Purification, biochemical characterization, and molecular cloning of cellulase from Bacillus licheniformis strain Z9 isolated from soil
Background: Cellulose is the most prevalent biomass and renewable energy source in nature. The hydrolysis of cellulosic biomass to glucose units is essential for the economic exploitation of this natural resource. Cellulase enzyme, which is largely generated by bacteria and fungus, is commonly used to degrade cellulose. Cellulases are used in a variety of industries, including bioethanol manufacturing, textiles, detergents, drugs, food, and paper. As part of our quest to find an efficient biocatalyst for the hydrolysis of cellulosic biomass, we describe the amplification, cloning, and sequencing of cellulase (cel9z) from Bacillus licheniformis strain Z9, as well as the characterization of the resulting enzyme.
Results: Cellulase was partially purified from B. licheniformis strain Z9 using (NH4)2SO4 precipitation and Sephadex G-100 gel column chromatography with 356.5 U/mg specific activity, 2.1-purification fold, and 3.07 % yield. The nucleotide sequence of the cellulase gene was deposited to the GenBank, B. licheniformis strain Z9 cellulase (cel9z) gene, under accession number MK814929. This corresponds to 1453 nucleotides gene and encodes for a protein composed of 484 amino acids. Comparison of deduced amino acids sequence to other related cellulases showed that the enzyme cel9z can be classified as a glycoside hydrolase family 9. SDS-PAGE analysis of the purified enzyme revealed that the molecular mass was 54.5 kDa. The optimal enzyme activity was observed at pH 7.4 and 30 °C. The enzyme was found to be strongly inhibited by Mg2+ and Na+, whereas strongly activated by Fe3+, Cu2+, and Ca2+.
Conclusions: B. licheniformis strain Z9 and its cellulase gene can be further utilized for recombinant production of cellulases for industrial application.
Biochemical and subcellular characterization of a squid hnRNPA/B-like protein 2 in osmotic stress activated cells reflects molecular properties conserved in this protein family
Background: We have identified endogenous p65 to be an SDS-stable dimer protein composed of ~ 37 kDa hnRNPA/B-like subunits. We have investigated molecular properties involved in the stability of dimeric form, and their regulation in the transition between monomeric and dimeric forms of hnRNPA/B-like protein 2. We also investigated a cellular property conserved between squid hnRNPA/B-like protein 2 and human hnRNPA1 protein in a neuronal context.
Methods and results: Here we show biochemical properties of a recombinant hnRNPA/B-like protein 2 (rP2) in vitro experiments, as one of p65 subunit. We found that interaction between rP2 and RNA molecules interfered with the dynamics of rP2 dimers formation, involved in disulfide bonds and/or postranslational alterations in distinct stage of SDS-stable dimers formation. In addition, we have performed immunofluorescence in SH-SY5Y cells and observed that the pEGFP-P2 fusion protein was expressed in the nucleus, similar to what is observed for human hnRNPA1 protein.
Conclusion: Our results reinforce the idea that p65 is an SDS-stable dimer. Thus, a deeper understanding between monomeric and dimeric transition dynamic is critical into evolution of several neurodegenerative disease.
A quality of life, clinical and biochemical improvements after catheter ablation of persistent arrhythmia in patients with structural heart disease and arrhythmia-mediated cardiomyopathy
Background: Arrhythmia-mediated cardiomyopathy (AMC) is an essential clinical situation which is commonly underdiagnosed. Successful arrhythmia control leads to improvement in health-related quality of life (HRQoL) and heart failure (HF) symptoms in patients with structural heart disease (SHD).
Aims: The aim of the study was to evaluate the impact of catheter ablation (CA) of persistent arrhythmia on HRQoL, biochemical and clinical parameters HF in patients with SHD and AMC.
