Subsequent Time Points Research Articles

The Cellular Impact, Immediate Targets and Mechanism of Action of NSD2 in t(4;14) Multiple Myeloma

INTRODUCTION: NSD2 is a potential therapeutic target in the 15% of cases of multiple myeloma that carry a t(4;14) translocation. As a consequence of the translocation, strong Ig enhancers are placed in proximity to NSD2 and FGFR3. FGFR3 is deleted at a subsequent time point in 25% of cases, while all cases retain NSD2 expression consistent with it likely being the key oncogenic driver. NSD2 is an H3K36 dimethyl transferase, a histone modification mark associated with active transcription, but the molecular targets and the mechanism by which NSD2 regulates its targets is unknown. The cellular function of NSD2 is also largely unknown, but previous shRNA knockdown studies have suggested it is involved in cell proliferation. The major limitation to understanding the role of NSD2 is the lack of an appropriate experimental system with which to investigate its molecular and cellular function. To address this deficit and to identify the primary direct targets of NSD2 as a means to elucidate its mechanism of action we have generated NSD2 dTAG and NSD2 KO cells together with NSD2 reconstitution systems. METHODS: We have utilized CRISPR gene editing to knock-in a dTAG in NSD2 in the t(4;14) KMS-11 cell line. This system enables the inducible and rapid degradation of NSD2 following the addition of a small-molecule degrader which is otherwise inert. We have also generated clonal NSD2 KO KMS-11 cells that allow us to observe and study the stable phenotypes of NSD2 KO. Because H3K36me2 deposited by NSD2 leads to induction of DNMT3A-mediated DNA methylation and suppression of PRC2-mediated H3K27me3, we have generated DNMT3A KO on top of NSD2dTAG/KO and have used two different PRC2 inhibitors to dissect the dependency of NSD2 function on such secondary modifications. To systematically dissect the dependency on NSD2 isoforms and its methyltransferase activity, we have reconstituted NSD2 in both NSD2 dTAG and NSD2 KO KMS-11 cells, including WT NSD2I WT, NSD2II WT, NSD2 Y1179A (LOF), NSD2 E1099K (GOF). To maximally match the timing of NSD2 perturbation, NSD2 reconstitution was generated in NSD2 dTAG and NSD2 KO cells in a Tet-ON and Tet-OFF expression system, respectively. Further we have generated KMS-11 cells overexpressing Dox-inducible H3.3K36M to investigate the consequences of abrogation of H3K36me2 by this dominant negative mutant of H3.3. We have done SLAM-seq to assess the transcriptional targets of NSD2 and will extend this with ATAC-seq and CUT&Tag to identify the molecular targets of NSD2. RESULTS: We show that NSD2 is rapidly degraded within 1 h when a degrader is added and subsequently a significant reduction of H3K36me2 is seen by 24 hours although it takes more than 3 days to see its deep depletion. We observed a reciprocal increase of H3K27me3 as previously noted. Depletion of H3K36me2 and increase in H3K27me3 was also seen in the NSD2 KO KMS-11 cells. Paradoxically we didn't see an induction of apoptosis in response to the degradation of NSD2 or its KO. In sharp contrast to the previously published shRNA results only a mild inhibition of cell proliferation was observed in NSD2 KO cells and over an extended period of 3 weeks in the dTAG system. We didn't observe an effect of H3.3 K36M overexpression on cell proliferation although it abolished H3K36me2 very efficiently. These experiments demonstrate that NSD2 is not essential for viability of KMS-11 cells but is required for optimal cell growth, which is likely independent of its H3K36me2 methyltransferase activity. SLAMseq results show that NSD2 degradation in the dTAG system causes transcriptional changes of a specific set of newly synthesized mRNAs. ATAC-seq and CUT&Tag experiments are being run and will be superimposed on the SLAM-seq data to identify immediate targets of NSD2. Subsequently, use of PRC2 inhibitors and DNMT3A KO cells in the dTAG cells and the NSD2 reconstitution experiments will uncover the mechanisms of how NSD2 controls the transcription of its targets. CONCLUSION: We show that NSD2 is not essential for viability of KMS-11 cells but is required for optimal cell growth and that NSD2 controls the transcription of a specific set of genes and does not exert a global impact on the transcriptome. We will further define the immediate targets of NSD2 by leveraging our SLAM-seq, ATAC-seq, and CUT&Tag data in NSD2 dTAG/KO and NSD2-reconstituted cells and by blocking the crosstalk with other epigenetic marks (i.e. H3K27me3 and DNAme).

Role of Circulating Tumor DNA As a Prognostic Biomarker in Patients with Diffuse Large B-Cell Lymphoma

