Protocol to isolate nuclei from Chlamydomonas reinhardtii for ATAC sequencing.

The isolation of sufficient amounts of intact nuclei is essential to obtain high-resolution maps of chromatin accessibility via assay for transposase-accessible chromatin using sequencing (ATAC-seq). Here, we present a protocol for tag-free isolation of nuclei from both cell walled and cell wall-deficient strains of the green model alga Chlamydomonas reinhardtii at a suitable quality for ATAC-seq. We describe steps for nuclei isolation, quantification, and downstream ATAC-seq. This protocol is optimized to shorten the time of isolation and quantification of nuclei.

Novel Approaches for the Treatment of Pulmonary Fibrosis with Emphasis on the Role of Galectin-3 Inhibitors as a Potential Therapeutic Approach.

Pulmonary fibrosis is a disease affecting the lungs and the respiratory system that carries along a high fatality rate with no specific therapeutic approaches, making it a disorder sometimes termed as incurable. There have been various researches elaborating on the potential treatment and formulation approaches. Therapeutically effective drugs, new molecules, potential drug targets and novel delivery approaches have been identified. Recent findings suggest galectin-3 as a potential target to alleviate the condition by inhibition of the lectin. Certain molecules of galectin-3 have been discovered as promising therapeutic agents. These drug molecules have been administered either orally or through inhalation, and as of now, there is no candidate in the market to pose as a treatment for pulmonary fibrosis. There is a wide window to research and find novel dosage forms for the drug molecules to be presented as an efficacious and tolerable drug therapy against pulmonary fibrosis.

Orchestral manoeuvres in the light: crosstalk needed for regulation of the Chlamydomonas carbon concentration mechanism.

The inducible carbon concentration mechanism (CCM) in Chlamydomonas reinhardtii has been well defined from a molecular and ultrastructural perspective. Inorganic carbon transport proteins, and strategically located carbonic anhydrases deliver CO2 within the chloroplast pyrenoid matrix where Rubisco is packaged. However, there is little understanding of the fundamental signalling and sensing processes leading to CCM induction. While external CO2 limitation has been believed to be the primary cue, the coupling between energetic supply and inorganic carbon demand through regulatory feedback from light harvesting and photorespiration signals could provide the original CCM trigger. Key questions regarding the integration of these processes are addressed in this review. We consider how the chloroplast functions as a crucible for photosynthesis, importing and integrating nuclear-encoded components from the cytoplasm, and sending retrograde signals to the nucleus to regulate CCM induction. We hypothesize that induction of the CCM is associated with retrograde signals associated with photorespiration and/or light stress. We have also examined the significance of common evolutionary pressures for origins of two co-regulated processes, namely the CCM and photorespiration, in addition to identifying genes of interest involved in transcription, protein folding, and regulatory processes which are needed to fully understand the processes leading to CCM induction.

Methods for Identification and Validation of G-Quadruplex Sequences in Legumes.

Among the different secondary structures that DNA can adopt, G-quadruplex is a noncanonical form that has recently started to garner attention about the possible layers of regulation they could introduce in cellular processes. Here, we outline how the presence of G-quadruplexes can be probed in legumes and other plant genomes. This chapter describes various in silico approaches that can be utilized to identify putative G-quadruplex forming sequences (GQSes) and validate their formation through in vitro experimental approaches.