Transcriptomics, proteomics, and physiological assays reveal immunosuppression in the eastern oyster Crassostrea virginica exposed to acidification stress.

Ocean acidification (OA) is recognized as a major stressor for a broad range of marine organisms, particularly shell-building invertebrates. OA can cause alterations in various physiological processes such as growth and metabolism, although its effect on host-pathogen interactions remains largely unexplored. In this study, we used transcriptomics, proteomics, and physiological assays to evaluate changes in immunity of the eastern oyster Crassostrea virginica exposed to OA conditions (pH = 7.5 vs pH = 7.9) at various life stages. The susceptibility of oyster larvae to Vibrio infection increased significantly (131 % increase in mortality) under OA conditions, and was associated with significant changes in their transcriptomes. The significantly higher mortality of larvae exposed to pathogens and acidification stress could be the outcome of an increased metabolic demand to cope with acidification stress (as seen by upregulation of metabolic genes) at the cost of immune function (downregulation of immune genes). While larvae were particularly vulnerable, juveniles appeared more robust to the stressors and there were no differences in mortality after pathogen (Aliiroseovarius crassostrea and Vibrio spp.) exposure. Proteomic investigations in adult oysters revealed that acidification stress resulted in a significant downregulation of mucosal immune proteins including those involved in pathogen recognition and microbe neutralization, suggesting weakened mucosal immunity. Hemocyte function in adults was also impaired by high pCO2, with a marked reduction in phagocytosis (67 % decrease in phagocytosis) in OA conditions. Together, results suggest that OA impairs immune function in the eastern oyster making them more susceptible to pathogen-induced mortality outbreaks. Understanding the effect of multiple stressors such as OA and disease is important for accurate predictions of how oysters will respond to future climate regimes.

The Confusion Assessment Method in action: Implementation of a protocol to increase delirium screening and diagnosis.

The use of the Confusion Assessment Method (CAM) for delirium assessment in real-life can be inconsistent. We examined the impact of a protocol on delirium screening and detection in hospitalized older adults using the CAM. We analyzed data from 32,338 admissions to a quaternary hospital between 2018 and 2022. We assessed the percentage of admissions screened for delirium, adherence to daily screening, positive screening, and overlap with ICD-10 coding. The percentage of admissions screened for delirium increased from 74% in 2018 to 98.7% in 2022. Adherence to daily screening was achieved in 24.5% of admissions, and the percentage of positive screenings fluctuated between 8.4% and 11.5%. Among the admissions with a delirium-related ICD-10 code, 32% had a positive screening, 62% were negative, and 6% remained unscreened. While implementing a protocol increased the proportion of admissions screened for delirium, adherence to daily screening and consistency of positive delirium screenings remain areas for improvement.

Does an Adjusted Kinematic Model Predict the Relative Eccentric Force During Nordic Curl?

This study aimed to assess the combination of video-based kinematic variables adjusted by intrinsic covariates to predict the relative eccentric force (RelF) during the Nordic curl. The participants (n = 21) performed Nordic curls (3 trials; 3-min rest) on a device measuring the eccentric force. The peaks were normalized by body weight. Kinovea software was used to track angular and linear velocity and acceleration from recorded videos. Two prediction models with multiple linear regression equations associated kinematic, anthropometric, and age variables to adjust the actual RelF. The equations obtained the predicted RelF. The actual RelF was inversely correlated with height (r = -.52), tangential (r = -.50) and centripetal accelerations (r = -.715), and angular velocity (r = -.70). The best prediction models combined angular velocity with age (F2,18 = 15.1, P = .001, r = .792, r2 = .627) and with height (F2,18 = 14.5, P = .001, r = .785, r2 = .616). No differences were observed between actual and predicted values (P = .993-.994), with good levels of agreement and consistency (intraclass correlation coefficient = .77-.78; Cronbach α = .86-.87). Bland-Altman results showed high levels of agreement and low biases. The standard error of measurement and minimal detectable change ranges were 0.46 to 0.49 N/kg and 1.28 to 1.36 N/kg, respectively. Also, the percentage of standard error of measurement was below 10% (7.92%-8.35%). The coefficient of variation analysis returned a 14.54% and 15.13% for each model, respectively. Kinematic analysis offers portability and low cost to current expensive or technical impaired dynamometry-based techniques to assess the RelF.

