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In Flanders, open admission into higher education has led to heterogeneity in academic preparedness of incoming STEM students. Higher education institutions offer low stakes positioning tests to these students in order to help them assess their level of starting competences. Due to the unique nature of these tests, little can be inferred about students' perception of these tests. In this study, we used mixed methods to investigate these perceptions. Semi-structured interviews (n=20) and thematic analysis revealed that students do not always understand the stakes, objective and structure of positioning tests as intended by the organizers. A questionnaire (n=189) revealed that students considered chemistry problems better reflecting certain study programs than mathematics problems and that they wrongly think chemistry questions are more predictive than mathematics questions. Our findings provide insight in STEM students' multifaceted perceptions of and interaction with low stakes assessment.

The EGS Collab project performed well-monitored rock stimulation and flow tests at the 10-m scale in an underground research laboratory to inform challenges in implementing enhanced geothermal system (EGS) technology. This project, supported by the US Department of Energy, gathered data and observations from the field tests and compared these to simulation results to understand processes and to build confidence in numerical modeling of the processes. The project consisted of 3 Experiments, each comprising test and testbed design, many individual tests, numerical simulation, and analysis. The Experiments were performed in two deep underground testbeds at the Sanford Underground Research Facility (SURF) in Lead, South Dakota. Field experiments are now complete, significant data sets have been collected and analyzed, and some analysis continues. Experiments using underground test facilities have many advantages in that they allow:•Three-dimensional characterization of the stimulated volume by complementary geophysical methods surrounding the experiment•Using techniques that are currently not applicable under geothermal condition to provide processes insight •Comprehensive tracer testing and detailed characterization of complex fluid movements•Understanding the geometry of the stimulated network at the meso-scale and its implications for effective fracture surface area, rock block size, and heat exchange.Underground testing has its own set of complications, however, which affect the ability to perform tests as desired and affect the experiment results. Included here are the inability to flow at desired injection rates due to stress gradients caused by drift cooling, and the need to strongly limit induced seismicity because the distance to people and equipment.Experiment 1 examined hydraulic fracturing at a depth of 1.5 km in a well-characterized phyllite. Eight subhorizontal boreholes were used in this Experiment. Geophysical monitoring instrumentation was deployed in six boreholes to monitor stimulation events and flow tests. The other two boreholes were used to perform and carefully measure water injection and production. More than a dozen stimulations and nearly one year of flow tests in the testbed were performed. Detailed observations of processes occurring during stimulation and dynamic flow tests were collected and analyzed. Flow tests using ambient-temperature and chilled water were performed with intermittent tracer tests to examine system behavior. We achieved adaptive control of the tests using close monitoring of rapidly disseminated data and near-real-time simulation. Numerical simulation was critical in answering key experimental design questions, forecasting fracture behavior, and analyzing results. We were successful in performing many simulations in near-real-time in conjunction with the field experiments, with more detailed simulations performed later. Experiment 2 was intended to examine hydraulic shearing of natural fractures at a depth of 1.25 km in amphibolite. The stresses, rock type, and fracture conditions are different than in Experiment 1. The testbed consists of 9 boreholes, in addition to 2 exploratory characterization boreholes. Four boreholes drilled as two fans of 2 monitoring holes contained grouted-in monitoring sensors. The remaining five open boreholes drilled in a five-spot pattern were adaptively used for injection, production, and monitoring. Approximately five fracture set orientations were encountered in the testbed along with a low-stress rhyolite sill at 35 m below the access drift in exploratory well TV4100. The testbed was designed to optimize the potential for shear stimulation while also avoiding the low-stress rhyolite. Experiment 2 focused on stimulating a fracture in the most likely orientation to shear, however shear stimulation did not occur probably due to cementation from natural secondary mineralization. Other fracture sets encountered were also cemented and had orientations less likely to shear.Experiment 3 consisted of several stimulations in the same testbed as Experiment 2 allowing different stimulation approaches including ramped-rate injection, rapid injection, and oscillating-pressure injection. Ultimately these methods created hydraulic fractures, one of which was used for a medium-duration cold water injection test. The major findings of the EGS Collab Project include: 1.Significant shear stimulation did not occur during our stimulation attempts. Shear stimulation may occur but under a limited set of conditions not encountered.2.Our stimulations resulted in hydraulic fractures that required hydraulic propping. Pumping at pressures exceeding the minimum principal stress may not be feasible in an enhanced geothermal system.3.The systems we generated were complex hydraulic fracture/natural fracture systems, and these systems changed over time in response to applied pressures and flowrates and to unknown stimuli.4.The project attempted alternative stimulation methods, which did not provide significant flow improvement. 5.Thermal breakthrough was not achieved as designed, most likely because flow to production boreholes was not adequate. 6.The combination of geophysical tools used provided excellent understanding of many important processes. 7.Microearthquakes (MEQs) didn’t necessarily identify flow paths.8.Engineering tools bounding expected seismicity are needed.This report provides a summary of tests and analyses performed for EGS Collab Experiment 2 (Shear Stimulation in Testbed 2) and Experiment 3 (Alternative Stimulation methods). Much of the EGS Collab work has been published in journals and conference papers, presented in conferences, included in written reports, and submitted in data sets to the Geothermal Data Repository (GDR). The entirety of these written works is included as an appendix to this report, and this report serves as a summary and framework pointing to these published papers, presentations, and reports.

