High frequency and mortality of head and neck malignancy in organ transplant recipients in Ireland - A national cohort study.

BACKGROUND: Solid organ transplant recipients are recognized to carry a high burden of malignancy and frequently this cancer develops in the head and neck region. Furthermore, cancer of the head and neck post-transplant carries a significantly increased mortality. In this study, we aim to conduct a national retrospective cohort study to investigate the impact of head and neck cancer in terms of frequency and mortality in a large group of solid organ transplant recipients over a 20 year time span and compare the mortality in transplant patients to non-transplant patients with head and neck cancer. METHODS: Patients in the Republic of Ireland who underwent solid organ transplantation between 1994 and 2014 who developed post-transplant head and neck malignancy were identified from the records of two prospective, national databases (National Cancer Registry of Ireland (NCRI) and The Irish Transplant Cancer Group database) working in conjunction with each other. Incidence of head and neck malignancy post-transplant was compared with the general population by means of standardised incidence ratios (SIR). Cumulative incidence of all cause and cancer related mortality from head and neck keratinocytic was undertaken by a competing risks analysis. RESULTS: A total of 3346 solid organ transplant recipients were identified, 2382 (71.2 %) kidney, 562 (16.8 %) liver, 214 (6.4 %) cardiac and 188 (5.6 %) lung. During the period of follow up of 428 patients developed head and neck cancer, representing (12.8 %) of the population. 97 % of these patients developed keratinocytic cancers, specifically, of head and neck. The frequency of post-transplant head and neck cancer was related to the duration of immunosuppression with 14 % of patients developing cancer at 10 years and 20 % having developed at least one cancer by 15 years. 12 (3 %) patients developed non-cutaneous head and neck malignancy. 10 (0.3 %) patients died due to head and neck keratinocytic malignancy post-transplant. Competing risk analysis demonstrated that organ transplantation conferred a strong independent effect of death, compared to non-transplant patients with head and neck keratinocytes. This applied specifically for kidney (HR 4.4, 95 % CI 2.5-7.8) and heart transplants (HR 6.5, 95 % CI 2.1-19.9), and overall, across the four transplant categories (P < 0.001). The SIR of developing keratinocyte cancer varied based on primary tumor site, gender, and type of transplant organ. CONCLUSION: Transplant patients demonstrate a particularly high rate of head and neck keratinocyte cancer with a very high rate of associated mortality. Physicians should be cognizant of the increased rate of malignancy in this population and monitor for red flag signs/symptoms.

Impact of Artificial Intelligence Assistance on Chest CT Interpretation Times: A Prospective Randomized Study.

BACKGROUND. Deep learning-based convolutional neural networks have enabled major advances in development of artificial intelligence (AI) software applications. Modern AI applications offer comprehensive multiorgan evaluation. OBJECTIVE. The purpose of this article was to evaluate the impact of an automated AI platform integrated into clinical workflow for chest CT interpretation on radiologists' interpretation times when evaluated in a real-world clinical setting. METHODS. In this prospective single-center study, a commercial AI software solution was integrated into clinical workflow for chest CT interpretation. The software provided automated analysis of cardiac, pulmonary, and musculoskeletal findings, including labeling, segmenting, and measuring normal structures as well as detecting, labeling, and measuring abnormalities. AI-annotated images and autogenerated summary results were stored in the PACS and available to interpreting radiologists. A total of 390 patients (204 women, 186 men; mean age, 62.8 ± 13.3 [SD] years) who underwent out-patient chest CT between January 19, 2021, and January 28, 2021, were included. Scans were randomized using 1:1 allocation between AI-assisted and non-AI-assisted arms and were clinically interpreted by one of three cardiothoracic radiologists (65 scans per arm per radiologist; total of 195 scans per arm) who recorded interpretation times using a stopwatch. Findings were categorized according to review of report impressions. Interpretation times were compared between arms. RESULTS. Mean interpretation times were significantly shorter in the AI-assisted than in the non-AI-assisted arm for all three readers (289 ± 89 vs 344 ± 129 seconds, p < .001; 449 ± 110 vs 649 ± 82 seconds, p < .001; 281 ± 114 vs 348 ± 93 seconds, p = .01) and for readers combined (328 ± 122 vs 421 ± 175 seconds, p < .001). For readers combined, the mean difference was 93 seconds (95% CI, 63-123 seconds), corresponding with a 22.1% reduction in the AI-assisted arm. Mean interpretation time was also shorter in the AI-assisted arm compared with the non-AI-assisted arm for contrast-enhanced scans (83 seconds), noncontrast scans (104 seconds), negative scans (84 seconds), positive scans without significant new findings (117 seconds), and positive scans with significant new findings (92 seconds). CONCLUSION. Cardiothoracic radiologists exhibited a 22.1% reduction in chest CT interpretations times when they had access to results from an automated AI support platform during real-world clinical practice. CLINICAL IMPACT. Integration of the AI support platform into clinical workflow improved radiologist efficiency.

