Peak Levels Research Articles

Assessment of Early Intervention Strategies for Management of Cytokine Release Syndrome and Neurologic Events after Brexucabtagene Autoleucel (Brexu-Cel) Treatment in Patients with Relapsed or Refractory Mantle Cell Lymphoma (R/R MCL) in ZUMA-2

Introduction: Brexu-cel is an autologous anti-CD19 chimeric antigen receptor (CAR) T-cell therapy approved in the United States for the treatment of adults with R/R MCL and in the European Union for adults with R/R MCL after ≥2 prior therapies, including a Bruton tyrosine kinase inhibitor (BTKi). In the Phase 2 ZUMA-2 study, with a median follow-up of 35.6 months, brexu-cel showed an objective response rate of 91%, a median duration of response of 28.2 months, and a median overall survival of 46.6 months with a manageable safety profile in treated patients (pts) with R/R MCL (N=68). Grade (Gr) ≥3 infections, cytokine release syndrome (CRS), and neurologic events (NEs) occurred in 37%, 15%, and 31% of pts, respectively (Wang et al. J Clin Oncol. 2023). In August 2018, CRS and NE management strategies for ZUMA-2 were updated to initiate treatment of these adverse events earlier at the onset of Gr 1 events to improve safety outcomes. Here, we report 4-yr safety outcomes for pts in ZUMA-2 by initial and updated CRS and NE management strategies. Methods: Pts with R/R MCL and ≤5 prior treatments, including a BTKi, underwent leukapheresis and conditioning chemotherapy followed by a single infusion of brexu-cel (2×10 6 CAR T cells/kg). For CRS, the initial toxicity management strategy was to consider tocilizumab ± corticosteroids (methylprednisolone 1 mg/kg bid or dexamethasone 10 mg q6h) for Gr 2 CRS with extensive comorbidities or older age and Gr 3 CRS, and to increase the dose of corticosteroids (methylprednisolone 1 g/day x3, followed by a rapid taper consisting of 250 mg bid x2 days, 125 mg bid x2 days and then 60 mg bid x2 days) for Gr 4 CRS. The updated guidance was to administer tocilizumab for Gr 1/2 CRS that does not improve after 24 hours, and tocilizumab + corticosteroids (only if no resolution after 24 hours) and vasopressors for Gr 2 and 3 CRS. For NEs, the initial guidance was to consider use of tocilizumab for Gr 2 NEs only if other comorbid conditions exist (8 mg/kg IV over 1 hour, not to exceed 800 mg) and to add corticosteroids (dexamethasone 10 mg IV q6h, methylprednisolone 1 mg/kg bid) for Gr ≥3 NEs. In the updated guidance, steroid treatment without concurrent CRS remained the same, but both tocilizumab + dexamethasone 10 mg IV were to be administered for Gr ≥2 NEs concurrent with CRS. Both guidance protocols recommended treatment with antibiotics for management of infections. This exploratory post hoc analysis examined safety outcomes related to CRS, NEs, and infections for pts in each management group, with descriptive statistics reported. Results: As of July 23 2022, the median follow-up in the 68 treated pts was 47.5 months (range, 37.9-68.3). Of these pts, 40 received updated guidance (early intervention group [EIG]) and 28 received initial guidance (later intervention group [LIG]). Gr ≥3 CRS events were experienced at similar rates in both groups while Gr ≥3 NEs and Gr ≥3 infections occurred less often in the EIG vs the LIG (Table). Gr 5 infections occurred in 2 pts in the EIG and 0 pts in the LIG. No Gr 5 CRS or NEs occurred in either group. The median durations of Gr ≥3 CRS and infections were shorter for the EIG than the LIG while the median duration of Gr ≥3 NEs was longer for the EIG. Any Gr CRS and NEs were resolved at similar rates in both groups, but fewer any Gr infections were resolved for the EIG (all unresolved events in both groups were due to pt death before event resolution). Concomitant medications of interest were used in 80% (n=32) and 82% (n=23) of pts in the EIG and LIG, respectively. Tocilizumab was used at similar rates in the EIG and LIG (73% vs 68%), but steroids (55% vs 68%), vasopressors (18% vs 29%), and immunoglobulins (28% vs 50%) were used at lower rates in the EIG. Median CAR T-cell peak levels and area under the curve were lower for the EIG than the LIG while median CAR T-cell levels peaked at similar times in both groups. Conclusions: With an updated safety management protocol emphasizing early intervention with tocilizumab and corticosteroids for CRS and NEs, only 20% of pts had Gr ≥3 NEs and 30% had Gr ≥3 infections. Additionally, a lower percentage of EIG pts required steroid, vasopressor, and immunoglobulin use than pts in the LIG. CAR T-cell expansion levels were numerically lower in the EIG. Although limited by small pt numbers, these findings support the current clinical guidance for safety management during brexu-cel treatment, which is close to the ZUMA-2 updated toxicity management guidance that was examined in this study.

