Identification, characterization and hypolipidemic effect of novel peptides in protein hydrolysate from Protaetia brevitarsis larvae.

The proteins were mainly derived from Protaetia brevitarsis larval extracts obtained using two empty intestine methods (traditional static method: TSM or salt immersion stress method: SISM) and extraction solvents (water: W or 50 % water-ethanol: W:E), and the proteins were used as objects to investigate the effect of emptying intestine methods on hypolipidemic peptides. The results revealed that the F-2 fractions of protein hydrolysate had stronger in vitro hypolipidemic activity, with the peptides obtained by SISM possessing a stronger cholesterol micelle solubility inhibition rate, especially in SISM-W:E-P. Moreover, a total of 106 peptides were tentatively identified, among which SISM identified more peptides with an amino acid number < 8. Meanwhile, five novel peptides (YPPFH, YPGFGK, KYPF, SPLPGPR and VPPP) exhibited good hypolipidemic activity in vitro and in vivo, among which YPPFH, VPPP and KYPF had strong inhibitory activities on pancreatic lipase (PL) and cholesteryl esterase (CE), and KYPF, SPLPGPR and VPPP could significantly reduce the TG content in Caenorhabditis elegans. Thus, P. brevitarsis can be developed as a naturally derived hypolipidemic component for the development and application in functional foods.

A Wearable Asynchronous Brain-Computer Interface Based on EEG-EOG Signals With Fewer Channels.

OBJECTIVE: Brain-computer interfaces (BCIs) have tremendous application potential in communication, mechatronic control and rehabilitation. However, existing BCI systems are bulky, expensive and require laborious preparation before use. This study proposes a practical and user-friendly BCI system without compromising performance. METHODS: A hybrid asynchronous BCI system was developed based on an elaborately designed wearable electroencephalography (EEG) amplifier that is compact, easy to use and offers a high signal-to-noise ratio (SNR). The wearable BCI system can detect P300 signals by processing EEG signals from three channels and operates asynchronously by integrating blink detection. RESULT: The wearable EEG amplifier obtains high quality EEG signals and introduces preprocessing capabilities to BCI systems. The wearable BCI system achieves an average accuracy of 94.03±4.65%, an average information transfer rate (ITR) of 31.42±7.39 bits/min and an average false-positive rate (FPR) of 1.78%. CONCLUSION: The experimental results demonstrate the feasibility and practicality of the developed wearable EEG amplifier and BCI system. SIGNIFICANCE: Wearable asynchronous BCI systems with fewer channels are possible, indicating that BCI applications can be transferred from the laboratory to real-world scenarios.

Association of triglyceride-glucose index with the prevalence of cardiovascular disease in malnourished/non-malnourished patients: a large cross-sectional study.

Background: Numerous investigations have demonstrated a strong association between the TyG (triglyceride-glucose) index, which is derived from lipid and glucose levels in the bloodstream, and the onset and progression of cardiovascular diseases (CVD). Blood glucose and blood lipids are affected by nutritional status, and few studies have explored whether the correlation between TyG index and the risk of CVD is affected by nutritional status. Aims: To investigate the connection between TyG index and the risk of CVD among individuals with varying nutritional statuses. Method: A total of 19,847 were included in the analysis, of which 15,955 participants were non-malnourished and 3,892 patients were malnourished. According to the TyG index quartile, the patients were categorized into four groups. Logistic regression analysis and restricted cubic spline was used to study the relationship between TyG index and the risk of CVD in normal and malnourished populations. Results: The results of the restricted cubic spline showed that the TyG index was positively associated with the risk of CVD in the non-malnourished population. The TyG index showed a U-shaped association with the risk of CVD in malnourished people. The result is consistent with that of logistic regression (Malnutrition: Group 2: OR: 1.14; 95% CI: 0.85-1.53; Group 3: OR: 1.36; 95% CI: 1.03-1.79; Group 4: OR: 1.72; 95% CI:1.31-2.25, P for trend <0.001; Non-malnutrition: Group 2: OR: 0.82; 95% CI: 0.46-1.48; Group 3: OR: 0.88; 95% CI: 0.49-1.57; Group 4: OR: 1.45; 95% CI:0.83-2.52, P for trend =0.067). Conclusions: The association between the TyG index and the risk of CVD varied depending on the nutritional states. When using TyG index to assess the risk of CVD, stratification combined with nutritional status helps to more accurately screen patients at high risk of CVD.

