Impact of temperature on physical and cognitive performance in elite female football players during intermittent exercise.

There is limited research on female football players, especially related to their physical and cognitive performance under different climactic conditions. We analyzed the impact of a hot environmental temperature on physical performance and anticipation in elite female football players during a fatigue-inducing intermittent protocol. Elite female players (n = 21) performed the countermovement jump (CMJ) and responded to filmed sequences of offensive play under two distinct environmental temperatures (i.e., mild environment temperature- 20°C and 30% rh versus hot environment temperature- 38°C and 80% rh), interspersed by 1-week interval. Linear mixed models were used. CMJ performance declined following the intermittent protocol on both temperature conditions (p < 0.05). Moreover, there were significant main effects for protocol on CMJ speed (m/s) (p = 0.001; ηp 2 = 0.12), CMJ power (p = 0.002; ηp 2 = 0.11), and CMJ Heightmax (p = 0.002; ηp 2 = 0.12). After performing the intermittent protocol, exposure to a hot temperature caused a greater decline in anticipation accuracy (mild temperature = 64.41% vs. hot temperature = 53.44%; p < 0.001). Our study shows impaired performance in elite female football players following an intermittent protocol under hot compared with mild environmental conditions. We report decreased performance in both CMJ and anticipation performance under hotter conditions. The results reveal that exposure to hot temperatures had a negative effect on the accuracy of their anticipatory behaviors. We consider the implication of the work for research and training interventions.

Endurance Performance Adaptations between SSG and HIIT in Soccer Players: A Meta-analysis.

The objective of this systematic review with meta-analysis was to compare the endurance performance chronic adaptations induced by running-based high-intensity interval training (HIIT), small-sided games (SSGs), and combined HIIT+SSGs in male and female youth and adult soccer players. The studies included in this review followed the PICOS criteria: (i) healthy soccer players; (ii) interventions based on SSGs; (iii) comparators exposed to only HIIT or combined SSGs+HIIT; (iv) endurance performance variables. Studies were searched for in the following databases: (i) PubMed; (ii) Scopus; (iii) SPORTDiscus; (iv) Web of Science. After conducting an initial database search that retrieved a total of 5,389 records, a thorough screening process resulted in the inclusion of 20 articles that met the eligibility criteria. Sixteen studies reported outcomes related to endurance performance measured through field-based tests, while five studies provided results from direct measurements of maximal oxygen uptake (VO2max). Results showed a non-significant small-magnitude favoring effect for the HIIT groups compared to the SSG groups (ES=0.37, p=0.074) for endurance, while a non-significant small-magnitude favoring SSGs was observed (ES=-0.20, p=0.303) for VO2max. Despite the very low certainty of evidence, the findings suggest similar effects induced by both SSG and HIIT on improving endurance performance and VO2max.

Contemporary practices of Portuguese and Brazilian soccer coaches in designing and applying small-sided games.

This descriptive study aimed to investigate the current practices of Portuguese and Brazilian soccer coaches in the design and implementation of small-sided games (SSGs) in soccer. A total of 187 male coaches participated in the online survey, consisting of 82 Portuguese and 105 Brazilian individuals. These coaches held various positions within the technical staff, with 63 serving as head coaches, 38 as assistant coaches, 38 as physical trainers, and 48 in other roles related to the technical staff. Additionally, the participants represented both youth (n = 102) and adult competitive levels (n = 59), along with some who were not currently associated with a specific group. The survey consisted of 32 questions divided into three main sections: (i) the timing of SSG application, (ii) the methods used for applying SSGs, and (iii) the reasons for applying SSGs. The Chi-square test revealed a statistically significant association between nationality and the frequency of SSGs used in training sessions during the pre-season (p = 0.039) and in-season (p < 0.001). Moreover, significant association between nationality and the time allocated to employing SSGs for targeting aerobic training (p < 0.001) was found. There was a significant association between nationality and the weekly frequency of SSGs use for targeting sprint training (p = 0.019). The Chi-square test identified significant associations between nationality and the use of SSGs for targeting technical training (p = 0.002), as well as for tactical training (p = 0.002). In summary, this study underscores that SSGs are primarily employed to enhance aerobic fitness, change of direction, technical skills, and tactical behaviors. Coaches generally favor employing SSGs two to three times a week, with typical sessions lasting between 16 to 30 minutes. Notably, the major discrepancies between nationalities lie in the importance assigned to the use of SSGs. However, in practice, the formats and objectives for implementing SSGs remain relatively similar.

