Proteomics Identifies LUC7L3 as a Prognostic Biomarker for Hepatocellular Carcinoma.

Alternative splicing has been shown to participate in tumor progression, including hepatocellular carcinoma. The poor prognosis of patients with HCC calls for molecular classification and biomarker identification to facilitate precision medicine. We performed ssGSEA analysis to quantify the pathway activity of RNA splicing in three HCC cohorts. Kaplan-Meier and Cox methods were used for survival analysis. GO and GSEA were performed to analyze pathway enrichment. We confirmed that RNA splicing is significantly correlated with prognosis, and identified an alternative splicing-associated protein LUC7L3 as a potential HCC prognostic biomarker. Further bioinformatics analysis revealed that high LUC7L3 expression indicated a more progressive HCC subtype and worse clinical features. Cell proliferation-related pathways were enriched in HCC patients with high LUC7L3 expression. Consistently, we proved that LUC7L3 knockdown could significantly inhibit cell proliferation and suppress the activation of associated signaling pathways in vitro. In this research, the relevance between RNA splicing and HCC patient prognosis was outlined. Our newly identified biomarker LUC7L3 could provide stratification for patient survival and recurrence risk, facilitating early medical intervention before recurrence or disease progression.

People Have Systematically Different Ownership Intuitions in Seemingly Simple Cases.

Our understanding of ownership influences how we interact with objects and with each other. Here, we studied people's intuitions about ownership transfer using a set of simple, parametrically varied events. We found that people (N = 120 U.S. adults) had similar intuitions about ownership for some events but sharply opposing intuitions for others (Experiment 1). People (N = 120 U.S. adults) were unaware of these conflicts and overestimated ownership consensus (Experiment 2). Moreover, differences in people's ownership intuitions predicted their intuitions about the acceptability of using, altering, controlling, and destroying the owned object (N = 130 U.S. adults; Experiment 3), even when ownership was not explicitly mentioned (N = 130 U.S. adults; Experiment 4). Subject-level analyses suggest that these disagreements reflect at least two underlying intuitive theories, one in which intentions are central to ownership and another in which physical possession is prioritized.

Precision mapping child undernutrition for nearly 600,000 inhabited census villages in India.

There are emerging opportunities to assess health indicators at truly small areas with increasing availability of data geocoded to micro geographic units and advanced modeling techniques. The utility of such fine-grained data can be fully leveraged if linked to local governance units that are accountable for implementation of programs and interventions. We used data from the 2011 Indian Census for village-level demographic and amenities features and the 2016 Indian Demographic and Health Survey in a bias-corrected semisupervised regression framework to predict child anthropometric failures for all villages in India. Of the total geographic variation in predicted child anthropometric failure estimates, 54.2 to 72.3% were attributed to the village level followed by 20.6 to 39.5% to the state level. The mean predicted stunting was 37.9% (SD: 10.1%; IQR: 31.2 to 44.7%), and substantial variation was found across villages ranging from less than 5% for 691 villages to over 70% in 453 villages. Estimates at the village level can potentially shift the paradigm of policy discussion in India by enabling more informed prioritization and precise targeting. The proposed methodology can be adapted and applied to diverse population health indicators, and in other contexts, to reveal spatial heterogeneity at a finer geographic scale and identify local areas with the greatest needs and with direct implications for actions to take place.

Qinling liquid ameliorates renal immune inflammatory damage via activating autophagy through AMPK/Stat3 pathway in uric acid nephropathy.

BACKGROUND: Excessive deposition of uric acid (UA) is one of the risk factors for kidney damage. Qinling liquid (QL) has a certain therapeutic effect on uric acid nephropathy (UAN), but its regulation mechanism is still unclear. METHODS: UAN rat models and UA induced rat renal tubular epithelial cells (NRK-52E) were constructed to evaluate the functional roles of QL. We firstly evaluated the kidney function and the degree of kidney damage in rats after QL treatment. Then, effects of QL on autophagy and NLRP3 inflammasome activation were assessed. Moreover, the regulation of QL in AMPK and Stat3 phosphorylation levels and the relationship among autophagy, AMPK/Stat3 pathway and NLRP3 inflammasomes were determined. RESULTS: QL could alleviate the inflammatory damage in UAN rats and promote the activation of autophagy. In addition, QL suppressed UA-induced activation of NLRP3 inflammasomes in rat renal tubular epithelial cells, which was partially reversed by autophagy inhibitor. Further, AMPK/Stat3 axis-mediated autophagy participated in the regulation of UA-induced NLRP3 inflammasome activation in NRK-52E cells. Finally, we confirmed that inhibiting AMPK/Stat3 pathway partly deteriorated the ameliorating effect of QL on renal immune inflammatory injury in UAN rats. CONCLUSION: Through in vivo and in vitro experiments, we found that QL promotes autophagy by activating the AMPK/Stat3 pathway, thereby improving renal immune inflammatory injury in UAN.

