Journal: BMC Pulmonary Medicine

Article Title: Paradoxical effects of cigarette smoke and COPD on SARS-CoV-2 infection and disease

doi: 10.1186/s12890-021-01639-8

Figure Lengend Snippet: Cigarette Smoke (CS) extract blocks in vitro SARS-CoV-2 replication in Calu-3 cells. To investigate the effects of cigarette smoke (CS) exposure on SARS-CoV-2 infection, we performed a 72 h in vitro infection of Calu-3 cells, a line permissive to SARS-CoV2 infection and replication. Cells were sham- or CSE-treated for 24 h. Supernatants (SN) and cytoplasmic lysates were obtained from a cell aliquot to measure ACE2 levels by ELISA. Then, cells were infected with SARS-CoV-2 (2 h viral infection in normal media, then remove inoculum). Every 24 h cells were fixed for IF staining of infection, and cell lysates were harvested for SDS-PAGE and WB of viral nucleocapsid (N) protein. dsRNA intermediates arise during the replication of viral RNA (vRNA), and IF staining with dsRNA-specific J2 monoclonal Ab is a good marker for SARS-CoV-2 replication. A Nikon Ti2 automated microscopy was used to quantitatively measure infection, as seen by dsRNA signal. Whereas replication of vRNA peaked at 48 h ( A, B ) in sham-treated cells, CSE-treatment abrogated infection to levels below the limit of detection. Similar results were seen with WB for viral N protein, showing peak viral protein synthesis at 72 h ( C ). In C , immunoblots show two bands used for densitometry and separated with a horizontal white line, one for the N protein, and one for GADPH. The two bands were cropped from original gels that are available in a Supplementary repository. ACE2 protein levels were undetectable in the SN, but were unchanged in CSE-treated versus sham cell lysates (not shown). In summary, CSE-pre-exposure increased ACE2 levels but potently abrogated SARS-CoV-2 replication in this in vitro model. The figure is representative of three independent experiments

Article Snippet: A Nikon Ti2 automated microscopy was used to quantitatively measure infection, as seen by dsRNA signal.

Techniques: In Vitro, Infection, Enzyme-linked Immunosorbent Assay, Staining, SDS Page, Marker, Microscopy, Western Blot

Journal: Acta Crystallographica. Section F, Structural Biology Communications

Article Title: Approaches to automated protein crystal harvesting

doi: 10.1107/S2053230X14000387

Figure Lengend Snippet: Examples of semi-automated protein crystal-harvesting systems

Article Snippet: Microtool N/A Station for Automated Microscopy and Imaging (SAMI) FMP Products Inc., Greenwich, Connecticut, USA http://bit.ly/12uWFO9 Commercial product.

Techniques: Membrane, Micromanipulation, Microscopy, Imaging

Journal: Acta Crystallographica. Section F, Structural Biology Communications

Article Title: Approaches to automated protein crystal harvesting

doi: 10.1107/S2053230X14000387

Figure Lengend Snippet: Examples of protein crystal-harvesting mechanisms. (a) Microgripper (‘chopsticks’; http://bit.ly/13lEQh7; Ohara, Ohba, Tanikawa, Hiraki, Wakatsuki, Mizukawa et al., 2004 ▶); (b) microcapillary (ACH; http://bit.ly/1b572WM; Weidong et al., 2004 ▶); (c) microtool (UMR; http://bit.ly/17pws4n; Viola et al., 2011 ▶); (d) ‘loopless’ (CMM; http://bit.ly/1a20g6v; Kitago et al., 2005 ▶); (e) Acoustic Droplet Ejection (ADE; Soares et al., 2011 ▶); (f) microgripper (REACH; Heidari Khajepour et al., 2013 ▶); (g) optical tweezers (FOT; Hikima et al., 2013 ▶); (h) ‘crystal catcher’ (Kitatani et al., 2008 ▶); (i) non-crystallographic application of force-sensing microgripper (Femtotools; http://bit.ly/17yY9YK; Beyeler et al., 2007 ▶); and (j–l) manipulation of a crystal using the RodBot system (Tung et al. (2013 ▶, 2014 ▶). In (j), the RodBot begins to rotate, generating a vortex that is used to capture the crystal, in (k) the RodBot transports the captured captured crystal towards the loop and in (l) the RodBot stops rotating and the crystal is released into the loop for harvesting.

Article Snippet: Microtool N/A Station for Automated Microscopy and Imaging (SAMI) FMP Products Inc., Greenwich, Connecticut, USA http://bit.ly/12uWFO9 Commercial product.

Techniques:

Journal: Acta Crystallographica. Section F, Structural Biology Communications

Article Title: Approaches to automated protein crystal harvesting

doi: 10.1107/S2053230X14000387

Figure Lengend Snippet: Harvesting mechanisms used in current semi-autonomous protein crystal-harvesting systems and their relative advantages and disadvantages Examples of each harvesting mechanism are shown in Fig. 4 . More + symbols indicate a higher score or cost.

Article Snippet: Microtool N/A Station for Automated Microscopy and Imaging (SAMI) FMP Products Inc., Greenwich, Connecticut, USA http://bit.ly/12uWFO9 Commercial product.

Techniques: High Throughput Screening Assay, In Situ