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Abstract BamA is the central component of the essential β-barrel assembly machine (BAM), a conserved multi-subunit complex that dynamically inserts and folds β-barrel proteins into the outer membrane of Gram-negative bacteria. Despite recent advances in our mechanistic and structural understanding of BamA, there are few potent and selective tool molecules that can bind to and modulate BamA activity. Here, we explored in vitro selection methods and different BamA/BAM protein formulations to discover peptide macrocycles that kill Escherichia coli by targeting extreme conformational states of BamA. Our studies show that Peptide Targeting BamA-1 (PTB1) targets an extracellular divalent cation-dependent binding site and locks BamA into a closed lateral gate conformation. By contrast, PTB2 targets a luminal binding site and traps BamA into an open lateral gate conformation. Our results will inform future antibiotic discovery efforts targeting BamA and provide a template to prospectively discover modulators of other dynamic integral membrane proteins.

Abstract Amniotes feature two principal visual processing systems: the tectofugal and thalamofugal pathways. In most mammals, the thalamofugal pathway predominates, routing retinal afferents through the dorsolateral geniculate complex to the visual cortex. In most birds, the thalamofugal pathway often plays the lesser role with retinal afferents projecting to the principal optic thalami, a complex of several nuclei that resides in the dorsal thalamus. This thalamic complex sends projections to a forebrain structure called the Wulst, the terminus of the thalamofugal visual system. The thalamofugal pathway in birds serves many functions such as pattern discrimination, spatial memory, and navigation/migration. A comprehensive analysis of avian species has unveiled diverse subdivisions within the thalamic and forebrain structures, contingent on species, age, and techniques utilized. In this study, we documented the thalamofugal system in three dimensions by integrating histological and contrast-enhanced computed tomography imaging of the avian brain. Sections of two-week-old chick brains were cut in either coronal, sagittal, or horizontal planes and stained with Nissl and either Gallyas silver or Luxol Fast Blue. The thalamic principal optic complex and pallial Wulst were subdivided on the basis of cell and fiber density. Additionally, we utilized the technique of diffusible iodine-based contrast-enhanced computed tomography (diceCT) on a 5-week-old chick brain, and right eyeball. By merging diceCT data, stained histological sections, and information from the existing literature, a comprehensive three-dimensional model of the avian thalamofugal pathway was constructed. The use of a 3D model provides a clearer understanding of the structural and spatial organization of the thalamofugal system. The ability to integrate histochemical sections with diceCT 3D modeling is critical to better understanding the anatomical and physiologic organization of complex pathways such as the thalamofugal visual system.

Fugitive emissions of methane (CH4) from force main sewers are of increasing concern. Dosing of additives into force main sewers could be employed to mitigate methane emissions. However, all additives will have embodied greenhouse gas (GHG) emissions. This study examined commonly employed additives in terms of modes of action and potential to mitigate methane generation. Typical dosing strategies reported in the literature for each chemical were compiled and their embodied GHG emissions were summarised from sources in the literature. The net emissions considering mitigated methane generation and embodied GHG emissions were calculated on the basis of typical usage reported in the literature. The results revealed that biofilm shocking strategies and addition of iron have the greatest net reduction in GHG emissions. There is, however, uncertainty associated with the mechanisms by which iron reduces CH4 generation in force mains. Furthermore, future changes in the sourcing of iron may increase its embodied emissions. A qualitative assessment of the impacts of additive use on downstream GHG emissions revealed that they are highly case specific. HIGHLIGHTS Sewer additives for reducing CH4 generation are compared.; Embodied emissions of additives are estimated.; Estimates of embodied emissions with reductions in CH4 generation are compared.; Additives with the best net emissions are identified.;

In this state-of-the-art review we detail the journey of xenotransplantation from its infancy, detailing one of the first published cases and the subsequent journey the field took in its inception and development. With a focus on the science, technological advances, precautions required along with the potential limitations in application, the ethics, guidance’s, and legislative advances that are required to reach the safe and efficacious clinical application of xenotransplantation. Along with a view over the past several decades with the overall significant advancements in pre-clinical study outcomes particularly in islet, kidney, and heart xenotransplantation, to ultimately reach the pinnacle of successful clinical heart and kidney xenotransplants. It outlines the importance for the appropriate guidance’s required to have been developed by experts, scientists, clinicians, and other players who helped develop the field over the past decades. It also touches upon patient advocacy along with perspectives and expectations of patients, along with public opinion and media influence on the understanding and perception of xenotransplantation. It discusses the legislative environment in different jurisdictions which are reviewed in line with current clinical practices. All of which are ultimately based upon the guidance’s developed from a strong long-term collaboration between the International Xenotransplantation Association, the World Health Organisation and The Transplantation Society; each having constantly undertaken consultation and outreach to help develop best practice for clinical xenotransplantation application. These clearly helped forge the legislative frameworks required along with harmonization and standardization of regulations which are detailed here. Also, in relation to the significant advances in the context of initial xeno-kidney trials and the even greater potential for clinical xeno-islet trials to commence we discuss the significant advantages of xenotransplantation and the ultimate benefit to our patients.

Infertility is a prevalent global issue affecting approximately 17.5% of adults, with sole male factor contributing to 20–30% of cases. Oxidative stress (OS) is a critical factor in male infertility, disrupting the balance between reactive oxygen species (ROS) and antioxidants. This imbalance detrimentally affects sperm function and viability, ultimately impairing fertility. OS also triggers molecular changes in sperm, including DNA damage, lipid peroxidation, and alterations in protein expression, further compromising sperm functionality and potential fertilization. Diagnostic tools discussed in this review offer insights into OS markers, antioxidant levels, and intracellular ROS concentrations. By accurately assessing these parameters, clinicians can diagnose male infertility more effectively and thus tailor treatment plans to individual patients. Additionally, this review explores various treatment options for males with OS-associated infertility, such as empirical drugs, antioxidants, nanoantioxidants, and lifestyle modifications. By addressing the root causes of male infertility and implementing targeted interventions, clinicians can optimize treatment outcomes and enhance the chances of conception for couples struggling with infertility.

Abstract Fermionic artificial matter realized with cold atoms grants access to an unprecedented degree of control on sophisticated many-body effects with an enhanced flexibility of the operating conditions. Here, we consider three-component fermions with attractive interactions to study the formation of complex bound states, whose nature goes beyond the standard fermion pairing occurring in quantum materials. Such systems display clear analogies with quark matter. We address the nature of the bound states of a three-component fermionic system in a ring-shaped trap through the persistent current. In this way, we demonstrate that we can distinguish between color superfluid and trionic bound states. By analyzing finite temperature effects, we show how finite temperature can lead to the deconfinement of bound states. For weak interactions, the deconfinement occurs because of scattering states. In this regime, the deconfinement depends on the trade-off between interactions and thermal fluctuations. For strong interactions the features of the persistent current result from the properties of a suitable gas of bound states.