Post-transplant malignancy in solid organ transplant recipients in Ireland, The Irish Transplant Cancer Group.

OBJECTIVE: Solid organ transplant recipients are at increased risk of cancer compared to the general population. To date, this risk in Ireland has not been investigated. We conducted a national registry study of cancer incidence following solid organ transplantation. METHODS: National centers for solid organ transplantation supplied their respective registry databases to cross-reference with episodes of malignancy from the National Cancer Registry Ireland (NCRI) between 1994 and 2014. Standardized incidence of cancer post-transplant was compared to the general population by means of standardized incidence ratios (SIRs), and between solid organ transplant types by incidence rate ratios. RESULTS: A total of 3346 solid organ transplant recipients were included in this study. Kidney transplant recipients constituted the majority of participants (71.2%), followed by liver (16.8%), heart (6.4%), and lung (5.6%) transplants. The most common cancers within the composite of all transplant recipients included the following (SIR [95% CI]): squamous and basal cell carcinoma (20.05 [17.97, 22.31] and 7.16 [6.43, 7.96], respectively), non-Hodgkin lymphoma (6.23 [4.26, 8.59]), and renal cell carcinoma (3.36 [1.96, 5.38]). CONCLUSIONS: This study reports the incidence of cancer following solid organ transplantation in Ireland. These results have significant national policy implications for surveillance, and early diagnosis in this patient group.

Flightin maintains myofilament lattice organization required for optimal flight power and courtship song quality in Drosophila.

The indirect flight muscles (IFMs) of Drosophila and other insects with asynchronous flight muscles are characterized by a crystalline myofilament lattice structure. The high-order lattice regularity is considered an adaptation for enhanced power output, but supporting evidence for this claim is lacking. We show that IFMs from transgenic flies expressing flightin with a deletion of its poorly conserved N-terminal domain (flnΔN62 ) have reduced inter-thick filament spacing and a less regular lattice. This resulted in a decrease in flight ability by 33% and in skinned fibre oscillatory power output by 57%, but had no effect on wingbeat frequency or frequency of maximum power output, suggesting that the underlying actomyosin kinetics is not affected and that the flight impairment arises from deficits in force transmission. Moreover, we show that flnΔN62 males produced an abnormal courtship song characterized by a higher sine song frequency and a pulse song with longer pulses and longer inter-pulse intervals (IPIs), the latter implicated in male reproductive success. When presented with a choice, wild-type females chose control males over mutant males in 92% of the competition events. These results demonstrate that flightin N-terminal domain is required for optimal myofilament lattice regularity and IFM activity, enabling powered flight and courtship song production. As the courtship song is subject to female choice, we propose that the low amino acid sequence conservation of the N-terminal domain reflects its role in fine-tuning species-specific courtship songs.

Diffusion coefficients of endogenous cytosolic proteins from rabbit skinned muscle fibers.

