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1.
EMBO Rep ; 24(5): e56114, 2023 05 04.
Artigo em Inglês | MEDLINE | ID: mdl-36929726

RESUMO

Vesicular transport is a means of communication. While cells can communicate with each other via secretion of extracellular vesicles, less is known regarding organelle-to organelle communication, particularly in the case of mitochondria. Mitochondria are responsible for the production of energy and for essential metabolic pathways in the cell, as well as fundamental processes such as apoptosis and aging. Here, we show that functional mitochondria isolated from Saccharomyces cerevisiae release vesicles, independent of the fission machinery. We isolate these mitochondrial-derived vesicles (MDVs) and find that they are relatively uniform in size, of about 100 nm, and carry selective protein cargo enriched for ATP synthase subunits. Remarkably, we further find that these MDVs harbor a functional ATP synthase complex. We demonstrate that these vesicles have a membrane potential, produce ATP, and seem to fuse with naive mitochondria. Our findings reveal a possible delivery mechanism of ATP-producing vesicles, which can potentially regenerate ATP-deficient mitochondria and may participate in organelle-to-organelle communication.


Assuntos
Mitocôndrias , Saccharomyces cerevisiae , Potenciais da Membrana , Mitocôndrias/metabolismo , Transporte Biológico , Saccharomyces cerevisiae/genética , Saccharomyces cerevisiae/metabolismo , Trifosfato de Adenosina/metabolismo
2.
J Mol Cell Cardiol ; 193: 113-124, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38960316

RESUMO

The sarcolemmal Ca2+ efflux pathways, Na+-Ca2+-exchanger (NCX) and Ca2+-ATPase (PMCA), play a crucial role in the regulation of intracellular Ca2+ load and Ca2+ transient in cardiomyocytes. The distribution of these pathways between the t-tubular and surface membrane of ventricular cardiomyocytes varies between species and is not clear in human. Moreover, several studies suggest that this distribution changes during the development and heart diseases. However, the consequences of NCX and PMCA redistribution in human ventricular cardiomyocytes have not yet been elucidated. In this study, we aimed to address this point by using a mathematical model of the human ventricular myocyte incorporating t-tubules, dyadic spaces, and subsarcolemmal spaces. Effects of various combinations of t-tubular fractions of NCX and PMCA were explored, using values between 0.2 and 1 as reported in animal experiments under normal and pathological conditions. Small variations in the action potential duration (≤ 2%), but significant changes in the peak value of cytosolic Ca2+ transient (up to 17%) were observed at stimulation frequencies corresponding to the human heart rate at rest and during activity. The analysis of model results revealed that the changes in Ca2+ transient induced by redistribution of NCX and PMCA were mainly caused by alterations in Ca2+ concentrations in the subsarcolemmal spaces and cytosol during the diastolic phase of the stimulation cycle. The results suggest that redistribution of both transporters between the t-tubular and surface membranes contributes to changes in contractility in human ventricular cardiomyocytes during their development and heart disease and may promote arrhythmogenesis.


Assuntos
Cálcio , Ventrículos do Coração , Miócitos Cardíacos , Sarcolema , Trocador de Sódio e Cálcio , Humanos , Miócitos Cardíacos/metabolismo , Cálcio/metabolismo , Trocador de Sódio e Cálcio/metabolismo , Ventrículos do Coração/metabolismo , Sarcolema/metabolismo , Potenciais de Ação , Sinalização do Cálcio , Membrana Celular/metabolismo , Modelos Biológicos , Modelos Cardiovasculares
3.
J Nutr ; 154(4): 1347-1355, 2024 04.
Artigo em Inglês | MEDLINE | ID: mdl-38365118

