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Abstract Combined action observation and motor imagery (AOMI) facilitates corticospinal excitability (CSE) and may potentially induce plastic-like changes in the brain in a similar manner to physical practice. This study used transcranial magnetic stimulation (TMS) to explore changes in CSE for AOMI of coordinative lower-limb actions. Twenty-four healthy adults completed two baseline (BLH, BLNH) and three AOMI conditions, where they observed a knee extension while simultaneously imagining the same action (AOMICONG), plantarflexion (AOMICOOR-FUNC), or dorsiflexion (AOMICOOR-MOVE). Motor evoked potential (MEP) amplitudes were recorded as a marker of CSE for all conditions from two knee extensor, one dorsi flexor, and two plantar flexor muscles following TMS to the right leg representation of the left primary motor cortex. A main effect for experimental condition was reported for all three muscle groups. MEP amplitudes were significantly greater in the AOMICONG condition compared to the BLNH condition (p = .04) for the knee extensors, AOMICOOR-FUNC condition compared to the BLH condition (p = .03) for the plantar flexors, and AOMICOOR-MOVE condition compared to the two baseline conditions for the dorsi flexors (ps ≤ .01). The study findings support the notion that changes in CSE are driven by the imagined actions during coordinative AOMI.

Abstract Background Platelets and erythrocytes constitute over 95% of all hematopoietic stem cell output. However, the clonal dynamics of HSC contribution to these lineages remains largely unexplored. Results We use lentiviral genetic labeling of mouse hematopoietic stem cells to quantify output from all lineages, nucleate, and anucleate, simultaneously linking these with stem and progenitor cell transcriptomic phenotypes using single-cell RNA-sequencing. We observe dynamic shifts of clonal behaviors through time in same-animal peripheral blood and demonstrate that acute platelet depletion shifts the output of multipotent hematopoietic stem cells to the exclusive production of platelets. Additionally, we observe the emergence of new myeloid-biased clones, which support short- and long-term production of blood cells. Conclusions Our approach enables kinetic studies of multi-lineage output in the peripheral blood and transcriptional heterogeneity of individual hematopoietic stem cells. Our results give a unique insight into hematopoietic stem cell reactivation upon platelet depletion and of clonal dynamics in both steady state and under stress.

Background Muscular forces drive proximal humeral fracture deformity, yet it is unknown if arm position can help mitigate such forces. Our hypothesis was that glenohumeral abduction and humeral internal rotation decrease the pull of the supraspinatus and subscapularis muscles, minimizing varus fracture deformity. Methods A medial wedge osteotomy was performed in eight cadaveric shoulders to simulate a two-part fracture. The specimens were tested on a custom shoulder testing system. Humeral head varus was measured following physiologic muscle loading at neutral and 20° humeral internal rotation at both 0° and 20° glenohumeral abduction. Results There was a significant decrease in varus deformity caused by the subscapularis (p

Trabectedin is a chemotherapeutic used to treat advanced soft tissue sarcoma and relapsed platinum-sensitive ovarian cancer. Although it is associated with a low incidence of cardiotoxicity, when this occurs it can be fatal or significantly compromise the quality of life in patients with advanced cancer. Here, we present a series of 4 cases where trabectedin-treated sarcoma patients developed cardiovascular complications. Similar to previous literature describing this association, all patients had prior treatment with anthracyclines and presented at different time points following treatment initiation. Each patient presented with exertional breathlessness and was found to have severely impaired left ventricular systolic function (ejection fraction ≤35%), and 1 patient had concurrent atrial fibrillation with a fast ventricular rate. All of the patients were treated with neurohormonal blockade, and a multi-disciplinary decision was made to stop trabectedin in 3 patients and continue in 1 patient. Two of the 4 patients had an improvement in their left ventricular systolic function. It is unclear what effect preceeding anthracycline or tyrosine kinase inhibitor treatment has in priming patients to develop cardiotoxicity in this setting. Our case series adds to the evidence surrounding this association and highlights that trabectedin-associated cardiotoxicity can present in an insidious fashion.