Efflux time courses of endogenous cytosolic proteins were obtained from rabbit psoas muscle fibers skinned in oil and transferred to physiological salt solution. Proteins were separated by gel electrophoresis and compared to load-matched standards for quantitative analysis. A radial diffusion model incorporating the dissociation and dissipation of supramolecular complexes accounts for an initial lag and subsequent efflux of glycolytic and glycogenolytic enzymes. The model includes terms representing protein crowding, myofilament lattice hindrance, and binding to the cytomatrix. Optimization algorithms returned estimates of the apparent diffusion coefficients, D(r,t), that were very low at the onset of diffusion (∼10(-10) cm(2) s(-1)) but increased with time as cytosolic protein density, which was initially high, decreased. D(r,t) at later times ranged from 2.11 × 10(-7) cm(2) s(-1) (parvalbumin) to 0.20 × 10(-7) cm(2) s(-1) (phosphofructose kinase), values that are 3.6- to 12.3-fold lower than those predicted in bulk water. The low initial values are consistent with the presence of complexes in situ; the higher later values are consistent with molecular sieving and transient binding of dissociated proteins. Channeling of metabolic intermediates via enzyme complexes may enhance production of adenosine triphosphate at rates beyond that possible with randomly and/or sparsely distributed enzymes, thereby matching supply with demand.

An evolutionary analysis of flightin reveals a conserved motif unique and widespread in Pancrustacea.

Flightin is a thick filament protein that in Drosophila melanogaster is uniquely expressed in the asynchronous, indirect flight muscles (IFM). Flightin is required for the structure and function of the IFM and is indispensable for flight in Drosophila. Given the importance of flight acquisition in the evolutionary history of insects, here we study the phylogeny and distribution of flightin. Flightin was identified in 69 species of hexapods in classes Collembola (springtails), Protura, Diplura, and insect orders Thysanura (silverfish), Dictyoptera (roaches), Orthoptera (grasshoppers), Pthiraptera (lice), Hemiptera (true bugs), Coleoptera (beetles), Neuroptera (green lacewing), Hymenoptera (bees, ants, and wasps), Lepidoptera (moths), and Diptera (flies and mosquitoes). Flightin was also found in 14 species of crustaceans in orders Anostraca (water flea), Cladocera (brine shrimp), Isopoda (pill bugs), Amphipoda (scuds, sideswimmers), and Decapoda (lobsters, crabs, and shrimps). Flightin was not identified in representatives of chelicerates, myriapods, or any species outside Pancrustacea (Tetraconata, sensu Dohle). Alignment of amino acid sequences revealed a conserved region of 52 amino acids, referred herein as WYR, that is bound by strictly conserved tryptophan (W) and arginine (R) and an intervening sequence with a high content of tyrosines (Y). This motif has no homologs in GenBank or PROSITE and is unique to flightin and paraflightin, a putative flightin paralog identified in decapods. A third motif of unclear affinities to pancrustacean WYR was observed in chelicerates. Phylogenetic analysis of amino acid sequences of the conserved motif suggests that paraflightin originated before the divergence of amphipods, isopods, and decapods. We conclude that flightin originated de novo in the ancestor of Pancrustacea > 500 MYA, well before the divergence of insects (~400 MYA) and the origin of flight (~325 MYA), and that its IFM-specific function in Drosophila is a more recent adaptation. Furthermore, we propose that WYR represents a novel myosin coiled-coil binding motif.

Neurovascular Imaging with Ultra-High-Resolution Photon-Counting CT: Preliminary Findings on Image Quality Evaluation.

