Modified B-QUIET scoring of kidney images acquired by veterinary students in live canines with versus without sequential practice on ultrasound phantoms.

Ultrasound phantoms are training tools that can help students learn basic ultrasound principles. The purpose of this prospective cohort design study was to determine whether preclinical veterinary students in a curriculum with more phantom training sessions acquire better-quality ultrasound images of kidneys in live canines compared with students in a curriculum without sequential phantom training sessions. In clinical skills labs, 132 second-year (2VM) and 130 third-year (3VM) veterinary students obtained sagittal and transverse images of the left kidney of healthy, student-owned dogs. Images were graded on proper identification/orientation, technique, and image anatomy using a modified Brightness Mode Quality Ultrasound Imaging Examination Technique, a modified standardized ultrasound interpretation scale. A two-sample t-test was used to compare 2VM and 3VM performance. 2VM students were inaugural members of a redesigned curriculum and had previously participated in eight clinical skills labs involving hands-on ultrasound practice using phantoms and live animals prior to this study. The 3VM students were the final members of the prior curriculum and had previously participated in a single ultrasound lab using phantoms and a single ultrasound lab using live animals. For Identification/Orientation categories, 2VM students acquired slightly but statistically significantly better transverse images (P = 0.04). There were no significant differences between identification/orientation tasks for sagittal images or for technique and image anatomy categories. The findings indicate that future studies assessing more sensitive evaluation tools and serial evaluation of students may be beneficial in monitoring student competency and assist in evaluating the role of phantoms in ultrasound training in the veterinary curriculum.

What Is Your Diagnosis?

In collaboration with the American College of Veterinary Radiology.

Phylogenetic and full-length genome mutation analysis of SARS-CoV-2 in Indonesia prior to COVID-19 vaccination program in 2021.

BACKGROUND: Indonesia has started the big project of COVID-19 vaccination program since 13 January 2021 by employing the first shot of vaccine to the President of Indonesia as the outbreak and rapid transmission of COVID-19 have endangered not only Indonesian but the global health and economy. This study aimed to investigate the full-length genome mutation analysis of 166 Indonesian SARS-CoV-2 isolates as of 12 January 2021. RESULTS: All data of the isolates were extracted from the Global Initiative on Sharing All Influenza Data (GISAID) EpiCoV database. CoVsurver platform was employed to investigate the full-length genome mutation analysis of all isolates. This study also focused on the phylogeny analysis in unlocking the mutation of S protein in Indonesian SARS-CoV-2 isolates. WIV04 isolate that was originated from Wuhan, China was used as the virus reference according to the CoVsurver default. The result showed that a full-length genome mutation analysis of 166 Indonesian SARS-CoV-2 isolates was successfully generated. Every single mutation in S protein was described and then visualized by utilizing BioRender platform. Furthermore, it also found that D614G mutation appeared in 103 Indonesian SARS-CoV-2 isolates. CONCLUSIONS: To sum up, this study helped to observe the spread of COVID-19 transmission. However, it also proposed that the epidemiological surveillance and genomics studies might be improved on COVID-19 pandemic in Indonesia. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s42269-021-00657-0.

Brachial artery thrombosis in a dog causing monoparesis mimicking nerve sheath tumor.

There are few differential diagnoses for non-orthopedic thoracic limb lameness in adult dogs aside from nerve tumors and disk-associated nerve compression; this report introduces another etiology. A 9-year-old male castrated mixed dog presented with an episodic history of nonweight-bearing thoracic limb lameness. Additional clinical signs included an atrophied thoracic limb with cool paw pads and painful axillary region. Magnetic resonance imaging, computed tomography, ultrasound, and exploratory surgery confirmed a chronic thrombus of the right brachial artery. No underlying cause for the thrombus was identified. The dog has been successfully managed on long-term rivaroxaban and clopidogrel. Follow-up ultrasound of the thrombus suggested early remodeling.

Responses to acute infection with SARS-CoV-2 in the lungs of rhesus macaques, baboons and marmosets.