Combination therapy with toripalimab and anlotinib in advanced esophageal squamous cell carcinoma: A case report.

BACKGROUND: Toripalimab and anlotinib have shown good response in esophageal cancer, with high objective response rate and progression free survival. Thus, they have been approved as second-line or above-line therapy for advanced or unresectable esophageal carcinoma. Combination of these two drugs may have synergistic effects, but evidence of which is lacking. CASE SUMMARY: Here, we report on a 73-year-old male, newly diagnosed with advanced esophageal squamous cell carcinoma (ESCC), who received a combination of toripalimab and anlotinib. Complete response was achieved after treatment for 3 mo and remission was maintained up to 14 mo. CONCLUSION: The combination therapy of toripalimab and anlotinib is a promising treatment for unresectable ESCC and related clinical trials are warranted.

Lysine 117 Residue Is Essential for the Function of the Hepatocyte Nuclear Factor 1α.

Hepatocyte nuclear factor 1α (HNF1α) plays essential roles in controlling development and metabolism; its mutations are clearly linked to the occurrence of maturity-onset diabetes of the young (MODY3) in humans. Lysine 117 (K117) to glutamic acid (E117) mutation in the HNF1α gene has been clinically associated with MODY3, but no functional data on this variant are available. Here, we addressed the role of lysine 117 in HNF1α function using a knock-in animal model and site-directed mutagenesis. HNF1α K117E homozygous mice exhibited dwarfism, hepatic dysfunction, renal Fanconi syndrome, and progressive wasting syndrome. These phenotypes were very similar to those of mice with complete HNF1α deficiency, suggesting that K117 is critical to HNF1α functions. K117E homozygotes developed diabetes in the early postnatal period. The relative deficiency of serum insulin levels and the normal response to insulin treatment in homozygous mice were markedly similar to those in the MODY3 disorder in humans. Moreover, K117E heterozygous mutant causes age-dependent glucose intolerance, which is similar to the pathogenesis of MODY3 as well. K117 mutants significantly reduced the overall transactivation and DNA binding capacity of HNF1α by disrupting dimerization. Collectively, our findings reveal a previously unappreciated role of POU domain of HNF1α in homodimerization and provide important clues for identifying the molecular basis of HNF1α-related diseases such as MODY3. ARTICLE HIGHLIGHTS: HNF1α K117E homozygous mice exhibited dwarfism, hepatic dysfunction, renal Fanconi syndrome, and progressive wasting syndrome. K117E homozygotes developed diabetes in the early postnatal period. K117E heterozygous mutant causes age-dependent glucose intolerance, which is similar to the pathogenesis of maturity-onset diabetes of the young. K117 mutants significantly reduced the overall transactivation and DNA binding capacity of HNF1α by disrupting dimerization.

Enantioselective Total Syntheses of Grayanane Diterpenoids and (+)-Kalmanol: Evolution of the Bridgehead Carbocation-Based Cyclization and Late-Stage Functional Group Manipulation Strategies.

Grayanane diterpenoids contain over 300 highly oxidized and structurally complex members, many of which possess important biological activities. Full details are provided for the development of the concise, enantioselective and divergent total syntheses of grayanane diterpenoids and (+)-kalmanol. The unique 7-endo-trig cyclization based on a bridgehead carbocation was designed and implemented to construct the 5/7/6/5 tetracyclic skeleton, demonstrating the practical value of the bridgehead carbocation-based cyclization strategy. Extensive studies of late-stage functional group manipulation were performed to forge the C1 stereogenic center, during which a photoexcited intramolecular hydrogen atom transfer reaction was discovered and the mechanism was further studied through density functional theory (DFT) calculations. The biomimetic 1,2-rearrangement from the grayanoid skeleton provided a 5/8/5/5 tetracyclic framework and resulted in the first total synthesis of (+)-kalmanol.

Pro-aromaticity Enabled Dealkenylative Functionalizations via Photo-Excitation and Oxidation.

A general and practical approach for diverse dealkenylative functionalization of olefin-containing substrates has been developed through the one-pot formation and utilization of pro-aromatic 1,4-dihydropyridazines using tetrazine as the key cycloaddition reagent. Triggered by either excitation or oxidation, the targeted C-C bonds in the 1,4-dihydropyridazine intermediates could be readily cleaved to generate alkyl radicals for subsequent transformations. Diverse carbon-carbon and carbon-hetero bond forming protocols, including Giese-type addition, hydrazination, borylation, Minisci-type alkylation, copper-catalyzed NH alkylation, acylation, alkynylation, cyanation, and azidation, are achieved in a highly modular fashion.

