Movement preferences of the wrist and forearm during activities of daily living.

BACKGROUND: During activities of daily living, the main degrees of freedom of the forearm and wrist-forearm pronation-supination (PS), wrist flexion-extension (FE), and wrist radial-ulnar deviation (RUD)-combine seamlessly to allow the hand to engage with and manipulate objects in our environment. Yet the combined behavior of these three degrees of freedom is relatively unknown. PURPOSE: To provide a characterization of natural forearm and wrist kinematics (joint configuration, movement direction, and speed) during activities of daily living. STUDY DESIGN: This is a descriptive cross-sectional study. METHODS: Ten healthy subjects performed 24 activities of daily living chosen to represent a wide variety of activities, while we measured their PS, FE, and RUD angles using electromagnetic motion capture. The orientation of the forearm and wrist was represented in the three-dimensional "configuration space" spanned by PS, FE, and RUD. From the time course of forearm and wrist orientation in configuration space, we extracted three-dimensional distributions of joint configuration, movement direction, and speed. RESULTS: Most joint configurations were focused in a relatively small area: subjects spent roughly 50% of the time in the central 20% of their functional range of motion. Some movement directions were significantly more common than others (p < 0.001); in particular, the direction of the dart-thrower's motion (DTM) was about three times more common than motion perpendicular to it. Most movements were slow: the likelihood of moving at increasing speeds dropped off exponentially. Interestingly, the most common high-speed motion combined the DTM with a twist from pronation to supination. As this motion allows one to pick up an object in front of one's body and bring it to the head, it is essential for self-care. Thus, although many activities of daily living follow the DTM without significant forearm rotation, the greatest importance of the DTM may lie in its combination with forearm rotation. CONCLUSIONS: Despite the wide variety of activities, we found evidence of preferred movement behavior, and this behavior showed significant coupling between the wrist and forearm.

Rapid Identification of Drug Resistance and Phylogeny in M. tuberculosis, Directly from Sputum Samples.

Tuberculosis (TB) remains one of the most important infectious diseases globally. Establishing a resistance profile from the initial TB diagnosis is a priority. Rapid molecular tests evaluate only the most common genetic variants responsible for resistance to certain drugs, and Whole Genome Sequencing (WGS) needs culture prior to next-generation sequencing (NGS), limiting their clinical value. Targeted sequencing (TS) from clinical samples avoids these drawbacks, providing a signature of genetic markers that can be associated with drug resistance and phylogeny. In this study, a proof-of-concept protocol was developed for detecting genomic variants associated with drug resistance and for the phylogenetic classification of Mycobacterium Tuberculosis (Mtb) in sputum samples. Initially, a set of Mtb reference strains from the WHO were sequenced (WGS and TS). The results from the protocol agreed >95% with WHO reported data and phenotypic drug susceptibility testing (pDST). Lineage genetics results were 100% concordant with those derived from WGS. After that, the TS protocol was applied to sputum samples from TB patients to detect resistance to first- and second-line drugs and derive phylogeny. The accuracy was >90% for all evaluated drugs, except Eto/Pto (77.8%), and 100% were phylogenetically classified. The results indicate that the described protocol, which affords the complete drug resistance profile and phylogeny of Mtb from sputum, could be useful in the clinical area, advancing toward more personalized and more effective treatments in the near future. IMPORTANCE The COVID-19 pandemic negatively affected the progress in accessing essential Tuberculosis (TB) services and reducing the burden of TB disease, resulting in a decreased detection of new cases and increased deaths. Generating molecular diagnostic tests with faster results without losing reliability is considered a priority. Specifically, developing an antimicrobial resistance profile from the initial stages of TB diagnosis is essential to ensure appropriate treatment. Currently available rapid molecular tests evaluate only the most common genetic variants responsible for resistance to certain drugs, limiting their clinical value. In this work, targeted sequencing on sputum samples from TB patients was used to identify Mycobacterium tuberculosis mutations in genes associated with drug resistance and to derive a phylogeny of the infecting strain. This protocol constitutes a proof-of-concept toward the goal of helping clinicians select a timely and appropriate treatment by providing them with actionable information beyond current molecular approaches.

A bioinformatics pipeline for Mycobacterium tuberculosis sequencing that cleans contaminant reads from sputum samples.

