3D motion of vesicles along microtubules helps them to circumvent obstacles in cells.

Vesicle transport is regulated at multiple levels, including regulation by scaffolding proteins and the cytoskeleton. This tight regulation is essential, since slowing or stoppage of transport can cause accumulation of obstacles and has been linked to diseases. Understanding the mechanisms by which transport is regulated as well as how motor proteins overcome obstacles can give important clues as to how these mechanisms break down in disease states. Here, we describe that the cytoskeleton architecture impacts transport in a vesicle-size-dependent manner, leading to pausing of vesicles larger than the separation of the microtubules. We further develop methods capable of following 3D transport processes in living cells. Using these methods, we show that vesicles move using two different modes along the microtubule. Off-axis motion, which leads to repositioning of the vesicle in 3D along the microtubule, correlates with the presence of steric obstacles and may help in circumventing them.

Standard long IV catheters versus extended dwell catheters: A randomized comparison of ultrasound-guided catheter survival.

INTRODUCTION: Establishing peripheral intravenous (IV) access is a vital step in providing emergency care. Ten to 30% of Emergency Department (ED) patients have difficult vascular access (DVA). Even after cannulation, early failure of US-guided IV catheters is a common complication. The primary goal of this study was to compare survival of a standard long IV catheter to a longer extended dwell catheter. METHODS: This study was a prospective, randomized comparative evaluation of catheter longevity. Two catheters were used in the comparison: [1] a standard long IV catheter, the 4.78 cm 20 gauge Becton Dickinson (BD); and [2] a 6 cm 3 French (19.5 gauge) Access Scientific POWERWAND™ extended dwell catheter (EDC). Adult DVA patients in the ED with vein depths of 1.20 cm-1.60 cm and expected hospital admissions of at least 24 h were recruited. RESULTS: 120 patients were enrolled. Ultimately, 70 patients were included in the survival analysis, with 33 patients in the EDC group and 37 patients in the standard long IV group. EDC catheters had lower rates of failure (p = 0.0016). Time to median catheter survival was 4.04 days for EDC catheters versus 1.25 days for the standard long IV catheter. Multivariate survival analysis also showed a significant survival benefit for the EDC catheter (p = 0.0360). CONCLUSION: A longer extended dwell catheter represents a viable and favorable alternative to the standard longer IVs used for US-guided cannulation of veins >1.20 cm in depth. These catheters have significantly improved survival rates with similar insertion success characteristics.

Single-molecule evaluation of fluorescent protein photoactivation efficiency using an in vivo nanotemplate.

Photoswitchable fluorescent probes are central to localization-based super-resolution microscopy. Among these probes, fluorescent proteins are appealing because they are genetically encoded. Moreover, the ability to achieve a 1:1 labeling ratio between the fluorescent protein and the protein of interest makes these probes attractive for quantitative single-molecule counting. The percentage of fluorescent protein that is photoactivated into a fluorescently detectable form (i.e., the photoactivation efficiency) plays a crucial part in properly interpreting the quantitative information. It is important to characterize the photoactivation efficiency at the single-molecule level under the conditions used in super-resolution imaging. Here, we used the human glycine receptor expressed in Xenopus oocytes and stepwise photobleaching or single-molecule counting photoactivated localization microcopy (PALM) to determine the photoactivation efficiency of fluorescent proteins mEos2, mEos3.1, mEos3.2, Dendra2, mClavGR2, mMaple, PA-GFP and PA-mCherry. This analysis provides important information that must be considered when using these fluorescent proteins in quantitative super-resolution microscopy.

Acceptability of HIV testing for adolescents and young adults by delivery model: a systematic review.

