ABSTRACT
PURPOSE: The aims of this study were to compare mechanical outputs (i.e. power and impulse), physiological (i.e. heart rate) and perceptual (i.e. effort and fatigue) responses in older men to traditional-set or different cluster-set configuration resistance training protocols. METHODS: In a randomized cross-over design, 20 healthy old men (aged 67.2 ± 2.1 years) completed four resistance training sessions using the back squat exercise loaded with optimal power loads. Training configurations were: traditional (TRA), three sets of six repetitions with 120-s rest between each set; Cluster-set 1 (CLU1), 24 single-repetition clusters with 10 s of rest after every cluster; Cluster-set 2 (CLU2), 12 double-repetition clusters with 20-s rest after every cluster; and Cluster-set 4 (CLU4), 6 quadruple-repetition clusters with 40-s rest after every cluster. RESULTS: Cluster-set configurations resulted in greater power outputs compared to traditional-set configuration (range 2.6-9.2%, all p [Formula: see text] 0.07 for main effect and protocol [Formula: see text] set interactions). CLU1 and CLU2 induced higher heart rate (range 7.1-10.5%, all p < 0.001 for main effect and protocol [Formula: see text] set interactions), lower rating of perceived exertion (range - 1.3 to - 3.2 AU, all p [Formula: see text] 0.006 for pairwise comparisons) and lower ratings of fatigue (range - 0.15 to - 4 AU, all p [Formula: see text] 0.012 for pairwise comparisons) compared to TRA and CLU4. Finally, an absolute preference for CLU2 was reported. CONCLUSIONS: Findings presented here support the prescription of CLU2 as an optimal resistance training configuration for trained older men using the back squat.
Subject(s)
Aging/physiology , Exercise Tolerance , Resistance Training/methods , Aged , Aged, 80 and over , Humans , Male , Muscle Contraction , Muscle Fatigue , Perception , Resistance Training/adverse effectsABSTRACT
New competencies may be learned through active experience (experiential learning or learning by doing) or observation of others' experiences (learning by observation). Observing another person performing a complex action facilitates the observer's acquisition of the same action. The present research is aimed at analyzing if the observation of specific explorative strategies adopted in a constrained environment, such as the Radial Arm Maze (RAM), could help young children to explore the maze and to build a cognitive spatial map of the explored environment. To this aim young children were randomly assigned to three groups: children who performed the RAM task following the observation of an actor solving the same maze by putting into action a highly structured exploratory strategy; children who performed the RAM task following the observation of the actor solving the same maze by putting into action a less structured exploratory strategy; children who directly performed the RAM task without any observation. The main result of the present research is that the children who observed the highly structured and correct exploratory strategy spent less time, made fewer errors, exhibited a longer spatial span, and thus they explored the maze more efficiently than the children who directly performed the RAM task without any observation. This finding indicates that when the observed explorative procedure is structured, sequential and repetitive the action understanding and information storage processes are more effective. Importantly, the observation of specific spatial strategies helped the children to build the cognitive spatial map of the explored environment and consequently to acquire/enrich the declarative knowledge of the environment.
Subject(s)
Child Behavior/physiology , Maze Learning/physiology , Social Learning/physiology , Animals , Child, Preschool , Female , Humans , MaleABSTRACT
This study aimed to investigate the effect of contact (C-SSG) and no-contact (NC-SSG) handball small-sided games (SSGs) on motion patterns and physiological responses of elite handball players. Twelve male handball players performed 10 C-SSG and 10 NC-SSG while being monitored through the heart rate (HR) and rate of perceived exertion (RPE) as physiological responses and time-motion activities profile using video-match analysis. Both game conditions resulted in similar HR responses (P > 0.05), but the NC-SSG led to a higher RPE scores. The time-motion activity analysis featured NC-SSG with a greater amount of walking (855.6 ± 25.1 vs. 690.6 ± 35.2 m) and backward movements (187.5 ± 12.3 vs. 142.5 ± 8.7 m) combined with fast running (232.3 ± 8.5 vs. 159.7 ± 5.7 m) and sprinting (79.5 ± 4.7 vs. 39.7 ± 3.7 m) activities (P < 0.001). Conversely, C-SSG had a higher percentage of jogging and sideway movements associated with greater frequency of jumping (0.87 ± 0.09 vs. 0.31 ± 0.06 nr) and physical contact (1.82 ± 0.55 vs. 0.25 ± 0.03 nr) events (P < 0.001). No between-regimen differences were found for the number of throws (P = 0.745). In addition, the RPE was significantly correlated with fast running relative distances (r = 0.909, P < 0.001) and sprinting relative distances (r = 0.939, P < 0.001). In conclusion, this investigation showed that both C-SSG and NC-SSG in team handball can effectively represent specifically oriented exercises, according to the sport-task and the performance demands.
