MLK3 is a novel target of dehydroglyasperin D for the reduction in UVB‐induced COX‐2 expression in vitro and in vivo

Dehydroglyasperin D (DHGA‐D), a compound present in licorice, has been found to exhibit anti‐obesity, antioxidant and anti‐aldose reductase effects. However, the direct molecular mechanism and molecular targets of DHGA‐D during skin inflammation remain unknown. In the present study, we investigated the effect of DHGA‐D on inflammation and its mechanism of action on UVB‐induced skin inflammation in HaCaT human keratinocytes and SKH‐1 hairless mice. DHGA‐D treatment strongly suppressed UVB‐induced COX‐2 expression, PGE2 generation and AP‐1 transactivity in HaCaT cells without affecting cell viability. DHGA‐D also inhibited phosphorylation of the mitogen‐activated protein kinase kinase (MKK) 3/6/p38, MAPK/Elk‐1, MKK4/c‐Jun N‐terminal kinase (JNK) 1/2/c‐Jun/mitogen, and stress‐activated protein kinase (MSK), whereas phosphorylation of the mixed‐lineage kinase (MLK) 3 remained unaffected. Kinase and co‐precipitation assays with DHGA‐D Sepharose 4B beads showed that DHGA‐D significantly suppressed MLK3 activity through direct binding to MLK3. Knockdown of MLK3 suppressed COX‐2 expression as well as phosphorylation of MKK4/p38 and MKK3/6/JNK1/2 in HaCaT cells. Furthermore, Western blot assay and immunohistochemistry results showed that DHGA‐D pre‐treatment significantly inhibits UVB‐induced COX‐2 expression in vivo. Taken together, these results indicate that DHGA‐D may be a promising anti‐inflammatory agent that mediates suppression of both COX‐2 expression and the MLK3 signalling pathway through direct binding and inhibition of MLK3.     

Two-stage anaerobic treatment of dairy wastewater using HUASB with PUF and PVC carrier

In the present study, an attempt has been made to treat dairy wastewater entirely via anaerobic treatment over a period of 215 days, using two-stage Hybrid Upflow Anaerobic Sludge Blanket (HUASB) reactors, which offer the advantages associated both with fixed film and upflow sludge blanket treatments. A HUASB with polyurethane foam cubes was used for stage I, and a HUASB utilizing PVC-cut rings was used for stage II. The output from stage I was used as the input for stage II. The two-stage reactor was operated at an organic loading rate that varied from 10.7 to 21.4 kg COD m³/d for a period of 215 days, including the start-up period. The ideal organic loading rate for the two-stage reactor was 19.2 kg COD/m³/d. A further 21.4 kg COD m³/d increase in the organic loading rate resulted in the souring of the reactor function in stage I, which consequently reduced the overall reactor performance. Combined COD removal during the stable operation period (10.7 to 19.2 kg COD m³/d) occurred in a range between 97 and 99%. The methane content in the biogas varied from 65 to 70% in stage I, and from 63 to 66% in stage II. The two-stage anaerobic treatment using HUASB with PUF and PVC described in this work is expected to constitute a better alternative for the complete treatment of dairy wastewater than high-rate anaerobic, anaerobic/aerobic, and two-phase anaerobic treatment methods.