Smz745t

The specimens were observed under a stereomicroscope (Nikon SMZ745T, Nikon Corporation, Tokyo, Japan) and shot with a Canon digital camera (Canon EOS 1500D, Canon Inc., Tokyo, Japan) fitted on. Microscope images of the samples were taken by a Canon EOS 700D digital camera fitted on the Nikon ECLIPSE Ni compound microscope (Nikon, Japan). Measurements were taken with the Tarosoft (R) Image Frame Work (v.0.9.7). More than 30 morphological characteristics such as teliospores, urediniospores, and paraphyses were measured for each specimen. Photo plates were arranged by using Adobe Photoshop CS6 v. 13 (Adobe Systems Software Ireland Ltd, San Jose, USA). The different spore stages of rust fungi are designated by the following Roman numerals: spermogonia/spermatia (0), aecia/aeciospores (I), uredinia/urediniospores (II), telia/teliospores (III), and basidia/basidiospore (IV). We applied the definitions of spore stage based on Cummins and Hiratsuka [7 ], and followed morphological types of spermogonia designated by Hiratsuka and Hiratsuka [23 ].

To carry out this study, the NIKON SMZ 745T stereomicroscope (Nikon Corporation, Tokyo, Japan) was used at a maximum magnification of 75× and at a working distance of 115 mm (Figure 1). The device was used to examine dental surfaces with NCCLs and to acquire their 2D images, stored as TIFF, JPG and PNG files. The acquisition of 2D microscopic images, their storage and subsequent processing were carried out using NIS-A-AMEAS software (version 2021), the specific software platform for this imaging system. The successive acquisition of 2D images and their recomposition in order to obtain a 3D image of the examined dental surfaces were carried out with the help of NIS-A-EDF software [34 ].
In order to position the teeth for the stereomicroscopic examination, each tooth was fixed in silicone of increased consistency (Zetaplus L Intro KIT, Zhermack, Badia Polesine, Italy), leaving only the dental surface with the NCCL exposed for the examination. The stereomicroscopic examination was performed by a specialist of Engineering and Management of Technological Systems and a specialist of Biophysics. The interpretation and analysis of the obtained images were carried out by 2 specialists in the field of dental wear.

Fertilized chick eggs were purchased by Granja Santa Isabel (Córdoba, Spain) and incubated for three days at 38 °C in a humidified incubator until they were windowed. Five days later, methylcellulose discs with several treatments were implanted on the CAM, and incubated for an additional 48 h. For methylcellulose disc preparation, compounds were resuspended in a 1.2% solution of methylcellulose, and 10 μL drops of this solution were allowed to dry on a Teflon-coated surface in a laminar flow hood. DMSO containing disc was used as a negative control and aeroplysinin-1 (5.9 nmol/methylcellulose disc) was used as a positive control [11 (link)]. After incubation, CAM was examined under a stereomicroscope Nikon SMZ 745 T (Nikon, Tokyo, Japan), and photographed with a camera Nikon DS-Ri2 (Nikon, Tokyo, Japan). Results were analyzed by two independent observers. Each condition was scored as positive if a significant reduction or rebound of vessels in the treated area was observed.

Mated dams from both groups were sacrificed using cervical dislocation and were subjected to ova/embryo isolation on embryonic day 2 of pregnancy (ED2, approx. 32 h after presumed ovulation [15 (link),20 (link)]), on embryonic day 3 of pregnancy (ED3, approx. 56 h after presumed ovulation [16 (link)]), and on day 4 of pregnancy (ED4, approx. 80 h after presumed ovulation [11 (link),15 (link),16 (link)]). Recovery was performed by flushing oviducts and uteri using a flushing-holding medium [23 ] containing 1% bovine serum albumin (BSA, Sigma-Aldrich, St. Louis, MO, USA). Immediately after isolation, the collected material was subjected to stereomicroscopic classification (Nikon SMZ 745T, Nikon, Tokyo, Japan), and the numbers of isolated ova/embryos were determined. Data gathering was performed between the years 2011 and 2020. During this period, ova/embryo isolation was performed eight times on ED2, three times on ED3, and five times on ED4.

The animals were euthanized for further investigation at 8 weeks postoperative. The knee samples were photographed using a Nikon SMZ745T (Nikon, Tokyo, Japan). Each section was stained with hematoxylin & eosin and Safranin-O staining. The histological score for joint surface repair of each section was evaluated as previously reported10 (link)38 (link). The scoring was performed independently by two blinded observers. Quantitative histomorphometry of Safranin O-stained sections was performed using Image J software (National Institutes of Health, Bethesda, MD, USA) as previously reported20 (link). The total region of the articular cartilage repair site was defined and measured as the total repair tissue volume. The percent Safranin O-stained repaired tissue in the total soft tissue volume (%Saf-O) was obtained. The depth of the cartilage in the repaired tissue was measured at 8 weeks postoperative using Image J software. All data are presented as the means ± standard error of the mean (SEM).

One F. pusio male specimen was field collected in March 2011 in Pajarito, a locality from Medellin, Antioquia, Colombia (06°17′10.7″N, 75°36′43.7″W) at 1929 m above sea level and identified using morphological keys [[21] (link), [22] (link), [23] (link)]. Specimen collection was done under collection permit 16455 issued by CORANTIOQUIA on May 18, 2011. The specimen was preserved in 95% ethanol and stored at −20 °C until DNA extraction. Then, the specimen was dried at room temperature and photographed using a digital camera OPTIKAM Pro 3 connected to a trinocular stereomicroscope (Nikon SMZ745T, Nikon Corp., Tokyo, Japan) as support for species identification. The abdomen was dissected for total genomic DNA using a GenElute™ Mammalian Genomic DNA Miniprep Kit (Sigma-Aldrich) following the manufacturer's protocol. The DNA was quantified using the Qubit dsDNA High Sensitivity assay (Thermo Fisher Scientific, Waltham, MA, USA) and stored at −20 °C until further processing. Species identification was confirmed by comparing COI sequences at the barcode of life data systems BOLD and GenBank databases [24 (link),25 (link)].