On the reproducibility of extrusion-based bioprinting: round robin study on standardization in the field.

The outcome of three-dimensional (3D) bioprinting heavily depends, amongst others, on the interaction between the developed bioink, the printing process, and the printing equipment. However, if this interplay is ensured, bioprinting promises unmatched possibilities in the health care area. To pave the way for comparing newly developed biomaterials, clinical studies, and medical applications (i.e. printed organs, patient-specific tissues), there is a great need for standardization of manufacturing methods in order to enable technology transfers. Despite the importance of such standardization, there is currently a tremendous lack of empirical data that examines the reproducibility and robustness of production in more than one location at a time. In this work, we present data derived from a round robin test for extrusion-based 3D printing performance comprising 12 different academic laboratories throughout Germany and analyze the respective prints using automated image analysis (IA) in three independent academic groups. The fabrication of objects from polymer solutions was standardized as much as currently possible to allow studying the comparability of results from different laboratories. This study has led to the conclusion that current standardization conditions still leave room for the intervention of operators due to missing automation of the equipment. This affects significantly the reproducibility and comparability of bioprinting experiments in multiple laboratories. Nevertheless, automated IA proved to be a suitable methodology for quality assurance as three independently developed workflows achieved similar results. Moreover, the extracted data describing geometric features showed how the function of printers affects the quality of the printed object. A significant step toward standardization of the process was made as an infrastructure for distribution of material and methods, as well as for data transfer and storage was successfully established.

Non-Invasive Three-Dimensional Cell Analysis in Bioinks by Raman Imaging.

3D bioprinting is an emerging biofabrication strategy using bioinks, comprising cells and biocompatible materials, to produce functional tissue models. Despite progress in building increasingly complex objects, biological analyses in printed constructs remain challenging. Especially, methods that allow non-invasive and non-destructive evaluation of embedded cells are largely missing. Here, we implemented Raman imaging for molecular-sensitive investigations on bioprinted objects. Different aspects such as culture formats (2D, 3D-cast, and 3D-printed), cell types (endothelial cells and fibroblasts), and the selection of the biopolymer (alginate, alginate/nanofibrillated cellulose, alginate/gelatin) were considered and evaluated. Raman imaging allowed for marker-independent identification and localization of subcellular components against the surrounding biomaterial background. Furthermore, single-cell analysis of spectral signatures, performed by multivariate analysis, demonstrated discrimination between endothelial cells and fibroblasts and identified cellular features influenced by the bioprinting process. In summary, Raman imaging was successfully established to analyze cells in 3D culture in situ and evaluate them with regard to the localization of different cell types and their molecular phenotype as a valuable tool for quality control of bioprinted objects.

The Internal-External Locus of Control Short Scale-4 (IE-4): A comprehensive validation of the English-language adaptation.

The Internal-External Locus of Control Short Scale-4 (IE-4) measures two dimensions of the personality trait locus of control with two items each. IE-4 was originally developed and validated in German and later translated into English. In the present study, we assessed the psychometric properties (i.e., objectivity, reliability, validity) of the English-language IE-4, compared these psychometric properties with those of the German-language source version, and tested measurement invariance across both language versions. Using heterogeneous quota samples from the UK and Germany, we find that the English-language adaptation has satisfactory reliability and plausible correlations with 11 external variables (e.g., general self-efficacy, self-esteem, impulsive behavior, Emotional Stability), which are comparable with those of the German-language source version. Moreover, metric measurement invariance of the scale holds when comparing the UK and Germany, implying the comparability of correlations based on the latent factors across the two nations. As an ultra-short scale (completion time < 30 s), IE-4 lends itself particularly to the assessment of locus of control in survey contexts in which assessment time or questionnaire space are limited. It can be applied in a variety of research disciplines, such as psychology, sociology, or economics.

Adipose stem cell-derived extracellular matrix represents a promising biomaterial by inducing spontaneous formation of prevascular-like structures by mvECs.

