Ocular biometry in rabbits using computed tomography.

OBJECTIVE: To describe a repeatable method of measuring ocular structures and to establish ocular biometry reference ranges in adult domestic rabbits (Oryctolagus cuniculus) without medical history or imaging findings consistent with ophthalmic disease using a 64-slice multidetector computed tomography scanner. PROCEDURE: In this retrospective and observational anatomic study, 100 eyes from 50 rabbits without medical history or imaging findings consistent with ophthalmic disease who received a head computed tomography scan were selected for measurement of globe length, width, and height using 3D multiplanar reconstruction. Lens width and length, the anteroposterior length of the anterior and vitreous chambers, and attenuation of the lens and vitreous chamber were collected. These parameters were compared against age, sex, weight, body condition, and ear conformation. RESULTS: A reference guide was established, with globe width being the largest dimension (18.03 ± 0.81 mm), followed by height (17.18 ± 0.69 mm) and then length (16.64 ± 0.66 mm). Increased weight was associated with an increase in globe height (p = 2.43 × 10-5 ), length (p = 1.63 × 10-4 ), and width (p = 7.0 × 10-3 ). Increased age was associated with increased lens attenuation (p = 1.28 × 10-7 ) and increased transverse lens width (p = 1.64 × 10-3 ). Inter- and intra-observer agreement was excellent. CONCLUSIONS: CT is a reliable modality for measurement of ocular biometry dimensions in rabbits. These reference values can be applied to aid in identifying diseases that affect the dimensions of the ocular structures in rabbits over 18 months of age.

Ocular mycobacterial lesions in cats.

Ocular mycobacterial infections are an under-recognized cause of morbidity in the domestic cat. This study aimed to explore the distribution, histopathological appearance, and severity of feline ocular mycobacterial lesions, and to characterize the immune cell population with immunohistochemistry. Routine histological staining with hematoxylin and eosin, and Masson's trichrome, was performed to identify ocular lesions and assign an inflammation score based on the number of cells present. Acid-fast bacilli were detected with Ziehl-Neelsen, and immunohistochemistry for ionized calcium-binding adaptor protein-1 (Iba1), calprotectin, cluster of differentiation 3 (CD3), and Pax5 was undertaken on formalin-fixed paraffin-embedded tissue samples from 24 cases of ocular mycobacteriosis. Posterior or panuveitis with concurrent retinitis was identified in 20/24 cases (83%), with retinal detachment in 16/20 (80%) of these cases. Choroidal lesions had the highest median inflammation score. Ziehl-Neelsen-positive organisms were detected in 20/24 cases (83%), with the highest prevalence of acid-fast bacilli detected in choroidal lesions (16/20, 80%). Lesions were typically granulomatous to pyogranulomatous, characterized by abundant numbers of Iba1-positive macrophages, followed by calprotectin-positive granulocytes and monocytes, fewer T cells, and rarer B cells. However, where iritis was identified, inflammation was typically lymphoplasmacytic (11/16 cases, 69%). Where diagnostic testing was performed, tuberculosis (ie, infection with Mycobacterium bovis, Mycobacterium microti, or a nonspeciated Mycobacterium tuberculosis-complex pathogen) was diagnosed in 20/22 cats (91%), with Mycobacterium lepraemurium infection identified in the other 2/22 cats (9%). These results suggest the choroid is the primary site of lesion development in most cases of feline ocular mycobacteriosis, and inflammatory changes are associated with the presence of mycobacteria localized to ocular tissues.

Clinical and CT sialography findings in 22 dogs with surgically confirmed sialoceles.

