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The cDNAs for cytochrome P450 monooxygenase (designated as CYP97C27 by D. Nelson's group) and NADPH-cytochrome P450 reductase (designated as CPR I based on its classification) were isolated from Croton stellatopilosus leaves, which actively biosynthesise plaunotol (18-OH geranylgeraniol). CYP97C27 and CPR I contain open reading frames encoding proteins of 471 and 711 amino acids with predicted molecular masses of 53 and 79kDa, respectively. By aligning the deduced sequences of CYP97C27 and CPR I with other plant species, all functional domains of CYP97C27 (heme and oxygen binding) and CPR I (CYP- and FMN, FAD, and NADPH cofactor binding) were identified. Amino acid sequence comparison indicated that both CYP97C27 (85-93%) and CPR I (79-83%) share high sequence identities with homologous proteins in other plant species, suggesting that CYP97C27 belongs to the CYP97C subfamily and that CPR I belongs to class I of the dicotyledonous CPR. Functional characterisation of both enzymes, produced in Escherichia coli (pET32a/BL21(DE3)) as recombinant proteins, showed that simultaneous incubation of CYP97C27 and CPR I with the substrate geranylgeraniol (GGOH) and coenzyme NADPH led to formation of the product plaunotol. In C. stellatopilosus, the levels of the CYP97C27 and CPR I transcripts were highly correlated with those of several mRNAs involved in the plaunotol biosynthetic pathway, suggesting that CYP97C27 and CPR I are the enzymes that catalyse the last hydroxylation step of the pathway.

An entomological index is widely used for determining dengue transmission and mosquito populations. In this study, we determined the association between Aedes albopictus mosquito counts in three areas with different topographical features from six districts in Nakhon Si Thammarat and infected patients reported by the Ministry of Public Health's Department of Disease Control, using entomological indices, such as the sticky pad index and the sweep net index. The sampling region was selected from the top six districts of the province based on the number of reported dengue cases per 100,000 people. The regions were Khanom, Mueang, Pak Phanang, Phrom Khiri, Thung Song, and Thung Yai. They were divided into three categories of geographical features, including city, beach, and mountain. The mosquitos were collected by walking in a star form inside a 5 m diameter circle. The mosquitoes were separated into groups based on their species, and the percentile of the index was computed. We found that the number of mosquitoes was the highest in Pak Phanang and lowest in Khanom. The two indices and the number of A. albopictus collected from the six sampling locations were not significantly different (t = 1.31, p > 0.05). The coastline region had the highest number of mosquitoes and was substantially different from the other two regions (cosmopolitan and mountain) (F = 4.30; p < 0.05). By performing Pearson's correlation analysis, we found a negative correlation between dengue cases and the entomological index (r = -0.472; N = 36). Similarly, the number of patients showed a negative correlation with relative humidity (r = -0.355) but a positive correlation with temperature (r = 0.064). The epidemic pattern shifted throughout the year, and the number of cases then increased 2-4 weeks after the insect was found.

TLC (thin-layer chromatography) is a standard technique for simultaneously screening and monitoring chemical character in a large number of samples. The goal of this study was to explore if alpha-mangostin (a common secondary metabolite) could be detected in mangosteen pericarp phytochemical fingerprints and how much of it could be represented using a standard calibration curve. The antioxidant activity has been touted as a primary active ingredient in many commercial goods, including cosmetics and food supplements. To test for the antioxidant reaction to the DPPH onto the TLC plate, mangosteen pericarps were obtained from 12 orchards that were grown without pesticides in Cha-Uat (CU), Lan Saka (LS), and Phrom Khiri (PK) districts of Nakhon Si Thammarat. The samples were dried and powdered before being extracted with methanol using the reflux technique. After that, the TLC was utilized to determine the phytochemical fingerprint. The quality of phytochemicals from the LS orchards was found to differ from CU and PK samples, with different bands of compounds at RF 0.2, 0.25, and 0.57. The average amount of alpha-mangostin in the 12 samples was not statistically different, according to a one-way analysis of variance with a p-value of 0.05. The average range of interest (ROI) intensity area of the antioxidant was investigated using a one-way analysis of variance with a p-value of 0.05 and repeated comparisons across the sample groups by Tukeys multiple comparison test. The average antioxidant reaction between the CU and PS group and the CU and PK group was significantly different.

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