Arbitrary absolute vs. individualized running speed thresholds in team sports: A scoping review with evidence gap map.

The aims of this scoping review were (i) to characterize the main methodological approaches to assessing individualized running speed thresholds in team sports players; (ii) to assess the use of traditional arbitrary (absolute) thresholds compared to individualized running speed thresholds in team sports players; (iii) to provide an evidence gap map (EGM) about the approaches and study designs employed in investigations in team sports and (iv) to provide directions for future research and practical applications for the strength and conditioning field. Methods studies were searched for in the following databases: (i) PubMed; (ii) Scopus; (iii) SPORTDiscus and (iv) Web of Science. The search was conducted on 15/07/2022. Risk of bias was assessed using the Risk of Bias Assessment Tool for Nonrandomized Studies (RoBANS). From 3,195 potentially relevant articles, 36 were eligible for inclusion in this review. Of the 36 included articles, 27 (75%) focused on the use of arbitrary and individualized running speed thresholds to describe the locomotor demands (e.g., high intensity running) of players. Thirty-four articles used individualized speed running thresholds based on physical fitness assessments (e.g., 40-m linear sprint) or physical performance (e.g., maximal acceleration). This scoping review supported the need for a greater focus to be placed on improving the methodological aspects of using individualized speed running thresholds in team sports. More than just creating alternatives to arbitrary thresholds, it is essential to increase the replicability of methodological conditions whilst ensuring that research comparing the most adequate measures and approaches to individualization takes into consideration the population and context of each study.

Effects of pitch size on soccer players' physiological, physical, technical, and tactical responses during small-sided games: a meta-analytical comparison.

One of the most often-used task constraints in designing small-sided games (SSGs) is the manipulation of pitch size to promote increases or decreases in the relative area per player. Such adjustments cause changes in the acute responses during SSGs. This systematic review with meta-analysis aimed to compare the effects of smaller vs. larger pitch sizes on soccer players' physiological, physical, technical, and tactical responses during SSGs. Comparisons between smaller and larger pitches were not considered based on a specific size, but also between using at least two dimensions in the same comparative study, aiming to understand differences between using smaller and larger (independently of the specific dimensions). The data sources utilized were PubMed, PsycINFO, Scielo, Scopus, SPORTDiscus, and Web of Science. The database search initially yielded 249 titles. From those, 41 articles were eligible for the systematic review and meta-analysis. Results revealed that, compared to smaller pitches, SSGs played on larger pitches induced greater values for heart rate (p < 0.001; ES = 0.50), rate of perceived exertion (p < 0.001; ES = 0.70), total distance (p < 0.001; ES = 1.95), high-speed running (p < 0.001; ES = 1.20), stretch index (p < 0.001; ES = 1.02) and surface area (p < 0.001; ES = 1.54). No significant differences were found between pitch size regarding the numbers of accelerations (p = 0.232; ES = 0.45), decelerations (p = 0.111; ES = 0.85), passes (p = 0.897; ES = 0.02), dribbles (p = 0.823; ES = -0.05), or positional centroid (p = 0.053; ES = 0.56). Larger pitch sizes can be implemented as a meaningful task constraint to increase the internal and external load experienced by soccer players during SSGs, as well as to increase the dispersion of players while acting together. These results were found independent of format and age group.

