Do patients with well-functioning total hip arthroplasty achieve typical sagittal plane hip kinematics? A proof of concept study.

BACKGROUND: Total hip arthroplasty (THA) patients have been shown to not achieve normal sagittal plane hip kinematics. However, previous studies have only conducted group level analysis and as such lack the sensitivity to highlight whether individual patients do achieve normal hip kinematics. As such this study looked to determine whether some patients with well-functioning THA achieve typical sagittal plane hip kinematics. METHODS: Sagittal plane hip kinematics were collected on 11 well-functioning THA patients (Oxford Hip Score = 46 ± 3) and 10 asymptomatic controls using a 3-dimensional motion analysis system during self-paced walking. High-functioning THA patients were identified as those who displayed sagittal plane hip kinematics that were within the variance of the control group on average, and low-functioning patients as those who did not. RESULTS: 5 THA patients were identified as high-functioning, displaying hip kinematics within the variance of the control group. High-functioning THA patients displayed peak hip flexion and extension values more closely aligned to asymptomatic control group than low-functioning patients. However, hip range of motion was comparable between high- and low-functioning total hip arthroplasty patients and reduced compared to controls. CONCLUSION: The presence of high-functioning THA patients who display comparable sagittal plane hip kinematics to controls suggests these patients do achieve normative function and challenges the conclusions of previous group level analysis. Understanding why some patients achieve better function post-operatively will aid pre- and post-operative practices to maximise functional recovery.

Acute physiological, endocrine, biochemical and performance responses associated with amateur boxing: A systematic review with meta-analysis.

Previous research has explored the demands of amateur boxing-specific activity; however, no holistic review of the acute responses to such activity currently exists. This paper aimed to provide a systematic review of the available literature on the acute physiological, endocrine, biochemical, and performance responses to amateur boxing-specific activity. Following a search of EBSCOhost, SPORTDiscus, PubMed and Google Scholar databases, 25 studies were identified as meeting the inclusion criteria for the review. The methodological quality of the included studies was assessed via a modified Downs and Black checklist. Random-effects meta-analysis of standardised mean differences (SMD) revealed large (SMD = 4.62) increases in pre-post blood lactate (BLa), cortisol (SMD = 1.33), myoglobin (Mb) (SMD = 1.43), aspartate transaminase (AST) (SMD = 1.37), and alanine aminotransferase (ALT) (SMD = 0.97), in addition to moderate increases in creatine kinase (CK) (SMD = 0.65). Small pre-post increases in counter-movement jump (CMJ) height (SMD = 0.33) were observed. Consistently greater pre-post alterations were observed in competitive bouts, followed by sparring, and less so in boxing-specific simulations. Considerable physiological, endocrine, and biochemical responses are elicited following amateur boxing. Interestingly, neuromuscular and task-specific performance may not deteriorate following boxing-specific activity. The findings of the review may assist in the designing and periodising of boxing-specific training, dependent on the desired physical adaptations, training phase, and recovery status of the amateur boxer.HighlightsAmateur boxing elicits a considerable acute physiological, hormonal, and biochemical response.Such responses are typically greater in competitive bouts, followed by sparring, and less so in simulated activity.The considerable demands of amateur boxing-specific activity do not appear to negatively affect neuromuscular or task-specific performance.Amateur boxers may be conditioned to preserve performance despite the acute demands of the sport, or the lack of performance decrement may reflect the short duration of amateur boxing.

A commentary on soccer match-play simulations for applied research and practice.

