Diepoxybutane (DEB) is the most potent active metabolite of butadiene, a regulated air pollutant. We previously reported the occurrence of DEB-induced, p53-dependent, mitochondrial-mediated apoptosis in human lymphoblasts. The present study investigated the role of the extracellular signal-regulated protein kinases 1 and 2 (ERK1/2) pathway in DEB-induced apoptotic signaling in exposed human lymphoblasts. Activated ERK1/2 and mitogen-activated protein (MAP) kinase/ERK1/2 kinase (MEK) levels were significantly upregulated in DEB-exposed human lymphoblasts. The MEK inhibitor PD98059 and ERK1/2 siRNA significantly inhibited apoptosis, ERK1/2 activation, as well as p53 and phospho-p53 (serine-15) levels in human lymphoblasts undergoing DEB-induced apoptosis. Collectively, these results demonstrate that DEB induces apoptotic signaling through the MEK-ERK1/2-p53 pathway in human lymphoblasts. This is the first report implicating the activation of the ERK1/2 pathway and its subsequent role in mediating DEB-induced apoptotic signaling in human lymphoblasts. These findings contribute towards the understanding of DEB toxicity, as well as the signaling pathways mediating DEB-induced apoptosis in human lymphoblasts.

Abstract:Galectins constitute an evolutionarily conserved family of β-galactoside-binding proteins. 3). Beta-actin and GAPDH housekeeping genes were used as internal controls. Fold changes in 22 transcript abundance were compared to non-pregnant goats and calculated using the Livak method. 23Secretion of GALS-1, -3 and -9 in plasma was detected using ELISA. Data were analyzed using SAS

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Galectins (GAL) constitute an evolutionarily conserved family of β-galactoside-binding proteins that are secreted. They are involved in the regulation of homeostasis, innate and adaptive immune responses to infectious challenge. The mushroom Coriolus versicolor (CV) has been reported to boost suppressed immune function, extending the survival rate and improving quality of life in man. The objective of this study was to evaluate the effect of mushroom probiotics on the secretion of Galectins in goat blood. Ten BoerXSpanish female goats (5 weeks of age) were used for this study. Powdered CV was soaked in cold water and sterile filtered. Following initial screening for infection, goats were assigned to two groups of five (n = 10). Goats were drenched daily with 10 mL cold (treatment I) extract for 4 weeks. A control group of five age-matched goats received sterile water (treatment II). Blood samples were collected on a weekly basis. The concentration plasma protein was determined by using Pierce BCA kit (Thermo Scientific Pierce, Rockford, IL). Galectins-1, -3, -8, -9 and -12 concentration was detected by using a commercial ELISA kits (ABclonal Biotechnology, Woburn, MA). There was an effect of treatment in total plasma protein concentration when compared to the control group (P < 0.02). Galectins tested were secreted in both control and treatment groups. Treatment with CV decreased the concentration of Gal-1, 8 and 9 and increased the concentration of Gal-3 and -12 (P < 0.05). Results from this study indicate that mushroom probiotics can modulate the secretion of GAL.

The objective of this study was to assess the expression and secretion of Galectins (Gal) in cow milk. Cow milk contains a range of proteins of moderate or low abundance that contribute to host defense. However, host defense proteins in milk were not fully discovered. Galectins belong to the lectin family that recognizes specific carbohydrates on cells and involved in innate immune responses. Holstein Friesian cows (n = 16) from North Carolina A&T diary unit were used for this study. Based on the Dairy Heard farm index, eight cows each were assigned to the high or low SCC group. Total RNA was isolated from somatic cells converted to cDNA, for real-time PCR. Cow-specific Gal primers were designed using the NCBI Primer 3 tool. Housekeeping genes RPLP0, UCHL5, and beta-actin were served as internal controls. Total whey protein concentration was determined using a BCA kit. Secretion of Gal was assessed using a specific ELISA kit. Data were analyzed using the Proc ANOVA procedure in SAS 9.4. Galectin was transcribed and secreted in milk. Transcription of Galectin was different in both HSCC and LSCC groups. Total protein concentration remained the same in both groups. Secretion of galectins was different between the HSCC and LSCC group but not significantly. The observed difference in HSCC and LSCC cows warrants further study using more animals; this will aid in a better definition of the role of Gal in the milk host defense. These may also aid as the diagnostic biomarkers for certain infections.