Unveiling the Role of Endoplasmic Reticulum Stress Pathways in Canine Demodicosis.

Canine demodicosis is a prevalent skin disease caused by overpopulation of a commensal species of Demodex mite, yet its precise cause remains unknown. Research suggests that T-cell exhaustion, increased immunosuppressive cytokines, induction of regulatory T cells and increased expression of immune checkpoint inhibitors may contribute to its pathogenesis. This study aimed to gain a deeper understanding of the molecular changes occurring in canine demodicosis using mass spectrometry and pathway enrichment analysis. The results indicate that endoplasmic reticulum stress promotes canine demodicosis through regulation of three linked signalling pathways: eIF2, mTOR, and eIF4 and p70S6K. These pathways are involved in the modulation of Toll-like receptors, most notably TLR2, and have been shown to play a role in the pathogenesis of skin diseases in both dogs and humans. Moreover, these pathways are also implicated in the promotion of immunosuppressive M2 phenotype macrophages. Immunohistochemical analysis, utilising common markers of dendritic cells and macrophages, verified the presence of M2 macrophages in canine demodicosis. The proteomic analysis also identified immunological disease, organismal injury and abnormalities and inflammatory response as the most significant underlying diseases and disorders associated with canine demodicosis. This study demonstrates that Demodex mites, through ER stress, unfolded protein response and M2 macrophages contribute to an immunosuppressive microenvironment, thereby assisting in their proliferation.

Evaluation of micromilling/conventional isotope ratio mass spectrometry and secondary ion mass spectrometry of δ18 O values in fish otoliths for sclerochronology.

RATIONALE: Stable oxygen isotope ratios (δ18 O values) measured in fish otoliths can provide valuable detailed information on fish life history, fish age determination, and ocean thermography. Traditionally, otoliths are sampled by micromilling followed by isotope ratio mass spectrometry (IRMS), but direct analysis by secondary ion mass spectrometry (SIMS) is becoming more common. However, these two methods have not been compared to determine which, if either, is better for fish age validation studies. Hence, the goals were to: (1) determine if the δ18 O signatures from the two different methods are similar, (2) determine which method is better for fish age validation studies, and (3) examine biogeographic and migration history. METHODS: Both analytical techniques, micromilling/IRMS and SIMS, were used to measure δ18 O values in six Pacific cod (Gadus macrocephalus) otoliths. A series of measurements was made from the center of each otolith to its edge to develop a life-history δ18 O signature for each fish. RESULTS: The sampling resolution of SIMS analyses was 2-3 times greater than that obtained by micromilling/IRMS. We found an offset between SIMS and micromilling/IRMS δ18 O values, about 0.5‰ on average, with SIMS yielding lower values. However, the δ18 O patterns from both methods (i.e., the number of δ18 O maxima) correspond to the estimated age determined by otolith growth-zone counts, validating fish age determination methods. CONCLUSIONS: Both techniques resolved δ18 O life-history signatures and showed patterns consistent with seasonal variation in temperatures and changes due to fish migration. When otoliths are large, micromilling/IRMS can provide adequate resolution for fish age validation. However, SIMS is the better option if greater sampling resolution is required, such as when otoliths are small or specimens are longer lived and have compact growth zones.

Clinical characteristics and preventable acute care spending among a high cost inpatient population.

BACKGROUND: A small proportion of patients account for the majority of health care spending. The objectives of this study were to explore the clinical characteristics, patterns of health care use, and the proportion of acute care spending deemed potentially preventable among high cost inpatients within a Canadian acute-care hospital. METHODS: We identified all individuals within the Ottawa Hospital with one or more inpatient hospitalization between April 1, 2010 and March 31, 2011. Clinical characteristics and frequency of hospital encounters were captured in the information systems of the Ottawa Hospital Data Warehouse. Direct inpatient costs for each encounter were summed using case costing information and those in the upper first and fifth percentiles of the cumulative direct cost distribution were defined as extremely high cost and high cost respectively. We quantified preventable acute care spending as hospitalizations for ambulatory care sensitive conditions (ACSC) and spending attributable to difficulty discharging patients as measured by alternate level of care (ALC) status. RESULTS: During the study period, 36,892 patients had 44,066 hospitalizations. High cost patients (n = 1,844) accounted for 38 % of total inpatient spending ($122 million) and were older, more likely to be male, and had higher levels of co-morbidity compared to non-high cost patients. In over half of the high cost cohort (54 %), costs were accumulated from a single hospitalization. The majority of costs were related to nursing care and intensive care unit spending. High cost patients were more likely to have an encounter deemed to be ambulatory care sensitive compared to non-high cost inpatients (6.0 versus 2.8 %, p < 0.001). A greater proportion of inpatient spending was attributable to ALC days for high cost versus non-high cost patients (9.1 versus 4.9 %, p < 0.001). CONCLUSIONS: Within a population of high cost inpatients, the majority of costs are attributed to a single, non-preventable, acute care episode. However, there are likely opportunities to improve hospital efficiency by focusing on different approaches to community based care directed towards specific populations.

