What a Radiologist Needs to Know About Breast Pathology Processes.

Optimal breast care requires a multidisciplinary and integrated approach, including appropriate processes and communication between the radiology and pathology departments. It is important for breast radiologists to have an understanding of the important events that occur between the time a percutaneous biopsy sample is obtained and the point at which the final pathology report is issued. This article reviews the essential processes from breast biopsy through to pathology diagnosis, including the general pathology workflow, tissue preparation, immunohistochemical staining, and pathologic reporting. Upon completion of this educational article, participants will have gained an understanding of the essential steps in the pathology workflow. This article will also highlight the important clinical information a radiologist should provide to the pathologist to ensure the most accurate and clinically relevant diagnosis. This clinical information includes the BI-RADS assessment category, the type of imaging finding that was targeted for biopsy (particularly when there are calcifications), the location of the targeted lesion relative to other findings, and other pertinent patient history.

Hit-to-lead optimization of novel phenyl imidazole carboxamides that are active against Leishmania donovani.

Visceral leishmaniasis is a potentially fatal disease caused by the parasitic protists, Leishmania donovani and L. infantum. Current treatments remain unsuitable due to cost, the need for hospitalization, variable efficacy against different species, toxicity and emerging resistance. Herein, we report the SAR exploration of the novel hit 4-Fluoro-N-(5-(4-methoxyphenyl)-1-methyl-1H-imidazole-2-yl)benzamide [1] previously identified from a high throughput screen against Trypanosoma brucei, Trypanosoma cruzi and Leishmania donovani. An extensive and informative set of analogues were synthesized incorporating key modifications around the scaffold resulting in improved potency, whilst the majority of compounds maintained low cytotoxicity against human THP-1 macrophages that are target cells for these pathogens. New lead compounds identified within this study also maintained desirable physicochemical properties, improved metabolic stability in vitro and displayed no significant mitotoxicity against HepG2 cell lines. This compound class warrants continued investigation towards development as a novel treatment for Visceral Leishmaniasis.

Unmasking the Truth of Health Care Workers' Well-being During the COVID-19 Pandemic.

BACKGROUND: Burnout is a well-documented multifactorial phenomenon that affects up to 47% of intensive care unit staff at some point in their career. The literature highlights increased rates of anxiety, depression, and posttraumatic stress disorder among staff as a result of the COVID-19 pandemic. LOCAL PROBLEM: Following the second and, at the time of writing, largest surge of the COVID-19 pandemic, concern for staff 's mental health prompted a hospital-wide study to assess depression, anxiety, posttraumatic stress disorder, and alcohol misuse and to determine the use and effectiveness of employee resources. METHODS: Through REDCap, 212 intensive care unit employees were surveyed with validated screening tools for depression, anxiety, posttraumatic stress disorder, and alcohol misuse. The use and perceived effectiveness of hospital initiatives were assessed. RESULTS: A total of 212 surveys were evaluated. Among respondents, 54% experienced mild anxiety, more than 50% screened positive for mild depression, and 37% screened positive for posttraumatic stress disorder. Most employees (74.5%) were aware of at least 1 resource; 37% knew of the COVID-19 newsletter, one of the top resources highlighted in the survey. Perceived effectiveness of resources ranged from 0.9% (disaster helpline) to 82.5% (prayers, readings, and the on-call chaplain-all "somewhat helpful"). The results correlate with the progressive trend of increased mental health concerns among intensive care unit employees. Survey results prompted an expansion of hospital resources. CONCLUSIONS: COVID-19 has brought unique mental health challenges and stressors to intensive care unit staff. By adapting and expanding resources, hospitals can improve staff resiliency and mitigate some mental health concerns with the aim of decreasing the overall psychological impact of the pandemic.

Development of mRNA manufacturing for vaccines and therapeutics: mRNA platform requirements and development of a scalable production process to support early phase clinical trials.

