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Summary: Real-time confocal and super-resolution imaging reveals membrane dynamics of exo- and endocytosis, including hemi-fusion, fusion pore opening, expansion, constriction, closure (kiss-and-run), fused-vesicle shrinking (shrink fusion), and flat membrane transition to vesicles via intermediate Λ- and Ω-shape structures. Here, we describe a protocol for imaging these membrane dynamics, including primary culture of bovine adrenal chromaffin cells, fluorescent probe application, patch-clamp to deliver depolarization and evoke exo- and endocytosis, electron microscopy (EM), and real-time confocal and stimulated emission depletion (STED) microscopy.For complete details on the use and execution of this protocol, please refer to Zhao et al. (2016), Shin et al. (2018), and Shin et al. (2021). : Publisher's note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

The H9N2 avian influenza virus has become one of the dominant subtypes of avian influenza virus in poultry and has been significantly harmful to chickens in China, with great economic losses in terms of reduced egg production or high mortality by co-infection with other pathogens. A prediction of H9N2 status based on easily available production data with high accuracy would be important and essential to prevent and control H9N2 outbreaks in advance. This study developed a machine learning framework based on the XGBoost classification algorithm using 3 months’ laying rates and mortalities collected from three H9N2-infected laying hen houses with complete onset cycles. A framework was developed to automatically predict the H9N2 status of individual house for future 3 days (H9N2 status + 0, H9N2 status + 1, H9N2 status + 2) with five time frames (day + 0, day − 1, day − 2, day − 3, day − 4). It had been proven that a high accuracy rate > 90%, a recall rate > 90%, a precision rate of >80%, and an area under the curve of the receiver operator characteristic ≥ 0.85 could be achieved with the prediction models. Models with day + 0 and day − 1 were highly recommended to predict H9N2 status + 0 and H9N2 status + 1 for the direct or auxiliary monitoring of its occurrence and development. Such a framework could provide new insights into predicting H9N2 outbreaks, and other practical potential applications to assist in disease monitor were also considerable.

Abstract With the popularization of friction stir welding (FSW), 5083-H321 and 6061-T6 aluminum alloy materials are widely used during the FSW process. In this study, the fatigue life of friction stir welding with two materials, i.e., 5083-H321 and 6061-T6 aluminum alloy, are studied. Fatigue tests were carried out on the base metal of these two materials as well as on the butt joints and overlapping FSW samples. The principle of the equivalent structural stress method is used to analyze the FSW test data of these two materials. The fatigue resistances of these two materials were compared and a unified principal S–N curve equation was fitted. Two key parameters of the unified principal S–N curve obtained by fitting, C d is 4222.5, and h is 0.2693. A new method for an FSW fatigue life assessment was developed in this study and can be used to calculate the fatigue life of different welding forms with a single S–N curve. Two main fatigue tests of bending and tension were used to verify the unified principal S–N curve equation. The results show that the fatigue life calculated by the unified mean 50% master S–N curve parameters are the closest to the fatigue test results. The reliability, practicability, and generality of the master S–N curve fitting parameters were verified using the test data. The unified principal S–N curve acquired in this study can not only be used in aluminum alloy materials but can also be applied to other materials.

Abstract Working memory (WM) is a fundamental cognitive function that typically declines with age. Previous studies have shown that targeted WM training has the potential to improve WM performance in older adults. In the present study, we investigated whether a multi-domain cognitive training program that was not designed to specifically target WM could improve the behavioral performance and affect the neural activity during WM retrieval in healthy older adults. We assigned healthy older participants (70–78 years old) from a local community into a training group who completed a 3-month multi-domain cognitive training and a control group who only attended health education lectures during the same period. Behavioral and electroencephalography (EEG) data were recorded from participants while performing an untrained delayed match or non-match to category task and a control task at a pre-training baseline session and a post-training follow-up session. Behaviorally, we found that participants in the training group showed a trend toward greater WM performance gains than participants in the control group. Event-related potential (ERP) results suggest that the task-related modulation of P3 during WM retrieval was significantly enhanced at the follow-up session compared with the baseline session, and importantly, this enhancement of P3 modulation was only significant in the training group. Furthermore, no training-related effects were observed for the P2 or N2 component during WM retrieval. These results suggest that the multi-domain cognitive training program that was not designed to specifically target WM is a promising approach to improve WM performance in older adults, and that training-related gains in performance are likely mediated by an enhanced modulation of P3 which might reflect the process of WM updating.

Abstract Background Hypertension is the the primary cause of diastolic heart failure. Oxidative stress plays an important role in cardiac diastolic dysfunction caused by hypertension. The occurrence of oxidative stress is related to the level of nitric oxide (NO) in the body. Tetrahydrobiopterin (BH4) is an essential cofactor for NO synthesis. Nebivolol can reduce myocardial oxidative stress and increase NO activity. Therefore, we investigated the effects of monotherapy or combination therapy of different doses of BH4 and nebivolol on cardiac diastolic function in spontaneously hypertensive rats, and preliminarily expounded the related mechanisms. Methods Left ventricular function was evaluated by non-invasive echocardiographic assessment and invasive right carotid artery catheterization methods. ELISA was used to measure myocardial 3-nitrotyrosine content, NO production, and cyclic guanosine monophosphate (cGMP) concentration in the myocardium; quantitative real-time PCR (qRT-PCR) was used to determine endothelial nitric oxide synthase (eNOS), phospholamban and sarcoplasmic reticulum Ca2+-ATPase 2a (SERCA2a) mRNA expression levels; Western blot was used to detect the protein expression levels of eNOS and eNOS dimers in myocardial tissue, and immunohistochemical detection of cGMP expression in the myocardium was performed. Results Studies have shown that compared with those in the control group, NO generation and the expression level of myocardial eNOS mRNA, eNOS expression of dimers, phospholamban, SERCA2a and cGMP increased significantly after the combined intervention of BH4 and nebivolol, while the expression of 3-nitrotyrosine was significantly decreased. Conclusions The combined treatment group had a synergistic effect on reducing myocardial oxidative stress, increasing eNOS content, and increasing NO production, and had a more obvious protective effect on diastolic dysfunction through the nitric oxide/cyclic guanosine monophosphate (NO/cGMP) pathway.

