Wheat straw biochar as an additive in swine manure Composting: An in-depth analysis of mixed material particle characteristics and interface interactions.

In recent research, biochar has been proven to reduce the greenhouse gases and promote organic matter during the composting. However, gas degradation may be related to the microstructure of compost. To investigate the mechanism of biochar additive, composting was performed using swine manure, wheat straw and biochar and representative solid compost samples were analyzed to characterize the mixed biochar and compost particles. We focused on the microscale, such as the particle size distributions, surface morphologies, aerobic layer thicknesses and the functional groups. The biochar and compost particle agglomerations gradually became weaker and the predominant particle size in the experiment group was < 200 µm. The aerobic layer thickness (Lp) was determined by infrared spectroscopy using the wavenumbers 2856 and 1568 cm-1, which was 0-50 µm increased as composting proceeded in both groups. The biochar increased Lp and facilitated oxygen penetrating the compost particle cores. Besides, in the biochar-swine manure particle interface, the aliphatic compound in the organic components degraded and the content of aromaticity increased with the composting process, which was indicated by the absorption intensity at 2856 cm-1 decreasing trend and the absorption intensity at 1568 cm-1 increasing trend. In summary, biochar performed well in the microscale of compost pile.

Large Semi-Membrane Covered Composting System Improves the Spatial Homogeneity and Efficiency of Fermentation.

Homogenous spatial distribution of fermentation characteristics, local anaerobic conditions, and large amounts of greenhouse gas (GHGs) emissions are common problems in large-scale aerobic composting systems. The aim of this study was to examine the effects of a semi-membrane covering on the spatial homogeneity and efficiency of fermentation in aerobic composting systems. In the covered group, the pile was covered with a semi-membrane, while in the non-covered group (control group), the pile was uncovered. The covered group entered the high-temperature period earlier and the spatial gradient difference in the group was smaller compared with the non-covered group. The moisture content loss ratio (5.91%) in the covered group was slower than that in the non-covered group (10.78%), and the covered group had a more homogeneous spatial distribution of water. The degradation rate of organic matter in the non-covered group (11.39%) was faster than that in the covered group (10.21%). The final germination index in the covered group (85.82%) was higher than that of the non-covered group (82.79%) and the spatial gradient difference in the covered group was smaller. Compared with the non-covered group, the oxygen consumption rate in the covered group was higher. The GHG emissions (by 30.36%) and power consumption in the covered group were reduced more significantly. The spatial microbial diversity of the non-covered group was greater compared with the covered group. This work shows that aerobic compost covered with a semi-membrane can improve the space homogeneity and efficiency of fermentation.

A novel in-situ quantitative profiling approach for visualizing changes in lignin and cellulose by stained micrographs.

There is a strong need for low-cost lignocellulosic composition simultaneous localization methodologies to benefit deeper understandings of crop stalk morphology. This study developed a robust quantitative safranin O-fast green staining-based optical microscopy imaging methodology for in-situ simultaneously generating digital profiles of lignin and cellulose in stalk tissues. Foreground extraction and dye residue removal of stained images were adapted. The ratios of normalized red (R), green (G), and blue (B) channel signal intensity, R/B and G/B, were defined as quantitative indicators of lignin and cellulose, respectively. The method was validated on model rice with known bioinformatics, and the results were consistent with those of fluorescence microscopy and immunogold labeling methods. The high-definition spatial in-situ simultaneous profiles of lignin and cellulose in alkali-treated maize stalk tissues and their variations were visualized. This low-cost, cell-scale method is expected to contribute to new discoveries in many areas of biomass refining and plant science.

Natural variation of DROT1 confers drought adaptation in upland rice.

Upland rice is a distinct ecotype that grows in aerobic environments and tolerates drought stress. However, the genetic basis of its drought resistance is unclear. Here, using an integrative approach combining a genome-wide association study with analyses of introgression lines and transcriptomic profiles, we identify a gene, DROUGHT1 (DROT1), encoding a COBRA-like protein that confers drought resistance in rice. DROT1 is specifically expressed in vascular bundles and is directly repressed by ERF3 and activated by ERF71, both drought-responsive transcription factors. DROT1 improves drought resistance by adjusting cell wall structure by increasing cellulose content and maintaining cellulose crystallinity. A C-to-T single-nucleotide variation in the promoter increases DROT1 expression and drought resistance in upland rice. The potential elite haplotype of DROT1 in upland rice could originate in wild rice (O. rufipogon) and may be beneficial for breeding upland rice varieties.

