Species-diverse coral communities on an artificial substrate at a tuna farm in Amami, Japan.

Tuna-farming is expanding worldwide, necessitating the monitoring/managing of its effects on the natural environment. In Japan, tuna-farming is conducted on coral reefs that have been damaged by mass-bleaching events and crown-of-thorns starfish (COTS) outbreaks. This study focused on the coral community on an artificial substrate of tuna-farm to reveal the possible effects of tuna-farming on the natural environment.

Determination of plasma thiol bound to albumin using affinity chromatography and high-performance liquid chromatography with fluorescence detection: ratio of cysteinyl albumin as a possible biomarker of oxidative stress.

We examined the influence of oxidative stress on the relative amounts of various albumin-bound thiols in human plasma. To determine the ratio of thiols existing as mixed disulfides following oxidation, we developed a method combining fast purification of albumin using affinity columns and high-performance liquid chromatography (HPLC) with fluorescence detection for low molecular weight thiols which were labeled after reduction. When the effect of exposure of plasma to radical oxygen species on binding of thiols to albumin was determined by the present method, significant increases in the ratio of cysteine bound to albumin (Alb-Cys) to total cysteine were clearly demonstrated.

Measurement of homocysteine thiolactone hydrolase activity using high-performance liquid chromatography with fluorescence detection and polymorphisms of paraoxonase in normal human serum.

We developed a non-radioactive and sensitive assay method for measurement of the HTL hydrolase (HTLase) activity in biological samples, using OPA as a fluorescent post-labeling agent, l-homocysteine thiolactone (L-HTL) as the substrate, and HPLC to achieve rapid and selective separation of the substrate and product. The method was applied to measure the activity of HTLase in human, rabbit, rat and mouse serum samples. In addition, the correlation between the serum HTLase activity and PON1 polymorphisms in Japanese subjects was also investigated.

Determination of homocysteine thiolactone and homocysteine in cell cultures using high-performance liquid chromatography with fluorescence detection.

A sensitive and simple method utilising fluorometric detection for the simultaneous routine monitoring of homocysteine thiolactone (HTL) and homocysteine (Hcy) in biological samples has been developed. Separation relies on isocratic ion-pairing and reversed-phase chromatography while the principle of the detection is that the lactone ring in HTL molecule is cleaved with an alkali to produce Hcy, which reacts with ortho-phthalaldehyde (OPA) in the absence of an added thiol reagent to form a stable fluorescent derivative. The method has a sensitivity of 200 fmol of HTL and 100 fmol for Hcy in the sample.