Introduction: Nearly 50-60% of the patients withDiffuse large B-cell lymphoma (DLBCL) can be cured by front line chemoimmunotherapy. Current risk stratification modalities include cell-of-origin (germinal center (GCB) or non-GCB (activated B-cell [ABC])) subtyping, use of the revised International Prognostic Index (R-IPI), Ki-67 proliferation index assessment, and cytogenetic aberration detection. However, a novel approach for molecular residual disease (MRD) detection and treatment monitoring is needed to improve patient outcomes. Methods: A personalized and tumor-informed multiplex PCR assay (Signatera™ bespoke mPCR NGS assay) was used for the detection and quantification of ctDNA in a prospective clinical cohort of patients with DLBCL. Serial ctDNA testing was performed at various time points collected at the treating physician's discretion, including pretreatment (baseline), during first-line therapy, during and post-salvage therapy, at the end of treatment, and during surveillance, up to the last clinical follow-up. ctDNA clearance was defined as change in ctDNA status (positive to negative) from the pre-treatment time point to the subsequent time point. Event-free survival (EFS) was defined as the time elapsed since the initial ctDNA clearance or PET imaging, for those without ctDNA clearance, during which the patient did not experience progression, death or treatment with salvage therapy. EFS was censored at the last date of followup if the patient was alive and without an event. Overall survival (OS) was defined as the elapsed time after primary treatment until death from any cause. Univariate and multivariable models were analyzed using Cox proportional models. Results: In this retrospective evaluation of commercial ctDNA testing in patients with stage I-IV DLBCL, a total of 50 patients (I:3, II:12, III: 4, and IV: 29;median age: 59 years) were analyzed. Patients were followed up for a median of 12.68 months (inter quartile range: 7.9-17.7 months). Of the 50 patients, 41 had pre-treatment (baseline) time points available and ctDNA was detected in 95% (39/41) of patients. Since the majority of the patients had stage IV disease, median ctDNA detection levels were observed to be high (overall median: 534 MTM/ml, range: 0-5957 MTM/ml; I median: 9.6 MTM/mL [range: 0.2-2822 MTM/mL], II median: 53 MTM/mL [range: 0.8-862 MTM/mL], III median: 368 [range: 2-940 MTM/mL], IV median: 1011 MTM/mL [range: 74-5958 MTM/mL]). Clearance of ctDNA at any point during first-line therapy was associated with improved outcomes (EFS, HR: 8.3, 95% CI: 2.56-27.0, p<0.0001 and OS, HR: 15.2, 95% CI: 1.53-149, p=0.0021), compared to patients who remained ctDNA-positive. Furthermore, 48% (13/27) of patients were found to clear their ctDNA upon the completion of the first cycle of treatment. On comparing baseline ctDNA levels with other clinicopathological risk factors, high ctDNA levels were observed to be strongly correlated with higher R-IPI scores (0-2 vs. 3-5; p=0.0008) and higher stage (I-II vs. III-IV; p=0.0007). Additionally, multivariate analysis determined that ctDNA clearance significantly outperformed all other clinicopathologic risk factors associated with EFS (HR: 48.8, 95% CI:1.2-2005, p=0.04). Finally, ctDNA clearance predicted complete response (CR)/no evidence of disease (NED) on average 95 days (range: 0-14.7 months) ahead of PET imaging or biopsy. Conclusions: In this study utilizing a real-world cohort of patients with stage I-IV DLBCL, ctDNA clearance during first-line therapy was prognostic for EFS and OS and preceded clinical response. ctDNA testing in patients with DLBCL may enable personalized surveillance, intervention, and/or trial options, ultimately improving patient outcomes.

The value of serum creatinine as biomarker of disease progression in spinal and bulbar muscular atrophy (SBMA)

Serum creatinine has been indicated as a potential marker of motor function in SBMA and results form previous longitudinal studies pointed to its decline over time. This is a longitudinal retrospective study investigating creatinine changes over a 36-month-period in 73 patients with SBMA. Severity and progression of the disease was assessed according to serum creatine kinase (CK) values, manual muscle testing (MMT), SBMA functional rating scale (SBMAFRS) score, 6-min-walk test (6MWT) value, and spirometry (forced vital capacity, fVC%) obtained at the baseline and at each of the annual follow-up visits. Baseline serum creatinine concentrations positively correlated with 6MWT, the MMT megascore score of both the upper (ULM) and lower (LLM) limbs and SBMAFRS. No correlation was found with CK or fVC% values. Similar correlation results were achieved at all the subsequent time points. Longitudinal assessments conducted by the generalized estimating equations (GEE) method returned significant changes for SBMAFRS (− 1.41 points per year, p < 0.001), ULM and LLM (− 0.69, p = 0.01; and − 1.07, p < 0.001, respectively), 6MWT (− 47 m, p < 0.001) but not for creatinine (− 0.82, p > 0.05). We also observed that creatinine levels at baseline did not correlate with changes in the other measures from baseline at each annual visit. Our data do not support a role for serum creatinine as sensitive biomarker of disease progression, and possibily prognosis, in SBMA.

Respiratory mechanics and mechanical power during low vs. high positive end-expiratory pressure in obese surgical patients – A sub-study of the PROBESE randomized controlled trial

Study objectiveWe aimed to characterize intra-operative mechanical ventilation with low or high positive end-expiratory pressure (PEEP) and recruitment manoeuvres (RM) regarding intra-tidal recruitment/derecruitment and overdistension using non-linear respiratory mechanics, and mechanical power in obese surgical patients enrolled in the PROBESE trial. DesignProspective, two-centre substudy of the international, multicentre, two-arm, randomized-controlled PROBESE trial. SettingOperating rooms of two European University Hospitals. PatientsForty-eight adult obese patients undergoing abdominal surgery. InterventionsIntra-operative protective ventilation with either PEEP of 12 cmH2O and repeated RM (HighPEEP+RM) or 4 cmH2O without RM (LowPEEP). MeasurementsThe index of intra-tidal recruitment/de-recruitment and overdistension (%E2) as well as airway pressure, tidal volume (VT), respiratory rate (RR), resistance, elastance, and mechanical power (MP) were calculated from respiratory signals recorded after anesthesia induction, 1 h thereafter, and end of surgery (EOS). Main resultsTwenty-four patients were analyzed in each group. PEEP was higher (mean ± SD, 11.7 ± 0.4 vs. 3.7 ± 0.6 cmH2O, P < 0.001) and driving pressure lower (12.8 ± 3.5 vs. 21.7 ± 6.8 cmH2O, P < 0.001) during HighPEEP+RM than LowPEEP, while VT and RR did not differ significantly (7.3 ± 0.6 vs. 7.4 ± 0.8 ml∙kg−1, P = 0.835; and 14.6 ± 2.5 vs. 15.7 ± 2.0 min−1, P = 0.150, respectively). %E2 was higher in HighPEEP+RM than in LowPEEP following induction (−3.1 ± 7.2 vs. -12.4 ± 10.2%; P < 0.001) and subsequent timepoints. Total resistance and elastance (13.3 ± 3.8 vs. 17.7 ± 6.8 cmH2O∙l∙s−2, P = 0.009; and 15.7 ± 5.5 vs. 28.5 ± 8.4 cmH2O∙l, P < 0.001, respectively) were lower during HighPEEP+RM than LowPEEP. Additionally, MP was lower in HighPEEP+RM than LowPEEP group (5.0 ± 2.2 vs. 10.4 ± 4.7 J∙min−1, P < 0.001). ConclusionsIn this sub-cohort of PROBESE, intra-operative ventilation with high PEEP and RM reduced intra-tidal recruitment/de-recruitment as well as driving pressure, elastance, resistance, and mechanical power, as compared with low PEEP. Trial registrationThe PROBESE study was registered at www.clinicaltrials.gov, identifier: NCT02148692 (submission for registration on May 23, 2014).