Abstract Background Combination anti‐obesity medications (AOMs) to treat postoperative bariatric surgery weight regain have limited data on their use in the clinical setting. Understanding the optimal treatment protocol in this cohort will maximize weight loss outcomes. Methods A retrospective review of bariatric surgery patients (N = 44) presenting with weight regain at a single academic multidisciplinary obesity center who were prescribed AOM(s) plus intensive lifestyle modification for 12 months. Results Age: 28–76 years old, 93% female, mean weight 110.2 ± 20.3 kg, BMI 39.7 ± 7.4 kg/m2, presenting 5.2 ± 1.6 years post‐bariatric surgery [27 (61.4%), 14 (31.8%), and 3 (6.8%) laparoscopic Roux‐en‐Y gastric bypass (RYGB), laparoscopic vertical sleeve gastrectomy (VSG), and open RYGB, respectively], with 15.1 ± 11.1 kg mean weight gain from nadir. Mean weight loss after medical intervention at 3‐, 6‐, and 12‐month time points was 4.4 ± 4.6 kg, 7.3 ± 7.0 kg, and 10.7 ± 9.2 kg, respectively. At 12 months, individuals prescribed 3 or more AOMs lost more weight than those prescribed one (−14.5 ± 9.0 kg vs. −4.9 ± 5.7 kg, p

Death investigation on tribal lands and of American Indian/Alaska Native (AIAN) people is complex and not well documented. An analysis of data from the 2018 Census of Medical Examiner and Coroner Offices (CMEC) provides a timely update on the extent of medicolegal death investigations (MDIs) on federal and state-recognized tribal lands. An estimated 150 MEC offices serve tribal lands, however, 44 % of these offices (i.e., 4 % of MEC offices) do not track cases from tribal lands separately. MEC offices with a population of 25,000 to 250,000 that serve tribal lands had more resources and access to information to perform MDIs than all other MEC offices. Analysis also indicates that the median number of unidentified human remains cases from MECs serving tribal lands is 6 times higher than that of jurisdictions not serving tribal lands. This analysis begins to elucidate gaps in the nation's understanding of MDI on tribal lands.

Abstract There are many examples of hybrid simulation models in healthcare education which are designed to simulate specific scenarios. However, there appears to be a need for a cost effective and configurable hybrid simulation platform which can be used by educators of various healthcare disciplines to simulate different scenarios. The purpose of this paper is to develop a proof-of-concept platform that can be easily implemented at little cost and provide flexibility to healthcare instructors to develop a variety of simulation scenarios, and to determine the effectiveness of this platform. Using a standardized patient, a person acting as a patient in a scripted manner, along with wearable and web-based technologies, a congestive heart failure simulation was used as an evaluative exercise for a group of personal support worker students at a Canadian Community College. Personal support workers typically provide care to any person who may require personal assistance with activities of daily living such as feeding, lifting, bathing, skin care and oral hygiene to name a few. Standardized patients are typically used in healthcare education to educate and evaluate soft skills, such as caregiver to patient communication, professionalism, as well as hard skills, such as history taking, examination and diagnostic skills (Rosen in J Crit Care 23:157–166, 2008). Instructor feedback indicated that the platform was easy to use and capable of simulating a large variety of scenarios. Pre and post test results are evidence of initial findings of promise indicating that the platform seemed to be effective in enabling students to meet learning outcomes. Focus group results seem to indicate an increase in student confidence as it relates to their ability to handle a similar scenario in the workplace.

Corticotropin-releasing factor (CRF) is essential for coordinating endocrine and neural responses to stress, frequently facilitated by vasopressin (AVP). Previous work has linked CRF hypersecretion, binding site changes, and dysfunctional serotonergic transmission with anxiety and affective disorders, including clinical depression. Crucially, CRF can alter serotonergic activity. In the dorsal raphé nucleus and serotonin (5-HT) terminal regions, CRF effects can be stimulatory or inhibitory, depending on the dose, site, and receptor type activated. Prior stress alters CRF neurotransmission and CRF-mediated behaviors. Lateral, medial, and ventral subdivisions of the central nucleus of the amygdala (CeA) produce CRF and coordinate stress responsiveness. The purpose of these experiments was to determine the effect of intracerebroventricular (icv) administration of CRF and AVP on extracellular 5-HT as an index of 5-HT release in the CeA, using in vivo microdialysis in freely moving rats and high performance liquid chromatography (HPLC) analysis. We also examined the effect of prior stress (1 h restraint, 24 h prior) on CRF- and AVP-mediated release of 5-HT within the CeA. Our results show that icv CRF infusion in unstressed animals had no effect on 5-HT release in the CeA. Conversely, in rats with prior stress, CRF caused a profound dose-dependent decrease in 5-HT release within the CeA. This effect was long-lasting (240 min) and was mimicked by CRF plus AVP infusion without stress. Thus, prior stress and AVP functionally alter CRF-mediated neurotransmission and sensitize CRF-induced inhibition of 5-HT release, suggesting that this is a potential mechanism underlying stress-induced affective reactivity in humans.

Background and Objectives: Obesity is a significant risk factor for hypogonadism and infertility that is further associated with reduced semen quality. The aim of this study is to evaluate the effect of clomiphene citrate (CC), prescribed for treating infertility, on serum testosterone and semen parameters, particularly in oligospermic obese hypogonadal men. Materials and Methods: A retrospective analysis of data related to men (n = 53) who underwent CC treatment for infertility and hypogonadism (testosterone < 300 ng/dL) was performed. Patients with obesity (BMI ≥ 30 kg/m2) and sperm concentration ≤ 15 × 106/mL were included for analysis. Results: The overall results showed that, in oligospermic obese men (n = 31), treatment with CC significantly improved baseline sperm concentration (4.5 ± 6.8 × 106/mL vs. 11.4 ± 15.5 × 106/mL, p < 0.05) and motility (31.5% ± 21.5% vs. 42.6% ± 14.7%, p < 0.05). Furthermore, subsequent examination of oligospermic hypogonadal obese men treated with CC (n = 13) revealed substantial improvements in baseline serum testosterone levels (193.8 ± 59.3 ng/dL vs. 332.7 ± 114.8 ng/dL, p < 0.05) along with an increase in sperm concentration, total motility, and normal morphology. Conclusions: The results of this retrospective study suggest that CC treatment not only improves chances of fertility outcomes by substantially improving semen parameters but also increases total serum testosterone levels in oligospermic obese men without any supplemental and expensive testosterone replacement therapy.

