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The Ewald method is applied to accelerate the evaluation of the Green's function (GF) of an infinite equispaced linear array of point sources with linear phasing. Only a few terms are needed to evaluate Ewald sums, which are cast in terms of error functions and exponential integrals, to high accuracy. It is shown analytically that the choice of the standard "optimal" Ewald splitting parameter E0 causes overflow errors at high frequencies (period large compared to the wavelength), and convergence rates are analyzed. A recipe for selecting the Ewald splitting parameter is provided. © 2006 Elsevier Inc. All rights reserved.

The Ewald method is applied to accelerate the evaluation of the Green's function of an infinite periodic phased array of line sources. The Ewald representation for a cylindrical wave is obtained from the known representation for the spherical wave, and a systematic general procedure is applied to extend previous results. Only a few terms are needed to evaluate Ewald sums, which are cast in terms of error functions and exponential integrals, to high accuracy. Singularities and convergence rates are analyzed, and a recipe for selecting the Ewald splitting parameter ε is given to handle both low and high frequency ranges. Indeed, it is shown analytically that the choice of the standard optimal splitting parameter ε0 will cause overflow errors at high frequencies. Numerical examples illustrate the results and the sensitivity of the Ewald representation to the splitting parameter ε. © 2005 IEEE.

An alternative direct procedure for applying the Ewald approach to obtain the Green's function for an array of line sources with 1D periodicity is presented. Furthermore, an algorithm for choosing the Ewald splitting parameter E that extends the efficiency of the method when the periodicity is somewhat larger than a wavelength is derived. In particular, the dyadic Green's function formalism is combined with the Ewald method.

Background A novel handheld point-of-care high-sensitivity cardiac troponin I analyser has recently been introduced to the market. Evaluating its diagnostic performance against laboratory standards is imperative, given the variations in cardiac troponin levels across populations. This study compared the diagnostic performance between the point-of-care high-sensitivity cardiac troponin I assay (Siemens Healthineers Atellica VTLi) and a laboratory high-sensitivity cardiac troponin I assay (Abbott ARCHITECT STAT High Sensitive Troponin-I) performed using blood samples from various populations (overall, male, female, younger and older) of Chinese patients with chest pain.Methods This cross-sectional study included 585 consecutive Chinese patients (age ≥18 year) who presented to an emergency department with chest pain (lasting >5 min) and were managed following the chest pain protocol between 1 August 2023 and 12 June 2024. For both assays, blood samples were collected at two time points (0 hour (initial) and 3 hour (subsequent)). The primary outcome was the diagnostic performance of the two assays, evaluated with their 99th percentile upper reference limits used as the cut-off values for diagnosing myocardial infarction. The gold standard for comparison was the final diagnoses made by attending physicians.Results The point-of-care and laboratory assays exhibited equivalent sensitivity and negative predictive values (both 100%) for blood samples collected at both time points. However, the point-of-care assay outperformed the laboratory assay in terms of specificity (initial: 90.5% to 96.3% vs 79.8% to 94.7%; subsequent: 87.8% to 94.8% vs 77.7% to 92.4%) and positive predictive value (initial: 24.4% to 30.8% vs 11.6% to 23.5%; subsequent: 12.5% to 25.0% vs 5.9% to 18.8%), particularly in older patients.Conclusion The point-of-care assay is recommended for rapid clinical decision-making. Future studies should explore the effects of its integration into clinical practice and the feasibility of using sex–race–age-specific 99th percentile upper reference limits to enhance its diagnostic performance.

Rice (Oryza sativa) is one of the most important cereal crops with a diverse set of pests and natural enemies. Rice fields often support a high diversity of arthropods which contribute significantly to productivity. This diversity is frequently threatened due to indiscriminate applications of pesticides. Our aim was to emphasize on the predator diversity in agrochemical exposed rice field as well as on the impact of surrounding vegetation on beneficial insect diversity. Natural enemies’ data were recorded from randomly selected 10 quadrates by visual observation from each treatment. A total of 5,590 individuals of predators were observed during the study period which included 27 species belonging to 16 families from five orders of arthropods during the kharif season of rice. Statistically, there were no significant differences between the population of general natural enemies such as Odonata, Coleoptera, Hymenoptera, and Araneae in plots with insecticide and control during the different growth stages of rice cultivation. Diversity indices were almost similar in fields where pesticide was sprayed and not sprayed. Our study concluded that natural enemies are conserved by ensuring crop heterogeneity, growing insect-friendly plants (with high levels of nectar and pollen) as border crops, and judicious application of granule insecticide like cartap hydrochloride in a rice agro-ecosystem.

Substance use and misuse among racial/ethnic subgroups remains a much understudied area. This article summarizes recent data from national surveys (SAMHSA) recording prevalence and treatment rates for major ethnic/racial subgroups. The authors point to a concern about the higher prevalence and treatment rates of alcohol among older American Indians and Alaskan Natives, but warn that more precise estimates must be obtained. They also address the finding that older African Americans have higher prevalence and rates of treatment admissions for certain illicit drugs such as cocaine. They highlight the unique concerns for older African Americans as the largest minority group within the U. S. Finally, a research agenda for the twenty-first century is suggested which will stress emphasis on continued, careful analysis of the life course of substance use among racial/ethnic subgroups. [ABSTRACT FROM AUTHOR]

Interest in performance-enhancing supplements is growing among younger athletes despite serious unresolved questions about safety and efficacy. Here's the information you need to talk to your patients about supplements they may be taking. [ABSTRACT FROM AUTHOR]

A series of N-acetylated, methionine-containing dipeptides designated AcMet-Aa containing various C-terminal amino acids designated Aa are hydrolyzed in aqueous solution at 50 ºC and 0.95 < pD < 1.10 in the presence of three cis-[Pd(L)(H2O)2]2+ complexes, in which L are bidentate ligands en Me4en, and 3-OH-dtco. The reactions were monitored by 1H-NMR spectroscopy. The rate constant for hydrolytic cleavage of the Met-Aa bond decreases as the steric bulk of the amino acid Aa increases. Correlations to Taft’s Es values were made. The substituents on α-C and β-C atoms lower the rate constant most, those on the Ё-C atom lower it less, and those on the δ-C have no detectable effect. Partial selectivity for leaving amino acid Aa is attributed to differences in the volume of the side chain and to discrimination between leaving groups of similar volume but different branching patterns.