Non-human primate models will expedite therapeutics and vaccines for coronavirus disease 2019 (COVID-19) to clinical trials. Here, we compare acute severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection in young and old rhesus macaques, baboons and old marmosets. Macaques had clinical signs of viral infection, mild to moderate pneumonitis and extra-pulmonary pathologies, and both age groups recovered in two weeks. Baboons had prolonged viral RNA shedding and substantially more lung inflammation compared with macaques. Inflammation in bronchoalveolar lavage was increased in old versus young baboons. Using techniques including computed tomography imaging, immunophenotyping, and alveolar/peripheral cytokine response and immunohistochemical analyses, we delineated cellular immune responses to SARS-CoV-2 infection in macaque and baboon lungs, including innate and adaptive immune cells and a prominent type-I interferon response. Macaques developed T-cell memory phenotypes/responses and bystander cytokine production. Old macaques had lower titres of SARS-CoV-2-specific IgG antibody levels compared with young macaques. Acute respiratory distress in macaques and baboons recapitulates the progression of COVID-19 in humans, making them suitable as models to test vaccines and therapies.

The Effects of Content Delivery Methods on Ultrasound Knobology and Image Quality Recognition Training in First-Year Veterinary Students.

As new technologies are incorporated into the practice of veterinary medicine, it is imperative we utilize the most effective and impactful content delivery methods. Ultrasound technology has become more affordable and compact for veterinary hospitals, leading to easier incorporation into practice. This study compares three methods of delivering ultrasound knobology content to first-year veterinary students at Texas A&M University College of Veterinary Medicine and Biomedical Sciences. In a prospective study, first-year veterinary students were randomly selected to receive one of three content delivery methods: self-directed active learning (SDL), in-person instructor demonstration, or online module instruction. Knowledge acquisition was assessed using a 10-question quiz for short-term understanding followed by a 10-question quiz after a 6-week period to assess long-term knowledge retention. Student demographics were analyzed using the Chi-square test. Quiz scores were analyzed between groups using Kruskal-Wallis tests followed by Dunn's post-tests for multiple comparisons. Values of p ≤ .05 were considered significant. On the short-term and retention quiz questions, students participating in SDL scored significantly higher (10 [5-10]) than those receiving in-person instructor demonstration (9 [3-10] p = .01 and 8 [2-10] p = .0004, respectively) or the online module instruction training (Group C) (6 [1-10] p < 0.0001 and 8 [4-10] p < .001, respectively). Based on quiz scores, veterinary students exhibited better ultrasound knobology and image quality recognition proficiency immediately and at 6-weeks following SDL when compared with other content delivery methods. Self-directed learning methods are recommended when teaching ultrasonography to veterinary students.

SARS-CoV-2 infection leads to acute infection with dynamic cellular and inflammatory flux in the lung that varies across nonhuman primate species

There are no known cures or vaccines for COVID-19, the defining pandemic of this era. Animal models are essential to fast track new interventions and nonhuman primate (NHP) models of other infectious diseases have proven extremely valuable. Here we compare SARS-CoV-2 infection in three species of experimentally infected NHPs (rhesus macaques, baboons, and marmosets). During the first 3 days, macaques developed clinical signatures of viral infection and systemic inflammation, coupled with early evidence of viral replication and mild-to-moderate interstitial and alveolar pneumonitis, as well as extra-pulmonary pathologies. Cone-beam CT scans showed evidence of moderate pneumonia, which progressed over 3 days. Longitudinal studies showed that while both young and old macaques developed early signs of COVID-19, both groups recovered within a two-week period. Recovery was characterized by low-levels of viral persistence in the lung, suggesting mechanisms by which individuals with compromised immune systems may be susceptible to prolonged and progressive COVID-19. The lung compartment contained a complex early inflammatory milieu with an influx of innate and adaptive immune cells, particularly interstitial macrophages, neutrophils and plasmacytoid dendritic cells, and a prominent Type I-interferon response. While macaques developed moderate disease, baboons exhibited prolonged shedding of virus and extensive pathology following infection; and marmosets demonstrated a milder form of infection. These results showcase in critical detail, the robust early cellular immune responses to SARS-CoV-2 infection, which are not sterilizing and likely impact development of antibody responses. Thus, various NHP genera recapitulate heterogeneous progression of COVID-19. Rhesus macaques and baboons develop different, quantifiable disease attributes making them immediately available essential models to test new vaccines and therapies.