Tissue constructs of physiologically relevant scale require a vascular system to maintain cell viability. However, in vitro vascularization of engineered tissues is still a major challenge. Successful approaches are based on a feeder layer (FL) to support vascularization. Here, we investigated whether the supporting effect on the self-assembled formation of prevascular-like structures by microvascular endothelial cells (mvECs) originates from the FL itself or from its extracellular matrix (ECM). Therefore, we compared the influence of ECM, either derived from adipose-derived stem cells (ASCs) or adipogenically differentiated ASCs, with the classical cell-based FL. All cell-derived ECM (cdECM) substrates enabled mvEC growth with high viability. Prevascular-like structures were visualized by immunofluorescence staining of endothelial surface protein CD31 and could be observed on all cdECM and FL substrates but not on control substrate collagen I. On adipogenically differentiated ECM, longer and higher branched structures could be found compared with stem cell cdECM. An increased concentration of proangiogenic factors was found in cdECM substrates and FL approaches compared with controls. Finally, the expression of proteins associated with tube formation (E-selectin and thrombomodulin) was confirmed. These results highlight cdECM as promising biomaterial for adipose tissue engineering by inducing the spontaneous formation of prevascular-like structures by mvECs.

Development of achievement goals and their relation to academic interest and achievement in adolescence: A review of the literature and two longitudinal studies.

Adolescence is important for the development of achievement motivation, including achievement goal pursuit. Longitudinal research is scarce on adolescents' goal development and its implications for academic outcomes. In our research, we first present a systematic review of findings on achievement goals in adolescence. Then we report 2 longitudinal studies with German adolescents in which we investigated the separate as well as joint development of achievement goals, interest, and achievement in the domain of mathematics. Study 1 comprised 745 students assessed in 4 waves in grades 5-7 (43% female; age MT1 = 10.66). Study 2 comprised 1,420 students assessed in 4 waves in grades 5-8 (47% female; age MT1 = 10.58). Students reported their mastery, performance-approach, and performance-avoidance goals and their interest in mathematics. Mathematics achievement was assessed by school grades (Studies 1 and 2) and standardized test scores (Study 1). Data were analyzed using latent growth curve, multiple process, and cross-lagged models. Findings for the 2 studies evidenced a substantial degree of consistency. All goals decreased and the decrease became smaller over time in most instances. Controlling for nonfocal goals and demographic variables (socioeconomic status, gender), multiple process models revealed that change in mastery and performance-approach goals was positively related or unrelated to change in interest and achievement, whereas change in performance-avoidance goals was negatively related or unrelated to change in interest and achievement. Cross-lagged models revealed that relations between the achievement goals on the one hand and interest and achievement on the other hand were reciprocal rather than unidirectional. (PsycINFO Database Record (c) 2020 APA, all rights reserved).

Maternal Weight, Weight Gain, and Metabolism are Associated with Changes in Fetal Heart Rate and Variability.

OBJECTIVE: Prepregnancy obesity and extensive weight gain can lead to diseases in the offspring later in life. The aim of this study was to evaluate the effect of anthropometric and metabolic factors on the fetal autonomic nervous system (ANS) in uncomplicated pregnancies. METHODS: A total of 184 pregnant women in the second or third trimester were included, and for 104 women, maternal insulin sensitivity (ISI) was determined. Fetal heart rate (HR) and heart rate variability (HRV) were determined by magnetic recording. Associations of maternal prepregnancy BMI, weight gain, and ISI with fetal HR and HRV were evaluated by ANCOVA, partial correlation, and mediation analysis. RESULTS: HR was increased and HRV decreased in fetuses of mothers with overweight or obesity in comparison to normal-weight mothers. Fetal HR was negatively correlated with maternal weight gain. Maternal prepregnancy BMI was positively correlated with fetal high frequency and was negatively correlated with low frequency and low/high frequency ratio. Maternal ISI showed a negative correlation with fetal HR. CONCLUSIONS: The results show that the fetal ANS is sensitive to alterations of prepregnancy BMI, weight changes, and glucose metabolism. These findings highlight the importance of the intrauterine environment on the developing ANS and the possible programming of obesity.

Implementation and evaluation of the water safety plan approach for buildings.

