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BACKGROUND: The COVID-19 (C19) pandemic shocked primary care systems around the world. Those systems responded by supporting patients in the community, and acute care facilities in crisis. In Canada, the Patient Medical Home (PMH) is a widely adopted care model that aims to operationalize the tenets and principles of Primary Health Care (PHC) as developed since the Alma-Ata Declaration. This paper describes how personnel working in and with Primary Care Networks (PCNs) in Alberta, Canada deployed the PMH model and its underlying PHC principles to frame and respond to the C19 shock. METHODS: Using purposive and snowball sampling techniques, we interviewed 57 participants who worked in public health and primary care, including community-based family physicians. We used interpretive description to analyze the interviews. RESULTS: PCN staff and physicians described how the PMH model was foundational to normal operations, and how C19 responses were framed by the patient-centric, team-delivered, and continuous care principles the model shares with PHC. Specifically, participants described ensuring access to care, addressing the social determinants of health, being patient centered, and redeploying and expanding PHC teams to accomplish these goals. DISCUSSION: Delivering PHC through the PMH allowed physicians and allied health staff to deliver patient-centered, team-based, holistic bio-medical services to Albertans. In tailoring services to meet the specific social and health needs of the populations served by each PCN, healthcare providers were able to ensure relevant support remained available and accessible.
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Pandemias , Atención Primaria de Salud , Humanos , Alberta , Canadá , Atención Dirigida al Paciente , Médicos de FamiliaRESUMEN
OBJECTIVES: Differentiation between high-grade glioma (HGG) and post-treatment-related effects (PTRE) is challenging, but advanced imaging techniques were shown to provide benefit. We aim to investigate microstructure characteristics of metabolic compartments identified from amino acid PET and to evaluate the diagnostic potential of this multimodal and integrative O-(2-18F-fluoroethyl)-L-tyrosine-(FET)-PET and fast diffusion kurtosis imaging (DKI) approach for the detection of recurrence and IDH genotyping. METHODS: Fifty-nine participants with neuropathologically confirmed recurrent HGG (n = 39) or PTRE (n = 20) were investigated using static 18F-FET PET and a fast-DKI variant. PET and advanced diffusion metrics of metabolically defined (80-100% and 60-75% areas of 18F-FET uptake) compartments were assessed. Comparative analysis was performed using Mann-Whitney U tests with Holm-Sídák multiple-comparison test and Wilcoxon signed-rank test. Receiver operating characteristic (ROC) curves, regression, and Spearman's correlation analysis were used for statistical evaluations. RESULTS: Compared to PTRE, recurrent HGG presented increased 18F-FET uptake and diffusivity (MD60), but lower (relative) mean kurtosis tensor (rMKT60) and fractional anisotropy (FA60) (respectively p < .05). Diffusion metrics determined from the metabolic periphery showed improved diagnostic performance - most pronounced for FA60 (AUC = 0.86, p < .001), which presented similar benefit to 18F-FET PET (AUC = 0.86, p < .001) and was negatively correlated with amino acid uptake (rs = - 0.46, p < .001). When PET and DKI metrics were evaluated in a multimodal biparametric approach, TBRmax + FA60 showed highest diagnostic accuracy (AUC = 0.93, p < .001), which improved the detection of relapse compared to PET alone (difference in AUC = 0.069, p = .04). FA60 and MD60 distinguished the IDH genotype in the post-treatment setting. CONCLUSION: Detection of glioma recurrence benefits from a multimodal and integrative PET/DKI approach, which presented significant diagnostic advantage to the assessment based on PET alone. CLINICAL RELEVANCE STATEMENT: A multimodal and integrative 18F-FET PET/fast-DKI approach for the non-invasive microstructural characterization of metabolic compartments provided improved diagnostic capability for differentiation between recurrent glioma and post-treatment-related changes, suggesting a role for the diagnostic workup of patients in post-treatment settings. KEY POINTS: ⢠Multimodal PET/MRI with integrative analysis of 18F-FET PET and fast-DKI presents clinical benefit for the assessment of CNS cancer, particularly for the detection of recurrent high-grade glioma. ⢠Microstructure markers of the metabolic periphery yielded biologically pertinent estimates characterising the tumour microenvironment, and, thereby, presented improved diagnostic accuracy with similar accuracy to amino acid PET. ⢠Combined 18F-FET PET/fast-DKI achieved the best diagnostic performance for detection of high-grade glioma relapse with significant benefit to the assessment based on PET alone.
