Sleeping Beauty mRNA-LNP enables stable rAAV transgene expression in mouse and NHP hepatocytes and improves vector potency.

Recombinant adeno-associated virus (rAAV) vector gene delivery systems have demonstrated great promise in clinical trials but continue to face durability and dose-related challenges. Unlike rAAV gene therapy, integrating gene addition approaches can provide curative expression in mitotically active cells and pediatric populations. We explored a novel in vivo delivery approach based on an engineered transposase, Sleeping Beauty (SB100X), delivered as an mRNA within a lipid nanoparticle (LNP), in combination with an rAAV-delivered transposable transgene. This combinatorial approach achieved correction of ornithine transcarbamylase deficiency in the neonatal Spfash mouse model following a single delivery to dividing hepatocytes in the newborn liver. Correction remained stable into adulthood, while a conventional rAAV approach resulted in a return to the disease state. In non-human primates, integration by transposition, mediated by this technology, improved gene expression 10-fold over conventional rAAV-mediated gene transfer while requiring 5-fold less vector. Additionally, integration site analysis confirmed a random profile while specifically targeting TA dinucleotides across the genome. Together, these findings demonstrate that transposable elements can improve rAAV-delivered therapies by lowering the vector dose requirement and associated toxicity while expanding target cell types.

Neuromechanical Differences between Pronated and Supinated Forearm Positions during Upper-Body Wingate Tests.

Arm-cycling is a versatile exercise modality with applications in both athletic enhancement and rehabilitation, yet the influence of forearm orientation remains understudied. Thus, this study aimed to investigate the impact of forearm position on upper-body arm-cycling Wingate tests. Fourteen adult males (27.3 ± 5.8 years) underwent bilateral assessments of handgrip strength in standing and seated positions, followed by pronated and supinated forward arm-cycling Wingate tests. Electromyography (EMG) was recorded from five upper-extremity muscles, including anterior deltoid, triceps brachii lateral head, biceps brachii, latissimus dorsi, and brachioradialis. Simultaneously, bilateral normal and propulsion forces were measured at the pedal-crank interface. Rate of perceived exertion (RPE), power output, and fatigue index were recorded post-test. The results showed that a pronated forearm position provided significantly (p < 0.05) higher normal and propulsion forces and triceps brachii muscle activation patterns during arm-cycling. No significant difference in RPE was observed between forearm positions (p = 0.17). A positive correlation was found between seated handgrip strength and peak power output during the Wingate test while pronated (dominant: p = 0.01, r = 0.55; non-dominant: p = 0.03, r = 0.49) and supinated (dominant: p = 0.03, r = 0.51; don-dominant: p = 0.04, r = 0.47). Fatigue changed the force and EMG profile during the Wingate test. In conclusion, this study enhances our understanding of forearm position's impact on upper-body Wingate tests. These findings have implications for optimizing training and performance strategies in individuals using arm-cycling for athletic enhancement and rehabilitation.

Motor unit firing rates during slow and fast contractions in boys and men.

BACKGROUND: Motor unit (MU) activation during maximal contractions is lower in children compared with adults. Among adults, discrete MU activation differs, depending on the rate of contraction. We investigated the effect of contraction rate on discrete MU activation in boys and men. METHODS: Following a habituation session, 14 boys and 20 men completed two experimental sessions for knee extension and wrist flexion, in random order. Maximal voluntary isometric torque (MVIC) was determined before completing trapezoidal isometric contractions (70%MVIC) at low (10%MVIC/s) and high (35%MVIC/s) contraction rates. Surface electromyography was captured from the vastus lateralis (VL) and flexor carpi radialis (FCR) and decomposed into individual MU action potential (MUAP) trains. RESULTS: In both groups and muscles, the initial MU firing rate (MUFR) was greater (p < 0.05) at high compared with low contraction rates. The increase in initial MUFR at the fast contraction in the VL was greater in men than boys (p < 0.05). Mean MUFR was significantly lower during fast contractions only in the FCR (p < 0.05). In both groups and muscles, the rate of decay of MUFR with increasing MUAP amplitude was less steep (p < 0.05) during fast compared with slow contractions. CONCLUSION: In both groups and muscles, initial MUFRs, as well as MUFRs of large MUs were higher during fast compared with slow contractions. However, in the VL, the increase in initial MUFR was greater in men compared with boys. This suggests that in large muscles, men may rely more on increasing MUFR to generate torque at faster rates compared with boys.

