Direct observation of the interconversion of normal and toxic forms of α-synuclein.

Here, we use single-molecule techniques to study the aggregation of α-synuclein, the protein whose misfolding and deposition is associated with Parkinson's disease. We identify a conformational change from the initially formed oligomers to stable, more compact proteinase-K-resistant oligomers as the key step that leads ultimately to fibril formation. The oligomers formed as a result of the structural conversion generate much higher levels of oxidative stress in rat primary neurons than do the oligomers formed initially, showing that they are more damaging to cells. The structural conversion is remarkably slow, indicating a high kinetic barrier for the conversion and suggesting that there is a significant period of time for the cellular protective machinery to operate and potentially for therapeutic intervention, prior to the onset of cellular damage. In the absence of added soluble protein, the assembly process is reversed and fibrils disaggregate to form stable oligomers, hence acting as a source of cytotoxic species.

Highlighter: An optogenetic system for high-resolution gene expression control in plants.

Optogenetic actuators have revolutionized the resolution at which biological processes can be controlled. In plants, deployment of optogenetics is challenging due to the need for these light-responsive systems to function in the context of horticultural light environments. Furthermore, many available optogenetic actuators are based on plant photoreceptors that might crosstalk with endogenous signaling processes, while others depend on exogenously supplied cofactors. To overcome such challenges, we have developed Highlighter, a synthetic, light-gated gene expression system tailored for in planta function. Highlighter is based on the photoswitchable CcaS-CcaR system from cyanobacteria and is repurposed for plants as a fully genetically encoded system. Analysis of a re-engineered CcaS in Escherichia coli demonstrated green/red photoswitching with phytochromobilin, a chromophore endogenous to plants, but also revealed a blue light response likely derived from a flavin-binding LOV-like domain. We deployed Highlighter in transiently transformed Nicotiana benthamiana for optogenetic control of fluorescent protein expression. Using light to guide differential fluorescent protein expression in nuclei of neighboring cells, we demonstrate unprecedented spatiotemporal control of target gene expression. We implemented the system to demonstrate optogenetic control over plant immunity and pigment production through modulation of the spectral composition of broadband visible (white) light. Highlighter is a step forward for optogenetics in plants and a technology for high-resolution gene induction that will advance fundamental plant biology and provide new opportunities for crop improvement.

Evaluation of multi-directional speed qualities throughout adolescence in youth soccer: The non-linear nature of transfer.

Training and assessment of agility is often prioritised by soccer coaches and practitioners aiming to develop multi-directional speed. Although the importance of agility is advocated throughout childhood and adolescence, limited data evidence agility performance at different stages of adolescence. The purpose of this study was to examine differences in multi-directional speed performance in youth soccer players spanning an entire soccer academy. A total of 86 male junior-elite soccer players volunteered to participate. Anthropometric data were collected, alongside performance data from a battery of physical tests including sprinting, jumping, change of direction, reaction time, and agility. Bayesian models using log-likelihoods from posterior simulations of parameter values displayed linear or curvilinear relationships between both chronological and biological age and performance in all tests other than agility and reaction time. For agility and reaction time tests, performance improved until ~14 years of age or the estimated age of peak height velocity whereby arrested development in performance was observed. Our results demonstrate that while most performance skills improve as chronological or biological age increases, measures of agility and reaction time may not. These findings support the notion that agility performance is complex and multifaceted, eliciting unique, challenging physical demands and non-linear development.

Medical and surgical management of pneumothorax in diving and hyperbaric chambers.

The presence of a pneumothorax within a pressurized chamber represents unique diagnostic and management challenges. This is particularly the case in the medical and geographic remoteness of many chamber locations. Upon commencing chamber decompression, unvented intrapleural air expands. If its initial volume and/or degree of chamber pressure reduction is significant enough, a tension pneumothorax will result. Numerous reports chronicle failure to diagnose and manage in-chamber pneumothorax with resultant morbidity and one fatal outcome. Such cases have occurred in both medically remote and clinically based settings. This paper reviews pneumothorax and tension pneumothorax risk factors and clinical characteristics. It suggests primary medical management using the principle of oxygen-induced inherent unsaturation in concert with titrated chamber decompression designed to prevent intrapleural air expanding faster than it contracts. Should this conservative approach prove unsuccessful, and surgical venting becomes necessary or otherwise immediately indicated, interventional options are reviewed.

Pneumothorax during manned chamber operations: A summary of reported cases.

