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The immune system plays a crucial role in tissue repair and regeneration. Macrophages have been identified as master regulators of the early immune response and healing outcome, by orchestrating the temporal nature of the initial inflammation phase and coordinating the fate of stem/progenitor cells involved in regeneration. However, traditional in-vitro models for the study of macrophages often fail to fully replicate the complexity of the in-vivo microenvironment, therefore generating models which do not fully capture the extensive spectrum of macrophage behaviour seen in native tissues. To this end, we used a hematoma-mimetic 3D fibrin matrix characteristic of early injured tissues to generate a 3D in-vitro model mirroring the local macrophage microenvironment. Leveraging this framework, we demonstrated significant effects of extracellular matrix and dimensionality on macrophage basal signalling and polarisation, achieving more pronounced regenerative phenotypes upon stimulation with the M2a polarisation factors compared to traditional 2D tissue culture conditions. Moreover, this enhanced physiological macrophage behaviour corresponded to increased coordination of angiogenesis and osteogenesis, better mirroring the healing processes seen in-vivo. Taken together, this study demonstrates the critical importance of integrating tissue composition and 3D architecture when investigating the macrophage behaviour in-vitro, establishing a powerful tool that overcomes known limitations associated with traditional 2D culture on plastic, and can be used to identify and validate novel immunomodulation strategies to enhance tissue regeneration., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Inc. All rights reserved.)

Motion remains a significant technical hurdle in fMRI studies of young children. Our aim was to develop a straightforward and effective method for obtaining and preprocessing resting state data from a high-motion pediatric cohort. This approach combines real-time monitoring of head motion with a preprocessing pipeline that uses volume censoring and concatenation alongside independent component analysis based denoising. We evaluated this method using a sample of 108 first grade children (age 6-8) enrolled in a longitudinal study of math development. Data quality was assessed by analyzing the correlation between participant head motion and two key metrics for resting state data, temporal signal-to-noise and functional connectivity. These correlations should be minimal in the absence of noise-related artifacts. We compared these data quality indicators using several censoring thresholds to determine the necessary degree of censoring. Volume censoring was highly effective at removing motion-corrupted volumes and ICA denoising removed much of the remaining motion artifact. With the censoring threshold set to exclude volumes that exceeded a framewise displacement of 0.3 mm, preprocessed data met rigorous standards for data quality while retaining a large majority of subjects (83 % of participants). Overall, results show it is possible to obtain usable resting-state data despite extreme motion in a group of young, untrained subjects., Competing Interests: Declaration of Competing Interest Damien A. Fair is a patent holder on the Framewise Integrated Real-Time Motion Monitoring (FIRMM) software. He is also a co-founder of Nous Imaging Inc. The nature of this financial interest have been reviewed by two committees at the University of Minnesota. They have put in place a plan to help ensure that his research is not affected by the financial interest., (Copyright © 2022 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Extended interval dosing of tobramycin is recommended for treatment of pulmonary exacerbations in adults and older children with cystic fibrosis (CF), but data are limited in patients less than 5 years of age. We performed a retrospective population pharmacokinetic (PK) analysis of hospitalized children with CF <5 years of age prescribed intravenous tobramycin for a pulmonary exacerbation from March 2011 to September 2018 at our hospital. Children with normal renal function who had ≥1 tobramycin concentration available were included. Nonlinear mixed effects population PK modeling was performed using NONMEM using data from the first 48 h of tobramycin treatment. Monte Carlo simulations were implemented to determine the fraction of simulated patients that met published therapeutic targets with regimens of 10-15 mg/kg/day once-daily dosing. Fifty-eight patients received 111 tobramycin courses (range 1-9/patient). A two-compartment model best described the data. Age, glomerular filtration rate, and vancomycin coadministration were significant covariates on tobramycin clearance. The typical values of clearance and central volume of distribution were 0.252 L/hr/kg^0.75 and 0.308 L/kg, respectively. No once-daily regimens achieved all pre-specified targets simultaneously in >75% of simulated subjects. A dosage of 13 mg/kg/dose best met the predefined targets of C max >25 mg/L and AUC 24 of 80-120 mg·h/L. Based on our population PK analysis and simulations, once-daily dosing of tobramycin would not achieve all therapeutic goals in young patients with CF. However, extended-interval dosing regimens may attain therapeutic targets in the majority of young patients.

