bluuebunny/crossref_metadata_2025 huggingface dataset:
The Current Status and Problems with the Implementation of Interprofessional Education in Japan.
Authors: Ogawa Sachiko , Takahashi Yoshinori , Miyazaki Misako
Published: March 2018

Background: Although interprofessional education (IPE) has come to be considered essential in health and social care education programs, most IPE programs in Japan focus on clinical settings. However, following the 2011 Great East Japan Earthquake, IPE programs are considered essential for community development, especially in disaster-affected areas. To identify key issues for the development of IPE, we aimed to clarify the current status of IPE programs and problems in their implementation using an original questionnaire. Methods and Findings: The targets were 865 undergraduate courses that qualify students to take national registered health/social care examinations. Effective responses were received from 284 targets. Of these 284 respondents, 103 respondents had already implemented an IPE program and 181 respondents had not. Among the 103 respondents who had already implemented an IPE program, we found a tendency to collaborate with partners in clinical settings or in social settings. Furthermore, respondents who had implemented or were planning to implement an IPE program had difficulty with ‘interdisciplinary and/or extramural collaboration’ and ‘educational factors’. Conclusions: These difficulties could be considered barriers to developing effective IPE programs for community-based collaboration between health and social care professionals. Future research should investigate more specific solutions to these problems.
The Effects of Intraprofessional Collaborative Case Based Learning: A Cohort Study of Student Physical Therapist and Physical Therapist Assistant Perceptions of the Physical Therapist Role
Authors: Colgrove Yvonne Marie , Walton Lori Maria , VanHoose Lisa Delores
Published: March 2018

Background: The changing nature and complex regulation of healthcare require the efficient use of resources, including the appropriate delegation and supervision of the physical therapist assistant (PTA). Knowledge of the scope of PTA practice introduced in the academic curriculum is mandated for entry-level practice. This study assessed the effect of a collaborative case-based educational intervention within the didactic curriculums of a physical therapy (PT) and PTA program on student knowledge of PTA scope of practice.Methods and Findings: A pre- and post-test research design was used. Students completed a validated survey exploring their perceptions of the PTA role before beginning the case study. The case study was a classroom assignment followed by instructional prompts requiring interactions between student cohorts three times over four weeks. Following case study completion, students completed the same survey. Independent and paired samples t-tests detected significant differences between and within groups (p < .05).Conclusions: Based on the results, the case-based instructional model was efficacious in teaching both student cohorts about the role of the PTA. The impact was greater on the accuracy of the PT students, but PTA students became less uncertain in their perceptions. The effect of the clinical learning environment should be investigated to determine the impact on student perception of PTA role delineation following didactic instruction.
Using a Research-Informed Interprofessional Curriculum Framework to Guide Reflection and Future Planning of Interprofessional Education in a Multi-Site Context
Authors: Moran Monica Catherine , Steketee Carole , Forman Dawn , Dunston Roger
Published: March 2018

Background: Over the past two years health educators in Australia have benefited from funding made available from national organizations such as the Office of Learning and Teaching (OLT) and Health Workforce Australia (HWA). Funded research has been conducted into educational activities across the country that aim to promote integrated and sustainable interprofessional learning.Methods and Findings: A collaboration between multiple stakeholders led to theestablishment of a consortium of nine universities and interprofessional organizations. This collaboration resulted in a series of research studies and the development of a conceptual framework to guide the planning and review of interprofessional health curricula. A case study of the development of a suite of health education programs at a regional university in Australia is used to demonstrate how the framework can be used to guide curricular reflection and to plan for the future. Shedding a light on interprofessional health education activities across multiple sites provides a rich picture of current practices and future trends. Commonalities, gaps, and challenges become much more obvious and allow for the development of shared opportunities and solutions.Conclusions: The production of a shared conceptual framework to facilitate interprofessional curriculum development provides valuable strategies for curricular reflection, review, and forward planning.
Shadowing: Interprofessional Learning
Authors: Vasset Frøydis Perny , Almås Synnøve Hofseth
Published: March 2018

