Two modes of transvection at the eyes absent gene of Drosophila demonstrate plasticity in transcriptional regulatory interactions in cis and in trans.

For many genes, proper gene expression requires coordinated and dynamic interactions between multiple regulatory elements, each of which can either promote or silence transcription. In Drosophila, the complexity of the regulatory landscape is further complicated by the tight physical pairing of homologous chromosomes, which can permit regulatory elements to interact in trans, a phenomenon known as transvection. To better understand how gene expression can be programmed through cis- and trans-regulatory interactions, we analyzed transvection effects for a collection of alleles of the eyes absent (eya) gene. We find that trans-activation of a promoter by the eya eye-specific enhancers is broadly supported in many allelic backgrounds, and that the availability of an enhancer to act in trans can be predicted based on the molecular lesion of an eya allele. Furthermore, by manipulating promoter availability in cis and in trans, we demonstrate that the eye-specific enhancers of eya show plasticity in their promoter preference between two different transcriptional start sites, which depends on promoter competition between the two potential targets. Finally, we show that certain alleles of eya demonstrate pairing-sensitive silencing resulting from trans-interactions between Polycomb Response Elements (PREs), and genetic and genomic data support a general role for PcG proteins in mediating transcriptional silencing at eya. Overall, our data highlight how eya gene regulation relies upon a complex but plastic interplay between multiple enhancers, promoters, and PREs.

Mass spectrometry-based molecular mapping of native FXIIIa cross-links in insoluble fibrin clots.

The roles of factor XIIIa-specific cross-links in thrombus formation, regression, or probability for embolization are largely unknown. A molecular understanding of fibrin architecture at the level of these cross-links could inform the development of therapeutic strategies to prevent the sequelae of thromboembolism. Here, we present an MS-based method to map native factor XIIIa cross-links in the insoluble matrix component of whole-blood or plasma-fibrin clots and in in vivo thrombi. Using a chaotrope-insoluble digestion method and quantitative cross-linking MS, we identified the previously mapped fibrinogen peptides that are responsible for covalent D-dimer association, as well as dozens of novel cross-links in the αC region of fibrinogen α. Our findings expand the known native cross-linked species from one to over 100 and suggest distinct antiparallel registries for interprotofibril association and covalent attachment of serpins that regulate clot dissolution.

Characteristics of hospitalizations among patients receiving immune checkpoint inhibitors at a community teaching hospital.

PURPOSE: As immune checkpoint inhibitors continue to acquire new indications, it is important to understand the impact their use has on patients. This study adds to current literature by presenting an analysis of hospitalizations in this population. The primary objective was to assess the reasons for an emergency department visit or hospital admission in patients who receive immune checkpoint inhibitors. Secondary objectives included identifying the frequency of suspected or confirmed immune related adverse events, types of immune related adverse events, number of preventable admissions, duration of immunotherapy, and length of stay. METHODS: This study was a retrospective, multi-center, chart review of patients hospitalized after receiving an immune checkpoint inhibitor. The population included patients aged 18 and above who received at least one dose of an immune checkpoint inhibitor at a network facility and had a documented admission within one year following the initiation of immunotherapy. Descriptive statistics were performed along with inferential comparisons and a Poisson regression to determine if the immune checkpoint blocker or cancer type predicted admission or reason for admission. RESULTS: The 99 patients who met inclusion criteria had a total of 202 admissions. Of these patients, 56 (56.6%) had multiple admissions within the year following initiation of immunotherapy. The most common diagnoses on initial admissions were shortness of breath, pain, and pneumonia. A total of 104 admissions (51.5%) were considered potentially preventable. Suspected or confirmed immune related adverse events were identified in 15.6% of all admissions. There were no significant predictors of admissions or reason for admission. CONCLUSION: Reasons for admission in the study population were comparable to those identified in the general cancer population, with immune related adverse events being associated with a minority of both total and potentially preventable admissions.

Enhancing research for undergraduates through a nanotechnology training program that utilizes analytical and bioanalytical tools.

