Artificial chemotaxis under electrodiffusiophoresis.

HYPOTHESIS: Through a large parameter space, electric fields can tune colloidal interactions and forces leading to diverse static and dynamical structures. So far, however, field-driven interactions have been limited to dipole-dipole and hydrodynamic contributions. Nonetheless, in this work, we propose that under the right conditions, electric fields can also induce interactions based on local chemical fields and diffusiophoretic flows. EXPERIMENTS: Herein, we present a strategy to generate and measure 3D chemical gradients under electric fields. In this approach, faradaic reactions at electrodes induce global pH gradients that drive long-range transport through electrodiffusiophoresis. Simultaneously, the electric field induces local pH gradients by driving the particle's double layer far from equilibrium. FINDINGS: As a result, while global pH gradients lead to 2D focusing away from electrodes, local pH gradients induce aggregation in the third dimension. Evidence points to a mechanism of interaction based on diffusiophoresis. Interparticle interactions display a strong dependence on surface chemistry, zeta potential and diameter of particles. Furthermore, pH gradients can be readily tuned by adjusting the voltage and frequency of the electric field. For large Péclet numbers, we observed a collective chemotactic-like collapse of particles. Remarkably, such collapse occurs without reactions at a particle's surface. By mixing particles with different sizes, we also demonstrate, through experiments and Brownian dynamics simulations, the emergence of non-reciprocal interactions, where small particles are more drawn towards large ones.

A lytic transglycosylase connects bacterial focal adhesion complexes to the peptidoglycan cell wall.

The Gram-negative bacterium Myxococcus xanthus glides on solid surfaces. Dynamic bacterial focal adhesion complexes (bFACs) convert proton motive force from the inner membrane into mechanical propulsion on the cell surface. It is unclear how the mechanical force transmits across the rigid peptidoglycan (PG) cell wall. Here, we show that AgmT, a highly abundant lytic PG transglycosylase homologous to Escherichia coli MltG, couples bFACs to PG. Coprecipitation assay and single-particle microscopy reveal that the gliding motors fail to connect to PG and thus are unable to assemble into bFACs in the absence of an active AgmT. Heterologous expression of E. coli MltG restores the connection between PG and bFACs and thus rescues gliding motility in the M. xanthus cells that lack AgmT. Our results indicate that bFACs anchor to AgmT-modified PG to transmit mechanical force across the PG cell wall.

IL-7-dependent and -independent lineages of IL-7R-dependent human T cells.

Infants with biallelic IL7R loss-of-function variants have severe combined immune deficiency (SCID) characterized by the absence of autologous T lymphocytes, but normal counts of circulating B and NK cells (T-B+NK+ SCID). We report 6 adults (aged 22 to 59 years) from 4 kindreds and 3 ancestries (Colombian, Israeli Arab, Japanese) carrying homozygous IL7 loss-of-function variants resulting in combined immunodeficiency (CID). Deep immunophenotyping revealed relatively normal counts and/or proportions of myeloid, B, NK, and innate lymphoid cells. By contrast, the patients had profound T cell lymphopenia, with low proportions of innate-like adaptive mucosal-associated invariant T and invariant NK T cells. They also had low blood counts of T cell receptor (TCR) excision circles, recent thymic emigrant T cells and naive CD4+ T cells, and low overall TCR repertoire diversity, collectively indicating impaired thymic output. The proportions of effector memory CD4+ and CD8+ T cells were high, indicating IL-7-independent homeostatic T cell proliferation in the periphery. Intriguingly, the proportions of other T cell subsets, including TCRγδ+ T cells and some TCRαß+ T cell subsets (including Th1, Tfh, and Treg) were little affected. Peripheral CD4+ T cells displayed poor proliferation, but normal cytokine production upon stimulation with mitogens in vitro. Thus, inherited IL-7 deficiency impairs T cell development less severely and in a more subset-specific manner than IL-7R deficiency. These findings suggest that another IL-7R-binding cytokine, possibly thymic stromal lymphopoietin, governs an IL-7-independent pathway of human T cell development.

Intake and digestibility of four rations with different fiber levels in alpacas (Vicugna pacos).