Subject(s)
Athletic Performance/physiology , Sports/physiology , Competitive Behavior/physiology , Heart Rate/physiology , Humans , Male , Perception/physiology , Physical Conditioning, Human , Physical Exertion/physiology , Time and Motion Studies , Young AdultABSTRACT
Dello Iacono, A, Martone, D, Milic, M, and Padulo, J. Vertical- vs. horizontal-oriented drop jump training: chronic effects on explosive performances of elite handball players. J Strength Cond Res 31(4): 921-931, 2017-This study aimed to assess the chronic effects of vertical drop jump (VDJ)- and horizontal drop jump (HDJ)-based protocols on neuromuscular explosive abilities, such as jumping, sprinting, and changes of direction (COD). Eighteen elite male handball players (age 23.4 ± 4.6 years, height 192.5 ± 3.7 cm, weight 87.8 ± 7.4 kg) were assigned to either VDJ or HDJ group training twice a week for 10 weeks. Participants performed 5-8 sets × 6-10 repetitions of vertical alternate (VDJ) or horizontal alternate (HDJ) 1-leg drop jumps, landing from the top of a platform 25 cm in height. Before and after training, several performance, kinetic, and kinematic variables were assessed. The HDJ led to greater improvement of the sprint time (-8.5% vs. -4%, p ≤ 0.05) and COD performance in comparison with the VDJ (-7.9% vs. -1.1%, p ≤ 0.05), whereas the VDJ caused greater improvement in the vertical jump compared with the HDJ (+8.6% vs. +4.1%, p ≤ 0.05). Moreover, the VDJ regimen compared with the HDJ induced greater changes in the kinetic variables associated with vertical jumping performance, such as peak ground reaction forces (+10.3% vs. +4.3%), relative impulse (+12.4% vs. +5.7%), leg spring stiffness (+17.6% vs. +4.6%), contact time (CT) (-10.1% vs. -1.5%), and reactive strength index (+7.2% vs. +2.1%); all comparisons with p ≤ 0.05. Conversely, the HDJ regimen was able to improve the short-distance and COD performances by increasing the step length (+3.5% vs. +1.5% with p ≤ 0.05) and reducing the CT on COD (-12.1% vs. -2.1% with p ≤ 0.05) more than the VDJ. This investigation showed the crucial role that specific plyometric regimens play in optimizing similar biomechanical featured functional performances, such as jumping, sprinting, and COD.
Subject(s)
Athletes , Athletic Performance/physiology , Muscle Strength/physiology , Plyometric Exercise/methods , Sports , Adult , Biomechanical Phenomena , Exercise Test , Humans , Male , Young AdultABSTRACT
Dello Iacono, A, Martone, D, Cular, D, Milic, M, and Padulo, J. Game profile-based training in soccer: a new field approach. J Strength Cond Res 31(12): 3333-3342, 2017-The aim of the study was to profile and compare the time-motion, physiological, and neuromuscular responses of both National Youth League (NYL) and UEFA Youth League (UYL) matches with those of an experimental game profile-based training (GPBT) protocol. Time-motion traits and physiological, perceptual, and neuromuscular responses were investigated in 24 male soccer players across 14 matches and 6 GPBT training sessions, for a total of 420 samples. The GPBT had a greater influence on time-motion traits and perceptual responses than the NYL and UYL matches (all p < 0.001). No significant GPBT vs. match differences were found for mean heart rate or blood lactate (F = 1.228, p = 0.304, and F = 0.978, p = 0.385, respectively). Finally, the GPBT protocol led to greater impairment of the neuromuscular explosive performances when compared with those of the postmatch scores (squat jump: F = 19.991, p < 0.001; countermovement jump: F = 61.703, p < 0.001). Results identified the GPBT protocol as characterized by relatively greater high-intensity workloads than official NYL and UYL matches, requiring increased demanding efforts. In light of these outcomes, the GPBT protocol can be considered an advantageous training method for elite soccer players, capable of stimulating the physical effort and physiological capabilities required during a match. This approach is favorable when designing a training intervention according to the principle of sport specificity, as it is based on the specific metabolic demands.
