MATERIALS AND METHODS: Thus, crab samples for this study were sampled from the wild habitat at Setiu wetlands, Terengganu, Malaysia. A total of 100 samples of with the same size (80±5 mm carapace width) were sampled and the eyestalk dissected for LF extraction. The determination of LF sites and levels in the eyestalks organ was taken from the area between the distal tangential layer (DTL) and proximal tangential layer (PTL). The lower part of the eyestalk was taken from the PTL until the end of the eyestalk.
RESULTS: The upper part of the crab's eyestalk was higher in the males crabs compared to the females crabs. LF index also shown that the upper part of crab's eyestalk have higher concentration compared to the lower part.
CONCLUSION: The left crab's eyestalk had the higher LF index especially in males compared to females but the total concentration was higher in female crabs. Knowing which part has highly dense accumulation of LF helps in LF detection of the tissue and further helps for age determination for this species.
METHOD: Lifespan was determined by counting the number of surviving nematodes daily under a dissecting microscope after treatment with hydrogen peroxide and the tocotrienol-rich fraction. The evaluated oxidative markers included lipofuscin, which was measured using a fluorescent microscope, and protein carbonyl and 8-hydroxy-2'-deoxyguanosine, which were measured using commercially available kits.
RESULTS: Hydrogen peroxide-induced oxidative stress significantly decreased the mean lifespan of C. elegans, which was restored to that of the control by the tocotrienol-rich fraction when administered before or both before and after the hydrogen peroxide. The accumulation of the age marker lipofuscin, which increased with hydrogen peroxide exposure, was decreased with upon treatment with the tocotrienol-rich fraction (p<0.05). The level of 8-hydroxy-2'-deoxyguanosine significantly increased in the hydrogen peroxide-induced group relative to the control. Treatment with the tocotrienol-rich fraction before or after hydrogen peroxide induction also increased the level of 8-hydroxy-2'-deoxyguanosine relative to the control. However, neither hydrogen peroxide nor the tocotrienol-rich fraction treatment affected the protein carbonyl content of the nematodes.
CONCLUSION: The tocotrienol-rich fraction restored the lifespan of oxidative stress-induced C. elegans and reduced the accumulation of lipofuscin but did not affect protein damage. In addition, DNA oxidation was increased.