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Neoplasias Encefálicas , Glioma , Humanos , Glioma/diagnóstico por imagen , Glioma/patología , Imagen por Resonancia Magnética/métodos , Tomografía de Emisión de Positrones/métodos , Enfermedad Crónica , Tirosina , Recurrencia , Neoplasias Encefálicas/diagnóstico por imagen , Neoplasias Encefálicas/patología , Microambiente TumoralRESUMEN
AIM: To compare measurements on images obtained by magnetic resonance imaging (MRI) and cone beam CT (CBCT) for height, width, and area in alveolar bone sites in human jaw specimens. MATERIAL AND METHODS: Forty edentulous alveolar posterior sites in human cadaver specimens were imaged using CBCT scanners, and with zero-echo-time MRI (ZTE-MRI). Semi-automatic volume registration was performed to generate representative coronal sections of the sites related to implant planning. ZTE-MRI sections were also presented after grayscale inversion (INV MRI). Three observers measured bone height, bone width 5 mm from the alveolar crest, and bone area stretching from the width measurement to the top of the alveolar crest in the images. Interobserver agreement was assessed by intra-class correlation coefficients (ICC). The measurements were analyzed using two-way repeated measures ANOVA factoring observer and image type. RESULTS: ICC was >0.95 for bone height, width, and bone area. No significant differences among observers (p = 0.14) or image type (p = 0.60) were found for bone height. For bone width, observer (p = 0.14) was not a significant factor, while ZTE-MRI produced width estimates that were significantly different and systematically smaller than CBCT-based estimates (p ≤ 0.001). Observer (p = 0.06) was not a significant factor regarding the bone area measurements, contrary to the imaging type where ZTE-MRI led to significantly smaller area estimates than CBCT (p ≤ 0.001). CONCLUSION: Bone height measurements were essentially equivalent using CBCT and MRI. This was found regardless of grayscale choice for the MRI. However, ZTE-MRI resulted in smaller estimates of bone width and area.
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Implantes Dentales , Humanos , Proceso Alveolar/diagnóstico por imagen , Tomografía Computarizada de Haz Cónico/métodos , Cadáver , Imagen por Resonancia MagnéticaRESUMEN
Glioblastoma (GBM) is an aggressive brain tumor with a median survival of 15 months and has limited treatment options. Immunotherapy with checkpoint inhibitors has shown minimal efficacy in combating GBM, and large clinical trials have failed. New immunotherapy approaches and a deeper understanding of immune surveillance of GBM are needed to advance treatment options for this devastating disease. In this study, we used two preclinical models of GBM: orthotopically delivering either GBM stem cells or employing CRISPR-mediated tumorigenesis by adeno-associated virus, to establish immunologically proficient and non-inflamed tumors, respectively. After tumor development, the innate immune system was activated through long-term STING activation by a pharmacological agonist, which reduced tumor progression and prolonged survival. Recruitment and activation of cytotoxic T-cells were detected in the tumors, and T-cell specificity towards the cancer cells was observed. Interestingly, prolonged STING activation altered the tumor vasculature, inducing hypoxia and activation of VEGFR, as measured by a kinome array and VEGF expression. Combination treatment with anti-PD1 did not provide a synergistic effect, indicating that STING activation alone is sufficient to activate immune surveillance and hinder tumor development through vascular disruption. These results guide future studies to refine innate immune activation as a treatment approach for GBM, in combination with anti-VEGF to impede tumor progression and induce an immunological response against the tumor.