Satellite observed reversal in trends of spring phenology in the middle-high latitudes of the Northern Hemisphere during the global warming hiatus.

The start of the growing season (SOS) is essential to track the responses of vegetation to climate change. However, recent findings on whether the SOS in the middle-high latitudes of the Northern Hemisphere (NH) continued to advance or reversed during the global warming hiatus were not consistent. It is necessary to investigate the causes of this controversy and to examine the relationship between the SOS and preseason temperature trends. To this end, we first applied four widely used phenology extraction methods to derive the SOS from the GIMMS NDVI3g dataset and then used the ensemble empirical modal decomposition (EEMD) method to extract the nonlinear trends of the SOS and preseason temperature. Our results clarify, for the first time, that the limitations of the linear assumption-based trend analysis methods are an important but overlooked cause of the discrepancies among existing studies on whether the SOS was advanced or delayed in the NH (>30° N) during the global warming hiatus. We further revealed the range of the mismatches between the SOS and preseason temperature trends at the latitude, altitude and biome levels. Specifically, we discovered that the SOS in the NH (>30° N) obtained by the four phenology extraction methods showed a significant reversal from advance to delay during the global warming hiatus, and the corresponding average rate of change was very small. The area showing increasing preseason temperatures decreased during the global warming hiatus, but it always occupied most of the NH (>30° N). However, delayed SOS trends were dominant in the NH from 50° N to 60° N, above 3000 m and in biomes other than TBMF and BF. Accordingly, using an EEMD-like approach to evaluate the changes in the SOS and preseason temperature is necessary for improving our understanding of the changes in the SOS and their association with climate.

The genetic alterations of rectal neuroendocrine tumor and indications for therapy and prognosis: a systematic review.

Neuroendocrine tumors (NETs) are a type of rare tumor that can occur at multiple organs. Rectal NETs are the most common NETs in gastrointestinal tract. Due to the rarity of rectal NETs in rectal cancer, the molecular features and the correlation with patient therapeutic response and prognosis have not been investigated in detail. In this review, we focused on the molecular features, potential therapeutic targets and prognosis of rectal NETs. By summarizing the relevant studies, we established the mutational landscape of rectal NETs and identified a series of large fragment variations. Driver genes including TP53, APC, KRAS, BRAF, RB1, CDKN2A and PTEN were found as the top mutated genes. Large fragment alterations mainly involved known driver genes, including APC, TP53, CCNE1, MYC, TERT, RB1 and ATM. Germline mutations of APC, MUTYH, MSH6, MLH1 and MSH2 associated with Lynch syndrome or FAP were also found in rectal NETs. The BRAF-V600E mutation was reported as an actionable target in rectal NETs, and the combined BRAF/MEK inhibitors were found to be effective targeting BRAF-V600E in advanced or metastatic NETs. The known prognostic risk factors of rectal adenocarcinoma, including a series of demographic and clinicopathological factors were also prognostic factors for rectal NETs. Furthermore, three types of markers, including genetic alterations, protein expression levels and methylation, were also suggested as prognostic factors for rectal NETs. In summary, we established the landscape of mutations and large-fragment alterations of rectal NETs, and identified potential therapeutic targets and a series of prognostic factors. Future studies may focus on the optimization of therapeutic strategies based on potential actionable biomarkers.

Prevalence and a correlation analysis of discoid meniscus and femoral trochlear dysplasia.