The remarkable success of SARS CoV-2 mRNA-based vaccines and the ensuing interest in mRNA vaccines and therapeutics have highlighted the need for a scalable clinical-enabling manufacturing process to produce such products, and robust analytical methods to demonstrate safety, potency, and purity. To date, production processes have either not been disclosed or are bench-scale in nature and cannot be readily adapted to clinical and commercial scale production. To address these needs, we have advanced an aqueous-based scalable process that is readily adaptable to GMP-compliant manufacturing, and developed the required analytical methods for product characterization, quality control release, and stability testing. We also have demonstrated the products produced at manufacturing scale under such approaches display good potency and protection in relevant animal models with mRNA products encoding both vaccine immunogens and antibodies. Finally, we discuss continued challenges in raw material identification, sourcing and supply, and the cold chain requirements for mRNA therapeutic and vaccine products. While ultimate solutions have yet to be elucidated, we discuss approaches that can be taken that are aligned with regulatory guidance.

Overcoming reduced antibiotic susceptibility in intracellular Salmonella enterica serovar Typhimurium using AR-12.

Host-directed therapies (HDTs) could enhance the activity of traditional antibiotics. AR-12 is a promising HDT against intracellular pathogens including Salmonella enterica serovar Typhimurium, and has been shown to act through modulation of autophagy and the Akt kinase pathway. Since AR-12 does not inhibit the growth of planktonic bacteria but only works in conjunction with the infected host-cell, we hypothesized that AR-12 could enhance the activity of antibiotics in less-susceptible strains in the intracellular host environment. We found that repetitive passaging of S. typhimurium in macrophages in the absence of antibiotics led to a 4-fold reduction in their intracellular susceptibility to streptomycin (STR), but had no effect on the bacteria's sensitivity to AR-12. Moreover, when the host-passaged strains were treated with a combined therapy of AR-12 and STR, there was a significant reduction of intracellular bacterial burden compared to STR monotherapy. Additionally, co-treatment of macrophages infected with multi-drug resistant S. typhimurium with AR-12 and STR or ampicillin showed enhanced clearance of the intracellular bacteria. The drug combination did not elicit this effect on planktonic bacteria. Overall, AR-12 enhanced the clearance of less susceptible S. typhimurium in an intracellular environment.

Self-esteem and self-efficacy in the status attainment process and the multigenerational transmission of advantage.

Despite considerable evidence of the importance of self-esteem and self-efficacy for agentic, goal-oriented behavior, little attention has been directed to these psychological dimensions in the status attainment literature. The present research uses data from the longitudinal, three-generation Youth Development Study (N = 422 three-generation triads) to examine the extent to which adolescent self-esteem and economic self-efficacy affect adult educational and income attainment, and whether these psychological resources are transmitted from one generation to the next, accumulating advantage across generations. We present evidence indicating that both self-esteem and economic self-efficacy are implicated in the attainment process. Adolescent economic self-efficacy had a direct positive effect on adult educational attainment and an indirect effect through educational plans. The influence of self-esteem on adult educational attainment was entirely indirect, through school achievement. We also find evidence that economic self-efficacy was transmitted from parents to children. We conclude that future research should more broadly consider psychological resources in attainment processes from a longitudinal multigenerational perspective.

Evaluation of synergy between host and pathogen-directed therapies against intracellular Leishmania donovani.

Visceral leishmaniasis (VL) is associated with treatment complications due to the continued growth of resistant parasites toward currently available pathogen-directed therapeutics. To limit the emergence and combat resistant parasites there is a need to develop new anti-leishmanial drugs and alternative treatment approaches, such as host-directed therapeutics (HDTs). Discovery of new anti-leishmanial drugs including HDTs requires suitable in vitro assay systems. Herein, we modified and evaluated a series of resazurin assays against different life-stages of the VL causing parasite, Leishmania donovani to identify novel HDTs. We further analyzed the synergy of combinatorial interactions between traditionally used pathogen-directed drugs and HDTs for clearance of intracellular L. donovani. The inhibitory concentration at 50% (IC50) of the five evaluated therapies [amphotericin B (AMB), miltefosine, paromomycin, DNER-4, and AR-12 (OSU-03012)] was determined against promastigotes, extracellular amastigotes, and intracellular amastigotes of L. donovani via a resazurin-based assay and compared to image-based microscopy. Using the resazurin-based assay, all evaluated therapies showed reproducible anti-leishmanial activity against the parasite's different life-stages. These results were consistent to the traditional image-based technique. The gold standard of therapy, AMB, showed the highest potency against intracellular L. donovani, and was further evaluated for combinatorial effects with the HDTs. Among the combinations analyzed, pathogen-directed AMB and host-directed AR-12 showed a synergistic reduction of intracellular L. donovani compared to individual treatments. The modified resazurin assay used in this study demonstrated a useful technique to measure new anti-leishmanial drugs against both intracellular and extracellular parasites. The synergistic interactions between pathogen-directed AMB and host-directed AR-12 showed a great promise to combat VL, with the potential to reduce the emergence of drug-resistant strains.