Sialoceles are an uncommon canine salivary gland disease, and complete surgical resection is important for a positive outcome. Radiographic sialography has been described as a diagnostic test for presurgical planning; however, superimposition artifacts may limit the diagnosis and detection of all affected glands. Computed tomographic (CT) sialography is a promising technique for delineating the salivary gland apparatus. The aims of this retrospective, observational study were to describe clinical and CT sialographic findings in a group of dogs with confirmed sialoceles, to determine the sensitivity of CT sialography for detecting affected salivary glands using surgery as the reference standard and to determine interobserver agreement for CT sialographic assessments. Dogs were included if they underwent a CT sialography study followed by surgical resection of the diseased gland(s) and histopathological analysis. Computed tomography sialography studies of dogs with surgically confirmed sialoceles (n = 22) were reviewed by a European College of Veterinary Diagnostic Imaging (ECVDI)-certified radiologist and an ECVDI resident. Interobserver agreement was calculated using Cohen's kappa statistics. CT sialography results were compared to surgical findings to determine sensitivity. Contrast leakage was detected in 12 of 22 dogs (54.5%), with intrasialocele leakage being most frequently observed (7/12, 58.3%). There was substantial agreement (κ = 0.70) between reviewers identifying diseased glands, substantial agreement (κ = 0.62) on the diagnostic quality, and no to slight agreement (к = 0.13) in the detection of contrast leakage. The overall sensitivity of CT sialography to detect surgically confirmed diseased glands was 66.7% (95% confidence interval: 48.8-80.8). In conclusion, these findings support the use of CT sialography as an adjunct diagnostic test for treatment planning in dogs with sialoceles.

Physician Social Media Abuse: What Would You Do?

Over the past decade, physician social media use has been made popular and evolved from simple Facebook pages to rapid, instant media sharing. New social media applications, such as Snapchat and Instagram, are finding welcome homes in personal cell phones of physicians. The purpose of this article is to determine patients' views of not only physician use of these apps but also how they would react if their physician abused this technology. As outlined in the article below, physician abuse of social media is growing-and not strictly confined to one demographic group of physicians. The results of this study show patient concern over physician use of social media and the potential for patient to take legal action against physicians over social media abuse. Future policy directions should aim to increase transparency of physician social media use with the goal of eliminating social media abuse.

Ground Same Intratransport Efficacy as Air for Acute Aortic Diseases.

INTRODUCTION: Patients with acute aortic diseases (AAoD) usually require transfer to tertiary centers for possible surgical care, for which intratransport management represents important continuing spectrum of care. There is little information comparing intratransport efficacy of air (ART) vs ground transport (GRT), nor how effectively they manage these patients' pain. Our study aims to compare how effective ART and GRT manage patients' intratransport HR, pressure. METHODS: Charts were reviewed of adult patients interhospital transferred to a quaternary academic center (UMMC) between 01/01/2011 and 09/30/2015. Outcomes were percentages of patients achieving target hemodynamic parameters, mortality. RESULTS: We analyzed 226 patients, 58 (26%) transported by Air and 102 (45%) type A dissection. Ground transport was associated with higher percentage of patients with target HR 60-80 bpm comparing to ART (58% vs 43%, 95% CI 0.3-0.99). Both ART and GRT were associated with similar frequencies of patients achieving target SBP and adequate pain control. Time intervals from transfer request to surgery, and mortality were similar for both types of transport. CONCLUSION: Ground transport teams were more successful at achieving predefined target heart rate than Air transport. Intra-transport management of other vital signs and pain were equally effectively between both Air and Ground transport.

The value of employees in a value-based care system.

To be successful under value-based payment, hospitals will require a healthy communication culture in which workers in the trenches are committed, engaged, and focused. In such a culture, employees should feel free to express their fears and doubts, accept and apply constructive feedback, and participate creatively. Mistakes should be seen not as cause for disciplinary action but as learning opportunities. New hires should have a willingness to continually learn and work independently, be unafraid of making mistakes, and be committed to adding value through the delivery of improved care and enhanced outcomes.

Social Media in Health Care: How Close Is Too Close?

Social media use is increasing personally and professionally across numerous industries worldwide. The purpose of this article is to explore the utilization of social media in the health care field; specifically, how the treatment of a physician's Facebook friends would differ from that of a patient the physician did not know prior to treatment. While there are several benefits that come with incorporating social media into health care, as well as into the physician-patient relationship, there are also immense risks. The present study surveyed physicians to assess their opinions on the boundaries of an appropriate patient-physician relationship on social media. Fifty-six of 70 physicians responded with their attitudes on the difference between adding a patient as a friend and adding a friend as a patient, as well as the difference in care between the two. The results of the study showed that most physicians would not be opposed to taking on their Facebook friends as patients; however, the care they provide could potentially be radically different between their Facebook friend and an average patient. This means that patients should take extra care before asking their Facebook physician friends for formal care when they are in need.

