Stealth transgenes enable CAR-T cells to evade host immune responses.

BACKGROUND: Adoptive cell therapy, such as chimeric antigen receptor (CAR)-T cell therapy, has improved patient outcomes for hematological malignancies. Currently, four of the six FDA-approved CAR-T cell products use the FMC63-based αCD19 single-chain variable fragment, derived from a murine monoclonal antibody, as the extracellular binding domain. Clinical studies demonstrate that patients develop humoral and cellular immune responses to the non-self CAR components of autologous CAR-T cells or donor-specific antigens of allogeneic CAR-T cells, which is thought to potentially limit CAR-T cell persistence and the success of repeated dosing. METHODS: In this study, we implemented a one-shot approach to prevent rejection of engineered T cells by simultaneously reducing antigen presentation and the surface expression of both Classes of the major histocompatibility complex (MHC) via expression of the viral inhibitors of transporter associated with antigen processing (TAPi) in combination with a transgene coding for shRNA targeting class II MHC transactivator (CIITA). The optimal combination was screened in vitro by flow cytometric analysis and mixed lymphocyte reaction assays and was validated in vivo in mouse models of leukemia and lymphoma. Functionality was assessed in an autologous setting using patient samples and in an allogeneic setting using an allogeneic mouse model. RESULTS: The combination of the Epstein-Barr virus TAPi and an shRNA targeting CIITA was efficient and effective at reducing cell surface MHC classes I and II in αCD19 'stealth' CAR-T cells while retaining in vitro and in vivo antitumor functionality. Mixed lymphocyte reaction assays and IFNγ ELISpot assays performed with T cells from patients previously treated with autologous αCD19 CAR-T cells confirm that CAR T cells expressing the stealth transgenes evade allogeneic and autologous anti-CAR responses, which was further validated in vivo. Importantly, we noted anti-CAR-T cell responses in patients who had received multiple CAR-T cell infusions, and this response was reduced on in vitro restimulation with autologous CARs containing the stealth transgenes. CONCLUSIONS: Together, these data suggest that the proposed stealth transgenes may reduce the immunogenicity of autologous and allogeneic cellular therapeutics. Moreover, patient data indicate that repeated doses of autologous FMC63-based αCD19 CAR-T cells significantly increased the anti-CAR T cell responses in these patients.

Changes in Brain Activity Immediately Post-Exercise Indicate a Role for Central Fatigue in the Volitional Termination of Exercise.

Electroencephalography (EEG) allows for the evaluation of real time changes in brain (electrocortical) activity during exercise. A few studies have examined changes in electrocortical activity using stationary cycling, but the findings have been mixed. Some of these studies have found increases in brain activity following exercise, while others have found decreases in brain activity following exercise. Hence, it is of importance to identify post-exercise changes in brain activity. Sixteen healthy, untrained subjects (8 males; 8 females) participated in the study. All 16 participants performed a graded exercise test (GXT) to volitional exhaustion on an upright cycle ergometer. Continuous EEG recordings were sampled before (PRE) and immediately following (IP) the GXT. Regions of interest were primarily the dorsolateral prefrontal cortex (DLPFC), ventrolateral prefrontal cortex (VLPFC), and left and right motor cortex (MC). In the DLPFC, a frontal asymmetry index was also identified. There was a statistically significant increase in theta power in the DLPFC, VLPFC, and left and right MC from PRE to IP (all p < 0.05). There was also a shift towards right hemisphere asymmetry at the IP time point in the DLPFC (p < 0.05). Finally, there was an increase in alpha power from PRE to IP in the right MC (p < 0.05). EEG could prove to be an important way to measure the effects of central fatigue on brain activity before and immediately following exercise.

Bioelectrochemically-assisted ammonia recovery from dairy manure.

