Similar Functional Changes Research Articles

Maternal Diabetes and Cardiac Left Ventricular Structure and Function in the Infant: A Copenhagen Baby Heart Study.

Prenatal exposure to maternal diabetes is associated with an increased risk of offspring heart defects. We evaluated associations with subtle infant cardiac changes. In a cohort of 25,486 infants with transthoracic echocardiography within 60 days of birth, we investigated associations between maternal preexisting diabetes and gestational diabetes mellitus (GDM) and infant left ventricular (LV) structural and functional parameters, using linear regression to estimate adjusted mean differences (aMDs) between groups. Infants exposed to maternal preexisting diabetes (n = 198) had thicker LV posterior walls (aMD 0.19 mm; 95% CI 0.11, 0.27), smaller LV internal diameters in systole (aMD -0.27 mm; 95% CI -0.45, -0.18) and diastole (aMD -0.37 mm; 95% CI -0.59, -0.09), reduced stroke volumes (aMD -0.36 mL; 95% CI -0.61, -0.11), and increased heart rates (aMD 3.14 bpm; 95% CI 1.10, 6.18) and mitral valve early peak velocities (aMD 2.17 cm/s; 95% CI 0.31, 4.04) than unexposed infants (n = 24,639). Infants born to mothers with GDM (n = 649) had significantly smaller LV internal diameters in systole (aMD -0.13 mm; 95% CI -0.22, -0.03) and similar structural and functional changes as children exposed to preexisting diabetes, albeit with smaller nonsignificant aMDs. Higher third-trimester HbA1c levels were associated with smaller LV internal diameters and stroke volumes in infants exposed to preexisting diabetes and with lower heart rates in infants exposed to GDM. Maternal preexisting diabetes and, to a lesser extent, GDM were associated with changes in infant LV structure and function.

Microglial priming by IFN-γ involves STAT1-mediated activation of the NLRP3 inflammasome.

Inflammatory and immune responses in the brain that contribute to various neuropsychiatric disorders may begin as microglial "priming". Interferon (IFN)-γ is known to cause microglial priming, but the mechanism is unclear. We examined the effects of IFN-γ on gene expression, microglial activation, inflammatory and immune responses and activity of the NLRP3 inflammasome in primary microglia and in the brains of mice. Our results showed that treating microglial cultures with IFN-γ induced a hedgehog-like morphology and upregulated markers of microglial activation (CD86, CD11b) and pro-inflammatory molecules (IL-1β, IL-6, TNF-α, iNOS), while downregulating markers of microglial homeostasis (CX3CR1, CD200R1), anti-inflammatory molecules (MCR1, Arg-1) and neurotrophic factors (IGF-1, BDNF). IFN-γ also upregulated markers of NLRP3 inflammasome activation (NLRP3, caspase-1, gasdermin D, IL-18). This particular transcriptional profiling makes IFN-γ-primed microglia with exaggerated responses upon lipopolysaccharide (LPS) stimulation. The level of NLRP3, caspase-1, gasdermin D, IL-1β, IL-18, TNF-α and iNOS in microglia cultures treated with both IFN-γ and LPS were highest than with either one alone. Injecting IFN-γ into the lateral ventricle of mice induced similar morphological and functional changes in hippocampal microglia as in primary microglial cultures. The effects of IFN-γ on NLRP3 inflammasome and microglia from cultures or hippocampus were abolished when STAT1 was inhibited using fludarabin. Injecting mice with IFN-γ alone or together with LPS induced anxiety- and depression-like behaviors and impaired hippocampus-dependent spatial memory; these effects were mitigated by fludarabin. IFN-γ primes microglia by activating STAT1, which upregulates genes that activate the NLRP3 inflammasome. Inhibiting the IFN-γ/STAT1 axis may be a way to treat neurodegenerative diseases and psychiatric disorders that involve microglial priming.

Tuning the Response of Synthetic Mechanogenetic Gene Circuits Using Mutations in TRPV4.

