We demonstrate residual channel attention networks (RCAN) for restoring and enhancing volumetric time-lapse (4D) fluorescence microscopy data. First, we modify RCAN to handle image volumes, showing that our network enables denoising competitive with three other state-of-the-art neural networks. We use RCAN to restore noisy 4D super-resolution data, enabling image capture over tens of thousands of images (thousands of volumes) without apparent photobleaching. Second, using simulations we show that RCAN enables class-leading resolution enhancement, superior to other networks. Third, we exploit RCAN for denoising and resolution improvement in confocal microscopy, enabling ~2.5-fold lateral resolution enhancement using stimulated emission depletion (STED) microscopy ground truth. Fourth, we develop methods to improve spatial resolution in structured illumination microscopy using expansion microscopy ground truth, achieving improvements of ~1.4-fold laterally and ~3.4-fold axially. Finally, we characterize the limits of denoising and resolution enhancement, suggesting practical benchmarks for evaluating and further enhancing network performance.data, which we deconvolved to yield high SNR 'ground truth'. We then used 30 of these volumes for training and held out volumes for testing network performance. Using the same training and test data, we compared four networks: RCAN, CARE, SRResNET 20 , and ESRGAN 21 . SRResNet and ESRGAN are both class-leading deep residual networks used in image super-resolution, with ESRGAN winning the 2018 Perceptual Image Restoration and Manipulation challenge on perceptual image super-resolution 22 .For the mEmerald-Tomm20 label, RCAN, CARE, ESRGAN, and SRResNET predictions all provided 88 clear improvements in visual appearance, structural similarity index (SSIM) and peak signal-to-noise-89 ratio (PSNR) metrics relative to the raw input (Fig. 1b), also outperforming direct deconvolution on the noisy input data (Supplementary Fig. 1). The RCAN output provided PSNR and SSIM values competitive with the other networks (Fig. 1b), prompting us to investigate whether this performance held for other organelles. We thus conducted similar experiments for fixed U2OS cells with labeled actin, endoplasmic reticulum (ER), golgi, lysosomes, and microtubules (Supplementary Fig. 2), acquiring 15-23 volumes of training data and training independent networks for each organelle. In almost all cases, RCAN performance met or exceeded the other networks (Supplementary Fig. 3, Supplementary Table 3).An essential consideration when using any deep learning method is understanding when network performance deteriorates. Independently training an ensemble of networks and computing measures of network disagreement can provide insight into this issue 9,16 , yet such measures were not generally predictive of disagreement between ground truth and RCAN output (Supplementary Fig. 4). Instead, we found that estimating the per-pixel SNR in the raw input (Methods, Supplementary Fig. 4) seemed to better correlate with network ...

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. Enhancement of whole body glucose uptake during and after human skeletal muscle low-frequency electrical stimulation. J Appl Physiol 94: 2107-2112, 2003. First published January 31, 2003 10.1152/japplphysiol.00486.2002There is considerable evidence to suggest that electrical stimulation (ES) activates glucose uptake in rodent skeletal muscle. It is, however, unknown whether ES can lead to similar metabolic enhancement in humans. We employed low-frequency ES through surface electrodes placed over motor points of quadriceps femoris muscles. In male subjects lying in the supine position, the highest oxygen uptake was obtained by a stimulation pattern with 0.2-ms biphasic square pulses at 20 Hz and a 1-s on-off duty cycle. Oxygen uptake was increased by approximately twofold throughout the 20-min stimulation period and returned to baseline immediately after stimulation. Concurrent elevation of the respiratory exchange ratio and blood lactate concentration indicated anaerobic glycogen breakdown and utilization during ES. Whole body glucose uptake determined by the glucose disposal rate during euglycemic clamp was acutely increased by 2.5 mg ⅐ kg Ϫ1 ⅐ min Ϫ1 in response to ES and, moreover, remained elevated by 3-4 mg ⅐ kg Ϫ1 ⅐ min Ϫ1 for at least 90 min after cessation of stimulation. Thus the stimulatory effect of ES on whole body glucose uptake persisted not only during, but also after, stimulation. Low-frequency ES may become a useful therapeutic approach to activate energy and glucose metabolism in humans. glucose transport; euglycemic clamp; exercise; insulin sensitivity; oxygen uptake PHYSICAL EXERCISE HAS PROFOUND effects on energy and fuel metabolism in contracting skeletal muscle. It is well established that exercise can directly activate glucose uptake in skeletal muscle by inducing translocation of GLUT-4 glucose transporters to the cell surface via an insulin-independent mechanism (contraction-stimulated glucose transport) (10,11,31,37,49). This phenomenon is considered responsible for the acute effect of exercise on glucose transport, with the majority of glucose being taken up by contracting skeletal muscle. In fact, contraction-stimulated GLUT-4 translocation is not impaired in insulin-resistant conditions such as Type 2 diabetes and obesity (25). Furthermore, the period after exercise is also characterized by a substantial increase in insulin sensitivity that leads to insulin-dependent GLUT-4 translocation and glucose transport as a local phenomenon restricted to exercised muscles (9, 13, 39, 40, 48a). Thus these insulin-independent and -dependent mechanisms of exercise have been widely utilized to prevent individuals from developing glucose intolerance and to improve glycemic control in patients with Type 2 diabetes.Clinically, electrical stimulation (ES) of muscle is useful as a modality of assisting muscle contraction for those who have difficulties in performing voluntary exercise. The use of ES has been traditionally employed for muscle strengthening, maintenance of muscle mass and strength, a...

