Mpox infection protects against re-challenge in rhesus macaques.

The mpox outbreak of 2022-2023 involved rapid global spread in men who have sex with men. We infected 18 rhesus macaques with mpox by the intravenous, intradermal, and intrarectal routes and observed robust antibody and T cell responses following all three routes of infection. Numerous skin lesions and high plasma viral loads were observed following intravenous and intradermal infection. Skin lesions peaked on day 10 and resolved by day 28 following infection. On day 28, we re-challenged all convalescent and 3 naive animals with mpox. All convalescent animals were protected against re-challenge. Transcriptomic studies showed upregulation of innate and inflammatory responses and downregulation of collagen formation and extracellular matrix organization following challenge, as well as rapid activation of T cell and plasma cell responses following re-challenge. These data suggest key mechanistic insights into mpox pathogenesis and immunity. This macaque model should prove useful for evaluating mpox vaccines and therapeutics.

Vaccine protection against the SARS-CoV-2 Omicron variant in macaques.

The rapid spread of the SARS-CoV-2 Omicron (B.1.1.529) variant, including in highly vaccinated populations, has raised important questions about the efficacy of current vaccines. In this study, we show that the mRNA-based BNT162b2 vaccine and the adenovirus-vector-based Ad26.COV2.S vaccine provide robust protection against high-dose challenge with the SARS-CoV-2 Omicron variant in cynomolgus macaques. We vaccinated 30 macaques with homologous and heterologous prime-boost regimens with BNT162b2 and Ad26.COV2.S. Following Omicron challenge, vaccinated macaques demonstrated rapid control of virus in bronchoalveolar lavage, and most vaccinated animals also controlled virus in nasal swabs. However, 4 vaccinated animals that had moderate Omicron-neutralizing antibody titers and undetectable Omicron CD8+ T cell responses failed to control virus in the upper respiratory tract. Moreover, virologic control correlated with both antibody and T cell responses. These data suggest that both humoral and cellular immune responses contribute to vaccine protection against a highly mutated SARS-CoV-2 variant.

Mucosal boosting enhances vaccine protection against SARS-CoV-2 in macaques.

A limitation of current SARS-CoV-2 vaccines is that they provide minimal protection against infection with current Omicron subvariants1,2, although they still provide protection against severe disease. Enhanced mucosal immunity may be required to block infection and onward transmission. Intranasal administration of current vaccines has proven inconsistent3-7, suggesting that alternative immunization strategies may be required. Here we show that intratracheal boosting with a bivalent Ad26-based SARS-CoV-2 vaccine results in substantial induction of mucosal humoral and cellular immunity and near-complete protection against SARS-CoV-2 BQ.1.1 challenge. A total of 40 previously immunized rhesus macaques were boosted with a bivalent Ad26 vaccine by the intramuscular, intranasal and intratracheal routes, or with a bivalent mRNA vaccine by the intranasal route. Ad26 boosting by the intratracheal route led to a substantial expansion of mucosal neutralizing antibodies, IgG and IgA binding antibodies, and CD8+ and CD4+ T cell responses, which exceeded those induced by Ad26 boosting by the intramuscular and intranasal routes. Intratracheal Ad26 boosting also led to robust upregulation of cytokine, natural killer, and T and B cell pathways in the lungs. After challenge with a high dose of SARS-CoV-2 BQ.1.1, intratracheal Ad26 boosting provided near-complete protection, whereas the other boosting strategies proved less effective. Protective efficacy correlated best with mucosal humoral and cellular immune responses. These data demonstrate that these immunization strategies induce robust mucosal immunity, suggesting the feasibility of developing vaccines that block respiratory viral infections.

Vaccines elicit highly conserved cellular immunity to SARS-CoV-2 Omicron.

