Pathogenic diversification of the gut commensal Providencia alcalifaciens via acquisition of a second type III secretion system.

Providencia alcalifaciens is a Gram-negative bacterium found in various water and land environments and organisms, including insects and mammals. Some P. alcalifaciens strains encode gene homologs of virulence factors found in pathogenic Enterobacterales members, such as Salmonella enterica serovar Typhimurium and Shigella flexneri. Whether these genes are pathogenic determinants in P. alcalifaciens is not known. In this study, we investigated P. alcalifaciens-host interactions at the cellular level, focusing on the role of two type III secretion systems (T3SS) belonging to the Inv-Mxi/Spa family. T3SS1b is widespread in Providencia spp. and encoded on the chromosome. A large plasmid that is present in a subset of P. alcalifaciens strains, primarily isolated from diarrheal patients, encodes for T3SS1a. We show that P. alcalifaciens 205/92 is internalized into eukaryotic cells, lyses its internalization vacuole, and proliferates in the cytosol. This triggers caspase-4-dependent inflammasome responses in gut epithelial cells. The requirement for the T3SS1a in entry, vacuole lysis, and cytosolic proliferation is host cell type-specific, playing a more prominent role in intestinal epithelial cells than in macrophages or insect cells. In a bovine ligated intestinal loop model, P. alcalifaciens colonizes the intestinal mucosa and induces mild epithelial damage with negligible fluid accumulation in a T3SS1a- and T3SS1b-independent manner. However, T3SS1b was required for the rapid killing of Drosophila melanogaster. We propose that the acquisition of two T3SS has allowed P. alcalifaciens to diversify its host range, from a highly virulent pathogen of insects to an opportunistic gastrointestinal pathogen of animals.

A Scoping Review of Organ Transplantation in Populations Experiencing Incarceration.

Despite an aging confined population, the current state of organ transplantation in carceral systems is largely unknown. This scoping review aimed to assess the literature on organ transplantation in populations experiencing incarceration. The review used the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for a scoping review. Included references were published between January 2000 and January 2022 in PubMed, Cumulative Index to Nursing and Allied Health Literature via EBSCO, EMBASE.com, PsycInfo via EBSCO, Sociological Abstracts via ProQuest, and Scopus. Two reviewers conducted title and abstract screening, full-text review, and data extraction in order to generate common themes. The initial search yielded 3,225 studies, and 2,129 references underwent screening. Seventy studies underwent full-text review, and 10 met inclusion criteria. These studies revealed heterogeneous perspectives and policies by providers and transplant centers regarding transplant consideration of individuals with incarceration history or current involvement. Two studies on a kidney transplant program for patients experiencing incarceration showed transplant as a sustainable and potentially superior option for people who are incarcerated, as compared with chronic hemodialysis. Literature on transplantation for populations experiencing incarceration is sparse. More research is required to understand the demand for transplants and the ethical implications of the heterogeneous perspectives and policies on practice patterns.

Pathogenic diversification of the gut commensal Providencia alcalifaciens via acquisition of a second type III secretion system.