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Neoplasias Encefálicas , Glioblastoma , Humanos , Neoplasias Encefálicas/inmunología , Neoplasias Encefálicas/metabolismo , Glioblastoma/inmunología , Glioblastoma/metabolismo , Inmunoterapia/métodos , Microambiente Tumoral , Inmunidad InnataRESUMEN
Background: The osteocutaneous radial forearm (OCRF) flap is a variation of the traditional radial forearm flap with incorporation of an anterolateral segment of corticocancellous bone of the radius, periosteum, and overlying skin. The OCRF flap is indicated in traumatic injuries or extirpation defects with segmental bone loss and is well suited to foot and ankle reconstruction due to its thin pliable skin. Methods: In this single-center case series, a retrospective review was conducted to identify patients who underwent OCRF free flap for foot and ankle reconstruction that required harvest of more than 50% of the cross-sectional area of the radius with prophylactic volar locked plating of the donor site. Outcome measures included flap failure rates, postoperative fracture, thrombotic events, time to follow-up, and time to full weightbearing. Flap harvest technique is extensively discussed. Results: Six cases were included in this series. There were no flap failures or thrombotic events. Recipient site healing was confirmed in all patients, with partial distal skin paddle loss in one patient requiring operative debridement. No patients sustained donor site complications or functional impairment. Full lower extremity weightbearing was achieved at 12.4 ± 3.3 weeks after surgery. Conclusions: The OCRF free flap transfer provides a reliable means of obtaining thin, supple soft tissue coverage with a large, vascularized segment of bone for reconstruction in the foot and ankle. Here, we describe use of more than 50% of the cross-sectional area of the radius with volar locked prophylactic plating. These updates expand use of this reconstructive technique.
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Background: Radiocapitellar arthroplasty fills a treatment void for young patients who experience isolated capitellar fractures or radiocapitellar osteoarthritis who are not candidates for total elbow arthroplasty. The outcomes of this procedure are sparsely reported. We designed a meta-analysis to determine the utility of radiocapitellar arthroplasty with respect to functional and patient reported outcomes. Methods: The PubMed database was searched for relevant studies. Only studies published in English language that assessed patient reported outcomes following radiocapitellar arthroplasty were included in this study. Reporting followed the Preferred Reporting Items for Systematic Reviews and Meta-analyses for 2020. Results: The initial review yielded 562 studies that met the criteria. After excluding duplications and confounding factors, eight case series were identified for review. Of the eight studies, seven were eligible for inclusion in the meta-analysis for Mayo Elbow Performance Score, flexion-extension arc, and pronation-supination arc. The pooled standard mean difference was found to be statistically significant between pre-operative and post-operative outcomes for Mayo Elbow Performance Score (SMD = 3.04, 95% CI [2.40, 3.67]), flexion-extension arc (SMD = 1.28, 95% CI [0.73, 1.83]), and pronation-supination arc (SMD = 0.81, 95% CI [0.43, 1.18]). Cochran's Q-test and I2 statistics indicated statistically significant heterogeneity for Mayo Elbow Performance Score (p = .04, I2 = 54%) and flexion-extension arc (p < .01, I2 = 67%). Conclusions: Patients undergoing radiocapitellar arthroplasty showed statistically significant improvements in flexion-extension arc, pronation-supination arc, and Mayo Elbow Performance Scores compared to pre-operative measures.
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Recent studies have shown significant changes to brain microstructure during sleep and anesthesia. In vivo optical microscopy and magnetic resonance imaging (MRI) studies have attributed these changes to anesthesia and sleep-related modulation of the brain's extracellular space (ECS). Isoflurane anesthesia is widely used in preclinical diffusion MRI (dMRI) and it is therefore important to investigate if the brain's microstructure is affected by anesthesia to an extent detectable with dMRI. Here, we employ diffusion kurtosis imaging (DKI) to assess brain microstructure in the awake and anesthetized mouse brain (n = 22). We find both mean diffusivity (MD) and mean kurtosis (MK) to be significantly decreased in the anesthetized mouse brain compared with the awake state (p < 0.001 for both). This effect is observed in both gray matter and white matter. To further investigate the time course of these changes we introduce a method for time-resolved fast DKI. With this, we show the time course of the microstructural alterations in mice (n = 5) as they transition between states in an awake-anesthesia-awake paradigm. We find that the decrease in MD and MK occurs rapidly after delivery of gas isoflurane anesthesia and that values normalize only slowly when the animals return to the awake state. Finally, time-resolved fast DKI is employed in an experimental mouse model of brain edema (n = 4), where cell swelling causes the ECS volume to decrease. Our results show that isoflurane affects DKI parameters and metrics of brain microstructure and point to isoflurane causing a reduction in the ECS volume. The demonstrated DKI methods are suitable for in-bore perturbation studies, for example, for investigating microstructural modulations related to sleep/wake-dependent functions of the glymphatic system. Importantly, our study shows an effect of isoflurane anesthesia on rodent brain microstructure that has broad relevance to preclinical dMRI.