The location of corticotropin-releasing hormone receptor 2 (CRH-R2) on thyrotropes within the avian anterior pituitary (APit) and its activation by different stressors indicate a possible communication between hypothalamo-pituitary-adrenal (HPA) and thyroid (HPT) axes. Therefore, an experiment was designed to 1) compare the timing of major components of the HPT axis to those of the HPA axis; 2) address whether stressors activating the HPA axis may simultaneously upregulate components of the HPT axis. Blood, brain, and APit were sampled from chicks prior to stress (control) and 15, 30, 60, 90, and 120 min following immobilization (IM) stress. The nucleus of the hippocampal commissure (NHpC) and paraventricular nucleus (PVN) were cryo-dissected from brains for RT-qPCR. Gene expression of thyrotropin-releasing hormone (TRH) and its receptors (TRH-R1 and TRH-R3), urocortin3 (UCN3), deiodinase 2 (D2), and the second type of corticotropin-releasing hormone (CRH2) within the NHpC and PVN was measured. Additionally, gene expression of TRH receptors, thyroid stimulating hormone subunit beta (TSHβ), and D2 was determined in the APit and corticosterone assayed in blood. In brains, a significant upregulation in examined genes occurred at different times of IM. Specifically, UCN3 and CRH2 which have a high affinity to CRH-R2 showed a rapid increase in their mRNA levels that were accompanied by an early upregulation of TRHR1 in the NHpC. In the APit, a significant increase in gene expression of TSHβ and TRH receptors was observed. Therefore, results supported concurrent activation of major brain and APit genes associated with the HPA and HPT axes following IM. The initial neural gene expression originating within the NHpC resulted in the increase of TSHβ mRNA in the APit. Specifically, the rapid upregulation of UCN3 in the NHpC appeared responsible for the early activation of TSHβ in the APit. While sustaining TSHβ activation appeared to be due to both CRH2 and TRH. Therefore, data indicate that CRH-producing neurons and corticotropes as well as CRH- and TRH-producing neurons and thyrotropes are activated to produce the necessary energy required to maintain homeostasis in birds undergoing stress. Overall, data support the inclusion of the NHpC in the classical avian HPA axis and for the first time show the concurrent activation of the HPA axis and components of the HPT axis following a psychogenic stressor.

Porcine islets surviving the acute injury caused by humoral rejection and IBMIR will be subjected to cellular xenograft rejection, which is predominately mediated by CD4+ T cells and is characterised by significant infiltration of macrophages, B cells and T cells (CD4+ and CD8+). Overall, the response is different compared to the alloimmune response and more difficult to suppress. Activation of CD4+ T cells is both by direct and indirect antigen presentation. After activation they recruit macrophages and direct B cell responses. Although they are less important than CD4+ T cells in islet xenograft rejection, macrophages are believed to be a major effector cell in this response. Rodent studies have shown that xenoantigen-primed and CD4+ T cell-activated macrophages were capable of recognition and rejection of pancreatic islet xenografts, and they destroyed a graft via the secretion of various proinflammatory mediators, including TNF-α, reactive oxygen and nitrogen species, and complement factors. B cells are an important mediator of islet xenograft rejection via xenoantigen presentation, priming effector T cells and producing xenospecific antibodies. Depletion and/or inhibition of B cells combined with suppressing T cells has been suggested as a promising strategy for induction of xeno-donor-specific T- and B-cell tolerance in islet xenotransplantation. Thus, strategies that expand the influence of regulatory T cells and inhibit and/or reduce macrophage and B cell responses are required for use in combination with clinical applicable immunosuppressive agents to achieve effective suppression of the T cell-initiated xenograft response.

Xenotransplantation using porcine donors is rapidly approaching clinical applicability as an alternative therapy for treatment of many end-stage diseases including type 1 diabetes. Porcine neonatal islet cell clusters (NICC) have normalised blood sugar levels for relatively short periods in the preclinical diabetic rhesus model but have met with limited success in the stringent baboon model. Here we report that NICC from genetically modified (GM) pigs deleted for αGal and expressing the human complement regulators CD55 and CD59 can cure diabetes long-term in immunosuppressed baboons, with maximum graft survival exceeding 22 months. Five diabetic baboons were transplanted intraportally with 9,673 – 56,913 islet equivalents (IEQ) per kg recipient weight. Immunosuppression consisted of T cell depletion with an anti-CD2 mAb, tacrolimus for the first 4 months, and maintenance with belatacept and anti-CD154; no anti-inflammatory treatment or cytomegalovirus (CMV) prophylaxis/treatment was given. This protocol was well tolerated, with all recipients maintaining or gaining weight. Recipients became insulin-independent at a mean of 87 ± 43 days post-transplant and remained insulin-independent for 397 ± 174 days. Maximum graft survival was 675 days. Liver biopsies showed functional islets staining for all islet endocrine components, with no evidence of the inflammatory blood-mediated inflammatory reaction (IBMIR) and minimal leukocytic infiltration. The costimulation blockade-based immunosuppressive protocol prevented an anti-pig antibody response in all recipients. In conclusion, we demonstrate that genetic modification of the donor pig enables attenuation of early islet xenograft injury, and in conjunction with judicious immunosuppression provides excellent long-term function and graft survival in the diabetic baboon model.

Research into the potential associations between tinnitus and cognition has investigated specific cognitive domains in laboratory settings despite adults with tinnitus reporting broad cognitive difficulties in every-day life. To address this limitation, the present study compared performance and perceived exertion on a visual office-like task in 38 adults with tinnitus (19 with normal hearing and 19 with hearing loss) and 38 adults without tinnitus (19 with normal hearing and 19 with hearing loss) matched for age, sex and educational background. All participants were also assessed for hearing, anxiety and depression, and participants with tinnitus were also assessed for tinnitus handicap. No associations were found between presence of tinnitus and cognitive performance (mean total rate correct score on the visual office-like task being 2.9 for the tinnitus group, 2.8 for the control group, p = 0.612) and perceived exertion (mean ratings of perceived exertion on the Borg CR10-scale being 5.8 for the tinnitus group, 6.5 for the control group, p = 0.063) on the visual office-like task when corrected for standard (0.125 to 8 kHz) and extended high frequency (10 to 16 kHz) hearing thresholds, anxiety, and depression. The correction for extended high frequency average (10, 12.5, 14, and 16 kHz) hearing threshold was significant for performance (p = 0.009) but not perceived exertion on the visual office-like task. Overall, the results showed extended high frequency hearing, but not tinnitus, was associated with every-day cognitive performance. This indicates clinical testing of hearing thresholds above 8 kHz could support clinicians’ identification and management of cognitive difficulties. One management method suggested by the current findings would include provision of auditory stimulation at frequencies exceeding the frequency response of many current hearing aids.