Conventional gene therapy approaches for drug delivery generally rely on constitutive expression of the transgene and thus lack precise control over the timing and magnitude of delivery. Synthetic gene circuits with promoters that are responsive to user-defined stimuli can provide a molecular switch that can be utilized by cells to control drug production. Our laboratory has previously developed a mechanogenetic gene circuit that can deliver biological drugs, such as interleukin-1 receptor antagonist (IL-1Ra), on-demand through the activation of Transient receptor potential family, vanilloid 4 (TRPV4), a mechanosensory ion channel that has been shown to be activated transiently in response to physical stimuli such as physiological mechanical loading or hypo-osmotic stimuli. The goal of this study was to use mutations in TRPV4 to further tune the response of this mechanogenetic gene circuit. Human iPSC-derived chondrocytes harboring targeted gain-of-function mutations of TRPV4 were chondrogenically differentiated. Both mutants-V620I and T89I-showed greater total IL-1Ra production compared with wild type following TRPV4 agonist treatment, as well as mechanical or osmotic loading, but with altered temporal dynamics. Gene circuit output was dependent on the degree of TRPV4 activation secondary to GSK101 concentration or strain magnitude during loading. V620I constructs secreted more IL-1Ra compared with T89I across all experimental conditions, indicating that two mutations that cause similar functional changes to TRPV4 can result in distinct circuit activation profiles that differ from wild-type cells. In summary, we successfully demonstrate proof-of-concept that point mutations in TRPV4 that alter channel function can be used to tune the therapeutic output of mechanogenetic gene circuits.

High copper levels induce premature senescence in 3T3-L1 preadipocytes

Copper (Cu) dyshomeostasis has been linked to obesity and related morbidities and also to aging. Cu levels are higher in older or obese individuals, and adipose tissue (AT) Cu levels correlate with body mass index. Aging and obesity induce similar AT functional and structural changes, including an accumulation of senescent cells. To study the effect of Cu-mediated stress-induced premature senescent (Cu-SIPS) on preadipocytes, 3T3-L1 cell line was exposed to a subcytotoxic concentration of copper sulfate. After Cu treatment, preadipocytes acquired typical senescence characteristics including diminished cell proliferation, cell and nuclei enlargement and increased lysosomal mass (higher Lamp2 expression and a slight increased number of cells positive for β-galactosidase associated with senescence (SA-β-Gal)). Cell cycle arrest was due to upregulation of p16Ink4aInk4a and p21Waf1/Cip1. Accordingly, protein levels of the proliferation marker KI67 were reduced. Cu-SIPS relates with oxidative stress and, in this context, an increase of SOD1 and HO-1 expression was detected in Cu-treated cells. The mRNA expression of senescence-associated secretory phenotype factors, such as Mmp3, Il-6 and Tnf-α, increased in Cu-SIPS 3T3-L1 cells but no effect was observed on the expression of heterochromatin-associated protein 1(HP1).Although the downregulation of Lamin B1 expression is considered a hallmark of senescence, Cu-SIPS cells presented higher levels of Lamin B1. The dysregulation of nuclear lamina was accompanied by an increase of nuclear blebbing, but not of micronuclei number.To conclude, a Cu-SIPS model in 3T3-L1 preadipocytes is here described, which may be an asset to the study of AT dysregulation observed in obesity and aging.

Pharmacological and Psychological Treatment Have Common and Specific Effects on Brain Activity in Obsessive-Compulsive Disorder

Initial treatment for obsessive-compulsive disorder (OCD) consists of pharmacological treatment with selective serotonin reuptake inhibitors (SSRIs) and/or psychological treatment with cognitive behavioral therapy (CBT). The assumption is that both treatments have different neural working mechanisms, but empirical evidence is lacking. We investigated whether these treatments induce similar or different functional neural changes in OCD. We conducted a longitudinal nonrandomized controlled trial in which thirty-four OCD patients were treated with sixteen weeks of CBT or SSRIs. Functional magnetic resonance imaging was performed before and after treatment during emotional processing (emotional face matching and symptom provocation tasks) and response inhibition (stop signal task). Twenty matched healthy controls were scanned twice with a similar time interval. Both CBT and SSRIs were successful in reducing OCD symptoms. Compared to healthy controls, treatment led to a reduction of insula activity in OCD patients during symptom provocation. The comparison between treatment groups revealed widespread divergent brain changes in the cerebellum, posterior insula, caudate nucleus, hippocampus, and occipital and prefrontal cortex during all tasks, explained by relative increases of activity following CBT compared to relative decreases of activity following SSRIs. Pharmacological and psychological treatment primarily lead to opposite changes in brain function, with a common reduction of insula activity during symptom provocation. These findings provide insight into common and specific neural mechanisms underlying treatment response, suggesting that CBT and SSRIs support recovery from OCD along partly distinct pathways. This trial is registered with NTR6575.