RESUMO

BACKGROUND: Preservation of fat-free mass (FFM) during intentional weight loss is challenging yet important to maintain a resting metabolic rate. A balanced protein distribution of 25-30 g per meal improves 24-h muscle protein synthesis, which may promote FFM maintenance and greater reductions in fat mass (FM) during weight loss in women. OBJECTIVES: We aimed to determine whether the daily dietary protein distribution pattern during energy restriction influences changes in body composition in women of reproductive age. We hypothesized that evenly distributing protein across meals compared with the usual intake pattern of consuming most of the protein at the dinner meal would be superior in preserving FFM while reducing FM during weight loss. METHODS: Healthy women (n = 43) aged 20-44 y with a BMI of 28-45 kg/m2 completed a randomized parallel feeding study testing 2 patterns of daily protein intake (even distribution across all meals compared with a skewed distribution with most protein consumed at the evening meal). Participants completed an 8-wk controlled 20% energy restriction (all foods provided), followed by an 8-wk self-choice phase in which participants were asked to maintain a similar diet and dietary pattern when purchasing and consuming their own foods. Body composition was measured at baseline, week 8, and week 16. Data were analyzed using mixed models. Statistical significance was set at P < 0.05. Data are presented as differences in least squares means ± SE. RESULTS: No significant main effects of group or group-by-time interactions were observed. All measures exhibited the main effect of time (P < 0.001). Overall, body weight, FFM, FM, and body fat percentage decreased 5.6 ± 0.4, 1.0 ± 0.2, 4.6 ± 0.4 kg, and 2.3 ± 0.2%, respectively, during this 16-wk study. CONCLUSION: Daily dietary protein distribution at a fixed protein level does not appear to influence changes in body composition during weight loss in women of reproductive age. CLINICAL TRIAL REGISTRY NUMBER AND WEBSITE WHERE IT WAS OBTAINED: NCT03202069 https://classic. CLINICALTRIALS: gov/ct2/show/NCT03202069.


Assuntos
Obesidade , Sobrepeso , Humanos , Feminino , Índice de Massa Corporal , Dieta , Redução de Peso , Composição Corporal , Refeições , Proteínas Alimentares
4.
Eur J Nutr ; 63(5): 1747-1757, 2024 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-38563983

RESUMO

AIMS: Evidence regarding impact of protein intake distribution on skeletal muscle mass in older adults is limited and inconsistent. This study aims to investigate the relationship of evenness of dietary protein distribution and number of meals exceeding a threshold with appendicular skeletal muscle mass (ASM) in healthy and free-living Chinese older adults. METHODS: Repeated measured data of 5689 adult participants aged ≥ 60 years from the China Health and Nutrition Survey (CHNS) 2015 and 2018 waves were analyzed. Mixed-effects linear regression model was performed to examine the relationship between coefficient of variance (CV) of protein intake across meals, number of meals ≥ 0.4 g protein/kg BW and ASM, respectively. Analyses were conducted separately for male and female. RESULTS: The average CV of protein intake in each wave was in the range of 0.34-0.35. More than 40% male and female participants in each wave had no meal reaching 0.4 g protein/kg BW. Female participants in the highest quartile of protein intake CV had significantly lower ASM (ß = -0.18, 95%CI = -0.32, -0.04) compared with those in the lowest quartile, after adjustment for multiple confounders. Significant negative trends were observed across dietary protein CV quartiles with ASM both in male (P trend = 0.043) and female (P trend = 0.007). Significant positive association between number of meals exceeding 0.4 g protein /kg BW and relative ASM were observed in females (2 meals vs. 0 meal: ß = 0.003, 95%CI = 0.0007,0.006;≥3 meals vs. 0 meal: ß = 0.008, 95%CI = 0.003,0.013), after adjusting for multiple covariates. CONCLUSIONS: A more even-distributed protein intake pattern and more meals reaching protein intake threshold were respectively associated with higher appendicular skeletal muscle mass in healthy and free-living older Chinese adults. Prospective studies and intervention trials are needed to confirm these cross-sectional findings.


Assuntos
Proteínas Alimentares , Músculo Esquelético , Inquéritos Nutricionais , Humanos , Masculino , Feminino , Proteínas Alimentares/administração & dosagem , Idoso , Músculo Esquelético/fisiologia , Músculo Esquelético/metabolismo , China , Pessoa de Meia-Idade , Inquéritos Nutricionais/métodos , Inquéritos Nutricionais/estatística & dados numéricos , Dieta/métodos , Dieta/estatística & dados numéricos , Idoso de 80 Anos ou mais , Composição Corporal , Refeições , Estudos Transversais , População do Leste Asiático
5.
Chirality ; 36(3): e23655, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-38419363

RESUMO

This study reports the microscopic measurements of vibrational circular dichroism (VCD) on four different insect wings using a quantum cascade laser VCD system equipped with microscopic scanning capabilities (named multi-dimensional VCD [MultiD-VCD]). Wing samples, including (i) beetle, Anomala albopilosa (female), (ii) European hornet, Verspa crabro flavofasciata Cameron, 1903 (female), (iii) tiny dragonfly, Nannophya pygmae Rambur, 1842 (male), and (iv) dragonfly, Symetrum gracile Oguma, 1915 (male), were used in this study. Two-dimensional patterns of VCD signals (~10 mm × 10 mm) were obtained at a spatial resolution of 100 µm. Measurements covered the absorption peaks assigned to amides I and II in the range of 1500-1740 cm-1 . The measurements were based on the enhancement of VCD signals for the stereoregular linkage of peptide groups. The patterns were remarkably dependent on the species. In samples (i) and (ii), the wings comprised segregated domains of protein aggregates of different secondary structures. The size of each microdomain was approximately 100 µm. In contrast, no clear VCD spectra were detected in samples (iii) and (iv). One possible reason was that the chain of stereoregular polypeptides was too short to achieve VCD enhancement in samples (iii) and (iv). Notably, the unique features were only observed in the VCD spectra because the IR spectra were nearly the same among the species. The VCD results hinted at the connection of protein microscopic structures with the wing flapping mechanisms of each species.