Jump rope training effects on health- and sport-related physical fitness in young participants: A systematic review with meta-analysis.

The aim of this systematic review with meta-analysis was to assess the available body of published peer-reviewed articles related on the effects of jump rope training (JRT) compared with active/passive controls on health- and sport-related physical fitness outcomes. Searches were conducted in three databases, including studies that satisfied the following criteria: i) healthy participants; ii) a JRT programprogramme; iii) active or traditional control group; iv) at least one measure related to health- and sport-related physical fitness; v) multi-arm trials. The random-effects model was used for the meta-analyses. Twenty-one moderate-high quality (i.e., PEDro scale) studies were meta-analysed, involving 1,021 participants (male, 50.4%). Eighteen studies included participants with a mean age <18 years old. The duration of the JRT interventions ranged from 6 to 40 weeks. Meta-analyses revealed improvements (i.e., p = 0.048 to <0.001; ES = 0.23-1.19; I2 = 0.0-76.9%) in resting heart rate, body mass index, fat mass, cardiorespiratory endurance, lower- and upper-body maximal strength, jumping, range of motion, and sprinting. No significant JRT effects were noted for systolic-diastolic blood pressure, waist-hip circumference, bone or lean mass, or muscle endurance. In conclusion, JRT, when compared to active and passive controls, provides a range of small-moderate benefits that span health- and sport-related physical fitness outcomes.

Algae application in civil construction: A review with focus on the potential uses of the pelagic Sargassum spp. biomass.

Pelagic Sargassum, usually found at the Sargasso Sea and the Western portion of the North Atlantic and Gulf of Mexico, has been detected in many new locations through the tropical Atlantic. The huge biomass found from the African coast to the Caribbean was called the Great Atlantic Sargassum Belt and is responsible for the stranding of tons of algae on coastal regions. Despite the environmental, social, and economic impacts, sargassum is a valuable source for multiple uses at the industry, such as alginates, cosmetics, recycled paper and bioplastics, fertilizers, and as raw material for civil construction. This work presents a systematic literature review on the use of algae at the civil construction sector, with a focus on the valorization of the pelagic Sargassum spp. biomass, by identifying the potential applications related to the use of other algal species. The review considered other genera of marine algae and marine angiosperms, resulting in a total of 31 selected articles. The marine grass Posidonia oceanica was the most used species, found in eight published papers, followed by the red alga Kappaphycus alvarezii with four studies. Two articles were available on the use of pelagic Sargassum spp. (S. fluitans and S.natans) for construction materials (adobe and pavement), with potential good results. The literature presented results from the use of marine algae and sea grasses for particleboards, polymeric and cemented composites, adobe, pavement, facades, and roofs. This article provides a state-of-the-art review of algal application in the civil construction sector and points out the main directions for the potentialities on the insertion of the Sargassum spp. biomass into the production chain of the sector.

The Effects of the Uncontrolled Use of Biomodulina T on the Severity of Severe Acute Respiratory Syndrome-Coronavirus 2 Infection in Older Cuban Adults: An Open Label Evaluation.

