Animal experimentation in oncology and radiobiology: Arguments for and against following a critical literature review

Despite the international 3Rs principles that recommends replacing, reducing and refining the use of animals in medical experimentation, it remains difficult to obtain funding in Canada for medical research that respects these principles, particularly with regard to replacement. This observation led our team to review the literature on the arguments for and against animal experimentation in the fields of oncology and radiobiology. This article presents a synthesis of these arguments. Using the method created by McCullough and colleagues to conduct critical reviews of the ethics literature, we analysed 25 texts discussing the arguments for and against animal experimentation in oncology and radiobiology. Six broad categories of arguments for animal experimentation and eleven categories of arguments against it emerged from our analyses. Furthermore, the arguments against animal testing are more convincing from both an empirical and normative perspective. Also, most arguments obtained are transferable in other fields of medicine. In addition to the literature review, a critical reflection was conducted and other arguments were discussed. It seems that a conservative culture persists in medical research, despite the scientific evidence and ethical arguments to the contrary. Malgré les principes internationaux des 3R qui recommande de remplacer, réduire et raffiner l’utilisation des animaux dans l’expérimentation médicale, il reste difficile d’obtenir au Canada le financement de recherches médicales qui respectent ces principes, notamment en ce qui concerne le remplacement. Ce constat a amené notre équipe à faire une revue de la littérature sur les arguments pour et contre l’expérimentation animale dans les domaines de l’oncologie et de la radiobiologie. Cet article présente une synthèse de ces arguments. En utilisant la méthode créée par McCullough et collègues pour réaliser des revues critiques de la littérature en éthique, nous avons analysé 25 écrits discutant des arguments pour et contre l’expérimentation animale en oncologie et en radiobiologie. Six grandes catégories d’arguments en faveur de l’expérimentation animale et onze catégories d’arguments contre celle-ci sont ressorties de nos analyses. En outre, les arguments contre l’expérimentation animale sont plus convaincants d’un point de vue empirique et normatif. De plus, la plupart des arguments obtenus sont transférables dans d’autres domaines de la médecine. En plus de l’analyse documentaire, une réflexion critique a été menée et d’autres arguments ont été discutés. Il semble qu’une culture conservatrice persiste dans la recherche médicale, malgré les preuves scientifiques et les arguments éthiques contraires

Sonoelastography of the Shoulder: A narrative review

Sonoelastography is a relatively new non-invasive imaging tool to assess the in vivo qualitative and quantitative biomechanical properties of various tissues. Two types of sonoelastography (SE) are commonly explored: strain and shear wave. Sonoelastography can be used in multiple medical subspecialties to assess pathological tissular changes by obtaining mechanical properties, shear wave speed, and strain ratio data. Although there are various radiological imaging methods, such as MRI or CT scan, to assess musculoskeletal structures (muscles, tendons, joint capsules), SE is more accessible since this approach is of low cost and does not involve radiation. As of 2018, SE has garnered promising data in multiple studies. Preliminary clinico-radiological correlations have been established to bridge tissue biomechanical findings with their respective clinical pathologies. Specifically, concerning the shoulder complex, recent findings have described mechanical tissue changes in shoulder capsulitis. The long head of the biceps and supraspinatus SE were among the recently studied structures with conditions regarding impingement, tendinosis, and tears. Since ultrasonography has established itself as an important tool in shoulder evaluation, it completes the history and physical examination skills of the clinicians. This study will provide an update on the most recent findings on SE of shoulder structures

Normal range of motion at the hip show different pressure behavior in the lateral and acetabular compartments: a cadaveric investigation

The techniques used previously to assess intracapsular pressures did not allow the assessment of pressure variations in both compartments throughout the entire range of motion without puncturing the capsular tissue. Our hypothesis was that the intra-capsular pressure would be different in the lateral and acetabular compartment depending on the movement assessed

Musculoskeletal ultrasound for 3D bone modeling: A preliminary study applied to lumbar vertebra

BACKGROUND: There is no non-invasive in vivo method to assess intervertebral kinematics. Current kinematics models are based on in vitro bone reconstructions from computed tomography (CT)-scan imaging, fluoroscopy and MRIs, which are either expensive or deleterious for human tissues. Musculoskeletal ultrasound is an accessible, easy to use and cost-effective device that allows high-resolution, real-time imaging of bone structure. OBJECTIVE: The aim of this preliminary study was to compare the concordance of 3D bone modeling of lumbar vertebrae between CT-scan and ultrasound imaging and to study the intra and inter-reliability of distances measured on 3D ultrasound bone models. METHODS: CT-scan, ultrasound, and in situ data of five lumbar vertebrae from the same human specimen were used. All vertebrae were scanned by tomography and a new musculoskeletal ultrasound procedure. Then, 3D bone modeling was created from both CT-scan and ultrasound image data set. Distances between anatomical bones landmarks were measured on the 3D models and compared to in situ measurements. © 2021-IOS Press. All rights reserved