Influence of educational, audit and feedback, system based, and incentive and penalty interventions to reduce laboratory test utilization: a systematic review.

Laboratory and radiographic tests are often ordered unnecessarily. This excess testing has financial costs and is a burden on patients. We performed a systematic review to determine the effectiveness interventions to reduce test utilization by physicians. The MEDLINE and EMBASE databases were searched for the years 1946 through to September 2013 for English articles that had themes of test utilization and cost containment or optimization. Bibliographies of included papers were scanned to identify other potentially relevant studies. Our search resulted in 3236 articles of which 109 met the inclusion criteria of having an intervention aimed at reducing test utilization with results that could be expressed as a percent reduction in test use relative to the comparator. Each intervention was categorized into one or more non-exclusive category of education, audit and feedback, system based, or incentive or penalty. A rating of study quality was also performed. The percent reductions in test use ranged from a 99.7% reduction to a 27.7% increase in test use. Each category of intervention was effective in reducing test utilization. Heterogeneity between interventions, poor study quality, and limited time horizons makes generalizations difficult and calls into question the validity of results. Very few studies measure any patient safety or quality of care outcomes affected by reduced test use. There are numerous studies that use low investment strategies to reduce test utilization with one time changes in the ordering system. These low investment strategies are the most promising for achievable and durable reductions in inappropriate test use.

Tree-ring stable isotopes record the impact of a foliar fungal pathogen on CO(2) assimilation and growth in Douglas-fir.

Swiss needle cast (SNC) is a fungal disease of Douglas-fir (Pseudotsuga menziesii) that has recently become prevalent in coastal areas of the Pacific Northwest. We used growth measurements and stable isotopes of carbon and oxygen in tree-rings of Douglas-fir and a non-susceptible reference species (western hemlock, Tsuga heterophylla) to evaluate their use as proxies for variation in past SNC infection, particularly in relation to potential explanatory climate factors. We sampled trees from an Oregon site where a fungicide trial took place from 1996 to 2000, which enabled the comparison of stable isotope values between trees with and without disease. Carbon stable isotope discrimination (Δ(13)C) of treated Douglas-fir tree-rings was greater than that of untreated Douglas-fir tree-rings during the fungicide treatment period. Both annual growth and tree-ring Δ(13)C increased with treatment such that treated Douglas-fir had values similar to co-occurring western hemlock during the treatment period. There was no difference in the tree-ring oxygen stable isotope ratio between treated and untreated Douglas-fir. Tree-ring Δ(13)C of diseased Douglas-fir was negatively correlated with relative humidity during the two previous summers, consistent with increased leaf colonization by SNC under high humidity conditions that leads to greater disease severity in following years.

Determination of the safety and efficacy of therapeutic neutralization of tumor necrosis factor-α (TNF-α) using AZD9773, an anti-TNF-α immune Fab, in murine CLP sepsis.