Background: The Norwegian government has indicated that health and socialstudies should emphasize interprofessional collaborative learning (IPL), especiallyin clinical placements. Through IPL, students have the opportunity to gain insightinto other professional responsibilities and minimize negative stereotypes. Thismight improve collaboration across professional boundaries. Professionals withcollaborative competence might solve complex health problems, and thus improvethe quality of healthcare. The objectives of this article are to investigate the IPLexperiences nursing students acquire through shadowing practice with differentprofessionals in home care.Methods and Findings: To develop a model for IPL, 12 nursing students spent five days shadowing four different healthcare professionals working in home care. At the end of the pedagogical intervention, the students reflected on the practice and the role of the different professionals they had followed. To investigate how the students experienced interprofessional shadowing practice, the reflective notes were analyzed, templates for the selected professionals were drawn up, and four focus group interviews were conducted The results showed that students has acquired knowledge of other professions’ responsibilities and were aware of thneed for an interprofessional approach to home care.Conclusions: This kind of shadowing might be an ideal model for educationalinstitutions seeking to implement IPL.
Bridging the Location Gap: Physician Perspectives of Physician - Pharmacist Collaboration in Patient Care (BRIDGE Phase II)
Authors: Hager Keri D. , Uden Donald , Tomaszewski Daniel
Published: March 2018

Background: To optimize patient outcomes, the patient-centred medical home model emphasizes comprehensive team-based care. Pharmacists are qualified to enhance appropriate medication use and help improve patient outcomes through provision of medication therapy management (MTM) services. To optimally provide MTM, pharmacists must effectively collaborate with physicians. This study explored factors that influence pharmacist-physician collaboration.Methods and Findings: A convenience sample of five physicians participated in semi-structured interviews and the resulting data were analyzed using qualitative methods. Transcripts of the interviews were independently coded for themes by two researchers. Five themes emerged: trustworthiness, role specification, relationship initiation, effects on practice, and professional awareness/expectations.Conclusions: Overall interviewees spoke positively about pharmacists; however, when discussing collaboration, they spoke almost exclusively about pharmacists within their clinic. Since most pharmacists practice outside of clinics, bridging the location gap is imperative for collaboration. In addition, physicians lacked an overall understanding of pharmacists’ training and clinical capacity. This may inhibit pharmacists from participating to their full professional capability within integrated healthcare teams. One approach to resolve this lack of physician understanding of pharmacists’ role and value may be to co-educate health professional students. Further research is needed to explore ways to improve interprofessional collaborative care.
Changes in Pharmacy Students After Experiencing Interprofessional Education Activities
Authors: Tolleson Shane R. , Ismail Nadia J. , Gill Anne C. , Dello Stritto Rita A. , Hatfield Mark D. , Teal Cayla R. , Nelson Elizabeth A. , Hatfield Catherine L.
Published: March 2018

Background: Interprofessional education (IPE) has become increasingly important among healthcare professionals. The purpose of this study was to assess the impact of IPE experiences on pharmacy students’ perceived level of comfort with and level of reliability regarding other healthcare professionals. Methods and Findings: A survey of third- and fourth-year pharmacy students (N = 190) was analyzed using descriptive statistics and bivariate analyses to address the study objectives. Students experienced a significant increase in their level of comfort with questioning other professions after IPE (pre IPE: 5.45 ± 2.24 and post IPE: 7.22 ± 1.62, p<0.01) and being questioned by other professions (pre IPE: 5.39 ± 2.24 and post IPE: 7.04 ± 1.68, p<0.01). Students also experienced a significant increase in the perceived reliability and accuracy of information from nurses after IPE (pre IPE: 5.91 ± 1.82 and post IPE: 6.41 ± 1.62, p = 0.01). Conclusions: Pharmacy students who experienced any IPE perceived themselves as more comfortable with questioning and being questioned by other healthcare professionals than students without IPE. Additional research is needed to more clearly define the benefits of IPE and how it should be designed.
Examining Professional Stereotypes in an Interprofessional Education Simulation Experience
Authors: Rudd Alison Bealle , Estis Julie M , Pruitt Bill , Wright Theresa
Published: March 2018