Genetic analysis and natural history of Parkinson's disease due to the LRRK2 G2019S variant.

The LRRK2 G2019S variant is the most common cause of monogenic Parkinson's disease (PD); however, questions remain regarding the penetrance, clinical phenotype and natural history of carriers. We performed a 3.5-year prospective longitudinal online study in a large number of 1286 genotyped LRRK2 G2019S carriers and 109 154 controls, with and without PD, recruited from the 23andMe Research Cohort. We collected self-reported motor and non-motor symptoms every 6 months, as well as demographics, family histories and environmental risk factors. Incident cases of PD (phenoconverters) were identified at follow-up. We determined lifetime risk of PD using accelerated failure time modelling and explored the impact of polygenic risk on penetrance. We also computed the genetic ancestry of all LRRK2 G2019S carriers in the 23andMe database and identified regions of the world where carrier frequencies are highest. We observed that despite a 1 year longer disease duration (P = 0.016), LRRK2 G2019S carriers with PD had similar burden of motor symptoms, yet significantly fewer non-motor symptoms including cognitive difficulties, REM sleep behaviour disorder (RBD) and hyposmia (all P-values ≤ 0.0002). The cumulative incidence of PD in G2019S carriers by age 80 was 49%. G2019S carriers had a 10-fold risk of developing PD versus non-carriers. This rose to a 27-fold risk in G2019S carriers with a PD polygenic risk score in the top 25% versus non-carriers in the bottom 25%. In addition to identifying ancient founding events in people of North African and Ashkenazi descent, our genetic ancestry analyses infer that the G2019S variant was later introduced to Spanish colonial territories in the Americas. Our results suggest LRRK2 G2019S PD appears to be a slowly progressive predominantly motor subtype of PD with a lower prevalence of hyposmia, RBD and cognitive impairment. This suggests that the current prodromal criteria, which are based on idiopathic PD, may lack sensitivity to detect the early phases of LRRK2 PD in G2019S carriers. We show that polygenic burden may contribute to the development of PD in the LRRK2 G2019S carrier population. Collectively, the results should help support screening programmes and candidate enrichment strategies for upcoming trials of LRRK2 inhibitors in early-stage disease.

Translation initiation factor eIF1.2 promotes Toxoplasma stage conversion by regulating levels of key differentiation factors.

The parasite Toxoplasma gondii persists in its hosts by converting from replicating tachyzoites to latent bradyzoites housed in tissue cysts. The molecular mechanisms that mediate T. gondii differentiation remain poorly understood. Through a mutagenesis screen, we identified translation initiation factor eIF1.2 as a critical factor for T. gondii differentiation. A F97L mutation in eIF1.2 or the genetic ablation of eIF1.2 (∆eif1.2) markedly impeded bradyzoite cyst formation in vitro and in vivo. We demonstrated, at single-molecule level, that the eIF1.2 F97L mutation impacts the scanning process of the ribosome preinitiation complex on a model mRNA. RNA sequencing and ribosome profiling experiments unveiled that ∆eif1.2 parasites are defective in upregulating bradyzoite induction factors BFD1 and BFD2 during stress-induced differentiation. Forced expression of BFD1 or BFD2 significantly restored differentiation in ∆eif1.2 parasites. Together, our findings suggest that eIF1.2 functions by regulating the translation of key differentiation factors necessary to establish chronic toxoplasmosis.

mRNA cap-binding protein eIF4E1 is a novel regulator of Toxoplasma gondii latency.