OBJECTIVE AND DESIGN: TNF-α neutralization is associated with increased mortality in mouse cecal ligation puncture (CLP) models. AZD9773 is an ovine polyclonal human TNF-α immune Fab, with pharmacological properties that differ from previously studied anti-TNF-α agents. We explored the safety and efficacy of therapeutically administered AZD9773 in mouse CLP sepsis. METHODS: A moderate/severe-grade CLP model resulting in 20-30 % 5-day survival and a mild-grade CLP model resulting in ~70 % 5-day survival were established in human TNF-α transgene/murine TNF null (Tg1278/-/-) mice. TREATMENT: Mice received saline resuscitation and imipenem administration every 12 h (0-72 h post-CLP). AZD9773 (or DigiFab control) was dosed 24, 36, 48 and 60 h post-CLP. RESULTS: Therapeutic dosing of AZD9773 in moderate/severe-grade CLP resulted in significantly increased survival (>70 %) compared with DigiFab (27 %, P < 0.05). Therapeutic dosing of AZD9773 in mild-grade CLP did not significantly affect survival outcome compared with DigiFab or imipenem alone (~60-70 % survival). CONCLUSIONS: These data demonstrate that TNF-α neutralization can improve survival in moderate/severe CLP sepsis. TNF-α suppression in mild-grade models was not associated with survival benefit and did not increase 5-day mortality. These findings suggest that therapeutic benefit following TNF-α attenuation in models of sepsis may depend on model severity.

Including Patient Voices in Continuing Medical Education: One Provider's Experience.

In 2021, UpToDate began offering continuing medical education (CME) planned and delivered by patients. The patient-authored medical topic reviews focus on lessons learned from interactions with the healthcare system and emphasise quality of life for those living with specific conditions. Having access to the patient voice at the point of care provides clinicians with a perspective that can improve patient-provider communication and promote shared decision-making. Participants who viewed the patient-authored topics were emailed a survey about the content; several responses indicated that the new topics were useful in clinical practice. While positive responses demonstrate that clinicians value the patient perspective, we also received replies from participants and from the patient authors themselves indicating there is more work to be done in developing patient-led CME. As more patients are invited to join the conversation, their expertise will be increasingly recognised as integral to CME.

Gene expression analysis of Canine Demodicosis; A milieu promoting immune tolerance.

Canine demodicosis is a common skin disease seen in companion animal practice that results from an overpopulation of the commensal Demodex mite species. Common predisposing factors to the development of canine demodicosis include immunosuppressive diseases, such as neoplasia and hypothyroidism, and administration of immunosuppressive therapies, such as corticosteroids. Despite this, the pathogenesis of development of canine demodicosis remains unclear. Previous studies have implicated a role for increased expression of toll like receptor 2 (TLR2), increased production of interleukin (IL)-10) and T cell exhaustion. Here, we investigate gene expression of formalin fixed paraffin embedded skin samples from twelve cases of canine demodicosis in comparison to twelve healthy controls, using a 770 gene panel (NanoString Canine IO Panel). Results show an increase in the T cell population, specifically Th1 and Treg cells in dogs with demodicosis. In addition, while there is an upregulation of immunosuppressive cytokines such as IL-10 and IL-13, there is also an upregulation of immune check point molecules including PD-1/PD-L1 and CTLA-4. These findings suggest that Demodex spp. mites are modulating the host immune system to their advantage through upregulation of several immune tolerance promoting pathways.

Impact of Lung Disease on COVID-19 Health Outcomes in People Living With HIV.

Introduction COVID-19 most commonly causes pulmonary/lung infection, and these pulmonary diseases can complicate HIV infection. Underlying pulmonary diseases in people living with HIV (PLWH) could affect health outcomes if infected with COVID-19. Therefore, this study was designed to determine the impact of pulmonary diseases on the health outcomes of PLWH that were infected with COVID-19. Materials and methods We conducted a retrospective study to assess the impact of superimposed COVID-19 infection on pre-existing lung pathologies in patients living with human immunodeficiency virus (HIV) infection using data from the Minnesota Fairview network from January 1, 2020 to December 31, 2022. Ordinal logistic regressions were used to determine the effect of lung comorbidities on COVID-19 severity, COVID-19-specific mortality, and all-cause mortality, adjusting for patient age and gender.  Results Two hundred sixteen PLWH tested positive for COVID-19. 24.54% of these patients had one or more pulmonary diseases, including asthma, chronic obstructive pulmonary disease (COPD), and other lung diseases (interstitial lung diseases and pulmonary hypertension). The severity of COVID-19 outcomes was evaluated by the ranking of patients' medical records of testing positive, admitted to the hospital, being admitted to the ICU, and death. COVID-19-specific and all-cause mortality were evaluated separately. PLWH with underlying asthma or COPD was not associated with increased all-cause or COVID-19-specific mortality. Interstitial lung disease or pulmonary hypertension was significantly associated with poor health outcomes for COVID-19-specific mortality and all-cause mortality (Fisher's Exact p-value <0.001), with ICU admissions accounting for the most impact. Using the multivariate models, interstitial lung disease and pulmonary hypertension was significantly associated with an increased risk of more severe COVID-19 outcomes and COVID-19-specific mortality (OR=6.6153, CI=2.5944, 17.0795, p-value < 0.001). Interstitial lung disease and pulmonary hypertension were also significantly associated with an increased risk of more severe COVID-19 outcomes and all-cause mortality (OR=​​5.0885, CI=2.0590, 12.5542, p-value < 0.001). Conclusions To mitigate the poor outcomes associated with interstitial lung diseases and pulmonary hypertension in PLWH due to COVID-19, healthcare providers must educate their patients about safety measures against the COVID-19 vaccine. They can also encourage the COVID-19 vaccine uptake among their eligible patients.