ObjectiveAcupuncture has been shown to be effective in the treatment of chronic pain. However, their neural mechanism underlying the effective acupuncture response to chronic pain is still unclear. We investigated whether metabolic patterns in the pain matrix network might predict acupuncture therapy responses in patients with primary dysmenorrhea (PDM) using a machine-learning-based multivariate pattern analysis (MVPA) on positron emission tomography data (PET).MethodsForty-two patients with PDM were selected and randomized into two groups: real acupuncture and sham acupuncture (three menstrual cycles). Brain metabolic data from the three special brain networks (the sensorimotor network (SMN), default mode network (DMN), and salience network (SN)) were extracted at the individual level by using PETSurfer in fluorine-18 fluorodeoxyglucose positron emission tomography (18F-FDG-PET) data. MVPA analysis based on metabolic network features was employed to predict the pain relief after treatment in the pooled group and real acupuncture treatment, separately.ResultsPaired t-tests revealed significant alterations in pain intensity after real but not sham acupuncture treatment. Traditional mass-univariate correlations between brain metabolic and alterations in pain intensity were not significant. The MVPA results showed that the brain metabolic pattern in the DMN and SMN did predict the pain relief in the pooled group of patients with PDM (R2 = 0.25, p = 0.005). In addition, the metabolic pattern in the DMN could predict the pain relief after treatment in the real acupuncture treatment group (R2 = 0.40, p = 0.01).ConclusionThis study indicates that the individual-level metabolic patterns in DMN is associated with real acupuncture treatment response in chronic pain. The present findings advanced the knowledge of the brain mechanism of the acupuncture treatment in chronic pain.

Accurate wind resource assessments are necessary for cost effective offshore wind energy developments. The wind field offshore depends on the sea state. In coastal areas, where wind farms are usually built today, wind and waves are often not in full balance. In addition, wind farms modify their surrounding wind and turbulence field, especially downwind. These wind farm wakes, in turn, interact with the wave field, creating a complex dynamical system. To fully capture the dynamics in such a system in a realistic way, a coupled atmosphere-wave modelling system equipped with a wind farm parameterization should be applied. However, most conventional resource assessment relies on standalone atmosphere model simulations. We compare the wind-wave-wake climate predicted from a coupled modelling system, to one predicted from a standalone atmosphere model. Using a measurement-driven statistical-dynamical downscaling method, we show that about 180 simulation days are enough to represent the wind- and wave-climate, as well as the relation between those two, for the German Bight. We simulate these representative days with the atmosphere-wave coupled and the uncoupled modelling system. We perform simulations both without wind farms as well as parameterizing the existing wind farms as of July 2020. On a climatic average, wind resources derived from the coupled modelling system are reduced by 1% in 100 m over the sea compared to the uncoupled modelling system. In the area surrounding the wind farm the resources are further reduced. While the climatic reduction is relatively small, wind speed differences between the coupled and uncoupled modelling systems differ by more than ±20% on a 10-min time-scale. The turbulent kinetic energy derived from the coupled system is higher, which contributes to a more efficient wake dissipation on average and thus slightly smaller wake-affected areas in the coupled system. Neighbouring wind farms reduce wind resources of surrounding farms by up to 10%. The wind farm wakes reduce significant wave height by up to 3.5%. The study shows the potential of statistical-dynamical downscaling and coupled atmosphere-wave-wake modelling for offshore wind resource assessment and physical environmental impact studies.

Lighting conditions may affect the development of retinal degenerative diseases such as macular degeneration. In this study, to determine whether the lighting environment affects the progression of degeneration of retinal ganglion cells (RGCs), we examined glutamate/aspartate transporter (GLAST) heterozygous (GLAST+/-) mice, a mouse model of normal tension glaucoma. GLAST+/- mice were reared under a 12-h light-dark cycle (Light/Dark) or complete darkness (Dark/Dark) condition after birth. The total RGC number in the Dark/Dark group was significantly decreased compared with the Light/Dark group at 3 weeks old, while the number of osteopontin-positive αRGCs were similar in both groups. At 6 and 12 weeks old, the total RGC number were not significantly different in both conditions. In addition, the retinal function examined by multifocal electroretinogram were similar at 12 weeks old. These results suggest that lighting conditions may regulate the progression of RGC degeneration in some types of glaucoma.

Abstract Background Follicle selection in chickens refers to the process of selecting one follicle from a group of small yellow follicles (SY, 6–8 mm in diameter) for development into 12–15 mm hierarchical follicles (usually F6 follicles), which is an important process affecting laying performance in the poultry industry. Although transcriptomic analysis of chicken ovarian follicles has been reported, integrated analysis of chicken follicles for selection by using both transcriptomic and proteomic approaches is still rarely performed. In this study, we compared the proteomes and transcriptomes of SY and F6 follicles in laying hens and identified several genes involved in chicken follicle selection. Results Transcriptomic analysis revealed 855 differentially expressed genes (DEGs) between SY follicles and F6 follicles in laying hens, among which 202 were upregulated and 653 were downregulated. Proteomic analysis revealed 259 differentially expressed proteins (DEPs), including 175 upregulated and 84 downregulated proteins. Among the identified DEGs and DEPs, changes in the expression of seven genes, including VLDLR1, WIF1, NGFR, AMH, BMP15, GDF6 and MMP13, and nine proteins, including VLDLR, VTG1, VTG3, PSCA, APOB, APOV1, F10, ZP2 and ZP3L2, were validated. Further analysis indicated that the mRNA level of chicken VLDLR was higher in F6 follicles than in SY follicles and was also higher in granulosa cells (GCs) than in thecal cells (TCs), and it was stimulated by FSH in GCs. Conclusions By comparing the proteomes and transcriptomes of SY and F6 follicles in laying hens, we identified several differentially expressed proteins/genes that might play certain roles in chicken follicle selection. These data may contribute to the identification of functional genes and proteins involved in chicken follicle selection.

TAGAP gene variants are linked to human autoimmunity. Here the authors identify TAGAP as a Dectin-1 and EphB2-binding protein mediating antifungal innate immune signaling and cytokine production, and demonstrate TAGAP in non-T cells promotes Th17 response in mouse models of infection and autoimmunity.

