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Abstract The Tri-Service Microbiome Consortium (TSMC) was founded to enhance collaboration, coordination, and communication of microbiome research among DoD organizations and to facilitate resource, material and information sharing among consortium members, which includes collaborators in academia and industry. The 2023 annual symposium was a hybrid meeting held in Washington DC on 26–27 September 2023 concurrent with the virtual attendance, with oral and poster presentations and discussions centered on microbiome-related topics within five broad thematic areas: 1) Environmental Microbiome Characterization; 2) Microbiome Analysis; 3) Human Microbiome Characterization; 4) Microbiome Engineering; and 5) In Vitro and In Vivo Microbiome Models. Collectively, the symposium provided an update on the scope of current DoD and DoD-affiliated microbiome research efforts and fostered collaborative opportunities. This report summarizes the presentations and outcomes of the 7th annual TSMC symposium.

Abstract Investigation of visual illusions helps us understand how we process visual information. For example, face pareidolia, the misperception of illusory faces in objects, could be used to understand how we process real faces. However, it remains unclear whether this illusion emerges from errors in face detection or from slower, cognitive processes. Here, our logic is straightforward; if examples of face pareidolia activate the mechanisms that rapidly detect faces in visual environments, then participants will look at objects more quickly when the objects also contain illusory faces. To test this hypothesis, we sampled continuous eye movements during a fast saccadic choice task—participants were required to select either faces or food items. During this task, pairs of stimuli were positioned close to the initial fixation point or further away, in the periphery. As expected, the participants were faster to look at face targets than food targets. Importantly, we also discovered an advantage for food items with illusory faces but, this advantage was limited to the peripheral condition. These findings are among the first to demonstrate that the face pareidolia illusion persists in the periphery and, thus, it is likely to be a consequence of erroneous face detection.

Abstract Background Previous literature on cutaneous squamous cell carcinoma (cSCC) suggests that the incidence, rate of metastasis, and tumour severity of cSCC are higher in immunosuppressed patients than in immunocompetent patients. However, current literature lacks an extensive comparison of cSCC clinical characteristics and outcomes in immunosuppressed patients. Objectives We compared cSCC tumour characteristics and disease‐related outcomes to help guide the clinical management of immunosuppressed patients. Methods We conducted a retrospective review of histopathologic and clinical data from 935 cSCC cases (19.5% immunosuppressed) from the Mayo Clinic. Results Immunosuppression was associated with younger age (69.3 vs. 74.8 years old, p

Abstract Background Disparities in cervical cancer screening rates among marginalized groups is a driver of inequalities in cervical cancer. Self‐sampling for human papillomavirus (HPV) testing is a newly emerging alternative to clinician‐performed testing to screen for cervical cancer, and has high potential to reduce screening barriers in under‐screened and marginalized groups. We study the acceptability in of HPV self‐sampling and informational materials among Black/African American, Hispanic/Spanish speaking, American Indian/Alaska Native and transgender/nonbinary populations. Methods We conducted qualitative interviews with patients, ages 30–65, who were Black/African American, Hispanic, American Indian, and/or transgender/nonbinary individuals assigned female at birth. Telephone interviews were conducted in English or Spanish. Patients did not complete the test, rather were asked about the attractiveness, comprehensibility, and acceptability of the HPV self‐test, instructions, and messaging. Results Among 23 completed interviews (5 American Indian/Alaska Native, 7 Hispanic [2 bilingual, 5 Spanish‐speaking], 5 Black/African American, and 6 transgender/nonbinary), patients from all groups thought the test was straightforward and convenient, and they would complete the test at home or in clinic. The transgender/nonbinary patients preferred at‐home testing. American Indian and transgender/nonbinary patients liked that the test might avoid pain, discomfort, and invasiveness. All patients liked the letter and instructions. All groups had specific suggestions for making the materials more culturally acceptable. Conclusions The HPV self‐test and the instructions and materials for use were acceptable for a diverse group of patients. Tailored outreach and messaging should be considered to reduce screening disparities among groups that have been historically underserved by the medical system.

Abstract Most people can conjure images and sounds that they experience in their minds. There are, however, marked individual differences. Some people report that they cannot generate imagined sensory experiences at all (aphantasics) and others report that they have unusually intense imagined experiences (hyper-phantasics). These individual differences have been linked to activity in sensory brain regions, driven by feedback. We would therefore expect imagined experiences to be associated with specific frequencies of oscillatory brain activity, as these can be a hallmark of neural interactions within and across regions of the brain. Replicating a number of other studies, relative to a Resting-State we find that the act of engaging in auditory or in visual imagery is linked to reductions in the power of oscillatory brain activity across a broad range of frequencies, with prominent peaks in the alpha band (8–12 Hz). This oscillatory activity, however, did not predict individual differences in the subjective intensity of imagined experiences. For audio imagery, these were rather predicted by reductions within the theta (6–9 Hz) and gamma (33–38 Hz) bands, and by increases in beta (15–17 Hz) band activity. For visual imagery these were predicted by reductions in lower (14–16 Hz) and upper (29–32 Hz) beta band activity, and by an increase in mid-beta band (24–26 Hz) activity. Our data suggest that there is sufficient ground truth in the subjective reports people use to describe the intensity of their imagined sensory experiences to allow these to be linked to the power of distinct rhythms of brain activity. In future, we hope to combine this approach with better measures of the subjective intensity of imagined sensory experiences to provide a clearer picture of individual differences in the subjective intensity of imagined experiences, and of why these eventuate.

ABSTRACTWe conducted a comparative evaluation of the FDA-cleared Simplexa GBS Direct and ARIES GBS molecular assays for the detection of Streptococcus agalactiae (Group B Streptococcus, GBS) in 386 prospectively collected, broth-enriched vaginal/rectal swab specimens. The sensitivity of each test was 96.2% and specificity was ≥98.7% when compared to a combined direct and enriched culture method using chromogenic culture medium. A total of four specimens were called positive by both molecular assays but negative by culture, likely representing specimens with a low burden of GBS in these specimens. Two specimens were reported positive by culture but negative by both molecular assays. One of these specimens demonstrated atypically colored colonies on chromogenic agar; the other yielded typically colored colonies only observed after broth enrichment. Our data demonstrate equivalent performance of Simplexa and ARIES molecular assays for the detection of GBS in clinical specimens.IMPORTANCEClinical laboratories often face decisions regarding which of the multiple available molecular platforms would best fit their needs based on cost, workflow, menu, and diagnostic performance. Therefore, objective clinical comparisons of similar molecular tests are valuable resources to aid these decisions. We provide a clinical comparison of two FDA-cleared tests to routine culture and to each other that can be used by clinical laboratories when determining which of the available molecular platforms would best fit their laboratory in terms of workflow, cost, and performance.