Soccer is a fast-growing area of research, demonstrated by a 10-fold increase in the number of PubMed articles derived from the search term 'soccer' between 2001 and 2021. The scope of contemporary soccer-related articles ranges from match-play observations to laboratory evaluations of performance. The activity profile of soccer match-play is variable and techniques to collect data within matches are limited. Soccer-specific simulations have been developed to simulate the evolving demands of match-play. The evolutionary designs of novel simulations provide a reproducible exercise stimulus for varying researcher and practitioner objectives. The applied researcher can utilise simulations to investigate the efficacy of nutritional interventions and environmental stress on performance, while assessing the physiological and biomechanical responses to representations of match-play. Practitioners can adopt simulations for rehabilitation to progressively facilitate return-to-play processes, while implementing extra top-up conditioning sessions for unused and partial-match players. However, there are complexities involved with the selection of varying simulations which are dependent on the research question or practical application. There also remains a paucity of published information to support researchers and practitioners in selecting from differing simulation models. To assist with researcher and practitioner interpretations, we present a commentary of the current simulations to inform decision-making processes for research and training purposes and enhance the application of future research. An objective scoring system was adopted for rating the research and practical applications of each simulation design. Overall scores of 22, 16 and 18 out of 36 were revealed for free-running (n = 7), non-motorised- (n = 4) and motorised-treadmill-based simulations (n = 4), respectively.

Postactivation Performance Enhancement of Amateur Boxers' Punch Force and Neuromuscular Performance Following 2 Upper-Body Conditioning Activities.

PURPOSE: The purpose of this study was to assess the efficacy of upper-body punch-specific isometric (ISO) and elastic resistance (ER) conditioning activities (CAs) on the punch force and neuromuscular performance of amateur boxers. METHODS: Ten male senior elite amateur boxers (19.7 [1.2]| y; height 180.9 [7.0] cm; mass 78.7 [9.6] kg) visited the laboratory on 4 separate occasions. Initially, the participants performed baseline physical tests comprising bench-press 1-repetition maximum and countermovement jumps. On the other 3 occasions, the boxers performed maximal punches against a vertically mounted force plate and maximal countermovement jumps prior to and following an ISO or ER CA, as well as a control trial. RESULTS: No interactions between CA × time were found in all performance variables. As observed by mean changes, effect sizes, and signal:noise ratio, both the ISO and ER, but not the control trial, consistently produced small to moderate, worthwhile increases in punch force and rate of force development, with the greatest increases in performance typically observed in the ISO trial. No meaningful improvements were observed in countermovement jump performance in all trials, indicative of a localized postactivation performance enhancement effect. CONCLUSION: In conclusion, the ISO and ER CAs may be implemented in an amateur boxers' warm-up to acutely enhance punch-force variables, although the ISO punch appears to be the superior CA to improve punch-specific performance. The CAs used in the present study may also be relevant to other combat sports inclusive of a striking element.

Effect of alterations in whole-body cryotherapy (WBC) exposure on post-match recovery markers in elite Premier League soccer players.

The use of whole-body cryotherapy (WBC) as a recovery intervention is prevalent amongst elite soccer players. However, there is a distinct lack of data available around chronic WBC use and post-match recovery markers in elite soccer. The aim of this study was to investigate the impact of different levels of WBC exposure on subjective and objective measures of post-match recovery in elite soccer players during a chronic exposure period. Sixteen male senior professional outfield soccer players participated in this study over two seasons. K means cluster analysis was used to classify low (-114 ± 2°C for 133 ± 2 s), medium (-121 ± 1°C for 173 ± 2 s) and high (-133 ± 1°C for 181 ± 2 s) cryotherapy exposure indexes (CEI). Salivary markers (immunoglobulin A (IgA) and alpha amylase (AA)) and subjective wellness scores (perceived fatigue, sleep quality, general muscle soreness and stress) were collected post-match across both seasons. Training load (session-RPE) was collected and used as a covariate to control for the load amongst groups. No differences were seen in perceived measures of wellness and salivary AA. Significantly lower IgA concentrations were observed in the medium CEI group (255 ± 32 µg∙ml-1) compared to the low (328 ± 38 µg∙ml-1) and high (306 ± 32 µg∙ml-1) groups. Therefore, increasing the level of chronic WBC exposure appears to have no additional benefit on subjective recovery and alpha amylase response post-match. However, there appears to be an optimal chronic WBC dose with regards to IgA response.

