Leucine Zipper-Bearing Kinase Is a Critical Regulator of Astrocyte Reactivity in the Adult Mammalian CNS.

Reactive astrocytes influence post-injury recovery, repair, and pathogenesis of the mammalian CNS. Much of the regulation of astrocyte reactivity, however, remains to be understood. Using genetic loss and gain-of-function analyses in vivo, we show that the conserved MAP3K13 (also known as leucine zipper-bearing kinase [LZK]) promotes astrocyte reactivity and glial scar formation after CNS injury. Inducible LZK gene deletion in astrocytes of adult mice reduced astrogliosis and impaired glial scar formation, resulting in increased lesion size after spinal cord injury. Conversely, LZK overexpression in astrocytes enhanced astrogliosis and reduced lesion size. Remarkably, in the absence of injury, LZK overexpression alone induced widespread astrogliosis in the CNS and upregulated astrogliosis activators pSTAT3 and SOX9. The identification of LZK as a critical cell-intrinsic regulator of astrocyte reactivity expands our understanding of the multicellular response to CNS injury and disease, with broad translational implications for neural repair

Pediatric intensive care unit treatment alters the diversity and composition of the gut microbiota and antimicrobial resistance gene expression in critically ill children

IntroductionCommon critical illnesses are a growing economic burden on healthcare worldwide. However, therapies targeting the gut microbiota for critical illnesses have not been developed on a large scale. This study aimed to investigate the changes in the characteristics of the gut microbiota in critically ill children after short-term pediatric intensive care unit (PICU) treatments.MethodsAnal swab samples were prospectively collected from March 2021 to March 2022 from children admitted to the PICU of Xinhua Hospital who received broad-spectrum antibiotics on days 1 (the D1 group) and 7 (the D7 group) of the PICU treatment. The structural and functional characteristics of the gut microbiota of critically ill children were explored using metagenomic next-generation sequencing (mNGS) technology, and a comparative analysis of samples from D1 and D7 was conducted.ResultsAfter 7 days of PICU admission, a significant decrease was noted in the richness of the gut microbiota in critically ill children, while the bacterial diversity and the community structure between groups remained stable to some extent. The relative abundance of Bacilli and Lactobacillales was significantly higher, and that of Campylobacter hominis was significantly lower in the D7 group than in the D1 group. The random forest model revealed that Prevotella coporis and Enterobacter cloacae were bacterial biomarkers between groups. LEfSe revealed that two Gene Ontology entries, GO:0071555 (cell wall organization) and GO:005508 (transmembrane transport), changed significantly after the short-term treatment in the PICU. In addition, 30 KEGG pathways were mainly related to the activity of enzymes and proteins during the processes of metabolism, DNA catabolism and repair, and substance transport. Finally, 31 antimicrobial resistance genes had significantly different levels between the D7 and D1 groups. The top 10 up-regulated genes were Erm(A), ErmX, LptD, eptB, SAT-4, tetO, adeJ, adeF, APH(3′)-IIIa, and tetM.ConclusionThe composition, gene function, and resistance genes of gut microbiota of critically ill children can change significantly after short PICU treatments. Our findings provide a substantial basis for a better understanding of the structure and function of gut microbiota and their role in critical illnesses

Visualization and Quantification of Post-Stroke Neural Connectivity and Neuroinflammation Using Serial Two-Photon Tomography in the Whole Mouse Brain

Whole-brain volumetric microscopy techniques such as serial two-photon tomography (STPT) can provide detailed information on the roles of neuroinflammation and neuroplasticity throughout the whole brain post-stroke. STPT automatically generates high-resolution images of coronal sections of the entire mouse brain that can be readily visualized in three dimensions. We developed a pipeline for whole brain image analysis that includes supervised machine learning (pixel-wise random forest models via the “ilastik” software package) followed by registration to a standardized 3-D atlas of the adult mouse brain (Common Coordinate Framework v3.0; Allen Institute for Brain Science). These procedures allow the detection of cellular fluorescent signals throughout the brain in an unbiased manner. To illustrate our imaging techniques and automated image quantification, we examined long-term post-stroke motor circuit connectivity in mice that received a motor cortex photothrombotic stroke. Two weeks post-stroke, mice received intramuscular injections of pseudorabies virus (PRV-152), a trans-synaptic retrograde herpes virus driving expression of green fluorescent protein (GFP), into the affected contralesional forelimb to label neurons in descending tracts to the forelimb musculature. Mice were sacrificed 3 weeks post-stroke. We also quantified sub-acute neuroinflammation in the post-stroke brain in a separate cohort of mice following a 60 min transient middle cerebral artery occlusion (tMCAo). Naive e450+-labeled splenic CD8+ cytotoxic T cells were intravenously injected at 7, 24, 48, and 72 h post-tMCAo. Mice were sacrificed 4 days after stroke. Detailed quantification of post-stroke neural connectivity and neuroinflammation indicates a role for remote brain regions in stroke pathology and recovery. The workflow described herein, incorporating STPT and automated quantification of fluorescently labeled features of interest, provides a framework by which one can objectively evaluate labeled neuronal or lymphocyte populations in healthy and injured brains. The results provide region-specific quantification of neural connectivity and neuroinflammation, which could be a critical tool for investigating mechanisms of not only stroke recovery, but also a wide variety of brain injuries or diseases

