Comparison of inducible versus constitutive expression of plectasin on yields and antimicrobial activities in Pichia pastoris.

BACKGROUND: Plectasin might serve as a substitute for traditional antibiotics, but its yields and antimicrobial activities warrant further investigation. OBJECTIVE: To identify the influence of inducible versus constitutive expression of plectasin on yields and antimicrobial activities. METHODS: Through SOE-PCR, a recombinant plectasin gene was generated and inserted into inducible (pPICZαA) and constitutive (pGAPZαA) vectors in order to create Pichia pastoris GS115 strains. After 120 h of fermentation, supernatants were purified by an AKTA purifier using nickel columns. Minimal inhibitory concentration (MIC) and inhibition zone assays were performed after Tricine-SDS-PAGE. RESULTS: After 120 h of fermentation, the yield of constitutive plectasin (370 µg/ml) was much lower than that from inducible vector (880 µg/ml) (P < 0.05). However, constitutive strain reached its plateau phase faster and keep more consistent yield (P < 0.05). The MICs of inducible plectasin against Methicillin-resistant Staphylococcus aureus (MRSA) 15471118, vancomycin-resistant Enterococcus feces (VREF), and penicillin-resistant Streptococcus pneumonia (PRSP) 31355 were 64, 32, and 64 µg/ml, respectively, while those of constitutive plectasin were 4, 4, and 16 µg/ml. No significant differences were observed in antimicrobial activities between inducible and constitutive plectasin for MRSA 15471118, VREF and PRSP 31355 (all P ï¼ž 0.05). However, constitutive plectasin had a larger inhibition zone than inducible plectasin with the same mass. CONCLUSIONS: Although P. pastoris GS115 (pGAPZαA-Plectasin-GS115) had lower expression than P. pastoris GS115 (pPICZαA-plectasin-GS115), it reached the plateau phase faster, had steadier yields and showed superiority in antimicrobial activities. Therefore, pGAPZαA might be more suitable for expression of plectasin in GS115 compared with pPICZαA.

Prognostic Value of Follicle-Stimulating Hormone Levels in Predicting Survival in Men With Idiopathic Pulmonary Arterial Hypertension.

The objective of the study was to assess the association between changes in plasma follicle-stimulating hormone (FSH) and the potential effect on idiopathic pulmonary arterial hypertension (IPAH) in male patients. A total of 116 male patients with IPAH and 53 healthy controls were included from XX Hospital. Plasma FSH concentration was assessed in all participants. Receiver operating characteristic curves were used to assess the mortality risk. Kaplan-Meier curve and Cox regression analyses were used to predict the value of FSH on the survival rate of male IPAH patients. The plasma FSH concentration in the IPAH group was significantly higher than that in the control group (p = .017). Nonsurvivors had significantly higher levels of FSH than survivors (p < .0001). FSH levels were positively correlated with World Health Organization Functional Class, mean pulmonary artery pressure, and pulmonary vascular resistance (PVR; p = .023, p < .0001, and p < .0001, respectively) and negatively correlated with 6-min walk distance (6MWD) and cardiac output (CO; p = .004 and p = .010). Cox regression model analysis showed that the levels of FSH were also the independent factors of mortality in male IPAH patients (p < .0001). The IPAH patients with higher FSH levels had higher PVR, lower 6MWD, CO, and a lower survival rate (p = .042, p = .003, p = .029, and p < .0001, respectively). Therefore, we identified that increased FSH levels were associated with disease severity in male patients with IPAH and independently predicted risk of disease and poor survival rate.

The expression profiling of microRNA in systemic sclerosis-associated pulmonary arterial hypertension.