The highly mutated SARS-CoV-2 Omicron (B.1.1.529) variant has been shown to evade a substantial fraction of neutralizing antibody responses elicited by current vaccines that encode the WA1/2020 spike protein1. Cellular immune responses, particularly CD8+ T cell responses, probably contribute to protection against severe SARS-CoV-2 infection2-6. Here we show that cellular immunity induced by current vaccines against SARS-CoV-2 is highly conserved to the SARS-CoV-2 Omicron spike protein. Individuals who received the Ad26.COV2.S or BNT162b2 vaccines demonstrated durable spike-specific CD8+ and CD4+ T cell responses, which showed extensive cross-reactivity against both the Delta and the Omicron variants, including in central and effector memory cellular subpopulations. Median Omicron spike-specific CD8+ T cell responses were 82-84% of the WA1/2020 spike-specific CD8+ T cell responses. These data provide immunological context for the observation that current vaccines still show robust protection against severe disease with the SARS-CoV-2 Omicron variant despite the substantially reduced neutralizing antibody responses7,8.

Impaired upper respiratory tract barrier function during postnatal development predisposes to invasive pneumococcal disease.

Infants are highly susceptible to invasive respiratory and gastrointestinal infections. To elucidate the age-dependent mechanism(s) that drive bacterial spread from the mucosa, we developed an infant mouse model using the prevalent pediatric respiratory pathogen, Streptococcus pneumoniae (Spn). Despite similar upper respiratory tract (URT) colonization levels, the survival rate of Spn-infected infant mice was significantly decreased compared to adults and corresponded with Spn dissemination to the bloodstream. An increased rate of pneumococcal bacteremia in early life beyond the newborn period was attributed to increased bacterial translocation across the URT barrier. Bacterial dissemination in infant mice was independent of URT monocyte or neutrophil infiltration, phagocyte-derived ROS or RNS, inflammation mediated by toll-like receptor 2 or interleukin 1 receptor signaling, or the pore-forming toxin pneumolysin. Using molecular barcoding of Spn, we found that only a minority of bacterial clones in the nasopharynx disseminated to the blood in infant mice, indicating the absence of robust URT barrier breakdown. Rather, transcriptional profiling of the URT epithelium revealed a failure of infant mice to upregulate genes involved in the tight junction pathway. Expression of many such genes was also decreased in early life in humans. Infant mice also showed increased URT barrier permeability and delayed mucociliary clearance during the first two weeks of life, which corresponded with tighter attachment of bacteria to the respiratory epithelium. Together, these results demonstrate a window of vulnerability during postnatal development when altered mucosal barrier function facilitates bacterial dissemination.

Effects of Capsular Polysaccharide amount on Pneumococcal-Host interactions.

Among the many oral streptococci, Streptococcus pneumoniae (Spn) stands out for the capacity of encapsulated strains to cause invasive infection. Spread beyond upper airways, however, is a biological dead end for the organism, raising the question of the benefits of expending energy to coat its surface in a thick layer of capsular polysaccharide (CPS). In this study, we compare mutants of two serotypes expressing different amounts of CPS and test these in murine models of colonization, invasion infection and transmission. Our analysis of the effect of CPS amount shows that Spn expresses a capsule of sufficient thickness to shield its surface from the deposition of complement and binding of antibody to underlying epitopes. While effective shielding is permissive for invasive infection, its primary contribution to the organism appears to be in the dynamics of colonization. A thicker capsule increases bacterial retention in the nasopharynx, the first event in colonization, and also impedes IL-17-dependent clearance during late colonization. Enhanced colonization is associated with increased opportunity for host-to-host transmission. Additionally, we document substantial differences in CPS amount among clinical isolates of three common serotypes. Together, our findings show that CPS amount is highly variable among Spn and could be an independent determinant affecting host interactions.

Enzymatic Characterization of a Novel HSL Family IV Esterase EstD04 from Pseudomonas sp. D01 in Mealworm Gut Microbiota.