We conducted a time-lapse seismic experiment utilizing automated active seismic source and sensor arrays to monitor a reactivated fault within the Opalinus clay formation at the Mont Terri Rock Laboratory (Switzerland), an analog caprock for geologic carbon storage. A series of six brine injections were conducted into the so-called Main Fault to reactivate it. Seismic instrumentation in five monitoring boreholes on either side of the fault was used to continuously probe changes in P-wave travel-times associated with fault displacement and leakage. We performed time-lapse travel-time tomography on five hundred sequential data sets; this revealed a zone of decreased P-wave velocity, up to 16 m/s, during each injection cycle, followed by a velocity increase during shut-in. These results demonstrate varying elastic property perturbations, both spatially and temporally, along the fault plane during reactivation. We then interpreted these velocity changes in terms of fault dilation induced by pressurized fluids along the fault.

Abstract Background Microbial colonization during early life has a pivotal impact on the host health, shaping immune and metabolic functions, but little is known about timing and features of this process in dogs. The objectives of this study were to characterize the first step of intestinal microbiota development in naturally delivered canine puppies and to investigate its relationship with the maternal bacterial flora, using traditional culture and molecular analyses. Sixty puppies of two breeds, Appenzeller Cattle Dog (n = 3 dams) and Lagotto Romagnolo (n = 6), housed in the same breeding kennel, were included in the study. Swabs were collected in duplicate (for culture and for molecular analysis) from the dams’ vagina and rectum at the end of parturition, from puppies’ rectum, before maternal care, and from the environment (floor of the nursery and parturition box). Results 93.3% meconium samples showed bacterial growth, limited to a few colonies in 57.0% of cases. High growth was detected for Enterococcus faecalis, which was the most frequently isolated bacterium. The genus Enterococcus was one of the most represented in the dams’ rectum and vagina (88.9% and 55.6%, respectively). The genera Staphylococcus, Enterococcus, Escherichia and Proteus were also often isolated in meconium but were usually present in maternal samples as well, together with ubiquitous bacteria (Acinetobacter, Psychrobacter). In the environmental samples, just a few bacterial species were found, all with low microbial load. Additionally, bacteria of the phyla Proteobacteria, Firmicutes, and Actinobacteria were identified in meconium through molecular analysis, confirming the culture results and the early colonization of the newborn gut. Maternal, meconium and environmental samples had similar alpha diversity, while beta-diversity showed differences among families (i.e. a dam and her litter), and association indexes revealed a significant correlation between family members and between sample origin, suggesting a strong contribution of the maternal flora to the initial seeding of the canine neonatal gut and a strong individual dam imprint. Conclusion This study showed that the meconium of vaginally delivered puppies has its own microbiota immediately after birth, and that it is shaped by the dam, which gives a specific imprint to her litter.