Abstract The Tri-Service Microbiome Consortium (TSMC) was founded to enhance collaboration, coordination, and communication of microbiome research among DoD organizations and to facilitate resource, material and information sharing amongst consortium members, which includes collaborators in academia and industry. The 6th Annual TSMC Symposium was a hybrid meeting held in Fairlee, Vermont on 27–28 September 2022 with presentations and discussions centered on microbiome-related topics within seven broad thematic areas: (1) Human Microbiomes: Stress Response; (2) Microbiome Analysis & Surveillance; (3) Human Microbiomes Enablers & Engineering; (4) Human Microbiomes: Countermeasures; (5) Human Microbiomes Discovery - Earth & Space; (6) Environmental Micro & Myco-biome; and (7) Environmental Microbiome Analysis & Engineering. Collectively, the symposium provided an update on the scope of current DoD microbiome research efforts, highlighted innovative research being done in academia and industry that can be leveraged by the DoD, and fostered collaborative opportunities. This report summarizes the activities and outcomes from the 6th annual TSMC symposium.

Steep Cone Geyser is a unique geothermal feature in Yellowstone National Park (YNP), Wyoming, actively gushing silicon-rich fluids along outflow channels possessing living and actively silicifying microbial biomats. To assess the geomicrobial dynamics occurring temporally and spatially at Steep Cone, samples were collected at discrete locations along one of Steep Cone’s outflow channels for both microbial community composition and aqueous geochemistry analysis during field campaigns in 2010, 2018, 2019, and 2020. Geochemical analysis characterized Steep Cone as an oligotrophic, surface boiling, silicious, alkaline-chloride thermal feature with consistent dissolved inorganic carbon and total sulfur concentrations down the outflow channel ranging from 4.59 ± 0.11 to 4.26 ± 0.07 mM and 189.7 ± 7.2 to 204.7 ± 3.55 μM, respectively. Furthermore, geochemistry remained relatively stable temporally with consistently detectable analytes displaying a relative standard deviation

Abstract One of the seminal findings of cognitive neuroscience is that the power of occipital alpha-band (~ 10 Hz) brain waves is increased when peoples’ eyes are closed, rather than open. This has encouraged the view that alpha oscillations are a default dynamic, to which the visual brain returns in the absence of input. Accordingly, we might be unable to increase the power of alpha oscillations when the eyes are closed, above the level that would normally ensue when people close their eyes. Here we report counter evidence. We used electroencephalography (EEG) to record brain activity when people had their eyes open and closed, both before and after they had adapted to radial motion. The increase in alpha power when people closed their eyes was increased by prior adaptation to a broad range of radial motion speeds. This effect was greatest for 10 Hz motion, but robust for other frequencies (and especially 7.5 Hz). This discredits a persistent entrainment of activity at the adaptation frequency as an explanation for our findings. Our data show that the power of occipital alpha-band brain waves can be increased by motion sensitive visual processes that persist when the eyes are closed. Consequently, we suggest that the power of these brain waves is, at least in part, an index of the degree to which visual brain activity is being subjected to inhibition. This is increased when people close their eyes, but can be even further increased by pre-adaptation to radial motion.

Royal jelly and honey are two substances produced successively by the worker bee caste. Modern proteomics approaches have been used to explore the protein component of each substance independently, but to date none have quantitatively compared the protein profile of honey and royal jelly directly. Sequential window acquisition of all theoretical fragment-ion spectra mass spectrometry (SWATH-MS) was used to compare protein quantities of bee origin in mānuka and clover honey to royal jelly. Two analysis techniques identified 76 proteins in total. Peptide intensity was directly compared for a subset of 31 proteins that were identified with high confidence, and the relative changes in protein abundance were compared between each honey type and royal jelly. Major Royal Jelly Proteins (MRJPs) had similar profiles in both honeys, except MRJP6, which was significantly more abundant in clover honey. Proteins involved in nectar metabolism were more abundant in honey than in royal jelly as expected. However, the trend revealed a potential catalytic role for MRJP6 in clover honey and a nectar- or honey-specific role for uncharacterised protein LOC408608. The abundance of MRJP6 in mānuka honey was equivalent to royal jelly suggesting a potential effect of nectar type on expression of this protein. Data are available via ProteomeXchange with identifier PXD038889.

There is a growing appreciation that the interaction between diet, the gut microbiota and the immune system contribute to the development and progression of inflammatory bowel disease (IBD). A mounting body of scientific evidence suggests that high-fat diets exacerbate IBD; however, there is a lack of information on how specific types of fat impact colitis. The Mediterranean diet (MD) is considered a health-promoting diet containing approximately 40% total fat. It is not known if the blend of fats found in the MD contributes to its beneficial protective effects. Mice deficient in the mucin 2 gene (Muc 2−/−) were weaned to 40% fat, isocaloric, isonitrogenous diets. We compared the MD fat blend (high monounsaturated, 2:1 n-6:n-3 polyunsaturated and moderate saturated fat) to diets composed of corn oil (CO, n-6 polyunsaturated-rich), olive oil (monounsaturated-rich) or milk fat (MF, saturated-rich) on spontaneous colitis development in Muc2−/− mice. The MD resulted in lower clinical and histopathological scores and induced tolerogenic CD103+ CD11b+ dendritic, Th22 and IL-17+ IL-22+ cells necessary for intestinal barrier repair. The MD was associated with beneficial microbes and associated with higher cecal acetic acid levels negatively correlated with colitogenic microbes like Akkermansia muciniphila. In contrast, CO showed a higher prevalence of mucin-degraders including A. muciniphila and Enterobacteriaceae, which have been associated with colitis. A dietary blend of fats mimicking the MD, reduces disease activity, inflammation-related biomarkers and improves metabolic parameters in the Muc2−/− mouse model. Our findings suggest that the MD fat blend could be incorporated into a maintenance diet for colitis.