BACKGROUND: The role of microRNAs (miRNAs) in the pathogenesis of systemic sclerosis-associated pulmonary arterial hypertension (SSc-PAH) remains to be fully elucidated. This study evaluated the expression profile of miRNAs in the lung tissue of patients with SSc-PAH. METHODS: Lung tissue samples were collected from 3 SSc-PAH patients and 4 healthy controls. A small RNA high throughput sequence approach was used for screening the differentially expressed miRNAs in the lung tissue samples. Real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR) was used to validate 4 highly significant differentially expressed miRNAs. Gene Ontology and KEGG (Kyoto Encyclopedia of Genes and Genomes) enrichment analysis for mRNAs were performed using the R package clusterProfiler software. RESULTS: A total of 82 upregulated miRNAs and 35 downregulated miRNAs were detected in the lung tissues of patients with SSc-PAH compared with healthy controls. GO enrichment analysis demonstrated that the upregulated target genes were closely involved in biological processes such as nervous system development, anatomical structure morphogenesis, system development, cellular macromolecule metabolic processes, and cellular processes. The downregulated target genes were involved in the plasma membrane bound cell projection morphogenesis and the regulation of macromolecule metabolic processes. The KEGG enrichment analysis showed that the upregulated genes were associated with important pathways involved in cancer biology, and the target genes of the downregulated miRNAs were involved in axon guidance. High throughput sequencing and qRT-PCR revealed that hsa-miR-205-5p and hsa-miR-539-3p were differentially expressed in SSc-PAH tissue. The target genes of hsa-miR-205-5p and hsa-miR-539-3p, IRF1and ADCYAP1, respectively, were verified using the high throughput dataset GSE48149. CONCLUSIONS: miRNAs may play an important role in the pathogenesis of SSc-PAH, and hsa-miR-205-5p and hsa-miR-539-3p may be potential therapeutic targets in patients with SSc-PAH.

Temporal course of vibrotactile imagery.

This study examined the temporal courses of imagery processes by having participants imagining vibrotactile sensation as if perceived by the right-hand second finger. The results indicate that the imagery condition elicited a less negative going N400 and then a less positive going P600 than the control condition (perceived without imagining). This is suggestive of a sequential process of retrieving experiences from memory, generating images, and then maintaining the images. Dipole analysis further supports that N400 and P600 (or long latency P300) can be respectively regarded as part of the generic 'imagery network' and as modality-specific processes of imagery.

Attention shortage resistance of negative stimuli in an implicit emotional task.

Considerable evidence from behavioral studies has indicated that people tend to pay attention to negative stimuli preferentially. The attentional bias can occur rapidly and automatically. In the current study, a 'cue-target' paradigm was utilized to manipulate the attention allocation. Seventeen healthy undergraduates participated in the experiment. The stimuli were emotional pictures (positive, neutral and negative), which were upper and lower adjacent patchworks of a normal scene and its inverted copy. The subjects should judge whether the normal scene (compared with the inverted scene) was located in the upper or lower part of the whole patchwork. We used this implicit emotional task to avoid the task relevance effect. It was found that the amplitude of P2 waves was enlarged by the negative pictures and there was a significant interaction between the cue effect and the emotional valence. We can conclude that the negative information exerts an attentional bias effect in the emotional perception, and that the negative contents suffer less in the insufficient attention condition compared with the positive and the neutral conditions.

Temporal course of emotional negativity bias: an ERP study.

There is considerable evidence that people are especially sensitive to emotionally negative materials. However, the temporal course of the negativity bias is still unclear. To address this issue, we observed the changes of P2, late positive components (LPC) and lateralized readiness potential (LRP) under positive, negative and neutral conditions, with International Affective Picture System (IAPS) pictures as emotional stimuli. We found that the amplitude of P2 in the negative block was significantly larger than that in the positive block, indicating that the attentional negativity bias occured very early in emotion perception. The LPC amplitude evoked by negative pictures was larger than that by positive and neutral pictures, suggesting that the negativity bias also occurred in a later evaluation stage of emotion processing. The response-locked LRP interval was shortest in the block of negative pictures, indicating that the negative contents elicited a reaction priming effect. Above all, this research showed that emotional negativity bias could occur in several temporal stages distinguished by attention, evaluation and reaction readiness periods.

JAK/STAT signaling pathway-mediated immune response in silkworm (Bombyx mori) challenged by Beauveria bassiana.