Metabolomics has been used to explore the molecular mechanism and screen biomarkers. However, the critical metabolic signatures associated with benzene-induced hematotoxicity remain elusive. Here, we performed a plasma metabolomics study in 86 benzene-exposed workers and 76 healthy controls, followed by a validation analysis in mice, to investigate the dynamical change of the metabolic profile. We found that 8 fatty acids were significantly altered in both benzene-exposed worker and benzene-exposed animal models. These metabolites were significantly associated with S-phenylmercapturic acid and WBC, and they mediated the benzene-induced WBC decline. Furthermore, in vivo results confirm that fatty acid levels were dynamically altered, characterized by a decrease at 15 days and then sharp increases at 30 and 45 days. Following these identified fatty acids, the potential metabolic pathways were investigated. Fatty acids, as precursors for fatty acid oxidation, may disturb the balance of fatty acid biosynthesis and degradation. Our results reveal that fatty acid metabolism was strongly reprogrammed after benzene exposure. This abnormal change of fatty acids might be the key metabolic signature associated with benzene-induced hematotoxicity.

Pain is a multidimensional process, which can be modulated by emotions; however, the mechanisms underlying this modulation are unknown. We used pictures with different emotional valence (negative, positive, and neutral) as primes and applied electrical painful stimuli as targets to healthy participants. We assessed pain intensity and unpleasantness ratings and recorded electroencephalograms (EEGs). We found that pain unpleasantness and not pain intensity ratings were modulated by emotion, with increased ratings for negative and decreased ratings for positive pictures. We also found two consecutive gamma band oscillations (GBOs) related to pain processing from time frequency analyses of the EEG signals. The early GBO had a cortical distribution contralateral to the painful stimulus and its amplitude was positively correlated with intensity and unpleasantness ratings, but not with prime valence. The late GBO had a centroparietal distribution and its amplitude was larger for negative compared to neutral and positive pictures. The emotional modulation effect (negative vs. positive) of the late GBO amplitude was positively correlated with pain unpleasantness. The early GBO might reflect the overall pain perception, possibly involving the thalamocortical circuit, while the late GBO might be related to the affective dimension of pain and top-down-related processes.

Insect identification is the basis of insect research and disaster control and is of great importance for the design of pest control strategies and the protection of beneficial insects. Due to human subjective limitations and the small size and uneven distribution of pests, traditional methods of distinguishing and counting pest types based on experience cannot quickly and accurately detect and identify pests. Therefore, this paper proposes an object detection algorithm based on the improved Mask R-CNN model, aiming to improve the accuracy and efficiency in pest identification and counting. The algorithm improves the FPN structure in the feature extraction network and increases the weight coefficient when fusing feature layers of different scales. Based on the task of target detection and recognition, weight coefficient is adjusted to a proper parameter so that the semantic information and positioning information can be made full use to achieve more accurate recognition and positioning. The results of the experimental analysis of 1000 sample images show that the improved Mask R-CNN model has a recognition and detection accuracy of 99.4%, which is 2.7% higher than that of the unimproved Mask R-CNN model. The main contribution of this method is to improve the detection speed, and at the same time, the recognition accuracy has been significantly improved. This algorithm provides technical support for pest detection in the agricultural field and makes a contribution to the intellectualization of agricultural management.

Deception emerges in early childhood and prevails in adults. Activation patterns in previous adults’ task-state functional magnetic resonance imaging (fMRI), though sensitive to state honesty on a specific decision, are less reliable reflecting trait honesty. Besides of state honesty, most previous neuroimaging studies about dishonesty suffer the generalization problem due to the major focus on adults with children unexplored. To investigate honesty associated functional brain networks variations, 98 healthy adults (Age: 18–28 y.o.; 49 males and 49 females) were invited to participate in a resting-state functional magnetic resonance imaging (rfMRI) study (Study 1). We investigated how functional connections between the caudate and the medial prefrontal cortex (mPFC) change among adults who differ in self-reported trait honesty. Results showed that adults with higher trait honesty have increased functional connectivity from the caudate to the mPFC, which is identified as an honesty-related hub region in global brain connectivity analysis and connects more tightly to a wide range of brain regions including the amygdala. Study 2 compared functional connectivity between children with high vs. low lying frequencies (Age: 6–16 y.o.; 61 males and 39 females) based on a publicly accessible database of rfMRI. Consistent with findings in adults, increased functional connectivity from the caudate to the mPFC was found in less frequently lying children. Despite different honesty indicators of self-reported honesty trait in adults and parent-reported lying patterns in children, consistent findings have been noted in the two samples with regards to functional connectivity variations between reward-related and self-related brain regions. These findings suggest functional connectivity alterations between the caudate and the mPFC contribute to honesty variations in both adults and children.

Backgrounds: Intra-abdominal placement of the Central Venous Catheter {QJQ was conducted to manage the ascites-related symptoms of non-ovarian cancer patients. The aim of this study is to document the efficacy of symptom relief and conduct survival analysis of non-ovarian cancer patients with malignant ascites who received paracentesis and indwelling catheter drainage. Methods: Seventy eight patients received paracentesis and drainage. All patients who met the inclusion criteria were included in this study. The overall survival (OS) was defined as the interval between initial diagnosis and death. Since-paracentesis survival (SP-Survival) was defined as the interval between initial paracentesis and death. Results: Hepatic cancer was the most frequent original cancer in this study. Peritoneal catheters remained in situ for a median of 13 days. No immediate complications, such as perforation of a viscus or excessive bleeding, were encountered during placement. All ascites-related symptoms improved after drainage compared with the baseline. There was a statistically significant improvement in the mean score for abdominal swelling (p< 0.001), anorexia (p = 0.023) and constipation (p = 0.045). Cancer type was shown to be an independent prognostic factor for overall survival length (p = 0.001). Serum albumin was an independent prognostic factor for SP-survival (p = 0.02). Conclusions: Paracentesis and indwelling catheter drainage through CVC set is a useful method for improving painful symptom. Further research is needed to validate the findings. [ABSTRACT FROM AUTHOR]