Pseudomonas sp. D01, capable of growing in tributyrin medium, was isolated from the gut microbiota of yellow mealworm. By using in silico analyses, we discovered a hypothesized esterase encoding gene in the D01 bacterium, and its encoded protein, EstD04, was classified as a bacterial hormone-sensitive lipase (bHSL) of the type IV lipase family. The study revealed that the recombinant EstD04-His(6x) protein exhibited esterase activity and broad substrate specificity, as it was capable of hydrolyzing p-nitrophenyl derivatives with different acyl chain lengths. By using the most favorable substrate p-nitrophenyl butyrate (C4), we defined the optimal temperature and pH value for EstD04 esterase activity as 40 °C and pH 8, respectively, with a catalytic efficiency (kcat/Km) of 6.17 × 103 mM-1 s-1 at 40 °C. EstD04 demonstrated high stability between pH 8 and 10, and thus, it might be capably used as an alkaline esterase in industrial applications. The addition of Mg2+ and NH4+, as well as DMSO, could stimulate EstD04 enzyme activity. Based on bioinformatic motif analyses and tertiary structural simulation, we determined EstD04 to be a typical bHSL protein with highly conserved motifs, including a triad catalytic center (Ser160, Glu253, and His283), two cap regions, hinge sites, and an oxyanion hole, which are important for the type IV enzyme activity. Moreover, the sequence analysis suggested that the two unique discrete cap regions of EstD04 may contribute to its alkali mesophilic nature, allowing EstD04 to exhibit extremely distinct physiological properties from its evolutionarily closest esterase.

Access to naloxone at community pharmacies under the Massachusetts statewide standing order.

OBJECTIVE: This study aimed to evaluate access to, and barriers to accessing, naloxone at community pharmacies throughout Massachusetts following implementation of new legislation that requires all community pharmacies to maintain a sufficient supply for dispensing under a statewide standing order. DESIGN: From September 2018 through January 2019, we conducted a cross-sectional telephone-based survey of Massachusetts pharmacies by having an interviewer pose as a customer seeking naloxone. SETTING AND PARTICIPANTS: Community pharmacies were identified from a list of all actively licensed pharmacies provided by the Massachusetts Department of Public Health and one-half were randomly selected for inclusion. Pharmacies that were permanently closed, duplicated on the list, or closed to the general public were excluded from analysis. OUTCOME MEASURES: Rates of stocked naloxone, perceived need for identification or prescription, and pricing. RESULTS: Of the 524 pharmacies surveyed, 97.7% (n = 512) reported routinely stocking naloxone. Of those, 90.4% (n = 463) had naloxone in stock on the day of contact. Most pharmacies with naloxone in stock did not require a prescription (96.1%; n = 445); at these pharmacies, personal identification was required by 38.9% (n = 180). The average out-of-pocket naloxone nasal spray price was $128.34 ± $40.75. CONCLUSION: Nearly all Massachusetts community pharmacies routinely stock naloxone as required by state law; however, barriers remain regarding perceived need for identification and high out-of-pocket costs.

Plasma biomarkers of risk for death in a multicenter phase 3 trial with uniform transplant characteristics post-allogeneic HCT.

A phase 3 clinical trial (BMT CTN 0402) comparing tacrolimus/sirolimus (Tac/Sir) vs tacrolimus/methotrexate (Tac/Mtx) as graft-versus-host disease (GVHD) prophylaxis after matched-related allogeneic hematopoietic cell transplantation (HCT) recently showed no difference between study arms in acute GVHD-free survival. Within this setting of a prospective, multicenter study with uniform GVHD prophylaxis, conditioning regimen, and donor source, we explored the correlation of 10 previously identified biomarkers with clinical outcomes after allogeneic HCT. We measured biomarkers from plasma samples collected in 211 patients using enzyme-linked immunosorbent assay (Tac/Sir = 104, Tac/Mtx = 107). High suppression of tumorigenicity-2 (ST2) and T-cell immunoglobulin mucin-3 (TIM3) at day 28 correlated with 2-year nonrelapse mortality in multivariate analysis (P = .0050, P = .0075, respectively) and in a proportional hazards model with time-dependent covariates (adjusted hazard ratio: 2.43 [1.49-3.95], P = .0038 and 4.87 [2.53-9.34], P < .0001, respectively). High ST2 and TIM3 correlated with overall survival. Chemokine (C-X-C motif) ligand 9 (CXCL9) levels above the median were associated with chronic GVHD compared with levels below the median in a time-dependent proportional hazard analysis (P = .0069). Low L-Ficolin was associated with hepatic veno-occlusive disease (P = .0053, AUC = 0.80). We confirmed the correlation of plasma-derived proteins, previously assessed in single-center cohorts, with clinical outcomes after allogeneic HCT within this prospective, multicenter study.

Are You on the Right Platform? A Conjoint Analysis of Social Media Preferences in Aesthetic Surgery Patients.