Assuntos
Odonatos , Feminino , Masculino , Animais , Dicroísmo Circular , Estereoisomerismo , Peptídeos/química , Proteínas
6.
Cell Tissue Res ; 388(3): 503-519, 2022 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-35332371

RESUMO

The Na,K-ATPase (NKA) is an essential ion transporter and signaling molecule in all animal tissues and believed to consist at least one α and one ß-subunit to form a functional enzyme. In the large milkweed bug, Oncopeltus fasciatus, adaptation to dietary cardiac glycosides (CGs), which can fatally block the NKA, has resulted in gene duplications leading to four α1-subunits. These differ in sensitivity to CGs, but resistance trades off against ion pumping activity, thus influencing the α1-subunits' suitability for specific tissues. Besides, O. fasciatus possesses four different ß-subunits that can alter the NKA's kinetics and should play an essential role in the formation of cellular junctions.Proteomic analyses revealed the distribution and composition of α1/ß-complexes in the nervous tissue of O. fasciatus. The highly CG-resistant, but less active α1B and the highly active, but less resistant α1C predominated in the nervous tissue and co-occurred with ß2 and ß3, partly forming larger complexes than just heterodimers. Immunohistochemical analyses provided a fine scale resolution of the subunits' distribution in different morphological structures of the nervous tissue. This may suggest that α1 as well as ß-subunits occur in isolation without the other subunit, which contradicts the present understanding that the two types of subunits have to associate to form functional complexes. An isolated occurrence was especially prominent for ß3 and ßx, the enigmatic fourth and N-terminally largely truncated ß-subunit. We hypothesize that dimerization of these ß-subunits plays a role in cell-cell contacts.


Assuntos
Heterópteros , Tecido Nervoso , Animais , Duplicação Gênica , Heterópteros/metabolismo , Tecido Nervoso/metabolismo , Proteômica , ATPase Trocadora de Sódio-Potássio/metabolismo
7.
Aging Clin Exp Res ; 34(4): 927-934, 2022 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-34725773

RESUMO

BACKGROUND: Changes in nutrient intakes and protein distribution were analyzed in response to a whole food-based dietary intervention targeting high-protein meals in older adults. METHODS: Community-dwelling older adults (n = 56; M/F, 28/28; age, 69.3 ± 4.0 years) completed a 12-week intervention after randomization to exercise only (EX, n = 19), nutrition only (NUTR, n = 16), or nutrition plus exercise (NUTR + EX, n = 21). NUTR and NUTR + EX followed a dietary intervention targeting ~ 0.4 g/kg of protein at each of breakfast, lunch and dinner. RESULTS: Relative protein intake increased in NUTR (0.99 ± 0.34 to 1.43 ± 0.39 g/kg, P < 0.001) and NUTR + EX (0.90 ± 0.20 to 1.57 ± 0.49 g/kg, P < 0.001). Intakes of cholesterol, B vitamins, selenium and iodine were increased in both NUTR and NUTR + EX (P < 0.05 for all). CONCLUSION: This dietary intervention was effective at increasing daily protein intake and achieving an even distribution pattern. Changes in micronutrient intake were marked, and reflect the increase in consumption of animal-derived protein-rich food sources.


Assuntos
Ingestão de Alimentos , Refeições , Idoso , Desjejum , Dieta , Ingestão de Energia , Feminino , Humanos , Vida Independente , Masculino , Estado Nutricional
8.
Eur J Nutr ; 60(6): 3185-3202, 2021 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-33550490

