Cross-Cultural Adaptation, Reliability, and Convergent Validity of Occupational Self-Assessment for Brazilian Portuguese.

The Occupational Self-Assessment version 2.2 (OSA) is a self-assessment of the client's occupational competence and values. To describe the process of cross-cultural adaptation of OSA into Brazilian Portuguese (OSA-Brazil) and examine its reliability and validity for use with the Brazilian population. Assessment translation was guided by two international guidelines for cross-cultural adaptation of standardized instruments. Face validity was tested with 24 participants. Internal consistency, test-retest reliability, and convergent validity were tested with a convenience sample of 40 participants. The cross-cultural adaptation process concluded with a consensus among the expert panel review (r > 80%) and evidence of strong face validity. The OSA-Brazil demonstrated appropriate test-retest reliability (r > 0.70) and convergent validity with the 36-Item Short-Form Health Survey (SF-36) (p < .05). The OSA-Brazil has good face validity, test-retest reliability, and convergent validity. The assessment can be used by the Brazilian occupational therapists to assess client's occupational competence.

Particle size influences decay rates of environmental DNA in aquatic systems.

Environmental DNA (eDNA) analysis is a powerful tool for remote detection of target organisms. However, obtaining quantitative and longitudinal information from eDNA data is challenging, requiring a deep understanding of eDNA ecology. Notably, if the various size components of eDNA decay at different rates, and we can separate them within a sample, their changing proportions could be used to obtain longitudinal dynamics information on targets. To test this possibility, we conducted an aquatic mesocosm experiment in which we separated fish-derived eDNA components using sequential filtration to evaluate the decay rate and changing proportion of various eDNA particle sizes over time. We then fit four alternative mathematical decay models to the data, building towards a predictive framework to interpret eDNA data from various particle sizes. We found that medium-sized particles (1-10 µm) decayed more slowly than other size classes (i.e., <1 and > 10 µm), and thus made up an increasing proportion of eDNA particles over time. We also observed distinct eDNA particle size distribution (PSD) between our Common carp and Rainbow trout samples, suggesting that target-specific assays are required to determine starting eDNA PSDs. Additionally, we found evidence that different sizes of eDNA particles do not decay independently, with particle size conversion replenishing smaller particles over time. Nonetheless, a parsimonious mathematical model where particle sizes decay independently best explained the data. Given these results, we suggest a framework to discern target distance and abundance with eDNA data by applying sequential filtration, which theoretically has both metabarcoding and single-target applications.

Antibiotic release from PMMA spacers and PMMA beads measured with ELISA: Assessment of in vitro samples and drain fluid samples of patients.

For prosthetic joint infections, antibiotic loaded poly methyl methacrylate (PMMA) spacer or beads can be used to release high concentrations of antibiotics locally at the infection site, while minimizing systemic toxicity. The aim of this study is to determine in vitro and in vivo pharmacokinetic release profile of antibiotics from PMMA spacers and PMMA beads. For the in vitro experiment, the PMMA spacers or beads were submerged in phosphate-buffered saline and gentamicin concentrations were determined from collected specimen at several times points, measured with enzyme-linked immunosorbent assays (ELISA). To assess the in vivo antibiotic release profile of different spacers, wound drainage fluid samples were collected after implantation of a spacer over a period of maximum 14 days. After 48 h, the burst gentamicin concentration elution was 9862 ± 1782 ng/ml (mean ± SD) from spacers versus 38,394 ± 7071 ng/ml (mean ± SD) for beads. Over 35 days, spacers had eluted a cumulative mean concentration of 13,812 ± 3548 versus 55,048 ± 12,006 ng/ml for beads (p < 0.001). Clinical samples of patients with a Vancogenx® spacer showed higher gentamicin release than Refobacin™ spacers (p < 0.001). This is the first study that measured the release data of local antibiotics with ELISA. Compare to spacers, the exact release values of gentamicin from PMMA beads are more than 10 times higher and reached a maximum much later than spacers. This makes the use of PMMA beads more preferable to use for treatment of the infection itself.

