The mammalian liver possesses a remarkable regenerative ability. Two modes of damage response have been described: (1) The ''oval cell'' response emanates from the biliary tree when all hepatocytes are affected by chronic liver disease. (2) A massive, proliferative response of mature hepatocytes occurs upon acute liver damage such as partial hepatectomy (PHx). While the oval cell response has been captured in vitro by growing organoids from cholangiocytes, the hepatocyte proliferative response has not been recapitulated in culture. Here, we describe the establishment of a long-term 3D organoid culture system for mouse and human primary hepatocytes. Organoids can be established from single hepatocytes and grown for multiple months, while retaining key morphological, functional and gene expression features. Transcriptional profiles of the organoids resemble those of proliferating hepatocytes after PHx. Human hepatocyte organoids proliferate extensively after engraftment into mice and thus recapitulate the proliferative damage-response of hepatocytes.
Brassinolides (BRs) are naturally occurring substances, which modulate plant growth and development events and have been known to improve the crop tolerance to abiotic stresses. In this study, possible role of exogenously applied brassinolide (BR) in alleviating the detrimental effects of drought in maize was evaluated in a rain‐protected wire‐house. Maize was subjected to drought at the start of tasseling for 6 days by withholding water application followed by foliar spray of BR (0.1 mg l−1) to assess the changes in growth, gas exchange, chlorophyll contents, protein, relative leaf water contents (RLWC), proline, malonialdehyde (MDA) and enzymatic antioxidants. Drought substantially reduced the maize growth in terms of plant height, leaf area and plant biomass. Moreover, substantial decrease in gas exchange attributes (net photosynthetic rate (A), transpiration rate (E), stomatal conductance (gs), water use efficiency (WUE), instantaneous water use efficiency (WUEi) and intercellular CO2 (Ci) was also recorded. However, exogenous application of BR remarkably improved the gas exchange attributes, plant height, leaf area, cobs per plant, seedling dry weight both under drought and well‐watered conditions. BR‐induced promotion in growth and physiological and metabolic activities were mediated through increased protein synthesis enabling maintenance of tissue water potential and activities of antioxidant enzymes lowering the lipid peroxidation under drought.
Bleeding disorders and thrombotic complications constitute a major cause of death and disability worldwide. While it’s known that the complement and coagulation systems interact, no studies have investigated the specific role or mechanisms of lectin-mediated coagulation in vivo. Ferric chloride (FeCl3) treatment resulted in intra-arterial occlusive thrombogenesis within 10min in wild-type (WT) and C2/fB null mice. In contrast, MBL null and MASP-1/-3 KO mice had significantly decreased FeCl3-induced thrombogenesis. Reconstitution with rhMBL restored FeCl3-induced thrombogenesis in MBL null mice to levels comparable to WT mice, suggesting a significant role of the MBL-MASP complex for in vivo coagulation. Additionally, whole blood aggregation demonstrated increased MBL-MASP complex-dependent platelet aggregation. In vitro, MBL-MASP complexes were captured on mannan-coated plates and cleavage of a chromogenic thrombin substrate (S2238) was measured. We observed no significant differences in S2238 cleavage between WT, C2/fB null, MBL-A−/− or MBL-C−/− sera, however MBL null or MASP-1/-3 KO mouse sera demonstrated significantly decreased S2238 cleavage. Recombinant human (rh)MBL alone failed to cleave S2238, however cleavage was restored when rMASP-1 was added to either MASP-1/-3 KO sera or rhMBL. Taken together, these findings indicate that MBL-MASP complexes, and specifically MASP-1, play a key role in thrombus formation in vitro and in vivo.
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