Clonal growth of primary cultures of human hyaline chondrocytes in a defined medium

Rapid clonal growth of primary cultures of human costal chondrocytes in a defined medium has been achieved. The basal nutrient medium used for such growth is MCDB 104. It is prepared without linoleic acid and supplemented with 1 microgram/ml insulin, 100 ng/ml fibroblast growth factor, 1.0 x 10(-7) M dexamethasone, and 5 micrograms/ml mixed lipids, presented to the cells in the form of liposomes. The lipid supplement contains soybean lecithin, cholesterol, sphingomyelin, vitamin E, and vitamin E acetate. No expression of cartilage-like differentiation occurs in the defined medium. However, colonies grown for several days in the defined medium and then grown for an additional period of time in medium F12 supplemented with fetal bovine serum and chicken embryo extract synthesize large amounts of refractile matrix that is stained intensely by acidified alcian green, thus demonstrating that the cells growing in the defined medium are capable of expressing cartilage matrix in a permissive environment. Good clonal growth and expression of differentiation can also be obtained by inoculating primary cultures of human chondrocytes directly into the F12-serum-embryo extract medium.     

Evidence for a PGE-receptor in the rat kidney

A membrane preparation derived from homogenates of the rat kidney has been shown to possess a high affinity for prostaglandins of the E-series. Other prostaglandins including PGI₂ had characteristic but significantly weaker binding properties. A 15-hydroxyprostaglandin dehydrogenase (PGDH) was found to be associated with the membrane fraction studied. However it was possible to distinguish between this and the “receptor” binding by kinetic studies and by the use of a new inhibitor highly specific for PGDH.     

Regional localization of human beta-casein gene (CSN2) to 4pter-q21.

Milk proteins are crucial for the development of all newborn mammals. Caseins that constitute the bulk of the protein in mammalian milk have been shown to be members of a multigene family in at least two species. They are among the most rapidly diverging groups of proteins, and their numbers vary widely among species. beta- and kappa-Caseins are the only caseins present in human milk. Using polymerase chain reaction on genomic DNA from somatic cell hybrids, we have localized the human beta-casein gene (CSN2) to 4pter----q21.     

Stress-activated protein kinases are negatively regulated by cell density.

Stimulation by UV irradiation, TNFalpha, as well as PDGF or EGF activates the JNK/SAPK signalling pathway in mouse fibroblasts. This results in the phosphorylation of the N-terminal domain of c-Jun, increasing its transactivation potency. Using an antibody that specifically recognizes c-Jun phosphorylated at Ser63, we show that culture confluency drastically inhibited c-Jun N-terminal phosphorylation due to the inhibition of the JNK/SAPK pathway. Transfection experiments demonstrate that the inhibition occurs at the same level as, or upstream of, the small G-proteins cdc42 and Rac1. In contrast, the classical MAPK pathway was insensitive to confluency. The inhibition of JNK/SAPK activation depended on the integrity of the actin microfilament network. These results were confirmed and extended in monolayer wounding experiments. After PDGF, EGF or UV stimulation, c-Jun was predominantly phosphorylated in cells bordering the wound, which are the cells that move to occupy the wounded area. Thus, modulation of the stress-dependent signal cascade by confluency will restrict c-Jun N-terminal phosphorylation in response to mitogenic or chemotactic agents to cells that border a wounded area.     

Commentary on Coram et al. (2021) on the use of Facebook to understand marine mammal stranding issues in Southeast Asia

We reviewed Coram et al. (Biodivers Conserv 30:2341–2359, 2021, https://doi.org/10.1007/s10531-021-02196-6), a paper that highlights the use of social media data to understand marine litter and marine mammals in Southeast Asia. While we commend its intent, we find that the methodology used and conclusions drawn portray an incomplete and inaccurate perception of how strandings, stranding response, and analysis of stranding data have been conducted in the region. By focusing on investigative results revealed by a very limited search of one social media platform (Facebook), using only English keywords, and insufficient ground-truthing, Coram et al. (2021) have, unintentionally, given the perception that Southeast Asian scientists have not conducted even the bare minimum of investigation required to better understand the issue of marine litter and its impact on marine mammals. In this commentary we provide a more accurate account of strandings research in Asia and include recommendations to improve future studies using social media to assess conservation issues.