Subject(s)
Athletic Performance/physiology , Exercise/physiology , Soccer/physiology , Adolescent , Heart Rate , Humans , Lactic Acid/blood , Male , Physical Exertion/physiology , Running/physiology , Time and Motion Studies , Young AdultABSTRACT
The aim of this study was to evaluate the effects of 6 different areas per player (AP) on exercise intensity (EI) measured during small-sided games (SSGs) and expressed as percentage of maximal heart rate (%MHR) and technical actions (TAs) involvement with the ball, crosses, headers, tackles, shots on goal, dribbling, passing, and target passing-in U-12 and U-14 soccer players during SSGs. Seventeen male U-12 soccer players (age 10.0 ± 0.5 years, body mass 39.3 ± 5.3 kg, and height 143.8 ± 4.6 cm) and 16 male U-14 soccer players (age 13.2 ± 0.3 years, body mass 46.6 ± 11.9 kg, and height 154.8 ± 8.5 cm) performed SSGs with different AP: 40, 50, 66.7, 90, 112.5, and 150 m. Our results indicate that at larger AP, the U-12 group's mean EI values were significantly higher than those at smaller AP (p ≤ 0.05); in addition, intergroup comparison showed that EI was higher in U-12 than that in U-14 players when AP of 112.5 and 150 m were considered (p ≤ 0.05). Technical action analysis evidenced that moving from smaller to larger AP, U-14 players adapted better to AP changes. In conclusion, these results suggest that AP influences differently EI and TAs in U-12 and U-14 players. Our results could be taken into account by conditioning coaches to better tailor the physiological and technical training in young players through the modulation of AP.
Subject(s)
Athletic Performance/physiology , Physical Conditioning, Human/methods , Soccer/physiology , Adolescent , Child , Heart Rate/physiology , Humans , MaleABSTRACT
Dello Iacono, A, Martone, D, and Padulo, J. Acute effects of drop-jump protocols on explosive performances of elite handball players. J Strength Cond Res 30(11): 3122-3133, 2016-This study aimed to assess the acute effects of vertical and horizontal drop jump-based postactivation potentiation (PAP) protocols on neuromuscular abilities in tasks such as jumping, sprinting, and change of direction (COD). Eighteen handball players were assessed before and after PAP regimens, consisting of either vertical single-leg drop-jumps (VDJ) or horizontal single-leg drop-jumps (HDJ) single-leg drop-jumps, on countermovement jump (CMJ), linear sprint, shuttle sprint, and agility performance. The HDJ led to greater improvement of the COD performance in comparison with the VDJ (-6.8 vs. -1.3%; p ≤ 0.05), whereas the VDJ caused greater improvement in the CMJ task compared with the HDJs (+6.5 vs. +1%; p ≤ 0.05). Moreover, the VDJ regimens compared with HDJ induced greater changes in most of the kinetic variables associated with vertical jumping performance, such as peak ground reaction forces (+9.6 vs. +1.3%), vertical displacement (-13.4 vs. -5.3%), leg-spring stiffness (+18.6 vs. +3.6%), contact time (-9.2 vs. -1.3%), and reactive strength index (+7.3 vs. +2.4%) (all comparisons with p ≤ 0.05). Conversely, the HDJ regimens were able to improve the COD performance only by reducing the contact time on COD more than the VDJ (-13.3 vs. -2.4% with p ≤ 0.05). The results showed that both PAPs were able to improve the performances that specifically featured similar force-orientation production. This investigation showed the crucial role that different and specific PAP regimens play in optimizing related functional performances. Specifically oriented vertical and horizontal single-leg drop-jump protocols represent viable means for achieving enhanced explosive-based tasks such as jumping and COD.
