Potent HPIV3-neutralizing IGHV5-51 Antibodies Identified from Multiple Individuals Show L Chain and CDRH3 Promiscuity.

Human parainfluenza virus 3 (HPIV3) is a widespread pathogen causing severe and lethal respiratory illness in at-risk populations. Effective countermeasures are in various stages of development; however, licensed therapeutic and prophylactic options are not available. The fusion glycoprotein (HPIV3 F), responsible for facilitating viral entry into host cells, is a major target of neutralizing Abs that inhibit infection. Although several neutralizing Abs against a small number of HPIV3 F epitopes have been identified to date, relatively little is known about the Ab response to HPIV3 compared with other pathogens, such as influenza virus and SARS-CoV-2. In this study, we aimed to characterize a set of HPIV3-specific Abs identified in multiple individuals for genetic signatures, epitope specificity, neutralization potential, and publicness. We identified 12 potently neutralizing Abs targeting three nonoverlapping epitopes on HPIV3 F. Among these, six Abs identified from two different individuals used Ig heavy variable gene IGHV 5-51, with five of the six Abs targeting the same epitope. However, despite the use of the same H chain variable (VH) gene, these Abs used multiple different L chain variable genes (VL) and diverse H chain CDR 3 (CDRH3) sequences. Together, these results provide further information about the genetic and functional characteristics of HPIV3-neutralizing Abs and suggest the existence of a reproducible VH-dependent Ab response associated with VL and CDRH3 promiscuity. Understanding sites of HPIV3 F vulnerability and the genetic and molecular characteristics of Abs targeting these sites will help guide efforts for effective vaccine and therapeutic development.

Combination of laser microdissection, 2D-DIGE and MALDI-TOF MS to identify protein biomarkers to predict colorectal cancer spread.

Biomarkers are urgently required to support current histological staging to provide additional accuracy in stratifying colorectal cancer (CRC) patients according to risk of spread to properly assign adjuvant chemotherapy after surgery. Chemotherapy is given to patients with stage III to reduce the risk of recurrence but is controversial in stage II patients. Up to 25% of stage II patients will relapse within 5 years after tumor removal and when this occurs cure is seldom possible. The aim of this study was to identify protein biomarkers to stratify risk of spread of CRC patients. Laser micro-dissection was used to isolate cancer cells from primary colorectal tumors of stage II patients which did or did not metastasize within 5 years after surgical resection. Protein expression differences between two groups of tumors were profiled by 2D-DIGE with saturation CyDye labeling and identified using MALDI-TOF mass spectrometry. Evaluation of protein candidates was conducted using tissue micro array (TMA) immunohistochemistry on 125 colorectal tumor tissue samples of different stages. A total of 55 differentially expressed proteins were identified. Ten protein biomarkers were chosen based on p value and ratio between non metastasized and metastazised groups and evaluated on 125 tissues using TMA immunohistochemistry. Expression of HLAB, protein 14-3-3ß, LTBP3, ADAMTS2, JAG2 and NME2 on tumour cells was significantly associated with clinical parameters related to tumour progression, invasion and metastasis. Kaplan-Meier survival curve showed strong expression of six proteins was associated with good CRC specific survival. Expression of HLAB, ADAMTS2, LTBP3, JAG2 and NME2 on tumour cells, was associated with tumour progression and invasion, metastasis and CRC specific survival may serve as potential biomarkers to stratify CRC patients into low and high risk of tumour metastasis. Combined methods of laser microdissection, 2D DIGE with saturation labelling and MALDI-TOF MS proved to be resourceful techniques capable of identifying protein biomarkers to predict risk of spread of CRC to liver.

Prevalence of Comorbid Conditions and Sun-Induced Skin Cancers in Patients with Alopecia Areata.

Alopecia areata is a multifactorial autoimmune disease causing non-scarring hair loss. Recent genome-wide association studies have pointed to connections between alopecia areata and other autoimmune disorders. Research of clinical conditions positively and negatively associated with alopecia areata is crucial for discovering the pathological mechanisms of disease and further treatment options.

Absorption of transdermal and oral cyclosporine in six healthy cats.

