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1.
Crit Rev Food Sci Nutr ; : 1-12, 2023 Oct 09.
Article in English | MEDLINE | ID: mdl-37811643

ABSTRACT

Health effects of vegan diets among children and adolescents are a controversial public health topic. Thus, the aim of the present systematic review is to evaluate a broad range of health outcomes among vegan children and adolescents aged 0 to 18 years. 18 studies met the inclusion criteria (17 cross-sectional, 1 RCT). Meta-analyses showed lower protein, calcium, vitamin B2, saturated fatty acid, and cholesterol intakes, and lower ferritin, HDL and LDL levels as well as height in vegan compared to omnivorous children/adolescents. Higher intakes of carbohydrates, polyunsaturated fatty acids, fiber, folate, vitamins C and E, magnesium, iron, and potassium were observed in vegans. Blood levels of vitamin B12 were higher among vegan children due to supplement use. Single study results suggested further differences between vegan and non-vegan children, such as lower bone mineral content or urinary iodine among vegan children. Risk of Bias was rated as high or very high in 7 out of 18 studies. The certainty of evidence for the meta-analyses was low (n = 2) or very low (n = 46). Overall, the available evidence points to both risks and benefits associated with a vegan diet among children, although more and better designed studies are needed.

2.
Sci Rep ; 13(1): 7848, 2023 05 15.
Article in English | MEDLINE | ID: mdl-37188711

ABSTRACT

Practicing complex everyday life walking activities is challenging in paediatric neurorehabilitation, although it would prepare patients more comprehensively for the requirements of daily life. Floor projections allow simulation and training of such situations in therapy. Twenty healthy youths aged 6-18 years stepped over a tree trunk and balanced over kerbstones in a real and projected condition. Spatiotemporal and kinematic parameters of the two conditions were compared by equivalence analysis, using the medians of the differences between the two conditions with their bootstrapped 95% confidence intervals. Velocity, step and stride length, step width, and single support time were generally equivalent between the two conditions. Knee and hip joint angles and toe clearance decreased substantially during the execution phase of the projected tree trunk condition. The largest differences were found at the end of the execution phase in both tasks for the ankle joints. As spatiotemporal parameters were equivalent between the conditions, floor projections seem suitable to train precise foot placement. However, differences in knee and hip joint kinematics and toe clearance revealed that floor projections are not applicable for obstacles with a vertical extension. Therefore, exercises aiming at knee and hip flexion improvement should favourably be trained with real objects.


Subject(s)
Gait , Walking , Humans , Adolescent , Child , Lower Extremity , Foot , Knee Joint , Biomechanical Phenomena
3.
JMIR Rehabil Assist Technol ; 9(4): e39687, 2022 Dec 16.
Article in English | MEDLINE | ID: mdl-36525299

ABSTRACT

BACKGROUND: Objective measures specifically assessing selective voluntary motor control are scarce. Therefore, we have developed an interval-scaled assessment based on accelerometers. OBJECTIVE: This study provided a preliminary evaluation of the validity and reliability of this novel gamelike assessment measuring lower limb selective voluntary motor control in children with cerebral palsy (CP). METHODS: Children with CP and their neurologically intact peers were recruited for this psychometric evaluation of the assessgame. The participants played the assessgame and steered an avatar by selective hip, knee, or ankle joint movements captured with accelerometers. The assessgame's scores provide information about the accuracy of the selective movement of the target joint and the amplitude and frequency of involuntary movements occurring in uninvolved joints. We established discriminative validity by comparing the assessgame scores of the children with CP with those of the neurologically intact children, concurrent validity by correlations with clinical scores and therapists' opinions, and relative and absolute test-retest reliability. RESULTS: We included 20 children with CP (mean age 12 years and 5 months, SD 3 years and 4 months; Gross Motor Function Classification System levels I to IV) and 31 neurologically intact children (mean age 11 years and 1 month, SD 3 years and 6 months). The assessgame could distinguish between the children with CP and neurologically intact children. The correlations between the assessgame's involuntary movement score and the therapist's rating of the occurrence of involuntary movements during the game were moderate (Spearman ρ=0.56; P=.01), whereas the correlations of the assessgame outcomes with the Selective Control Assessment of the Lower Extremity and Gross Motor Function Classification System were low and not significant (|ρ|≤0.39). The intraclass correlation coefficients were >0.85 and indicated good relative test-retest reliability. Minimal detectable changes amounted to 25% (accuracy) and 44% (involuntary movement score) of the mean total scores. The percentage of children able to improve by the minimal detectable change without reaching the maximum score was 100% (17/17) for the accuracy score and 94% (16/17) for the involuntary movement score. CONCLUSIONS: The assessgame proved reliable and showed discriminative validity in this preliminary evaluation. Concurrent validity was moderate with the therapist's opinion but relatively poor with the Selective Control Assessment of the Lower Extremity. We assume that the assessment's gamelike character demanded various other motor control aspects that are less considered in current clinical assessments.