Abstract The recent rise in speed and efficiency of new sequencing technologies have facilitated high-throughput sequencing, assembly and analyses of genomes, advancing ongoing efforts to analyze genetic sequences across major vertebrate groups. Standardized procedures in acquiring high quality DNA and RNA and establishing cell lines from target species will facilitate these initiatives. We provide a legal and methodological guide according to four standards of acquiring and storing tissue for the Genome 10K Project and similar initiatives as follows: four-star (banked tissue/cell cultures, RNA from multiple types of tissue for transcriptomes, and sufficient flash-frozen tissue for 1 mg of DNA, all from a single individual); three-star (RNA as above and frozen tissue for 1 mg of DNA); two-star (frozen tissue for at least 700 μg of DNA); and one-star (ethanol-preserved tissue for 700 μg of DNA or less of mixed quality). At a minimum, all tissues collected for the Genome 10K and other genomic projects should consider each species’ natural history and follow institutional and legal requirements. Associated documentation should detail as much information as possible about provenance to ensure representative sampling and subsequent sequencing. Hopefully, the procedures outlined here will not only encourage success in the Genome 10K Project but also inspire the adaptation of standards by other genomic projects, including those involving other biota.

Abstract Background Matrix extracellular phosphoglycoprotein (MEPE) belongs to a family of small integrin-binding ligand N-linked glycoproteins (SIBLINGs) that play a key role in skeleton development, particularly in mineralization, phosphate regulation and osteogenesis. MEPE associated disorders cause various physiological effects, such as loss of bone mass, tumors and disruption of renal function (hypophosphatemia). The study of this developmental gene from an evolutionary perspective could provide valuable insights on the adaptive diversification of morphological phenotypes in vertebrates. Results Here we studied the adaptive evolution of the MEPE gene in 26 Eutherian mammals and three birds. The comparative genomic analyses revealed a high degree of evolutionary conservation of some coding and non-coding regions of the MEPE gene across mammals indicating a possible regulatory or functional role likely related with mineralization and/or phosphate regulation. However, the majority of the coding region had a fast evolutionary rate, particularly within the largest exon (1467 bp). Rodentia and Scandentia had distinct substitution rates with an increased accumulation of both synonymous and non-synonymous mutations compared with other mammalian lineages. Characteristics of the gene (e.g. biochemical, evolutionary rate, and intronic conservation) differed greatly among lineages of the eight mammalian orders. We identified 20 sites with significant positive selection signatures (codon and protein level) outside the main regulatory motifs (dentonin and ASARM) suggestive of an adaptive role. Conversely, we find three sites under selection in the signal peptide and one in the ASARM motif that were supported by at least one selection model. The MEPE protein tends to accumulate amino acids promoting disorder and potential phosphorylation targets. Conclusion MEPE shows a high number of selection signatures, revealing the crucial role of positive selection in the evolution of this SIBLING member. The selection signatures were found mainly outside the functional motifs, reinforcing the idea that other regions outside the dentonin and the ASARM might be crucial for the function of the protein and future studies should be undertaken to understand its importance.

Abstract Background Cytosolic glutathione transferases (cGST) are a large group of ubiquitous enzymes involved in detoxification and are well known for their undesired side effects during chemotherapy. In this work we have performed thorough phylogenetic analyses to understand the various aspects of the evolution and functional diversification of cGSTs. Furthermore, we assessed plausible correlations between gene duplication and substrate specificity of gene paralogs in humans and selected species, notably in mammalian enzymes and their natural substrates. Results We present a molecular phylogeny of cytosolic GSTs that shows that several classes of cGSTs are more ubiquitous and thus have an older ancestry than previously thought. Furthermore, we found that positive selection is implicated in the diversification of cGSTs. The number of duplicate genes per class is generally higher for groups of enzymes that metabolize products of oxidative damage. Conclusions 1) Protection against oxidative stress seems to be the major driver of positive selection in mammalian cGSTs, explaining the overall expansion pattern of this subfamily; 2) Given the functional redundancy of GSTs that metabolize xenobiotic chemicals, we would expect the loss of gene duplicates, but by contrast we observed a gene expansion of this family, which likely has been favored by: i) the diversification of endogenous substrates; ii) differential tissue expression; and iii) increased specificity for a particular molecule; 3) The increased availability of sequence data from diversified taxa is likely to continue to improve our understanding of the early origin of the different cGST classes.

Abstract Background The mitochondria produce up to 95% of a eukaryotic cell's energy through oxidative phosphorylation. The proteins involved in this vital process are under high functional constraints. However, metabolic requirements vary across species, potentially modifying selective pressures. We evaluate the adaptive evolution of 12 protein-coding mitochondrial genes in 41 placental mammalian species by assessing amino acid sequence variation and exploring the functional implications of observed variation in secondary and tertiary protein structures. Results Wide variation in the properties of amino acids were observed at functionally important regions of cytochrome b in species with more-specialized metabolic requirements (such as adaptation to low energy diet or large body size, such as in elephant, dugong, sloth, and pangolin, and adaptation to unusual oxygen requirements, for example diving in cetaceans, flying in bats, and living at high altitudes in alpacas). Signatures of adaptive variation in the NADH dehydrogenase complex were restricted to the loop regions of the transmembrane units which likely function as protons pumps. Evidence of adaptive variation in the cytochrome c oxidase complex was observed mostly at the interface between the mitochondrial and nuclear-encoded subunits, perhaps evidence of co-evolution. The ATP8 subunit, which has an important role in the assembly of F0, exhibited the highest signal of adaptive variation. ATP6, which has an essential role in rotor performance, showed a high adaptive variation in predicted loop areas. Conclusion Our study provides insight into the adaptive evolution of the mtDNA genome in mammals and its implications for the molecular mechanism of oxidative phosphorylation. We present a framework for future experimental characterization of the impact of specific mutations in the function, physiology, and interactions of the mtDNA encoded proteins involved in oxidative phosphorylation.