The sustainability of direct land application of dairy manure is challenged by significant nutrient losses. Bioelectrochemical systems for ammonia recovery offer a manure management strategy that can recover both ammoniacal and organic nitrogen as a stable ammonia fertilizer. In this research, a microbial fuel cell (MFC) was used to treat two types of dairy manure under a variety of imposed anode compartment conditions. The system achieved a maximum coulombic efficiency of 20 ± 18 % and exhibited both COD and total nitrogen removals of approximately 60 %. Furthermore, the MFC showed a maximum organic nitrogen removal of 73.8 ± 12.1 %, and no differences in organic nitrogen (orgN) removal were detected among different conditions tested. Decreasing concentrations of anolyte ammonia nitrogen coupled with the observed orgN removal from the anolyte indicate that the MFC is effective at recovering orgN in dairy manure as ammoniacal nitrogen in the catholyte. Additionally, ion competition between NH4+ and other relevant cations (Na+, K+, and Mg2+) for transport across the CEM was investigated, with only K+ showing minor competitive effects. Based on the results of this research, we propose three key processes and two sub-processes that contribute to the successful operation of the MFC for nitrogen recovery from dairy manure. Bioelectrochemical systems for nitrogen recovery from dairy manure offer a novel, robust technology for producing a valuable ammonia nitrogen fertilizer, a thus far untapped resource in dairy manure streams.

Mesothelin CAR T Cells Secreting Anti-FAP/Anti-CD3 Molecules Efficiently Target Pancreatic Adenocarcinoma and its Stroma.

PURPOSE: Targeting solid tumors with chimeric antigen receptor (CAR) T cells remains challenging due to heterogenous target antigen expression, antigen escape, and the immunosuppressive tumor microenvironment (TME). Pancreatic cancer is characterized by a thick stroma generated by cancer-associated fibroblasts (CAF), which may contribute to the limited efficacy of mesothelin-directed CAR T cells in early-phase clinical trials. To provide a more favorable TME for CAR T cells to target pancreatic ductal adenocarcinoma (PDAC), we generated T cells with an antimesothelin CAR and a secreted T-cell-engaging molecule (TEAM) that targets CAF through fibroblast activation protein (FAP) and engages T cells through CD3 (termed mesoFAP CAR-TEAM cells). EXPERIMENTAL DESIGN: Using a suite of in vitro, in vivo, and ex vivo patient-derived models containing cancer cells and CAF, we examined the ability of mesoFAP CAR-TEAM cells to target PDAC cells and CAF within the TME. We developed and used patient-derived ex vivo models, including patient-derived organoids with patient-matched CAF and patient-derived organotypic tumor spheroids. RESULTS: We demonstrated specific and significant binding of the TEAM to its respective antigens (CD3 and FAP) when released from mesothelin-targeting CAR T cells, leading to T-cell activation and cytotoxicity of the target cell. MesoFAP CAR-TEAM cells were superior in eliminating PDAC and CAF compared with T cells engineered to target either antigen alone in our ex vivo patient-derived models and in mouse models of PDAC with primary or metastatic liver tumors. CONCLUSIONS: CAR-TEAM cells enable modification of tumor stroma, leading to increased elimination of PDAC tumors. This approach represents a promising treatment option for pancreatic cancer.

Delayed-Onset Muscle Soreness Alters Mechanical Sensitivity, but Not Thermal Sensitivity or Pain Modulatory Function.

Introduction: Many clinical musculoskeletal pain conditions are characterized by chronic inflammation that sensitizes nociceptors. An unresolved issue is whether inflammation affects all nociceptors in a similar manner. Exercise-induced muscle damage (EIMD) has been proposed as a model for simulating clinical inflammatory pain in healthy samples. We sought to test the effect of EIMD on various painful stimuli (pressure and thermal), central pain processing (via the nociceptive flexion reflex) and endogenous pain modulation via conditioned pain modulation and exercise-induced hypoalgesia. Methods: Eighteen participants (9F, age: 24.6 ± 3.3) were recruited for repeated measures testing and each completed pain sensitivity testing prior to and 48 hours after an eccentric exercise protocol. The participants performed a minimum of 6 rounds of 10 eccentric knee extension exercises to induce muscle damage and localized inflammation in the right quadriceps. Force decrements, knee range-of-motion, and delayed onset muscle soreness (DOMS) were used to quantify EIMD. Results: There was a significant main effect of time for pressure pain (%diff; -58.9 ± 23.1; p = 0.02, ηp2 = 0.28) but no significant main effect was observed for limb (%diff; -15.5 ± 23.9; p = 0.53, ηp2 = 0.02). In contrast, there was a significant interaction between time and limb (p < 0.001, ηp2 = 0.47) whereby participants had lower pressure pain sensitivity in the right leg only after the damage protocol (%diff; -105.9 ± 29.2; p = 0.002). Discussion: Individuals with chronic inflammatory pain usually have an increased sensitivity to pressure, thermal, and electrical stimuli, however, our sample, following muscle damage to induce acute inflammation only had sensitivity to mechanical pain. Exercise induced inflammation may reflect a peripheral sensitivity localized to the damaged muscle rather than a global sensitivity like those with chronic pain display.