Abstract Background Within the follicular fluid, extracellular vesicles (EVs) guide oocyte growth through their cargo microRNAs (miRNAs). Here, we investigated the role of EVs and their cargo miRNAs by linking the miRNAs found in EVs, derived from the fluid of an individual follicle, to the ability of its oocyte to become a blastocyst (competent) or not (non-competent). Methods Bovine antral follicles were dissected, categorized as small (2–4 mm) or large (5–8 mm) and the corresponding oocytes were subjected to individual maturation, fertilization and embryo culture to the blastocyst stage. Follicular fluid was pooled in 4 groups (4 replicates) based on follicle size and competence of the corresponding oocyte to produce a blastocyst. Follicular fluid-derived EVs were isolated, characterized, and subjected to miRNA-sequencing (Illumina Miseq) to assess differential expression (DE) in the 4 groups. Functional validation of the effect of miR-34c on embryo development was performed by supplementation of mimics and inhibitors during in vitro maturation (IVM). Results We identified 16 DE miRNAs linked to oocyte competence when follicular size was not considered. Within the large and small follicles, 46 DE miRNAs were driving blastocyst formation in each group. Comparison of EVs from competent small and large follicles revealed 90 DE miRNAs. Cell regulation, cell differentiation, cell cycle, and metabolic process regulation were the most enriched pathways targeted by the DE miRNAs from competent oocytes. We identified bta-miR-34c as the most abundant in follicular fluid containing competent oocytes. Supplementation of miR-34c mimic and inhibitor during IVM did not affect embryo development. However, blastocyst quality, as evidenced by higher cell numbers, was significantly improved following oocyte IVM in the presence of miR-34c mimics, while miR-34c inhibitors resulted in the opposite effect. Conclusion This study demonstrates the regulatory effect of miRNAs from follicular fluid-derived EVs on oocyte competence acquisition, providing a further basis for understanding the significance of miRNAs in oocyte maturation and embryonic development. Up-regulation of miR-34c in EVs from follicular fluid containing competent oocytes and the positive impact of miR-34c mimics added during IVM on the resulting blastocysts indicate its pivotal role in oocyte competence.

IntroductionSperm cryopreservation is a valuable technique for storing valuable canine genetics. However, little is known concerning the fate of frozen sperm stored in a sperm bank. This study aimed to characterize dogs presented for sperm cryopreservation and describe the use and popularity of frozen sperm in the Netherlands and Belgium over recent years.MethodsMedical records from dogs presented for sperm cryopreservation between January 1, 2014 and December 31, 2022 at two different freezing centers were reviewed retrospectively. Imported frozen sperm was excluded due to lack of usage information. Each sperm cryopreservation was considered a single event, and data were collected separately for each cryopreserved sample.ResultsA total of 3,090 ejaculates from 1,040 males of 157 different breeds were included and investigated using exploratory data analysis. The findings showed a steady rise in the popularity of sperm cryopreservation, with annual growth rates ranging from 8.4 to 41.9%. The majority of dogs (88.5%) were between 1 and 9 years old at the time of sperm cryopreservation, with nearly one-third aged 2–4 years. Most dogs were collected for sperm cryopreservation once (62.8%) or twice (21.6%). Sperm banks were used for both short- and long-term storage needs, and 6.83% of ejaculates were eventually discarded. The primary use of froze sperm was for international shipment, while 21.8% was used locally for artificial insemination. Depending on the year of cryopreservation, between 44.1 and 79.6% of frozen ejaculates remained unused or only partially used at the time of data collection.DiscussionThe results of this study provides the first comprehensive analysis of the use and popularity of frozen sperm among dog breeders in Belgium and the Netherlands and suggest a change in breeding practices in recent years. The percentage of breeders resorting to sperm cryopreservation and the extent of frozen sperm use in current breeding strategies remain to be defined in future studies.

Extracellular vesicles (EVs) contain microRNAs (miRNAs), which are important regulators of embryonic development. Nevertheless, little is known about the precise molecular processes controlling blastocyst development and quality. In a previous study, we identified bta-miR-665 as one of the miRNAs more abundantly present in extracellular vesicles of embryo-conditioned culture media of blastocysts compared to degenerate ones. Here, we investigated the effect and regulatory roles of bta-miR-665 in blastocyst development by supplementation of bta-miR-665 mimics or inhibitors to the culture media. Supplementation of bta-miR-665 mimics improved cleavage and blastocyst rate (P < 0.01), and blastocyst quality as indicated by increased inner cell mass rates and reduced apoptotic cell ratios (P < 0.01). Furthermore, supplementation of bta-miR-665 inhibitors had the opposite effect on these phenotypes. Low input transcriptome analysis and RT-qPCR revealed that bta-miR-665 acts on genes linked to microtubule formation and apoptosis/cell proliferation. These insights not only elucidate the important role of bta-miR-665 in embryo development, but also underscore its potential in improving reproductive efficiency in bovine embryo culture.