Background: Long term low-dose benzene exposure leads to the inhibition of haematopoiesis. However, the underlying mechanisms remained poorly defined, especially mediated by early effector molecules. Methods: Here, we first found in mRNA microarray that pyroptotic classic genes (Casp1, 4, 5, and IL1β) were up-regulated and represented dose-dependent differential expression in controls, low-dose benzene-exposed and chronic benzene-poisoned workers, and the expression of Casp1 and IL1β were confirmed in low-dose benzene-exposed workers and was accompanied with elevated potent proinflammatory IL1β. In vitro studies showed that benzene metabolites induced AHH-1 cell pyroptosis through activating Aim2/Casp1 pathway with the increased expression of GSDMD. Meanwhile, TET2 overexpression was elevated in vivo and in vitro and it was positively correlated with IL1β. Further, we verified that pyroptosis caused by 1,4-BQ could be ameliorated in vitro by RNAi or pretreatment with Dimethyloxalylglycine (DMOG), the inhibitor of TET2. Findings: Exposure to benzene can trigger pyroptosis via TET2 directly regulating the Aim2/Casp1 signaling pathway to cause haematotoxicity. Interpretation: Benzene metabolites induced pyroptotic cell death through activation of the Aim2/Casp1 pathway which can be regulated by Tet2 overexpression. Tet2 may be a potential risk factor and is implicated in the development of benzene-related diseases. Fund: National Natural Science Foundation of China; the Support Project of High–level Teachers in Beijing Municipal Universities in the Period of 13th Five–year Plan; Beijing Natural Science Foundation Program and Scientific Research Key Program of Beijing Municipal Commission of Education. Keywords: Benzene, TET2, Pyroptosis, Inflammatory, Hematotoxicity

This paper presents a simple formal theoretical model to explain why citizens in authoritarian regimes trust the illiberal official media more than the commercial media. Media trust is defined as changes in the citizen’s belief based on good or bad news from the media. Using this definition, the model evaluates the independent and interaction effect of media bias, censorship, media quality, the citizen’s prior belief of the situation, and the citizen’s ideology on media trust. The findings reconcile some controversies in the literature, and, more importantly, reveal new and subtle explanations the literature did not identify and probably needs to pay attention to.

Primary dysmenorrhea (PDM) is a common gynecological disease characterized by lower abdominal pain. Acupuncture is considered a good alternative therapy for PDM. However, the central mechanism of the analgesic effect of acupuncture is largely unknown. In this study, eligible patients were randomized into the real and sham acupuncture groups using a computer-generated, permuted block randomization method. The study cohort comprised 34 patients: 19 in the real acupuncture group and 15 in the sham acupuncture group. The clinical characteristics of the patients during their menstrual period were collected, and imaging scans were performed during the first 3 days of the patients’ menstrual period. We analyzed task and resting functional magnetic resonance imaging (fMRI) data to investigate the potential central mechanism of the immediate effect of acupuncture intervention on the intensity of PDM pain. The task fMRI study found that the rostral anterior cingulate cortex (rACC) and right supplemental motor area were activated during real acupuncture. Using the resting-state functional connectivity (FC) method, we found a post- versus pre-treatment change in the FC of the rACC and left precentral gyrus in the comparison of real acupuncture versus sham acupuncture. In addition, the FC of the rACC–left precentral gyrus at baseline was negatively correlated with short-term analgesia, while the change in the FC of the rACC–left precentral gyrus was positively correlated with short-term analgesia after acupuncture treatment. These findings support the importance of rACC–left precentral gyrus resting-state FC in the modulation of the intensity of PDM pain through acupuncture, which may shed light on the central mechanism of acupuncture in the treatment of PDM.

Background: There are a number of studies dedicated to characteristics of sedation, but these studies are mostly bound to western country practices. The aim of this study is to describe the characteristics of patients who suffered from cancer and who had been sedated until their death in Shanghai, China. Methods: Retrospective medical data of 244 terminally ill cancer patients including 82 sedated patients were collected. Data collected included demographic characteristics, disease-related characteristics and details of the sedation. Results: In sedated cases, patients and/or caregivers gave the consent to start palliative sedation due to unmanageable symptoms. On average, sedation was performed 24.65(±1.78)hours before death. Agitated delirium and dyspnea were the most frequent indications for palliative sedation. There was no significant difference in survival time from admission till death between sedated and non-sedated patients (p > 0.05). Conclusions: Palliative sedation is effective for reducing terminally ill cancer patients' suffering without hastening death. Prospective research is needed to determine the optimal conditions for Chinese patients including indications, decision making process, informed consent, cultural and ethical issues, type of sedation and drugs. [ABSTRACT FROM AUTHOR]

Background: Low benzene exposure leads to hematotoxicity, but we still lack sensitive early monitoring and early warning markers. Benzene is associated with inflammation, which is mainly mediated by cytokines network. However, until now few studies have conducted high-throughput detection of multi-cytokines to get a global view of cytokine changes and screen for markers of benzene-induced toxicity. We hypothesized that cytokine profiles mediate benzene-induced hematotoxicity. Methods: 228 subjects consisting of 114 low benzene exposed workers and 114 healthy controls were recruited at Research Center of Occupational Medicine, Peking University Third Hospital, Beijing. The serum concentrations of 27 cytokines were detected by cytokinomics array, urinary benzene series metabolites were measured by UPLC-MS/MS, and peripheral blood cell counts were observed by basic blood test. Results: Among 27 cytokines, IL-9 and MIP1-α were significantly lower, but IL-4, IL-10, IL-15, MCP-1, TNF-α and VEGF were significantly higher in benzene exposure group than controls. Urinary benzene metabolite S-phenylmercapturic acid (S-PMA) was significantly higher in benzene exposure group and had a negative linear relationship with WBC count. S-PMA was only significantly associated with IL-9, meanwhile IL-9, IL-15 and VEGF had a positive linear relationship with WBC count. The bootstrapping mediation models showed that the effect of S-PMA on WBC count was partially explained by IL-9 for 10.11%. Conclusion: This study suggests that exposure to benzene was associated with alternation of blood cell count and cytokine profiles in workers exposed to low levels of benzene, especially decreases of WBC count and IL-9. We also found IL-9 partially mediated the effect of low benzene exposure on WBC count, which may be a potential and promising early monitoring and early warning marker of benzene hematotoxicity.

Grouting reinforcement is one of the most effective methods to enhance the stability of the goaf, and its scheme selection, hole location, and parameter determination directly affect the success or failure of goaf treatment. On the basis of discussion of the deformation mechanism and evolution law of the longwall goaf, this article comprehensively analyzed the grouting reinforcement mechanism of the goaf combined with filling theory, permeability theory, and fracturing theory and studied the physical and chemical reaction principles of two commonly used filling materials, cement-fly ash slurry and cement-clay slurry. Three grouting models have been established: whole grouting, local grouting in fracture zone, and strip grouting, and then simulated the grouting effect of the two more common methods of whole grouting and strip grouting by numerical simulation software (FLAC3D, tecplot, Surfer). Simulation analysis is carried out from the perspectives of settlement deformation, horizontal movement, vertical stress, and horizontal stress. Finally, the feasibility of grouting treatment in the goaf is verified by engineering example. The results show that a reasonable grouting scheme can effectively reduce the residual deformation of the goaf and its overlying rock and improve the poor stress state, so as to achieve the purpose of effective grouting.