Background: Interprofessional education (IPE) provides a platform for early professional socialization, potentially affecting the accuracy of stereotypes among health professions students. The purpose of this study was to implement an interprofessional simulation with nursing, respiratory therapy (RT), and speech language pathology (SLP) students, and using the Student Stereotype Rating Questionnaire, evaluate how an IPE simulation approach may alter stereotypes that learners carry with them related to themselves and professions other than their own.Methods and Findings: Participants rated the extent to which they believe attributes, based on nine professional characteristics, apply to either their own profession (autostereotypes), other professions (heterostereotypes), or their own profession as seen by others (perceived autostereotypes) with the Student Stereotype Rating Questionnaire (SSRQ). A quasi-experimental pretest-posttest design was used, and descriptive and analytical statistics conducted within and across groups. Participant impressions of the IPE experience are presented. Main limitations included smaller sample size of RT and SLP participants.Conclusions: Results showed a significant difference from pre- to post-IPE simulation in nursing heterostereoptype, autostereotype, and perceived autostereotype scores. No significant difference was seen in hetereostereotypes of RT and SLP students. Overall, student impressions were positive. Recommendations include study replication for larger sample size.
Emotional Intelligence and Teamwork Skills Among Undergraduate Engineering and Nursing Students: A Pilot Study
Authors: Lee Charlotte , Bristow Michele , Wong Jason C.
Published: March 2018

Background: Healthcare operations require collaboration between healthcare and non-healthcare professionals.Objectives: To examine: a) the effectiveness of a case-based interprofessional education (IPE) workshop, and b) the association between emotional intelligence (EI) and teamwork skills in nursing and industrial engineering trainees.Methods: Undergraduate nursing and industrial engineering students (N = 42) participated in a workshop that requires the integration of knowledge from both disciplines. Previously validated instruments were used to assess EI, empathic listening and teamwork skills pre- and post-workshop.Findings: While results of the multivariate analysis of variance suggested no difference in study variables before and after the workshop, findings from Pearson’s correlation revealed more pairs of significant correlations post-workshop. Positive association between EI and teamwork skills was found post-workshop.Conclusion: IPE workshops beyond conventional healthcare professions are feasible and welcomed by trainees. EI should be considered an antecedent of teamwork.
Abordagem cirúrgica do prolapso retal em felino: Relato de caso
Authors: Viliotti Tatianne , Lima Anne Nelizia Holanda de , Rodrigues Ingrid Rabelo , Feitosa Aline Silveira , Ferraz Richard Elaino de Oliveira , Santos Rebeca de Melo , Ceni Sarah Ary , Fernandes Maria Ester Crispim Nogueira
Published: March 2018

Rectal prolapse is a protrusion or eversion of the rectal mucosa from the anus and it is more reported in young animals. It is usually a consequence of underlying disorders intestinal diseases that cause diarrhoea and. The present study reports a case of rectal prolapsed in a cat puppy associated to necrose of the rectum, disquesia and emaciation. A feline S.R.D, 2 months old, was attended at the Metropolitan Veterinary Hospital, Caucaia, Ceará, Brazil, presenting the prolapsed rectum, tenesmus, dyskinesia, anorexia, emaciation, prostration and regions of necrosis in the prolapsed rectum. Haematological and biochemical tests revealed anemia and ALT increases. The surgical treatment consisted resection of restum and it was made an anastomose throue in the application of cross anchors and resection of the prolapsed mass and in the application of separate single sutures to make the anastomosis. The patient was reassessed in the following weeks and showed a return to the normal physiological functions of defecation. It is concluded that rectal amputation may be an alternative for the surgical treatment of rectal prolapse in felines.
Diagnóstico e tratamento de lúpus eritematoso discoide canino: Relato de caso
Authors: Da silva Vanessa , Monteiro Cynthia , Silva Michelle , Carneiro Raimundo , Lucena Lorena
Published: March 2018