Next-Generation Sequencing (NGS) is widely used to investigate genomic variation. In several studies, the genetic variation of Mycobacterium tuberculosis has been analyzed in sputum samples without previous culture, using target enrichment methodologies for NGS. Alignments obtained by different programs generally map the sequences under default parameters, and from these results, it is assumed that only Mycobacterium reads will be obtained. However, variants of interest microorganism in clinical samples can be confused with a vast collection of reads from other bacteria, viruses, and human DNA. Currently, there are no standardized pipelines, and the cleaning success is never verified since there is a lack of rigorous controls to identify and remove reads from other sputum-microorganisms genetically similar to M. tuberculosis. Therefore, we designed a bioinformatic pipeline to process NGS data from sputum samples, including several filters and quality control points to identify and eliminate non-M. tuberculosis reads to obtain a reliable genetic variant report. Our proposal uses the SURPI software as a taxonomic classifier to filter input sequences and perform a mapping that provides the highest percentage of Mycobacterium reads, minimizing the reads from other microorganisms. We then use the filtered sequences to perform variant calling with the GATK software, ensuring the mapping quality, realignment, recalibration, hard-filtering, and post-filter to increase the reliability of the reported variants. Using default mapping parameters, we identified reads of contaminant bacteria, such as Streptococcus, Rhotia, Actinomyces, and Veillonella. Our final mapping strategy allowed a sequence identity of 97.8% between the input reads and the whole M. tuberculosis reference genome H37Rv using a genomic edit distance of three, thus removing 98.8% of the off-target sequences with a Mycobacterium reads loss of 1.7%. Finally, more than 200 unreliable genetic variants were removed during the variant calling, increasing the report's reliability.

Cyclic Peptide [R4W4] in Improving the Ability of First-Line Antibiotics to Inhibit Mycobacterium tuberculosis Inside in vitro Human Granulomas.

Tuberculosis (TB) is currently one of the leading causes of global mortality. Medical non-compliance due to the length of the treatment and antibiotic side effects has led to the emergence of multidrug-resistant (MDR) strains of Mycobacterium tuberculosis (M. tb) that are difficult to treat. A current therapeutic strategy attempting to circumvent this issue aims to enhance drug delivery to reduce the duration of the antibiotic regimen or dosage of first-line antibiotics. One such agent that may help is cyclic peptide [R4W4], as it has been shown to have antibacterial properties (in combination with tetracycline) against methicillin-resistant Staphylococcus aureus (MRSA) in the past. The objective of this study is to test cyclic peptide [R4W4] both alone and in combination with current first-line antibiotics (either isoniazid or pyrazinamide) to study the effects of inhibition of M. tb inside in vitro human granulomas. Results from our studies indicate that [R4W4] is efficacious in controlling M. tb infection in the granulomas and has enhanced inhibitory effects in the presence of first-line antibiotics.

Antimycobacterial Effects of Everolimus in a Human Granuloma Model.

Mycobacterium tuberculosis (M. tb) has been historically and is currently a threat to global public health. First-line antibiotics have been effective but proven to be burdensome as they have many potential adverse side effects. There has been a recent increase in the number of active tuberculosis (TB) cases due to a prevalence of multidrug and extensively drug-resistant strains of M. tb, and an increasing number of highly susceptible people such as those with Type 2 Diabetes (T2DM) and human immunodeficiency virus (HIV) infection. Multidrug-resistant M. tb infection (MDR-TB) is challenging to treat with existing therapeutics, so novel therapeutics and treatment strategies must be developed. Host-Directed Therapy (HDT) has been a potential target mechanism for effective clearance of infection. Host cell autophagy plays an essential role in antibacterial defense. The mammalian target of rapamycin (mTOR) has been negatively correlated with autophagy induction. Everolimus is an mTOR inhibitor that induces autophagy, but with higher water solubility. Therefore, targeting the mTOR pathway has the potential to develop novel and more effective combination drug therapy for TB. This study tested the effect of everolimus, alone and in combination with current first-line antibiotics (isoniazid and pyrazinamide), on the inhibition of M. tb inside in vitro human granulomas. We found that M. tb-infected in vitro granulomas treated with everolimus alone resulted in significantly decreased M. tb burden compared to similar granulomas in the control group. Cells treated with everolimus doses of either 1 nM or 2 nM in conjunction with pyrazinamide (PZA) produced a significant reduction in intracellular M. tb burden. Treatment groups that received everolimus alone in either 1 nM or 2 nM doses experienced a significant reduction in oxidative stress. Additionally, samples treated with 2 nM everolimus alone were observed to have significantly higher levels of autophagy and mTOR inhibition as well. Results from this study indicate that everolimus is efficacious in controlling M. tb infection in the granulomas and has additive effects when combined with the anti-TB drugs, isoniazid and pyrazinamide. This study has shown that everolimus is a promising host-directed therapeutic in the context of in vitro granuloma M. tb infection. Further study is warranted to better characterize these effects.

