Blunting specific T-dependent antibody responses with engineered "decoy" B cells.

Antibody inhibitors pose an ongoing challenge to the treatment of subjects with inherited protein deficiency disorders, limiting the efficacy of both protein replacement therapy and corrective gene therapy. Beyond their central role as producers of serum antibody, B cells also exhibit many unique properties that could be exploited in cell therapy applications, notably including antigen-specific recognition and the linked capacity for antigen presentation. Here we employed CRISPR-Cas9 to demonstrate that ex vivo antigen-primed Blimp1-knockout "decoy" B cells, incapable of differentiation into plasma cells, participated in and downregulated host antigen-specific humoral responses after adoptive transfer. Following ex vivo antigen pulse, adoptively transferred high-affinity antigen-specific decoy B cells were diverted into germinal centers en masse, thereby reducing participation by endogenous antigen-specific B cells in T-dependent humoral responses and suppressing both cognate and linked antigen-specific immunoglobulin (Ig)G following immunization with conjugated antigen. This effect was dose-dependent and, importantly, did not impact concurrent unrelated antibody responses. We demonstrated the therapeutic potential of this approach by treating factor VIII (FVIII)-knockout mice with antigen-pulsed decoy B cells prior to immunization with an FVIII conjugate protein, thereby blunting the production of serum FVIII-specific IgG by an order of magnitude as well as reducing the proportion of animals exhibiting functional FVIII inhibition by 6-fold.

Bridging the Gap in Graduate Medical Education: A Longitudinal Analysis of Medical and Fellowship Director Experiences in Telehealth.

Background With the evolving nature of telehealth services being supported across graduate medical programs, understanding changing attitudes among program directors is essential for training future physicians. Objective This 5-year longitudinal survey analysis provides details regarding the knowledge, skills, competency, and confidence levels present among program directors and the ways telehealth training (didactic and experiential) supports resident education. Methods A longitudinal observation survey was sent to 77 program directors (48% completion) from an academic health system. Data were compared from survey responses from 2016 to 2021 with additional information collected about current training processes in 2021. Paired sample t-tests, quantitative data analysis, and qualitative thematic analysis results are reported based on a convenience sampling of all program and fellowship directors. Results Results show that while telehealth knowledge and use increased among program director faculty, systemic concerns about the feasibility of telehealth from a time and cost perspective remain high. In 2016 and 2021, most of those surveyed (28.9% and 37.1%, respectively) were concerned about reimbursement. Directors remain committed to telehealth resident education, with over 60% stating that it is "essential to future practice". The qualitative thematic analysis highlighted the need for additional resources and support to conduct telehealth and the variability within disciplines of the use, therefore modeling, of telehealth in clinical settings. Conclusions The overall utilization of telehealth by graduate medical education programs has increased along with the continued need for training to prepare residents for current and future practice.

Immunization with a tri-antigen syphilis vaccine significantly attenuates chancre development, reduces bacterial load, and inhibits dissemination of Treponema pallidum.

Syphilis continues to be a significant public health concern worldwide. The disease is endemic in many low- and middle-income countries, and rates have risen sharply in high-income countries over the last decade. The continued prevalence of infectious and congenital syphilis worldwide highlights the need for the development of an effective syphilis vaccine to complement public health measures for syphilis control. The complex, multi-stage course of syphilis infection necessitates a holistic approach to the development of an effective vaccine, in which immunization prevents both the localized stage of infection (typified by the highly infectious chancre) and the disseminated stages of infection (typified by the secondary rash, neurosyphilis, and destructive tertiary lesions, as well as congenital syphilis). Inhibiting development of the infectious chancre would reduce transmission thus providing community- level protection, while preventing dissemination would provide individual-level protection by reducing serious sequelae and may also provide community level protection by reducing shedding during secondary syphilis. In the current study we build upon prior investigations which demonstrated that immunizations with individual, well characterized T. pallidum TprK, TprC, and Tp0751 peptides elicits partial protection against infection in the animal model. Specifically, we show here that immunization with a TprC/TprK/Tp0751 tri-antigen cocktail protects animals from progressive syphilis lesions and substantially inhibits dissemination of the infection.

Telesimulation to Improve Critical Decision-Making in Prehospital Airway Management: A Feasibility Study.

OBJECTIVES: Telesimulation, in which learners and evaluators use technology to connect remotely to simulation-based learning activities, is effective for skills and decision-making review. Historical models in which learners are colocated with the simulation equipment have inherent issues, especially for emergency medical services (EMS) providers. This feasibility study placed the evaluators in the simulation center, whereas the learners were at a distance steering the scenario evolution through telehealth technologies. METHODS: Volunteer EMS providers across South Carolina with varying levels of training and experience completed difficult airway management scenarios focused on clinical decision making. The program consisted of pre- and postexperience examinations, a lecture, and increasingly complicated simulations using high-fidelity mannequins that were facilitated by local trainers under the direction of remote trainees. Audio and video content, including vital signs and cardiac monitoring, were live streamed. Participants worked in two-person teams with lead providers on each scenario clinically assessing and managing cases of anaphylaxis. Data were collected from the simulations using Laerdal software, as well as examination and survey results. RESULTS: A total of 24 participants completed all of the elements of the training. Trends toward improvement in times to bag-mask ventilation and initial epinephrine administration were noted. Average cognitive test scores increased by 9.6%, and learners reported improved comfort with simulation (75%, P ≥ 0.0001) and videoconferencing (83%, P ≥ 0.0001). They also reported high degrees of comfort with intubation (73.3%) following the training. CONCLUSIONS: This method of telesimulation appears to be a viable addition to continuing EMS education and may address access issues for some providers.

