γδ T cell antigen receptor polyspecificity enables T cell responses to a broad range of immune challenges.

γδ T cells are essential for immune defense and modulating physiological processes. While they have the potential to recognize large numbers of antigens through somatic gene rearrangement, the antigens which trigger most γδ T cell response remain unidentified, and the role of antigen recognition in γδ T cell function is contentious. Here, we show that some γδ T cell receptors (TCRs) exhibit polyspecificity, recognizing multiple ligands of diverse molecular nature. These ligands include haptens, metabolites, neurotransmitters, posttranslational modifications, as well as peptides and proteins of microbial and host origin. Polyspecific γδ T cells are enriched among activated cells in naive mice and the responding population in infection. They express diverse TCR sequences, have different functional potentials, and include the innate-like γδ T cells, such as the major IL-17 responders in various pathological/physiological conditions. We demonstrate that encountering their antigenic microbiome metabolite maintains their homeostasis and functional response, indicating that their ability to recognize multiple ligands is essential for their function. Human γδ T cells with similar polyspecificity also respond to various immune challenges. This study demonstrates that polyspecificity is a prevalent feature of γδ T cell antigen recognition, which enables rapid and robust T cell responses to a wide range of challenges, highlighting a unique function of γδ T cells.

Structural comparison and expression function analysis of BF/C2 in Ctenopharyngodon idella and Squaliobarbus curriculus.

Ctenopharyngodon idella and Squaliobarbus curriculus, members of the Cyprinidae family and Yaroideae subfamily, have shown different levels of resistance to grass carp reo virus (GCRV), with S. curriculus exhibiting higher resilience. In the pursuit to explore the distinctions in the structural and expression traits of BF/C2 (A,B) between the two species, we conducted an analysis involving the cloning and examination of various coding sequences (CDS). We successfully cloned the CDS of ci-BF/C2A and ci-BF/C2B from C. idella, which spanned 2259 bp and 2514 bp respectively, encoding 752 and 837 amino acids. Similarly, the CDS of sc-BF/C2A and sc-BF/C2B from S. curriculus were cloned, featuring lengths of 1353 bp and 2517 bp and encoding 450 and 838 amino acids, respectively. A chromosome collinearity assessment revealed that ci-BF/C2A demonstrated collinearity with sc-BF/C2A, a finding not replicated with ci-BF/C2B and sc-BF/C2B. Delving into gene structure, we discerned that ci-BF/C2A harbored a greater number of Tryp_SPc domains compared to sc-BF/C2A. Following this, we engineered and purified six prokaryotic recombinant proteins: CI-BF/C2A, CI-BF/C2A1 (a variant resulting from the deletion of the Tryp_SPc domain of CI-BF/C2A), CI-BF/C2A2 (representing the Tryp_SPc domain of CI-BF/C2A), CI-BF/C2B, SC-BF/C2A, and SC-BF/C2B. Through serum co-incubation tests with these recombinant proteins, we established the activation of the complement marker C3 in each case. Utilizing fluorescence quantitative expression analysis, we observed ubiquitous expression of ci-BF/C2A and ci-BF/C2B across all grass carp tissues, predominantly in the liver. This pattern mirrored in S. curriculus, where sc-BF/C2A was highly expressed in the gills, and sc-BF/C2B manifested notably in the liver. Kidney cell infection experiments on both species revealed enhanced resistance to GCRV post-incubation with the recombinant proteins. Notably, cells treated with SC-BF/C2 (A, B) exhibited pronounced resilience compared to those treated with CI-BF/C2 (A, B, A1, A2). However, cells incubated with CI-BF/C2A1 and CI-BF/C2A2 showed strengthen resistance relative to cells treated with CI-BF/C2A and CI-BF/C2B. In GCRV infection trials on grass carp, ci-BF/C2A and ci-BF/C2B expressions reached a zenith on the seventh day post-infection, highlighting a distinctive functional mode in immune defense against GCRV infection orchestrated by BF/C2. The empirical data underscores the pivotal role of the Tryp_SPc domain in immune responses to GCRV infection, pinpointing its influence on ci-BF/C2A expression. Conclusively, this investigation provides a foundational understanding of the unique immune function characteristics of BF/C2 in grass carp, paving the way for further scholarly exploration in this realm.

