Standardization in the Diagnosis of Mixed Phenotype Acute Leukemia (MPAL): Semiquantitative, Universally Applicable Flow Cytometric Criteria for Immunophenotypic Lineage Assignment and Isolated MPO

Mixed phenotype acute leukemia (MPAL) is a rare group of acute leukemias defined by immunophenotypic expression of more than one hematopoietic cell lineage. The World Health Organization (WHO) diagnostic immunophenotypic criteria for MPAL rely on the intensity of lineage-defining antigen expression, predominantly a qualitative assessment, and are often difficult to apply to a phenotypically heterogeneous leukemia. Cases of MPAL defined by isolated myeloperoxidase (isoMPO) expression on otherwise typical acute lymphoblastic leukemia (ALL) are variably diagnosed as MPAL or ALL based on the incompletely defined criteria for assigning MPO expression. We hypothesized that quantitative criteria for antigen intensity could be developed and applied in a uniform manner across flow cytometry instruments, reagents, and analysis software to enable a consistent approach to diagnosing MPAL and better defining isoMPO. We previously reported on a multicenter cohort identified by respective institutions as MPAL with subsequent central review according to 2008 WHO criteria (Oberley et al 2020). Of these, 100 had suitable dot plots for reevaluation (89: B/Myeloid, 10: T/Myeloid, 1: B/T). Antigen expression was concurrently reviewed by two hematopathologists to reach consensus (BLW, AEK). The cohort was divided a priori into randomly selected training and testing cases (n=30/n=70). Classification criteria were generated in the training cohort for WHO lineage-defining antigen expression (myeloid: cMPO, CD64, CD14; B: CD19, T: cCD3) from 10 cases and then refined through review of an additional 20 cases. Positive antigen expression was classified as maximal intensity approaching that of the mature normal counterpart population (NCP) (cMPO: neutrophils; CD64, CD14 and CD11c: monocytes; CD19: B cells; cCD3: T cells) either by 1) range of expression recapitulating maturation of the NCP, irrespective of the percentage on the leukemic population (Figure 1A); or 2) uniform expression above background on a discrete (sub)population (Figure 1B). To account for technical variation within and among laboratories, maximal antigen intensity on the leukemic population was measured in 0.5 log increments and normalized to the maximal intensity of the NCP. An intensity of ≥50% of the NCP above background was defined as sufficient for MPAL lineage assignment and <50% consistent with isoMPO. This approach was then used to classify the remaining 70 cases. Using this approach, 41/98 (42%) cases previously classified as MPAL remained classified as MPAL: 31 as B/Myeloid (25 by maturational MPO expression, 6 by maturational CD64 and/or CD14 expression); 9 as T/Myeloid (8 by maturational MPO expression, 1 with maturational CD64, CD14 and CD11c expression); and 1 as B/T. No cases in the cohort showed uniform expression ≥50% of the NCP. The remaining 57 showed isolated low-level MPO expression (maximal intensity <50% of the NPC) on a uniform leukemic population or on a dichotomous subpopulation (isoMPO), 56 B/Myeloid and 1 T/Myeloid. Two cases of otherwise typical B-ALL without myeloid or monocytic antigen expression were reclassified as B-ALL, one of which showed low variable CD10 suggestive of DUX4-rearranged B-ALL. In comparison to our previously reported study of 2008 WHO-defined MPAL, 53/89 (60%) centrally-confirmed cases would be classified here as isoMPO. Conversely, five cases excluded under 2008 WHO central review would be reclassified as having sufficient antigen expression to qualify as MPAL (2 B/Myeloid, 3 T/Myeloid). Novel semiquantitative immunophenotypic criteria applied to a large cohort of acute leukemias originally diagnosed as MPAL was able to objectively identify a large subset as having dim, uniform MPO expression (isoMPO). Our approach emphasizes antigen expression recapitulating maturational expression similar to their non-leukemic cellular counterparts, normalizes absolute intensities to internal positive and negative control populations to minimize technical variability between observers and assays, and as per the 2017 WHO, does not require a specific percent threshold of positivity. While requiring validation, this is a critical first step toward establishing a reproducible delineation of these complex cases to practically implement the WHO classification to support treatment decisions and ongoing investigation into MPAL genomics and outcomes (available for this cohort by ASH). Figure 1 Figure 1. Disclosures Oberley: Caris LIfe Science: Current Employment. Orgel: Jazz Pharmaceuticals: Consultancy.