Association between migraine and the risk of vascular dementia: A nationwide longitudinal study in South Korea.

We aimed to examine the potential association between migraine and vascular dementia (VaD) using a nationwide population database. Migraine and VaD showed similar structural and functional changes in pathophysiology process and shared common risk factors, However, whether migraine prevalence increases VaD incidence remains controversial. This retrospective population-based cohort study used the medical records from the Korean National Health Insurance System database. Migraine (G43) was defined by using the Tenth Revision of the International Classification of Diseases code. More than two migraine diagnoses at least 3 months apart were defined as "chronic migraine". Cox proportional hazards model estimated hazard ratios (HRs) of VaD for group comparisons. We included 212,836 patients with migraine and 5,863,348 individuals without migraine. During 10 years of follow-up, 3,914 (1.8%) and 60,258 (1.0%) patients with and without migraine, respectively, were newly diagnosed with VaD. After adjustment, patients with migraine showed a 1.21-fold higher risk of VaD than those without migraine (HR = 1.21; 95% confidence interval (CI): 1.17-1.25). Patients with chronic migraine showed a higher cumulative incidence of VaD than those with episodic migraine. The adjusted HR for the VaD incidence with migraine was higher in: (1) patients aged <65 years; (2) women; (3) patients without hypertension, diabetes, or atrial fibrillation; and (4) non-smokers. Migraine is associated with an increased risk of VaD, particularly in chronic migraine patients. Incidence of VaD in the setting of migraine may have distinct pathophysiology from that of VaD with traditional cardiovascular risks.

Abstract P1107: Elmsan1 Regulates Differentiation And Maturation Of Cardiomyocytes Derived From Human Induced Pluripotent Stem Cells

Introduction: ELMSAN1 is a scaffolding protein of the mitotic deacetylase complex (MiDAC) that is critical for early embryonic development. Studies on Elmsan1 knockout mice show that its absence results in heart malformation and embryo lethality. Despite this, the direct role of ELMSAN1 in heart development and formation remains unknown. To gain insights into this role, we differentiated human-induced pluripotent stem cell (hiPSC) into cardiomyocyte (hiPSC-CM) to mimic early cardiogenesis and assess the involvement of ELMSAN1 in CM maturation. Methods and Results: We utilized shRNA-mediated knockdown and CRISPR-Cas9-mediated knockout techniques to generate ELMSAN1-deficient hiPSC, which were differentiated into CM for characterization. We showed that ELMSAN1 depletion caused a delay in the deactivation of pluripotency genes during early differentiation, resulting in decreased expression of cardiac-specific markers (TNNT2, MHY6), and reduced differentiation efficiency of CM. We also observed the impaired expression of genes associated with contractile sarcomere structure, calcium handling, and ion channels in ELMSAN1-deficient CM. Moreover, through a series of structural and functional assessments, we found these cells exhibited incomplete sarcomere Z-line structure and length (1.3 μm in KD vs. 1.5 μm in control), decreased calcium amplitude, increased time to peak value, faster beat rate with a shorter beat period, decreased field potential duration (300 ± 10 ms in KD vs. 500 ± 10 ms in control), and prolonged propagation delay. Importantly, we confirmed similar immaturity and functional changes in ELMSAN1 knockout CM, excluding the potential off-target effect. Mechanistically, we discovered that ELMSAN1 depletion resulted in decreased levels of H3K27ac in both hiPSC and CM, indicating an unexpected role of ELMSAN1 in maintaining global H3K27ac and gene expression. Conclusion: Our findings offer compelling evidence for the direct involvement of ELMSAN1 in the differentiation and maturation of hiPSC-CM. Notably, we were the first to identify the impact of ELMSAN1 on various aspects of hiPSC-CM generation, including cardiac differentiation, sarcomere formation, calcium handling, and electrophysiological maturation.