Children's ability to discriminate nonsymbolic number (e.g., the number of items in a set) is a commonly studied predictor of later math skills. Number discrimination improves throughout development, but what drives this improvement is unclear. Competing theories suggest that it may be due to a sharpening numerical representation or an improved ability to pay attention to number and filter out non-numerical information. We investigate this issue by studying change in children's performance (N = 65) on a nonsymbolic number comparison task, where children decide which of two dot arrays has more dots, from the middle to the end of 1st grade (mean age at time 1 = 6.85 years old). In this task, visual properties of the dot arrays such as surface area are either congruent (the more numerous array has more surface area) or incongruent. Children rely more on executive functions during incongruent trials, so improvements in each congruency condition provide information about the underlying cognitive mechanisms. We found that accuracy rates increased similarly for both conditions, indicating a sharpening sense of numerical magnitude, not simply improved attention to the numerical task dimension. Symbolic number skills predicted change in congruent trials, but executive function did not predict change in either condition. No factor predicted change in math achievement. Together, these findings suggest that nonsymbolic number processing undergoes development related to existing symbolic number skills, development that appears not to be driving math gains during this period. Children's ability to discriminate nonsymbolic number improves throughout development. Competing theories suggest improvement due to sharpening magnitude representations or changes in attention and inhibition. The current study investigates change in nonsymbolic number comparison performance during first grade and whether symbolic number skills, math skills, or executive function predict change. Children's performance increased across visual control conditions (i.e., congruent or incongruent with number) suggesting an overall sharpening of number processing. Symbolic number skills predicted change in nonsymbolic number comparison performance., (© 2021 John Wiley & Sons Ltd.)

Introduction: Cystic fibrosis (CF)-related liver disease (CFLD) manifests as a wide spectrum of hepatobiliary disease and can progress to need liver transplantation. Elexacaftor/tezacaftor/ivacaftor (elx/tez/iva) is a cystic fibrosis transmembrane conductance regulator modulator that has superior efficacy compared to previously approved modulators. Use of elx/tez/iva, should be approached with caution in individuals with CFLD or following liver transplantation due to possible increases in liver function tests (LFTs) and drug-drug interactions with several immunosuppressant medications., Objective: The purpose of this case series is to explore if the use of elx/tez/iva is safe and tolerable in patients with CF postliver transplantation., Methods: A retrospective case series including patients prescribed elx/tez/iva following liver transplantation and an immunosuppressive regimen consisting of drug therapy metabolized by P-glycoprotein was completed., Results: Ten patients at six CF centers with a median age of 22.1 years (range 14-43.4 years) and the median time from the transplant of 6.9 years (range 0.6-22 years) were included. Most patients (8, 80%) received a reduced or full dose of elx/tez/iva for a mean duration of 10.4 months (range 7-12 months). Fluctuations in LFTs occurred in all patients (10, 100%) and led to therapy discontinuation in two patients (20%). Elx/tez/iva initiation resulted in elevations in tacrolimus trough concentration in seven patients (70%). Most patients who tolerated elx/tez/iva had symptomatic and quality of life improvement, increased body mass index, and maintained or improved lung function., Conclusion: Initiation of elx/tez/iva in patients with CF who received liver transplantation may be safe with clinical benefits., (© 2021 Wiley Periodicals LLC.)

The provision of high-quality early mathematics instruction and intervention is critical to ensure that all students are on track for academic success. Given this, identifying and utilizing assessments that can enable the detection of nonresponse to mathematics instruction is a critical aspect of early intervention. To this end, the current study explored the extent to which there were distinct patterns of performance on embedded assessments for intervention participants within the context of a large-scale randomized control trial of the ROOTS intervention. This study also examined how performance on embedded assessments was associated with pretest mathematics scores and residual gains on mathematics measures, and how those associations differed based on (a) the point in the intervention when students demonstrated difficulty, and (b) intervention intensity. Findings from this study suggest that participants fell into 4 distinct performance categories and performance classifications were associated with pretest measures and gains in mathematics achievement. Study results also highlight the potential relevance of instructional intensity and timely monitoring of student performance. Implications for intervention and instructional planning in the context of tiered instructional delivery models are discussed. (PsycINFO Database Record (c) 2019 APA, all rights reserved).

