The b-HLH-LZ domain of c-Myc is a key target for the development of cancer therapies by blunting its binding to DNA with cell penetrant b-HLH-LZs and/or by stabilizing it into a state that cannot recognize Max to activate and amplify transcription of oncogenic genes. Although recent milestones have been reached with DNA binding blunting of c-Myc with the cell penetrant b-HLH-LZ Omomyc, the targeting of its b-HLH-LZ with small molecules, peptides, or proteins is lagging. As reviewed recently, the main problem relies in the intrinsically disordered nature of the b-HLH-LZ of c-Myc. This greatly complicates the classical approach of targeting a docking site with inhibitors. The solution state methods such as NMR are progressing towards the characterization of the ensembles of structures or states the b-HLH-LZ can adopt. However, the delicate balance that dictates the population of these dynamically interchanging states relies on its primary structure and the weak polar, electrostatic and hydrophobic interactions allowed. In this context, it is of the utmost importance to study the b-HLH-LZ of c-Myc in its WT background and avoid the use of tags such as His-tags. These tags could disrupt the balance of forces which could alter the conformational and physical transitions and states it can undergo and adopt. Here, we describe a robust protocol to express the WT b-HLH-LZ in E. coli and purify it, without the need of tags, to obtain the required quantities for solution state biophysical characterization such as NMR.The MYC transcription factor coordinates a wide range of intra- and extracellular processes associated with tissue proliferation and regeneration. While these processes are typically tightly regulated in physiological conditions, they become deregulated in cancer, where MYC is oncogenically activated.The last decade has seen MYC progress from a renowned undruggable target to a hot topic in the cancer therapy field, as proof emerged from mouse models that its inhibition constitutes an effective and broadly applicable approach to fight cancer. However, there are several aspects of MYC biology that still appear to be elusive and maintain the interest in further studying this intriguing protein. Since MYC's discovery, more than four decades ago, multiple strategies have been developed to study it, related to the many and varied facets of its biology. This new version of The Myc gene Methods and Protocols provides valuable tips from key "inhabitants of the MYC world," which significantly increase the reach of our investigative tools to shed light on the mysteries still surrounding MYC. This knowledge mobilization project was conceptualized to increase awareness among breastfeeding mothers and the general public on safe infant feeding practices during the COVID-19 pandemic by addressing myths and misconceptions associated with breastfeeding practices, guiding breastfeeding mothers to make informed decisions around child feeding practices, and offering meaningful guidance in simple language through a short online animated video. This project was undertaken in four phases. During phase 1, an informal discussion was held with the breastfeeding mothers, service providers, and community partner in identifying issues surrounding lactation counselling facilities during the COVID-19 pandemic. During phase 2, recommendations from 23 organizations with regard to breastfeeding during COVID-19 were reviewed and analyzed. PF-562271 purchase During phase 3, using evidence from reliable sources, a 5-minute animated e-resource on breastfeeding during COVID-19 was conceptualized and developed. During phase 4, the e-resourcprotecting breastfeeding during the COVID-19 pandemic. It further illustrated how informational e-resources can protect breastfeeding in situations where breastfeeding mothers' access to healthcare services is compromised. Neospora caninum, an obligate intracellular parasite of the phylum Apicomplexa, typically causes an illness known as neosporosis. Dense granule proteins (GRAs) are secreted by apicomplexan and constitute the parasitophorous vacuoles (PVs) structure where tachyzoites proliferate after invasion into host cells. In Toxoplasma gondii, TgGRA3 proteins are strongly associated with PVs membrane and enhance its virulence in vivo, however, research on NcGRA3 has not been reported. Here, a novel NcGRA3 protein in N. caninum was discovered using bioinformatics analysis; the location of NcGRA3 was determined in the extracellular Nc-1 tachyzoites and intracellular PVs after invasion using immunofluorescence assays; the NcGRA3 protein existing form in the PVs membranes was analyzed using western blotting. NcGRA3 shared 41.67% nucleotide homology and 22.17% amino acid homology with TgGRA3. Amino acid sequences ranging from 1 to 25 were signal peptide regions and 135-157 were transmembrane domains. The immunofluorescen assays showed that NcGRA3 protein behaved as an integrated transmembrane protein in PVs. Overall, characterization of the newly discovered NcGRA3 protein will lay a foundation for its function research in the further. Fascioliasis is caused by Fasciola species transmitted by freshwater Lymnaeidae snails and infecting herbivorous mammals and humans worldwide. In southern Asia, fascioliasis is a problem in livestock from the Near East to Bangladesh, where recent human infection reports are worrying. In this region, Fasciola gigantica is transmitted by species of the Radix auricularia superspecies group. In the densely populated northern Indian state of Uttar Pradesh, livestock appears infected throughout. The economic importance of buffaloes highlights the need to control their very high infection rates. In the Gorakhpur area, a molecular method based on the two specific primer sets of genomic DNA was applied to fasciolids from buffaloes slaughtered in local abattoirs and cercariae from R. acuminata snails from freshwater collections. PCR products and sequences demonstrated that the cercariae belonged to F. gigantica and that R. acuminata acts as vector for its transmission to buffaloes. The 72.0% rate found in one trancreases of rain events and floods suggest the need for control and surveillance efforts in this endemic area.