ПРОИСХОЖДЕНИЕ РЕЛИГИОЗНЫХ ПРЕДСТАВЛЕНИЙ ИЗ ОКОЛОСМЕРТНОГО ОПЫТА

The subject of the article is the idea that religious beliefs are originated from near-death experience. Firstly, it is supported by the existence of a necessary connection between mystical (religious) and near-death experience: 1) the majority of people who have survived clinical death, in their near-death experience have had an experience traditionally considered religious; 2) a clear correla-tion of brain activation in delta-, alpha- and gamma-bands has been established for both groups of near-death and religious experi-ences; 3) near-death and mystical experiences are associated with similar functional changes in the autonomic nervous system. Secondly, it’s corroborated by the correspondence between the complex of near-death experience and religious ideas about the "afterlife". The opposite modalities of subjective reality in the state of clinical death are oniric experiences similar to religious images of Hell and Paradise. Thirdly, it is supported by the origin of the majority of archaic religious ideas from near-death experience: 1) there are numerous direct evidences that religious ideas come from near-death experience; 2) the variability of the genesis of reli-gious ideas from analogs of near-death experience has been recorded; 3) the relationship between shamanic practices and near-death experience has been established.

The effects of drought treatments on biosynthesis and structure of maize starches with different amylose content

We investigated the effects of drought stress (DS) on maize varieties with different amylose content (AC). In starches with AC of 33 %, DS increased the contents of amylopectin (AP) chains with a degree of polymerization (DP) > 36 and decreased the AP chains with DP ≤ 36, while the AC was unchanged. DS decreased the crystallinity, the thickness of both amorphous and crystalline lamellae, and average granular size. In contrast, the digestibility increased. For starches with AC of 45 %, DS increased the content of AP chains with DP > 24 and AC, while the contents of AP chains with DP ≤ 24 decreased. DS produced starch with thinner crystalline lamellae, thicker amorphous lamellae, more elongated and larger granules. The digestibility of the starches decreased. In starches with AC of 53 %, moderate DS led to similar structural and functional changes as found for starches with AC of 45 %. Finally, severe DS resulted in the decrease of AC.

Reduced arterial elasticity after anabolic–androgenic steroid use in young adult males and mice

High-doses of anabolic–androgenic steroids (AAS) is efficient for building muscle mass, but pose a risk of cardiovascular side effects. Little is known of the effect of AAS on vasculature, but previous findings suggest unfavorable alterations in vessel walls and vasoreactivity. Here, long-term effect of AAS on vascular function and morphology were examined in male weightlifters, and in a mimicking animal model. Arterial elasticity and morphology were tested with ultrasound, pulse wave velocity (PWV) and carotid intima media thickness (cIMT) in 56 current male AAS users, and 67 non-exposed weightlifting controls (WLC). Female mice were treated with testosterone for 14 days and echocardiography were applied to evaluate vascular function and morphology. Male AAS users had higher PWV (p = 0.044), reduced carotid artery compliance (p = 0.0005), and increased cIMT (p = 0.041) compared to WLC. Similar functional changes were found in the ascending aorta of mice after 7- (p = 0.043) and 14 days (p = 0.001) of testosterone treatment. This animal model can be used to map molecular mechanisms responsible for complications related to AAS misuse. Considering the age-independent stiffening of major arteries and the predictive power of an increase in PWV and cIMT, the long-term users of AAS are at increased risk of severe cardiovascular events.