In Vivo Generation of Functional CD19 CAR-T Cells Using a Serum-Stable CD3-Targeted Lentiviral Vector

The high cost and complexity of CAR-T therapy has spurred efforts to generate CAR-T cells directly within patients via systemic infusion of serum-stable lentiviral vector particles that target T cells in situ for CAR transgene delivery. To ensure effective in vivo delivery, these vectors need to show resistance to neutralization by human serum, demonstrate transduction specificity for T cells, and support subsequent expansion of CAR-T cells. Lentiviral vectors (LV) pseudotyped with VSV G-glycoprotein (G-WT) transduce target cells via the LDL receptor (LDLR), which is ubiquitously expressed at variable levels and is upregulated on activated T cells. G-WT LVs are widely used to generate therapeutic CAR-T cells ex vivo, following purification and ex vivo activation of the T cells. To generate LVs capable of targeting circulating T cells in vivo, we utilized rational engineering of the VSV-G glycoprotein to design a G-CD3. This modified G protein avoids LDLR recognition, incorporates a CD3-specific single-chain antibody for targeted recognition of T cells, and minimizes LV particle inactivation upon exposure to human serum. G-CD3 LVs were shown to target and transduce resting T cells in peripheral blood mononuclear cell (PBMC) cultures with very high efficiency. The G-CD3 LVs powerfully activated T cells, evident from their increased CD25 expression, and the infection was not hindered in the presence of fresh, complement-active human serum (25%). Additionally, G-CD3 LVs efficiently transduced T cells in fresh human whole blood without apparent off-target transduction of non-T cell lineages. In vivo evaluation of G-CD3 LV efficiency was performed in humanized NSG mice by intravenous (IV) or intraperitoneal (IP) delivery of G-CD3 LVs encoding anti-CD19 CAR and GFP transgenes. Both IV and IP delivery of G-CD3 LV led to in vivo CAR T cell generation with peak levels of 17% of total circulating human CD45+ cells on day 21 post vector administration. The in vivo generated CD19-CAR-T cells were fully functional as evidenced by the disappearance of endogenous human CD19+ B cells from peripheral blood. Tissue analysis of spleen, bone marrow, and liver at the time of sacrifice (day 39 post LV injection), showed anti-CD19 CAR-T in all three tissues accompanied by an absence or dramatic reduction in tissue-resident human CD19+ B cells. In contrast, CAR-T cells were not detected at any time point in humanized NSG mice injected with G-WT LV, regardless of whether the virus was administered IV or IP. In summary, these data highlight the potential applicability of fully retargeted, serum stable LVs for in vivo gene delivery or genome editing, offering promising prospects for future human gene therapy applications.

A Pro-Inflammatory State and Peak Cytokines Are Associated with Toxicity and Early Responses in Real-World Multiple Myeloma Patients Treated with Idecabtagene Vicleucel

Introduction. Idecabtagene vicleucel (ide-cel) is a chimeric antigen receptor T-cell therapy (CAR-T) approved for patients with triple-class exposed relapsed/refractory multiple myeloma (TCE RRMM) (Munshi NC, et al. N Engl J Med 2021;384:705-716). Treatment-related toxicities, including cytokine release syndrome (CRS) and immune effector cell-associated neurotoxicity syndrome (ICANS) are prevalent and common to all CAR T-cell therapies. We examined the association of serum cytokines and pre-CAR-T bone marrow composition with toxicity and early response among patients with TCE RRMM treated with ide-cel. Methods. Patients treated with commercial ide-cel at H. Lee Moffitt Cancer Center between May 5 th, 2021 to May 18 th, 2023, and that consented to a biospecimen collection protocol, were included. Serum cytokine levels (GM-CSF, IL2, IL6, IL15, IFNγ, Angiopoietin 1&2 [ANG1, ANG2] and TNFα) were measured at apheresis, pre-lymphodepleting (LD) chemotherapy (day -6), infusion (D0) and at least weekly until day 28 using the Ella system (ProteinSimple). Inflammatory markers (C-reactive protein [CRP], ferritin, and albumin) at pre-LD were measured in the clinical laboratory. Marrow biopsy plasma cell burden (PCB) and marrow aspirate immune cell phenotyping of T cells, myeloid cells, and natural killer (NK) cells by flow cytometry were calculated prior to LD. High PCB was considered ≥50% CD138+ plasma cells. CRS and ICANS were graded per the American Society for Transplantation and Cellular Therapy (ASTCT) consensus criteria. Patient characteristics, cytokines, and other biomarkers were examined by any grade and ≥ grade 2 CRS and ICANS and day 90 (D90) response using Wilcoxon rank sum or chi-square tests. Results. 86 patients were included, and 30 (35%) were analyzed by flow cytometry. Patient characteristics included a median age of 66 years (range, 43-83), 30% high PCB, 33% high-risk cytogenetics, and a median of 6 prior lines of therapy (range 4-13). Any grade CRS occurred in 80% of the patients, ≥ grade 2 CRS in 19%, any grade ICANS in 14%, and ≥ grade 2 ICANS in 10%. Tocilizumab was administered to 65%, 29% received steroids, and 2% received anakinra. At D90, 52/78 (67%) had achieved ≥ partial response (PR) and 26/78 (33%) had either stable disease (SD), progressive disease (PD) or had died. Three deaths occurred by D90 and were attributed to complications of myeloma progression. Older patients (≥ 65 years) were more likely to develop any grade CRS (p=0.04) and ICANS (p=0.047), and those requiring bridging chemotherapy were more likely to develop ICANS (p=0.025). High PCB was not associated with CRS or ICANS, but was associated with T cell exhaustion and increased CD56 dim NK-cells (p<0.05, Figure A). At apheresis, patients with higher TNFα levels were more likely to develop ICANS (p=0.004), but no other cytokines were associated with toxicity at this timepoint, although patients with CRS and ICANS had numerically higher IL-6. At LD, elevated ratio of ANG2/ANG1 (p=0.029), IL-15 (p=0.019), and high CRP/low albumin ratio (p=0.007) were observed for patients that developed ICANS. Higher peak cytokines, ferritin, and CRP were associated with any grade CRS (p<0.05), and higher peak IL6 was associated with grade ≥ 2 CRS (p=0.03). Patients that developed any grade ICANS had higher peak IL6, IL15, ANG2 /ANG1 ratio, ferritin, and CRP (p<0.05), and these associations persisted for IL15, ANG2/ANG1 ratio, ferritin, and CRP for grade ≥ 2 ICANS (p<0.05). Achievement of ≥PR at D90 was associated with higher peak levels of GM-CSF, IL2, IFNγ, and TNFα (p<0.05, Figure B). Conclusion. In this analysis of real-world TCE RRMM patients treated with commercial ide-cel, inflammatory markers, pre-treatment marrow plasma cell burden, and immune cell subsets were associated with treatment-related toxicities and early responses. These findings provide insights into potential avenues for toxicity prevention and therapeutic optimization in high-risk patients. Additional studies in a larger patient cohort are warranted to understand the complex relationships between inflammation, immunity, toxicity, and efficacy among TCE RRMM patients treated with ide-cel.