Ray-Stretching Statistics and Hot-Spot Formation in Weak Random Disorder.

Weak scattering in a random disordered medium and the associated extreme-event statistics are of great interest in various physical contexts. Here, in the context of non-relativistic particle motion through a weakly correlated random potential, we show how extreme events in particle densities are strongly related to the stretching exponents, where the 'hot spots' in the intensity profile correspond to minima in the stretching exponents. This strong connection is expected to be valid for different random potential distributions, as long as the disorder is correlated and weak, and is also expected to apply to other physical contexts, such as deep ocean waves.

Characterization of bioactives and in vitro biological activity from Protaetia brevitarsis larval extracts obtained by different pretreatment extractions.

Intestinal contents affect the characterization of edible insect bioactive compounds. Two empty intestine methods, namely, traditional static method (TSM) or salt immersion stress method (SISM), associated with extraction solvents water (W), 50 % water-ethanol (W:E) or 100 % ethanol (E), were used to obtain six Protaetia brevitarsis larval extracts. The total flavonoid content (TFC) in the W:E extracts was significantly higher than that in the W and E extracts, with TSM-W:E the highest (p < 0.05). The relative contents of 132 bioactive compounds, especially p-hydroxyphenylacetic acid, citric acid, and dehydroepiandrosterone, were different between TSM-W and SISM-W. TSM-W:E had significantly higher 2,2-diphenyl-1-picrylhydroxy· (DPPH) scavenging and pancreatic lipase (PL) inhibitory activity than SISM-W:E (p < 0.05). DPPH scavenging and PL inhibitory activities were highly correlated with TFC and carbohydrates, respectively. Thus, bioactive compounds in P. brevitarsis extracts can be obtained selectively using pretreatment methods, which might be beneficial for high-value utilization of P. brevitarsis.

The potential of degrading natural chitinous wastes to oligosaccharides by chitinolytic enzymes from two Talaromyces sp. isolated from rotten insects (Hermetia illucens) under solid state fermentation.

It is difficult to produce chitin oligosaccharides by hydrolyzing untreated natural chitinous waste directly. In this study, two fungi Talaromyces allahabadensis Hi-4 and Talaromyces funiculosus Hi-5 from rotten black soldier fly were isolated and identified through multigene phylogenetic and morphological analyses. The chitinolytic enzymes were produced by solid state fermentation, and the growth conditions were optimized by combining single-factor and central composite design. The best carbon sources were powder of molting of mealworms (MMP) and there was no need for additional nitrogen sources in two fungi, then the maximum chitinolytic enzyme production of 46.80 ± 3.30 (Hi-4) and 55.07 ± 2.48 (Hi-5) U/gds were achieved after analyzing the 3D response surface plots. Pure chitin (colloidal chitin) and natural chitinous substrates (represented by MMP) were used to optimize degradation abilities by crude enzymes obtained from the two fungi. The optimum temperature for hydrolyzing MMP (40 °C both in two fungi) were lower and closer to room temperature than colloidal chitin (55 °C for Hi-4 and 45 °C for Hi-5). Then colloidal chitin, MMP and the powder of shrimp shells (SSP) were used for analyzing the products after 5-day degradation. The amounts of chitin oligosaccharides from SSP and MMP were about 1/6 (Hi-4), 1/17 (Hi-5) and 1/8 (Hi-4), 1/10 (Hi-5), respectively, in comparison to colloidal chitin. The main components of the products were GlcNAc for colloidal chitin, (GlcNAc)2 for MMP, and oligosaccharides with higher degree of polymerization (4-6) were obtained when hydrolyzing SSP, which is significant for applications in medicine and health products.

Total Synthesis of (+)-Mutilin: A Transannular [2+2] Cycloaddition/Fragmentation Approach.

A new and enantioselective total synthesis of the diterpenoid (+)-mutilin is described. Following a Claisen rearrangement approach to construct the 6,9-bicycle, a transannular [2+2] photocycloaddition and the ensuing ring-opening reaction were implemented to forge the characteristic 5-6-8 propellane-like skeleton. Subsequent late-stage alkylations and reduction completed the synthesis.

16S rRNA Gene Sequencing Reveals Specific Gut Microbes Common to Medicinal Insects.

