Development of flow-injection liposome immunoanalysis (FILIA) for imazethapyr

Imazethapyr is the herbicide developed for use in leguminous crops. In this study, flow-injection liposome immunoanalysis (FILIA) has been shown to be capable of measuring imazethapyr in a buffered solution with a detection limit of 0.1 ppb through the optimization process. Protein A coated glass beads covalently conjugated with antibody were contained in a glass column, and this column was used as an immunoreactor. Liposomes which encapsulated a fluorescent dye, sulforhodamine B (SRB) or carboxyfluorescein (CF), generated the analytical signal. By loading larger volumes of sample onto the column, it was shown that the detection limit could be lowered. Liposomes containing carboxyfluorescein gave more sensitive response and a lower detection limit than those with sulforhodamine B. Also, improved response was obtained by using a smaller flow cell in the fluorescence detector due to the reduced dilution effect.     

Orthogonal array design (OAD) for the optimization of mercury extraction from soils by dilute acid with microwave heating

The use of dilute acid for mercury extraction from geological samples with microwave assisted heating was investigated. An orthogonal array design OA(16)(4(1)x2(12)) was applied to select the optimum conditions. The effects of the concentration of HNO(3), the additions of HCl and H(2)S0(4), the extraction time and their interactions were evaluated by the mixed-level orthogonal array design (OAD). The results indicated that the addition of small amount of HCl would improve significantly the extraction of mercury from soil matrices. From the study, it is seen that the mercury in the soil matrices could be extracted completely by 14% (v/v) HNO(3) with small quantity of HCl. The effects of the pressure within the digestion vessel and the sample mass were studied under the optimum conditions derived from the OAD. The extraction methods with optimum conditions were evaluated by determining mercury in NIST SRM 1645 River Sediment, NIES CRM No. 2 Pond Sediment and NRCC BCSS-1 Marine Sediment. Recoveries of 78-109% were achieved. Good mean recoveries of 95-98% were also obtained from real soil samples spiked with different species of mercury.     

Gaucher disease-associated glucocerebrosidases show mutation-dependent chemical chaperoning profiles

Gaucher disease is a lysosomal storage disorder caused by deficient glucocerebrosidase activity. We have previously shown that the cellular activity of the most common Gaucher disease-associated glucocerebrosidase variant, N370S, is increased when patient-derived cells are cultured with the chemical chaperone N-nonyl-deoxynojirimycin. Chemical chaperones stabilize proteins against misfolding, enabling their trafficking from the endoplasmic reticulum. Herein, the generality of this therapeutic strategy is evaluated with other glucocerebrosidase variants and with additional candidate chemical chaperones. Improved chemical chaperones are identified for N370S glucocerebrosidase. Moreover, we demonstrate that G202R, a glucocerebrosidase variant that is known to be retained in the endoplasmic reticulum, is also amenable to chemical chaperoning. The L444P variant is not chaperoned by any of the active site-directed molecules tested, likely because this mutation destabilizes a domain distinct from the catalytic domain.