Subject(s)
Athletes , Athletic Performance/physiology , Exercise Test , Muscle Strength/physiology , Plyometric Exercise , Cross-Over Studies , Humans , Male , Random Allocation , Sports/physiology , Young AdultABSTRACT
Gene variants in MC4R, SIRT1 and FTO are associated with severe obesity and metabolic impairment in Caucasians. We investigated whether common variants in these genes are associated with metabolic syndrome (MetS) in a large group of morbidly obese young adults from southern Italy. One thousand morbidly obese subjects (62% women, mean body mass index 46.5 kg/m(2), mean age 32.6 years) whose families had lived in southern Italy for at least 2 generations were recruited. Single-nucleotide polymorphisms (SNPs) rs12970134, rs477181, rs502933 (MC4R locus), rs3818292, rs7069102, rs730821, rs2273773, rs12413112 (SIRT1 locus) and rs1421085, rs9939609, 9930506, 1121980 (FTO locus) were genotyped by Taqman assay; blood parameters were assayed by routine methods; the Fat Mass, Fat Free Mass, Respiratory Quotient, Basal Metabolic Rate (BMR) and waist circumference were also determined. Binomial logistic regression showed that the TA heterozygous genotype of SNP rs9939609 in the FTO gene was associated with the presence of MetS in our population [OR (95% CI): 2.53 (1.16-5.55)]. Furthermore, the FTO rs9939609 genotype accounted for 21.3% of the MetS phenotype together with total cholesterol, BMR and age. Our results extend the knowledge on genotype susceptibility for MetS in relation to a specific geographical area of residence.
Subject(s)
Metabolic Syndrome/complications , Metabolic Syndrome/genetics , Obesity, Morbid/genetics , Proteins/genetics , Adult , Alpha-Ketoglutarate-Dependent Dioxygenase FTO , Female , Genetic Association Studies , Genetic Variation , Humans , Italy , Logistic Models , Male , Metabolic Syndrome/pathology , Obesity, Morbid/pathology , Polymorphism, Single Nucleotide , Receptor, Melanocortin, Type 4/genetics , Sirtuin 1/geneticsABSTRACT
There are a lot of evidences on the beneficial effects mediated by exercise on the prevention of not communicable diseases (NCDs) including different type of cancer. The production of circulating exerkines transported in exosomes represents a novel pathway activated by exercise. However, the biological mechanisms that could explain the role of exosomes in cancer prevention have been not fully elucidated. The aim of this mini-review is to provide an update on the biological mechanisms bringing the release of muscle-derived exosomes during exercise and cancer prevention.
ABSTRACT
The effects of lifelong football training on bone health, body composition and physiological demands were evaluated. A total of 20 veteran football players (VPG; 73.4 ± 3.7 years) and 18 untrained age-matched men (CG; 75.6 ± 4.2 years) were enrolled. Whole-body and regional dual-energy X-ray absorptiometry scans of arms, legs, proximal femur and lower spine (L1-L4) were recorded in all participants. We observerd higher bone mineral density (BMD) in the whole-body, arms and femoral regions and higher bone mineral content (BMC) in the legs and lower spine compared to the CG (p < 0.05), also higher total lean body mass (p < 0.05) and lower total body fat percentage (p < 0.05), were found. No differences in food habits were evidenced between the VPG and the CG, as evaluated using 3-day food records. Resting heart rate (RHR), blood pressure (BP) and activity profile during a football match were recorded using a global positioning system only in the VPG. The mean heart rate (HR)of theoretical maximal HR (ThHRmax), and peak of ThHRmax were 83.9 ± 8.6% and 98.6 ± 10.2%, respectively; the mean of total distance covered was 3666 ± 721 m, and the means of accelerations and decelerations were 419 ± 61 and 428 ± 65, respectively. Lifelong participation in football training improves regional BMD and BMC in legs, femur and lumbar spine compared to the CG. A high number of intense actions in term of HR and accelerations and decelerations suggests an elevated energy expenditure that in turn correlates to the healthier body composition observed in the VPG compared to the CG.
ABSTRACT
Background: Physical fitness (PF) levels correlate with health hallmarks at all ages. In this study, w aimed to determine the PF level of schoolchildren from the Campania Region (Italy) through health-related PF (HRPF) components, taking into account body weight and sport practice (SP). Methods: PF level was determined in 565 schoolchildren aged 10−13 (11.7 ± 1.0 yrs; m: 353, f: 212) using some of the Eurofit battery tests. Results: 77% children practiced sport, boys more than girls (86% vs. 63%, respectively; p < 0.05). Boys performed better than girls (p < 0.05) in the Plate Tapping, Standing Broad Jump, Bent-Arm Hang, and 10 × 5 m Shuttle Run tests; girls performed better in the Sit-and-Reach Test (p < 0.05). Conclusion: Overweight/obese status negatively affects the muscular strength of lower limbs, even if it progressively improves during growth. SP was revealed to be a determinant in performance only in some PF tests, likely due to the heterogeneous training level among boys and girls who practice sport.