BACKGROUND: The ongoing coronavirus disease 2019 (COVID-19) pandemic is a worldwide public health threat. Millions of people are at risk and older adults are more susceptible to developing the most serious manifestations of the disease, in part because of the effects of ageing on the immune system. Biomodulina T is an injectable immune modulator that has been licensed for use in Cuba for many years. OBJECTIVE: An open-label, uncontrolled trial was conducted to investigate whether or not it might be useful to prevent or modulate the serious effects of severe acute respiratory syndrome coronavirus 2 infections in older Cuban adults before the availability of vaccinations. METHODS: From April 12 to August 31, 2021, 1239 adults aged 60 years and older, unvaccinated against COVID-19 were recruited from the José Luis Dubrocq polyclinics, to receive Biomodulina T, 1 intramuscular 3 mL dose weekly for 6 weeks. Each person was visited at home weekly to be administered Biomodulina T. Once daily patients were seen by a medical student to collect information on any possible adverse events related to the medication as well as any symptoms of COVID-19. The possible usefulness of the intervention and its potential adverse events were assessed based on the number of older adults who became infected with COVID-19, and the severity of any symptoms reported or noted both during the 6-week treatment period and during an additional 6-week posttreatment observation period. RESULTS: Sixteen patients were diagnosed with symptomatic COVID-19 during the intervention using a specific reverse transcription polymerase chain reaction test. One patient died because of COVID-19. The most common preexisting diagnoses in treated patients included high blood pressure in 64.8%, diabetes mellitus in 19.85%, and ischemic cardiopathy in 13.88%. Biomodulina T was well tolerated. Only infrequent, mild, transient, and self-limited adverse events were identified. Both the incidence of COVID-19 infections and the overall mortality rate were lower in the treated patients than what was observed in the untreated general population of this Cuban province during the same time period. CONCLUSIONS: Although further, confirmatory, double-blind, controlled clinical trials are needed, Biomodulina T injections were well tolerated, and the results of this open, uncontrolled study suggest that it may have been useful to decrease the incidence and severity of symptomatic COVID-19 infection in these older Cuban adults. (Curr Ther Res Clin Exp. 2022; 82:XXX-XXX) © 2022 Elsevier HS Journals, Inc.

Serum concentrations of folic acid and cobalamin and energy metabolism of ewes as a function of the energy density of the diet, peripartum period, and pregnancy toxemia.

The aim of this study was to determine the serum levels of folic acid, cobalamin, and indicators of energy metabolism during the peripartum period of ewes submitted to different diets and with or without pregnancy toxemia. Forty Santa Inês ewes were divided into three groups, a group of 20 animals fed only roughage-diet with low energy density (G1), a group of 10 ewes submitted to diet containing roughage and concentrate-diet with high energy density (G2), and a group of 10 ewes with diagnosis of pregnancy toxemia (PT) (G3). In the ewes of groups G1 and G2, blood samples were collected -30, -15, -7 days pre-partum, on the day of partum, and 7, 15, and 30 days postpartum; blood samples from G3 animals were collected during the veterinary clinic care. Significant variations were observed in the plasma concentration of glucose and serum folic acid, cobalamin, fructosamine, NEFA, and ß-hydroxybutyrate in the groups fed diets with different energy levels, with and without PT (P < 0.05). A positive correlation of folic acid and cobalamin was observed with plasma glucose and serum NEFA concentrations, in addition to negative correlation of glucose with NEFA and ß-hydroxybutyrate and positive correlation between the latter biomarkers of energy metabolism. The ewes diagnosed with PT showed higher levels of folic acid, NEFA, and ß-hydroxybutyrate (P < 0.05). Ewes fed diet containing roughage and concentrate present higher serum levels of folic acid, cobalamin, and glucose, in the pre-partum period and in the lactation period. The results obtained contribute to better understanding of the application of blood concentration tests for folic acid and cobalamin in the energy metabolism and in PT in ewes.

Variability of internal and external loads and technical/tactical outcomes during small-sided soccer games: a systematic review.

Small-sided games (SSGs) are widely used in soccer training. However, some of the typical outcomes related to human responses during these games (namely internal and external load) may vary between sessions for similar practice conditions. Thus, the study of intra- and inter-bout variability in response to SSGs is progressively growing. This systematic review aimed to (1) identify studies that have examined the intra- and inter-session bouts' variability levels regarding the internal and external load and technical/tactical outcomes during SSGs and (2) summarize the main evidence. A systematic review of PubMed, SPORTDiscus, Cochrane, and Web of Science databases was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. From the 486 studies initially identified, 24 were fully reviewed, and their outcome measures were extracted and analyzed. Sixteen studies analyzed internal load, 13 studies analyzed external load variables, six studies analyzed technical execution, and two studies analyzed tactical behavior. All studies included SSGs with a range number of players between 2 to 14 (1 vs. 1 to 7 vs. 7 SSGs). Internal load and low-speed external load variables presented a low variability, while high variations were reported regarding the technical execution and high-speed external loads.

