Unique Presentations of Burkholderia gladioli Infections in Several Strains of Immunocompromised Mice.

Four strains of experimentally naïve mice (NOD.Cg-Prkdcscid Il2rgtm1Wjl/SzJ [NSG], NOD.Cg-Rag1tm1Mom Il2rgtm1Wjl/SzJ[NRG], B6.129S(Cg)-Stat1tm1Dlv/J [STAT1-/-], and B6.129S7-Ifngr1tm1Agt/J [IFNγR-/-] housed in a barrier facility developedunusual and seemingly unrelated clinical signs. Young NSG/NRG mice (n = 49, mean age = 4 ± 0.4 mo) exhibited nonspecificclinical signs of moderate-to-severe lethargy, hunched posture, decreased body condition, and pallor. In contrast to the NSG/NRGmice, the STAT1-/- and IFNγR-/-mice (n = 5) developed large subcutaneous abscesses on the head and neck. These micewere euthanized, and samples were collected for culture. NSG/NRG mice had moderate-markedly enlarged livers (20 of49, 40%) and spleens (17 of 49, 35%). The livers contained multiple, variably-sized, tan regions throughout all lobes. Histologyrevealed necrotizing hepatitis (13 of 17, 77%), splenic and hepatic extramedullary hematopoiesis (17 of 17, 100%), glomerularhistiocytosis (6 of 17, 35%), and metritis (6 of 11, 55%) with perivascular inflammation, suggesting hematogenous spreadDifferentials for these lesions included mouse hepatitis virus, ectromelia virus, Pseudomonas aeruginosa, Salmonella spp.,and Clostridium piliforme. Burkholderia gladioli was cultured from liver lesions and subcutaneous abscesses and confirmedwith 16S ribosomal RNA sequencing. After completing systematic testing of the environment, failure of the water autoclavecycle was suspected as the cause of the outbreak. To address the situation, individually ventilated racks were sanitized andnew breeders were purchased; these actions dramatically reduced B. gladioli infections. The current literature contains fewreports of B. gladioli infections in immunocompromised mice, and its typical presentation is torticollis and rolling. B. gladioliinfection is a potential differential for subcutaneous abscesses, hepatitis, and splenomegaly in immunocompromised mice.Careful monitoring of sterilization techniques is essential to prevent such infections in a barrier facility.

Connection and conflict: influence of the hidden curriculum on veterinary residents' professional identities within the specialty of laboratory animal medicine.

OBJECTIVE: To explore the role of the hidden curriculum in residents' development of professional identity during postgraduate training in laboratory animal medicine. SAMPLE: 24 residents enrolled in 1 of 7 laboratory animal medicine training programs in the eastern US. PROCEDURE: 24 qualitative, semistructured interviews were conducted and recorded. Deidentified transcriptions were analyzed by each author using open and axial coding. Constant comparative methodology was used to develop themes and subthemes. Member checks were performed to verify trustability of the conclusions drawn. RESULTS: 3 themes and their related subthemes emerged from the qualitative analysis: 1) building relationships through competent communication (building rapport, practicing clinical empathy, overcoming language barriers, communicating in the "authorized" way, and navigating email limitations), 2) tension within the process of identity formation (acting as the middleman among stakeholders, overcoming the stigma of the policing role, experiencing a lack of power to impact change, and managing a culture of conditional value of veterinary knowledge), and 3) outlets for tension in identity formation (reliance on residency mates, limitations of venting). EDUCATIONAL RELEVANCE: Our findings suggest that residents are navigating professional identity formation under challenging circumstances that include conflicting stakeholder needs, conditional value of veterinary knowledge, and lack of power to influence change. Residents have limited outlets for relieving the discord between their ideal professional role and their lived experiences. These results provide an important background for refining curricula and creating effective support systems for residents.

Antibody Titers and Seroconversion Kinetics of Outbred Swiss and Heterozygous Nude Soiled-bedding Sentinels for Murine Norovirus and Mouse Hepatitis Virus.

