Golgi/plastid‐type manganese superoxide dismutase involved in heat‐stress tolerance during grain filling of rice

Superoxide dismutase (SOD) is widely assumed to play a role in the detoxification of reactive oxygen species caused by environmental stresses. We found a characteristic expression of manganese SOD 1 (MSD1) in a heat‐stress‐tolerant cultivar of rice (Oryza sativa). The deduced amino acid sequence contains a signal sequence and an N‐glycosylation site. Confocal imaging analysis of rice and onion cells transiently expressing MSD1‐YFP showed MSD1‐YFP in the Golgi apparatus and plastids, indicating that MSD1 is a unique Golgi/plastid‐type SOD. To evaluate the involvement of MSD1 in heat‐stress tolerance, we generated transgenic rice plants with either constitutive high expression or suppression of MSD1. The grain quality of rice with constitutive high expression of MSD1 grown at 33/28 °C, 12/12 h, was significantly better than that of the wild type. In contrast, MSD1‐knock‐down rice was markedly susceptible to heat stress. Quantitative shotgun proteomic analysis indicated that the overexpression of MSD1 up‐regulated reactive oxygen scavenging, chaperone and quality control systems in rice grains under heat stress. We propose that the Golgi/plastid MSD1 plays an important role in adaptation to heat stress.     

In Situ Analysis of Solvent and Additive Effects on Film Morphology Evolution in Spin‐Cast Small‐Molecule and Polymer Photovoltaic Materials

To elucidate the details of film morphology/order evolution during spin‐coating, solvent and additive effects are systematically investigated for three representative organic solar cell (OSC) active layer materials using combined in situ grazing incidence wide angle x‐ray scattering (GIWAXS) and optical reflectance. Two archetypical semiconducting donor (p‐type) polymers, P3HT and PTB7, and semiconducting donor small‐molecule, p‐DTS(FBTTh₂)₂ are studied using three neat solvents (chloroform, chlorobenzene, 1,2‐dichlorobenzene) and four processing additives (1‐chloronaphthalene, diphenyl ether, 1,8‐diiodooctane, and 1,6‐diiodohexane). In situ GIWAXS identifies several trends: 1) for neat solvents, rapid crystallization occurs that risks kinetically locking the material into multiple crystal structures or crystalline orientations; and 2) for solvent + additive processed films, morphology evolution involves sequential transformations on timescales ranging from seconds to hours, with key divergences dependent on additive/semiconductor molecular interactions. When π‐planes dominate the additive/semiconductor interactions, both polymers and small molecule films follow similar evolutions, completing in 1–5 min. When side chains dominate the additive/semiconductor interactions, polymer film maturation times are up to 9 h, while initial crystallization times <10 s are observed for small‐molecule films. This study offers guiding information on OSC donor intermediate morphologies, evolution timescales, and divergent evolutions that can inform OSC manufacture.     

Intracellular Transport Dynamics of Endosomes Containing DNA Polyplexes along the Microtubule Network

We have explored the transport of DNA polyplexes enclosed in endosomes within the cellular environment by multiple particle tracking (MPT). The polyplex-loaded endosomes demonstrate enhanced diffusion at short timescales (t < 7 s) with their mean-square displacement (MSD) 〈Δx(t)²〉 scaling as t^1.25. For longer time intervals they exhibit subdiffusive transport and have an MSD scaling as t^0.7. This crossover from an enhanced diffusion to a subdiffusive regime can be explained by considering the action of motor proteins that actively transport these endosomes along the cellular microtubule network and the thermal bending modes of the microtubule network itself.     

Visualization of the post‐Golgi vesicle‐mediated transportation of TGF‐β receptor II by quasi‐TIRFM

Transforming growth factor β receptor II (Tβ RII) is synthesized in the cytoplasm and then transported to the plasma membrane of cells to fulfil its signalling duty. Here, we applied live‐cell fluorescence imaging techniques, in particular quasi‐total internal reflection fluorescence microscopy, to imaging fluorescent protein‐tagged Tβ RII and monitoring its secretion process. We observed punctuate‐like Tβ RII‐containing post‐Golgi vesicles formed in MCF7 cells. Single‐particle tracking showed that these vesicles travelled along the microtubules at an average speed of 0.51 μm/s. When stimulated by TGF‐β ligand, these receptor‐containing vesicles intended to move towards the plasma membrane. We also identified several factors that could inhibit the formation of such post‐Golgi vesicles. Although the inhibitory mechanisms still remain unknown, the observed characteristics of Tβ RII‐containing vesicles provide new information on intracellular Tβ RII transportation. It also renders Tβ RII a good model system for studying post‐Golgi vesicle‐trafficking and protein transportation. (© 2014 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)     