Research has shown that motor imagery (MI) interventions, involving the systematic and repeated imagination of visual and kinaesthetic components of movements, can enhance performance in sport. Twenty years ago, Holmes and Collins (2001) published the PETTLEP model as a framework to improve the delivery and outcome of MI interventions. The model outlined seven principles to be considered when designing effective imagery interventions: Physical, Environment, Task, Timing, Learning, Emotion, Perspective (i.e., PETTLEP). The incorporation of these principles within MI interventions was assumed to facilitate performance through the optimisation of a functional equivalence between the neurophysiological substrates of motor preparation and execution, and that of MI. Since its conception, this model has become a standard for many in the delivery of imagery interventions in sport and has been corroborated through research and practice. This 20-year review first covers the reach and impact of this influential model and the more recent empirical investigations related to PETTLEP. We then outline how PETTLEP-based imagery may be integrated with action observation to support an increasingly popular approach to the delivery of imagery interventions in sport. Research indicates that combining these two simulation states can enhance sport performance whilst also providing the sport psychologist more control over the imagery experience than is possible through traditional imagery interventions. This article discusses the application of PETTLEP within a combined action observation and imagery framework and provides guidance for sport psychologists for the creation of new PETTLEP-informed interventions.

This paper covers three datasets containing texts in Ancient Greek, manually aligned at word level against translations in English (Grc-Eng), Portuguese (Grc-Por) and Latin (Grc-Lat). The datasets were collected by two domain experts through annotation on the Ugarit Translation Alignment Editor (https://ugarit.ialigner.com/). The quality of each dataset was measured through Inter-Annotator-Agreement (IAA) above 80%. Each dataset contains the aligned pairs and an Annotation Style Guide, and serves as a Gold Standard for translation alignment of Ancient Greek, for the evaluation of automatic translation alignment models, and as high-quality training data. The Annotation Style Guide provides a starting point to approach the task of translation alignment for research and teaching. The data is stored on GitHub and Zenodo.

Abstract Background Alternative splicing is a key mechanism underlying cellular differentiation and a driver of complexity in mammalian neuronal tissues. However, understanding of which isoforms are differentially used or expressed and how this affects cellular differentiation remains unclear. Long read sequencing allows full-length transcript recovery and quantification, enabling transcript-level analysis of alternative splicing processes and how these change with cell state. Here, we utilise Oxford Nanopore Technologies sequencing to produce a custom annotation of a well-studied human neuroblastoma cell line SH-SY5Y, and to characterise isoform expression and usage across differentiation. Results We identify many previously unannotated features, including a novel transcript of the voltage-gated calcium channel subunit gene, CACNA2D2. We show differential expression and usage of transcripts during differentiation identifying candidates for future research into state change regulation. Conclusions Our work highlights the potential of long read sequencing to uncover previously unknown transcript diversity and mechanisms influencing alternative splicing.

Cardiac implantable electronic device (CIED) implants are rising in an older, more co-morbid population. The prevalence of CIED infection ranges from 1–4%. Whilst complete extraction of all transvenous hardware is recommended for infected, eroded, or pre-eroding CIEDs, this approach is not without risk and may be unacceptable to some patients. Long-term data on a more conservative strategy is lacking. We report on our experience of conservative management with pocket revision as a primary strategy in carefully selected patients. Method: A retrospective review of all CIED revision procedures was undertaken at a large tertiary center, over a 7-year period, with a mean follow-up timeframe of 39 months. Results: A total of 86 patients underwent 96 revision procedures; 7 patients required further revisions and 13 went on to undergo CIED extraction by the end of the follow-up period. The overall rate of mortality at 12 months was 8.1%, increasing to 24.4% at the end of the follow-up period. Conclusion: Our data provide important outcome information on an alternative strategy to lead extraction in carefully selected patients where the risk of extraction is perceived to be unacceptable. The absence of systemic infection appears to predict better outcomes than previously reported, and over two-thirds of patients remained complication-free at 12 months.