Abstract Background Insensitivity of advanced-stage prostate cancer to androgen ablation therapy is a serious problem in clinical practice because it is associated with aggressive progression and poor prognosis. Targeted therapeutic drug discovery efforts are thwarted by lack of adequate knowledge of gene(s) associated with prostate tumorigenesis. Therefore there is the need for studies to provide leads to targeted intervention measures. Here we propose that stable expression of U94, a tumor suppressor gene encoded by human herpesvirus 6A (HHV-6A), could alter gene expression and thereby inhibit the tumorigenicity of PC3 cell line. Microarray gene expression profiling on U94 recombinant PC3 cell line could reveal genes that would elucidate prostate cancer biology, and hopefully identify potential therapeutic targets. Results We have shown that stable expression of U94 gene in PC3 cell line inhibited its focus formation in culture, and tumorigenesis in nude mice. Moreover gene expression profiling revealed dramatic upregulation of FN 1 (fibronectin, 91 ± 16-fold), and profound downregulation of ANGPTL 4 (angiopoietin-like-4, 20 ± 4-fold) in U94 recombinant PC3 cell line. Quantitative real-time polymerase chain reaction (QRT-PCR) analysis showed that the pattern of expression of FN 1 and ANGPTL 4 mRNA were consistent with the microarray data. Based on previous reports, the findings in this study implicate upregulation of FN 1 and downregulation of ANGPTL 4 in the anti tumor activity of U94. Genes with cancer inhibitory activities that were also upregulated include SERPINE 2 (serine/cysteine protease inhibitor 2, 7 ± 1-fold increase) and ADAMTS 1 (a disintegrin-like and metalloprotease with thrombospondin type 1 motif, 7 ± 2-fold increase). Additionally, SPUVE 23 (serine protease 23) that is pro-tumorigenic was significantly downregulated (10 ± 1-fold). Conclusion The dramatic upregulation of FN 1 and downregulation of ANGPTL 4 genes in PC3 cell line stably expressing U94 implicate up-regulation of FN 1 and downregulation of ANGPTL 4 in anti tumor activity of U94. Further studies are necessary to determine functional roles of differentially expressed genes in U94 recombinant PC3 cell line, and hopefully provide leads to potential therapeutic targets in prostate cancer.

The introduction of genome engineering technology has transformed biomedical research, making it possible to make precise changes to genetic information. However, creating an efficient gene-editing system requires a deep understanding of CRISPR technology, and the complex experimental systems under investigation. While Large Language Models (LLMs) have shown promise in various tasks, they often lack specific knowledge and struggle to accurately solve biological design problems. In this work, we introduce CRISPR-GPT, an LLM agent augmented with domain knowledge and external tools to automate and enhance the design process of CRISPR-based gene-editing experiments. CRISPR-GPT leverages the reasoning ability of LLMs to facilitate the process of selecting CRISPR systems, designing guide RNAs, recommending cellular delivery methods, drafting protocols, and designing validation experiments to confirm editing outcomes. We showcase the potential of CRISPR-GPT for assisting non-expert researchers with gene-editing experiments from scratch and validate the agent's effectiveness in a real-world use case. Furthermore, we explore the ethical and regulatory considerations associated with automated gene-editing design, highlighting the need for responsible and transparent use of these tools. Our work aims to bridge the gap between biological researchers across various fields with CRISPR genome engineering technology and demonstrate the potential of LLM agents in facilitating complex biological discovery tasks.

CRISPR/Cas-based gene-editing technologies have emerged as one of the most transformative tools in genome science over the past decade, providing unprecedented possibilities for both fundamental and translational research. Following the initial wave of innovations for gene knock-out, epigenetic/RNA modulation, and nickase-mediated base-editing, recent efforts have pivoted towards long-sequence gene editing- specifically, the insertion of large fragments (>1 kb) into the endogenous genome. In this review, we survey the development of these CRISPR/Cas-based sequence insertion methodologies in conjunction with the emergence of novel families of editing enzymes, such as transposases, single-stranded DNA-annealing proteins, recombinases, and integrases. Despite facing a number of challenges, this field continues to evolve rapidly and holds the potential to catalyze a new wave of revolutionary biomedical applications.

Background: Vaccination remains a potent way to curb the present covid-19 global pandemic., Objectives: To assess knowledge, perception and willingness to receive covid-19 vaccines among tertiary students in Nigeria., Methods: In the descriptive cross-sectional study, a sample size of 750 respondents was randomly selected from a university, polytechnic and college of education (COE) in Osun State, Nigeria. Independent sample T and Pearson correlation tests were used to analyse the responses., Results: There was a significant increase in the percentage score of poor perception, relative to good perception among the university and polytechnic respondents. Among the COE respondents, significant increases in the percentage scores of poor knowledge, perception and willingness to receive covid-19 vaccines, relative to the good variables were observed. Weak positive correlations between knowledge and willingness & perception and willingness to receive covid-19 vaccines among all the respondents were noted. In addition, there was a significant increase in good perception to covid-19 vaccines among university and COE, relative to polytechnic respondents. Asides, a significant increase in good willingness to receive covid-19 vaccines was observed among the university, compared to COE respondents., Conclusion: There is poor knowledge, perception and willingness to receive covid-19 vaccines among tertiary students in Osun State, Nigeria., Competing Interests: None declared., (© 2023 Johnson WA et al.)

Inpatient capacity constraints have been a pervasive challenge for hospitals throughout the COVID-19 pandemic. The Mayo Clinic Health System - Southwest Minnesota region primarily serves patients in rural southwestern Minnesota and part of Iowa and consists of 1 postacute care hospital, 1 tertiary care medical center, and 3 critical access hospitals. The main hub, Mayo Clinic Health System in Mankato, Minnesota, has a pediatric unit with dedicated pediatric hospitalists. To address the growing demand for adult inpatient beds at the height of the pandemic, the pediatric unit was opened to allow adult patients to be admitted when necessary. For several months, adult inpatient capacity exceeded 90%, which decreased the number of available pediatric (vs adult) beds throughout Minnesota, particularly in rural communities. Data for the health system showed that children were most affected because transfers to the next available hospitals for pediatric cases were 55 miles away or more. To address this gap, the hospital team successfully trialed a pediatric bed prioritization guideline that reduced pediatric transfers by 40%. This was accomplished by prioritizing the last remaining inpatient bed on the pediatric unit for pediatric patients only. This process not only reduced pediatric transfers but also increased unique patient admissions because of an average lower length of stay for pediatric patients compared with adult patients., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Inc. All rights reserved.)

Deep learning neural networks have improved performance in many cancer informatics problems, including breast cancer subtype classification. However, many networks experience underspecificationwheremultiplecombinationsofparametersachievesimilarperformance, bothin training and validation. Additionally, certain parameter combinations may perform poorly when the test distribution differs from the training distribution. Embedding prior knowledge from the literature may address this issue by boosting predictive models that provide crucial, in-depth information about a given disease. Breast cancer research provides a wealth of such knowledge, particularly in the form of subtype biomarkers and genetic signatures. In this study, we draw on past research on breast cancer subtype biomarkers, label propagation, and neural graph machines to present a novel methodology for embedding knowledge into machine learning systems. We embed prior knowledge into the loss function in the form of inter-subject distances derived from a well-known published breast cancer signature. Our results show that this methodology reduces predictor variability on state-of-the-art deep learning architectures and increases predictor consistency leading to improved interpretation. We find that pathway enrichment analysis is more consistent after embedding knowledge. This novel method applies to a broad range of existing studies and predictive models. Our method moves the traditional synthesis of predictive models from an arbitrary assignment of weights to genes toward a more biologically meaningful approach of incorporating knowledge., (Copyright © 2021 The Author(s). Published by Elsevier Ltd.. All rights reserved.)