BACKGROUND: Social media has become an indispensable tool for patients to learn about aesthetic surgery. Currently, procedure-specific patient preferences for social media platforms and content are unknown. OBJECTIVES: The authors sought to evaluate social media preferences of patients seeking aesthetic surgery. METHODS: We utilized a choice-based conjoint analysis survey to analyze the preferences of patients seeking 3 common aesthetic procedures: breast augmentation (BA), facial rejuvenation (FR), and combined breast/abdominal surgery (BAB). Participants were asked to choose among social media platforms (Facebook, Twitter, Instagram, Snapchat, Pinterest, Tumblr, YouTube), information extent (basic, moderate, comprehensive), delivery mechanism (prerecorded video, live video, photographs, text description), messenger (surgeon, nurse/clinic staff, patient), and option for interactivity (yes/no). The survey was administered using an Internet crowdsourcing service (Amazon Mechanical Turk). RESULTS: A total of 647 participants were recruited: 201 in BA, 255 in FR, and 191 in BAB. Among attributes surveyed, participants in all 3 groups (BA, FR, BAB) valued social media platform as the most important (30.9%, 33.1%, 31.4%), followed by information extent (23.1%, 22.9%, 21.6%), delivery mechanism (18.9%, 17.4%, 18%), messenger (16%, 17%, 17.2%), and interactivity (11.1%, 9.8%, 11.8%). Within these attributes, Facebook ranked as the preferred platform, with comprehensive information extent, live video as the delivery mechanism, and surgeon as the messenger as most preferred. CONCLUSIONS: The choice of social media platform is the most important factor for patients, and they indicated a preference for comprehensive information delivered by the surgeon via live video on Facebook. Our study elucidates social media usage in common aesthetic populations, which can help improve aesthetic patient outreach.

When helping helps: exploring health benefits of cancer survivors participating in for-cause physical activity events.

BACKGROUND: Over 15.5 million Americans live with cancer and 5-year survival rates have risen to 69%. Evidence supports important health benefits of regular physical activity for cancer survivors, including increased strength and quality of life, and reduced fatigue, recurrence, and mortality. However, physical activity participation among cancer survivors remains low. Cancer organizations provide various resources and support for cancer survivors, including emotional, instrumental, informational, and appraisal support. Many cancer organizations, like the LIVESTRONG Foundation, support the cancer community by sponsoring and hosting for-cause physical activity events, providing opportunities for anyone (including cancer survivors) to "help"/support those living with cancer. The concept of helping others has been positively related with wellbeing, physical activity, and multiple health behaviors for those helping. However, the role of helping others has not been examined in the context of being physically active to help others or its relationship with overall physical activity and quality of life among those helping. Therefore, we developed a path model to examine relationships between cancer survivors' (1) desire to help others with cancer, (2) physically active LIVESTRONG participation to help others, (3) regular physical activity engagement, and (4) quality of life. METHODS: In 2010, 3257 cancer survivors responded to an online survey sent to all people involved with the LIVESTRONG organization at any level. The hypothesized path model was tested using path analysis (Mplus 8). RESULTS: After list-wise deletion of missing responses, our final sample size was 3122 (61.8% female, mean age: 48.2 years [SD = 12.7]). Results indicated that the model yielded perfect fit indexes. Controlling for age, sex, income, and survivorship length, desire to help was positively related with physically active LIVESTRONG participation (ß = .11, p < .001), which was positively related with regular physical activity (ß = .30, p < .001), and regular physical activity was positively related with quality of life (ß = .194, p < .001). CONCLUSIONS: Results suggest that cancer survivors can benefit from participating in for-cause physical activity events, including more regular physical activity. Researchers need to further investigate the role of helping others when examining health behaviors and outcomes, and cancer organizations should continue encouraging cancer survivors to help others by participating in physical activity events.

A Phase l Study of a Tumor-targeted Systemic Nanodelivery System, SGT-94, in Genitourinary Cancers.