Providencia alcalifaciens is a Gram-negative bacterium found in a wide variety of water and land environments and organisms. It has been isolated as part of the gut microbiome of animals and insects, as well as from stool samples of patients with diarrhea. Specific P. alcalifaciens strains encode gene homologs of virulence factors found in other pathogenic members of the same Enterobacterales order, such as Salmonella enterica serovar Typhimurium and Shigella flexneri. Whether these genes are also pathogenic determinants in P. alcalifaciens is not known. Here we have used P. alcalifaciens 205/92, a clinical isolate, with in vitro and in vivo infection models to investigate P. alcalifaciens -host interactions at the cellular level. Our particular focus was the role of two type III secretion systems (T3SS) belonging to the Inv-Mxi/Spa family. T3SS 1b is widespread in Providencia spp. and encoded on the chromosome. T3SS 1a is encoded on a large plasmid that is present in a subset of P. alcalifaciens strains, which are primarily isolates from diarrheal patients. Using a combination of electron and fluorescence microscopy and gentamicin protection assays we show that P. alcalifaciens 205/92 is internalized into eukaryotic cells, rapidly lyses its internalization vacuole and proliferates in the cytosol. This triggers caspase-4 dependent inflammasome responses in gut epithelial cells. The requirement for the T3SS 1a in entry, vacuole lysis and cytosolic proliferation is host-cell type specific, playing a more prominent role in human intestinal epithelial cells as compared to macrophages. In a bovine ligated intestinal loop model, P. alcalifaciens colonizes the intestinal mucosa, inducing mild epithelial damage with negligible fluid accumulation. No overt role for T3SS 1a or T3SS 1b was seen in the calf infection model. However, T3SS 1b was required for the rapid killing of Drosophila melanogaster . We propose that the acquisition of two T3SS by horizontal gene transfer has allowed P. alcalifaciens to diversify its host range, from a highly virulent pathogen of insects to an opportunistic gastrointestinal pathogen of animals.

Nonsignaling extracellular spacer regulates tumor antigen selectivity of CAR T cells.

Advancing chimeric antigen receptor (CAR)-engineered T cells for the treatment of solid tumors is a major focus in the field of cellular immunotherapy. Several hurdles have hindered similar CAR T cell clinical responses in solid tumors as seen in hematological malignancies. These challenges include on-target off-tumor toxicities, which have inspired efforts to optimize CARs for improved tumor antigen selectivity and overall safety. We recently developed a CAR T cell therapy targeting prostate stem cell antigen (PSCA) for prostate and pancreatic cancers, showing improved preclinical antitumor activity and T cell persistence by optimizing the intracellular co-stimulatory domain. Similar studies were undertaken to optimize HER2-directed CAR T cells with modifications to the intracellular co-stimulatory domain for selective targeting of breast cancer brain metastasis. In the present study, we evaluate various nonsignaling extracellular spacers in these CARs to further improve tumor antigen selectivity. Our findings suggest that length and structure of the extracellular spacer can dictate the ability of CARs to selectively target tumor cells with high antigen density, while sparing cells with low antigen density. This study contributes to CAR construct design considerations and expands our knowledge of tuning solid tumor CAR T cell therapies for improved safety and efficacy.

State of the Interactomes: an evaluation of molecular networks for generating biological insights.

Advancements in genomic and proteomic technologies have powered the use of gene and protein networks ("interactomes") for understanding genotype-phenotype translation. However, the proliferation of interactomes complicates the selection of networks for specific applications. Here, we present a comprehensive evaluation of 46 current human interactomes, encompassing protein-protein interactions as well as gene regulatory, signaling, colocalization, and genetic interaction networks. Our analysis shows that large composite networks such as HumanNet, STRING, and FunCoup are most effective for identifying disease genes, while smaller networks such as DIP and SIGNOR demonstrate strong interaction prediction performance. These findings provide a benchmark for interactomes across diverse network biology applications and clarify factors that influence network performance. Furthermore, our evaluation pipeline paves the way for continued assessment of emerging and updated interaction networks in the future.

Investigating the effect of anatomical variations in the response of the neonatal brachial plexus to applied force: Use of a two-dimensional finite element model.