Quantitative analysis of the chromatin proteome in disease reveals remodeling principles and identifies high mobility group protein B2 as a regulator of hypertrophic growth.

A fundamental question in biology is how genome-wide changes in gene expression are enacted in response to a finite stimulus. Recent studies have mapped changes in nucleosome localization, determined the binding preferences for individual transcription factors, and shown that the genome adopts a nonrandom structure in vivo. What remains unclear is how global changes in the proteins bound to DNA alter chromatin structure and gene expression. We have addressed this question in the mouse heart, a system in which global gene expression and massive phenotypic changes occur without cardiac cell division, making the mechanisms of chromatin remodeling centrally important. To determine factors controlling genomic plasticity, we used mass spectrometry to measure chromatin-associated proteins. We have characterized the abundance of 305 chromatin-associated proteins in normal cells and measured changes in 108 proteins that accompany the progression of heart disease. These studies were conducted on a high mass accuracy instrument and confirmed in multiple biological replicates, facilitating statistical analysis and allowing us to interrogate the data bioinformatically for modules of proteins involved in similar processes. Our studies reveal general principles for global shifts in chromatin accessibility: altered linker to core histone ratio; differing abundance of chromatin structural proteins; and reprogrammed histone post-translational modifications. Using small interfering RNA-mediated loss-of-function in isolated cells, we demonstrate that the non-histone chromatin structural protein HMGB2 (but not HMGB1) suppresses pathologic cell growth in vivo and controls a gene expression program responsible for hypertrophic cell growth. Our findings reveal the basis for alterations in chromatin structure necessary for genome-wide changes in gene expression. These studies have fundamental implications for understanding how global chromatin remodeling occurs with specificity and accuracy, demonstrating that isoform-specific alterations in chromatin structural proteins can impart these features.

Chicken intestinal organoids: a novel method to measure the mode of action of feed additives.

There is a rapidly growing interest in how the avian intestine is affected by dietary components and feed additives. The paucity of physiologically relevant models has limited research in this field of poultry gut health and led to an over-reliance on the use of live birds for experiments. The development of complex 3D intestinal organoids or "mini-guts" has created ample opportunities for poultry research in this field. A major advantage of the floating chicken intestinal organoids is the combination of a complex cell system with an easily accessible apical-out orientation grown in a simple culture medium without an extracellular matrix. The objective was to investigate the impact of a commercial proprietary blend of organic acids and essential oils (OA+EO) on the innate immune responses and kinome of chicken intestinal organoids in a Salmonella challenge model. To mimic the in vivo prolonged exposure of the intestine to the product, the intestinal organoids were treated for 2 days with 0.5 or 0.25 mg/mL OA+EO and either uninfected or infected with Salmonella and bacterial load in the organoids was quantified at 3 hours post infection. The bacteria were also treated with OA+EO for 1 day prior to challenge of the organoids to mimic intestinal exposure. The treatment of the organoids with OA+EO resulted in a significant decrease in the bacterial load compared to untreated infected organoids. The expression of 88 innate immune genes was investigated using a high throughput qPCR array, measuring the expression of 88 innate immune genes. Salmonella invasion of the untreated intestinal organoids resulted in a significant increase in the expression of inflammatory cytokine and chemokines as well as genes involved in intracellular signaling. In contrast, when the organoids were treated with OA+EO and challenged with Salmonella, the inflammatory responses were significantly downregulated. The kinome array data suggested decreased phosphorylation elicited by the OA+EO with Salmonella in agreement with the gene expression data sets. This study demonstrates that the in vitro chicken intestinal organoids are a new tool to measure the effect of the feed additives in a bacterial challenge model by measuring innate immune and protein kinases responses.

Specialized compartments of cardiac nuclei exhibit distinct proteomic anatomy.

As host to the genome, the nucleus plays a critical role as modulator of cellular phenotype. To understand the totality of proteins that regulate this organelle, we used proteomics to characterize the components of the cardiac nucleus. Following purification, cardiac nuclei were fractionated into biologically relevant fractions including acid-soluble proteins, chromatin-bound molecules and nucleoplasmic proteins. These distinct subproteomes were characterized by liquid chromatography-tandem MS. We report a cardiac nuclear proteome of 1048 proteins--only 146 of which are shared between the distinct subcompartments of this organelle. Analysis of genomic loci encoding these molecules gives insights into local hotspots for nuclear protein regulation. High mass accuracy and complementary analytical techniques allowed the discrimination of distinct protein isoforms, including 54 total histone variants, 17 of which were distinguished by unique peptide sequences and four of which have never been detected at the protein level. These studies are the first unbiased analysis of cardiac nuclear subcompartments and provide a foundation for exploration of this organelle's proteomes during disease.