4.
Clin Neurophysiol Pract ; 7: 107-114, 2022.
Article in English | MEDLINE | ID: mdl-35372733

ABSTRACT

Objective: To quantify selective voluntary motor control (SVMC) objectively and more precisely, we combined the "Selective Control Assessment of the Lower Extremity" (SCALE) with surface electromyography. The resulting Similarity Index (SI) measures the similarity of muscle activation patterns. This study evaluated the preliminary validity and reliability of this novel SISCALE measure in children with cerebral palsy (CP). Methods: We investigated concurrent validity by correlating the SISCALE of 24 children with CP (median age 10.6 years) with comparator assessments. For discriminative validity, the patients' SISCALE scores were compared to 31 neurologically intact age-matched peers. Test-retest reliability was quantified using intraclass correlation coefficients (ICC) and minimal detectable change (MDC) values. Results: The SISCALE correlated strongly with the SCALE (ρ = 0.90, p < .001) and the Gross Motor Function Classification System (ρ = -0.74, p < .001). SISCALE scores were significantly lower in children with CP compared to healthy peers. Test-retest reliability appeared good (for the more and less affected leg, ICC ≥ 0.84, and MDC ≤ 0.17). Conclusions: Validity and reliability of the SISCALE leg and total scores lay within clinically acceptable ranges. Further clinimetric analyses should include responsiveness. Significance: A neurophysiology-based assessment could contribute to a more refined assessment of SVMC impairments.

5.
Disabil Rehabil ; 44(14): 3694-3700, 2022 07.
Article in English | MEDLINE | ID: mdl-33577357

ABSTRACT

PURPOSE: We evaluated the validity and intra-, inter-, and test-retest reliability of the Selective Control of the Upper Extremity Scale (SCUES) sum and item scores in patients with upper motor neuron lesions. METHODS: Thirty-one boys and 15 girls (mean age ± SD: 11 years 1 month ± 3 years 9 month) with upper motor neuron lesions participated. We correlated SCUES scores with the range of motion items of the Melbourne Assessment 2 (MA2) and Box and Block Test (BBT) to establish concurrent validity and compared scores between the more and less affected side for discriminative validity. Intra-class correlation coefficients (ICC) and smallest detectable changes (SDC) indicated relative and absolute reliability. RESULTS: For the more affected side, SCUES sum scores correlated well with MA2 (ρ = 0.83) and BBT (ρ = 0.77), and reliability proved high for intra-rater (ICC = 0.93, SDC = 2.55), inter-rater (ICC = 0.86, SDC = 3.58), and test-retest (ICC = 0.98, SDC = 1.41) reliability. Reliability of single items varied from 0.64 (inter-rater elbow) to 0.98 (intra-rater elbow). Limb and item scores were lower for the more affected side. CONCLUSION: The SCUES limb and item scores seem valid and reliable in children with upper motor neuron lesions. While future studies should evaluate the responsiveness of the SCUES, we recommend that the same rater should score a patient twice.Implications for rehabilitationThe SCUES assesses selective voluntary motor control and appears valid and reliable in patients with upper motor neuron lesions.Test-retest reliability of the SCUES seems excellent.SCUES single item scores show concurrent validity and acceptable reliability.Limb and item scores are significantly lower for the more affected side.


