Uncertainty quantification of microcirculatory characteristic parameters for recognition of cardiovascular diseases.

BACKGROUND: Cardiovascular disease is one of the leading causes of death worldwide. However, according to studies, 90% of cardiovascular diseases can be prevented. Cardiovascular function parameters are an important basis for the diagnosis of cardiovascular diseases. The pulse wave also contains a wealth of physiological and pathological information, which can reflect the trend of cardiac function parameters at an early stage, so the measurement and analysis of the pulse wave signal becomes more and more important. The wearable pulse signal acquisition device has gradually become a new trend. In the mobile health scenario, convenient use is the prerequisite for long-term and rapid health monitoring. The data containing diverse pulse wave signals is the basis for obtaining more comprehensive and accurate human physiopathological information. Accurate data analysis and processing is the key to realizing the important goal of cardiovascular health monitoring. OBJECTIVE: Based on the concept of mobile health care, wearable devices are developed to obtain physiological signals. The zero-dimensional model and the optimization algorithm are combined to complete the uncertainty quantification of the microcirculation parameters. Then, a feature set containing the cardiovasvular parameters can be constructed. The machine learning algorithm can be used to build a model that can accurately realize cardiovascular disease identification. METHODS: This paper adopts laboratory-developed equipment to acquire the wrist pulse wave and fingertip volume pulse wave. A total of 323 samples were collected from healthy populations, hypertensive patients and patients with coronary heart disease (CHD). The pulse blood flow model in fingertip microcirculation is established, and the uncertainty quantification of model parameters is completed based on slime mold algorithm (SMA). After comparing and analyzing the performance of four algorithms on pulse wave classification, the identification model of cardiovascular diseases is established based on the microcirculatory characteristic parameter set and random forest algorithm (RF). RESULTS: RF showed good classification performance among the four classification algorithms. The identification accuracy of the model built on the microcirculatory characteristic parameter set and RF algorithm all reached more than 88%. The highest recognition accuracy was 95.51% for coronary heart disease samples, 92.11% for healthy samples, and 88.55% for hypertensive samples. It can be seen that the model based on RF algorithm has a good ability to distinguish the characteristic parameters in different cardiovascular health states. CONCLUSIONS: The wearable device designed in this paper can facilitate the daily health monitoring of cardiovascular disease. By using the combination of the physical model and machine learning model, the uncertainty quantification of microcirculation parameters and the identification of cardiovascular disease was finally completed. The recognition model based on machine learning provides a new idea and method for the research of cardiovascular health monitoring through pulse waves.

Automated Segmentation of the Systolic and Diastolic Phases in Wrist Pulse Signal Using Long Short-Term Memory Network.

Purpose: Single-period segmentation is one of the important steps in time-domain analysis of pulse signals, which is the basis of time-domain feature extraction. The existing single-period segmentation methods have the disadvantages of generalization, reliability, and robustness. Method: This paper proposed a period segmentation method of pulse signals based on long short-term memory (LSTM) network. The preprocessing was performed to remove noises and baseline drift of pulse signals. Thus, LabelMe was used to label each period of the pulse signals into two parts according to the location of the starting point of main wave and the dicrotic notch, and the dataset of the pulse signal period segmentation was established. Consequently, the labeled dataset was input into the LSTM for training and testing, and the results were compared with sum slope function method. Result: The remarkable result with the whole period segmentation accuracy of 92.8% was achieved for the segmentation of seven types of pulse signals. And the segmentation accuracies of the systolic phase, diastolic phase, and whole period using this method were higher than those of the sum slope function method. Conclusion: LSTM-based pulse signal segmentation method can achieve outstanding, robust, and reliable segmentation effects of the systolic phase, diastolic phase, and whole period of pulse signals. The research provides a new idea and method for the segmentation of pulse signals.

Knockdown of hexokinase in Diaphorina citri Kuwayama (Hemiptera: Liviidae) by RNAi inhibits chitin synthesis and leads to abnormal phenotypes.