T-Charge™ Manufacturing of the Anti-BCMA CAR-T, Durcabtagene Autoleucel (PHE885), Promotes Expansion and Persistence of CAR-T Cells with High TCR Repertoire Diversity

BCMA targeted CAR-T cells are an effective therapy for patients with relapsed or refractory multiple myeloma (r/r MM). However, autologous CAR-T cell products are highly heterogeneous and the functional roles of various T cell populations within these products have not been established. Stem-like memory T cells (Tscm) are a rare T cell subset which maintain high capacity for self-renewal and multipotency. Previously, we demonstrated that the T-Charge TM platform, a novel rapid manufacturing process that reduces manufacturing time to <2 days, preserves less differentiated CAR-T cells which exhibit potent anti-tumor activity and robust expansion in preclinical models (Dexiu Bu, ASH 2021). We conducted a phase 1 clinical trial in r/r MM (NCT04318327) of durcabtagene autoleucel (PHE885), a fully human product manufactured using the T-Charge platform. Here we present a detailed analysis of CAR-T cell products and subsequent expansion of CAR-T cells in 32 patients enrolled in this clinical trial at the Dana-Farber Cancer Institute. Apheresis products (APH), final products (FP) and post-infusion peripheral blood mononuclear cells (PBMC) were characterized using flow cytometry, mass cytometry (CyTOF) and TCR sequencing. We previously reported a 98% overall response rate (ORR) across all dose levels (2.5-20x10 6 CAR-T cells) and 100% ORR at doses >5x10 6 cell dose (Sperling, ASCO 2023). CAR-T cells expanded rapidly after infusion reaching median peak levels of 3,118 cells/ul (range 373 to 17,865) with a median 87.4% (range, 46.9-97.8) of CD3+ T cells expressing the CAR at a median of 14 days (range 10 to 27) after infusion. CAR-T cells persisted at high levels with transgene detectable by qPCR in 67% of patients at 6 months. Among 28 evaluable patients, 13 (43%) had >20% CAR positive T cells detectable by flow cytometry at 3 months. Phenotypic analysis of APH and FP samples showed that less-differentiated T cell subsets, including Tscm and central memory T cells were maintained in FP. CyTOF evaluation of functional markers revealed high expression of proliferation and activation markers in Tscm in FP and subsequently in CAR-T cells at the time of peak expansion in vivo. Subsequently, more differentiated CAR-T cells increased, accompanied by a decline in activation markers, while no significant changes were observed in inhibitory receptors. The proportion of Tscm in the FP positively correlated with early in vivo CAR-T cell expansion. TCR repertoire diversity and TCR clone tracking were used to characterize product manufacturing and CAR-T cells in peripheral blood after infusion. We sorted naïve/Tscm CD4 T cells, naïve/Tscm CD8 T cells, memory CD4 T cells, and memory CD8 T cells from APH and FP, and isolated CAR-positive CD4 T cells, CAR-positive CD8 T cells, CAR-negative CD4 cells, and CAR-negative CD8 T cells from post-infusion PBMC by fluorescence-activated cell sorting and ran TCR sequence from the extracted gDNA from each T cell subpopulation. Measurement of TCR diversity post-infusion revealed higher TCR repertoire diversity in CAR-T cells than in non-CAR-T cells. Notably, post-infusion CAR-T cells shared significantly more TCR clonotypes with T SCM than with memory T cells in the FP, suggesting that the highly heterogeneous post-infusion CAR-T clones were preferentially derived from T SCM clones in the FP. In three evaluable patients with long-term persistence, a highly diverse TCR repertoire in CAR T cells was maintained 1 year after treatment. Our findings demonstrate that the T-Charge™ manufacturing platform successfully maintains highly heterogeneous transduced Tscm clones with self-renewal potential in durcabtagene autoleucel products. Maintenance of Tscm in manufactured products contributes to robust CAR-T expansion and long-term persistence of CAR-T cells with a highly diverse TCR repertoire after infusion.