Relationship between blood mercury levels and components of male song in Nelson's sparrows (Ammodramus nelsoni).

Mercury (Hg) adversely affects the health and behavior of exposed wildlife; however, behavioral effects remain largely unknown. Changes in avian singing behavior may affect a male's fitness because song reveals male quality and thus influences female mate choice and male territory-holding ability. Nelson's sparrows (Ammodramus nelsoni) live exclusively on salt marshes and risk high levels of Hg exposure and bioaccumulation. We recorded songs of male Nelson's sparrows at two locations with different Hg exposure to determine if total blood Hg concentration was related to song characteristics, as previously reported for other species. Males with higher blood Hg levels sang at higher maximum tonal frequency, but blood Hg and site location did not influence low tonal frequency and bout duration, contrary to predictions based on other species. Within the contaminated site, Hg levels were related to bouts per minute and gap duration, such that males at that site sang faster songs. Hg influences hormones and alters brain development, raising questions about specific effects on the brains and singing behavior of male Nelson's sparrows.

A retrospective study of cases of canine demodicosis submitted to a commercial diagnostic laboratory servicing the United Kingdom and Ireland (2017-2018) part 2; Aerobic culture and antimicrobial susceptibility results.

Clinical diagnostic reports from 508 cases of canine demodicosis diagnosed either by histological or skin scraping analysis from a United Kingdom Accreditation Service (UKAS) accredited veterinary diagnostic laboratory servicing the United Kingdom (UK) and Ireland were evaluated. Of the 508 cases, 284 had skin swabs submitted for culture on the same day the skin biopsy and/or skin scraping were obtained. Dogs with juvenile-onset (JO) demodicosis represented 57.4% of these cases, whilst adult-onset (AO) cases comprised 42.6%. The data revealed that overgrowth of pathogenic bacteria was more common in AO demodicosis cases (75.2%) in comparison to the JO cases (57%). Adult-onset cases also had increased involvement of bacteria belonging to multiple genera and/or yeast (28.9%) in comparison to JO cases (18.4%). Pruritus was significantly associated with an overgrowth of Staphylococcus pseudintermedius (p < 0.001). Resistance to one or more antimicrobial classes was noted in S. pseudintermedius isolates from 56.3% of JO cases with 10.3% of these cases being classified as Multi-Drug Resistant (MDR). Similarly, 51.9% of S. pseudintermedius isolates from the AO cases were noted to be resistant to one or more antimicrobial class with 8.6% of these cases being considered MDR. Cephalosporins were the most frequently administered antimicrobial class noted in submission histories, followed by the penicillin and fluoroquinolone classes. Whilst our findings reveal a high prevalence of concurrent overgrowth of pathogenic bacteria warranting therapeutic intervention in canine demodicosis, the presence of resistance within isolates highlights the need for prudent selection and targeted use of antimicrobial therapy that encompass the key principles of antimicrobial stewardship.

A retrospective study of cases of canine demodicosis submitted to a commercial diagnostic laboratory servicing the United Kingdom and Ireland (2017-2018): Part 1 - Signalment, lesion distribution, treatments, and concurrent diseases.