Test-Retest of a Novel Wii Board-Based Device to Assess Eccentric Knee Flexor Strength During the Nordic Hamstring Exercise.

This study aimed to assess the test-retest reliability of a Wii board-based device to assess the eccentric knee strength. Healthy participants (n = 20, 22.7 [3.4] y, 1.71 [0.09] m, 69 [13] kg) performed 2 assessments-days 1 and 2-of the Nordic hamstring exercise (3 trials per day, 3 min of rest between trials, and 48 h between trials) on a padded board with an attached Wii Balance Board (WBB). A ratchet inelastic strap made the contact between the participant's distal aspect of both legs and the inverted upper surface of the WBB. The means among 3 trials were used to extract the absolute strength and the left and the right limb strengths. No between-session differences were found (P range = .691-.981). The intraclass correlation coefficient range showed excellent results (.905-.926), as the Cronbach α test (above .94). The correlation was high (r > .91; .820 < r2 < .862). The Bland-Altman analysis returned high levels of agreement. The standard error of measurement ranged from 5.56 to 11.07 N and the minimal detectable change from 15.42 to 30.68 N. The percentage of standard error of measurement values were 2.95%, 3.74%, 2.88%, respectively, for absolute strength, right limb strength, and left limb strength. The adapted system showed an excellent cost-benefit relationship with optimal test-retest reliability. The findings suggest that the adapted system, using the WBB, is a reliable method for measuring the eccentric knee strength.

Overhead Shoulder Position Versus Scapular Retraction Position to Normalize the Electromyographic Signal of the Lower Trapezius Muscle.

This study aimed to compare electromyographic response of the lower trapezius muscle during maximal voluntary isometric contractions under two conditions: overhead shoulder position versus scapular position, rating the pain after each condition. Twenty-six participants with shoulder pain (SP; n = 15; Shoulder Pain and Disability Index-Pain = 34 [range 6-66]) and without shoulder pain (nSP; n = 11; Shoulder Pain and Disability Index-Pain = 0 [range 0-14]) performed maximal voluntary isometric contractions in those 2 conditions while the lower trapezius was monitored using surface electromyography. Pain was rated using the visual analog scale. The nSP group experienced no pain, whereas the SP group reported pain in both conditions with higher pain during the overhead movement. No differences were observed for muscle excitation between conditions, but higher levels were observed for the nSP group compared with the SP group. The overhead condition was more painful for the SP group (visual analog scale: SP = 4 vs nSP = 1.09, P = .036). Scapular position showed the same levels of LT excitation than overhead position. The scapular position is preferable, compared with overhead positioning, to normalize the lower trapezius instead of overhead positioning.

Acute Effects of Single- Versus Double-Leg Postactivation Potentiation on Postural Balance of Older Women: An Age-Matched Controlled Study.

AIMS: To compare the postactivation potentiation effects of isometric contraction until failure in double- and single-leg tasks on older women's balance. METHODS: The one-legged balance test was performed before and immediately after a rise-to-toes task until the task failure. Older women were divided into two groups: a group performed the task with double leg (n = 43) and the other group with single-leg support (n = 55). RESULTS: The single-leg group showed slower velocity of sway post rise-to-toes task (pre = 4.02 ± 1; post = 3.78 ± 1.15 m/s; p = .04) without differences for the center of pressure path length (pre = 79 ± 21; post = 75 ± 23 cm; p = .08). In the double-leg group, faster velocity of sway (pre = 4 ± 1.22; post = 4.25 ± 1.13; p = .03) and increased center of pressure path length (pre = 80 ± 24; post = 85 ± 23 cm; p = .03) were observed after the task. CONCLUSIONS: The single-leg group showed improved balance outcomes due to postactivation potentiation, while the double-leg group showed worsened balance consistent with muscle fatigue.

Parasite Infections, Allergy and Asthma: A Role for Tropomyosin in Promoting Type 2 Immune Responses.