BACKGROUND: Silencing specific genes in pests using RNA interference (RNAi) technology is a promising new pest-control strategy. The Asian citrus psyllid, Diaphorina citri Kuwayama, is the most important citrus pest because it transmits Candidatus Liberibacter asiaticus, which causes huanglongbing. Chitin is essential for insect development, and enzymes in this pathway are attractive targets for pest control. RESULTS: The hexokinase gene DcHK was characterized from D. citri to impair proper growth and chitin synthesis through RNAi. The transcription of DcHK was more highly developed in third-instar nymphs, adults and the Malpighian tube. The RNAi needed for D. citri is dose-dependent, with 600 ng µl-1 dsDcHK sufficient to knockdown endogenous DcHK expression. The messenger RNA (mRNA) level was lowest at 36 h after dosing, and there were significant effects on the relative levels of mRNA in the chitin synthesis pathway (DcTre, DcG6PI, DcGNAT, DcGFAT, DcPGM, DcUAP and DcCHS), leading to mortality, reduced body weight and abnormal or lethal phenotypes. CONCLUSION: RNAi can be triggered by orally delivered double-stranded RNA in D. citri. These results can provide support for HK genes as a new potential target for citrus psyllid control. © 2022 Society of Chemical Industry.

Genomic characterization of polyextremotolerant black yeasts isolated from food and food production environments.

Black yeasts have been isolated from acidic, low water activity, and thermally processed foods as well as from surfaces in food manufacturing plants. The genomic basis for their relative tolerance to food-relevant environmental stresses has not been well defined. In this study, we performed whole genome sequencing (WGS) on seven black yeast strains including Aureobasidium (n=5) and Exophiala (n=2) which were isolated from food or food production environments. These strains were previously characterized for their tolerance to heat, hyperosmotic pressure, high pressure processing, hypochlorite sanitizers, and ultraviolet light. Based on the WGS data, three of the strains previously identified as A. pullulans were reassigned as A. melanogenum. Both haploid and diploid A. melanogenum strains were identified in this collection. Single-locus phylogenies based on beta tubulin, RNA polymerase II, or translation elongation factor protein sequences were compared to the phylogeny produced through SNP analysis, revealing that duplication of the fungal genome in diploid strains complicates the use of single-locus phylogenetics. There was not a strong association between phylogeny and either environmental source or stress tolerance phenotype, nor were trends in the copy numbers of stress-related genes associated with extremotolerance within this collection. While there were obvious differences between the genera, the heterogenous distribution of stress tolerance phenotypes and genotypes suggests that food-relevant black yeasts may be ubiquitous rather than specialists associated with particular ecological niches. However, further evaluation of additional strains and the potential impact of gene sequence modification is necessary to confirm these findings.

Thermoresistance in Black Yeasts Is Associated with Halosensitivity and High Pressure Processing Tolerance but Not with UV Tolerance or Sanitizer Tolerance.

ABSTRACT: Black yeasts can survive extreme conditions in food production because of their polyextremotolerant character. However, significant strain-to-strain variation in black yeast thermoresistance has been observed. In this study, we assessed the variability in tolerance to nonthermal interventions among a collection of food-related black yeast strains. Variation in tolerance to UV light treatment, high pressure processing (HPP), sanitizers, and osmotic pressure was observed within each species. The two strains previously shown to possess high thermotolerance, Exophiala phaeomuriformis FSL-E2-0572 and Exophiala dermatitidis YB-734, were also the most HPP tolerant but were the least halotolerant. Meanwhile, Aureobasidium pullulans FSL-E2-0290 was the most UV and sanitizer tolerant but had been shown to have relatively low thermoresistance. Fisher's exact tests showed that thermoresistance in black yeasts was associated with HPP tolerance and inversely with halotolerance, but no association was found with UV tolerance or sanitizer tolerance. Collectively, the relative stress tolerance among strains varied across interventions. Given this variation, different food products are susceptible to black yeast spoilage. In addition, different strains should be selected in challenge studies specific to the intervention.

The Primary Complete Mitochondrial Genome of the Lappet Moth Brahmophthalma hearseyi (Lepidoptera: Brahmaeidae) and Related Phylogenetic Analysis.