Cyclosporine is commonly used orally to treat feline dermatoses. Due to difficulties administering oral medications, veterinarians sometimes prescribe compounded transdermal cyclosporine, despite studies showing limited absorption. The study objective was to compare cyclosporine blood concentrations after oral administration to concentrations after transdermal application of cyclosporine (prepared in pluronic lecithin organogel [PLO]) in six cats using a controlled, cross-over design with a 2 wk washout period. Cats were dosed at 5.1-7.4 mg/kg of cyclosporine q 24 hr either per os for 7 days or transdermally for 21 days. Cyclosporine blood concentrations were measured q 7 days and after the washout period. A monoclonal-based immunoassay (lower limit of quantitation was 25 ng/mL) was used. Median concentrations on the seventh day were 2,208 ng/mL (range, 1,357-3,419 ng/mL) 2 hr after orally administered cyclosporine and 37 ng/mL (range, 25-290 ng/mL) 2 hr after transdermally applied cyclosporine. Median concentration on day 21 was 58 ng/mL (range, 51-878 ng/mL) 2 hr after transdermally applied cyclosporine. Concentrations were quantifiable for transdermally applied cyclosporine, but considered therapeutic in only one of six cats. Based on those results, transdermally applied cyclosporine was not recommended in cats because of inconsistent absorption.

Human monoclonal antibodies protect against viral-mediated pneumococcal superinfection.

Introduction: Community-acquired pneumonia (CAP) is a global health concern, with 25% of cases attributed to Streptococcus pneumoniae (Spn). Viral infections like influenza A virus (IAV), respiratory syncytial virus (RSV), and human metapneumovirus (hMPV) increase the risk of Spn, leading to severe complications due to compromised host immunity. Methods: We evaluated the efficacy of an anti-PhtD monoclonal antibody (mAb) cocktail therapy (PhtD3 + 7) in improving survival rates in three viral/bacterial coinfection models: IAV/Spn, hMPV/Spn, and RSV/Spn. Results: The PhtD3 + 7 mAb cocktail outperformed antiviral mAbs, resulting in prolonged survival. In the IAV/Spn model, it reduced bacterial titers in blood and lungs by 2-4 logs. In the hMPV/Spn model, PhtD3 + 7 provided greater protection than the hMPV-neutralizing mAb MPV467, significantly reducing bacterial titers. In the RSV/Spn model, PhtD3 + 7 offered slightly better protection than the antiviral mAb D25, uniquely decreasing bacterial titers in blood and lungs. Discussion: Given the threat of antibiotic resistance, our findings highlight the potential of anti-PhtD mAb therapy as an effective option for treating viral and secondary pneumococcal coinfections.

Structural basis for respiratory syncytial virus and human metapneumovirus neutralization.

Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) continue to be a global burden to infants, the elderly, and immunocompromised individuals. In the past ten years, there has been substantial progress in the development of new vaccine candidates and therapies against these viruses. These advancements were guided by the structural elucidation of the major surface glycoproteins for these viruses, the fusion (F) protein and attachment (G) protein. The identification of immunodominant epitopes on the RSV F and hMPV F proteins has expanded current knowledge on antibody-mediated immune responses, which has led to new approaches for vaccine and therapeutic development through the stabilization of pre-fusion constructs of the F protein and pre-fusion-specific monoclonal antibodies with high potency and efficacy. In this review, we describe structural characteristics of known antigenic sites on the RSV and hMPV proteins, their influence on the immune response, and current progress in vaccine and therapeutic development.

Gleason grading: past, present and future.