Nanotechnology is a broad field combining traditional scientific disciplines; however, analytical chemistry plays an important role in material design, synthesis, characterization, and application. This article emphasizes the uniqueness of nanotechnology and the importance of providing high-quality undergraduate research experiences to both attract and retain talented individuals to the field of nanotechnology. In response to this need to develop a strong and sustainable nanotechnology work force, strategies to create authentic research experiences are considered within the framework of an interdisciplinary nanotechnology environment at West Virginia University. The program, named NanoSAFE Research Experiences for Undergraduates (REU), embeds students in different departments at West Virginia University and in research laboratories within the National Institute of Occupational Safety and Health. A large number of participants have little or no prior research experience and a strong effort is made to recruit applicants from under-represented populations. Components designed to foster research proficiency include frequent reporting, a strong peer-network, and training for secondary mentors. Evidence, which includes student publications and assessment findings demonstrating self-efficacy, is discussed to substantiate the viability of the strategies used in the 2016-2018 program. Graphical abstract ᅟ.

Knowledge and practices related to louse- and flea-borne diseases among staff providing services to people experiencing homelessness in the United States.

BACKGROUND AND AIMS: Louse-borne Bartonella quintana infection and flea-borne murine typhus are two potentially serious vector-borne diseases that have led to periodic outbreaks among people experiencing homelessness in the United States. Little is known about louse- and flea-borne disease awareness and prevention among staff who provide services to the population. We surveyed staff in seven US states to identify gaps in knowledge and prevention practices for these diseases. METHODS AND RESULTS: Surveys were administered to 333 staff at 89 homeless shelters and outreach teams in California, Colorado, Georgia, Maryland, Minnesota, New York and Washington from August 2022 to April 2023. Most participants (>68%) agreed that body lice and fleas are a problem for people experiencing homelessness. About half were aware that diseases could be transmitted by these vectors; however, most could not accurately identify which diseases. Less than a quarter of staff could describe an appropriate protocol for managing body lice or fleas. Misconceptions included that clients must isolate or be denied services until they are medically cleared. CONCLUSIONS: Our findings reveal significant knowledge gaps among staff who provide services to people experiencing homelessness in the prevention and control of louse- and flea-borne diseases. This demonstrates an urgent need for staff training to both reduce disease and prevent unnecessary restrictions on services and housing.

Effects of audio-visual integration on the detection of masked speech and non-speech sounds.

Integration of simultaneous auditory and visual information about an event can enhance our ability to detect that event. This is particularly evident in the perception of speech, where the articulatory gestures of the speaker's lips and face can significantly improve the listener's detection and identification of the message, especially when that message is presented in a noisy background. Speech is a particularly important example of multisensory integration because of its behavioural relevance to humans and also because brain regions have been identified that appear to be specifically tuned for auditory speech and lip gestures. Previous research has suggested that speech stimuli may have an advantage over other types of auditory stimuli in terms of audio-visual integration. Here, we used a modified adaptive psychophysical staircase approach to compare the influence of congruent visual stimuli (brief movie clips) on the detection of noise-masked auditory speech and non-speech stimuli. We found that congruent visual stimuli significantly improved detection of an auditory stimulus relative to incongruent visual stimuli. This effect, however, was equally apparent for speech and non-speech stimuli. The findings suggest that speech stimuli are not specifically advantaged by audio-visual integration for detection at threshold when compared with other naturalistic sounds.

Altered active control of step width in response to mediolateral leg perturbations while walking.

During human walking, step width is predicted by mediolateral motion of the pelvis, a relationship that can be attributed to a combination of passive body dynamics and active sensorimotor control. The purpose of the present study was to investigate whether humans modulate the active control of step width in response to a novel mechanical environment. Participants were repeatedly exposed to a force-field that either assisted or perturbed the normal relationship between pelvis motion and step width, separated by washout periods to detect the presence of potential after-effects. As intended, force-field assistance directly strengthened the relationship between pelvis displacement and step width. This relationship remained strengthened with repeated exposure to assistance, and returned to baseline afterward, providing minimal evidence for assistance-driven changes in active control. In contrast, force-field perturbations directly weakened the relationship between pelvis motion and step width. Repeated exposure to perturbations diminished this negative direct effect, and produced larger positive after-effects once the perturbations ceased. These results demonstrate that targeted perturbations can cause humans to adjust the active control that contributes to fluctuations in step width.