Gene therapy development has been limited by our inability to target multifocal cancer with systemic delivery. We developed a systemically administered, tumor-targeted liposomal nanodelivery complex (SGT-94) carrying a plasmid encoding RB94, a truncated form of the RB gene. In preclinical studies, RB94 showed marked cytotoxicity against tumor but not normal cells. SGT-94 was administered intravenously in a first-in-man study in metastatic genitourinary cancer. Minimal side effects were observed; dose-limiting toxicity (DLT) has not been reached in 11 evaluable patients. There was evidence of clinical activity at the 2.4 mg dose with one complete remission (CR) and one partial remission (PR). The patient in CR was retreated upon progression and had a second PR. Furthermore, there was tumor-specific targeting of the SGT-94 complex. One patient had wedge resections of two lung metastases which demonstrated RB94 expression at the DNA level by polymerase chain reaction (PCR) and at the protein level by Western blotting, with no RB94 present in normal contiguous lung. In conclusion, systemically delivered SGT-94 showed evidence of selective tumor targeting and was well tolerated with evidence of clinical activity. Additional studies are warranted to explore the activity of this drug as a single agent and in combination therapy.

What Do Our Patients Truly Want? Conjoint Analysis of an Aesthetic Plastic Surgery Practice Using Internet Crowdsourcing.

BACKGROUND: What do patients want when looking for an aesthetic surgeon? When faced with attributes like reputation, years in practice, testimonials, photos, and pricing, which is more valuable? Moreover, are attributes procedure-specific? Currently, inadequate evidence exists on which attributes are most important to patients, and to our knowledge, none on procedure-specific preferences. OBJECTIVES: First, to determine the most important attributes to breast augmentation, combined breast/abdominal surgery, and facelift patients using conjoint analysis. Second, to test the conjoint using an internet crowdsourcing service (Amazon Mechanical Turk [MTurk]). METHODS: Anonymous university members were asked, via mass electronic survey, to pick a surgeon for facelift surgery based on five attributes. Attribute importance and preference was calculated. Once pre-tested, the facelift, breast augmentation and combined breast/abdominal surgery surveys were administered worldwide to MTurk. RESULTS: The university facelift cohort valued testimonials (33.9%) as the most important, followed by photos (31.6%), reputation (18.2%), pricing (14.4%), and practice years (1.9%). MTurk breast augmentation participants valued photos (35.3%), then testimonials (33.9%), reputation (15.7%), pricing (12.2%), and practice years (3%). MTurk combined breast/abdominal surgery and facelift participants valued testimonials (38.3% and 38.1%, respectively), then photos (27.9%, 29.4%), reputation (17.5%, 15.8%), pricing (13.9%, 13.9%), practice years (2.4%, 2.8%). CONCLUSIONS: Breast augmentation patients placed higher importance on photos; combined breast/abdominal surgery and facelift patients valued testimonials. Conjoint analysis has had limited application in plastic surgery. To our knowledge, internet crowdsourcing is a novel participant recruitment method in plastic surgery. Its unique benefits include broad, diverse and anonymous participant pools, low-cost, rapid data collection, and high completion rate.

Implementation and Analysis of a Lean Six Sigma Program in Microsurgery to Improve Operative Throughput in Perforator Flap Breast Reconstruction.

PURPOSE: Perforator flaps have become a preferred method of breast reconstruction but can consume considerable resources. We examined the impact of a Six Sigma program on microsurgical breast reconstruction at an academic medical center. METHODS: Using methods developed by Motorola and General Electric, we applied critical pathway planning, workflow analysis, lean manufacturing, continuous quality improvement, and defect reduction to microsurgical breast reconstruction. Primary goals were to decrease preoperative-to-cut time and total operative time, through reduced variability and improved efficiency. Secondary goals were to reduce length of stay, complications, and reoperation. The project was divided into 3 phases: (1) Pre-Six Sigma (24 months), (2) Six Sigma (10 months), (3) and Post-Six Sigma (24 months). These periods (baseline, intervention, control) were compared by Student t test and χ analysis. RESULTS: Over a 5-year period, 112 patients underwent 168 perforator flaps for breast reconstructions, by experienced microsurgeons. Total operative time decreased from 714 to 607 minutes (P < 0.01), across the study period, with the greatest drop occurring in unilateral cases, from 672 to 498 minutes (P < 0.01). Length of stay decreased from 6.3 to 5.2 days (P = 0.01). Overall complication rates (35.9% vs 30%, not significant) and take-back rates (20.5% vs 23.9%, not significant) remained similar over the 5-year period. Physician revenue/minute increased from US $6.28 to US $7.59, whereas hospital revenue/minute increased from US $21.84 to US $25.11. CONCLUSIONS: A Six Sigma program in microsurgical breast reconstruction was associated with better operational and financial outcomes. These incremental gains were maintained over the course of the study, suggesting that these benefits were due, in part, to process improvements. However, continued reductions in total operative time and length of stay, well after the intervention period, support the possibility that "learning curve" phenomenon may have contributed to the improvement in these outcomes.