BACKGROUND: Discoid meniscus (DM) and femoral trochlear dysplasia (FTD) are common knee disorders. Both as congenital malformation, whether there is a connection between them is unclear and the research on their prevalence in the general population is inadequate. This study aimed to investigate the prevalence of FTD and DM in the general population through a large sample size, and to explore the relationship between them. STUDY DESIGN: Retrospective study. METHODS: Patients undergoing knee magnetic resonance imaging (MRI) examinations at our outpatient clinic were screened and 1003 patients were enrolled in DM group with 989 patients in non-DM (NDM) group. The type of DM and FTD was classified with Watanabe classification and Dejour's classification, respectively. The prevalence of FTD and DM in the general population and the relationship between them were evaluated. RESULTS: The prevalence of DM and FTD was 10.0% and 14.5%, respectively. The overall percentage of FTD was higher in DM group (P < 0.001). The DM group has a higher percentage of all types of FTD except type D (P < 0.05), and a higher percentage of both low- and high-grade FTD (P < 0.001). There were 633 cases of type I DM and 370 cases of type II DM. The overall percentage of FTD was not significantly different between the two types (P = 0.106). No significant difference was detected for all types of FTD except type B (P < 0.05). The Type I DM group has a significant higher percentage of high-grade FTD than Type II group (P < 0.05). CONCLUSION: Patients with a DM are more likely to have FTD regardless of the type of DM, while those with a type I DM are more prone to have a high grade FTD.

Preoperative serum calcium could be a prognostic factor for surgical treatment of recurrent patellar dislocation: a retrospective study.

BACKGROUND: Surgical treatment for recurrent patellar dislocation (RPD) could yield good outcomes. While, unsatisfactory recovery still exists in some cases. For all prognostic factors, serum biomarkers have rarely been investigated. This study aimed to evaluate the prognostic value of preoperative serum calcium level, a widely used serum biomarker, in surgical treatment for RPD. STUDY DESIGN: Retrospective study. METHODS: Ninety-nine patients with RPD were enrolled in the study. Preoperative serum calcium was acquired from routinely tested blood 1 day prior to operation. Demographic data, characteristics of RPD, postoperative functional outcomes were obtained. The association between preoperative calcium and postoperative functional outcomes (Kujala, Lysholm, Tegner, IKDC and KOOS score) was determined by correlation analysis and multivariate linear regression analysis. Poor recovery was determined as Kujala score below 80. The receiver operating characteristic (ROC) curve was used to assess the prognostic value of preoperative calcium. RESULTS: Patients were followed up for a mean period of 2.45 ± 1.33 years. All clinical scores showed significant improvement at the latest follow-up. Correlation and multivariate linear analyses indicated that serum calcium level was an important factor related with the prognosis of surgical treatment for RPD. According to the ROC curve, the cut-off value for preoperative calcium was 2.225 mmol/L. The clinical outcomes of patients with a preoperative blood calcium < 2.225 mmol/L was significantly worse than that with a higher calcium level. The correspondent sensitivity was 0.812 with a specificity of 0.633. CONCLUSION: Operative treatment for RPD achieved good results, while in some cases the functional scores remain inferior. As a serum biomarker, preoperative calcium could be prognostic for outcomes after surgical treatment for RPD.

Double-Tunnel Technique Was Similar to Single-Tunnel Technique in Clinical, Imaging and Functional Outcomes for Medial Patellofemoral Ligament Reconstruction: A Randomized Clinical Trial.

PURPOSE: This study aimed to compare the clinical, functional, and imaging outcomes of single-tunnel (ST) and double-tunnel (DT) techniques for medial patellofemoral ligament (MPFL) reconstruction. METHODS: Ninety-four patients with recurrent patellar instability were randomly divided into 2 groups, receiving either ST or DT MPFL reconstruction. Lateral reticulum release (LRR) and tibial tuberosity (TT) transfer were performed as combined procedures when necessary. Preoperative and postoperative clinical characteristics (symptoms and episodes of redislocation), functional outcomes (Kujala, Lysholm, Tegner, IKDC, and KOOS score), and radiological measurements (congruence angle, patellar tilt angle, lateral patellar angle, and lateral patellar translation) were analyzed. RESULTS: The analysis included data from 90 patients with 48 patients in the ST group and 42 patients in the DT group. Patients were followed up for a mean period of 37.8 (range: 27-50) months in the ST group and 38.6 (range: 25-53) months in the DT group. Forty-three patients in the ST group and 40 patients in the DT group received combined TT transfer, and all patients underwent LRR. At the latest follow-up, 1 patient in ST group experienced redislocation, while no patient in the DT group sustained clinical failure (P = .347). Imaging measurements decreased significantly to the normal range postoperatively. No significant difference was noted between the postoperative radiological results of the 2 groups. All clinical scores significantly improved postoperatively, and no significant difference was observed between the 2 groups except for the higher Lysholm score (P = .031), KOOS symptoms score (P = .021) and KOOS knee-related quality of life score (P = .043) in the DT group. CONCLUSION: Both techniques could equally mitigate the patellar lateral translation or redislocation. Our results demonstrate several significant differences in functional outcomes that favored DT MPFL reconstruction but no difference in clinical failure rates and radiological results between ST and DT MPFL reconstruction. LEVEL OF EVIDENCE: Level I, randomized clinical trial.