The Effects of High-Intensity Interval Exercise on Skeletal Muscle and Cerebral Oxygenation during Cycling and Isokinetic Concentric and Eccentric Exercise

The aim of the present study was to study the effects of cycling and pure concentric and pure eccentric high-intensity interval exercise (HIIE) on skeletal muscle (i.e., vastus lateralis) and cerebral oxygenation. Twelve healthy males (n = 12, age 26 ± 1 yr, body mass 78 ± 2 kg, height 176 ± 2 cm, body fat 17 ± 1% of body mass) performed, in a random order, cycling exercise and isokinetic concentric and eccentric exercise. The isokinetic exercises were performed on each randomly selected leg. The muscle and the cerebral oxygenation were assessed by measuring oxyhemoglobin, deoxyhemoglobin, total hemoglobin, and tissue saturation index. During the cycling exercise, participants performed seven sets of seven seconds maximal intensity using a load equal to 7.5% of their body mass while, during isokinetic concentric and eccentric exercise, they were performed seven sets of five maximal muscle contractions. In all conditions, a 15 s rest was adopted between sets. The cycling HIIE caused greater fatigue (i.e., greater decline in fatigue index) compared to pure concentric and pure eccentric isokinetic exercise. Muscle oxygenation was significantly reduced during HIIE in the three exercise modes, with no difference between them. Cerebral oxygenation was affected only marginally during cycling exercise, while no difference was observed between conditions. It is concluded that a greater volume of either concentric or eccentric isokinetic maximal intensity exercise is needed to cause exhaustion which, in turn, may cause greater alterations in skeletal muscle and cerebral oxygenation.

Monitoring Master Swimmers’ Performance and Active Drag Evolution along a Training Mesocycle

This study aimed to analyze the effects of a swimming training mesocycle in master swimmers’ performance and active drag. Twenty-two 39.87 ± 6.10 year-old master swimmers performed a 25 m front crawl at maximal intensity before and after a typical four-week training mesocycle. Maximum, mean and minimum speeds, speed decrease and hip horizontal intra-cyclic velocity variation were assessed using an electromechanical speedometer, and the active drag and power to overcome drag were determined using the measuring active drag system. Maximum, mean and minimum front crawl speeds improved from pre- to post-training (mean ± 95% CI: 3.1 ± 2.8%, p = 0.04; 2.9 ± 1.6%, p = 0.01; and 4.6 ± 3.1%, p = 0.01; respectively) and the speed decrease along the 25 m test lowered after the training period (82.5 ± 76.3%, p = 0.01). The training mesocycle caused a reduction in the active drag at speeds corresponding to 70% (5.0 ± 3.9%), 80% (5.6 ± 4.0%), and 90% (5.9 ± 4.0%), but not at 100% (5.9 ± 6.7%), of the swimmers’ maximal exertions in the 25 m test. These results showed that four weeks of predominantly aerobic training could improve master swimmers’ performance and reduce their hydrodynamic drag while swimming mainly at submaximal speeds.

RESEARCH OF EMISSIONS TO THE ATMOSPHERE FROM AUTOMOBILE TRANSPORT AND OF PECULIARITIES OF DISTRIBUTION OF AUTOMOBILE TRANSPORT FLOWS ON THE TERRITORY OF CITY DISTRICT TOGLIATTI OF RUSSIA

Complex of researches of emissions to the atmosphere causing by automobile transport on the territory of city district Togliatti was carried out. Measurements were carries out by universal portable gas analyzer “GANK-4”. Results of measurements are showing a number of exceeding of admissible norms. It was concluded that atmosphere air near to automobile roads of city district Togliatti with maximal intensity of movement of automobile transportis maximally polluted by carbon monoxide in automobile transport emissions. For determination of volume of emissions of automobile transport on city district Togliatti automobile roads and for further it using as source data during calculations of atmosphere pollutions study of peculiarities of distribution of automobile transport flows (its components and intensity) on the territory of city district Togliatti and of it variations in time was done. In total pollution of the territory of city district Togliatti by emissions causing by automobile transport, may be considered as serious ecological problem. It is necessary to develop and to implement the measures of it complex reduction.

Player Session Rating of Perceived Exertion: A More Valid Tool Than Coaches’ Ratings to Monitor Internal Training Load in Elite Youth Female Basketball

Purpose: To monitor elite youth female basketball training to verify whether players’ and coaches’ (3 technical coaches and 1 physical trainer) session rating of perceived exertion (s-RPE) has a relationship with Edwards’ method. Methods: Heart rate of 15 elite youth female basketball players (age 16.7 [0.5] y, height 178 [9] cm, body mass 72 [9] kg, body mass index 22.9 [2.2] kg·m−2) was monitored during 19 team (268 individual) training sessions (102 [15] min). Mixed effect models were applied to evaluate whether s-RPE values were significantly (P ≤ .05) related to Edwards’ data, total session duration, maximal intensity (session duration at 90–100% HRmax), type of training (ie, strength, conditioning, and technique), and whether differences emerged between players’ and coaches’ s-RPE values. Results: The results showed that there is a relationship between s-RPE and Edwards’ methods for the players’ RPE scores (P = .019) but not for those of the trainers. In addition, as expected, both players’ (P = .014) and coaches’ (P = .002) s-RPE scores were influenced by total session duration but not by maximal intensity and type of training. In addition, players’ and coaches’ s-RPE values differed (P < .001)—post hoc differences emerged for conditioning (P = .01) and technique (P < .001) sessions. Conclusions: Elite youth female basketball players are better able to quantify the internal training load of their sessions than their coaches, strengthening the validity of s-RPE as a tool to monitor training in team sports.