Optimizing Water Exchange Rates and Rotational Mobility for High‐Relaxivity of a Novel Gd‐DO3A Derivative Complex Conjugated to Inulin as Macromolecular Contrast Agents for MRI

Thanks to the understanding of the relationships between the residence lifetime τ_M of the coordinated water molecules to macrocyclic Gd‐complexes and the rotational mobility τ_R of these structures, and according to the theory for paramagnetic relaxation, it is now possible to design macromolecular contrast agents with enhanced relaxivities by optimizing these two parameters through ligand structural modification. We succeeded in accelerating the water exchange rate by inducing steric compression around the water binding site, and by removing the amide function from the DOTA‐AA ligand [1,4,7,10‐tetraazacyclododecane‐1,4,7,10‐tetraacetic acid mono(p‐aminoanilide)] ( L ) previously designed. This new ligand 10[2(1‐oxo‐1‐p‐propylthioureidophenylpropyl]‐1,4,7,10‐tetraazacyclodecane‐1,4,7‐tetraacetic acid ( L ₁ ) was then covalently conjugated to API [O‐(aminopropyl)inulin] to get the complex API ‐(GdL ₁ )x with intent to slow down the rotational correlation time (τ_R) of the macromolecular complex. The evaluation of the longitudinal relaxivity at different magnetic fields and the study of the ¹⁷O‐NMR at variable temperature of the low‐molecular‐weight compound ( GdL ₁ ) showed a slight decrease of the τ_M value ( = 331 ns vs.  = 450 ns for the Gd L complex). Consequently to the increase of the size of the API ‐(GdL ₁ )x complex, the rotational correlation time becomes about 360 times longer compared to the monomeric GdL ₁ complex (τ_R = 33,700 ps), which results in an enhanced proton relaxivity.     

Fasting Plasma Insulin Modulates Lipid Levels and Particle Sizes in 2‐ to 3‐Year‐Old Children

Objective: Obesity and hyperinsulinemia are associated with dyslipidemia in adults and older children, but little is known about these relationships in very young children. We examined the relation of fasting insulin to lipid levels and lipid particle size in young healthy children. Research Methods and Procedures: Analyses were performed on data from 491 healthy 2‐ and 3‐year old Hispanic children enrolled in a dietary study conducted in New York City, 1992–1995. Obesity measures included BMI, ponderal index, skinfold thickness, and waist circumference. Low‐density lipoprotein (LDL)‐ and high‐density lipoprotein (HDL)‐cholesterol particle size were measured by nuclear magnetic resonance. Results: Fasting insulin level was positively correlated with triglyceride levels (r = 0.24 for boys and r = 0.23 for girls; p < 0.001 for both) and inversely correlated with HDL‐cholesterol level in boys (r = ?0.20; p < 0.01). Higher fasting insulin level was also correlated with smaller mean HDL particle size in both boys (r = ?0.21; p < 0.001) and girls (r = ?0.14; p < 0.05) and smaller mean LDL particle size in boys (r = ?0.13; p < 0.05). The associations of fasting insulin level with triglyceride and HDL‐cholesterol levels and HDL and LDL particle size remained significant after multivariate regression adjustment for age, sex, and BMI or ponderal index. Discussion: Fasting insulin level is associated with relative dyslipidemia in healthy 2‐ and 3‐year‐old Hispanic children.     

A work‐for‐food enrichment program increases exploration and decreases stereotypies in four species of bears

Zoo‐housed bears are prone to exhibiting stereotypic behaviors, generally considered indicators of negative welfare. We explored the effects of a variable‐time feeding enrichment schedule on behavioral indicators of welfare in four bear species at Cleveland Metroparks Zoo. We distributed the diets of eight bears in one of five enrichment items, for two consecutive days each, and monitored behavior throughout the day. In Experiment 1, we compared variable‐time to fixed‐time presentation of enrichment over two, 10‐day periods. Overall, bears performed more exploratory behavior when enriched (p < 0.0001). Furthermore, variable‐time enrichment was associated with a greater increase in exploratory behavior than fixed‐time enrichment when compared to baseline (p < 0.001). Both fixed‐time (p_unadjusted <0.05, p_adjusted = 0.07) and variable‐schedule (p_unadjusted <0.05, p_adjusted = 0.09) enrichment were also associated with similar decreases in abnormal behavior compared to baseline. For Experiment 2, we tested habituation to enrichment over 30 days using multiple items and a semi‐variable presentation schedule. Again during the enrichment period, bears exhibited increased exploratory behavior (p < 0.0001) and decreased abnormal behaviors compared to baseline (p_unadjusted = 0.05, p_adjusted = 0.09). We observed no habituation during the 30‐day sustained enrichment period for these behaviors. Collectively, these results suggest that daily, variable‐schedule feeding enrichment, with intermittent presentation of unique enrichment items, increases behavioral indicators of positive welfare and decreases behavioral indicators of negative welfare.     