Gain of function due to increased opening probability by two KCNQ5 pore variants causing developmental and epileptic encephalopathy

Developmental and epileptic encephalopathies (DEEs) are neurodevelopmental diseases characterized by refractory epilepsy, distinct electroencephalographic and neuroradiological features, and various degrees of developmental delay. Mutations in KCNQ2, KCNQ3, and, more rarely, KCNQ5 genes encoding voltage-gated potassium channel subunits variably contributing to excitability control of specific neuronal populations at distinct developmental stages have been associated to DEEs. In the present work, the clinical features of two DEE patients carrying de novo KCNQ5 variants affecting the same residue in the pore region of the Kv7.5 subunit (G347S/A) are described. The in vitro functional properties of channels incorporating these variants were investigated with electrophysiological and biochemical techniques to highlight pathophysiological disease mechanisms. Currents carried by Kv7.5 G347 S/A channels displayed: 1) large (>10 times) increases in maximal current density, 2) the occurrence of a voltage-independent component, 3) slower deactivation kinetics, and 4) hyperpolarization shift in activation. All these functional features are consistent with a gain-of-function (GoF) pathogenetic mechanism. Similar functional changes were also observed when the same variants were introduced at the corresponding position in Kv7.2 subunits. Nonstationary noise analysis revealed that GoF effects observed for both Kv7.2 and Kv7.5 variants were mainly attributable to an increase in single-channel open probability, without changes in membrane abundance or single-channel conductance. The mutation-induced increase in channel opening probability was insensitive to manipulation of membrane levels of the critical Kv7 channel regulator PIP2. These results reveal a pathophysiological mechanism for KCNQ5-related DEEs, which might be exploited to implement personalized treatments.

98 Plasma Extracellular Vesicles Released After Sever Burn Injury Modulate Macrophage Function

IntroductionExtracellular microvesicles (MVs) have emerged as key regulators of immune function across multiple diseases and potential biomarkers. Severe burn injury is a devastating trauma with significant immune dysfunction that results in an ~12% mortality rate due to sepsis-induced organ failure, pneumonia, and other infections. Severe burn causes a biphasic immune response: an early (0-72 hrs) hyper-inflammatory state, with release of pro-inflammatory damage-associated molecular pattern molecules (DAMPs), such as HMGB1, and cytokines (e.g.IL-1b), followed by an immunosuppressive state (1-2 weeks post injury), associated with increased susceptibility to life-threatening infections. We have reported that early after severe burn injury HMGB1 and IL-1b are enriched in plasma microvesicles (MVs), suggesting a role for MVs in post-burn immune activation. Here we tested the impact of MVs isolated after burn injury on phenotypic and functional consequences in vivo and in vitro using adoptive MV transfers.MethodsWe then assessed if the cargo of MVs following burn injury in humans could predict length of hospital stay. MVs isolated early from mice that underwent a 20% total body surface area (TBSA) burn injury (burn MVs) caused similar cytokine responses in naïve mice to those seen in burned mice early after injury. Burn MVs transferred to RAW264.7 macrophages caused similar functional (i.e. cytokine secretion) and genetic changes (measured by NanoStringTM) seen with their associated phase of post-burn immune dysfunction.ResultsBurn MVs isolated early (24h) induced MCP-1, IL-12p70 and IFNg, while MVs isolated later (1 and 2 weeks) blunted RAW proinflammatory responses to bacterial endotoxin (LPS). Unbiased LC-MS / MS proteomic analysis of early EVS (< 72 h post-injury) showed similarities in human and mice.ConclusionsIn our sample of large burn injury, EV SAA1 and CRP correlated with TBSA injury in both sexes and were correlated with length of hospital stay in women.

Effects of Resting vs. Continuous Blood-Flow Restriction-Training on Strength, Fatigue Resistance, Muscle Thickness, and Perceived Discomfort.

Introduction: The purpose of this study was to clarify whether blood-flow restriction during resting intervals [resting blood-flow restriction (rBFR)] is comparable to a continuous BFR (cBFR) training regarding its effects on maximum strength, hypertrophy, fatigue resistance, and perceived discomfort. Materials and Methods: Nineteen recreationally trained participants performed four sets (30-15-15-15 repetitions) with 20% 1RM on a 45° leg press twice a week for 6 weeks (cBFR, n = 10; rBFR, n = 9). Maximum strength, fatigue resistance, muscle thickness, and girth were assessed at three timepoints (pre, mid, and post). Subjective pain and perceived exertion were determined immediately after training at two timepoints (mid and post). Results: Maximum strength (p < 0.001), fatigue resistance (p < 0.001), muscle thickness (p < 0.001), and girth (p = 0.008) increased in both groups over time with no differences between groups (p > 0.05). During the intervention, the rBFR group exposed significantly lower perceived pain and exertion values compared to cBFR (p < 0.05). Discussion: Resting blood-flow restriction training led to similar gains in strength, fatigue resistance, and muscle hypertrophy as cBFR training while provoking less discomfort and perceived exertion in participants. In summary, rBFR training could provide a meaningful alternative to cBFR as this study showed similar functional and structural changes as well as less discomfort.