INTRODUCTION: The relationship of parasite infections and promotion or protection from allergy and asthma is controversial. Currently, over 1.5 billion people are infected with parasites worldwide, and Ascaris lumbricoides is the most frequent soil-transmitted helminth. OBJECTIVES: To evaluate the biological activity of recombinant A. lumbricoides tropomyosin and investigate IgE cross-reactive responses to tropomyosins by means of microarray methodology for the detection of sensitization to allergen components. METHODS: Forty patients 12-75 years of age (25 males) with asthma and/or rhinitis and 10 nonallergic control subjects participated in this study. All patients presented positive skin tests to cockroach extracts and underwent skin prick testing (SPT) with recombinant (r) tropomyosins rPer a 7 from Periplaneta americana and rAsc l 3 from A. lumbricoides, at 10 µg/mL. IgE to cockroach and parasite tropomyosins were measured by chimeric ELISA and ImmunoCAP-ISAC, and total IgE was quantitated by ImmunoCAP. Agreement of results was assessed by κ statistics. RESULTS: Recombinant A. lumbricoides showed biological activity, inducing positive skin tests in 50% patients with asthma and/or rhinitis. IgE to cockroach and parasite tropomyosins were detected in 55-62% of patients. There was good-to-excellent agreement of results of SPT and IgE measurements by ELISA and ImmunoCAP-ISAC, with κ indices of 0.66-0.95. No skin test reactivity or IgE antibodies to tropomyosins were found in nonallergic individuals. CONCLUSIONS: Our results suggest that IgE responses to tropomyosin from A. lumbricoides may enhance reactivity to homologous allergens upon exposure by inhalation or ingestion, promoting allergic reactions and asthma, or increasing the severity of these clinical conditions.

Effects of 4-Week Diacutaneous Fibrolysis on Myalgia, Mouth Opening, and Level of Functional Severity in Women With Temporomandibular Disorders: A Randomized Controlled Trial.

OBJECTIVE: The purpose of this study was to assess the effects of 4-week protocol of diacutaneous fibrolysis (DF) compared with simulated DF (sham-DF) on myalgia and mouth opening. METHODS: In a sham randomized controlled trial, 34 women with temporomandibular disorders and myofascial pain were randomly divided as intervention group (IG) and sham-DF group (SG). The IG received 4 weeks of real DF, and the SG received sham. Pain was assessed through the visual analog scale and pressure pain thresholds (PPTs) on the temporomandibular joint (TMJ), and over the temporal and masseter muscles. The Mandibular Function Impairment Questionnaire was used to classify the participants regarding to the severity of the functional limitation related to TMD. RESULTS: Pain scores decreased for both groups, but the IG showed lower values at week 4, with between-group differences. Bilateral temporal PPT showed higher values at week 4, with between-group differences. The SG had lower PPTs but the IG had higher PPTs, both compared to baseline results. The time-by-group interaction and the frequency of participants above 40 mm of mouth opening showed a significant difference for the IG over time with higher results at the 4-week assessment compared to its own baseline. Both groups showed lower MFIQ scores from baseline to 4-week assessment. There was a lower frequency of a moderate level of severity for the IG. No differences were observed for TMJ or for the masseter muscles PPT. CONCLUSION: Improvements were observed for visual analog scale scores and PPTs on temporal muscles. There was a group-by-time interaction in the IG, suggesting a possible potential use of DF for mouth opening.

Validity of an Inexpensive Hanging Scale During Isometric Shoulder Movements.

CONTEXT: Strength assessment is essential to prescribe exercise in sports and rehabilitation. Low-cost valid equipment may allow continuous monitoring of training. OBJECTIVE: To examine the validity of a very low-cost hanging scale by comparing differences in the measures of peak force to a laboratory grade load cell during shoulder abduction, flexion, extension, and internal and external rotations. DESIGN: Analytical study. PARTICIPANTS: Thirty-two healthy subjects (18 women, age 26 [10] y, height 172 [8] cm, mass 69 [13] kg, body mass index 23 [4] kg/m2). MAIN OUTCOME MEASURES: The dependent variable was the maximal peak force (in kilogram-force). The independent variable was the instrument (laboratory grade load cell and hanging scale). RESULTS: No differences were observed while comparing the results. The intraclass correlation coefficients1,1 ranged from .96 to .99, showing excellent results. The Cronbach alpha test also returned >.99 for all comparisons. The SEM ranged from 0.02 to 0.04 kgf, with an averaged SD from 0.24 to 0.38 kgf. The correlation was classified as high for all tested movements (r > .99; P < .001), with excellent adjusted coefficients of determination (.96 < r2 < .99). Bland-Altman results showed high levels of agreement with bias ranging from 0.27 to 0.48. CONCLUSIONS: Hanging scale provides valid measures of isometric strength with similar output measures as laboratory grade load cell.