Objective: To identify whether slow aquatic exercise in the form of modified Ai Chi is more effective than conventional (faster pace) aquatic therapy at reducing arm volume in women with or at risk of breast cancer related lymphoedema., Methods: Randomized, cross-over controlled trial with concealed allocation and blinded assessment. Eighteen women with a history of breast cancer related lymphoedema were recruited. Participants received two intervention sessions (randomized order) with one week apart. Interventions were a 50min conventional aquatic intervention or a 50min modified Ai Chi. Arm volume was measured as the difference between affected and unaffected arm; bio-impedance was measured as an index of extracellular fluid; satisfaction was measured via a 12 question form. Outcomes were measured before, immediately after and one hour after intervention., Results: Comparison between interventions showed larger decreased arm volume of 140mL (95%CI 17-263) immediately after intervention in favor of the Ai Chi intervention, however it was not sustained at 1h follow-up. A post hoc analysis showed 72% of participants had a decrease in arm volume immediately after Ai Chi compared to 28% immediately after conventional aquatic therapy; with a number needed to treat of 3 (95%CI 1.4-6.6). There were no differences between interventions for bio-impedance. Satisfaction was good for both interventions., Conclusion: Slow pace aquatic exercise is more effective than conventional aquatic exercise immediately after intervention for arm volume. Also, undesirable increase in arm volume seems to subside after 1h, which can be beneficial if therapy does not address arm volume., Trial Registration: ACTRN12614000557639 (https://www.anzctr.org.au/Trial/Registration/TrialReview.aspx?ACTRN=12614000557639)., (Copyright © 2018 Associação Brasileira de Pesquisa e Pós-Graduação em Fisioterapia. Publicado por Elsevier Editora Ltda. All rights reserved.)

The purpose of this study was to investigate the incremental validity of a set of domain general cognitive measures added to a traditional screening battery of early numeracy measures. The sample consisted of 458 kindergarten students of whom 285 were designated as severely at-risk for mathematics difficulty. Hierarchical multiple regression results indicated that Wechsler Abbreviated Scales of Intelligence (WASI) Matrix Reasoning and Vocabulary subtests, and Digit Span Forward and Backward measures explained a small, but unique portion of the variance in kindergarten students' mathematics performance on the Test of Early Mathematics Ability-Third Edition (TEMA-3) when controlling for Early Numeracy Curriculum Based Measurement (EN-CBM) screening measures (R² change = .01). Furthermore, the incremental validity of the domain general cognitive measures was relatively stronger for the severely at-risk sample. We discuss results from the study in light of instructional decision-making and note the findings do not justify adding domain general cognitive assessments to mathematics screening batteries. (PsycINFO Database Record, ((c) 2018 APA, all rights reserved).)