Allograft contamination during suture preparation for anterior cruciate ligament reconstruction: an ex vivo study.

PURPOSE: Effects of suture preparation on graft contamination remain unknown in anterior cruciate ligament reconstruction (ACLR). This study aimed to evaluate the incidence of allograft contamination at different time points of graft preparation and investigate differences in contamination between different sites of the allografts. METHODS: Fourteen hamstring tendon (HT), 9 quadriceps tendon (QT), and 9 bone-patellar tendon-bone (BTB) allografts were harvested, sterilised, and stored following routine procedures. Graft suture preparation was performed with baseball stitching for soft tissue and bone drilling for bone plug. The time was recorded simultaneously. The graft was kept moist in a standard operating room environment for 30 min after the initiation of preparation. The specimens were obtained from the middle and both ends of each graft for culture at three different time points: pre-suturing, post-suturing, and 30 min after the initiation of preparation. A total of 192 specimens were transferred to the microbiology laboratory for culture, identification, and semi-quantitative assessment. Culture results were classified as negative, poor, and abundant based on the extent of growth. Contamination level was recorded as low or high corresponding to culture results of poor or abundant. RESULTS: The duration of suture preparation was 348, 301, and 246 s for HT, QT, and BTB (P = 0.090). The specimens had a positive culture rate of 41/192 (21.4%), of which 21 were from the ends and 20 from the middle. More positive samples with abundant bacterial growth were detected from the ends than from the middles post-suturing (7/8 vs. 1/7, P = 0.010) and at 30 min (6/11 vs. 0/11, P = 0.012). The total graft contamination rate was significantly higher at 30 min (19/32, 59.4%) than pre-suturing (4/32, 15.6%) and post-suturing (9/32, 28.1%) (P < 0.001). The contamination rate with abundant bacterial growth was higher post-suturing (7/32, 21.9%) than pre-suturing (0%). No statistically significant differences were found among the three types of allografts. CONCLUSION: The contamination rate increases significantly at 30 min compared with pre-suturing and post-suturing. Suture preparation may have introduced the high-level contamination, to which the ends of the graft were more prone than the middle. Therefore, routine prophylactic decontamination after suture preparation should be considered, especially for the ends of the grafts.

The role of acetylation sites in the regulation of p53 activity.

As a "genomic guardian", p53 mainly functions as a transcription factor that regulates downstream targets responsible for cell fate control, and the activity of p53 is tightly regulated by a complex network that include an abundance of post-translational modifications. Notably, acetylation of p53 at many positions has been demonstrated to play a major role in accurate p53 regulation and cell fate determination. However, no evidence has been provided to compare the effect of acetylation at different sites on p53 regulation. Here, we constructed six acetylation-defective p53 mutants that lysine was substituted by arginine at residues 120, 164, 305, 320, 370/372/373 or 381/382/386, respectively, and determined their effects on p53 activity systematically. Our results showed that all six mutants exhibited diminished transactivation ability and selective regulation of target genes expression through distinct mechanisms. Specifically, lysine 370/372/373 and 381/382/386 mutations decreased p53 stability, and lysine 305 mutation reduced p53 phosphorylation level at serine 15, while lysine 120 and 164 mutations decreased p53 acetylation level at lysine 382. Collectively, these data indicate that acetylation of p53 at different sites has diverse regulatory effects on p53 transcriptional activity through different mechanisms.

Production and characterization of a monoclonal antibody for Pefloxacin and mechanism study of antibody recognition.