Modeling ammonia emissions from manure in conventional, organic, and grazing dairy systems and practices to mitigate emissions.

Almost 60% of all ammonia (NH3) emissions are from livestock manure. Understanding the sources and magnitude of NH3 emissions from manure systems is critical to implement mitigation strategies. This study models 13 archetypical conventional (5 farms), organic (5 farms), and grazing (3 farms) dairy farms to estimate NH3 emissions from manure at the barn, storage, and after land application. Mitigation practices related to management of the herd, crop production, and manure are subsequently modeled to quantify the change in NH3 emissions from manure by comparing archetypical practices with these alternative practices. A mass balance of nutrients is also conducted. Emissions per tonne of excreted manure for the manure system (barn, storage, and land application) range from 3.0 to 4.4 g of NH3 for conventional farms, 3.5 to 4.4 g of NH3 for organic farms, and 3.4 to 3.9 g of NH3 for grazing farms. For all farm types, storage and land application are the main sources of NH3 emissions from manure. In general, solid manures have higher emission intensities due to higher pH during storage (pH = 7.4 for liquid, 7.8 for slurry, and 8.5 for solid manure) and lower infiltration rates after land application when compared with slurry and liquid manures. The most effective management practices to reduce NH3 emissions from manure systems are combining solid-liquid separation with manure injection (up to 49% reduction in NH3 emissions), followed by injection alone, and reducing crude protein in the dairy ration, especially in organic and grazing farms that have grazing and forages as the main component of the dairy ration. This study also shows that the intensity of NH3 emissions from manure depends significantly on the functional unit and presents results per manure excreted, total solids in excreted manure, animal units, and fat- and protein-corrected milk.

Soil greenhouse gas flux and nitrogen mineralization following manure application from tannin-fed dairy cows.

Growing concerns about environmental impacts of dairy farms have driven producers to address greenhouse gas (GHG) emissions and nitrogen (N) losses from soil following land application of dairy manure. Tannin dietary additives have proved to be a successful intervention for mitigating GHG and ammonia (NH3 ) emissions at the barn scale. However, it is unknown how land application of dairy manure from cows fed tannin diets affects crop-soil nitrogen dynamics and soil GHG flux. To test this, cows were fed diets at three levels of tannins (0.0%, 0.4%, and 1.8% of dry matter intake) and their manure was field applied at two N rates (240 and 360 kg N ha-1 ). Soil NH4 + -N, NO3 - -N, corn silage yield, and soil GHG flux were then measured over a full growing season. Soils amended with tannin manure had lower initial NH4 + -N concentrations and lower total mineral N (NH4 + -N + NO3 - -N) concentrations 19 days after application, compared to soils amended with no tannin manures. Despite lower early season N availability in tannin-fertilized plots, there were no differences in corn silage yield. No differences in soil GHG and NH3 emissions were observed between manure-amended treatments. These results demonstrate that while tannin addition to dairy cow feed does not offer short-term GHG or NH3 emissions reductions after field manure application, it can promote slower soil N mineralization that may reduce reactive N loss after initial application.

Ankle dorsiflexion asymmetry and the relationship with walking performance in people with multiple sclerosis.