Sentinel animals remain a common means of evaluating rodent health in research colonies. An evaluation of our sentinel program revealed that using Crl:CD1(ICR)-Elite (CD1-E) mice was expensive, occasionally disrupted by limited supply, and minimally responsive to the 3Rs. This evaluation prompted us to explore the use of CRL:NU-Foxn1nu/+ (Het-nude) mice as soiled-bedding sentinel (SBS) animals. Het-nude mice are a byproduct of breeding outbred athymic nude mice and are reared in isolators, with similar health status as CD1-E. Het-nude mice have a thymus, but may have smaller thymic size and fewer bone marrow stem cells than do wildtype controls, suggesting that Het-nude mice might not be immunologically normal. This study compared the antibody titer and seroconversion kinetics of Het-nude and CD1-E SBS to murine norovirus (MNV) and mouse hepatitis virus (MHV). Het-nude and CD1-E female SBS (n = 22 mice of each stock) were housed continuously on soiled bedding collected from MNV-positive or MNV- and MHV-positive colonies at cage changes. Blood was collected for serology at 3, 9 and 12 to 19 wk after the start of soiled bedding exposure. Antibody titers to MNV or MHV did not differ significantly between Het-nude and CD1-E mice. A significant relationship was found between weeks of exposure and titer levels with an increase in titer over the testing period. This study supports the possible use of Het-nude mice as SBS, given that their antibody responses to MNV and MHV are equivalent to those of CD1-E mice.

Role of the δ-Opioid Receptor in 2 Murine Models of Colitis.

Crohn disease and ulcerative colitis, collectively referred to as inflammatory bowel disease (IBD), are chronic inflammatory disorders of the gastrointestinal tract. Currently, the etiology of IBD is unknown, and immunosuppressive therapies have become the standard of care to reduce the inflammation; however, these agents only induce remission 50% of the time in patients and can have serious side effects. Recently, endogenous opioids and opioid receptors have been shown to play a role in the mediation of inflammation. In addition, opioid receptor blockade with a nonselective antagonist, naltrexone, has been shown to reduce colitis in both murine models and human subjects. The goal of the current study was to determine if the antiinflammatory effects of naltrexone are mediated through the delta (δ) opioid receptor. Male C57BL/6NCrl (6 to 8 wk.; n = 110) and female BALB/cAnNCrl (6-8 wk.; n = 91) mice were studied using 2 animal models of chemically induced colitis: dextran sodium sulfate (DSS) and 2, 4, 6-trinitrobenzenesulfonic acid (TNBS). The selective δ-receptor antagonists naltrindole and 7-benzylidenenaltrexone were administered to examine the role of the δ-opioid receptor in colonic inflammation. The quantitative measurement of colitis activity, colon weight and length, Hct, WBC count, and gross and microscopic aberrations were analyzed. Administration of naltrexone in the DSS colitis model significantly improved overall disease activity indices on day 5 of therapy. The use of δ-antagonists and naltrexone had limited to no effect on TNBS colitis. Similar findings were obtained by using the DSS colitis model. Based on the current findings, the authors conclude that naltrexone therapy has limited effect on the improvement of colitis in 2 murine models; however, the δ-opioid receptor was not responsible for mediating the effects.

Benefits and Challenges of Developing a Customized Rubric for Curricular Review of a Residency Program in Laboratory Animal Medicine.

Rigorous curricular review of post-graduate veterinary medical residency programs is in the best interest of program directors in light of the requirements and needs of specialty colleges, graduate school administrations, and other stakeholders including prospective students and employers. Although minimum standards for training are typically provided by specialty colleges, mechanisms for evaluation are left to the discretion of program directors. The paucity of information available describing best practices for curricular assessment of veterinary medical specialty training programs makes resources from other medical fields essential to informing the assessment process. Here we describe the development of a rubric used to evaluate courses in a 3-year American College of Laboratory Animal Medicine (ACLAM)-recognized residency training program culminating in a Master of Science degree. This rubric, based on examples from medical education and other fields of graduate study, provided transparent criteria for evaluation that were consistent with stakeholder needs and institutional initiatives. However, its use caused delays in the curricular review process as two significant obstacles to refinement were brought to light: variation in formal education in curriculum design and significant differences in teaching philosophies among faculty. The evaluation process was able to move forward after institutional resources were used to provide faculty development in curriculum design. The use of a customized rubric is recommended as a best practice for curricular refinement for residency programs because it results in transparency of the review process and can reveal obstacles to change that would otherwise remain unaddressed.

Presumptive Spontaneous Lysosomal Storage-Like Disease in a Crl:CD1(ICR) Mouse.

A clinically unremarkable 4.5-mo-old female Crl:CD1(ICR) VAF/Elite mouse was euthanized for scheduled sentinel processing. Gross necropsy findings included significant hepatosplenomegaly and visceral lymphadenomegaly, resulting in a preliminary gross diagnosis of lymphoma. Histology revealed florid accumulations of large, 'foamy' macrophages present in the bone marrow, small intestines, and viscera including liver, spleen, lymph nodes, thymus, uterus, and ovaries. The cytoplasm of these cells was abundant, stained pale blue with Wright-Giemsa and was periodic acid-Schiff positive. Given these characteristic gross and histologic findings, a spontaneous lysosomal storage-like disease was diagnosed in this mouse. Cholesterol ester storage disease is likely, because of the visceral involvement with sparing of the CNS, but could not be diagnosed definitively. To our knowledge, this report is the first to describe a case of spontaneous lysosomal storage disease in an outbred mouse of the CD1(ICR) background.