Subject(s)
Motor Neurons , Upper Extremity , Child , Female , Humans , Male , Range of Motion, Articular/physiology , Reproducibility of Results
6.
Phys Ther ; 101(8)2021 08 01.
Article in English | MEDLINE | ID: mdl-34464449

ABSTRACT

OBJECTIVE: The purpose of this study was to evaluate the relative importance of different approaches to measure upper extremity selective voluntary motor control (SVMC), spasticity, strength, and trunk control for explaining self-care independence in children affected by upper motor neuron lesions. METHODS: Thirty-one patients (mean [SD] age = 12.5 [3.2] years) with mild to moderate arm function impairments participated in this observational study. Self-care independence was evaluated with the Functional Independence Measure for children (WeeFIM). Upper extremity SVMC was quantified with the Selective Control of the Upper Extremity Scale (SCUES), a similarity index (SISCUES) calculated from simultaneously recorded surface electromyography muscle activity patterns, and an accuracy and involuntary movement score derived from an inertial-measurement-unit-based assessgame. The Trunk Control Measurement Scale was applied and upper extremity spasticity (Modified Ashworth Scale) and strength (dynamometry) were assessed. To determine the relative importance of these factors for self-care independence, 3 regression models were created: 1 included only upper extremity SVMC measures, 1 included upper extremity and trunk SVMC measures (overall SVMC model), and 1 included all measures (final self-care model). RESULTS: In the upper extremity SVMC model (total variance explained 52.5%), the assessgame (30.7%) and SCUES (16.5%) were more important than the SISCUES (4.5%). In the overall SVMC model (75.0%), trunk SVMC (39.0%) was followed by the assessgame (21.1%), SCUES (11.0%), and SISCUES (4.5%). In the final self-care model (82.1%), trunk control explained 43.2%, upper extremity SVMC explained 23.1%, spasticity explained 12.3%, and strength explained 2.3%. CONCLUSION: Although upper extremity SVMC explains a substantial portion of self-care independence, overall trunk control was even more important. Whether training trunk control and SVMC can translate to improved self-care independence should be the subject of future research. IMPACT: This study highlights the importance of trunk control and selective voluntary motor control for self-care independence in children with upper motor neuron lesions.


Subject(s)
Motor Neuron Disease/physiopathology , Muscle Spasticity/physiopathology , Self Care , Torso/physiopathology , Upper Extremity/physiopathology , Adolescent , Child , Electromyography , Female , Humans , Male
7.
JMIR Serious Games ; 9(3): e26028, 2021 Jul 28.
Article in English | MEDLINE | ID: mdl-34319236

ABSTRACT

BACKGROUND: Computer game-based interventions are emerging in pediatric neurorehabilitation, as they can provide two key elements for motor learning-motivating environments that enable long-term compliance, which is particularly relevant for children, and augmented feedback for improving movement performance. OBJECTIVE: The overall aim of this study is to develop an interactive computer play for children with upper motor neuron lesions to train selective voluntary motor control and give particular attention to motivation and feedback. We also aim to determine features that make games engaging, investigate which sensory feedback modality is noticed the fastest during play, develop an interactive game, and evaluate its feasibility. METHODS: We identified engaging game features by interviewing 19 children and adolescents undergoing rehabilitation. By using a test version of the game, we determined the response times of 10 patients who had to react to visual, auditory, or combined feedback signals. On the basis of the results of these two subprojects, we developed and designed a game environment. Feasibility was studied in terms of the practicability and acceptability of the intervention among 5 children with upper motor neuron lesions. RESULTS: The game features deemed the most important by pediatric patients were strategic gameplay (13/29, 45% of answers) and choice (6/29, 21%). While playing the game, an acoustic alarm signal (reaction time: median 2.8 seconds) was detected significantly faster (P=.01) than conditions with other feedback modalities (avatar velocity reduction: median 7.8 seconds; color desaturation: median 5.7 seconds). Most children enjoyed playing the game, despite some technical issues. CONCLUSIONS: The careful identification of game features that increase motivation and feedback modalities that inform children quickly led to the development of an interactive computer play for training selective voluntary motor control in children and adolescents with upper motor neuron lesions.