The aim of this study was to evaluated the impact of different fibre levels in alpaca diet on voluntary feed intake and apparent digestibility, and to estimate the digestibility of organic matter (OMD) from the content of crude protein (CP) in feces. The study was carried out with twelve alpacas (36.7 ± 6.4 kg body weight- BW), which were offered 4 treatments with different neutral detergent fiber content (NDF. T1: 40.3%; T2: 62%; T3: 68%; T4: 72%) under a switch back design. Absolute daily dry matter intake (DMI) was higher for T1 (678 g/d) than T4 (312 g/d) (p ≥ 005). NDF intake was similar between treatments when related to BW or MW (on average 1% BW and 22 g/kg MW. p ≥ 0.05). Water intake (L/kg DMI) was higher in T1 compared to the other treatments, with values ranging from 2.9 L/kg DMI(T1) to 2.8 L/kg DMI(T4), respectively (p ≤ 0.05). Digestibility of dry matter, organic matter and CP was higher in T1 than in the other treatments, with average values ranging from 72% for T1 to 32% for T4 (p ≤ 0.05). NDF digestibility was similar among treatments (p ≥ 0.05). The regression equation generated to predict OMD (y) was as follows: y = 0.360 + 0.08294*fecal CP (g/kg OM). Further studies will indicate whether faecal nitrogen can be used to estimate digestibility and hence diet quality in South American camelids.

A Patient With Symptomatic Pheochromocytoma and Very Severe Aortic Valve Stenosis: Diagnostic and Management Approach.

Preoperative management of a pheochromocytoma involves α-adrenergic receptor blockers and volume expansion followed by ß-adrenergic receptor blockers and then adrenalectomy, which in the presence of very severe aortic valve stenosis would be challenging. We describe a patient with this rare combination who underwent pharmacotherapy followed by transcatheter aortic valve replacement and then adrenalectomy, culminating in a successful outcome for this patient.

Key Clinical and Histopathological Features of a Pterygium-Like Induced Lesion in a Rabbit Model.

Purpose: Surgery is the definitive treatment for pterygium; therefore, reliable animal models are required for translational research. The goal of this investigation was to establish a standardized preclinical model of pterygium-like lesion. Methods: A subconjunctival injection of fibroblasts (NIH3T3) and extracellular matrix was administered to 22 New Zealand rabbits. Clinical evaluation was assessed at different points, the severity of the lesion was scored according to four grades and correlated with the area of hyperemia and the histopathological findings on day 23. Results: Thirteen of 22 eyes (60%) developed pterygium-like lesions after 7 days and progressed through different grades. Initially, grade 3, characterized by an elevated and fleshiness conjunctiva with tortuous hyperemia, was evident on day 7. By day 15, lesion decreased to grade 2, with less elevation and hyperemia. Subsequent improvement was noted, with grade 1 on day 18. Finally, day 23 was marked by a white‒yellow lesion, classified as grade 4. The area of hyperemia increased from grade 2 to grade 3 (P < 0.05) and decreased from grade 3 to grade 4 (P ≤ 0.05). Histopathological analysis revealed a tendency toward increasing inflammation at grades 2, 3, and 4. There was a correlation between clinical features and the degree of inflammation. Conclusions: Subconjunctival injection of NIH3T3 and extracellular matrix induces a pterygium-like lesion that progresses across four grades, beginning with an acute inflammatory process that evolve a chronic form. This study provides a replicable model for simulating pterygium. Translational Relevance: The development of a standardized preclinical model of pterygium to evaluate new pharmacological or surgical treatments.

Closure of small fenestrations without optimal rims with a PDA coil far from primary atrial septal defects: a new approach.

