A PACAP-activated network for secretion requires coordination of Ca2+ influx and Ca2+ mobilization.

Chromaffin cells of the adrenal medulla transduce sympathetic nerve activity into stress hormone secretion. The two neurotransmitters principally responsible for coupling cell stimulation to secretion are acetylcholine and pituitary adenylate activating polypeptide (PACAP). In contrast to acetylcholine, PACAP evokes a persistent secretory response from chromaffin cells. However, the mechanisms by which PACAP acts are poorly understood. Here, it is shown that PACAP induces sustained increases in cytosolic Ca2+ which are disrupted when Ca2+ influx through L-type channels is blocked or internal Ca2+ stores are depleted. PACAP liberates stored Ca2+ via inositol trisphosphate receptors (IP3Rs) on the endoplasmic reticulum (ER), thereby functionally coupling Ca2+ mobilization to Ca2+ influx and supporting Ca2+-induced Ca2+-release. These Ca2+ influx and mobilization pathways are unified by an absolute dependence on phospholipase C epsilon (PLCε) activity. Thus, the persistent secretory response that is a defining feature of PACAP activity, in situ, is regulated by a signaling network that promotes sustained elevations in intracellular Ca2+ through multiple pathways.

The color/shape/flavor of yam gel with double emulsified microcapsules changed synchronously in 4D printing induced by microwave.

The feasibility of 3D printing the color, aroma and shape changes of yam gel with microwave heating as stimulus and soybean protein isolate-chitosan-maltodextrin complex coacervated microcapsules rich in water-soluble betacyanin and rose essence as stimulus-response materials was discussed. The morphology of microcapsules presented irregular spherical structure, and the surface was relatively smooth and slightly concave. Microwave heating led to the gradual destruction of microcapsules in yam gel, releasing internal pigments and essence, and enhancing the redness and flavor of printed samples. The release of the water phase and oil phase of the microcapsules and the hot-spot expansion effect of the models made the 3D printed bird models bend and deform, realizing the deformation effect of "spreading of wings", which realized a three-response synchronous change in color, shape, and flavor of the printed samples within 45 s. In this study, a variety of 4D printed foods with synchronous changes in sensory characteristics were created, which increased sensory enjoyment on the basis of ensuring the nutritional needs of food.

A PACAP-activated network for secretion requires coordination of Ca 2+ influx and Ca 2+ mobilization.

Chromaffin cells of the adrenal medulla transduce sympathetic nerve activity into stress hormone secretion. The two neurotransmitters principally responsible for coupling cell stimulation to secretion are acetylcholine and pituitary adenylate activating polypeptide (PACAP). In contrast to acetylcholine, PACAP evokes a persistent secretory response from chromaffin cells. However, the mechanisms by which PACAP acts are poorly understood. Here, it is shown that PACAP induces sustained increases in cytosolic Ca 2+ which are disrupted when Ca 2+ influx through L-type channels is blocked or internal Ca 2+ stores are depleted. PACAP liberates stored Ca 2+ via inositol trisphosphate receptors (IP3Rs) on the endoplasmic reticulum (ER), thereby functionally coupling Ca 2+ mobilization to Ca 2+ influx and supporting Ca 2+ -induced Ca 2+ -release. These Ca 2+ influx and mobilization pathways are unified by an absolute dependence on phospholipase C epsilon (PLCε) activity. Thus, the persistent secretory response that is a defining feature of PACAP activity, in situ , is regulated by a signaling network that promotes sustained elevations in intracellular Ca 2+ through multiple pathways.

Sjogren's Syndrome Complicated With Aldosterone-Producing Adenoma: A Case Report.

Hypokalemia may be present in some patients with Sjogren's syndrome. When a patient with Sjogren's syndrome presents with hypokalemia, we would first consider it to be a result of the renal involvement of Sjogren's syndrome. However, in this case report, we present a young woman with Sjogren's syndrome who presented with hypokalemia that was not caused by renal tubular acidosis but by the presence of a coexisting aldosterone-producing adenoma. Cases of Sjogren's syndrome coexisting with aldosterone-producing adenoma are extremely rare. This finding underscores the need for more careful differential diagnosis in patients with Sjogren's syndrome who also have hypokalemia.