New rapid detection method of total chlorogenic acids in plants using SERS based on reusable Cu2O-Ag substrate.

A new method for rapidly detecting of total chlorogenic acids (CGAs) in plants by surface-enhanced Raman spectroscopy (SERS) based on reusable Cu2O-Ag substrate was developed in this study. The Cu2O-Ag substrate prepared by the in-situ growth method had high uniformity with peak intensity relative standard deviation (RSD) of 5.27%, repeatability with peak intensity RSD of 3.58%, and sensitivity with an analytical enhancement factor of 1.27 × 105 for detecting CGAs. Furthermore, the substrate had excellent reusability, after it was reused for seven cycles, the signal strength of CGAs was still above 80% of initial. Compared with the standard method of high-performance liquid chromatography (HPLC), the SERS method can successfully analyze the contents of total CGAs in plants, such as Stevia rebaudiana leaves, coffee beans, Lonicera japonica leaves, and Eucommia ulmoides flowers, with recovery rate from 93.26% to 112.65%, and the limit of detection was 0.13 µg/mL. The total CGAs content of Stevia rebaudiana leaves samples detected by HPLC and SERS have good consistency with R = 0.9760 and RMSE = 3286 mg/kg. Furthermore, the SERS method only needed less than 1 min, one standard and reusable substrate in this study to analyze, which can further reduce the cost of method analysis. Therefore, the SERS method with the appropriate substrate can provide a rapid, accurate, and economic way to detect the total CGAs in plants.

Rapid and simultaneous detection of multiple illegal additives in feed and food by SERS with reusable Cu2O-Ag/AF-C3N4 substrate.

Illegal additives can bring the economic benefit, resulting in the continuous irregularities in the use of illegal additives. In this study, a method for rapid, sensitive, and simultaneous detection of multiple illegal additives including enrofloxacin, malachite green, nitrofurazone, and Sudan Ⅰ in feed and food samples by surface-enhanced Raman spectroscopy (SERS) with Cu2O-Ag/AF-C3N4 composite substrate was developed. A Cu2O-Ag/AF-C3N4 composite substrate was prepared by reacting Cu2O modified by AF-C3N4 nanosheets with AgNO3 solution. The substrate has a limit of detection (LOD) of 1.29 × 10-6 mg/L, a good linear relationship of between 10-6 and 10-2 mg/L, and an R2 value of 0.95 for Rhodamine B detection. Furthermore, the substrate showed high uniformity and reproducibility, with relative standard deviations (RSD) of 6.74% and 4.85%, respectively. Adding AF-C3N4 nanosheets not only increased the enhancement effect of the substrate, which was 4.4 times of that before addition, but also endowed it with good self-cleaning characteristics owing to its excellent photocatalytic activity. The substrate can be reused, with over 80% of the original Raman signal strength remaining after four repeat uses. The SERS based on the above substrate was used to detect the illegal additives, the LOD of enrofloxacin, malachite green, nitrofurazone, and Sudan Ⅰ can reach 4.67 × 10-4 mg/L, 2.57 × 10-5 mg/L, 5.7 × 10-7 mg/L and 6.92 × 10-5 mg/L. The results reveal that this substrate has great application potential in feed and food safety.

Chronic obstructive pulmonary disease is characterized by reduced levels and defective suppressive function of regulatory B cells in peripheral blood.