Mechanical Fourier transform for programmable metamaterials.

Proactively programming materials toward target nonlinear mechanical behaviors is crucial to realize customizable functions for advanced devices and systems, which arouses persistent explorations for rapid and efficient inverse design strategies. Herein, we propose a "mechanical Fourier transform" strategy to program mechanical behaviors of materials by mimicking the concept of Fourier transform. In this strategy, an arbitrary target force-displacement curve is decomposed into multiple cosine curves and a constant curve, each of which is realized by a rationally designed multistable module in an array-structured metamaterial. Various target curves with distinct shapes can be rapidly programmed and reprogrammed through only amplitude modulation on the modules. Two exemplary metamaterials are demonstrated to validate the strategy with a macroscale prototype based on magnet lattice and a microscale prototype based on an etched silicon wafer. This strategy applies to a variety of scales, constituents, and structures, and paves a way for the property programming of materials.

Coronavirus disease 2019: Repeated immersion of chlorine-containing disinfectants has adverse effects on goggles.

Introduction: During COVID-19, some front-line personnel experienced varying degrees of eye discomfort due to the use of goggles repeatedly disinfected with chlorine-containing disinfectant. Methods: The eye damage information of 276 front-line personnel who used goggles in a hospital from October 1, 2021, to December 1, 2021, was collected by filling out a questionnaire. To study the effect of chlorinated disinfectants on goggles, we immersed the goggles in the same volume of water and chlorinated disinfectant buckets. We tested the light transmittance, color and texture, and airtightness of the goggles at different times (1, 3, 12, 24, 36, 48, 60, 72, 96, 120, 144, 168, 192, 216, 240, and 268 h). In addition, we detected where chlorinated disinfectant remained in the goggles by using disinfectant concentration test paper. Results: 60 (21.82%) people experienced dry eyes, stinging pain, photophobia and tearing, conjunctival congestion, eyelid redness, and swelling. After treatment or rest, the patient's ocular symptoms were significantly relieved within 3 days. With the extension of disinfection time, the light transmission of the lenses gradually decreased, and the light transmission reduced when immersion occurred at 216 h. After 72 h of disinfection, the color of the goggle frame began to change to light yellow, the texture gradually became hard and brittle, and the color became significantly darker at 268 h of disinfection. The airtightness of the goggles began to decrease after 168 h of disinfection, the airtightness decreased substantially at 268 h, and the shape changed significantly. In addition, the concentration test paper results show that the disinfection solution mainly resides in the goggle frame seam and goggles' elastic bands' bundle. Conclusions: Repeated chlorine disinfectant disinfection will reduce the effectiveness of goggles protection and damage front-line personnel's eye health.

Left bundle branch pacing on mechanical synchrony and myocardial work in bradycardia patients.

Left bundle branch pacing (LBBP) has emerged as a novel physiological pacing method to produce narrower QRS duration, but whether it could restore mechanical synchrony and improve myocardial work still lacks sufficient evidence. Therefore, the goal of this study was to evaluate mechanical synchrony and myocardial work in LBBP. We collected 20 patients with LBBP due to symptomatic bradycardia and another 29 age-matched patients with right ventricular pacing (RVP). For LBBP patients, cardiac electro-mechanical synchrony and myocardial work were measured at baseline and 7 days after implantation and compared with the RVP patients. In the LBBP group, paced QRS duration and mechanical synchrony were not significantly different from baseline(all P > 0.05), but significantly smaller than that in the RVP group (all P<0.05). Meanwhile, global longitudinal strain (GLS) in LBBP was greater than that in the RVP group (17.7 ± 3.5% vs. 14.8 ± 3.1%, P < 0.05). Global myocardial work index and global constructive work were also better than that in the RVP group(all P<0.05). Global work efficiency was 91.9 ± 3.1%, which was greater when compared with RVP (P < 0.05). LBBP provides better cardiac electro-mechanical synchrony and more effective myocardial work than that in RVP, thus improving global heart function.