BACKGROUND: Multiple sclerosis (MS) is a neurological disease characterized by demyelination disrupting the central nervous system. Persons with MS may exhibit symptomatic strength asymmetry (SA) that impacts motor gait and ankle mobility. The purpose of the present study was to investigate ankle dorsiflexion SA in people with MS and its relationship to functional performance. RESEARCH QUESTION: Is their a difference in dorsiflexion SA in MS participants compared to healthy individuals and does it impact functional performance? METHODS: 13 MS participants (EDSS 3.5 + 1.8) and 13 age matched NON-MS participants underwent maximal isometric (MVC) dynamometry testing for ankle dorsiflexion in both limbs to determine SA. Participants performed three functional tasks of walking performance. RESULTS: There was a significant intra-limb MVC difference in the MS group, and significantly greater isometric SA (p < 0.007) and isokinetic SA (p < 0.04) in the MS group compared to healthy individuals. The MS group exhibited significant correlations between outcomes of functional walking performance with isokinetic but not isometric SA. There was no significant correlation between disability status and functional task performance. SIGNIFICANCE: Ankle dorsiflexion SA is negatively correlated with functional performance in MS participants. MS disability status was not a predictor of functional task performance, and symptom testing may be appropriate to assess walking ability in persons with MS.

Chemical genetic control of cytokine signaling in CAR-T cells using lenalidomide-controlled membrane-bound degradable IL-7.

CAR-T cell therapy has emerged as a breakthrough therapy for the treatment of relapsed and refractory hematologic malignancies. However, insufficient CAR-T cell expansion and persistence is a leading cause of treatment failure. Exogenous or transgenic cytokines have great potential to enhance CAR-T cell potency but pose the risk of exacerbating toxicities. Here we present a chemical-genetic system for spatiotemporal control of cytokine function gated by the off-patent anti-cancer molecular glue degrader drug lenalidomide and its analogs. When co-delivered with a CAR, a membrane-bound, lenalidomide-degradable IL-7 fusion protein enforced a clinically favorable T cell phenotype, enhanced antigen-dependent proliferative capacity, and enhanced in vivo tumor control. Furthermore, cyclical pharmacologic combined control of CAR and cytokine abundance enabled the deployment of highly active, IL-7-augmented CAR-T cells in a dual model of antitumor potency and T cell hyperproliferation.

Leukemia-intrinsic determinants of CAR-T response revealed by iterative in vivo genome-wide CRISPR screening.

CAR-T therapy is a promising, novel treatment modality for B-cell malignancies and yet many patients relapse through a variety of means, including loss of CAR-T cells and antigen escape. To investigate leukemia-intrinsic CAR-T resistance mechanisms, we performed genome-wide CRISPR-Cas9 loss-of-function screens in an immunocompetent murine model of B-cell acute lymphoblastic leukemia (B-ALL) utilizing a modular guide RNA library. We identified IFNγR/JAK/STAT signaling and components of antigen processing and presentation pathway as key mediators of resistance to CAR-T therapy in vivo; intriguingly, loss of this pathway yielded the opposite effect in vitro (sensitized leukemia to CAR-T cells). Transcriptional characterization of this model demonstrated upregulation of these pathways in tumors relapsed after CAR-T treatment, and functional studies showed a surprising role for natural killer (NK) cells in engaging this resistance program. Finally, examination of data from B-ALL patients treated with CAR-T revealed an association between poor outcomes and increased expression of JAK/STAT and MHC-I in leukemia cells. Overall, our data identify an unexpected mechanism of resistance to CAR-T therapy in which tumor cell interaction with the in vivo tumor microenvironment, including NK cells, induces expression of an adaptive, therapy-induced, T-cell resistance program in tumor cells.

Anti-TACI single and dual-targeting CAR T cells overcome BCMA antigen loss in multiple myeloma.

Chimeric Antigen Receptor (CAR) T cells directed to B cell maturation antigen (BCMA) mediate profound responses in patients with multiple myeloma, but most patients do not achieve long-term complete remissions. In addition, recent evidence suggests that high-affinity binding to BCMA can result in on-target, off-tumor activity in the basal ganglia and can lead to fatal Parkinsonian-like disease. Here we develop CAR T cells against multiple myeloma using a binder to targeting transmembrane activator and CAML interactor (TACI) in mono and dual-specific formats with anti-BCMA. These CARs have robust, antigen-specific activity in vitro and in vivo. We also show that TACI RNA expression is limited in the basal ganglia, which may circumvent some of the toxicities recently reported with BCMA CARs. Thus, single-targeting TACI CARs may have a safer toxicity profile, whereas dual-specific BCMA-TACI CAR T cells have potential to avoid the antigen escape that can occur with single-antigen targeting.