8.
Hum Mov Sci ; 77: 102790, 2021 Jun.
Article in English | MEDLINE | ID: mdl-33798928

ABSTRACT

BACKGROUND: Neurophysiological development of selective voluntary motor control (SVMC) is assumed but has not been quantified objectively. We assessed SVMC with (i) clinical assessments, (ii) a combination of these assessments with surface electromyography (sEMG) and, (iii) a playful computer game. The aim of this study was to describe and compare age-related differences in SVMC, quantified with these tools, in neurologically intact children, adolescents, and adults. METHODS: We measured upper and lower extremity SVMC with three assessments in 31 children and adolescents. A sample of 33 and 31 adults provided reference values for the upper and lower extremity assessments, respectively. The Selective Control of the Upper Extremity Scale (SCUES) or the Selective Control Assessment of the Lower Extremity (SCALE) were combined with simultaneous sEMG recordings. We quantified SVMC by a similarity index that compared an individual's muscle activation pattern with those of an adult reference group. The SVMC Assessgame required isolated joint movements to steer an avatar and quantified the accuracy of the selective movement and the extent of involuntary movements occurring in not involved joints. RESULTS: Results from the conventional clinical assessments correlated low to moderately with age (SCUES: r = 0.55, p = 0.013; SCALE: r = 0.44, p = 0.001), while the correlation between the sEMG based similarity index and age was negligible (r ≤ 0.25). The outcomes of the Assessgame correlated highly with age (r ≥ 0.80, p ≤ 0.001). Older children and adolescents performed movements more accurately and with fewer involuntary movements compared to younger participants. CONCLUSIONS: The tools assess and quantify SVMC differently, affecting the way they capture age-related differences in SVMC. Some assessments require reference values from neurologically intact children and adolescents to correctly classify impairments of SVMC in patients with neuromotor disorders.


Subject(s)
Electromyography , Lower Extremity/physiology , Motor Skills/physiology , Movement , Upper Extremity/physiology , Adolescent , Adult , Age Factors , Child , Cross-Sectional Studies , Female , Humans , Male , Middle Aged , Reference Values , Reproducibility of Results , Video Games , Young Adult
9.
Sci Prog ; 104(2): 368504211008058, 2021.
Article in English | MEDLINE | ID: mdl-33871293

ABSTRACT

Current clinical assessments evaluating selective voluntary motor control are measured on an ordinal scale. We combined the Selective Control of the Upper Extremity Scale (SCUES) with surface electromyography to develop a more objective and interval-scaled assessment of selective voluntary motor control. The resulting Similarity Index (SI) quantifies the similarity of muscle activation patterns. We aimed to evaluate the validity and reliability of this new assessment named SISCUES (Similarity Index of the SCUES) in children with upper motor neuron lesions. Thirty-three patients (12.2 years [8.8;14.9]) affected by upper motor neuron lesions with mild to moderate impairments and 31 typically developing children (11.6 years [8.5;13.9]) participated. We calculated reference muscle activation patterns for the SISCUES using data of 33 neurologically healthy adults (median [1st; 3rd quantile]: 32.5 [27.9; 38.3]). We calculated Spearman correlations (ρ) between the SISCUES and the SCUES and the Manual Ability Classification System (MACS) to establish concurrent validity. Discriminative validity was tested by comparing scores of patients and healthy peers with a robust ANCOVA. Intraclass correlation coefficients2,1 and minimal detectable changes indicated relative and absolute reliability. The SISCUES correlates strongly with SCUES (ρ = 0.76, p < 0.001) and moderately with the MACS (ρ = -0.58, p < 0.001). The average SISCUES can discriminate between patients and peers. The intraclass correlation coefficient2,1 was 0.90 and the minimal detectable change was 0.07 (8% of patients' median score). Concurrent validity, discriminative validity, and reliability of the SISCUES were established. Further studies are needed to evaluate whether it is responsive enough to detect changes from therapeutic interventions.