The protozoan parasite Toxoplasma gondii causes serious opportunistic disease due to its ability to persist in patients as latent tissue cysts. The molecular mechanisms coordinating conversion between proliferative parasites (tachyzoites) and latent cysts (bradyzoites) are not fully understood. We previously showed that phosphorylation of eIF2α accompanies bradyzoite formation, suggesting that this clinically relevant process involves regulation of mRNA translation. In this study, we investigated the composition and role of eIF4F multi-subunit complexes in translational control. Using CLIPseq, we find that the cap-binding subunit, eIF4E1, localizes to the 5'-end of all tachyzoite mRNAs, many of which show evidence of stemming from heterogeneous transcriptional start sites. We further show that eIF4E1 operates as the predominant cap-binding protein in two distinct eIF4F complexes. Using genetic and pharmacological approaches, we found that eIF4E1 deficiency triggers efficient spontaneous formation of bradyzoites without stress induction. Consistent with this result, we also show that stress-induced bradyzoites exhibit reduced eIF4E1 expression. Overall, our findings establish a novel role for eIF4F in translational control required for parasite latency and microbial persistence. IMPORTANCE: Toxoplasma gondii is an opportunistic pathogen important to global human and animal health. There are currently no chemotherapies targeting the encysted form of the parasite. Consequently, a better understanding of the mechanisms controlling encystation is required. Here we show that the mRNA cap-binding protein, eIF4E1, regulates the encystation process. Encysted parasites reduce eIF4E1 levels, and depletion of eIF4E1 decreases the translation of ribosome-associated machinery and drives Toxoplasma encystation. Together, these data reveal a new layer of mRNA translational control that regulates parasite encystation and latency.

Impact of repetitive mouse clicking on forearm muscle fatigue and mouse aiming performance.

Exercise induced performance fatigue has been shown to impair many aspects of fine motor function in the distal upper limb. However, most fatiguing protocols do not reflect the conditions experienced with computer use. The purpose of this study was to determine how a prolonged, low-force mouse clicking fatigue protocol impacts performance fatigue of the distal upper limb for gamers and non-gamers. Participants completed a total of 1 h of mouse clicking at 5 clicks per second. Muscle fatigue and performance were intermittently assessed. RMS amplitude increased for the forearm flexors throughout the fatigue protocol. Accuracy decreased following the first bout of clicking and returned to baseline values after 40-min. EDC and ECU displayed the greatest muscle activity while aiming, producing 11.4% and 12.9% of MVC, respectively. These findings indicate that mouse clicking may not result in performance fatigue, however, high levels of extensor activity may explain common injuries among gamers.

Robotic systems for upper-limb rehabilitation in multiple sclerosis: a SWOT analysis and the synergies with virtual and augmented environments.

The robotics discipline is exploring precise and versatile solutions for upper-limb rehabilitation in Multiple Sclerosis (MS). People with MS can greatly benefit from robotic systems to help combat the complexities of this disease, which can impair the ability to perform activities of daily living (ADLs). In order to present the potential and the limitations of smart mechatronic devices in the mentioned clinical domain, this review is structured to propose a concise SWOT (Strengths, Weaknesses, Opportunities, and Threats) Analysis of robotic rehabilitation in MS. Through the SWOT Analysis, a method mostly adopted in business management, this paper addresses both internal and external factors that can promote or hinder the adoption of upper-limb rehabilitation robots in MS. Subsequently, it discusses how the synergy with another category of interaction technologies - the systems underlying virtual and augmented environments - may empower Strengths, overcome Weaknesses, expand Opportunities, and handle Threats in rehabilitation robotics for MS. The impactful adaptability of these digital settings (extensively used in rehabilitation for MS, even to approach ADL-like tasks in safe simulated contexts) is the main reason for presenting this approach to face the critical issues of the aforementioned SWOT Analysis. This methodological proposal aims at paving the way for devising further synergistic strategies based on the integration of medical robotic devices with other promising technologies to help upper-limb functional recovery in MS.

Sex differences in strength at the shoulder: a systematic review.

