A standardized method to study immune responses using porcine whole blood

Background@#Peripheral blood mononuclear cells (PBMCs) are commonly used to assess in vitro immune responses. However, PBMC isolation is a time-consuming procedure, introduces technical variability, and requires a relatively large volume of blood. By contrast, whole blood assay (WBA) is faster, cheaper, maintains more physiological conditions, and requires less sample volume, laboratory training, and equipment. @*Objectives@#Herein, this study aimed to develop a porcine WBA for in vitro evaluation of immune responses. @*Methods@#Heparinized whole blood (WB) was diluted (non-diluted, 1/2, 1/8, and 1/16) in RPMI-1640 media, followed by phorbol myristate acetate and ionomycin. After 24 h, cells were stained for interferon (IFN)-γ secreting T-cells followed by flow cytometry, and the supernatant was analyzed for tumor necrosis factor (TNF)-α. In addition, diluted WB was stimulated by lipopolysaccharide (LPS) and polyinosinic:polycytidylic acid (poly I:C), reference strain KCTC3557 (RS), field isolate (FI), of heat-killed (HK) Streptococcus suis, and porcine reproductive and respiratory syndrome virus (PRRSV). @*Results@#The frequency of IFN-γ + CD3 + T-cells and concentration of TNF-α in the supernatant of WB increased with increasing dilution factor and were optimal at 1/8. WB TNF-α and interleukin (IL)-10 cytokine levels increased significantly following stimulation with LPS or poly I:C. Further, FI and RS induced IL-10 production in WB. Additionally, PRRSV strains increased the frequency of IFN-γ + CD4 - CD8 + cells, and IFN-γ was non-significantly induced in the supernatant of re-stimulated samples. @*Conclusions@#We propose that the WBA is a rapid, reliable, and simple method to evaluate immune responses and WB should be diluted to trigger immune cells.

Evaluation of metal contamination and phytoremediation potential of aquatic macrophytes of East Kolkata Wetlands, India

OBJECTIVES: The present study analyzes metal contamination in sediment of the East Kolkata Wetlands, a Ramsar site, which is receiving a huge amount of domestic and industrial wastewater from surrounding areas. The subsequent uptake and accumulation of metals in different macrophytes are also examined in regard to their phytoremediation potential. METHODS: Metals like cadmium (Cd), copper (Cu), manganese (Mn), and lead (Pb) were estimated in sediment, water and different parts of the macrophytes Colocasia esculenta and Scirpus articulatus. RESULTS: The concentration of metals in sediment were, from highest to lowest, Mn (205.0±65.5 mg/kg)>Cu (29.9±10.2 mg/kg)>Pb (22.7±10.3 mg/kg)>Cd (3.7±2.2 mg/kg). The phytoaccumulation tendency of these metals showed similar trends in both native aquatic macrophyte species. The rate of accumulation of metals in roots was higher than in shoots. There were strong positive correlations (p<0.001) between soil organic carbon (OC) percentage and Mn (r =0.771), and sediment OC percentage and Pb (r=0.832). Cation exchange capacity (CEC) also showed a positive correlation (p<0.001) with Cu (r=0.721), Mn (r=0.713), and Pb (r=0.788), while correlations between sediment OC percentage and Cu (r=0.628), sediment OC percentage and Cd (r=0.559), and CEC and Cd (r=0.625) were significant at the p<0.05 level. CONCLUSIONS: Bioaccumulation factor and translocation factors of these two plants revealed that S. articulatus was comparatively more efficient for phytoremediation, whereas phytostabilization potential was higher in C. esculenta.

Antidiarrhoeal and neuropharmacological activities of Avicennia officinalis Linn.

The methanolic extract of the leaves of Avicennia officinalis Linn. was tested for its antidiarrhoeal and neuropharmacological activities The extract showed a significant antidiarrhoeal activity against castor oil induced diarhoea in mice in which it decreased the frequency of defecation and increased mean latent period at the dose of 500 mg/kg body weight Moreover, it potentiated the pentobarbital induced sleeping time in mice at the dose of 500 mg/kg body weight, and decreased the number of holes crossed from one chamber in the hole cross test and decreased the head dip responses. The overall results tend to suggest the antidiarrhoeal and central nervous system depressant activities of the extract