Chronic obstructive pulmonary disease (COPD) is characterized by a progressive, persistent immune response to cigarette smoke, and it has been suggested that immune dysregulation is involved in its pathogenesis. A subset of regulatory B cells (Bregs) with high levels of the surface markers CD24 and CD38 (CD24hiCD38hi) has previously been shown to exert an immunosuppressive function. This study investigated the levels and activity of CD24hiCD38hi Bregs in stable COPD (sCOPD). Testing the peripheral blood from 65 patients with sCOPD and 39 control subjects for CD24hiCD38hi Breg subsets by flow cytometry showed that the patients with sCOPD had significantly lower levels of CD24hiCD38hi Bregs and IL-10+ B cells. The patients with sCOPD had lower serum interleukin-10 levels than the controls. The patients with most severe sCOPD had the lowest levels of CD24hiCD38hi Bregs. Spearman correlation analysis showed that the levels of CD24hiCD38hi Bregs in the patients with sCOPD positively correlated with serum interleukin-10 concentrations but not with levels of C-reactive protein. Compared to healthy controls, functional studies showed that Breg cells from patients with sCOPD exhibit a decreased suppressive function. We conclude that sCOPD is characterized by the exhaustion of CD24hiCD38hi regulatory B cells compartment. Therefore, CD24hiCD38hi Bregs may contribute to the pathogenesis of sCOPD.

A novel analytical strategy for discriminating antibiotic mycelial residue adulteration in feed based on ATR-IR and microscopic infrared imaging.

The Antibiotic mycelial residue (AMR) contains antibiotic residue, there are safety risks if it is used illegally in feed. This study investigated the feasibility of qualitative identification of AMR in protein feed and self-prepared feed based on attenuated total reflection mid-infrared spectrum (ATR-IR) and microscopic infrared imaging. Cottonseed meal (CM), soybean meal (SM), distillers dried grains with solubles (DDGS), nucleotide residue (NR), oxytetracycline residue (OR) and streptomycin sulfate residue (SR) and two self-prepared feed (broiler and pig) were used as research objects. The results showed that there were characteristic peaks at 1614 cm-1, 1315 cm-1, 779 cm-1, 514 cm-1 in the ATR-IR spectra of AMR, which were related to calcium oxalate hydrate. After detection, the content of total calcium and calcium oxalate in AMR were higher than those in protein feed. ATR-IR can quickly realize the qualitative discrimination of pure material samples. The combination of ATR-IR and partial least squares discriminant analysis (PLSDA) was effective in discriminating AMR from CM and SM with a single component (the classification errors were 0), but it cannot meet the discrimination of AMR from the fermented protein feed (such as DDGS and NR, the classification errors were 0.10 and 0.12) and self-prepared feed with complex components. Compared with ATR-IR, microscopic infrared imaging was less affected by the sample complexity. Multi-component samples belong to physical mixing and will not affect the infrared spectra of each component. Therefore, microscopic infrared imaging combined with effective information extraction algorithms such as cosine similarity can distinguish OR in the fermented protein feed and self-prepared feed. The above results showed that the advantages of ATR-IR and microscopic infrared imaging were complementary, which provided a new idea for the discrimination analysis of illegal feed additives.

[Technical evaluation and principle analysis of simulative habitat cultivation of Dendrobium nobile].

The technique of "simulative habitat cultivation" is to preserve the quality of traditional Chinese medicine by simulating the original habitat and site environment of wild Chinese medicine resources. Dendrobium nobile is the most representative variety of traditional Chinese medicine which reflects the coordinated development of medicinal material production and ecological environment. In this paper, the main technical points of the simulated cultivation model of D. nobile were summarized as follows: rapid propagation of seedling tissue technology to ensure the genetic stability of provenance; line card+fermented cow manure+live moss method to improve the survival rate; epiphytic stone cultivation to improve the quality of medicinal materials; and the integration of mycorrhizal fungi to improve the quality stability of medicinal materials. On the basis of summarizing the ecological benefits, economical and social benefits generated by the application of the technology, the paper systematically analyzes the principle of the technology for the cultivation of D. nobile to promote the excellent quality, the light, gas, heat and fertilizer resources of the undergrowth niche are in line with the wild site environment of D. nobile. The rich and complex soil microbial community in the forest laid the foundation for the species diversity needed for the growth of D. nobile.The stress effect on the growth of D. nobile resulted in the accumulation of secondary metabolites. The symbiotic relationship between the symbiotic fungi such as bryophytes and D. nobile promotes the synthesis of plant secondary metabolites. The high quality D. nobile was produced efficiently by improving and optimizing the cultivation techniques.

Application of nomograms to predict overall and cancer-specific survival in patients with chordoma.