Abstract Prediction is a core function of the human visual system. Contemporary research suggests the brain builds predictive internal models of the world to facilitate interactions with our dynamic environment. Here, we wanted to examine the behavioural and neurological consequences of disrupting a core property of peoples’ internal models, using naturalistic stimuli. We had people view videos of basketball and asked them to track the moving ball and predict jump shot outcomes, all while we recorded eye movements and brain activity. To disrupt people’s predictive internal models, we inverted footage on half the trials, so dynamics were inconsistent with how movements should be shaped by gravity. When viewing upright videos people were better at predicting shot outcomes, at tracking the ball position, and they had enhanced alpha-band oscillatory activity in occipital brain regions. The advantage for predicting upright shot outcomes scaled with improvements in ball tracking and occipital alpha-band activity. Occipital alpha-band activity has been linked to selective attention and spatially-mapped inhibitions of visual brain activity. We propose that when people have a more accurate predictive model of the environment, they can more easily parse what is relevant, allowing them to better target irrelevant positions for suppression—resulting in both better predictive performance and in neural markers of inhibited information processing.

Necrolytic acral erythema (NAE) is a relatively newly described dermatologic disease that is often associated with hepatitis C virus (HCV). Oral zinc therapy is a successful treatment; however, therapy is often delayed due to misdiagnosis. There are limited reports of NAE in the literature. This paper presents a case of NAE in a 68-year-old male with untreated HCV, whose NAE was diagnosed and treated as recurrent cellulitis for 12 years. He had low serum zinc and elevated serum glucagon levels. Elevated glucagon is not often reported in NAE, but the patient’s CT abdomen was negative, ruling out glucagonoma and necrolytic migratory erythema. He improved with oral zinc replacement and was referred to the hepatology department for HCV treatment. This paper additionally presents a review of the literature for NAE cases.

Articulography and functional neuroimaging are two major tools for studying the neurobiology of speech production. Until recently, however, it has generally not been possible to use both in the same experimental setup because of technical incompatibilities between the two methodologies. Here we describe results from a novel articulography system dubbed Magneto-articulography for the Assessment of Speech Kinematics (MASK), which we used to derive kinematic profiles of oro-facial movements during speech. MASK was used to characterize speech kinematics in two healthy adults, and the results were compared to measurements from a separate participant with a conventional Electromagnetic Articulography (EMA) system. Analyses targeted the gestural landmarks of reiterated utterances /ipa/, /api/ and /pataka/. The results demonstrate that MASK reliably characterizes key kinematic and movement coordination parameters of speech motor control. Since these parameters are intrinsically registered in time with concurrent magnetoencephalographic (MEG) measurements of neuromotor brain activity, this methodology paves the way for innovative cross-disciplinary studies of the neuromotor control of human speech production, speech development, and speech motor disorders.

Transient simulation of complex converter topologies is a challenging problem, especially in detailed analysis tools like SPICE. Transistor models presented for SPICE are often evaluated by accuracy, with less consideration for the computational cost of model elements. In order to optimize models for application simulations, this research quantifies the relative simulation performance of modeling approaches and contextualizes the results with regard to accuracy. It is well established that the primary contributor to semiconductor dynamic behavior is the voltage-dependent interelectrode capacitances. Therefore, this study isolates these model components to resolve their influence on model accuracy and run-time. Both the voltage-dependencies modeled, and the mathematic formulation chosen strongly influence the accuracy of interelectrode capacitance models. In addition to these factors, the specific implementation chosen within SPICE also determines simulation performance. Through careful evaluation of these factors, this study offers specific recommendations for optimal implementations of interelectrode capacitances in SPICE.

Most health care approaches to understanding social ills are rooted in strain or ecological models. Strain models assume that the impact of poor physical health operates through the individual, that it is the individual suffering from poor health who engages in social ills as a means of adapting, and that the impact of poor health is rather direct and immediate. Meanwhile, ecological approaches of health acknowledge how poor health may impact others and the collective, but poorly account for the case in which this is not so, leaving unexplained the many instances of people who are in poor health but remain actively engaged with their communities and preserve relationships that nurture trust, shared norms, and cooperation. To rectify this problem, we introduce the concept of “compulsive immobility”: the situation in which those in poor health are compelled to stay indoors and refrain from community socialization. We argue that compulsive immobility mediates the relationship between poor physical health and collective efficacy, suggesting that illness, specifically to a point of physical immobility (e.g., bedridden), enables poor health to detract from collective efficacy. This allows scholars to both acknowledge how poor health may impact the individual and community, while specifying the mechanism through which it operates. To support our claim, we draw on GSS data to examine the relationship among poor health, health-related immobility, and collective efficacy. Our results provide empirical support for our argument, revealing that general health conditions influenced the level of generalized trust directly and indirectly through compulsive immobility. We conclude with suggestions on how compulsive immobility might impact neighborhood crime and propose ways through which subsequent research may refine and further test compulsive immobility as a mediator between poor health and collective efficacy.

Transient simulation of complex converter topologies is a challenging problem, especially in detailed analysis tools like SPICE. Much of the recent literature on SPICE transistor modeling ignores the requirements of application designers and instead emphasizes detail, physical accuracy, and complexity. While these advancements greatly improve model accuracy, they also serve to increase computational complexity, making the resulting models less attractive to application designers. While some authors depart from this trend and present models which emphasize simulation speed, their results and analysis are limited to qualitative observation. This research develops a methodology to quantify the computational cost of model features and competitively benchmark models against each other. Additionally, it reviews recently published SiC mosfet models and presents a trade study on several candidate models likely to fare well in complex application simulations. Finally, this study also identifies key considerations which should be carried forward into future model design.

Campylobacter jejuni infection is a leading cause of foodborne disease, common to children, adult travelers, and military populations in low- to middle-income countries. In the absence of a licensed vaccine, efforts to evaluate prophylactic agents are underway. The prophylactic efficacy of a twice-daily, 550 mg dose of the antibiotic rifaximin demonstrated no efficacy against campylobacteriosis in a controlled human infection model (CHIM); however, samples from the CHIM study were utilized to assess how the human gut microbiome responds to C. jejuni infection, and if a ‘protective’ microbiota exists in study participants not developing campylobacteriosis. Statistically significant, but minor, differences in study participant beta diversity were identified during the challenge period (p = 0.002, R2 = 0.042), but no significant differences were otherwise observed. Pre-challenge alpha diversity was elevated in study participants who did not develop campylobacteriosis compared to those who did (p < 0.001), but alpha diversity declined in all study participants from the pre-challenge period to post-discharge. Our work provides insight into gut microbiome shifts observed during a C. jejuni CHIM and following antibiotic treatment. This study utilized a high dose of 1.7 x 105 colony-forming units of C. jejuni; future work could include CHIM studies performed with inocula more closely mimicking natural exposure as well as field studies involving naturally-occurring enteric infections.