The pharmacological regulation of cellular mitophagy

Small molecules are pharmacological tools of considerable value for dissecting complex biological processes and identifying potential therapeutic interventions. Recently, the cellular quality-control process of mitophagy has attracted considerable research interest; however, the limited availability of suitable chemical probes has restricted our understanding of the molecular mechanisms involved. Current approaches to initiate mitophagy include acute dissipation of the mitochondrial membrane potential (ΔΨm) by mitochondrial uncouplers (for example, FCCP/CCCP) and the use of antimycin A and oligomycin to impair respiration. Both approaches impair mitochondrial homeostasis and therefore limit the scope for dissection of subtle, bioenergy-related regulatory phenomena. Recently, novel mitophagy activators acting independently of the respiration collapse have been reported, offering new opportunities to understand the process and potential for therapeutic exploitation. We have summarized the current status of mitophagy modulators and analyzed the available chemical tools, commenting on their advantages, limitations and current applications

Klebsiella pneumoniae related community-acquired acute lower respiratory infections in CAMBODIA: clinical characteristics and treatment

<p>Abstract</p> <p>Background</p> <p>In many Asian countries, <it>Klebsiella pneumoniae </it>(KP) is the second pathogen responsible for community-acquired pneumonia. Yet, very little is known about <it>KP </it>etiology in ALRI in Cambodia, a country that has one of the weakest medical infrastructures in the region. We present here the first clinico-radiological description of <it>KP </it>community-acquired ALRI in hospitalized Cambodian patients.</p> <p>Methods</p> <p>Through ALRI surveillance in two provincial hospitals, <it>KP </it>was isolated from sputum and blood cultures, and identified by API20E gallery from patients ≥ 5 years-old with fever and respiratory symptoms onset ≤14 days. Antibiotics susceptibility testing was provided systematically to clinicians when bacteria were isolated. We collected patients' clinical, radiological and microbiological data and their outcome 3 months after discharge. We also compared <it>KP</it>-related with other bacteria-related ALRI to determine risk factors for <it>KP </it>infection.</p> <p>Results</p> <p>From April 2007 to December 2009, 2315 ALRI patients ≥ 5 years-old were enrolled including 587 whose bacterial etiology could be assigned. Of these, 47 (8.0%) had <it>KP </it>infection; their median age was 55 years and 68.1% were females. Reported prior medication was high (42.5%). Patients' chest radiographs showed pneumonia (61.3% including 39% that were necrotizing), preexisting parenchyma lesions (29.5%) and pleural effusions alone (4.5%) and normal parenchyma (4.5%). Five patients had severe conditions on admission and one patient died during hospitalization. Of the 39 patients that were hospital discharged, 14 died including 12 within 1 month after discharge. Only 13 patients (28%) received an appropriate antibiotherapy. Extended-spectrum beta-lactamases (ESBL) - producing strains were found in 8 (17.0%) patients. Female gender (Odds ratio (OR) 2.1; <it>p </it>= 0.04) and diabetes mellitus (OR 3.1; <it>p </it>= 0.03) were independent risk factors for <it>KP</it>-related ALRI.</p> <p>Conclusions</p> <p><it>KP </it>ALRI in Cambodia has high fatality rate, are more frequently found in women, and should be considered in diabetic patients. The extremely high frequency of ESBL-producing strains in the study is alarming in the context of uncontrolled antibiotic consumption and in absence of microbiology capacity in most public-sector hospitals.</p

Phosphorylation and Activation of the Plasma Membrane Na+/H+ Exchanger (NHE1) during Osmotic Cell Shrinkage