The World Health Organization (WHO) promotes water safety plans (WSPs) - a risk-based management approach - for premise plumbing systems in buildings to prevent deterioration of drinking-water quality. Experience with the implementation of WSPs in buildings were gathered within a pilot project in Germany. The project included an evaluation of the feasibility and advantages of WSPs by all stakeholders who share responsibility in drinking-water safety. While the feasibility of the concept was demonstrated for all buildings, benefits reported by building operators varied. The more technical standards were complied with before implementing WSP, the less pronounced were the resulting improvements. In most cases, WSPs yielded an increased system knowledge and awareness for drinking-water quality issues. WSPs also led to improved operation of the premise plumbing system and provided benefits for surveillance authorities. A survey among the European Network of Drinking-Water Regulators on the existing legal framework regarding drinking-water safety in buildings exhibited that countries are aware of the need to manage risks in buildings' installations, but experience with WSP is rare. Based on the successful implementation and the positive effects of WSPs on drinking-water quality, we recommend the establishment of legal frameworks that require WSPs for priority buildings whilst accounting for differing conditions in buildings and countries.

High-Ability Grouping: Benefits for Gifted Students' Achievement Development Without Costs in Academic Self-Concept.

Effects of full-time ability grouping on students' academic self-concept (ASC) and mathematics achievement were investigated in the first 3 years of secondary school (four waves of measurement; students' average age at first wave: 10.5 years). Students were primarily from middle and upper class families living in southern Germany. The study sample comprised 148 (60% male) students from 14 gifted classes and 148 (57% male) students from 25 regular classes (matched by propensity score matching). Data analyses involved multilevel and latent growth curve analyses. Findings revealed no evidence for contrast effects of class-average achievement or assimilation effects of class type on students' ASC. ASC remained stable over time. Students in gifted classes showed higher achievement gains than students in regular classes.

Impact of Intrauterine Growth Restriction on Cognitive and Motor Development at 2 Years of Age.

Intrauterine growth restriction (IUGR), which is already known to be a risk factor for pathological intrauterine development, perinatal mortality, and morbidity, is now also assumed to cause both physical and cognitive alterations in later child development. In the current study, effects of IUGR on infantile brain function were investigated during the fetal period and in a follow-up developmental assessment during early childhood. During the fetal period, visual and auditory event-related responses (VER and AER) were recorded using fetal magnetoencephalography (fMEG). VER latencies were analyzed in 73 fetuses (14 IUGR fetuses) while AER latencies were analyzed in 66 fetuses (11 IUGR fetuses). Bayley Scales of Infant Development, Second Edition (BSID-II) were used to assess the developmental status of the infants at the age of 24 months. The Mental Development Index (MDI) was available from 66 children (8 IUGR fetuses) and the Psychomotor Development Index (PDI) from 63 children (7 IUGR fetuses). Latencies to visual stimulation were more delayed in IUGR than in small for gestational age (SGA) or appropriate for gestational age (AGA) fetuses, albeit not to any significant extent (p = 0.282). The MDI in former IUGR infants was significantly lower (p = 0.044) than in former SGA and AGA infants. However, IUGR had no impact on PDI (p = 0.213). These findings support the hypothesis that IUGR may constitute a risk factor for neurodevelopmental delay. Further investigation of the possible underlying mechanisms, as well as continued long-term developmental research, is therefore necessary.

Effects of achievement differences for internal/external frame of reference model investigations: A test of robustness of findings over diverse student samples.