BACKGROUND AND PURPOSE: The first-generation photon-counting detector (PCD) CT was recently introduced into clinical practice and represents a promising innovation in high-resolution CT imaging. The purpose of this study was to assess the image quality of ultra-high-resolution (UHR) PCD-CT compared with energy-integrating detector (EID)-CT, and to explore different reconstruction kernel sharpness levels for the evaluation of intracranial aneurysms. MATERIALS AND METHODS: Ten patients with intracranial saccular aneurysms, who had previously undergone conventional EID-CT, were prospectively enrolled. CT angiograms were acquired on a clinical dual-source PCD-CT in UHR mode, and reconstructed with four vascular kernels (Bv36, Bv40, Bv44, Bv48). Quantitative and qualitative image quality parameters of the intracranial arteries were evaluated. For the quantitative analysis (image noise, SNR, CNR), regions of interest were manually placed at standard anatomical intracranial and extracranial locations by one author. In addition, vessel border sharpness was evaluated quantitatively. For the qualitative analysis, three blinded neuroradiologists rated PCD-CT and EID-CT image quality for the evaluation of the intracranial vessels (i.e., the aneurysms and nine standard vascular branching locations) on a 5-point Likert-type scale. Additionally, readers independently selected their preferred kernel among the four kernels evaluated on PCD-CT. RESULTS: In terms of quantitative image quality, Bv48, the sharpest kernel, yielded increased image noise, and decreased SNR and CNR parameters compared to Bv36, the smoothest kernel. Compared to EID-CT, the Bv48 kernel offered better quantitative image quality for the evaluation of small intracranial vessels (p < .001). Image quality ratings of the Bv48 were superior to those of the EIDCT, and not significantly different from ratings of the B44 reconstruction kernel. When comparing side-by-side all four PCD-CT reconstruction kernels, readers selected the B48 kernel as the best to visualize the aneurysms in 80% of cases. CONCLUSIONS: UHR PCD-CT provides improved image quality for neurovascular imaging. Although the less sharp kernels provided superior SNR and CNR, the sharpest kernels delivered the best subjective image quality on PCD-CT for the evaluation of intracranial aneurysms.CNR ＝ Contrast-to-Noise Ratio; EID-CT ＝ Energy-Integrating Detector CT; PCD-CT ＝ Photon-Counting Detector CT; QIR ＝ Quantum Iterative Reconstruction; UHR ＝ Ultra-High-Resolution.

Family-based associations in measures of psychological distress and quality of life in a cardiac screening clinic for inheritable cardiac diseases: a cross-sectional study.

BACKGROUND: Family-based cardiac screening programmes for persons at risk for genetic cardiac diseases are now recommended. However, the psychological wellbeing and health related quality of life (QoL) of such screened patients is poorly understood, especially in younger patients. We sought to examine wellbeing and QoL in a representative group of adults aged 16 and over in a dedicated family cardiac screening clinic. METHODS: Prospective survey of consecutive consenting patients attending a cardiac screening clinic, over a 12 month period. Data were collected using two health measurement tools: the Short Form 12 (version 2) and the Hospital Anxiety and Depression Scale (HADS), along with baseline demographic and screening visit-related data. The HADS and SF-12v.2 outcomes were compared by age group. Associations with a higher HADS score were examined using logistic regression, with multi-level modelling used to account for the family-based structure of the data. RESULTS: There was a study response rate of 86.6%, with n=334 patients providing valid HADS data (valid response rate 79.5%), and data on n=316 retained for analysis. One-fifth of patients were aged under 25 (n=61). Younger patients were less likely than older to describe significant depression on their HADS scale (p<0.0001), although there were overall no difference between the prevalence of a significant HADS score between the younger and older age groups (18.0% vs 20.0%, p=0.73). Significant positive associates of a higher HADS score were having lower educational attainment, being single or separated, and being closely related to the family proband. Between-family variance in anxiety and depression scores was greater than within-family variance. CONCLUSIONS: High levels of anxiety were seen amongst patients attending a family-based cardiac screening clinic.Younger patients also had high rates of clinically significant anxiety. Higher levels of anxiety and depression tends to run in families, and this has implications for family screening and intervention programmes.

Courtship song analysis of Drosophila muscle mutants.