Community-Level Characteristics of Timely PCP Follow-Up Care and Post-Discharge ER Usage.

Achieving equitable access has been a central goal in healthcare for years; access by low-income Americans is a major concern for policymakers. We examined the differences in post-discharge primary care follow-up visits and 30-day post discharge ER visits across several characteristics. The results suggest that higher housing density, percent minority population, percent unemployed, and percent uninsured point to lower rates of PCP follow-up care and higher rates of post-discharge ER visits. These findings have implications for developing cost-effective programs targeting hospital to PCP communication, especially in densely populated areas.

Development of an enzyme-linked immunosorbent assay for the diagnosis of feline tuberculosis.

Cases of feline tuberculosis (TB) can be challenging to diagnose. Currently, this is achieved through a combination of mycobacterial culture, polymerase chain reaction (PCR), or interferon-gamma release assay (IGRA); however, these each have limitations. There is limited data regarding the use of humoral immunodiagnostics for TB in cats. Therefore, we sought to develop an enzyme-linked immunosorbent assay (ELISA) to further facilitate the diagnosis of feline TB. A comparative PPD (purified protein derivative) antibody ELISA was optimised for use on serum and plasma, and was tested against samples from 14 cats with culture-confirmed TB and 24 uninfected controls. Selection of an appropriate positive cut-off value based on receiver-operator characteristic curve analysis gave test sensitivity of 64.3 % and specificity of 100 %. When tested on further samples from cats with strongly suspected mycobacteriosis, 32.9 % (23/70) were antibody positive. Notably, positive results were recorded in cats that failed to respond to the IGRA, and in one PCR and IGRA negative cat. No positive responses were identified in cats with non-tuberculous mycobacterial infections, or with non-mycobacterial diseases (n = 12). Therefore, antibody-based diagnostics may be useful adjunctive tests for cases of TB missed by the IGRA, helping protect both feline and, in turn, human health.

Disentangling the innate immune responses of intestinal epithelial cells and lamina propria cells to Salmonella Typhimurium infection in chickens.

Salmonella enterica serovar Typhimurium (STm) is a major foodborne pathogen and poultry are a key reservoir of human infections. To understand the host responses to early stages of Salmonella infection in poultry, we infected 2D and 3D enteroids, the latter of which contains leukocytes, neurons, and mesenchymal cells that are characteristic of the lamina propria. We infected these enteroids with wild-type (WT STm), a non-invasive mutant lacking the prgH gene (ΔprgH STm), or treated them with STm lipopolysaccharide (LPS) and analyzed the expression of innate immune related genes by qPCR at 4 and 8 h. The localization of the tight junction protein, ZO-1, expression was disrupted in WT STm infected enteroids but not ΔprgH STm or LPS treated enteroids, suggesting a loss of epithelial barrier integrity. The innate immune response to LPS was more pronounced in 2D enteroids compared to 3D enteroids and by 8 hpi, the response in 3D enteroids was almost negligible. However, when STm adhered to or invaded the enteroids, both 2D and 3D enteroids exhibited an upregulation of inflammatory responses. The presence of lamina propria cells in 3D enteroids resulted in the unique expression of genes associated with immune functions involved in regulating inflammation. Moreover, 2D and 3D enteroids showed temporal differences in response to bacterial invasion or adherence. At 8 hpi, innate responses in 3D but not 2D enteroids continued to increase after infection with WT STm, whereas the responses to the non-invasive strain decreased at 8 hpi in both 2D and 3D enteroids. In conclusion, STm infection of chicken enteroids recapitulated several observations from in vivo studies of Salmonella-infected chickens, including altered epithelial barrier integrity based on ZO-1 expression and inflammatory responses. Our findings provide evidence that Salmonella-infected enteroids serve as effective models for investigating host-pathogen interactions and exploring the molecular mechanisms of microbial virulence although the 3D model mimics the host more accurately due to the presence of a lamina propria.

