Use of Clinically Oriented Laboratory Manuals in a Prosection Gross Anatomy Laboratory during COVID-19.

Gross anatomy is a critical course for the development of a variety of skills such as anatomical knowledge and spatial, critical, and clinical reasoning. There have been few attempts to integrate clinical applications in gross anatomy, with the majority of these being in the lecture hall and not in the laboratory. Clinical cases and guided questions were added to a laboratory manual (Clinically Oriented Laboratory Manuals (COLMs)) in a first-year medical gross anatomy prosection course during COVID-19. The effectiveness of the COLMs was analyzed using in-laboratory assessments between treatment and control groups, as well as student perceptions. There was no significant difference between in-lab assessment scores between students with or without the COLMs in 2020 (t1304.735= 0.647, p ;= 0.518). Student perceptions demonstrated that 61.6% strongly agreed or agreed that the COLMs were a good way to learn anatomy and 32.0% desired more COLMs in the lab. Overall, COLMs did not increase student knowledge by the end of a session. Students thought the COLMs were a good tool to learn anatomy because they helped become more clinically aware; however, students desired better implementation of the COLMs. The addition of COLMs in the laboratory is a potential method to address the need for clinical applications within the gross anatomy laboratory. Supplementary Information: The online version contains supplementary material available at 10.1007/s40670-023-01970-1.

Prosection or Dissection: Which is Best for Teaching the Anatomy of the Hand and Foot?

Due to the current trend of decreasing contact hours and less emphasis being given to the basic science courses in the pre-clinical years of medical education, it is essential that new approaches to teaching gross anatomy are investigated to ensure medical students are being adequately exposed to anatomical content. This study retrospectively analyzed practical examination data from four medical gross anatomy classes (N = 569) to ascertain which pedagogical approach, student participation in the dissection process, or interaction with prosected specimens is best for teaching the anatomy of the hand and foot. Data analysis involved the use of propensity score matching, a nonparametric preprocessing statistical approach which ensures accurate representation of the true treatment effect by balancing cohorts prior to statistical analysis. Statistical analysis indicated that those students who were exposed to the anatomy of the hand through interactions with prosected specimens performed 5.6% better (P = 0.012) while for the foot, students who interacted with prosections performed 13.0% better (P < 0.001). Although limited, data from this study suggest that utilizing prosections of the hand and foot seems to be a more advantageous pedagogical approach for teaching these regions than requiring students to dissect them.

Visualization of stereoscopic anatomic models of the paranasal sinuses and cervical vertebrae from the surgical and procedural perspective.

Recent improvements in three-dimensional (3D) virtual modeling software allows anatomists to generate high-resolution, visually appealing, colored, anatomical 3D models from computed tomography (CT) images. In this study, high-resolution CT images of a cadaver were used to develop clinically relevant anatomic models including facial skull, nasal cavity, septum, turbinates, paranasal sinuses, optic nerve, pituitary gland, carotid artery, cervical vertebrae, atlanto-axial joint, cervical spinal cord, cervical nerve root, and vertebral artery that can be used to teach clinical trainees (students, residents, and fellows) approaches for trans-sphenoidal pituitary surgery and cervical spine injection procedure. Volume, surface rendering and a new rendering technique, semi-auto-combined, were applied in the study. These models enable visualization, manipulation, and interaction on a computer and can be presented in a stereoscopic 3D virtual environment, which makes users feel as if they are inside the model. Anat Sci Educ 10: 598-606. © 2017 American Association of Anatomists.

The hLAMP-1-positive particulate matrix involved in cardiac mesenchyme formation in the chick does not include BMP-2.

Early heart development involves the transformation of endocardial cells in the atrioventricular canal and outflow tract regions into mesenchymal cells, a process called endocardial mesenchymal transformation (EMT). This process is initiated by factors, termed the particulate matrix, that are secreted by the myocardium. The particulate matrix causes a subset of endocardial cells to hypertrophy, lose their cell-cell contacts, form migratory processes, transform into mesenchymal cells, and migrate into the underlying endocardial cushions. The particulate matrix can be extracted using EDTA and the EDTA extract can initiate the EMT process. Earlier reports from our laboratory have shown that the particulate matrix can be detected with the hLAMP-1 antibody in immunostaining and Western blot analysis. In addition, similar proteins have been isolated from the growth media of stage 15-16 chick embryo myocardial cultures (MyoCM). Since other investigators have identified a possible role for bone morphogenetic protein (BMP)-2 during the EMT process in the heart, we asked whether BMP-2 is a part of the chick hLAMP-1-positive particulate matrix. To answer this question, we double stained stage 15-16 chick embryo sections with hLAMP-1 and BMP-2 antibodies. We found that BMP-2 signals do not colocalize with hLAMP-1-stained particles. In addition, using immunoprecipitation-Western blot analysis, we demonstrated no association of BMP-2 with the hLAMP-1-bound fraction of the EDTA extract or MyoCM. Our results indicate that BMP-2 is not a component of the hLAMP-1-positive particulate matrix in the chick.