Antibodies that target immune checkpoints such as cytotoxic T lymphocyte antigen 4 (CTLA‐4) and the programmed cell death protein 1/ligand 1 (PD-1/PD-L1) are now a treatment option for multiple cancer types. However, as a monotherapy, objective responses only occur in a minority of patients. Chemotherapy is widely used in combination with immune checkpoint blockade (ICB). Although a variety of isolated immunostimulatory effects have been reported for several classes of chemotherapeutics, it is unclear which chemotherapeutics provide the most benefit when combined with ICB. We investigated 10 chemotherapies from the main canonical classes dosed at the clinically relevant maximum tolerated dose in combination with anti‐CTLA-4/anti-PD-L1 ICB. We screened these chemo-immunotherapy combinations in two murine mesothelioma models from two different genetic backgrounds, and identified chemotherapies that produced additive, neutral or antagonistic effects when combined with ICB. Using flow cytometry and bulk RNAseq, we characterized the tumor immune milieu in additive chemo-immunotherapy combinations. 5-fluorouracil (5-FU) or cisplatin were additive when combined with ICB while vinorelbine and etoposide provided no additional benefit when combined with ICB. The combination of 5-FU with ICB augmented an inflammatory tumor microenvironment with markedly increased CD8+ T cell activation and upregulation of IFNγ, TNFα and IL-1β signaling. The effective anti‐tumor immune response of 5-FU chemo-immunotherapy was dependent on CD8+ T cells but was unaffected when TNFα or IL-1β cytokine signaling pathways were blocked. Our study identified additive and non-additive chemotherapy/ICB combinations and suggests a possible role for increased inflammation in the tumor microenvironment as a basis for effective combination therapy.

Machine learning (ML)-workflows enable unprejudiced/robust evaluation of complex datasets. Here, we analyzed over 490,000,000 data points to compare 10 different ML-workflows in a large (N=11,652) training dataset of human pancreatic single-cell (sc-)transcriptomes to identify genes associated with the presence or absence of insulin transcript(s). Prediction accuracy/sensitivity of each ML-workflow was tested in a separate validation dataset (N=2,913). Ensemble ML-workflows, in particular Random Forest ML-algorithm delivered high predictive power (AUC=0.83) and sensitivity (0.98), compared to other algorithms. The transcripts identified through these analyses also demonstrated significant correlation with insulin in bulk RNA-seq data from human islets. The top-10 features, (including IAPP, ADCYAP1, LDHA and SST) common to the three Ensemble ML-workflows were significantly dysregulated in scRNA-seq datasets from Ire-1αβ-/- mice that demonstrate dedifferentiation of pancreatic β-cells in a model of type 1 diabetes (T1D) and in pancreatic single cells from individuals with type 2 Diabetes (T2D). Our findings provide direct comparison of ML-workflows in big data analyses, identify key elements associated with insulin transcription and provide workflows for future analyses.

Abstract Background Available studies are lacking in analysis of baseline demographics and hospital presentation of patients at risk of expiring due to coronavirus disease 2019 (COVID-19), particularly Black American patients. We conducted a retrospective chart review to determine similarities in demographics and hospital presentation among patients who expired due to COVID-19 at an academic medical center in Brooklyn, New York. Study design and methods This is a retrospective observational study of 200 patients who expired due to complications of COVID-19. Patients were included in this study if they had laboratory-confirmed SARS-CoV-2 infection and expired during their admission. Data were collected on patients who expired between March 17 and April 16, 2020. Results A vast majority of patients were Black Americans (89%) with no history of international travel who had more than one comorbidity (81%), with the most common comorbidities being hypertension (84·5%), diabetes mellitus (57·5%), and obesity (41·5%). Fifty-five percent of our patient population had three or more comorbidities. Among patients with available data, C-reactive protein, lactate dehydrogenase, and ferritin values were elevated above normal limits at admission. Dyspnea was the most common presenting symptom (92·5%). Most (90·5%) presented within the first week of symptoms, with a median time of symptoms prior to expiration being 8·42 days (IQR 5·57–12·72). Interpretation Socioeconomic status and healthcare inequalities have greatly affected the Black population of Brooklyn, New York, and these disparities become even more apparent in COVID-19 infection. Patients presenting with numerous comorbidities and elevated inflammatory markers represent a population at high risk of in-hospital mortality.

Abstract Health-care education based upon technology enabled mannequins (high-fidelity simulators) is a costly investment for colleges and universities. However, a hybrid model using wearable technology integrated with human actors (standardized patients) may present a cost-effective alternative to high fidelity simulation training scenarios. A systematic literature review of papers published from 1960 to 2019 illustrates that hybrid simulation can be as effective as high fidelity simulators in certain training scenarios while at the same time providing a superior training context to enhance learners patient to care-giver interactions and to better immerse the trainee in the feelings and emotion of the scenario.

Maintaining a diverse urban forest that provides ecosystem services can promote urban sustainability and resilience to environmental change. Around the world, cities have taken to inventorying their urban trees and quantifying their ecosystem services but more so in industrialized counties than in Latin America. Here we describe the results of an i-Tree inventory that established 206 survey plots in the National Municipal District of Santo Domingo (NMDSD). We used social-ecological theory to evaluate potential factors that may influence urban forest structure, composition, and ecosystem services diversity across three wards with distinct social and urban characteristics. Rarefaction curves showed a diverse urban forest dominated by non-native trees that have ornamental and medicinal uses. Wards differed in species composition with palms being particularly dominant in Wards 1 and 2 where the proportion of low-income houses is smaller. Ward 1 supports high-income residential areas and Ward 3 is the area with higher population and housing densities and lower income residents. On average, we found no significant differences among wards in tree species richness, average dbh, leaf area, and percent tree cover per plot. Trees in Ward 2 were taller, on average, than those in Ward 1 but were comparable to those in Ward 3. Likewise, tree density per plot was highest in Ward 2, followed by Ward 1 and Ward 3. Despite these significant differences in stem densities, average values in four ecosystem services involving measures of carbon, rainfall, and contaminants (C-sequestration, C-storage, avoided runoff, and removal of air pollutants) were non-significant across wards. We found disproportionately more street trees in Ward 1 relative to Wards 2 and 3 and more trees in public spaces in Wards 1 and 2 relative to Ward 3. Evidence for the luxury effect on tree distribution in the NMDSD was subtle and manifested mostly through differences in species composition and tree distribution across public and private domains as well as the amount of planting space. Overall results point to inequalities in the potential of reforestation among NMDS wards and an overabundance of non-native species, which should guide urban forest management with ecosystem services and conservation goals.