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The Locus Coeruleus (LC) is in the brainstem and supplies key brain structures with noradrenaline, including the forebrain and hippocampus. The LC impacts specific behaviors such as anxiety, fear, and motivation, as well as physiological phenomena that impact brain functions in general, including sleep, blood flow regulation, and capillary permeability. Nevertheless, the short- and long-term consequences of LC dysfunction remain unclear. The LC is among the brain structures first affected in patients suffering from neurodegenerative diseases such as Parkinson's disease and Alzheimer's Disease, hinting that LC dysfunction may play a central role in disease development and progression. Animal models with modified or disrupted LC function are essential to further our understanding of LC function in the normal brain, the consequences of LC dysfunction, and its putative roles in disease development. For this, well-characterized animal models of LC dysfunction are needed. Here, we establish the optimal dose of selective neurotoxin N-(2-chloroethyl)-N-ethyl-bromo-benzylamine (DSP-4) for LC ablation. Using histology and stereology, we compare LC volume and neuron number in LC ablated (LCA) mice and controls to assess the efficacy of LC ablation with different numbers of DSP-4 injections. All LCA groups show a consistent decrease in LC cell count and LC volume. We then proceed to characterize the behavior of LCA mice using a light-dark box test, Barnes maze test, and non-invasive sleep-wakefulness monitoring. Behaviorally, LCA mice differ subtly from control mice, with LCA mice generally being more curious and less anxious compared to controls consistent with known LC function and projections. We note an interesting contrast in that control mice have varying LC size and neuron count but consistent behavior whereas LCA mice (as expected) have consistently sized LC but erratic behavior. Our study provides a thorough characterization of an LC ablation model, firmly consolidating it as a valid model system for the study of LC dysfunction.
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The defensive activation theory (DAT) was recently proposed to explain the biological function of dreaming. Briefly, DAT states that dreams are primarily visual to prevent plastic take-over of an otherwise inactive visual cortex during sleep. Evidence to support the DAT revolve around the interplay between dream activity (REM%) and cortical plasticity found in evolutionary history, primate studies, and coinciding decline in human cortical plasticity and REM% with age. As the DAT may prove difficult to test experimentally, we investigate whether further support for the DAT can be found in the literature. Plasticity and REM sleep are closely linked to functions of the Locus Coeruleus (LC). We therefore review existing knowledge about the LC covering LC stability with age, and the role of the LC in the plasticity of the visual cortex. Recent studies show the LC to be more stable than previously believed and therefore, the LC likely supports the REM% and plasticity in the same manner throughout life. Based on this finding, we review the effect of aging on REM% and visual cortex plasticity. Here, we find that recent, weighty studies are not in complete agreement with the data originally provided as support for DAT. Results from these studies, however, are not in themselves irreconcilable with the DAT. Our findings therefore do not disprove the DAT. Importantly, we show that the LC is involved in all mechanisms central to the DAT. The LC may therefore provide an experimental window to further explore and test the DAT.
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PURPOSE: The effective integration of primary care into public health responses to the COVID-19 pandemic, particularly through data sharing, has received some attention in the literature. However, the specific policies and structures that facilitate this integration are understudied. This paper describes the experiences of clinicians and administrators in Alberta, Canada as they built a data bridge between primary care and public health to improve the province's community-based response to the pandemic. METHODS: Fifty-seven semistructured qualitative interviews were conducted with a range of primary care and public health stakeholders working inside the Calgary Health Zone. Interpretive description was used to analyze the interviews. RESULTS: SARS-CoV-2 test results produced by the local public laboratory were, initially, only available to central public health clinicians and not independent primary care physicians. This enabled centrally managed contact tracing but meant primary care physicians were unaware of their patients' COVID-19 status and unable to offer in-community follow-up care. Stakeholders from both central public health and independent primary care were able to leverage a policy commitment to the Patient Medical Home (PMH) care model, and a range of existing organizational structures, and governance arrangements to create a data bridge that would span the gap. CONCLUSIONS: Primary care systems looking to draw lessons from the data bridge's construction may consider ways to: leverage care model commitments to integration and adjust or create organization and governance structures which actively draw together primary care and non-primary care stakeholders to work on common projects. Such policies and structures develop trusting relationships, open the possibility for champions to emerge, and create the spaces in which integrative improvisation can take place.
