Cloning and expression of a codon-optimized gene encoding the infl uenza A virus nucleocapsid protein in Lactobacillus casei

Lactic acid bacteria (LAB) species are envisioned as promising vehicles for the mucosal delivery of therapeutic and prophylactic molecules, including the development of oral vaccines. In this study, we report on the expression of a synthetic nucleocapsid (NP) gene of infl uenza A virus in Lactobacillus casei. The NP gene was re-designed based on the tRNA pool and the codon usage preference of L. casei BL23. The codon-optimized NP gene was then cloned and expressed in L. casei RCEID02 under the control of a constitutive promoter, that of the lactate dehydrogenase (ldh) gene. The synthetic NP gene was further expressed in L. casei EM116 under the control of an inducible promoter, that of the structural gene of nisin (nisA) from Lactococcus lactis. Based on Western blot analysis, the specifi c protein band of NP, with a molecular mass of 56.0 kDa, was clearly detected in both expression systems. Thus, our study demonstrates the success of expressing a codon-optimized infl uenza A viral gene in L. casei. The suitability of the recombinant LAB strains for immunization purposes is currently under evaluation. [Int Microbiol 2013; 16(2):93-101]Keywords: Lactobacillus casei; lactic acid bacteria; infl uenza A virus; viral nucleocapsid proteins; heterologous expression; codon usag

Exploration of pain management following implementation of WHO pain guidelines

19646 Background: Pain is among the most common symptoms encountered in cancer patients and remains the first priority of care. Methods: This cross sectional study aimed to explore a pattern of pain management at KKU Hospital by utilizing a numeric rating scale (0–10). Cancer pain patients were categorized based on prior analgesic exposure into two groups; Naïve group, and Routine group. Treatments were defined according to WHO as 1) drug treatment relevant to pain severity, 2) analgesics being prescribed as around-the- clock and 3) analgesics used for break-through pain for patients receiving strong opioid. Results: From Dec 2005 to Jul 2006, 261 patients were enrolled. 93.1% (n=243) were in advanced stages and 88.5% (n=231) were in moderate to severe pain. This pain interfered with patient’s daily life activities mildly to moderately as each pain score increased (p-value&lt;0.01). In Naive group (n=159), 32.7% (n=52) were given analgesics following the WHO on both days 1 and day 3 of admission whereas 40.2% (n=64) patients were not. A decreased pain score was greater (2.61, SD±1.5) in a group following the WHO on day 1. Additionally, a decreased pain score was greater (3.91, SD±1.8) in a group following the WHO on day 3 (p-value &lt;0.0001). This pain score decreased was also clinically significant as pain score reduced more than 3 points. In Routine group (n=102), 32 (31.4%) were given analgesics following the WHO guideline on both day 1 and day 3 of admission. In contrast, 36 (35.3%) were not. A decreased pain score was greater (2.59, SD±1.8) in a routine group following the WHO on day 1. Moreover, a decreased pain score was greater (3.95, SD±1.8) in a group following the WHO on day 3. The clinical significance of pain score reduced was also found on day 3. Of the 261 evaluable patients, the pattern of analgesics usage following the WHO guideline was increased in both groups comparing to at the beginning of the study. Conclusions: The results demonstrated that patients who received pain management following the WHO guideline reported significantly lower pain intensity than those not following the WHO. No significant financial relationships to disclose. </jats:p