Vernier caliper

The twitching motility of the P. aeruginosa cells was assayed using the procedure of Singh et al. (2002) (link). An overnight culture of P. aeruginosa was diluted in fresh AB medium to an OD at 595 nm of approximately 0.1 and supplemented with LAE. The diluted solution (2 μL) was stab-inoculated on the bottom of a 1% AB agar plate, which was incubated at room temperature for 2 days. The zone of twitching was measured with a Vernier caliper (Mitutoyo, Tsukuba, Japan).

The authors created a cylindrical telescopic instrument (length of 23 cm, distal diameter of 1.5 mm) capable of being introduced into the knee through the arthroscopic portals (Figure 2). The instrument had a thin circular filament (0.02 mm in diameter with a maximal distal length of 3 mm) capable of sliding towards the tibiofemoral space permitting the vertical measurement of the JSW at the end of the instrument (Figure 2). The resolution of the device was 0.1 mm and had a vertical range of measurement between 1.5 and 7.5 mm. The instrument was made of surgical stainless steel.
The instrument was compared to a standardized digital Vernier caliper with a resolution of 0.05 mm (Mitutoyo, Japan). The caliper was fixed in a tibial phantom, permitting to measure different JSWs (Figure 2). The measurements of the JSW varied from 0 to 6 mm, each 0.05 mm, with the testing repeated three times.

Leishmania major parasites were harvested from the footpad-draining lymph nodes of L. major-infected BALB/c mice (after 3–4 weeks of infection). After 6 days of culture growth in M199 medium supplemented with 20% FCS, 100 U/ml penicillin (GIBCO), 100 µg/ml streptomycin (GIBCO), 4 mM NaHCO₃ (Merck), 0.0005% hemin (Sigma-Aldrich), and 0.1 mM adenosine (Sigma-Aldrich), the parasites were harvested by centrifugation (3,000 rpm, 10 min, 20°C) and washed in DMEM (GIBCO) medium. The metacyclic promastigotes were purified using a polysucrose (Sigma-Aldrich, St. Louis, MO, USA) gradient (20 and 10% in DMEM) by centrifugation (2,500 rpm, 15 min, 20°C, no break). Thereafter, the purified metacyclic promastigotes were washed twice in DMEM medium before being used for infection. The mice were infected with 2 × 10⁶L. major IR75 parasites in the hind footpads. The swelling of footpads of naïve and infected mice was monitored two times weekly using a Vernier caliper (Mitutoyo). Simultaneously, photographs of the naïve and infected footpads (front and side) were taken. The mice were sacrificed at indicated times by the use of CO₂ gas. From day 0, DKO mice were treated daily for a week with 1 µg of recombinant murine IL-12p70 (PeproTech EC, Ltd., London, UK) via the intraperitoneal route.

We collected leaves of 270 individuals (8–10 mature leaves per individual) from 32 populations located throughout the distribution range of the species (Fig. 1). A field permit of scientific collection (Official number SGPA/DGVS/12770/16) was issued by the Secretaria de Medio Ambiente y Recursos Naturales, of Mexico. Leaf samples were pressed and dried for further measurements in the lab and for herbarium specimens. For each specimen, we measured nine foliar traits: 1. total length including lamina and petiole (TL); 2. lamina length (LL); 3. maximum width of the lamina (MW); 4. petiole length (PL); 5. distance from the base to the maximum width of the lamina (DW); 6. petiole diameter (PD); 7. angle of the lamina apex (ALA); 8. ratio between MW and LL (WLR); and 9. ratio between DW and LL (DWLR) (Fig. 2A). All variables (except ALA) were measured using a Mitutoyo^® Vernier caliper (0.05 mm resolution) and are recorded in mm. For ALA, we used a Jeppesen PJ-1 Rotating Azimuth Plotter.

A calibrated Vernier caliper (Mitutoyo, Kawasaki, Japan) was used to determine the diameter and thickness of tablets.