Background: Understanding differential strength capability between sexes is critical in ergonomics and task design. Variations in study designs and outcome measures generates challenges in establishing workplace guidelines for strength requirements to minimize upper extremity risk for workers. The purpose of this systematic review was to collate and summarize sex differences in strength at the shoulder across movement directions and contraction types. Methods: A total of 3,294 articles were screened from four databases (Embase, Medline, SCOPUS, and Web of Science). Eligibility criteria included observational studies, direct measurement of muscular joint, and healthy adult participants (18-65 years old). Strength outcome measures were normalized to percentages of male outputs to allow comparisons across articles. Results: A total of 63 studies were included within the final review. Majority of articles observed increased strength in males; the gap between male-female strength was greater in flexion and internal/external rotation, with females generating ~30% of male strength; scaption strength ratios were most consistent of the movement groups, with females generating 55-62% of male strength. Conclusion: Sex strength differences should be considered as an important factor for workplace task design as women are more at risk for occupational-related injuries than men in equivalent strength requirements. Differences in strength were not synonymous across motions; females demonstrated increased disparity relative to male strength in horizontal flexion/extension, forward flexion and internal/external rotation. Some movements had an extremely limited pool of available studies for examination which identified critical research gaps within the literature. Collating and quantifying strength differences is critical for effective workstation design with a range of users to mitigate potential overexertion risk and musculoskeletal injury.

Post-invasion selection acts on standing genetic variation despite a severe founding bottleneck.

Invasive populations often have lower genetic diversity relative to the native-range populations from which they derive.1,2 Despite this, many biological invaders succeed in their new environments, in part due to rapid adaptation.3,4,5,6 Therefore, the role of genetic bottlenecks in constraining the adaptation of invaders is debated.7,8,9,10 Here, we use whole-genome resequencing of samples from a 10-year time-series dataset, representing the natural invasion of the Asian honey bee (Apis cerana) in Australia, to investigate natural selection occurring in the aftermath of a founding event. We find that Australia's A. cerana population was founded by as few as one colony, whose arrival was followed by a period of rapid population expansion associated with an increase of rare variants.11 The bottleneck resulted in a steep loss of overall genetic diversity, yet we nevertheless detected loci with signatures of positive selection during the first years post-invasion. When we investigated the origin of alleles under selection, we found that selection acted primarily on the variation introduced by founders and not on the variants that arose post-invasion by mutation. In all, our data highlight that selection on standing genetic variation can occur in the early years post-invasion, even where founding bottlenecks are severe.

Developmental changes in motor unit activity patterns: child-adult comparison using discrete motor unit analysis.

Using global surface electromyography (sEMG) and the sEMG threshold it has been suggested that children activate their type-II motor unit (MU) to a lesser extent compared with adults. However, when age-related differences in discrete MU activation are examined using sEMG decomposition this phenomenon is not observed. Furthermore, findings from these studies are inconsistent and conflicting. Therefore, the purpose of this study was to examine differences in discrete MU activation of the vastus lateralis (VL) between boys and men during moderate-intensity knee extensions. Seventeen boys and 20 men completed two laboratory sessions. Following a habituation session, maximal voluntary isometric knee extension (MVIC) torque was determined before completing trapezoidal contractions at 70% MVIC. sEMG of the VL was captured and mathematically decomposed into individual MU action potential trains. Motor unit action potential amplitude (MUAPamp), recruitment threshold (RT), and MU firing rates (MUFR) were calculated. We observed that MUAPamp-RT slope was steeper in men compared with boys (p < 0.05) even after accounting for fat thickness and quadriceps muscle depth. The mean MUFR and y-intercept of the MUFR-RT relationship were significantly (p < 0.001) lower in boys than in men. The slope of the MUFR-RT relationship tended to be steeper in men, but the differences did not reach statistical significance (p = 0.056). Overall, our results suggest that neural strategies used to produce torque are different among boys and men. Such differences may be related, in part, to boys' lower MUFR and lesser ability to activate their higher-threshold MUs. Although, other factors (e.g., muscle composition) likely also play a role.