Small-sided games in volleyball: A systematic review of the state of the art.

Studies on small-sided games (SSG) in team sports have increased in recent decades. However, the literature concerning this training strategy in volleyball is sparse. This study aims to summarize and analyse the scientific evidence on SSG in volleyball. For this purpose, electronic searches were conducted in August 2021 in PubMed, Scielo, ScienceDirect, Scopus, SPORTDiscus, and Web of Science databases. As result, a total of 22 studies (3 cross-sectional, 7 quasi-experimental, and 12 randomized controlled trial) that used SSG in volleyball were included in the qualitative synthesis after applying the eligibility criteria. Despite the few studies available for each outcome, our results suggest that the SSG can be used as a methodological resource for volleyball teaching and training of educational, recreational, and high-performance character. In conclusion, the use of SSG in volleyball is a pedagogical and training alternative with positive effects on populations with different levels of training (school and university students, recreational adult players, and athletes) considering instructional approaches, sport knowledge, participation in Physical Education classes, health markers, physical fitness, and physiological, psychological, and tactical-technical variables. However, more studies need to be carried out using SSG in volleyball in different contexts, with different manipulations and variables.

Antigen matching for transfusion support in Brazilian female patients with sickle cell disease to reduce RBC alloimmunization.

BACKGROUND: Red blood cell (RBC) alloimmunization is a complication of patients with sickle cell disease (SCD) and it has a greater impact on pregnancy, leading to a risk of hemolytic disease of the newborn and reducing blood availability for pregnant women. This study proposed to evaluate antigen matching transfusion protocols, aiming to reduce RBC alloimmunization in Brazilian female patients with SCD. METHODS: Samples from female patients with SCD (153) and self-declared Afro-Brazilian donors (307) were genotyped for RBC antigens and RH variants were investigated. The transfusion needs of patients during 1-year period and the number of compatible donors were assessed using three antigen-matching transfusion protocols: prophylactic CEK antigen-matched RBCs, prophylactic extended antigen-matched RBCs, and extended-matched red blood cells (RBCs) only for alloimmunized patients. In addition, RH molecular matching has been proposed for patients carrying variant RHCE. RESULTS: Provision of CEK antigen-matched donors would have been possible in 92.4% of transfusion events while provision of prophylactic extended antigen-matched RBCs would cover 88.7% of the transfusion events. Extended antigen matching for alloimmunized patients would be efficient in 99% of the cases. The presence of partial D in 10 patients increased the need of D-negative donors. Compatible donors could be enough for four of the five patients with altered RHCE genotypes in both alleles. CONCLUSION: In Brazilians, screening African descent donors allows the implementation of prophylactic CEK and extended antigen-matching transfusion protocols to female patients with SCD to reduce RBC alloimmunization; however, the supply of compatible blood can be impaired for patients with Rh variants.

The association between perceptual-cognitive processes and response time in decision making in young soccer players.

In soccer, it is relevant to understand the roles of Systems 1 (intuitive) and 2 (deliberative) in perceptual-cognitive processes and how they influence response time when making decisions. The aim of this study was to analyse how response time in decision making managed by Systems 1 and 2 is associated to the perceptual-cognitive processes of young soccer players. Ninety young soccer players participated. Perceptual-cognitive processes were assessed through visual search strategies, cognitive effort, and verbal reports. Participants wore a mobile-eye tracking system while viewing 11-a-side match play video-based soccer simulations. Response time in decision making was used to create two sub-groups: faster and slower decision-makers. Results indicated that players with faster response time in decision making employed more fixations of shorter duration, displayed less cognitive effort, as well as a greater number of thought processes associated with planning. These results reinforce that there are differences in the way of using the perceptive-cognitive processes from the priority system in the decision-making process. It is concluded that faster decision making, managed by System 1, implies greater ability to employ visual search strategies and to process information, thus enabling increased cognitive efficiency.