HIV infections are prevalent among adolescents and young adults, of whom 44% remain unaware of their diagnosis. HIV screening presents numerous challenges including stigma, fear, and concerns about confidentiality, which may influence young people's acceptance of HIV screening and linkage to care differently from individuals in other age groups. It is imperative to understand which care delivery models are most effective in facilitating these services for youth. This systematic review analyzes the rates of HIV test acceptance and linkage to care by care delivery model for adolescents and young adults. Studies were classified into emergency department (ED), primary care/inpatient setting, community-based program, or sexually transmitted infection clinic models of care. From 6395 studies initially identified, 59 met criteria for inclusion in the final analyses. Rate of test acceptance and linkage to care were stratified by model of care delivery, gender, race, age ranges (13-17, 18-24 years) as well as site (North America vs rest of the world). A significant difference in acceptance of HIV testing was found between care models, with high rates of test acceptance in the ED setting in North America and primary care/hospital setting in the rest of the world. Similarly, linkage to care differed by model of care, with EDs having high rates of linkages to HIV care in North America. Future studies are needed to test mechanisms for optimizing outcomes for each care delivery model in addressing the unique challenges faced by adolescents and young adults.

Predictors of misperceptions, risk perceptions, and personal risk perceptions about COVID-19 by country, education and income.

Government interventions, such as mandating the use of masks and social distancing, play crucial roles in controlling the spread of pandemic infection. Adherence depends on public perceptions about pandemic risk. The goal was to explore the roles of education, income, and country on misperceptions, risk perceptions and personal risk perceptions about COVID-19. Data were extracted from 3 preregistered surveys. Binary logistic regressions were conducted to investigate the roles country, education, and income had on outcome variables. Across the USA, Canada, and UK, individuals in the highest income quartile were significantly less likely to hold misperceptions (OR=0.61, 95% CI 0.45 to 0.83) and to perceive personal risk (OR=0.38, 95% CI 0.20 to 0.75) regarding COVID-19 compared with individuals in the lowest income quartile. When comparing these income quartiles in the USA, the difference in perceived risk was heightened (OR=0.21, 95% CI 0.07 to 0.57). Citizens of the UK were more likely to have risk perceptions compared with citizens of the USA (OR=1.50, 95% CI 1.20 to 1.88). Citizens of Canada were less likely to perceive personal risk compared with US citizens (OR=0.40, 95% CI 0.23 to 0.69). Proper risk perception and understanding of COVID-19 are necessary for adherence to public health initiatives. The lowest income quartile was shown to have more misperceptions and personal risk perceptions across all 3 countries, highlighting the disproportionate impact of COVID-19 in this group. Our findings support the importance of education and income in affecting health perceptions and outcomes. Further research is needed to explore interventions to minimize misperceptions, accurately shape risk perception, and effectively communicate science.

Systematic review of cognitive behavior therapy to improve mental health of women living with HIV.

Psychological distress is highly prevalent in people living with HIV. Cognitive behavior therapy (CBT) has been associated with improved mental health outcomes in HIV-infected men who have sex with men (MSM); however, little is known of its effect in women living with HIV/AIDS (WLHA). We review current literature on CBT and its effects on depression, anxiety, stress and mental health quality of life (QOL) in WLHA. We undertook a systematic review of the literature indexed in PubMed, Medline, Psychiatry Online and ScienceDirect. Of the 273 relevant studies discovered, 158 contained duplicate data, and 105 studies did not meet the inclusion and exclusion criteria, yielding 10 studies for analysis. Data were independently extracted by each researcher, with differences resolved through discussion and consensus. For WLHA, CBT substantially improved QOL, symptoms of depression and stress, but appeared to have less impact on anxiety. Three of the six studies measuring depression outcomes showed statistically significant decreases in depression. Three of three studies measuring mental health QOL, and three of three studies measuring stress also demonstrated statistically significant improvement. Two of two studies measuring anxiety did not show statistically significant change. CBT is a promising therapy for WLHA. CBT may reduce psychological distress, improving symptoms of depression, stress and QOL. There is a need for additional, better standardized studies that examine CBT for WLHA.

Interventions to improve medication adherence in adolescents with HIV: a systematic review and meta-analysis.