Ultrasound-Guided First Dorsal Compartment Release for Refractory de Quervain Tenosynovitis: A Case Report.

De Quervain tenosynovitis is an overuse syndrome associated with inflammation of the tendons in the first dorsal compartment of the wrist. Management includes activity modification, splinting, and corticosteroid injections. In refractory cases, surgical release may be performed, which may be complicated by incomplete release, tendon subluxation, or radial nerve injury. Alternatively, ultrasound-guided release may improve patient satisfaction and outcomes through faster recovery time as well as improved visualization of subcompartments and neurovascular structures. To the best of the authors' knowledge, this is the first case in the literature describing a novel technique for ultrasound-guided first dorsal compartment release for refractory de Quervain tenosynovitis.

Redetermination of conichalcite, CaCu(AsO(4))(OH).

The crystal structure of conichalcite [calcium copper(II) arsenate(V) hydroxide], with ideal formula CaCu(AsO(4))(OH), was redetermined from a natural twinned specimen found in the Maria Catalina mine (Chile). In contrast to the previous refinement from photographic data [Qurashi & Barnes (1963 ▶). Can. Mineral. 7, 561-577], all atoms were refined with anisotropic displacement parameters and with the H atom located. Conichalcite belongs to the adelite mineral group. The Jahn-Teller-distorted [CuO(6)] octa-hedra share edges, forming chains running parallel to [010]. These chains are cross-linked by eight-coordinate Ca atoms and by sharing vertices with isolated AsO(4) tetra-hedra. Of five calcium arsenate minerals in the adelite group, the [MO(6)] (M = Cu, Zn, Co, Ni and Mg) octa-hedron in conichalcite is the most distorted, and the donor-acceptor O-H⋯O distance is the shortest.

Pure bone marrow aspirate injection for chronic greater trochanteric pain syndrome: a case report.

There are limited treatment options for patients with a chronic refractory greater trochanteric pain syndrome. Orthobiologic interventions may stimulate tendon healing and improve pain and function in patients who fail the standard conservative treatment. Since the US FDA's new position statement regarding the centrifugation of bone marrow aspirate products as a potentially 'more than minimally manipulated' product, there is a growing concern about the most common bone marrow aspirate concentrate technique. In this case, a 57 year old female with a debilitating chronic greater trochanteric pain syndrome was treated with a pure autologous bone marrow aspirate injection using a novel aspiration technique. The patient showed significant improvements in pain and function without recurrence at 1-year follow-up. This is the first case report to illustrate this novel technique for aspirating pure bone marrow that should comply with the new FDA regulations.

Zebrafish blastomere screen identifies retinoic acid suppression of MYB in adenoid cystic carcinoma.

Pluripotent cells have been used to probe developmental pathways that are involved in genetic diseases and oncogenic events. To find new therapies that would target MYB-driven tumors, we developed a pluripotent zebrafish blastomere culture system. We performed a chemical genetic screen and identified retinoic acid agonists as suppressors of c-myb expression. Retinoic acid treatment also decreased c-myb gene expression in human leukemia cells. Translocations that drive overexpression of the oncogenic transcription factor MYB are molecular hallmarks of adenoid cystic carcinoma (ACC), a malignant salivary gland tumor with no effective therapy. Retinoic acid agonists inhibited tumor growth in vivo in ACC patient-derived xenograft models and decreased MYB binding at translocated enhancers, thereby potentially diminishing the MYB positive feedback loop driving ACC. Our findings establish the zebrafish pluripotent cell culture system as a method to identify modulators of tumor formation, particularly establishing retinoic acid as a potential new effective therapy for ACC.

Diagnosis and Treatment of Gouty Tophi in the Patellar Tendon Using Ultrasound-Guided Needle Barbotage: A Case Presentation.

