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We developed a high-speed two-photon optical ribbon imaging system, which combines galvo-mirrors for an arbitrary curve scan on a lateral plane and a tunable acoustic gradient-index lens for a 100 kHz-1 MHz axial scan. The system provides micrometer/millisecond spatiotemporal resolutions, which enable continuous readout of functional dynamics from small and densely packed neurons in a living adult Drosophila brain. Compared to sparse sampling techniques, the ribbon imaging modality avoids motion artifacts. Combined with a Drosophila anatomical connectome database, which is the most complete among all model animals, this technique paves the way toward establishing whole-brain functional connectome.
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Volume imaging based on a fast focus-tunable lens (FTL) allows three-dimensional (3D) observation within milliseconds by extending the depth-of-field (DOF) with sub-micrometer transverse resolution on optical sectioning microscopes. However, the previously published DOF extensions were neither axially uniform nor fit with theoretical prediction. In this work, complete theoretical treatments of focus extension with confocal and various multiphoton microscopes are established to correctly explain the previous results. Moreover, by correctly placing the FTL and properly adjusting incident beam diameter, a uniform DOF is achieved in which the actual extension nicely agrees with the theory. Our work not only provides a theoretical platform for volumetric imaging with FTL but also demonstrates the optimized imaging condition.
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During the last decades, several resolution enhancement methods for optical microscopy beyond diffraction limit have been developed. Nevertheless, those hardware-based techniques typically require strong illumination, and fail to improve resolution in deep tissue. Here we develop a high-speed computational approach, three-dimensional virtual spatial overlap modulation microscopy (3D-vSPOM), which immediately solves the strong-illumination issue. By amplifying only the spatial frequency component corresponding to the un-scattered point-spread-function at focus, plus 3D nonlinear value selection, 3D-vSPOM shows significant resolution enhancement in deep tissue. Since no iteration is required, 3D-vSPOM is much faster than iterative deconvolution. Compared to non-iterative deconvolution, 3D-vSPOM does not need a priori information of point-spread-function at deep tissue, and provides much better resolution enhancement plus greatly improved noise-immune response. This method is ready to be amalgamated with two-photon microscopy or other laser scanning microscopy to enhance deep-tissue resolution.
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Successful management of obesity can be challenging if individuals constantly experience cravings. The present study investigated the effects of a high-protein diet, including a high-protein drink consumed immediately after high-intensity interval training (HIIT), on appetite and weight loss in obese middle-aged individuals. A total of 52 obese middle-aged individuals (58.2 ± 4.11 years old) were randomly assigned to one of three groups: the exercise group (E, n=19), exercise and high-protein diet group (ED, n=21), and a control group (n=12). The E and ED groups engaged in cycling HIIT (comprising 90 % of peak heart rate (HRpeak) for 3 min, followed by 70 % of HRpeak for 3 min, for a total of 5 cycles) three times a week for 3 months. The ED group consumed a high-protein drink immediately after HIIT and had a daily protein intake of 1.6g/kg. Body composition and eating behavior were assessed before and after the intervention. Additionally, appetite levels were measured before and after each exercise session, before dinner, and before bedtime during three phases of the intervention: the first phase (weeks 3-4), the second phase (weeks 5-8), and the third phase (weeks 9-12). Results showed that only the ED group experienced a decrease in body mass index (from 27.4 ± 4.28 to 26.8 ± 4.09 kg/m2, p=0.04). Appetite significantly increased after exercise in both E and ED groups (p values for the three phases ranged from 0.04 to 0.001 for the E group and from 0.042 to 0.003 for the ED group). The desire to eat significantly increased after exercise in the E group (phase 1: p = 0.026; phase 2: p = 0.011; phase 3: p = 0.003), but not in the ED group. Furthermore, the frequency of late-night snacking decreased in the ED group (the score changed from 2.4 ± 0.86 to 2.7 ± 0.80, p = 0.034). Notably, the E group tended to have a higher pre-dinner appetite score than the ED group in the third phase (p = 0.063). In summary, a high daily protein intake, combined with the consumption of high-protein drinks after exercise, resulted in reduced post-exercise appetite and a decrease in the frequency of late-night snacking.