Background & Aims: Pancreatic islet β-cells are factories for insulin production; however, ectopic expression of insulin also is well recognized. The gallbladder is a next-door neighbor to the developing pancreas. Here, we wanted to understand if gallbladders contain functional insulin-producing cells. Methods: We compared developing and adult mouse as well as human gallbladder epithelial cells and islets using immunohistochemistry, flow cytometry, enzyme-linked immunosorbent assays, RNA sequencing, real-time polymerase chain reaction, chromatin immunoprecipitation, and functional studies. Results: We show that the epithelial lining of developing, as well as adult, mouse and human gallbladders naturally contain interspersed cells that retain the capacity to actively transcribe, translate, package, and release insulin. We show that human gallbladders also contain functional insulin-secreting cells with the potential to naturally respond to glucose in vitro and in situ. Notably, in a non-obese diabetic (NOD) mouse model of type 1 diabetes, we observed that insulin-producing cells in the gallbladder are not targeted by autoimmune cells. Interestingly, in human gallbladders, insulin splice variants are absent, although insulin splice forms are observed in human islets. Conclusions: In summary, our biochemical, transcriptomic, and functional data in mouse and human gallbladder epithelial cells collectively show the evolutionary and developmental similarities between gallbladder and the pancreas that allow gallbladder epithelial cells to continue insulin production in adult life. Understanding the mechanisms regulating insulin transcription and translation in gallbladder epithelial cells would help guide future studies in type 1 diabetes therapy.

Antecedentes: El municipio guatemalteco de Nebaj es una comunidad indígena maya con tasas de suicidio percibidas como superiores al promedio nacional. Objetivos: Este artículo evalúa las percepciones de los factores relacionados con el suicidio en Nebaj para informar las estrategias de prevención frente a este. Métodos: Esta investigación utiliza métodos cualitativos y cuantitativos. Los datos provienen de 19 entrevistas semiestructuradas con partes interesadas en Nebaj, junto con revisiones de casos de registros administrativos oficiales de la Policía Nacional Civil (pnc) y estadísticas agregadas de registros oficiales de suicidios recopilados por la pnc y el hospital de Nebaj. Resultados: Las partes interesadas locales vinculan el suicidio con deudas pendientes, la disolución de relaciones románticas, problemas de salud mental, conflictos intrafamiliares y abuso de sustancias. A nivel comunitario, los entrevistados relacionan la disolución de familias a través de la migración y las deudas resultantes. Limitaciones: Las instituciones públicas que abordan el suicidio están aisladas y no trabajan juntas ni recopilan datos con los mismos fines. Los datos disponibles están fragmentados y son difíciles de contrastar entre fuentes. Conclusión: Las entrevistas revelan temas comunes en la percepción de factores contribuyentes al suicidio en Nebaj. Los datos empíricos muestran que es más probable que los jóvenes participen en intentos de suicidio fatales y no fatales. Los análisis también indican que el método de suicidio más común es la ingestión de pesticidas agrícolas.

Background Children with Fontan circulation are known to be at increased risk for neurodevelopmental problems and decreased health‐related quality of life (HRQOL), but many factors that may contribute to this risk are unknown. Sleep disturbances may be one previously unidentified factor that contributes to this risk. Methods and Results We analyzed data from the Pediatric Heart Network Fontan cross‐sectional study to evaluate associations between a parent or child report of sleep disturbance with reported neurodevelopmental concerns and HRQOL in 558 children with Fontan circulation. Parent‐reported sleep disturbance was present in 11% of participants and child‐reported sleep disturbance was present in 15%. Parent‐reported sleep disturbance was associated with a significantly higher risk of attention problems, anxiety, depression, behavioral problems, and developmental delay (P

(1) Background: Systemic antibiotic use in chronic wounds is alarmingly high worldwide. Between 53% to 71% of patients are prescribed at least one course per chronic wound. Systemic antibiotic use should follow antibiotic stewardship guidelines and ought to be reserved for situations where their use is deemed supported by clinical indications. Unfortunately, in the field of wound care, indiscriminate and often inadequate use of systemic antibiotics is leading to both patient complications and worsening antibiotic resistance rates. Implementing novel tools that help clinicians prevent misuse or objectively determine the true need for systemic antibiotics is essential to reduce prescribing rates. (2) Methods: We present a compendium of available systemic antibiotic prescription rates in chronic wounds. The impact of various strategies used to improve these rates, as well as preliminary data on the impact of implementing fluorescence imaging technology to finesse wound status diagnosis, are presented. (3) Results: Interventions including feedback from wound care surveillance and treatment data registries as well as better diagnostic strategies can ameliorate antibiotic misuse. (4) Conclusions: Interventions that mitigate unnecessary antibiotic use are needed. Effective strategies include those that raise awareness of antibiotic overprescribing and those that enhance diagnosis of infection, such as fluorescence imaging.

Seminal oxidative stress and sperm DNA damage are potential etiologies of male factor infertility. The present study aims to evaluate the relationship between oxidation-reduction potential (ORP), a measure of oxidative stress, and sperm DNA fragmentation (SDF) by conducting a systematic review and meta-analysis of relevant clinical data. A literature search was performed according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. The COVIDENCE tool was used to screen and identify studies evaluating seminal ORP and SDF. Studies (n = 7) that measured seminal ORP and SDF of 3491 semen samples were included in the analysis. The fixed-effects model revealed a significant pooled correlation coefficient (r = 0.24; p < 0.001) between seminal ORP and SDF. Furthermore, subgroup analyses indicated that the pooled correlation coefficient between ORP and sperm chromatin dispersion (SCD) assay was less than other SDF assays (0.23 vs. 0.29). There was a moderate level of heterogeneity (I2 = 42.27%) among the studies, indicating a lack of publication bias. This is the first meta-analysis to reveal a positive correlation between seminal ORP and SDF. Furthermore, this study indicates the role of oxidative stress in the development of sperm DNA damage and thus warrants prospectively exploring the clinical value of these sperm function tests.

Social rank functions to facilitate coping responses to socially stressful situations and conditions. The evolution of social status appears to be inseparably connected to the evolution of stress. Stress, aggression, reward, and decision-making neurocircuitries overlap and interact to produce status-linked relationships, which are common among both male and female populations. Behavioral consequences stemming from social status and rank relationships are molded by aggressive interactions, which are inherently stressful. It seems likely that the balance of regulatory elements in pro- and anti-stress neurocircuitries results in rapid but brief stress responses that are advantageous to social dominance. These systems further produce, in coordination with reward and aggression circuitries, rapid adaptive responding during opportunities that arise to acquire food, mates, perch sites, territorial space, shelter and other resources. Rapid acquisition of resources and aggressive postures produces dominant individuals, who temporarily have distinct fitness advantages. For these reasons also, change in social status can occur rapidly. Social subordination results in slower and more chronic neural and endocrine reactions, a suite of unique defensive behaviors, and an increased propensity for anxious and depressive behavior and affect. These two behavioral phenotypes are but distinct ends of a spectrum, however, they may give us insights into the troubling mechanisms underlying the myriad of stress-related disorders to which they appear to be evolutionarily linked.

The Fontan procedure was created to address the mixing of pulmonary and systemic venous return in patients with a single functional ventricle. The patient in this case with a Fontan repair experienced multiple pulmonary emboli 10 days post-partum. We outline management and recommendations when treating these patients. (Level of Difficulty: Beginner.)