Strategies to evaluate potential effector function of glycan variants: a case study of ordesekimab (AMG 714 or PRV-015).

The potential for effector functions of therapeutic antibodies, including antibody-dependent cell-mediated cytotoxicity (ADCC), is a biological activity of interest for characterization, regardless of if ADCC is an intended primary pharmacological effect. The composition of the conserved antibody Fc glycan can vary as a function of post-translational processing which may affect the binding affinity to Fc receptors, leading to a change of effector activity. Ordesekimab (AMG 714 or PRV-015), a fully human immunoglobulin G1-kappa anti-interleukin (IL)-15 monoclonal antibody, is in clinical development for celiac disease. The binding of ordesekimab to IL-15 inhibits the interaction of IL-15 with the IL-2Rß and common γ chain of the IL-15 receptor complex, but not with the IL-15Rα chain. Therefore, the simultaneous binding of ordesekimab to the Fcγ receptor (R) IIIα expressed on natural killer (NK) cells and to the IL-15/IL-15Rα complex on cells such as monocytes may theoretically enable ADCC toward the IL-15Rα-expressing cells. The high mannose (HM) levels on the Fc glycan were found to vary in different lots of ordesekimab resulting from refinements to the manufacturing process, and the impact on ordesekimab-mediated ADCC activity was evaluated in in vivo and in vitro studies. A review of nonclinical and clinical data found no evidence of ordesekimab-induced depletion of monocytes, or cytotoxicity in organs with wide IL-15Rα expression, suggesting a lack of in vivo ADCC activity. In addition, in vitro peripheral blood mononuclear cells-based ADCC assay did not reveal any cytolytic effect of ordesekimab with various levels of HM content when cocultured with recombinant human IL-15. Taken together, these data demonstrate that ADCC is not a potential liability for ordesekimab and does not contribute to the reduction of IL-15-mediated inflammation, the intended pharmacological effect.

An Organic Laser Based on Thermally Activated Delayed Fluorescence with Aggregation-Induced Emission and Local Excited State Characteristics.

The spatial separation between the highest occupied and the lowest unoccupied molecular orbitals (HOMO and LUMO) in thermally activated delayed fluorescent (TADF) molecules leads to charge transfer (CT) states, which degrade the oscillator strength of emission transition and sacrifices high solid-state photoluminescence quantum yield (PLQY), together limiting its application in organic solid-state lasers (OSSLs). Here, we demonstrated organic microwire lasers from TADF emitters that combine aggregation induced emission (AIE) and local excited (LE) state characteristics. The unique AIE and LE feature lead to a PLQY approaching 50 % and a high optical gain of 870 cm-1 for TADF microwires. The regenerated singlet excitons by reverse intersystem crossing (RISC) process are conducive to population inversion. As a result, we demonstrated microwire lasers around 465 nm with a low threshold of 3.74 µJ cm-2 . Therefore, our work provides insight to design TADF materials for OSSLs.

Characterization of an Anti-CD70 Half-Life Extended Bispecific T-Cell Engager (HLE-BiTE) and Associated On-Target Toxicity in Cynomolgus Monkeys.

Bispecific T-cell engager (BiTE) molecules have great potential to treat cancer. Nevertheless, dependent on the targeted tumor antigen, the mechanism of action that drives efficacy may also contribute to on-target/off-tumor toxicities. In this study, we characterize an anti-CD70 half-life extended BiTE molecule (termed N6P) which targets CD70, a TNF family protein detected in several cancers. First, the therapeutic potential of N6P was demonstrated using in vitro cytotoxicity assays and an orthotopic xenograft mouse study resulting in potent killing of CD70+ cancer cells. Next, in vitro characterization demonstrated specificity for CD70 and equipotent activity against human and cynomolgus monkey CD70+ cells. To understand the potential for on-target toxicity, a tissue expression analysis was performed and indicated CD70 is primarily restricted to lymphocytes in normal healthy tissues and cells. Therefore, no on-target toxicity was expected to be associated with N6P. However, in a repeat-dose toxicology study using cynomolgus monkeys, adverse N6P-mediated inflammation was identified in multiple tissues frequently involving the mesothelium and epithelium. Follow-up immunohistochemistry analysis revealed CD70 expression in mesothelial and epithelial cells in some tissues with N6P-mediated injury, but not in control tissues or those without injury. Collectively, the data indicate that for some target antigens such as CD70, BiTE molecules may exhibit activity in tissues with very low antigen expression or the antigen may be upregulated under stress enabling molecule activity. This work illustrates how a thorough understanding of expression and upregulation is needed to fully address putative liabilities associated with on-target/off-tumor activity of CD3 bispecific molecules.