The brachial plexus is a set of nerves that innervate the upper extremity and may become injured during the birthing process through an injury known as Neonatal Brachial Plexus Palsy. Studying the mechanisms of these injuries on infant cadavers is challenging due to the justifiable sensitivity surrounding testing. Thus, these specimens are generally unavailable to be used to investigate variations in brachial plexus injury mechanisms. Finite Element Models are an alternative way to investigate the response of the neonatal brachial plexus to loading. Finite Element Models allow a virtual representation of the neonatal brachial plexus to be developed and analyzed with dimensions and mechanical properties determined from experimental studies. Using ABAQUS software, a two-dimensional brachial plexus model was created to analyze how stresses and strains develop within the brachial plexus. The main objectives of this study were (1) to develop a model of the brachial plexus and validate it against previous literature, and (2) to analyze the effect of stress on the nerve roots based on variations in the angles between the nerve roots and the spinal cord. The predicted stress for C5 and C6 was calculated as 0.246 MPa and 0.250 MPa, respectively. C5 and C6 nerve roots experience the highest stress and the largest displacement in comparison to the lower nerve roots, which correlates with clinical patterns of injury. Even small (+/- 3 and 6 degrees) variations in nerve root angle significantly impacted the stress at the proximal nerve root. This model is the first step towards developing a complete three-dimensional model of the neonatal brachial plexus to provide the opportunity to more accurately assess the effect of the birth process on the stretch within the brachial plexus and the impact of biological variations in structure and properties on the risk of Neonatal Brachial Plexus Palsy.

Measuring Physical Function in the PICU: Development and Testing of a Children's Version (Age 2-18 yr) of the Chelsea Critical Care Physical Assessment Tool.

OBJECTIVES: To adapt and develop a reliable and easily administered outcome measure of physical and respiratory function in critically ill children in the PICU. DESIGN: Modified Delphi study to adapt the Chelsea Critical Care Physical Assessment (CPAx) tool for use in children 2-18 years old, with subsequent prospective testing in a single-center cohort. SETTING: Single-center tertiary PICU. SUBJECTS: Delphi process in 27 panelists (including physiotherapists, occupational therapists, and pediatric intensivists from seven countries from January 2018 to March 2018). Cohort study in 54 patients admitted to PICU for greater than 24 hours over a 3-month period (April 2018 to June 2018), with median age 5.5 years (interquartile range [IQR], 3-12.75 yr), 33 of 54 male, and 38 of 54 invasively ventilated. INTERVENTIONS: None. MEASUREMENTS AND MAIN RESULTS: Three Delphi iterations were required to reach greater than or equal to 80% consensus in all the children's CPAx (cCPAx) items. In the subsequent cohort study, six physiotherapists used the cCPAx tool and scored 54 participants, with a total 106 observations. The median cCPAx tool score was 14.50 (IQR, 3-25) out of a possible total of 50. Inter-rater reliability for 30 randomly selected participants was excellent (intraclass correlation coefficient, 0.998). Completion rate of cCPAx in the 54 patients occurred in 78 of 106 occasions (74%). CONCLUSIONS: The cCPAx tool content that was developed using Delphi methodology provided a feasible and clinically relevant tool for use in assessing physical morbidity in PICU patients 2-18 years old. Overall, the cCPAx scores were low, demonstrating low levels of physical function and high levels of immobility during PICU care.

What do measures of gender identity tell us about gender identity over time?

Gender identity is a multifaceted concept and is represented by a wide range of measures and constructs including both self-report and researcher observations of preferences and behaviours. However, despite their similar theoretical underpinning, gender identity measures are rarely found to correlate with one another, and contrasting patterns and trajectories are often found for each construct (Egan & Perry, Developmental Psychology, 37, 2001, 451). Therefore, this systematic review aimed to present a review of the longitudinal research evidence surrounding gender identity development in the absence of formal intervention. Using a systematic search strategy, 21 studies were identified. Narrative synthesis was used to synthesize the data collected in these studies and trajectories were explored for (1) self-identification measures of gender identity, (2) clothing preferences, (3) peer preferences, and (4) object/activity preferences. Overall, the results of this systematic review are consistent with wider research suggesting that distinct developmental patterns can be observed when using different constructs and measures of gender identity.

Crystal size, shape, and conformational changes drive both the disappearance and reappearance of ritonavir polymorphs in the mill.