Introduction: Orthopaedic surgery is among the least diverse fields in all of medicine. To promote the recruitment of minorities, a commonly proposed strategy is to increase the exposure of minority medical students to orthopaedic surgeons and residents who are minorities themselves. This study examines the degree to which the racial/ethnic diversity of the orthopaedic faculty and residency program influences underrepresented in medicine (URM) medical students at that institution to pursue a career in orthopaedics., Methods: Using data provided by the Association of American Medical Colleges, we identified all US medical schools that were affiliated with an orthopaedic department and an orthopaedic residency program (n = 110). For each institution, data were collected on URM representation among the orthopaedic faculty and residents (2013 to 2017), as well as the proportion of URM medical students who applied to an orthopaedic residency program (2014 to 2018). The association between institutional factors and the URM medical student orthopaedic application rate was then assessed., Results: Of 11,887 URM students who graduated from medical school during the 5-year study period, 647 applied to an orthopaedic residency program (5.4%). URM students who attended medical school at institutions with high URM representation on the orthopaedic faculty were more likely to apply in orthopaedics (odds ratio 1.27, 95% confidence interval 1.04 to 1.55, P = 0.020), as were URM students at institutions with high URM representation in the residency program (odds ratio 1.45, 95% confidence interval 1.17 to 1.79, P < 0.001)., Discussion: The benefits of a diverse orthopaedic workforce are widely acknowledged. In this study, we found that increased URM representation among the orthopaedic faculty and residents was associated with a greater likelihood that URM medical students at that institution would apply in orthopaedics. We also suggest a set of strategies to break the cycle and promote the recruitment of minorities into the field of orthopaedic surgery.

The heliobacterial reaction center (HbRC) is the simplest known photochemical reaction center, in terms of its polypeptide composition. In the heliobacterial cells, its electron donor is a cytochrome (cyt) c 553 attached to the membrane via a covalent linkage with a diacylglycerol. We have reconstituted purified HbRC into liposomes mimicking the phospholipid composition of heliobacterial membranes. We also incorporated a lipid with a headgroup containing Ni(II):nitrilotriacetate (NTA) to provide a binding site for the soluble version of the heliobacterial cyt c 553 in which the N-terminal membrane attachment site is replaced by a hexahistidine tag. The HbRC was inserted into the liposomes with the donor side preferentially exposed to the exterior; this bias increased to nearly 100% with higher concentrations (≥ 10 mol%) of the Ni(II)-NTA lipid in the membrane, and is most likely due to the net negative charge of the surface of the membrane. The HbRC in proteoliposomes without the Ni(II)-NTA lipid exhibited normal charge separation and subsequent charge recombination of the P 800 + F X - state in 15 ms; however, the oxidized primary donor (P 800 + ) was not significantly reduced by added H 6 -cyt c 553 . In contrast, with proteoliposomes containing the Ni(II)-NTA lipid, addition of H 6 -cyt c 553 resulted in a new kinetic component resulting from fast reduction (2-5 ms) of P 800 + by H 6 -cyt c 553 . The contribution of this kinetic component varied with the concentration of added H 6 -cyt c 553 and could represent 80% or more of the total P 800 + decay. Thus, the HbRC and its interaction with its native electron donor have been reconstituted into an artificial membrane system.

Highly dynamic geographical topology, two-direction mobility, and varying traffic density can lead to fairness issues in Vehicular Ad-hoc Networks (VANETs). The Medium Access Control (MAC) protocol plays a vital role in sharing the common wireless channel efficiently between vehicles in a VANET system. However, ensuring fairness between vehicles can be a challenge in designing MAC protocols for VANET systems. The existing protocol, IEEE 802.11 DCF, ensures that the packet transmission rate for a particular vehicle is directly proportional to the amount of time a vehicle spends within a service area, but it does not guarantee that faster vehicles will be able to send the minimum number of packets. Other existing MAC protocols based on IEEE 802.11 are able to provide a minimum amount of data transmission regardless of velocity, but are unable to provide an amount of data transmission that is more proportionate to the time a vehicle spends in the service area. To address the above limitations, we propose a Speed Aware Fairness Enabled MAC (SAFE-MAC) protocol that calculates the residence time of a vehicle in a service area by using mobility metrics such as position, direction, and speed to synthesize the transmission probability of each individual vehicle with respect to its residence time. This is achieved by dynamically altering the values of parameters such as minimum contention window, maximum backoff stage, and retransmission limit in the MAC protocol. We then develop an analytical model to compare the performance of our proposed protocol with contemporary MAC protocols. Numerical analysis results show that our proposed protocol significantly improves fairness among the speed-varying vehicles in VANET.

Signals from two different membrane proteins are combined to modulate how strongly sensory neurons respond to mechanical force.

Viral protein U (Vpu) is a type-III integral membrane protein encoded by Human Immunodeficiency Virus-1 (HIV- 1). It is expressed in infected host cells and plays several roles in viral progeny escape from infected cells, including down-regulation of CD4 receptors. But key structure/function questions remain regarding the mechanisms by which the Vpu protein contributes to HIV-1 pathogenesis. Here we describe expression of Vpu in bacteria, its purification and characterization. We report the successful expression of PelB-Vpu in Escherichia coli using the leader peptide pectate lyase B (PelB) from Erwinia carotovora. The protein was detergent extractable and could be isolated in a very pure form. We demonstrate that the PelB signal peptide successfully targets Vpu to the cell membranes and inserts it as a type I membrane protein. PelB-Vpu was biophysically characterized by circular dichroism and dynamic light scattering experiments and was shown to be an excellent candidate for elucidating structural models.