BACKGROUND: Achievement in math and achievement in verbal school subjects are more strongly correlated than the respective academic self-concepts. The internal/external frame of reference model (I/E model; Marsh, 1986, Am. Educ. Res. J., 23, 129) explains this finding by social and dimensional comparison processes. We investigated a key assumption of the model that dimensional comparisons mainly depend on the difference in achievement between subjects. We compared correlations between subject-specific self-concepts of groups of elementary and secondary school students with or without achievement differences in the respective subjects. AIMS: The main goals were (1) to show that effects of dimensional comparisons depend to a large degree on the existence of achievement differences between subjects, (2) to demonstrate the generalizability of findings over different grade levels and self-concept scales, and (3) to test a rarely used correlation comparison approach (CCA) for the investigation of I/E model assumptions. SAMPLES: We analysed eight German elementary and secondary school student samples (grades 3-8) from three independent studies (Ns 326-878). METHOD: Correlations between math and German self-concepts of students with identical grades in the respective subjects were compared with the correlation of self-concepts of students having different grades using Fisher's Z test for independent samples. RESULTS: In all samples, correlations between math self-concept and German self-concept were higher for students having identical grades than for students having different grades. Differences in median correlations had small effect sizes for elementary school students and moderate effect sizes for secondary school students. CONCLUSIONS: Findings generalized over grades and indicated a developmental aspect in self-concept formation. The CCA complements investigations within I/E-research.

Profile formation of academic self-concept in elementary school students in grades 1 to 4.

Academic self-concept (ASC) is comprised of individual perceptions of one's own academic ability. In a cross-sectional quasi-representative sample of 3,779 German elementary school children in grades 1 to 4, we investigated (a) the structure of ASC, (b) ASC profile formation, an aspect of differentiation that is reflected in lower correlations between domain-specific ASCs with increasing grade level, (c) the impact of (internal) dimensional comparisons of one's own ability in different school subjects for profile formation of ASC, and (d) the role played by differences in school grades between subjects for these dimensional comparisons. The nested Marsh/Shavelson model, with general ASC at the apex and math, writing, and reading ASC as specific factors nested under general ASC fitted the data at all grade levels. A first-order factor model with math, writing, reading, and general ASCs as correlated factors provided a good fit, too. ASC profile formation became apparent during the first two to three years of school. Dimensional comparisons across subjects contributed to ASC profile formation. School grades enhanced these comparisons, especially when achievement profiles were uneven. In part, findings depended on the assumed structural model of ASCs. Implications for further research are discussed with special regard to factors influencing and moderating dimensional comparisons.

Neuromagnetic signatures of syllable processing in fetuses and infants provide no evidence for habituation.

BACKGROUND: Habituation, as a basic form of learning, is characterized by decreasing amplitudes of neuronal reaction following repeated stimuli. Recent studies indicate that habituation to pure tones of different frequencies occurs in fetuses and infants. AIMS: Neural processing of different syllables in fetuses and infants was investigated. STUDY DESIGN: An auditory habituation paradigm including two different sequences of syllables was presented to each subject. Each sequence consisted of eight syllables (sequence /ba/: 5× /ba/, 1× /bi/ (dishabituator), 2× /ba/; sequence /bi/: 5× /bi/, 1× /ba/ (dishabituator), 2× /bi/). Each subject was stimulated with 140 sequences. Neuromagnetic signatures of auditory-evoked responses (AER) were recorded by fetal magnetoencephalography (fMEG). SUBJECTS: Magnetic brain signals of N=30 fetuses (age: 28-39weeks of gestation) and N=28 infants (age: 0-3months) were recorded. Forty-two of the 60 fetal recordings and 29 of the 58 infant recordings were included in the final analysis. OUTCOME MEASURES: AERs were recorded and amplitudes were normalized to the amplitude of the first stimulus. RESULTS: In both fetuses and infants, the amplitudes of AERs were found not to decrease with repeated stimulation. In infants, however, amplitude of syllable 6 (dishabituator) was significantly increased compared to syllable 5 (p=0.026). CONCLUSIONS: Fetuses and infants showed AERs to syllables. Unlike fetuses, infants showed a discriminative neural response to syllables. Habituation was not observed in either fetuses or infants. These findings could be important for the investigation of early cognitive competencies and may help to gain a better understanding of language acquisition during child development.

Big fish in big ponds: Contrast and assimilation effects on math and verbal self-concepts of students in within-school gifted tracks.