Canine demodicosis, due to an overpopulation of Demodex spp. mites, remains one of the most common dermatological diseases encountered in small animal practice. The aims of this study were to interrogate submitted histories and diagnostic report results from a large cohort of dogs (n = 508) diagnosed with demodicosis either through histological analysis or the finding of Demodex spp. mites on skin scrapings by a UKAS accredited commercial laboratory servicing the United Kingdom (UK) and Ireland in the years 2017 and 2018. The main findings revealed that short-coated breeds were more likely to develop juvenile-onset (JO) demodicosis, whereas medium- and long-coated breeds were more likely to develop adult-onset (AO) disease. Pododemodicosis was reported more commonly in adult, long-coated breeds. Skin scrapings were positive in only 83.3% of samples that had a corresponding positive biopsy result; this finding highlights the necessity to perform further diagnostic tests if demodicosis remains clinically suspected despite a negative skin scraping result. Concurrent underlying diseases, potentially associated with immunosuppression, were reported in 42/221 (19%) of dogs with AO demodicosis. Serum allergy and Sarcoptes ELISA assays were positive in individual animals in both the JO and AO groups; the clinical significance of these latter findings requires careful interpretation in dogs with confirmed demodicosis.

Validation of a Liquid Biopsy Protocol for Canine BRAFV595E Variant Detection in Dog Urine and Its Evaluation as a Diagnostic Test Complementary to Cytology.

A significant proportion of canine urothelial carcinomas carry the driver valine to glutamic acid variation (V595E) in BRAF kinase. The detection of V595E may prove suitable to guide molecularly targeted therapies and support non-invasive diagnosis of the urogenital system by means of a liquid biopsy approach using urine. Three cohorts and a control group were included in this multi-step validation study which included setting up a digital PCR assay. This was followed by investigation of preanalytical factors and two alternative PCR techniques on a liquid biopsy protocol. Finally, a blind study using urine as diagnostic sample has been carried out to verify its suitability as diagnostic test to complement cytology. The digital PCR (dPCR) assay proved consistently specific, sensitive, and linear. Using the dPCR assay, the prevalence of V595E in 22 urothelial carcinomas was 90.9%. When compared with histopathology as gold standard in the blind-label cases, the diagnostic accuracy of using the canine BRAF (cBRAF) variation as a surrogate assay against the histologic diagnosis was 85.7% with 92.3% positive predictive value and 80.0% negative predictive value. In all the cases, in which both biopsy tissue and the associated urine were assayed, the findings matched completely. Finally, when combined with urine sediment cytology examination in blind-label cases with clinical suspicion of malignancy, the dPCR assay significantly improved the overall diagnostic accuracy. A liquid biopsy approach on urine using the digital PCR may be a valuable breakthrough in the diagnostic of urothelial carcinomas in dogs.

The induction of macrophage hemeoxygenase-1 is protective during acute kidney injury in aging mice.

Aging is thought to be associated with a higher susceptibility to renal ischemia-reperfusion injury (IRI). To study whether defective induction of hemeoxygenase-1 (HO-1, a protective and anti-inflammatory enzyme) might contribute to this, we found that while 12-month-old mice had similar baseline renal function and HO-1 expression, the induction of HO-1 usually seen in ischemia-reperfusion was reduced. This was also associated with worsened renal function and acute tubular necrosis in the aged compared with young mice. In the older mice, heme arginate (HA) induced HO-1 in the cortex and medulla, significantly improved renal function, and reduced tissue injury. Cellular HO-1 induction in the medulla in response to injury or HA treatment was found to be interstitial rather than epithelial, as evidenced by its colocalization with macrophage markers. In vitro, HA treatment of primary macrophages resulted in marked HO-1 induction without impairment of classical activation pathways. Macrophage depletion, caused by diphtheria toxin treatment of 12-month-old CD11b-DTR transgenic animals, resulted in the loss of interstitial HO-1-positive cells and reversal of the protective phenotype of HA treatment. Thus, failure of HO-1 induction following renal IRI worsens structural and functional injury in older mice and represents a therapeutic target in the elderly. Hence, HO-1-positive renal macrophages mediate HA-induced protection in IRI.

Proceedings of the 2010 National Toxicology Program Satellite Symposium.