An ostium secundum atrial septal defect (ASD) is a CHD that can be treated percutaneously since 1974, mostly cases with only one main defect. In cases with fenestrations close to the main defect, a single occluder can be used for treatment because the discs extend beyond the waist of the device. In some cases where the defects are far from each other, they may require either more than one device or surgical closure. We present two patients in whom we observed fenestrations far from the primary defect. Initially, the main ASDs were closed with an ASD occluder, and then the fenestrations were closed with a patent ductus arteriosus (PDA) coil, resulting in complete closure of both defects. This shows that closing small fenestrations that are far away from the primary interatrial defect without rims and using other devices instead, such as a PDA coil, is feasible and can avoid the need for an open-heart surgical procedure; moreover, it is important to note that leaving these fenestrations open can have the same physiology as a patent foramen oval.

Improvement in edema and cognitive recovery after moderate traumatic brain injury with the neurosteroid prodrug NTS-104.

Neuroactive steroids reduce mortality, decrease edema, and improve functional outcomes in preclinical and clinical traumatic brain injury (TBI) studies. In this study, we tested the efficacy of two related novel neuroactive steroids, NTS-104 and NTS-105, in a rat model of TBI. NTS-104 is a water-soluble prodrug of NTS-105, a partial progesterone receptor agonist. To investigate the effects of NTS-104 on TBI recovery, adult male Sprague Dawley rats received moderate parasagittal fluid-percussion injury or sham surgery and were treated with vehicle or NTS-104 (10 â€‹mg/kg, intramuscularly) at 4, 10, 24, and 48 â€‹h post-TBI. The therapeutic time window was also assessed using the neuroactive steroid NTS-105 (3 â€‹mg/kg, intramuscularly). Edema in the parietal cortex and hippocampus, measured at 3 days post-injury (DPI), was reduced by NTS-104 and NTS-105. NTS-105 was effective in reducing edema when given at 4, 10, or 24 â€‹h post-injury. Sensorimotor deficits in the cylinder test at 3 DPI were ameliorated by NTS-104 and NTS-105 treatment. Cognitive recovery, assessed with cue and contextual fear conditioning and retention of the water maze task assessed subacutely 1-3 weeks post-injury, also improved with NTS-104 treatment. Cortical and hippocampal atrophy at 22 DPI did not improve, indicating that NTS-104/NTS-105 may promote post-TBI cognitive recovery by controlling edema and other processes. These results demonstrate that NTS-104/NTS-105 is a promising therapeutic approach to improve motor and cognitive recovery after moderate TBI.

Conditional lethality profiling reveals anticancer mechanisms of action and drug-nutrient interactions.

Chemical screens across hundreds of cell lines have shown that the drug sensitivities of human cancers can vary by genotype or lineage. However, most drug discovery studies have relied on culture media that poorly reflect metabolite levels in human blood. Here, we perform drug screens in traditional and Human Plasma-Like Medium (HPLM). Sets of compounds that show conditional anticancer activity span different phases of global development and include non-oncology drugs. Comparisons of the synthetic and serum-derived components that comprise typical media trace sets of conditional phenotypes to nucleotide synthesis substrates. We also characterize a unique dual mechanism for brivudine, a compound approved for antiviral use. Brivudine selectively impairs cell growth in low folate conditions by targeting two enzymes involved in one-carbon metabolism. Cataloged gene essentiality data further suggest that conditional phenotypes for other compounds are linked to off-target effects. Our findings establish general strategies for identifying drug-nutrient interactions and mechanisms of action by exploiting conditional lethality in cancer cells.

Dual PD-L1/SOX10 Immunohistochemistry Combined With Digital Imaging Enhances Stratification Accuracy of Patients With Metastatic Melanoma.