Developmental coordination disorder (DCD) is characterised by poor motor coordination, which interferes with the ability to execute activities of daily living (ADLs). Combined action observation and motor imagery (AOMI) involves observing movement videos whilst imagining simultaneously the sensations of executing the same movement. Laboratory-based research indicates that AOMI can help improve movement coordination in children with DCD, but no previous research had investigated the efficacy of AOMI interventions for learning ADLs. This study investigated the efficacy of a home-based, parent-led, AOMI intervention for learning ADLs in children with DCD. Children with confirmed (n = 23) or suspected (n = 5) DCD (total sample n = 28), aged 7–12 years, were assigned to either an AOMI intervention or a control intervention (both n = 14). Participants attempted the following ADLs at pre-test (week 1), post-test (week 4), and retention test (week 6): shoelace tying, cutlery use, shirt buttoning, and cup stacking. Task completion times and movement techniques were recorded. The AOMI intervention produced significantly faster task completion times than the control intervention at post-test for shoelace tying, and significantly improved movement techniques for shoelace tying and cup stacking. Importantly, for children who could not tie shoelaces at pre-test (n = 9 per group), 89% of those following the AOMI intervention learnt the skill successfully by the end of the study, compared to only 44% of those following the control intervention. The findings indicate that home-based, parent-led, AOMI interventions can aid the learning of complex ADLs in children with DCD, and may be particularly effective for facilitating the learning of motor skills that do not currently exist within these children’s motor repertoire.

Respiratory viruses such as influenza do not typically cause viremia; however, SARS-CoV-2 has been detected in the blood of COVID-19 patients with mild and severe symptoms. Detection of SARS-CoV-2 in blood raises questions about its role in pathogenesis as well as transfusion safety concerns. Blood donor reports of symptoms or a diagnosis of COVID-19 after donation (post-donation information, PDI) preceded or coincided with increased general population COVID-19 mortality. Plasma samples from 2,250 blood donors who reported possible COVID-19–related PDI were tested for the presence of SARS-CoV-2 RNA. Detection of RNAemia peaked at 9%–15% of PDI donors in late 2020 to early 2021 and fell to approximately 4% after implementation of widespread vaccination in the population. RNAemic donors were 1.2- to 1.4-fold more likely to report cough or shortness of breath and 1.8-fold more likely to report change in taste or smell compared with infected donors without detectable RNAemia. No infectious virus was detected in plasma from RNAemic donors; inoculation of permissive cell lines produced less than 0.7–7 plaque-forming units (PFU)/mL and in susceptible mice less than 100 PFU/mL in RNA-positive plasma based on limits of detection in these models. These findings suggest that blood transfusions are highly unlikely to transmit SARS-CoV-2 infection.

Wright et al. report the first structure of a human TRPC1/4/5 channel in complex with a xanthine-based TRPC5 inhibitor Pico145. They find that Pico145 binds to a conserved lipid binding site of TRPC5, where it displaces a phospholipid. This study provides insights into the mechanism-of-action of xanthine-based TRPC1/4/5 modulators.

PGx testing requires a complex set of activities undertaken by practitioners and patients, resulting in varying implementation success. This systematic review aimed (PROSPERO: CRD42019150940) to identify barriers and enablers to practitioners and patients implementing pharmacogenomic testing. We followed PRISMA guidelines to conduct and report this review. Medline, EMBASE, CINAHL, PsycINFO, and PubMed Central were systematically searched from inception to June 2022. The theoretical domain framework (TDF) guided the organisation and reporting of barriers or enablers relating to pharmacogenomic testing activities. From the twenty-five eligible reports, eleven activities were described relating to four implementation stages: ordering, facilitating, interpreting, and applying pharmacogenomic testing. Four themes were identified across the implementation stages: IT infrastructure, effort, rewards, and unknown territory. Barriers were most consistently mapped to TDF domains: memory, attention and decision-making processes, environmental context and resources, and belief about consequences.

Within a single jurisdiction, electricity regulators may set varying tariff structures depending on the type of customer, to recover the cost of generating and delivering electricity. From this cost perspective, the various tariff structures are similar to one another. This paper however, demonstrates that the tariff structures are very different from the point of view of determining the economics of behind-the-meter solar microgrids. Based on empirical data for Ontario we analyse three tariff structures: time-of-use (ToU), customer peak demand charges (DC) and system-wide peak charges (PC). The operation of the microgrid is first optimized and then the internal rate of return (IRR) of the system over its lifetime is maximized by choosing the optimal battery size. This analysis is projected into the future for project start dates 2019–2025 and results in IRRs of 2.7%–7.7% (ToU, residential) 5.6%–13.4% (DC, day/evening use commercial building), 7.5%–17.6% (DC, day-use commercial building). The PC tariff structure was determined to be unsuited to a solar microgrid partly because trends in the system peak hours have resulted in 9.2% per year decline in solar energy yield since its introduction in 2010.