Thin films composed of Zr-doped In2O3 grains rich in fracture surfaces and cracks for photoelectrochemical water oxidation.

Zr-doped In2O3 thin films are prepared on FTO substrates by a two-step method: firstly, Zr-doped In(OH)3 thin films are hydrothermally deposited, and then converted to Zr-doped In2O3 films by heat treatment. It is found that during the phase transition from Zr-doped In(OH)3 to Zr-doped In2O3, the cuboid-like crystal grains will fragment, resulting in a large number of new surfaces and cracks. Zr doping can introduce shallow impurity levels in the band gap of In2O3, which will enhance the absorption of incident light. The substitution of trivalent In3+ ions by tetravalent Zr4+ ions provides additional donors for In2O3, which reduces the charge transfer resistance of the photoelectrochemical water oxidation and thus improves the charge transfer kinetics. These factors synergistically improve the photoelectrochemical water oxidation performance of Zr-doped In2O3. For example, at a potential of 1.5 V versus reversible hydrogen electrode, the photocurrent density of the Zr-doped In2O3 electrode during photoelectrochemical water splitting can be as high as about 3.5 times that of the undoped In2O3. Furthermore, Zr doping will also cause changes in the nucleation of some In(OH)3 grains, resulting in the formation of a small number of rod-bundle-shaped grains.

Applying a Psychological Nursing Care Quality Evaluation Index in hospitalized patients: A pilot study.

Psychological problems have become a significant public health problem. Appropriate mental health care is crucial in promoting patient care quality. This study aimed to test the feasibility of a Psychological Nursing Quality Evaluation Index in hospitalized patients. This is a pilot study with patients hospitalized with myocardial infarction from July to September 2020 in China. The researchers used an observational approach to examine nurses' psychological health care performance based on the Psychological Nursing Quality Evaluation Index. The results indicated high compliance rates of nurses' psychological care performance, which provides references for evaluating and monitoring inpatient psychological nursing care.

Polarity-active NIR probes with strong two-photon absorption and ultrahigh binding affinity of insulin amyloid fibrils.

Amyloid fibrils are associated with many neurodegenerative diseases. In situ and in vivo visualization of amyloid fibrils is important for medical diagnostics and requires fluorescent probes with both excitation and emission wavelengths in the far-red and NIR region, and simultaneously with high binding-affinity to amyloid fibrils and the ability to cross the blood-brain barrier, which, however, remain a challenge. Here, we rationally design and synthesize an excellent polarity-sensitive two-photon excited NIR fluorophore (TZPI) based on a donor (D)-acceptor (A)-ion compound. The electron-rich carbazole group and the ionic pyridinium bromide group, linked by an electron-poor π-conjugated benzothiadiazole group, ensure strong near infrared (NIR) emission. Furthermore, the lipophilic carbazole together with the benzothiadiazole group facilitates docking of the probe in the hydrophobic domains of amyloid aggregates with the dissociation constant K d = 20 nM and 13.5-fold higher binding affinity to insulin fibrils than the commercial probe ThT. On association with the amyloid fibrils, the tiny decrease in polarity leads to a large increase in its NIR emission intensity with an on-off ratio > 10; meanwhile, the TZPI probe exhibits a quantum yield of up to 30% and two-photon absorption cross-section values of up to 467.6 GM at 890 nm. Moreover, the application of TZPI in two-photon imaging is investigated. The ultrahigh binding affinity, the strong NIR emission, the good two-photon absorption properties, the high photo-stability, the appropriate molecular mass of 569 Da and the lipophilicity with log P = 1.66 ± 0.1 to cross the BBB make TZPI promising as an ideal candidate for detecting amyloid plaques in vivo.