Comparison of Outcomes After Reverse Total Shoulder Arthroplasty in Patients With Proximal Humerus Fractures Versus Rotator Cuff Arthropathy.

Indications for reverse total shoulder arthroplasty (rTSA) has expanded to encompass complex proximal humerus fractures (PHFs) in recent years. The purpose of this study was to report and assess whether PHF patients treated with rTSA could achieve similar functional outcomes and short-term survivorship to patients who underwent rTSA for rotator cuff arthropathy (RTCA). All consecutive patients with a preoperative diagnosis of PHF or RTCA, 18 years or older, treated with rTSA at a single academic institution between 2018 and 2020 with a minimum 2-year follow-up were retrospectively reviewed. Primary outcomes were survivorship defined as revision surgery or implant failure analyzed using the Kaplan-Meier survival curve, and functional outcomes, which included Quick Disabilities of the Arm, Shoulder, and Hand, and range of motion (ROM) were compared at multiple follow-up time points up to 2 years. Secondary outcomes were patient demographics, comorbidities, surgical data, length of hospital stay, and discharge disposition. A total of 48 patients were included: 21 patients (44%) were diagnosed with PHF and 27 patients (56%) had RTCA. The Kaplan-Meier survival rate estimates at 3 years were 90.5% in the PHF group and 85.2% in the RTCA group. No differences in revision surgery rates between the two groups (P = 0.68) or survivorship (P = 0.63) were found. ROM was significantly lower at subsequent follow-up time points in multiple planes (P < 0.05). A greater proportion of patients in the PHF group received cement for humeral implant fixation compared with the RTCA group (48% versus 7%, P = 0.002). The mean length of hospital stay was longer in PHF patients compared with RTCA patients (2.9 ± 3.8 days versus 1.6 ± 1.8 days, P = 0.13), and a significantly lower proportion of PHF patients were discharged home (67% versus 96%, P = 0.015). The rTSA implant survivorship at 3 years for both PHF and RTCA patients show comparable results. At the 2-year follow-up, RTCA patients treated with rTSA were found to have better ROM compared with PHF patients.

Role of Insole Material in Treatment of Plantar Fasciitis: A Randomized Clinical Trial

Category: Hindfoot; Other Introduction/Purpose: Plantar fasciitis (PF), a leading cause of persistent heel pain, results in almost a million physician visits annually. Conservative treatment is often the first line of management with insoles being frequently prescribed. While multiple studies have compared insoles based on the degree of customization to foot contour, the literature is lacking in data comparing insoles based on their material. In this randomized clinical trial, we compared the early effects of foam, polyurethane, and carbon fiber insoles in the treatment of PF, using a robust set of PROMS. Methods: A randomized clinical trial was designed at the foot and ankle research center of a tertiary care hospital in Massachusetts. Adult patients diagnosed with PF who had not received injectable or surgical treatment for it were included. Participants who consented were randomly allocated one of the three prefabricated insoles – carbon fiber insole (Group 1, n=13), polyurethane insole (Group 2, n=13), or foam insole (Group 3, n=9) for regular use. Their response was recorded using PROMIS 3a (for pain intensity), PROMIS 4a (for pain interference), FAOS (Foot and Ankle Outcome Score), and VAS for pain at baseline (T0), two (T2), six (T6) and twelve weeks (T12). A per-proto analysis was undertaken using Kruskal Wallis and Friedman tests. P&lt; 0.05 was set as the level of significance. Results: Among groups 1, 2 and 3, the mean age of participants was 51.9±13.2, 51±12.5, and 47.9±13.5 years respectively and baseline weight varied as 211.6±32.7 lbs, 193.6±59.3 lbs, and 180.7±33 lbs respectively (p&gt;0.05 for both). The group-wise distribution of participants based on sex and laterality of the affected foot showed no difference. Participants in each group reported similar intensity of the pain (VAS) at baseline and at all follow-up time points (p&gt;0.05). A comparison of outcomes between baseline and subsequent timepoints showed significant improvement in quality of life (FAOS) at T12 (p=0.047) for carbon fibre insole users (Figure 1), significant pain relief on PROMIS Pain intensity and Pain Interference questionnaires for foam insoles at T3 and T12 (p=0.0003, 0.036 respectively). Conclusion: Carbon fiber insoles offered slightly better pain relief as early as six weeks into treatment and improved quality of life after twelve weeks of use. A possible link between the material of insole used and effective treatment of PF may exist. Conducting further analyses on the position of the foot during gait while using insoles and designing the insoles based on patient-specific criteria should be considered for future research.