Expression or introduction of the neuropeptide substance-P (SP; encoded by the TAC1 gene in humans and Tac1 in rodents) in the amygdala induces anxiety related behaviour in rodents. In addition, pharmacological antagonism of the main receptor of SP in humans; NK1, is anxiolytic. In the current study, we show that the Tac1 locus is up-regulated in primary rat amygdala neurones in response to activation of the glucocorticoid receptor (GR); a classic component of the stress response. Using a combination of bioinformatics, electrophoretic mobility shift assays (EMSA) and reporter plasmid magnetofection into rat primary amygdala neurones we identified a highly conserved GR response sequence (2GR) in the human TAC1 promoter that binds GR in response to dexamethasone (Dex) or forskolin. We also identified a second GR binding site in the human promoter that was polymorphic and whose T-allele is only found in Japanese and Chinese populations. We present evidence that the T-allele of SNPGR increases the activity of the TAC1 promoter through de-sequestration or de-repression of 2GR. The identification of Dex/forskolin response elements in the TAC1 promoter in amygdala neurones suggests a possible link in the chain of molecular events connecting GR activation and anxiety. In addition, the discovery of a SNP which can alter this response may have implications for our understanding of the role of regulatory variation in susceptibility to stress in specific populations., (Copyright © 2014 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Polymorphisms within intron 2 of the CNR1 gene, which encodes cannabinoid receptor 1 (CB(1)), have been associated with addiction, obesity, and brain volume deficits. We used comparative genomics to identify a polymorphic (rs9444584-C/T) sequence (ECR1) in intron 2 of the CNR1 gene that had been conserved for 310 million years. The C-allele of ECR1 (ECR1(C)) acted as an enhancer in hypothalamic and dorsal root ganglia cells and responded to MAPK activation through the MEKK pathway but not in hippocampal cells. However, ECR1(T) was significantly more active in hypothalamic and dorsal root ganglia cells but, significantly, and in contrast to ECR1(C), was highly active in hippocampal cells where it also responded strongly to activation of MAPK. Intriguingly, rs9444584 is in strong linkage disequilibrium with two other SNPs (rs9450898 (r(2) = 0.841) and rs2023239 (r(2) = 0.920)) that have been associated with addiction, obesity (rs2023239), and reduced fronto-temporal white matter volumes in schizophrenia patients as a result of cannabis misuse (rs9450898). Considering their high linkage disequilibrium and the increased response of ECR1(T) to MAPK signaling when compared with ECR1(C), it is possible that the functional effects of the different alleles of rs9444584 may play a role in the conditions associated with rs9450898 and rs2023239. Further analysis of the different alleles of ECR1 may lead to a greater understanding of the role of CNR1 gene misregulation in these conditions as well as chronic inflammatory pain.

The expression of the galanin gene (GAL) in the paraventricular nucleus (PVN) and in the amygdala of higher vertebrates suggests the requirement for highly conserved, but unidentified, regulatory sequences that are critical to allow the galanin gene to control alcohol and fat intake and modulate mood. We used comparative genomics to identify a highly conserved sequence that lay 42 kb 5' of the human GAL transcriptional start site that we called GAL5.1. GAL5.1 activated promoter activity in neurones of the PVN, arcuate nucleus and amygdala that also expressed the galanin peptide. Analysis in neuroblastoma cells demonstrated that GAL5.1 acted as an enhancer of promoter activity after PKC activation. GAL5.1 contained two polymorphisms; rs2513280(C/G) and rs2513281(A/G), that occurred in two allelic combinations (GG or CA) where the dominant GG alelle occurred in 70-83 % of the human population. Intriguingly, both SNPs were found to be in LD (R(2) of 0.687) with another SNP (rs2156464) previously associated with major depressive disorder (MDD). Recreation of these alleles in reporter constructs and subsequent magnetofection into primary rat hypothalamic neurones showed that the CA allele was 40 % less active than the GG allele. This is consistent with the hypothesis that the weaker allele may affect food and alcohol preference. The linkage of the SNPs analysed in this study with a SNP previously associated with MDD together with the functioning of GAL5.1 as a PVN and amygdala specific enhancer represent a significant advance in our ability to understand alcoholism, obesity and major depressive disorder.

The neuropeptide substance-P (SP) is expressed from the TAC1 gene in sensory neurones where it acts as a key modulator of neurogenic inflammation. The promoter of TAC1 (TAC1prom) plays a central role in the regulation of the TAC1 gene but requires the presence of a second regulatory element; ECR2, to support TAC1 expression in sensory neurones and to respond appropriately to signalling pathways such as MAPkinases and noxious induction by capsaicin. We examined whether the effect of capsaicin on ECR2-TAC1prom activity in larger diameter neurones was cell autonomous or non- cell autonomous. We demonstrate that TRPV1 is not expressed in all the same cells as SP following capsaicin induction suggesting the presence of a non-cell autonomous mechanism for TAC1 up-regulation following capsaicin induction. In addition, we demonstrate that induction of SP and ECR1-TAC1prom activity in these larger diameter neurones can be induced by potassium depolarisation suggesting that, in addition to capsaicin induction, transgene activity may be modulated by voltage gated calcium channels. Furthermore, we show that NK1 is expressed in all SP- expressing cells after capsaicin induction and that an agonist of NK1 can activate both SP and the transgene in larger diameter neurones. These observations suggest the presence of an autocrine loop that controls the expression of the TAC1 promoter in sensory neurones. In contrast, induction of the TAC1 promoter by LPS was not dependent on ECR2 and did not occur in large diameter neurones. These studies demonstrate the diversity of mechanisms modulating the activity of the TAC1 promoter and provide novel directions for the development of new anti-inflammatory therapies.