In 1966 Donald Gleason developed his grading and scoring system for prostatic adenocarcinoma. This classification was refined in 1974 and gained almost universal acceptance, being classified as a category 1 prognostic parameter by the College of American Pathologists. Modifications to the classification were recommended at a conference convened by the International Society of Urological Pathology (ISUP) in 2005. This modified classification has resulted in a significant upgrading of tumours, although some studies have shown a greater concordance between needle biopsy and radical prostatectomy scores when compared to classical Gleason (CG) grading. The ISUP consensus conference recommended that for needle biopsies higher tertiary patterns should be incorporated into the final Gleason score, and this has been correlated with biochemical failure, tumour volume and mortality. Recently the validity of including cribriform glands as a component of Gleason pattern 3 has been questioned and it has been recommended that all tumours showing cribriform architecture should be classified as Gleason pattern 4. The recommendations arising from the 2005 Consensus Conference were largely unsupported by validating data, yet this new grading system has achieved widespread usage. It is unfortunate that recent suggestions for further modification are similarly lacking in supporting evidence. In view of this it is recommended that the Modified Gleason Scoring Classification should continue to be utilized in its original (2005) format and that any future alterations should be implemented only when mandated by tumour-related outcome studies.

Side population is not necessary or sufficient for a cancer stem cell phenotype in glioblastoma multiforme.

There is strong evidence for the existence of cancer stem cells (CSCs) in the aggressive brain tumor glioblastoma multiforme (GBM). These cells have stem-like self-renewal activity and increased tumor initiation capacity and are believed to be responsible for recurrence due to their resistance to therapy. Several techniques have been used to enrich for CSC, including growth in serum-free defined media to induce sphere formation, and isolation of a stem-like cell using exclusion of the fluorescent dye Hoechst 33342, the side population (SP). We show that sphere formation in GBM cell lines and primary GBM cells enriches for a CSC-like phenotype of increased self-renewal gene expression in vitro and increased tumor initiation in vivo. However, the SP was absent from all sphere cultures. Direct isolation of the SP from the GBM lines did not enrich for stem-like activity in vitro, and tumor-initiating activity was lower in sorted SP compared with non-SP and parental cells. Transient exposure to doxorubicin enhanced both CSC and SP frequency. However, doxorubicin treatment altered the cytometric profile and obscured the SP demonstrating the difficulty of identifying SP in cells under stress. Doxorubicin-exposed cells showed a transient increase in SP, but the doxorubicin-SP cells were still not enriched for a stem-like self-renewal phenotype. These data demonstrate that the GBM SP does not necessarily contribute to self-renewal or tumor initiation, key properties of a CSC, and we advise against using SP to enumerate or isolate CSC.

A Pan-Pneumovirus vaccine based on immunodominant epitopes of the fusion protein.

Respiratory syncytial virus (RSV) and human metapneumovirus (hMPV) are two leading causes of severe respiratory infections in children, the elderly, and immunocompromised patients. The fusion (F) protein is the major target of neutralizing antibodies. Recent developments in stabilizing the pre-fusion conformation of the F proteins, and identifying immunodominant epitopes that elicit potent neutralizing antibodies have led to the testing of numerous pre-fusion RSV F-based vaccines in clinical trials. We designed and tested the immunogenicity and protective efficacy of a chimeric fusion protein that contains immunodominant epitopes of RSV F and hMPV F (RHMS-1). RHMS-1 has several advantages over vaccination with pre-fusion RSV F or hMPV F, including a focus on recalling B cells to the most important protective epitopes and the ability to induce protection against two viruses with a single antigen. RHMS-1 was generated as a trimeric recombinant protein, and analysis by negative-stain electron microscopy demonstrated the protein resembles the pre-fusion conformation. Probing of RHMS-1 antigenicity using a panel of RSV and hMPV F-specific monoclonal antibodies (mAbs) revealed the protein retains features of both viruses, including the pre-fusion site Ø epitope of RSV F. Mice immunized with RHMS-1 generated neutralizing antibodies to both viruses and were completely protected from RSV or hMPV challenge. Overall, this study demonstrates protection against two viruses with a single antigen and supports testing of RHMS-1 in additional pre-clinical animal models.

Future Well-Being Among U.S. Youth Who Attempted Suicide and Survived.