Objectives: The susceptibility-vessel-sign (SVS) allows thrombus visualization, length estimation and composition, and it may impact reperfusion during mechanical thrombectomy (MT). SVS can also describe thrombus shape in the occluded artery: in the straight M1-segment (S-shaped), or in an angulated/traversing a bifurcation segment (A-shaped). We determined whether SVS clot shape influenced reperfusion and outcomes after MT for proximal middle-cerebral-artery (M1) occlusions.Methods: Between May 2015 and March 2018, consecutive patients who underwent MT at one comprehensive stroke center and who had a baseline MRI with a T2* sequence were included. Clinical, procedural and radiographic data, including clot shape on SVS [angulated/bifurcation (A-SVS) vs. straight (S-SVS)] and length were assessed. Primary outcome was successful reperfusion (TICI 2b-3). Secondary outcome were MT complication rates, MT reperfusion time, and clinical outcome at 90-days. Predictors of outcome were assessed with univariate and multivariate analyses.Results: A total of 62 patients were included. 56% (35/62) had an A-SVS. Clots were significantly longer in the A-SVS group (19 mm vs. 8 mm p = 0.0002). Groups were otherwise well-matched with regard to baseline characteristics. There was a significantly lower rate of successful reperfusion in the A-SVS cohort (83%) compared to the S-SVS cohort (96%) in multivariable analysis [OR 0.04 (95% CI, 0.002–0.58), p = 0.02]. There was no significant difference in long term clinical outcome between groups.Conclusion: Clot shape as determined on T2* imaging, in patients presenting with M1 occlusion appears to be a predictor of successful reperfusion after MT. Angulated and bifurcating clots are associated with poorer rates of successful reperfusion.

Objectives: Baseline-core-infarct volume is a critical factor in patient selection and outcome in acute ischemic stroke (AIS) before mechanical thrombectomy (MT). We determined whether oxygen extraction efficiency and arterial collaterals, two different physiologic components of the cerebral ischemic cascade, interacted to modulate baseline-core-infarct volume in patients with AIS-LVO undergoing MT triage.Methods: Between January 2015 and March 2018, consecutive patients with an AIS and M1 occlusion considered for MT with a baseline MRI and perfusion-imaging were included. Variables such as baseline-core-infarct volume [mL], arterial collaterals (HIR: TMax > 10 s volume/TMax > 6 s), high oxygen extraction (HOE, presence of the brush-sign on T2*) were assessed. A linear-regression was used to test the interaction of HOE and HIR with baseline-core-infarct volume, after including potential confounding variables.Results: We included 103 patients. Median age was 70 (58–78), and 63% were female. Median baseline-core-infarct volume was 32 ml (IQR 8–74.5). Seventy six patients (74%) had HOE. In a multivariate analysis both favorable HIR collaterals (p = 0.02) and HOE (p = 0.038) were associated with lower baseline-core-infarct volume. However, HOE significantly interacted with HIR (p = 0.01) to predict baseline-core-infarct volume, favorable collaterals (low HIR) with HOE was associated with small baseline-core-infarct whereas patients with poor collaterals (high HIR) and HOE had large baseline-core-infarct.Conclusion: While HOE under effective collateral blood-flow has the lowest baseline-core-infarct volume of all patients, the protective effect of HOE reverses under poor collateral blood-flow and may be a maladaptive response to ischemic stroke as measured by core infarctions in AIS-LVO patients undergoing MT triage.

Traveler’s diarrhea (TD) is a recurrent and significant issue for many travelers including the military. While many known enteric pathogens exist that are causative agents of diarrhea, our gut microbiome may also play a role in TD susceptibility. To this end, we conducted a pilot study of the microbiome of warfighters prior to- and after deployment overseas to identify marker taxa relevant to TD. This initial study utilized full-length 16S rRNA gene sequencing to provide additional taxonomic resolution toward identifying predictive taxa.16S rRNA analyses of pre- and post-deployment fecal samples identified multiple marker taxa as significantly differentially abundant in subjects that reported diarrhea, including Weissella, Butyrivibrio, Corynebacterium, uncultivated Erysipelotrichaceae, Jeotgallibaca, unclassified Ktedonobacteriaceae, Leptolinea, and uncultivated Ruminiococcaceae. The ability to identify TD risk prior to travel will inform prevention and mitigation strategies to influence diarrhea susceptibility while traveling.

ABSTRACT Biological sulfur cycling in polar, low-temperature ecosystems is an understudied phenomenon in part due to difficulty of access and the dynamic nature of glacial environments. One such environment where sulfur cycling is known to play an important role in microbial metabolisms is located at Borup Fiord Pass (BFP) in the Canadian High Arctic. Here, transient springs emerge from ice near the terminus of a glacier, creating a large area of proglacial aufeis (spring-derived ice) that is often covered in bright yellow/white sulfur, sulfate, and carbonate mineral precipitates accompanied by a strong odor of hydrogen sulfide. Metagenomic sequencing of samples from multiple sites and of various sample types across the BFP glacial system produced 31 metagenome-assembled genomes (MAGs) that were queried for sulfur, nitrogen, and carbon cycling/metabolism genes. An abundance of sulfur cycling genes was widespread across the isolated MAGs and sample metagenomes taxonomically associated with the bacterial classes Alphaproteobacteria and Gammaproteobacteria and Campylobacteria (formerly the Epsilonproteobacteria). This corroborates previous research from BFP implicating Campylobacteria as the primary class responsible for sulfur oxidation; however, data reported here suggested putative sulfur oxidation by organisms in both the alphaproteobacterial and gammaproteobacterial classes that was not predicted by previous work. These findings indicate that in low-temperature, sulfur-based environments, functional redundancy may be a key mechanism that microorganisms use to enable coexistence whenever energy is limited and/or focused by redox chemistry. IMPORTANCE A unique environment at Borup Fiord Pass is characterized by a sulfur-enriched glacial ecosystem in the low-temperature Canadian High Arctic. BFP represents one of the best terrestrial analog sites for studying icy, sulfur-rich worlds outside our own, such as Europa and Mars. The site also allows investigation of sulfur-based microbial metabolisms in cold environments here on Earth. Here, we report whole-genome sequencing data that suggest that sulfur cycling metabolisms at BFP are more widely used across bacterial taxa than predicted. From our analyses, the metabolic capability of sulfur oxidation among multiple community members appears likely due to functional redundancy present in their genomes. Functional redundancy, with respect to sulfur-oxidation at the BFP sulfur-ice environment, may indicate that this dynamic ecosystem hosts microorganisms that are able to use multiple sulfur electron donors alongside other metabolic pathways, including those for carbon and nitrogen.