Medications for opioid use disorder prescribed at hospital discharge associated with decreased opioid agonist dispensing in patients with opioid use disorder requiring critical care: A retrospective study

IntroductionBuprenorphine is highly effective for the treatment of opioid use disorder (OUD), and, in recent years, the rates of patients maintained on buprenorphine requiring critical care have been steadily increasing. Currently, no unified guidance exists for buprenorphine management during critical illness. Likewise, we do not know if patients maintained on buprenorphine for OUD are prescribed medications for OUD (MOUD) following hospital discharge or if buprenorphine management influences mu opioid agonist dispensing. MethodsIn our cohort of adults over the age of 18 with OUD, receiving buprenorphine formulations in the 3 months preceding their ICU admission, we sought to investigate the relationship between receipt of MOUD and non-MOUD opioid prescribing up to 12 months following hospital discharge. This was a single-center, retrospective cohort study approved by the MaineHealth institutional review board. The study analyzed differences in prescription rates between discharge and subsequent time points using chi square or Fisher's exact test, as appropriate. We performed analyses using SPSS Statistical Software version 28 (IBM SPSS Inc., Armonk, NY) with significance set at p < 0.05. ResultsWe identified a statistically significant increase in MOUD prescribing 3 months posthospital discharge in patients who received MOUD at time of discharge (87.9 % vs 40 % p = 0.002.) The study found a significant increase in nonbuprenorphine opioid prescribing in patients who did not receive an MOUD prescription at time of discharge (24.2 % vs 70 % p = 0.007). This trend persisted at the 6-month and 12-month time points; however, it did not reach statistical significance. Additionally, the study identified a significant reduction in the incidence of non-MOUD opioid dispensing in patients prescribed MOUD at each time point measured (p = 0.007, p < 0.001. p < 0.001 and p = 0.008 at discharge, 3, 6, and 12 months, respectively). ConclusionsThese findings support continuing buprenorphine dispensing following hospital discharge.

Docosahexaenoic Acid Supplementation in Lactating Women Increases Breast Milk and Erythrocyte Membrane Docosahexaenoic Acid Concentrations and Alters Infant n–6:n–3 Fatty Acid Ratio

BackgroundLow concentrations of docosahexaenoic acid (DHA) or high n–6 (ω-6):n–3 ratio in pregnant women is associated with poor fetal growth velocity and suboptimal neurodevelopment. However, there is a lack of data on levels of important n–6 and n–3 fatty acids (FAs) at different time points during pregnancy and lactation from India. Data on how much DHA is transferred during actual supplementation are also scarce. ObjectivesWe report the concentrations of n–6 and n–3 FAs in maternal and infant blood and in breast milk following maternal supplementation with DHA or placebo. MethodsA total of 957 pregnant women (≤20 wk) from Belagavi, Karnataka, were randomly assigned to receive either 400 mg/d of algal DHA or placebo through 6 mo postpartum. Blood samples were collected from the mother at recruitment/baseline, delivery, and 6 mo postpartum and from the infant at birth (cord) and 12 mo (venous). Breast milk samples were collected from a subsample at delivery, 1 mo and 6 mo postpartum. The FA profile was analyzed using gas chromatography. ResultsThe concentration of DHA appeared to be higher in erythrocyte and breast milk samples of the DHA-supplemented group at all subsequent time points. The n–6:n–3 ratio was lower among women in the DHA group at delivery [DHA: 4.08 (1.79); placebo: 5.84 (3.57); P < 0.001] and at 6 mo postpartum [DHA: 5.34 (2.64); placebo: 7.69 (2.9); P < 0.001]. Infants of DHA-supplemented mothers also had a lower n–6:n–3 ratio at delivery and 12 mo. The n–6:n–3 ratio of breast milk increased from delivery through 1 to 6 mo but remained lower in the DHA-supplemented group than in the placebo. ConclusionsMaternal DHA supplementation with 400 mg/d from early pregnancy through 6 mo postpartum significantly increased circulating DHA in breast milk and infant erythrocyte, whereas decreased erythrocyte and breast milk n–6:n–3 ratio. However, maternal supplementation did not get the ratio to the recommended levels.

An Analysis of Optical Coherence Tomography Angiography (OCT-A) Perfusion Density Maps in Patients Treated for Retinal Vein Occlusion with Intravitreal Aflibercept.

The primary goal of this study was to evaluate the reduction in non-perfusion area and improvement in blood flow as well as the reduction in retinal edema on optical coherence tomography angiography (OCT-A) in patients with retinal vein occlusion treated with 2 mg intravitreal injections of aflibercept. Fifty eyes of nine patients with central retinal vein occlusion (CRVO) and sixteen patients with branch retinal vein occlusion (BRVO), aged 50 to 75 years, were collectively analyzed as retinal vein occlusion (RVO). The following parameters were analyzed: superficial vessel density (VDSF), deep vessel density (VDD), flow area in the outer retina (FAOR), choriocapillaris flow area (FACC), non-flow area (NFA) and the foveal avascular zone (FAZ). OCT-A revealed a reduction in macular edema. The most significant change in central retinal thickness (CRT) was observed between measurement timepoint "5" and the baseline (46%). The non-flow area was also reduced. Following a significant decrease in superficial vessel density 30 days after the first dose of aflibercept, a non-significant increase was noted at the subsequent timepoints. An increase was also found in deep vessel density and choriocapillaris flow area. Improvements in the above OCT-A parameters resulted in increased retinal blood flow and improved visual acuity. Patients with retinal vein occlusion treated with 2 mg aflibercept exhibited reduced macular edema and FAZ, increased vessel density, improved blood flow, and better visual acuity.