In-chamber pneumothorax has complicated medically remote professional diving operations, submarine escape training, management of decompression illness, and hospital-based provision of hyperbaric oxygen therapy. Attempts to avoid thoracotomy by combination of high oxygen partial pressure breathing (the concept of inherent unsaturation) and greatly slowed rates of chamber decompression proved successful on several occasions. When this delicate balance designed to prevent the intrapleural gas volume from expanding faster than it contracts proved futile, chest drains were inserted. The presence of pneumothorax was misdiagnosed or missed altogether with disturbing frequency, resulting in wide-ranging clinical consequences. One patient succumbed before the chamber had been fully decompressed. Another was able to ambulate unaided from the chamber before being diagnosed and managed conventionally. In between these two extremes, patients experienced varying degrees of clinical compromise, from respiratory distress to cardiopulmonary arrest, with successful resuscitation. Pneumothorax associated with manned chamber operations is commonly considered to develop while the patient is under pressure and manifests during ascent. However, published reports suggest that many were pre-existing prior to chamber entry. Risk factors included pulmonary barotrauma-induced cerebral arterial gas embolism, cardiopulmonary resuscitation, and medical or surgical procedures usually involving the lung. This latter category is of heightened importance to hyperbaric operations as an iatrogenically induced pneumothorax may take as long as 24 hours to be detected, perhaps long after a patient has been cleared for chamber exposure.

Non-pharmaceutical interventions and risk of COVID-19 infection: survey of U.K. public from November 2020 - May 2021.

INTRODUCTION: Non-pharmaceutical interventions (NPIs), such as handwashing, social distancing and face mask wearing, have been widely promoted to reduce the spread of COVID-19. This study aimed to explore the relationship between self-reported use of NPIs and COVID-19 infection. METHODS: We conducted an online questionnaire study recruiting members of the UK public from November 2020 to May 2021. The association between self-reported COVID-19 illness and reported use of NPIs was explored using logistic regression and controlling for participant characteristics, month of questionnaire completion, and vaccine status. Participants who had been exposed to COVID-19 in their household in the previous 2 weeks were excluded. RESULTS: Twenty-seven thousand seven hundred fifty-eight participants were included and 2,814 (10.1%) reported having a COVID-19 infection. The odds of COVID-19 infection were reduced with use of a face covering in unadjusted (OR 0.17 (95% CI: 0.15 to 0.20) and adjusted (aOR 0.19, 95% CI 0.16 to 0.23) analyses. Social distancing (OR 0.27, 95% CI: 0.22 to 0.31; aOR 0.35, 95% CI 0.28 to 0.43) and handwashing when arriving home (OR 0.57, 95% CI 0.46 to 0.73; aOR 0.63, 95% CI: 0.48 to 0.83) also reduced the odds of COVID-19. Being in crowded places of 10-100 people (OR 1.89, 95% CI: 1.70 to 2.11; aOR 1.62, 95% CI: 1.42 to 1.85) and > 100 people (OR 2.33, 95% CI: 2.11 to 2.58; aOR 1.73, 95% CI: 1.53 to 1.97) were both associated with increased odds of COVID-19 infection. Handwashing before eating, avoiding touching the face, and cleaning things with virus on were all associated with increased odds of COVID-19 infections. CONCLUSIONS: This large observational study found evidence for strong protective effects for individuals from use of face coverings, social distancing (including avoiding crowded places) and handwashing on arriving home on developing COVID-19 infection. We also found evidence for an increased risk associated with other behaviours, possibly from recall bias.

Exploratory assessment: Nurse-led community health worker delivered HCV intervention for people experiencing homelessness.