Methods: Patients with SHD, on optimal medical treatment, with persistent arrhythmia and strong suspicion of AMC, scheduled for CA were prospectively enrolled. Study procedures included: HRQoL measurement (Minnesota Living With Heart Failure Questionnaire [MLHFQ] and EuroQol Research Foundation [EQ-5D-3L] questionnaire), biomarkers (N-terminal pro-B-type natriuretic peptide [NT-proBNP], troponin T [TnT], matrix metaloproteinase-9 [MMP-9], soluble suppression of tumorigenesis-2 [sST2], tissue inhibitor of matrix metalloproteinase-1 [TIMP-1]), transthoracic echocardiography and clinical assessment.
Results: At 6 months, 30/35 (86%) patients were free of persistent arrhythmia. Patients who underwent successful CA had a significant improvement in HRQoL: MLHFQ (median [interquartile range, IQR], -22 [-28;-11]; P <0.001), EQ5D-3L Score (mean [standard deviation], 21.8 (16.8); P <0.001); EQ5D-3L index (median [IQR], 0.09 (0.05;0.18); p <0.001). A significant decrease in injury biomarkers: NT-proBNP (median [IQR], -414 [-1397;-318] pg/ml; P <0.001), TnT (median [IQR], -2.27 (-8.52;0.55) ng/ml; P <0.01) but not fibrosis biomarkers (median [IQR], sST2: 2.20 [-5.4;4.3] ng/ml; P = 0.741, MMP-9: 34 [-376;283] ng/ml; P = 0.881, TIMP-1: 11.1 [-17.1;31.9] ng/ml; P = 0.215) was observed. There was a significant increase of left ventricular ejection fraction (LVEF) (mean [SD], 9.8 [5.9]; P <0.01).
Conclusions: Successful CA significantly improved clinical status, LVEF and HRQoL of patients with SHD and AMC.
Clinical and Biological Significance of DNA Methylation-Driven Differentially Expressed Genes in Biochemical Recurrence After Radical Prostatectomy
Background: Biochemical recurrence (BCR) after radical prostatectomy indicates poor prognosis in patients with prostate cancer (PCA). DNA methylation (DNAm) is a critical factor in tumorigenesis and has attracted attention as a biomarker for the diagnosis, treatment, and prognosis of PCA. However, the predictive value of DNAm-derived differentially expressed genes (DMGs) in PCA with BCR remains elusive.
Methods: We filtered the methylated genes and the differentially expressed genes (DGEs) for more than 1,000 clinical samples from the TCGA cohort using the chAMP and DESeq2 packages of R language, respectively. Next, we integrated the DNAm beta value and gene expression data with the Mithymix package of R language to obtain the DMGs. Then, 1,000 times Cox LASSO regression with 10-fold cross validation was performed to screen signature DMGs and establish a predictive classifier. Univariate and multivariate cox regressive analyses were used to identify the prognostic factors to build a predictive model, and its performance was measured by receiver operating characteristic, calibration curves, and Harrell’s concordance index (C-index). Additionally, a GEO dataset was used to validate the prognostic classifier.
Results: One hundred DMGs were mined using the chAMP and Methymix packages of R language. Of these, seven DMGs (CCK, CD38, CYP27A1, EID3, HABP2, LRRC4, and LY6G6D) were identified to build the prognostic classifier (Classifier) through LASSO analysis. Moreover, univariate and multivariate Cox regression analysis determined that the Classifier and pathological T stage (pathological_T) were independent predictors of BCR (hazard ratio (HR 2.2), (95% CI 1.4-3.5), p < 0.0012, and (HR 1.8), (95% CI 1.0-3.2), p < 0.046). A nomogram based on the Classifier was constructed, with high prediction accuracy for BCR-free survival in TCGA and GEO datasets. GSEA enrichment analysis showed that the DMGs were mainly enriched in the metabolism pathways.
Conclusion: We identified and validated the nomogram of BCR-free survival for PCA patients, which has the potential to guide treatment decisions for patients at differing risks of BCR. Our study deepens the understanding of DMGs in the pathogenesis of PCA.