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COVID-19 , Humanos , Salud Pública , Pandemias , SARS-CoV-2 , Política de SaludRESUMEN
BACKGROUND: Spurred by the Coronavirus infectious disease 2019 pandemic, aerosol containment devices (ACDs) were developed to capture infectious respiratory aerosols generated by patients at their source. Prior reviews indicated that such devices had low evidence of effectiveness, but did not address how ACDs should be evaluated, how well they should perform, nor have clearly defined performance standards. Towards developing design criteria for ACDs, two questions were posed: 1) What characteristics have guided the design of ACDs? 2) How have these characteristics been evaluated? METHODS: A scoping review was performed consistent with PRISMA guidelines. Data were extracted with respect to general study information, intended use of the device, device design characteristics and evaluation. RESULTS: Fifty-four articles were included. Evaluation was most commonly performed with respect to device aerosol containment (n = 31, 61%), with only 5 (9%), 3 (6%) and 8 (15%) formally assessing providing experience, patient experience and procedure impact, respectively. Nearly all of the studies that explored provider experience and procedure impact studied intubation. Few studies provided a priori performance criteria for any evaluation metric, or referenced any external guidelines by which to bench mark performance. CONCLUSION: With respect to aerosol containment, ACDs should reduce exposure among HCP with the device compared with the absence of the device, and provide ≥90% reduction in respirable aerosols, equivalent in performance to N95 filtering facepiece respirators, if the goal is to reduce reliance on personal protective equipment. The ACD should not increase awkward or uncomfortable postures, or adversely impact biomechanics of the procedure itself as this could have implications for procedure outcomes. A variety of standardized instruments exist to assess the experience of patients and healthcare personnel. Integration of ACDs into routine clinical practice requires rigorous studies of aerosol containment and the user experience.
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COVID-19 , Aerosoles y Gotitas Respiratorias , Humanos , COVID-19/epidemiología , COVID-19/prevención & control , Equipo de Protección Personal , Intubación Intratraqueal/métodos , Diseño de EquipoRESUMEN
OBJECTIVES: Dental materials, including orthodontic appliances and implants, are commonly mentioned as a possible source of artefacts in magnetic resonance imaging (MRI). The aim of the present study was to undertake a systematic review of the relevant literature on MR image artefacts due to dental materials, limited to orthodontic appliances and implant-supported dental prosthesis, on both technical and diagnostic levels. METHODS: The MEDLINE (PubMed) bibliographic database was searched up to September 2020. The search was limited to studies published in English, using the search string: (MRI or magnetic resonance) and (artefact or artifact) and (dental or ortho or implant or restoration or restorative). The studies were assessed independently by three reviewers, focusing on the following parameters: MRI sequences, tested materials, assessed parameters, efficacy level and outcome. RESULTS: The search strategy yielded 31 studies, which were included in this systematic review. These studies showed that metallic dental materials, commonly present in orthodontic appliances and implant-supported dental prosthesis led to diverse types/severities of artefacts in MR images. Fifteen studies were in vivo, based on human subjects. The studies differed substantially in terms of tested materials, assessed parameters, and outcome measurements. CONCLUSIONS: Metallic dental materials cause artefacts of diverse types and severities in MR images of the head and neck region. However, the diagnostic relevance of the investigated artefacts for the diverse MRI applications is yet to be studied.
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Artefactos , Aparatos Ortodóncicos , Humanos , Aparatos Ortodóncicos/efectos adversos , Imagen por Resonancia Magnética/efectos adversos , Imagen por Resonancia Magnética/métodos , Cabeza , Materiales DentalesRESUMEN
OBJECTIVES: To compare alveolar bone height and width measurements from zero-echo-time MRI (ZTE-MRI) and cone beam CT (CBCT), in human specimens. MATERIAL AND METHODS: Twenty posterior edentulous sites in human cadaver specimens were imaged with CBCT and ZTE-MRI. Bone height and width at 1, 3, 5, 7 and 9 mm from the top of the alveolar ridge was measured by two trained observers in cross-sections of a site where an implant was to be planned. Twenty percent of the sample was measured in duplicate to assess method error and intra-observer reproducibility (ICC). The differences between CBCT and ZTE-MRI measurements were compared (t-test). RESULTS: Inter- and intra-observer reproducibility was >0.90. The method error (average between observers) for bone height was 0.45 mm and 0.39 mm, and for bone width (average) was 0.52 mm and 0.80 mm (CBCT and ZTE-MRI, respectively). The majority of the bone measurement differences were statistically insignificant, except bone width measurements at 5 mm (p ≤ .05 for both observers). Mean measurement differences were not larger than the method error. CONCLUSION: ZTE-MRI is not significantly different from CBCT when comparing measurements of alveolar bone height and width.