BACKGROUND: Getting and maintaining Hepatitis C Virus (HCV) cure is challenging among people experiencing homelessness (PEH) as a result of critical social determinants of health such as unstable housing, mental health disorders, and drug and alcohol use. OBJECTIVES: The purpose of this exploratory pilot study was to compare a registered nurse/community health worker (RN/CHW)-led HCV intervention tailored for PEH, "I am HCV Free," with a clinic-based standard of care (cbSOC) for treating HCV. Efficacy was measured by sustained virological response at 12 weeks after stopping antivirals (SVR12), and improvement in mental health, drug and alcohol use, and access to healthcare. METHODS: An exploratory randomized controlled trial design was used to assign PEH recruited from partner sites in the Skid Row Area of Los Angeles, California, to the RN/CHW or cbSOC programs. All received direct-acting antivirals. The RN/CHW group received directly observed therapy in community-based settings, incentives for taking HCV medications, and wrap-around services, including connection to additional healthcare services, housing support, and referral to other community services. For all PEH, drug and alcohol use and mental health symptoms were measured at month 2 or 3 and 5 or 6 follow-up, depending on HCV medication type, while SVR12 was measured at month 5 or 6 follow-up. RESULTS: Among PEH in the RN/CHW group, 75% (3 of 4) completed SVR12 and all three attained undetectable viral load. This was compared with 66.7% (n = 4 of 6) of the cbSOC group who completed SVR12; all four attained undetectable viral load. The RN/CHW group, as compared to the cbSOC, also showed greater improvements in mental health, and significant improvement in drug use, and access to healthcare services. DISCUSSION: While this study shows significant improvements in drug use and health service access among the RN/-CHW group, the sample size of the study limits the validity and generalizability of the results. Further studies using larger sample sizes are necessitated.

The Deceleration Deficit: A Novel Field-Based Method to Quantify Deceleration During Change of Direction Performance.

ABSTRACT: Clarke, R, Read, PJ, De Ste Croix, MBA, and Hughes, JD. The deceleration deficit: a novel field-based method to quantify deceleration during change of direction performance. J Strength Cond Res 36(9): 2434-2439, 2022-The study investigated the relationship between linear and change of direction (COD) speed performance components and the individual differences between deceleration deficit (DD) and COD deficit (CODD). Thirty-six subjects (mean ± SD : age = 20.3 ± 2.9 years; stature = 175.2 ± 7.7 cm; and body mass = 78.0 ± 16.7 kg) completed 3 trials of a 505 test in both turning directions (dominant [D]; nondominant [ND]) and 3 15-m linear sprints. Deceleration deficit was calculated by the 15-m approach in the 505 test, minus the athlete's linear 15-m sprint time. To compare individuals CODD and DD, z-scores were calculated, and moderate worthwhile changes (MWCs) were identified between these deficit z-scores. Significant correlations were identified between linear sprints and 505 time (D: r = 0.71, 0.74; P < 0.01. ND: r = 0.76, 0.75; P < 0.01) for 10-m and 15-m sprint. respectively, and between 505 performance and CODD (D: r = 0.74; P < 0.01. ND: r = 0.77; P < 0.01) and DD (D: r = 0.41, P < 0.05. ND: r = 0.44, P < 0.01). Deceleration deficit was significantly related to CODD (D: r = 0.59; P < 0.01. ND: r = 0.62; P < 0.01); however, 78% of subjects demonstrated differences between these deficit measures greater than an MWC. In conclusion, linear speed has the strongest significant relationship with 505 performance. Deceleration deficit could provide a more isolated construct than CODD which may be related to an athlete's deceleration capabilities.

Change-of-Direction Deficit vs. Deceleration Deficit: A Comparison of Limb Dominance and Inter-limb Asymmetry between Forwards and Backs in Elite Male Rugby Union Players.

The aims of the present study were to: 1) determine whether limb dominance and inter-limb asymmetry were the same across both change of direction (COD) and deceleration (DEC) deficits and, 2) determine the association between the COD and DEC-deficits and other physical performance tests in elite male rugby union players. Twenty five players performed a series of bilateral jumps, linear and COD speed tests at the end of the pre-season period. COD and DEC-deficits were calculated for both left and right sides, and inter-limb asymmetry thereafter. Kappa coefficients revealed moderate levels of agreement in limb dominance between COD and DEC-deficits (Kappa = 0.41 on left; 0.48 on right). For the direction of asymmetry, perfect levels of agreement (Kappa = 1) were evident between 505 time and COD-deficit, but only moderate levels of agreement (Kappa = 0.41) between other asymmetry measures. Pearson's r correlations showed moderate to large relationships between jumps and linear (r = -0.42 to -0.68) and COD speed (r = -0.41 to -0.58), but not with the COD-deficit (r = 0.15 to -0.31), DEC-deficit (r = 0.01 to -0.32) or asymmetry (r = 0.16 to -0.29). When analysing by playing position, backs were significantly faster than forwards over 15-m (ES = -0.86) and across all jump tests (ES = 0.86-0.94), with the exception of the squat jump. This study is the first to provide a direct comparison of the COD and DEC-deficits and highlights that limb dominance and asymmetry cannot be guaranteed between tasks.

Eye Movements in the "Morris Maze" Spatial Working Memory Task Reveal Deficits in Strategic Planning.