As of 2017, 1.8 million people living with HIV (PLWH) were adolescents between ages 10 and 19, accounting for 5% of all PLWH and 590,000 people between the ages 15 and 24 were newly infected with HIV. Between 2004 and 2011, AIDS-related deaths increased 50% among adolescents, and optimal adolescent adherence to antiretroviral treatment (ART) is estimated at only 62% of adolescents worldwide. While there have been great strides toward achieving the UNAIDS 90-90-90 goals, adolescents remain a group lacking appropriate resources and research to achieve these. This review analyzes current interventions aimed toward increasing adolescent ART adherence. Systematic searches of EMBASE, PubMed and PsycINFO were performed using the keywords 'adolescent HIV medication adherence interventions'. The Gain Score effect size was calculated for studies reporting the Cohen's d and variance to include both prestudy and poststudy values. A random-effects model analyzed intervention significance. Authors were contacted to obtain additional data values and study clarification. Twelve studies met inclusion criteria for meta-analysis. There were no significant differences seen between control and intervention groups in medication adherence (z=-1.4714, p<0.1412), viral load (z=-0.1946, p<0.8547) or CD4+ lymphocyte count (z=0.2650, p<0.7910). There was no significant difference between studies in increasing medication adherence. Results indicate that interventions did not improve medication adherence in adolescents with HIV. However, the paucity of quantitative research available speaks to a need for more quantitative intervention studies and standardization of measures of intervention efficacy.

Comparison of Ultrasound and Plain Radiography for the Detection of Long-bone Fractures.

OBJECTIVE: To compare emergency medicine (EM) resident physicians' ability to identify long-bone fractures using ultrasound (US) versus plain radiography (X-ray). METHODS: This was an IRB-approved, randomized prospective study. Study participants included 40 EM residents at a single site. Fractures were mechanically induced in five chicken legs, and five legs were left unfractured. Chicken legs were imaged by both modalities. Participants were given 2 min to view each of the images. Participants were randomized to either US or X-ray interpretation first and randomized to viewing order within each arm. Participants documented the presence or absence of fracture and location and type of fracture when pertinent. Mean proportions and standard deviations (SDs) were analyzed using paired t-test and linear models. RESULTS: Forty residents (15 postgraduate years (PGY)-1, 12 PGY-2, 13 PGY-3) participated in the study. Thirty-one participants were male, and 19 were randomized to US first. Residents completed a mean of 185 (SD 95.8) US scans before the study in a variety of applications. Accurate fracture identification had a higher mean proportion in the US arm than the X-ray arm, 0.89 (SD 0.11) versus 0.75 (SD 0.11), respectively (P < 0.001). There was no statistically significant difference in US arm and X-ray arm for endpoints of fracture location and type. CONCLUSION: EM residents were better able to identify fractures using US compared to X-ray, especially as level of US and ED experience increased. These results encourage the use of US for the assessment of isolated extremity injury, particularly when the injury is diaphyseal.

NMDA Receptor Autoantibodies in Autoimmune Encephalitis Cause a Subunit-Specific Nanoscale Redistribution of NMDA Receptors.

Anti-N-methyl-D-aspartate receptor (NMDAR) encephalitis is a severe neuropsychiatric disorder mediated by autoantibodies against the GluN1 subunit of the NMDAR. Patients' antibodies cause cross-linking and internalization of NMDAR, but the synaptic events leading to depletion of NMDAR are poorly understood. Using super-resolution microscopy, we studied the effects of the autoantibodies on the nanoscale distribution of NMDAR in cultured neurons. Our findings show that, under control conditions, NMDARs form nanosized objects and patients' antibodies increase the clustering of synaptic and extrasynaptic receptors inside the nano-objects. This clustering is subunit specific and predominantly affects GluN2B-NMDARs. Following internalization, the remaining surface NMDARs return to control clustering levels but are preferentially retained at the synapse. Monte Carlo simulations using a model in which antibodies induce NMDAR cross-linking and disruption of interactions with other proteins recapitulated these results. Finally, activation of EphB2 receptor partially antagonized the antibody-mediated disorganization of the nanoscale surface distribution of NMDARs.

Jung and Kabbalah: imaginal and noetic aspects.