Microwave-Induced Rapid Shape Change of 4D Printed Vegetable-Based Food.

Microwave heating acts as an environmental stimulus factor to induce rapid shape changes in 4D-printed stereoscopic models over time. The influence of microwave power and model structure on the shape change behavior was explored, and the applicability of the deformed method to other vegetable-based gels was verified. The results described that the G', G″, η, and proportion of bound water of yam gels increased with the increase in yam powder content, and the yam gel with 40% content had the best printing effect. The IR thermal maps showed the microwaves first gathered in the designed gully region caused the swelling phenomenon, which induced the printed sample to undergo a bird-inspired "spreading of wings" process within 30 s. Increasing the microwave power and microwave heating time were able to increase the bending angles and dehydration rates of the printed samples, thus improving the deformed degree and deformed speed. Different model base thicknesses (4, 6, 8, and 10 mm) also had significant effects on the shape change of the printed structures. The efficiency of the shape changes of 4D-printed structures under microwave induction can be judged by studying the dielectric properties of the materials. In addition, the deformed behaviors of other vegetable gels (pumpkin and spinach) verified the applicability of the 4D deformed method. This study aimed to create 4D-printed food with personalized and rapid shape change behavior, providing a basis for the application scenarios of 4D-printed food.

Role of CD19+ CD5+ CD1d+ Bregs in maintaining the Th17/Treg balance in mice with systemic lupus erythematosus complicated with atherosclerosis.

OBJECTIVE: In this study, we aimed to investigate Bregs, their regulatory effects on Th17/Treg cell balance, and the release of downstream inflammatory factors in a mouse model of low-density lipoprotein receptor (LDLr)-/- + Pristane. METHODS: After the establishment of the mouse model of systemic lupus erythematosus (SLE) complicated with atherosclerosis (AS), 8-week-old LDLr-/- + Pristane mice (n = 10) were included in the SLE + AS group. Furthermore, 8-week-old MRL/lpr and C57 mice were used as the SLE and normal control groups, respectively (n = 10 per group). After feeding the mice a high-fat diet for 14 weeks, peripheral blood and spleen of mice were collected, and Bregs, Th17, and Treg cells and related inflammatory factors were detected by flow cytometry, enzyme-linked immunosorbent assay, and reverse-transcription polymerase chain reaction. RESULTS: The number of Bregs and Tregs in spleen lymphocytes of SLE + AS mice significantly decreased compared with the C57 group (p < .05), whereas the number of Th17 cells significantly increased (p = .000). Furthermore, the proportion of Bregs showed a negative correlation with the Th17/Treg ratio (p = .03). Mice in the SLE + AS group showed higher serum interleukin (IL)-10, IL-17, and tumor necrosis factor-α levels than those in the SLE and C57 groups (p < .05). Furthermore, IL-35 and transforming growth factor (TGF)-ß expression was reduced in the SLE + AS group compared with the C57 group (p < .05). CONCLUSIONS: The proportion of Breg decreases was negatively associated with increased Th17/Treg which was increased in SLE + AS mice, indicating that Bregs may regulate Th17/Treg cell homeostasis and cytokine release via IL-35 and TGF-ß production.

Retinoic Acid-Induced Regulation of Inflammatory Pathways Is a Potential Sepsis Treatment.

Sepsis is among the most dangerous known diseases, resulting from the dysregulation of the innate immune system in a process that is characterized largely by proinflammatory cytokines. It manifests as an excessive immune response to a pathogen and often leads to life-threatening complications such as shock and multiple-organ failure. Within the past several decades, much progress has been made to better understand the pathophysiology of sepsis and improve treatment. However, the average case-fatality rate for sepsis remains high. Current anti-inflammatory therapeutics for sepsis are not effective for use as first-line treatments. Focusing on all-trans-retinoic acid (RA), or activated vitamin A, as a novel anti-inflammatory agent, we have shown both in vitro and in vivo that RA decreases the production of proinflammatory cytokines. In vitro studies using mouse RAW 264.7 macrophages show that RA decreases tumor necrosis factor alpha (TNF-α) and interleukin-1ß (IL-1ß) and increases mitogen-activated protein kinase phosphatase 1 (MKP-1). RA treatment was also associated with the reduced phosphorylation of key inflammatory signaling proteins. Using a lipopolysaccharide and cecal slurry sepsis model, we found that RA significantly reduced mortality rates in mice, downregulated proinflammatory cytokine production, decreased neutrophil infiltration into lung tissue, and reduced the destructive lung histopathology typically seen in sepsis. We propose that RA may increase the function of native regulatory pathways and serve as a novel treatment for sepsis.