Innate immunity was critical in insects defensive system and able to be induced by Janus kinase/signal transducer and activator of transcription cascade transduction (JAK/STAT) signaling pathway. Currently, it had been identified many JAK/STAT signaling pathway-related genes in silkworm, but little function was known on insect innate immunity. To explore the roles of JAK/STAT pathway in antifungal immune response in silkworm (Bombyx mori) against Beauveria bassiana infection, the expression patterns of B. mori C-type lectin 5 (BmCTL5) and genes encoding 6 components of JAK/STAT signaling pathway in silkworm challenged by B. bassiana were analyzed using quantitative real time PCR. Meanwhile the activation of JAK/STAT signaling pathway by various pathogenic micro-organisms and the affect of JAK/STAT signaling pathway inhibitors on antifungal activity in silkworm hemolymph was also detected. Moreover, RNAi assay of BmCTL5 and the affect on expression levels of signaling factors were also analyzed. We found that JAK/STAT pathway could be obviously activated in silkworm challenged with B. bassiana and had no response to bacteria and B. mori cytoplasmic polyhedrosis virus (BmCPV). However, the temporal expression patterns of JAK/STAT signaling pathway related genes were significantly different. B. mori downstream receptor kinase (BmDRK) might be a positive regulator of JAK/STAT signaling pathway in silkworm against B. bassiana infection. Moreover, antifungal activity assay showed that the suppression of JAK/STAT signaling pathway by inhibitors could significantly inhibit the antifungal activity in hemolymph and resulted in increased sensitivity of silkworm to B. bassiana infection, indicating that JAK/STAT signaling pathway might be involved in the synthesis and secretion of antifungal substances. The results of RNAi assays suggested that BmCTL5 might be one pattern recognition receptors for JAK/STAT signaling pathway in silkworm. These findings yield insights for better understand the molecular mechanisms of JAK/STAT signaling pathway in antifungal immune response in silkworm.

Efficiency of neural respiratory drive for the assessment of bronchodilator responsiveness in patients with chronic obstructive pulmonary disease: an exploratory study.

BACKGROUND: Conventional lung function parameters, such as forced vital capacity (FVC), forced expiratory volume in 1 second (FEV1) and inspiratory capacity (IC) are often used to assess the therapeutic outcomes of bronchodilators, but they lack sensitivity. A novel indicator, namely efficiency of neural respiratory drive (NRD), may objectively evaluate the physiological changes in patients with chronic obstructive pulmonary disease (COPD). We investigated whether this indicator could be used to more accurately assess the responsiveness to inhaled bronchodilators. METHODS: Thirty-six subjects with moderate-to-severe COPD were randomized into group A (n=18) and group B (n=18). Participants in group A inhaled 400 µg placebo, 400 µg salbutamol and 80 µg ipratropium in sequence whereas those in group B had the salbutamol and ipratropium reversed. At different time points after administration of placebo or bronchodilators, evaluated indices included FEV1, FVC, IC, root mean square (RMS) of diaphragm electromyogram (EMGdi), and efficiency of NRD [herein defined as the ratio of minute ventilation (VE) to RMS, or VE/RMS]. RESULTS: FEV1, FVC, IC, RMS, and VE/RMS significantly improved after inhaled bronchodilators and VE/RMS had the largest improvement among five indices. The detection efficiency of VE/RMS was greater than FEV1, FVC, IC (all P<0.05), but not different from RMS. The accuracy and sensitivity of VE/RMS were significantly higher than FEV1, FVC, IC, and RMS (all P<0.05). CONCLUSIONS: Efficiency of NRD may be a sensitive tool to evaluate the efficacy of inhaled bronchodilators in COPD.

The delay effect on outcome evaluation: results from an event-related potential study.

Behavioral studies demonstrate that the timing of receiving gains or losses affects decision-making, a phenomenon known as temporal discounting, as participants are inclined to prefer immediate rewards over delayed ones and vice versa for losses. The present study used the event-related potential technique with a simple gambling task to investigate how delayed rewards and losses affected the brain activity in outcome evaluations made by 20 young adults. Statistical analysis revealed a larger feedback-related negativity (FRN) effect between loss and gain following immediate outcomes than following future outcomes. In addition, delay impacted FRN only in gain conditions, with delayed winning eliciting a more negative FRN than immediate winning. These results suggest that temporal discounting and sign effect could be encoded in the FRN in the early stage of outcome evaluation.