Atrial strain in cardiacATTR amyloidosis from pathophysiology to prognosis: is it time to rethink our approach to disease?

Abstract Background Left atrial (LA) involvement in cardiac ATTR amyloidosis (ATTR-CM) has never being explored. The aim of the study is to characterize the spectrum of changes in LA structure in ATTR-CM, the functional consequences on LA mechanics and the association with mortality. Methods We studied 936 patients with ATTR-CM who underwent prospective protocolized evaluations comprising ECG, full echocardiographic assessment, 6MWT, blood biomarkers sampling and survival assessment between 2000 and 2019. Reservoir (LAr), conduit (LAcd) and booster (LAcont) function were assessed according to current guidelines. LA stiffness (LAstiff) was estimated as the ratio between E/e' and LAr. The presence and extent of amyloid infiltration was assessed in two autopsied hearts from patients ATTR-CM with congo red staining and TTR antibodies. Results 936 patients with ATTR-CM were included, 567 with wtATTR-CM and 346 with hATTR-CM, of whom 246 had the V122I variant and 100 the T60A variant. LA indexed area was similar across the 3 genotypes, whilst LAr, LAcd and LAcont were significantly reduced in V122I patients compared to Wt and T60A patients (LAr: 10.4±6.4 vs 12.8±8.5 vs 8.59±5.7%, p&amp;lt;0.0001; LAcd: 5.6±5.0 vs 6.9±5.7 vs 4.0±3.6%, p&amp;lt;0.0001; LAcont 7.5±4.1 vs 8.2±4.9 vs 6.7±4.2%, p=0.023; in Wt, T60 and V122I respectively). Similarly, LAstiff was significantly worse in V122I compared to Wt and T60A (LAstiff: 2.39±2.62 vs 2.59±4.83 vs 2.96±2.88, p&amp;lt;0.0001, in Wt, T60 and V122I respectively). In the overall population, LAstiff remained independently associated with mortality, after adjusting for rhythm, systolic and diastolic functional parameters, ventricular longitudinal strain, genotypes, clinical variables and NAC staging system (lnLA stiff: HR=1.26, CI: 1.07–1.57; p=0.009). In the overall population, 584 patients were in sinus rhythm and 344 in atrial fibrillation, atrial flutter or atrial tachycardia (non-sinus rhythm group – NSR). Among patients in SR, 21.6% did not show a measurable LA mechanical contraction on strain analysis (LAcont). Patients in SR without LAcont on strain analysis showed similar structural and functional LA changes, NTproBNP, NYHA class, and 6MWT to NSR patients. Mortality of patients in SR with no LAcont was significantly reduced compared to patients in SR with LAcont and similar to patients in AF (Figure 1). On the two autopsies hearts there was extensive amyloid infiltration of TTR type in the atrial wall as confirmed with CR and TTR antibodies (Figure 1). Conclusions ATTR-CM is characterized by primary infiltration of the atrial walls with progressive loss of atrial function and increased stiffness. LA stiffness is a strong independent predictor of mortality after adjusting for known predictors. Atrial electro-mechanical dissociation emerged as a distinctive functional phenotype identifying patients in SR with worse prognosis. Figure 1 Funding Acknowledgement Type of funding source: None