Liver Glycogen as a Sensitive Indicator of Food Limitation in Delta Smelt

AbstractAssessing habitat quality is a major goal of conservationists and restoration practitioners, but to associate habitat quality with biomarkers of vagile animals, the biomarkers must respond rapidly. Here, we identified a biomarker capable of rapidly detecting food limitation in the imperiled Delta Smelt (Hypomesus transpacificus), a pelagic fish endemic to the San Francisco Estuary (SFE). We conducted an experiment with fed and unfed treatments of hatchery-raised, sub-adult Delta Smelt that were sampled at 12 time points: 0, 1, 2, 3, 4, 5, 6, 7, 9, 11, 14, and 21 days. We then compared four biomarkers using Day 21 fish: RNA/DNA in liver, triglycerides in liver, glycogen in liver, and glycogen in muscle. Of the liver endpoints, glycogen had the largest difference between treatments at Day 21, so we compared it to muscle glycogen across all time points. Liver glycogen declined by 60% after 1 day of fasting and remained depressed in the fasting treatment across all subsequent time points. Muscle glycogen also responded rapidly, taking only 2 days to decline by 39% in the fasting treatment, but the difference was inconsistent across subsequent time points. When applied to hatchery-released Delta Smelt collected from the SFE, liver glycogen concentrations were less than half that of the fed hatchery fish, consistent with the hypothesis of food limitation in the wild, but also several other potential causes. This study highlights the utility of liver glycogen as an indicator of recent foraging success in Delta Smelt.

Epicardial Adipose Tissue Changes during Statin Administration in Relation to the Body Mass Index: A Longitudinal Cardiac CT Study

The epicardial adipose tissue (EAT) is the visceral fat located between the myocardium and the pericardium. We aimed to perform a longitudinal evaluation of the epicardial adipose tissue using an advanced computer-assisted approach in a population of patients undergoing Cardiac CT (CCT) during statin administration, in relation to their body mass index (BMI). We retrospectively enrolled 95 patients [mean age 62 ± 10 years; 68 males (72%) and 27 females (28%)] undergoing CCT for suspected coronary artery disease during statin administration. CCT was performed at two subsequent time points. At the second CCT, EAT showed a mean density increase (−75.59 ± 7.0 HU vs. −78.18 ± 5.3 HU, p &lt; 0.001) and a volume decrease (130 ± 54.3 cm3 vs.142.79 ± 56.9 cm3, p &lt; 0.001). Concerning coronary artery EAT thickness, a reduction was found at the origin of the right coronary artery (13.26 ± 5.2 mm vs. 14.94 ± 5.8, p = 0.001) and interventricular artery (8.22 ± 3.7 mm vs. 9.13 ± 3.9 mm, p = 0.001). The quartile (Q) attenuation percentage (%) distribution of EAT changed at the second CCT. The EAT % distribution changed by the BMI in Q1 (p = 0.015), Q3 (p = 0.001) and Q4 (p = 0.043) at the second CCT, but the normal-BMI and overweight/obese patients showed a similar response to statin therapy in terms of quartile distribution changes. In conclusion, statins may determine significant changes in EAT quantitative and qualitative characteristics detected by CCT; the BMI influences the EAT composition, but statins determine a similar response in quartile distribution’s variation, irrespective of the BMI.

Is foam rolling as effective as its popularity suggests? A randomised crossover study exploring post-match recovery in female basketball

ABSTRACT This study aimed to assess the effectiveness of foam rolling as a post-match recovery tool in basketball. Using a crossover design, 13 female players completed two simulated matches, followed by foam rolling (FR) or placebo (CON). Countermovement jump height (CMJ), heart rate variability (Ln-rMSSD), muscle soreness (VAS), perceived recovery (TQR) and fatigue (Rating-of-Fatigue) were recorded at pre-match, post-match, post-recovery and 24 h post-match. No significant effect of time*intervention and intervention were found for any variable (p > 0.05), while a significant effect of time (p < 0.01) was reported for all variables. Post-hoc analyses revealed lower CMJ and Ln-rMSSD at post-match compared with all other time points (p < 0.001), increases in pre-match VAS scores at all subsequent time points (p < 0.01), and worse TQR and Rating-of-Fatigue scores from pre-to-post-match and pre-match-to-post-recovery (p < 0.01), except for unchanged TQR values from pre-match-to-post-recovery in FR (p > 0.05). Overall, the present data suggest that foam rolling was generally ineffective for improving post-exercise recovery in female basketball players after a single match. Future research investigating the effectiveness of foam rolling in players of different sex, age and/or competitive level is warranted to provide further insight on the topic.