NMDA receptors on zebrafish Mauthner cells require CaMKII‐α for normal development

Calcium/calmodulin dependent protein kinase 2 (CaMKII) is a multifunctional protein that is highly enriched in the synapse. It plays important roles in neuronal functions such as synaptic plasticity, synaptogenesis, and neural development. Gene duplication in zebrafish has resulted in the occurrence of seven CaMKII genes (camk2a, camk2b1, camk2b2, camk2g1, camk2g2, camk2d1, and camk2d2) that are developmentally expressed. In this study, we used single cell, real‐time quantitative PCR to investigate the expression of CaMKII genes in individual Mauthner cells (M‐cells) of 2 days post fertilization (dpf) zebrafish embryos. We found that out of seven different CaMKII genes, only the mRNA for CaMKII‐α was expressed in the M‐cell at detectable levels, while all other isoforms were undetectable. Morpholino knockdown of CaMKII‐α had no significant effect on AMPA synaptic currents (mEPSCs) but decreased the amplitude of NMDA mEPSCs. NMDA events exhibited a biexponential decay with τ_fast ≈ 30 ms and τ_slow ≈ 300 ms. Knockdown of CaMKII‐α specifically reduced the amplitude of the slow component of the NMDA‐mediated currents (mEPSCs), without affecting the fast component, the frequency, or the kinetics of the mEPSCs. Immunolabelling of the M‐cell showed increased dendritic arborizations in the morphants compared with controls, and knockdown of CaMKII‐α altered locomotor behaviors of touch responses. These results suggest that CaMKII‐α is present in embryonic M‐cells and that it plays a role in the normal development of excitatory synapses. Our findings pave the way for determining the function of specific CaMKII isoforms during the early stages of M‐cell development. © 2014 Wiley Periodicals, Inc. Develop Neurobiol 75: 145–162, 2015     

Objectively Measured Physical Activity in a Representative Sample of 3‐ to 4‐Year‐Old Children

Objective: This study aimed to describe levels of physical activity in a representative sample of preschool children and to quantify tracking of activity over 1 year. Research Methods and Procedures: Physical activity (mean accelerometry counts/minute) was assessed over 3 days using the Computer Science and Applications accelerometer in 3‐ to 4‐year‐old children (n = 104; 52 boys; mean age, 3.7 ± 0.4 years). In 60 children (30 boys), measurements were repeated 1 year later. Results: Mean total activity at baseline was 777 ± 207 counts/minute in boys and 657 ± 172 counts/minute for girls; this gender difference was significant (p < 0.001). In the cross‐sectional analysis, total activity was significantly positively related to age (r = 0.37, p = 0.007). In the sample followed up for 1 year, mean total activity was 849 ± 252. The longitudinal analysis confirmed that total physical activity increased over the 1‐year period (paired Student's t test, p < 0.001). The tracking rank order correlation coefficient of total activity count over 1 year was r = 0.40 (p < 0.001). Discussion: This study suggests that total activity increases during the preschool period in Scottish children and that gender differences in total activity are present early in life. Tracking of total activity was only modest, but adequate assessment of tracking requires methodological research aimed at elucidating the biological meaning of accelerometer output.     

Priority effects among young‐of‐the‐year fish: reduced growth of bluegill sunfish (Lepomis macrochirus) caused by yellow perch (Perca flavescens)?