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Apetite , Obesidade , Humanos , Pessoa de Meia-Idade , Apetite/fisiologia , Índice de Massa Corporal , Proteínas Alimentares , Comportamento AlimentarRESUMO
Three-dimensional printing is a non-conventional additive manufacturing process. It is different from the conventional subtractive manufacturing process. It offers exceptional rapid prototyping capabilities and results that conventional subtractive manufacturing methods cannot attain, especially in applications involving curved or intricately shaped components. Despite its advantages, metal 3D printing will face porosity, warpage, and surface roughness issues. These issues will affect the future practical application of the parts indirectly, for example, by affecting the structural strength and the parts' assembly capability. Therefore, this study compares the qualities of the warpage, weight, and surface roughness after milling and grinding processes for the same material (316L stainless steel) between rolled steel and 3D-printed steel. The experimental results show that 3D-printed parts are approximately 13% to 14% lighter than rolled steel. The surface roughness performance of 3D-printed steel is better than that of rolled steel for the same material after milling or grinding processing. The hardness of the 3D-printed steel is better than that of the rolled steel. This research verifies that 3D additive manufacturing can use surface processing to optimize surface performance and achieve the functions of lightness and hardness.
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Background: Obesity is the main risk factor of cardiovascular diseases (CVD) and metabolic diseases. The middle-aged population is the age group with the highest prevalence of obesity. Thus, improving cardiometabolic health is important to prevent CVD and metabolic diseases in middle-aged obese adults. The aim of this study was to examine the effects of exercise alone or in combination with a high-protein diet on markers of cardiometabolic health in middle-aged adults with obesity. Methods: Sixty-nine middle-aged adults with obesity were assigned randomly to the control group (C; n = 23), exercise group (E; n = 23), or exercise combined with high-protein diet group (EP; n = 23). Individuals in the E and EP groups received supervised exercise training and individuals in the EP group received high-protein diet intervention. Body composition (assessed by dual-energy X-ray absorptiometry), oral glucose tolerance test (OGTT), lipid profiles, and inflammatory markers were determined before and after 12 weeks of intervention. Insulin sensitivity index (ISI0,120) was calculated from values of fasting and 2-h insulin and glucose concentration of OGTT. Insulin-peak-time during the OGTT was recorded to reflect ß-cell function. Analysis of covariance with baseline values as covariates was used to examine the effects of the intervention. The significant level was set at 0.05. Results: After 12 weeks of intervention, the E group had a greater percentage of individuals with early insulin-peak-time during the OGTT than that in the C and EP groups (p = 0.031). EP group had lower total cholesterol and triglycerides than that in the C group (p = 0.046 and 0.014, respectively). Within-group comparisons showed that the 2-h glucose of OGTT and C-reactive protein decreased in the EP group (p = 0.013 and 0.008, respectively) but not in the E and C groups; insulin sensitivity improved in the EP group (p = 0.016) and had a trend to improve in the E group (p = 0.052); and abdominal fat mass and total body fat mass decreased in both intervention groups (p < 0.05). Conclusion: Combined high-protein diet and exercise intervention significantly decreased fat mass and improved lipid profiles, insulin sensitivity, glucose tolerance, and inflammation in middle-aged adults with obesity. Clinical Trial Registration: Thai Clinical Trials Registry, TCTR20180913003, 13-09-2018.
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Studying neural connections and activities in vivo is fundamental to understanding brain functions. Given the cm-size brain and three-dimensional neural circuit dynamics, deep-tissue, high-speed volumetric imaging is highly desirable for brain study. With sub-micrometer spatial resolution, intrinsic optical sectioning, and deep-tissue penetration capability, two-photon microscopy (2PM) has found a niche in neuroscience. However, the current 2PM typically relies on a slow axial scan for volumetric imaging, and the maximal penetration depth is only about 1â mm. Here, we demonstrate that by integrating a gradient-index (GRIN) lens and a tunable acoustic GRIN (TAG) lens into 2PM, both penetration depth and volume-imaging rate can be significantly improved. Specifically, an â¼ 1-cm long GRIN lens allows imaging relay from any target region of a mouse brain, while a TAG lens provides a sub-second volume rate via a 100 kHz â¼ 1â MHz axial scan. This technique enables the study of calcium dynamics in cm-deep brain regions with sub-cellular and sub-second spatiotemporal resolution, paving the way for interrogating deep-brain functional connectome.