Maturity-associated considerations for training load, injury risk, and physical performance in youth soccer: One size does not fit all.

Biological maturation can be defined as the timing and tempo of progress to achieving a mature state. The estimation of age of peak height velocity (PHV) or percentage of final estimated adult stature attainment (%EASA) is typically used to inform the training process in young athletes. In youth soccer, maturity-related changes in anthropometric and physical fitness characteristics are diverse among individuals, particularly around PHV. During this time, players are also at an increased risk of sustaining an overuse or growth-related injury. As a result, the implementation of training interventions can be challenging. The purpose of this review was to (1) highlight and discuss many of the methods that can be used to estimate maturation in the applied setting and (2) discuss the implications of manipulating training load around PHV on physical development and injury risk. We have provided key stakeholders with a practical online tool for estimating player maturation status (Supplementary Maturity Estimation Tools). Whilst estimating maturity using predictive equations is useful in guiding the training process, practitioners should be aware of its limitations. To increase the accuracy and usefulness of data, it is also vital that sports scientists implement reliable testing protocols at predetermined time-points.

The effects of medium chain triglyceride (MCT) supplementation using a C8:C10 ratio of 30:70 on cognitive performance in healthy young adults.

PURPOSE: The brain can utilise medium chain triglycerides (MCTs) as an alternative fuel to glucose, and research has shown that MCT ingestion improves cognitive function in diseased and/or elderly individuals. The aim of this study is to determine if these improvements can also be observed in young, healthy adults. Furthermore, we aim to establish the ideal dosage and timeframe necessary for an effect. METHODS: Participants were divided equally into three groups of 10 (Placebo (0 g), 12 g and 18 g MCT/day) and were supplemented for 4 weeks. The supplement had a C8:C10 ratio of 30:70. Participants visited the laboratory once a week for 5 weeks (baseline, test weeks 1-4) to undergo a battery of cognitive tests; Trail Making, Digit Span, Spatial Span, Covert Shift of Attention, and Rapid Visual Information Processing. RESULTS: After 2-3 weeks of supplementation, MCT ingestion enhanced performance in cognitive tasks, including: Trail Making A/B and Digit Span Forwards/Backwards (ps<0.001) when compared to a placebo group taking a carbohydrate gel. In Spatial Span Backwards, there was a significant main effect of group (p = 0.002). Where significance was seen, there were main effects of time after 2-3 weeks (ps<0.05). There was minimal difference between the two MCT intervention groups in most measures (ps>0.05). There were also null results in tasks measuring attention and reaction time (ps>0.05). CONCLUSIONS: MCT ingestion improved cognitive performance after 2-3 weeks, with minimal difference between taking 12 g and 18 g MCT/day groups, suggesting a possible dose-response threshold at 12 g MCT/day when supplementing over a short period.

Angle-Specific Isokinetic Metrics Highlight Strength Training Needs of Elite Youth Soccer Players.

Eustace, SJ, Page, RM, and Greig, M. angle specific isokinetic metrics highlight strength training needs of elite youth soccer players. J Strength Cond Res 34(11): 3258-3265, 2020-The purpose of this study was to assess traditional and angle-specific isokinetic strength of eccentric knee flexors (eccKFs) and concentric knee extensors (conKEs) between senior professional and youth soccer players. Thirty-four male soccer players (17 senior and 17 youth) were recruited for bilateral assessments at 180, 270, and 60°·s. Peak torque (PT), dynamic control ratio (DCR), angle of peak torque (APT), functional range (FR), angle-specific torque (AST), and angle-specific DCR (DCRAST) were compared. The eccentric knee flexor (eccKF) and conKE PT (p = 0.782) and DCR (p = 0.508) were not different between groups across all angular velocities. Significant differences were identified for eccKF APT (p = 0.018) and FR (p = 0.006), DCRAST at 270°·s (p = 0.031), and in AST data recorded across angular velocities for eccKF and conKE (p = 0.003). Traditional strength measures were not sensitive to playing age, with implications for misinterpretation in training prescription. By contrast, AST data did differentiate between ages. Strength deficits that highlight the muscle contraction type, angular velocity, and joint angle can be manipulated within an individualized training intervention. Given the relevance to injury etiology, this study highlights potential implications for improved assessment strategies to inform training prescription for performance and injury prevention. Given the high number of injuries in adolescent soccer players, and in line with previous recommendations, practitioners should consider using more informed and specific strength and conditioning practices at younger ages.