Confirmation and refutation of very luminous galaxies in the early Universe.

During the first 500 million years of cosmic history, the first stars and galaxies formed, seeding the Universe with heavy elements and eventually reionizing the intergalactic medium1-3. Observations with the James Webb Space Telescope (JWST) have uncovered a surprisingly high abundance of candidates for early star-forming galaxies, with distances (redshifts, z), estimated from multiband photometry, as large as z ≈ 16, far beyond pre-JWST limits4-9. Although such photometric redshifts are generally robust, they can suffer from degeneracies and occasionally catastrophic errors. Spectroscopic measurements are required to validate these sources and to reliably quantify physical properties that can constrain galaxy formation models and cosmology10. Here we present JWST spectroscopy that confirms redshifts for two very luminous galaxies with z > 11, and also demonstrates that another candidate with suggested z ≈ 16 instead has z = 4.9, with an unusual combination of nebular line emission and dust reddening that mimics the colours expected for much more distant objects. These results reinforce evidence for the early, rapid formation of remarkably luminous galaxies while also highlighting the necessity of spectroscopic verification. The large abundance of bright, early galaxies may indicate shortcomings in current galaxy formation models or deviations from physical properties (such as the stellar initial mass function) that are generally believed to hold at later times.

Cerebral blood flow dynamics: Is there more to the story at exercise onset?

A monoexponential model characterizing cerebral blood velocity dynamics at the onset of exercise may mask dynamic responses by the cerebrovasculature countering large fluctuations of middle cerebral artery blood velocity (MCAv) and cerebral perfusion pressure (CPP) oscillations. Therefore, the purpose of this study was to determine whether the use of a monoexponential model attributes initial fluctuations of MCAv at the start of exercise as a time delay (TD). Twenty-three adults (10 women, 23.9 ± 3.3 yrs; 23.7 ± 2.4 kg/m2 ) completed 2 min of rest followed by 3 mins of recumbent cycling at 50 W. MCAv, CPP, and Cerebrovascular Conductance index (CVCi), calculated as CVCi = MCAv/MAP × 100 mmHg, were collected, a lowpass filter (0.2 Hz) was applied, and averaged into 3-second bins. MCAv data were then fit to a monoexponential model [ΔMCAv(t) = Amp(1 - e-(t-TD)/τ )]. TD, tau (τ), and mean response time (MRT = TD + τ) were obtained from the model. Subjects exhibited a TD of 20.2 ± 18.1 s. TD was directly correlated with MCAv nadir (MCAvN ), r = -0.560, p = 0.007, which occurred at similar times (16.5 ± 15.3 vs. 20.2 ± 18.1 s, p = 0.967). Regressions indicated CPP as the strongest predictor of MCAvN ( R a 2 $$ {R}_a^2 $$ = 0.36). Fluctuations in MCAv were masked using a monoexponential model. To adequately understand cerebrovascular mechanisms during the transition from rest to exercise, CPP and CVCi must also be analyzed. A concurrent drop in cerebral perfusion pressure and middle cerebral artery blood velocity at the start of exercise forces the cerebrovasculature to respond to maintain cerebral blood flow. The use of a monoexponential model characterizes this initial phase as a time delay and masks this large important response.

Tandem chimeric antigen receptor (CAR) T cells targeting EGFRvIII and IL-13Rα2 are effective against heterogeneous glioblastoma.