Background: Brahmophthalma hearseyi (Lepidoptera: Brahmaeidae) is widely distributed across China. Its larvae damage the leaves of many plants such as those belonging to the Oleaceae family, causing significant economic losses and seriously affecting the survival and reproduction of Cervus nippon; however, genetic data for this species are scarce. Methods: The complete mitochondrial genome (mitogenome) of B. hearseyi was sequenced using long-PCR and primer-walking methods. Phylogenetic analysis that was based on 13 PCGs and two rRNAs was carried out using the neighbor-joining and Bayesian interference methods. Results: The mitogenome is a typical circular molecule that is made up of 15,442 bp, which includes 13 protein-coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, 22 transfer RNA (tRNA) genes, and an A + T-rich region (456 bp). All of the PCGs, except for COX1 and COX2, start with ATN codons. COX2 and ND5 use the incomplete termination codon T, and 11 other PCGs use the typical stop codon TAA. All tRNA genes, except for trnS1 and trnS2, display a typical cloverleaf structure; trnS1 lacks the "DHU" arm, whereas trnS2 exhibits two mismatched base pairs in the anticodon stem. Phylogenetic analysis showed that B. hearseyi is clustered into Brahmaeidae, and the phylogenetic relationships are (Brahmaeidae + Lasiocampidae) + (Bombycidae + (Sphingidae + Saturniidae)). Conclusions: This study provides the first mitogenomic resources for the Brahmaeidae.

BEZ235 Increases the Sensitivity of Hepatocellular Carcinoma to Sorafenib by Inhibiting PI3K/AKT/mTOR and Inducing Autophagy.

Acquired resistance of hepatocellular carcinoma (HCC) to sorafenib (SFB) is the main reason for the failure of SFB treatment of the cancer. Abnormal activation of the PI3K/AKT/mTOR pathway is important in the acquired resistance of SFB. Therefore, we investigated whether BEZ235 (BEZ) could reverse acquired sorafenib resistance by targeting the PI3K/mTOR pathway. A sorafenib-resistant HCC cell line Huh7R was established. MTT assay, clone formation assay, flow cytometry, and immunofluorescence were used to analyze the effects of BEZ235 alone or combined with sorafenib on cell proliferation, cell cycle, apoptosis, and autophagy of Huh7 and Huh7R cells. The antitumor effect was evaluated in animal models of Huh7R xenografts in vivo. Western blot was used to detect protein levels of the PI3K/AKT/mTOR pathway and related effector molecules. In vitro results showed that the Huh7R had a stronger proliferation ability and antiapoptosis effect than did Huh7, and sorafenib had no inhibitory effect on Huh7R. SFB + BEZ inhibited the activation of the PI3K/AKT/mTOR pathway caused by sorafenib. Moreover, SFB + BEZ inhibited the proliferation and cloning ability, blocked the cell cycle in the G0/G1 phase, and promoted apoptosis in the two cell lines. The autophagy level in Huh7R cells was higher than in Huh7 cells, and BEZ or SFB + BEZ further promoted autophagy in the two cell lines. In vivo, SFB + BEZ inhibited tumor growth by inducing apoptosis and autophagy. We concluded that BEZ235 enhanced the sensitivity of sorafenib through suppressing the PI3K/AKT/mTOR pathway and inducing autophagy. These observations may provide the experimental basis for sorafenib combined with BEZ235 in trial treatment of HCC.

Intermediate Thermoresistance in Black Yeast Asexual Cells Variably Increases with Culture Age, Promoting Survival and Spoilage in Thermally Processed Shelf-Stable Foods.

ABSTRACT: Black yeasts are a functional group that has caused spoilage in cold-filled and hot-filled beverages, as well as other water activity-controlled food products. We established quantitative thermoresistance parameters for the inactivation of 12 Aureobasidium and Exophiala isolates through isothermal experiments and a challenge study. Culture age (2 versus 28 days) variably affected the thermoresisitance among the black yeast strains. Variation in thermoresistance exists within each genus, but the two most resistant strains were the Exophiala isolates. The two most heat-resistant isolates were Exophiala phaeomuriformis FSL-E2-0572, with a D60-value of 7.69 ± 0.63 min in 28-day culture and Exophiala dermatitidis YB-734, with a D60-value of 16.32 ± 2.13 min in 28-day culture. Although these thermoresistance levels were, in some cases, greater than those for conidia and vegetative cells from other common food spoilage fungi, they were much more sensitive than the ascospores of heat-resistant molds most associated with spoilage of hot-filled products. However, given that black yeasts have caused spoilage in hot-filled products, we hypothesized that this intermediate degree of thermoresistance may support survival following introduction during active cooling before package seals have formed. A challenge study was performed in an acidic (apple cider) and water activity-controlled (maple syrup) product to evaluate survival. When apple cider was hot filled at 82°C, black yeast counts were reduced by 4.1 log CFU/mL 24 h after the heat treatment, but the survivors increased up to 6.7 log CFU/mL after 2 weeks. In comparison, the counts were below the detection limit after both 24 h and 14 days of shelf life in both products when filled at the boiling points. This suggests that ensuring water microbial quality in cooling tunnels and nozzle sanitation may be essential in mitigating the introduction of these fungi.