Anti-programmed cell death-1 (PD-1)/PD-ligand-1 (PD-L1) treatments are effective in a fraction of patients with advanced malignancies. However, the majority of patients do not respond to it. Resistance to cancer immunotherapy can be mediated by additional immune checkpoints. We hypothesized that co-targeting of PD-L1 and lymphocyte-activation gene 3 (LAG-3) could provide an alternative therapeutic approach. Here, we developed IBI323, a dual blockade bispecific antibody targeting PD-L1 and LAG-3. We assessed the binding affinity, blocking activity, cell bridging effect, and immunomodulation function of IBI323 using in vitro assays. We also evaluated, in two humanized mouse models, anti-tumor effects and antitumor T cell immunity induced by IBI323. IBI323 bound to PD-L1 and LAG-3 with similar potency as its parental antibodies and blocked the interaction of PD-1/PD-L1, CD80/PD-L1, and LAG-3/MHC-II. Moreover, IBI323 mediated the bridging of PD-L1+ cells and LAG-3+ cells and demonstrated superior immune stimulatory activity compared to each parent antibody in mixed leukocyte reaction. In PD-L1/LAG-3 double knock-in mice bearing human PD-L1 knock-in MC38 tumors, IBI323 showed stronger anti-tumor activity compared to each parental antibody. The better antitumor response correlated with increased tumor-specific CD8+ and CD4+ T cells. IBI323 also induced stronger anti-tumor effect against established A375 tumors compared with combination in mice reconstituted with human immune cells. Collectively, these data demonstrated that IBI323 preserved the blockade activities of parental antibodies while processing a novel cell bridging function. Based on the encouraging preclinical results, IBI323 has significant value in further clinical development.

The pain and fear caused by direct local injection of anesthetic or the poor experience with surface anesthetic cream increase the difficulty of clinical treatment for oral diseases. To address this problem, a hyaluronic acid microneedle patch (Li-HAMNs) that consists of fast-dissolving lidocaine hydrochloride (LDC)-loaded tips and a wet-adhesive backing layer made of polyvinyl alcohol (PVA)/carboxymethylcellulose sodium (CMC-Na) was fabricated to explore its potential use in dental topical anesthesia. Li-HAMNs could puncture the stratum corneum with an insertion depth of about 279 μm in the isolated porcine oral mucosal. The fast-dissolving tips could release LDC to improve the patients’ convenience and compliance. Importantly, the backing layer, which has good adhesion ability and water-absorbing properties, could surmount the contraction and extension of oral masticatory muscles and the saliva scour. In the tail flick test, the topical anesthesia efficacy of the Li-HAMNs group was much better than clinical lidocaine cream (EMLA cream, LDC, 1.2 mg) in spite of a relatively lower LDC dose with Li-HAMNs (LDC, 0.5 mg). It is believed that the proposed adhesive microneedle patch could enhance transmucosal delivery of anesthetics and thus open a new chapter in the painless treatment of oral diseases.

Primary dysmenorrhea (PDM) is a common complaint in women throughout the menstrual years. Acupuncture has been shown to be effective in dysmenorrhea; however, there are large interindividual differences in patients’ responses to acupuncture treatment. Fifty-four patients with PDM were recruited and randomized into real or sham acupuncture treatment groups (over the course of three menstrual cycles). Pain-related functional connectivity (FC) matrices were constructed at baseline and post-treatment period. The different neural mechanisms altered by real and sham acupuncture were detected with multivariate analysis of variance. Multivariate pattern analysis (MVPA) based on a machine learning approach was used to explore whether the different FC patterns predicted the acupuncture treatment response in the PDM patients. The results showed that real but not sham acupuncture significantly relieved pain severity in PDM patients. Real and sham acupuncture displayed differences in FC alterations between the descending pain modulatory system (DPMS) and sensorimotor network (SMN), the salience network (SN) and SMN, and the SN and default mode network (DMN). Furthermore, MVPA found that these FC patterns at baseline could predict the acupuncture treatment response in PDM patients. The present study verified differentially altered brain mechanisms underlying real and sham acupuncture in PDM patients and supported the use of neuroimaging biomarkers for individual-based precise acupuncture treatment in patients with PDM.

PurposeIt has been demonstrated that, for various types of cancer, the pretreatment albumin/alkaline phosphatase ratio (AAPR) was a prognostic factor. Therefore, in order to determine AAPR's prognostic effect on cancer, the meta-analysis was hereby performed.Patients and methodsThe relevant studies conducted before November 10, 2019, were comprehensively searched in Web of Science, PubMed, and Embase. HRs(hazard ratios) with related 95%CIs(confidence intervals) were adopted to estimate AAPR's prognostic impact on overall survival (OS) & disease-free survival (DFS).ResultsOur meta-analysis involved thirteen cohort studies, which included 5,204 cases of 8 types. The results of this meta-analysis indicated that higher AAPR was corrected with better OS (pooled HR = 0.52; 95%CI = 0.47-0.58; PConclusionHigher AAPR was related to better prognosis of cancer, and in cancer therapy, AAPR could be taken as a promising marker of prognosis. It might help physicians to select the most appropriate treatments by evaluating the current status of patients with cancer. Future multicenter prospective clinical trials were required to verify its applications.

Because of the arid climate and fragile ecological environment in Xinjiang, China, land reclamation should be carried out after mining. The core of land reclamation is the water content of the surface covering soil. In this paper, the law of water distribution in reclamation reconstructed soil of nonmetal mines in Xinjiang was studied. In order to obtain the law of water distribution in reconstructed soil, we set up an observation system of the neutron probe and tensiometer. The neutron probe was used to monitor the soil water content. The tensiometers were used to obtain the matrix potential of soil for verifying the water distribution in reconstructed soil. Volumetric water content and matrix potential of reconstructed soil during 1-year period of management and irrigation were obtained by long-term monitoring. After one year’s field in situ test, 2424 sets of neutron probe data and 1368 sets of tensiometer data were obtained. By studying the above parameters, we summarized the law of water distribution in reconstructed soil of variable thickness and degree of compaction with nonmetallic waste rock filling. The results showed that covering soil was helpful to retain water content. Whether the soil was compacted or uncompacted, the soil water content at the depth of 10 cm was less than that at other depth of reconstructed soil because it was greatly affected by meteorological factors. The water content of reconstructed soil at 30 cm depth was greater than that at other depths. Under the influence of factors such as the thickness and compaction of the soil, the response time of soil water content and matrix potential to each irrigation infiltration was different. According to the characteristics of reclamation-vegetation such as alfalfa growth in Xinjiang, the thickness of surface reconstructed soil should be not less than 50 cm. Over time, soil that was compacted once was better for the vegetation. The research results could provide a reference for the land reclamation of nonmetallic mines in Xinjiang, China.