In this report, an artificial antigen (PFLX-BSA: Pefloxacin connected bovine serum albumin) was successfully prepared. The monoclonal antibody against pefloxacin was produced and characterized using a direct competitive ELISA. The linear range of detection was 0.115-6.564 µg/L. The limit of detection defined as IC15 was 0.170 ± 0.05 µg/L and the IC50 was 0.902 ± 0.03 µg/L. The antibody variable region genes were amplified, assembled, and sequenced. A three-dimensional structural model of the variable region was constructed to study the mechanism of antibody recognition using molecular docking analysis. Three predicted essential amino acids, Thr53, Arg97 of heavy chain and Thr52 of light chain, were mutated to verify the theoretical model. Three mutants lost binding activity significantly against pefloxacin as predicted. These may provide useful insights for studying antigen-antibody interaction mechanisms to improve antibody affinity maturation in vitro.

Enhanced biodegradation of atrazine by Arthrobacter sp. DNS10 during co-culture with a phosphorus solubilizing bacteria: Enterobacter sp. P1.

The interaction between pure culture microorganisms has been evaluated allowing for the enhanced biodegradation of various kinds of pollutants. Arthrobacter sp. DNS10 previously enriched in an atrazine-containing soil was capable of utilizing atrazine as the sole nitrogen source for growth, and Enterobacter sp. P1 is a phosphorus-solubilizing bacterium that releases various kinds of organic acids but lacks the ability to degrade atrazine. Whether strain P1 could enhance atrazine biodegradation by the degrader strain DNS10 was investigated in this experiment. Gas chromatography and high-performance liquid chromatography results showed that co-culture of both strains degraded 99.18 ±â€¯1.00% of the atrazine (initial concentration was 100 mg L-1), while the single strain DNS10 only degraded 38.57 ±â€¯7.39% after a 48 h culture, and the resulting concentration of the atrazine final metabolite cyanuric acid were 63.91 ±â€¯3.34 mg L-1 and 26.60 ±â€¯3.87 mg L-1, respectively. In addition, the expression of the atrazine degradation-related genes trzN, atzB and atzC in co-culture treatments was 6.61, 1.81 and 3.09 times that of the single strain DNS10 culture treatment. A substrates utilization test showed that the atrazine-degrading metabolites ethylamine and isopropylamine could serve as the nitrogen source to support strain P1 growth, although strain P1 cannot degrade atrazine or utilize atrazine for growth. Furthermore, the pH of the medium was significantly decreased when strain P1 utilized ethylamine and isopropylamine as the nitrogen source for growth. The results suggest that nondegrader strain P1 could promote the atrazine biodegradation when co-cultured with strain DNS10. This phenomenon is due to metabolite exchange between the two strains. Culturing these two strains together is a new biostimulation strategy to enhance the biodegradation of atrazine by culturing these two strains together.

Colorimetric bio-barcode immunoassay for parathion based on amplification by using platinum nanoparticles acting as a nanozyme.

A competitive bio-barcode immunoassay is described for the trace detection of parathion in water, pear, cabbage, and rice samples. It is based on amplification by platinum nanoparticle acting as a nanozyme. Gold nanoparticles (AuNPs) were modified with (a) monoclonal antibodies (mAbs) against parathion, and (b) thiolated single-stranded DNA (ssDNA) oligonucleotides. Magnetic nanoparticles (MNPs) were functionalized with ovalbumin coupled with parathion hapten. Parathion and its hapten compete with mAbs on the surface of the AuNPs. Subsequently, the platinum nanoparticles (PtNPs) probe, which was functionalized with the complementary thiolated ssDNA (C-ssDNA), was added to the reaction mixture for the detection of parathion. The signal was catalytically amplified by coupling with platinum nanozyme using teramethylbenzidine and H2O2 as the chromogenic system. The immunoassay has a linear range that extends from 0.01-50 µg·L-1, and the limit of detection is 2.0 × 10-3 µg·L-1. The recoveries and relative standard deviations (RSDs) ranged from 91.1-114.4% and 3.6-15.8%, respectively. The method correlates well with data obtained by gas chromatography-tandem mass spectrometry (GC-MS/MS). Graphical abstract The parathion and the magnetic nanoparticles (MNPs) labelled with hapten-OVA competitively reacted to AuNPs modified with mAbs and thiolated DNA for the detection of parathion. The signal was catalyzed by platinum nanozyme. The limit of detection for parathion is 2.0 ng·L-1.