Root lesion nematodes (Pratylenchus spp.) hinder dryland wheat production worldwide. Montana, a leading U.S. wheat production region, has climatic conditions favorable for Pratylenchus spp. A 2006-2007 statewide soil survey revealed damaging populations of Pratylenchus neglectus, primarily in winter wheat production areas of Montana, whereas P. thornei was not found. Analyses of wheat yields in infested fields revealed negative correlations between yields and spring nematode populations (all P < 0.05 and all R 2 > 0.2). Statewide yield losses due to root lesion nematodes were an estimated 12 and 15% for winter wheat in 2006 and 2007, respectively. A subsequent study conducted in 2008 to 2009 revealed significant differences in reproductive success of P. neglectus among seven rotation treatments (P < 0.001). Nematode populations persisted from spring to fall under fallow, barley, pea, and camelina; increased under winter wheat and canola; and decreased under lentil. Populations were sustained through winter following winter wheat and barley but declined following canola, camelina, pea, lentil, and fallow. A screening of 19 barley lines for resistance to P. neglectus revealed significant variation in resistance among entries (P < 0.001), with 'Harrington' barley displaying promising levels of resistance. Development of resistant wheat cultivars remains a principal goal in managing this nematode.

Background: There is growing evidence that women diagnosed with cancer can experience intimate partner violence (IPV). This combined experience of cancer and abuse can have a profound effect on health and treatment outcomes for these cancer survivors., Objective: The purpose of this literature review was to assess the presence of IPV among female cancer survivors and to provide oncology nurses with clinical guidelines about IPV., Methods: A systematic strategy was used to locate original research from 4 databases: CINAHL, PubMed, PsycINFO, and Web of Science. Key words were used to help identify articles that focused on cancer, abuse, treatment decision making, and clinical guidelines., Results: The 10 selected articles that met the inclusion criteria were published between 2005 and 2014. The reviewed studies provided accounts of abusive partner behaviors toward women throughout their cancer trajectory. Global organizations provided the framework for clinical guidelines., Conclusions: Challenges faced by women who are concurrently living with IPV and cancer survivorship can impact treatment decision making and health outcomes. The assessment and recognition of IPV by oncology nurses are essential to help create a clinical environment in which patients feel safe and supported., Implications for Practice: This review includes clinical guidelines and describes legal considerations for oncology nurses to consider when they address and respond to IPV in their practice. The use of clinical guidelines that focus on IPV can provide standardized care in the oncology setting that can further help to meet the needs of these women.

Background: Philadelphia's Healthy Kids, Healthy Communities Out-of-School Time (OST) Initiative led by the Health Promotion Council developed 10 Healthy Living Guidelines for Out-of-School Time Programs to support a healthy environment for Philadelphia youth in 200 OST programs. Health Promotion Council participated in an enhanced cross-site evaluation with the national Robert Wood Johnson Foundation evaluator during the final year of funding to learn more about food and nutrition in the OST setting, and to share data and engage the OST food providers., Methods: A plate waste study measured the quantity and nutrition composition of meals served to youth compared to food not eaten. Staff interviews measured program adherence to the Healthy Living Guidelines, along with other facets of the food environment in 7 Philadelphia OST programs. Online surveys were sent to OST programs. Food providers (sponsors) were interviewed using Appreciative Inquiry methodology to gain insight into systems, goals, and common interests., Results: Aggregated plate waste data from 7 observations showed an average of 42% of the food uneaten after meal/snack consumption, and high sodium content of the meals. Twenty-nine staff at OST programs completed the online survey, and 3 food sponsors were interviewed. Recommendations to improve the food included increasing variety, offering more salads, fruits and vegetables, and culturally appropriate foods. Food sponsor interviews showed a variety of meal production, distribution, service and training systems, and an interest in working together. The food sponsors met to review the data and prioritized common goals. They continue to work together to improve systems and meals for food service., Conclusions: A food sponsors work group formed and continues as a result of sharing enhanced cross-site data about food in OST settings. Food sponsors continue to work together to improve systems and nutritional offerings for Philadelphia OST programs.

Background: The complex Drosophila larval peripheral nervous system, capable of monitoring sensory input from the external environment, includes a family of multiple dendritic (md) neurons with extensive dendritic arbors tiling the inner surface of the larval body wall. The class IV multiple dendritic (mdIV) neurons are the most complex with dendritic nerve endings forming direct intimate contacts with epithelial cells of the larval body wall. Functioning as polymodal mechanonociceptors with the ability to respond to both noxious mechanical stimulation and noxious heat, the mdIV neurons are also activated by nanomolar levels of the endogenous reactive oxygen species (ROS), H2O2. Although often associated with tissue damage related to oxidative stress, endogenous ROS have also been shown to function as signaling molecules at lower concentrations. The overall role of ROS in sensory signaling is poorly understood but the acutely sensitive response of mdIV neurons to ROS-mediated activation is consistent with a routine role in the regulation of mdIV neuronal activity. Larvae respond to short wavelength ultraviolet (UVC) light with an immediate and visual system-independent writhing and twisting of the body previously described as a nociceptive response. Molecular and cellular mechanisms mediating this response and potential relationships with ROS generation are not well understood. We have used the UVC-induced writhing response as a model for investigation of the proposed link between endogenous ROS production and mdIV neuron function in the larval body wall., Results: Transgenic inactivation of mdIV neurons caused a strong suppression of UVC-induced writhing behavior consistent with a key role for the mdIV neurons as mediators of the behavioral response. Direct imaging of ROS-activated fluorescence showed that UVC irradiation caused a significant increase in endogenous ROS levels in the larval body wall and transgenic overexpression of antioxidant enzymes strongly suppressed the UVC-induced writhing response. Direct electrophysiological recordings demonstrated that UVC irradiation also increased neuronal activity of the mdIV neurons., Conclusions: Results obtained using UVC irradiation to induce ROS generation provide evidence that UVC-induced writhing behavior is mediated by endogenous production of ROS capable of activating mdIV mechanonociceptors in the larval body wall.

Reactive oxygen species (ROS) in excess have been implicated in numerous chronic illnesses, including asthma, diabetes, aging, cardiovascular disease, and neurodegenerative illness. However, at lower concentrations, ROS can also serve essential routine functions as part of cellular signal transduction pathways. As products of atmospheric oxygen, ROS-mediated signals can function to coordinate external environmental conditions with growth and development. A central challenge has been a mechanistic distinction between the toxic effects of oxidative stress and endogenous ROS functions occurring at much lower concentrations. Drosophila larval aerotactic behavioral assays revealed strong developmentally regulated aversion to mild hyperoxia mediated by H2O2-dependent activation of class IV multidendritic (mdIV) sensory neurons expressing the Degenerin/epithelial Na(+) channel subunit, Pickpocket1 (PPK1). Electrophysiological recordings in foraging-stage larvae (78-84 h after egg laying [AEL]) demonstrated PPK1-dependent activation of mdIV neurons by nanomolar levels of H2O2 well below levels normally associated with oxidative stress. Acute sensitivity was reduced > 100-fold during the larval developmental transition to wandering stage (> 96 h AEL), corresponding to a loss of hyperoxia aversion behavior during the same period. Degradation of endogenous H2O2 by transgenic overexpression of catalase in larval epidermis caused a suppression of hyperoxia aversion behavior. Conversely, disruption of endogenous catalase activity using a UAS-CatRNAi transposon resulted in an enhanced hyperoxia-aversive response. These results demonstrate an essential role for low-level endogenous H2O2 as an environment-derived signal coordinating developmental behavioral transitions.

