Effects of Probiotic Bacteria Bacillus on Growth Performance,Digestive Enzyme Activity,and Hematological Parameters of Asian Sea Bass,Lates calcarifer (Bloch)

Probiotics and Antimicrobial Proteins - This study was conducted to evaluate different doses of two species of Bacillus (Bacillus licheniformis and Bacillus subtilis), on growth parameters,...     

Enzymatic pre‐treatment of microalgae cells for enhanced extraction of proteins

Crude proteins and pigments were extracted from different microalgae strains, both marine and freshwater. The effectiveness of enzymatic pre‐treatment prior to protein extraction was evaluated and compared to conventional techniques, including ultrasonication and high‐pressure water extraction. Enzymatic pre‐treatment was chosen as it could be carried out at mild shear conditions and does not subject the proteins to high temperatures, as with the ultrasonication approach. Using enzymatic pre‐treatment, the extracted proteins yields of all tested microalgae strains were approximately 0.7 mg per mg of dry cell weight. These values were comparable to those achieved using a commercial lytic kit. Ultrasonication was not very effective for proteins extraction from Chlorella sp., and the extracted proteins yields did not exceed 0.4 mg per mg of dry cell weight. For other strains, similar yields were achieved by both treatment methods. The time‐course effect of enzymatic incubation on the proteins extraction efficiency was more evident using laccase compared to lysozyme, which suggested that the former enzyme has a slower rate of cell disruption. The crude extracted proteins were fractionated using an ion exchange resin and were analyzed by the electrophoresis technique. They were further tested for their antioxidant activity, the highest of which was about 60% from Nannochloropsis sp. The total phenolic contents in the selected strains were also determined, with Chlorella sp. showing the highest content reaching 17 mg/g. Lysozyme was also found to enhance the extraction of pigments, with Chlorella sp. showing the highest pigments contents of 16.02, 4.59 and 5.22 mg/g of chlorophyll a, chlorophyll b and total carotenoids, respectively.     

Evaluation of Macroscopic and Microscopic Components of Partial Resistance to Leaf Rust in Durum Wheat

Leaf rust, caused by the fungus Puccinia triticina, is considered one of the most important foliar diseases in durum wheat. Hypersensitive resistance (HR) may be rapidly overcome by the pathogen when resistant cultivars are grown on a large acreage or following changes in virulence in the pathogen population. Prolonging the durability of the resistance requires uses of other types of resistance such as partial resistance (PR). In this study, six durum wheat lines provided by the International Center for Corn and Wheat Improvement (CIMMYT) with a high level of PR to leaf rust were studied in monocyclic tests in a growth chamber. Inoculations were performed on both primary and fifth leaves using the Spanish race DGB/BN. UV fluorescence microscopy was employed to determine microscopic components of the resistance, such as the number of early aborted infection units not associated with plant cell necrosis (EA?) and relative colony size (RCS) of the established infection units. Macroscopic components of PR such as latency period, infection frequency and uredinium size were measured as well. All six resistant lines were characterized by a higher EA? and smaller RCS respect to the susceptible control ‘Don Rafael’. Line 3 showed the highest level of PR. It had 22% of EA? compared with 4% in the susceptible control, and the smallest RCS (17% respect to RCS of ‘Don Rafael’) at adult plant stage. Both EA? and RCS had a high heritability (more than 97%) and the correlation with macroscopic parameters (latency period and uredinium size) was also high (significant at 0.001 level). Hence, PR to leaf rust in these durum wheat genotypes has been revealed at microscopic level (higher EA? and smaller RCS).     