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Proceso Alveolar , Tomografía Computarizada de Haz Cónico , Humanos , Reproducibilidad de los Resultados , Proceso Alveolar/diagnóstico por imagen , Proceso Alveolar/anatomía & histología , Tomografía Computarizada de Haz Cónico/métodos , Imagen por Resonancia Magnética , CadáverRESUMEN
BACKGROUND: The first wave of COVID-19 in Calgary, Alberta accelerated the integration of primary care with the province's centrally managed health system. This integration aimed to deliver wraparound in-community patient care through two interventions that combined to create the COVID-19 Integrated Pathway (CIP). The CIP's interventions were: 1) a data sharing platform that ensured COVID-19 test results were directly available to family physicians (FPs), and 2) a clinical algorithm that supported FPs in delivering in-community follow up to improve patient outcomes. We describe the CIP function and its capacity to facilitate FP follow-up with COVID-19 patients and evaluate its impact on Emergency Department (ED) visits and hospitalization. METHOD: We generated descriptive statistics by analyzing data from a Calgary Zone hub clinic called the Calgary COVID-19 Care Clinic (C4), provincially maintained records of hospitalization, ED visits, and physician claims. RESULTS: Between Apr. 16 and Sep. 27, 2020, 7289 patients were referred by the Calgary Public Health team to the C4 clinic. Of those, 48.6% were female, the median age was 37.4 y. 97% of patients had at least one visit with a healthcare professional, where follow-up was conducted using the CIP's algorithm. 5.1% of patients visited an ED and 1.9% were hospitalized within 30 days of diagnosis. 75% of patients had a median of 4 visits with their FP. DISCUSSION: Our data suggest that information exchange between Primary Care (PC) and central systems facilitates primary care-based management of patients with COVID-19 in the community and has potential to reduce acute care visits.
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COVID-19 , Médicos , Adulto , Femenino , Humanos , Masculino , COVID-19/terapia , Hospitalización , Atención Primaria de Salud , Cambio Social , Salud PúblicaRESUMEN
Medical imaging techniques are widely used in preclinical research as diagnostic tools to detect physiological abnormalities and assess the progression of neurovascular disease in animal models. Despite the wealth of imaging options in magnetic resonance imaging (MRI), interpretation of imaging-derived parameters regarding underlying tissue properties is difficult due to technical limitations or lack of parameter specificity. To address the challenge of interpretation, we present an animal preparation protocol to achieve quantitative measures from both MRI and advanced optical techniques, including laser speckle contrast imaging and two-photon microscopy, in murine models. In this manner, non-translatable methods support and improve interpretation of less specific, translatable methods, i.e., MRI. Combining modalities for improved clinical interpretation involves satisfying the requirements of various methods. Furthermore, physiology unperturbed by anesthetics is a prerequisite for the strategy to succeed. Awake animal imaging with restraint provides an alternative to anesthesia and facilitates translatability of cerebral measurements. The method outlines design requirements for the setup and a corresponding reproducible surgical procedure for implanting a 3D printed head holder and cranial window to enable repeated multimodal imaging. We document the development, application, and validation of the method and provide examples confirming the usefulness of the design in acquiring high quality data from multiple modalities for quantification of a wide range of metrics of cerebral physiology in the same animal. The method contributes to preclinical small animal imaging, enabling sequential imaging of previously mutually exclusive techniques.