Frog virus 3 ORF 53R, a putative myristoylated membrane protein, is essential for virus replication in vitro.

Although previous work identified 12 complementation groups with possible roles in virus assembly, currently only one frog virus 3 protein, the major capsid protein (MCP), has been linked with virion formation. To identify other proteins required for assembly, we used an antisense morpholino oligonucleotide to target 53R, a putative myristoylated membrane protein, and showed that treatment resulted in marked reductions in 53R levels and a 60% drop in virus titers. Immunofluorescence assays confirmed knock down and showed that 53R was found primarily within viral assembly sites, whereas transmission electron microscopy detected fewer mature virions and, in some cells, dense granular bodies that may represent unencapsidated DNA-protein complexes. Treatment with a myristoylation inhibitor (2-hydroxymyristic acid) resulted in an 80% reduction in viral titers. Collectively, these data indicate that 53R is an essential viral protein that is required for replication in vitro and suggest it plays a critical role in virion formation.

Inhibition of iridovirus protein synthesis and virus replication by antisense morpholino oligonucleotides targeted to the major capsid protein, the 18 kDa immediate-early protein, and a viral homolog of RNA polymerase II.

Frog virus 3 (FV3) is a large DNA virus that encodes approximately 100 proteins. Although the general features of FV3 replication are known, the specific roles that most viral proteins play in the virus life cycle have not yet been elucidated. To address the question of viral gene function, antisense morpholino oligonucleotides (asMOs) were used to transiently knock-down expression of specific viral genes and thus infer their role in virus replication. We designed asMOs directed against the major capsid protein (MCP), an 18 kDa immediate-early protein (18K) that was thought to be a viral regulatory protein, and the viral homologue of the largest subunit of RNA polymerase II (vPol-IIalpha). All three asMOs successfully inhibited translation of the targeted protein, and two of the three asMOs resulted in marked phenotypic changes. Knock-down of the MCP resulted in a marked reduction in viral titer without a corresponding drop in the synthesis of other late viral proteins. Transmission electron microscopy (TEM) showed that in cells treated with the anti-MCP MO assembly sites were devoid of viral particles and contained numerous aberrant structures. In contrast, inhibition of 18K synthesis did not block virion formation, suggesting that the 18K protein was not essential for replication of FV3 in fathead minnow (FHM) cells. Finally, consistent with the view that late viral gene expression is catalyzed by a virus-encoded or virus-modified Pol-II-like protein, knock-down of vPol-IIalpha triggered a global decline in late gene expression and virus yields without affecting the synthesis of early viral genes. Collectively, these results demonstrate the utility of using asMOs to elucidate the function of FV3 proteins.

Rana catesbeiana virus Z (RCV-Z): a novel pathogenic ranavirus.

A virus, designated Rana catesbeiana virus Z (RCV-Z), was isolated from the visceral tissue of moribund tadpoles of the North American bullfrog Rana catesbeiana. SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) analysis of viral proteins and sequence analysis of the amino terminal end of the major capsid protein showed that RCV-Z was similar to frog virus 3 (FV3) and other ranaviruses isolated from anurans and fish. However, analysis of restriction fragment profiles following digestion of viral genomic DNA with XbaI and BamHI indicated that RCV-Z was markedly different from FV3. Moreover, in contrast to FV3, RCV-Z contained a full-length copy of the viral homolog of eukaryotic initiation factor 2 alpha (eIF-2alpha). Experimental infection of bullfrog tadpoles with FV3 and RCV-Z demonstrated that RCV-Z was much more pathogenic than FV3, and that prior infection with FV3 protected them from subsequent RCV-Z induced mortality. Collectively, these results suggest that RCV-Z may represent a novel species of ranavirus capable of infecting frogs and that possession of a viral eIF-2alpha homolog (vIF-2alpha) correlates with enhanced virulence.

Identification of cDNA clones that encode hLAMP-1, a component of the particulate matrix associated with cardiac mesenchyme formation.

The heart extracellular matrix protein hLAMP-1 (lectin-associated matrix protein in the heart) is a component of the particulate matrix that activates the AV endothelium prior to its transformation into mesenchyme within the atrioventricular canal and proximal outflow tract of the heart. The role of hLAMP-1 in this process has yet to be determined, in part because of the limited amount of material available for analysis. To overcome this liability, a monoclonal antibody to hLAMP-1 has been used to recognize proteins expressed by cDNA clones. The isolated cDNAs encode an mRNA consistent with previously published immunohistochemical results. Expression profiles of these clones by in situ hybridization revealed staining in areas of the heart that expressed hLAMP-1 by immunocytochemistry. Taken together, these results suggest that these clones, which represent an expressed sequence tag for hLAMP-1, should provide the basis for isolating a full-length cDNA of hLAMP-1, a prerequisite for determining the functional role of this protein in heart development.