All Treatment Parameters Affect Environmental Surface Sanitation Efficacy, but Their Relative Importance Depends on the Microbial Target.

Environmental sanitation in food manufacturing plants promotes food safety and product microbial quality. However, the development of experimental models remains a challenge due to the complex nature of commercial cleaning processes, which include spraying water and sanitizer on equipment and structural surfaces within manufacturing space. Although simple in execution, the physical driving forces are difficult to simulate in a controlled laboratory environment. Here, we present a bench-scale bioreactor system which mimics the flow conditions in environmental sanitation programs. We applied computational fluid dynamic (CFD) simulations to obtain fluid flow parameters that better approximate and predict industrial outcomes. According to the CFD model, the local wall shear stress achieved on the target surface ranged from 0.015 to 5.00 Pa. Sanitation efficacy on six types of environmental surface materials (hydrophobicity, 57.59 to 88.61°; roughness, 2.2 to 11.9 µm) against two different microbial targets, the bacterial pathogen Listeria monocytogenes and Exophiala species spoilage fungi, were evaluated using the bench-scale bioreactor system. The relative reduction ranged from 0.0 to 0.82 for Exophiala spp., which corresponded to a 0.0 to 2.21 log CFU/coupon reduction, and the relative reduction ranged from 0.0 to 0.93 in L. monocytogenes which corresponded to a 0.0 to 6.19 log CFU/coupon reduction. Although most treatment parameters were considered statistically significant against either L. monocytogenes or Exophiala spp., contact time was ranked as the most important predictor for L. monocytogenes reduction. Shear stress contributed the most to Exophiala spp. removal on stainless steel and Buna-N rubber, while contact time was the most important factor on HDPE (high-density polyethylene), cement, and epoxy.IMPORTANCE Commercial food manufacturers commonly employ a single sanitation program that addresses both bacterial pathogen and fungal spoilage microbiota, despite the fact that the two microbial targets respond differently to various environmental sanitation conditions. Comparison of outcome-based clusters of treatment combinations may facilitate the development of compensatory sanitation regimes where longer contact time or greater force are applied so that lower sanitizer concentrations can be used. Determination of microbiological outcomes related to sanitation program efficacy against a panel of treatment conditions allows food processors to balance tradeoffs between quality and safety with cost and waste stream management, as appropriate for their facility.

Anti-GPC3 Antibody-Conjugated BEZ235 Loaded Polymeric Nanoparticles (Ab-BEZ235-NP) Enhances Radiosensitivity in Hepatocellular Carcinoma Cells by Inhibition of DNA Double-Strand Break Repair.

AIM: To assess AB-BEZ235-NP potential as a radio-sensitizer in hepatocellular carcinoma models. METHOD: By comparing hepatocellular carcinoma cell with simple radiation or combined AB-BEZ235-NP therapy, the HCC apoptosis and self-repair level have significant differences in mortality rates and cell migration abilities. RESULTS: Cell proliferation and DNA damage increased by pretreatment with AB-BEZ235-NP after irradiation; further studies on the repair pathway indicated that AB-BEZ235-NP inhibited the important pathway of DSB repair. Our results further show that AB-BEZ235-NP significantly inhibits the phosphorylation of the canonical protein, γ-H2AX, in the NHEJ DSB repair pathway and Serine Protein Kinase (SPK) ATM, and TP53-Binding Protein one. More importantly, AB-BEZ235-NP increased the mount of mean γ-H2AX Foci in irradiated cells, indicating that AB-BEZ235-NP can selectively inhibit DSB repair in HCC cells. Therefore, these results clearly eludicate that treatment with AB-BEZ235-NP is a potential promising therapy which can increase the radiosensitivity to HCC.

Resistance of hepatocellular carcinoma to sorafenib can be overcome with co-delivery of PI3K/mTOR inhibitor BEZ235 and sorafenib in nanoparticles.