The 2010 annual National Toxicology Program (NTP) Satellite Symposium, entitled "Pathology Potpourri," was held in Chicago, Illinois, in advance of the scientific symposium sponsored jointly by the Society of Toxicologic Pathology (STP) and the International Federation of Societies of Toxicologic Pathologists (IFSTP). The goal of the annual NTP Symposium is to present current diagnostic pathology or nomenclature issues to the toxicologic pathology community. This article presents summaries of the speakers' presentations, including diagnostic or nomenclature issues that were presented, along with select images that were used for voting or discussion. Some topics covered during the symposium included a comparison of rat and mouse hepatocholangiocarcinoma, a comparison of cholangiofibrosis and cholangiocarcinoma in rats, a mixed pancreatic neoplasm with acinar and islet cell components, an unusual preputial gland tumor, renal hyaline glomerulopathy in rats and mice, eosinophilic substance in the nasal septum of mice, INHAND nomenclature for proliferative and nonproliferative lesions of the CNS/PNS, retinal gliosis in a rat, fibroadnexal hamartoma in rats, intramural plaque in a mouse, a treatment-related chloracne-like lesion in mice, and an overview of mouse ovarian tumors.

Recommendations for pathology peer review.

Pathology peer review verifies and improves the accuracy and quality of pathology diagnoses and interpretations. Pathology peer review is recommended when important risk assessment or business decisions are based on nonclinical studies. For pathology peer review conducted before study completion, the peer-review pathologist reviews sufficient slides and pathology data to assist the study pathologist in refining pathology diagnoses and interpretations. Materials to be reviewed are selected by the peer-review pathologist. Consultations with additional experts or a formal (documented) pathology working group may be used to resolve discrepancies. The study pathologist is solely responsible for the content of the final pathology data and report, makes changes resulting from peer-review discussions, initiates the audit trail for microscopic observations after all changes resulting from peer-review have been made, and signs the final pathologist's report. The peer-review pathologist creates a signed peer-review memo describing the peer-review process and confirming that the study pathologist's report accurately and appropriately reflects the pathology data. The study pathologist also may sign a statement of consensus. It is not necessary to archive working notes created during the peer-review process.

CME in the Time of COVID-19: Educating Healthcare Professionals at the Point-of-care and Improving Performance Outcomes.

UpToDate® is a point-of-care clinical resource used by nearly 2 million clinicians worldwide. Users in the USA and other regions can obtain continuing education credits after researching clinical questions at the point-of-care. As part of the CME credit redemption process, participants provide feedback on the information researched during the UpToDate® learning activity. We examined the impact of UpToDate® searches on clinical decision-making related to COVID-19. Between January 1 and 31 August 2020, UpToDate® added more than 40 topic reviews on various aspects of COVID-19 diagnosis and management, and developed new methods for delivering COVID-19 content to our learners. During the observation period, participants accessed COVID-19-related topic reviews over 7.5 million times. Data collected from CME activity evaluations and user feedback suggested that the learning activity had a significant impact on clinical decision-making. Over 94% reported that they modified their management strategies as a result of using UpToDate®, and 97% reported that use of UpToDate® led to improvement in care. These findings support the benefit of self-directed, point-of-care learning activities on the clinical management of patients during a global pandemic.

The role of Northeast Pacific meltwater events in deglacial climate change.

Columbia River megafloods occurred repeatedly during the last deglaciation, but the impacts of this fresh water on Pacific hydrography are largely unknown. To reconstruct changes in ocean circulation during this period, we used a numerical model to simulate the flow trajectory of Columbia River megafloods and compiled records of sea surface temperature, paleo-salinity, and deep-water radiocarbon from marine sediment cores in the Northeast Pacific. The North Pacific sea surface cooled and freshened during the early deglacial (19.0-16.5 ka) and Younger Dryas (12.9-11.7 ka) intervals, coincident with the appearance of subsurface water masses depleted in radiocarbon relative to the sea surface. We infer that Pacific meltwater fluxes contributed to net Northern Hemisphere cooling prior to North Atlantic Heinrich Events, and again during the Younger Dryas stadial. Abrupt warming in the Northeast Pacific similarly contributed to hemispheric warming during the Bølling and Holocene transitions. These findings underscore the importance of changes in North Pacific freshwater fluxes and circulation in deglacial climate events.