BACKGROUND: The survival prediction of patients with chordoma is difficult to make due to the rarity of this oncologic disease. Our objective was to apply a nomogram to predict survival outcomes in individuals with chordoma of the skull base, vertebral column, and pelvis. METHODS: A total of 558 patients with chordoma between 1973 and 2014 were collected from the Surveillance, Epidemiology, and End Results (SEER) database. Independent prognostic factors in patients with chordoma were identified via univariate and multivariate Cox analysis. Then these prognostic factors were incorporated into a nomogram to predict 3- and 5-year overall survival and cancer-specific survival rates. Internal and external data were used to validate the nomograms. Concordance indices (C-indices) were used to estimate the accuracy of this nomogram system. RESULTS: A total of 558 patients were randomly assigned into a training cohort (n = 372) and a validation cohort (n = 186). Age, surgical stage, tumor size, histology, primary site, and use of surgery were identified as independent prognostic factors via univariate and multivariate Cox analysis (all p < 0.05) and further included to establish the nomogram. The C-indices for overall survival and cancer-specific survival prediction of the training cohort were 0.775 (95% confidence interval, 0.770-0.779) and 0.756 (95% confidence interval, 0.749 -0.762). The calibration plots both showed an excellent consistency between actual survival and nomogram prediction. CONCLUSION: Nomograms were constructed to predict overall survival and cancer-specific survival for patients with chordoma of the skull base, vertebral column, and pelvis. The nomogram could help surgeons to identify high risk of mortality and evaluate prognosis in patients with chordoma.

Diffusion Tensor Magnetic Resonance Imaging in Chronic Spinal Cord Compression.

Chronic spinal cord compression is the most common cause of spinal cord impairment in patients with nontraumatic spinal cord damage. Conventional magnetic resonance imaging (MRI) plays an important role in both confirming the diagnosis and evaluating the degree of compression. However, the anatomical detail provided by conventional MRI is not sufficient to accurately estimate neuronal damage and/or assess the possibility of neuronal recovery in chronic spinal cord compression patients. In contrast, diffusion tensor imaging (DTI) can provide quantitative results according to the detection of water molecule diffusion in tissues. In the present study, we develop a methodological framework to illustrate the application of DTI in chronic spinal cord compression disease. DTI fractional anisotropy (FA), apparent diffusion coefficients (ADCs), and eigenvector values are useful for visualizing microstructural pathological changes in the spinal cord. Decreased FA and increases in ADCs and eigenvector values were observed in chronic spinal cord compression patients compared to healthy controls. DTI could help surgeons understand spinal cord injury severity and provide important information regarding prognosis and neural functional recovery. In conclusion, this protocol provides a sensitive, detailed, and noninvasive tool to evaluate spinal cord compression.

Persistence and outcomes of non-vitamin K antagonist oral anticoagulants versus warfarin in patients with non-valvular atrial fibrillation.

AIMS AND OBJECTIVES: To compare persistence and outcomes of non-vitamin K antagonist oral anticoagulants (NOACs) versus warfarin in Chinese patients with non-valvular atrial fibrillation (AF). BACKGROUND: Given the unpredictable warfarin response and the costliness of NOACs, more research is needed to clarify which drug enjoys better persistence and outcomes, helping to provide personalised care for patients. DESIGN: A prospective cohort study. METHODS: Chinese patients taking NOACs or warfarin from March 2016-April 2018 were followed up by telephone or outpatient visit at 3, 6 months and half a year thereafter. Anticoagulant persistence and outcomes including stroke and bleeding were collected. We used Cox regression to analyse data. This study was reported according to the STROBE guideline. RESULTS: A total of 344 patients were enrolled; 146 patients received NOACs including dabigatran and rivaroxaban, and 198 patients received warfarin. Persistence with anticoagulants was low and dropped sharply at the third month. Patients on NOACs had worse persistence at 3, 6 and 12 months than those on warfarin. There was no difference in the incidence of ischaemic stroke and bleeding between groups, although ischaemic stroke and major bleeding occurred less frequently in the NOACs group. Paroxysmal AF, no heart failure and no stroke were predictors of NOACs non-persistence. Prior catheter ablation and no diabetes were associated with poor persistence of warfarin. The main reason for anticoagulant cessation was patient preference. CONCLUSIONS: Chinese patients taking NOACs had lower persistence, similar rate of ischaemic stroke and bleeding compared with those on warfarin. Further inventions are needed to improve persistence in Chinese patients on NOACs. RELEVANCE TO CLINICAL PRACTICE: Anticoagulation should highlight both persistence and outcomes emphasising personalised care of different drugs. Further interventions to improve persistence should be developed based on causes and risk factors and carried out in the third month of therapy.