Summary: For decades, two fusion modes were thought to control hormone and transmitter release essential to life; one facilitates release via fusion pore dilation and flattening (full collapse), and the other limits release by closing a narrow fusion pore (kiss-and-run). Using super-resolution stimulated emission depletion (STED) microscopy to visualize fusion modes of dense-core vesicles in neuroendocrine cells, we find that facilitation of release is mediated not by full collapse but by shrink fusion, in which the Ω-profile generated by vesicle fusion shrinks but maintains a large non-dilating pore. We discover that the physiological osmotic pressure of a cell squeezes, but does not dilate, the Ω-profile, which explains why shrink fusion prevails over full collapse. Instead of kiss-and-run, enlarge fusion, in which Ω-profiles grow while maintaining a narrow pore, slows down release. Shrink and enlarge fusion may thus account for diverse hormone and transmitter release kinetics observed in secretory cells, previously interpreted within the full-collapse/kiss-and-run framework. : Shin et al. discover two fusion modes; one involves fused vesicle shrinking that employs a large pore to facilitate content release, and the other involves vesicle enlargement with a small pore that slows down release. Shrinking is energetically preferred over the generally assumed full-collapse fusion, because osmotic pressure squeezes fused vesicles. Keywords: exocytosis, transmitter release, hormone release, osmotic pressure, membrane dynamics, membrane fusion, vesicle shrinking, vesicle enlargement, bovine chromaffin cells, mathematical modeling, mathematical modeling

In this paper, we propose a new type of non-recursive Mastrovito multiplier for GF(2m) using an n-term Karatsuba algorithm (KA), where GF(2m) is defined by an irreducible trinomial, xm+xk +1, m = nk. We show that such a type of trinomial combined with the n-term KA can fully exploit the spatial correlation of entries in related Mastrovito product matrices and lead to a low-complexity architecture. The optimal parameter n is further studied. As the main contribution of this paper, the lower bound of the space complexity of our proposal is about O(m2/2) + m3/2). Meanwhile, the time complexity matches the best Karatsuba multiplier known to date. To the best of our knowledge, it is the first time that Karatsuba-based multiplier has reached such a space complexity bound while maintaining a relatively low time delay.

In this manuscript, the influence of organosolv beech wood lignin (LOB) on its oxidative conversion to high added-value phenolic aldehydes is discussed. Environmental friendly and low-cost H2O2 was used as the oxygen atom donor. The catalyst was prepared by immobilizing Lanthanum compounds onto the periodic mesoporous channels of siliceous SBA-15. The activity of the La/SBA-15 was investigated towards oxidation of LOB in the presence of hydrogen peroxide as oxidant with microwave irradiation. Considering the characteristics of LOB, an unexpected low syringaldehyde concentration at 10min of reaction time (1.47 g/L, corresponding to 15.66% yield) was obtained; the other major product was vanillin at 25min (0.78 g/L, i.e., 9.94% yield). The high reactivity of syringyl nuclei may be pointed out as the main reason for the faster production and degradation of syringaldehyde in oxidation. Other low molecular weight phenolic products were found: vanillic acid, syringic acid and minor quantities of aceto-derivatives. The profile of products concentration with the reaction time of catalytic oxidation with microwave irradiation are shown and discussed with reference to the investigated lignin features. The mechanism of the microwave catalytic oxidation for LOB under alkaline conditions was proposed. [ABSTRACT FROM AUTHOR]

Strigolactones (SLs) and their derivatives are plant hormones that have recently been identified as regulators of primary lateral root (LR) development. However, whether SLs mediate secondary LR production in rice (Oryza sativa L.), and how SLs and auxin interact in this process, remain unclear. In this study, the SL-deficient (dwarf10) and SL-insensitive (dwarf3) rice mutants and lines overexpressing OsPIN2 (OE) were used to investigate secondary LR development. The effects of exogenous GR24 (a synthetic SL analogue), 1-naphthylacetic acid (NAA; an exogenous auxin), 1-naphthylphthalamic acid (NPA; a polar auxin transport inhibitor), and abamine (a synthetic SL inhibitor) on rice secondary LR development were investigated. Rice d mutants with impaired SL biosynthesis and signaling exhibited increased secondary LR production compared with wild-type (WT) plants. Application of GR24 decreased the numbers of secondary LRs in dwarf10 (d10) plants but not in dwarf3 (d3), plants. These results indicate that SLs negatively regulate rice secondary LR production. Higher expression of DR5::GUS and more secondary LR primordia were found in the d mutants than in the WT plants. Exogenous NAA application increased expression of DR5::GUS in the WT, but had no effect on secondary LR formation. No secondary LRs were recorded in the OE lines, although DR5::GUS levels were higher than in the WT plants. However, on application of NPA, the numbers of secondary LRs were reduced in d10 and d3 mutants. Application of NAA increased the number of secondary LRs in the d mutants. GR24 eliminated the effect of NAA on secondary LR development in the d10, but not in the d3, mutants. These results demonstrate the importance of auxin in secondary LR formation, and that this process is inhibited by SLs via the D3 response pathway, but the interaction between auxin and SLs is complex.