CAR-T Cell Therapy in Patients with Refractory Systemic Autoimmune Diseases Exhibits Less Inflammation, Toxicities and Different Cellular Dynamics Compared to Patients with B Cell Lymphoma

Chimeric antigen receptor (CAR) T-cell therapies have shown high efficacy in the treatment of B cell non-Hodgkin lymphoma (NHL) and acute lymphoblastic leukemia (ALL) and are established as a standard treatment for patients with hematological malignancies. Aside from malignant B cells, autoreactive B cells can also cause great harm in patients with autoimmune diseases (AID), resulting in chronic inflammation, organ damage, and increased mortality. We have recently demonstrated that CD19-directed CAR-T cell therapy is feasible and effective in patients with refractory systemic lupus erythematosus (Mougiakakos et al, 2021; Mackensen et al., 2022), antisynthetase syndrome (Müller et al., 2023) and systemic sclerosis (Bergmann et al., 2023). Despite these promising data, there are many uncertainties in the treatment of AID patients with cellular therapies, which were primarily developed for the treatment of cancer patients. Here, we analyzed the occurrence and severity of toxicities, inflammatory markers, and the cellular dynamics of 15 AID patients and 40 NHL patients undergoing CAR-T cell therapy. All patients were treated in our center with either commercial CAR-T cell products (Axicabtagenee ciloleucel, Tisagenlecleucel, Lisocabtagen maraleucel) or with the investigational medicinal product MB-CART19.1 (in-house manufactured 2 nd generation anti-CD19 CAR-T cells with a 4.1BB co-stimulatory domain; Kretschmann et al., 2023). Average peak expansion was reached at day 7 in both patient groups. Despite comparable average peak levels, patients with AID showed lower incidence and severity of cytokine-release syndrome (CRS) and immune-effector cell associated neurotoxicity syndrome (ICANS) compared to the NHL cohort. Consistent with reduced toxicity, AID patients received less tocilizumab and glucocorticoid treatment. Bone marrow toxicity with severe and/or prolonged cytopenia is a frequent long-term CAR-T cell related adverse event. Of interest, AID patients demonstrated a less profound decrease of neutrophils, platelet counts, and hemoglobin after CAR-T cell therapy compared to NHL patients and a faster recovery to normal values. Occurrence and duration of severe neutropenia (<500 neutrophils/µl) also differed between both patient cohorts. There was no late bone marrow toxicity in AID patients, whereas 19 of 40 patients with NHL showed at least one > grade 3 blood count reduction. In line with the Hematotox-Score predicting bone marrow toxicity, baseline inflammation (e.g. C-reactive protein, ferritin) prior to CAR-T cell treatment was higher in lymphoma patients than in AID patients. While AID patients quickly returned to normal ferritin values, elevated ferritin levels often persisted in the NHL cohort after CAR-T cell therapy. Given that AID patients showed fewer late toxicity and lower long-term inflammation, we speculated that persistence of CAR-T cells was lower. In line, longitudinal monitoring of the patient's immune cells including CAR-T cell kinetics demonstrated a typical contraction of CAR-T cell numbers with low-level persistence in both cohorts. We did not observe a difference in absolute CAR-T cell numbers or dynamics within the first three months after treatment. In contrast to NHL patients in remission, AID patients did not develop long-term persistence of CAR-T cells as most patients lost CAR-T cells within 1 year after cell infusion. Consistent with this observation, all AID patients showed B-cell reconstitution after loss of CAR-T cell persistence. Taken together, we observed less toxicities and severe adverse events in AID patients than in NHL patients despite similar CAR-T cell expansion and dynamics. The lack of CAR-T cell persistence and the early recurrence of B cells in AID patients is so far not explained and could provide novel insights in the biological processes controlling long-term CAR-T cell persistence.

Integrating single-cell imaging and RNA sequencing datasets links differentiation and morphogenetic dynamics of human pancreatic endocrine progenitors.