Background: Chimeric antigen receptor (CAR) T cells have achieved remarkable responses in patients with hematological malignancies; however, the potential of this therapeutic platform for solid tumors like glioblastoma (GBM) has been limited, due in large part to the targeting of single antigens in a heterogeneous disease. Strategies that allow CAR T cells to engage multiple antigens concomitantly may broaden therapeutic responses and mitigate the effects of immune escape. Methods: Here we have developed a novel, dual-specific, tandem CAR T (TanCART) cell with the ability to simultaneously target both EGFRvIII and IL-13Rα2, two well-characterized tumor antigens that are frequently found on the surface of GBM cells but completely absent from normal brain tissues. We employed both standard immunological assays and multiple orthotopic preclinical models including patient-derived xenograft to demonstrate efficacy of this approach against heterogeneous tumors. Results: Tandem CAR T cells displayed enhanced cytotoxicity in vitro against heterogeneous GBM populations, including patient-derived brain tumor cultures (P < .05). Compared to CAR T cells targeting single antigens, dual antigen engagement through the tandem construct was necessary to achieve long-term, complete, and durable responses in orthotopic murine models of heterogeneous GBM, including patient-derived xenografts (P < .05). Conclusions: We demonstrate that TanCART is effective against heterogeneous tumors in the brain. These data lend further credence to the development of multi-specific CAR T cells in the treatment of GBM and other cancers.

Systematic Interrogation of Tumor Cell Resistance to Chimeric Antigen Receptor T-cell Therapy in Pancreatic Cancer.

Chimeric antigen receptor (CAR) T-cell therapy can lead to dramatic clinical responses in B-cell malignancies. However, early clinical trials with CAR T-cell therapy in non-B-cell malignancies have been disappointing to date, suggesting that tumor-intrinsic features contribute to resistance. To investigate tumor-intrinsic modes of resistance, we performed genome scale CRISPR-Cas9 screens in mesothelin (MSLN)-expressing pancreatic cancer cells. Co-culture with MSLN-targeting CAR T cells identified both antigen-dependent and antigen-independent modes of resistance. In particular, loss of the majority of the genes involved in the pathway responsible for GPI-anchor biosynthesis and attachment abrogated the ability of CAR T cells to target pancreatic cancer cells, suggesting that disruption of this pathway may permit MSLN CAR T-cell evasion in the clinic. Antigen-independent mediators of CAR T-cell response included members of the death receptor pathway as well as genes that regulate tumor transcriptional responses, including TFAP4 and INTS12. TFAP4-mediated CAR T resistance depended on the NFκB transcription factor p65, indicating that tumor resistance to CAR T-cell therapy likely involves alterations in tumor-intrinsic states. Overall, this study uncovers multiple antigen-dependent and -independent mechanisms of CAR T-cell evasion by pancreatic cancer, paving the way for overcoming resistance in this disease that is notoriously refractory to immunotherapy. SIGNIFICANCE: The identification and validation of key determinants of CAR T-cell response in pancreatic cancer provide insights into the landscape of tumor cell intrinsic resistance mechanisms and into approaches to improve therapeutic efficacy.

Four challenges to CAR T cells breaking the glass ceiling.

Cell-based therapies using chimeric antigen receptor T cells (CAR T) have had dramatic efficacy in the clinic and can even mediate curative responses in patients with hematologic malignancies. As living drugs, engineered cells can still be detected in some patients even years after the original infusion. The excitement around the cell therapy field continues to expand as recent reports have shown that CAR T cells can induce remission in patients with autoimmune disease. While these promising advances in the field garner hope for wide-spread utility of CAR T therapies across diseases, several roadblocks exist that currently limit the access and efficacy of this therapy in the clinic. Herein, we will discuss four major obstacles that the CAR T field faces, including toxicity, identifying tumor-specific antigens, improving function in solid tumors, and reducing manufacturing complexity and cost. CAR T cells have potential for a multitude of diseases, but these glass ceilings will need to be broken in order to improve clinical responses and make this potentially life-saving therapy accessible to a larger patient population.

Higher rating of perceived exertion and lower perceived recovery following a graded exercise test during menses compared to non-bleeding days in untrained females.