RESUMO

PURPOSE: Age-related decrease in muscle mass is, among several other factors, caused by suboptimal dietary protein intake. The protein intake of the general population has a skewed distribution towards the evening meal. However, it is hypothesised that an intake of protein with an even meal distribution leads to a more frequently maximised protein synthesis. This review investigates whether an even protein distribution is associated with preservation or gain in muscle mass, muscle strength, and protein turnover. METHODS: Seven databases: PubMed, Web of Science, Google Scholar, CINAHL, Cochrane Database of Systematic Reviews, Cochrane Central Register of Controlled Trials, and Embase were searched. Studies included had a healthy population between 20 and 85 years of age, with a BMI between 18.5 and 30.0, investigated even vs. skewed protein distribution, and measured skeletal muscle relevant outcomes. Case studies and systematic reviews were excluded. Studies were appraised using the AXIS scale for observational studies and the PEDro scale for the remaining studies. RESULTS: Fifteen studies met the eligibility criteria and were included. Three out of seven studies showed an association between even protein distribution and higher muscle mass. Two out of seven studies showed an association between greater muscle strength and an even protein distribution. Only one out of six studies found a positive association between protein synthesis and an even protein distribution. CONCLUSION: Evidence indicated an association between muscle mass and an even protein intake. However, the evidence is currently insufficient to conclude whether an even protein intake is positively associated with muscle strength or protein turnover.


Assuntos
Proteínas Alimentares , Força Muscular , Adulto , Dieta , Humanos , Músculo Esquelético
9.
J Nutr ; 150(7): 1845-1851, 2020 07 01.
Artigo em Inglês | MEDLINE | ID: mdl-32321161

RESUMO

BACKGROUND: Although daily protein intake (PI) has been reported to be essential for regulating muscle mass, the distribution of daily PI in individuals is typically the lowest at breakfast and skewed toward dinner. Skewed protein intake patterns and inadequate PI at breakfast were reported to be negative factors for muscle maintenance. OBJECTIVES: This study examined whether a protein-enriched meal at breakfast is more effective for muscle accretion compared with the typical skewed PI pattern. METHODS: This 12-wk, parallel-group, randomized clinical trial included 26 men (means ± SEs; age: 20.8 ± 0.4 y; BMI: 21.8 ± 0.4 kg/m2). The "high breakfast" (HBR) group (n = 12) consumed a protein-enriched meal at breakfast providing a PI of 0.33 g/kg body weight (BW); their PI at lunch (0.46 g/kg BW) and dinner (0.48 g/kg BW) provided an adequate overall daily PI (1.30 g/kg BW/d). The "low breakfast" (LBR) group (n = 14) consumed 0.12 g protein/kg BW at breakfast; intakes at lunch (0.45 g/kg BW) and dinner (0.83 g/kg BW) yielded the same daily PI as in the HBR group. The participants performed supervised resistance training (RT) 3 times per week (75-80% 1-repetition maximum; 3 sets × 10 repetitions). DXA was used to measure the primary outcome variable, that is, total lean soft tissue mass (LTM). RESULTS: The total LTM at baseline did not differ between the HBR (52.4 ± 1.3 kg) and LBR (53.4 ± 1.2 kg) groups. After the intervention, increases in total LTM were significant in both groups, with that in the HBR group (2.5 ± 0.3 kg) tending to be greater than that in the LBR group (1.8 ± 0.3 kg) (P = 0.06), with a large effect size (Cohen d = 0.795). CONCLUSIONS: For RT-induced muscle hypertrophy in healthy young men, consuming a protein-enriched meal at breakfast and less protein at dinner while achieving an adequate overall PI is more effective than consuming more protein at dinner.This study was registered at University hospital Medical Information Network (UMIN) Clinical Trials Registry as UMIN000037583 (https://upload.umin.ac.jp/cgi-open-bin/ctr_e/ctr_view.cgi?recptno=R000042763).


Assuntos
Proteínas Alimentares/administração & dosagem , Refeições , Músculo Esquelético/crescimento & desenvolvimento , Treinamento Resistido , Adolescente , Adulto , Humanos , Masculino , Adulto Jovem
10.
J Microsc ; 278(2): 76-88, 2020 05.
Artigo em Inglês | MEDLINE | ID: mdl-32144777

RESUMO

The applicability of confocal laser scanning microscopy is limited, e.g. by attenuation of the excitation and the fluorescence emission beam. As a prerequisite for further processing and analysis of the obtained microscopic images, a new method is presented for correcting this attenuation. The correction is based on beam modelling and on a differential form of the modified Beer-Lambert law. It turns out that the intensity decay can be modelled as a double convolution of the microscopic image with the intensities of the excitation semibeam and the emission beam. Under weak assumptions made for the intensities of the fluorescent radiation and the detected signal, formulas for the attenuation correction and the attenuation simulation are derived. The method traces back to that one published by Roerdink which is modified concerning a more realistic beam modelling, avoiding the so-called weak attenuation expansion and considering fluorescence excitation throughout the light cone of the excitation beam. The applicability of the method is demonstrated for synthetic examples as well as microscopic images of chromatographic beads. It is shown that the new method can be successfully applied for reconstructing the true fluorophore distribution in specimens even if the microscopic images are affected by strong attenuation. LAY DESCRIPTION: The applicability of confocal laser scanning microscopy is limited by attenuation of the excitation and the fluorescence emission beam. As a prerequisite for further processing and analysis of the obtained microscopic images, a new method is presented for correcting this attenuation. The correction is based on modeling the excitation as well as the emission beam and on a modified Beer-Lambert law for beam attenuation. The applicability of the method is demonstrated for synthetic examples as well as microscopic images of chromatographic beads. It is shown that the new method can be successfully applied for reconstructing the true fluorophore distribution in specimens even if the microscopic images are affected by strong attenuation.