The Na+/H+ Exchanger isoform 1 (NHE1) is a highly versatile, broadly distributed and precisely controlled transport protein that mediates volume and pH regulation in most cell types. NHE1 phosphorylation contributes to Na+/H+ exchange activity in response to phorbol esters, growth factors or protein phosphatase inhibitors, but has not been observed during activation by osmotic cell shrinkage (OCS). We examined the role of NHE1 phosphorylation during activation by OCS, using an ideal model system, the Amphiuma tridactylum red blood cell (atRBC). Na+/H+ exchange in atRBCs is mediated by an NHE1 homolog (atNHE1) that is 79% identical to human NHE1 at the amino acid level. NHE1 activity in atRBCs is exceptionally robust in that transport activity can increase more than 2 orders of magnitude from rest to full activation. Michaelis-Menten transport kinetics indicates that either OCS or treatment with the phosphatase inhibitor calyculin-A (CLA) increase Na+ transport capacity without affecting transport affinity (Km = 44 mM) in atRBCs. CLA and OCS act non-additively to activate atNHE1, indicating convergent, phosphorylation-dependent signaling in atNHE1 activation. In situ 32P labeling and immunoprecipitation demonstrates that the net phosphorylation of atNHE1 is increased 4-fold during OCS coinciding with a more than 2-order increase in Na+ transport activity. This is the first reported evidence of increased NHE1 phosphorylation during OCS in any vertebrate cell type. Finally, liquid chromatography and mass spectrometry (LC-MS/MS) analysis of atNHE1 immunoprecipitated from atRBC membranes reveals 9 phosphorylated serine/threonine residues, suggesting that activation of atNHE1 involves multiple phosphorylation and/or dephosphorylation events

Cell Type-Specific Neuroprotective Activity of Untranslocated Prion Protein

Background: A key pathogenic role in prion diseases was proposed for a cytosolic form of the prion protein (PrP). However, it is not clear how cytosolic PrP localization influences neuronal viability, with either cytotoxic or anti-apoptotic effects reported in different studies. The cellular mechanism by which PrP is delivered to the cytosol of neurons is also debated, and either retrograde transport from the endoplasmic reticulum or inefficient translocation during biosynthesis has been proposed. We investigated cytosolic PrP biogenesis and effect on cell viability in primary neuronal cultures from different mouse brain regions. Principal Findings: Mild proteasome inhibition induced accumulation of an untranslocated form of cytosolic PrP in cortical and hippocampal cells, but not in cerebellar granules. A cyclopeptolide that interferes with the correct insertion of the PrP signal sequence into the translocon increased the amount of untranslocated PrP in cortical and hippocampal cells, and induced its synthesis in cerebellar neurons. Untranslocated PrP boosted the resistance of cortical and hippocampal neurons to apoptotic insults but had no effect on cerebellar cells. Significance: These results indicate cell type-dependent differences in the efficiency of PrP translocation, and argue that cytosolic PrP targeting might serve a physiological neuroprotective function

Pathogen-Mediated Proteolysis of the Cell Death Regulator RIPK1 and the Host Defense Modulator RIPK2 in Human Aortic Endothelial Cells

Porphyromonas gingivalis is the primary etiologic agent of periodontal disease that is associated with other human chronic inflammatory diseases, including atherosclerosis. The ability of P. gingivalis to invade and persist within human aortic endothelial cells (HAEC) has been postulated to contribute to a low to moderate chronic state of inflammation, although how this is specifically achieved has not been well defined. In this study, we demonstrate that P. gingivalis infection of HAEC resulted in the rapid cleavage of receptor interacting protein 1 (RIPK1), a mediator of tumor necrosis factor (TNF) receptor-1 (TNF-R1)-induced cell activation or death, and RIPK2, a key mediator of both innate immune signaling and adaptive immunity. The cleavage of RIPK1 or RIPK2 was not observed in cells treated with apoptotic stimuli, or cells stimulated with agonists to TNF-R1, nucleotide oligomerization domain receptor 1(NOD1), NOD2, Toll-like receptor 2 (TLR2) or TLR4. P. gingivalis-induced cleavage of RIPK1 and RIPK2 was inhibited in the presence of a lysine-specific gingipain (Kgp) inhibitor. RIPK1 and RIPK2 cleavage was not observed in HAEC treated with an isogenic mutant deficient in the lysine-specific gingipain, confirming a role for Kgp in the cleavage of RIPK1 and RIPK2. Similar proteolysis of poly (ADP-ribose) polymerase (PARP) was observed. We also demonstrated direct proteolysis of RIPK2 by P. gingivalis in a cell-free system which was abrogated in the presence of a Kgp-specific protease inhibitor. Our studies thus reveal an important role for pathogen-mediated modification of cellular kinases as a potential strategy for bacterial persistence within target host cells, which is associated with low-grade chronic inflammation, a hallmark of pathogen-mediated chronic inflammatory disorders