Low Oxygen Concentration Reduces Neisseria gonorrhoeae Susceptibility to Resazurin.

Neisseria gonorrhoeae has developed resistance to every antibiotic currently approved for the treatment of gonorrhea, prompting the development of new therapies. The phenoxazine dye resazurin exhibits robust antimicrobial activity against N. gonorrhoeae in vitro but fails to limit vaginal colonization by N. gonorrhoeae in a mouse model. The lack of in vivo efficacy may be due to oxygen limitation as in vitro susceptibility assays with resazurin are conducted under atmospheric oxygen while a microaerophilic environment is present in the vagina. Here, we utilized broth microdilution assays to determine the susceptibility of N. gonorrhoeae to resazurin under low and atmospheric oxygen conditions. The minimal inhibitory concentration of resazurin for multiple N. gonorrhoeae clinical isolates was significantly higher under low oxygen. This effect was specific to resazurin as N. gonorrhoeae was equally susceptible to other antibiotics under low and atmospheric oxygen conditions. The reduced susceptibility of N. gonorrhoeae to resazurin under low oxygen was largely attributed to reduced oxidative stress, as the addition of antioxidants under atmospheric oxygen mimicked the reduced susceptibility to resazurin observed under low oxygen. Together, these data suggest oxygen concentration is an important factor to consider when evaluating the efficacy of new antibiotics against N. gonorrhoeae in vitro.

Infection Rates After Aseptic Revision Total Hip Arthroplasty With Extended Oral Antibiotic Prophylaxis.

INTRODUCTION: Few US studies have investigated the efficacy of extended oral antibiotic prophylaxis (EOAP) in the prevention of periprosthetic joint infection (PJI) after aseptic revision total hip arthroplasty (R-THA). This study compared PJI rates in aseptic R-THA performed with EOAP with PJI rates in published studies of aseptic R-THA patients not receiving EOAP. METHODS: Prospectively documented data on 127 consecutive aseptic R-THAs were retrospectively reviewed. Evidence-based perioperative infection prevention protocols were used, and all patients were discharged on 7-day EOAP. Superficial and deep infections at 30 and 90 days postoperatively and at mean latest follow-up of 27.8 months were statistically compared with all US studies reporting the prevalence of PJI after aseptic R-THA. Complications related to EOAP within 120 days of the index procedure also are reported. RESULTS: No superficial or deep infections were observed at 30 and 90 days postoperatively when 7-day postdischarge EOAP was used. Superficial and deep infection rates were 1.57% (two patients) and 3.15% (four patients) at mean latest follow-up, respectively. Comparisons with published 30-day PJI rates of 1.37% ( P = 0.423) and 1.85% ( P = 0.257) were not statistically significant. Two of four comparisons with published 90-day PJI rates of 3.43% ( P = 0.027) and 5.74% ( P = 0.001) were statistically different. The deep PJI rate of 3.15% at mean latest follow-up was significantly lower than two of three published rates at equivalent follow-up including 10.10% ( P = 0.009) and 9.12% ( P = 0.041). No antibiotic-related complications were observed within 120 days of the index procedure. DISCUSSION: Study findings possibly support the use of EOAP after aseptic R-THA to prevent catastrophic PJI with revision implants, indicating that the efficacy of EOAP cannot be definitively ruled-in or ruled-out based on available evidence.

Cysteine dependence of Lactobacillus iners is a potential therapeutic target for vaginal microbiota modulation.

Vaginal microbiota composition affects many facets of reproductive health. Lactobacillus iners-dominated microbial communities are associated with poorer outcomes, including higher risk of bacterial vaginosis (BV), compared with vaginal microbiota rich in L. crispatus. Unfortunately, standard-of-care metronidazole therapy for BV typically results in dominance of L. iners, probably contributing to post-treatment relapse. Here we generate an L. iners isolate collection comprising 34 previously unreported isolates from 14 South African women with and without BV and 4 previously unreported isolates from 3 US women. We also report an associated genome catalogue comprising 1,218 vaginal Lactobacillus isolate genomes and metagenome-assembled genomes from >300 women across 4 continents. We show that, unlike L. crispatus, L. iners growth is dependent on L-cysteine in vitro and we trace this phenotype to the absence of canonical cysteine biosynthesis pathways and a restricted repertoire of cysteine-related transport mechanisms. We further show that cysteine concentrations in cervicovaginal lavage samples correlate with Lactobacillus abundance in vivo and that cystine uptake inhibitors selectively inhibit L. iners growth in vitro. Combining an inhibitor with metronidazole promotes L. crispatus dominance of defined BV-like communities in vitro by suppressing L. iners growth. Our findings enable a better understanding of L. iners biology and suggest candidate treatments to modulate the vaginal microbiota to improve reproductive health for women globally.