The Effects of Load, Crank Position, and Sex on the Biomechanics and Performance during an Upper Body Wingate Anaerobic Test.

INTRODUCTION: The upper body Wingate Anaerobic Test (WAnT) is a 30-s maximal effort sprint against a set load (percentage of body mass). However, there is no consensus on the optimal load and no differential values for males and females, even when there are well-studied anatomical and physiological differences in muscle mass for the upper body. Our goal was to describe the effects of load, sex, and crank position on the kinetics, kinematics, and performance of the upper body WAnT. METHODS: Eighteen participants (9 females) performed three WAnTs at 3%, 4%, and 5% of body mass. Arm crank forces, 2D kinematics, and performance variables were recorded during each WAnT. RESULTS: Our results showed an increase of ~49% effective force, ~36% peak power, ~5° neck flexion, and ~30° shoulder flexion from 3% to 5% load ( P < 0.05). Mean power and anaerobic capacity decreased by 15%, with no changes in fatigue index ( P < 0.05). The positions of higher force efficiency were at 12 and 6 o'clock. The least force efficiency occurred at 3 o'clock ( P < 0.05). Sex differences showed that males produced 97% more effective force and 109% greater mean power than females, with 11.7% more force efficiency ( P < 0.001). Males had 16° more head/neck flexion than females, and females had greater elbow joint variability with 17° more wrist extension at higher loads. Males cycled ~32% faster at 3% versus 5% WAnT load with a 65% higher angular velocity than females. Grip strength, maximal voluntary isometric contraction, mass, and height positively correlated with peak and mean power ( P < 0.001). CONCLUSIONS: In conclusion, load, sex, and crank position have a significant impact on performance of the WAnT. These factors should be considered when developing and implementing an upper body WAnT.

Different discrete motor-unit activation patterns in the flexor carpi radialis in boys and men.

BACKGROUND: Lower activation of higher threshold (type-II) motor units (MUs) has been suggested in children compared with adults. We examined child-adult differences in discrete MU activation of the flexor carpi radialis (FCR). METHODS: Fifteen boys (10.2 ± 1.4 years), and 17 men (25.0 ± 2.7 years) completed 2 laboratory sessions. Following a habituation session, maximal voluntary isometric wrist flexion torque (MVIC) was determined before completing trapezoidal isometric contractions at 70%MVIC. Surface electromyography was captured by Delsys Trigno Galileo sensors and decomposed into individual MU action potential trains. Recruitment threshold (RT), and MU firing rates (MUFR) were calculated. RESULTS: MVIC was significantly greater in men (10.19 ± 1.92 Nm) than in boys (4.33 ± 1.47 Nm) (p < 0.05), but not statistically different after accounting for differences in body size. Mean MUFR was not different between boys (17.41 ± 7.83 pps) and men (17.47 ± 7.64 pps). However, the MUFR-RT slope was significantly (p < 0.05) steeper (more negative) in boys, reflecting a progressively greater decrease in MUFR with increasing RT. Additionally, boys recruited more of their MUs early in the ramped contraction. CONCLUSION: Compared with men, boys tended to recruit their MUs earlier and at a lower percentage of MVIC. This difference in MU recruitment may explain the greater decrease in MUFR with increasing RT in boys compared with men. Overall, these findings suggest an age-related difference in the neural strategy used to develop moderate-high torque in wrist flexors, where boys recruit more of their MUs earlier in the force gradation process, possibly resulting in a narrower recruitment range.

PCSK9 genetic variants and risk of vascular and non-vascular diseases in Chinese and UK populations.