CARD9 and CARD11 propagate signaling by nucleating Bcl10 polymerization in immune cells and are both held in an autoinhibited state prior to activation. Here, the authors combine structural, biochemical, and cell-based approaches to reveal the structural basis for CARD9/11 autoinhibition and show that the two proteins are activated through similar but distinct mechanisms.

Abstract Background Telemetry and biologging systems, ‘tracking’ hereafter, have been instrumental in meeting the challenges associated with studying the ecology and behaviour of cryptic, wide-ranging marine mega-vertebrates. Over recent decades, globally, sea turtle tracking has increased exponentially, across species and life-stages, despite a paucity of studies investigating the effects of such devices on study animals. Indeed, such studies are key to informing whether data collected are unbiased and, whether derived estimates can be considered typical of the population at large. Methods Here, using a 26-year individual-based monitoring dataset on sympatric green (Chelonia mydas) and loggerhead (Caretta caretta) turtles, we provide the first analysis of the effects of device attachment on reproduction, growth and survival of nesting females. Results We found no significant difference in growth and reproductive correlates between tracked and non-tracked females in the years following device attachment. Similarly, when comparing pre- and post-tracking data, we found no significant difference in the reproductive correlates of tracked females for either species or significant carry-over effects of device attachment on reproductive correlates in green turtles. The latter was not investigated for loggerhead turtles due to small sample size. Finally, we found no significant effects of device attachment on return rates or survival of tracked females for either species. Conclusion While there were no significant detrimental effects of device attachment on adult sea turtles in this region, our study highlights the need for other similar studies elsewhere and the value of long-term individual-based monitoring.

Summary: Dicer knockout mouse models demonstrated a key role for microRNAs in pancreatic β-cell function. Studies to identify specific microRNA(s) associated with human (pro-)endocrine gene expression are needed. We profiled microRNAs and key pancreatic genes in 353 human tissue samples. Machine learning workflows identified microRNAs associated with (pro-)insulin transcripts in a discovery set of islets (n = 30) and insulin-negative tissues (n = 62). This microRNA signature was validated in remaining 261 tissues that include nine islet samples from individuals with type 2 diabetes. Top eight microRNAs (miR-183-5p, -375-3p, 216b-5p, 183-3p, -7-5p, -217-5p, -7-2-3p, and -429-3p) were confirmed to be associated with and predictive of (pro-)insulin transcript levels. Use of doxycycline-inducible microRNA-overexpressing human pancreatic duct cell lines confirmed the regulatory roles of these microRNAs in (pro-)endocrine gene expression. Knockdown of these microRNAs in human islet cells reduced (pro-)insulin transcript abundance. Our data provide specific microRNAs to further study microRNA-mRNA interactions in regulating insulin transcription.

Erectile dysfunction (ED) is both a common and complex disease process. Existing ED treatments do not always achieve adequate results. There is clinical interest in employing regenerative therapies, including low-intensity extracorporeal shockwave therapy (Li-ESWT), platelet rich plasma (PRP), and stem cell therapy (SCT), in the treatment of ED as adjunct or alternative treatments. Here, we present evidence for emerging shockwave- and cell-based regenerative therapies for the treatment of ED following a thorough review of the existing PubMed literature pertaining to Li-ESWT, PRP, and SCT in relation to the treatment of ED. Li-ESWT causes microtrauma in tissue that hypothetically upregulates angiogenesis and recruits stem cells. Several large-scale systematic reviews and meta-analyses have reported that Li-ESWT improved ED in humans. Additionally, evidence has commenced to show that Li-ESWT may be effective against two recognized and complex etiologies of ED: diabetic and neurogenic. PRP delivers an autologous sample rich in growth factors to damaged tissue. Animal model studies have demonstrated improved erectile function recovery as well as preservation of cavernous nerve axons. Studies with PRP in humans are limited. SCT utilizes the regenerative potential of stem cells for healing of damaged tissue. In the treatment of ED, SCT has been used in the setting of diabetic and post-prostatectomy ED. Results of human studies are varied, although SCT treatments did result in increased erectile rigidity with some patients recovering the ability to achieve penetration. While these regenerative therapies show potential to augment the current treatment regimen for ED, there is a paucity of evidence to support the safety and efficacy of these treatments. Further research is necessary to define the role of these alternative therapies in the treatment of ED.

Background: This study continued the development of an isiZulu speech reception threshold (zSRT) test for use with first language, adult speakers of isiZulu. Objectives: The objective of this study was to determine the convergent and concurrent validity of the zSRT test. Methods: One hundred adult isiZulu first-language speakers with normal hearing and 76 first-language, adult isiZulu speakers with conductive or sensorineural hearing losses ranging from mild to severe were assessed on pure tone audiometry and a newly developed isiZulu SRT test. Convergent validity was established through agreement of the zSRT scores with pure tone average (PTA) scores. Concurrent validity was assessed by examining the steepness of the psychometric curve for each word in the zSRT test for each type and degree of hearing loss. Results: Intraclass correlation coefficient analyses showed zSRT scores were in substantial to very high agreement with PTA scores for the normal hearing and hearing loss groups (NH – right ear ICC consistency = 0.78, left ear ICC = 0.67; HL – right ear ICC consistency = 0.97, left ear ICC consistency = 0.95). The mean psychometric slope (%/dB) at 50% correct perception for all words in the zSRT test was 4.92%/dB for the mild conductive hearing loss group, 5.26%/dB for the moderate conductive hearing loss group, 2.85%/dB for the moderately severe sensorineural hearing loss group and 2.47%/dB for the severe sensorineural hearing loss group. These slopes were appropriate for the degree of hearing loss observed in each group. Conclusion: The zSRT test showed convergent and concurrent validity for assessing SRT in first language, adult speakers of isiZulu.

Simultaneous pancreas-kidney (SPK) transplantation remains the most effective treatment for providing consistent and long-term euglycemia in patients having type 1 diabetes with renal failure. Thrombosis of the pancreatic vasculature continues to contribute significantly to early graft failure and loss. We compared the rate of thrombosis to graft loss and systematically reviewed risk factors impacting early thrombosis of the pancreas allograft following SPK transplantation. We searched the MEDLINE, EMBASE, The Cochrane Library, and PREMEDLINE databases for studies reporting thrombosis following pancreas transplantation. Identified publications were screened for inclusion and synthesized into a data extraction sheet. Sixty-three studies satisfied eligibility criteria: 39 cohort studies, 22 conference abstracts, and 2 meta-analyses. Newcastle-Ottawa Scale appraisal of included studies demonstrated cohort studies of low bias risk; 1127 thrombi were identified in 15 936 deceased donor, whole pancreas transplants, conferring a 7.07% overall thrombosis rate. Thrombosis resulted in pancreatic allograft loss in 83.3% of reported cases. This review has established significant associations between donor and recipient characteristics, procurement and preservation methodology, transplantation technique, postoperative management, and increased risk of early thrombosis in the pancreas allograft. Further studies examining the type of organ preservation fluid, prophylactic heparin protocol, and exocrine drainage method and early thrombosis should also be performed.