1. When available, Daphnia spp. are often preferred by age‐0 yellow perch and bluegill sunfish because of energetic profitability. We hypothesised that predation by age‐0 yellow perch could lead to a midsummer decline (MSD) of Daphnia spp. and that priority effects may favour yellow perch because they hatch before bluegill, allowing them to capitalise on Daphnia spp. prior to bluegill emergence. 2. Data were collected from 2004 to 2010 in Pelican Lake, Nebraska, U.S.A. The lake experienced a prolonged MSD in all but 1 year (2005), generally occurring within the first 2 weeks of June except in 2008 and 2010 when it occurred at the end of June. MSD timing is not solely related to seasonal patterns of age‐0 yellow perch consumption. Nevertheless, when Daphnia spp. biomass was low during 2004 and 2006–2010 (<4 mg wet weight L^?1), predation by age‐0 yellow perch seems to have suppressed Daphnia spp. biomass (i.e. <1.0 mg wet weight L^?1). The exception was 2005 when age‐0 yellow perch were absent. 3. Growth of age‐0 bluegill was significantly faster in 2005, when Daphnia spp. were available in greater densities (>4 mg wet weight L^?1) compared with the other years (<0.2 mg wet weight L^?1). 4. We conclude that age‐0 yellow perch are capable of reducing Daphnia biomass prior to the arrival of age‐0 bluegill, ultimately slowing bluegill growth. Thus, priority effects favour age‐0 yellow perch when competing with age‐0 bluegill for Daphnia. However, these effects may be minimised if there is a shorter time between hatching of the two species, higher Daphnia spp. densities or lower age‐0 yellow perch densities.     

2‐photon excitation fluorescence microscopy enables deeper high‐resolution imaging of voltage and Ca2+ in intact mice,rat, and rabbit hearts

We describe a novel two‐photon (2P) laser scanning microscopy (2PLSM) protocol that provides ratiometric transmural measurements of membrane voltage (V_m) via Di‐4‐ANEPPS in intact mouse, rat and rabbit hearts with subcellular resolution. The same cells were then imaged with Fura‐2/AM for intracellular Ca²⁺ recordings. Action potentials (APs) were accurately characterized by 2PLSM vs. microelectrodes, albeit fast events (<1 ms) were sub‐optimally acquired by 2PLSM due to limited sampling frequencies (2.6 kHz). The slower Ca²⁺ transient (CaT) time course (>1ms) could be accurately described by 2PLSM. In conclusion, V_m ‐ and Ca²⁺‐sensitive dyes can be 2P excited within the cardiac muscle wall to provide AP and Ca²⁺ signals to ～400 µm. (© 2013 WILEY‐VCH Verlag GmbH &Co. KGaA, Weinheim)     

The influence of RAMP1 overexpression on CGRP‐induced osteogenic differentiation in MG‐63 cells in vitro: An experimental study

The aim of this study was to elucidate the influence of receptor activity modifying protein 1 (RAMP1) overexpression on the expression and distribution of calcitonin receptor‐like receptor (CRLR) in MG‐63 cells. Our research also focused on whether RAMP1 overexpression enhanced the promoting effect of exogenous CGRP on osteogenic differentiation in MG‐63 cells. We first constructed a eukaryotic expression vector containing human RAMP1 and stably transfected it into MG‐63 cells. Real‐time PCR and Western blotting were used to determine the expression levels of RAMP1 and CRLR mRNA and protein, respectively. Immunofluorescence analysis was employed to compare the distribution of CRLR in transfected cells. After treatment with CGRP, the extent of osteogenic differentiation was evaluated by simultaneous monitoring of alkaline phosphatase activity, the expression patterns of osteoblastic markers and mineralisation staining. We found that RAMP1 was more highly expressed in the transfected group compared with the control groups (P < 0.01). The CRLR expression was significantly higher than that in the control groups (P < 0.05). In addition, after 7 days of CGRP treatment to induce osteogenic differentiation, the expression of collagen I mRNA was markedly increased in the transfected group (P < 0.05). The transfected group exhibited more granular precipitation in the cytoplasm with alkaline phosphatase staining after 7 and 14 days of differentiation. When stained with Alizarin Red, cells overexpressing RAMP1 were darker and formed many mineralised nodules with clear boundaries and calcium deposition typical of mineralised bone matrix structures at 28 days post‐induction of differentiation. The CGRP‐induced ALP activity in the RAMP1 overexpression group was significantly higher 3, 6 and 9 days after induction than that in the two control groups (P < 0.05). RAMP1 overexpression promotes CRLR expression, localisation on the cell membrane and enhanced CGRP‐mediated differentiation of MG‐63 cells. This study contributes to a better understanding of the molecular mechanisms governing CGRP‐induced MG‐63 differentiation. J. Cell. Biochem. 114: 314–322, 2013. © 2012 Wiley Periodicals, Inc.     