Physical Response to Pad- and Bag-Based Boxing-Specific Training Modalities.

Finlay, MJ, Greig, M, McCarthy, J, and Page, RM. Physical response to pad- and bag-based boxing-specific training modalities. J Strength Cond Res 34(4): 1052-1061, 2020-This study examined the differences in the physical response elicited from a contemporary Boxing-Specific Exercise Protocol (BSEP) performed using a punch bag and a pad routine. Fourteen male elite amateur boxers (age = 22 ± 2 years; height = 176.9 ± 7.3 cm; body mass = 78.8 ± 8.7 kg; and V[Combining Dot Above]O2max = 55.94 ± 5.96 ml·kg·min) were recruited. The BSEP comprised 3 × 3-minute rounds. Average (HRave) and peak (HRpeak) heart rate, average (V[Combining Dot Above]O2ave) and peak oxygen consumption (V[Combining Dot Above]O2peak), blood lactate (BLa) concentrations, rating of perceived exertion (RPE), and both triaxial and uniaxial PlayerLoad metrics were recorded during each trial. The PlayerLoad metrics were recorded at both the cervical and lumbar spine. BLa increased significantly across rounds, with higher values recorded in the pad trial (pad = 2.7 ± 0.8 mmol·L; bag = 2.3 ± 0.9 mmol·L). A similar response was also identified for the HRave (pad = 160 ± 9 b·min; bag = 150 ± 16 b·min) and V[Combining Dot Above]O2ave data (pad = 38.00 ± 0.31 ml·kg·min; bag = 34.40 ± 1.06 ml·kg·min). A significant main effect for time was also recorded for the RPE data; however, there were no significant differences between trials. Conversely, the triaxial (PLTotal) and medial-lateral (PLML) data were higher in the punch bag trial. There was also a main effect for time for all the PlayerLoad metrics. PLTotal, PLML, and vertical PlayerLoad were significantly higher in the lumbar region when compared with the cervical region. With implications for boxing-specific conditioning, the pad routine was more physiologically demanding, but less mechanically demanding than the bag routine.

Physical Response to a Simulated Period of Soccer-Specific Fixture Congestion.

Page, RM, Marrin, K, Brogden, CM, and Greig, M. Physical response to a simulated period of soccer-specific fixture congestion. J Strength Cond Res 33(4): 1075-1085, 2019-The aim of this study was to assess the physiological, perceptual, and mechanical measures associated with the completion of a simulated period of short-term soccer-specific fixture congestion. Ten male semiprofessional soccer players completed 3 trials of a treadmill-based match simulation, with 48 hours interspersing each trial. A repeated measures general linear model identified significantly (p = 0.02) lower knee flexor peak torque (PT) recorded at 300°·s in the second (141.27 ± 28.51 N·m) and third trials (139.12 ± 26.23 N·m) when compared with the first trial (154.17 ± 35.25 N·m). Similarly, muscle soreness (MS) and PT data recorded at 60°·s were significantly (p ≤ 0.05) different in the third trial (MS = 42 ± 25 a.u; PT60 = 131.10 ± 35.38 N·m) when compared with the first trial (MS = 29 ± 29 a.u; PT60 = 145.61 ± 42.86 N·m). Significant (p = 0.003) differences were also observed for mean electromyography (EMGmean) of bicep femoris between the third trial (T0-15 = 126.36 ± 15.57 µV; T75-90 = 52.18 ± 17.19 µV) and corresponding time points in the first trial (T0-15 = 98.20 ± 23.49 µV; T75-90 = 99.97 ± 39.81 µV). Cumulative increases in perceived exertion, heart rate, oxygen consumption, blood lactate concentrations, EMGmean, and PlayerLoad (PL) were recorded across each trial. Muscle soreness and PT were also significantly different after trial. There were, however, no significant main effects or interactions for the salivary immunoglobulin A and the medial-lateral PL metrics. These data suggest a biomechanical and muscular emphasis with residual fatigue, with implications for injury risk and the development of recovery strategies.