Jung made use of kabbalistic images and motifs in various parts of his opus, including in his alchemical studies, in Aion, and extensively in Mysterium Coniunctionis. He also recorded an important dream after his heart attack which made use of kabbalistic symbolism in Memories, Dreams, Reflections. In this paper I explore Jung's ideas in relation to Kabbalah, first, by differentiating between Jung's imaginal approach to kabbalistic symbolism and the noetic intention of the Kabbalah itself in its use of imaginal material. Second, I present a number of typical examples of how Jung understands (and sometimes misunderstands) kabbalistic material that he cites. Third, I briefly survey the development of the Kabbalah as an imaginal noetic system, and present a core self-understanding of kabbalists--as engaged in inner 'self-work' which intends to 'sweeten the harsh judgments of existence in their very roots'. Finally, I differentiate Jung's understanding of the psychical living symbol from the kabbalistic understanding of the mystical symbol. In this fourth section of the paper, I conclude by presenting a basic Hasidic/kabbalistic teaching on the nature and function of verbal contemplative prayer--as an illustration of the difference between the two understandings of symbolism. The four sections of the paper are framed by a 'Prelude' and a 'Coda'.

Cross-talk-free multi-color STORM imaging using a single fluorophore.

Multi-color stochastic optical reconstruction microscopy (STORM) is routinely performed; however, the various approaches for achieving multiple colors have important caveats. Color cross-talk, limited availability of spectrally distinct fluorophores with optimal brightness and duty cycle, incompatibility of imaging buffers for different fluorophores, and chromatic aberrations impact the spatial resolution and ultimately the number of colors that can be achieved. We overcome these complexities and develop a simple approach for multi-color STORM imaging using a single fluorophore and sequential labelling. In addition, we present a simple and versatile method to locate the same region of interest on different days and even on different microscopes. In combination, these approaches enable cross-talk-free multi-color imaging of sub-cellular structures.

A microfluidic platform for correlative live-cell and super-resolution microscopy.

Recently, super-resolution microscopy methods such as stochastic optical reconstruction microscopy (STORM) have enabled visualization of subcellular structures below the optical resolution limit. Due to the poor temporal resolution, however, these methods have mostly been used to image fixed cells or dynamic processes that evolve on slow time-scales. In particular, fast dynamic processes and their relationship to the underlying ultrastructure or nanoscale protein organization cannot be discerned. To overcome this limitation, we have recently developed a correlative and sequential imaging method that combines live-cell and super-resolution microscopy. This approach adds dynamic background to ultrastructural images providing a new dimension to the interpretation of super-resolution data. However, currently, it suffers from the need to carry out tedious steps of sample preparation manually. To alleviate this problem, we implemented a simple and versatile microfluidic platform that streamlines the sample preparation steps in between live-cell and super-resolution imaging. The platform is based on a microfluidic chip with parallel, miniaturized imaging chambers and an automated fluid-injection device, which delivers a precise amount of a specified reagent to the selected imaging chamber at a specific time within the experiment. We demonstrate that this system can be used for live-cell imaging, automated fixation, and immunostaining of adherent mammalian cells in situ followed by STORM imaging. We further demonstrate an application by correlating mitochondrial dynamics, morphology, and nanoscale mitochondrial protein distribution in live and super-resolution images.

Nontuberculous mycobacterial hypersensitivity pneumonitis related to a home shower: treatment and secondary prevention.

A 57-year-old physician with increasing dyspnoea and hypoxaemia had a high-resolution CT scan of the chest, which disclosed diffuse pulmonary ground glass opacities, more pronounced in the upper lobes with minimal mediastinal lymphadenopathy. Transbronchial biopsy of the right middle and lower lobes was performed, demonstrating varying degrees of well circumscribed organising granulomatous pneumonitis thought to be most consistent with hypersensitivity to nontuberculous mycobacteria. Cultures of water obtained from the patient's home shower were positive for Mycobacterium avium complex. The patient began substituting baths for showers, experiencing some gradual improvement of his symptoms. Subsequently, he installed point-of-use 0.2 micron membrane filters on his shower, and resumed regular showering after installation with continued symptomatic improvement. CT scans at 3 and 18 months revealed improvement and resolution, respectively. Four years later, he continues to shower in filtered home shower water and remains clinically well.