Sensitive detection of Campylobacter jejuni using one-step strategy based on functional nanospheres of immunomagnetic capture and quantum dots.

Campylobacter jejuni has emerged as the most common bacterial foodborne illness in the developed world. Here, we demonstrate a convenient one-step strategy for detecting C. jejuni. Immunomagnetic nanospheres (IMNS) and immunofluorescent nanospheres (IFNS, quantum dots) were used for the simultaneous, sensitive capture and recognition of C. jejuni. After magnetic separation with the IMNS, detection of C. jejuni was achieved with fluorescence measurement of the IFNS in the sandwich complexes (IMNS-bacteria-IFNS). The limit of detection of this assay was 103 CFU/mL, and the linear range was from 105 to 107 CFU/mL (R2 = 0.9994). When compared with a conventional two-step detection strategy, in which C. jejuni was first captured with the IMNS and then detected using the IFNS, this one-step detection strategy enhance sensitivity and save time. This suggested that the developed method has the potential for use as an alternative to the standard method for food quality assurance, as it provides rapid detection of C. jejuni in foodstuffs and the environment.

Inflammasome-Derived Exosomes Activate NF-κB Signaling in Macrophages.

Exosomes are secreted small vesicles that mediate various biological processes, such as tumorigenesis and immune response. However, whether the inflammasome signaling leads to the change of constituent of exosomes and its roles in immune response remains to be determined. We isolated the exosomes from macrophages with treatment of mock, endotoxin, or endotoxin/nigericin. A label-free quantification method by MS/MS was used to identify the components of exosomes. In total, 2331 proteins were identified and 513 proteins were exclusively detected in exosomes with endotoxin and nigericin treatment. The differentially expressed proteins were classified by Gene Ontology and KEGG pathways. The immune response-related proteins and signaling pathways were specifically enriched in inflammasome-derived exosomes. Moreover, we treated macrophages with the exosomes from different stimulation. We found that inflammasome-derived exosomes directly activate NF-κB signaling pathway, while the control or endotoxin-derived exosomes have no effect. The inflammatory signaling was amplified in neighbor cells in an exosome-dependent way. The inflammasome-derived exosomes might be used to augment the immune response in disease treatment, and preventing the transfer of these exosomes might ameliorate autoimmune diseases.

A Light Responsive Nanoparticle-Based Delivery System Using Pheophorbide A Graft Polyethylenimine for Dendritic Cell-Based Cancer Immunotherapy.

In this study, the photochemical internalization (PCI) technique was adopted in a nanoparticle-based antigen delivery system to enhance antigen-specific CD8+ T cell immune response for cancer immunotherapy. Pheophorbide A, a hydrophobic photosensitizer, grafted with polyethylenimine (PheoA-PEI) with endosome escape activity and near-infrared imaging capability was prepared. A model antigen ovalbumin (OVA) was then complexed with PheoA-PEI to form PheoA-PEI/OVA nanoparticles (PheoA-PEI/OVA NPs) that are responsive to light. Flow cytometry analysis revealed increased endocytosis in a murine dendritic cell line (DC2.4) that was treated with PheoA-PEI/OVA NPs compared to free OVA. Generation of reactive oxygen species (ROS) in DC2.4 cells was also confirmed quantitatively and qualitatively using 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA). Confocal laser scanning microscopy (CLSM) further demonstrated that the PheoA-PEI/OVA NPs enhanced cytosolic antigen release after light stimulation. Moreover, PheoA-PEI/OVA NP treated DC2.4 cells exhibited enhanced cross-presentation to B3Z T cell hybridoma in vitro after light irradiation, substantially increased compared to those treated with free OVA. Consistently, in vivo results revealed upregulation of CD3+CD8+T lymphocytes in tumors of mice treated with dendritic cells plus PheoA-PEI/OVA NPs and light irradiation. The activated T cell response is partly responsible for the inhibitory effect on E.G7 tumor growth in mice immunized with dendritic cells plus PheoA-PEI/OVA NPs and light irradiation. Our results demonstrate the feasibility to enhance antigen-specific CD8+ T cell immune response by light-responsive nanoparticle-based vaccine delivery for cancer immunotherapy.

Variation in the Humification Degree of Dissolved Organic Matter from Cattle Manure during Composting as Analyzed by Ultraviolet-Visible and Fluorescence Spectroscopy.