Basic helix-loop-helix genes, particularly proneural genes, are well-described triggers of cell differentiation, yet information on their dynamics is limited, notably in human development. Here, we focus on Neurogenin 3 (NEUROG3), which is crucial for pancreatic endocrine lineage initiation. By monitoring both NEUROG3 gene expression and protein in single cells using a knockin dual reporter in 2D and 3D models of human pancreas development, we show an approximately 2-fold slower expression of human NEUROG3 than that of the mouse. We observe heterogeneous peak levels of NEUROG3 expression and reveal through long-term live imaging that both low and high NEUROG3 peak levels can trigger differentiation into hormone-expressing cells. Based on fluorescence intensity, we statistically integrate single-cell transcriptome with dynamic behaviors of live cells and propose a data-mapping methodology applicable to other contexts. Using this methodology, we identify a role for KLK12 in motility at the onset of NEUROG3 expression.

Sirt3 Deletion Increases Inflammation and Mortality in Polymicrobial Sepsis.

Background: Sirtuin 3 (SIRT3) is a nicotinamide adenine dinucleotide (NAD)-dependent deacetylase that confers resilience to cellular stress by promoting mitochondrial activity. Mitochondrial dysfunction is a major driver of inflammation during sepsis. We hypothesize that Sirt3 expression improves survival in polymicrobial sepsis by mitigating the inflammatory response. Materials and Methods: Sirt3 knockout (S3KO) and wild-type (WT) mice underwent cecal ligation and puncture (CLP) or sham surgery. mRNA expression was quantified using quantitative polymerase chain reaction (qPCR) and protein expression was quantified using enzyme-linked immunosorbent assay (ELISA). Spectrophotometric assays were used to quantify serum markers of organ dysfunction. For in vitro studies, bone marrow-derived macrophages (BMDMs) were harvested from S3KO and WT mice and treated with lipopolysaccharide (LPS). Results: After CLP, hepatic Sirt3 levels decreased from baseline by nine hours and remained depressed at 24 hours. Peak serum interleukin-6 (IL-6) protein levels were higher in S3KO mice. In LPS-treated BMDMs, IL-6 mRNA levels peaked earlier in S3KO cells, although peak levels were comparable to WT. Although S3KO mice had decreased median survival after CLP compared with WT, there was no difference in five-day survival or organ dysfunction. Conclusions: Although S3KO mice initially had increased inflammation and mortality, this difference abated with time, and overall survival was comparable between the groups. This pattern is consistent with the timeline of sepsis-induced Sirt3 downregulation in WT mice, and suggests that Sirt3 downregulation occurring in sepsis is at least partially responsible for the initial hyperinflammatory response and subsequent mortality. Our data support upregulation of Sirt3 as a promising therapeutic strategy for further research in sepsis.

REVISE: Re-Evaluating the Inhibition of Stress Erosions in the ICU: a randomised trial protocol

IntroductionThe Re-Evaluating the Inhibition of Stress Erosions (REVISE) Trial aims to determine the impact of the proton pump inhibitor pantoprazole compared with placebo on clinically important upper gastrointestinal (GI) bleeding...

Small RCS targets detection based on pulse compression radar signal generator and processor

In the last two decades, field programmable gate arrays (FPGAs) have become the leading technology for implementing real time systems, specially radars. The most important advantage of FPGA is its parallel nature of executing complicated algorithms. As a result, the received data flow will be processed in a real time manner, which is suitable for radar applications. This paper attempts to show the design and the FPGA implementation of a binary phase-coded pulse compression radar signal generator and processor, that are intended to determine the ranges, and the speeds of targets with small radar cross-section area like drones. The system operating waveform is a bi-phase sequence, that is known to have the best known peak sidelobe level compared to compound Barker codes of the same length. In addition, the design incorporates a range sidelobes cancellation method to remove the time sidelobes in the matched filter output. Also, we are going to represent and investigate the FPGA implementing steps of the radar waveform generator and processor according to the required parameters. Finally, the implementation results of the proposed system will be represented.

Fish chorus variation in a tropical estuarine environmenta).

Fish choruses are still understudied in the Southwestern Atlantic Ocean. Temporal and spatial variation of fish choruses at two sites inside Guanabara Bay were investigated between 2021 and 2022; one sampling site was in a Marine Protected Area (MPA), and the other was in a rocky environment closer to vessel trafficking areas. Acoustic recordings were performed on 17 sampling occasions of 24 h, coupled to a temperature data logger. Long-term spectral averages were employed to determine choruses' start, end, and peak times, and third-octave levels were used to characterize spectral characteristics. Fish sounds were also analyzed and investigated with a principal components analysis. Choruses in the MPA lasted, on average, 4.5 h and had a peak frequency of 547.2 ± 226.6 kHz with a peak level of 104.6 ± 8.7 dB re 1 μPa. In contrast, the rocky site choruses lasted 5.5 h on average and had a peak frequency of 371.7 ± 131.0 Hz with a peak level of 113.4 ± 4.0 dB re 1 μPa. Chorus peak frequency was positively correlated to temperature (r = 0.4). Different types of fish sounds were identified, with some acoustics parameters varying between sites. Results indicate more than one chorusing species that may react to different factors.