The Lupus Erythematosus immune complex disease (LE) consists of a set of autoimmune dermatopathies that can manifest in two main clinical presentations: Systemic Lupus Erythematosus (SLE) and Discoid Lupus Erythematosus (DLE). The clinical diagnosis of DLE can be challenging, due to the variety of symptoms that can occur both in lupus and in other dermatological conditions, which interferes with the early diagnosis and consequently a delay in the treatment and improvement of the patient. This study aims to report the importance of histopathological examination in the definitive diagnosis for treatment and favorable prognosis of canine poodle with DLE. A poodle dog was attended at the Metropolitan Veterinary Polyclinic - HVM, presenting lesion in nasal area with previous treatments and no apparent improvment of the lesion. Laboratory and cytological exams were done to exclude other skin diseases. After an inconclusive result in the cytology, the histopathological examination was requested, to define the exact diagnosis and exclusion of other dermatopathies. The histopathological confirmed the diagnosis of Discoid Lupus Erythematosus. Treatment with corticosteroids at the dose of 1.0mg / kg, antioxidant vitamins and sunscreen was instituted. After 10 days, the dose increased to 2.5mg / kg, which is the immunosuppressive dose. The animal presented a response with a decrease in the crustal region of the lesion in the nasal plane.
Relação homem-animal durante procedimento clínico cirúrgico e pós-operatório em bezerro Holandês Preto e Branco acometido por atresia anal: Relato de Caso
Authors: Amaral Jackson Barros do , Trevisan Guilherme
Published: March 2018

This relate tells the objective of describing human-animal relation during clinical-surgical treatment and postoperative treatment in Holstein calf treated by anal atresia and its possible effects on well-being and gaining weight. Anal atresia is a congenital malformation with occlusions in the challenge and anus that occurs in animals and in human. The well-being of animals afflicted by pain and suffering in the postoperative can be influenced by the human-animal relationship. The animal was born in 2016 and after 48 hours, it was found atresia anal grau II, suggestive of pain and suffering. Emergency surgery was performed such as sedation, local anesthesia and epidural. The evaluation of the welfare was performed by clinical, behavioral, and physiological and weight gain hair. In the postoperative antibiotic, anti-inflammatory and polyvitamins were administered. After surgery, elimination of gases and mechanics was observed, following normal evacuations until discharge. The surgical wound showed good conditions; however, the feces presented blood strains, fibrin and rectal tense. The animal normalized the appetite and appeared active and how vigorous at the end of the surgery, explored the environment and used the soft bed at moments of rest and vocalize before breastfeeding. Chopped hay and ration were available, which favored an alteration in the consistency of the feces, going from semipastosa to pasture. In the postoperative, human-animal interaction was established, where it was possible to observe positive accounts such as dropping out, dropping fingers, scratching humans, demonstrating playful behaviors such as persecutions, acrobatics, jump-in, running, manipulations as targets, air and head. The physiological indicators were normalized. The weight gain was 11 kg in 25 days. It is concluded that during the first 30 days, the surgical and operative clinical treatment the animal presented a good clinical evolution and that humanitarian care as "positive" actions in the human-animal relationship influenced the welfare and gain of weight.
Toxicose por picada de abelhas em cão
Authors: Silva Sousa Jéssica
Published: March 2018

Accidents due to bee stings in dogs occur frequently, however, there are few cases described in the literature. This report describes a fatal case of a canine, with signs of systemic toxic reaction due to a bees accident. The main clinical signs were hemolysis and hemoglobinuria. Other findings included tachycardia, and dyspnoea as well as the presence of stings in the animal's body. Despite the institution of clinical treatment, the animal died two hours later.
On distances in generalized Sierpiński graphs
Authors: Estrada-Moreno Alejandro , Rodríguez-Bazan Erick , Rodríguez-Velázquez Juan
Published: March 2018

In this paper we propose formulas for the distance between vertices of a generalized Sierpi?ski graph S(G, t) in terms of the distance between vertices of the base graph G. In particular, we deduce a recursive formula for the distance between an arbitrary vertex and an extreme vertex of S(G, t), and we obtain a recursive formula for the distance between two arbitrary vertices of S(G, t) when the base graph is triangle-free. From these recursive formulas, we provide algorithms to compute the distance between vertices of S(G, t). In addition, we give an explicit formula for the diameter and radius of S(G, t) when the base graph is a tree.
The relevance of the social context for media coverage of victims
Authors: Pavicevic Olivera , Bulatovic Aleksandra
Published: March 2018