Organic compounds can crystallize in different forms known as polymorphs. Discovery and control of polymorphism is crucial to the pharmaceutical industry since different polymorphs can have significantly different physical properties which impacts their utilization in drug delivery. Certain polymorphs have been reported to 'disappear' from the physical world, irreversibly converting to new ones. These unwanted polymorph conversions, initially prevented by slow nucleation kinetics, are eventually observed driven by significant gains in thermodynamic stabilities. The most infamous of these cases is that of the HIV drug ritonavir (RVR): Once its reluctant form was unwillingly nucleated for the first time, its desired form could no longer be produced with the same manufacturing process. Here we show that RVR's extraordinary disappearing polymorph as well as its reluctant form can be consistently produced by ball-milling under different environmental conditions. We demonstrate that the significant difference in stability between its polymorphs can be changed and reversed in the mill-a process we show is driven by crystal size as well as crystal shape and conformational effects. We also show that those effects can be controlled through careful design of milling conditions since they dictate the kinetics of crystal breakage, dissolution, and growth processes that eventually lead to steady-state crystal sizes and shapes in the mill. This work highlights the huge potential of mechanochemistry in polymorph discovery of forms initially difficult to nucleate, recovery of disappearing polymorphs, and polymorph control of complex flexible drug compounds such as RVR.

Test Anxiety Among US Medical Students: A Review of the Current Literature.

Purpose: Medical students experience anxiety at higher rates than the general public and many are uniquely affected by additional test anxiety throughout their medical education. Although test anxiety has been studied for decades, little evidence has been published suggesting interventions improve examination performance in medical education. Therefore, we set out to review the current literature to elucidate efforts so far and establish trends in research. Methods: Databases searched included PubMed, EMBASE, PsychINFO, ERIC, SCOPUS, and CINAHL. English language articles published between 2010 and 2021 were loaded into a reference manager to screen out duplicate articles. During the full-text screen and data extraction phase, reference lists were also inspected to identify additional articles for inclusion in the study. Results: Of 883 studies identified, 860 were excluded resulting in 22 studies for extraction and analysis. First-year (n = 15) and second-year (n = 12) students were primarily tested. Less than 10 included third- or fourth-year students. Self-help and wellness interventions were employed, though interventions ranged from dog therapy to deep breathing techniques to fish oil supplementation. Test anxiety was evaluated using self-report questionnaires, such as the Westside Test Anxiety Scale, Beck Anxiety Inventory, and State-Trait Anxiety Inventory. None of the studies reported improved examination scores. Conclusion: This review identifies a variety of measurement tools and interventions attempting to mitigate test anxiety. As far as improving examination performance, none of the interventions reported was successful. Further research addressing test anxiety that results in improved medical student academic performance should be conducted and also use established assessment tools.

Review of neoplasia in fish at a large display aquarium, 2005-2021.

Fish maintained in managed care may have longer lifespans as a result of advances in veterinary medicine and husbandry and reduced risk of predation. Neoplasia is of increasing interest in managed aquarium populations. However, few studies have systematically evaluated neoplasia in managed fish populations. Our objective in this retrospective study was to review and describe neoplasia diagnosed in fish at a large public display aquarium between 2005 and 2021. Any fish diagnosed with neoplasia on either antemortem or postmortem evaluation during the study period was included, and all medical records, biopsy, and autopsy reports were reviewed. Sixty-two fish met the inclusion criteria; 37 species were included in the study population, most of which were tropical freshwater fish (n = 34 fish). Thirty-two types of neoplasia were identified. Ten fish had benign neoplasms, and 53 fish had malignant neoplasms. The most common neoplasms were of epithelial and neuroectodermal origin. The most common site of tumor origin was the skin. Our data suggest that mesenchymal neoplasms may be more common in cold saltwater fish than in tropical freshwater and saltwater fish. Malignant neoplasms were most commonly diagnosed in the study population and should be a top differential when neoplasms are identified in fish managed under human care. Our study contributes to the overall knowledge of the health of aquarium fish and may aid clinicians in characterizing neoplasia that may be present in fish under human care.

Navigating the landscape: Roles, perspectives, and experiences of psychologists supporting gender diverse children and young people in school settings.