Subject(s)
Motor Neuron Disease , Upper Extremity , Adult , Child , Electromyography , Humans , Motor Neuron Disease/diagnosis , Motor Neurons , Reproducibility of Results
10.
Front Neurol ; 11: 572038, 2020.
Article in English | MEDLINE | ID: mdl-33343485

ABSTRACT

Background: Impaired selective voluntary motor control is defined as "the reduced ability to isolate the activation of muscles in response to demands of a voluntary posture or movement." It is a negative motor sign of an upper motor neuron lesion. Objective: This paper reviews interventions that may improve selective motor control in children and youths with spastic cerebral palsy. The aim was to systematically evaluate the methodological quality and formulate the level of evidence from controlled studies. Methods: Six databases (Scopus, Web of Science, PubMed, Embase, MEDLINE, and CINAHL) were searched with predefined search terms for population, interventions, and outcomes. Two reviewers independently completed study selection and ratings of methodological quality and risk of bias. Evidence was summarized in a best evidence synthesis. Results: Twenty-three studies from initially 2,634 papers were included. The interventions showed a wide variety of approaches, such as constraint-induced movement therapy (CIMT), electrical stimulation, robot-assisted therapy, and functional training. The evidence synthesis revealed conflicting evidence for CIMT, robot-assisted rehabilitation and mirror therapy for the upper extremities in children with cerebral palsy. Conclusions: Final recommendations are difficult due to heterogeneity of the reviewed studies. Studies that include both an intervention and an outcome that specifically focus on selective voluntary motor control are needed to determine the most effective therapy.

11.
J Neuroeng Rehabil ; 17(1): 89, 2020 07 13.
Article in English | MEDLINE | ID: mdl-32660569

ABSTRACT

INTRODUCTION: Current clinical assessments measure selective voluntary motor control (SVMC) on an ordinal scale. We introduce a playful, interval-scaled method to assess SVMC in children with brain lesions and evaluate its validity and reliability. METHODS: Thirty-one neurologically intact children (median [1st-3rd quartile]: 11.6 years [8.5-13.9]) and 33 patients (12.2 years [8.8-14.9]) affected by upper motor neuron lesions with mild to moderate impairments participated. Using accelerometers, they played a movement tracking game (assessgame) with isolated joint movements (shoulder, elbow, lower arm [pro-/supination], wrist, and fingers), yielding an accuracy score. Involuntary movements were recorded simultaneously and resulted in an involuntary movement score. Both scores were normalized to the performance of 33 neurologically intact adults (32.5 years [27.9; 38.3]), which represented physiological movement patterns. We correlated the assessgame outcomes with the Manual Ability Classification System, Selective Control of the Upper Extremity Scale, and a therapist rating of involuntary movements. Furthermore, a robust ANCOVA was performed with age as covariate, comparing patients to their healthy peers at the age levels of 7.5, 9, 10.5, 12, and 15 years. Intraclass correlation coefficients and smallest real differences indicated relative and absolute reliability. RESULTS: Correlations (Kendall/Spearman) for the accuracy score were τ = 0.29 (p = 0.035; Manual Ability Classification System), ρ = - 0.37 (p = 0.035; Selective Control of the Upper Extremity Scale), and ρ = 0.64 (p < 0.001; therapist rating). Correlations for the involuntary movement metric were τ = 0.37 (p = 0.008), ρ = - 0.55 (p = 0.001), and ρ = 0.79 (p < 0.001), respectively. The robust ANCOVAs revealed that patients performed significantly poorer than their healthy peers in both outcomes and at all age levels except for the dominant/less affected arm, where the youngest age group did not differ significantly. Robust intraclass correlation coefficients and smallest real differences were 0.80 and 1.02 (46% of median patient score) for the accuracy and 0.92 and 2.55 (58%) for involuntary movements, respectively. CONCLUSION: While this novel assessgame is valid, the reliability might need to be improved. Further studies are needed to determine whether the assessgame is sensitive enough to detect changes in SVMC after a surgical or therapeutic intervention.