Media coverage of human suffering caused by trauma, illness, poverty and
disasters worldwide takes up a significant part of media coverage and
affects the identity of all actors, both active and passive. Media
presentation shapes our thinking and reasoning at the group level, and has
far reached impact. The social context of media reporting on victims is
shaped by the capacity of the social system to recognize, acknowledge,
strengthen and protect the victim. As an aspect of this capacity, media
coverage of victims is indicator of victimization visibility. This paper
discusses nature of media reporting as a phenomenon appearing in various
forms as per given sources of victimization, and the ethical aspects of
media presentation of victims. The aim of the paper is to contribute to the
critical analysis of the media coverage of victims, by reconsidering an
increased visibility of victims and their suffering, which is generated by
media reporting, and whose dominant characteristic is presenting victims in
the interpretative and performing manner.
Normative framework of media reporting on victims of crime in the Republic of Serbia
Authors: Jemuovic Jelena
Published: March 2018

The topic of this paper is the normative framework of media reporting on
victims of crime in the Republic of Serbia. The aim of the paper is to
provide an overview of international and national legal instruments that
regulate the way the media should report on this important issue and to
assess the level of compliance of the Serbian legal framework with
international standards in this field. The freedom of media is inviolable in
every democratic society, but there are clear rules that must be respected.
Essential are the ethical standards that journalists have to comply with in
their work, which are also analyzed in the paper. After considering relevant
regulations governing media reporting on victims of crime, it is concluded
that the national legal framework on media reporting on crime victims in
Serbia is in compliance with relevant international instruments.
Femicide in partnership relations
Authors: Spasic Danijela , Kolarevic Dag , Lukovic Zoran
Published: March 2018

In numerous studies in the world, the existence of femicide as a specific
form of homicide has been confirmed, indicating its conditions and
consequences and the strongest risk factors. The subject of this paper is
femicide in intimate partnership, and the aim is to point to the specific
characteristics of femicide in intimate parner relationships in Serbia, i.e.
to present and analyse the data obtained in the empirical research of this
form of murder. The empirical research of femicide in intimate partner
relationships in Serbia was conducted in 2016 on the sample of 153 cases of
murder of women in the context of intimate partnership, which occurred on the
territory of Serbia (without Kosovo and Metohija) in the period from 2001 to
2015. The objectives of the research were to determine the existence and
identification of risk factors for femicide. The data was collected from the
criminal reports filed by the Ministry of Interior of the Republic of Serbia.
The findings of the study confirmed the results of other studies in regard
prevalence of femicide and the existence of the following risk factors:
exposure of women to chronic intimate partner violence, the availability of
firearms, the influence of alcohol and psychoactive substances abuse, the
presence of psychological disorders and illness and suicidal tendencies of
perpetrators. By applying multidimensional scaling in analyzing elements of
crime offending the specific position of suicides that occur in certain
number of cases after femicide was pointed out. There was a slight tendency
of absence of history of violence in cases where perpetrator of femicide
committed suicide.
Near-Infrared Optical Imaging of Ovarian Cancer Xenografts with Novel α3-Integrin Binding Peptide “OA02”
Authors: Aina Olulanu H. , Marik Jan , Gandour-Edwards Regina , Lam Kit S.
Published: March 2018