Visual tracking from a rigid‐hulled inflatable boat to determine foraging movements of breeding terns

ABSTRACT Defining the at‐sea foraging movements of seabirds is fundamental to understanding their ecology and can also be important in assessing the potential impact of marine developments such as offshore wind farms (OWFs). Surveys of predefined areas using aerial or boat‐based transect surveys may not allow adequate assessment of the relative importance of different areas to birds. Individual‐based satellite or radio‐telemetry can be effective in identifying foraging ranges and preferred areas, but may not be suitable for some species. We developed a method to determine the foraging movements of breeding terns (Sterna spp.) by visually tracking individuals using a rigid‐hulled inflatable boat (RHIB). Sandwich Terns (S. sandvicensis), Common Terns (S. hirundo), and Arctic Terns (S. paradisaea) were tracked from colonies in Norfolk and Anglesey, United Kingdom, from 2006 to 2008. The proportion of complete (from and to colony) trips varied from 29–60% among species, years, and colonies. Individual Sandwich Terns were tracked for periods up to 126 min over distances up to 72 km and as far as 54 km from the breeding colony, further than Arctic (up to 57 km and 29 km from the colony) and Common (to 29 km and <9 km from the colony) terns. Mean values were much smaller. Multivariate modeling of Sandwich Tern foraging trips indicated that flight speeds >50 km/hr coupled with greater distances from shore (>25 km) significantly reduced the likelihood of tracking a bird for an entire foraging trip. Use of different boats that differ in speed and performance may alleviate such issues. Visual tracking allowed us to collect data on foraging behavior, flight height, and prey capture rates, and also permitted comparisons between species. Our results indicate that visual tracking may be an effective means of determining the foraging movements and at‐sea behavior of a variety of short‐ranging, day‐active seabirds.     

Rutin treats myocardial damage caused by pirarubicin via regulating miR‐22‐5p‐regulated RAP1/ERK signaling pathway

Our experiments have previously demonstrated that rutin (RUT) can improve myocardial damage caused by pirarubicin (THP). However, the underlying molecular mechanisms remain uncertain. In this study, we developed an microRNA (miRNA) chip by replicating the rat model of THP‐induced myocardial injury and identified miR‐22‐5p and the RAP1‐member of RAS oncogene family/extracellular regulated protein kinases (RAP1/ERK) signaling pathway as an object of study. Also, in vivo experiments demonstrated that THP caused abnormal changes in the electrocardiogram, cardiac function, and histomorphology in rats (P < .01). THP also reduces the expression of miR‐22‐5p (P < .01) and increases the levels of RAP1/ERK signaling pathway‐related proteins (P < .01, P < .05). RUT significantly improved THP‐induced myocardial damage (P < .01), increased the expression of miR‐22‐5p (P < .01), and decreased the levels of RAP1/ERK signaling pathway‐related proteins (P < .01, P < .05). In vitro studies confirmed that Rap1a is one of the target genes of miR‐22‐5p. miR‐22‐5p overexpression in cardiomyocytes can affect the RAP1/ERK pathway and reduce reactive oxygen species production and cardiomyocyte apoptosis caused by THP (P < .01), which is consistent with the effect of RUT. Our results indicate that RUT treats THP‐induced myocardial damage, which may be achieved by upregulating miR‐22‐5p, causing changes in its target gene Rap1a and the RAP1/ERK pathway.     

A mathematical analysis for the Brownian dynamics of a DNA tether

In the single-particle tracking experiment, the internal motion of a single DNA or polymer molecule whose one end is attached to a microsphere (optical marker) and the other end is anchored to a substratum is studied (Finzi and Gelles, 1995). The stochastic Brownian dynamics of the sphere reflect the spontaneous fluctuations, thus the physical characteristics, of the DNA or polymer molecule (Qian and Elson, 1999, Qian, 2000). In this paper, two continuous models of polymer molecules, a flexible elastic string and a weakly bentable elastic rod, are analyzed. Both models are cast mathematically in terms of linear stochastic differential equations. Based on Fourier analyses, we calculate the mean square displacement (MSD) of the particle motion, the key observable in the experiment. We obtain for both models the short-time asymptotics for the MSD, as well as the long-time behavior in terms of the smallest non-zero eigenvalues. It is shown that: (i) the long-time dynamics of continuous elastic string model quantitatively agree with that of the discrete bead-spring model. (ii) The short-time MSD of both models are controlled by the tethered particle, with linear dependence on t. (iii) The two models show characteristic difference for long-time behavior: The longest relaxation time is proportional to L ² for long elastic string and to L for short elastic string, but is proportional to L ⁴ for both long and short weakly bentable rod. Received: 26 March 1998 / Revised version: 9 June 2000 / Published online: 14 September 2000     