Brain transcriptome analysis reveals genes involved in parental care behaviour in discus fish (Symphysodon haraldi).

Parental care is common in mammals and allows offspring to obtain milk, a substance rich in a range of nutritional and non-nutritional factors crucial to the survival of newborns. The discus fish Symphysodon spp., an Amazonian cichlid, shows an unusual behaviour: Free-swimming fry bite on their parents' skin mucus for growth and development during the first month after hatching. This is similar to the breastfeeding behaviour of mammals, but little is known about the regulatory mechanism by which discus secrete 'milk' and the related genes involved in parental care. Here, transcriptome sequencing was performed by using the brain tissues of female discus fish in parental and non-parental care. The results showed that a total of 86 differentially expressed genes (71 up-regulated genes and 15 down-regulated genes) were obtained by comparing parental with non-parental discus fish, including up-regulated LAPTM, FOXB, SOX1S, OTX2 and NR1F2, and down-regulated EDNRB, PRKCD, H1-5 and HBE. Through functional enrichment analysis, a total of 20 pathways were identified, e.g., estrogen signaling pathway, inflammatory mediator regulation of TRP channels, vascular smooth muscle contraction, GnRH signaling pathway, neurotrophin signaling pathway, NOD-like receptor signaling pathway, Jak-STAT signaling pathway, Fc gamma R-mediated phagocytosis, serotonergic synapse, autophagy-animal and cytokine-cytokine receptor interaction. These pathways and related genes might play important roles in the regulation of discus 'milk' secretion.

One-dimensional ZnO micro/nanostructures: deep insight into the growth mechanism and fine control of the microscopic morphology.

One-dimensional zinc oxide arrays with various finely controlled microscopic morphologies are grown on fluorine-doped tin oxide substrates by a hydrothermal method. The relationship between the microscopic morphologies and the reaction conditions are studied deeply. It is found that although all the studied reaction parameters, like reaction time, pH value, concentration of the reactants and so on, can affect the microstructures of the resultant products, what they affect, in essence, is the concentration of free Zn2+ ions of the solution. By exploring the evolution of the microstructure of the one-dimensional zinc oxide crystals, it is proved that the formation of the various microscopic morphologies is a result of the competition between the kinetic control and the thermodynamic control during the crystal growth, which in turn is mainly determined by the concentration of the free Zn2+ ions in the solution. The in-depth exploration of the growth mechanism of zinc oxide and the fine control of its microscopic morphologies is expected to provide advanced materials for current and future cutting-edge applications of zinc oxide. It is also expected that the growth mechanism reported here can provide theoretical support for achieving other oxide materials whose microstructure can be finely controlled.

Molecular Design Strategy for Simultaneously Strong Luminescence and High Mobility: Multichannel CH-π Interaction.

It is a big challenge to achieve high-performance organic semiconductor materials integrating both high luminescence efficiency and carrier mobility, because they are commonly regarded as a pair of contradiction. Here, combining a tight-binding model and density functional theory/time-dependent density functional theory, we propose a theoretical protocol to characterize the luminescence efficiency via an excitonic effective mass and charge transport ability via charge effective mass at the same level. Applying this protocol to a series of organic semiconductor materials, we find that the multichannel CH-π interaction can induce a heavy excitonic effective mass and light charge effective mass, which effectively balance the light-emitting efficiency and carrier mobility. Thus, a practical molecular design strategy is figured out to exploit novel organic semiconductor materials with strong luminescence and fast carrier transport simultaneously.

Mental distress and influencing factors in nurses caring for patients with COVID-19.