Through screening of random one-bead one-compound (OBOC) libraries, we previously identified cyclic peptides with the cDGXGXXc motif that bind to α3 integrin subunit on ovarian adenocarcinoma cell lines ES-2, SKOV-3, and CaOV-3. We subsequently synthesized two secondary libraries based on this motif and identified new peptides that bound with a higher affinity to these cell lines. One of the peptides identified from the 20% “down-substituted” focused library was the cdG-HCit-GPQc (“OA02”) peptide. The goal of this study was to determine whether this peptide labeled with near-infrared probes could be detected after intravenous injection in ovarian tumor-bearing mice and if it would selectively localize in the tumor. Three different forms of this peptide were synthesized, “OA02”-biotin (noncovalently linked to streptavidin-Cy5.5); “OA02”-Cy5.5 and “OA02”-AlexaFluo 680. Using a KODAK IS2000MM image station, these peptide probes were used at the near-infrared (NIR) spectra to image nude mice bearing ES-2 (α3 integrin positive) and Raji (α3 integrin negative) xenografts. The peptide probe displayed highly specific tumor uptake within 15 min, which lasted for 70 min for “OA02”-Cy5.5 and “OA02”-AlexaFluo 680 and for 24 hours for “OA02”-biotin-streptavidin-Cy5.5. Some kidney and bladder signal were noted. Prior injection with anti-α3 monoclonal antibody blocked the binding of this peptide to the ES-2 tumors.
Cellular Imaging of Inflammation in Atherosclerosis Using Magnetofluorescent Nanomaterials
Authors: Jaffer Farouc A. , Nahrendorf Matthias , Sosnovik David , Kelly Kimberly A. , Aikawa Elena , Weissleder Ralph
Published: March 2018

Objective: Magnetofluorescent nanoparticles (MFNPs) offer the ability to image cellular inflammation in vivo. To better understand their cellular targeting and imaging capabilities in atherosclerosis, we investigated prototypical dextran-coated near-infrared fluorescent MFNPs in the apolipoprotein E-deficient (apo E−/−) mouse model.

Methods and Results: In vitro MFNP uptake was highest in activated murine macrophages ( p < .001). Apo E−/− mice ( n = 11) were next injected with the MFNP (15 mg/kg iron) or saline. In vivo magnetic resonance imaging (MRI) demonstrated strong plaque enhancement by the MFNPs ( p < .001 vs. saline), which was confirmed by multimodality ex vivo MRI and fluorescence reflectance imaging. On fluorescence microscopy, MFNPs were found in cellular-rich areas of atheroma and colocalized with immunofluorescent macrophages over endothelial cells and smooth muscle cells ( p < .001).

Conclusions: Here we show that (1) the in vitro and in vivo cellular distribution of atherosclerosis-targeted MFNPs can be quantified by using fluorescence imaging methods; (2) in atherosclerosis, dextranated MFNPs preferentially target macrophages; and (3) MFNP deposition in murine atheroma can be noninvasively detected by in vivo MRI. This study thus provides a foundation for using MFNPs to image genetic and/or pharmacological perturbations of cellular inflammation in experimental atherosclerosis and for the future development of novel targeted nanomaterials for atherosclerosis.
Feasibility and Limits of Magnetically Labeling Primary Cultured Rat T Cells with Ferumoxides Coupled with Commonly Used Transfection Agents
Authors: Berger Cedric , Rausch Martin , Schmidt Philipp , Rudin Markus
Published: March 2018

Visualization and quantification of inflammatory processes is of high importance for early diagnosis of a multitude of diseases. Magnetic resonance imaging (MRI) using iron oxide (FeO) nanoparticles as contrast agents allows the study of macrophage infiltration during inflammation in a variety of tissues. Macrophages are effectors of the immune response, their appearance being orchestrated by activated T lymphocytes. Therefore, tracking of labeled T lymphocytes, which initiate the immune process, should enable earlier detection of tissue inflammation. In this study, we investigate the feasibility of specifically labeling harvested T cells by using dextran-coated FeO nanoparticles and commonly available transfection agents (TAs). Physicochemical properties of the newly formed FeO/TA vesicles were determined as well as their cell toxicity and their T cell activation potential. The labeling efficiency of each FeO/TA combination was evaluated by measuring the transverse MRI relaxation rate R2 by X-ray spectroscopy and magnetic selection. Toxicity and labeling efficacy differed significantly among TAs. The best results were achieved by using polyamine TAs and in particular by using poly-l-lysine at a concentration of 1.5 µg/mL administered in combination with 22.5 µg iron/mL. By using this protocol, up to 60% of harvested T cells could be labeled. Microscopic investigation revealed FeO/TA nanoparticles not only localized within the cytoplasma of the cells but also sticking to the outer membrane surface.
Light Emission Requires Exposure to the Atmosphere in Ex Vivo Bioluminescence Imaging
Authors: Inoue Yusuke , Izawa Kiyoko , Tojo Arinobu , Sekine Rieko , Okubo Toshiyuki , Ohtomo Kuni
Published: March 2018