Effects of 8 weeks of masticatory muscles focused endurance exercises on women with oro-facial pain and temporomandibular disorders: A placebo randomised controlled trial.

BACKGROUND: Exercises are used to treat temporomandibular disorders (TMD), but they are often assessed with other therapies. Local endurance exercises may alter the resistance to fatigue and pain. OBJECTIVE: To assess the effects of an 8-week protocol of local endurance exercises of masticatory muscles on muscle excitation, force response, perceived pain and over muscle efficiency. DESIGN: Randomised controlled trial. SETTING: Ambulatory care. SUBJECTS: In a placebo randomised controlled trial, 46 women with TMD and oro-facial pain were randomised into intervention group and placebo group. The intervention group received a protocol of biting endurance exercises, controlled by biofeedback. The placebo group received a placebo (simulated laser therapy). MAIN OUTCOME MEASURES: The primary outcomes were collected at baseline, 4 weeks and 8 weeks. Pain was assessed through visual analogue scale (VAS) and pressure pain thresholds (PPT). Bite force was collected by a load cell synchronised with surface electromyography of masticatory muscles, bilaterally. RESULTS: Pain scores decreased for both groups, but the intervention group showed lower values at 8 weeks. No differences were noted between groups for PPT, but the results increased for both overtime. Time until fatigue and muscle efficiency were higher in the intervention group vs placebo group in both within- and between-subject analysis. Force increased from 4 to 8 weeks in the PG, without differences between groups. Temporal muscle excitation was higher on 8 weeks compared with baseline for the intervention group, without differences between groups. CONCLUSION: Eight-week exercise protocol of muscle endurance alleviates the pain and improves the resistance to fatigue and muscle efficiency in TMD subjects.

Pain, balance, grip strength and gait parameters of older adults with and without post-chikungunya chronic arthralgia.

OBJECTIVE: To compare the levels of pain, grip strength, balance and gait in older adults with and without post-Chikungunya chronic arthralgia (PCCA). METHODS: Sixty-two older adults, 30 with and 32 without PCCA participated in the study. Pain level was assessed using a Visual Analogue Scale (VAS). Gait during a 10-m walk was assessed using inertial sensors. Semi-static balance was assessed during an eyes-closed bipedal balance test on a force platform, and grip strength was assessed using a hand dynamometer. RESULTS: Participants with PCCA presented severe levels of pain (VAS > 7.5), poorer balance, lower grip strength, walked slower, with lower cadence and stride length and higher stride time and stride length variability than participants without PCCA (P < 0.001 for all variables). CONCLUSIONS: Older adults with PCCA had high levels of pain, impaired balance and gait and lower grip strength compared to older adults without PCCA.

Regulation of apoptosis-related genes during interactions between oyster hemocytes and the alveolate parasite Perkinsus marinus.

The alveolate Perkinsus marinus is the most devastating parasite of the eastern oyster Crassostrea virginica. The parasite is readily phagocytosed by oyster hemocytes, but instead of intracellular killing and digestion, P. marinus can survive phagocytosis and divide in host cells. This intracellular parasitism is accompanied by a regulation of host cell apoptosis. This study was designed to gain a better understanding of the molecular mechanisms of apoptosis regulation in oyster hemocytes following exposure to P. marinus. Regulation of apoptosis-related genes in C. virginica, and apoptosis-regulatory genes in P. marinus, were investigated via qPCR to assess the possible pathways involved during these interactions. In vitro experiments were also carried out to evaluate the effect of chemical inhibitors of P. marinus antioxidant processes on hemocyte apoptosis. Results indicate the involvement of the mitochondrial pathway (Bcl-2, anamorsin) of apoptosis in C. virginica exposed to P. marinus. In parallel, the antioxidants peroxiredoxin and superoxide dismutase were regulated in P. marinus exposed to C. virginica hemocytes suggesting that apoptosis regulation in infected oysters may be mediated by anti-oxidative processes. Chemical inhibition of P. marinus superoxide dismutase resulted in a marked increase of reactive oxygen species production and apoptosis in infected hemocytes. The implication of oxygen-dependent apoptosis during P. marinus infection and disease development in C. virginica is discussed.