Purpose: This review aimed to explore and synthesize the perspectives and experiences of school-based psychological professionals providing support to gender diverse CYP across the world, to foreground the voices of those with relevant experience and support future practice. Methods: A systematic review of five databases (PsychINFO, CINAHL, ERIC, SCOPUS and PROQUEST dissertations and theses) was performed between September and November 2022. Articles were included if they contained qualitative, primary research data representing the voice of at least one school-based psychological professional with experience working with gender diverse children and young people. Articles were excluded if they did not contain primary research data, were quantitative, related to non-school based psychologists or focussed on participant views in the absence of direct experience working with gender diverse pupils. Articles were thematically summarized and organized into a data extraction table. Results: Eighteen studies were identified for review, including 11 studies based in the USA, five in the UK, one in Australia and one in Cyprus. The voices of school-based professionals, including school counselors, school psychologists, trainee and qualified educational psychologists, were represented. The themes created highlighted the importance of the environment in which psychologists were working, the reliance on their own views and values to guide their work in the absence of clear guidance, the role psychologists saw they had to advocate for gender diverse CYP, as well as barriers and systems they were fighting against. Conclusion: The review highlighted the need for psychologists to advocate for gender diverse children and young people, in an often non-inclusive environment where there was a need to work systemically with schools. Future research is needed to explore young people's experiences of the support that they are receiving and would like to receive.

A game of raids: Expanding on a game theoretical approach utilising the prisoner's dilemma and ethnography in situ.

In this commentary, we set out the specifics of how Glowacki's game theoretical framework for the evolution of peace could be incorporated within broader cultural evolutionary approaches. We outline a formal proposal for prisoner's dilemma games investigating raid-based conflict. We also centre an ethnographic lens to understand the norms surrounding war and peace in intergroup interactions in small-scale communities.

Research Review: A systematic review and meta-analysis of sex differences in narrow constructs of restricted and repetitive behaviours and interests in autistic children, adolescents, and adults.

BACKGROUND: Evidence that autism often manifests differently between males and females is growing, particularly in terms of social interaction and communication, but it is unclear if there are sex differences in restricted and repetitive behaviours and interests (RRBIs) when rigorously focusing on the narrow construct level (i.e., stereotyped behaviour, restricted interests, insistence on sameness, and/or sensory experiences). METHODS: We conducted a systematic review and four random effects meta-analyses investigating sex differences in narrow construct measures of RRBIs in autistic children, adolescents, and adults (Prospero registration ID: CRD42021254221). Study quality was appraised using the Newcastle-Ottawa Quality Assessment Scale. RESULTS: Forty-six studies were narratively synthesised and 25 of these were included in four random effects meta-analyses. Results found that autistic males had significantly higher levels of stereotyped behaviours (SMD = 0.21, 95% confidence interval (CI) [0.09, 0.33], p < .001) and restricted interests (SMD = 0.18, 95% CI [0.07, 0.29], p < .001) compared to autistic females. In contrast, there were no significant sex differences for sensory experiences (SMD = -0.09, 95% CI [-0.27, 0.09], p = .32) and insistence on sameness (SMD = 0.01, 95% CI [-0.03, 0.05], p = .68). The findings from the narrative synthesis were generally consistent with those from the meta-analyses and also found qualitative sex differences in the way RRBIs manifest. CONCLUSIONS: Our findings show significant differences in narrowly defined RRBIs in males and females. Practitioners need to be aware of such differences, which could be contributing to the under-recognition of autism in females and may not be captured by current diagnostic instruments.

Understanding the value of curation: A survey of researcher perspectives of data curation services from six US institutions.