Phospholipase C-ε defines a PACAP-stimulated pathway for secretion in the chromaffin cell.

Adrenomedullary chromaffin cells respond to splanchnic (sympathetic) nerve stimulation by releasing stress hormones into the circulation. The signal for hormone secretion is encoded in the neurotransmitters - especially acetylcholine (ACh) and pituitary adenylate cyclase activating polypeptide (PACAP) - that are released into the splanchnic-chromaffin cell synapse. However, functional differences in the effects of ACh and PACAP on the chromaffin cell secretory response are not well defined. Here, selective agonists of PACAP receptors or nicotinic and muscarinic acetylcholine receptors were applied to chromaffin cells. The major differences in the effects of these agents were not on exocytosis, per se, but rather on the steps upstream of exocytosis. In almost every respect, the properties of individual fusion events triggered by PACAP and cholinergic agonists were similar. On the other hand, the properties of the Ca2+ transients evoked by PACAP differed in several ways from those evoked by muscarinic and nicotinic receptor stimulation. A defining feature of the PACAP-stimulated secretory pathway was its dependence on signaling through exchange protein directly activated by cAMP (Epac) and PLCε. However, the absence of PLCε did not disrupt Ca2+ transients evoked by cholinergic agonists. Accordingly, inhibition of Epac activity did not disrupt secretion triggered by acetylcholine or specific agonists of muscarinic and nicotinic receptors. Thus, PACAP and acetylcholine stimulate chromaffin cell secretion via separate and independent pathways. This feature of stimulus-secretion coupling may be important for sustaining hormone release from the adrenal medulla under conditions associated with the sympathetic stress response.

Synaptotagmin-7 facilitates acetylcholine release in splanchnic nerve-chromaffin cell synapses during nerve activity.

Disturbances that threaten homeostasis elicit activation of the sympathetic nervous system (SNS) and the adrenal medulla. The effectors discharge as a unit to drive global and immediate changes in whole-body physiology. Descending sympathetic information is conveyed to the adrenal medulla via preganglionic splanchnic fibers. These fibers pass into the gland and synapse onto chromaffin cells, which synthesize, store, and secrete catecholamines and vasoactive peptides. While the importance of the sympatho-adrenal branch of the autonomic nervous system has been appreciated for many decades, the mechanisms underlying transmission between presynaptic splanchnic neurons and postsynaptic chromaffin cells have remained obscure. In contrast to chromaffin cells, which have enjoyed sustained attention as a model system for exocytosis, even the Ca2+ sensors that are expressed within splanchnic terminals have not yet been identified. This study shows that a ubiquitous Ca2+-binding protein, synaptotagmin-7 (Syt7), is expressed within the fibers that innervate the adrenal medulla, and that its absence can alter synaptic transmission in the preganglionic terminals of chromaffin cells. The prevailing impact in synapses that lack Syt7 is a decrease in synaptic strength and neuronal short-term plasticity. Evoked excitatory postsynaptic currents (EPSCs) in Syt7 KO preganglionic terminals are smaller in amplitude than in wild-type synapses stimulated in an identical manner. Splanchnic inputs also display robust short-term presynaptic facilitation, which is compromised in the absence of Syt7. These data reveal, for the first time, a role for any synaptotagmin at the splanchnic-chromaffin cell synapse. They also suggest that Syt7 has actions at synaptic terminals that are conserved across central and peripheral branches of the nervous system.

Prevalence of Frailty and Associated Factors Among Hospitalized Older Adults: A Cross-Sectional Study.