BACKGROUND: Ability grouping can lower students' academic self-perceptions through reference group effects of class-average achievement on academic self-concept (ASC) - the Big-fish-little-Pond-effect (BFLPE; Marsh & Parker, 1984, J. Pers. Soc. Psychol., 47, 213). Although the effect itself is well documented, many open questions remain. For instance, negative contrast effects of group average achievement and positive assimilation effects of group status are confounded in most BFLPE studies. For the verbal domain, no study has yet investigated contrast and assimilation effects simultaneously. Strong assimilation effects can be assumed for gifted ability grouping. AIMS: We aimed at disentangling contrast and assimilation effects of full-time within-school gifted tracking on verbal ASC and math ASC. SAMPLE: Students attended regular or gifted classes within the top track of German secondary schools. Our sample comprised 1,330 fifth-grade students (42 regular classes, n = 1,069, 48% female; 15 gifted classes, n = 261, 39% female). METHODS: Using multilevel regression analyses, we simultaneously modelled negative contrast effects of class ability and positive assimilation effects of class type (regular vs. gifted) on ASC in math and the verbal domain. RESULTS AND CONCLUSIONS: In the mathematical domain, the assimilation effect, caused by membership in gifted classes, compensated for negative contrast effects of class-average achievement on ASC even after controlling for previous ASC. In the verbal domain, we found neither a significant contrast effect nor an assimilation effect. Implications for gifted education are discussed.

Gestational Diabetes Impairs Human Fetal Postprandial Brain Activity.

CONTEXT: Gestational diabetes (GDM) influences the fetal phenotype. OBJECTIVE: In the present study, our aim was to determine the effect of GDM specifically on fetal brain activity. DESIGN: Pregnant participants underwent an oral glucose tolerance test (OGTT, 75 g). At 0, 60, and 120 minutes, maternal metabolism was determined, and fetal auditory evoked fields were recorded with a fetal magnetoencephalographic device. SETTING: All measurements were performed at the fMEG Center in Tübingen. PARTICIPANTS: Twelve women with GDM and 28 normal glucose-tolerant (NGT) pregnant women participated on a voluntary basis. INTERVENTIONS: OGTT (75 g, 120 minutes) was used in this study. MAIN OUTCOMES AND MEASURES: Fetal auditory evoked response latencies were determined for this study. RESULTS: In the fetuses of NGT women, latencies decreased between 0 and 60 minutes from 260 ± 90 to 206 ± 74 ms (P = .008) and remained stable until 120 minutes (206 ± 74 vs 230 ± 79, P =.129). In fetuses of women with GDM, there was no change in response latencies during OGTT (P = .11). Sixty minutes after glucose ingestion, fetal latencies in the GDM group were longer than in the NGT group (296 ± 82 vs 206 ± 74 ms, P = .001). Linear regression revealed a significant effect of maternal glucose, insulin levels, and insulin sensitivity on response latencies after 60 minutes. CONCLUSIONS: Fetal postprandial brain responses were slower in the offspring of women with GDM. This might indicate that gestational diabetes directly affects fetal brain development and may lead to central nervous insulin resistance in the fetus.

Heart rate variability parameters and fetal movement complement fetal behavioral states detection via magnetography to monitor neurovegetative development.

Fetal behavioral states are defined by fetal movement and heart rate variability (HRV). At 32 weeks of gestational age (GA) the distinction of four fetal behavioral states represented by combinations of quiet or active sleep or awakeness is possible. Prior to 32 weeks, only periods of fetal activity and quiesence can be distinguished. The increasing synchronization of fetal movement and HRV reflects the development of the autonomic nervous system (ANS) control. Fetal magnetocardiography (fMCG) detects fetal heart activity at high temporal resolution, enabling the calculation of HRV parameters. This study combined the criteria of fetal movement with the HRV analysis to complete the criteria for fetal state detection. HRV parameters were calculated including the standard deviation of the normal-to-normal R-R interval (SDNN), the mean square of successive differences of the R-R intervals (RMSSD, SDNN/RMSSD ratio, and permutation entropy (PE) to gain information about the developing influence of the ANS within each fetal state. In this study, 55 magnetocardiograms from healthy fetuses of 24-41 weeks' GA were recorded for up to 45 min using a fetal biomagnetometer. Fetal states were classified based on HRV and movement detection. HRV parameters were calculated for each state. Before GA 32 weeks, 58.4% quiescence and 41.6% activity cycles were observed. Later, 24% quiet sleep state (1F), 65.4% active sleep state (2F), and 10.6% active awake state (4F) were observed. SDNN increased over gestation. Changes of HRV parameters between the fetal behavioral states, especially between 1F and 4F, were statistically significant. Increasing fetal activity was confirmed by a decrease in PE complexity measures. The fHRV parameters support the differentiation between states and indicate the development of autonomous nervous control of heart rate function.