Features of endogenous cardiomyocyte chromatin revealed by super-resolution STED microscopy.

Despite the extensive knowledge of the functional unit of chromatin-the nucleosome-for which structural information exists at the atomic level, little is known about the endogenous structure of eukaryotic genomes. Chromosomal capture techniques and genome-wide chromatin immunoprecipitation and next generation sequencing have provided complementary insight into global features of chromatin structure, but these methods do not directly measure structural features of the genome in situ. This lack of insight is particularly troublesome in terminally differentiated cells which must reorganize their genomes for large scale gene expression changes in the absence of cell division. For example, cardiomyocytes, which are fully committed and reside in interphase, are capable of massive gene expression changes in response to physiological stimuli, but the global changes in chromatin structure that enable such transcriptional changes are unknown. The present study addressed this problem utilizing super-resolution stimulated emission depletion (STED) microscopy to directly measure chromatin features in mammalian cells. We demonstrate that immunolabeling of histone H3 coupled with STED imaging reveals chromatin domains on a scale of 40-70 nm, several folds better than the resolution of conventional confocal microscopy. An analytical workflow is established to detect changes in chromatin structure following acute stimuli and used to investigate rearrangements in cardiomyocyte genomes following agonists that induce cellular hypertrophy. This approach is readily adaptable to investigation of other nuclear features using a similar antibody-based labeling technique and enables direct measurements of chromatin domain changes in response to physiological stimuli.

The association between hospital care transition planning and timely primary care follow-up.

PURPOSE: To determine whether Hospital Consumer Assessment of Healthcare Providers and Systems (HCAHPS) Care Transitions (CTM-3) Scores were associated with timely (14-day) primary care provider (PCP) follow-up visits, and to look for disparities across various types of urban and rural hospitals. METHODS: Data were obtained for 3,299 hospitals: 2,000 urban, 544 micropolitan prospective payment system (PPS), 109 micropolitan critical access hospital (CAH), 252 noncore rural PPS, and 394 noncore rural CAH. HCAPPS data were drawn from CMS Hospital Compare (2015). The dependent variable, 14-day PCP follow-up rate for each hospital, was drawn from the 2015 Dartmouth Atlas. FINDINGS: In analysis adjusting only for hospital characteristics, higher CTM-3 scores were positively associated with PCP follow-up; however, the relationship was no longer significant after controlling for area-level (contextual) measures, such as percent minority population, percent unemployed, and percent uninsured. In the fully adjusted model, rates of PCP follow-up were significantly higher for micropolitan PPS, noncore PPS, and noncore CAH hospitals than for urban hospitals. CONCLUSIONS: In fully adjusted analysis, the lack of significance between CTM-3 scores and PCP follow-up suggests that community characteristics facilitate or impede timely PCP follow-up to an extent that may overshadow in-hospital efforts. Disparities between CAHs and rural PPS hospitals may be due to differing enrollments in quality incentive plans; future research is needed to address this issue. Compounding this issue, the strong negative relationship between percent Medicaid reimbursement (payor mix) and PCP follow-up suggests possible disparities for safety net hospitals.

Ocular Tuberculosis: More than 'Of Mice and Men'.

Tuberculosis (TB), caused by infection with members of the Mycobacterium tuberculosis-complex, is one of the oldest known infectious disease entities, resulting in the death of millions of humans each year. It also results in a substantial degree of morbidity and mortality in animal species. Extrapulmonary TB is well recognized in humans, and the eye is one site that can be affected. Studies seeking to understand ocular TB have often relied on animal models; however, these have their limitations and may not truly reflect what happens in humans. We wish to raise awareness among ophthalmologists and vision scientists of naturally occurring cases of ocular TB in animals, namely cattle and domestic cats, and the possibilities of gaining further understanding of this presentation of TB by adopting a collaborative approach. This will hopefully improve outcomes for both human and animal patients.

Recognition of recombinant interferon-gamma from Felidae species by anti-cat antibodies.