Abstract Background Deprescribing is a partnership between practitioners, patients and caregivers. External characteristics including age, comorbidities and polypharmacy are poor predictors of attitude towards deprescribing. This hospital-based study aimed to describe the desire of patients and caregivers to be involved in medicine decision-making, and identify attitudinal predictors of desire to try stopping a medicine. Methods Patients and caregivers recruited from seven Older People’s Medicine wards across two UK hospitals completed the revised Patients’Attitudes Towards Deprescribing (rPATD) questionnaire. Patients prescribed polypharmacy and caregivers involved in medication decision-making of such patients were eligible. A target of 150 patients and caregivers provided a 95% confidence interval of ±11.0% or smaller around rPATD item agreement. Descriptive statistics characterised participants and rPATD responses. Responses to items regarding desire to be involved in medication decision-making and desire to try stopping a medicine were used to address the aims. Binary logistic regression provided the adjusted odds ratios (OR) for predictors of desire to try stopping a medicine. Results Patient participants (N = 75) were a median (IQ) 87.0 (83.0, 90.0) years old and the median (IQ) number of pre-admission medication was 8.0 (6.0, 10.0). Caregiver participants (N = 76) were a median (IQ) 70.0 (57.0, 83.0) years old and the majority were a spouse (63.2%). For patients and caregivers respectively, the following were reported: 58.7 and 65.8% wanted to be involved in medication decision-making; 29.3 and 43.5% reported a desire to try stopping a medicine. Attitudinal predictors of low desire to try stopping a medicine for patients and caregivers are a perception that there are no unnecessary prescribed medicines [OR = 0.179 (patients) and 0.044 (caregivers)] and no desire for dose reduction [OR = 0.199 (patients) and 0.024 (caregivers)]. A perception of not being prescribed too many medicines also predicted low patient desire to try stopping a medicine [OR = 0.195]. Conclusion A substantial proportion of patients and caregivers did not want to be involved medication decision-making, however this should not result in practitioners dismissing deprescribing opportunities. The three diagnostic indicators for establishing desire to try stopping a medicine are perceived necessity of the medicine, appropriateness of the number prescribed medications and a desire for dose reduction.

Abstract Molecular methods have greatly increased our understanding of the previously cryptic spatial ecology of bumble bees (Bombus spp.), with knowledge of the spatial ecology of these bees being central to conserving their essential pollination services. Bombus hypnorum, the Tree Bumble Bee, is unusual in that it has recently rapidly expanded its range, having colonized much of the UK mainland since 2001. However, the spatial ecology of B. hypnorum has not previously been investigated. To address this issue, and to investigate whether specific features of the spatial ecology of B. hypnorum are associated with its rapid range expansion, we used 14 microsatellite markers to estimate worker foraging distance, nest density, between‐year lineage survival rate and isolation by distance in a representative UK B. hypnorum population. After assigning workers to colonies based on full or half sibship, we estimated the mean colony‐specific worker foraging distance as 103.6 m, considerably less than values reported from most other bumble bee populations. Estimated nest density was notably high (2.56 and 0.72 colonies ha−1 in 2014 and 2015, respectively), estimated between‐year lineage survival rate was 0.07, and there was no evidence of fine‐scale isolation by distance. In addition, genotyping stored sperm dissected from sampled queens confirmed polyandry in this population (mean minimum mating frequency of 1.7 males per queen). Overall, our findings establish critical spatial ecological parameters and the mating system of this unusual bumble bee population and suggest that short worker foraging distances and high nest densities are associated with its rapid range expansion.

Ankle fractures are one of the most common injuries treated by orthopedic surgeons worldwide. However, operative indications, techniques, and reported outcomes following operative fixation vary widely in the literature. This evidence-based review focuses on recent advances in the operative management of ankle fractures including arthroscopic-assisted surgery, deltoid ligament complex repair, expanded indications for posterior malleolus fixation, fibula intramedullary nailing, and dynamic syndesmosis repair. Level of Evidence: Level V, expert opinion.