As part of the mating ritual, males of Drosophila species produce species-specific courtship songs through wing vibrations generated by the thoracic musculature. While previous studies have shown that indirect flight muscles (IFM) are neurally activated during courtship song production, the precise role of these muscles in song production has not been investigated. Fortunately, IFM mutants abound in Drosophila melanogaster and studies spanning several decades have shed light on the role of muscle proteins in IFM-powered flight. Analysis of courtship songs in these mutants offers the opportunity to uncover the role of the IFM in a behavior distinct than flight and subject to different evolutionary selection regimes. Here, we describe protocols for the recording and analysis of courtship behavior and mating song of D. melanogaster muscle transgenic and mutant strains. To record faint acoustic signal of courtship songs, an insulated mating compartment was used inside a recording device (INSECTAVOX) equipped with a modified electret microphone, a low-noise power supply, and noise filters. Songs recorded in the INSECTAVOX are digitized using Goldwave, whose several features enable extraction of critical song parameters, including carrier frequencies for pulse song and sine song. We demonstrate the utility of this approach by showing that deletion of the N-terminal region of the myosin regulatory light chain, a mutation known to decrease wing beat frequency and flight power, affects courtship song parameters.

Neutron imaging of ion transport in mesoporous carbon materials.

Neutron imaging is presented as a tool for quantifying the diffusion of ions inside porous materials, such as carbon electrodes used in the desalination process via capacitive deionization and in electrochemical energy-storage devices. Monolithic mesoporous carbon electrodes of ∼10 nm pore size were synthesized based on a soft-template method. The electrodes were used with an aqueous solution of gadolinium nitrate in an electrochemical flow-through cell designed for neutron imaging studies. Sequences of neutron images were obtained under various conditions of applied potential between the electrodes. The images revealed information on the direction and magnitude of ion transport within the electrodes. From the time-dependent concentration profiles inside the electrodes, the average value of the effective diffusion coefficient for gadolinium ions was estimated to be 2.09 ± 0.17 × 10(-11) m(2) s(-1) at 0 V and 1.42 ± 0.06 × 10(-10) m(2) s(-1) at 1.2 V. The values of the effective diffusion coefficient obtained from neutron imaging experiments can be used to evaluate model predictions of the ion transport rate in capacitive deionization and electrochemical energy-storage devices.

BAITING AND BANDING: EXPERT OPINION ON HOW BAIT TRAPPING MAY INFLUENCE THE OCCURRENCE OF HIGHLY PATHOGENIC AVIAN INFLUENZA (HPAI) AMONG DABBLING DUCKS.

A Eurasian lineage highly pathogenic avian influenza virus (HPAIV) of the clade 2.3.4.4b (Goose/Guangdong lineage) was detected in migratory bird populations in North America in December 2021, and it, along with its reassortants, have since caused wild and domestic bird outbreaks across the continent. Relative to previous outbreaks, HPAIV cases among wild birds in 2022 exhibited wider geographic extent within North America and higher levels of mortality, suggesting the potential for population-level impacts. Given the possible conservation implications of HPAIV in wild birds, natural resource managers have sought guidance on actions that may mitigate negative effects of disease among North American bird populations, including modification of existing management practices. Banding of waterfowl is a critical tool for population management for several harvested species in North America, but some banding techniques, such as bait trapping, can lead to increased congregation of waterfowl, potentially altering HPAIV transmission. We used an expert opinion exercise to assess how bait trapping of dabbling ducks in Canada may influence HPAIV transmission and wild bird health. The expert group found that it is moderately likely that bait trapping of dabbling ducks in wetlands will significantly increase the transmission of HPAIV among individual ducks, but there is a low probability that this will result in significant population-level effects on North American dabbling ducks. Considering the lack of empirical work studying how capture and handling methods may change transmission of HPAIV among waterfowl, as well as the importance of bait trapping for waterfowl management in North America, future work should focus on filling knowledge gaps pertaining to the influence of baiting on HPAIV occurrence to better inform banding procedures and management decision making.

AI-based, automated chamber volumetry from gated, non-contrast CT.