A 73-year-old man presented with nontraumatic debilitating anterior left knee pain. Magnetic resonance imaging suggested a partial patellar tendon tear. However, ultrasound imaging revealed multiple small hyperechoic areas with posterior acoustic shadowing with intermixed hypoechoic fluid within the proximal patellar tendon, consistent with gouty tophi. The patient underwent ultrasound-guided needle barbotage of the tophi. Gout was confirmed by the presence of uric acid crystals in the aspirate. He had immediate pain relief. Ultrasound imaging 6 weeks later demonstrated healing of the tendon without tophi recurrence. At 6-month follow-up, the patient remained asymptomatic. This case highlights a rare cause of knee pain, promotes diagnostic ultrasound, and describes a novel treatment technique for intratendinous tophaceous gout. LEVEL OF EVIDENCE: V.

Long-term drug administration in the adult zebrafish using oral gavage for cancer preclinical studies.

Zebrafish are a major model for chemical genetics, and most studies use embryos when investigating small molecules that cause interesting phenotypes or that can rescue disease models. Limited studies have dosed adults with small molecules by means of water-borne exposure or injection techniques. Challenges in the form of drug delivery-related trauma and anesthesia-related toxicity have excluded the adult zebrafish from long-term drug efficacy studies. Here, we introduce a novel anesthetic combination of MS-222 and isoflurane to an oral gavage technique for a non-toxic, non-invasive and long-term drug administration platform. As a proof of principle, we established drug efficacy of the FDA-approved BRAF(V600E) inhibitor, Vemurafenib, in adult zebrafish harboring BRAF(V600E) melanoma tumors. In the model, adult casper zebrafish intraperitoneally transplanted with a zebrafish melanoma cell line (ZMEL1) and exposed to daily sub-lethal dosing at 100 mg/kg of Vemurafenib for 2 weeks via oral gavage resulted in an average 65% decrease in tumor burden and a 15% mortality rate. In contrast, Vemurafenib-resistant ZMEL1 cell lines, generated in culture from low-dose drug exposure for 4 months, did not respond to the oral gavage treatment regimen. Similarly, this drug treatment regimen can be applied for treatment of primary melanoma tumors in the zebrafish. Taken together, we developed an effective long-term drug treatment system that will allow the adult zebrafish to be used to identify more effective anti-melanoma combination therapies and opens up possibilities for treating adult models of other diseases.

Low-dose oral interferon modulates expression of inflammatory and autoimmune genes in cattle.

While the safety and efficacy profiles of orally administered bovine interferon (IFN) alpha have been documented, the mechanism(s) that result in clinical benefits remain elusive. One approach to delineating the molecular pathways of IFN efficacy is through the use of gene expression profiling technologies. In this proof-of-concept study, different (0, 50, 200 and 800 units) oral doses of natural bovine IFN (type I) were tested in cattle to determine if oral IFN altered the expression of genes that may be pivotal to the development of systemic resistance to viral infections such as foot-and-mouth disease (FMD). Oral IFN was administered twice: Time 0 and 8h later. Blood was collected at 0, 8 and 24h after the first IFN administration, and DNA isolated from peripheral blood mononuclear cells (PBMCs) was employed in quantitative polymerase chain reaction (qPCR) microarray assays. Within 8h, 50 and 200 units of oral IFN induced significant (P<0.05) changes in expression of 41 of 92 tested autoimmune and inflammatory response-associated genes. These data suggest that orally administered IFN is a viable approach for providing short-term antiviral immunity to livestock exposed to viruses such as FMD virus (FMDV) until such a time that an effective vaccine can be produced and distributed to producers.

Timing and location of rhodopsin expression in newly born rod photoreceptors in the adult teleost retina.

Labeling of newly divided retinal cells with bromodeoxyuridine (BrdU) and a rhodopsin mRNA probe revealed that rhodopsin is first expressed by new rod photoreceptors 2 days after cell birth in an adult cichlid fish. Most new cells that expressed rhodopsin had nuclei located in the vitreal half of the outer nuclear layer (ONL), lending further support to the hypothesis that movement from scleral to vitreal ONL is associated with rod differentiation.