Background: After home health care, the skilled nursing facility (SNF) is the most commonly used postacute care modality, among Medicare beneficiaries, after total joint arthroplasty. Prior studies demonstrated that a loss in postsurgical ambulatory gains is incurred in the interval between hospital discharge and arrival at the SNF. The aim of this present study is to determine the consequences of that loss in function, as well as compare SNF-related outcomes in patients with Medicare vs Managed Care (MC) insurance. Methods: We conducted a retrospective analysis of 80 patients (54 Medicare and 26 MC) who attended an SNF after hospitalization for total joint arthroplasty. Outcomes from physical therapy records were abstracted from each patient's SNF file. Results: There was an approximately 40% drop-off in gait achievements between hospital discharge and SNF admission. This decline in ambulation was significantly greater in Medicare patients (Medicare: 94.6 ± 123.2 ft, MC: 40.0 ± 48.9 ft, P = .034). Larger reductions in gait achievements between hospital discharge and SNF admission were significantly correlated with longer SNF lengths of stay and poorer gait achievements by SNF discharge. Patients with MC insurance made significant improvements in gait training at the SNF beyond that which was acquired at the hospital, whereas Medicare patients did not (PMedicare = .28, PMC = .003). Conclusions: Large losses in motor function between hospital discharge and SNF admission were associated with poor functional outcomes and longer stays at the SNF. These effects were more pronounced in Medicare patients than those with MC insurance. Keywords: Total joint arthroplasty, Skilled nursing facility, Medicare, Managed Care, Physical therapy

Community pharmacy services have evolved to include medical and pharmaceutical interventions alongside dispensing. While established pharmacogenomic (PGx) testing is available throughout the Netherlands, this is primarily based in hospital environments and for specialist medicines. The aim of this work was to describe how best to implement PGx services within community pharmacy, considering potential barriers and enablers to service delivery and how to address them. The service was implemented across a selection of community pharmacies in the Netherlands. Data were captured on test outcomes and through a pharmacist survey. Following testing, 17.8% of the clinical samples were recommended to avoid certain medication (based on their current medicines use), and 14.0% to have their dose adjusted. Pre-emptive analysis of genotyped patients showed that the majority (99.2%) had actionable variants. Pharmacists felt confident in their operational knowledge to deliver the service, but less so in applying that knowledge. Delivering the service was believed to improve relationships with other healthcare professionals. These results add to the evidence in understanding how PGx can be delivered effectively within the community pharmacy environment. Training pharmacists in how to respond to patient queries and make clinical recommendations may enhance service provision further.

Dysphagia is common—not only associated with stroke, dementia, Parkinson’s but also in many non-neurological medical problems—and is increasingly prevalent in ageing patients, where malnutrition is common and pneumonia is frequently the main cause of death. To improve the care of people with dysphagia (PWD) and minimise risk of aspiration and choking, the textures of food and drinks are frequently modified. Whilst medicines are usually concurrently prescribed for PWD, their texture is frequently not considered and therefore any minimisation of risk with respect to food and drink may be being negated when such medicines are administered. Furthermore, evidence is starting to emerge that mixing thickeners with medicines can, in certain circumstances, significantly affect drug bioavailability and therefore amending the texture of a medicine may not be straightforward. Research across a number of hospital trusts demonstrated that PWD are three times more likely to experience medication administration errors than those without dysphagia located on the same ward. Errors more commonly seen in PWD were missed doses, wrong formulation and wrong preparation through medicines alteration. Researchers also found that the same patient with dysphagia would be given their medicines in entirely different ways depending on the person administering the medicine. The alteration of medicines prior to administration has potential for patient harm, particularly if the medicine has been designed to release medicines at a pre-defined rate or within a pre-defined location. Alteration of medicines can have significant legal implications and these are frequently overlooked. Dispersing, crushing or mixing medicines can be part of, or misconstrued as, covert administration, thus introducing a further raft of legislation. Guidance within the UK recommends that following identification of dysphagia, the ongoing need for the medicine should be considered, as should the most appropriate route and formulation, with medicines alteration used as a last resort. The patient should be at the centre of any decision making. Evidence suggests that in the UK this guidance is not being followed. This article considers the clinical and legal issues surrounding administration of medicines to PWD from a UK perspective and debates whether medicines optimisation should be the primary responsibility of the prescriber when initiating therapy on the ward or the nurse who administers the medicine.