Changes in the expression of the neuropeptide substance P (SP) in different populations of sensory neurones are associated with the progression of chronic inflammatory disease. Thus, understanding the genomic and cellular mechanisms driving the expression of the TAC1 gene, which encodes SP, in sensory neurones is essential to understanding its role in inflammatory disease. We used a novel combination of computational genomics, primary-cell culture and mouse transgenics to determine the genomic and cellular mechanisms that control the expression of TAC1 in sensory neurones. Intriguingly, we demonstrated that the promoter of the TAC1 gene must act in synergy with a remote enhancer, identified using comparative genomics, to respond to MAPK signalling that modulates the expression of TAC1 in sensory neurones. We also reveal that noxious stimulation of sensory neurones triggers this synergy in larger diameter sensory neurones--an expression of SP associated with hyperalgesia. This noxious stimulation of TAC1 enhancer-promotor synergy could be strongly blocked by antagonism of the MEK pathway. This study provides a unique insight into the role of long-range enhancer-promoter synergy and selectivity in the tissue-specific response of promoters to specific signal transduction pathways and suggests a possible new avenue for the development of novel anti-inflammatory therapies., (Copyright © 2010 S. Karger AG, Basel.)

Effectively directed neuron migration is critical for development and repair in the central nervous system (CNS). Endogenous electric fields (EFs) are widespread in developing and regenerating tissues and regulate a variety of cell behaviors including directed cell migration. Electrically-directed neuronal migration has not been tested previously and we show that an applied EF directs migration of hippocampal neurons toward the cathode at a field strength of 120 mV/mm, close to the physiological range. Reversal of the field polarity reversed the direction of neuron migration. Neuron migration from an explant also was directed by an applied EF. Mechanistically, EF-guided migration was transduced by activation of the second messenger molecules ROCK (Rho-associated protein kinase) and PI3 kinase (phosphoinositide-3 kinase) since their pharmacological inhibition decreased the directedness and speed of neuron migration. This work demonstrates that rat hippocampal neurons respond to applied EFs with directional migration and raises the possibility that EFs may be used as a cue to direct neuronal migration in novel strategies to repair the CNS., ((c) 2008 Wiley-Liss, Inc.)

It is well documented that the hormone leptin signals information regarding the status of fat stores to hypothalamic nuclei, which in turn control feeding behaviour and body weight. However, leptin and its receptor are widely expressed in many extra-hypothalamic brain regions, including hippocampus, brain stem and cerebellum. Moreover, evidence is accumulating that leptin has other neuronal functions that are unrelated to its effects on energy homeostasis. Indeed a role for leptin in neuronal development has been suggested as leptin-deficient rodents display abnormal brain development and leptin actively participates in the development of the hypothalamus. In the hippocampus, leptin is a potential cognitive enhancer as genetically obese rodents with dysfunctional leptin receptors display impairments in hippocampal synaptic plasticity. Moreover, direct administration of leptin into the hippocampus can facilitate hippocampal LTP (long-term potentiation) in vivo and improve memory processing in mice. At the cellular level, we have also shown that leptin has the capacity to convert short-term potentiation into LTP. Here, we review the data that leptin influences hippocampal synaptic plasticity via enhancing NMDA (N-methyl-D-aspartate) receptor function. We also provide evidence that rapid trafficking of NMDA receptors to the plasma membrane may underlie the effects of leptin on excitatory synaptic strength.