In general, Armadillidium vulgare (Latreille) are considered nonpests of soybean [Glycine max (L.) Merrill], but changes in soil conservation practices have shifted the pest status of this organism from an opportunistic to a perennial, early-season pest in parts of central Kansas. As a result, soybean producers that rotate with corn (Zea mays L.) under conservation tillage practices have resorted to removing excess corn residue by using controlled burns. In a 2-yr field study (2009-2010), we demonstrated that residue removal in burned compared with unburned plots (measured as previous crop residue weights) had minimal impact on numbers of live and dead A. vulgare, soybean seedling emergence, and isopod feeding damage over time. Specifically, removal of residue by burning did not result in higher emergence rates for soybean stands or less feeding damage by A. vulgare. In a separate study, we found that number of live A. vulgare and residue weights had no consistent relationship with seedling emergence or feeding damage. Furthermore, seedling emergence was not impacted by higher numbers ofA. vulgare in unburned plots, indicating that emergence in this study may have been influenced by factors other than A. vulgare densities. These studies demonstrate that removing residue through controlled burning did not impact seedling emergence in presence of A. vulgare and that residue and feeding damage to seedlings did not consistently relate to A. vulgare densities. Other factors that may have influenced a relationship between residue and live isopod numbers, such as variable moisture levels, are discussed.

A subset of sensory neurons embedded within the Drosophila larval body wall have been characterized as high-threshold polymodal nociceptors capable of responding to noxious heat and noxious mechanical stimulation. They are also sensitized by UV-induced tissue damage leading to both thermal hyperalgesia and allodynia very similar to that observed in vertebrate nociceptors. We show that the class IV multiple-dendritic(mdIV) nociceptors are also required for a normal larval aversion to locomotion on to a dry surface environment. Drosophila melanogaster larvae are acutely susceptible to desiccation displaying a strong aversion to locomotion on dry surfaces severely limiting the distance of movement away from a moist food source. Transgenic inactivation of mdIV nociceptor neurons resulted in larvae moving inappropriately into regions of low humidity at the top of the vial reflected as an increased overall pupation height and larval desiccation. This larval lethal desiccation phenotype was not observed in wild-type controls and was completely suppressed by growth in conditions of high humidity. Transgenic hyperactivation of mdIV nociceptors caused a reciprocal hypersensitivity to dry surfaces resulting in drastically decreased pupation height but did not induce the writhing nocifensive response previously associated with mdIV nociceptor activation by noxious heat or harsh mechanical stimuli. Larvae carrying mutations in either the Drosophila TRP channel, Painless, or the degenerin/epithelial sodium channel subunit Pickpocket1(PPK1), both expressed in mdIV nociceptors, showed the same inappropriate increased pupation height and lethal desiccation observed with mdIV nociceptor inactivation. Larval aversion to dry surfaces appears to utilize the same or overlapping sensory transduction pathways activated by noxious heat and harsh mechanical stimulation but with strikingly different sensitivities and disparate physiological responses.

In volleyball, blocking is highly correlated with team success. The identification of specific techniques that produce a more successful block would be helpful knowledge for coaches and players. This study compared the traditional, swing, and "chicken wing" blocking techniques in combination with the running step footwork pattern in order to determine which technique enabled athletes to perform a more effective block. High-speed videography (7 cameras, Vicon Motion Analysis System) was used to capture the blocking movements of thirteen female NCAA Division I athletes (age = 19.4 ± 1.19 years, height = 1.82 ± 0.08 m, mass = 70.63 ± 7.96 kg, and years of participation at the collegiate level = 2.23 ± 1.17 years). Each player was familiar with each blocking technique. Reflective markers were placed on the players and in randomized order the players performed 3 blocking trials of each technique. The following dependent variables were assessed: The time it took the athletes to get off the ground and get their hands above (vertically) the net was calculated. The distance the hand reached over the net or hand penetration (displacement between the net and finger in the anterior and vertical planes) was also measured. Lastly, jump height was calculated. Repeated measures ANOVA and post-hoc comparisons were done (α = 0.05). There was no significant difference in the main effect for time to get off the ground (p > 0.05). There was a significant difference in the time to get the hands above the net (p < 0.05). The swing block was best for jump height (p <.001) and hand penetration (p < 0.05). These results can help coaches and players decide which blocking technique will benefit them most as a blocking team and as individual blockers. Key pointsThe swing blocking technique resulted in greater jump heights and increased hand penetration, relative to the traditional and chicken wing blocking techniques.The chicken wing blocking technique resulted in greater jump heights and increased hand penetration, relative to the traditional blocking technique.THE TRADITIONAL BLOCKING TECHNIQUE DOES NOT APPEAR TO PROVIDE ANY COMPETITIVE ADVANTAGE RELATED TO THE VARIABLES OBSERVED DURING THIS STUDY: (1) duration spent getting off of the ground and placing hands over the net, (2) jump height, and (3) hand penetration magnitude.

Controlled organismal growth to an appropriate adult size requires a regulated balance between nutrient resources, feeding behavior and growth rate. Defects can result in decreased survival and/or reproductive capability. Since Drosophila adults do not grow larger after eclosion, timing of feeding cessation during the third and final larval instar is critical to final size. We demonstrate that larval food exit is preceded by a period of increased larval surfacing behavior termed the Intermediate Surfacing Transition (IST) that correlates with the end of larval feeding. This behavioral transition occurred during the larval Terminal Growth Period (TGP), a period of constant feeding and exponential growth of the animal. IST behavior was dependent upon function of a subset of peripheral sensory neurons expressing the Degenerin/Epithelial sodium channel (DEG/ENaC) subunit, Pickpocket1(PPK1). PPK1 neuron inactivation or loss of PPK1 function caused an absence of IST behavior. Transgenic PPK1 neuron hyperactivation caused premature IST behavior with no significant change in timing of larval food exit resulting in decreased final adult size. These results suggest a peripheral sensory mechanism functioning to alter the relationship between the animal and its environment thereby contributing to the length of the larval TGP and determination of final adult size., ((c) 2010 Elsevier Inc. All rights reserved.)