Subject(s)
Accelerometry/instrumentation , Accelerometry/methods , Motor Neuron Disease/physiopathology , Psychomotor Performance/physiology , Adolescent , Child , Child, Preschool , Female , Humans , Male , Motor Neuron Disease/diagnosis , Movement/physiology , Reproducibility of Results , Upper Extremity
12.
Sci Rep ; 9(1): 19972, 2019 12 30.
Article in English | MEDLINE | ID: mdl-31889062

ABSTRACT

The question whether novel rehabilitation interventions can exploit restorative rather than compensatory mechanisms has gained momentum in recent years. Assessments measuring selective voluntary motor control could answer this question. However, while current clinical assessments are ordinal-scaled, which could affect their sensitivity, lab-based assessments are costly and time-consuming. We propose a novel, interval-scaled, computer-based assessment game using low-cost accelerometers to evaluate selective voluntary motor control. Participants steer an avatar owl on a star-studded path by moving the targeted joint of the upper or lower extremities. We calculate a target joint accuracy metric, and an outcome score for the frequency and amplitude of involuntary movements of adjacent and contralateral joints as well as the trunk. We detail the methods and, as a first proof of concept, relate the results of select children with upper motor neuron lesions (n = 48) to reference groups of neurologically intact children (n = 62) and adults (n = 64). Linear mixed models indicated that the cumulative therapist score, rating the degree of selectivity, was a good predictor of the involuntary movements outcome score. This highlights the validity of this assessgame approach to quantify selective voluntary motor control and warrants a more thorough exploration to quantify changes induced by restorative interventions.


Subject(s)
Motor Neuron Disease/diagnosis , Psychomotor Performance/physiology , Range of Motion, Articular/physiology , Video Games , Accelerometry , Adolescent , Adult , Child , Disability Evaluation , Female , Humans , Male , Motor Neuron Disease/physiopathology
13.
J Neuroeng Rehabil ; 14(1): 87, 2017 08 30.
Article in English | MEDLINE | ID: mdl-28854939

ABSTRACT

BACKGROUND: Novel neurorehabilitation technologies build upon treatment principles derived from motor learning studies. However, few studies have investigated motor learning with assistive devices in children and adolescents with Cerebral Palsy (CP). The aim of this study was to investigate whether children with CP who trained with weight support in a playful, virtual environment would improve upper extremity task performance (i.e. skill acquisition), transfer, and retention, three aspects that indicate whether motor learning might have occurred or not. METHODS: Eleven children with CP (mean age 13.3 years, standard deviation 3.4 years), who were mildly to moderately impaired, participated. They played in the Armeo® Spring the exergame Moorhuhn with their more affected arm during 3 days (70 min pure play time). For this within-subject design, kinematic assessments, the Box and Block Test, and five items of the Melbourne Assessment were administered twice during a baseline week (one week before the intervention), directly before and after the intervention, and one day after the training phase (retention). RESULTS: The average exergame score improved from 209.55 to 339.73 (p < 0.001, Cohen's d = 1.80), indicating skill acquisition. The change in the Box and Block test improved from 0.45 (baseline week) to 3.95 (intervention week; p = 0.008, d = 1.59) indicating skill transfer. The kinematic assessments and the Melbourne items did not change. Improvement in game score and Box and Bock Test persisted one day later (retention). CONCLUSIONS: We found evidence indicating the successful acquisition, transfer, and retention of upper extremity skills in children with CP. We therefore infer that motor learning occurred when children with CP trained their more affected arm with weight-support in a playful, virtual environment.