AIMS: Lowering low-density lipoprotein cholesterol (LDL-C) through PCSK9 inhibition represents a new therapeutic approach to preventing and treating cardiovascular disease (CVD). Phenome-wide analyses of PCSK9 genetic variants in large biobanks can help to identify unexpected effects of PCSK9 inhibition. METHODS AND RESULTS: In the prospective China Kadoorie Biobank, we constructed a genetic score using three variants at the PCSK9 locus associated with directly measured LDL-C [PCSK9 genetic score (PCSK9-GS)]. Logistic regression gave estimated odds ratios (ORs) for PCSK9-GS associations with CVD and non-CVD outcomes, scaled to 1 SD lower LDL-C. PCSK9-GS was associated with lower risks of carotid plaque [n = 8340 cases; OR = 0.61 (95% confidence interval: 0.45-0.83); P = 0.0015], major occlusive vascular events [n = 15 752; 0.80 (0.67-0.95); P = 0.011], and ischaemic stroke [n = 11 467; 0.80 (0.66-0.98); P = 0.029]. However, PCSK9-GS was also associated with higher risk of hospitalization with chronic obstructive pulmonary disease [COPD: n = 6836; 1.38 (1.08-1.76); P = 0.0089] and with even higher risk of fatal exacerbations amongst individuals with pre-existing COPD [n = 730; 3.61 (1.71-7.60); P = 7.3 × 10-4]. We also replicated associations for a PCSK9 variant, reported in UK Biobank, with increased risks of acute upper respiratory tract infection (URTI) [pooled OR after meta-analysis of 1.87 (1.38-2.54); P = 5.4 × 10-5] and self-reported asthma [pooled OR of 1.17 (1.04-1.30); P = 0.0071]. There was no association of a polygenic LDL-C score with COPD hospitalization, COPD exacerbation, or URTI. CONCLUSION: The LDL-C-lowering PCSK9 genetic variants are associated with lower risk of subclinical and clinical atherosclerotic vascular disease but higher risks of respiratory diseases. Pharmacovigilance studies may be required to monitor patients treated with therapeutic PCSK9 inhibitors for exacerbations of respiratory diseases or respiratory tract infections. LAY SUMMARY: Genetic analyses of over 100 000 participants of the China Kadoorie Biobank, mimicking the effect of new drugs intended to reduce cholesterol by targeting the PCSK9 protein, have identified potential severe effects of lower PCSK9 activity in patients with existing respiratory disease.PCSK9 genetic variants that are associated with lower cholesterol and reduced rates of cardiovascular disease are also associated with increased risk of a range of respiratory diseases, including asthma, upper respiratory tract infections, and hospitalization with chronic obstructive pulmonary disease (COPD).These genetic variants are not associated with whether or not individuals have COPD; instead, they are specifically associated with an increase in the chance of those who already have COPD being hospitalized and even dying, suggesting that careful monitoring of such patients should be considered during development of and treatment with anti-PCSK9 medication.

Time-varying changes in median nerve deformation and position in response to quantified pinch and grip forces.

The ability of the median nerve (MN) to adapt in response to altered carpal tunnel conditions is important to mitigate compressive stress on the nerve. We assessed changes in MN deformation and position throughout the entire time course of hand force exertions. Fourteen right-handed participants ramped up force from 0% to 50% of maximal voluntary force (MVF) before ramping force back down in three different hand force exertion tasks (pulp pinch, chuck pinch, power grip). Pinch and grip forces were measured with a digital dynamometer, which were time synchronized with transverse carpal tunnel images obtained via ultrasound. Ultrasound images were extracted in 10% increments between 0% and 50% MVF while ramping force up (loading phase) and down (unloading phase). MN deformation and position relative to the flexor digitorum superficialis tendon of the long finger were assessed in concert. During loading, the nerve became more circular while displacing dorsally and ulnarly. These changes primarily occurred at the beginning of the hand force exertions while ramping force up from 0% to 20%, with very little change between 20% and 50% MVF. Interestingly, deformation and position changes during loading were not completely reversed during unloading while ramping force down. These findings indicate an initial reorganization of carpal tunnel structures. Mirrored changes in nerve deformation and position may also reflect strain-related characteristics of adjoining subsynovial connective tissue. Regardless, time-varying changes in nerve deformation and position appear to be an important accommodative mechanism in the healthy carpal tunnel in response to gripping and pinching tasks.

Sex differences in wrist strength: a systematic review.

