Sequence analysis of new variants of porcine epidemic diarrhea virus in Luzon, Philippines, in 2017.

Porcine epidemic diarrhea virus (PEDV) is a coronavirus that causes emaciation and watery diarrhea in pigs. First identified in Europe in 1977, it eventually spread to Asia and North America, causing deadly outbreaks in neonatal piglets. In the Philippines, PEDV has caused several recorded outbreaks since 2005. However, DNA sequencing studies of local PEDV strains remain few and are limited to gene and gene fragment sequencing. Therefore, to provide updated sequence information about recent PEDV strains in the country, we performed reverse transcription PCR and sequencing of PEDV from swab samples collected from swine farms in the Philippines in 2017. Here, we report the first published whole genome sequence of PEDV from the Philippines as well as CO-26K equivalent (COE) domain sequences of strains from three provinces in Luzon where PEDV was detected in 2017. Sequence analysis suggested that PEDV from both the classical (genotype 1) and pandemic (genotype 2) groups are present in the Philippines, with possible East Asian and North American origins.

Heat Shock Protein Augmentation of Angelica gigas Nakai Root Hot Water Extract on Adipogenic Differentiation in Murine 3T3-L1 Preadipocytes.

There is a high association of heat shock on the alteration of energy and lipid metabolism. The alterations associated with thermal stress are composed of gene expression changes and adaptation through biochemical responses. Previous study showed that Angelica gigas Nakai (AGN) root extract promoted adipogenic differentiation in murine 3T3-L1 preadipocytes under the normal temperature condition. However, its effect in heat shocked 3T3-L1 cells has not been established. In this study, we investigated the effect of AGN root hot water extract in the adipogenic differentiation of murine 3T3-L1 preadipocytes following heat shock and its possible mechanism of action. Thermal stress procedure was executed within the same stage of preadipocyte confluence (G0) through incubation at 42°C for one hour and then allowed to recover at normal incubation temperature of 37°C for another hour before AGN treatment for both cell viability assay and Oil Red O. Cell viability assay showed that AGN was able to dose dependently (0 to 400 µg/mL) increase cell proliferation under normal incubation temperature and also was able to prevent cytotoxicity due to heat shock accompanied by cell proliferation. Confluent preadipocytes were subjected into heat shock procedure, recovery and then AGN treatment prior to stimulation with the differentiation solution. Heat shocked preadipocytes exhibited reduced differentiation as supported by decreased amount of lipid accumulation in Oil Red O staining and triglyceride measurement. However, those heat shocked preadipocytes that then were given AGN extract showed a dose dependent increase in lipid accumulation as shown by both evaluation procedures. In line with these results, real-time polymerase chain reaction (RT-PCR) and Western blot analysis showed that AGN increased adipogenic differentiation by upregulating heat shock protection related genes and proteins together with the adipogenic markers. These findings imply the potential of AGN in heat shock amelioration among 3T3-L1 preadipocytes through heat shock factor and proteins augmentation and enhanced adipogenic marker expression.