Learning cellular morphology with neural networks

Reconstruction and annotation of volume electron microscopy data sets of brain tissue is challenging but can reveal invaluable information about neuronal circuits. Significant progress has recently been made in automated neuron reconstruction as well as automated detection of synapses. However, methods for automating the morphological analysis of nanometer-resolution reconstructions are less established, despite the diversity of possible applications. Here, we introduce cellular morphology neural networks (CMNs), based on multi-view projections sampled from automatically reconstructed cellular fragments of arbitrary size and shape. Using unsupervised training, we infer morphology embeddings (Neuron2vec) of neuron reconstructions and train CMNs to identify glia cells in a supervised classification paradigm, which are then used to resolve neuron reconstruction errors. Finally, we demonstrate that CMNs can be used to identify subcellular compartments and the cell types of neuron reconstructions

Resolved Kinematics of Runaway and Field OB Stars in the Small Magellanic Cloud

We use GAIA DR2 proper motions of the RIOTS4 field OB stars in the Small Magellanic Cloud (SMC) to study the kinematics of runaway stars. The data reveal that the SMC Wing has a systemic peculiar motion relative to the SMC Bar of (v_RA, v_Dec) = (62 +/-7, -18+/-5) km/s and relative radial velocity +4.5 +/- 5.0 km/s. This unambiguously demonstrates that these two regions are kinematically distinct: the Wing is moving away from the Bar, and towards the Large Magellanic Cloud with a 3-D velocity of 64 +/- 10 km/s. This is consistent with models for a recent, direct collision between the Clouds. We present transverse velocity distributions for our field OB stars, confirming that unbound runaways comprise on the order of half our sample, possibly more. Using eclipsing binaries and double-lined spectroscopic binaries as tracers of dynamically ejected runaways, and high-mass X-ray binaries (HMXBs) as tracers of runaways accelerated by supernova kicks, we find significant contributions from both populations. The data suggest that HMXBs have lower velocity dispersion relative to dynamically ejected binaries, consistent with the former corresponding to less energetic supernova kicks that failed to unbind the components. Evidence suggests that our fast runaways are dominated by dynamical, rather than supernova, ejections.Comment: Accepted to ApJ Letters. 10 pages, 4 figure

Advanced Glycation End Products Acutely Impair Ca2+ Signaling in Bovine Aortic Endothelial Cells

Post-translational modification of proteins in diabetes, including formation of advanced glycation end products (AGEs) are believed to contribute to vascular dysfunction and disease. Impaired function of the endothelium is an early indicator of vascular dysfunction in diabetes and as many endothelial cell processes are dependent upon intracellular [Ca2+] and Ca2+ signalling, the aim of this study was to examine the acute effects of AGEs on Ca2+ signalling in bovine aortic endothelial cells (BAEC). Ca2+ signalling was studied using the fluorescent indicator dye Fura2-AM. AGEs were generated by incubating bovine serum albumin with 0 - 250 mM glucose or glucose-6-phosphate for 0 to 120 days at 37ºC. Under all conditions, the main AGE species generated was carboxymethyl lysine (CML) as assayed using both GC-MS and HPLC. In Ca2+-replete solution, exposure of BAEC to AGEs for 5 min caused an elevation in basal [Ca2+] and attenuated the increase in intracellular [Ca2+] caused by ATP (100 µM). In the absence of extracellular Ca2+, exposure of BAEC to AGEs for 5 min caused an elevation in basal [Ca2+] and attenuated subsequent intracellular Ca2+ release caused by ATP, thapsigargin (0.1 µM) and ionomycin (3 µM), but AGEs did not affect extracellular Ca2+ entry induced by the re-addition of Ca2+ to the bathing solution in the presence of any of these agents. The anti-oxidant α-lipoic acid (2 µM) and NAD(P)H oxidase inhibitors apocynin (500 µM) and diphenyleneiodonium (DPI, 1 µM) abolished these effects of AGEs on BAECs, as did the IP3 receptor antagonist xestospongin C (1 µM). In summary, AGEs caused an acute depletion of Ca2+ from the intracellular store in BAECs, such that the Ca2+ signal stimulated by the subsequent application other agents acting upon this store is reduced. The mechanism may involve generation of ROS from NAD(P)H oxidase and possible activation of the IP3 receptor

HbA1c variability in adults with type 1 diabetes on continuous subcutaneous insulin infusion (CSII) therapy compared to multiple daily injection (MDI) treatment