We report a retrospective analysis of children with myelodysplastic syndrome (MDS) diagnosed between 1990 and 1997 in Japan. In total, 189 patients were enrolled: 122 cases of primary MDS (26 RA, 18 RAEB, 25 RAEBt, 53 CMML/JMML), 24 cases with constitutional predisposition to MDS, and 43 cases of therapy-related MDS (t-MDS). The frequency of pediatric MDS was estimated to be 7.7% of all leukemias. Cytogenetic abnormalities were observed in 41% of primary MDS and 90% of t-MDS cases. The 4-year survival rate, estimated by Kaplan-Meier analysis, for primary RA was 78.9%, while other types of MDS and JMML had rates lower than 40%, and t-MDS showed an even more unfavorable prognosis. In primary MDS, the survival rate of patients with cytogenetic abnormalities was significantly lower. Among prognostic variables by IPSS, only the cytogenetic pattern was useful for predicting outcome in childhood MDS. There was no apparent advantage to chemotherapy for RA, and the survival rate in patients with primary RA, JMML, or t-MDS receiving stem cell transplantation was significantly higher. More precise designs of our diagnostic and classification systems, as well as therapeutic trials in large-scale prospective studies, are necessary for further improvements in MDS outcome. Leukemia (2001) 15, 1713-1720.

The improved outcome of acquired aplastic anemia (AA) has revealed later complications, such as myelodysplastic syndrome (MDS) and acute myelogenous leukemia (AML). We retrospectively analyzed 167 children with severe acquired AA. Eleven of 50 children treated with cyclosporin (CSA) and recombinant human granulocyte colony-stimulating factor (rhG-CSF ) developed MDS/AML; 8 of these were within 36 months of the diagnosis of AA, much earlier than previous reports. Six of the 11 children received rhG-CSF exceeding 10 μg/kg/d, and 9 received rhG-CSF therapy for over 1 year. Ten children showed monosomy 7 at diagnosis of MDS. All of the 11 children were administered both CSA and rhG-CSF. There was no development of MDS/AML among 41 children treated with either CSA or rhG-CSF or among 48 children who underwent bone marrow transplantation. A well-controlled clinical trial is warranted to determine whether therapeutic modalities affect the development of MDS/AML in children with severe acquired AA.

To investigate the effect of sucrose or caffeine ingestion on the performance of prolonged running, five male distance runners attending senior high school (15.6 yrs) carried out running on a treadmill at an intensity corresponding to the individuals' 80% VO2 max until exhaustion. Before and 45 min after exercise, the subjects were given either a placebo (Con), sucrose (81 +/- 18 g) (Su), caffeine (384 +/- 13 mg) (Caf), or sucrose (72 +/- 22 g) plus caffeine (396 +/- 29 mg) (Su + Caf) solution. The duration of the exercise was significantly longer in Su, Caf, and Su + Caf than in Con. The duration in four of five subjects was longest in Su + Caf, although it was not significantly different from that in Su or Caf. Carbohydrate (CHO) utilization was highest in Su while fat utilization was highest in Caf. The energy supply from both sources was almost the same between Con and Su + Caf. The plasma glucose concentration was higher in Su than in Con. The plasma free fatty acid (FFA) level was higher in Caf than in Con. The plasma glucose and lactic acid concentrations were highest in Su + Caf while the plasma FFA level was the same as in Con. In conclusion, ingestion of sucrose, caffeine, or sucrose plus caffeine solution was equally effective in improving endurance during running carried out at an intensity of approximately 80% VO2 max.

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