Background: The unique immunomodulatory capacity of helminth parasites has been investigated as a novel strategy in the prevention of allograft rejection after transplantation. This review was conducted to fully evaluate the specific effects of helminth therapy on allograft survival reported in published studies of animal models of allogeneic transplantation.Method: Following PRISMA protocol guidelines, a literature search was conducted using PubMed, MEDLINE via OvidSP, along with additional manual searches of selected reference lists. Publications describing helminth intervention within allograft transplantation models were screened for relevance to eligibility criteria. Primary and secondary outcomes were extracted using standardized data collection tables. The SYRCLE risk of bias assessment tool was used for quality assessment. Due to heterogeneity of study designs, meta-analysis could not be performed; rather outcomes are presented as a narrative synthesis with concept mapping. This review was registered in PROSPERO with ID: CRD42018097175.Results: The literature search generated 1,443 publications, which after screening for relevance to the eligibility criteria yielded 15 publications for qualitative analysis. All 15 publications reported improvement to allograft survival as a result of helminth therapy. This prolonged allograft survival was not significantly different when helminth-derived products were used compared to live infection. However, the extent of positive impact on allograft survival was noted to be dependent on study design factors, such as the chronicity of the live helminth infection, allograft type and the species/genus of helminth selected.Conclusion: Both live and product-based helminth therapy have potential applications as novel immune regulators or adjuncts for the prevention of allograft rejection. However, there were differences in efficacy between different worms and preparations of worm-derived products. Therefore, further studies are required to determine the most appropriate worm for a specific allograft, to elucidate the optimal dose and route of administration, and to better understand the modulation of immune responses that can mediate tolerance.

Introduction A number of studies have reported an association between peripheral hearing impairment, central auditory processing and Alzheimer’s disease (AD) and its preclinical stages. Both peripheral hearing impairment and central auditory processing disorders are observed many years prior to the clinical manifestation of AD symptoms, hence, providing a long window of opportunity to investigate potential interventions against neurodegenerative processes. This paper outlines the protocol for a systematic review of studies examining the central auditory processing functions in AD and its preclinical stages, investigated through behavioural (clinical assessments that require active participation) central auditory processing tests.Methods and analysis We will use the keywords and Medical Subject Heading terms to search the following electronic databases: MEDLINE, PsychINFO, PubMed, Scopus, EMBASE and CINAHL Plus. Studies including assessments of central auditory function in adults diagnosed with dementia, AD and its preclinical stages that were published before 8 May 2019 will be reviewed. This review protocol will be reported according to the Preferred Reporting Items for Systematic Reviews and Meta-Analyses Protocols guidelines. Data analysis and search results will be reported in the full review. This manuscript has designed the protocols for a systematic review that will identify the behavioural clinical central auditory processing measures that are sensitive to the changes in auditory function in adults with AD and its preclinical stages. Such assessments may subsequently help to design studies to examine the potential impact of hearing and communication rehabilitation of individuals at risk of AD.Ethics and dissemination Ethical approval is not required as this manuscript only reports the protocols for conducting a systematic review as primary data will only be reviewed and not be collected. The results of this systematic review will be disseminated through publication and in scientific conferences.PROSPERO registration number CRD42017078272.

Erich Meyerhoff was an academic health sciences librarian and a distinguished member of the Medical Library Association when he was invited to present the Janet Doe Lecture in 1977. His lecture on the state of the association is considered one of the finest Doe lectures and is still relevant more than forty years later, not only from an historical perspective, but also for his projections for the future and his prescient comments about the future of hospital librarianship and the important role of women in the association. Key 1977 Doe lecture topics are reviewed and updated in the context of the current health sciences library environment.

We study the persistent current in a system of SU($N$) fermions with repulsive interaction confined in a ring-shaped potential and pierced by an effective magnetic flux. By applying a combination of Bethe ansatz and numerical analysis, we demonstrate that, as a combined effect of spin correlations, interactions and applied flux a specific phenomenon can occur in the system: spinon creation in the ground state. As a consequence, peculiar features in the persistent current arise. The elementary flux quantum, which fixes the persistent current periodicity, is observed to evolve from a single particle one to an extreme case of fractional flux quantum, in which one quantum is shared by all the particles. We show that the persistent current depends on the number of spin components $N$, number of particles and interaction in a specific way that in certain physical regimes has universality traits. At integer filling fractions, the persistent current is suppressed above a threshold of the repulsive interaction by the Mott spectral gap. Despite its mesoscopic nature, the current displays a clear finite size scaling behavior. Specific parity effects in the persistent current landscape hold.

Dexamethasone is a synthetic glucocorticoid commonly used for the prevention and management of side effects in cancer patients undergoing chemotherapy. While it is effective as an anti-emetic and in preventing hypersensitivity reactions, dexamethasone depletes peripheral blood lymphocytes and impacts immune responses. The effect of dexamethasone on the number and quality of tumour-infiltrating leukocytes has not been reported. To address this, we calibrated the dose in two different strains of mice to achieve the same extent of peripheral blood lymphocyte depletion observed in patients with cancer. Doses that caused analogous depletion of T and B lymphocytes and NK cells from the peripheral blood, elicited no change in these populations within the tumour. The expression of immune checkpoint molecules PD-1, OX40, GITR and TIM3 on tumour-infiltrating lymphocytes was not altered. We found that dexamethasone had a small but significant deleterious impact on weakly efficacious chemoimmunotherapy but had no effect when the protocol was highly efficacious. Based on these results, we predict that dexamethasone will have a modest negative influence on the overall effectiveness of chemoimmunotherapy treatment.

Background. Normothermic machine perfusion (NMP) may allow resuscitation and improved assessment of kidneys before transplantation. Using discarded human kidneys, we investigated the mechanistic basis and translational potential of NMP compared with cold static storage (CS). Methods. Discarded deceased donor kidneys (n = 15) underwent 1-hour NMP following CS. Renal perfusion, biochemical, and histologic parameters were recorded. NMP was directly compared with CS in paired donor kidneys using simulated transplantation with allogeneic whole blood, followed by assessment of the aforementioned parameters, in addition to RNA sequencing. Results. Kidneys were successfully perfused, with improved renal blood flows and resistance over the course of perfusion, and evidence of urine output (median 21 mL), in all but one kidney. NMP completely resolved nonperfused regions in discarded donation after circulatory death kidneys. In paired kidneys (n = 4 pairs), transcriptomic analyses showed induction of stress and inflammatory pathways in NMP kidneys, with upregulation of pathways promoting cell survival and proliferation. Furthermore, the NMP pairs had significantly better renal perfusion (1.5–2 fold improvement) and functional parameters, and amelioration of cell death, oxidative stress, and complement activation. Conclusions. In this pilot preclinical study using simulated transplantation of paired kidneys, NMP of discarded marginal kidneys demonstrated some significant mechanistic benefits in comparison to CS alone. NMP may have potential to reduce organ discards and enhance early graft function in such kidneys.

