Mars J et al. (2024) MicroPubl Biol "Low-cost, rapid and reliable DNA extraction from mammalian organoids using a C. elegans toolbox."

Mars J, Galli M, Darmasaputra GS, Korswagen HC

[MicroPubl Biol, 2024]

High-quality DNA extraction from organoids is an important step in molecular genetics research. Here, we show that a lysis buffer from the field of <i>Caenorhabditis elegans</i> research, called Single Worm Lysis Buffer (SWLB), is a low-cost, yet reliable method for DNA extraction from mammalian organoids. SWLB is superior in terms of price, storage, hands-on time and sustainability compared to current standardized DNA extraction protocols, while equally effective. This work indicates that it is useful to compare methods from different model systems, such as mammalian organoids and invertebrate nematodes, to find useful alternatives for research methodologies.

Filippidis, G et al. (2005) J Phys D Appl Phys "Imaging of Caenorhabditis elegans samples and sub-cellular localization of new generation photosensitizers for photodynamic therapy, using non-linear microscopy."

Filippidis, G, Papazoglou, TG, Voglis, G, Kapsokalyvas, D, Tavernarakis, N, Kouloumentas, C

[J Phys D Appl Phys, 2005]

Two-photon excitation fluorescence (TPEF) and second-harmonic generation (SHG) are relatively new promising tools for the imaging and mapping of biological structures and processes at the microscopic level. The combination of the two image-contrast modes in a single instrument can provide unique and complementary information concerning the structure and the function of tissues and individual cells. The extended application of this novel, innovative technique by the biological community is limited due to the high price of commercial multiphoton microscopes. In this study, a compact, inexpensive and reliable setup utilizing ferntosecond pulses for excitation was developed for the TPEF and SHG imaging of biological samples. Specific cell types of the nematode Caenorhabditis elegans were imaged. Detection of the endogenous structural proteins of the worm, which are responsible for observation of SHG signals, was achieved. Additionally, the binding of different photosensitizers in the HL-60 cell line was investigated, using non-linear microscopy. The sub-cellular localization of photosensitizers of a new generation, very promising for photodynamic therapy (PDT), (Hypericum perforatum L. extracts) was achieved. The sub-cellular localization of these novel photosensitizers was linked with their photodynamic action during PDT, and the possible mechanisms for cell killing have been elucidated.

Morrison, Kayleigh N. et al. (2021) MicroPubl Biol

Morrison, Kayleigh N., Ragle, James Matthew, Shakes, Diane C., Uyehara, Christopher M., Ward, Jordan D.

[MicroPubl Biol, 2021]

During the process of cell differentiation, specific cytoskeletal proteins can sequentially assemble into a wide variety of diverse molecular superstructures. Nematode spermatogenesis provides a powerful system for studying these transitions since sperm-specific transcription ceases prior to the meiotic divisions and translation ceases shortly thereafter (Chu and Shakes, 2013). Therefore, structural transitions that follow the meiotic divisions must be carried out by the remodeling of already synthesized proteins. The Major Sperm Protein (MSP) is a nematode-specific cytoskeletal element whose polymerization dynamics drive the pseudopod-based motility of the activated sperm (Roberts, 2005). In C. elegans, MSP additionally functions as the extracellular signaling molecule for triggering both ovulation and oocyte maturation (Miller et al., 2003). MSP is highly abundant in sperm, where it reaches 10-15% of total and 40% of soluble cellular protein (Roberts 2005). Within developing spermatocytes, MSP is packaged into fibrous bodymembranous organelle (FB-MO) complexes (Fig. 1A, Roberts et al., 1986). By assembling into paracrystalline FBs, MSP is both sequestered away from the critical meiotic processes of chromosome segregation and cytokinesis while also being packaged for efficient segregation into spermatids during the post-meiotic partitioning process (Chu and Shakes 2013, Nishimura and LHernault, 2010, Price et al., 2021). Following the meiotic divisions and sperm individualization, FBs disassemble, and MSP disperses as dimers throughout the spermatid cytoplasm (Fig. 1A). When sperm activate to form motile spermatozoa, MSP polymerization within the pseudopod drives the motility of the crawling sperm (Chu and Shakes, 2013). Thus, MSP exists in at least three distinct molecular states: 1) in highly organized paracrystalline FBs within developing spermatocytes 2) as unpolymerized dimers within spermatids, and 3) in dynamically polymerizing filaments and fibers within crawling spermatozoa.

Xue J et al. (2020) J Food Biochem "The Sirtuin-1 relied antioxidant and antiaging activity of 5,5'-diferulic acid glucoside esters derived from corn bran by enzymatic method."

Xue J, Zhang G, Sheng X, Zhang BJ, Zhang C

[J Food Biochem, 2020]

Maize is the food crop with the highest total output in the world. However, corn bran is only a by-product with low price. The 5,5'-diferulic acid glucoside esters (DFG) were obtained from corn bran using the enzymatic method. DFG showed obvious antioxidant capacity in cell, Caenorhabditis elegans (C. elegans) and in mouse. DFG decreased ROS and MDA content in 500M H₂ O₂ stimulated ARPE-19 cells to 48.6% and 32.2%, respectively. DFG decreased ROS content in C. elegans to 49.1% and MDA content in acute ethanol (50%, 12ml/kg) stimulated mouse to 30.4%. DFG also increased SOD protein content significantly in cell, C. elegans and mouse to 175.5%, 120.1%, and 126.2%, respectively. DFG significantly extended the lifespan of C. elegans both under heat stress and natural situation. The median survival time was prolonged to 133.3% and 116.7%, respectively. This capacity relied on the SIR-2.1 activity. SIR-2.1 is an ortholog of human Sirtuin-1 (SIRT-1). DFG also upregulated SIRT-1 and PCG-1 expression level obviously in H₂ O₂ -stimulated ARPE-19 cells (to 134.4% and 127.1%, respectively) and in acute ethanol stimulated mouse eyes (to 135.1% and 111.5%, respectively) and liver (to 123.3% and 113.6%, respectively). These results indicate that DFG has multiple bioactivities. Our research provides a new application prospect of corn bran. And to our best knowledge, this is the first time, the sirtuins-relied lifespan extension activity of the 5,5'-diferulic acid extracted from corn bran was reported. PRACTICAL APPLICATIONS: The traditional method for extracting diferulic acid from corn bran is to use the strong alkali. Obviously, this is not welcomed by the food industry. We employed the biological enzyme method in a relatively mild pH range during the extraction process. It is more environmentally friendly and more economical. DFG can be added as a raw material for functional foods like yogurt, fruit juice, and cereals. As well, the solid precipitate obtained after extraction can also be used as high-quality dietary fiber to produce functional food. Meanwhile, concerning for the 5,5'-diferulic acid derived from corn bran, the relevant research is still not abundant. And to our best knowledge, we have reported for the first time about the effect of this kinds of diferulic acid on prolonging life span and its SIRT-1-dependent activity. It also provides a new perspective for the study of diferulic acid.