Background: The combination of BEZ235 with sorafenib (SFB) enhances anti-hepatocellular carcinoma (HCC) efficacy of the two agents. However, pharmacokinetic profiles in vivo and different endocytosis abilities of these two drugs hinder their therapeutic application.Research design and methods: In this work, we developed d-α-tocopheryl polyethylene glycol 1000 succinate - polycaprolactone polymer nanoparticles (NPs) for co-delivery of SFB and BEZ235 (SFB/BEZ235-NPs). Explored the anti-proliferative and pro-apoptotic effects of SFB/BEZ235-NPs through in vitro and in vivo experiments.Results: Stabilized SFB/BEZ235-NPs were prepared with optimized drug ratio, yielding high encapsulation efficiency, low polydispersity, and enhanced cellular internalization in HepG2 cells. Synergistic cytotoxicity and pro-apoptotic ability were documented. In vivo pharmacokinetic results revealed extended circulation and bioavailability of SFB/BEZ235-NPs compared with those of free drugs. SFB/BEZ235-NPs enhanced antitumor effectiveness in SFB-resistant HCC xenograft mouse models.Conclusion: Taken together, the results of this study describe a promising strategy using SFB and BEZ235 in a nanoparticle formulation for treatment of SFB-resistant HCC.

C-reactive protein/serum amyloid P promotes pro-inflammatory function and induces M1-type polarization of monocytes/macrophages in mudskipper, Boleophthalmus pectinirostris.

C-reactive protein (CRP) and serum amyloid P (SAP) play essential roles in the phagocytic cell-mediated innate immune response of mammals. In-depth studies into CRP and SAP have been completed in mammals; however, such studies, particularly those relating to the functions of CRP and SAP, are rare in fish species. In this study, a homolog of CRP/SAP (BpCRP/SAP) was identified in mudskipper (Boleophthalmus pectinirostris), which had the typical characteristics of a fish short pentraxin protein. Phylogenetic tree analysis revealed that BpCRP/SAP was most closely related to mudskipper CRP/SAP-l3. BpCRP/SAP transcripts were detected in all tested tissues, with the highest level observed in the liver; transcripts in the immune tissues and protein expression in the serum were induced in response to Edwardsiella tarda infection. The active recombinant BpCRP/SAP (rBpCRP/SAP) was able to augment the mRNA expression of pro-inflammatory cytokines and attenuate the mRNA expression of anti-inflammatory cytokines in monocytes/macrophages (MO/MΦ). In addition, phagocytosis and bacterial killing of E. tarda by mudskipper MO/MΦ were boosted by rBpCRP/SAP stimulation. rBpCRP/SAP also promoted M1-type MO/MΦ polarization, but inhibited M2-type polarization. In conclusion, the present research describes the pro-inflammatory function of BpCRP/SAP in mudskipper against E. tarda infection.

PKI-587 enhances chemosensitivity of oxaliplatin in hepatocellular carcinoma through suppressing DNA damage repair pathway (NHEJ and HR) and PI3K/AKT/mTOR pathway.

Oxaliplatin resistance limits its effectiveness in the treatment of hepatocellular carcinoma (HCC). Abnormal activation of the PI3K/AKT/mTOR pathway has been associated with decreased survival of HCC patients, anti-apoptosis after chemotherapeutic drug-induced DNA damage, and chemoresistance. In this research, we evaluated the effect of the dual PI3K/mTOR inhibitor, PKI-587, on the sensitivity of oxaliplatin in HCC. Two HCC cell lines (HepG2 and SK-Hep1) were used to analyze PKI-587 for DNA damage response, cell proliferation, clonogenic survival, cell cycle and apoptosis after oxaliplatin treatment. A HepG2 tumor-bearing model was used to assess the in vivo effects of the combination of the two compounds. In HCC cells, oxaliplatin stably activated the PI3K/AKT/mTOR pathway, including up-regulation of p-Akt (Ser473), p-mTOR (Ser2448), p-mTOR (Ser2481), p-elF4EBP1, and p-S6K1, and activated the DNA damage repair pathways (non-homologous end joining (NHEJ) and homologous recombination (HR)), up-regulation of p-DNAPKcs (Ser2056), p-ATM (Ser1981), and p-ATR (Ser428), which were attenuated by PKI-587. Compared with oxaliplatin alone, the combination of PKI-587 and oxaliplatin increased the number of γ-H2AX/cells, decreased proliferation of cells, and an increased the percentage of G0/G1 phase cells and apoptotic cells. In vivo, the combination of oxaliplatin with PKI-587 inhibited tumor growth. Anti-tumor effects were associated with induction of mitochondrial apoptosis and inhibition of phosphorylation of mTOR, Akt and γ-H2AX. We conclude that PKI-587 enhances chemosensitivity of oxaliplatin in HCC through suppressing the PI3K/AKT/mTOR signalling pathway and inhibiting the DNA damage repair pathway. The combination of PKI-587 and oxaliplatin appears to be a promising regimen for the treatment of HCC.