Cross-brain neurofeedback: scientific concept and experimental platform.

The present study described a new type of multi-person neurofeedback with the neural synchronization between two participants as the direct regulating target, termed as "cross-brain neurofeedback." As a first step to implement this concept, an experimental platform was built on the basis of functional near-infrared spectroscopy, and was validated with a two-person neurofeedback experiment. This novel concept as well as the experimental platform established a framework for investigation of the relationship between multiple participants' cross-brain neural synchronization and their social behaviors, which could provide new insight into the neural substrate of human social interactions.

[Study on the bioleaching mechanism of manganse (II) from manganese-electrolytic residue by manganese-resistant strain Fusarium sp].

The manganse bioleaching mechanism by a manganese-resistant strain Fusarium sp. was investigated, through analyzing the bioleaching rate and manganese-electrolytic residue characterizations with the presence of Fusarium sp. and with the addition of organic acids. Special attention was paid to explore the relationship among the manganese's leaching rate, pH, and organic acid concentration during Fusarium sp. bioleaching process. The research results showed that, with the addition of Fusarium sp., some looser and more porous manganese-electrolytic residues could be obtained. And after 47 hours, the leaching rate reached to 84% which was 2.30 times higher than that leached by individual organic acid even after 130 hours; the leaching rate of manganese and the concentrations of organic acids increased at the initial stage and then decreased, while pH was the reversed. Additionally, the concentration of Succinic acid and L-Malic acid reached their crest value (11.12 g/L and 10.23 g/L) at 57 and 62 hours respectively. Yet the pH reached the lowest (4.09) at 29 h, which implied that, Fusarium sp. and organic acid produced played an important role in the leaching of manganese, leading to a high-efficiency and time-saving process. However, due to the high density of manganese-electrolytic residue and the concurrence of the produce and consumption of organic acid together with the adsorption and complexation, the relationship among the extraction rate for manganese ion, pH, and the concentration of organic acid produced could not be described by simple linear correlation and the leaching rate decreased significantly in the later stage.

Anxiety and feedback negativity.

It has been suggested that anxious individuals are more prone to feel that negative outcomes are particularly extreme and to interpret ambiguous outcomes as negative compared to nonanxious individuals. Previous studies have demonstrated that the feedback negativity (FN) component of event-related brain potential (ERP) is sensitive to outcome evaluation and outcome expectancy. Hence, we predicted that the FN should be different between high trait-anxiety (HTA) and low trait-anxiety (LTA) individuals. To test our hypothesis, the ERPs were recorded during a simple monetary gambling task. The FN was measured as a difference wave created across conditions. We found that the amplitude of the FN indicating negative versus positive outcomes was significantly larger for LTA individuals compared to HTA individuals. However, there was no significant difference in the FN between groups in response to ambiguous versus positive outcomes. The results indicate that there is a relationship between the FN and individual differences in anxiety. We suggest that these results reflect the impact of anxiety on outcome expectation. Our results challenge the reinforcement learning theory of error-related negativity, which proposes that ERN and FN reflect the same cognitive process.

Influence of trait-anxiety on inhibition function: evidence from ERPs study.

Patients suffering from anxiety disorder may experience a few problems in the inhibition function. Using event-related potentials, the current study investigated the differences between subjects with high versus low trait-anxiety when they tried to inhibit disturbances in novel emotional pictures in an oddball task. The results showed that P3 amplitudes evoked by negative pictures relative to neutral pictures were decreased in subjects with high as well as low anxiety. In the high-anxious group, P3 amplitudes were also decreased in the positive condition relative to the neutral condition, whereas in the low anxious group, P3 amplitudes showed no significant differences between the positive and neutral stimuli. This implies that people with high anxiety may exhibit some degree of over-inhibition in emotional processing as compared to people with low anxiety. These people tend to indiscriminately inhibit all types of disturbing emotional information.