11.
Aging Clin Exp Res ; 31(6): 863-874, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30806907

RESUMO

BACKGROUND: Aging is associated with declines in muscle mass, strength and quality, leading to physical impairments. An even protein distribution in daily meals has recently been proposed along with adequate total protein intake as important modulators of muscle mass. In addition, due to its short duration, high-intensity interval training (HIIT) has been highlighted as a promising intervention to prevent physical deterioration. However, the interaction between daily protein intake distribution and HIIT intervention in elderlies remain unknown. OBJECTIVE: To investigate muscle adaptation following HIIT in older adults according to daily protein intake distribution. METHODS: Thirty sedentary obese subjects who completed a 12-week elliptical HIIT program were matched [criteria: age (± 2 years), sex, BMI (± 2 kg/m2)] and divided a posteriori into 2 groups according to the amount of protein ingested at each meal: < 20 g in at least one meal (P20-, n = 15, 66.8 ± 3.7 years) and ≥ 20 g in each meal (P20+, n = 15, 68.1 ± 4.1 years). Body composition, functional capacity, muscle strength, muscle power, physical activity level, and nutritional intakes were measured pre- and post-intervention. A two way repeated ANOVA was used to determine the effect of the intervention (HIIT) and protein distribution (P20- vs P20+, p < 0.05). RESULTS: No difference was observed at baseline between groups. Following the HIIT intervention, we observed a significant decrease in waist and hip circumferences and improvements in functional capacities in both P20- and P20 + group (p < 0.05). However, no protein distribution effect was observed. CONCLUSION: A 12-week HIIT program is achievable and efficient to improve functional capacities as well as body composition in obese older adults. However, consuming at least 20 g of proteins in every meal does not further enhance muscle performance in response to a 12-week HIIT intervention.


Assuntos
Proteínas Alimentares/farmacologia , Exercício Físico/fisiologia , Treinamento Intervalado de Alta Intensidade/métodos , Força Muscular , Obesidade/terapia , Idoso , Composição Corporal/fisiologia , Índice de Massa Corporal , Proteínas Alimentares/administração & dosagem , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Força Muscular/efeitos dos fármacos , Força Muscular/fisiologia
12.
Aging Clin Exp Res ; 31(9): 1271-1281, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-30519977

RESUMO

OBJECTIVE: Animal protein sources are considered to be of higher quality than plant protein sources in terms of stimulating muscle metabolism. Our objective was to investigate whether protein intake from animal and plant sources on a daily and per-meal basis differs between healthy older adults with normal and with low muscle mass. METHODS: In this cross-sectional study including 100 healthy, community-dwelling adults (51 women) aged 75-85 years without functional limitations dietary intake was assessed using 7-day food records. Protein intake was classified by six animal and six plant protein sources. Skeletal muscle index (SMI) was determined based on bioelectrical impedance analysis and categorized into 'normal' or 'low' (men ≤ 8.50, women ≤ 5.75 kg/m2). The absolute animal and plant protein intake and their proportion of total protein intake were compared between these groups using Mann-Whitney U test. RESULTS: Daily protein intake was 0.96 ± 0.27 g/kg body weight (BW), 61 ± 10% hereof were from animal origin with no difference between men and women. SMI was low in 39% of men and 35% of women. No differences in absolute daily animal and plant protein intake between participants with normal vs. low SMI were observed. The proportion of animal protein was not different on neither a daily nor a per-meal basis between those with normal and those with low SMI. Women with low SMI consumed less animal protein (in g) for breakfast (4.8 ± 4.1 g vs. 8.5 ± 6.9 g, p = 0.031) and fewer meals per day with at least 50% animal protein (2.2 ± 0.9 vs. 2.7 ± 1.0, p = 0.046) compared to those with normal SMI. CONCLUSION: On a daily basis, the absolute and relative animal protein intake does not differ between healthy older adults without functional limitations with normal vs. low SMI. However, our results indicate that in women animal protein intake on a per-meal basis might be of relevance for the maintenance of muscle mass.