Time course of peri-implant bone regeneration around loaded and unloaded implants in a rat model.

The time-course of cancellous bone regeneration surrounding mechanically loaded implants affects implant fixation, and is relevant to determining optimal rehabilitation protocols following orthopaedic surgeries. We investigated the influence of controlled mechanical loading of titanium-coated polyether-ether ketone (PEEK) implants on osseointegration using time-lapsed, non-invasive, in vivo micro-computed tomography (micro-CT) scans. Implants were inserted into proximal tibial metaphyses of both limbs of eight female Sprague-Dawley rats. External cyclic loading (60 or 100 µm displacement, 1 Hz, 60 s) was applied every other day for 14 days to one implant in each rat, while implants in contralateral limbs served as the unloaded controls. Hind limbs were imaged with high-resolution micro-CT (12.5 µm voxel size) at 2, 5, 9, and 12 days post-surgery. Trabecular changes over time were detected by 3D image registration allowing for measurements of bone-formation rate (BFR) and bone-resorption rate (BRR). At day 9, mean %BV/TV for loaded and unloaded limbs were 35.5 ± 10.0% and 37.2 ± 10.0%, respectively, and demonstrated significant increases in bone volume compared to day 2. BRR increased significantly after day 9. No significant differences between bone volumes, BFR, and BRR were detected due to implant loading. Although not reaching significance (p = 0.16), an average 119% increase in pull-out strength was measured in the loaded implants. © 2016 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 35:997-1006, 2017.

FOXA1, GATA3 and PPARÉ£ Cooperate to Drive Luminal Subtype in Bladder Cancer: A Molecular Analysis of Established Human Cell Lines.

Discrete bladder cancer molecular subtypes exhibit differential clinical aggressiveness and therapeutic response, which may have significant implications for identifying novel treatments for this common malignancy. However, research is hindered by the lack of suitable models to study each subtype. To address this limitation, we classified bladder cancer cell lines into molecular subtypes using publically available data in the Cancer Cell Line Encyclopedia (CCLE), guided by genomic characterization of bladder cancer by The Cancer Genome Atlas (TCGA). This identified a panel of bladder cancer cell lines which exhibit genetic alterations and gene expression patterns consistent with luminal and basal molecular subtypes of human disease. A subset of bladder cancer cell lines exhibit in vivo histomorphologic patterns consistent with luminal and basal subtypes, including papillary architecture and squamous differentiation. Using the molecular subtype assignments, and our own RNA-seq analysis, we found overexpression of GATA3 and FOXA1 cooperate with PPARÉ£ activation to drive transdifferentiation of a basal bladder cancer cells to a luminial phenotype. In summary, our analysis identified a set of human cell lines suitable for the study of molecular subtypes in bladder cancer, and furthermore indicates a cooperative regulatory network consisting of GATA3, FOXA1, and PPARÉ£ drive luminal cell fate.

Practicing handoffs early: Applying a clinical framework in the anatomy laboratory.

The anatomy laboratory provides an ideal environment for the integration of clinical contexts as the willed-donor is often regarded as a student's "first patient." This study evaluated an innovative approach to peer teaching in the anatomy laboratory using a clinical handoff context. The authors introduced the "Situation, Background, Assessment, Recommendation" (SBAR) handoff framework within the anatomy laboratory. Study participants included 147 second-year medical students completing the head and neck portion of an anatomy course. The authors used mixed methods to evaluate the impact of the anatomic SBAR on the student anatomy laboratory experience. Qualitative analysis of student evaluations revealed three themes which emerged from students' summaries of their anatomic handoff experiences: Learning-by-teaching; Acquiescing to doing more with less; and Distrust of the peer handoff process. All the themes demonstrated that the anatomic handoff encouraged students' focus on the knowledge preparation and reflection. Closed question analysis suggested that that students' perceptions of handoff usefulness were tied to deeper learning strategies. The handoff provided a mechanism for promoting students' focus on anatomical relationships and facilitated students' learning of transferable clinical skills. Together, these results suggest that the introduction of a handoff process in anatomy education provided both a mechanism for learning anatomy and a unique opportunity for early exposure to an essential clinical skill. This clinical and basic science integration may serve as a vertical integration thread which can be woven throughout undergraduate medical education. Future study will focus on exploring the long-term impacts and learning outcomes of this integration. Anat Sci Educ 9: 476-487. © 2016 American Association of Anatomists.