Insects have a long history of being used in medicine, with clear primary and secondary functions and less side effects, and the study and exploitation of medicinal insects have received increasing attention. Insects gut microbiota and their metabolites play an important role in protecting the hosts from other potentially harmful microbes, providing nutrients, promoting digestion and degradation, and regulating growth and metabolism of the hosts. However, there are still few studies linking the medicinal values of insects with their gut microbes. In this study, we focused on the specific gut microbiota common to medicinal insects, hoping to trace the potential connection between medicinal values and gut microbes of medicinal insects. Based on 16S rRNA gene sequencing data, we compared the gut microbiota of medicinal insects [Periplaneta americana, Protaetia (Liocola) brevitarsis (Lewis) and Musca domestica], in their medicinal stages, and non-medicinal insects (Hermetia illucens L., Tenebrio molitor, and Drosophila melanogaster), and found that the intestinal microbial richness of medicinal insects was higher, and there were significant differences in the microbial community structure between the two groups. We established a model using a random-forest method to preliminarily screen out several types of gut microbiota common to medicinal insects that may play medicinal values: Parabacteroides goldsteinii, Lactobacillus dextrinicus, Bifidobacterium longum subsp. infantis (B. infantis), and Vagococcus carniphilus. In particular, P. goldsteinii and B. infantis were most probably involved in the anti-inflammatory effects of medicinal insects. Our results revealed an association between medicinal insects and their gut microbes, providing new development directions and possibly potential tools for utilizing microbes to enhance the medicinal efficacy of medicinal insects.

Enhancing nonspecific enzymatic hydrolysis of chitin to oligosaccharides pretreated by acid and green solvents under simultaneous microwave-radiation.

It is hard to degrade untreated highly crystalline chitin. In this study, two solvents pretreatment chitin (acid swollen chitin (AC), super fine chitin (FC)) and microwave-heating method were used to enhance nonspecific enzymatic hydrolysis (lysozyme and pepsin), which obviously improved the enzymolysis rates by at least 1.31 times. Characterizations of chitin substrates (Mv, SEM, XRD) showed that calcium solvent pretreatment (obtained FC) was milder but effective than phosphoric acid pretreatment (obtained AC). The highest yield of chitin oligosaccharides (37.58 mg/g) were obtained after hydrolyzing AC under five-hour simultaneous microwave radiation by pepsin, among them, the content of N-acetylglucosamine was 13.76 mg/g. While, more chitin oligosaccharides with DP (degree of polymerization) 3-4 and lower DA (degree of acetylation) were obtained when using lysozyme than pepsin. Significantly, the conversion rate of chitin to oligosaccharides went best only when microwave and enzymes acting together (simultaneous strategy), which were at least 35.59% higher than separately pretreatment enzymes and substrates by microwave. The damages of microwave radiation on lysozyme and chitin substrates were revealed, and the operating principle of the whole enzyme reaction system heated by microwave was preliminatively explored.

Characterization, Antioxidant Activities, and Pancreatic Lipase Inhibitory Effect of Extract From the Edible Insect Polyrhachis vicina.

Oxidative stress and obesity are critical risk factors for metabolic syndrome. The consumption of functional food ingredients can a viable strategy to alleviate oxidative stress and obesity. In this study, the hydro-ethanolic extract of the edible insect Polyrhachis vicina was prepared and its bioactive components were characterized. The total polyphenol contents, total flavonoid contents, antioxidant and pancreatic lipase (PL) inhibitory activities of the extract were determined in vitro. In total, 60 bioactive components were tentatively identified in the P. vicina extract. Polyphenols and fatty acids were further quantified using LC-MS and GC-MS, respectively. P. vicina extract possessed excellent antioxidant and PL inhibition activities. Salicylic acid, gallic acid, liquiritigenin, and naringenin, which were the major polyphenols in the P. vicina extract, interacted with PL through hydrogen bonding, hydrophilic or hydrophobic and pi-cation interactions. Thus, P. vicina extract can be used as a nutraceutical to alleviate oxidative stress-induced disease and manage obesity.

Scalable Total Synthesis of (-)-Triptonide: Serendipitous Discovery of a Visible-Light-Promoted Olefin Coupling Initiated by Metal-Catalyzed Hydrogen Atom Transfer (MHAT).

An efficient and scalable total synthesis of (-)-triptonide is accomplished based on a metal-catalyzed hydrogen atom transfer (MHAT)-initiated radical cyclization. During the optimization of the key step, we discovered that blue LEDs significantly promoted the efficiency of reaction initiated by Co(TPP)-catalyzed MHAT. Further exploration and optimization of this catalytic system led to development of a dehydrogenative MHAT-initiated Giese reaction.