Identification of brain areas in mice with peak neural activity across the acute and persistent phases of post-traumatic headache.

Post-traumatic headache is very common after a mild traumatic brain injury. Post-traumatic headache may persist for months to years after an injury in a substantial proportion of people. The pathophysiology underlying post-traumatic headache remains unknown but is likely distinct from other headache disorders. Identification of brain areas activated in acute and persistent phases of post-traumatic headache can provide insights into the underlying circuits mediating headache pain. We used an animal model of mild traumatic brain injury-induced post-traumatic headache and c-fos immunohistochemistry to identify brain regions with peak activity levels across the acute and persistent phases of post-traumatic headache. Male and female C57BL/6 J mice were briefly anesthetized and subjected to a sham procedure or a weight drop closed-head mild traumatic brain injury . Cutaneous allodynia was assessed in the periorbital and hindpaw regions using von Frey filaments. Immunohistochemical c-fos based neural activity mapping was then performed on sections from whole brain across the development of post-traumatic headache (i.e. peak of the acute phase at 2 days post- mild traumatic brain injury), start of the persistent phase (i.e. >14 days post-mild traumatic brain injury) or after provocation with stress (bright light). Brain areas with consistent and peak levels of c-fos expression across mild traumatic brain injury induced post-traumatic headache were identified and included for further analysis. Following mild traumatic brain injury, periorbital and hindpaw allodynia was observed in both male and female mice. This allodynia was transient and subsided within the first 14 days post-mild traumatic brain injury and is representative of acute post-traumatic headache. After this acute post-traumatic headache phase, exposure of mild traumatic brain injury mice to a bright light stress reinstated periorbital and hindpaw allodynia for several hours - indicative of the development of persistent post-traumatic headache. Acute post-traumatic headache was coincident with an increase in neuronal c-fos labeling in the spinal nucleus of the trigeminal caudalis, primary somatosensory cortex, and the nucleus accumbens. Neuronal activation returned to baseline levels by the persistent post-traumatic headache phase in the spinal nucleus of the trigeminal caudalis and primary somatosensory cortex but remained elevated in the nucleus accumbens. In the persistent post-traumatic headache phase, coincident with allodynia observed following bright light stress, we observed bright light stress-induced c-fos neural activation in the spinal nucleus of the trigeminal caudalis, primary somatosensory cortex, and nucleus accumbens. Examination of mild traumatic brain injury-induced changes in peak c-fos expression revealed brain regions with significantly increased neural activity across the acute and persistent phases of post-traumatic headache. Our findings suggest mild traumatic brain injury-induced post-traumatic headache produces neural activation along pain relevant pathways at time-points matching post-traumatic headache-like pain behaviors. These observations suggest that the spinal nucleus of the trigeminal caudalis, primary somatosensory cortex, and nucleus accumbens may contribute to both the induction and maintenance of post-traumatic headache.

Variation of Ionospheric Shortwave Absorption of the Extraordinary Wave

AbstractIn this study, associating with the A‐H formulation, we have developed a comprehensive ionospheric absorption model by incorporating the latest version of geomagnetic model, the atmospheric and ionospheric model as the background. Based on the absorption model, the ionospheric stratification theory is utilized to explore the trend of the total absorption for extraordinary (X) wave with frequency and elevation angle. We found that the factor of local time has a greater impact on the trend of total absorption variation than other factors—solar activity, season, and latitude, attributing to the variation of electron collision frequency. During the two different local times (daytime and night), the trends of the total absorption have distinct differences. In the daytime, it is worth noting that the reflection heights and the peaks of the absorption coefficient both mainly occur in Layers D and E. Therefore, the non‐deviative absorption dominates the variation of the total absorption. On the other hand, the occurrence of Pedersen ray results in inflexion points arising in the curves of absorption versus frequency or elevation angle. Another interesting phenomenon is the dominant role of deviative absorption at night. At night, the reflection heights mainly occur in Layer F, and the absorption coefficient of Layer F begins to exceed the peak levels of Layers D and E when the frequency or elevation angle is larger than certain values. Hence, the total absorption will grow as a result of the increase in deviative absorption with the increase in reflection height and absorption coefficient.

Study of temporal changes in the hydrographic network of small mountain rivers in the Ile Alatau, Kazakhstan

Abstract The article presents the outcomes of an assessment of hydrographic network changes within the Almaty city, utilizing geographic information system (GIS) technology and Earth remote sensing data. Two gauge stations were selected along the main rivers within the Almaty city. To identify distinctive alterations in these rivers, hydrological data series encompassing the maximum runoff from 1970 to 2021 were collected and subjected to statistical analysis. Differential integral curves were constructed to pinpoint periods corresponding to peak and trough runoff levels. For each of these identified periods, the processing of satellite imagery allowed for the computation of meandering coefficients for the river channels. Additionally, refinements were made to the slope values of the rivers during these same timeframes, and connectivity graphs were established to elucidate the relationship between slope and liquid runoff for each period. The analysis encompassed an assessment of the impact of anthropogenic factors on both artificial and natural bodies of water while also considering shifts in the boundaries of the Almaty city. The findings derived from this study have practical applications in the planning and design of water supply systems and in the implementation of measures aimed at mitigating the adverse effects of anthropogenic factors on water bodies.