Assuntos
Proteínas de Carne/administração & dosagem , Músculo Esquelético/fisiologia , Proteínas de Vegetais Comestíveis/administração & dosagem , Idoso , Idoso de 80 Anos ou mais , Índice de Massa Corporal , Estudos Transversais , Registros de Dieta , Impedância Elétrica , Ingestão de Energia/fisiologia , Feminino , Humanos , Masculino , Refeições
13.
Biochim Biophys Acta ; 1857(3): 256-65, 2016 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-26619924

RESUMO

The cyanobacterial thylakoid membrane represents a system that can carry out both oxygenic photosynthesis and respiration simultaneously. The organization, interactions and mobility of components of these two electron transport pathways are indispensable to the biosynthesis of thylakoid membrane modules and the optimization of bioenergetic electron flow in response to environmental changes. These are of fundamental importance to the metabolic robustness and plasticity of cyanobacteria. This review summarizes our current knowledge about the distribution and dynamics of electron transport components in cyanobacterial thylakoid membranes. Global understanding of the principles that govern the dynamic regulation of electron transport pathways in nature will provide a framework for the design and synthetic engineering of new bioenergetic machinery to improve photosynthesis and biofuel production. This article is part of a Special Issue entitled: Organization and dynamics of bioenergetic systems in bacteria, edited by Conrad Mullineaux.


Assuntos
Proteínas de Bactérias/metabolismo , Cianobactérias/enzimologia , Complexo de Proteínas da Cadeia de Transporte de Elétrons/metabolismo , Fosforilação Oxidativa , Fotossíntese/fisiologia , Tilacoides/enzimologia
14.
Int J Sport Nutr Exerc Metab ; 27(2): 105-114, 2017 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-27710150

RESUMO

Dietary protein intake should be optimized in all athletes to ensure proper recovery and enhance the skeletal muscle adaptive response to exercise training. In addition to total protein intake, the use of specific proteincontaining food sources and the distribution of protein throughout the day are relevant for optimizing protein intake in athletes. In the present study, we examined the daily intake and distribution of various proteincontaining food sources in a large cohort of strength, endurance and team-sport athletes. Well-trained male (n=327) and female (n=226) athletes completed multiple web-based 24-hr dietary recalls over a 2-4 wk period. Total energy intake, the contribution of animal- and plant-based proteins to daily protein intake, and protein intake at six eating moments were determined. Daily protein intake averaged 108±33 and 90±24 g in men and women, respectively, which corresponded to relative intakes of 1.5±0.4 and 1.4±0.4 g/kg. Dietary protein intake was correlated with total energy intake in strength (r=0.71, p <.001), endurance (r=0.79, p <.001) and team-sport (r=0.77, p <.001) athletes. Animal and plant-based sources of protein intake was 57% and 43%, respectively. The distribution of protein intake was 19% (19±8 g) at breakfast, 24% (25±13 g) at lunch and 38% (38±15 g) at dinner. Protein intake was below the recommended 20 g for 58% of athletes at breakfast, 36% at lunch and 8% at dinner. In summary, this survey of athletes revealed they habitually consume > 1.2 g protein/kg/d, but the distribution throughout the day may be suboptimal to maximize the skeletal muscle adaptive response to training.


Assuntos
Atletas , Desempenho Atlético , Proteínas Alimentares/administração & dosagem , Exercício Físico , Comportamento Alimentar , Esforço Físico , Fenômenos Fisiológicos da Nutrição Esportiva , Adolescente , Adulto , Criança , Estudos de Coortes , Comportamento Alimentar/etnologia , Feminino , Humanos , Internet , Masculino , Pessoa de Meia-Idade , Proteínas do Leite/administração & dosagem , Proteínas Musculares/administração & dosagem , Força Muscular , Países Baixos , Inquéritos Nutricionais , Resistência Física , Proteínas de Vegetais Comestíveis/administração & dosagem , Fenômenos Fisiológicos da Nutrição Esportiva/etnologia , Adulto Jovem
15.
Drug Dev Ind Pharm ; 43(12): 2032-2042, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28756687