Immune checkpoint inhibitor therapy has demonstrated an overall survival benefit in patients with advanced melanoma. Though the significance of programmed death-ligand 1 (PD-L1) expression on melanoma cells as a predictive biomarker of response remains inconclusive, some reports indicate that a PD-L1 expression of <1% of tumor cells may be associated with better outcomes with dual immunotherapy. Adequate patient selection for combination therapy is critical given the higher frequency of adverse effects compared with monotherapy. Immunohistochemical (IHC) PD-L1 interpretation in tumor cells is challenging when inflammatory cells are present and cutoffs are low. We studied 36 metastatic melanoma biopsies from Immune checkpoint inhibitor-naive patients, previously stained and scored for PD-L1 IHC using the tumor proportion score (TPS). Cases were classified into 3 groups: <1%, 1% to 5%, and >5%. After de-coverslipping, SRY-related HMG-box-10 (SOX10) IHC was performed on PD-L1 IHC slides with a red chromogen, and subsequently scanned and scored by ≥2 dermatopathologists. This assessment determined that 25% of cases (9/36) had a TPS ≥ 1%, in contrast to the single IHC assay (63.8%). The majority of the 1-5% group (11/13, 84.6%) underwent a change of category to <1% TPS. In the >5% group, 60% of cases (6/10) were downgraded to <1% and 1% to 5% (4 and 2 cases, respectively). Our study suggests that PD-L1 IHC evaluation could benefit from dual PD-L1/SOX10 IHC. Dual IHC is expected to decrease the interference caused by PD-L1 expression on inflammatory cells, and digital imaging proves useful for the preservation and analysis of stains. Refining PD-L1 evaluation in metastatic melanoma may improve clinical decisions between single and combination immunotherapy, with potentially profound consequences in response and quality of life.

Premature ventricular contraction patterns associated with nonsustained ventricular tachycardia.

BACKGROUND: Occurrence of nonsustained ventricular tachycardia (NSVT) is associated with negative outcomes. It is not clear whether specific electrocardiographic characteristics of premature ventricular contractions (PVCs) are associated with the occurrence of NSVT. The aim of this study was to identify electrocardiographic patterns associated with the presence of NSVT during 24-h electrocardiographic monitoring in patients with >10 PVCs per hour. METHODS: This was a retrospective, observational, cross-sectional study. We reviewed consecutive patients who received 24-h ECG monitoring performed at a single outpatient cardiology center. Patients who received 24-h electrocardiographic monitoring, with a PVC burden ≥10 PVCs/h were included. Occurrence of NSVT during 24-h electrocardiographic monitoring was the main outcome. RESULTS: A total of 343 patients were analyzed (mean [SD] age, 69.7 [12.5] years; 177 men [51.6 %]). NSVT occurred in 72 patients who were compared with 271 patients without NSVT. The novel term "premature beat ratio", which aims to correlate the coupling interval and compensatory pause, was introduced; a value >0.5 was independently associated with NSVT according to the multivariate model (OR = 3.73, 95 % CI = 1.57-8.82; P = 0.002). PVC burden (OR = 1.09, 95 % CI = 1.02-1.17; P = 0.006), and triplets (OR = 18.19, 95 % CI = 7.32-45.18 P = 0.0) were also associated with NSVT in the multivariate model. CONCLUSION: These findings suggest that patients with a high PVC burden, triplets, and a premature beat ratio greater than 0.5 have an increased probability of presenting with NSVT and may benefit from more rigorous follow-up.

Encephalomyelitis and serositis caused by Chlamydia pecorum in buffalo calves from Brazil.

Chlamydia pecorum causes subclinical infections in cattle, but sporadic, bovine encephalomyelitis cases have been reported in calves and documented in two instances in European buffalo. An outbreak of Chlamydia pecorum-induced encephalomyelitis and serositis occurred in 3-month-old buffalo calves from Brazil. Initially presenting with pelvic limb incoordination, the calves progressed to lateral recumbency, depression, and death. Necropsies of two calves revealed encephalomyelomalacia, fibrin deposition on the external surface of the pericardium (case 1) and pleural and pericardial fibrosis (case 2). Microscopically, a multifocal to coalescing, necrotizing, neutrophilic and lymphocytic meningoencephalomyelitis with fibrinoid vasculitis and thrombosis was present. Anti-Chlamydia antibody labeling was demonstrated by immunohistochemistry. Bacteriological examination yielded no pathogenic bacteria in the brain or lungs. Chlamydia pecorum was confirmed by PCR. This work describes the gross, histopathological, microbiological, and molecular findings in two cases from an outbreak of Chlamydia pecorum-induced disease in buffalo calves.

Highly efficient generation of self-renewing trophoblast from human pluripotent stem cells.