This study investigated the degree of humification of dissolved organic matter (DOM) during different periods of cattle manure composting using ultraviolet-visible (UV-vis) and fluorescence spectroscopy (emission, synchronous scan, and excitation-emission matrix) and determined which method is more suitable for analysis of the humification degree of DOM. Two composting piles were prepared by mixing manure and corn straw. One pile (Pile A [PA]) contained inoculated exogenous composite agents at a ratio of 2% (v/v), and a pile without the addition of inoculants (PNA) served as the control treatment. The results showed that ultraviolet integrated absorption intensities in the range of 226 to 400 nm and 260 to 280 nm and specific ultraviolet absorbances at 254 and 280 nm of both PA and PNA gradually increased with composting time. Based on the fluorescence regional integration analysis and parallel factor analysis, the humic-like substances became the main components of the DOM after composting. Our study demonstrated that the humification degree of DOM was enhanced during composting and that the inoculation composite agent was beneficial for the humification of DOM at the mesophilic and thermophilic phases of the composting process. Moreover, the results of correlation analysis and principal component analysis demonstrated that the fluorescence spectral parameters evaluated the humification degree of DOM during the whole cattle manure composting process better than the UV-vis spectral parameters.

Cannabinoid 2 Receptor Agonist Improves Systemic Sensitivity to Insulin in High-Fat Diet/Streptozotocin-Induced Diabetic Mice.

BACKGROUND/AIMS: The endocannabinoid signalling (ECS) system has been known to regulate glucose homeostasis. Previous studies have suggested that the cannabinoid 2 (CB2) receptor may play a regulatory role on insulin secretion, immune modulation and insulin resistance. Given that diabetes and insulin resistance are attributable to elevated inflammatory tone, we investigated the role of CB2 receptor on glucose tolerance and insulin sensitivity in high-fat diet (HFD)/streptozotocin (STZ)-induced mice. METHODS: Diabetes was induced in male ICR mice by HFD/STZ and exposed to a CB2 receptor agonist, SER601, for 2- or 4-weeks via subcutaneous implantation of osmotic minipumps. Glucose and insulin tolerance tests were performed at the end of treatment. Islets were isolated for assessment of ß-cell function. Pancreases and skeletal muscles were also obtained for histological analyses. RESULTS: Despite a lack of impact on glucose tolerance, substantial improvement on insulin sensitivity was observed in SER601-treated mice, which could partly be attributed to improved islet ß-cell function, shown as increased glucose-induced insulin secretion and insulin content. No changes on islet macrophage infiltration or skeletal muscle fat deposition were detectable from SER601-treated mice. However, a major decrease in body weight was recorded at the end of 4-week SER601 exposure, accompanied by a lack of epididymal adipose mass in SER601-treated mice. CONCLUSION: Our data suggest a lipolytic role of SER601 in HFD/STZ-induced diabetic mice, which results in significant improvement of systemic insulin sensitivity. Thus, the CB2 receptor may be considered a promising target for therapeutic development against insulin resistance and obesity-related diabetes.

Production of a soluble single-chain variable fragment antibody against okadaic acid and exploration of its specific binding.

Okadaic acid is a lipophilic marine algal toxin commonly responsible for diarrhetic shellfish poisoning (DSP). Outbreaks of DSP have been increasing and are of worldwide public health concern; therefore, there is a growing demand for more rapid, reliable, and economical analytical methods for the detection of this toxin. In this study, anti-okadaic acid single-chain variable fragment (scFv) genes were prepared by cloning heavy and light chain genes from hybridoma cells, followed by fusion of the chains via a linker peptide. An scFv-pLIP6/GN recombinant plasmid was constructed and transformed into Escherichia coli for expression, and the target scFv was identified with IC-CLEIA (chemiluminescent enzyme immunoassay). The IC15 was 0.012 ± 0.02 µg/L, and the IC50 was 0.25 ± 0.03 µg/L. The three-dimensional structure of the scFv was simulated with computer modeling, and okadaic acid was docked to the scFv model to obtain a putative structure of the binding complex. Two predicted critical amino acids, Ser32 and Thr187, were then mutated to verify this theoretical model. Both mutants exhibited significant loss of binding activity. These results help us to understand this specific scFv-antigen binding mechanism and provide guidance for affinity maturation of the antibody in vitro. The high-affinity scFv developed here also has potential for okadaic acid toxin detection.