RESUMO

OBJECTIVE: The present study aimed at elucidating the influence of polymorphic stability of lipid excipients on the physicochemical characters of different solid lipid microparticles (SLM), with the focus on the alteration of protein distribution in SLM. METHODS: Labeled lysozyme was incorporated into SLM prepared with different excipients, i.e. trimyristin (TG14), glyceryl distearate (GDS), and glyceryl monostearate (GMS), by water-oil-water (w/o/w) or solid-oil-water (s/o/w) method. The distribution of lysozyme in SLM and the release of the protein from SLM were evaluated by confocal laser scanning microscopy. The storage stability of SLM was characterized by HPLC, differential scanning calorimetry, X-ray powder diffraction, and scanning electron microscopy. RESULTS: Lysozyme was displayed as small scattered domains inside GDS and GMS SLM, whereas it was incorporated in the core of TG14 SLM formulated by the w/o/w method or evenly distributed in TG14 SLM prepared by the s/o/w method. Stability study at 37 °C revealed that only TG14 SLM made by the w/o/w method was able to maintain the lysozyme amount both on the particle surface and released from the SLM. Elevated storage temperature induced polymorphic transition of lipids in GDS and GMS SLM, which was, however, not remarkable for the TG14 SLM. CONCLUSIONS: Lipid excipients and particle preparation methods were found to differently affect the lysozyme distribution in SLM, owning to varied storage stabilities of the lipids. The present study provides updated knowledge for rational development of lipid-based formulations for oral delivery of peptide or protein drugs.


Assuntos
Sistemas de Liberação de Medicamentos/métodos , Excipientes/química , Lipídeos/química , Muramidase/química , Proteínas/química , Triglicerídeos/administração & dosagem , Administração Oral , Varredura Diferencial de Calorimetria , Química Farmacêutica , Microscopia Eletrônica de Varredura , Triglicerídeos/química , Difração de Raios X
16.
Zygote ; 23(6): 863-73, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25315095

RESUMO

This study was aimed at investigating zona pellucida glycoproteins (ZP) ZP2, ZP3 mRNA expression as well as ZP3, ZP4 (ZPB) protein distribution before and after in vitro maturation (IVM) in canine oocytes. The cumulus-oocyte complexes (COCs) were recovered from 27 anoestrous mongrel bitches and matured for 72 h in TCM199 medium. The canine COCs were analysed before and after IVM. Using real-time quantitative polymerase chain reaction (RQ-PCR), both groups of oocytes were analysed for detection of ZP2 and ZP3 mRNA profiles as well as using confocal microscopic analysis for observation of ZP3 and ZP4 protein distribution. In post-IVM canine oocytes an increase in transcript content of ZP2 and ZP3 genes as well as a decrease in ZP3 and ZP4 protein levels were observed when compared with pre-IVM oocytes. Moreover, the ZP4 protein before IVM was significantly distributed in the peripheral area of cytoplasm, whereas after IVM it was localized rather than in the entire cytoplasm. In contrast, the ZP3 protein was found both before and after IVM was distributed in the peripheral area of the cytoplasm. In conclusion, we suggest that the expression of ZP2 and ZP3 genes is associated with the maturation stage of canine oocytes, as higher mRNAs levels were found after IVM. However, a decreased expression of ZP3 and ZP4 proteins after IVM suggests maturation-dependent down-regulation of these protein translations, which may result in disturbed fertilization.


Assuntos
Proteínas do Ovo/genética , Proteínas do Ovo/metabolismo , Técnicas de Maturação in Vitro de Oócitos/métodos , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/metabolismo , Oócitos/fisiologia , Receptores de Superfície Celular/genética , Receptores de Superfície Celular/metabolismo , Animais , Células do Cúmulo/citologia , Células do Cúmulo/fisiologia , Cães , Feminino , Regulação da Expressão Gênica , Microscopia Confocal/métodos , Glicoproteínas da Zona Pelúcida
17.
Geriatr Gerontol Int ; 24(6): 634-640, 2024 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-38679586

RESUMO

AIM: Protein intake is an important component in retaining muscle mass, especially among older people. This study examined the relationship between total protein intake and/or the distribution of protein intake in each meal and appendicular muscle mass, using data from the National Health and Nutrition Survey (NHNS) in Japan. METHODS: Data from the NHNS were re-analyzed in this study. We used data from a one-day dietary record, physical examination, and lifestyle questionnaire completed by 1766 participants aged over 60 years. Appendicular muscle mass was assessed by multiple-frequency bioimpedance measurement. Leucine intake was calculated by the weighted average amino acid content from all 98 food subcategories used in the NHNS, based on amino acid composition data in the 2020 Standard Tables of Food Composition. RESULTS: Participants with higher protein intake showed significantly higher appendicular muscle mass. This relationship was independent of physical activity, including steps taken per day, exercise habit, and physical labor. Frequent intake of ≥0.4 g protein/kg/meal was not related to appendicular muscle mass. The combination of higher total protein intake and higher physical activity seemed to have the largest association with appendicular muscle mass. CONCLUSIONS: Higher protein intake may be related to higher appendicular muscle mass, independent of higher physical activity, among older Japanese people. Geriatr Gerontol Int 2024; 24: 634-640.