Skeletal Muscle-Adipose Tissue-Tumor Axis: Molecular Mechanisms Linking Exercise Training in Prostate Cancer.

Increased visceral adiposity may influence the development of prostate cancer (PCa) aggressive tumors and cancer mortality. White adipose tissue (WAT), usually referred to as periprostatic adipose tissue (PPAT), surrounds the prostatic gland and has emerged as a potential mediator of the tumor microenvironment. Exercise training (ET) induces several adaptations in both skeletal muscle and WAT. Some of these effects are mediated by ET-induced synthesis and secretion of several proteins, known as myo- and adipokines. Together, myokines and adipokines may act in an endocrine-like manner to favor communication between skeletal muscle and WAT, as they may work together to improve whole-body metabolic health. This crosstalk may constitute a potential mechanism by which ET exerts its beneficial role in the prevention and treatment of PCa-related disorders; however, this has not yet been explored. Therefore, we reviewed the current evidence on the effects of skeletal muscle-WAT-tumor crosstalk in PCa, and the potential mediators of this process to provide a better understanding of underlying ET-related mechanisms in cancer.

External and internal Load and their Effects on Professional Volleyball Training.

The purpose of this study was twofold: (i) characterize the external and internal training load of professional volleyball players with a focus on intra-week changes and (ii) test the relationships between internal and external load measures. Eight male professional players (age: 23.0±5.22 yo; body mass: 84.5 ± 7.58 kg; height: 193.0±9.71 cm; BMI: 22.0±0.02 kg/m2) were monitored daily over 15 weeks. The monitoring process included both internal (rate of perceived exertion [RPE] and session-RPE [s-RPE]) and external load variables, which were measured by an inertial measurement unit. Results revealed that, within-week variations revealed that RPE was significantly higher during MD-2 (d=0.59) and MD-3 (d=0.56) than MD-1. A significantly higher number of jumps was observed on MD-2 than MD-1 (d=0.69). Considering the relationships between internal and external load measures, small positive correlations were found between RPE and the number of jumps (r=0.17) and between s-RPE and the number of jumps (r=0.49). In conclusion, a tapering strategy was observed on the day before a match, as internal and external loads decreased. Both internal and external load measures are necessary to provide an accurate perception of the impact of training stimuli on players.

The Effects of Agility Ladders on Performance: A Systematic Review.

The goal of this systematic review was to evaluate the effects of exercise programs using agility ladders and to assess the quality of available evidence. Search was conducted in October of 2019 using the following databases: Cochrane Library, PEDro, PubMed, Scopus and Web of Science. Study eligibility criteria included randomized trials or randomized controlled trials using agility ladders drills. Participants were healthy humans of any health status. The study appraisal and synthesis methods followed the revised Cochrane risk-of-bias tool for randomized trials (RoB 2) and a qualitative synthesis of the main results of each study were applied. Only five studies met our criteria, lasting between 4 and 8 weeks. Only two studies evaluated the effects of ladder drills on more than one dimension. Lack of description of the specific exercises that were used limits reproducibility of current studies. With one exception, the articles had low risk of bias for most domains. Despite the widespread popularity of agility ladder drills, research is scarce and problematic, with poorly described protocols and mostly unidimensional performance measures. Claims that agility ladders improve agility and other physical skills is premature, given the nature and quality of existing research.

Accelerometry-based variables in professional soccer players: comparisons between periods of the season and playing positions.