B Cells Migrate into Remote Brain Areas and Support Neurogenesis and Functional Recovery after Focal Stroke in Mice

Lymphocytes infiltrate the stroke core and penumbra and often exacerbate cellular injury. B cells, however, are lymphocytes that do not contribute to acute pathology but can support recovery. B cell adoptive transfer to mice reduced infarct volumes 3 and 7 d after transient middle cerebral artery occlusion (tMCAo), independent of changing immune populations in recipient mice. Testing a direct neurotrophic effect, B cells cocultured with mixed cortical cells protected neurons and maintained dendritic arborization after oxygen-glucose deprivation. Whole-brain volumetric serial two-photon tomography (STPT) and a custom-developed image analysis pipeline visualized and quantified poststroke B cell diapedesis throughout the brain, including remote areas supporting functional recovery. Stroke induced significant bilateral B cell diapedesis into remote brain regions regulating motor and cognitive functions and neurogenesis (e.g., dentate gyrus, hypothalamus, olfactory areas, cerebellum) in the whole-brain datasets. To confirm a mechanistic role for B cells in functional recovery, rituximab was given to human CD20+ (hCD20+) transgenic mice to continuously deplete hCD20+-expressing B cells following tMCAo. These mice experienced delayed motor recovery, impaired spatial memory, and increased anxiety through 8 wk poststroke compared to wild type (WT) littermates also receiving rituximab. B cell depletion reduced stroke-induced hippocampal neurogenesis and cell survival. Thus, B cell diapedesis occurred in areas remote to the infarct that mediated motor and cognitive recovery. Understanding the role of B cells in neuronal health and disease-based plasticity is critical for developing effective immune-based therapies for protection against diseases that involve recruitment of peripheral immune cells into the injured brain

The Analysis of Dynamic Changes and Prognosis of Posner–Schlossman Syndrome with Cytomegalovirus Infection and Antiviral Therapy

Purpose. To analyze how keratic precipitate (KP) morphology changes during Posner–Schlossman syndrome (PSS) prognosis and raise medication suggestions on 2% ganciclovir eye drops. Materials and Methods. Clinical retrospective cohort study in the Eye & ENT Hospital of Fudan University, Shanghai, China. The attacked eyes of 98 eligible subjects diagnosed unilateral PSS were enrolled between 2016 and 2019. All patients were treated with intraocular pressure-lowering drugs and anti-inflammatory steroids. 2% ganciclovir eye drops were given to cytomegalovirus (CMV) immunoglobulin G (IgG) correction ratio positive patients. Frequent follow-ups and examinations were performed. KP morphology was focused and categorized into coin-shaped, mutton-fat, and pigmented. Medical histories were noted. Multidimensional analysis was given. Results. Totally 47 patients in 98 achieved all-KP disappearance. Mean treatment time was (5.13 ± 3.66) weeks. Total KP disappearance was negatively correlated with mutton-fat and pigmented KPs at the first visit (P=0.020, P=0.007) and treatment time was also longer (P=0.018, P=0.014). Mean cumulative steroids dosage for 47 subjects was (159.66 ± 161.84) drops. CMV IgG correction ratio positive patients had smaller corneal endothelial cell density (P<0.005) and larger cup-to-disc ratio (P=0.017) than negative subjects. Cumulative steroid treatment time was longer in the CMV-positive group, and overall dosage was also larger. However, due to 2% ganciclovir eye drops, daily steroid dosage was lower in the CMV-positive group. Conclusions. The disappearance of mutton-fat and pigmented KPs needed longer treatment time. Paired aqueous humor and serum CMV IgG tests were recommended in PSS patients with coin-shaped KPs. 2% ganciclovir eye drops improved prognosis; and steroids dosage reduced significantly

Effects of Long-Term Antiglaucoma Eye Drops on Conjunctival Structures: An In Vivo Confocal Microscopy Study