Nomogram application to predict overall and cancer-specific survival in osteosarcoma.

PURPOSE: A prognostic nomogram was applied to predict survival in osteosarcoma patients. PATIENTS AND METHODS: Data collected from 2,195 osteosarcoma patients in the Surveillance, Epidemiology, and End Results (SEER) database between 1983 and 2014 were analyzed. Independent prognostic factors were identified via univariate and multivariate Cox analyses. These were incorporated into a nomogram to predict 3- and 5-year overall survival (OS) and cancer-specific survival (CSS) rates. Internal and external data were used for validation. Concordance indices (C-indices) were used to estimate nomogram accuracy. RESULTS: Patients were randomly assigned into a training cohort (n=1,098) or validation cohort (n=1,097). Age at diagnosis, tumor site, histology, tumor size, tumor stage, use of surgery, and tumor grade were identified as independent prognostic factors via univariate and multivariate Cox analyses (all P<0.05) and then included in the prognostic nomogram. C-indices for OS and CSS prediction in the training cohort were 0.763 (95% CI 0.761-0.764) and 0.764 (95% CI 0.762-0.765), respectively. C-indices for OS and CSS prediction in the external validation cohort were 0.739 (95% CI 0.737-0.740) and 0.740 (95% CI, 0.738-0.741), respectively. Calibration plots revealed excellent consistency between actual survival and nomogram prediction. CONCLUSION: Nomograms were constructed to predict OS and CSS for osteosarcoma patients in the SEER database. They provide accurate and individualized survival prediction.

Identification of microRNAs in acute respiratory distress syndrome based on microRNA expression profile in rats.

Acute respiratory distress syndrome (ARDS) remains a severe disease associated with an ~40% mortality rate and as many as 200,000 new cases annually. MicroRNAs (miRNAs) have important roles in gene regulation and cancer development. The present study aimed to identify the potential roles of miRNAs in the pathogenesis and progression of ARDS. The miRNA expression profile of the GSE57223 dataset was downloaded from the Gene Expression Omnibus database. Following data normalization, differentially expressed miRNAs were identified using the t­test method. The miRWalk database was searched to predict target genes of the identified miRNAs and then a miRNA­miRNA network with co­regulated target genes was constructed. Additionally, Gene Ontology (GO) analysis was performed for the target genes and a miRNA­miRNA functional synergistic network (MFSN) was established. GO and pathway analyses were performed for the co­regulated target genes of significant miRNAs in MFSN. Additionally, a protein­protein­interaction network was constructed for these target genes. A total of 19 miRNAs were differentially expressed between ARDS and normal lung tissue were identified. The four downregulated rno­let­7 family members were detected to have numerous co­regulated target genes and synergistic functions. Additionally, the target genes of the four miRNAs were significantly enriched the biological processes of wounding and inflammatory response. Additionally, interleukin (IL)­6 was identified as a hub protein with a high degree. The four downregulated rno­let­7 miRNAs may be involved in the inflammatory process in the pathogenesis and progression of ARDS, via the synergistic regulation of their target genes, such as IL­6. However, additional experimental validation is required.

Mir-17∼92 Governs Motor Neuron Subtype Survival by Mediating Nuclear PTEN.

Motor neurons (MNs) are unique because they project their axons outside of the CNS to innervate the peripheral muscles. Limb-innervating lateral motor column MNs (LMC-MNs) travel substantially to innervate distal limb mesenchyme. How LMC-MNs fine-tune the balance between survival and apoptosis while wiring the sensorimotor circuit en route remains unclear. Here, we show that the mir-17∼92 cluster is enriched in embryonic stem cell (ESC)-derived LMC-MNs and that conditional mir-17∼92 deletion in MNs results in the death of LMC-MNs in vitro and in vivo. mir-17∼92 overexpression rescues MNs from apoptosis, which occurs spontaneously during embryonic development. PTEN is a primary target of mir-17∼92 responsible for LMC-MN degeneration. Additionally, mir-17∼92 directly targets components of E3 ubiquitin ligases, affecting PTEN subcellular localization through monoubiquitination. This miRNA-mediated regulation modulates both target expression and target subcellular localization, providing LMC-MNs with an intricate defensive mechanism that controls their survival.