Assuntos
Proteínas Alimentares , Músculo Esquelético , Inquéritos Nutricionais , Humanos , Japão , Masculino , Feminino , Idoso , Estudos Transversais , Proteínas Alimentares/administração & dosagem , Pessoa de Meia-Idade , Exercício Físico/fisiologia , Idoso de 80 Anos ou mais , Sarcopenia/epidemiologia , Composição Corporal , População do Leste Asiático
18.
Front Cell Dev Biol ; 12: 1420161, 2024.
Artigo em Inglês | MEDLINE | ID: mdl-39376633

RESUMO

A common problem in confocal microscopy is the decrease in intensity of excitation light and emission signal from fluorophores as they travel through 3D specimens, resulting in decreased signal detected as a function of depth. Here, we report a visualization program compatible with widely used fluorophores in cell biology to facilitate image interpretation of differential protein disposition in 3D specimens. Glioblastoma cell clusters were fluorescently labeled for mitochondrial complex I (COXI), P2X7 receptor (P2X7R), ß-Actin, Ki-67, and DAPI. Each cell cluster was imaged using a laser scanning confocal microscope. We observed up to ∼70% loss in fluorescence signal across the depth in Z-stacks. This progressive underrepresentation of fluorescence intensity as the focal plane deepens hinders an accurate representation of signal location within a 3D structure. To address these challenges, we developed ProDiVis: a program that adjusts apparent fluorescent signals by normalizing one fluorescent signal to a reference signal at each focal plane. ProDiVis serves as a free and accessible, unbiased visualization tool to use in conjunction with fluorescence microscopy images and imaging software.

19.
Int J Biol Macromol ; 242(Pt 4): 124909, 2023 Jul 01.
Artigo em Inglês | MEDLINE | ID: mdl-37230453

RESUMO

This study aimed to compare the effect of various exogenous proteins on wheat starch (WS) digestion and assess the relevant mechanisms based on the distribution behaviors of exogenous proteins in the starch matrix. Rice protein (RP), soy protein isolate (SPI), and whey protein isolate (WPI) all effectively suppressed the rapid digestion of WS but with different modes. RP increased the slowly digestible starch content, while SPI and WPI increased the resistant starch content. Fluorescence images showed that RP aggregated and competed for effective space with starch granules, while SPI and WPI formed continuous network structures among the starch matrix. These distribution behaviors endowed different reductions in starch digestion by influencing the gelatinization and ordered structure of starch. Pasting and water mobility results suggested all exogenous proteins inhibited the water migration and swelling of starch. Simultaneously, X-ray diffraction and Fourier transform infrared spectroscopy analysis showed that exogenous proteins improved the ordered structures of starch. RP had a more significant effect on the long-term ordered structure, while SPI and WPI had a more effective effect on the short-term ordered structure. These findings will enrich the theory of exogenous protein inhibiting starch digestion and inspire the applications in low-glycemic index food.


Assuntos
Oryza , Amido , Amido/química , Triticum/química , Proteínas de Soja , Digestão , Água , Oryza/química
20.
J Nutr Sci ; 12: e94, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37649694

RESUMO

Protein intake, sources and distribution impact on muscle protein synthesis and muscle mass in older adults. However, it is less clear whether dietary protein influences muscle strength. Data were obtained from the Researching Eating Activity and Cognitive Health (REACH) study, a cross-sectional study aimed at investigating dietary patterns, cognitive function and metabolic syndrome in older adults aged 65-74 years. Dietary intake was assessed using a 4-d food record and muscle strength using a handgrip strength dynamometer. After adjusting for confounders, in female older adults (n 212), total protein intake (ß = 0⋅22, P < 0⋅01); protein from dairy and eggs (ß = 0⋅21, P = 0⋅03) and plant food sources (ß = 0⋅60, P < 0⋅01); and frequently consuming at least 0⋅4 g/kg BW per meal (ß = 0⋅08, P < 0⋅01) were associated with higher BMI-adjusted muscle strength. However, protein from meat and fish intake and the coefficient of variance of protein intake were not related to BMI-muscle strength in female older adults. No statistically significant associations were observed in male participants (n = 113). There may be sex differences when investigating associations between protein intake and muscle strength in older adults. Further research is needed to investigate these sex differences.


Assuntos
Força da Mão , Vida Independente , Feminino , Masculino , Animais , Nova Zelândia , Estudos Transversais , Força Muscular , Ovos
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