Human pluripotent stem cells (hPSCs) represent a powerful model system to study early developmental processes. However, lineage specification into trophectoderm (TE) and trophoblast (TB) differentiation remains poorly understood, and access to well-characterized placental cells for biomedical research is limited, largely depending on fetal tissues or cancer cell lines. Here, we developed novel strategies enabling highly efficient TE specification that generates cytotrophoblast (CTB) and multinucleated syncytiotrophoblast (STB), followed by the establishment of trophoblast stem cells (TSCs) capable of differentiating into extravillous trophoblast (EVT) and STB after long-term expansion. We confirmed stepwise and controlled induction of lineage- and cell-type-specific genes consistent with developmental biology principles and benchmarked typical features of placental cells using morphological, biochemical, genomics, epigenomics, and single-cell analyses. Charting a well-defined roadmap from hPSCs to distinct placental phenotypes provides invaluable opportunities for studying early human development, infertility, and pregnancy-associated diseases.

Perspectives on Synthetic Protein Circuits in Mammalian Cells.

Mammalian synthetic biology aims to engineer cellular behaviors for therapeutic applications, such as enhancing immune cell efficacy against cancers or improving cell transplantation outcomes. Programming complex biological functions necessitates an understanding of molecular mechanisms governing cellular responses to stimuli. Traditionally, synthetic biology has focused on transcriptional circuits, but recent advances have led to the development of synthetic protein circuits, leveraging programmable binding, proteolysis, or phosphorylation to modulate protein interactions and cellular functions. These circuits offer advantages including robust performance, rapid functionality, and compact design, making them suitable for cellular engineering or gene therapies. This review outlines the post-translational toolkit, emphasizing synthetic protein components utilizing proteolysis or phosphorylation to program mammalian cell behaviors. Finally, we focus on key differences between rewiring native signaling pathways and creating orthogonal behaviors, alongside a proposed framework for translating synthetic protein circuits from tool development to pre-clinical applications in biomedicine.

Medical Mistrust on Prostate Cancer Screening: A mixed method study among African Americans, Caribbean immigrants and African immigrants.

Objectives: The contribution of medical mistrust to healthcare utilization delays has been gaining increasing attention. However, few studies have examined these associations among subgroups of Black men (African Americans, Caribbean, and African immigrants) in relation to prostate cancer (PCa). This study addresses this gap by assessing how medical mistrust affects PCa screening behavior and to further understand perceptions of medical mistrust among subgroups of Black men. Methods: This research employs a mixed-methods approach comprising two distinct phases. In Phase 1, a cross-sectional examination was conducted to evaluate the influence of medical mistrust toward healthcare organizations on prostate cancer screening among 498 Black men. In Phase 2, a qualitative investigation was undertaken to delve into the nuances of medical mistrust through six focus groups (n=51) and ten key informant interviews (n=10). Logistic regression and grounded theory methods were employed for data analysis. Results: Quantitative findings unveiled disparities in mistrust among subgroups, with Caribbean immigrants exhibiting higher levels of medical mistrust. Nevertheless, individuals with a family history of PCa showed elevated likelihoods of undergoing screening, despite mistrust. Qualitative results revealed 1) differences in reasons for medical mistrust among Black subgroups, 2) cultural perceptions which influence medical mistrust and medical care seeking, 3) lack of education in relation to PCa that contributes to medical mistrust, 4) negative past experiences and poor provider communication contribute, and 5) when PCa directly affected one's life, either personally or within the family, there was a recognized importance placed on monitoring one's risk despite mistrust. Conclusion: While medical mistrust may not significantly deter healthcare utilization among individuals with a family history or diagnosis of PCa, it underscores the variability of medical mistrust and its underlying reasons among different Black subgroups.

Blowin' in the Wind: Mapping the Dispersion of Metal(loid)s From Atacama Mining.