This study aimed to estimate the prevalence of frailty and associated factors among hospitalized older adults. It consisted of 184 hospitalized older adults recruited between October 2019 to January 2020. We used the FRAIL scale, Geriatric Depression Scale-15, and the Pittsburg Sleep Quality Index to collect data. Descriptive statistics, independent group t-test, Chi-square (χ2) tests, and logistic regression were applied to statistical analysis. It found that the prevalence of frailty among hospitalized older adults was 39.7%. Depression (Mild: OR = 5.312, 95% CI [2.384-11.833]; Moderate: OR = 6.630, 95% CI [2.077-21.160]) and low activities of daily living (ADL) (Slight dependence: OR = 5.667, 95% CI [1.308-24.557]; Moderate dependence: OR = 15.188, 95% CI [3.342-69.016]; Severe dependence: OR = 5.872, 95% CI [2.645-13.038]) were independent predictors of frailty. Future studies on the interventions to reduce depression, improve ADL and delay the progression of frailty are encouraged. We should focus more on ADL, emotional and psychological state of hospitalized older adults to prevent frailty.

PACAP and acetylcholine cause distinct Ca2+ signals and secretory responses in chromaffin cells.

The adrenomedullary chromaffin cell transduces chemical messages into outputs that regulate end organ function throughout the periphery. At least two important neurotransmitters are released by innervating preganglionic neurons to stimulate exocytosis in the chromaffin cell-acetylcholine (ACh) and pituitary adenylate cyclase activating polypeptide (PACAP). Although PACAP is widely acknowledged as an important secretagogue in this system, the pathway coupling PACAP stimulation to chromaffin cell secretion is poorly understood. The goal of this study is to address this knowledge gap. Here, it is shown that PACAP activates a Gαs-coupled pathway that must signal through phospholipase C ε (PLCε) to drive Ca2+ entry and exocytosis. PACAP stimulation causes a complex pattern of Ca2+ signals in chromaffin cells, leading to a sustained secretory response that is kinetically distinct from the form stimulated by ACh. Exocytosis caused by PACAP is associated with slower release of peptide cargo than exocytosis stimulated by ACh. Importantly, only the secretory response to PACAP, not ACh, is eliminated in cells lacking PLCε expression. The data show that ACh and PACAP, acting through distinct signaling pathways, enable nuanced and variable secretory outputs from chromaffin cells.

Gut microbiota regulates chronic ethanol exposure-induced depressive-like behavior through hippocampal NLRP3-mediated neuroinflammation.

Chronic ethanol exposure (CEE), which can lead to neuroinflammation, is an increasing risk factor for depression disorder, but the underlying mechanism is not clear. Recent observations have revealed the associations among psychiatric disorders, ethanol exposure and alterations of the gut microbiota. Here, we found that CEE induced depressive-like behavior, which could be alleviated by probiotics and transferred from donor to recipient mice by fecal microbiota transplantation (FMT). Neuroinflammation and the activation of the NLRP3 inflammasome were also observed in recipient mice. The downregulation of NLRP3 in the hippocampus mitigated CEE-induced depressive-like behavior and neuroinflammation but had no significant effect on FMT recipient mice. Moreover, elevated serum inflammatory factors in recipient mice showed a significant mediation effect between the gut microbiota and depressive-like behavior. Together, our study findings indicate that the gut microbiota contributes to both hippocampal NLRP3-mediated neuroinflammation and depressive-like behavior induced by CEE, which may open avenues for potential interventions against CEE-associated psychiatric disorders.

Association Between Role Overload and Sleep Quality Among Chinese First-Line Nurses During the COVID-19 Outbreak: The Moderating Effect of Mindfulness.

BACKGROUND: Nurses working during the coronavirus disease 2019 (COVID-19) pandemic have experienced a high incidence of insomnia because of exposure to life-threatening occupational risks. Good sleep is essential for nurses to maintain their ability to care for patients with infectious diseases. PURPOSE: The aim of this study was to assess the influence of role overload on sleep quality and the moderating role of mindfulness. METHODS: A cross-sectional survey design was used in this study, which was conducted between March 20 and April 5, 2020. The survey was completed by 357 nurses who had relocated from Fujian Province to the epicenter of the outbreak in China to treat patients with COVID-19. Role overload, sleep quality, and mindfulness in these nurses were evaluated using the Role Overload Scale, Pittsburgh Sleep Quality Index, and Five-Facet Mindfulness Questionnaire, respectively. RESULTS: Hierarchical regression and other statistical methods were used to analyze the data. Role overload was shown to be positively related to poor sleep quality, and mindfulness was found to be effective in alleviating sleep disorders associated with role overload. CONCLUSIONS: The high risk of sleep disturbance among frontline nurses may be alleviated by reducing their perceived role overload. The identification of mindfulness as a moderating mechanism in the relationship between role overload and sleep quality provides new insights to improve sleep quality in nurses.