Sex differences in strength have been attributed to differences in body anthropometrics and composition; these factors are often ignored when generating workplace guidelines. These differences directly impact the upper extremity, leaving female workers exposed to injury risk. The wide range of tools and techniques for measuring upper extremity strength presents a challenge to ergonomists and work task designers; collating outcomes to provide a clear outlook of differences between males and females is essential and the purpose of this work. Four online databases were searched (PROSPERO ID: CRD42022339023) with a focus on articles assessing sex differences in wrist strength. A total of 2,378 articles were screened for relevancy; 25 full-text articles were included in this systematic review. Articles examined movement pairs (ulnar/radial deviation, pronation/supination, and flexion/extension), as well as contraction types (isometric and isokinetic) to observe sex differences in wrist strength. Across all articles, females produced ∼60-65% of male flexion/extension strength, ∼55-60% pronation/supination strength, and ∼60-70% ulnar/radial deviation strength. Overall, females presented lower strength-producing abilities than males, but when considering strength relative to body mass, male-female differences were less pronounced and occasionally females surpassed male strength metrics; typically, this occurred during flexion/extension, particularly in isokinetic contractions. This review has identified a scarcity of articles examining ulnar/radial deviation, pronation/supination, as well as isokinetic contractions; these are needed to supplement workplace exposure guidelines.

Translation initiation factor eIF1.2 promotes Toxoplasma stage conversion by regulating levels of key differentiation factors.

The parasite Toxoplasma gondii persists in its hosts by converting from replicating tachyzoites to latent bradyzoites housed in tissue cysts. The molecular mechanisms that mediate T. gondii differentiation remain poorly understood. Through a mutagenesis screen, we identified translation initiation factor eIF1.2 as a critical factor for T. gondii differentiation. A F97L mutation in eIF1.2 or the genetic ablation of eIF1.2 (Δ eIF1.2 ) markedly impeded bradyzoite cyst formation in vitro and in vivo . We demonstrated, at single-molecule level, that the eIF1.2 F97L mutation impacts the scanning process of the ribosome preinitiation complex on a model mRNA. RNA sequencing and ribosome profiling experiments unveiled that Δ eIF1.2 parasites are defective in the upregulating bradyzoite induction factors BFD1 and BFD2 during stress-induced differentiation. Forced expression of BFD1 or BFD2 significantly restored differentiation in Δ eIF1.2 parasites. Together, our findings suggest that eIF1.2 functions by regulating the translation of key differentiation factors necessary to establish chronic toxoplasmosis.

Plasma Proteomics to Identify Drug Targets for Ischemic Heart Disease.

BACKGROUND: Integrated analyses of plasma proteomic and genetic markers in prospective studies can clarify the causal relevance of proteins and discover novel targets for ischemic heart disease (IHD) and other diseases. OBJECTIVES: The purpose of this study was to examine associations of proteomics and genetics data with IHD in population studies to discover novel preventive treatments. METHODS: We conducted a nested case-cohort study in the China Kadoorie Biobank (CKB) involving 1,971 incident IHD cases and 2,001 subcohort participants who were genotyped and free of prior cardiovascular disease. We measured 1,463 proteins in the stored baseline samples using the OLINK EXPLORE panel. Cox regression yielded adjusted HRs for IHD associated with individual proteins after accounting for multiple testing. Moreover, cis-protein quantitative loci (pQTLs) identified for proteins in genome-wide association studies of CKB and of UK Biobank were used as instrumental variables in separate 2-sample Mendelian randomization (MR) studies involving global CARDIOGRAM+C4D consortium (210,842 IHD cases and 1,378,170 controls). RESULTS: Overall 361 proteins were significantly associated at false discovery rate <0.05 with risk of IHD (349 positively, 12 inversely) in CKB, including N-terminal prohormone of brain natriuretic peptide and proprotein convertase subtilisin/kexin type 9. Of these 361 proteins, 212 had cis-pQTLs in CKB, and MR analyses of 198 variants in CARDIOGRAM+C4D identified 13 proteins that showed potentially causal associations with IHD. Independent MR analyses of 307 cis-pQTLs identified in Europeans replicated associations for 4 proteins (FURIN, proteinase-activated receptor-1, Asialoglycoprotein receptor-1, and matrix metalloproteinase-3). Further downstream analyses showed that FURIN, which is highly expressed in endothelial cells, is a potential novel target and matrix metalloproteinase-3 a potential repurposing target for IHD. CONCLUSIONS: Integrated analyses of proteomic and genetic data in Chinese and European adults provided causal support for FURIN and multiple other proteins as potential novel drug targets for treatment of IHD.