Impact of Swirling Flow on the Overall Cooling Effectiveness and TBCs Insulation Characteristics of Turbine Vane

Modern gas turbines find extensive applications in aero engines, power generation, marine, and various other industries. Most numerical studies concentrate on turbine aero-thermal performance under different external conditions with vane or coupon; there are a few published results on combustor-turbine interactions. This study reveals the cooling performance of the first stage with or without coatings to provide a reference for high-performance combustors and turbine-integrated design. The results of the study show that (1) The velocity and temperature distribution inside the combustion chamber are obviously affected by the swirling flow. A central recirculation zone is formed near the central axis, and two external recirculation zones are formed between the inlet section of the combustion chamber and the fluid reattachment point. (2) Inside the combustion chamber, the flame temperature in the central recirculation zone is relatively high, and the range of the high-temperature zone expands with the increase of axial distance. Increasing the swirl number decreases the peak temperature level in the combustion chamber. (3) Under the influence of swirl number, the greater the swirl number, the greater the cooling effectiveness of most areas on the vane surface. (4) In regions where there is a decrease in local heat flux, positive values are evident. This suggests that the application of a coating in these areas results in a reduction of heat transfer from the vane to the mainstream. (5) When comparing the coated vane to the uncoated vane, the cooling effectiveness across the entire surface is notably enhanced, with a particularly significant improvement observed on the vane’s suction side. With the increase of Cax, the difference in cooling effectiveness increment under different swirl numbers also increases.

The Effect of Glycemic Control on Morbidity and Mortality in Critically Ill COVID-19 Patients.

Background COVID-19 infection has caused a global pandemic affecting a group of patients with chronic conditions including diabetes with exacerbating insulin resistance and hyperglycemia. Investigators noted that pre-existing diabetes and newly diagnosed diabetes are associated with an increased risk of all-cause mortality in hospitalized patients with COVID-19 infection. Aim To evaluate the relationship between ICU patients infected with COVID-19 and mortality among those with high versus low glucose levels. Methods This is a retrospective study of critically ill adult patients infected with COVID-19 who were admitted to the ICU from April 5, 2020, to October 14, 2020. The participants were from San Bernardino County which is a diverse and underserved community. Overall, 84 patients were included in the final analysis. The average age was 59.67 (standard deviation=15.55) with 59.5% being males. Overall mortality was 44.1%. Results Around one-fifth of patients had glucose under control as measured by peak glucose level of <180 mg/dL during hospital stay. A statistically significant association was seen between tighter serum glucose control and mortality (p=0.0354). Patients with serum glucose maintained <180 mg/dL were associated with significantly lower mortality than their counterparts (22.2% vs. 50%). Conclusions This study suggests that maintaining a tighter control of the glycemic index in critically ill COVID-19 patients will improve morbidity and mortality.

The utility of a baseline morning cortisol measurement, in the screening for adrenal insufficiency amongst non-critically ill hospital inpatients

BackgroundSerum cortisol levels vary according to the daily circadian cycle, with peak levels seen in the morning. There is evidence in ambulatory patients that a morning cortisol level can predict an adequate cortisol response to Synacthen (tetracosactide) stimulation. AimThe aim of this study is to determine the utility of baseline analysis of the serum cortisol levels in the morning “baseline morning cortisol”, determined using a newer immunoassay, in the screening for adrenal insufficiency amongst non-critically ill hospital inpatients. MethodsThis is a retrospective cross-sectional cohort study. Results64 adult inpatients had undergone a short Synacthen test (SST) (measurement of serum cortisol levels 30 and 60 min after administration of tetracosactide) during the study period. 17 patients returned an abnormal SST result. The measured level of cortisol in the morning correlated to both the 30 min and 60 min stimulated cortisol values (r = 0.612, p < 0.001 and r = 0.639, p < 0.001). After inspecting the receiver operating characteristic curve, a cortisol concentration of 200nmmol/L measured in the morning was selected as a threshold for predicting the SST result. Using this cut off, sensitivity for predicting a normal SST was 100 %, specificity 56 %, positive predictive value 45 % and negative predictive value 100 %. ConclusionA morning cortisol measurement, determined using a newer immunoassay, is sufficient in most cases to screen for adrenal insufficiency amongst non-critically ill hospital inpatients. Use of clinical judgement in conjunction with single morning cortisol measurement is likely to reduce the need for SST testing amongst inpatients.