Optimisation of a cost-effective culture medium for the large-scale production of Bacillus subtilis UTB96

Fermentation optimisation to achieve high biomass and high efficiency of a biocontrol product is millstone in biocontrol sciences. Here, a Placket–Burman design used for comparison and screening of some environmental factors that are effective on bacterial biomass of Bacillus subtilis UTB96. A response surface methodology used to determine the optimal points for three factors including pH, temperature and C/N ratio where the biomass is high. The results of response surface methodology application showed that the optimum conditions for maximum production of biomass in the medium occurs at pH 7, temperature 30?°C and C/N ratio of 23:1. Bacteria derived from optimised conditions either from laboratory or semi-industrial bioreactors, showed a considerable increase in biomass and also, their antagonistic activity against Phytophthora drechsleri in a plate assay. However, optimisation of culture medium in a laboratory bioreactor decreased the antagonistic activity against Aspergillus flavus. Application of the optimised culture medium in both semi-industrial and laboratory bioreactors reduced the length of the lag phase of bacterial growth.     

Long-Term Outcome of Anti-Glomerular Basement Membrane Antibody Disease Treated with Immunoadsorption

Background

Anti-glomerular basement membrane (GBM) antibody disease may lead to acute crescentic glomerulonephritis with poor renal prognosis. Current therapy favours plasma exchange (PE) for removal of pathogenic antibodies. Immunoadsorption (IAS) is superior to PE regarding efficiency of antibody-removal and safety. Apart from anecdotal data, there is no systemic analysis of the long-term effects of IAS on anti-GBM-disease and antibody kinetics.

Objective

To examine the long-term effect of high-frequency IAS combined with standard immunosuppression on patient and renal survival in patients with anti-GBM-disease and to quantify antibody removal and kinetics through IAS.

Design

Retrospective review of patients treated with IAS for anti-GBM-antibody disease confirmed by biopsy and/or anti-GBM-antibodies.

Setting

University Hospital of Vienna, Austria.

Participants

10 patients with anti-GBM-disease treated with IAS.

Measurements

Patient and renal survival, renal histology, anti-GBM-antibodies.

Results

Anti-GBM-antibodies were reduced by the first 9 IAS treatments (mean number of 23) to negative levels in all patients. Renal survival was 40% at diagnosis, 70% after the end of IAS, 63% after one year and 50% at the end of observation (mean 84 months, range 9 to 186). Dialysis dependency was successfully reversed in three of six patients. Patient survival was 90% at the end of observation.

Conclusion

IAS efficiently eliminates anti-GBM-antibodies suggesting non-inferiority to PE with regard to renal and patient survival. Hence IAS should be considered as a valuable treatment option for anti-GBM-disease, especially in patients presenting with a high percentage of crescents and dialysis dependency due to an unusual high proportion of responders.     

Synergistic Anti-Cancer Activity of the Combination of 1,25-Dihydroxyvitamin D3 and Retinoic Acid in U937 Cell Line

Background:MicroRNA is a form of non-coding RNAs that able to regulate gene expression. miR-424 is one of the members of the regulatory family, which plays an important role in the proliferation and differentiation of myeloid cells. Epigenetic changes can change the level of miR-424 under environmental factors. Therefore, the level of expression of miR-424 in U937 cells of the myeloid line was evaluated in this research under the influence of vitamin D3 (VitD3) and retinoic acid (RA).Methods:In this study, U937 cells were cultured in the presence of VitD3, and RA to evaluate cell proliferation, viability via the trypan blue exclusion test, and expression level of miR-424 by real-time PCR at specific times.Results:Cell proliferation has shown a significant decrease in the RA group versus other groups during incubation times (P < 0.05). In VitD3 group, there was a significant increase in cell proliferation after 24- and 48-hours incubation periods versus other groups. In the VitD3 and RA groups, the increase of cell proliferation caused the downregulation of miR-424. In addition, the upregulation of VitD3 group and downregulation of the RA group were significant versus the control group (P < 0.05).DiscussionWe concluded that the expression level of miR-424 was critically affected in the dose- and time-dependent of RA and VitD3 treatment in the U937 cell line. Treatment with VitD3 decreased the expression of miR-424 and RA treatment increase miR-424 expression level in physiological doses.Key Words: Cell Proliferation, Differentiation, miR-424, U937 Cells     

Fabrication and evaluation of aptamer-conjugated paclitaxel-loaded magnetic nanoparticles for targeted therapy on breast cancer cells

Molecular Biology Reports - Targeted drug delivery vehicles make it possible to deliver anti-cancer drugs to the cells or tissues of interest. Aptamers are peptide or oligonucleotide molecules that...     

