The maintenance of pure primary lymphocytes culture for long periods may be difficult because of its inability to divide continuously. In addition, lymphocytes separation methods such as Ficoll-Paque, RBC lysis and immunomagnetic microbeads separation may have some affect on cell viability. The objective of this study is to determine various types of lymphocytes purification methods, in order to prolong primary lymphocytes culture to 72 hours. The second objective is to use these primary lymphocytes as targets for quantitative and qualitative cell viability assays when analysing the action of toxins isolated from natural products. Human blood was drawn and purified by using Ficoll-Paque, RBC lysis or immunomagnetic separation column method in various combinations. The purified lymphocytes were also grown with and without the growth enhancement factor, concanavalin-A. Cell viability assays were carried out for 72 hours at 24 hours interval. The lymphocytes purified using RBC lysis method, with or without concanavalin-A can prolong 100% cell viability for 72 hours whilst lymphocytes purified using Ficoll-Paque and supplemented with concanavalin-A showed an increase in cell viability of over 250% at 72 hours incubation. It was observed only lymphocytes purified using Ficoll-Paque followed by the immunomagnetic microbeads separation method and supplemented with concanavalin-A showed overall cell viability increase, reaching 300% at 72 hours incubation. This method was a reliable model to test the cytotoxicity of the Bacillus thuringiensis parasporal inclusion, suggesting that the method achieves the objectives of the study.

Child ; Humans ; Lung Diseases, Interstitial/diagnostic imaging*

Methods: Forty AIS patients were recruited. Demographic, preoperative, and postoperative data were recorded. The magnitude and characteristics of postoperative pain were assessed using the painDETECT questionnaire through telephone enquiries at intervals of 2, 6, 12, and 24 weeks. Statistical analyses were followed by Pearson correlation test to determine the relationship between pain scores at 6, 12, and 24 weeks with the risk factors. Results: Based on the painDETECT questionnaire, 90% of the patients had nociceptive pain, and 10% had a possible neuropathic pain component at 2 weeks postoperatively as per a mean painDETECT score of 7.1±4.5. Assessments at 6, 12, and 24 weeks showed that no patients had neuropathic pain with painDETECT scores of 4.4±3.2, 2.9±2.9, and 1.5±2.0, respectively. There was a significant correlation between total postoperative morphine use during 48 hours after the surgery and a tendency to develop neuropathic pain (p=0.022). Conclusions Chronic neuropathic pain was uncommon in AIS patients who had undergone PSF surgery. Higher opioid consumption will increase the possibility of developing chronic neuropathic pain.

Introduction: This study looked into the different anatomical locations of pain and their trajectories within the first two weeks after Posterior Spinal Fusion (PSF) surgery for Adolescent Idiopathic Scoliosis (AIS). Methods: We prospectively recruited patients with Adolescent Idiopathic Scoliosis (AIS) scheduled for PSF surgery. The anatomical locations of pain were divided into four: (1) surgical wound pain; (2) shoulder pain; (3) neck pain; and (4) low back pain. The anatomical locations of pain were charted using the visual analogue pain score at intervals of 12, 24, 36, 48 hours; and from day-3 to -14. Patient-controlled analgesia (morphine), use of celecoxib capsules, acetaminophen tablets and oxycodone hydrochloride capsule consumption were recorded. Results: A total of 40 patients were recruited. Patients complained of surgical wound pain score of 6.2±2.1 after surgery. This subsequently reduced to 4.2±2.0 by day-4, and to 2.4±1.3 by day-7. Shoulder pain scores of symptomatic patients peaked to 4.2±2.7 at 24 hours and 36 hours which then reduced to 1.8±1.1 by day-8. Neck pain scores of symptomatic patients reduced from 4.2±1.9 at 12 hours to 1.8±1.1 by day-4. Low back pain scores of symptomatic patients reduced from 5.3±2.3 at 12 hours to 1.8±1.1 by day12. Conclusions: Despite the presence of different anatomical locations of pain after surgery, surgical wound was the most significant pain and other anatomical locations of pain were generally mild. Surgical wound pain reduced to a tolerable level by day-4 when patients can then be comfortably discharged. This finding provides useful information for clinicians, patients and their caregivers.

INTRODUCTIONTranscatheter aortic valve implantation (TAVI) is an effective treatment for high-risk or inoperative patients with severe aortic stenosis. Given the unique characteristics of Asian populations, questions regarding mid-term outcomes in Asians undergoing TAVI have yet to be addressed. We evaluated the two-year clinical outcomes of TAVI in an Asian population using Valve Academic Research Consortium-2 definitions.

METHODSThis prospective study recruited 59 patients from a major academic medical centre in Singapore. The main outcomes were two-year survival rates, peri-procedural complications, symptom improvement, valvular function and assessment of learning curve.

RESULTSMean age was 76.8 years (61.0% male), mean body surface area 1.6 mand mean logistic EuroSCORE 18.7%. Survival was 93.2%, 86.0% and 79.1% at 30 days, one year and two years, respectively. At 30 days post TAVI, the rate of stroke was 1.7%, life-threatening bleeding 5.1%, acute kidney injury 25.0%, major vascular complication 5.1%, and new permanent pacemaker implantation 6.8%. 29.3% of TAVI patients were rehospitalised (47.1% cardiovascular-related) within one year. These composite outcomes were measured: device success (93.2%); early safety (79.7%); clinical efficacy (66.1%); and time-related valve safety (84.7%). Univariate analysis found these predictors of two-year all-cause mortality: logistic EuroSCORE (hazard ratio [HR] 1.07; p < 0.001); baseline estimated glomerular filtration rate (HR 0.97; p = 0.048); and acute kidney injury (HR 5.33; p = 0.022). Multivariate analysis identified non-transfemoral TAVI as a predictor of cardiovascular-related two-year mortality (HR 14.64; p = 0.008).

CONCLUSIONDespite the unique clinical differences in Asian populations, this registry demonstrated favourable mid-term clinical and safety outcomes in Asians undergoing TAVI.