BACKGROUND: Nurses are experiencing tremendous stress during the new coronavirus disease 2019 (COVID-19) pandemic, especially intensive care nurses. The pandemic of the disease is a tragedy, which may leave a catastrophic psychological imprint on nurses. Understanding nurses' mental distress can help when implementing interventions to mitigate psychological injuries to nurses. AIMS AND OBJECTIVES: To quantify the severity of nurses' post-traumatic stress disorder (PTSD) symptoms and stress and explore the influencing factors of their psychological health when caring for patients with COVID-19. DESIGN: A cross-sectional survey. METHODS: The PTSD Checklist-Civilian and the Perceived Stress Scale were administered from 11 to 18 March 2020, to 90 nurses selected from another city to go and help an intensive care unit (ICU) in Wuhan, China. These nurses were selected because of their high levels of clinical performance and resilience status. RESULTS: Nurses' average PTSD score was 24.62 ± 6.68, and five (5.6%) of the nurses reported a clinically significant level of PTSD symptoms (>38 points). Nurses' perceived stress averaged 19.33 ± 7, and 20 nurses (22.22%) scored positively >25 points. Nurses' stress and PTSD symptoms were positively correlated (P < .01). Major stress sources included working in an isolated environment, concerns about personal protective equipment shortage and usage, physical and emotional exhaustion, intensive workload, fear of being infected, and insufficient work experiences with COVID-19. CONCLUSIONS: This study showed that even relatively highly resilient nurses experienced some degree of mental distress, including PTSD symptoms and perceived stress. Our findings highlight the importance of helping nurses cultivate resilience and reduce stress. RELEVANCE TO CLINICAL PRACTICE: Recommendations for practice include providing adequate training and orientation before assigning nurses to ICU to help, offering disaster-emergency-preparedness training to keep nurses prepared, providing caring and authentic nursing leadership, offering ongoing psychological support to frontline nurses.

Potential Risks for Seahorse Stock Enhancement: Insight From the Declivity of Genetic Levels With Hatchery Management.

Stock enhancement is one of the potential management strategies for the fishery. To better understand the impaction of stock enhancement, we simulated an experiment for lined seahorse (Hippocampus erectus) and evaluated the genetic structure after stock enhancement. In this study, we found the numbers of alleles (N A ) and heterozygosity (H O ) of stock enhancement strains were lower than those of the wild collections, while the inbreeding coefficient (F IS ) and relatedness index were higher. Within the 3 generations of stock enhancement strain, the N A , H O and polymorphism information content (PIC) didn't change significantly. In addition, the F ST value indicated that the genetic differentiation between the stock enhancement strains and the first wild collection reached an intermediate level, which could lead to substructuring in wild populations. Overall, these findings revealed a potential genetic risk associated with the release of hatchery strains into wild populations.

Current State and Influencing Factors of Nurse Resilience and Perceived Job-Related Stressors.

BACKGROUND: Resilience is a characteristic and skill that nurses can learn. This study examined the current state and influencing factors of nurse resilience and nurse perceived job-related stressors. METHOD: This cross-sectional survey study was conducted at a university-affiliated hospital in China between May and August 2018. The Connor-Davidson Resilience Scale was used to measure nurse resilience. RESULTS: A total of 2,981 nurses participated in the study, with an average resilience score of 61.35 ± 13.12. Nurse resilience was significantly correlated with age, years of employment, clinical rank, and education (p < .05). Main job-related stressors included frequent inspections and examinations, heavy workload, mandatory overtime, and low wages. CONCLUSION: The participants had resilience scores that were lower than in the general public in the United States and China, as well as in nurses in developed countries. This study indicated a need for hospital leaders to find ways to reduce nurse work-related stress. Building nurse resilience should be an important focus for leaders. [J Contin Educ Nurs. 2020;51(3):132-137.].

Fluorine and tin co-doping synergistically improves the photoelectrochemical water oxidation performance of TiO2 nanorod arrays by enhancing the ultraviolet light conversion efficiency.