IntroductionThe increasing prevalence of infertility in male dogs in clinical practice mirrors current trends seen in human medicine. Acquired infertility is notably more common in dogs compared to congenital causes, with conditions such as testicular degeneration leading to irreversible loss of fertility. Current diagnostic methods for testicular degeneration, such as histopathological and cytological examinations, rely on testicular biopsy or fine needle aspiration, making them less feasible for routine use. Anti-Müllerian hormone (AMH), produced by Sertoli cells, has emerged as a potential alternative biomarker for testicular health, which can be measured in serum. This study evaluates AMH as a potential marker for testicular degeneration, using cryptorchid dogs as models for impaired fertility and altered testicular histology.MethodsThe relationship between serum AMH levels and AMH tissue expression with impaired spermatogenesis and altered histology was investigated. Serum AMH levels were determined in intact, cryptorchid, and castrated individuals using an immuno-enzymatic ELISA kit and compared between subgroups based on testicular location. Tissue AMH immuno-expression was differentially quantified in two regions of interest (ROIs), the interstitial space and the seminiferous tubule, in both descended and retained gonads. Furthermore, testicles were analyzed using histomorphometric analysis in seminiferous tubules, while spermatogenesis was evaluated using the Johnsen score.ResultsSerum AMH levels were positively correlated with AMH expression assessed in both interstitial space (ρ = 0.494, p ≤ 0.01) and seminiferous tubules (ρ = 0.610, p ≤ 0.001). Conversely, serum AMH levels showed a negative correlation with the seminiferous tubule area (ρ = −0.435, p ≤ 0.05). Smaller seminiferous tubule areas were linked to increased AMH reactivity in both seminiferous tubules (ρ = −0.774, p ≤ 0.001) and interstitial space (ρ = −0.725, p ≤ 0.001). Additionally, lower Johnsen scores were associated with higher serum AMH levels (ρ = −0.537, p ≤ 0.01) and elevated AMH expression in both seminiferous tubules (ρ = −0.756, p ≤ 0.001) and interstitial space (ρ = −0.679, p ≤ 0.001).DiscussionOur results suggest that higher serum levels and tissue expression of AMH are linked to smaller seminiferous tubules and poorer Johnsen scores, reflecting degenerative changes and Sertoli cell dysfunction in retained testicles. Given the similarities in the mechanisms that increase AMH levels in both cryptorchid and non-cryptorchid testicles affected by testicular degeneration, this study recommends using AMH as a marker for diagnosing testicular degeneration in dogs.

Abstract Maternal investment influences the survival and reproduction of both mothers and their progeny and plays a crucial role in understanding individuals’ life-history and population ecology. To reveal the complex mechanisms associated with reproduction and investment, it is necessary to examine variations in maternal investment across species. Comparisons across species call for a standardised method to quantify maternal investment, which remained to be developed. This paper addresses this limitation by introducing the maternal investment metric – MI – for mammalian species, established through the allometric scaling of the litter mass at weaning age by the adult mass and investment duration (i.e. gestation + lactation duration) of a species. Using a database encompassing hundreds of mammalian species, we show that the metric is not highly sensitive to the regression method used to fit the allometric relationship or to the proxy used for adult body mass. The comparison of the maternal investment metric between mammalian subclasses and orders reveals strong differences across taxa. For example, our metric confirms that Eutheria have a higher maternal investment than Metatheria. We discuss how further research could use the maternal investment metric as a valuable tool to understand variation in reproductive strategies.