Combined Effect of Storage Condition, Surface Integrity, and Length of Shelf Life on the Growth of Listeria monocytogenes and Spoilage Microbiota on Refrigerated Ready-to-Eat Products.

Psychrotolerant growth of Listeria monocytogenes in ready-to-eat (RTE) foods increases the risk to food safety, particularly when spoilage does not occur prior to L. monocytogenes growth of >1 log CFU/g. The purpose of this study was to evaluate the relative rates of quality deterioration and L. monocytogenes growth in six product systems (tomatoes, apples, fresh-cut cantaloupe, fresh-cut lettuce, baby spinach, and commercially processed turkey slices) under various conditions of refrigeration temperatures, atmospheres, and quality. Cantaloupe and spinach leaves supported >1 log CFU/g growth of L. monocytogenes before product spoilage at both 4 and 9°C. In some cases, conditions that improved microbial quality by extending shelf life also allowed L. monocytogenes growth of >1 log CFU/g before deterioration due to microbial spoilage. For example, storage with modified atmosphere packaging enhanced L. monocytogenes growth relative to spoilage microbiota in lettuce leaves (1.0-log increase 7 days before spoilage). In contrast, the use of secondary quality produce (i.e., apples, tomatoes, and lettuce with physical damage) reduced shelf life and, consequently, limited the time for L. monocytogenes proliferation. Therefore, spoilage cannot be considered a fail-safe indicator or proxy for limitation of shelf life across refrigerated RTE products.

MicroRNA-155 promotes pro-inflammatory functions and augments apoptosis of monocytes/macrophages during Vibrio anguillarum infection in ayu, Plecoglossus altivelis.

Upon recognition of pathogen-associated molecular patterns by pattern-recognition receptors, immune cells are recruited, and multiple antibacterial/viral signaling pathways are activated, leading to the production of immune-related cytokines, chemokines, and interferons along with further activation of the adaptive immune response. MicroRNAs (miRs) play essential roles in regulating such immune signaling pathways, as well as the biological activities of immune cells; however, knowledge regarding the roles of miRs in the immune-related function of monocytes/macrophages (MO/MΦ) remains limited in teleosts. In the present study, we addressed the effects of miR-155 on Vibrio anguillarum-infected MO/MΦ. Our results showed that miR-155 augmented MO/MΦ expression of proinflammatory cytokines and attenuated the expression of anti-inflammatory cytokines. Additionally, the phagocytosis and bacteria-killing abilities of these cells were boosted by miR-155 administration, which also promoted M1-type polarization but inhibited M2-type polarization. Furthermore, the V. anguillarum-infection-induced apoptosis was also enhanced by miR-155 mimic transfection, which might have been due to excessive inflammation or the accumulation of reactive oxygen species. These results represent the first report providing a detailed account of the regulatory roles of miR-155 on MO/MΦ functions in teleosts and offer insight into the evolutionary history of miR-155-mediated regulation of host immune responses.

Involvement of ayu NOD2 in NF-κB and MAPK signaling pathways: Insights into functional conservation of NOD2 in antibacterial innate immunity.

Nucleotide oligomerization domain 2 (NOD2) is a major cytoplasmic sensor for pathogens and is critical for the clearance of cytosolic bacteria in mammals. However, studies regarding NOD2, especially the initiated signaling pathways, are scarce in teleost species. In this study, we identified a NOD2 molecule (PaNOD2) from ayu (Plecoglossus altivelis). Bioinformatics analysis showed the structure of NOD2 to be highly conserved during vertebrate evolution. Dual-luciferase reporter assays examined the activation of NF-κB signaling and Western blotting analysis detected the phosphorylation of three MAP kinases (p-38, Erk1/2, and JNK1/2). Functional study revealed that, like its mammalian counterparts, PaNOD2 was the receptor of the bacterial cell wall component muramyl dipeptide (MDP), and the leucine-rich repeat motif was responsible for the recognition and binding of PaNOD2 with the ligand. Overexpression of PaNOD2 activated the NF-κB signaling pathway, leading to the upregulation of inflammatory cytokines, including TNF-α and IL-1ß in HEK293T cells and ayu head kidney-derived monocytes/macrophages (MO/MΦ). Particularly, we found that PaNOD2 activated the MAPK signaling pathways, as indicated by the increased phosphorylation of p-38, Erk1/2, and JNK1/2, which have not been characterized in any teleost species previously. Our findings proved that the NOD2 molecule and initiated pathways are conserved between mammals and ayu. Therefore, ayu could be used as an animal model to investigate NOD2-based diseases and therapeutic applications.