Fab fragment glycosylated IgG may play a central role in placental immune evasion.

STUDY QUESTION: How does the placenta protect the fetus from immune rejection by the mother? SUMMARY ANSWER: The placenta can produce IgG that is glycosylated at one of its Fab arms (asymmetric IgG; aIgG) which can interact with other antibodies and certain leukocytes to affect local immune reactions at the junction between the two genetically distinct entities. WHAT IS KNOWN ALREADY: The placenta can protect the semi-allogenic fetus from immune rejection by the immune potent mother. aIgG in serum is increased during pregnancy and returns to the normal range after giving birth. aIgG can react to antigens to form immune complexes which do not cause a subsequent immune effector reaction, including fixing complements, inducing cytotoxicity and phagocytosis, and therefore has been called 'blocking antibody'. STUDY DESIGN, SIZE, DURATION: Eighty-eight human placentas, four trophoblast cell lines (TEV-1, JAR, JEG and BeWo), primary culture of human placental trophoblasts and a gene knock-out mouse model were investigated in this study. PARTICIPANTS/MATERIALS, SETTING, METHODS: The general approach included the techniques of cell culture, immunohistochemistry, in situ hybridization, immuno-electron microscopy, western blot, quantitative PCR, protein isolation, glycosylation analysis, enzyme digestion, gene sequencing, mass spectrophotometry, laser-guided microdissection, enzyme-linked immunosorbent assay, pulse chase assay, double and multiple staining to analyze protein and DNA and RNA analysis at the cellular and molecular levels. MAIN RESULTS AND THE ROLE OF CHANCE: Three major discoveries were made: (i) placental trophoblasts and endothelial cells are capable of producing IgG, a significant portion of which is aberrantly glycosylated at one of its Fab arms to form aIgG; (ii) the asymmetrically glycosylated IgG produced by trophoblasts and endothelial cells can react to immunoglobulin molecules of human, rat, mouse, goat and rabbit at the Fc portion; (iii) asymmetrically glycosylated IgG can react to certain leukocytes in the membrane and cytoplasm, while symmetric IgG from the placenta does not have this property. LIMITATIONS, REASONS FOR CAUTION: Most of the experiments were performed in vitro. The proposed mechanism calls for verification in normal and abnormal pregnancy. WIDER IMPLICATIONS OF THE FINDINGS: This study identified a number of new phenomena suggesting that aIgG produced by the placenta would be able to react to detrimental antibodies and leukocytes and interfere with their immune reactions against the placenta and the fetus. This opens a new dimension for further studies on pregnancy physiology and immunology. Should the mechanism proposed here be confirmed, it will have a direct impact on our understanding of the physiology and pathology of human reproduction and offer new possibilities for the treatment of many diseases including spontaneous abortion, infertility and pre-eclampsia. It also sheds light on the mechanism of immune evasion in general including that of cancer.

Hyperbaric oxygen enlarges the area of brain damage in MCAO rats by blocking autophagy via ERK1/2 activation.

Hyperbaric oxygen (HBO) is emerging as a therapy for brain ischemia, although its benefits are still debated. The present study aimed to investigate the effect of HBO on brain damage in a rat model of transient focal cerebral ischemia and its underlying mechanism of action. Male Wistar rats, which had suffered 1.5h of transient middle cerebral artery occlusion (tMCAO) and had a Longa's neuron score of 3, were given pure oxygen at 3.0 atm absolute, for 60 min after the third hour of reperfusion. After 24h of reperfusion, rat brains were removed and studied. 2,3,5-triphenyltetrazolium chloride (TTC) and hematoxylin and eosin staining revealed that the infarct ratio in the HBO group increased remarkably when compared with the MCAO group. Up-regulation of extracellular signal-regulated kinase 1/2 (ERK1/2) activation was detected in the HBO group because of reactive oxygen species (ROS) generation. Autophagy appeared to be obstructed in the HBO group. Administration of the ERK1/2 inhibitor U0126 decreased the infarct ratio and improved protein clearance by autophagy in the HBO group. Collectively, these results suggest that HBO enlarges the area of brain damage via reactive oxygen species-induced activation of ERK1/2, which interrupts autophagy flux.