Fluorine and tin co-doped rutile TiO2 nanorod arrays are grown on fluorine-doped tin oxide substrates by a hydrothermal process and are used as photoanodes to perform photoelectrochemical water oxidation. Fluorine and tin co-doping synergistically enhances the ultraviolet light conversion efficiency of the resulting TiO2, which enables its photocurrent density of photoelectrochemical water oxidation to be more than four times that of the undoped samples. Such improvement in photoelectrochemical performance is attributed to changes in the electronic structure of the rutile TiO2 due to fluorine and tin co-doping. It is found that introducing tin into the matrix of rutile TiO2 can improve the charge separation efficiency because of the enhanced migration of photogenerated electrons from the conduction band of TiO2 to that of SnO2 that occurs at local sites, while fluorine doping can greatly reduce the recombination of the photogenerated electron-hole pairs due to the presence of the Ti3+ state that is produced to compensate for the charge difference between F- ions and O2- ions. It is envisaged that the fluorine and tin co-doped TiO2 nanorod arrays described will provide valuable platforms for wide photocatalytic applications that are not merely limited to photoelectrochemical water oxidation.

The association of neutrophil-to-lymphocyte ratio and delayed cerebral ischemia in patients with aneurysmal subarachnoid hemorrhage: possible involvement of cerebral blood perfusion.

Background and purpose: Emerging evidence suggests that systemic inflammation is associated with the pathophysiological process of delayed cerebral ischemia (DCI) after aneurysmal subarachnoid hemorrhage (aSAH). This study aimed to investigate the association of white blood cell (WBC) count and neutrophil-to-lymphocyte ratio (NLR) with the occurrence of DCI in SAH patients. Methods: A total of 122 patients diagnosed with aSAH within 72 h of onset were retrospectively enrolled. The count of WBC, neutrophil count (NC), and lymphocyte (LC) was collected on admission. Computed tomography perfusion was performed within 7 days after SAH. The occurrence of DCI was recorded during the hospitalization. Results: Among enrolled patients, 43 (35.2%) developed DCI during hospitalization. Patients who developed DCI had a higher count of WBC, NC, and NLR as well as a lower count of LC. NC and NLR were independently associated with the occurrence of DCI, while NLR was the best predictive parameter according to the receiver operating characteristic curve. Moreover, there was a strong correlation between NLR and mean cerebral blood flow, mean transit time and mean time to peak. Conclusion: Leukocytosis is an early pathology of SAH, and NLR may be a practical predictor for the occurrence of DCI in SAH patients.

Development of a BiTE®-mediated CD8+ cytotoxic T-lymphocyte activity assay to assess immunomodulatory potential of drug candidates in Cynomolgus macaque.

The immunotoxic potential of drug candidates is assessed through the examination of results from a variety of studies and endpoints. While the functional assessment of CD8+ cytotoxic T-lymphocytes (CTL) is well-characterized in the clinic, the lack of a robust macaque CTL functional assay has been an important hurdle in evaluating and accurately quantifying cell-mediated CD8+ T-cell effector responses in the nonclinical setting. This paper describes the development of an assay to measure CTL activity in peripheral blood mononuclear cells (PBMC) isolated from Cynomolgus macaques. A human EGFR/CD3 Bispecific T-cell Engager (BiTE®) was used to mount a robust CD8+ T-cell response in the presence of target-expressing cells. Upon target engagement, degranulation of CD107a and production of interferon (IFN)-γ both reliably indicated a robust functional response in CD8+ T-cells. The BiTE®-mediated stimulation method proved to be favorable when compared to other methods of stimulation in the absence of target cells. These studies demonstrated acceptable longitudinal variability of the functional assay and sensitivity to dexamethasone-mediated immunosuppression. Taken together, the results indicated an assay leveraging CD3-bispecific antibodies and target-expressing cells can provide a robust approach to the in vitro or ex vivo assessment of CTL function in Cynomolgus macaques. Because the impairment of CTL activity by immunomodulators is recognized to be an important contributor to decreased antiviral defense and increased carcinogenicity risk, we believe that this novel assay to be a valuable addition to the immunotoxicology assessment of therapeutic drug candidates.