Interaction of Manganese and Ammonia in the Brain of Hepatic Encephalopathy Rats

Background: Both ammonia and manganese (Mn) play a key role in the pathogenesis of hepatic encephalopathy (HE) and cause similar morphological and functional changes in astrocytes. Objectives: To investigate the interaction between brain Mn and ammonia in HE rats. Methods: Three rat models of minimal HE (MHE), chronic manganism (CHM), and chronic hyperammonemia (CHA) were constructed. A total of 48 Sprague-Dawley rats were divided into one control group (n = 6), MHE groups (n = 18, among which six rats were used to evaluate the MHE model), CHM groups (n = 12), and CHA groups (n = 12). The CHM, CHA, and the rest of MHE rats were randomly divided further into two subgroups, according to the MgSO4 treatment (oral administration of 496 mg/kg/day for seven weeks): MHE-7W and MHE + Mg-7W; CHM-7W and CHM + Mg-7W; and CHA-7W and CHA + Mg-7W, respectively. Rats’ blood ammonia, brain Mn, glutamine synthetase (GS), and glutamine (GLN) levels were measured and compared among groups. Results: Significantly higher brain Mn content in MHE-7W and CHM-7W rats, higher blood ammonia levels, brain GS activity, and GLN content were observed in MHE-7W, CHM-7W, and CHA-7W rats than in control rats. After MgSO4 treatment for seven weeks, significantly lower brain Mn content, blood ammonia levels, and GLN content were observed in MHE, CHM, and CHA rats. Conclusions: Our study showed that brain Mn accumulation could increase brain ammonia levels, while the accumulation of brain ammonia had no effect on the content of brain Mn.

267 – Exclusive Enteral Nutrition (EEN) and Crohn's Disease Exclusion Diet (CDED) Produce Broadly Similar Taxonomic and Functional Changes During the Induction of Remission, with Een Showing a Pattern of Rebound After Return to Free Diet+Partial Enteral Nutrition (PEN)

267 – Exclusive Enteral Nutrition (EEN) and Crohn's Disease Exclusion Diet (CDED) Produce Broadly Similar Taxonomic and Functional Changes During the Induction of Remission, with Een Showing a Pattern of Rebound After Return to Free Diet+Partial Enteral Nutrition (PEN)

Appendages and gene regulatory networks: Lessons from the limbless.

Among squamate reptiles, dozens of lineages have independently evolved complete or partial limb reduction. This remarkable convergence of limbless and limb-reduced phenotypes provides multiple natural replicates of different ages to explore the evolution and development of the vertebrate limb and the gene regulatory network that controls its formation. The most successful and best known of the limb-reduced squamates are snakes, which evolved a limb-reduced body form more than 100 million years ago. Recent studies have revealed the unexpected finding that many ancient limb enhancers are conserved in the genomes of snakes. Analyses in limbed animals show that many of these limb enhancers are also active during development of the phallus, suggesting that these enhancers may have been retained in snakes due their importance in regulating transcription in the external genitalia. This hypothesis is substantiated by functional tests of snake enhancers, which demonstrate that snake enhancer elements have lost limb function while retaining genital enhancer function. The large degree of overlap in the gene regulatory networks deployed during limb and phallus development may act to constrain the divergence of shared gene network components and the evolution of appendage morphology. Future studies will reveal whether limb regulatory elements have undergone similar functional changes in other lineages of limb-reduced squamates.

Human APP Gene Expression Alters Active Zone Distribution and Spontaneous Neurotransmitter Release at the Drosophila Larval Neuromuscular Junction.

This study provides further insight into the molecular mechanisms that control neurotransmitter release. Experiments were performed on larval neuromuscular junctions of transgenic Drosophila melanogaster lines with different levels of human amyloid precursor protein (APP) production. To express human genes in motor neurons of Drosophila, the UAS-GAL4 system was used. Human APP gene expression increased the number of synaptic boutons per neuromuscular junction. The total number of active zones, detected by Bruchpilot protein puncta distribution, remained unchanged; however, the average number of active zones per bouton decreased. These disturbances were accompanied by a decrease in frequency of miniature excitatory junction potentials without alteration in random nature of spontaneous quantal release. Similar structural and functional changes were observed with co-overexpression of human APP and β-secretase genes. In Drosophila line with expression of human amyloid-β42 peptide itself, parameters analyzed did not differ from controls, suggesting the specificity of APP effects. These results confirm the involvement of APP in synaptogenesis and provide evidence to suggest that human APP overexpression specifically disturbs the structural and functional organization of active zone and results in altered Bruchpilot distribution and lowered probability of spontaneous neurotransmitter release.