Changes in the rest-activity rhythm in migraine patients are associated with anxiety symptoms

Objective: To characterize rest-activity rhythm in chronic migraine (CM) and to investigate the relationship between this rhythm and depressive and anxiety symptoms in patients with CM. Methods: This was a study of adults aged 20 to 40 years. The rest-activity rhythm of patients with CM (n=23) and non-headache controls (NH, n=23) was assessed by actigraphy for 15 days, and they completed the following assessments: Visual Analogue Scale for pain intensity; Headache Diary; Headache Impact Test-6; Morningness-Eveningness Questionnaire; Pittsburgh Sleep Quality Index; Epworth Sleepiness Scale; Beck Depression Inventory; and State-Trait Anxiety Inventory. Results: Patients with CM showed less activity over 24 hours and more fragmented sleep. Reduced interdaily stability of the rest-activity rhythm was observed, with less robustness of this rhythm in the CM group. Multiple linear regressions revealed a significant association between the rest-activity rhythm and trait anxiety variables in patients with CM, specifically regarding the relative amplitude of the cycle, activity throughout 24 hours and during sleep, and robustness of the rest-activity rhythm. Conclusions: Our findings provide evidence that the robustness of the rest-activity rhythm, activity throughout 24 hours, and sleep fragmentation are associated with trait anxiety in patients with CM. Clinical trial registration: Brazilian Clinical Trials Registry (registration number: RBR-4M5J4S).

Bilateral osteochondritis dissecans of the shoulder.

Osteochondritis dissecans as a pathology is pre-dominantly described in the knee, elbow and ankle. Osteochondritis dissecans of the humeral head is a more uncommon reported injury. We present a case of a bilateral osteochondritis dissecans of the humeral head in a 16-year-old soccer player and an algorithm for treatment of OCD of the humeral head. To our knowledge this has never been described so specifically in literature before.

Changes in the rest-activity rhythm in migraine patients are associated with anxiety symptoms.

OBJECTIVE: To characterize rest-activity rhythm in chronic migraine (CM) and to investigate the relationship between this rhythm and depressive and anxiety symptoms in patients with CM. METHODS: This was a study of adults aged 20 to 40 years. The rest-activity rhythm of patients with CM (n=23) and non-headache controls (NH, n=23) was assessed by actigraphy for 15 days, and they completed the following assessments: Visual Analogue Scale for pain intensity; Headache Diary; Headache Impact Test-6; Morningness-Eveningness Questionnaire; Pittsburgh Sleep Quality Index; Epworth Sleepiness Scale; Beck Depression Inventory; and State-Trait Anxiety Inventory. RESULTS: Patients with CM showed less activity over 24 hours and more fragmented sleep. Reduced interdaily stability of the rest-activity rhythm was observed, with less robustness of this rhythm in the CM group. Multiple linear regressions revealed a significant association between the rest-activity rhythm and trait anxiety variables in patients with CM, specifically regarding the relative amplitude of the cycle, activity throughout 24 hours and during sleep, and robustness of the rest-activity rhythm. CONCLUSIONS: Our findings provide evidence that the robustness of the rest-activity rhythm, activity throughout 24 hours, and sleep fragmentation are associated with trait anxiety in patients with CM.

Inhibitors of heat shock protein 70 (Hsp70) with enhanced metabolic stability reduce tau levels.

The molecular chaperone, Heat Shock Protein 70 (Hsp70), is an emerging drug target for neurodegenerative diseases, because of its ability to promote degradation of microtubule-associated protein tau (MAPT/tau). Recently, we reported YM-08 as a brain penetrant, allosteric Hsp70 inhibitor, which reduces tau levels. However, the benzothiazole moiety of YM-08 is vulnerable to metabolism by CYP3A4, limiting its further application as a chemical probe. In this manuscript, we designed and synthesized seventeen YM-08 derivatives by systematically introducing halogen atoms to the benzothiazole ring and shifting the position of the heteroatom in a distal pyridine. In microsome assays, we found that compound JG-23 has 12-fold better metabolic stability and it retained the ability to reduce tau levels in two cell-based models. These chemical probes of Hsp70 are expected to be useful tools for studying tau homeostasis.

A Phosphoramidate Strategy Enables Membrane Permeability of a Non-nucleotide Inhibitor of the Prolyl Isomerase Pin1.

The membrane permeability of nucleotide-based drugs, such as sofosbuvir (Sovaldi), requires installation of phosphate-caging groups. One strategy, termed "ProTide", masks the anionic phosphate through an N-linked amino ester and an O-linked aromatic phospho-ester, such that release of the active drug requires consecutive enzymatic liberation by an esterase and then a phosphoramidase, such as Hint1. Because Hint1 is known to be selective for nucleotides, it was not clear if the ProTide approach could be deployed for non-nucleotides. Here, we demonstrate that caging of a phosphate-containing inhibitor of the prolyl isomerase Pin1 increases its permeability. Moreover, this compound was processed by both esterase and phosphoramidase activity, releasing the active molecule to bind and inhibit Pin1 in cells. Thus, Hint1 appears to recognize a broader set of substrates than previously appreciated. It seems possible that other potent, but impermeable, phosphate-containing inhibitors might likewise benefit from this approach.