The Atacama Desert's naturally elevated metal(loid)s pose a unique challenge for assessing the environmental impact of mining, particularly for indigenous communities residing in these areas. This study investigates how copper mining influences the dispersion of these elements in the wind-transportable fraction (<75 µm) of surface sediments across an 80 km radius. We employed a multi-pronged approach, utilizing spatial modeling to map element distributions, exponential decay analysis to quantify concentration decline with distance, regime shift modeling to identify dispersion pattern variations, and pollution assessment to evaluate impact. Our results reveal significant mining-driven increases in surface concentrations of copper (Cu), molybdenum (Mo), and arsenic (As). Notably, within the first 20 km, concentrations peaked at 1,016 mg kg⁻1 for Cu, 31 mg kg⁻1 for Mo, and a remarkable 165 mg kg⁻1 for As. Cu and Mo displayed significant dispersion, extending up to 50 km from the source. However, As exhibited the most extensive reach, traveling up to 70 km downwind, highlighting the far-reaching ecological footprint of mining operations. Mineralogical analyses corroborated these findings, identifying mining-related minerals in surface sediments far beyond the immediate mining area. Although pollution indices based on the proposed Local Geochemical Background reveal significant contamination across the study area, establishing accurate pre-industrial baseline values is essential for a more reliable assessment. This study challenges the concept of "natural pollution" by demonstrating that human activities exacerbate baseline metal(loid)s levels. Expanding monitoring protocols is imperative to comprehensively assess the combined effects of multiple emission sources, including mining and natural processes, in safeguarding environmental and human health for future generations.

Potential uses of silkworm pupae (Bombyx mori L.) in food, feed, and other industries: a systematic review.

The increasing pressures imposed on ecosystems by the growing needs of the human population are stimulus for research into innovative and unconventional sources of raw materials for different industries. This systematic review was carried out to investigate the available literature on the possible industrial uses of silkworm (Bombyx mori L.) pupae, a residue of silk production. The review was conducted using an adapted version of PRISMA. After a screening process, 105 articles were obtained and subjected to a detailed quantitative and qualitative analysis. It was found that in the last decade there has been a significant increase in the number of papers devoted to the study of the potential use of silkworm pupae in different applications, with a significantly higher number in the last three years of the scope of this review, indicating a growing interest in the subject. From the analysis of the information collected, promising uses in human and animal food, such as fish, mammalian, poultry, swine and companion animals, as well as potential uses for the pharmaceutical industry, were identified. The evaluated research identified compounds with antioxidant activity and important contents of unsaturated fatty acids, which are related to beneficial effects on cardiovascular health, diabetes control, reduction of the risk of developing certain types of cancer and inflammatory activity, among other benefits. One of the most relevant findings is that many studies report a significant concentration of α-linolenic acid in silkworm pupae oil, which is attributed with anticancer, anti-inflammatory, antioxidant, anti-obesity and neuroprotective properties, among others.

2021 NEDI-Singapore study: an updated inventory of emergency department characteristics.

INTRODUCTION: The last national emergency department (ED) inventory was performed in 2007, and major changes in population demographics, healthcare needs and infrastructure have since occurred. We sought to obtain an updated inventory of EDs in Singapore to identify and describe changes in their characteristics and capabilities across the years. METHODS: In 2021, the National Emergency Department Inventories (NEDI) instrument was administered to the leadership of Singapore EDs. Emergency departments in Singapore are opened round the clock, have no restrictions on who can access care and are equipped to handle general medical emergencies. The questionnaire comprises 16 items across three categories: (a) general characteristics, (b) patient volume and (c) medical capabilities. RESULTS: We achieved 100% response rate from all 17 EDs - nine EDs in public hospitals and eight in private hospitals. In 2021, the EDs saw a total of 1,140,388 visits, an increase of 27% from 2007, with the median number of visits almost doubling (from 39,450 to 77,989); 41% and 59% of the EDs reported over 20% of visits arriving by ambulance and over 20% of visits resulting in inpatient admission, respectively. A clear distinction between public and private EDs across these metrics remained. Medical capabilities grew: 59% had access to a dedicated computed tomography scanner (up from 46%) and 82% had negative pressure isolation facilities (up from 54%). Overall, 41% of EDs self-assessed to be operating above their capacity. CONCLUSION: Singapore EDs have progressed in capabilities and capacity. Despite this, the increasing volume, complexity and acuity of patients are imposing strains on the emergency care system, signalling potential for systems improvement.