Recombinant allergens for diagnosis of cockroach allergy.

Molecular cloning of cockroach allergens and their expression as recombinant proteins have allowed a better understanding of the mechanisms of cockroach allergic disease. Recombinant cockroach allergens have been used for skin testing or in vitro methods to measure IgE antibody levels in serum. Early studies evaluating selected U.S. patients revealed that a cocktail of four cockroach allergens, Bla g 1, Bla g 2, Bla g 4, and Bla g 5, would identify 95 % of cockroach allergic patients. More recent studies pointed to an important role of sensitization to tropomyosin among certain populations, and suggested that a cocktail of five allergens Bla g 1 and/or Per a 1, Bla g 2, Bla g 4, Bla g 5, and Bla g 7, and/or Per a 7, would be expected to diagnose 50- 64 % of cockroach-allergic patients worldwide. Variation in IgE reactivity profiles could be in part due to IgE responses to cross-reactive homologous allergens from different origins. The availability of purified natural or recombinant cockroach allergens provides the capacity to improve diagnosis of cockroach allergy and to develop novel forms of immunotherapy for cockroach-allergic patients.

Efficacy of recombinant allergens for diagnosis of cockroach allergy in patients with asthma and/or rhinitis.

BACKGROUND: Immunoglobulin E (IgE) reactivity to individual allergens among cockroach-allergic patients has revealed wide variability. The aim of this study was to assess the effectiveness of recombinant cockroach allergens for skin testing, and to determine sensitization profiles among cockroach-allergic patients living in Brazil. METHODS: Fifty-seven cockroach-allergic patients with asthma and/or rhinitis were recruited. Skin testing with recombinant (r) allergens from Periplaneta americana (rPer a 1 and rPer a 7) and Blattella germanica (rBla g 2, rBla g 4 and rBla g 5) were performed at 10 µg/ml and 5 µg/ml (rPer a 1). IgE antibodies to rPer a 7 and rPer a 1 were quantitated by ELISA. RESULTS: Of 57 patients tested, 3 (5.3%), 24 (42.1%), 4 (7%), 3 (5.3%) and 4 (7%) showed positive reactions to rPer a 1, rPer a 7, rBla g 2, rBla g 4 and rBla g 5, respectively. Twenty-eight patients (49.1%) had positive tests to at least one allergen. In keeping with skin test results, 31/57 patients (54.4%) and 5/55 patients (9%) had detectable IgE to rPer a 7 and rPer a 1, respectively. Levels of IgE to rPer a 7 were higher in patients with positive tests to rPer a 7 than those with negative tests (geometric mean 13.2 and 1.8 IU/ml, p < 0.05). There was good concordance of results of skin tests and measurements of serum IgE to rPer a 7. CONCLUSION: IgE reactivity to rPer a 7 (P. americana tropomyosin) was dominant among patients in Brazil. However, 50% of the patients did not present reactivity to any of the recombinant allergens tested.

Increased Food Resources Help Eastern Oyster Mitigate the Negative Impacts of Coastal Acidification.

Oceanic absorption of atmospheric CO2 results in alterations of carbonate chemistry, a process coined ocean acidification (OA). The economically and ecologically important eastern oyster (Crassostrea virginica) is vulnerable to these changes because low pH hampers CaCO3 precipitation needed for shell formation. Organisms have a range of physiological mechanisms to cope with altered carbonate chemistry; however, these processes can be energetically expensive and necessitate energy reallocation. Here, the hypothesis that resilience to low pH is related to energy resources was tested. In laboratory experiments, oysters were reared or maintained at ambient (400 ppm) and elevated (1300 ppm) pCO2 levels during larval and adult stages, respectively, before the effect of acidification on metabolism was evaluated. Results showed that oysters exposed to elevated pCO2 had significantly greater respiration. Subsequent experiments evaluated if food abundance influences oyster response to elevated pCO2. Under high food and elevated pCO2 conditions, oysters had less mortality and grew larger, suggesting that food can offset adverse impacts of elevated pCO2, while low food exacerbates the negative effects. Results also demonstrated that OA induced an increase in oyster ability to select their food particles, likely representing an adaptive strategy to enhance energy gains. While oysters appeared to have mechanisms conferring resilience to elevated pCO2, these came at the cost of depleting energy stores, which can limit the available energy for other physiological processes. Taken together, these results show that resilience to OA is at least partially dependent on energy availability, and oysters can enhance their tolerance to adverse conditions under optimal feeding regimes.