mRNA cap-binding protein eIF4E1 is a novel regulator of Toxoplasma gondii latency.

The protozoan parasite Toxoplasma gondii causes serious opportunistic disease due to its ability to persist in patients as latent tissue cysts. The molecular mechanisms coordinating conversion between proliferative parasites (tachyzoites) and dormant cysts (bradyzoites) are not fully understood. We previously showed that phosphorylation of eIF2α accompanies bradyzoite formation, suggesting that this clinically relevant process involves regulation of mRNA translation. In this study, we investigated the composition and role of eIF4F multi-subunit complexes in translational control. Using CLIPseq, we find that the cap-binding subunit, eIF4E1, localizes to the 5'-end of all tachyzoite mRNAs, many of which show evidence of stemming from heterogenous transcriptional start sites. We further show that eIF4E1 operates as the predominant cap-binding protein in two distinct eIF4F complexes. Using genetic and pharmacological approaches, we found that eIF4E1 deficiency triggers efficient spontaneous formation of bradyzoites without stress induction. Consistent with this result, we also show that stress-induced bradyzoites exhibit reduced eIF4E1 expression. Overall, our findings establish a novel role for eIF4F in translational control required for parasite latency and microbial persistence.

Central Adiposity Increases Risk of Kidney Stone Disease through Effects on Serum Calcium Concentrations.

SIGNIFICANCE STATEMENT: Kidney stone disease is a common disorder with poorly understood pathophysiology. Observational and genetic studies indicate that adiposity is associated with an increased risk of kidney stone disease. However, the relative contribution of general and central adipose depots and the mechanisms by which effects of adiposity on kidney stone disease are mediated have not been defined. Using conventional and genetic epidemiological techniques, we demonstrate that general and central adiposity are independently associated with kidney stone disease. In addition, one mechanism by which central adiposity increases risk of kidney stone disease is by increasing serum calcium concentration. Therapies targeting adipose depots may affect calcium homeostasis and help to prevent kidney stone disease. BACKGROUND: Kidney stone disease affects approximately 10% of individuals in their lifetime and is frequently recurrent. The disease is linked to obesity, but the mechanisms mediating this association are uncertain. METHODS: Associations of adiposity and incident kidney stone disease were assessed in the UK Biobank over a mean of 11.6 years/person. Genome-wide association studies and Mendelian randomization (MR) analyses were undertaken in the UK Biobank, FinnGen, and in meta-analyzed cohorts to identify factors that affect kidney stone disease risk. RESULTS: Observational analyses on UK Biobank data demonstrated that increasing central and general adiposity is independently associated with incident kidney stone formation. Multivariable MR, using meta-analyzed UK Biobank and FinnGen data, established that risk of kidney stone disease increases by approximately 21% per one standard deviation increase in body mass index (BMI, a marker of general adiposity) independent of waist-to-hip ratio (WHR, a marker of central adiposity) and approximately 24% per one standard deviation increase of WHR independent of BMI. Genetic analyses indicate that higher WHR, but not higher BMI, increases risk of kidney stone disease by elevating adjusted serum calcium concentrations (ß=0.12 mmol/L); WHR mediates 12%-15% of its effect on kidney stone risk in this way. CONCLUSIONS: Our study indicates that visceral adipose depots elevate serum calcium concentrations, resulting in increased risk of kidney stone disease. These findings highlight the importance of weight loss in individuals with recurrent kidney stones and suggest that therapies targeting adipose depots may affect calcium homeostasis and contribute to prevention of kidney stone disease.