Abstract PO-045: Utility of circulating tumor tissue modified viral (TTMV)-HPV DNA to resolve clinically indeterminate findings in patients following definitive treatment for HPV-driven oropharyngeal cancer

Abstract Introduction: Indeterminate results of clinical or imaging examinations are common across the care continuum, and decisions made can lead to overtreatment and psychological and financial harms for the patient. Additionally, due to the emergence of highly sensitive circulating tumor DNA assays, an indeterminate clinical status can also arise when molecular detection precedes clinical or radiological evidence of recurrence for a period of time. We recently reported &amp;gt;99% negative and &amp;gt;95% positive predictive value of circulating tumor tissue modified viral (TTMV)-HPV DNA for recurrence detection in a large, real-world cohort of patients with HPV-driven oropharyngeal cancer. Here we quantified the frequency of clinically indeterminate findings (CIF) in this patient cohort and assessed the prognostic value of TTMV-HPV DNA to resolve these CIF. Methods: This IRB-approved, retrospective observational cohort study included 543 patients across eight U.S. centers who were ≥3 months post-treatment for HPV-driven OPSCC. Patients had one or more TTMV-HPV DNA results (NavDx®, Naveris Laboratories) obtained during surveillance between February 2020 and January 2022 (n=1372 total tests), with mean follow-up time of 24 months (range: 4.9-185). Test results were correlated with physician-reported exam and imaging findings to assess disease status in follow-up. Results: Of the 543 patients, 245 patients (45.1%) had 797 CIF by imaging and/or clinical examination during their surveillance period. Of these 245 patients, 71 patients had 93 CIF that had a coincident TTMV-HPV DNA result (either negative or positive) available within 3 months of the CIF. The TTMV-HPV DNA result was able to correctly predict the presence or absence of recurrence across 91 of the 93 (97.8%) individual CIF. Both incorrectly predicted CIF were in the setting of an undetectable TTMV-HPV DNA result. One patient had their recurrence detected by chest CT 63 days later and had a positive TTMV-HPV DNA test the same day. The second patient had a positive TTMV-HPV DNA test during their subsequent immunotherapy. In contrast to the 45% of patients with CIF as a result of imaging and/or clinical examination, only 18 patients (3.3%) had CIF due to a positive TTMV-HPV DNA result that was not accompanied by clinical evidence of recurrence within 3 months. Importantly, of these 18 patients, 15 patients had confirmed recurrence at a subsequent time point (median time to recurrence: 232 days), leaving only 3 of 543 (0.6%) patients with an ongoing indeterminate recurrence status. Conclusion: Clinical indeterminate findings as a result of imaging and/or clinical examination are common, whereas clinically indeterminate status due to circulating TTMV-HPV DNA testing is rare. Our findings strongly support the clinical utility and value of monitoring circulating TTMV-HPV DNA during post-treatment surveillance to inform interpretation of indeterminate clinical findings. Citation Format: Scott Roof, Glenn Hanna, James Jabalee, Eleni Rettig, Rocco Ferrandino, Sida Chen, Marshall R. Posner, Krzysztof J. Misiukiewicz, Eric M. Genden, Raymond L. Chai, John Sims, Elaine Thrash, Scott Stern, Noah S. Kalman, Sreenija Yarlagadda, Adam Raben, Lydia Clements, Abie Mendelsohn, John M. Kaczmar, Yadav Pandey, Mihir Bhayani, Piyush Gupta, Charlotte Kuperwasser, Catherine Del Vecchio Fitz, Barry M. Berger. Utility of circulating tumor tissue modified viral (TTMV)-HPV DNA to resolve clinically indeterminate findings in patients following definitive treatment for HPV-driven oropharyngeal cancer [abstract]. In: Proceedings of the AACR-AHNS Head and Neck Cancer Conference: Innovating through Basic, Clinical, and Translational Research; 2023 Jul 7-8; Montreal, QC, Canada. Philadelphia (PA): AACR; Clin Cancer Res 2023;29(18_Suppl):Abstract nr PO-045.

Variation of corticospinal excitability during kinesthetic illusion induced by musculotendinous vibration.

Despite being studied for more than 50 years, the neurophysiological mechanisms underlying vibration (VIB)-induced kinesthetic illusions are still unclear. The aim of this study was to investigate how corticospinal excitability tested by transcranial magnetic stimulation (TMS) is modulated during VIB-induced illusions. Twenty healthy adults received vibration over wrist flexor muscles (80 Hz, 1 mm, 10 s). TMS was applied over the primary motor cortex representation of wrist extensors at 120% of resting motor threshold in four random conditions (10 trials/condition): baseline (without VIB), 1 s, 5 s, and 10 s after VIB onset. Means of motor-evoked potential (MEP) amplitudes and latencies were calculated. Statistical analysis found a significant effect of conditions (stimulation timings) on MEP amplitudes (P = 0.035). Paired-comparisons demonstrated lower corticospinal excitability during VIB at 1 s compared with 5 s (P = 0.025) and 10 s (P = 0.003), although none of them differed from baseline values. Results suggest a time-specific modulation of corticospinal excitability in muscles antagonistic to those vibrated, i.e., muscles involved in the perceived movement. An early decrease of excitability was observed at 1 s followed by a stabilization of values near baseline at subsequent time points. At 1 s, the illusion is not yet perceived or not strong enough to upregulate corticospinal networks coherent with the proprioceptive input. Spinal mechanisms, such as reciprocal inhibition, could also contribute to lower the corticospinal drive of nonvibrated muscles in short period before the illusion emerges. Our results suggest that neuromodulatory effects of VIB are likely time-dependent, and that future work is needed to further investigate underlying mechanisms.NEW & NOTEWORTHY The modulation of corticospinal excitability when perceiving a vibration (VIB)-induced kinesthetic illusion evolves dynamically over time. This modulation might be linked to the delayed occurrence and progressive increase in strength of the illusory perception in the first seconds after VIB start. Different spinal/cortical mechanisms could be at play during VIB, depending on the tested muscle, presence/absence of an illusion, and the specific timing at which corticospinal drive is tested pre/post VIB.