Rituximab (anti-CD20)-modified AZD-2014-encapsulated nanoparticles killing of B lymphoma cells.

The mTOR signal pathway is often highly activated in B-cell non-Hodgkin's lymphoma (NHL) and promotes cancer progression and chemo-resistance. Therefore, the pathways of mTOR are an important target for drug development in this disease. In the current study, we developed a rituximab (anti-CD20)-modified mTOR inhibitor, AZD-2014, loaded into nanoparticles (Ab-NPs-AZD-2014) for trial of its anti-NHL effect. In a cultured NHL cell line, Ab-NPs-AZD-2014 inhibited cancer cell growth, induced cell apoptosis, and blocked activation of mTORC1 and mTORC2 in Raji cells. These results indicate that antibody modification and nanomaterial loading of AZD-2014 with anti-CD20 significantly improved efficacy of AZD-2014 against NHL cells. This approach may ultimately deserve testing in therapy against NHL.

Toll-Like Receptors, Associated Biological Roles, and Signaling Networks in Non-Mammals.

The innate immune system is the first line of defense against pathogens, which is initiated by the recognition of pathogen-associated molecular patterns (PAMPs) and endogenous damage-associated molecular patterns (DAMPs) by pattern recognition receptors (PRRs). Among all the PRRs identified, the toll-like receptors (TLRs) are the most ancient class, with the most extensive spectrum of pathogen recognition. Since the first discovery of Toll in Drosophila melanogaster, numerous TLRs have been identified across a wide range of invertebrate and vertebrate species. It seems that TLRs, the signaling pathways that they initiate, or related adaptor proteins are essentially conserved in a wide variety of organisms, from Porifera to mammals. Molecular structure analysis indicates that most TLR homologs share similar domain patterns and that some vital participants of TLR signaling co-evolved with TLRs themselves. However, functional specification and emergence of new signaling pathways, as well as adaptors, did occur during evolution. In addition, ambiguities and gaps in knowledge still exist regarding the TLR network, especially in lower organisms. Hence, a systematic review from the comparative angle regarding this tremendous signaling system and the scenario of evolutionary pattern across Animalia is needed. In the current review, we present overview and possible evolutionary patterns of TLRs in non-mammals, hoping that this will provide clues for further investigations in this field.

Anti-GPC3 antibody-modified sorafenib-loaded nanoparticles significantly inhibited HepG2 hepatocellular carcinoma.

Sorafenib (SFB) has improved the treatment of hepatocellular carcinoma (HCC) and has fewer severe side effects than other agents used for that purpose. However, due to a lack of tumor-specific targeting, the concentration of the drug in tumor tissue cannot be permanently maintained at a level that inhibits tumor growth. To overcome this problem, we developed a novel SFB-loaded polymer nanoparticle (NP). The NP (a TPGS-b-PCL copolymer that was synthesized from ε-caprolactone and d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) via ring-opening polymerization) contains Pluronic P123 and SFB, and its surface is modified with anti-GPC3 antibody to produce the polymer nanoparticle (NP-SFB-Ab). The Ab-conjugated NPs had higher cellular uptake by HepG2 cells than did non-antibody-conjugated SPD-containing nanoparticles (NP-SFB). The NP-SFB-Ab also displayed better stability characteristics, released higher levels of SFB into cell culture medium, and was more cytotoxic to tumor cells than was non-targeted NP-SFB and free SFB. The NP-SFB-Ab downregulated expression of the anti-apoptosis molecule MCL-1, which led to polymerization of Bax and Bak in mitochondrial cytosol. The NP-SFB-AB also promoted the mitochondrial release of cytochrome C, resulting in cellular apoptosis. Moreover, the NP-SFB-Ab significantly inhibited the growth of HepG2 xenograft tumors in nude mice without producing obvious side effects. These findings suggest that NP-SFB-Ab is a promising new method for achieving targeted therapy of HCC.