Subject(s)
Cerebral Palsy/rehabilitation , Learning , Motor Skills/physiology , Upper Extremity , Adolescent , Arm , Biomechanical Phenomena , Child , Female , Humans , Male , Psychomotor Performance , Transfer, Psychology , Treatment Outcome
14.
J Cell Physiol ; 210(3): 740-9, 2007 Mar.
Article in English | MEDLINE | ID: mdl-17133351

ABSTRACT

Activating mutations of the RAS family of small GTPases are among the most common genetic events in human tumorigenesis. Constitutive activation of the three canonical family members, KRAS, NRAS, and HRAS segregate strongly by tissue type. Of these, KRAS mutations predominate in human tumors, including those arising from the colon and lung. We sought to compare the oncogenic contributions of different RAS isoforms in a comparable genetic setting and to explore downstream molecular changes that may explain the apparent differential oncogenic effects of the various RAS family members. We utilized colorectal cancer cell lines characterized by oncogenic KRAS in parallel with isogenically derived lines in which the mutant allele has been disrupted. We additionally attempted to reconstitute the isogenic derivatives with oncogenic forms of other RAS family members and analyze them in parallel. Pairwise analysis of HCT 116 and DLD-1 cell lines as well as their isogenic derivatives reveals distinct K-RAS(G13D) signatures despite the genetic similarities of these cell lines. In DLD-1, for example, oncogenic K-RAS enhances the motility of these cells by downregulation of Rap1 activity, yet is not associated with increased ERK1/2 phosphorylation. In HCT 116, however, ERK1/2 phosphorylation is elevated relative to the isogenic derivative, but Rap1 activity is unchanged. K-RAS is uniquely oncogenic in the colonic epithelium, though the molecular aspects of its oncogenic contribution are not necessarily conserved across cell lines. We therefore conclude that the oncogenic contribution of K-RAS is a function of its multifaceted functionality and is highly context-dependent.


Subject(s)
Colon/metabolism , Colorectal Neoplasms/genetics , Genes, ras/genetics , Intestinal Mucosa/metabolism , Cell Line, Tumor , Cell Transformation, Neoplastic/genetics , Colon/pathology , Colorectal Neoplasms/physiopathology , Gene Expression Regulation, Neoplastic , Genes, ras/physiology , Humans , Intestinal Mucosa/pathology , Isoenzymes , MAP Kinase Kinase Kinases/genetics , MAP Kinase Kinase Kinases/metabolism , Mitogen-Activated Protein Kinase 3/genetics , Mitogen-Activated Protein Kinase 3/metabolism , Signal Transduction/physiology , rap1 GTP-Binding Proteins/genetics , rap1 GTP-Binding Proteins/metabolism
15.
Proteomics ; 4(3): 793-811, 2004 Mar.
Article in English | MEDLINE | ID: mdl-14997500

ABSTRACT

Two-dimensional difference gel electrophoresis (2-D DIGE) coupled with mass spectrometry (MS) was used to investigate tumor-specific changes in the proteome of human colorectal cancers and adjacent normal mucosa. For each of six patients with different stages of colon cancer, Cy5-labeled proteins isolated from tumor tissue were combined with Cy3-labeled proteins isolated from neighboring normal mucosa and separated on the same 2-D gel along with a Cy2-labeled mixture of all 12 normal/tumor samples as an internal standard. Over 1500 protein spot-features were analyzed in each paired normal/tumor comparison, and using DIGE technology with the mixed-sample internal standard, statistically significant quantitative comparisons of each protein abundance change could be made across multiple samples simultaneously without interference due to gel-to-gel variation. Matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) and tandem (TOF/TOF) MS provided sensitive and accurate mass spectral data for database interrogation, resulting in the identification of 52 unique proteins (including redundancies due to proteolysis and post-translationally modified isoforms) that were changing in abundance across the cohort. Without the benefit of the Cy2-labeled 12 sample mixture internal standard, 42 of these proteins would have been overlooked due to the large degree of variation inherent between normal and tumor samples.


Subject(s)
Colonic Neoplasms/metabolism , Electrophoresis, Gel, Two-Dimensional/methods , Mass Spectrometry/methods , Proteome , Proteomics/methods , Adult , Aged , Aged, 80 and over , Databases as Topic , Female , Humans , Image Processing, Computer-Assisted , Intestinal Mucosa/pathology , Keratin-3 , Keratins/biosynthesis , Male , Middle Aged , Protein Isoforms , Sensitivity and Specificity , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods
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