A Spray-Dried Combination of Capreomycin and CPZEN-45 for Inhaled Tuberculosis Therapy.

Tuberculosis (TB) remains the single most serious infectious disease attributable to a single-causative organism. A variety of drugs have been evaluated for pulmonary delivery as dry powders: capreomycin sulfate has shown efficacy and was safely delivered by inhalation at high doses to human volunteers, whereas CPZEN-45 is a new drug that has also been shown to kill resistant TB. The studies here combine these drugs-acting by different mechanisms-as components of single particles by spray-drying, yielding a new combination drug therapy. The spray-dried combination powder was prepared in an aerodynamic particle size range suitable for pulmonary delivery. Physicochemical storage stability was demonstrated for a period of 6 months. The spray-dried combination powders of capreomycin and CPZEN-45 have only moderate affinity for mucin, indicating that delivered drug will not be bound by these mucins in the lung and available for microbicidal effects. The pharmacokinetics of disposition in guinea pigs demonstrated high local concentrations of drug following direct administration to the lungs and subsequent systemic bioavailability. Further studies are required to demonstrate the in vivo efficacy of the combination to confirm the therapeutic potential of this novel combination.

Regulation of Bacterial Gene Expression by Protease-Alleviated Spatial Sequestration (PASS).

In natural microbial systems, conditional spatial sequestration of transcription factors enables cells to respond rapidly to changes in their environment or intracellular state by releasing presynthesized regulatory proteins. Although such a mechanism may be useful for engineering synthetic biology technologies ranging from cell-based biosensors to biosynthetic platforms, to date it remains unknown how or whether such conditional spatial sequestration may be engineered. In particular, based upon seemingly contradictory reports in the literature, it is not clear whether subcellular spatial localization of a transcription factor within the cytoplasm is sufficient to preclude regulation of cognate promoters on plasmid-borne or chromosomal loci. Here, we describe a modular, orthogonal platform for investigating and implementing this mechanism using protease-alleviated spatial sequestration (PASS). In this system, expression of an exogenous protease mediates the proteolytic release of engineered transcriptional regulators from the inner face of the Escherichia coli cytoplasmic membrane. We demonstrate that PASS mediates robust, conditional regulation of either transcriptional repression, via tetR, or transcriptional activation, by the λ phage CI protein. This work provides new insights into a biologically important facet of microbial gene expression and establishes a new strategy for engineering conditional transcriptional regulation for the microbial synthetic biology toolbox.

Antibacterial activity of some folklore medicinal plants from South India.

Antibacterial activity and phytochemical tests of eight whole plant methanol extracts belonging to family Euphorbiaceae were evaluated. In agar well diffusion assay the diameter of inhibition zones ranged from 3-13 mm. Phyllanthus emblica showed maximum activity of 13 mm. The MIC and MBC observed were 30-140 mcg/ml and 40-160 mcg/ml, respectively. P. piscatorum and P. emblica showed the lowest MIC (30 mcg/ml), P. emblica the lowest MBC (40 mcg/ml) and thus an effective inhibitor of the tested bacteria. Alkaloids, saponins and tannins were detected in 7 out of 8 tested plants.

Improving attitudes regarding the elderly population: the effects of information and reinforcement for change.

PURPOSE: Altering negative attitudes associated with ageism may be possible by giving people accurate information about older people in conjunction with reinforcement for change. DESIGN AND METHODS: Ninety-nine college students (35 men, 63 women; mean age = 20 years, SD = 2.78) participated in one of three groups: information only, information plus an innocuous discussion group, and information plus a reinforcement-to-change discussion group. The participants' attitudes toward elderly people were measured before, immediately after the intervention, and at a one-month follow-up. Changes in attitudes across groups and time were analyzed using analysis of variance (ANOVA) and t tests. RESULTS: Information alone produced initial improvements in attitudes in all groups; however, only the group members who received additional reinforcement for change maintained positive attitude changes at one-month follow-up. IMPLICATIONS: This study supports the premise that negative attitudes toward older people are amendable; however, the new attitude may be lost without reinforcement for change.

Treatment of the cutaneous pain of acute herpes zoster with 9% lidocaine (base) in petrolatum/paraffin ointment.

BACKGROUND: Treatment of the pain of acute herpes zoster by local anesthetic injections has drawbacks. Topical percutaneous local anesthesia (TPLA) may offer another strategy of providing regional analgesia in affected patients. OBJECTIVE: We evaluate the analgesic efficacy and safety of 9% (wt/vol) lidocaine (base) in petrolatum/paraffin ointment in patients with acute herpes zoster. METHODS: Ointment was applied to the affected skin of 22 patients. Pain, tenderness, sensitivity to pinprick and cold, and blood lidocaine concentration were measured repeatedly during a 20-hour interval and intermittently thereafter. RESULTS: Mean pain, tenderness, and cutaneous sensation scores were reduced at measurements taken from 4 to 20 hours after ointment application (p < 0.05), but not every patient obtained relief. No patient had local skin irritation or systemic toxic effects related to the local anesthetic. CONCLUSIONS: TPLA is a promising therapy for control of cutaneous pain of acute herpes zoster. Controlled studies should be performed to prove efficacy, determine optimal TPLA formulation, and define dosage limits.