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People with sarcopenic obesity (SO) are characterized by both low muscle mass (sarcopenia) and high body fat (obesity); they have greater risks of metabolic diseases and physical disability than people with sarcopenia or obesity alone. Exercise and nutrition have been reported to be effective for both obesity and sarcopenia management. Thus, we aimed to investigate the effects of exercise and nutrition on body composition, metabolic health, and physical performance in individuals with SO. Studies investigating the effects of exercise and nutrition on body composition, metabolic health, and physical performance in SO individuals were searched from electronic databases up to April 2019. Fifteen studies were included in the meta-analysis. Aerobic exercise decreased body weight and fat mass (FM). Resistance exercise (RE) decreased FM and improved grip strength. The combination of aerobic exercise and RE decreased FM and improved walking speed. Nutritional intervention, especially low-calorie high-protein (LCHP) diet, decreased FM but did not affect muscle mass and grip strength. In addition to exercise training, nutrition did not provide extra benefits in outcome. Exercise, especially RE, is essential to improve body composition and physical performance in individuals with SO. Nutritional intervention with LCHP decreases FM but does not improve physical performance.
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Dieta Redutora/métodos , Terapia por Exercício/métodos , Obesidade/terapia , Sarcopenia/terapia , Adulto , Composição Corporal , Terapia Combinada , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Força Muscular , Músculo Esquelético/fisiopatologia , Estado Nutricional , Obesidade/complicações , Obesidade/fisiopatologia , Desempenho Físico Funcional , Sarcopenia/etiologia , Sarcopenia/fisiopatologia , Resultado do TratamentoRESUMO
Drosophila is widely used in connectome studies due to its small brain size, sophisticated genetic tools, and the most complete single-neuron-based anatomical brain map. Surprisingly, even the brain thickness is only 200-µm, common Ti:sapphire-based two-photon excitation cannot penetrate, possibly due to light aberration/scattering of trachea. Here we quantitatively characterized scattering and light distortion of trachea-filled tissues, and found that trachea-induced light distortion dominates at long wavelength by comparing one-photon (488-nm), two-photon (920-nm), and three-photon (1300-nm) excitations. Whole-Drosophila-brain imaging is achieved by reducing tracheal light aberration/scattering via brain-degassing or long-wavelength excitation at 1300-nm. Our work paves the way toward constructing whole-brain connectome in a living Drosophila.
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Recently, many super-resolution technologies have been demonstrated, significantly affecting biological studies by observation of cellular structures down to nanometer precision. However, current super-resolution techniques mostly rely on wavefront engineering or wide-field imaging of signal blinking or fluctuation, and thus imaging depths are limited due to tissue scattering or aberration. Here we present a technique that is capable of imaging through an intact Drosophila brain with 20-nm lateral resolution at â¼200 µm depth. The spatial resolution is provided by molecular localization of a photoconvertible fluorescent protein Kaede, whose red form is found to exhibit blinking state. The deep-tissue observation is enabled by optical sectioning of spinning disk microscopy, as well as reduced scattering from optical clearing. Together these techniques are readily available for many biologists, providing three-dimensional resolution of densely entangled dendritic fibers in a complete Drosophila brain. The method paves the way toward whole-brain neural network studies and is applicable to other high-resolution bioimaging.
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All-optical physiology (AOP) manipulates and reports neuronal activities with light, allowing for interrogation of neuronal functional connections with high spatiotemporal resolution. However, contemporary high-speed AOP platforms are limited to single-depth or discrete multi-plane recordings that are not suitable for studying functional connections among densely packed small neurons, such as neurons in Drosophila brains. Here, we constructed a 3D AOP platform by incorporating single-photon point stimulation and two-photon high-speed volumetric recordings with a tunable acoustic gradient-index (TAG) lens. We demonstrated the platform effectiveness by studying the anterior visual pathway (AVP) of Drosophila. We achieved functional observation of spatiotemporal coding and the strengths of calcium-sensitive connections between anterior optic tubercle (AOTU) sub-compartments and >70 tightly assembled 2-µm bulb (BU) microglomeruli in 3D coordinates with a single trial. Our work aids the establishment of in vivo 3D functional connectomes in neuron-dense brain areas.