An in vivo validation study was performed to confirm the accuracy of extraoral photobiomodulation therapy (PBMT) dosimetry determined by modelling. The Monte Carlo technique was utilized to calculate the fluence rate and absorbed power of light delivered through multi-layered tissue. Optical properties used during Monte Carlo simulations were taken from the literature. Morphological data of four study volunteers were acquired using magnetic resonance imaging (MRI) scans. Light emitting diode (LED) coupled to a power meter were utilized to measure transmitted power through each volunteer’s cheek, in vivo. The transmitted power determined by Monte Carlo modelling was compared to the in vivo measurements to determine the accuracy of the simulations. Experimental and simulation results were in good agreement for all four subjects. The difference between the mean values of the measured transmission was within 12% from the respective transmission obtained using Monte Carlo simulations. The results of the study indicate that Monte Carlo modelling is a robust and reliable method for light dosimetry.

Cold ischemia and hence travel time can adversely affect outcomes of islet isolation. The aim of this study was to compare the isolation and transplant outcomes of donor pancreata according to the distance from islet isolation facility. Principally, those within a 50 km radius of the centre were compared with those from regional areas within the state and those from interstate donors within Australia. Organ donors were categorised according to distance from National Pancreas Transplant Unit Westmead (NPTU). Donor characteristics were analysed statistically against islet isolation outcomes. These were age, BMI, cause and mechanism of death, days in ICU, gender, inotrope and steroid use, cold ischemia time (CIT) and retrieval surgical team. Between March 2007 and December 2020, 297 islet isolations were performed at our centre. A total of 149 donor pancreata were local area, and 148 non-local regions. Mean distance from the isolation facility was 780.05 km. Mean pancreas CIT was 401.07 ± 137.71 min and was significantly different between local and non-local groups (297.2 vs. 487.5 min, p < 0.01). Mean age of donors was 45.22 years, mean BMI was 28.82, sex ratio was 48:52 F:M and mean time in ICU was 3.07 days. There was no significant difference between local and non-local for these characteristics. The mean CIT resulting in islet transplantation was 297.1 ± 91.5 min and longest CIT resulting in transplantation was 676 min. There was no significant difference in islet isolation outcomes between local and non-local donors for characteristics other than CIT. There was also no significant effect of distance from the isolation facility on positive islet transplant outcomes (C-peptide > 0.2 at 1 month post-transplant). Conclusions: Distance from the isolation centre did not impact on isolation or transplant outcomes supporting the ongoing nationwide use of shipping pancreata for islet isolation and transplantation.

Variants in the small surface gene of hepatitis B virus (HBV), which codes for viral surface antigen (HBsAg), can affect the efficacy of HBsAg screening assays and can be associated with occult HBV infection (OBI). This study aimed to characterise the molecular diversity of the HBV small surface gene from HBV-reactive Australian blood donors. HBV isolates from 16 HBsAg-positive Australian blood donors’ plasma were sequenced and genotyped by phylogenies of viral coding genes and/or whole genomes. An analysis of the genetic diversity of eight HBV small surface genes from our 16 samples was conducted and compared with HBV sequences from NCBI of 164 international (non-Australian) blood donors. Genotypes A–D were identified in our samples. The region of HBV small surface gene that contained the sequence encoding the ‘a’ determinant had a greater genetic diversity than the remaining part of the gene. No escape mutants or OBI-related variants were observed in our samples. Variant call analysis revealed two samples with a nucleotide deletion leading to truncation of polymerase and/or large/middle surface amino acid sequences. Overall, we found that HBV small surface gene sequences from Australian donors demonstrated a lower level of genetic diversity than those from non-Australian donor population included in the study.

Abstract Background Left-sided lesions (LSLs) account for an important fraction of severe congenital cardiovascular malformations (CVMs). The genetic contributions to LSLs are complex, and the mutations that cause these malformations span several diverse biological signaling pathways: TGFB, NOTCH, SHH, and more. Here, we use whole exome sequence data generated in 342 LSL cases to identify likely damaging variants in putative candidate CVM genes. Methods Using a series of bioinformatics filters, we focused on genes harboring population-rare, putative loss-of-function (LOF), and predicted damaging variants in 1760 CVM candidate genes constructed a priori from the literature and model organism databases. Gene variants that were not observed in a comparably sequenced control dataset of 5492 samples without severe CVM were then subjected to targeted validation in cases and parents. Whole exome sequencing data from 4593 individuals referred for clinical sequencing were used to bolster evidence for the role of candidate genes in CVMs and LSLs. Results Our analyses revealed 28 candidate variants in 27 genes, including 17 genes not previously associated with a human CVM disorder, and revealed diverse patterns of inheritance among LOF carriers, including 9 confirmed de novo variants in both novel and newly described human CVM candidate genes (ACVR1, JARID2, NR2F2, PLRG1, SMURF1) as well as established syndromic CVM genes (KMT2D, NF1, TBX20, ZEB2). We also identified two genes (DNAH5, OFD1) with evidence of recessive and hemizygous inheritance patterns, respectively. Within our clinical cohort, we also observed heterozygous LOF variants in JARID2 and SMAD1 in individuals with cardiac phenotypes, and collectively, carriers of LOF variants in our candidate genes had a four times higher odds of having CVM (odds ratio = 4.0, 95% confidence interval 2.5–6.5). Conclusions Our analytical strategy highlights the utility of bioinformatic resources, including human disease records and model organism phenotyping, in novel gene discovery for rare human disease. The results underscore the extensive genetic heterogeneity underlying non-syndromic LSLs, and posit potential novel candidate genes and complex modes of inheritance in this important group of birth defects.

Abstract Background Cytotoxic chemotherapeutics form the cornerstone of systemic treatment of many cancers. Patients are dosed at maximum tolerated dose (MTD), which is carefully determined in phase I studies. In contrast, in murine studies, dosages are often based on customary practice or small pilot studies, which often are not well documented. Consequently, research groups need to replicate experiments, resulting in an excess use of animals and highly variable dosages across the literature. In addition, while patients often receive supportive treatments in order to allow dose escalation, mice do not. These issues could affect experimental results and hence clinical translation. Methods To address this, we determined the single-dose MTD in BALB/c and C57BL/6 mice for a range of chemotherapeutics covering the canonical classes, with clinical score and weight as endpoints. Results We found that there was some variation in MTDs between strains and the tolerability of repeated cycles of chemotherapy at MTD was drug-dependent. We also demonstrate that dexamethasone reduces chemotherapy-induced weight loss in mice. Conclusion These data form a resource for future studies using chemotherapy in mice, increasing comparability between studies, reducing the number of mice needed for dose optimisation experiments and potentially improving translation to the clinic.