METHODS: A total of 71 malaria microscopy positive blood samples collected in blood spots were obtained from the Sarawak State Health Department. Using 18s rRNA as the target gene, nested PCR and SYBR green I LAMP assay were performed following the DNA extraction. The colour changes of LAMP end products were observed by naked eyes.
RESULTS: LAMP assay demonstrated a detection limit of 10 copies/µL in comparison with 100 copies/µL nested PCR. Of 71 P. knowlesi blood samples collected, LAMP detected 69 microscopy-positive samples. LAMP exhibited higher sensitivity than nested PCR assay. The SYBR green I LAMP assay was 97.1% sensitive (95% CI 90.2-99.7%) and 100% specific (95% CI 83.2-100%). Without opening the cap, incorporation of SYBR green I into the inner cap of the tube enabled the direct visualization of results upon completion of amplification. The positives instantaneously turned green while the negatives remained orange.
CONCLUSIONS: These results indicate that SYBR green I LAMP assay is a convenient diagnosis tool for the detection of P. knowlesi in remote settings.
METHODS: In a double blind, randomised crossover design, 12 well-trained male runners completed 4 running time to exhaustion (TTE) trials at a speed equivalent to 70% of VO2peak in a thermoneutral condition. Throughout each run, participants mouth rinsed and expectorated every 15 min either 25 mL of 6% CHO or a placebo (PLA) solution for 10 s. The four TTEs consisted of two trials in the euhydrated (EU-CHO and EU-PLA) and two trials in the dehydrated (DY-CHO and DY-PLA) state. Prior to each TTE run, participants were dehydrated via exercise and allowed a passive rest period during which they were fed and either rehydrated equivalent to their body mass deficit (i.e., EU trials) or ingested only 50 mL of water (DY trials).
RESULTS: CHO mouth rinsing significantly improved TTE performance in the DY compared to the EU trials (78.2 ± 4.3 vs. 76.9 ± 3.8 min, P = 0.02). The arousal level of the runners was significantly higher in the DY compared to the EU trials (P = 0.02). There was no significant difference among trials in heart rate, plasma glucose and lactate, and psychological measures.
CONCLUSIONS: CHO mouth rinsing enhanced running performance significantly more when participants were dehydrated vs. euhydrated due to the greater sensitivity of oral receptors related to thirst and central mediated activation. These results show that level of dehydration alters the effect of brain perception with presence of CHO.