BACKGROUND: Accurate chamber volumetry from gated, non-contrast cardiac CT (NCCT) scans can be useful for potential screening of heart failure. OBJECTIVES: To validate a new, fully automated, AI-based method for cardiac volume and myocardial mass quantification from NCCT scans compared to contrasted CT Angiography (CCTA). METHODS: Of a retrospectively collected cohort of 1051 consecutive patients, 420 patients had both NCCT and CCTA scans at mid-diastolic phase, excluding patients with cardiac devices. Ground truth values were obtained from the CCTA scans. RESULTS: The NCCT volume computation shows good agreement with ground truth values. Volume differences [95% CI ] and correlation coefficients were: -9.6 [-45; 26] mL, r â€‹= â€‹0.98 for LV Total, -5.4 [-24; 13] mL, r â€‹= â€‹0.95 for LA, -8.7 [-45; 28] mL, r â€‹= â€‹0.94 for RV, -5.2 [-27; 17] mL, r â€‹= â€‹0.92 for RA, -3.2 [-42; 36] mL, r â€‹= â€‹0.91 for LV blood pool, and -6.7 [-39; 26] g, r â€‹= â€‹0.94 for LV wall mass, respectively. Mean relative volume errors of less than 7% were obtained for all chambers. CONCLUSIONS: Fully automated assessment of chamber volumes from NCCT scans is feasible and correlates well with volumes obtained from contrast study.

Self-reported beta-lactam allergy in government and private hospitals in Cape Town, South Africa.

BACKGROUND: Up to a quarter of inpatients in high-income countries (HICs) self-report beta-lactam allergy (BLA), which if incorrect,increases the use of alternative antibiotics, worsening individual health outcomes and driving bacterial resistance. In HICs, up to 95% ofself-reported BLAs are incorrect. The epidemiology of BLA in low- and middle-income African countries is unknown. OBJECTIVES: To describe the epidemiology and de-labelling outcomes of self-reported BLA in hospitalised South African (SA) patients. METHODS: Point-prevalence surveys were conducted at seven hospitals (adult, paediatric, government and privately funded, district andtertiary level) in Cape Town, SA, between April 2019 and June 2021. Ward prescription records and in-person interviews were conductedto identify and risk-stratify BLA patients using the validated PEN-FAST tool. De-labelling was attempted at the tertiary allergy clinic atGroote Schuur Hospital. RESULTS: A total of 1 486 hospital inpatients were surveyed (1 166 adults and 320 children). Only 48 patients (3.2%) self-reported a BLA,with a higher rate in private than in government-funded hospitals (6.3% v. 2.8%; p=0.014). Using the PEN-FAST tool, only 10.4% (n=5/48)of self-reported BLA patients were classified as high risk for true penicillin hypersensitivity. Antibiotics were prescribed to 70.8% (n=34/48)of self-reported BLA patients, with 64.7% (n=22/34) receiving a beta-lactam. Despite three attempts to contact patients for de-labelling atthe allergy clinic, only 3/36 underwent in vivo testing, with no positive results, and 1 patient proceeded to a negative oral challenge. CONCLUSION: Unlike HICs, self-reported BLA is low among inpatients in SA. The majority of those who self-reported BLA were low risk fortype 1 hypersensitivity, but outpatient de-labelling efforts were largely unsuccessful.

Regulatory light chain phosphorylation and N-terminal extension increase cross-bridge binding and power output in Drosophila at in vivo myofilament lattice spacing.

The N-terminal extension and phosphorylation of the myosin regulatory light chain (RLC) independently improve Drosophila melanogaster flight performance. Here we examine the functional and structural role of the RLC in chemically skinned fibers at various thick and thin filament lattice spacings from four transgenic Drosophila lines: rescued null or control (Dmlc2(+)), truncated N-terminal extension (Dmlc2(Δ2-46)), disrupted myosin light chain kinase phosphorylation sites (Dmlc2(S66A,S67A)), and dual mutant (Dmlc2(Δ2-46; S66A,S67A)). The N-terminal extension truncation and phosphorylation sites disruption mutations decreased oscillatory power output and the frequency of maximum power output in maximally Ca(2+)-activated fibers compressed to near in vivo inter-thick filament spacing, with the phosphorylation sites disruption mutation having a larger affect. The diminished power output parameters with the N-terminal extension truncation and phosphorylation sites disruption mutations were due to the reduction of the number of strongly-bound cross-bridges and rate of myosin force production, with the larger parameter reductions in the phosphorylation sites disruption mutation additionally related to reduced myosin attachment time. The phosphorylation and N-terminal extension-dependent boost in cross-bridge kinetics corroborates previous structural data, which indicate these RLC attributes play a complementary role in moving and orienting myosin heads toward actin target sites, thereby increasing fiber and whole fly power generation.