Micro-tracked CPV, in which cells move relative to fixed concentrating optics, allows CPV to be deployed in the same manner as fixed PV modules. Behind-the-meter applications in locations where there is a land/roof area cost for the space occupied by the modules confers a cost advantage to CPV compared to PV. The primary objective of the present paper is to estimate target prices below which CPV has a competitive advantage over PV. We analyse PV and CPV microgrids, optimizing the scheduling of power into and out of the battery in order to achieve the maximum savings compared to purchasing grid power. We then choose the battery capacity that maximizes the internal rate of return (IRR) on the PV microgrid. The CPV target price is the price that gives a CPV IRR that matches the PV IRR. The target CPV prices obtained are found to depend on the land price and on the ratio of CPV to PV annual energy yield (REY), but are insensitive to battery prices, load profiles, current electricity tariffs and future trends in electricity tariffs. Modesto, CA has a REY of 1.10 and an average target CPV price from $1.86/W at zero land price to $3.53/W at $400/m2. Lancaster, CA with a REY of 1.26 has corresponding target CPV prices from $2.14 to $4.23/W. In Las Vegas, NV, (REY = 1.27) the target CPV price is $2.13/W at a land price of zero, but at higher land prices the IRR is insufficient for solar power to be deployed. These target CPV prices correspond with current estimates of pedestal-tracked CPV of $2.4 to $3.3/W and indicate that it is realistic to expect micro-tracked CPV to be cost competitive with PV in some urban areas in southwestern USA.

Action observation has been suggested to be an effective adjunct to physical practice in motor (re)learning settings. However, optimal viewing conditions for interventions are yet to be established. Single-pulse transcranial magnetic stimulation (TMS) was used to investigate the effect of two different screen positions and participants’ screen position viewing preference on the amplitude of motor evoked potentials (MEPs) during observation of a ball pinch action. Twenty-four participants observed four blocked conditions that contained either a dynamic index finger-thumb ball pinch or a static hand holding a ball in a similar position on a horizontally or vertically positioned screen. TMS was delivered to the hand representation of the left primary motor cortex and MEPs were recorded from the first dorsal interosseous muscle of the right hand. Initial analysis of the normalized MEP amplitude data showed no significant differences between conditions. In a follow-up procedure, participants engaged in individual semi-structured interviews and completed a questionnaire designed to assess viewing affect and screen position viewing preference. The MEP data were subsequently split by screen position preference and re-analyzed using a 2 × 2 repeated measures ANOVA. Main effects indicated that participants who preferred the horizontal screen position (n = 16) demonstrated significantly greater MEP amplitudes during observation of the ball-pinch action compared to the static hand condition irrespective of screen position, and during the horizontal compared to the vertical screen position irrespective of video type. These results suggest that ensuring anatomical and perceptual congruency with the physical task, alongside consideration of participants’ screen position viewing preferences, may be an important part of optimizing action observation interventions.

Research in care homes has demonstrated that medication management practices in patients with dysphagia and those receiving medicines covertly may be inappropriate, illegal, and potentially cause harm. This paper presents the results of a feasibility study piloting a resident and healthcare professional best practice charter to improve such practices in care home residents with dysphagia. A charter was developed through a multi-professional expert panel, implemented in one care home, and then piloted in 22 homes in England, Wales, and Northern Ireland. A website was setup and developed iteratively to support the process. Care home staff and residents provided initial feedback on the implementation process and on perceived outcomes six months post implementation. A total of 16 (88.9%) out of 18 respondents from nine homes for six months reported a positive response to the charter. More than 80% of responses regarding the implementation process, impact on staff confidence, and perceived usefulness of the charter were positive. Perceived effectiveness and usefulness could, however, be further improved especially the perceived effect on frequency of medication review, which is reliant on external stakeholder involvement. The charter and supporting website were well received with respondents believing that it was useful, staff showing more confidence, and residents having enhanced care. Approaches to enhancing the charter’s effectiveness were identified.