Analysis of eye movements can provide insights into processes underlying performance of cognitive tasks. We recorded eye movements in healthy participants and people with idiopathic Parkinson disease during a token foraging task based on the spatial working memory component of the widely used Cambridge Neuropsychological Test Automated Battery. Participants selected boxes (using a mouse click) to reveal hidden tokens. Tokens were never hidden under a box where one had been found before, such that memory had to be used to guide box selections. A key measure of performance in the task is between search errors (BSEs) in which a box where a token has been found is selected again. Eye movements were found to be most commonly directed toward the next box to be clicked on, but fixations also occurred at rates higher than expected by chance on boxes farther ahead or back along the search path. Looking ahead and looking back in this way was found to correlate negatively with BSEs and was significantly reduced in patients with Parkinson disease. Refixating boxes where tokens had already been found correlated with BSEs and the severity of Parkinson disease symptoms. It is concluded that eye movements can provide an index of cognitive planning in the task. Refixations on locations where a token has been found may also provide a sensitive indicator of visuospatial memory integrity. Eye movement measures derived from the spatial working memory task may prove useful in the assessment of executive functions as well as neurological and psychiatric diseases in the future.

Challenges threaten, opportunity awaits hyperbaric medicine and the head and neck cancer patient.

Over the past four decades, hyperbaric oxygen (HBO2) therapy has played a prominent role in both the prevention and treatment of mandibular osteoradionecrosis (ORN). It has done so on the strength of laboratory observations and clinical reports, yet only limited efficacy data. This dual role has come under increasing scrutiny in the modern radiotherapy (RT) and surgical eras. The ability to spare healthy "non-target" tissue has markedly improved since the two-dimensional planning and delivery techniques in use when HBO2's prophylactic value was first demonstrated. A recent study failed to identify this same benefit in patients who received high-precision imaging and conformal RT. HBO2 therapy is under challenge as preferred treatment for early stage ORN. A recently introduced "fibroatrophic" mechanism contrasts with the hypovascular-hypocellular-hypoxic injury pattern that formed the basis for HBO2's therapeutic use. This alternative pathophysiologic state appears to benefit from an oral antioxidant medication regimen. The continuing necessity of HBO2 in support of mandibular reconstruction for advanced ORN is in question. Microsurgery-based vascularized bone flaps increasingly represent standard care, invariably in the absence of perioperative HBO2. Renewed interest in hyperbaric oxygen as a radiation sensitizer offers some promise. Hypoxia remains a critical radio-resistant factor in many solid tumors. Malignant gliomas have been a primary focus of several small studies, with resulting improvements in local control and median survival. Hyperbaric radiation sensitization has recently addressed oropharyngeal cancer. Preliminary data indicates that addition of HBO2 to chemo-radiation standard of care is technically feasible, well tolerated and safe. A Phase II efficacy trial will investigate the potential for of HBO2 to improve progression-free and relapse-free survival in newly diagnosed locally advanced head and neck cancers. What follows is a review and summary of relevant peer-reviewed literature.

Hyperbaric medicine today: an historically noble discipline challenged by loss of critical access and overutilization - invited commentary.

As the title implies, much appears amiss with hyperbaric medicine. Long recognized for its life-saving, CNS-sparing,infection-fighting and tissue-salving attributes, its current application has been rightly called into question by a broad cross-section of health care delivery system stakeholders [1, 2, 3, 4, 5]. This paper will examine what lies behind the stunning loss of availability for a majority of the Federal Drug Administration-approved uses, arguably those for which patients have the most to gain. It will address overutilization in the context of an erosion of practice standards and widespread manipulation of the reimbursement process. It will make suggestions aimed at restoring its broader availability across the full extent of FDA-approved uses. Finally, it offers guidance to ensure that HBO2 therapy is employed only when medically necessary by adoption of the drug administration "rights" principle, namely the right indication for the right patient at the right time and only for the right amount of time.

Health care worker decompression sickness: incidence, risk and mitigation.

