Cloning and Expression of Pseudomonas aeruginosa AlkB Gene in E. coli

Pre identified hydrocarbons degrading bacteria were used in this study, specific primer was conducted to amplification of AlkB gene, approximately 1206bp band size of this gene for Pseudomonas aeruginosa was detected and proofed by sequence and alignment analysis with NCBI database. The AlkB gene was inserted in PET-21a(+) plasmid vector as expression vector, then transformed in BL21(DE3) competent E. coli and confirmed by colony PCR technique using the T7 promoter and T7 terminator primers. The expression of the inserted gene was checked by determined the concentration of AlkB protein for multiple periods by Bradford assay method and the SDS-polyacrylamide gel electrophoresis method was revealed band of ~46 KD molecular weight of the concerned protein. The gene amplification and cloning strategy was lay out before the practical part of the study by SnapGene software, this study was conducted to introduce cloned bacteria which facilitate the first step (key step) of alkane’s biodegradation and propose an appropriate strategy to construct genetically engineered microorganisms with multiple recombinant plasmid for enhance the degradation of the aliphatic fraction of hydrocarbo

Comparison between CURB-65, PSI, and SIPF scores as predictors of ICU admission and mortality in community-acquired pneumonia

Objective: To compare the prognostic value of the SIPF (shock index and hypoxemia) score as the combination of shock index (heart rate/systolic blood pressure) >0.7 (1 point) plus PaO2/FiO2 < 250 (1 point), and the severity score for community-acquired pneumonia (CAP) (CURB-65) and pneumonia severity index (PSI) in predicting the need for ICU admission and mortality of patients with community-acquired pneumonia. Patients and methods: This retrospective study was conducted on patients with CAP admitted to AL-Hussein University hospital (Egypt), Muhayl general hospital and King Khalid hospital at Hail, (KSA). The information required for calculating SIPF, PSI and CURB-65 was extracted from the medical records. Results: We studied 100 patients with community-acquired pneumonia (64 men, 36 women). Thirty-four patients needed ICU admission (while 66 did not need ICU admission and admitted in observation room or general ward) and among the ICU patients 21 cases needed mechanical ventilation. Ten cases died; 9 cases in ICU and one case in observation room (ward). The ability to predict ICU admission was higher for SIPF score compared to CURB-65 (AUC SIPF 0.88 vs. 0.83; p < 0.001) and PSI (AUC SIPF 0.88 vs. 0.79; p < 0.001). The ability to predict mortality was higher for SIPF score compared to CURB-65(AUC SIPF 0.80 vs. 0.84; p < 0.001) and PSI (AUC SIPF 0.80 vs. 0.83; p < 0.001). Conclusion: The ability of SIPF score to predict ICU admission in CAP is higher than that of CURB-65 and PSI. Simple SIPF score could be a useful tool to predict mortality in CAP