Left ventricular vascular and metabolic adaptations to high‐intensity interval and moderate intensity continuous training: a randomized trial in healthy middle‐aged men

High-intensity interval training (HIIT) has become popular, time-sparing alternative to moderate intensity continuous training (MICT), although the cardiac vascular and metabolic effects of HIIT are incompletely known. We compared the effects of 2-week interventions with HIIT and MICT on myocardial perfusion and free fatty acid and glucose uptake. Insulin-stimulated myocardial glucose uptake was decreased by training without any significantly different response between the groups, whereas free fatty acid uptake remained unchanged. Adenosine-stimulated myocardial perfusion responded differently to the training modes (change in mean HIIT: -19%; MICT: +9%; P=0.03 for interaction) and was correlated with myocardial glucose uptake for the entire dataset and especially after HIIT training. HIIT and MICT induce similar metabolic and functional changes in the heart, although myocardial vascular hyperaemic reactivity is impaired after HIIT, and this should be considered when prescribing very intense HIIT for previously untrained subjects. High-intensity interval training (HIIT) is a time-efficient way of obtaining the health benefits of exercise, although the cardiac effects of this training mode are incompletely known. We compared the effects of short-term HIIT and moderate intensity continuous training (MICT) interventions on myocardial perfusion and metabolism and cardiac function in healthy, sedentary, middle-aged men. Twenty-eight healthy, middle-aged men were randomized to either HIIT or MICT groups (n=14 in both) and underwent six cycle ergometer training sessions within 2weeks (HIIT session: 4-6×30s all-out cycling/4min recovery, MICT session 40-60min at 60% V̇O2 peak ). Cardiac magnetic resonance imaging (CMRI) was performed to measure cardiac structure and function and positron emission tomography was used to measure myocardial perfusion at baseline and during adenosine stimulation, insulin-stimulated glucose uptake (MGU) and fasting free fatty acid uptake (MFFAU). End-diastolic and end-systolic volumes increased and ejection fraction slightly decreased with both training modes, although no other changes in CMRI were observed. MFFAU and basal myocardial perfusion remained unchanged. MGU was decreased by training (HIIT from 46.5 to 35.9; MICT from 47.4 to 44.4mmol100g-1 min-1 , P=0.007 for time, P=0.11 for group×time). Adenosine-stimulated myocardial perfusion responded differently to the training modes (change in mean HIIT: -19%; MICT: +9%; P=0.03 for group×time interaction). HIIT and MICT induce similar metabolic and functional changes in the heart, although myocardial vascular hyperaemic reactivity is impaired after HIIT. This should be taken into account when prescribing very intense HIIT for previously untrained subjects.

Behavioural and functional vestibular disorders after space flight: 2. Fish, amphibians and birds

The review presents data on functional changes in fish, amphibians and birds associated with otolith organ activity after exposure to weightlessness during spaceflight. These data are of importance both for solving some fundamental problems of vestibulology and for practice. In the latter case, lower vertebrates are considered as a convenient and, most importantly, adequate model to unravel the mechanisms of vestibular disorders in humans. Analysis of the experimental results shows that weightlessness exerts no substantial effect on the formation and functional state of the otolith system in embryos of fish, amphibians and birds developing during spaceflight. Moreover, they even promote faster embryonic development of fish and amphibians as shown for mammalian fetuses. The experiments show that both in lower and higher vertebrates weightlessness brings about similar functional and behavioral changes. For example, in fish hatchlings and amphibian tadpoles (without lordosis) the vestibulo-ocular reflex was more pronounced immediately after orbital spaceflight than in control. An analogous alteration in the otolith reflex was observed in most cosmonauts after short-time space missions. In adult terrestrial vertebrates, as well as in humans, immediately after landing there was found a drop in the level of activity and deterioration of the equilibrium function and motor coordination. Another interesting finding was an unusual looping behavior when fish and tadpoles swam in loops post landing. Presumably, unusual motor activity of animals, as well as illusions arising in cosmonauts and astronauts during the transition from 1 to 0 g, have the same background being associated with changes in the stimulation pattern of the otolith organs. Considering the similarity of vestibular responses, the use of animal models seems very promising as allowing different invasive techniques.