Sternal Wound Infection after Cardiac Surgery: Management and Outcome

Background

Sternal Wound Infection (SWI) is a severe complication after cardiac surgery. Debridement associated with primary closure using Redon drains (RD) is an effective treatment, but data on RD management and antibiotic treatment are scarce.

Methods

We performed a single-center analysis of consecutive patients who were re-operated for SWI between 01/2009 and 12/2012. All patients underwent a closed drainage with RD (CDRD). Patients with endocarditis or those who died within the first 45 days were excluded from management analysis. RD fluid was cultured twice weekly. Variables recorded were clinical and biological data at SWI diagnosis, severity of SWI based on criteria for mediastinitis as defined by the Centers for Disease Control (CDC), antibiotic therapy, RD management and patient’s outcome.

Results

160 patients developed SWI, 102 (64%) fulfilled CDC criteria (CDC+) and 58 (36%) did not (CDC- SWI). Initial antibiotic treatment and surgical management were similar in CDC+ and CDC- SWI. Patients with CDC+ SWI had a longer duration of antibiotic therapy and a mortality rate of 17% as compared to 3% in patients with CDC- SWI (p = 0.025). Rates of superinfection (10% and 9%) and need for second reoperation (12% and 17%) were similar. Failure (death or need for another reoperation) was associated with female gender, higher EuroScore for prediction of operative mortality, and stay in the ICU.

Conclusion

In patients with SWI, initial one-stage surgical debridement with CDRD is associated with favorable outcomes. CDC+ and CDC- SWI received essentially the same management, but CDC+ SWI has a more severe outcome.     

Status of Serum Copper,Magnesium, and Total Antioxidant Capacity in Patients with Polycystic Ovary Syndrome

Biological Trace Element Research - This study evaluates serum copper and magnesium and total antioxidant capacity levels in PCOS patients. In this regard, the probable association of copper and...     

Mitotic Arrest and Apoptosis in Breast Cancer Cells Induced by Origanum majorana Extract: Upregulation of TNF-α and Downregulation of Survivin and Mutant p53

Background

In the present study, we investigated the effect of Origanum majorana ethanolic extract on the survival of the highly proliferative and invasive triple-negative p53 mutant breast cancer cell line MDA-MB-231.

Results

We found that O. majorana extract (OME) was able to inhibit the viability of the MDA-MB-231 cells in a time- and concentration-dependent manner. The effect of OME on cellular viability was further confirmed by the inhibition of colony growth. We showed, depending on the concentration used, that OME elicited different effects on the MDA-MB 231 cells. Concentrations of 150 and 300 µg/mL induced an accumulation of apoptotic–resistant population of cells arrested in mitotis and overexpressing the cyclin-dependent kinase inhibitor, p21 and the inhibitor of apoptosis, survivin. On the other hand, higher concentrations of OME (450 and 600 µg/mL) triggered a massive apoptosis through the extrinsic pathway, including the activation of tumor necrosis factor-α (TNF-α), caspase 8, caspase 3, and cleavage of PARP, downregulation of survivin as well as depletion of the mutant p53 in MDA-MB-231 cells. Furthermore, OME induced an upregulation of γ-H2AX, a marker of double strand DNA breaks and an overall histone H3 and H4 hyperacetylation.

Conclusion

Our findings provide strong evidence that O. majorana may be a promising chemopreventive and therapeutic candidate against cancer especially for highly invasive triple negative p53 mutant breast cancer; thus validating its complementary and alternative medicinal use.