Molecular Features Associated with Resilience to Ocean Acidification in the Northern Quahog, Mercenaria mercenaria.

The increasing concentration of CO2 in the atmosphere and resulting flux into the oceans will further exacerbate acidification already threatening coastal marine ecosystems. The subsequent alterations in carbonate chemistry can have deleterious impacts on many economically and ecologically important species including the northern quahog (Mercenaria mercenaria). The accelerated pace of these changes requires an understanding of how or if species and populations will be able to acclimate or adapt to such swift environmental alterations. Thus far, studies have primarily focused on the physiological effects of ocean acidification (OA) on M. mercenaria, including reductions in growth and survival. However, the molecular mechanisms of resilience to OA in this species remains unclear. Clam gametes were fertilized under normal pCO2 and reared under acidified (pH ~ 7.5, pCO2 ~ 1200 ppm) or control (pH ~ 7.9, pCO2 ~ 600 ppm) conditions before sampled at 2 days (larvae), 32 days (postsets), 5 and 10 months (juveniles) and submitted to RNA and DNA sequencing to evaluate alterations in gene expression and genetic variations. Results showed significant shift in gene expression profiles among clams reared in acidified conditions as compared to their respective controls. At 10 months of exposure, significant shifts in allele frequency of single nucleotide polymorphisms (SNPs) were identified. Both approaches highlighted genes coding for proteins related to shell formation, bicarbonate transport, cytoskeleton, immunity/stress, and metabolism, illustrating the role these pathways play in resilience to OA.

Combination of RNAseq and RADseq to Identify Physiological and Adaptive Responses to Acidification in the Eastern Oyster (Crassostrea virginica).

Ocean acidification (OA) is a major stressor threatening marine calcifiers, including the eastern oyster (Crassostrea virginica). In this paper, we provide insight into the molecular mechanisms associated with resilience to OA, with the dual intentions of probing both acclimation and adaptation potential in this species. C. virginica were spawned, and larvae were reared in control or acidified conditions immediately after fertilization. RNA samples were collected from larvae and juveniles, and DNA samples were collected from juveniles after undergoing OA-induced mortality and used to contrast gene expression (RNAseq) and SNP (ddRADseq) profiles from animals reared under both conditions. Results showed convergence of evidence from both approaches, particularly in genes involved in biomineralization that displayed significant changes in variant frequencies and gene expression levels among juveniles that survived acidification as compared to controls. Downregulated genes were related to immune processes, supporting previous studies demonstrating a reduction in immunity from exposure to OA. Acclimation to OA via regulation of gene expression might confer short-term resilience to immediate threats; however, the costs may not be sustainable, underscoring the importance of selection of resilient genotypes. Here, we identified SNPs associated with survival under OA conditions, suggesting that this commercially and ecologically important species might have the genetic variation needed for adaptation to future acidification. The identification of genetic features associated with OA resilience is a highly-needed step for the development of marker-assisted selection of oyster stocks for aquaculture and restoration activities.

Personalized Cancer Monitoring Assay for the Detection of ctDNA in Patients with Solid Tumors.