Synthesis and application of Nα‐Fmoc‐Nπ‐4‐methoxybenzyloxymethylhistidine in solid phase peptide synthesis

The 4‐methoxybenzyloxymethyl (MBom) group was introduced at the N^π‐position of the histidine (His) residue by using a regioselective procedure, and its utility was examined under standard conditions used for the conventional and the microwave (MW)‐assisted solid phase peptide synthesis (SPPS) with 9‐fluorenylmethyoxycarbonyl (Fmoc) chemistry. The N^π‐MBom group fulfilling the requirements for the Fmoc strategy was found to prevent side‐chain‐induced racemization during incorporation of the His residue even in the case of MW‐assisted SPPS performed at a high temperature. In particular, the MBom group proved to be a suitable protecting group for the convergent synthesis because it remains attached to the imidazole ring during detachment of the protected His‐containing peptide segments from acid‐sensitive linkers by treatment with a weak acid such as 1% trifluoroacetic acid in dichloromethane. We also demonstrated the facile synthesis of Fmoc‐His(π‐MBom)‐OH with the aid of purification procedure by crystallization to effectively remove the undesired τ‐isomer without resorting to silica gel column chromatography. This means that the present synthetic procedure can be used for large‐scale production without any obstacles. Copyright © 2012 European Peptide Society and John Wiley & Sons, Ltd.     

The non‐coding RNA OTUB1‐isoform2 promotes ovarian tumour progression and predicts poor prognosis

Ovarian cancer is the leading malignancy of the female reproductive system and is associated with inconspicuous early invasion and metastasis. We have previously reported that the oncogene OTUB1 plays a crucial role in ovarian cancer progression, but the role of its isoform, the non‐coding RNA OTUB1‐isoform2, in ovarian cancer is still elusive. Here, we reported that OTUB1‐isoform2 expression in ovarian cancer tissues was significantly higher than that in the paired paratumorous tissues (P < .01). The patients with high expression of OTUB1‐isoform2 had larger tumours than those with low expression (P < .05). The high expression of OTUB1‐isoform2 was correlated with the involvement of bilateral ovaries (P < .05), lymph node metastasis (P < .05), vascular invasion (P < .05), greater omentum involvement (P < .01), fallopian tube involvement (P < .05), advanced FIGO stages (P < .01) and recurrence (P < .01). Moreover, OTUB1‐isoform2 served as an independent negative prognostic predictor for disease‐free survival (DFS) and disease‐specific survival (DSS). Overexpression of OTUB1‐isoform2 in the ovarian cancer cells stimulated cell proliferation, migration and invasion both in vitro and in vivo. In summary, our study suggested that OTUB1‐isoform2 is a novel prognostic biomarker with independent oncogenic functions for ovarian cancer.     

Effects of a School‐based Weight Maintenance Program for Mexican‐American Children: Results at 2 Years

The prevalence of childhood overweight has increased significantly, with the highest rates noted among Mexican Americans. Many negative health outcomes are associated with overweight; thus, there is a need for effective weight‐loss interventions tailored to this group. This study evaluated 24‐month outcomes of a randomized, controlled trial involving an intensive lifestyle‐based weight maintenance program targeting overweight Mexican‐American children at a charter school in Houston, Texas. A total of 60 children (33 males, 55%) between the ages of 10 and 14 at or >85th percentile for BMI were recruited. Participants were randomized to an instructor‐led intervention (ILI) or a self‐help (SH) program, both aimed at modifying eating and physical activity behaviors using behavior modification strategies. Changes in participants' standardized BMI (zBMI) were assessed at baseline, 1, and 2 years. Tricep skinfold, total cholesterol, triglycerides, high‐density lipoprotein cholesterol, and calculated low‐density lipoprotein were assessed at baseline and 1 year. ILI participants showed significantly greater decreases in zBMI at 1 and 2 years (F = 26.8, P < 0.001, F = 4.1, P < 0.05, respectively) compared to SH controls. ILI participants showed greater improvements in body composition, as measured by tricep skinfold (F = 9.75, P < 0.01). Children in the ILI condition experienced benefits with respect to total cholesterol (F = 7.19, P < 0.05) and triglycerides (F = 4.35, P < 0.05) compared to children in the SH condition. Overall, the school‐based intervention resulted in improved weight and clinical outcomes in overweight Mexican‐American children, and zBMI was maintained over 2 years.