To what extent does a suicide attempt impair a person's future well-being? We estimated the prevalence of future well-being (FWB) among suicide attempt survivors using a nationally representative sample of 15,170 youths. Suicide attempt survivors were classified as having high FWB if they reported (a) a suicide attempt at Wave I, (b) no suicidal ideation or attempts over the past year at Wave III (7 years after), and (c) a well-being profile at or above the top quartile of nonsuicidal peers. Seventy-five of 574 suicide attempt survivors (∼13%) met criteria for FWB at Wave III, compared to 26% of nonsuicidal peers. Wave I well-being levels, not depressive symptoms, predicted the likelihood of FWB at Wave III (OR = 1.23, 95% CI [1.05, 1.44], p < .05). In conclusion, a nonfatal suicide attempt reduced but did not preclude FWB in a large national sample. The observation that a segment of the population of suicide attempt survivors achieves FWB carries implications for the prognosis of suicidal behavior and the value of incorporating well-being into investigations of suicide-related phenomena.

Only Human: Mental-Health Difficulties Among Clinical, Counseling, and School Psychology Faculty and Trainees.

How common are mental-health difficulties among applied psychologists? This question is paradoxically neglected, perhaps because disclosure and discussion of these experiences remain taboo within the field. This study documented high rates of mental-health difficulties (both diagnosed and undiagnosed) among faculty, graduate students, and others affiliated with accredited doctoral and internship programs in clinical, counseling, and school psychology. More than 80% of respondents (n = 1,395 of 1,692) reported a lifetime history mental-health difficulties, and nearly half (48%) reported a diagnosed mental disorder. Among those with diagnosed and undiagnosed mental-health difficulties, the most common reported concerns were depression, generalized anxiety disorder, and suicidal thoughts or behaviors. Participants who reported diagnosed mental disorders endorsed, on average, more specific mental-health difficulties and were more likely to report current difficulties than were undiagnosed participants. Graduate students were more likely to endorse both diagnosed and undiagnosed mental-health difficulties than were faculty, and they were more likely to report ongoing difficulties. Overall, rates of mental disorders within clinical, counseling, and school-psychology faculty and trainees were similar to or greater than those observed in the general population. We discuss the implications of these results and suggest specific directions for future research on this heretofore neglected topic.

Observations of supermicron-sized aerosols originating from biomass burning in southern Central Africa.

During the 3 years of the ObseRvations of Aerosols above CLouds and their intEractionS (ORACLES) campaign, the NASA Orion P-3 was equipped with a 2D stereo (2D-S) probe that imaged particles with maximum dimension (D) ranging from 10 < D < 1280 µm. The 2D-S recorded supermicron-sized aerosol particles (SAPs) outside of clouds within biomass burning plumes during flights over the southeastern Atlantic off Africa's coast. Numerous SAPs with 10 < D < 1520 µm were observed in 2017 and 2018 at altitudes between 1230 and 4000 m, 1000 km from the coastline, mostly between 7-11° S. No SAPs were observed in 2016 as flights were conducted further south and further from the coastline. Number concentrations of refractory black carbon (rBC) measured by a single particle soot photometer ranged from 200 to 1200 cm-3 when SAPs were observed. Transmission electron microscopy images of submicron particulates, collected on Holey carbon grid filters, revealed particles with potassium salts, black carbon (BC), and organics. Energy-dispersive X-ray spectroscopy spectra also detected potassium, a tracer for biomass burning. These measurements provided evidence that the submicron particles originated from biomass burning. NOAA Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT) 3 d back trajectories show a source in northern Angola for times when large SAPs were observed. Fire Information for Resource Management System (FIRMS) Moderate Resolution Imaging Spectroradiometer (MODIS) 6 active fire maps showed extensive biomass burning at these locations. Given the back trajectories, the high number concentrations of rBC, and the presence of elemental tracers indicative of biomass burning, it is hypothesized that the SAPs imaged by the 2D-S are examples of BC aerosol, ash, or unburned plant material.

Institutional Interventional Radiology Symposium Increases Medical Student Interest and Identifies Target Recruitment Candidates.