Motor nerve terminals in adult mammalian slow-twitch muscles have lower levels of spontaneous and evoked neurotransmitter release than terminals in fast-twitch muscles. These reflect adaptive differences, allowing terminals in slow (postural) muscles to sustain release during the prolonged firing trains experienced in vivo. Here we ask whether these differences in terminal release properties in Sprague-Dawley rat extensor digitorum longus (EDL, fast) and soleus (slow) muscles reflect their early cytodifferentiation in the embryo or whether they might be adaptations to their distinct mature activity patterns, which emerge around two weeks postnatally. We find that the mature pattern of differences in release arise through co-ordinated increases in presynaptically dependent release properties (quantal content, spontaneous release frequency and evoked potential amplitude), beginning at three weeks, which are particularly substantial in EDL. In contrast, other synaptic properties are either consistently greater in the same muscle throughout development (evoked potential kinetics, muscle fibre diameter) or display no systematic muscle type-dependent differences (terminal area, input resistance, spontaneous release amplitude). Thus, the emergence of adaptive differences in terminal release properties correlates with the differentiation of locomotor activity patterns in postnatal rat hindlimb muscles.

The obese gene product leptin is an important signaling protein that regulates food intake and body weight via activation of the hypothalamic leptin receptor (Ob-Rb; Jacob et al., 1997). However, there is growing evidence that Ob-Rb is also expressed in CNS regions, not directly associated with energy homeostasis (Mercer et al., 1996; Hakansson et al., 1998). In the hippocampus, an area of the brain involved in learning and memory, we have found that leptin facilitates the induction of synaptic plasticity. Leptin converts short-term potentiation of synaptic transmission induced by primed burst stimulation of the Schaffer collateral commissural pathway into long-term potentiation. The mechanism underlying this effect involves facilitation of NMDA receptor function because leptin rapidly enhances NMDA-induced increases in intracellular Ca(2+) levels ([Ca(2+)](i)) and facilitates NMDA, but not AMPA, receptor-mediated synaptic transmission. The signaling mechanism underlying these effects involves activation of phosphoinositide 3-kinase, mitogen-activated protein kinase, and Src tyrosine kinases. These data indicate that a novel action of leptin in the CNS is to facilitate hippocampal synaptic plasticity via enhanced NMDA receptor-mediated Ca(2+) influx. Impairment of this process may contribute to the cognitive deficits associated with diabetes mellitus.

Despite large environmental variations, the human body maintains a tightly regulated core temperature. Effective thermoregulation must balance the interaction between skin surface, clothing and ambient air. Indices of thermal stress (wet bulb globe temperature, heat stress index, maximum evaporation rate, required evaporative rate and wind chill) provide valuable information concerning the heat exchange between the individual and the environment, and serve as protective guidelines while working in environmental extremes. The role of clothing, as an interactive barrier, greatly affects thermal balance. Clothing is varied according to prevailing environmental conditions, metabolic heat production, gender and age differences, fabric thermal properties, garment design and intended use. Models (static, dynamic and human) have investigated the biophysical transfer of heat between the skin surface area, clothing and ambient air. Additionally, the role of metabolic heat production during exercise can greatly influence tolerance to thermal stress during a variety of environmental conditions.

We studied the scores obtained on the Millon Clinical Multiaxial Inventory (MCMI) by Black and White male psychiatric inpatients to determine the presence or absence of racial bias. In predicting psychopathology for the two races, comparisons of MCMI performance indicated significant differences for all diagnoses except the personality disorders. The subjects were then matched into two groups of 209 patients each, according to DSM-III psychiatric diagnoses. The data were analyzed at the item, scale, and structural levels. At the item level, application of the Mantel-Haenszel Procedure revealed that 45 of the 175 items of the inventory were answered significantly different by the two racial groups. Because this number was higher than what could be expected by chance, the finding suggested possible deficiencies in terms of the culture-fairness of the items used in the test. At the scale level, an analysis of variance (ANOVA) demonstrated that the scores obtained by the Black and White groups were significantly different in 9 of the 20 scales (Histrionic, Narcissistic, Antisocial, Paraphrenia, Hypomania, Dysthymia, Alcohol Abuse, Drug Abuse, and Psychotic Delusion). With the exception of the Dysthymic scale, all of the differences were in the direction of the Blacks obtaining a higher score than the Whites. At the structural level, however, a principal components factor analysis performed on each group resulted in factor structures that looked identical.