Examining the effects of pre-competition rapid weight loss on hydration status and competition performance in elite judo athletes

The prevalence of rapid weight loss (RWL) among martial arts athletes including judo is very high. Many applied RWL strategies could be dangerous to health and even lead to death. Therefore, the International Judo Federation (IJF) introduced changes in the weigh-in rules, changing the official weigh-in for the day before the competition. Thus, the purpose of this study was to examine the impact of the new IJF rules on hydration status and weight loss strategies among professional judo athletes. Seventeen elite judo athletes participated in the study. Body mass and hydration status, were analyzed before the competition. Moreover, competition result and practice of RWL survey were collected. All subjects reached their weight category limits for the competition. RWL resulted in body mass changes (p < 0.001, ηp2 = 0.79) and dehydration among participants (urine osmolality > 700 [mOsmol*kg]−1 and urine specific gravity > 1.020 [g*cm3]−1). However, urine osmolality (p > 0.05, ηp2 = 0.18), as well as urine specific gravity (p > 0.05, ηp2 = 0.16), at subsequent time points of measurement revealed no statistical differences. The prevalence of RWL was 100%, and only 17.6% of the athletes declared that they would compete in a different weight category if the competition would be conducted on the same day of the weigh-in. All judo athletes applied RWL procedures using traditional methods to achieve the required body mass (i.e., increased exercise, reduced fluid, and food intake). Dehydration state was not associated with competitive performance (p > 0.05).

Efficacy of online communication partner training package for student healthcare professionals.

People with aphasia are vulnerable recipients of healthcare. The nature of the communicative environment and the communication disability can adversely impact access to timely and quality healthcare. Student healthcare professionals are often underprepared to interact successfully with people with aphasia and may benefit from communication partner training (CPT). To investigate the potential effectiveness and acceptability of a brief, two-part introductory Supported Conversation for Adults with Aphasia (SCA™)-based CPT package, delivered to a sample of students across a diverse range of healthcare disciplines. A pre-post-within group experimental design was used to investigate the potential effectiveness and acceptability of an online CPT package (50 minute module + 1 hour workshop) for healthcare students. The Aphasia Attitudes, Strategies and Knowledge (AASK) survey measured participants' knowledge of aphasia, facilitative communication strategies and attitudes towards people with aphasia. Data were collected pre-training, following the training module and following the workshop, and 6 weeks post-training. Statistical analysis was conducted on the AASK data. In addition, participant feedback (ratings and open text responses) was collected after the workshop. Ratings were analysed descriptively, and thematic content analysis was used for open text responses. 236 participants completed the pre-training AASK and 106 completed the AASK at subsequent time points. Statistically significant gains were demonstrated from pre- to post-module completion. Between the end of the module and the end of the workshop, some gains were maintained and others showed further statistically significantly improvements. While all gains were not maintained at the 6-week follow-up, statistically significantly improvements from pre-training scores remained evident. Student feedback was predominantly positive, with suggested improvements for training content and length. The results provide preliminary evidence that a brief, online CPT package can support student healthcare professionals' knowledge and attitudes towards aphasia and communicating with people with aphasia. Online training was acceptable to students and feasible as an embedded or optional component of curriculum. Ongoing training (e.g., in the form of refresher sessions) and inclusion of a skills-based component are recommended to maximize communication skill development. What is already known on the subject Student healthcare professionals recognize the need to develop knowledge and skills to successfully support people with communication disability, such as aphasia, to participate effectively in their healthcare. Evidence in favour of online communication partner training for student healthcare professionals is currently limited. What this study adds to the existing knowledge This study demonstrates that a brief introductory online communication partner training program can be efficacious for improving knowledge and attitudes regarding communicating with people who have aphasia. What are the potential or actual clinical implications of this work? Students will likely need further ongoing refresher training with inclusion of practical components to develop and maintain the knowledge and skills required to be proficient communication partners with people with aphasia.

MR-based spatiotemporal anisotropic atrophy evaluation of hippocampus in Alzheimer's disease progression by multiscale skeletal representation.

Increasing evidence has shown a higher sensitivity of Alzheimer's disease (AD) progression by local hippocampal atrophy rather than the whole volume. However, existing morphological methods based on subfield-volume or surface in imaging studies are not capable to describe the comprehensive process of hippocampal atrophy as sensitive as histological findings. To map histological distinctive measurements onto medical magnetic resonance (MR) images, we propose a multiscale skeletal representation (m-s-rep) to quantify focal hippocampal atrophy during AD progression in longitudinal cohorts from the Alzheimer's Disease Neuroimaging Initiative (ADNI). The m-s-rep captures large-to-small-scale hippocampal morphology by spoke interpolation over label projection on skeletal models. To enhance morphological correspondence within subjects, we align the longitudinal m-s-reps by surface-based transformations from baseline to subsequent timepoints. Cross-sectional and longitudinal measurements derived from m-s-rep are statistically analyzed to comprehensively evaluate the bilateral hippocampal atrophy. Our findings reveal that during the early AD progression, atrophy primarily affects the lateral-medial extent of the hippocampus, with a difference of 1.8 mm in lateral-medial width in 2 years preceding conversion (p < .001), and the medial head exhibits a maximum difference of 3.05%/year in local atrophy rate (p = .011) compared to controls. Moreover, progressive mild cognitive impairment (pMCI) exhibits more severe and widespread atrophy in the head and body compared to stable mild cognitive impairment (sMCI), with a maximum difference of 1.21 mm in thickness in the medial head 1 year preceding conversion (p = .012). In summary, our proposed method can quantitatively measure the hippocampal morphological changes on 3T MR images, potentially assisting the pre-diagnosis and prognosis of AD.

Whole Blood DNA Methylation Changes Are Associated with Anti-TNF Drug Concentration in Patients with Crohn's Disease.