Recent evidence shows that hippocampal theta oscillations, usually linked to memory and navigation, are also observed during online language processing, suggesting a shared neurophysiological mechanism between language and memory. However, it remains to be established what specific roles hippocampal theta oscillations may play in language, and whether and how theta mediates the communication between the hippocampus and the perisylvian cortical areas, generally thought to support language processing. With whole-head magnetoencephalographic (MEG) recordings, the present study investigated these questions with two experiments. Using a violation paradigm, extensively used for studying neural underpinnings of different aspects of linguistic processing, we found increased theta power (4–8 ​Hz) in the hippocampal formation, when participants read a semantically incorrect vs. correct sentence ending. Such a pattern of results was replicated using different sentence stimuli in another cohort of participants. Importantly, no significant hippocampal theta power increase was found when participants read a semantically correct but syntactically incorrect sentence ending vs. a correct sentence ending. These findings may suggest that hippocampal theta oscillations are specifically linked to lexical-semantic related processing, and not general information processing in sentence reading. Furthermore, we found significantly transient theta phase coupling between the hippocampus and the left superior temporal gyrus, a hub area of the cortical network for language comprehension. This transient theta phase coupling may provide an important channel that links the memory and language systems for the generation of sentence meaning. Overall, these findings help specify the role of hippocampal theta in language, and provide a novel neurophysiological mechanism at the network level that may support the interface between memory and language.

Impairment of microglial functions, such as phagocytosis and/or dysregulation of immune responses, has been implicated as an underlying factor involved in the pathogenesis of various neurodegenerative disorders. Our previous studies have demonstrated that long intergenic noncoding RNA (lincRNA)-Cox2 expression is influenced by nuclear factor κB (NF-κB) signaling and serves as a coactivator of transcriptional factors to regulate the expression of a vast array of immune-related genes in microglia. Extracellular vesicles (EVs) have been recognized as primary facilitators of cell-to-cell communication and cellular regulation. Herein, we show that EVs derived from astrocytes exposed to morphine can be taken up by microglial endosomes, leading, in turn, to activation of Toll-like receptor 7 (TLR7) with a subsequent upregulation of lincRNA-Cox2 expression, ultimately resulting in impaired microglial phagocytosis. This was further validated in vivo, wherein inhibition of microglial phagocytic activity was also observed in brain slices isolated from morphine-administrated mice compared with control mice. Additionally, we also showed that intranasal delivery of EVs containing lincRNA-Cox2 siRNA (small interfering RNA) was able to restore microglial phagocytic activity in mice administered morphine. These findings have ramifications for the development of EV-loaded RNA-based therapeutics for the treatment of various disorders involving functional impairment of microglia. Keywords: phagocytosis, drug abuse, morphine, extracellular vesicle, exosome, lincRNA, microglia, CNS

Electrophysiological studies of adults indicate that brain activity is enhanced during viewing of repeated faces, at a latency of about 250 ms after the onset of the face (M250/N250). The present study aimed to determine if this effect was also present in preschool-aged children, whose brain activity was measured in a custom-sized pediatric MEG system. The results showed that, unlike adults, face repetition did not show any significant modulation of M250 amplitude in children; however children’s M250 latencies were significantly faster for repeated than non-repeated faces. Dynamic causal modelling (DCM) of the M250 in both age groups tested the effects of face repetition within the core face network including the occipital face area (OFA), the fusiform face area (FFA), and the superior temporal sulcus (STS). DCM revealed that repetition of identical faces altered both forward and backward connections in children and adults; however the modulations involved inputs to both FFA and OFA in adults but only to OFA in children. These findings suggest that the amplitude-insensitivity of the immature M250 may be due to a weaker connection between the FFA and lower visual areas. Keywords: MEG, Face recognition, Repetition, DCM, M250, M170

Evidence is accumulating that similar cognitive resources are engaged to process syntactic structure in music and language. Congenital amusia – a neurodevelopmental disorder that primarily affects music perception, including musical syntax – provides a special opportunity to understand the nature of this overlap. Using electroencephalography (EEG), we investigated whether individuals with congenital amusia have parallel deficits in processing language syntax in comparison to control participants. Twelve amusic participants (eight females) and 12 control participants (eight females) were presented melodies in one session, and spoken sentences in another session, both of which had syntactic-congruent and -incongruent stimuli. They were asked to complete a music-related and a language-related task that were irrelevant to the syntactic incongruities. Our results show that amusic participants exhibit impairments in the early stages of both music- and language-syntactic processing. Specifically, we found that two event-related potential (ERP) components – namely Early Right Anterior Negativity (ERAN) and Left Anterior Negativity (LAN), associated with music- and language-syntactic processing respectively, were absent in the amusia group. However, at later processing stages, amusics showed similar brain responses as controls to syntactic incongruities in both music and language. This was reflected in a normal N5 in response to melodies and a normal P600 to spoken sentences. Notably, amusics' parallel music- and language-syntactic impairments were not accompanied by deficits in semantic processing (indexed by normal N400 in response to semantic incongruities). Together, our findings provide further evidence for shared music and language syntactic processing, particularly at early stages of processing. Keywords: Music, Language, Syntax, Congenital amusia, ERP