Inadvertent exposure to radiation, chemical agents and biological factors are well recognized hazards associated with the health care delivery system. Less well appreciated yet no less harmful is risk of decompression sickness in those who accompany patients as inside attendants (IAs) during provision of hyperbaric oxygen therapy. Unlike the above hazards where avoidance is practiced, IA exposure to decompression sickness risk is unavoidable. While overall incidence is low, when calculated as number of cases over number of exposures or potential for a case during any given exposure, employee cumulative risk, defined here as number of cases over number of IAs, or risk that an IA may suffer a case, is not. Commonly, this unique occupational environmental injury responds favorably to therapeutic recompression and a period of recuperation. There are, however, permanent and career-ending consequences, and at least two nurses have succumbed to their decompression insults. The intent of this paper is to heighten awareness of hyperbaric attendant decompression sickness. It will serve as a review of reported cases and reconcile incidence against largely ignored individual worker risk. Mitigation strategies are summarized and an approach to more precisely identify risk factors that might prompt development of consensus screening standards is proposed.

Low Stress Ion Conductance Microscopy of Sub-Cellular Stiffness.

Directly examining subcellular mechanics whilst avoiding excessive strain of a live cell requires the precise control of light stress on very small areas, which is fundamentally difficult. Here we use a glass nanopipet out of contact with the plasma membrane to both exert the stress on the cell and also accurately monitor cellular compression. This allows the mapping of cell stiffness at a lateral resolution finer than 100 nm. We calculate the stress a nanopipet exerts on a cell as the sum of the intrinsic pressure between the tip face and the plasma membrane plus its direct pressure on any glycocalyx, both evaluated from the gap size in terms of the ion current decrease. A survey of cell types confirms that an intracellular pressure of approximately 120 Pa begins to detach the plasma membrane from the cytoskeleton and reveals that the first 0.66 ± 0.09 µm of compression of a neuron cell body is much softer than previous methods have been able to detect.

Human development of the ability to learn from bad news.

Humans show a natural tendency to discount bad news while incorporating good news into beliefs (the "good news-bad news effect"), an effect that may help explain seemingly irrational risk taking. Understanding how this bias develops with age is important because adolescents are prone to engage in risky behavior; thus, educating them about danger is crucial. We reveal a striking valence-dependent asymmetry in how belief updating develops with age. In the ages tested (9-26 y), younger age was associated with inaccurate updating of beliefs in response to undesirable information regarding vulnerability. In contrast, the ability to update beliefs accurately in response to desirable information remained relatively stable with age. This asymmetry was mediated by adequate computational use of positive but not negative estimation errors to alter beliefs. The results are important for understanding how belief formation develops and might help explain why adolescents do not respond adequately to warnings.

The Potentiating Effect of an Accentuated Eccentric Load on Countermovement Jump Performance.

Hughes, JD, Massiah, RG, and Clarke, RD. The potentiating effect of an accentuated eccentric load on countermovement jump performance. J Strength Cond Res 30(12): 3450-3455, 2016-Postactivation potentiation (PAP) following a bout of high-intensity exercise of short duration is known to produce both a fatigued and a potentiated state. There has been much research in the effectiveness of various PAP protocols, yet the type of dynamic muscle action utilized has seldom been investigated. Therefore, the present study aimed to assess whether an accentuated eccentric load (AE) could enhance subsequent performance. Eleven men (21 ± 2 years, 179.1 ± 6.9 cm, 83.2 ± 10.1 kg) performed 3 countermovement jumps (CMJs) before and 6 minutes after an AE and a back squat (BSq) PAP protocol. The participant's maximum CMJ height (cm) was determined before and after both protocol. A 2-way repeated measures analysis of variance was conducted to evaluate the effect of PAP condition and time on vertical jumping performance. A significant condition × time interaction was found (p = 0.02). Post hoc tests revealed that AE PAP had a significant (p = 0.03) potentiating effect on CMJ jump height. Whereas, the BSq PAP revealed no significant differences (p = 0.32). In conclusion, this study has shown that jump performance can be enhanced by eccentric muscle action when compared with a traditional BSq PAP protocol. This may provide a more practical method for coaches to enhance short-term explosive movements in athletic populations.

Hydrodynamic trapping of molecules in lipid bilayers.

In this work we show how hydrodynamic forces can be used to locally trap molecules in a supported lipid bilayer (SLB). The method uses the hydrodynamic drag forces arising from a flow through a conical pipette with a tip radius of 1-1.5 µm, placed approximately 1 µm above the investigated SLB. This results in a localized forcefield that acts on molecules protruding from the SLB, yielding a hydrodynamic trap with a size approximately given by the size of the pipette tip. We demonstrate this concept by trapping the protein streptavidin, bound to biotin receptors in the SLB. It is also shown how static and kinetic information about the intermolecular interactions in the lipid bilayer can be obtained by relating how the magnitude of the hydrodynamic forces affects the accumulation of protein molecules in the trap.