Purpose. The study was aimed at comparing the long-term effects of different antiglaucoma eye drops on conjunctival structures using laser scanning confocal microscopy. Methods. Eighty patients diagnosed with primary open-angle glaucoma and twenty healthy volunteers were included in this study. The participants were divided into 5 groups according to the different medications. The lachrymal film break-up time, Schirmer’s I test, and Ocular Surface Disease Index Questionnaire were performed in all subjects. The confocal microscopy was used to observe the basal epithelial cell density (ECD), goblet cell density (GCD), dendritic cell density (DCD), and subepithelial collagen fiber diameter (SFD). Results. Statistically significant differences were found among the control group and the antiglaucoma therapy groups in the values of three clinical data (P<0.05). The GCD, DCD, and SFD showed significant differences in all glaucoma groups when compared to the control (P<0.001). Moreover, the prostaglandin group differed from the other antiglaucoma therapy groups in the GCD and SFD (P<0.05). Conclusions. Our study confirmed the significant differences in the conjunctival structures based on the effects of antiglaucoma medications. Less pronounced changes were found in the patients treated with prostaglandin analogue than in the other kinds of antiglaucoma therapies

Value of Central Venous Pressure Monitoring in the Patients with Sepsis-Associated Acute Kidney Injury

Background. Although the measurement of central venous pressure (CVP) is a common clinical tool, the role of CVP monitoring in the outcome of sepsis is controversial because threshold values of CVP are uncertain, and there are only limited data on short-term survival of patients with septic acute kidney injury (AKI). Methods. This retrospective cohort study was based on the Medical Information Mart for Intensive Care IV (MIMIC-IV) database (source of the training dataset). Multivariate regression analysis was performed to clarify the relation between CVP measurement and clinical outcomes, and a univariate regression model after propensity score matching was utilized to validate our findings. A mortality prediction model for septic AKI and a risk stratification scoring approach were developed, and the emergency intensive care unit (eICU) database was used for external validation. Results. Of the 9170 patients in the training set, 2446 (26.7%) underwent CVP measurement. No significant association was found between CVP monitoring and 28-day mortality among patients with septic AKI (odds ratio=0.479; 95% confidence interval 0.213-1.076, P=0.075), even after adjustments (propensity score matching; P=0.178). Length of ICU stay and hospital stay was markedly reduced in patients undergoing CVP measurement within 3 hours (median 6.2 and 10.9 days, respectively, P<0.001). The addition of the mean perfusion pressure initial, CVP, and the magnitude of the CVP change within 48 hours to the model significantly increased model discrimination (area under the receiver operating characteristic curve: 0.867 and 0.780, respectively, P<0.001). Conclusions. These findings suggest that CVP measurement alone has little effect on the outcome of septic AKI. Nonetheless, initial CVP levels and the dynamic changes in CVP within the first 48 hours after ICU admission and the mean perfusion pressure initial can improve the accuracy of outcome prediction models

The Characteristics of Peripapillary Retinal Perfusion by Optical Coherence Tomography Angiography in Tessellated Fundus Eyes

<div><p>Purpose</p><p>To evaluate the peripapillary and perifoveal retinal perfusions of young healthy eyes with a tessellated fundus using optical coherence tomography (OCT) angiography.</p><p>Methods</p><p>Thirty-five Chinese subjects with a tessellated fundus and 35 subjects without a tessellated fundus from a population-based cross-sectional study in Shanghai were included. All participants underwent OCT angiography. The flow index and vessel density were examined in the peripapillary and perifoveal retinal areas, and their relationships with other ocular parameters were analyzed.</p><p>Results</p><p>In the peripapillary area, the eyes with a tessellated fundus had a lower retinal nerve fiber layer (RNFL) flow index (0.055 ± 0.009 vs. 0.061 ± 0.007, P = 0.006), RNFL vessel density (61.8 ± 7.3 vs. 65.9 ± 5.2, P = 0.010), retinal flow index (0.086 ± 0.010 vs. 0.092 ± 0.008, P = 0.012), and retinal vessel density (83.7 ± 5.0 vs. 86.4 ± 3.7, P = 0.018) than the control eyes, and the difference remained significant even after adjustments were made for gender and RNFL thickness. No difference was found in the perifoveal area. Multivariable linear regression analysis showed that the retinal flow index and vessel density in the peripapillary area were significantly correlated with the tessellated fundus diagnosis (flow index: β = -0.006, P = 0.005; vessel density: β = -2.597, P = 0.006), gender (flow index: β = 0.005, P = 0.019; vessel density: β = 3.129, P = 0.002) and RNFL thickness (flow index: β = 0.000, P = 0.002; vessel density: β = 0.190, P = 0.002). The RNFL flow index and vessel density were significantly associated with the tessellated fundus diagnosis (flow index: β = -0.005, P = 0.005; vessel density: β = -3.572, P = 0.008) and the thickness of RNFL (flow index: β = 0.001, P < 0.001; vessel density: β = 0.421, P < 0.001).</p><p>Conclusions</p><p>Eyes with tessellated fundus with a relative decreased peripapillary retinal perfusion compared with eyes without a tessellated fundus were observed. The findings whether indicate causality that the reduction in the peripapillary perfusion and the peripapillary atrophy in myopia, need further study.</p></div

Peripapillary Retinal Perfusion and Stepwise Model Multivariable Associations.

<p>Peripapillary Retinal Perfusion and Stepwise Model Multivariable Associations.</p