In hens, follicle selection is an important process affecting egg laying traits. This study investigated the role of parathyroid hormone-like hormone (PTHLH) in chicken follicle selection, its transcriptional regulation and genetic effects on egg laying traits. PTHLH and its receptor PTH1R were mainly expressed in follicles of 6–8 mm in diameter, exhibits differential expression pattern in the theca and granulosa cells of pre- and hierarchal follicles. PTHLH stimulates the proliferation of follicular granulosa and theca cells, the expression of StAR and CYP11A1 mRNA and the production of progesterone (P4) in pre-hierarchal follicles. Treatment with FSH increased PTHLH mRNA expression in pre-hierarchal follicular theca cells and hierarchal follicular granulosa cells. Two critical regions regulating chicken PTHLH transcription were revealed, each of which harbored a SNP: C>T (chr1: 72530014) for AP-1 and a SNP: A>G (chr1: 72531676). Hens with diplotype AC/GT were younger at first laying and laid more eggs at 32 weeks. The haplotype (G-1827T-165) with double mutations had the greatest promoter activity of chicken PTHLH transcription. Collectively, PTHLH plays an important role in chicken follicle selection by stimulating cell proliferation and steroidogenesis. Polymorphisms in chicken PTHLH promoter region are associated with egg laying traits by affecting the binding of transcription factor AP-1.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Amino acid L-histidine was introduced to the preparation of zeolitic imidazolate framework ZIF-L with controllable morphologies. Thick leaf-like particles (23µm in length) with a thickness of about 1 µm were prepared, and their crystallinity decreased when less L-histidine was used. The decreased amount of water in the precursor solution resulted in a morphology change from a leaf-like structure to irregular microsized particles. The regulation of ZIF crystal morphology will lead to more versatile applications of ZIFs as functional materials in separation and adsorption. [ABSTRACT FROM AUTHOR]

Glaucoma is a neurodegenerative disease of the eye, which involves degeneration of retinal ganglion cells (RGCs): the output neurons of the retina to the brain, which with their axons comprise the optic nerve. Recent studies have shown the possible involvement of oxidative stress in the pathogenesis of glaucoma, especially in the subtype of normal tension glaucoma. Basic experiments utilizing rodent and primate models of glaucoma revealed that antioxidants protect RGCs under various pathological conditions including glutamate neurotoxicity and optic nerve injury. These results suggested that existing drugs and food factors may be useful for prevention and hence therapy of glaucoma. In this review, we highlight some therapeutic candidates, particularly those with antioxidant properties, and discuss the therapeutic potential of RGC protection by modulating gene expressions that prevent and ameliorate glaucoma.

The adsorption mechanisms of mercury ion (Hg2+) by different fractions of biochar were studied, providing a theoretical basis and practical value for the use of biochar to remediate mercury contamination in water. Biochar (RC) was prepared using corn straw as the raw material. It was then fractionated, resulting in inorganic carbon (IC), organic carbon (OC), hydroxyl-blocked carbon (BHC), and carboxyl-blocked carbon (BCC). Before and after Hg2+ adsorption, the biochar fractions were characterized by several techniques, such as energy-dispersive X-ray spectroscopy (EDS), Fourier-transform infrared spectroscopy (FTIR), and X-ray photoelectron spectroscopy (XPS). Obtained results indicate that the reaction mechanisms of RC for Hg2+ removal mainly include electrostatic adsorption, ion exchange, reduction, precipitation, and complexation. The equilibrium adsorption capacity of RC for Hg2+ is 75.56 mg/g, and the adsorption contribution rates of IC and OC are approximately 22.4% and 77.6%, respectively. Despite the lower rate, IC shows the largest adsorption capacity, of 92.63 mg/g. This is attributed to all the mechanisms involved in Hg2+ adsorption by IC, with ion exchange being the main reaction mechanism (accounting for 39.8%). The main adsorption mechanism of OC is the complexation of carboxyl and hydroxyl groups with Hg2+, accounting for 71.6% of the total OC contribution. BHC and BCC adsorb mercury mainly via the reduction–adsorption mechanism, accounting for 54.6% and 54.5%, respectively. Among all the adsorption mechanisms, the complexation reaction of carboxyl and hydroxyl groups with Hg2+ is the dominant effect.

The dumps in the open-pit mining area in the eastern grassland are prone to landslides due to the fragile ecological environment, so it is inevitable to reshape the dump slopes. In order to explore a more scientific method for slope shaping of open-pit mine dump, slope stability analysis were used to compare effect of three types of slope-type (wave-shaped, slope-shaped and step-shaped slope shaping method)in outside dumping site of Baori Hiller open-pit mine. The results show that the slope stability is negatively correlated with the slope angle, and the stability of different shaping slopes is realized as wave-shaped slope (F=2.711)> Slope-shaped slope(F=2.513)>Step-shaped slope(F=1.047), in which the wave type and slope type are all within the safe range, but the step type slope is unstable; in consideration of cost, stability and erosion resistance, it is better to set the slope angle of the dump to 15°.The wave-shaped shaping method of the natural dumping of the excavation field outside the Baori Hiller open-pit mine has the best effect and is worth promoting.

The Jining Gray goat is famous for its sexual precocity; however, the exact regulatory mechanism is still unknown. The hypothalamus is the key centrum in the process of animal reproduction, especially in signal transduction, and the initiation of puberty. The identification of potential genes and pathways in the hypothalamus of Jining Gray goat is critical to understanding the regulatory mechanism of sexual precocity in these goats. In this study, mRNA transcriptome analysis of the hypothalamus of juvenile and pubertal goats revealed eight genes (NTS, ADORA1, CRH, UCN3, E2F2, PDGFRB, GNRH1, and CACNA1C) and three pathways [neuroactive ligand-receptor interaction; gonadotropin-releasing hormone (GnRH) signal; melanoma] that are involved in this regulation. Subsequent methylation analysis on differentially methylated region (DMR) genes revealed the potential regulation network that influences pubertal onset. Correlation analysis verified the methylation level of some DMR genes correlates negatively with expression level. Integrated analysis between transcriptomes and methylomes identified 80 candidate genes involved in GnRH and neuroactive ligand signal pathways, of which CACNA1C and CRH were differentially expressed genes (DEGs) influenced by methylation level. The GnRH gene was the only DEG not affected by its methylation level. In summary, in this study, we identified eight genes and three pathways that are related to pubertal onset in Jining Gray goats, and the expression of CACNA1C and CRH genes of the GnRH and neuroactive ligand signal pathways were influenced by DNA methylation, while that of the GnRH gene was not affected.