Subject(s)
Physical Fitness , Sports , Body Mass Index , Body Weight , Child , Exercise Test , Female , Humans , Male , OverweightABSTRACT
This study analyses the influence of different area per player (AP; 75, 98 and 131 m2) on the average metabolic power (MP) and other soccer-related performance variables in relation to the positional roles. We recruited 19 non-professional male soccer players (25.2 ± 6.3 y; 23.7 ± 2.3 kg/m2; 16.4 ± 6.3 y soccer experience) to play three different small-sided games (SSGs): SSG1 (5 vs. 5; 30 × 30 m; 5 min), SSG2 (5 vs. 5; 35 × 45 m; 5 min) and SSG3 (7 vs. 7; 35 × 45 m; 8 min). Specific playing rules were applied. GPS-assessed soccer-related variables were: average MP (AMP), distance covered in 1 min (DIS); % time spent at high speed (v > 16 km/h; % hst) or MP (>20 W/kg; % hmpt); % distance covered at high positive/negative speed (2 < v < 4 m/s2, % ACC; -6 < v < -2 m/s2, % DEC); and number of actions at high MP (hmpa). All recorded variables differed when each SSG was compared to the others (p < 0.05), but for hmpa for attackers. Most performance variables were positively associated with increasing AP (p < 0.05), but for % ACC and % DEC, and differed among positional roles within the same SSG (p < 0.05). Here the general applicability of SSGs, regardless the physical/technical skills of the group of players, to enhance performance is confirmed; furthermore, quantitative advices on AMP and other performance variables are provided to achieve significant improvements in all soccer players of the team.
Subject(s)
Athletic Performance , Blood Group Antigens , Running , Soccer , Humans , Male , Pilot ProjectsABSTRACT
Anabolic androgenic steroids, a class of steroid hormones related to testosterone, are natural ligands of androgen receptor (AR), a member of the nuclear receptor superfamily of ligand-activated transcription factors. AR binds specific DNA elements, known as androgen-response elements. Testosterone, the main male sexual hormone, binds AR directly and indirectly, through conversion into dihydrotestosterone (DHT), its more active metabolite. Anabolic androgenic steroids are frequently detected in the urine of doped athletes; their consumption is also growing among sport amateurs and adolescents. The effects of androgens can differ depending on the target cells and/or tissues. To gain insight into transcription activation mechanisms of AR, we investigated AR protein signaling in human peripheral blood lymphocytes treated with supraphysiological doses of DHT. We performed a comparative proteomic analysis and we identified about 30 differentially expressed proteins. At least five species contained a consensus androgen-response elements sequence in the promoter region of related coding genes. The analysis also revealed that high doses of DHT activate the drug detoxification process, could stimulate an increase in cell motility and exert a prosurvival effect rather than an apoptotic one.
Subject(s)
Dihydrotestosterone/pharmacology , Leukocytes, Mononuclear/metabolism , Proteome/drug effects , Proteomics/methods , Receptors, Androgen/metabolism , Signal Transduction/drug effects , Adult , Apoptosis/drug effects , Caspase 3/metabolism , Computational Biology , Electrophoresis, Gel, Two-Dimensional , Humans , Leukocytes, Mononuclear/drug effects , Male , Proteome/chemistry , Proteome/metabolism , Reproducibility of Results , Response ElementsABSTRACT
BACKGROUND: The usefulness of adapted small-sided games (SSGs) in improving cardiac function in subjects with T2DM is still debated. Here we evaluated the effects of 18 weeks indoor muscular activation training (6 weeks; IMA) followed by adapted SSGs football training (12 weeks) on cardiac function, muscular fitness, body composition and adiponectin expression in sedentary T2DM volunteers. METHODS: Six T2DM patients underwent IMA protocol of 6 weeks, twice a week followed by 12 weeks SSGs (5-a-side, once a week) training. Glucose, lipid profile and serum homocysteine concentration, body composition (BC), bone mineral density (DEXA), were determined at baseline and after 18 weeks (IMA+SSGs). VO
Subject(s)
Cardiorespiratory Fitness/physiology , Diabetes Mellitus, Type 2/physiopathology , Soccer/physiology , Adiponectin/blood , Body Composition , Exercise Test/methods , Football , Humans , Male , Middle Aged , Oxygen Consumption/physiologyABSTRACT
Vitamin D is thought to regulate skeletal muscle function and boost physical performance. The aim of this study was to assess the relationship between vitamin D and physical performance in physically active children. This cross-sectional study included 125 children who practice football as a leisure activity. Plasma 25-hydroxyvitamin D (25-OHD) was assessed using a chemiluminescence immunoassay method. Vitamin D inadequacy was defined as 25-OHD < 20 ng/mL. Physical performance testing included measurements of muscle strength (maximal isometric contraction), jumping ability (vertical jump, standing broad jump, triple hop test), linear sprint (10 m and 20 m), and agility (9 × 4-m shuttle run). Plasma 25-OHD concentrations were positively correlated with muscle strength (r = 0.539; p < 0.001), vertical jump (r = 0.528; p < 0.001), and standing broad jump (r = 0.492; p < 0.001) but inversely correlated with sprint performance (r = -0.539; p < 0.001). In multivariate analysis models, plasma 25-OHD concentrations were associated with each physical performance parameter independently of age, maturity status, body mass index, fat mass, and protein and calcium intakes. In conclusion, a low plasma 25-OHD level was associated with decreased muscle strength, agility, and jumping and sprinting abilities in physically active children. Vitamin D inadequacy may limit exercise performance. Further research should verify whether correction of vitamin D deficiency enhances physical performance.