Quantifying the Physical Response to a Contemporary Amateur Boxing Simulation.

Finlay, MJ, Greig, M, and Page, RM. Quantifying the physical response to a contemporary amateur boxing simulation. J Strength Cond Res 32(4): 1005-1012, 2018-This study examined the physical response to a contemporary boxing-specific exercise protocol (BSEP), based on notational analysis of amateur boxing. Nine male senior elite amateur boxers completed a 3 × 3-minute BSEP, with a 1-minute passive recovery period interspersing each round. Average (HRave) and peak (HRpeak) heart rates, average (V[Combining Dot Above]O2ave) and peak oxygen consumptions (V[Combining Dot Above]O2peak), blood lactate (BLa) concentrations, rating of perceived exertion, and both triaxial and uniaxial PlayerLoad metrics were recorded during the completion of the BSEP. Blood lactate concentration increased significantly in each round (Round 1 = 2.4 ± 1.3 mmol·L; Round 2 = 3.3 ± 1.7 mmol·L; Round 3 = 4.3 ± 2.6 mmol·L). Significantly lower HRave and HRpeak values were found in the first round (HRave: 150 ± 15 b·min; HRpeak: 162 ± 12 b·min) when compared with the second (HRave: 156 ± 16 b·min; HRpeak: 166 ± 13 b·min) and third (HRave: 150 ± 15 b·min; HRpeak: 169 ± 14 b·min). No significant differences were found in any of the V[Combining Dot Above]O2 or PlayerLoad metrics recorded during the BSEP. The BSEP based on notational analysis elicited a fatigue response across rounds, confirming its validity. The BSEP can be used as a training tool for boxing-specific conditioning with implications for reduced injury risk, and to assess the physical response to boxing-specific interventions. Moreover, the BSEP can also be manipulated to suit all levels of participants or training phases, with practical applications in performance monitoring and microcycle periodization.

Biomechanical and Physiological Response to a Contemporary Soccer Match-Play Simulation.

The intermittent activity profile of soccer match play increases the complexity of the physical demands. Laboratory models of soccer match play have value in controlled intervention studies, developed around manipulations of the activity profile to elicit a desired physiological or biomechanical response. Contemporary notational analyses suggest a profile comprising clusters of repeat sprint efforts, with implications for both biomechanical and physiological load. Eighteen male soccer players completed a 90-minute treadmill protocol based on clusters of repeat sprint efforts. Each 15-minute bout of exercise was quantified for uniaxial (medial-lateral [PLML], anterior-posterior [PLAP], and vertical [PLV]) and triaxial PlayerLoad (PLTotal). The relative contributions of the uniaxial PlayerLoad vectors (PLML%, PLAP%, and PLV%) were also examined. In addition to rating of perceived exertion, the physiological response comprised heart rate, blood lactate concentration, and both peak and average oxygen consumption. Triaxial PlayerLoad increased (p = 0.02) with exercise duration (T0-15 = 206.26 ± 14.37 a.u. and T45-60 = 214.51 ± 14.97 a.u.) and remained elevated throughout the second half. This fatigue effect was evident in both the PLML and PLAP movement planes. The mean relative contributions of PLV%:PLAP%:PLML% were consistent at ∼48:28:23. The physiological response was comparable with match play, and a similar magnitude of increase at ∼5% was observed in physiological parameters. Changes in PlayerLoad might reflect a change in movement quality with fatigue, with implications for both performance and injury risk, reflecting observations of match play. The high frequency of speed change elicits a 23% contribution from mediolateral load, negating the criticism of treadmill protocols as "linear."