The three Drosophila atypical soluble guanylyl cyclases, Gyc-89Da, Gyc-89Db, and Gyc-88E, have been proposed to act as oxygen detectors mediating behavioral responses to hypoxia. Drosophila larvae mutant in any of these subunits were defective in their hypoxia escape response-a rapid cessation of feeding and withdrawal from their food. This response required cGMP and the cyclic nucleotide-gated ion channel, cng, but did not appear to be dependent on either of the cGMP-dependent protein kinases, dg1 and dg2. Specific activation of the Gyc-89Da neurons using channel rhodopsin showed that activation of these neurons was sufficient to trigger the escape behavior. The hypoxia escape response was restored by reintroducing either Gyc-89Da or Gyc-89Db into either Gyc-89Da or Gyc-89Db neurons in either mutation. This suggests that neurons that co-express both Gyc-89Da and Gyc-89Db subunits are primarily responsible for activating this behavior. These include sensory neurons that innervate the terminal sensory cones. Although the roles of Gyc-89Da and Gyc-89Db in the hypoxia escape behavior appeared to be identical, we also showed that changes in larval crawling behavior in response to either hypoxia or hyperoxia differed in their requirements for these two atypical sGCs, with responses to 15% oxygen requiring Gyc-89Da and responses to 19 and 25% requiring Gyc-89Db. For this behavior, the identity of the neurons appeared to be critical in determining the ability to respond appropriately.

Growth of multicellular organisms proceeds through a series of precisely timed developmental events requiring coordination between gene expression, behavioral changes, and environmental conditions. In Drosophila melanogaster larvae, the essential midthird instar transition from foraging (feeding) to wandering (non-feeding) behavior occurs prior to pupariation and metamorphosis. The timing of this key transition is coordinated with larval growth and size, but physiological mechanisms regulating this process are poorly understood. Results presented here show that Drosophila larvae associate specific environmental conditions, such as temperature, with food in order to enact appropriate foraging strategies. The transition from foraging to wandering behavior is associated with a striking reversal in the behavioral responses to food-associated stimuli that begins early in the third instar, well before food exit. Genetic manipulations disrupting expression of the Degenerin/Epithelial Sodium Channel subunit, Pickpocket1(PPK1) or function of PPK1 peripheral sensory neurons caused defects in the timing of these behavioral transitions. Transient inactivation experiments demonstrated that sensory input from PPK1 neurons is required during a critical period early in the third instar to influence this developmental transition. Results demonstrate a key role for the PPK1 sensory neurons in regulation of important behavioral transitions associated with developmental progression of larvae from foraging to wandering stage.

Coordination of rhythmic locomotion depends upon a precisely balanced interplay between central and peripheral control mechanisms. Although poorly understood, peripheral proprioceptive mechanosensory input is thought to provide information about body position for moment-to-moment modifications of central mechanisms mediating rhythmic motor output. Pickpocket1 (PPK1) is a Drosophila subunit of the epithelial sodium channel (ENaC) family displaying limited expression in multiple dendritic (md) sensory neurons tiling the larval body wall and a small number of bipolar neurons in the upper brain. ppk1 null mutant larvae had normal external touch sensation and md neuron morphology but displayed striking alterations in crawling behavior. Loss of PPK1 function caused an increase in crawling speed and an unusual straight path with decreased stops and turns relative to wild-type. This enhanced locomotion resulted from sustained peristaltic contraction wave cycling at higher frequency with a significant decrease in pause period between contraction cycles. The mutant phenotype was rescued by a wild-type PPK1 transgene and duplicated by expressing a ppk1RNAi transgene or a dominant-negative PPK1 isoform. These results demonstrate that the PPK1 channel plays an essential role in controlling rhythmic locomotion and provide a powerful genetic model system for further analysis of central and peripheral control mechanisms and their role in movement disorders.

The ability to detect salt is critical for the survival of terrestrial animals. Based on amiloride-dependent inhibition, the receptors that detect salt have been postulated to be DEG/ENaC channels. We found the Drosophila DEG/ENaC genes Pickpocket11 (ppk11) and Pickpocket19 (ppk19) expressed in the larval taste-sensing terminal organ and in adults on the taste bristles of the labelum, the legs, and the wing margins. When we disrupted PPK11 or PPK19 function, larvae lost their ability to discriminate low concentrations of Na(+) or K(+) from water, and the electrophysiologic responses to low salt concentrations were attenuated. In both larvae and adults, disrupting PPK11 or PPK19 affected the behavioral response to high salt concentrations. In contrast, the response of larvae to sucrose, pH 3, and several odors remained intact. These results indicate that the DEG/ENaC channels PPK11 and PPK19 play a key role in detecting Na(+) and K(+) salts.

This article explores late-onset crack use among midlife and older adults as an emerging risk factor for HIV infection. Most research on illicit drug use and HIV infection/AIDS has focused on younger drug users, typically those who inject. The initiation into crack use during later adulthood challenges this narrow view. The analysis the authors present was drawn from qualitative and quantitative data collected as part of their ongoing research on illicit drug use. The subsample consisted of 27 men who began using crack at the age of 50 or older and 40 women who started using crack at the age of 35 or older. The findings suggest a typology of late-onset users with differing forms of HIV risk and prevention needs. The authors end the article with recommendations for effective HIV risk reduction programs for late-onset crack users.

Although the ability to sense temperature is critical for many organisms, the underlying mechanisms are poorly understood. Using the calcium reporter yellow cameleon 2.1 and electrophysiological recordings, we identified thermosensitive neurons and examined their physiologic response in Drosophila melanogaster larvae. In the head, terminal sensory organ neurons showed increased activity in response to cooling by < or =1 degrees C, heating reduced their basal activity, and different units showed distinct response patterns. Neither cooling nor heating affected dorsal organ neurons. Body wall neurons showed a variety of distinct response patterns to both heating and cooling; the diverse thermal responses were strikingly similar to those described in mammals. These data establish a functional map of thermoresponsive neurons in Drosophila larvae and provide a foundation for understanding mechanisms of thermoreception in both insects and mammals.