Reliability of interpretation of neurologic examination findings for the localization of vestibular dysfunction in dogs.

OBJECTIVE To estimate reliability of interpretation of neurologic examination findings for localization of vestibular dysfunction in dogs. DESIGN Cross-sectional study. ANIMALS 496 dogs that underwent MRI of the head for diagnosis of a neurologic problem between September 2011 and September 2015. PROCEDURES Medical records were reviewed and data collected regarding signalment and neurologic examination, MRI, and CSF findings. Independent observers interpreted the findings, and agreement was assessed for a subset of dogs. Distributions of variables were compared between dogs with and without a neurologic findings-based interpretation of vestibular disease. RESULTS 37% (185/496) of dogs had signs of vestibular dysfunction, of which 82% (151/185) had MRI abnormalities. In 73% (110/151) of dogs with MRI abnormalities, lesions involved central vestibular structures, and in 19% (29/151), lesions involved peripheral vestibular structures. On the basis of neurologic findings interpretation, 86% (160/185) of dogs were classified as having central vestibular dysfunction, and 61% (98/160) of these had an MRI-identified central vestibular lesion. Agreement among 3 independent observers was good (κ = 0.72) regarding use of neurologic examination findings to diagnose central versus peripheral vestibular dysfunction and very good (κ = 0.85) regarding use of MRI to diagnose peripheral vestibular lesions. Despite this agreement, only 29% (7/24) of dogs with a consensus clinical interpretation of peripheral vestibular dysfunction had MRI-identified peripheral lesions. CONCLUSIONS AND CLINICAL RELEVANCE Although interobserver agreement was good for distinguishing central from peripheral vestibular dysfunction in dogs through interpretation of neurologic examination findings, this interpretation did not agree with the MRI-based diagnosis.

Identification of the cytolinker plectin as a major early in vivo substrate for caspase 8 during CD95- and tumor necrosis factor receptor-mediated apoptosis.

Caspase 8 plays an essential role in the execution of death receptor-mediated apoptosis. To determine the localization of endogenous caspase 8, we used a panel of subunit-specific anti-caspase 8 monoclonal antibodies in confocal immunofluorescence microscopy. In the human breast carcinoma cell line MCF7, caspase 8 predominantly colocalized with and bound to mitochondria. After induction of apoptosis through CD95 or tumor necrosis factor receptor I, active caspase 8 translocated to plectin, a major cross-linking protein of the three main cytoplasmic filament systems, whereas the caspase 8 prodomain remained bound to mitochondria. Plectin was quantitatively cleaved by caspase 8 at Asp 2395 in the center of the molecule in all cells tested. Cleavage of plectin clearly preceded that of other caspase substrates such as poly(ADP-ribose) polymerase, gelsolin, cytokeratins, or lamin B. In primary fibroblasts from plectin-deficient mice, apoptosis-induced reorganization of the actin cytoskeleton, as seen in wild-type cells, was severely impaired, suggesting that during apoptosis, plectin is required for the reorganization of the microfilament system.

Not just scaffolding: plectin regulates actin dynamics in cultured cells.

Plectin, a major linker and scaffolding protein of the cytoskeleton, has been shown to be essential for the mechanical integrity of skin, skeletal muscle, and heart. Studying fibroblast and astroglial cell cultures derived from plectin (-/-) mice, we found that their actin cytoskeleton, including focal adhesion contacts, was developed more extensively than in wild-type cells. Also it failed to show characteristic short-term rearrangments in response to extracellular stimuli activating the Rho/Rac/Cdc42 signaling cascades. As a consequence, cell motility, adherence, and shear stress resistance were altered, and morphogenic processes were delayed. Furthermore, we show that plectin interacts with G-actin in vitro in a phosphatidylinositol-4,5-biphosphate-dependent manner and associates with actin stress fibers in living cells. The actin stress fiber phenotype of plectin-deficient fibroblasts could be reversed to a large degree by transient transfection of full-length plectin or plectin fragments containing the amino-terminal actin-binding domain (ABD). These results reveal a novel role of plectin as regulator of cellular processes involving actin filament dynamics that goes beyond its proposed role in scaffolding and mechanical stabilization of cells.