Biochanin A |
|||
| 540046 | MedKoo Biosciences | 200.0mg | 225 EUR |
Biochanin A |
|||
| B387250 | Toronto Research Chemicals | 100mg | 63 EUR |
Biochanin A |
|||
| B10821 | Pfaltz & Bauer | 500MG | 779.11 EUR |
Biochanin A |
|||
| GP7014 | Glentham Life Sciences | 1g | 108.65 EUR |
Biochanin A |
|||
| GP7014-1 | Glentham Life Sciences | 1 | 118.7 EUR |
Biochanin A |
|||
| GP7014-1G | Glentham Life Sciences | 1 g | 180 EUR |
Biochanin A |
|||
| HY-14595 | MedChemExpress | 10mM/1mL | 107.14 EUR |
Biochanin A |
|||
| MBS3604647-100mg | MyBiosource | 100mg | 200 EUR |
Biochanin A |
|||
| MBS3604647-200mg | MyBiosource | 200mg | 210 EUR |
Biochanin A |
|||
| MBS3604647-5x200mg | MyBiosource | 5x200mg | 630 EUR |
Biochanin A |
|||
| MBS576412-100mg | MyBiosource | 100mg | 120 EUR |
Biochanin A |
|||
| MBS576412-200mg | MyBiosource | 200mg | 145 EUR |
Biochanin A |
|||
| MBS576412-500mg | MyBiosource | 500mg | 160 EUR |
Biochanin A |
|||
| MBS576412-5x500mg | MyBiosource | 5x500mg | 575 EUR |
Biochanin A |
|||
| N1308-200 | ApexBio | 200 mg | 40 EUR |
Biochanin A |
|||
| N1308-5.1 | ApexBio | 10 mM (in 1mL DMSO) | 40 EUR |
Biochanin A |
|||
| TB0167-0200 | ChemNorm | 5X20mg | 328.8 EUR |
Biochanin B |
|||
| TB0216-1000 | ChemNorm | 8XX20mg | 328.8 EUR |
Biochanin A |
|||
| T2859-10mg | TargetMol Chemicals | 10mg | Ask for price |
Biochanin A |
|||
| T2859-1g | TargetMol Chemicals | 1g | Ask for price |
Biochanin A |
|||
| T2859-1mg | TargetMol Chemicals | 1mg | Ask for price |
Biochanin A |
|||
| T2859-50mg | TargetMol Chemicals | 50mg | Ask for price |
Biochanin A |
|||
| T2859-5mg | TargetMol Chemicals | 5mg | Ask for price |
Biochrom EZ Read 400 ELISA |
|||
| MIC8400 | Scientific Laboratory Supplies | EACH | 4775.46 EUR |
Biochemical Test Kit - I |
|||
| 92875 | Sisco Laboratories | 1 Kit | 20.53 EUR |
Biochemical Test Kit - II |
|||
| 65896 | Sisco Laboratories | 1 Kit | 18.82 EUR |
Biochemical changes in patients during hypothyroid phase after thyroidectomy
- Hypothyroidism is the most common long-term consequence after total thyroidectomy. The objective of the present study was to evaluate the lipid profile and liver function in patients after hypothyroidism. Sixty patients who underwent a surgical operation to remove thyroid were included in this study, and thirty healthy subjects were used as a control. The study was conducted at Al-Sadr Medical City in Al-Najaf city, in Iraq, from October 2020 to March 2021. Thyroid-stimulating hormone (TSH) was very high in patients at a hypothyroid stage after hypothyroidism. The results showed a significant increase in total cholesterol, triglycerides (TG), low-density lipoprotein (LDL), and the ratio of total cholesterol/high-density lipoprotein (HDL).
- The study also revealed a significant increase in the liver enzymes aspartate aminotransferase (AST) and alanine transaminase (ALT) and a significant decrease in alkaline phosphatase (ALP) in patients with thyroidectomy compared to the control group. The correlation matrix revealed a strong positive correlation between TSH levels and total cholesterol, triglycerides, LDL, AST, ALT, and ALP. It was concluded that hypothyroidism, the major consequence of thyroidectomy, causes dysfunction in lipid metabolism and liver enzymes resulting in secondary hyperlipidemia and liver dysfunction.