In Vivo and Cellular Trafficking of Acetalated Dextran Microparticles for Delivery of a Host-Directed Therapy for Salmonella enterica Serovar Typhi Infection.

Previously we have encapsulated host-directed therapy AR-12 into acetalated dextran (Ace-DEX) microparticles (MPs) to mitigate drug toxicity and passively target phagocytic host cells. Herein, we have improved upon our initial emulsion-based formulation of Ace-DEX MPs encapsulating AR-12 (AR-12/MPs) by improving the drug encapsulation efficiency, evaluating sterilization processes for manufacturing, and understanding cellular and in vivo trafficking of the MPs. By using an alternative solvent system, ethyl acetate, we report an increased encapsulation efficiency of AR-12 while maintaining the pH-responsive degradation kinetics of Ace-DEX MPs. To better manufacture this novel antimicrobial formulation, we sterilized AR-12/MPs by gamma irradiation or ethylene oxide and evaluated their efficacy against intracellular Salmonella enterica serovar Typhi. Sterilized AR-12/MPs resulted in a significant reduction in intracellular bacterial burden compared to Blank/MPs. We also characterized intracellular trafficking of Ace-DEX MPs encapsulating fluorophores, which demonstrated internalization of MPs in endo/lysosomal compartments and time and degradation-rate dependent lysosomal escape into cytosolic compartments. Additionally, in vivo toxicity was mitigated following encapsulation of AR-12, where the maximum tolerated dose of AR-12 was increased compared to soluble treatment via intranasal, intravenous, and intraperitoneal administration routes. Following in vivo trafficking of Ace-DEX MPs via the same routes, intranasal administration demonstrated the highest accumulation in the lungs, liver, and kidneys, which persisted out to 240 h. Overall, we have advanced the formulation of this host-directed therapy and broadened the understanding of Ace-DEX MP delivery.

Acetalated Dextran Microparticles for Codelivery of STING and TLR7/8 Agonists.

Vaccines are the most effective tool for preventing infectious diseases; however, subunit vaccines, considered the safest type, suffer from poor immunogenicity and require adjuvants to create a strong and sustained immune response. As adjuvants, pathogen-associated molecular patterns (PAMPs) offer potent immunostimulatory properties and defined mechanisms of action through their cognate pattern recognition receptors (PRRs). Their activity can be further enhanced through combining two or more PAMPs, particularly those that activate multiple immune signaling pathways. However, the cytosolic localization of many PRRs requires intracellular delivery of PAMPs for optimal biological activity, which is particularly true of the stimulator of interferon genes (STING) PRR. Using acetalated dextran (Ace-DEX) microparticles (MPs) encapsulating STING agonist 3'3'-cyclic GMP-AMP (cGAMP) combined with soluble PAMPS, we screened the effect of codelivery of adjuvants using primary mouse bone marrow derived dendritic cells (BMDCs). We identified that codelivery of cGAMP MPs and soluble Toll-like receptor 7/8 (TLR7/8) agonist resiquimod (R848) elicited the broadest cytokine response. cGAMP and R848 were then coencapsulated within Ace-DEX MPs via electrospray. Using the model antigen ovalbumin, we observed that Ace-DEX MPs coencapsulating cGAMP and R848 (cGAMP/R848 Ace-DEX MPs) induced antigen-specific cellular immunity, and a balanced Th1/Th2 humoral response that was greater than cGAMP Ace-DEX MPs alone and PAMPs delivered in separate MPs. These data indicate that polymeric Ace-DEX MPs loaded with STING and TLR7/8 agonists represent a potent cellular and humoral vaccine adjuvant.