BACKGROUND: Highly sensitive molecular assays have been developed to detect plasma-based circulating tumor DNA (ctDNA), and emerging evidence suggests their clinical utility for monitoring minimal residual disease and recurrent disease, providing prognostic information, and monitoring therapy responses in patients with solid tumors. The Invitae Personalized Cancer Monitoring™ assay uses a patient-specific, tumor-informed variant signature identified through whole exome sequencing to detect ctDNA in peripheral blood of patients with solid tumors. METHODS: The assay's tumor whole exome sequencing and ctDNA detection components were analytically validated using 250 unique human specimens and nine commercial reference samples that generated 1349 whole exome sequencing and cell-free DNA (cfDNA)-derived libraries. A comparison of tumor and germline whole exome sequencing was used to identify patient-specific tumor variant signatures and generate patient-specific panels, followed by targeted next-generation sequencing of plasma-derived cfDNA using the patient-specific panels with anchored multiplex polymerase chain reaction chemistry leveraging unique molecular identifiers. RESULTS: Whole exome sequencing resulted in overall sensitivity of 99.8% and specificity of > 99.9%. Patient-specific panels were successfully designed for all 63 samples (100%) with ≥ 20% tumor content and 24 (80%) of 30 samples with ≥ 10% tumor content. Limit of blank studies using 30 histologically normal, formalin-fixed paraffin-embedded specimens resulted in 100% expected panel design failure. The ctDNA detection component demonstrated specificity of > 99.9% and sensitivity of 96.3% for a combination of 10 ng of cfDNA input, 0.008% allele frequency, 50 variants on the patient-specific panels, and a baseline threshold. Limit of detection ranged from 0.008% allele frequency when utilizing 60 ng of cfDNA input with 18-50 variants in the patient-specific panels (> 99.9% sensitivity) with a baseline threshold, to 0.05% allele frequency when using 10 ng of cfDNA input with an 18-variant panel with a monitoring threshold (> 99.9% sensitivity). CONCLUSIONS: The Invitae Personalized Cancer Monitoring assay, featuring a flexible patient-specific panel design with 18-50 variants, demonstrated high sensitivity and specificity for detecting ctDNA at variant allele frequencies as low as 0.008%. This assay may support patient prognostic stratification, provide real-time data on therapy responses, and enable early detection of residual/recurrent disease.

A Transcriptomic Analysis of Phenotypic Plasticity in Crassostrea virginica Larvae under Experimental Acidification.

Ocean acidification (OA) is a major threat to marine calcifiers, and little is known regarding acclimation to OA in bivalves. This study combined physiological assays with next-generation sequencing to assess the potential for recovery from and acclimation to OA in the eastern oyster (Crassostrea virginica) and identify molecular mechanisms associated with resilience. In a reciprocal transplant experiment, larvae transplanted from elevated pCO2 (~1400 ppm) to ambient pCO2 (~350 ppm) demonstrated significantly lower mortality and larger size post-transplant than oysters remaining under elevated pCO2 and had similar mortality compared to those remaining in ambient conditions. The recovery after transplantation to ambient conditions demonstrates the ability for larvae to rebound and suggests phenotypic plasticity and acclimation. Transcriptomic analysis supported this hypothesis as genes were differentially regulated under OA stress. Transcriptomic profiles of transplanted and non-transplanted larvae terminating in the same final pCO2 converged, further supporting the idea that acclimation underlies resilience. The functions of differentially expressed genes included cell differentiation, development, biomineralization, ion exchange, and immunity. Results suggest acclimation as a mode of resilience to OA. In addition, the identification of genes associated with resilience can serve as a valuable resource for the aquaculture industry, as these could enable marker-assisted selection of OA-resilient stocks.

Masticatory Myoelectric Side Modular Ratio Asymmetry during Maximal Biting in Women with and without Temporomandibular Disorders.

There is no consensus on the role of electromyographic analysis in detecting and characterizing the asymmetries of jaw muscle excitation in patients with temporomandibular disorders (TMD). To analyze the TMD patients (n = 72) in comparison with the healthy controls (n = 30), the surface electromyography (sEMG) of the temporalis anterior muscle (TA) and masseter muscle (M) was recorded while a maximal biting task was performed. The differences in the asymmetry of the relationship between the masseter muscles were assessed in a module to determine the sensitivity (Sn) of binomial logistic models, based on the dominance of the TA or the M muscle, in accurately predicting the presence of TMD. All assumptions were met, and comparisons between the groups showed significant differences for the TA muscle ratio (p = 0.007), but not for the M muscle ratio (p = 0.13). The left side was predominant over the right side in the TMD group for both the TA (p = 0.02) and M muscles (p = 0.001), while the non-TMD group had a higher frequency of the right side. Binary logistic regression showed a significant model (χ2 = 9.53; p = 0.002) for the TA muscle with Sn = 0.843. The model for the M muscle also showed significance (χ2 = 8.03; p = 0.005) with Sn = 0.837. The TMD patients showed an increased TA muscle ratio and asymmetry of left dominance, compared to the healthy subjects. Both of the binomial logistic models, based on muscle dominance TA or M, were moderately sensitive for predicting the presence of TMD.