Renewable fuels hold great promise for the future yet their susceptibility to biodegradation and subsequent corrosion represents a challenge that needs to be directly assessed. Biodiesel is a renewable fuel that is widely used as a substitute or extender for petroleum diesel and is composed of a mixture of fatty acid methyl esters derived from plant or animal fats. Biodiesel can be blended up to 20% v/v with ultra-low sulfur diesel (i.e., B20) and used interchangeably with diesel engines and infrastructure. The addition of biodiesel, however, has been linked to increased susceptibility to biodegradation. Microorganisms proliferating via degradation of biodiesel blends have been linked to microbiologically influenced corrosion in the laboratory, but not measured directly in storage tanks (i.e., in situ). To measure in situ microbial proliferation, fuel degradation and microbially influenced corrosion, we conducted a yearlong study of B20 storage tanks in operation at two locations, identified the microorganisms associated with fuel fouling, and measured in situ corrosion. The bacterial populations were more diverse than the fungal populations, and largely unique to each location. The bacterial populations included members of the Acetobacteraceae, Clostridiaceae, and Proteobacteria. The abundant Eukaryotes at both locations consisted of the same taxa, including a filamentous fungus within the family Trichocomaceae, not yet widely recognized as a contaminant of petroleum fuels, and the Saccharomycetaceae family of yeasts. Increases in the absolute and relative abundances of the Trichocomaceae were correlated with significant, visible fouling and pitting corrosion. This study identified the relationship between fouling of B20 with increased rates of corrosion and the microorganisms responsible, largely at the bottom of the sampled storage tanks. To our knowledge this is the first in situ study of this scale incorporating community and corrosion measurements in an active biodiesel storage environment.

Naked mole-rats (Heterocephalus glaber) are subterranean rodents that utilize their incisors for feeding, chisel-tooth digging of complex tunnel systems, social interactions, and defense in their eusocial colony structure. Previous studies have shown that naked mole-rats have morphological and anatomical adaptations that predict strong bite forces, namely, skulls that are relatively tall and wide, in addition to impressive masticatory musculature. However, no studies to date have directly measured bite force in this species or analyzed the relationship between bite force and social caste. In the current study, we assessed adult naked mole-rat maximum bite force in relation to body mass, in addition to considering each animal’s position within the eusocial hierarchy (i.e., dominant versus subordinate). Each animal was permitted to freely interact with a piezo-resistive bite force sensor. Our results showed that bite force was correlated with body mass in subordinate but not in dominant naked mole-rats, and that subordinate animals exhibited a shorter latency in producing their first bite. Maximum bite force was significantly influenced by caste. In comparing bite force with available data from previous studies across 82 additional mammalian species, subordinate naked mole-rats exhibited a bite force that was 65% higher than predicted for their body size, comparable to Tasmanian devils and exceeding bite force values for all of the carnivorans included for comparison. These results supported the hypothesis that the naked mole-rat’s bite force would exceed predictions based on body size due to the behavioral importance and specialization of the naked mole-rat incisors. This study provides insight into the differences in bite force across species, and the significant role that social and ecological factors might play in the evolutionary relationship between bite force performance and underlying anatomical structures.

ObjectivesRapid and accurate mold identification is critical for guiding therapy for mold infections. MALDI-TOF MS has been widely adopted for bacterial and yeast identification; however, few clinical laboratories have applied this technology for routine mold identification due to limited database availability and lack of standardized processes. Here, we evaluated the versatility of the NIH Mold Database in a multicenter evaluation.MethodsThe NIH Mold Database was evaluated by eight US academic centers using a solid media extraction method and a challenge set of 80 clinical mold isolates. Multiple instrument parameters important for spectra optimization were evaluated, leading to the development of two specialized acquisition programs (NIH method and the Alternate-B method).ResultsA wide range in performance (33–77%) was initially observed across the eight centers when routine spectral acquisition parameters were applied. Use of the NIH or the Alternate-B specialized acquisition programs, which are different than those used routinely for bacterial and yeast spectral acquisition (MBT_AutoX), in combination with optimized instrument maintenance, improved performance, illustrating that acquisition parameters may be one of the key limiting variable in achieving successful performance.ConclusionSuccessful mold identification using the NIH Database for MALDI-TOF MS on Biotyper systems was demonstrated across multiple institutions for the first time following identification of critical program parameters combined with instrument optimization. This significantly advances our potential to implement MALDI-TOF MS for mold identification across many institutions. Because instrument variability is inevitable, development of an instrument performance standard specific for mold spectral acquisition is suggested to improve reproducibility across instruments.

To achieve maximal growth, cells must manage a massive economy of ribosomal proteins (r-proteins) and RNAs (rRNAs) to produce thousands of ribosomes every minute. Although ribosomes are essential in all cells, natural disruptions to ribosome biogenesis lead to heterogeneous phenotypes. Here, we model these perturbations in Saccharomyces cerevisiae and show that challenges to ribosome biogenesis result in acute loss of proteostasis. Imbalances in the synthesis of r-proteins and rRNAs lead to the rapid aggregation of newly synthesized orphan r-proteins and compromise essential cellular processes, which cells alleviate by activating proteostasis genes. Exogenously bolstering the proteostasis network increases cellular fitness in the face of challenges to ribosome assembly, demonstrating the direct contribution of orphan r-proteins to cellular phenotypes. We propose that ribosome assembly is a key vulnerability of proteostasis maintenance in proliferating cells that may be compromised by diverse genetic, environmental, and xenobiotic perturbations that generate orphan r-proteins.

Drawing from Emmanuel Levinas’s philosophical ethics and Paul Celan’s dialogical poetics, this article interrogates the impossible memorial and ethical demands that literary responses to the Holocaust place upon their readers. While Levinas reveals our position as summoned to radical responsibility, Celan shows us how that responsibility plays out in the form of ethical reading. By attending to the imperative commands found in Celan’s longest poem, “Engführung”, this article demonstrates how Holocaust literature memorializes the Shoah through an invocation of Levinasian ethics and the concept of the immemorial—that which exceeds memory. Following the discussion of Levinas, Celan, and “Engführung”, I turn to Primo Levi’s “Shema”, a paradigmatic text that likewise directly challenges us, calling us into question as readers during the moment of reading and demanding an attentiveness to the text that proves beyond our ability to deliver. Throughout, I aim to show how dialogical memory enables us to better comprehend the ethical burden we encounter in the literary texts of the Holocaust.

An isolate of the soil fungus Scopulariopsis brevicaulis was identified from the surface of female winter ticks (Dermacentor albipictus) collected from recently dead moose (Alces alces) calves in New Hampshire in the northeastern United States. It was the sole isolate, and it matched with 98% nucITS similarity (molecular systematics Blast match) to S. brevicaulis species from soil and other tick species. Inoculation of tick larvae and eggs with 108 spores/mL + 0.05% Tween (aqueous inoculum) resulted in mortality, reduced survival time, and recovery of S. brevicaulis from within tick tissues. Rapid water loss and death from dehydration were the pathogenic consequences of the fungal infection. Three entomopathogenic fungal isolates from laboratory culture (Beauveria bassiana, B. caledonica, and Metarhizium anisopliae) inoculated concurrently at the same dose, were slightly less pathogenic to eggs than larvae of winter ticks. We conclude that S. brevicaulis imposes a limitation on the freeliving stages of the winter tick population in specific environmental conditions, but commercial fungal treatments as used in local situations to control ticks, are impractical as a means of controlling winter tick density across moose habitats.