During the 2012 outbreak of West Nile virus in the United States, approximately one third of the cases were in Texas. Of those, about half occurred in northern Texas. Models based on infected blood donors and persons with neuroinvasive disease showed, respectively, that ≈0.72% and 1.98% of persons in northern Texas became infected.

Proteins of the TRPC family can form many homo- and heterotetrameric cation channels permeable to Na+, K+ and Ca2+. In this review, we focus on channels formed by the isoforms TRPC1, TRPC4 and TRPC5. We review evidence for the formation of different TRPC1/4/5 tetramers, give an overview of recently developed small-molecule TRPC1/4/5 activators and inhibitors, highlight examples of biological roles of TRPC1/4/5 channels in different tissues and pathologies, and discuss how high-quality chemical probes of TRPC1/4/5 modulators can be used to understand the involvement of TRPC1/4/5 channels in physiological and pathophysiological processes.

National blood donor screening for West Nile virus (WNV) RNA using minipool nucleic acid amplification testing (MP-NAT) was implemented in the United States in July 2003. We compiled national NAT yield data and performed WNV immunoglobulin M (IgM) testing in 1 WNV-epidemic region (North Dakota). State-specific MP-NAT yield, antibody seroprevalence, and the average time RNA is detectable by MP-NAT were used to estimate incident infections in 2003. WNV donor screening yielded 944 confirmed viremic donors. MP-NAT yield peaked in August with >0.5% of donations positive for WNV RNA in 4 states. Peak IgM seroprevalence for North Dakota was 5.2% in late September. The average time viremia is detectable by MP-NAT was 6.9 days (95% confidence interval [CI] 3.0–10.7). An estimated 735,000 (95% CI 322,000–1,147,000) infections occurred in 2003, with 256 (95% CI 112–401) infections per neuroinvasive case. In addition to preventing transfusion-transmitted WNV infection, donor screening can serve as a tool to monitor seasonal incidence in the general population.

An automated high-throughput method that employs rolling circle amplification (RCA) to generate template for large-scale DNA sequencing has been developed using liquid handling systems equipped with precision glass syringes. A protocol was designed to perform the sequencing analysis from template preparation to thermal cycle sequencing within the same vessel, thus minimizing the amount of liquid handling and transfer. The amplified DNA was directly used for cycle sequencing with no need for any purification procedures. Total RCA reaction volumes as low as 500 nL generated sufficient templates for successful sequencing. Reducing the RCA total reaction volumes by a 40-fold factor, from a total of 20 μL to 500 nL, resulted in a significant reduction in cost, from $1.25/reaction to less than $0.04/reaction. Additionally, the volume of the sequencing reactions was reduced from a total of 20 to 10 μL, thus generating a further cost advantage. This high-throughput DNA sequencing protocol maximizes the speed and precision of processing while significantly reducing the cost of amplification.

Dietary iron absorption is regulated by hepcidin, an iron regulatory protein produced by the liver. Hepcidin production is regulated by iron stores, erythropoiesis and inflammation, but its physiology when repeated blood loss occurs has not been characterized. Hepcidin was assayed in plasma samples obtained from 114 first-time/reactivated (no blood donations in preceding 2 years) female donors and 34 frequent (≥3 red blood cell donations in preceding 12 months) male donors as they were phlebotomized ≥4 times over 18–24 months. Hepcidin levels were compared to ferritin and hemoglobin levels using multivariable repeated measures regression models. Hepcidin, ferritin and hemoglobin levels declined with increasing frequency of donation in the first-time/reactivated females. Hepcidin and ferritin levels correlated well with each other (Spearman’s correlation of 0.74), but on average hepcidin varied more between donations for a given donor relative to ferritin. In a multivariable repeated measures regression model the predicted inter-donation decline in hemoglobin varied as a function of hepcidin and ferritin; hemoglobin was 0.51 g/dL lower for subjects with low (>45.7 ng/mL) or decreasing hepcidin and low ferritin (>26 ng/mL), and was essentially zero for other subjects including those with high (>45.7 ng/mL) or increasing hepcidin and low ferritin (>26 ng/mL) levels (P