OBJECTIVES: To assess and raise medical student interest in interventional radiology (IR); and to evaluate student response across gender, level of training, and surgical vs nonsurgical specialty interest. MATERIALS AND METHODS: All Ohio medical students were invited to an IR Symposium held by a large academic medical center in central Ohio. The program encompassed didactic lectures, hands-on simulation models, and a networking luncheon with faculty, trainees, and industry partners. All attendees completed an anonymous, 5-point Likert scaled survey preattending and postattending the event to assess their awareness of IR as a specialty, understanding of the current training pathways, and level of interest. RESULTS: A total of 46 participants (M:F 60%:40%, MS1-53%, MS2-36%, and MS3-11%) attended the symposium. The cohort demonstrated increased interest in pursuing a career in IR following the symposium (4.12 vs 3.70, P < 0.001). Students with an interest in a nonsurgical specialty showed an increased interest in IR (4.20 vs 3.68, P < 0.001), whereas surgically oriented students did not demonstrate a significant increase (4.00 vs 3.71, P = 0.375). No statistically significant differences were noted across gender or level of training. The symposium experience significantly increased understanding of the IR training pathways (4.51 vs 2.94, P < 0.001). Students rated lectures (57%) and endovascular simulators (41%) as the most useful experiences. CONCLUSIONS: This study demonstrated the role of symposia in improving medical student awareness of IR and training pathways. Findings were validated across gender and training level, and identified the subset of students with nonsurgical interests as most responsive to such intervention and potential recruitment.

Implementation of an Innovative Tablet-based Curriculum for Radiology Resident Education.

RATIONALE AND OBJECTIVES: The aim of this study was to prospectively examine the impact of a tablet-based curriculum on the radiology resident educational experience. MATERIALS AND METHODS: A comprehensive tablet-based curriculum was developed by creating subspecialty modules with appropriate content level for each required rotation at our diagnostic radiology residency program. Daily assignments included key learning points, readings, and reference presentation slides, covering all objectives published by the ABR Core Exam Study Guide. Residents were provided with iPad devices preconfigured with the curriculum and online access to most major radiology textbooks available in our institutional digital library. Assessment surveys were administered at baseline and 12 months following curriculum implementation. RESULTS: Twenty-two residents completed both surveys. In comparing the pre versus postsurvey results, 32% versus 73% residents agreed or strongly agreed that study resources were well-organized, 41% versus 91% agreed or strongly agreed that study resources were easily accessible, 27% versus 77% agreed or strongly agreed that the modules encouraged active learning, 18% versus 82% agreed or strongly agreed that resources motivate them to study daily, 36% versus 82% agreed or strongly agreed that the resources adequately prepared the resident for the radiology board exam, and 36% versus 82% agreed or strongly agreed that they were satisfied with the resources provided by the residency program (p < 0.05). CONCLUSION: Our study demonstrated the positive impact of implementing a complete tablet-based curriculum on radiology resident motivation, satisfaction, and engagement. Use of mobile tablet devices has the potential to dramatically transform content delivery in residency education.

Competition for dead trees between humans and aye-ayes (Daubentonia madagascariensis) in central eastern Madagascar.

The destruction and degradation of forest habitats are major threats to the sustainability of lemur populations in Madagascar. Madagascan landscapes often contain forest fragments that represent refuges for native fauna, while also being used for firewood and timber by local human populations. As undisturbed forest becomes increasingly scarce, understanding resource competition between humans and wildlife in disturbed habitats will be increasingly important. We tested the hypothesis that Malagasy and aye-ayes (Daubentonia madagascariensis) compete for the limited number of dead trees in rainforest fragments at Tsinjoarivo, Madagascar. We surveyed 2.16 ha within five fragments (range 5-228 ha) surrounding human settlements to quantify the density of dead trees and traces of both human and aye-aye activity. Neither aye-aye nor human traces were distributed according to the availability of particular trees species, and aye-ayes and Malagasy apparently preferred several different species. Although overlap was recorded in tree species used, human use tended to be positively correlated with a species' desirability as firewood, while a negative relationship was seen for aye-ayes. Both consumers used trees of similar diameter at breast height, but those used by aye-ayes tended to be older, suggesting that human use might precede usefulness for aye-ayes. Finally, the density of dead trees and aye-aye traces were highest in smaller fragments, but human traces did not vary across fragment size. Although further study is needed to better quantify the aye-aye diet in this region, these data suggest that aye-ayes and local people compete for dead trees, and this competition could constitute a pressure on aye-aye populations.

Executive Functioning and Engagement in Physical and Relational Aggression among Children with ADHD.