The Drosophila tracheal system and mammalian airways are branching networks of tubular epithelia that deliver oxygen to the organism. In mammals, the epithelial Na(+) channel (ENaC) helps clear liquid from airways at the time of birth and removes liquid from the airspaces in adults. We tested the hypothesis that related Drosophila degenerin (DEG)/ENaC family members might play a similar role in the fly. Among 16 Drosophila DEG/ENaC genes, called pickpocket (PPK) genes, we found 9 expressed in the tracheal system. By in situ hybridization, expression appeared in late-stage embryos after tracheal tube formation, with individual PPK genes showing distinct temporal and spatial expression patterns as development progressed. Promoters for several PPK genes drove reporter gene expression in the larval and adult tracheal systems. Adding the DEG/ENaC channel blocker amiloride to the medium inhibited liquid clearance from the trachea of first instar larvae. Moreover, when RNA interference was used to silence PPK4 and PPK11, larvae failed to clear tracheal liquid. These data suggest substantial molecular diversity of DEG/ENaC channel expression in the Drosophila tracheal system where the PPK proteins likely play a role in Na(+) absorption. Extensive similarities between Drosophila and mammalian airways offer opportunities for genetic studies that may decipher further the structure and function of DEG/ENaC proteins and development of the airways.

Axonal selection of synaptic partners is generally believed to determine wiring specificity in the nervous system. However, we have recently found evidence for specific dendritic targeting in the olfactory system of Drosophila: second order olfactory neurons (Projection Neurons) from the anterodorsal (adPN) and lateral (lPN) lineages send their dendrites to stereotypical, intercalating but non-overlapping glomeruli. Here we show that POU domain transcription factors, Acj6 and Drifter, are expressed in adPNs and lPNs respectively, and are required for their dendritic targeting. Moreover, misexpression of Acj6 in lPNs, or Drifter in adPNs, results in dendritic targeting to glomeruli normally reserved for the other PN lineage. Thus, Acj6 and Drifter translate PN lineage information into distinct dendritic targeting specificity. Acj6 also controls stereotypical axon terminal arborization of PNs in a central target, suggesting that the connectivity of PN axons and dendrites in different brain centers is coordinately regulated.

Use of the central one third bone-patellar tendon-bone autograft is an accepted technique for anterior cruciate ligament (ACL) reconstruction. Patellar tendon rupture following ACL reconstruction is an acknowledged, although rarely reported, complication of this procedure. Of the limited patellar tendon rupture cases reported in the literature, most are described early in the postoperative period. We present a case of late patellar tendon rupture more than 3 years after uneventful ACL reconstruction in a 32-year-old man.

Objectives: To assess HIV prevalence, incidence, and associated risk factors among IDUs in Chicago., Methods: Seven hundred ninety-four street-recruited IDUs ranging in age from 18 to 50 years, who were not in drug treatment at study enrollment, were interviewed and tested for HIV at baseline and at two follow-ups scheduled 6 and 12 months after baseline. Questionnaires assessed respondents' demographic characteristics, medical and drug treatment histories, drug use, and sexual practices., Results: HIV seroprevalence at baseline was 18%. Logistic regression identified the following determinants of prevalent HIV infection: Puerto Rican ethnicity, homosexual or bisexual self-identification, injecting for 4 or more years, and having smoked crack cocaine in the past 6 months. Follow-up data were collected from 584 (73.6%) participants. Mean duration of follow-up was 16.5 months, indicating that most subjects had follow-up intervals longer than the scheduled 6 and 12 months. Seven HIV seroconversions were observed in 632 person years of risk, yielding an incidence rate of 1.1 per 100 person years of risk. Injection for 3 or less years was positively associated with HIV seroconversion., Conclusions: The findings provide evidence of a decline in HIV incidence among IDUs, though newer injectors remain at elevated risk for infection.

The Drosophila Vestigial protein has been shown to play an essential role in the regulation of cell proliferation and differentiation within the developing wing imaginal disc. Cell-specific expression of vg is controlled by two separate transcriptional enhancers. The boundary enhancer controls expression in cells near the dorsoventral (DV) boundary and is regulated by the Notch signal transduction pathway, while the quadrant enhancer responds to the Decapentaplegic and Wingless morphogen gradients emanating from cells near the anteroposterior (AP) and DV boundaries, respectively. MAD-dependent activation of the vestigial quadrant enhancer results in broad expression throughout the wing pouch but is excluded from cells near the DV boundary. This has previously been thought to be due to direct repression by a signal from the DV boundary; however, we show that this exclusion of quadrant enhancer-dependent expression from the DV boundary is due to the absence of an additional essential activator in those cells. The Drosophila POU domain transcriptional regulator, Drifter, is expressed in all cells within the wing pouch expressing a vgQ-lacZ transgene and is also excluded from the DV boundary. Viable drifter hypomorphic mutations cause defects in cell proliferation and wing vein patterning correlated with decreased quadrant enhancer-dependent expression. Drifter misexpression at the DV boundary using the GAL4/UAS system causes ectopic outgrowths at the distal wing tip due to induction of aberrant Vestigial expression, while a dominant-negative Drifter isoform represses expression of vgQ-lacZ and causes severe notching of the adult wing. In addition, we have identified an essential evolutionarily conserved sequence element bound by the Drifter protein with high affinity and located adjacent to the MAD binding site within the quadrant enhancer. Our results demonstrate that Drifter functions along with MAD as a direct activator of Vestigial expression in the wing pouch.

During Drosophila embryogenesis the CNS midline cells have organizing activities that are required for proper elaboration of the axon scaffold and differentiation of neighboring neuroectodermal and mesodermal cells. CNS midline development is dependent on Single-minded (Sim), a basic-helix-loop-helix (bHLH)-PAS transcription factor. We show here that Fish-hook (Fish), a Sox HMG domain protein, and Drifter (Dfr), a POU domain protein, act in concert with Single-minded to control midline gene expression. single-minded, fish-hook, and drifter are all expressed in developing midline cells, and both loss- and gain-of-function assays revealed genetic interactions between these genes. The corresponding proteins bind to DNA sites present in a 1 kb midline enhancer from the slit gene and regulate the activity of this enhancer in cultured Drosophila Schneider line 2 cells. Fish-hook directly associates with the PAS domain of Single-minded and the POU domain of Drifter; the three proteins can together form a ternary complex in yeast. In addition, Fish can form homodimers and also associates with other bHLH-PAS and POU proteins. These results indicate that midline gene regulation involves the coordinate functions of three distinct types of transcription factors. Functional interactions between members of these protein families may be important for numerous developmental and physiological processes.