Conventional water resources are not sufficient in many regions to meet the needs of growing populations. Due to cyclical weather cycles, drought, and climate change, water stress has increased worldwide including in Southern California, which serves as a model for regions that integrate reuse of wastewater for both potable and non-potable use. The Orange County Water District (OCWD) Advanced Water Purification Facility (AWPF) is a highly engineered system designed to treat and produce up to 100 million gallons per day (MGD) of purified water from a municipal wastewater source for potable reuse. Routine facility microbial water quality analysis is limited to standard indicators at this and similar facilities. Given recent advances in high throughput DNA sequencing techniques, complete microbial profiling of communities in water samples is now possible. By using 16S/18S rRNA gene sequencing, metagenomic and metatranscriptomic sequencing coupled to a highly accurate identification method along with 16S rRNA gene qPCR, we describe a detailed view of the total microbial community throughout the facility. The total bacterial load of the water at stages of the treatment train ranged from 3.02 × 106 copies in source, unchlorinated wastewater feed to 5.49 × 101 copies of 16S rRNA gene/mL after treatment (consisting of microfiltration, reverse osmosis, and ultraviolet/advanced oxidation). Microbial diversity and load decreased by several orders of magnitude after microfiltration and reverse osmosis treatment, falling to almost non-detectable levels that more closely resembled controls of molecular grade laboratory water than the biomass detected in the source water. The presence of antibiotic resistance genes and viruses was also greatly reduced. Overall, system design performance was achieved, and comprehensive microbial community analysis was found to enable a more complete characterization of the water/wastewater microbial signature.

Microbial mats are found in a variety of modern environments, with evidence for their presence as old as the Archean. There is much debate about the rates and conditions of processes that eventually lithify and preserve mats as microbialites. Here, we apply novel tracer experiments to quantify both mat biomass addition and the formation of CaCO3. Microbial mats from Little Hot Creek (LHC), California, contain calcium carbonate that formed within multiple mat layers, and thus constitute a good test case to investigate the relationship between the rate of microbial mat growth and carbonate precipitation. The laminated LHC mats were divided into four layers via color and fabric, and waters within and above the mat were collected to determine their carbonate saturation states. Samples of the microbial mat were also collected for 16S rRNA analysis of microbial communities in each layer. Rates of carbonate precipitation and carbon fixation were measured in the laboratory by incubating homogenized samples from each mat layer with δ13C-labeled HCO3- for 24 h. Comparing these rates with those from experimental controls, poisoned with NaN3 and HgCl2, allowed for differences in biogenic and abiogenic precipitation to be determined. Carbon fixation rates were highest in the top layer of the mat (0.17% new organic carbon/day), which also contained the most phototrophs. Isotope-labeled carbonate was precipitated in all four layers of living and poisoned mat samples. In the top layer, the precipitation rate in living mat samples was negligible although abiotic precipitation occurred. In contrast, the bottom three layers exhibited biologically enhanced carbonate precipitation. The lack of correlation between rates of carbon fixation and biogenic carbonate precipitation suggests that processes other than autotrophy may play more significant roles in the preservation of mats as microbialites.

Hot spring environments can create physical and chemical gradients favorable for unique microbial life. They can also include authigenic mineral precipitates that may preserve signs of biological activity on Earth and possibly other planets. The abiogenic or biogenic origins of such precipitates can be difficult to discern, therefore a better understanding of mineral formation processes is critical for the accurate interpretation of biosignatures from hot springs. Little Hot Creek (LHC) is a hot spring complex located in the Long Valley Caldera, California, that contains mineral precipitates composed of a carbonate base (largely submerged) topped by amorphous silica (largely emergent). The precipitates occur in close association with microbial mats and biofilms. Geological, geochemical, and microbiological data are consistent with mineral formation via degassing and evaporation rather than direct microbial involvement. However, the microfabric of the silica portion is stromatolitic in nature (i.e., wavy and finely laminated), suggesting that abiogenic mineralization has the potential to preserve textural biosignatures. Although geochemical and petrographic evidence suggests the calcite base was precipitated via abiogenic processes, endolithic microbial communities modified the structure of the calcite crystals, producing a textural biosignature. Our results reveal that even when mineral precipitation is largely abiogenic, the potential to preserve biosignatures in hot spring settings is high. The features found in the LHC structures may provide insight into the biogenicity of ancient Earth and extraterrestrial rocks.

Hippocampal rhythms are believed to support crucial cognitive processes including memory, navigation, and language. Due to the location of the hippocampus deep in the brain, studying hippocampal rhythms using non-invasive magnetoencephalography (MEG) recordings has generally been assumed to be methodologically challenging. However, with the advent of whole-head MEG systems in the 1990s and development of advanced source localization techniques, simulation and empirical studies have provided evidence that human hippocampal signals can be sensed by MEG and reliably reconstructed by source localization algorithms. This paper systematically reviews simulation studies and empirical evidence of the current capacities and limitations of MEG “deep source imaging” of the human hippocampus. Overall, these studies confirm that MEG provides a unique avenue to investigate human hippocampal rhythms in cognition, and can bridge the gap between animal studies and human hippocampal research, as well as elucidate the functional role and the behavioral correlates of human hippocampal oscillations.

In recent years, librarians in various sectors have been moving forward a conversation on the distinct information needs and information-seeking behavior of our lesbian, gay, bisexual, trans, and queer (LGBTQ) patrons and how well the profession recognizes and meets those needs. Health sciences librarianship has been slower than other areas of the profession in creating an evidence base covering the needs of its LGBTQ patrons, with, until recently, only very limited literature on this subject. LGBTQ health sciences librarianship is now starting to attract new interest, with librarians working together to bring this emerging specialization to the attention of the broader professional community. In this paper, the authors report on a dedicated panel discussion that took place at the 2016 joint annual meeting of the Medical Library Association and Canadian Health Libraries Association/Association des bibliothèques de la santé du Canada in Toronto, Ontario, Canada; discuss subsequent reflections; and highlight the emerging role for health sciences librarians in providing culturally competent services to the LGBTQ population. Recommendations are also provided for establishing a tool kit for LGBTQ health sciences librarianship from which librarians can draw. We conclude by highlighting the importance of critically reflective practice in health sciences librarianship in the context of LGBTQ health information.