Robust Sequence Determinants of α-Synuclein Toxicity in Yeast Implicate Membrane Binding.

Protein conformations are shaped by cellular environments, but how environmental changes alter the conformational landscapes of specific proteins in vivo remains largely uncharacterized, in part due to the challenge of probing protein structures in living cells. Here, we use deep mutational scanning to investigate how a toxic conformation of α-synuclein, a dynamic protein linked to Parkinson's disease, responds to perturbations of cellular proteostasis. In the context of a course for graduate students in the UCSF Integrative Program in Quantitative Biology, we screened a comprehensive library of α-synuclein missense mutants in yeast cells treated with a variety of small molecules that perturb cellular processes linked to α-synuclein biology and pathobiology. We found that the conformation of α-synuclein previously shown to drive yeast toxicity-an extended, membrane-bound helix-is largely unaffected by these chemical perturbations, underscoring the importance of this conformational state as a driver of cellular toxicity. On the other hand, the chemical perturbations have a significant effect on the ability of mutations to suppress α-synuclein toxicity. Moreover, we find that sequence determinants of α-synuclein toxicity are well described by a simple structural model of the membrane-bound helix. This model predicts that α-synuclein penetrates the membrane to constant depth across its length but that membrane affinity decreases toward the C terminus, which is consistent with orthogonal biophysical measurements. Finally, we discuss how parallelized chemical genetics experiments can provide a robust framework for inquiry-based graduate coursework.

Cross-Cultural Adaptation and Face Validity of the Functional Mobility Assessment into Brazilian Portuguese.

Introduction. The Functional Mobility Assessment (FMA) measures satisfaction with mobility devices in daily life. However, in Brazil, there is a lack of instruments which measure functional mobility. OBJECTIVE: We aim to report the cross-cultural adaptation process and face validity of the FMA for use in Brazil. METHODS: Two international guidelines were used in the cross-cultural adaptation process. Two independent translators translated the instrument from English to Brazilian Portuguese, and the two versions were reconciled. Two different translators back translated this reconciled version, and an expert committee analysed the resulting synthesis. For face validity, the FMA was applied with 24 participants, divided into two groups, users with disabilities (n = 12) and occupational therapy students (n = 12) and occupational therapy students (. RESULTS: The cross-cultural adaptation of the FMA was concluded, and its face validity presented that both groups understood most or completely all instrument items. CONCLUSION: The Brazilian version of FMA is now available in Brazilian Portuguese and has face validation. Further studies should test its psychometric properties.

Author Correction: Hsp70 and Hsp40 inhibit an inter-domain interaction necessary for transcriptional activity in the androgen receptor.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Genome skimming is a low-cost and robust strategy to assemble complete mitochondrial genomes from ethanol preserved specimens in biodiversity studies.

Global loss of biodiversity is an ongoing process that concerns both local and global authorities. Studies of biodiversity mainly involve traditional methods using morphological characters and molecular protocols. However, conventional methods are a time consuming and resource demanding task. The development of high-throughput sequencing (HTS) techniques has reshaped the way we explore biodiversity and opened a path to new questions and novel empirical approaches. With the emergence of HTS, sequencing the complete mitochondrial genome became more accessible, and the number of genome sequences published has increased exponentially during the last decades. Despite the current state of knowledge about the potential of mitogenomics in phylogenetics, this is still a relatively under-explored area for a multitude of taxonomic groups, especially for those without commercial relevance, non-models organisms and with preserved DNA. Here we take the first step to assemble and annotate the genomes from HTS data using a new protocol of genome skimming which will offer an opportunity to extend the field of mitogenomics to under-studied organisms. We extracted genomic DNA from specimens preserved in ethanol. We used Nextera XT DNA to prepare indexed paired-end libraries since it is a powerful tool for working with diverse samples, requiring a low amount of input DNA. We sequenced the samples in two different Illumina platform (MiSeq or NextSeq 550). We trimmed raw reads, filtered and had their quality tested accordingly. We performed the assembly using a baiting and iterative mapping strategy, and the annotated the putative mitochondrion through a semi-automatic procedure. We applied the contiguity index to access the completeness of each new mitogenome. Our results reveal the efficiency of the proposed method to recover the whole mitogenomes of preserved DNA from non-model organisms even if there are gene rearrangement in the specimens. Our findings suggest the potential of combining the adequate platform and library to the genome skimming as an innovative approach, which opens a new range of possibilities of its use to obtain molecular data from organisms with different levels of preservation.