Abstract Transfer RNA-derived small RNAs (tsRNAs) have been shown to be involved in early embryo development and repression of endogenous retroelements in embryos and stem cells. However, it is unknown whether tsRNAs also regulate embryo hatching. In this study, we mined the sequencing data of a previous experiment in which we demonstrated that the microRNA (miRNA) cargo of preimplantation embryonic extracellular vesicles (EVs) influences embryo development. We thus profiled the tsRNA cargo of EVs secreted by blastocysts and non-blastocysts. The majority of tsRNAs was identified as tRNA halves originating from the 5´ ends of tRNAs. Among the 148 differentially expressed tsRNAs, the 19 nt tRNA fragment (tRF) tDR-14:32-Glu-CTC-1 was found to be significantly up-regulated in EVs derived from non-blastocysts. RT-qPCR assays confirmed its significant up-regulation in non-blastocyst embryos and their conditioned medium compared to the blastocyst group (P

MicroRNAs (miRNA) are important regulators of oocyte maturation, playing a key role in modulating gene expression both in a temporal- and spatial-specific manner. These small non-coding RNAs are involved in important processes during oocyte maturation, acting as messengers between the oocyte and its surrounding cumulus cells. Despite its significance, the bidirectional communication mechanism is still unknown. To test miRNA communication between oocyte and surrounding cumulus cells through the gap junctions the gap junctions were either blocked with carbenoxolone or not. MiRNA sequencing of oocytes at 1, 6, and 22 h of in vitro maturation was then performed. Among the differentially expressed miRNAs, bta-miR-21-5p, a regulator of cumulus cell viability and oocyte maturation, was the only previously known miRNA. Furthermore, by labeling a bta-miR-21-5p mimic with FAM, crossing of this miRNA through the gap junctions within the cumulus-oocyte complex could be visualized and internalization in the oocyte was confirmed by RT-qPCR. In conclusion, this study provides, for the first time, evidence that miRNA communication within the bovine cumulus-oocyte complex is enabled through the gap junctional network.

Abstract Purpose This study aimed to investigate the difference in absolute and fat free mass (FFM)-adjusted resting energy expenditure (mREE) and body composition (body weight, fat mass (FM), FFM) between breast cancer survivors (BCs) and controls. Correlations with body composition were analyzed. We examined if survival year, or being metabolically dysfunctional were predictive variables. Methods A cross-sectional analysis was conducted on 32 BCs ≤5 years post treatment and 36 healthy controls. Indirect calorimetry measured absolute mREE. Body composition was determined by BOD POD. FFM-adjusted mREE was calculated (mREE/FFM). The Harris-Benedict equation was used to predict REE and determine hyper−/hypometabolism (mREE/pREE). The database of the multidisciplinary breast clinic of the University Hospital of Antwerp was consulted for survival year and metabolic dysfunctions. Results BCs have similar absolute mREE and greater FFM-adjusted mREE compared to controls. Absolute mREE and body composition between BCs differed; adjusted mREE was similar. FFM correlated significantly with absolute mREE in BCs. A significant interaction term was found between survival year and FM for absolute mREE. Conclusion BCs have similar absolute mREE, but higher FFM-adjusted mREE. Differences in body composition between BCs are suggested to cause inter-individual variations. We suggest that increased FFM-adjusted mREE is caused by metabolic stress related to cancer/treatment. Accurate measurement of REE and body composition is advised when adapting nutritional strategies, especially in patients at risk for developing metabolic dysfunctions.

Traffic monitoring systems featuring robust, multi-sensor fusion capabilities are rapidly growing in demand to observe traffic flow, reduce congestion and to detect and report traffic accidents. However, monitoring outdoor environments using cameras remains challenging due to complex weather conditions, including fog, rain, snow and variable lighting conditions. The presence of these weather conditions can significantly reduce vehicle detection and classification performance using machine learning methods. Unfortunately, openly available datasets for multi-sensor traffic monitoring development and testing remain limited, especially those featuring infrastructure-based cameras and millimeter wave (mmWave) radar. To address these challenges, we evaluate open camera and mmWave radar data using vehicle classification models for cars, trucks, vans and buses on embedded hardware. We also provide an open multi-sensor traffic monitoring dataset with more than 8,000 manually annotated frames as well as mmWave radar point clouds recorded in an urban environment under sunny, partially cloudy, cloudy, rainy and night conditions.