Role of dipA and pilD in Francisella tularensis Susceptibility to Resazurin.

The phenoxazine dye resazurin exhibits bactericidal activity against the Gram-negative pathogens Francisella tularensis and Neisseria gonorrhoeae. One resazurin derivative, resorufin pentyl ether, significantly reduces vaginal colonization by Neisseria gonorrhoeae in a mouse model of infection. The narrow spectrum of bacteria susceptible to resazurin and its derivatives suggests these compounds have a novel mode of action. To identify potential targets of resazurin and mechanisms of resistance, we isolated mutants of F. tularensis subsp. holarctica live vaccine strain (LVS) exhibiting reduced susceptibility to resazurin and performed whole genome sequencing. The genes pilD (FTL_0959) and dipA (FTL_1306) were mutated in half of the 46 resazurin-resistant (RZR) strains sequenced. Complementation of select RZR LVS isolates with wild-type dipA or pilD partially restored sensitivity to resazurin. To further characterize the role of dipA and pilD in resazurin susceptibility, a dipA deletion mutant, ΔdipA, and pilD disruption mutant, FTL_0959d, were generated. Both mutants were less sensitive to killing by resazurin compared to wild-type LVS with phenotypes similar to the spontaneous resazurin-resistant mutants. This study identified a novel role for two genes dipA and pilD in F. tularensis susceptibility to resazurin.

Biomechanical Comparison of a First- and a Second-Generation All-Soft Suture Glenoid Anchor.

BACKGROUND: All-soft tissue suture anchors provide advantages of decreased removal of bone and decreased glenoid volume occupied compared with traditional tap or screw-in suture anchors. Previous published data have led to biomechanical concerns with the use of first-generation all-soft suture anchors. PURPOSE/HYPOTHESIS: The purpose of this study was to evaluate the load to 2-mm displacement and ultimate load to failure of a second-generation all-soft suture anchor, compared with a first-generation anchor and a traditional PEEK (polyether ether ketone) anchor. The null hypothesis was that the newer second-generation anchor will demonstrate no difference in loads to 2-mm displacement after cycling compared with first-generation all-soft suture anchors. STUDY DESIGN: Controlled laboratory study. METHODS: Twenty human cadaveric glenoids were utilized to create 97 total suture anchor sites, and 1 of 3 anchors were randomized and placed into each site: (1) first-generation all-soft suture anchor (Juggerknot; Biomet), (2) second-generation all-soft suture anchor (Suturefix; Smith & Nephew), and (3) a control PEEK anchor (Bioraptor; Smith & Nephew). After initial cyclic loading, load to 2 mm of displacement and ultimate load to failure were measured for each anchor. RESULTS: After cyclic loading, the load to 2-mm displacement was significantly less in first-generation anchors compared with controls (P < .01). However, the load to 2-mm displacement was significantly greater in second-generation anchors compared with controls (P < .01). There was no difference in ultimate load to failure between the first- and second-generation all-soft suture anchors (P > .05). CONCLUSION: The newer generation all-soft suture anchors with a theoretically more rigid construct and deployment configuration demonstrate biomechanical characteristics (specifically, with load to 2-mm displacement after cyclic loading) that are improved over first-generation all-soft suture anchors and similar to a traditional solid tap-in anchor. The configuration of these newer generation all-soft suture anchors appears to mitigate the biomechanical concerns of decreased load to failure with first-generation all-soft tissue suture anchors. CLINICAL RELEVANCE: The theoretical advantages of all-soft anchors may be particularly valuable in revision surgery or in cases where multiple anchors are being placed into a small anatomic area.

Subject position affects EEG magnitudes.