Elucidating the Efficacy of the Bacille Calmette-Guérin Vaccination in Conjunction with First Line Antibiotics and Liposomal Glutathione.

Mycobacterium tuberculosis (M. tb) is the etiological agent that is responsible for causing tuberculosis (TB). Although every year M. tb infection affects millions of people worldwide, the only vaccine that is currently available is the Bacille Calmette-Guérin (BCG) vaccine. However, the BCG vaccine has varying efficacy. Additionally, the first line antibiotics administered to patients with active TB often cause severe complications and side effects. To improve upon the host response mechanism in containing M. tb infection, our lab has previously shown that the addition of the biological antioxidant glutathione (GSH) has profound antimycobacterial effects. The aim of this study is to understand the additive effects of BCG vaccination and ex-vivo GSH enhancement in improving the immune responses against M. tb in both groups; specifically, their ability to mount an effective immune response against M. tb infection, maintain CD4+ and CD8+ T cells in the granulomas, their response to liposomal glutathione (L-GSH), with varying suboptimal levels of the first line antibiotics isoniazid (INH) and pyrazinamide (PZA), the expressions of programmed death receptor 1 (PD-1), and their ability to induce autophagy. Our results revealed that BCG vaccination, along with GSH enhancement, can prevent the loss of CD4+ and CD8+ T cells in the granulomas and improve the control of M. tb infection by decreasing the expressions of PD-1 and increasing autophagy and production of the cytokines interferon gamma IFN-γ and tumor necrosis factor-α (TNF-α).

Dietary Caffeine and Polyphenol Supplementation Enhances Overall Metabolic Rate and Lipid Oxidation at Rest and After a Bout of Sprint Interval Exercise.

Jo, E, Lewis, KL, Higuera, D, Hernandez, J, Osmond, AD, Directo, DJ, and Wong, M. Dietary caffeine and polyphenol supplementation enhances overall metabolic rate and lipid oxidation at rest and after a bout of sprint interval exercise. J Strength Cond Res 30(7): 1871-1879, 2016-The purpose of this study was to investigate the effects of a caffeine-polyphenolic supplement on (a) metabolic rate and fat oxidation at rest and after a bout of sprint interval exercise (SIE) and (b) SIE performance. In a double-blind, randomized, placebo-controlled, crossover study and after an initial familiarization visit, 12 subjects (male: n = 11; female: n = 1) (body mass = 76.1 ± 2.2 kg; height = 169.8 ± 1.6 cm; body mass index = 22.7 ± 3.0 kg·m; body fat % = 21.6 ± 2.0%) underwent 2 testing sessions during which time they consumed either a caffeine-polyphenol supplement or placebo. After supplementation, resting energy expenditure, heart rate (HR), and blood pressure (BP) were assessed. Subsequently, subjects performed 30 minutes of SIE while researchers collected performance data. Subjects were then tested for post-SIE energy expenditure, HR, and BP. The caffeine-polyphenol treatment resulted in significantly (p ≤ 0.05) greater energy expenditure (+7.99% rest; +10.16% post-SIE), V[Combining Dot Above]O2 (+9.64% rest; +12.10% post-SIE), and fat oxidation rate (+10.60% rest; +9.76% post-SIE) vs. placebo at rest and post-SIE. No significant differences were detected for peak and average power at all sprint intervals between treatments. Post-SIE HR was significantly (p ≤ 0.05) greater with caffeine-polyphenol supplementation vs. placebo (90.8 ± 3.5 vs. 85.1 ± 3.6 b·min). There were no significant between-treatment differences for BP. It may be concluded that the observed thermogenic response after SIE was directly attributable to caffeine-polyphenol supplementation as opposed to an indirect manifestation of enhanced performance and work output. Collectively, these results corroborate the use of dietary caffeine and polyphenols to support efforts to reduce adiposity and improve overall body composition especially in conjunction with SIE.