Chronic ethanol exposure induced anxiety-like behaviour by altering gut microbiota and GABA system.

Ethanol, also known as alcohol, is one of the most common drinks in the world. Chronic ethanol exposure has been reported to induce mental disorders. Ethanol also has a strong effect on the gut microbiota. The gut microbiota has been reported to affect the brain via multiple pathways, including changes in γ-aminobutyric acid (GABA) system, and cause a variety of mental disorders. The GABA system in the cortex is associated with anxiety. However, the role of gut microbiota played in ethanol exposure-induced changes in the GABA system and anxiety is still not clear. We established a 30-day ethanol exposure mouse model and investigated the effects of microbiota using the antibiotic minocycline. Minocycline alleviated ethanol-induced anxiety-like behaviour, dysbiosis of microbiota, intestinal barrier disruption, increased serum endotoxin and interleukin (IL)-6. Minocycline also attenuated ethanol-induced apoptosis and decreased expression of glutamate decarboxylases (GADs) and GABRA1 in the prefrontal cortex. Our results indicated that gut microbiota plays an important role in ethanol-induced anxiety-like behaviour by altering the function of GABA system. In addition, causal mediation analysis showed that endotoxin and IL-6 may mediate the connection between the gut microbiota and the expression of GABAA receptor in the prefrontal cortex.

Machine Learning-Based Overall Survival Prediction of Elderly Patients With Multiple Myeloma From Multicentre Real-Life Data.

Objective: To use machine learning methods to explore overall survival (OS)-related prognostic factors in elderly multiple myeloma (MM) patients. Methods: Data were cleaned and imputed using simple imputation methods. Two data resampling methods were implemented to facilitate model building and cross validation. Four algorithms including the cox proportional hazards model (CPH); DeepSurv; DeepHit; and the random survival forest (RSF) were applied to incorporate 30 parameters, such as baseline data, genetic abnormalities and treatment options, to construct a prognostic model for OS prediction in 338 elderly MM patients (>65 years old) from four hospitals in Beijing. The C-index and the integrated Brier score (IBwere used to evaluate model performances. Results: The 30 variables incorporated in the models comprised MM baseline data, induction treatment data and maintenance therapy data. The variable importance test showed that the OS predictions were largely affected by the maintenance schema variable. Visualizing the survival curves by maintenance schema, we realized that the immunomodulator group had the best survival rate. C-indexes of 0.769, 0.780, 0.785, 0.798 and IBS score of 0.142, 0.112, 0.108, 0.099 were obtained from the CPH model, DeepSurv, DeepHit, and the RSF model respectively. The RSF model yield best scores from the fivefold cross-validation, and the results showed that different data resampling methods did affect our model results. Conclusion: We established an OS model for elderly MM patients without genomic data based on 30 characteristics and treatment data by machine learning.

Internal structure design for improved shape fidelity and crispness of 3D printed pumpkin-based snacks after freeze-drying.

This study aimed to use the internal design of 3D food printing (3DFP) technology to obtain freeze-dried pumpkin with controllable crispness and higher shape fidelity. Two internal structural variables, namely filling pattern (honeycomb, rectilinear, grid, and triangular) and filling rate (25, 50, 75, and 100%), were studied to assess the impact on the shape fidelity and crispness characteristic of the product. As the filling rate decreased, the printing accuracy of the samples increased. Regardless of the filling patterns, the 75% filled samples exhibited the greatest deformation. The crispness of the samples was closely related to the filling pattern and filling rate. In the case of the high filling rate, the internal structure of the samples was dense. It was less likely to be broken under the action of force and the crispness was reduced. In addition, the internal structure of the sample influenced its physical properties, and the crispness customization of the product can be achieved by designing the porosity. Morphological differences between printed and cast samples suggested that 3DFP was beneficial for the processing and preparation of highly viscoelastic materials. The crispness of cast sample was obviously less than that of the 100% filled printed samples. The results opened an interesting perspective to create crisp foods with high shape fidelity that meet specific texture requirements and provide new sensory perceptions.