Objective To determine if continuous subcutaneous insulin infusion (CSII) therapy is associated with lower glycated haemoglobin (HbA1c) variability (long-term glycaemic variability; GV) relative to multiple daily injection (MDI) treatment in adults with type 1 diabetes mellitus (T1DM). Design Retrospective audit. Setting and participants Clinic records from 506 adults with T1DM from two tertiary Australian hospitals. Outcome measures Long-term GV was assessed by HbA1c SD and coefficient of variation (CV) in adults on established MDI or CSII therapy, and in a subset changing from MDI to CSII. Results Adults (n=506, (164 CSII), 50% women, mean±SD age 38.0±15.3 years, 17.0±13.7 years diabetes, mean HbA1c 7.8%±1.2% (62±13 mmol/mol) on CSII, 8.0%±1.5% (64±16 mmol/mol) on MDI) were followed for 4.1±3.6 years. CSII use was associated with lower GV (HbA1c SD: CSII vs MDI 0.5%±0.41% (6±6 mmol/mol) vs 0.7%±0.7% (9±8 mmol/mol)) and CV: CSII vs MDI 6.7%±4.6% (10±10 mmol/mol) vs 9.3%±7.3% (14±13 mmol/mol), both p<0.001. Fifty-six adults (73% female, age 36±13 years, 16±13 years diabetes, HbA1c 7.8%±0.8% (62±9 mmol/mol)) transitioned from MDI to CSII. Mean HbA1c fell by 0.4%. GV from 1 year post-CSII commencement decreased significantly, HbA1c SD pre-CSII versus post-CSII 0.7%±0.5% (8±5 mmol/mol) vs 0.4%±0.4% (5±4 mmol/mol); p<0.001, and HbA1c CV 9.2%±5.5% (13±8 mmol/mol) vs 6.1%±3.9% (9±5 mmol/mol); p<0.001. Conclusions In clinical practice with T1DM adults relative to MDI, CSII therapy is associated with lower HbA1c GV

Stationarity, soft ergodicity, and entropy in relativistic systems

Recent molecular dynamics simulations show that a dilute relativistic gas equilibrates to a Juettner velocity distribution if ensemble velocities are measured simultaneously in the observer frame. The analysis of relativistic Brownian motion processes, on the other hand, implies that stationary one-particle distributions can differ depending on the underlying time-parameterizations. Using molecular dynamics simulations, we demonstrate how this relativistic phenomenon can be understood within a deterministic model system. We show that, depending on the time-parameterization, one can distinguish different types of soft ergodicity on the level of the one-particle distributions. Our analysis further reveals a close connection between time parameters and entropy in special relativity. A combination of different time-parameterizations can potentially be useful in simulations that combine molecular dynamics algorithms with randomized particle creation, annihilation, or decay processes.Comment: 4 page

An intimate collaboration between peroxisomes and lipid bodies

Although peroxisomes oxidize lipids, the metabolism of lipid bodies and peroxisomes is thought to be largely uncoupled from one another. In this study, using oleic acid–cultured Saccharomyces cerevisiae as a model system, we provide evidence that lipid bodies and peroxisomes have a close physiological relationship. Peroxisomes adhere stably to lipid bodies, and they can even extend processes into lipid body cores. Biochemical experiments and proteomic analysis of the purified lipid bodies suggest that these processes are limited to enzymes of fatty acid β oxidation. Peroxisomes that are unable to oxidize fatty acids promote novel structures within lipid bodies (“gnarls”), which may be organized arrays of accumulated free fatty acids. However, gnarls are suppressed, and fatty acids are not accumulated in the absence of peroxisomal membranes. Our results suggest that the extensive physical contact between peroxisomes and lipid bodies promotes the coupling of lipolysis within lipid bodies with peroxisomal fatty acid oxidation

Continuous subcutaneous insulin infusion alters microRNA expression and glycaemic variability in children with type 1 diabetes

To determine whether continuous subcutaneous insulin infusion (CSII) vs. multiple daily injections (MDI) therapy from near-diagnosis of type 1 diabetes is associated with reduced glycaemic variability (GV) and altered microRNA (miRNAs) expression. Adolescents (74% male) within 3-months of diabetes diagnosis (n = 27) were randomized to CSII (n = 12) or MDI. HbA1c, 1-5-Anhydroglucitol (1,5-AG), high sensitivity C-peptide and a custom TaqMan qPCR panel of 52 miRNAs were measured at baseline and follow-up (median (LQ-UQ); 535 (519-563) days). There were no significant differences between groups in baseline or follow-up HbA1c or C-peptide, nor baseline miRNAs. Mean +/- SD 1,5-AG improved with CSII vs. MDI (3.1 +/- 4.1 vs. - 2.2 +/- - 7.0 mg/ml respectively, P = 0.029). On follow-up 11 miRNAs associated with diabetes vascular complications had altered expression in CSII-users. Early CSII vs. MDI use is associated with lower GV and less adverse vascular-related miRNAs. Relationships with future complications are of interest