COOH-terminal truncation of flightin decreases myofilament lattice organization, cross-bridge binding, and power output in Drosophila indirect flight muscle.

The indirect flight muscle (IFM) of insects is characterized by a near crystalline myofilament lattice structure that likely evolved to achieve high power output. In Drosophila IFM, the myosin rod binding protein flightin plays a crucial role in thick filament organization and sarcomere integrity. Here we investigate the extent to which the COOH terminus of flightin contributes to IFM structure and mechanical performance using transgenic Drosophila expressing a truncated flightin lacking the 44 COOH-terminal amino acids (fln(ΔC44)). Electron microscopy and X-ray diffraction measurements show decreased myofilament lattice order in the fln(ΔC44) line compared with control, a transgenic flightin-null rescued line (fln(+)). fln(ΔC44) fibers produced roughly 1/3 the oscillatory work and power of fln(+), with reduced frequencies of maximum work (123 Hz vs. 154 Hz) and power (139 Hz vs. 187 Hz) output, indicating slower myosin cycling kinetics. These reductions in work and power stem from a slower rate of cross-bridge recruitment and decreased cross-bridge binding in fln(ΔC44) fibers, although the mean duration of cross-bridge attachment was not different between both lines. The decreases in lattice order and myosin kinetics resulted in fln(ΔC44) flies being unable to beat their wings. These results indicate that the COOH terminus of flightin is necessary for normal myofilament lattice organization, thereby facilitating the cross-bridge binding required to achieve high power output for flight.

Obtaining a faculty position in STEM at a research-intensive institution.

Progressing from postdoctoral training to a STEM faculty appointment at a Research Intensive Institution (RII) is a daunting transition, and may be especially challenging to those who have followed a less-than-conventional path or whose peers have lost interest in academic careers. This article describes how to prepare for and progress through the application process for institutions in the USA, which takes approximately 1 year, including what to expect at each step and recommendations for a successful transition. The odds of success for any individual application are low, making good preparation and careful planning the more important, as does managing expectations to avoid becoming discouraged early in the process. The rewards of landing the faculty appointment at an institution that matches your professional and personal needs and for which you are best suited more than exceeds the effort required to attain it.

Secondary Structure of the Novel Myosin Binding Domain WYR and Implications within Myosin Structure.

Structural changes in the myosin II light meromyosin (LMM) that influence thick filament mechanical properties and muscle function are modulated by LMM-binding proteins. Flightin is an LMM-binding protein indispensable for the function of Drosophila indirect flight muscle (IFM). Flightin has a three-domain structure that includes WYR, a novel 52 aa domain conserved throughout Pancrustacea. In this study, we (i) test the hypothesis that WYR binds the LMM, (ii) characterize the secondary structure of WYR, and (iii) examine the structural impact WYR has on the LMM. Circular dichroism at 260-190 nm reveals a structural profile for WYR and supports an interaction between WYR and LMM. A WYR-LMM interaction is supported by co-sedimentation with a stoichiometry of ~2.4:1. The WYR-LMM interaction results in an overall increased coiled-coil content, while curtailing ɑ helical content. WYR is found to be composed of 15% turns, 31% antiparallel ß, and 48% 'other' content. We propose a structural model of WYR consisting of an antiparallel ß hairpin between Q92-K114 centered on an ASX or ß turn around N102, with a G1 bulge at G117. The Drosophila LMM segment used, V1346-I1941, encompassing conserved skip residues 2-4, is found to possess a traditional helical profile but is interpreted as having <30% helical content by multiple methods of deconvolution. This low helicity may be affiliated with the dynamic behavior of the structure in solution or the inclusion of a known non-helical region in the C-terminus. Our results support the hypothesis that WYR binds the LMM and that this interaction brings about structural changes in the coiled-coil. These studies implicate flightin, via the WYR domain, for distinct shifts in LMM secondary structure that could influence the structural properties and stabilization of the thick filament, scaling to modulation of whole muscle function.