Pharmacological interventions aimed at improving outcomes in type 2 diabetes and achieving normoglycaemia, including insulin therapy, are increasingly common, despite the potential for substantial side effects. Carbohydrate-restricted diets that result in increased ketogenesis have effectively been used to improve insulin resistance, a fundamental feature of type 2 diabetes. In addition, limited evidence suggests that states of ketogenesis may also improve β-cell function in type 2 diabetics. Considering how little is known regarding the effects of ketones on β-cell function, we sought to determine the specific effects of β-Hydroxybutyrate (βHB) on pancreatic β-cell physiology and mitochondrial function. βHB treatment increased β-cell survival and proliferation, while also increasing mitochondrial mass, respiration and adenosine triphosphate (ATP) production. Despite these improvements, were unable to detect an increase in β-cell or islet insulin production and secretion. Collectively, these findings have two implications. Firstly, they indicate that β-cells have improved survival and proliferation in the midst of βHB, the circulating form of ketones. Secondly, insulin secretion does not appear to be directly related to apparent improvements in mitochondrial function and cellular proliferation.

Smartphones and tablets are finding their way into healthcare delivery to the extent that mobile health (mHealth) has become an identifiable field within eHealth. In prior work, a mobile app to document chronic wounds and wound care, specifically pressure ulcers (bedsores) was developed for Android smartphones and tablets. One feature of the mobile app allowed users to take images of the wound using the smartphone or tablet's integrated camera. In a user trial with nurses at a personal care home, this feature emerged as a key benefit of the mobile app. This paper developed image analysis algorithms that facilitate noncontact measurements of irregularly shaped images (e.g., wounds), where the image is taken with a sole smartphone or tablet camera. The image analysis relies on the sensors integrated in the smartphone or tablet with no auxiliary or add-on instrumentation on the device. Three approaches to image analysis were developed and evaluated: 1) computing depth using autofocus data; 2) a custom sensor fusion of inertial sensors and feature tracking in a video stream; and 3) a custom pinch/zoom approach. The pinch/zoom approach demonstrated the strongest potential and thus developed into a fully functional prototype complete with a measurement mechanism. While image analysis is a very well developed field, this paper contributes to image analysis applications and implementation in mHealth, specifically for wound care.

Objective. Oral inflammatory pathologies are linked to increased oxidative stress, thereby partly explaining their relevance in the etiology of systemic disorders. The purpose of this work was to determine the degree to which LPS from Porphyromonas gingivalis, the primary pathogen related to oral inflammation, altered gingival mitochondrial function and reactive oxygen species generation. Methods. Human gingival fibroblast (HGF-1) cells were treated with lipopolysaccharide of P. gingivalis. Mitochondrial function was determined via high-resolution respirometry. Results. LPS-treated HGF-1 cells had significantly higher mitochondrial complex IV and higher rates of mitochondrial respiration. However, this failed to translate into greater ATP production, as ATP production was paradoxically diminished with LPS treatment. Nevertheless, production of the reactive H2O2 was elevated with LPS treatment. Conclusions. LPS elicits an increase in gingival cell mitochondria content, with a subsequent increase in reactive oxygen species production (i.e., H2O2), despite a paradoxical reduction in ATP generation. These findings provide an insight into the nature of oxidative stress in oral inflammatory pathologies.

Modern high-speed atomic force microscopes generate significant quantities of data in a short amount of time. Each image in the sequence has to be processed quickly and accurately in order to obtain a true representation of the sample and its changes over time. This paper presents an automated, adaptive algorithm for the required processing of AFM images. The algorithm adaptively corrects for both common one-dimensional distortions as well as the most common two-dimensional distortions. This method uses an iterative thresholded processing algorithm for rapid and accurate separation of background and surface topography. This separation prevents artificial bias from topographic features and ensures the best possible coherence between the different images in a sequence. This method is equally applicable to all channels of AFM data, and can process images in seconds.

Musical imagery is a relatively unexplored area, partly because of deficiencies in existing experimental paradigms, which are often difficult, unreliable, or do not provide objective measures of performance. Here we describe a novel protocol, the Pitch Imagery Arrow Task (PIAT), which induces and trains pitch imagery in both musicians and non-musicians. Given a tonal context and an initial pitch sequence, arrows are displayed to elicit a scale-step sequence of imagined pitches, and participants indicate whether the final imagined tone matches an audible probe. It is a staircase design that accommodates individual differences in musical experience and imagery ability. This new protocol was used to investigate the roles that musical expertise, self-reported auditory vividness and mental control play in imagery performance. Performance on the task was significantly better for participants who employed a musical imagery strategy compared to participants who used an alternative cognitive strategy and positively correlated with scores on the Control subscale from the Bucknell Auditory Imagery Scale (BAIS). Multiple regression analysis revealed that Imagery performance accuracy was best predicted by a combination of strategy use and scores on the Vividness subscale of BAIS. These results confirm that competent performance on the PIAT requires active musical imagery and is very difficult to achieve using alternative cognitive strategies. Auditory vividness and mental control were more important than musical experience in the ability to perform manipulation of pitch imagery.

The prompt and accurate identification of bacterial pathogens is fundamental to patient health and outcome. Recent advances in matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) have revolutionized bacterial identification in the clinical laboratory, but uniform incorporation of this technology in the U.S. market has been delayed by a lack of FDA-cleared systems. In this study, we conducted a multicenter evaluation of the MALDI Biotyper CA (MBT-CA) System (Bruker Daltonics Inc, Billerica, MA) for the identification of aerobic gram-negative bacteria as part of a 510(k) submission to the FDA. A total of 2,263 aerobic gram negative bacterial isolates were tested representing 23 genera and 61 species. Isolates were collected from various clinical sources and results obtained from the MBT-CA System were compared to DNA sequencing and/or biochemical testing. Isolates that failed to report as a "high confidence species ID" [log(score) ≥2.00] were re-tested using an extraction method. The MBT-CA System identified 96.8% and 3.1% of isolates with either a "high confidence" or a "low confidence" [log(score) value between 1.70 and