Anti-tumour necrosis factor [TNF] treatment failure in patients with inflammatory bowel disease [IBD] is common and frequently related to low drug concentrations. In order to identify patients who may benefit from dose optimisation at the outset of anti-TNF therapy, we sought to define epigenetic biomarkers in whole blood at baseline associated with anti-TNF drug concentrations at week 14. DNA methylation from 1104 whole blood samples from 385 patients in the Personalised Anti-TNF Therapy in Crohn's disease [PANTS] study were assessed using the Illumina EPIC Beadchip [v1.0] at baseline and weeks 14, 30, and 54. We compared DNA methylation profiles in anti-TNF-treated patients who experienced primary non-response at week 14 if they were assessed at subsequent time points and were not in remission at week 30 or 54 [infliximab n = 99, adalimumab n = 94], with patients who responded at week 14 and when assessed at subsequent time points were in remission at week 30 or 54 [infliximab n = 99, adalimumab n = 93]. Overall, between baseline and week 14, we observed 4999 differentially methylated positions [DMPs] annotated to 2376 genes following anti-TNF treatment. Pathway analysis identified 108 significant gene ontology terms enriched in biological processes related to immune system processes and responses. Epigenome-wide association [EWAS] analysis identified 323 DMPs annotated to 210 genes at baseline associated with higher anti-TNF drug concentrations at Week 14. Of these, 125 DMPs demonstrated shared associations with other common traits [proportion of shared CpGs compared with DMPs] including body mass index [23.2%], followed by C-reactive protein [CRP] [11.5%], smoking [7.4%], alcohol consumption per day [7.1%], and IBD type [6.8%]. EWAS of primary non-response to anti-TNF identified 20 DMPs that were associated with both anti-TNF drug concentration and primary non-response to anti-TNF with a strong correlation of the coefficients [Spearman's rho = -0.94, p <0.001]. Baseline DNA methylation profiles may be used as a predictor for anti-TNF drug concentration at week 14 to identify patients who may benefit from dose optimisation at the outset of anti-TNF therapy.

Exercise promotes growth and rescues volume deficits in the hippocampus after cranial radiation in young mice.

Human and animal studies suggest that exercise promotes healthy brain development and function, including promoting hippocampal growth. Childhood cancer survivors that have received cranial radiotherapy exhibit hippocampal volume deficits and are at risk of impaired cognitive function, thus they may benefit from regular exercise. While morphological changes induced by exercise have been characterized using magnetic resonance imaging (MRI) in humans and animal models, evaluation of changes across the brain through development and following cranial radiation is lacking. In this study, we used high-resolution longitudinal MRI through development to evaluate the effects of exercise in a pediatric mouse model of cranial radiation. Female mice received whole-brain radiation (7 Gy) or sham radiation (0 Gy) at an infant equivalent age (P16). One week after irradiation, mice were housed in either a regular cage or a cage equipped with a running wheel. In vivo MRI was performed prior to irradiation, and at three subsequent timepoints to evaluate the effects of radiation and exercise. We used a linear mixed-effects model to assess volumetric and cortical thickness changes. Exercise caused substantial increases in the volumes of certain brain regions, notably the hippocampus in both irradiated and nonirradiated mice. Volume increases exceeded the deficits induced by cranial irradiation. The effect of exercise and irradiation on subregional hippocampal volumes was also characterized. In addition, we characterized cortical thickness changes across development and found that it peaked between P23 and P43, depending on the region. Exercise also induced regional alterations in cortical thickness after 3 weeks of voluntary exercise, while irradiation did not substantially alter cortical thickness. Our results show that exercise has the potential to alter neuroanatomical outcomes in both irradiated and nonirradiated mice. This supports ongoing research exploring exercise as a strategy for improving neurocognitive development for children, particularly those treated with cranial radiotherapy.

Application of SLE-DAS to assess subcutaneous belimumab efficacy in a cohort of systemic lupus erythematosus patients.

To assess the efficacy of subcutaneous (sc) belimumab (BLM) by the application of SLE-DAS in a monocentric SLE cohort. We evaluated SLE patients treated with sc BLM from March 2019. Disease activity has been assessed by SLEDAI-2k, SLE-DAS and PGA (Physician Global Assessment) in all the established time-points [baseline (T0), after 1 (T1), 3 (T3), 6 (T6) and 12 (T12) months]. Furthermore, we applied and compared the achievement of remission according to SLE-DAS values (SLEDAS ≤2.08 + PDN ≤5mg/daily) and DORIS definition (clinical SLEDAI- 2k=0 + PGA<0.5 + antimalarial treatment, PDN≤5mg/daily, stable immunosuppressive treatment). We enrolled 86 patients [M/F 5/81, median age 48 years (IQR 17.5), median disease duration 166 months (IQR 216)]. At baseline, median values of SLEDA-2k and SLE-DAS were 6 (IQR 4) and 5.77 (IQR 4.33), respectively, and they significantly correlated (r=0.719, CI 95% 0.586-0.815, p<0.0001). Median duration of treatment was 14 months (IQR 20). We found a significant reduction of SLEDAI-2k and SLE-DAS already at T1, maintained in the subsequent time-points (p<0.0001). At T12, a remission state was achieved by 60.4% of patients according to SLE-DAS definition and by 62.3% according to the DORIS definition. Both definitions of remission have demonstrated an agreement of 84%, with a Cohen's kappa equal to 0.6. In this study we applied SLE-DAS to assess the efficacy of sc BLM, by analysing its over-time changes and by comparing its performance with SLEDAI-2k. Indeed, our results suggest the usefulness of this new activity index in a real-life setting.