Data curation encompasses a range of actions undertaken to ensure that research data are fit for purpose and available for discovery and reuse, and can help to improve the likelihood that data is more FAIR (Findable, Accessible, Interoperable, and Reusable). The Data Curation Network (DCN) has taken a collaborative approach to data curation, sharing curation expertise across a network of partner institutions and data repositories, and enabling those member institutions to provide expert curation for a wide variety of data types and discipline-specific datasets. This study sought to assess the satisfaction of researchers who had received data curation services, and to learn more about what curation actions were most valued by researchers. By surveying researchers who had deposited data into one of six academic generalist data repositories between 2019-2021, this study set out to collect feedback on the value of curation from the researchers themselves. A total of 568 researchers were surveyed; 42% (238) responded. Respondents were positive in their evaluation of the importance and value of curation, indicating that the participants not only value curation services, but are largely satisfied with the services provided. An overwhelming majority 97% of researchers agreed that data curation adds value to the data sharing process, 96% agreed it was worth the effort, and 90% felt more confident sharing their data due to the curation process. We share these results to provide insights into researchers' perceptions and experience of data curation, and to contribute evidence of the positive impact of curation on repository depositors. From the perspective of researchers we surveyed, curation is worth the effort, increases their comfort with data sharing, and makes data more findable, accessible, interoperable, and reusable.

Breast Cancer Germline Genetic Counseling and Testing for Populations of African Heritage Globally: A Scoping Review on Research, Practice, and Bioethical Considerations.

PURPOSE: Despite the disproportionately high risk of breast cancer among women of African heritage, little is known about the facilitators and barriers to implementing germline genetic testing and counseling (GT/C). METHODS: This scoping review followed guidelines recommended by the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for scoping reviews. Published manuscripts from database inception through 2021 were sourced from PubMed, Cumulative Index to Nursing and Allied Health Literature via EBSCO, Embase, Cochrane Library, and Scopus. Search terms were used to retrieve articles addressing (1) African heritage, (2) breast cancer, and (3) GT or GC. The screening involved abstract and title review and full-text review. Data were extracted for all articles meeting the inclusion criteria. RESULTS: A total of 154 studies were included. Most studies that took place were conducted in the United States (71.4%), and most first authors (76.9%) were from the United States. GT was conducted in 73 (49.7%) studies. BRCA1/BRCA2 were the most commonly studied genes for germline mutations. GC was conducted in 49 studies (33.3%), and perspectives on GC were evaluated in 43 (29.3%). The use of racial/ethnic categories varied broadly, although African American was most common (40.1%). Racism was mentioned in three studies (2.0%). CONCLUSION: There is a growing body of literature on GT/C for breast cancer in women of African heritage. Future studies on GT/C of African populations should consider increased clarity around racial/ethnic categorizations, continued community engagement, and intentional processes for informed consent.

Techniques and Fixation of Olecranon Osteotomy: A Systematic Review.

Purpose: We aimed to characterize the incidence of complications regarding olecranon osteotomy, looking more specifically at the type of osteotomy and the fixation construct used to repair the osteotomy. Methods: In accordance with Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines, a comprehensive search was performed. A study was included if it was an adult clinical study, a transverse or chevron olecranon osteotomy was performed, and the study explicitly states the fixation construct used to repair the osteotomy. A quality assessment was performed in each study prior to data extraction. Results: We included 39 studies with a total of 1,445 patients. Most studies included patients who were being treated primarily for a distal humerus fracture. The overall incidence of delayed union was 27/643 (4.2%), with a higher rate in transverse osteotomy than in chevron osteotomy (5/49 (10.2%) vs 22/595 (3.7%)). Nonunion occurred in 43/811 (5.4%) of patients, with a higher rate in transverse osteotomy (6/73 (8.2%) vs. 37/712 (5.2%)). Implant failure or loss of reduction occurred in 44/746 (5.9%) of patients, with a higher rate in transverse osteotomy (11/49 (22.4%) vs 33/688 (4.8%)). The removal of implants occurred in 236/1078 (21.9%) of all patients, with the highest rate in those studies that used plate fixation 44/99 (44.4%). Conclusions: Compared with chevron osteotomy, patients who underwent transverse osteotomy had a higher incidence of delayed union, nonunion, and implant failure or loss of reduction requiring revision surgery. The incidence of implant removal indicates that patients should be informed that nearly half of the osteotomy fixed with a plate was removed after implantation. Type of study/level of evidence: Therapeutic III.