Temperature-jump solution X-ray scattering reveals distinct motions in a dynamic enzyme.

Correlated motions of proteins are critical to function, but these features are difficult to resolve using traditional structure determination techniques. Time-resolved X-ray methods hold promise for addressing this challenge, but have relied on the exploitation of exotic protein photoactivity, and are therefore not generalizable. Temperature jumps, through thermal excitation of the solvent, have been utilized to study protein dynamics using spectroscopic techniques, but their implementation in X-ray scattering experiments has been limited. Here, we perform temperature-jump small- and wide-angle X-ray scattering measurements on a dynamic enzyme, cyclophilin A, demonstrating that these experiments are able to capture functional intramolecular protein dynamics on the microsecond timescale. We show that cyclophilin A displays rich dynamics following a temperature jump, and use the resulting time-resolved signal to assess the kinetics of conformational changes. Two relaxation processes are resolved: a fast process is related to surface loop motions, and a slower process is related to motions in the core of the protein that are critical for catalytic turnover.

Hsp70 and Hsp40 inhibit an inter-domain interaction necessary for transcriptional activity in the androgen receptor.

Molecular chaperones such as Hsp40 and Hsp70 hold the androgen receptor (AR) in an inactive conformation. They are released in the presence of androgens, enabling transactivation and causing the receptor to become aggregation-prone. Here we show that these molecular chaperones recognize a region of the AR N-terminal domain (NTD), including a FQNLF motif, that interacts with the AR ligand-binding domain (LBD) upon activation. This suggests that competition between molecular chaperones and the LBD for the FQNLF motif regulates AR activation. We also show that, while the free NTD oligomerizes, binding to Hsp70 increases its solubility. Stabilizing the NTD-Hsp70 interaction with small molecules reduces AR aggregation and promotes its degradation in cellular and mouse models of the neuromuscular disorder spinal bulbar muscular atrophy. These results help resolve the mechanisms by which molecular chaperones regulate the balance between AR aggregation, activation and quality control.

A prediction model of surgical site infection after instrumented thoracolumbar spine surgery in adults.

PURPOSE: The aim of this study was to develop and internally validate a multivariable model for accurate prediction of surgical site infection (SSI) after instrumented spine surgery using a large cohort of a Western European academic center. METHOD: Data of potential predictor variables were collected in 898 adult patients who underwent instrumented posterior fusion of the thoracolumbar spine. We used logistic regression analysis to develop the prediction model for SSI. The ability to discriminate between those who developed SSI and those who did not was quantified as the area under the receiver operating characteristic curve (AUC). Model calibration was evaluated by visual inspection of the calibration plot and by computing the Hosmer and Lemeshow goodness-of-fit test. RESULTS: Sixty patients (6.7%) were diagnosed with an SSI. After backward stepwise elimination of predictor variables, we formulated a model in which an individual's risk of an SSI can be computed. Age, body mass index, ASA score, degenerative or revision surgery and NSAID use appeared to be independent predictor variables for the risk of SSI. The AUC was 0.72 (95% CI 0.65-0.79), indicating reasonable discriminative ability. CONCLUSIONS: We developed and internally validated a prediction model for SSI after instrumented thoracolumbar spine surgery using predictor variables of standard clinical practice that showed reasonable discriminative ability and calibration. Identification of patients at risk for SSI allows for individualized patient risk assessment with better patient-specific counseling and may accelerate the implementation of multi-disciplinary strategies for reduction of SSI. These slides can be retrieved under Electronic Supplementary Material.

Extending chemical perturbations of the ubiquitin fitness landscape in a classroom setting reveals new constraints on sequence tolerance.

Although the primary protein sequence of ubiquitin (Ub) is extremely stable over evolutionary time, it is highly tolerant to mutation during selection experiments performed in the laboratory. We have proposed that this discrepancy results from the difference between fitness under laboratory culture conditions and the selective pressures in changing environments over evolutionary timescales. Building on our previous work (Mavor et al., 2016), we used deep mutational scanning to determine how twelve new chemicals (3-Amino-1,2,4-triazole, 5-fluorocytosine, Amphotericin B, CaCl2, Cerulenin, Cobalt Acetate, Menadione, Nickel Chloride, p-Fluorophenylalanine, Rapamycin, Tamoxifen, and Tunicamycin) reveal novel mutational sensitivities of ubiquitin residues. Collectively, our experiments have identified eight new sensitizing conditions for Lys63 and uncovered a sensitizing condition for every position in Ub except Ser57 and Gln62. By determining the ubiquitin fitness landscape under different chemical constraints, our work helps to resolve the inconsistencies between deep mutational scanning experiments and sequence conservation over evolutionary timescales.