Raising awareness of the hidden curriculum in veterinary medical education: a review and call for research.

The hidden curriculum is characterized by information that is tacitly conveyed to and among students about the cultural and moral environment in which they find themselves. Although the hidden curriculum is often defined as a distinct entity, tacit information is conveyed to students throughout all aspects of formal and informal curricula. This unconsciously communicated knowledge has been identified across a wide spectrum of educational environments and is known to have lasting and powerful impacts, both positive and negative. Recently, medical education research on the hidden curriculum of becoming a doctor has come to the forefront as institutions struggle with inconsistencies between formal and hidden curricula that hinder the practice of patient-centered medicine. Similarly, the complex ethical questions that arise during the practice and teaching of veterinary medicine have the potential to cause disagreement between what the institution sets out to teach and what is actually learned. However, the hidden curriculum remains largely unexplored for this field. Because the hidden curriculum is retained effectively by students, elucidating its underlying messages can be a key component of program refinement. A review of recent literature about the hidden curriculum in a variety of fields, including medical education, will be used to explore potential hidden curricula in veterinary medicine and draw attention to the need for further investigation.

Teaching laboratory rodent research techniques under the tenets of situated learning improves student confidence and promotes collaboration.

A targeted needs assessment at our institution revealed that the online system used to train researchers on performing techniques with animals did not provide opportunities to practice skills, introduce learners to animal care staff, nor satisfactorily support researchers' needs to become comfortable with laboratory animal species. To correct these deficiencies, a series of hands-on training sessions, framed theoretically in situated learning, was developed. This theoretical framework asserts that learning for everyday living (in this case, performing laboratory animal techniques) happens when people interact within the community while using the 'tools at hand' (that is, the instruments and jargon of the field). From this perspective, the students work alongside the instructor as apprentices. The instructor creates increasingly challenging learning opportunities as students work toward independently performing techniques. To test our hypothesis that teaching from this perspective improves comfort levels with laboratory animals and promotes collaborative relationships between animal care and research personnel, a mixed-method design involving online surveys (first survey, n = 45; second survey, n = 35) and semistructured interviews (n = 10) was used. Quantitative results revealed that students became more comfortable with laboratory animals and were more likely to contact animal care personnel due to participating in the training program. The qualitative arm of the study identified specific features of the training program that improved comfort levels for students (seeing then doing, working in small groups, learning within a comfortable environment, and building collegial relationships). These results support teaching rodent research techniques from the practical and theoretical approach of situated learning.

Atipamezole reverses ketamine-dexmedetomidine anesthesia without altering the antinociceptive effects of butorphanol and buprenorphine in female C57BL/6J mice.

Butorphanol and buprenorphine are common analgesics used in laboratory mice. Inadvertent attenuation of the antinociceptive effects of these analgesics via the administration of an anesthetic reversal agent could result in postprocedural pain and distress, with subsequent negative effects on animal welfare, study outcomes, and regulatory compliance. This study was undertaken to determine whether atipamezole reverses ketamine-dexmedetomidine anesthesia and alters the antinociceptive effects of butorphanol and buprenorphine in female C57BL/6J mice. Atipamezole reliably reversed the anesthetic effects of ketamine-dexmedetomidine, and mice were ambulatory 17.4 ± 30.6 min after administration of the α2-adrenoreceptor antagonist. Atipamezole alone had no significant effect on tail-flick latency and did not alter the antinociceptive properties of butorphanol or low-dose (0.05 mg/kg) or high-dose (0.1 mg/kg) buprenorphine in female C57BL/6J mice. After reversal of ketamine-dexmedetomidine anesthesia, tail-flick latency at 30, 60, and 150 min after analgesic treatment differed significantly between mice treated with atipamezole alone and those given atipamezole followed by butorphanol or high-dose buprenorphine. These results suggest that the analgesic effects of butorphanol and buprenorphine are not affected by atipamezole. Buprenorphine (0.1 mg/kg) administered 30 min prior to or at the time of anesthesia resulted in a greater magnitude of antinociception after antagonism of anesthesia than when given at the time of reversal. Given these results, we recommend the use of ketamine-dexmedetomidine anesthesia with buprenorphine administered either preemptively or at the time of anesthetic induction to provide a defined period of surgical anesthesia that is effectively reversed by atipamezole.