Mycobacterial infections cause a reasonable burden of morbidity and mortality in global feline populations, many of which are 'Vulnerable' or 'Endangered'. Identifying these infections may facilitate efforts to protect these animals. An interferon-gamma (IFNγ) release assay (IGRA) to diagnose mycobacteriosis in domestic cats has been adapted for use in lions; however, the development of species-specific antibodies may be laborious. Therefore, we investigated whether anti-cat IFNγ antibodies can bind to recombinant IFNγ (rIFNγ) from other Felidae species, permitting use of the feline IGRA in a wider range of felids. Unique Felidae IFNγ protein sequences and their corresponding coding nucleotide sequence were identified from online databases; plasmids with an IFNγ-gene insert were synthesised to transform E. coli-DH5α and subsequently transfect HEK 293 T cells to secrete rIFNγ. Enzyme-linked immunosorbent assay using a commercial anti-cat IFNγ kit was performed to detect rIFNγ from Felidae, the domestic dog and cattle. Five unique rIFNγ Felidae proteins were synthesised; anti-cat IFNγ antibodies were able to bind to all five proteins, while cross-reactivity with canine and bovine rIFNγ was negligible. This suggests that anti-cat IFNγ antibodies are sufficient for detection of IFNγ across other Felidae species, namely the lion, tiger, cheetah, cougar, Iberian lynx and the Canadian lynx.

Serial Interferon-Gamma Release Assay (IGRA) Testing to Monitor Treatment Responses in Cases of Feline Mycobacteriosis.

The interferon-gamma release assay (IGRA) is used to diagnose cases of feline mycobacteriosis, but the use of serial testing to monitor treatment responses has not been evaluated in this species. From a population of cats that underwent IGRA testing for diagnostic investigation, individuals were identified with a pre- and end-of-treatment IGRA that passed control thresholds. The number of cats which reverted to negative at the end-of-treatment IGRA, changes in paired antigen-specific optical density (OD) values and differences in the pre-treatment antigen-specific OD values for those which underwent reversion were compared. Factors to explain reversion or recurrence of disease post-treatment were explored. Four of 18 cats (22%) reverted to negativity at the point of clinical resolution (p = 0.33), there was no difference in paired antigen-specific OD values (p ≥ 0.12), and cats that reverted did not have a lower baseline OD value (p = 0.63). No statistically significant factors were identified to predict reversion (p ≥ 0.08). Remaining positive at the end of treatment IGRA was not associated with recurrence of disease post-treatment (p = 0.34). Overall, these data suggest there is limited value in the use of the IGRA to monitor treatment responses in cats.

Diagnostic accuracy of the interferon-gamma release assay (IGRA) for cases of feline mycobacteriosis.

The aim of this study was to evaluate the sensitivity and specificity of the interferon-gamma release assay (IGRA) for diagnosing infections with members of the Mycobacterium (M.) tuberculosis-complex (MTBC) and non-tuberculous mycobacteria (NTM) in domestic cats, and to generate defined feline-specific cut-off values using receiver operating characteristic (ROC) curve analysis to improve test performance. Records of 594 cats that had been tested by IGRA were explored to identify individuals that had a culture and/or polymerase chain reaction (PCR)-confirmed case of mycobacterial disease, and those that had a final diagnosis of non-mycobacterial disease. A total of 117 cats - 80 with mycobacterial disease and 37 diagnosed with a condition other than mycobacteriosis - were identified for further detailed analysis. This population was used to estimate test sensitivity and specificity, as well as likelihood ratios for the IGRA to correctly identify a cat with or without mycobacterial disease. Agreement between IGRA results and culture/PCR using current and proposed new cut-off values was also determined. ROC analysis of defined confirmed infected and non-mycobacterial disease control cats allowed an adjustment of current test cut-offs that increased the overall test sensitivity for MTBC infections from 83.1 % (95 % confidence interval [CI]: 71.5-90.5 %) to 90.2 % (95 % CI: 80.2-95.4%), and M. bovis infection from 43 % (95 % CI: 28.2-60.7%) to 68 % (95 % CI: 51.4-82.1%) while maintaining high test specificity (100 % in both cases). Overall agreement between IGRA results and culture/PCR, while recognising that neither culture nor PCR tests have perfect sensitivity, improved from weak (κ = 0.57) to moderate (κ = 0.71) using new proposed IGRA test cut-off values. Application of these results, based upon the statistical analysis of accumulated test data, can improve the diagnostic performance of the feline IGRA, particularly for identifying infections with M. bovis, without compromising specificity.