Targeted inactivation of plectin reveals essential function in maintaining the integrity of skin, muscle, and heart cytoarchitecture.

Previous studies suggest that plectin, a versatile cytoskeletal linker protein, has an important role in maintaining the structural integrity of diverse cells and tissues. To establish plectin's function in a living organism, we have disrupted its gene in mice. Plectin (-/-) mice died 2-3 days after birth exhibiting skin blistering caused by degeneration of keratinocytes. Ultrastructurally, hemidesmosomes and desmosomes appeared unaffected. In plectin-deficient mice, however, hemidesmosomes were found to be significantly reduced in number and apparently their mechanical stability was altered. The skin phenotype of these mice was similar to that of patients suffering from epidermolysis bullosa simplex (EBS)-MD, a hereditary skin blistering disease with muscular dystrophy, caused by defects in the plectin gene. In addition, plectin (-/-) mice revealed abnormalities reminiscent of minicore myopathies in skeletal muscle and disintegration of intercalated discs in heart. Our results clearly demonstrate a general role of plectin in the reinforcement of mechanically stressed cells. Plectin (-/-) mice will provide a useful tool for the study of EBS-MD, and possibly other types of plectin-related myopathies involving skeletal and cardiac muscle, in an organism amenable to genetic manipulation.

Expression of APP in transgenic mice: a comparison of neuron-specific promoters.

The beta-amyloid precursor protein (APP) carries mutations in codons 717 or 670/671, which cosegregate with familial forms of Alzheimer's disease (AD). As an initial step to study the related pathogenetic mechanisms in vivo we have generated transgenic mice expressing APP with these mutations. Several neuron-specific promoters were used to drive expression of human APP cDNAs. Only the Thy-1 promoter yielded transgene expression levels comparable to or above the endogenous mouse levels. Deletion of a 121 bp sequence from the 3' untranslated region of APP appeared to increase mRNA levels. Transgene mRNA was found throughout the brain with highest levels in hippocampus and cerebral cortex. Accordingly, human APP was detected in these regions by Western blotting. Protein levels paralleled mRNA levels reaching or exceeding the amount of endogenous APP. Variable reactivity of human APP in cell bodies was shown by immunocytochemistry. Although our initial histological examinations did not reveal any alterations characteristic of AD, further studied will be required.

[Follow-up of mastoiditis and mastoidectomy]. / Spätergebnisse nach Mastoiditis und Mastoidektomie.

BACKGROUND: To analyse the late results of mastoiditis and operative intervention as well as critical classification of our own indication for operation we invited 298 operated patients to follow-up examinations. PATIENTS AND METHODS: 218 of 298 patients operated on, took part in these examinations. The follow-up time ranged between 2 and 18 years. RESULTS: Altogether 311 ears were operated. Out of these, 69 patients had presented with classical mastoiditis and 242 with "masked" type. Microscopic examination of the operated ears showed that 284 (91.3%) had normal tympanic membranes, 13 (4.2%) had central tympanic membrane perforations with mucosal inflammation and 14 (4.5%) had residual retraction pockets or attic cholesteatomas. Hearing tests showed that 242 (77.8%) were normal, 56 (18%) had conductive deafness (with 32 of these ears having normal tympanic membranes), 10 (3.2%) had sensorineural deafness and 3 (1%) combined deafness. Schüller's radiographic views demonstrated compact mastoid processes in 48.7% of the reexamined ears and re-pneumatisation in 53%. CONCLUSION: The presented data of our re-examinations support our principle that prompt surgical treatment of all forms of mastoiditis is imperative.