Proteomic identification of differentially expressed and phosphorylated proteins in epidermis involved in larval-pupal metamorphosis of Helicoverpa armigera

<p>Abstract</p> <p>Background</p> <p>Metamorphosis is an important process in the life cycle of holometabolous insects and is regulated by insect hormones. During metamorphosis, the epidermis goes through a significant transformation at the biochemical and molecular levels.</p> <p>Results</p> <p>To identify proteins and phosphoproteins involved in this process, we separated and compared epidermal protein profiles between feeding larvae and metamorphically committed larvae using two-dimensional gel electrophoresis and Pro-Q Diamond Phosphoprotein Staining. Sixty-one spots showing differential expression and/or phosphorylation were analyzed by mass spectrometry and eighteen proteins were proved related to larval-pupal transformation. Eight of them were further examined at the mRNA level by Reverse Transcription Polymerase Chain Reaction (RT-PCR) and two of them were examined at the protein level by Western blot. Calponin was highly expressed in the metamorphic epidermis and phosphorylated by protein kinase C.</p> <p>Conclusion</p> <p>Our results suggest that the expression and phosphorylation of these proteins may play important roles in coordinating the biochemical processes involved in larval-pupal metamorphosis.</p

Affinity purification of recombinant human plasminogen activator from transgenic rabbit milk using a novel polyolresponsive monoclonal antibody

Purpose: To develop processes for effective isolation and purification of recombinant human plasminogen activator (rhPA) from transgenic rabbit milk.Methods: Immunoaffinity chromatography was selected and improved by a special polyol-responsive monoclonal antibody (PR-mAb). Alteplase was used as immunogen because of its similarity to rhPA in terms of structure. The PR-mAb was prepared by hybridoma technology and screened by ELISA-elution assay. Screening antibody was performed using rhPA milk in an ELISA-elution assay. The antibody clone C4-PR-mAb was selected for immunoaffinity chromatography. The rhPA was effectively bound to immobilized C4-PR-mAb on the column and was eluted with Tris buffer comprising 0.75 mol/L ammonium sulfate and 40n% propanediol (pH7.9). The rhPA was further purified by passing through Chromdex75 gel filtration column.Results: There were 12 hybridoma strains selected into the polyol responsive mAbs screen step and three hybridoma strains were superior for producing PR-mAbs (C1, C4, C8). The rhPA can be purified from transgenic rabbit milk and maintained a higher thrombolytic activity in vitro by FAPA.Conclusion: The results demonstrate the suitability of the alternative approach used in this study. Using immunoaffinity chromatography and  gel filtration column is feasible and convenient for extracting rhPA from milk, and should be useful for purifying other tPA mutants or other novel recombinant milkderived proteins.Keywords: tPA, Immunoaffinity chromatography, PR-mAb, ELISA-elution, Antibody, Thrombolytic activit

ELECTROACUPUNCTURING AT ZUSANLI POINT (ST36) ATTENUATES PRO-INFLAMMATORY CYTOKINE RELEASE AND ORGAN DYSFUNCTION BY ACTIVATING CHOLINERGIC ANTI-INFLAMMATORY PATHWAY IN RAT WITH ENDOTOXIN CHALLENGE

To investigate the effect of electro-acupuncturing (EA), at Zusanli point (ST36) on plasma cytokine release and organ dysfunction and their mechanism in conscious rats with endotoxin challenge. EA at Zusanli points obviously lowered the elevated levels of plasma TNF-α, and attenuated changes in parameters relevant to various organ functions at 2 h after LPS challenge. ɑ-BGT injection or bilateral cervical vagotomy could weaken or eliminate the effects of EA, and further aggravated the elevated levels of pro-inflammatory cytokines and organ dysfunction. The results suggested that EA at Zusanli points significantly reduced the release of pro-inflammatory cytokines and organ dysfunction after LPS challenge by activating cholinergic anti-inflammatory pathway

The Investigation of NOx Formation and Reduction during O2/CO2 Combustion of Raw Coal and Coal Char

AbstractThis study investigated the behavior of NOx emissions during combustion of pulverized coal char and pulverized raw coal in O2/CO2 environments under nearly isothermal condition in a drop tube furnace. A representative Chinese coal Ling Nancang (LNC) and LNC CO2-char (made in CO2 condition) were studied with four different excess oxygen ratios (α=0.6, 0.8, 1.2, and 1.4). Combustion experiments were performed with an elevated concentration (850ppm) of NO. When α increased from 0.6 to 1.4, the total amount of NOx reduction was increased in both pulverized coal and char combustion. To investigate the effects of coal and char on NOx reburn reactions, LNC coal char and the raw coal combustion experiments were also performed under three high background NO concentrations (400, 850, and 1200ppm) in O2/CO2 atmosphere. During experiments with elevated NO concentrations, there was an increase in net NO reduction; however, there was also a decrease in the calculated NO reduction ratio

Characteristic Analysis from Excessive to Deficient Syndromes in Hepatocarcinoma Underlying miRNA Array Data

Traditional Chinese medicine (TCM) treatment is regarded as a safe and effective method for many diseases. In this study, the characteristics among excessive, excessive-deficient, and deficient syndromes of Hepatocellular carcinoma (HCC) were studied using miRNA array data. We first calculated the differentially expressed miRNAs based on random module t-test and classified three TCM syndromes of HCC using SVM method. Then, the weighted miRNA-target networks were constructed for different TCM syndromes using predicted miRNA targets. Subsequently, the prioritized target genes of upexpression network of TCM syndromes were analyzed using DAVID online analysis. The results showed that there are distinctly different hierarchical cluster and network structure of TCM syndromes in HCC, but the excessive-deficient combination syndrome is extrinsically close to deficient syndrome. GO and pathway analysis revealed that the molecular mechanisms of excessive-deficient and deficient syndromes of HCC are more complex than excessive syndrome. Furthermore, although excessive-deficient and deficient syndromes have similar complex mechanisms, excessive-deficient syndrome is more involved than deficient syndrome in development of cancer process. This study suggested that miRNAs might be important mediators involved in the changing process from excessive to deficient syndromes and could be potential molecular markers for the diagnosis of TCM syndromes in HCC