Injectable long-acting human immunodeficiency virus antiretroviral prodrugs with improved pharmacokinetic profiles.

While highly active antiretroviral therapy (HAART) has significantly reduced mortality rates in patients with human immunodeficiency virus type 1 (HIV-1), its efficacy may be impeded by emergence of drug resistance caused by lack of patient adherence. A therapeutic strategy that requires infrequent drug administration as a result of sustained release of antiretroviral drugs would put less burden on the patient. Long-acting antiretroviral prodrugs for HIV therapy were synthesized through modification of the active drugs, emtricitabine (FTC) and elvitegravir (EVG), with docosahexaenoic acid (DHA) in one-step, one-pot, high-yielding reactions. The in vitro drug release profiles of these synthetic conjugates demonstrated sustained and controlled release of the active drug over a period of 3-4 weeks attributable to the hydrolysis of the chemical linker in conjunction with the hydrophilicity of the parent drug. Both conjugates exhibited superior antiviral activities in tissue culture models of HIV replication as compared to those of the free drugs, strengthening their role as potent prodrugs for HIV therapy. Pharmacokinetic analysis in CD1 mice further confirmed the long-acting aspect of these conjugates with released drug concentrations in plasma detected at their respective IC90/IC95 values over a period of 2 weeks and discernable amounts of active drug even at 6 weeks. Our findings suggest that the injectable small molecule conjugates could be used as long-acting controlled release of FTC and EVG in attempts to mitigate adherence-related HIV resistance.

Development of Electrospun Chitosan-Polyethylene Oxide/Fibrinogen Biocomposite for Potential Wound Healing Applications.

Normal wound healing is a highly complex process that requires the interplay of various growth factors and cell types. Despite advancements in biomaterials, only a few bioactive wound dressings reach the clinical setting. The purpose of this research was to explore the feasibility of electrospinning a novel nanofibrous chitosan (CS)-fibrinogen (Fb) scaffold capable of sustained release of platelet-derived growth factor (PDGF) for the promotion of fibroblast migration and wound healing. CS-Fb scaffolds were successfully electrospun using a dual-spinneret electrospinner and directly evaluated for their physical, chemical, and biological characteristics. CS-polyethylene/Fb scaffolds exhibited thinner fiber diameters than nanofibers electrospun from the individual components while demonstrating adequate mechanical properties and homogeneous polymer distribution. In addition, the scaffold demonstrated acceptable water transfer rates for wound healing applications. PDGF was successfully incorporated in the scaffold and maintained functional activity throughout the electrospinning process. Furthermore, released PDGF was effective at promoting fibroblast migration equivalent to a single 50 ng/mL dose of PDGF. The current study demonstrates that PDGF-loaded CS-Fb nanofibrous scaffolds possess characteristics that would be highly beneficial as novel bioactive dressings for enhancement of wound healing.

Genomic and transcriptomic comparison of allergen and silver nanoparticle-induced mast cell degranulation reveals novel non-immunoglobulin E mediated mechanisms.

Mast cells represent a crucial cell type in host defense; however, maladaptive responses are contributing factors in the pathogenesis of allergic diseases. Previous work in our laboratory has shown that exposure to silver nanoparticles (AgNPs) results in mast cell degranulation via a non-immunoglobulin E (IgE) mechanism. In this study, we utilized a systems biology approach to identify novel genetic factors playing a role in AgNP-induced mast cell degranulation compared to the classical activation by antigen-mediated FcεRI crosslinking. Mast cell degranulation was assessed in bone marrow-derived mast cells isolated from 23 strains of mice following exposure to AgNPs or FcεRI crosslinking with dinitrophenyl (DNP). Utilizing strain-dependent mast cell degranulation, an association mapping study identified 3 chromosomal regions that were significantly associated with mast cell degranulation by AgNP and one non-overlapping region associated with DNP-mediated degranulation. Two of the AgNP-associated regions correspond to genes previously reported to be associated with allergic disorders (Trac2 on chromosome 1 and Traf6 on chromosome 2) and an uncharacterized gene identified on chromosome 1 (Fam126b). In conjunction, RNA-sequencing performed on mast cells from the high and low responder strains revealed 3754 and 34 differentially expressed genes that were unique to DNP and AgNP exposures, respectively. Select candidate genes include Ptger4, a gene encoding a G-protein coupled receptor in addition to a multifunctional adaptor protein, Txnip, that may be driving mast cell degranulation by AgNP. Taken together, we identified novel genes that have not been previously shown to play a role in nanoparticle-mediated mast cell activation. With further functional evaluation in the future, these genes may be potential therapeutic targets in the treatment of non-IgE mediated mast cell-linked disorders.