A Retrospective Analysis of Deep Surgical Site Infection Treatment after Instrumented Spinal Fusion with the Use of Supplementary Local Antibiotic Carriers.

Background: There is no generally established treatment algorithm for the management of surgical site infection (SSI) and non-union after instrumented spinal surgery. In contrast to infected hip- and knee- arthroplasties, the use of a local gentamicin impregnated carrier in spinal surgery has not been widely reported in literature. Patients and methods: We studied 48 deep SSI and non-union patients after instrumented spine surgery, treated between 1999 and 2016. The minimum follow-up was 1.5 years. All infections were treated with a treatment-regimen consisting of systemic antibiotics and repetitive surgical debridement, supplemented with local gentamicin releasing carriers. We analysed the outcome of this treatment regimen with regard to healing of the infection, as well as patient- and surgery-characteristics of failed and successfully treated patients. Results: 42 of the 48 (87.5%) patients showed successful resolution of the SSI without recurrence with a stable spine at the end of treatment. 36 patients' SSI were treated with debridement, local antibiotics, and retention or eventual restabilization of the instrumentation in case of loosening. 3 patients were treated without local antibiotics because of very mild infection signs during the revision operation. 3 patients were treated with debridement, local antibiotics and removal of instrumentation. One of these patients was restabilized in a second procedure. Infection persisted or recurred in 6 patients. These patients had a worse physical status with a higher ASA-score. Staphylococcus aureus was the most frequent causative microorganism. Interpretation: Debridement and retention of the instrumentation, in combination with systemic antibiotics and the addition of local antibiotics provided a successful treatment for SSI and non-union after instrumented spinal fusion.

External validation of a prediction model for surgical site infection after thoracolumbar spine surgery in a Western European cohort.

BACKGROUND: A prediction model for surgical site infection (SSI) after spine surgery was developed in 2014 by Lee et al. This model was developed to compute an individual estimate of the probability of SSI after spine surgery based on the patient's comorbidity profile and invasiveness of surgery. Before any prediction model can be validly implemented in daily medical practice, it should be externally validated to assess how the prediction model performs in patients sampled independently from the derivation cohort. METHODS: We included 898 consecutive patients who underwent instrumented thoracolumbar spine surgery. To quantify overall performance using Nagelkerke's R2 statistic, the discriminative ability was quantified as the area under the receiver operating characteristic curve (AUC). We computed the calibration slope of the calibration plot, to judge prediction accuracy. RESULTS: Sixty patients developed an SSI. The overall performance of the prediction model in our population was poor: Nagelkerke's R2 was 0.01. The AUC was 0.61 (95% confidence interval (CI) 0.54-0.68). The estimated slope of the calibration plot was 0.52. CONCLUSIONS: The previously published prediction model showed poor performance in our academic external validation cohort. To predict SSI after instrumented thoracolumbar spine surgery for the present population, a better fitting prediction model should be developed.

Frontline Science: High fat diet and leptin promote tumor progression by inducing myeloid-derived suppressor cells.

Obesity is a risk factor for cancer incidence and cancer mortality. The association of obesity and cancer is attributed to multiple factors, but the tightest linkage is with the chronic, low-grade inflammation that accompanies obesity. Myeloid-derived suppressor cells (MDSC) are known facilitators of cancer progression that act by suppressing the activation and function of tumor-reactive T cells. Because MDSC quantity and function are driven by chronic inflammation, we hypothesized that MDSC may accumulate in obese individuals and facilitate tumor growth by suppressing antitumor immunity. To test this hypothesis, tumor-bearing mice on a high fat or low fat diet (HFD or LFD) were assessed for tumor progression and the metabolic dysfunction associated with obesity. HFD enhanced the accumulation of MDSC, and the resulting MDSC had both beneficial and detrimental effects. HFD-induced MDSC protected mice against diet-induced metabolic dysfunction and reduced HFD-associated inflammation, but also increased the accumulation of fat, enhanced tumor progression, and spontaneous metastasis and reduced survival time. HFD-induced MDSC facilitated tumor growth by limiting the activation of tumor-reactive CD8+ T cells. Leptin, an adipokine that regulates appetite satiety and is overexpressed in obesity, undergoes crosstalk with MDSC in which leptin drives the accumulation of MDSC while MDSC down-regulate the production of leptin. Collectively, these studies demonstrate that although MDSC protect against some metabolic dysfunction associated with HFD they enhance tumor growth in HFD mice and that leptin is a key regulator linking HFD, chronic inflammation, immune suppression, and tumor progression.