Abstract In different species, embryonic aneuploidies and genome-wide errors are a major cause of developmental failure. The increasing number of equine embryos being produced worldwide provides the opportunity to characterize and rank or select embryos based on their genetic profile prior to transfer. Here, we explored the possibility of generic, genome-wide preimplantation genetic testing concurrently for aneuploidies (PGT-A) and monogenic (PGT-M) traits and diseases in the horse, meanwhile assessing the incidence and spectrum of chromosomal and genome-wide errors in in vitro-produced equine embryos. To this end, over 70,000 single nucleotide polymorphism (SNP) positions were genotyped in 14 trophectoderm biopsies and corresponding biopsied blastocysts, and in 26 individual blastomeres from six arrested cleavage-stage embryos. Subsequently, concurrent genome-wide copy number detection and haplotyping by haplarithmisis was performed and the presence of aneuploidies and genome-wide errors and the inherited parental haplotypes for four common disease-associated genes with high carrier frequency in different horse breeds (GBE1, PLOD1, B3GALNT2, MUTYH), and for one color coat-associated gene (STX17) were compared in biopsy-blastocyst combinations. The euploid (n = 12) or fully aneuploid (n = 2) state and the inherited parental haplotypes for 42/45 loci of interest of the biopsied blastocysts were predicted by the biopsy samples in all successfully analyzed biopsy-blastocyst combinations (n = 9). Two biopsies showed a loss of maternal chromosome 28 and 31, respectively, which were confirmed in the corresponding blastocysts. In one of those biopsies, additional complex aneuploidies not present in the blastocyst were found. Five out of six arrested embryos contained chromosomal and/or genome-wide errors in most of their blastomeres, demonstrating their contribution to equine embryonic arrest in vitro. The application of the described PGT strategy would allow to select equine embryos devoid of genetic errors and pathogenetic variants, and with the variants of interest, which will improve foaling rate and horse quality. We believe this approach will be a gamechanger in horse breeding.

Abstract Background Nutritional support is often based on predicted resting energy expenditure (REE). In patients, predictions seem invalid. Indirect calorimetry is the gold standard for measuring EE. For assessments over longer periods (up to days), room calorimeters are used. Their design makes their use cumbersome, and warrants improvements to increase utility. Current study aims to compare data on momentary EE, obtained by a basic respiration room vs. classical ventilated hood. The objective is to compare results of the basic room and to determine its 1)reliability for measuring EE and 2)sensitivity for minute changes in activity. Methods Two protocols (P1; P2)(n = 62; 25 men/37 women) were applied. When measured by hood, participants in both protocols were in complete rest (supine position). When assessed by room, participants in P1 were instructed to stay half-seated while performing light desk work; in P2 participants were in complete rest mimicking hood conditions. The Omnical calorimeter operated both modalities. Following data were collected/calculated: Oxygen uptake ( $$\dot{\mathrm{V}}$$ V ˙ O2(ml/min)), carbon dioxide production ( $$\dot{\mathrm{V}}$$ V ˙ CO2ml/min), 24h_EE (kcal/min), and respiratory exchange ratio (RER). Statistical analyses were done between modalities and between protocols. The agreement between 24h_EE, $$\dot{\mathrm{V}}$$ V ˙ O2 and $$\dot{\mathrm{V}}$$ V ˙ CO2 obtained by both modalities was investigated by linear regression. Reliability analysis on 24h_EE determined ICC. Results No significant differences were found for 24h_EE and $$\dot{\mathrm{V}}$$ V ˙ O2. $$\dot{\mathrm{V}}$$ V ˙ CO2 significantly differed in P1 + P2, and P2 (hood > room). RER was significantly different (hood > room) for P1 + P2 and both protocols individually. Reliability of 24h_EE between modalities was high. Modality-specific results were not different between protocols. Discussion/conclusion The room is valid for assessing momentary EE. Minute changes in activity lead to a non-significant increase in EE and significant increase in RER. The significant difference in $$\dot{\mathrm{V}}$$ V ˙ CO2 for hood might be related to perceived comfort. More research is necessary on determinants of RER, type (intensity) of activity, and restlessness. The design of the room facilitates metabolic measurements in research, with promising results for future clinical use.