Mir-17-3p controls spinal neural progenitor patterning by regulating Olig2/Irx3 cross-repressive loop.

Neural patterning relies on transcriptional cross-repressive interactions that ensure unequivocal assignment of neural progenitor identity to proliferating cells. Progenitors of spinal motor neurons (pMN) and V2 interneurons (p2) are specified by a pair of cross-repressive transcription factors, Olig2 and Irx3. Lineage tracing revealed that many p2 progenitors transiently express the pMN marker Olig2 during spinal cord development. Here we demonstrate that the repression of Olig2 in p2 domain is controlled by mir-17-3p microRNA-mediated silencing of Olig2 mRNA. Mice lacking all microRNAs or just the mir-17∼92 cluster manifest a dorsal shift in pMN/p2 boundary and impairment in the production of V2 interneurons. Our findings suggest that microRNA-mediated repression of Olig2 mRNA plays a critical role during the patterning of ventral spinal progenitor domains by shifting the balance of cross-repressive interactions between Olig2 and Irx3 transcription factors.

The tumor suppressor DAPK is reciprocally regulated by tyrosine kinase Src and phosphatase LAR.

Death-associated protein kinase (DAPK) is a calmodulin-regulated serine/threonine kinase and elicits tumor suppression function through inhibiting cell adhesion/migration and promoting apoptosis. Despite these biological functions, the signaling mechanisms through which DAPK is regulated remain largely elusive. Here, we show that the leukocyte common antigen-related (LAR) tyrosine phosphatase dephosphorylates DAPK at pY491/492 to stimulate the catalytic, proapoptotic, and antiadhesion/antimigration activities of DAPK. Conversely, Src phosphorylates DAPK at Y491/492, which induces DAPK intra-/intermolecular interaction and inactivation. Upon EGF stimulation, a rapid Src activation leads to subsequent LAR downregulation, and these two events act in synergism to inactivate DAPK, thereby facilitating tumor cell migration and invasion toward EGF. Finally, DAPK Y491/492 hyperphosphorylation is found in human cancers in which Src activity is aberrantly elevated. These results identify LAR and Src as a DAPK regulator through their reciprocal modification of DAPK Y491/492 residues and establish a functional link of this DAPK-regulatory circuit to tumor progression.

Retinal ischemic injury rescued by sodium 4-phenylbutyrate in a rat model.

Retinal ischemia is a common cause of visual impairment for humans and animals. Herein, the neuroprotective effects of phenylbutyrate (PBA) upon retinal ischemic injury were investigated using a rat model. Retinal ganglion cells (RGCs) were retrograde labeled with the fluorescent tracer fluorogold (FG) applied to the superior collicoli of test Sprague-Dawley rats. High intraocular pressure and retinal ischemia were induced seven days subsequent to such FG labeling. A dose of either 100 or 400 mg/kg PBA was administered intraperitoneally to test rats at two time points, namely 30 min prior to the induction of retinal ischemia and 1 h subsequent to the cessation of the procedure inducing retinal ischemia. The test-rat retinas were collected seven days subsequent to the induction of retinal ischemia, and densities of surviving RGCs were estimated by counting FG-labeled RGCs within the retina. Histological analysis revealed that ischemic injury caused the loss of retinal RGCs and a net decrease in retinal thickness. For PBA-treated groups, almost 100% of the RGCs were preserved by a pre-ischemia treatment with PBA (at a dose of either 100 or 400 mg/kg), while post-ischemia treatment of RGCs with PBA did not lead to the preservation of RGCs from ischemic injury by PBA as determined by the counting of whole-mount retinas. Pre-ischemia treatment of RGCs with PBA (at a dose of either 100 or 400 mg/kg) significantly reduced the level of ischemia-associated loss of thickness of the total retina, especially the inner retina, and the inner plexiform layer of retina. Besides, PBA treatment significantly reduced the ischemia-induced loss of cells in the ganglion-cell layer of the retina. Taken together, these results suggest that PBA demonstrates a marked neuroprotective effect upon high intraocular pressure-induced retinal ischemia when the PBA is administered prior to ischemia induction.