The aim of this study was to provide reference data of variation in external training loads for weekly periods within the annual season. Specifically, we aimed to compare the weekly acute load, monotony, and training strain of accelerometry-based measures across a professional soccer season (pre-season, first and second halves of the season) according to players' positions. Nineteen professional players were monitored daily for 45 weeks using an 18-Hz global positioning system to obtain measures of high metabolic load distance (HMLD), impacts, and high intensity accelerations and decelerations. Workload indices of acute load, training monotony, and training strain were calculated weekly for each of the measures. The HMLD had greater training strain values in the pre-season than in the first (p ≤ 0.001; d = 0.793) and second halves of the season (p ≤ 0.001; d = 0.858). Comparisons between playing positions showed that midfielders had the highest weekly acute load of HMLD (6901 arbitrary units [AU]), while central defenders had the lowest (4986 AU). The pre-season period was associated with the highest acute and strain load of HMLD and number of impacts, with a progressive decrease seen during the season. In conclusion, coaches should consider paying greater attention to variations in HMLD and impacts between periods of the season and between players to individualize training accordingly.

Characterization of RHD alleles present in serologically RHD-negative donors determined by a sensitive microplate technique.

BACKGROUND AND OBJECTIVES: Weak D phenotypes with very low antigen densities and DEL phenotype may not be detected in RhD typing routine and could be typed as D-negative, leading to D alloimmunization of D-negative recipients. The present study aimed to investigate the presence of RHD-positive genotypes in blood donors typed as D-negative by an automated system using the solid-phase methodology as a confirmatory test. METHODS: Two screenings were performed in different selected donor populations. For the first screening, we selected 1403 blood donor samples typed as D-negative regardless of the CE status, and in the second screening, we selected 517 donor samples typed as D-negative C+ and/or E+. RhD typing was performed by microplate in an automated equipment (Neo-Immucor®), and the confirmatory test was performed by solid-phase technique using Capture R® technology. A multiplex PCR specific to RHD and RHDψ was performed in a pool of 6 DNA samples. Sequencing of RHD exons was performed in all RHD-positive samples, and a specific PCR was used to identify the D-CE(4-7)-D hybrid gene. RESULTS AND CONCLUSION: No weak D type was found in either screening populations. Additionally, 353 (18·4%) D-negative samples presented previously reported non-functional RHD genes, 2 samples had a DEL allele, and 6 samples demonstrated new alleles, including one novel DEL allele. Our study identified six new RHD alleles and showed that the inclusion of a confirmatory test using serological methodology with high sensitivity can reduce the frequency of weak D samples typed as D-negative.

Correlation among automated scores of agglutination, antigen density by flow cytometry and genetics of D variants.

BACKGROUND: Laboratory testing to identify the molecular basis of serologic weak D phenotypes is recommended to determine whether a pregnant woman or potential transfusion recipient should be managed as RhD-positive or RhD-negative. The variation in D antigen expression on RBCs, different potencies of anti-D typing reagents, lack of standardized test methods, and the subjectivity of interpreting agglutination reactions complicate the detection of D variants. We evaluated the correlation of agglutination scores by an automated immunoassay analyzer with D antigen densities determined by flow cytometry, and D variant types identified by molecular analysis. MATERIALS AND METHODS: We selected 273 blood donor samples with agglutination scores of less than 92 (4+), measured by an automated analyzer (NEO®, Immucor, Norcross, GA, USA). D antigen densities were measured by flow cytometry for 89 samples. Samples were classified as molecularly-determined weak D or partial D variants by multiplex PCR, PCR RFLP and DNA sequencing. RESULTS: All samples with a D antigen density ≥15% had an agglutination score >80 (4+). Agglutination scores for weak D types varied from 10 to 90. Agglutination scores for partial D antigens were graded with scores varying from 60 to 99. D antigen densities varied from 0.55% to 10.67% for weak Ds and 4.1% to 30.5% for partial Ds. DISCUSSION: Our results showed that score values follow a pattern among D variants that could be related to antigen density and to the RhD variant classification.