Improved intraoperative identification of close margins in oral squamous cell carcinoma resections using a dual aperture fluorescence ratio approach: first in-human results.

Significance: Surgical excision is the main treatment for solid tumors in oral squamous cell carcinomas, where wide local excision (achieving a healthy tissue margin of >5 mm around the excised tumor) is the goal as it results in reduced local recurrence rates and improved overall survival. Aim: No clinical methods are available to assess the complete surgical margin intraoperatively while the patient is still on the operating table; and while recent intraoperative back-bench fluorescence-guided surgery approaches have shown promise for detecting "positive" inadequate margins (<1 mm), they have had limited success in the detection of "close" inadequate margins (1 to 5 mm). Here, a dual aperture fluorescence ratio (dAFR) approach was evaluated as a means of improving detection of close margins. Approach: The approach was evaluated on surgical specimens from patients who were administered a tumor-specific fluorescent imaging agent (cetuximab-800CW) prior to surgery. The dAFR approach was compared directly against standard wide-field fluorescence imaging and pathology measurements of margin thickness in specimens from three patients and a total of 12 margin locations (1 positive, 5 close, and 6 clear margins). Results: The area under the receiver operating characteristic curve, representing the ability to detect close compared to clear margins (>5 mm) was found to be 1.0 and 0.57 for dAFR and sAF, respectively. Improvements in dAFR were found to be statistically significant (p<0.02). Conclusions: These results provide evidence that the dAFR approach potentially improves detection of close surgical margins.

C9orf72 proline-arginine dipeptide repeats disrupt the proteasome and perturb proteolytic activities.

The hexanucleotide G4C2 repeat expansion in C9orf72 is the most frequent genetic cause of familial amyotrophic lateral sclerosis (ALS). Aberrant translation of this hexanucleotide sequence leads to production of 5 dipeptide repeats (DPRs). One of these DPRs is proline-arginine (polyPR), which is found in C9orf72-expanded ALS (C9ALS) patient brain tissue and is neurotoxic across multiple model systems. PolyPR was previously reported to bind and impair proteasomes in vitro. Nevertheless, the clinical relevance of the polyPR-proteasome interaction and its functional consequences in vivo are yet to be established. Here, we aim to confirm and functionally characterize polyPR-induced impairment of proteolysis in C9ALS patient tissue and an in vivo model system. Confocal microscopy and immunofluorescence studies on both human and Drosophila melanogaster brain tissues revealed sequestration of proteasomes by polyPR into inclusion-like bodies. Co-immunoprecipitation in D. melanogaster showed that polyPR strongly binds to the proteasome. In vivo, functional evidence for proteasome impairment is further shown by the accumulation of ubiquitinated proteins along with lysosomal accumulation and hyper-acidification, which can be rescued by a small-molecule proteasomal enhancer. Together, we provide the first clinical report of polyPR-proteasome interactions and offer in vivo evidence proposing polyPR-induced proteolytic dysfunction as a pathogenic mechanism in C9ALS.

Underrepresentation of women in exercise science and physiology research is associated with authorship gender.

Historically, low representation of women participants in exercise science and physiology studies has led to a lack of understanding in the response of women to exercise and therapeutic interventions. We hypothesized that 1) the number of women authors, participants, and editorial board members increased over 30 years (1991-2021) and 2) larger representation of women as editors and authors is associated with more women participants. Gender (man/woman) of editorial board members (n = 394), authors (n = 5,735), and participants (n = 2,984,883) of 972 original research articles with human participants published in 1991 and 2021 was analyzed from three journals: Journal of Applied Physiology, Medicine and Science in Sports and Exercise, and British Journal of Sports Medicine. Between 1991 to 2021, the average percent women per article as participants (21.9 ± 31.7% vs. 36.3 ± 30.3%, respectively, P < 0.001), authors (16.4 ± 22.4% vs. 30.9 ± 24.0%, P < 0.001), and editorial board members (13.3 ± 5.4% vs. 41.5 ± 7.3%, P = 0.006) increased. In 2021, the gender proportion of participants in large datasets was similar (50.2 ± 20.2% women). However, studies with smaller datasets (i.e., <∼3,000 participants) included less women (35.6 ± 30.6%). Women participants (%) were less when the last author was a man rather than a woman in 1991 (19.9 ± 29.5% vs. 34.3 ± 42.2%) and 2021 (31.6 ± 27.7% vs. 51.7 ± 33.4%). In 2021, there was a positive correlation between author and participant gender (% women) (r = 0.42, P < 0.001). Our data suggest that the low representation of women in exercise science and physiology research could be resolved with equitable numbers of women authors and editors and by encouraging men authors to study both women and men participants.NEW & NOTEWORTHY Analysis of human applied physiology studies revealed that the representation of women authors, participants, and editorial board members increased over 30 years but remained lower than men in 2021. Larger representation of women editors and authors was associated with more women participants. Women authors assessed similar numbers of women and men participants, whereas men authors included less women. Equitable representation of women participants may be achieved by closing the gender gap in authorship and editorial board membership.