Suppression of Aß toxicity by puromycin-sensitive aminopeptidase is independent of its proteolytic activity.

The accumulation of ß-amyloid (Aß) peptide in the brain is one of the pathological hallmarks of Alzheimer's disease and is thought to be of primary aetiological significance. In an unbiased genetic screen, we identified puromycin-sensitive aminopeptidase (PSA) as a potent suppressor of Aß toxicity in a Drosophila model system. We established that coexpression of Drosophila PSA (dPSA) in the flies' brains improved their lifespan, protected against locomotor deficits, and reduced brain Aß levels by clearing the Aß plaque-like deposits. However, confocal microscopy and subcellular fractionation of amyloid-expressing 7PA2 cells demonstrated that PSA localizes to the cytoplasm. Therefore, PSA and Aß are unlikely to be in the same cellular compartment; moreover, when we artificially placed them in the same compartment in flies, we could not detect a direct epistatic interaction. The consequent hypothesis that PSA's suppression of Aß toxicity is indirect was supported by the finding that Aß is not a proteolytic substrate for PSA in vitro. Furthermore, we showed that the enzymatic activity of PSA is not required for rescuing Aß toxicity in neuronal SH-SY5Y cells. We investigated whether the stimulation of autophagy by PSA was responsible for these protective effects. However PSA's promotion of autophagosome fusion with lysosomes required proteolytic activity and so its effect on autophagy is not identical to its protection against Aß toxicity.

Loss of PAFAH1B2 reduces amyloid-ß generation by promoting the degradation of amyloid precursor protein C-terminal fragments.

Amyloid-ß peptide (Aß) is believed to play a central role in the pathogenesis of Alzheimer's disease. In view of the side effects associated with inhibiting the secretases that produce Aß, new molecular targets are required to provide alternative therapeutic options. We used RNA interference (RNAi) to systematically screen the Drosophila genome to identify genes that modulate Aß production upon knockdown. RNAi of 41 genes in Drosophila cells significantly lowered Aß without affecting general secretion or viability. After the γ-secretase complex components, the most potent effect was observed for platelet activating factor acetylhydrolase α (Paf-AHα), and, in mammalian cells, the effect was replicated for its ortholog PAFAH1B2. Knockdown of PAFAH1B2 strongly reduced Aß secretion from human cells, and this effect was confirmed in primary cells derived from PAFAH1B2 knock-out mice. Reduced Aß production was not attributable to altered ß-amyloid precursor protein (APP) ectodomain shedding but was a result of an enhanced degradation of APP C-terminal fragments (CTFs) in the absence of PAFAH1B2 but not its close homolog PAFAH1B3. Enhanced degradation of APP CTFs was selective because no such effects were obtained for Notch or E-/N-cadherin. Thus, we have identified an important protein that can selectively modify Aß generation via a novel mechanism, namely enhanced degradation of its immediate precursor. In view of the absence of a neurological phenotype in PAFAH1B2 knock-out mice, targeted downregulation of PAFAH1B2 may be a promising new strategy for lowering Aß.

Attenuated Abeta42 responses to low potency gamma-secretase modulators can be overcome for many pathogenic presenilin mutants by second-generation compounds.