Return and volatility connectedness and net directional patterns in spillover transmissions: East and Southeast Asian equity markets

AbstractIn this article, we investigate the pattern and dynamics of return and volatility connectedness across East and Southeast Asian markets (referred to as the ASEAN5 + 5 group) by utilizing forecast‐error variance decompositions in a generalized VAR framework in conjunction with the Bai‐Perron procedure to control for structural breaks. Our analysis of the dynamics of return spillovers in static and time‐varying settings identifies that the stock markets of Singapore, Hong Kong and South Korea act as constant and largest net transmitters of shocks throughout the period from January 2003 to July 2021. The Chinese stock market is found to have the lowest return connectedness with other regional markets, which could be due to the local foreign ownership regulations. Visualization of the net pairwise return spillover network shows that Singapore is the sole net transmitter of shocks to all other markets in the ASEAN5 + 5 group, whereas, China, despite its market size is the sole net recipient. Two other markets in the regional group are identified as the net receivers, Japan and the Philippines, with the former becoming a net recipient from 2007. Our analysis of structural breaks shows that return spillovers across the markets intensify during periods of economic turmoil, financial shocks and the health crisis (COVID‐19), however, return to the pre‐shock levels during stable market periods. Further analysis of time‐varying patterns revealed that the dynamic connectedness across the region is not symmetrical and the influence of negative returns is more pronounced. The investigation of volatility spillovers shows no substantial differences. The stock markets generally retain their roles. Importantly, the time‐varying volatility connectedness exhibits similar patterns and tends to reach peak levels during turbulent episodes.

Low-cost ZIF-67-modified fabrics with effective photothermal disinfection for antimicrobial personal protective equipment production

Aerosol-based transmission contributes to infection by various pathogens. Consequently, it is urgent to design antimicrobial personal protective equipment (PPE) and address the potential biological contamination of discarded PPE. We report a novel ZIF-67-modified PP fabric for photothermal disinfection; our work provides a fast, one-step, and low-cost preparation approach for incorporating zeolitic imidazolate framework (ZIF) nanoparticles (NPs) onto polypropylene (PP) fabric. First, we prepared ZIF-67-water NPs and ZIF-67-methanol NPs using relevant solvents and characterized them by X-ray diffraction, Fourier transform infrared spectroscopy, Raman spectroscopy, and field-emission scanning electron microscopy. We then used only ultrasonication to prepare ZIF-67-modified fabrics by incorporating ZIF-67 NPs onto PP fabric with a 0.5－1.5 mg/cm2 loading rate (18.5－40.5 wt%). The ZIF-67-modified fabrics were subjected to the same characterization, and then they were evaluated for photothermal performance, hydrophobicity and antimicrobial ability. The fabric of ZIF-67-water@PP-1.5 was selected to repurpose an antimicrobial mask. We demonstrated that the ZIF-67-modified fabrics have excellent photothermal performance with near-infrared (NIR) radiation and exhibit increased hydrophobicity, which is determined by the loading rate of ZIF-67 NPs. The surface temperature reached and maintained peak levels (70 °C and 90 °C) with NIR radiation, whilst the highest contact angle was over 140°. The photothermal behaviour could kill over 99.9% of gram-negative and gram-positive drug-resistant bacteria, whilst the haemolytic behaviour was less than 5%. The repurposed mask retained controllable photothermal properties. The application of these fabrics not only addresses the demand for affordable antimicrobial PPE but also contributes to reducing potential biochemical threats in discarded PPE.

Analyzing Mediating Factors in Job Satisfaction Within the Retail Superstore Environment

Employee devotion to the organization is a key factor in determining whether an organization will succeed in the competitive market. Employee job satisfaction is therefore a critical factor. Employee satisfaction leads to full commitment and motivation to perform at their peak levels, enhancing customer value and advancing business goals. When employees receive enough compensation (salary, bonus, and provident fund), allowances, and insurance, as well as better working conditions, recognition for their efforts in the workplace, opportunities for training and development, and vacation benefits from their employer, they are satisfied with their jobs. The goal of the study is to identify the factors that influence an organization's employee job satisfaction. The article's objective is to quantify the degree to which the mediating factors have an impact on worker job satisfaction.

A double-negative feedback loop between NtrBC and a small RNA rewires nitrogen metabolism in legume symbionts.

Root nodule endosymbioses between diazotrophic rhizobia and legumes provide the largest input of combined N to the biosphere, thus representing an alternative to harmful chemical fertilizers for sustainable crop production. Rhizobia have evolved intricate strategies to coordinate N assimilation for their own benefit with N2 fixation to sustain plant growth. The rhizobial N status is transduced by the NtrBC two-component system, the seemingly ubiquitous form of N signal transduction in Proteobacteria. Here, we show that the regulatory sRNA NfeR1 (nodule formation efficiency RNA) of the alfalfa symbiont Sinorhizobium meliloti is transcribed from a complex promoter repressed by NtrC in a N-dependent manner and feedback silences ntrBC by complementary base-pairing. These findings unveil a more prominent role of NtrC as a transcriptional repressor than hitherto anticipated and a novel RNA-based mechanism for NtrBC regulation. The NtrBC-NfeR1 double-negative feedback loop accurately rewires symbiotic S. meliloti N metabolism and is likely conserved in α-rhizobia.