Although evidence suggests that executive functioning (EF) impairments are implicated in physically aggressive behavior (e.g., hitting) these cognitive impairments have rarely been examined with regard to relational aggression (e.g., gossip, systematic exclusion). Studies also have not examined if EF impairments underlie the expression of aggression in children with attention-deficit/hyperactivity disorder (ADHD) and if child gender moderates risk. Children with and without clinical elevations in ADHD symptoms (N = 124; ages 8-12 years; 48 % male) completed a battery of EF tests. Parent and teacher report of ADHD and oppositional defiant disorder (ODD) symptoms and teacher report of engagement in physical and relational aggression were collected. Models tested the unique association of EF abilities with physical and relational aggression and the indirect effect through the expression of ADHD or ODD behaviors; child gender was also tested as a moderator. EF impairment was uniquely associated with physical aggression, but better EF ability was associated with relational aggression. For boys, poor EF also was indirectly associated with greater physical aggression through the expression of ADHD behaviors. However, ADHD symptoms were unrelated to relational aggression. ODD symptoms also predicted physical aggression for boys but relational aggression for girls. Results suggest that there are multiple and distinct factors associated with engagement in physical and relational aggression and that better EF may actually promote relational aggression. Established models of physical aggression should not be assumed to map on to explanations of relational aggression.

Canine alopecia secondary to human topical hormone replacement therapy in six dogs.

Alopecia is a common presenting complaint in veterinary medicine and is known to occur secondary to numerous primary conditions. In this report, six unrelated dogs from three households were subsequently determined to have developed alopecia as a result of accidental transdermal exposure to their owners' topical hormone replacement therapy (THRT). All cases presented with alopecia ranging in duration from 2 mo to 2.5 yr. All dogs demonstrated alopecia affecting the ventral neck, thoracic and abdominal surfaces, proximal lateral extremities, and lateral trunk. At the time of initial presentation, five of six dogs were also noted to have physical exam findings suggestive of feminization. In all cases, serum total thyroxine was within normal reference range. Affected skin was biopsied in five dogs, and all samples demonstrated four similar histological characteristics: basal melanosis, epidermal and infundibular follicular hyperkeratosis, kenogen hair follicles, and small sebaceous glands. All dogs had elevated baseline estradiol levels, and four dogs had concurrent elevations of baseline progesterone. Average time to onset of clinical signs in those dogs was 5.5 mo after the owners started THRT. Following discontinuation of THRT by the owners, all dogs had complete resolution of their clinical signs by 5.5 mo.

Pathologies of the appendix: a 10-year review of 4670 appendicectomy specimens.

BACKGROUND: Debate surrounds the management of the macroscopically normal appendix. Current literature recommends its removal given the high incidence of microscopic appendicitis, and other unusual pathologies in the normal-looking appendix. Negative appendicectomies are reported on the decline with increased use of diagnostic radiological adjuncts. METHODS: This study analysed pathologies of the appendix over 10 years in the Pathology Department in Canberra. A positive appendicectomy was defined as acute appendicitis, faecoliths, worms, endometriosis or appendiceal tumours. We reviewed the positive appendicectomy rate over this time period. RESULTS: There were 4670 appendicectomy specimens in 2386 males (51.1%) and 2284 (49%) females. The incidence of acute appendicitis was 71.3% and the positive appendicectomy rate was 76.3%. There were significantly fewer negative appendicectomies in males (16.8%) compared with females (31.0%). There was no appreciable change in this trend over the study period. Of the positive appendicectomies, there were 129 (3.6%) faecoliths. Of these, only 39.5% had concomitant appendicitis. There were 44 (1.2%) specimens identified with worms. Of these, 40.9% had concomitant appendicitis. There were 14 cases of endometriosis of the appendix of which 36% had concomitant appendicitis. There were 58/3562 (1.6%) appendiceal tumours within the positive appendicectomy group the majority of which were carcinoid tumours (65.5%). CONCLUSION: There is a higher incidence of negative appendicectomies in women compared with men, which is similar to other published studies. Faecoliths and worms are a known cause of appendiceal colic and in our series were identified mostly in the absence of histological evidence of appendicitis.