Contiguity and Structural Impacts of a Non-Myosin Protein within the Thick Filament Myosin Layers.

Myosin dimers arranged in layers and interspersed with non-myosin densities have been described by cryo-EM 3D reconstruction of the thick filament in Lethocerus at 5.5 Å resolution. One of the non-myosin densities, denoted the 'red density', is hypothesized to be flightin, an LMM-binding protein essential to the structure and function of Drosophila indirect flight muscle (IFM). Here, we build upon the 3D reconstruction results specific to the red density and its engagement with the myosin coiled-coil rods that form the backbone of the thick filament. Each independent red density winds its way through the myosin dimers, such that it links four dimers in a layer and one dimer in a neighboring layer. This area in which three distinct interfaces within the myosin rod are contacted at once and the red density extends to the thick filament core is designated the "multiface". Present within the multiface is a contact area inclusive of E1563 and R1568. Mutations in the corresponding Drosophila residues (E1554K and R1559H) are known to interfere with flightin accumulation and phosphorylation in Drosophila. We further examine the LMM area in direct apposition to the red density and identified potential binding residues spanning up to ten helical turns. We find that the red density is associated within an expanse of the myosin coiled-coil that is unwound by the third skip residue and the coiled-coil is re-oriented while in contact with the red density. These findings suggest a mechanism by which flightin induces ordered assembly of myosin dimers through its contacts with multiple myosin dimers and brings about reinforcement on the level of a single myosin dimer by stabilization of the myosin coiled-coil.

Comparative biomechanics of thick filaments and thin filaments with functional consequences for muscle contraction.

The scaffold of striated muscle is predominantly comprised of myosin and actin polymers known as thick filaments and thin filaments, respectively. The roles these filaments play in muscle contraction are well known, but the extent to which variations in filament mechanical properties influence muscle function is not fully understood. Here we review information on the material properties of thick filaments, thin filaments, and their primary constituents; we also discuss ways in which mechanical properties of filaments impact muscle performance.

Cardiac myosin binding protein-C is essential for thick-filament stability and flexural rigidity.

Using atomic force microscopy, we examined the contribution of cardiac myosin binding protein-C (cMyBP-C) to thick-filament length and flexural rigidity. Native thick filaments were isolated from the hearts of transgenic mice bearing a truncation mutation of cMyBP-C (t/t) that results in no detectable cMyBP-C and from age-matched wild-type controls (+/+). Atomic force microscopy images of these filaments were evaluated with an automated analysis algorithm that identified filament position and shape. The t/t thick-filament length (1.48 +/- 0.02 microm) was significantly (P < 0.01) shorter than +/+ (1.56 +/- 0.02 microm). This 5%-shorter thick-filament length in the t/t was reflected in 4% significantly shorter sarcomere lengths of relaxed isolated cardiomyocytes of the t/t (1.97 +/- 0.01 microm) compared to +/+ (2.05 +/- 0.01 microm). To determine if cMyBP-C contributes to the mechanical properties of thick filaments, we used statistical polymer chain mechanics to calculate a per-filament-specific persistence length, an index of flexural rigidity directly proportional to Young's modulus. Thick-filament-specific persistence length in the t/t (373 +/- 62 microm) was significantly lower than in +/+ (639 +/- 101 microm). Accordingly, Young's modulus of t/t thick filaments was approximately 60% of +/+. These results provide what we consider a new understanding for the critical role of cMyBP-C in defining normal cardiac output by sustaining force and muscle stiffness.