Dealkenylative Ni-Catalyzed Cross-Coupling Enabled by Tetrazine and Photoexcitation.

A new and general method to functionalize the C(sp3)-C(sp2) bond of alkyl and alkene linkages has been developed, leading to the dealkenylative generation of carbon-centered radicals that can be intercepted to undergo Ni-catalyzed C(sp3)-C(sp2) cross-coupling. This one-pot protocol leverages the easily procured alkene feedstocks for organic synthesis with excellent functional group compatibility without the need for a photoredox catalyst.

Loss of Hepatic Angiotensinogen Attenuates Sepsis-Induced Myocardial Dysfunction.

[Figure: see text].

Disrupted Strength and Stability of Regional Brain Activity in Disorder of Consciousness Patients: A Resting-State Functional Magnetic Resonance Imaging Study.

Although the resting-state networks of patients with disorders of consciousness (DOC) have been widely investigated, the underlying neural mechanisms remain unclear. Here we aimed to explore the static and dynamic alterations in the regional brain activity in patients with DOC and detect the diagnostic ability of each index. Nineteen patients in the vegetative state, 19 in the minimally conscious state (MCS), and 41 healthy controls were recruited for this study. The fractional amplitudes of the low-frequency fluctuation (fALFF) and dynamic fALFF (dfALFF) values were computed, and intergroup differences were detected by analysis of variance. Sub-frequency analysis (slow-4 band and slow-5 band) of fALFF was also performed. Machine learning classifiers were established based on these measures to explore the classification accuracies between patients and controls. The fALFF and dfALFF analyses showed significant intergroup differences in the medial prefrontal gyrus, precuneus, left angular gyrus, and right middle cingulate cortex (MCC), whereas only the dfALFF analysis revealed aberrations in the right inferior frontal gyrus (IFG), right angular gyrus, left supramarginal gyrus (SMG), and left middle occipital gyrus (MOG). Sub-frequency analysis suggested a potential frequency dependent alteration in the default mode network (DMN). The fALFF model exhibited a higher classification accuracy (ACC) (90.50%) than the dfALFF model (86.29%). The combination of fALFF and dfALFF did not improve classification performance. The strength and stability of regional brain activities were disrupted in patients with DOC. Our findings demonstrate that dynamic analysis may reveal more pathological regions and provide a better understanding of the pathophysiologic mechanisms of DOC.

The noncanonical role of the protease cathepsin D as a cofilin phosphatase.

Cathepsin D (cathD) is traditionally regarded as a lysosomal protease that degrades substrates in acidic compartments. Here we report cathD plays an unconventional role as a cofilin phosphatase orchestrating actin remodeling. In neutral pH environments, the cathD precursor directly dephosphorylates and activates the actin-severing protein cofilin independent of its proteolytic activity, whereas mature cathD degrades cofilin in acidic pH conditions. During development, cathD complements the canonical cofilin phosphatase slingshot and regulates the morphogenesis of actin-based structures. Moreover, suppression of cathD phosphatase activity leads to defective actin organization and cytokinesis failure. Our findings identify cathD as a dual-function molecule, whose functional switch is regulated by environmental pH and its maturation state, and reveal a novel regulatory role of cathD in actin-based cellular processes.

Preparation of Antioxidant and Antibacterial Chitosan Film from Periplaneta americana.

Among different insects, the American cockroach (Periplaneta americana) has been bred in industrial scale successfully as a potential resource of protein, lipid, and antibacterial peptide. However, the application of its chitosan has not been studied widely, which has hindered the sufficient utilization of P. americana. In this paper, the chitosan from P. americana was separated, characterized, and processed into film (PaCSF) to examine its potential of being applied in food packaging. As the results of different characterizations showed, PaCSF was similar to shrimp chitosan film (SCSF). However, concerning the performances relating to food packaging, the two chitosan films were different. PaCSF contained more water (42.82%) than SCSF did, resulting in its larger thickness (0.08 mm). PaCSF could resist UV light more effectively than SCSF did. Concerning antioxidant activity, the DPPH radical scavenging ability of PaCSF increased linearly with time passing, reaching 72.46% after 8 h, which was better than that of SCSF. The antibacterial activity assay exhibited that PaCSF resisted the growth of Serratia marcescens and Escherichia coli more effectively than SCSF did. The results implied that P. americana chitosan could be a potential raw material for food packaging, providing a new way to develop P. americana.