Subject(s)
25-Hydroxyvitamin D 2/blood , Athletic Performance , Calcifediol/blood , Muscle Strength , Muscle Weakness/etiology , Physical Fitness , Vitamin D Deficiency/physiopathology , Adolescent , Adolescent Behavior , Child , Child Behavior , Confounding Factors, Epidemiologic , Cross-Sectional Studies , Healthy Lifestyle , Humans , Male , Motor Activity , Prevalence , Soccer , Sports and Recreational Facilities , Track and Field , Tunisia/epidemiology , Vitamin D Deficiency/blood , Vitamin D Deficiency/epidemiologyABSTRACT
Performance-enhancing drugs (PEDs) gained wide popularity not only among sportsmen but also among specific subsets of population, such as adolescents. Apart from their claimed effects on athletic performance, they are very appealing due to the body shaping effect exerted on fat mass and fat-free mass. Besides the "underestimated" massive misuse of PEDs, the short- as well as long-term consequences of such habits remain largely unrecognized. They have been strictly associated with serious adverse effects, but molecular mechanisms are yet to be elucidated. Here, we analyze the current understanding of the molecular effects of supraphysiological doses of doping agents in healthy biological systems, at genomic and proteomic levels, in order to define the molecular sensors of organ/tissue impairment, determined by their misuse. The focus is put on the anabolic androgenic steroids (AASs), specifically testosterone (T) and its most potent derivative dihydrotestosterone (DHT), and on the peptide hormones, specifically the growth hormone (GH) and the insulin-like growth factor-1 (IGF-1). A map of molecular targets is defined and the risk incidence for human health is taken into account.
Subject(s)
Doping in Sports , Growth Hormone/pharmacology , Testosterone/pharmacology , Apoptosis/drug effects , Biomarkers/analysis , Biomarkers/chemistry , Growth Hormone/adverse effects , Humans , Proteomics , Signal Transduction/drug effects , Testosterone/adverse effectsABSTRACT
The melanocortin-4 receptor (MC4R) is involved in regulating energy homeostasis; mutations in this gene have been associated with 1-5% of early-onset human obesity. The aim of this study was to functionally characterize MC4R mutations identified in morbidly obese subjects living in Southern Italy. We studied their ligand binding, signaling pathway and subcellular localization. As expected, mutants Q43X and S19fsX51, which produce truncated forms of receptor, were devoid of activity. The activity of mutants W174C and A175T were very different even though the mutations are adjacent and are in the same transmembrane helix (TMH). In fact, the production and expression of mutant A175T on the plasma-membrane (PM) was similar to that of the wild-type (wt) receptor and the mutant retained 70% of wt receptor activity; on the contrary, the production of W174C mutant in the cytoplasm was similar to that of the wt receptor and mutant A175T but was only barely detectable on the PM and was devoid of activity. Confocal microscopy showed that W174C remained entrapped in the endoplasmic reticulum (ER) of the cells. Structural analysis showed that substitution of Trp174, located in the middle of TMH4 and 100% conserved in all known MC4Rs, with Cys could impair the relative orientation of TMH2 and TMH4 thereby affecting the overall protein architecture. Furthermore, co-expression studies showed that mutant A175T but not W174C had a dominant negative effect on the wt receptor activity.