Increased serum kallistatin levels in type 1 diabetes patients with vascular complications

BACKGROUND: Kallistatin, a serpin widely produced throughout the body, has vasodilatory, anti-angiogenic, anti-oxidant, and anti-inflammatory effects. Effects of diabetes and its vascular complications on serum kallistatin levels are unknown. METHODS: Serum kallistatin was quantified by ELISA in a cross-sectional study of 116 Type 1 diabetic patients (including 50 with and 66 without complications) and 29 non-diabetic controls, and related to clinical status and measures of oxidative stress and inflammation. RESULTS: Kallistatin levels (mean(SD)) were increased in diabetic vs. control subjects (12.6(4.2) vs. 10.3(2.8) μg/ml, p = 0.007), and differed between diabetic patients with complications (13.4(4.9) μg/ml), complication-free patients (12.1(3.7) μg/ml), and controls; ANOVA, p = 0.007. Levels were higher in diabetic patients with complications vs. controls, p = 0.01, but did not differ between complication-free diabetic patients and controls, p > 0.05. On univariate analyses, in diabetes, kallistatin correlated with renal dysfunction (cystatin C, r = 0.28, p = 0.004; urinary albumin/creatinine, r = 0.34, p = 0.001; serum creatinine, r = 0.23, p = 0.01; serum urea, r = 0.33, p = 0.001; GFR, r = -0.25, p = 0.009), total cholesterol (r = 0.28, p = 0.004); LDL-cholesterol (r = 0.21, p = 0.03); gamma-glutamyltransferase (GGT) (r = 0.27, p = 0.04), and small artery elasticity, r = -0.23, p = 0.02, but not with HbA1c, other lipids, oxidative stress or inflammation. In diabetes, geometric mean (95%CI) kallistatin levels adjusted for covariates, including renal dysfunction, were higher in those with vs. without hypertension (13.6 (12.3-14.9) vs. 11.8 (10.5-13.0) μg/ml, p = 0.03). Statistically independent determinants of kallistatin levels in diabetes were age, serum urea, total cholesterol, SAE and GGT, adjusted r2 = 0.24, p < 0.00001. CONCLUSIONS: Serum kallistatin levels are increased in Type 1 diabetic patients with microvascular complications and with hypertension, and correlate with renal and vascular dysfunction

Cardiometabolic risk factors, peripheral arterial tonometry and metformin in adults with type 1 diabetes participating in the REducing with MetfOrmin Vascular Adverse Lesions trial

BACKGROUND: Peripheral arterial tonometry (PAT) provides non-invasive measures of vascular health. Beneficial effects of metformin on vascular function have been reported in youth with type 1 diabetes (T1D). In the REducing with MetfOrmin Vascular Adverse Lesions (REMOVAL) trial in adults with T1D and high cardiovascular risk, we examined: (i) the extent to which routinely-measured cardiometabolic risk factors explain variance in baseline PAT; and (ii) the effects of metformin on PAT measures. METHODS: Cross-sectional univariable and multivariable analyses of baseline reactive hyperaemia index (RHI) and augmentation index (AI) (EndoPAT® (Itamar, Israel); and analysis of 36-months metformin versus placebo on vascular tonometry. RESULTS: In 364 adults ((mean ± SD) age 55.2 ± 8.5 years, T1D 34.0 ± 10.6 years, HbA1c 64.5 ± 9.0 mmol/mol (8.1 ± 0.8%)), RHI was 2.26 ± 0.74 and AI was 15.9 ± 19.2%. In an exhaustive search, independent associates of (i) RHI were smoking, waist circumference, systolic blood pressure and vitamin B12 (adjusted R2 = 0.11) and (ii) AI were male sex, pulse pressure, heart rate and waist circumference (adjusted R2 = 0.31). Metformin did not significantly affect RHI or AI. CONCLUSION: Cardiometabolic risk factors explained only a modest proportion of variance in PAT measures of vascular health in adults with T1D and high cardiovascular risk. PAT measures were not affected by metformin