Tunable degradation of acetalated dextran microparticles enables controlled vaccine adjuvant and antigen delivery to modulate adaptive immune responses.

Subunit vaccines are often poorly immunogenic, and adjuvants and/or delivery vehicles, such as polymeric microparticles (MPs), can be used to enhance immune responses. MPs can also be used to understand cell activation kinetics and the significant impact antigen and adjuvant release has on adaptive immune responses. By controlling antigen and adjuvant release, we can determine if it is important to have precise temporal control over release of these elements to optimize the peak and duration of protective immunity and improve vaccine safety profiles. In order to study the effect of tunable adjuvant or antigen delivery on generation of adaptive immunity, we used acetalated dextran (Ace-DEX) MPs. Ace-DEX MPs were used because their tunable degradation can be controlled based on polymer cyclic acetal coverage (CAC). Ace-DEX MPs of varying degradation profiles were used to deliver murabutide or ovalbumin (OVA) as a model adjuvant or antigen, respectively. When murabutide was encapsulated within Ace-DEX MPs to test for controlled adjuvant delivery, fast-degrading MPs exhibited higher humoral and cellular responses in vivo at earlier time points, while slow-degrading MPs resulted in stronger responses at later time points. When OVA was encapsulated within Ace-DEX MPs to test for controlled antigen delivery, fast-degrading MPs induced greater antibody and cytokine production throughout the length of the experiment. This differential response suggests the need for distinct, flexible control over adjuvant or antigen delivery and its impact on immune response modulation.

Vaccines for the Prevention of Melioidosis and Glanders.

PURPOSE OF REVIEW: Burkholderia pseudomallei's and Burkholderia mallei's high rate of infectivity, limited treatment options, and potential use as biological warfare agents underscore the need for development of effective vaccines against these bacteria. Research efforts focused on vaccines against these bacteria are in pre-clinical stages, with no approved formulations currently on the market. RECENT FINDINGS: Several live attenuated and subunit vaccine formulations have been evaluated in animal studies, with no reports of significant long term survival after lethal challenge. SUMMARY: This review encompasses the most current vaccine strategies to prevent B. pseudomallei and B. mallei infections while providing insight for successful vaccines moving forward.

Adolescent Agentic Orientations: Contemporaneous Family Influence, Parental Biography and Intergenerational Development.

Agentic orientations developed in adolescence have been linked to better health, well-being, and achievements in the years following. This study examines longitudinal parental influences on the development of adolescent children's agentic orientations, captured by the core constructs of mastery beliefs and generalized life expectations. Drawing on multigenerational panel data from the United States (1991-2011), the study examines contemporaneous family factors, but also how parental biographies (their own transition to adulthood) and parents' own adolescent agentic orientations influence their adolescent children. Study adolescents were 46% male, 52% white, and 15.6 years old on average. The findings indicate that parents' early orientations and experiences in the transition to adulthood have little effect on their children's mastery beliefs, but that parents' generalized life expectations (in adolescence) and having married before having the child were associated with their children's more optimistic life expectations. Contemporaneous family income and optimistic expectations among parents-as-adolescents were somewhat substitutable as positive influences on adolescents' optimistic life expectations. The findings contribute to our understanding of intergenerational and over-time influences on these key adolescent orientations.

Familial Transmission of Educational Plans and the Academic Self-Concept: A Three-Generation Longitudinal Study.