Polarized sorting of newly synthesized proteins to the somatodendritic and axonal domains of neurons occurs by selective incorporation into distinct populations of vesicular transport carriers. An unresolved issue is how the vesicles themselves are sorted to their corresponding neuronal domains. Previous studies concluded that the axon initial segment (AIS) is an actin-based filter that selectively prevents passage of somatodendritic vesicles into the axon. We find, however, that most somatodendritic vesicles fail to enter the axon at a more proximal region in the axon hillock, herein referred to as the pre-axonal exclusion zone (PAEZ). Forced coupling of a somatodendritic cargo protein to an axonally directed kinesin is sufficient to drive transport of whole somatodendritic vesicles through the PAEZ toward the distal axon. Based on these findings, we propose that polarized sorting of transport vesicles occurs at the PAEZ and depends on the ability of the vesicles to acquire an appropriately directed microtubule motor.

In high wind speed conditions, sea spray generated by intensely breaking waves greatly influences the wind stress and heat fluxes. Measurements indicate that the drag coefficient decreases at high wind speeds. The sea spray generation function (SSGF), an important term of wind stress parameterisation at high wind speeds, is usually treated as a function of wind speed/friction velocity. In this study, we introduce a wave-state-dependent SSGF and wave-age-dependent Charnock number into a high wind speed–wind stress parameterisation. The newly proposed wind stress parameterisation and sea spray heat flux parameterisation were applied to an atmosphere–wave coupled model to study the mid-latitude storm development of six storm cases. Compared with measurements from the FINO1 platform in the North Sea, the new wind stress parameterisation can reduce wind speed simulation errors in the high wind speed range. Considering only sea spray impact on wind stress (and not on heat fluxes) will intensify the storms (in terms of minimum sea level pressure and maximum wind speed), but has little effect on the storm tracks. Considering the impact of sea spray on heat fluxes only (not on wind stress) can improve the model performance regarding air temperature, but it has little effect on the storm intensity and storm track performance. If the impact of sea spray on both the wind stress and heat fluxes is taken into account, the model performs best in all experiments for minimum sea level pressure, maximum wind speed and air temperature.

Ovarian follicle selection is an important process impacting the laying performance and fecundity of hens, and is regulated by follicle-stimulating hormone (FSH) through binding to its receptor [follicle-stimulating hormone receptor (FSHR)]. In laying hens, the small yellow follicle (6–8 mm in diameter) with the highest expression of FSHR will be recruited into the preovulatory hierarchy during ovarian follicle development. The study of molecular mechanism of chicken follicle selection is helpful for the identification of genes underlying egg-laying traits in chicken and other poultry species. Herein, the transcriptomes of chicken small yellow follicles differing in the mRNA expression of FSHR were compared, and a total of 17,993 genes were identified in 3 pairs of small yellow follicles. The Wnt signaling pathway was significantly enriched in the follicles with the greatest fold change in FSHR expression. In this pathway, the expression level of Wnt4 mRNA was significantly upregulated with a log2(fold change) of 2.12. We further investigated the expression, function, and regulation of Wnt4 during chicken follicle selection and found that Wnt4 mRNA reached its peak in small yellow follicles; Wnt4 stimulated the proliferation of follicular granulosa cells (GCs), increased the expression of StAR and CYP11A1 mRNA in prehierarchical and hierarchical follicles, increased the expression of FSHR mRNA, and decreased the expression of anti-Müllerian hormone and OCLN mRNA. Treatment with FSH significantly increased Wnt4 expression in GCs. Moreover, Wnt4 facilitated the effects of FSH on the production of progesterone (P4) and the mRNA expression of steroidogenic enzyme genes in the GCs of hierarchical follicles, but inhibited the effects of FSH in the GCs of prehierarchical follicles. Collectively, these data suggest that Wnt4 plays an important role in chicken follicle selection by stimulating GC proliferation and steroidogenesis. This study provides a theoretical basis for improving the egg-laying performance of chicken and a reference for the elucidation of the molecular mechanism of follicular selection in mammals.

Stem cell pools in the SAM (shoot apical meristem), RAM (root apical meristem) and vascular procambium/cambium are regulated by CLE-receptor kinase-WOX signaling modules. Previous data showed that cyst nematode CLE-like effector proteins delivered into host cells through a stylet, act as ligand mimics of plant A-type CLE peptides and are pivotal for successful parasitism. Here we report the identification of a new class of CLE peptides from cyst nematodes with functional similarity to the B-type CLE peptide TDIF (tracheary element differentiation inhibitory factor) encoded by the CLE41 and CLE44 genes in Arabidopsis. We further demonstrate that the TDIF-TDR (TDIF receptor)-WOX4 pathway, which promotes procambial meristem cell proliferation, is involved in beet cyst nematode Heterodera schachtii parasitism. We observed activation of the TDIF pathway in developing feeding sites, reduced nematode infection in cle41 and tdr-1 wox4-1 mutants, and compromised syncytium size in cle41, tdr-1, wox4-1 and tdr-1 wox4-1 mutants. By qRT-PCR and promoter:GUS analyses, we showed that the expression of WOX4 is decreased in a clv1-101 clv2-101 rpk2-5 mutant, suggesting that WOX4 is a potential downstream target of nematode CLEs. Exogenous treatment with both nematode A-type and B-type CLE peptides induced massive cell proliferation in wild type roots, suggesting that the two types of CLEs may regulate cell proliferation during feeding site formation. These findings highlight an important role of the procambial cell proliferation pathway in cyst nematode feeding site formation.

Spectra derived from ENVISAT Advanced Synthetic Aperture Radar (ASAR) and QuikSCAT near-surface ocean winds are investigated over the North Sea. The two sensors offer a wide range of spatial resolutions, from 600 m to 25 km, with different spatial coverage over the area of interest. This provides a unique opportunity to study the impact of the spatial resolution on the spectral properties of the wind over a wide range of length scales. Initially, a sub-domain in the North Sea is chosen, due to the overlap of 87 wind scenes from both sensors. The impact of the spatial resolution is manifested as an increase in spectral density over similar wavenumber ranges as the spatial resolution increases. The 600-m SAR wind product reveals a range of wavenumbers in which the exchange processes between micro- and meso-scales occur; this range is not captured by the wind products with a resolution of 1.5 km or lower. The lower power levels of coarser resolution wind products, particularly when comparing QuikSCAT to ENVISAT ASAR, strongly suggest that the effective resolution of the wind products should be high enough to resolve the spectral properties. Spectra computed from 87 wind maps are consistent with those obtained from several thousands of samples. Long-term spectra from QuikSCAT show that during the winter, slightly higher energy content is identified compared to the other seasons.