Sequential processing of the ß-amyloid precursor protein by ß- and γ-secretase generates the amyloid ß-peptide (Aß), which is widely believed to play a causative role in Alzheimer disease. Selective lowering of the pathogenic 42-amino acid variant of Aß by γ-secretase modulators (GSMs) is a promising therapeutic strategy. Here we report that mutations in presenilin (PS), the catalytic subunit of γ-secretase, display differential responses to non-steroidal anti-inflammatory drug (NSAID)-type GSMs and more potent second-generation compounds. Although many pathogenic PS mutations resisted lowering of Aß(42) generation by the NSAID sulindac sulfide, the potent NSAID-like second-generation compound GSM-1 was capable of lowering Aß(42) for many but not all mutants. We further found that mutations at homologous positions in PS1 and PS2 can elicit differential Aß(42) responses to GSM-1, suggesting that a positive GSM-1 response depends on the spatial environment in γ-secretase. The aggressive pathogenic PS1 L166P mutation was one of the few pathogenic mutations that resisted GSM-1, and Leu-166 was identified as a critical residue with respect to the Aß(42)-lowering response of GSM-1. Finally, we found that GSM-1-responsive and -resistant PS mutants behave very similarly toward other potent second-generation compounds of different structural classes than GSM-1. Taken together, our data show that a positive Aß(42) response for PS mutants depends both on the particular mutation and the GSM used and that attenuated Aß(42) responses to low potency GSMs can be overcome for many PS mutants by second generation GSMs.

Bepridil and amiodarone simultaneously target the Alzheimer's disease beta- and gamma-secretase via distinct mechanisms.

The two proteases beta-secretase and gamma-secretase generate the amyloid beta peptide and are drug targets for Alzheimer's disease. Here we tested the possibility of targeting the cellular environment of beta-secretase cleavage instead of the beta-secretase enzyme itself. beta-Secretase has an acidic pH optimum and cleaves the amyloid precursor protein in the acidic endosomes. We identified two drugs, bepridil and amiodarone, that are weak bases and are in clinical use as calcium antagonists. Independently of their calcium-blocking activity, both compounds mildly raised the membrane-proximal, endosomal pH and inhibited beta-secretase cleavage at therapeutically achievable concentrations in cultured cells, in primary neurons, and in vivo in guinea pigs. This shows that an alkalinization of the cellular environment could be a novel therapeutic strategy to inhibit beta-secretase. Surprisingly, bepridil and amiodarone also modulated gamma-secretase cleavage independently of endosomal alkalinization. Thus, both compounds act as dual modulators that simultaneously target beta- and gamma-secretase through distinct molecular mechanisms. In addition to Alzheimer's disease, compounds with dual properties may also be useful for drug development targeting other membrane proteins.

Beta-amyloid precursor protein mutants respond to gamma-secretase modulators.

Pathogenic generation of the 42-amino acid variant of the amyloid beta-peptide (Abeta) by beta- and gamma-secretase cleavage of the beta-amyloid precursor protein (APP) is believed to be causative for Alzheimer disease (AD). Lowering of Abeta(42) production by gamma-secretase modulators (GSMs) is a hopeful approach toward AD treatment. The mechanism of GSM action is not fully understood. Moreover, whether GSMs target the Abeta domain is controversial. To further our understanding of the mode of action of GSMs and the cleavage mechanism of gamma-secretase, we analyzed mutations located at different positions of the APP transmembrane domain around or within the Abeta domain regarding their response to GSMs. We found that Abeta(42)-increasing familial AD mutations of the gamma-secretase cleavage site domain responded robustly to Abeta(42)-lowering GSMs, especially to the potent compound GSM-1, irrespective of the amount of Abeta(42) produced. We thus expect that familial AD patients carrying mutations at the gamma-secretase cleavage sites of APP should respond to GSM-based therapeutic approaches. Systematic phenylalanine-scanning mutagenesis of this region revealed a high permissiveness to GSM-1 and demonstrated a complex mechanism of GSM action as other Abeta species (Abeta(41), Abeta(39)) could also be lowered besides Abeta(42). Moreover, certain mutations simultaneously increased Abeta(42) and the shorter peptide Abeta(38), arguing that the proposed precursor-product relationship of these Abeta species is not general. Finally, mutations of residues in the proposed GSM-binding site implicated in Abeta(42) generation (Gly-29, Gly-33) and potentially in GSM-binding (Lys-28) were also responsive to GSMs, a finding that may question APP substrate targeting of GSMs.

Microglial Cx3cr1 knockout prevents neuron loss in a mouse model of Alzheimer's disease.