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The naturally occurring ectoparasite salmon lice (Lepeophtherirus salmonis) poses a great challenge for the salmon farming industry, as well as for wild salmonids in the Northern hemisphere. To better control the infestation pressure and protect the production, there is a need to provide fish farmers with sensitive and efficient tools for rapid early detection and monitoring of the parasitic load. This can be achieved by targeting L. salmonis DNA in environmental samples. Here, we developed and tested a new L. salmonis specific DNA-based assay (qPCR assay) for detection and quantification from seawater samples using an analytical pipeline compatible with the Environmental Sample Processor (ESP) for autonomous water sample analysis of gene targets. Specificity of the L. salmonis qPCR assay was demonstrated through in-silico DNA analyses covering sequences of different L. salmonis isolates. Seawater was spiked with known numbers of nauplii and copepodite free-swimming (planktonic) stages of L. salmonis to investigate the relationship with the number of marker gene copies (MGC). Finally, field samples collected at different times of the year in the vicinity of a salmon production farm in Western Norway were analyzed for L. salmonis detection and quantification. The assay specificity was high and a high correlation between MGC and planktonic stages of L. salmonis was established in the laboratory conditions. In the field, L. salmonis DNA was consequently detected, but with MGC number below that expected for one copepodite or nauplii. We concluded that only L. salmonis tissue or eDNA residues were detected. This novel study opens for a fully automatized L. salmonis DNA quantification using ESP robotic to monitor the parasitic load, but challenges remain to exactly transfer information about eDNA quantities to decisions by the farmers and possible interventions.
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Copépodos , ADN Ambiental , Enfermedades de los Peces , Animales , Acuicultura , Copépodos/genética , ADN Ambiental/genética , Enfermedades de los Peces/diagnóstico , Enfermedades de los Peces/parasitología , Salmón/parasitología , AguaRESUMEN
Traditionally, preclinical magnetic resonance imaging (MRI) has been performed in anesthetized animals. However, anesthesia has been shown to perturb normal brain function and physiology. Such effects limit our ability to detect subtle physiological alterations in disease models and treatment studies, thus hampering discovery and compromising generality of findings. Therefore, methods for awake animal MRI are needed to study the rodent brain in its natural physiological state, free of anesthetics. Current setups for awake animal MRI rely on restraining systems to avoid animal movement during scanning. To reduce restraint stress, animals are habituated to the scanner environment prior to MRI data collection. To date, however, most awake MRI studies employ male rodents only. This is a fundamental limitation as results obtained may be pertinent only to half of the population. We characterized training and habituation responses of male and female mice to provide improved, sex-dependent training procedures for awake mouse MRI. We recorded heart rate, monitored behavioral responses (body weight and fecal boli weight) and fecal corticosterone levels (FCM) as indicators of wellbeing and stress during a 14-day progressive habituation protocol. In addition, we also assessed discomfort levels and anxiety using the mouse grimace scale (MGS) and light/dark test (LDT), respectively. All scores were compared between both groups. We found that heart rate was significantly decreased after 10 and 11 days of training for both males and females, respectively. However, the specific time course for this decrease was significantly different between males and females, and females exhibited higher anxiety levels during habituation and 14 days after habituation than males. Lastly, we also found that mean FCM levels for both groups were decreased after 11 days of MRI habituation. The present work shows that mice can be successfully trained for extended MRI sessions which is necessary for many (particularly non-fMRI) studies. Importantly, we find that males and females differ in their response to awake MRI habituation, which should be considered in future awake MRI studies that aim to include male and female mice.
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BACKGROUND AND PURPOSE: Autism spectrum disorders (ASD) are heterogeneous neurodevelopmental disorders with considerably increased risk in male infants born preterm and with neonatal infection. Here, we investigated the role of postnatal immune activation on hippocampal synaptopathology by targeting Reelin+ cells in mice with ASD-like behaviours. EXPERIMENTAL APPROACH: C57/Bl6 mouse pups of both sexes received lipopolysaccharide (LPS, 1 mg·kg-1 ) on postnatal day (P) 5. At P45, animal behaviour was examined by marble burying and sociability test, followed by ex vivo brain MRI diffusion kurtosis imaging (DKI). Hippocampal synaptogenesis, number and morphology of Reelin+ cells, and mRNA expression of trans-synaptic genes, including neurexin-3, neuroligin-1, and cell-adhesion molecule nectin-1, were analysed at P12 and P45. KEY RESULTS: Social withdrawal and increased stereotypic activities in males were related to increased mean diffusivity on MRI-DKI and overgrowth in hippocampus together with retention of long-thin immature synapses on apical dendrites, decreased volume and number of Reelin+ cells as well as reduced expression of trans-synaptic and cell-adhesion molecules. CONCLUSION AND IMPLICATIONS: The study provides new insights into sex-dependent mechanisms that may underlie ASD-like behaviour in males following postnatal immune activation. We identify GABAergic interneurons as core components of dysmaturation of excitatory synapses in the hippocampus following postnatal infection and provide cellular and molecular substrates for the MRI findings with translational value.