This research investigates the social reproduction of inequality by drawing on prospective longitudinal data from three generations of Youth Development Study respondents. It examines intergenerational influence on the relatively unexplored academic self-concept as well as educational plans, a critical component of the status attainment model. A structural equation model, based on 422 3-generation triads, finds evidence that the sources giving rise to the development of children's (Generation 3) achievement orientations do not only result from parental (G2) contemporaneous influence. Prior influences implicate grandparent (G1) educational attainment and income, grandparental expectations for the G2 adolescent, the G2 academic self-concept and educational plans measured more than twenty years earlier (in G2's adolescence), and G2 educational attainment. A familial culture emphasizing academic self-confidence and high educational expectations may be an important component of "family capital" that supports educational attainment and contributes to the maintenance of social class position in each successive generation.

Family (Dis)Advantage and Life Course Expectations.

Optimistic assessments of life chances can positively influence life outcomes, but conflicting theories suggest these assessments either reflect structural privilege or develop as a result of childhood hardship. In addition, competing hypotheses suggest that these assessments may matter differently depending on who holds them. We examine whether family socioeconomic status shapes adolescents' expectations about how successful their lives will turn out. We distinguish generalized life expectations (GLE), capturing anticipated success in life across multiple domains, from intergenerational comparative expectations (ICE), which register expectations about improvement relative to observed success within the respondent's family lineage. We find that adolescents from higher socioeconomic status families are simultaneously more optimistic about their likely success in life (GLE) but less likely to anticipate relative improvement in life success across generations (ICE). Holding high GLE in combination with low ICE predicted doing better in adulthood across a range of health, attainment, and well-being outcomes, though in most cases high GLE, regardless of ICE, was the key. These beneficial patterns are, for the most part, at least as beneficial for socioeconomically disadvantaged youth as they are for advantaged youth.

Contribution of engineered nanomaterials physicochemical properties to mast cell degranulation.

The rapid development of engineered nanomaterials (ENMs) has grown dramatically in the last decade, with increased use in consumer products, industrial materials, and nanomedicines. However, due to increased manufacturing, there is concern that human and environmental exposures may lead to adverse immune outcomes. Mast cells, central to the innate immune response, are one of the earliest sensors of environmental insult and have been shown to play a role in ENM-mediated immune responses. Our laboratory previously determined that mast cells are activated via a non-FcεRI mediated response following silver nanoparticle (Ag NP) exposure, which was dependent upon key physicochemical properties. Using bone marrow-derived mast cells (BMMCs), we tested the hypothesis that ENM physicochemical properties influence mast cell degranulation. Exposure to 13 physicochemically distinct ENMs caused a range of mast degranulation responses, with smaller sized Ag NPs (5 nm and 20 nm) causing the most dramatic response. Mast cell responses were dependent on ENMs physicochemical properties such as size, apparent surface area, and zeta potential. Surprisingly, minimal ENM cellular association by mast cells was not correlated with mast cell degranulation. This study suggests that a subset of ENMs may elicit an allergic response and contribute to the exacerbation of allergic diseases.

Adolescent adaptation before, during and in the aftermath of the Great Recession in the USA.

This study examines the impact of the "Great Recession" (from December 2007 to June 2009) on 8th and 10th graders in the USA, using annual nationally representative data from the Monitoring the Future study. Historical changes in youth adjustment (self-esteem, depressed mood, risk taking, aggression and property crime), school achievement (grade point average [GPA], time spent on homework and educational expectations) and structured and unstructured activities (volunteering, employment, sports and evenings out for fun) were examined between 1991 and 2014. Overall, there were only slight changes in mean levels of adjustment, achievement and most youth activities. However, the percentage of youth working during the school year did decline during the Great Recession. Several longer-term trends were also evident, though not directly tied to the Great Recession. These include an increase in GPA, a decrease in time spent on homework, rising educational expectations and more time spent volunteering. Future work should assess how the shift to unpaid work activities (e.g. volunteering and internships) among youth is impacting the transition from school to work in the contemporary economy, and whether the Great Recession had deleterious impacts for younger children or among youth whose parents lost work or had their homes foreclosed.