The underrepresentation of the female population in exercise sciences could be attributed, at least in part, to difficulty in appropriately accounting for the effects of the menstrual cycle (MC). Previous studies examining the effects of the MC on aerobic performance and subjective measures of aerobic performance show conflicting results. Purpose: The study examined how the MC affects the objective and subjective measures of aerobic performance within untrained female participants and in comparison with untrained male participants assessed at similar time intervals. Methods: Twenty-one participants (12 females and 9 males) completed a graded exercise test (GXT) on a cycle ergometer. The female participants were tested during their early follicular (EF; menses), ovulatory (O), and mid-luteal (ML) phases of the MC. The male participants were included as the control group and were randomly assigned to a menstrual cycle phase for each visit. During GXT, maximal oxygen consumption (VO2max), respiratory exchange ratio (RER), maximal heart rate (HRmax), peak blood lactate, and rating of perceived exertion (RPE) were determined. Twenty-four hours post-exercise, the perceived recovery status (PRS) was assessed. The MC phase was estimated using basal body temperature (BBT) in the female participants. Results: The male participants obtained a higher peak power and VO2max compared to the female participants (p < 0.05). All objective measures of aerobic performance did not significantly differ across the MC phases or time points that were tested. In the untrained female participants, an effect of the MC phase on RPE was found, with RPE being higher at EF (8.92 ± 0.79) compared to O (7.67 ± 1.23; p < 0.05) and ML (7.75 ± 1.06; p < 0.05). In addition, an effect of the MC phase on PRS was found, with perceived recovery being lower at EF (6.83 ± 0.94) compared to O (8.83 ± 1.12) and ML (8.67 ± 0.65; all p < 0.005) for the untrained female participants. No significant differences in RPE and PRS were found between tests in the untrained male participants. The female participants had lower perceived recovery following EF (6.83 ± 0.94) compared with the male participants (9.00 ± 1.00; p < 0.001). Conclusion: The untrained female participants perceived greater exertion during GXT and impaired recovery following GXT in EF compared to O and ML. These results may be attributed to either a drop in female sex hormone concentrations or discomfort associated with menses. The male participants did not exhibit any changes over time. Future studies using subjective parameters such as perceived exertion to track the internal load of training in the naturally menstruating female population should consider menses.

Lean Mass is Associated with, but Does Not Mediate Sex Differences in Pressure Pain Sensitivity in Healthy Adults.

Background: Sex differences exist in pain sensitivity, however, the underlying mechanism(s) that explain these differences are not fully understood. Pain sensitivity has been shown to be influenced by body mass index, but limited data exist on the role of body composition on pain sensitivity. The purpose was to examine the influence of body composition on pain sensitivity in males and females. Methods: This cross-sectional study design used pressure pain thresholds (PPT) of 87 participants (45 female) who were assessed in the vastus lateralis (leg PPT) and brachioradialis (arm PPT) using a pressure algometer. Fat and lean tissue were assessed via dual-energy X-ray absorptiometry (DXA). A two group by two limb, repeated measured ANOVA was used to assess differences between limbs and sex. Spearman correlations and hierarchical regression analyses were employed to determine the association between body composition and PPT. Results: Males had higher PPTs then females (P<0.05) and had higher DXA assessed lean and lower levels fat mass (P<0.05). Total body and limb specific lean mass was associated with PPTs (r≥0.34; P<0.05). Hierarchical regression analysis revealed lean mass was a significant predictor of 8% of the variance in arm PPT (P<0.006) and 18% of the variance in leg PPT (P<0.001). However, lean mass was not found to statistically mediate the observed sex differences in PPT. Conclusion: This finding suggests lean mass may play a previously unknown role in sex differences in pressure pain sensitivity. Future studies are needed to confirm this finding and a larger sample size is likely required to have sufficient power to perform the mediation analysis.

Noninflammatory presentation of cutaneous breast cancer: a retrospective case series at a single academic institution with review of the literature.

Breast cancer with skin involvement is an uncommon clinical presentation of this malignancy. Breast cancer overall has a relatively high mortality rate and wide variety of presentations, making skin involvement by breast cancer a necessary consideration in the differential diagnosis for many types of breast lesions. A retrospective review of our own small academic dermatology outpatient clinic, between August 2006 and January 2020, found four cases of noninflammatory breast cancer with skin involvement diagnosed through biopsy by our dermatologists. This review was approved by the local Institutional Review Board. Of the four patients identified, three were female and one was male. One patient had prior history of invasive ductal carcinoma in remission before recurrence was diagnosed. Another patient had a history of melanoma in situ before diagnosis with breast cancer. Patients were treated with various combinations of surgery, radiation, and hormone therapy. These four cases are presented here in detail, which emphasize the role of the dermatologist in recognizing various cutaneous manifestations of noninflammatory breast cancer in order to make a timely diagnosis.