Abstract Background Breast cancer (BC) is the most prevalent tumor in women. Improvements in treatment led to declined mortality, resulting in more survivors living with cancer- or therapy-induced comorbidities. In this study, we investigated the impact of neoplasia and chemotherapy on resting energy expenditure (REE) and body composition, in relation to cancer-related fatigue. Inflammatory parameters were checked as possible explanation for changes in REE. Methods Fifty-six women participated: 20 women with BC and 36 healthy controls. Patients were assessed at baseline (T0) and follow-up (T1) after 12 weeks of chemotherapy. Controls were measured once. REE was assessed with indirect calorimetry: body composition (body weight, fat mass, fat-free mass) by air plethysmography. The multidimensional fatigue index (MFI-20) was used to analyze fatigue. Baseline measurements of patients were compared to results of the healthy controls with the independent-samples T-test. The paired-samples T-test investigated the effects of chemotherapy from T0 to T1. A Pearson correlation analysis was conducted between REE, body composition, and fatigue and between REE, body composition, and inflammatory parameters. A linear regression analysis was fitted to estimate the contribution of the significantly correlated parameters. The measured REE at T0 and T1 was compared to the predicted REE to analyze the clinical use of the latter. Results At baseline, patients with BC had significantly higher REE in the absence of differences in body composition. From baseline to T1, REE and body weight did not change. In contrast, fat-free mass declined significantly with concordant increase in fat mass. Fatigue deteriorated significantly. C-reactive protein at baseline predicted the change in energy expenditure. Predicted REE significantly underestimated measured REE. Conclusions Women with BC have higher REE in the tumor-bearing state compared to healthy controls. Chemotherapy does not affect REE but alters body composition. Predictive equations are invalid in the BC population. Results of our study can be used to implement personalized nutritional interventions to support energy expenditure and body composition and minimize long-term comorbidities.

The EGS Collab project is performing well-monitored rock stimulation and flow tests at the 10-m scale in an underground research laboratory to inform challenges in implementing enhanced geothermal systems (EGS). This project, supported by the US Department of Energy, is gathering data and observations from the field tests and comparing these to simulation results to understand processes and to build confidence in numerical modeling of the processes. Experiment 1 (now complete) examined hydraulic fracturing in an underground test bed at the Sanford Underground Research Facility (SURF) in Lead, South Dakota, at a depth of approximately 1.5 km in a well-characterized phyllite. Geophysical monitoring instrumentation in six of eight sub-horizontal boreholes monitored stimulation events and flow tests. The other two boreholes were used to perform and carefully measure water injection and production. More than a dozen stimulations and nearly one year of flow tests in the testbed were performed. Detailed observations of processes occurring during stimulation and dynamic flow tests were collected and analyzed. Flow tests using ambient-temperature and chilled water were performed with intermittent tracer tests to examine system behavior. We achieved adaptive control of the tests using close monitoring of rapidly disseminated data and near-real-time simulation. Numerical simulation was critical in answering key experimental design questions, forecasting fracture behavior, and analyzing results. We were successful in performing many simulations in near-real-time in conjunction with the field experiments, with more detailed simulations performed later. The primary objective of Experiment 2 is to examine hydraulic shearing of natural fractures at a depth of 1.25 km in amphibolite at SURF. The stresses, rock type, and fracture conditions are different than in Experiment 1. The testbed consists of 9 boreholes, in addition to two earlier-drilled characterization boreholes. One borehole is used for injection, two fans of 2 monitoring wells have several geophysical measurement tools grouted in, and four open boreholes surrounding the injection hole are adaptively used for production and monitoring. We have encountered approximately five fracture set orientations in the testbed, and designed our testbed accordingly to maximize the potential for shear stimulation. Three stimulations have been performed to date from the injection borehole, each intersecting at least one production borehole. Different methods have been used for each stimulation, including a ramped flow, a high flow rate, and oscillating pressure.

