel susceptibility gene for coronary artery disease and suggest that the study of this transcription factor and its downstream targets may uncover a regulatory network important for coronary artery disease inheritance.

Lack of evidence of pregnancy-induced alloantibodies in dogs.

BACKGROUND: It is controversial whether or not pregnant bitches become sensitized to red blood cell (RBC) antigens. HYPOTHESIS: Bitches do not develop alloantibodies to RBC antigens during gestation and can be used safely as blood donors. ANIMALS: The study group included 35 healthy female dogs with a prior history of 1 (n = 12), 2 (n = 14), or >or= 3 (n = 9) pregnancies. The control group consisted of 15 healthy female dogs without any history of pregnancy. METHODS: All dogs were blood typed for dog erythrocyte antigens (DEA) 1.1, 1.2, 3, 4, 5, and 7 using ethylenediaminetetraacetic acid blood samples and polyclonal antisera. Antibody screening was performed with serum and canine RBC panels of known blood type. An autocontrol and direct antiglobulin test were performed to rule out the presence of autoantibodies. RESULTS: The only alloantibodies identified were those against DEA 7 and the prevalence of anti-DEA 7 alloantibodies was similar in dogs with known history of pregnancy (11.4%) and in the control group (13.3%). CONCLUSIONS AND CLINICAL IMPORTANCE: These results confirm previous studies and clinical transfusion medicine experience. Naturally occurring anti-DEA 7 alloantibodies have been reported but their clinical relevance has not been shown. Pregnancy does not appear to sensitize dogs to RBC antigens. Consequently, dogs with prior history of pregnancy can be used safely as blood donors. Conversely, no additional pretransfusion compatibility studies would be required should these dogs themselves need to be transfused.

Writing a Cochrane systematic review on preventive interventions to improve safety: the case of the construction industry.

OBJECTIVES: The objective of this paper is to describe the main steps and to conduct a systematic literature review on preventive interventions concerning work-related injuries and to illustrate the process. METHODS: Based on the Cochrane handbook, a structured framework of six steps was outlined for the development of a systematic review. This framework was used to describe a Cochrane systematic review (CSR) on the effectiveness of interventions to prevent work related injuries in the construction industry. RESULTS: The 6 main steps to write a CSR were: formulating the problem and objectives; locating and selecting studies; assessing study quality; collecting data; analysing data and presenting results; and interpreting results. The CSR on preventing injuries in the construction industry yielded five eligible intervention studies. Re-analysis of original injury data of the studies on regulatory interventions, through correcting for pre-intervention injury trends led to different conclusions about the effectiveness of interventions than those reported in the original studies. CONCLUSIONS: The Cochrane handbook for systematic reviews of interventions provides a practical and feasible six-step framework for developing and reporting a systematic review for preventive interventions.

The utility of NBS-profiling for characterization of yellow rust resistance in an F6 durum wheat population.

Seedling and adult plant (field) resistance to yellow rust in the durum wheat (Triticum turgidum ssp. durum) cross Kunduru-1149 x Cham-1 was characterized using a functionally-targeted DNA marker system, NBS-profiling. Chi-squared analysis indicated a four gene model conferring seedling yellow rust resistance against Puccinia striiformis f. sp. tritici isolate WYR85/22 (virulent on Yr2, Yr6, Yr7 and Yr9). Interval mapping located two QTL for yellow rust resistance on the long arm of chromosome 1B, while Kruskal-Wallis single marker regression identified a number of additional marker loci associated with seedling and/or adult plant, field resistance to yellow rust. These results suggested that much of the yellow rust resistance seen in the field may be due to seedling expressed resistance (R) genes. Characterization of the DNA sequence of three NBS marker loci indicated that all showed significant homology to functionally-characterized R-genes and resistance gene analogues (RGAs), with the greatest homology being NBS-LRR-type R-genes and RGAs from cereal species.

Genetic and functional association of FAM5C with myocardial infarction.

BACKGROUND: We previously identified a 40 Mb region of linkage on chromosome 1q in our early onset coronary artery disease (CAD) genome-wide linkage scan (GENECARD) with modest evidence for linkage (n = 420, LOD 0.95). When the data are stratified by acute coronary syndrome (ACS), this modest maximum in the overall group became a well-defined LOD peak (maximum LOD of 2.17, D1S1589/D1S518). This peak overlaps a recently identified inflammatory biomarker (MCP-1) linkage region from the Framingham Heart Study (maximum LOD of 4.27, D1S1589) and a region of linkage to metabolic syndrome from the IRAS study (maximum LOD of 2.59, D1S1589/D1S518). The overlap of genetic screens in independent data sets provides evidence for the existence of a gene or genes for CAD in this region. METHODS: A peak-wide association screen (457 SNPs) was conducted of a region 1 LOD score down from the peak marker (168-198 Mb) in a linkage peak for acute coronary syndrome (ACS) on chromosome 1, within a family-based early onset coronary artery disease (CAD) sample (GENECARD). RESULTS: Polymorphisms were identified within the 'family with sequence similarity 5, member C' gene (FAM5C) that show genetic linkage to and are associated with myocardial infarction (MI) in GENECARD. The association was confirmed in an independent CAD case-control sample (CATHGEN) and strong association with MI was identified with single nucleotide polymorphisms (SNPs) in the 3' end of FAM5C. FAM5C genotypes were also correlated with expression of the gene in human aorta. Expression levels of FAM5C decreased with increasing passage of proliferating aortic smooth muscle cells (SMC) suggesting a role for this molecule in smooth muscle cell proliferation and senescence. CONCLUSION: These data implicate FAM5C alleles in the risk of myocardial infarction and suggest further functional studies of FAM5C are required to identify the gene's contribution to atherosclerosis.

Diagnosis of acute hepatitis C virus infection and estimated incidence in low- and high-risk English populations.

The diagnosis of acute hepatitis C virus (HCV) infection is not straightforward; few people exhibit clinical symptoms and genome/antigen detection techniques do not indicate when infection had occurred. Here, a strategy to detect HCV RNA in the absence of antibody ('window-period') for diagnosis of acute infection is assessed. The sentinel surveillance of hepatitis testing study was used to retrospectively identify anti-HCV negative samples from high-risk individuals (2002-2003), for testing singly for HCV RNA. Additional samples were identified prospectively (2005) and tested in pools for HCV RNA. Positive samples were genotyped. Incidence and costs of adopting the pooling strategy were estimated. In the retrospective study, 8/390 (2.1%) samples were confirmed HCV RNA positive, anti-HCV negative. Prospectively, 3237 samples were tested in 325 pools. Five positive pools identified four confirmed HCV RNA positive patients (one false positive). Estimated incidence was 12.9 per 100 person-years in injecting drug users (IDUs) (retrospective study) and 3.7 per 100 person-years among drug/alcohol services and prison attendees (prospective study). Estimated costs were pound 850 per positive sample, in areas of higher risk. The yield from a window-period strategy depends upon the population tested. Pooled HCV RNA testing of anti-HCV negative samples from the current IDUs is realistic and relatively inexpensive to identify recently infected individuals.