Magnetoencephalographic signatures of numerosity discrimination in fetuses and neonates.

Numerosity discrimination has been demonstrated in newborns, but not in fetuses. Fetal magnetoencephalography allows non-invasive investigation of neural responses in neonates and fetuses. During an oddball paradigm with auditory sequences differing in numerosity, evoked responses were recorded and mismatch responses were quantified as an indicator for auditory discrimination. Thirty pregnant women with healthy fetuses (last trimester) and 30 healthy term neonates participated. Fourteen adults were included as a control group. Based on measurements eligible for analysis, all adults, all neonates, and 74% of fetuses showed numerical mismatch responses. Numerosity discrimination appears to exist in the last trimester of pregnancy.

Maternal insulin sensitivity is associated with oral glucose-induced changes in fetal brain activity.

AIMS/HYPOTHESIS: Fetal programming plays an important role in the pathogenesis of type 2 diabetes. The aim of the present study was to investigate whether maternal metabolic changes during OGTT influence fetal brain activity. METHODS: Thirteen healthy pregnant women underwent an OGTT (75 g). Insulin sensitivity was determined by glucose and insulin measurements at 0, 60 and 120 min. At each time point, fetal auditory evoked fields were recorded with a fetal magnetoencephalographic device and response latencies were determined. RESULTS: Maternal insulin increased from a fasting level of 67 ± 25 pmol/l (mean ± SD) to 918 ± 492 pmol/l 60 min after glucose ingestion and glucose levels increased from 4.4 ± 0.3 to 7.4 ± 1.1 mmol/l. Over the same time period, fetal response latencies decreased from 297 ± 99 to 235 ± 84 ms (p = 0.01) and then remained stable until 120 min (235 ± 84 vs 251 ± 91 ms, p = 0.39). There was a negative correlation between maternal insulin sensitivity and fetal response latencies 60 min after glucose ingestion (r = 0.68, p = 0.02). After a median split of the group based on maternal insulin sensitivity, fetuses of insulin-resistant mothers showed a slower response to auditory stimuli (283 ± 79 ms) than those of insulin-sensitive mothers (178 ± 46 ms, p = 0.03). CONCLUSIONS/INTERPRETATION: Lower maternal insulin sensitivity is associated with slower fetal brain responses. These findings provide the first evidence of a direct effect of maternal metabolism on fetal brain activity and suggest that central insulin resistance may be programmed during fetal development.

Fetal behavioral states are stable over daytime - evidence by longitudinal and cross-sectional fetal biomagnetic recordings.

AIMS: Fetal behavioral states can be distinguished by biomagnetic recordings. We performed a longitudinal and a cross-sectional study to address the question whether the distribution of fetal behavioral states changes during the daytime. METHODS: For the longitudinal study, 32 magnetocardiographic recordings were performed on a singleton pregnancy on a weekly basis. On each examination day, two recordings were performed at different times between 25 and 40 weeks of gestation. For the cross-sectional study, fetal magnetocardiograms (fMCG) were recorded in a group of 32 singleton pregnancies matched for gestational age and daytime to the longitudinal study. The recordings were separated into two gestational age groups (less and more than 32 weeks). Fetal behavioral states were extracted from actocardiograms generated from MCG. RESULTS: No significant differences in fetal behavioral state distribution were found between morning and afternoon recordings in either the longitudinal or the cross-sectional study. CONCLUSION: This is the first magnetographic approach to show that daytime does not influence the distribution of fetal behavioral states in standardized recordings of 30 min length. This result implies that fetal magnetography recordings at normal daytimes can be combined without a bias and future recordings can be conducted independently of daytime as long as the varying behavioral states are generally taken into account during analysis.