Abstract Epididymal spermatozoa obtained post mortem are considered a valuable source of genetic material which is often irrevocably lost. This makes these gametes constitute a key element in protection and restitution programs. The wisent (Bison bonasus, Linnaeus 1758) is a species that survived in zoos after extinction from its natural habitat. This resulted in a narrowing of the genetic pool of the whole population, which is at present derived from only 12 ancestors. Currently, wisent protection programs are aimed at preserving the genetic diversity by establishing a germplasm bank. The objective of this study was to comprehensively characterize the morphology, morphometry and functionality of wisent epididymal spermatozoa and evaluate the effectiveness of their cryopreservation in extender based on Tris buffer and chicken egg yolk. The median total number of spermatozoa obtained from one individual was 1985.0 × 106 (62.5 × 106–7452.0 × 106). These gametes were characterized by median: 40.0% (0.5–70.0%) subjective motility, 69.8% (32.5–90.0%) viability and 54.3% (10.5–83.3%) normal morphology. The sperm head had a median size of 5.0 μm (3.5–6.7 μm) width, 8.5 μm (6.4–11.3 μm) length and 36.9 μm2 (23.7–48.6 μm2) surface area. The viable population of the obtained gametes was characterized by median values 53.2% (4.5–80.3%) of intact sperm membrane, 50.8 (26.0–76.6%) of intact acrosome, 0.4% (0–98.7%) of fragmented chromatin, 5.9% (0.0–88.8%) of cells with high mitochondrial potential and 42.1% (8.3–63.7%) without lipid peroxidation. The viable population of the frozen/thawed gametes was characterized by median values: 18.4% (2.4–57.9%) of intact sperm membrane, 35.1 (11.9–56.7%) of intact acrosome, 0.07% (0–89.2%) of fragmented chromatin, 12.8% (0.0–49.7%) of cells with high mitochondrial potential and 16.3% (2.2–53.6%) without lipid peroxidation. Due to the material originating from a relatively large number of wild individuals, the research presented here contributed to the description of certain species standards for the assessment of wisent epididymal spermatozoa. The presented effect of cryopreservation on these gametes justifies the use of an extender based on Tris buffer with the addition of chicken egg yolk. The obtained effects are satisfactory from the point of view of preserving valuable genetic material and their use in ART.

Abstract We conducted a time‐lapse seismic experiment utilizing automated active seismic source and sensor arrays to monitor a reactivated fault within the Opalinus clay formation at the Mont Terri Rock Laboratory (Switzerland), an analog caprock for geologic carbon storage. A series of six brine injections were conducted into the so‐called Main Fault to reactivate it. Seismic instrumentation in five monitoring boreholes on either side of the fault was used to continuously probe changes in P‐wave travel‐times associated with fault displacement and leakage. We performed time‐lapse travel‐time tomography on five hundred sequential data sets; this revealed a zone of decreased P‐wave velocity, up to 16 m/s, during each injection cycle, followed by a velocity increase during shut‐in. These results demonstrate varying elastic property perturbations, both spatially and temporally, along the fault plane during reactivation. We then interpreted these velocity changes in terms of fault dilation induced by pressurized fluids along the fault.

Understanding the interactions among agricultural processes, soil, and plants is necessary for optimizing crop yield and productivity. This study focuses on developing effective monitoring and analysis methodologies that estimate key soil and plant properties. These methodologies include data acquisition and processing approaches that use unmanned aerial vehicles (UAVs) and surface geophysical techniques. In particular, we applied these approaches to a soybean farm in Arkansas to characterize the soil-plant coupled spatial and temporal heterogeneity, as well as to identify key environmental factors that influence plant growth and yield. UAV-based multitemporal acquisition of high-resolution RGB (red-green-blue) imagery and direct measurements were used to monitor plant height and photosynthetic activity. We present an algorithm that efficiently exploits the high-resolution UAV images to estimate plant spatial abundance and plant vigor throughout the growing season. Such plant characterization is extremely important for the identification of anomalous areas, providing easily interpretable information that can be used to guide near-real-time farming decisions. Additionally, high-resolution multitemporal surface geophysical measurements of apparent soil electrical conductivity were used to estimate the spatial heterogeneity of soil texture. By integrating the multiscale multitype soil and plant datasets, we identified the spatiotemporal co-variance between soil properties and plant development and yield. Our novel approach for early season monitoring of plant spatial abundance identified areas of low productivity controlled by soil clay content, while temporal analysis of geophysical data showed the impact of soil moisture and irrigation practice (controlled by topography) on plant dynamics. Our study demonstrates the effective coupling of UAV data products with geophysical data to extract critical information for farm management.