Microglia, the immune cells of the brain, can have a beneficial effect in Alzheimer's disease by phagocytosing amyloid-beta. Two-photon in vivo imaging of neuron loss in the intact brain of living Alzheimer's disease mice revealed an involvement of microglia in neuron elimination, indicated by locally increased number and migration velocity of microglia around lost neurons. Knockout of the microglial chemokine receptor Cx3cr1, which is critical in neuron-microglia communication, prevented neuron loss.

Fenton chemistry and oxidative stress mediate the toxicity of the beta-amyloid peptide in a Drosophila model of Alzheimer's disease.

The mechanism by which aggregates of the beta-amyloid peptide (Abeta) mediate their toxicity is uncertain. We show here that the expression of the 42-amino-acid isoform of Abeta (Abeta(1-42)) changes the expression of genes involved in oxidative stress in a Drosophila model of Alzheimer's disease. A subsequent genetic screen confirmed the importance of oxidative stress and a molecular dissection of the steps in the cellular metabolism of reactive oxygen species revealed that the iron-binding protein ferritin and the H(2)O(2) scavenger catalase are the most potent suppressors of the toxicity of wild-type and Arctic (E22G) Abeta(1-42). Likewise, treatment with the iron-binding compound clioquinol increased the lifespan of flies expressing Arctic Abeta(1-42). The effect of iron appears to be mediated by oxidative stress as ferritin heavy chain co-expression reduced carbonyl levels in Abeta(1-42) flies by 65% and restored the survival and locomotion function to normal. This was achieved despite the presence of elevated levels of the Abeta(1-42). Taken together, our data show that oxidative stress, probably mediated by the hydroxyl radical and generated by the Fenton reaction, is essential for Abeta(1-42) toxicity in vivo and provide strong support for Alzheimer's disease therapies based on metal chelation.

Generation of Abeta38 and Abeta42 is independently and differentially affected by familial Alzheimer disease-associated presenilin mutations and gamma-secretase modulation.

Alzheimer disease amyloid beta-peptide (Abeta) is generated via proteolytic processing of the beta-amyloid precursor protein by beta- and gamma-secretase. Gamma-secretase can be blocked by selective inhibitors but can also be modulated by a subset of non-steroidal anti-inflammatory drugs, including sulindac sulfide. These drugs selectively reduce the generation of the aggregation-prone 42-amino acid Abeta(42) and concomitantly increase the levels of the rather benign Abeta(38). Here we show that Abeta(42) and Abeta(38) generation occur independently from each other. The amount of Abeta(42) produced by cells expressing 10 different familial Alzheimer disease (FAD)-associated mutations in presenilin (PS) 1, the catalytic subunit of gamma-secretase, appeared to correlate with the respective age of onset in patients. However, Abeta(38) levels did not show a negative correlation with the age of onset. Modulation of gamma-secretase activity by sulindac sulfide reduced Abeta(42) in the case of wild type PS1 and two FAD-associated PS1 mutations (M146L and A285V). The remaining eight PS1 FAD mutants showed either no reduction of Abeta(42) or only rather subtle effects. Strikingly, even the mutations that showed no effect on Abeta(42) levels allowed a robust increase of Abeta(38) upon treatment with sulindac sulfide. Similar observations were made for fenofibrate, a compound known to increase Abeta(42) and to decrease Abeta(38). For mutants that predominantly produce Abeta(42), the ability of fenofibrate to further increase Abeta(42) levels became diminished, whereas Abeta(38) levels were altered to varying extents for all mutants analyzed. Thus, we conclude that Abeta(38) and Abeta(42) production do not depend on each other. Using an independent non-steroidal anti-inflammatory drug derivative, we obtained similar results for PS1 as well as for PS2. These in vitro results were confirmed by in vivo experiments in transgenic mice expressing the PS2 N141I FAD mutant. Our findings therefore have strong implications on the selection of transgenic mouse models used for screening of the Abeta(42)-lowering capacity of gamma-secretase modulators. Furthermore, human patients with certain PS mutations may not respond to gamma-secretase modulators.