EEG (electroencephalography) has been used for decades in thousands of research studies and is today a routine clinical tool despite the small magnitude of measured scalp potentials. It is widely accepted that the currents originating in the brain are strongly influenced by the high resistivity of skull bone, but it is less well known that the thin layer of CSF (cerebrospinal fluid) has perhaps an even more important effect on EEG scalp magnitude by spatially blurring the signals. Here it is shown that brain shift and the resulting small changes in CSF layer thickness, induced by changing the subject's position, have a significant effect on EEG signal magnitudes in several standard visual paradigms. For spatially incoherent high-frequency activity the effect produced by switching from prone to supine can be dramatic, increasing occipital signal power by several times for some subjects (on average 80%). MRI measurements showed that the occipital CSF layer between the brain and skull decreases by approximately 30% in thickness when a subject moves from prone to supine position. A multiple dipole model demonstrated that this can indeed lead to occipital EEG signal power increases in the same direction and order of magnitude as those observed here. These results suggest that future EEG studies should control for subjects' posture, and that some studies may consider placing their subjects into the most favorable position for the experiment. These findings also imply that special consideration should be given to EEG measurements from subjects with brain atrophy due to normal aging or neurodegenerative diseases, since the resulting increase in CSF layer thickness could profoundly decrease scalp potential measurements.

Methods for extra-low voltage transcranial direct current stimulation: current and time dependent impedance decreases.

OBJECTIVE: Though tDCS is well tolerated, it is desirable to further limit the voltage applied for additional safety factors and optimized device design. We investigated the minimum voltage required for tDCS using 1.5 and 2.5mA. METHODS: Impedance data has been collected prior to, during and after 18 tDCS sessions, using 1.5mA and 2.5mA tDCS currents and three different test current magnitudes. Data was pooled and tested for differences using t-tests, corrected for multiple comparisons. Average impedance data was fitted into a RLC circuit model with additional double integrator. RESULTS: We report that the impedance drop during tDCS initiation significantly reduces the voltage compliance required to achieve the target current (14.5V for 1.5mA, 18.5V for 2.5mA). Data was well approximated by a 4th order linear impedance model. CONCLUSION: In addition to indicating the feasibility of reduced voltage tDCS, we propose an extra-low voltage "Limited Total Energy" approach where stimulation is continued at voltage compliance allowing time for impedance to decrease and target current to be reached. SIGNIFICANCE: Reduced-voltage and Limited Total Energy tDCS are viable approaches towards more protective and robust tDCS protocols.

A pilot study on effects of 4×1 high-definition tDCS on motor cortex excitability.

High-Definition transcranial Direct Current Stimulation (HD-tDCS) using specialized small electrodes has been proposed as a focal, non-invasive neuromodulatory technique. Here we provide the first evidence of a change in cortical excitability after HD-tDCS of the motor cortex, using TMS motor evoked potential (MEP) as the measure of excitability. Stimulation for 20 minutes at 1 mA with an anode centered over the hand area of the motor cortex and four surrounding return electrodes (anodal 4×1 montage) produced a significant increase in MEP amplitude and variability after stimulation, compared to sham stimulation. Stimulation was well tolerated by all subjects with adverse effects limited to transient sensation under the electrodes. A high-resolution computational model confirmed predictions of increased focality using the 4×1 HD tDCS montage compared to conventional tDCS. Simulations also indicated that variability in placement of the center electrode relative to the location of the target (central sulcus) could account for increasing variability. These results provide support for the careful use of this technique where focal tDCS is desired.

Fast reconstruction and prediction of frozen flow turbulence based on structured Kalman filtering.

Efficient and optimal prediction of frozen flow turbulence using the complete observation history of the wavefront sensor is an important issue in adaptive optics for large ground-based telescopes. At least for the sake of error budgeting and algorithm performance, the evaluation of an accurate estimate of the optimal performance of a particular adaptive optics configuration is important. However, due to the large number of grid points, high sampling rates, and the non-rationality of the turbulence power spectral density, the computational complexity of the optimal predictor is huge. This paper shows how a structure in the frozen flow propagation can be exploited to obtain a state-space innovation model with a particular sparsity structure. This sparsity structure enables one to efficiently compute a structured Kalman filter. By simulation it is shown that the performance can be improved and the computational complexity can be reduced in comparison with auto-regressive predictors of low order.

An inquiry-based approach to teaching research design: asking the right questions.

To describe an inquiry-based learning activity and discuss the advantages of using such activities to demonstrate research design to undergraduate psychology students the authors presented students with the research problem of eyewitness memory recall and asked students to solve the problem, using the information provided by the course. Working in small groups, students reviewed eyewitness studies and designed their own research materials, including a videotaped theft and a CD-ROM version of the eyewitness survey and potential suspect line-up. Poststudy interviews showed that the students enjoyed the activity and felt that they had a better understanding of experimental research design than they had before the exercise. It is proposed that this activity may be a useful hands-on tool for instructors who have difficulty teaching students fundamental topics such as literature reviews, hypothesis testing, reliability and validity of research measures, and experimental design.