Preoperative Saline Implant Deflation in Revisional Aesthetic Breast Surgery.

BACKGROUND: Preoperative saline deflation is a clinically useful intervention in revisional breast surgery. It allows suspensory ligament recovery, reveals true glandular volume, and simplifies mastopexy markings. Presently unknown are the volumetric changes that occur after deflation. OBJECTIVES: The authors report the three-dimensional (3D) changes that occur with preoperative deflation prior to revisional breast surgery. METHODS: We reviewed available charts of revisional breast surgery patients who underwent preliminary saline implant deflation. Our protocol is deflation 4 weeks prior to revision. Three weeks following deflation, the patient is evaluated to finalize the operative plan, including the need for implants, mastopexy, and adjunctive procedures. A subset underwent 3D imaging to quantify the volumetric changes over the 3-week deflation period. RESULTS: Between 2002 and 2014, 55 patients underwent saline implant deflation prior to 57 revisional surgeries. Seventeen were revised without implants and 40 with implants. The 3D subset of 10 patients showed a mean 15.2% volume increase and 0.18 cm notch-to-nipple distance decrease over the 3 weeks following deflation and prior to definitive surgical correction. CONCLUSIONS: Breast volume increases and the notch-to-nipple distance decreases during the 3-week interval prior to reoperation. This "elastic breast recoil" occurs after the mass effect of the implant is removed, resulting in recovery of stretched suspensory ligaments and gland reexpansion. We believe 4 weeks is optimal for gland normalization. Ideal candidates include patients requiring secondary mastopexy without implants, implant downsizing in the same pocket, and secondary augmentation mastopexy. Preoperative saline deflation and 3D analyses are useful for preoperative planning in reoperative breast surgery.

Dissecting a complex chemical stress: chemogenomic profiling of plant hydrolysates.

The efficient production of biofuels from cellulosic feedstocks will require the efficient fermentation of the sugars in hydrolyzed plant material. Unfortunately, plant hydrolysates also contain many compounds that inhibit microbial growth and fermentation. We used DNA-barcoded mutant libraries to identify genes that are important for hydrolysate tolerance in both Zymomonas mobilis (44 genes) and Saccharomyces cerevisiae (99 genes). Overexpression of a Z. mobilis tolerance gene of unknown function (ZMO1875) improved its specific ethanol productivity 2.4-fold in the presence of miscanthus hydrolysate. However, a mixture of 37 hydrolysate-derived inhibitors was not sufficient to explain the fitness profile of plant hydrolysate. To deconstruct the fitness profile of hydrolysate, we profiled the 37 inhibitors against a library of Z. mobilis mutants and we modeled fitness in hydrolysate as a mixture of fitness in its components. By examining outliers in this model, we identified methylglyoxal as a previously unknown component of hydrolysate. Our work provides a general strategy to dissect how microbes respond to a complex chemical stress and should enable further engineering of hydrolysate tolerance.

Epm2a suppresses tumor growth in an immunocompromised host by inhibiting Wnt signaling.

The genetic mechanisms responsible for increased incidence of lymphoma in immunocompromised individuals have not been fully elucidated. We show that, in a line of TCR transgenic TG-B mice, an insertional mutation in one allele of the Epm2a locus and epigenetic silencing of another led to a high rate of lymphoma with early onset. Overexpressing Epm2a suppressed the growth of established tumor cells and the development of lymphoma in the TG-B mice, while specific silencing of the locus increased tumorigenesis in the immune-deficient host. Downregulation of Epm2a expression is widespread among mouse and human lymphoma cell lines. Epm2a-encoded laforin is a phosphatase for GSK-3beta and an important repressor in the Wnt signaling pathway. Inactivation of Epm2a resulted in increased Wnt signaling and tumorigenesis.