The identification of organs bearing luciferase activity by in vivo bioluminescence imaging (BLI) is often difficult, and ex vivo imaging of excised organs plays a complementary role. This study investigated the importance of exposure to the atmosphere in ex vivo BLI. Mice were inoculated with murine pro-B cell line Ba/F3 transduced with firefly luciferase and p190 BCR-ABL. They were killed following in vivo BLI, and whole-body imaging was done after death and then after intraperitoneal air injection. In addition, the right knee was exposed and imaged before and after the adjacent bones were cut. Extensive light signals were seen on in vivo imaging. The luminescence disappeared after the animal was killed, and air injection restored the light emission from the abdomen only, suggesting a critical role of atmospheric oxygen in luminescence after death. Although no substantial light signal at the right knee was seen before bone cutting, light emission was evident after cutting. In conclusion, in ex vivo BLI, light emission requires exposure to the atmosphere. Bone destruction is required to demonstrate luciferase activity in the bone marrow after death.
Chloroquine Treatment Increases Detection of 5-Fluorouracil-Induced Apoptosis In Vivo
Authors: Nyati Mukesh K. , Feng Felix Y. , Kanade Vinayak D. , Nayak Rabindranath
Published: March 2018

Although apoptosis can be readily assessed in vitro with a variety of techniques, the detection of apoptosis in the in vivo setting poses a much more difficult proposition. Apoptosis in an organism is followed almost inevitably by rapid clearance of dying cells via phagocytosis, thus limiting the ability to analyze apoptosis in vivo using classical techniques. To address this issue, we developed a method to enhance in vivo apoptosis detection using pretreatment with chloroquine, an inhibitor of macrophage activity, in Swiss albino mice. This technique resulted in a significant increase in the accumulation of apoptotic cells induced by 5-fluorouracil, as detected by propidium iodide staining in solid and ascitic forms of Ehrlich ascitic tumors and in bone marrow cells. We further validated our technique using DNA fragmentation and endonuclease assays. Our results demonstrated that chloroquine pretreatment can significantly enhance accumulation of apoptotic cells in organisms, and we envision combining this method with modern imaging techniques to optimize in vivo detection of apoptosis.
Choline Transporter as a Novel Target for Molecular Imaging of Cancer
Authors: Hara Toshihiko , Bansal Aditya , DeGrado Timothy R.
Published: March 2018

Abnormalities of choline processing in cancer cells have been used as a basis for imaging of cancer with positron emission tomography and magnetic resonance spectroscopy. In this study, the transport mechanism for choline was investigated in cultured PC-3 prostate cancer cells. Furthermore, tritiated hemicholinium 3 (HC-3), a well-known inhibitor of choline transport, was studied as a prototypic molecular imaging probe in PC-3 cells and 9L glioma–bearing rats. [3H]Choline uptake by PC-3 cells was found to have both facilitative and nonfacilitative components. Facilitative transport was characterized by partial sodium dependence and intermediate affinity ( KM = 9.7 ± 0.8 μM). HC-3 inhibited choline with a KI of 10.5± 2.2 μM. Ouabain (1 mM) caused a 94% reduction in choline uptake. At physiologic choline concentration, phosphocholine was the rapid and predominant metabolic fate. The binding of [3H]HC-3 to PC-3 cells was rapid and specific (competitively blocked with unlabeled HC-3). Biodistribution of [3H]HC-3 in 9L glioma–bearing rats showed the ranking of uptake to be kidney > lung > tumor > liver > skeletal muscle ≈ blood > brain. In comparison with [14C]choline, [3H]HC-3 showed over twofold higher tumor uptake and favorable uptake ratios of tumor to blood, tumor to muscle, tumor to lung, and tumor to liver. The data demonstrate the quantitative importance of an intermediate-affinity, partially sodium-dependent choline transport system on choline processing in PC-3 cancer cells. The biodistribution properties of [3H]HC-3 in tumor-bearing rats encourage the development of molecular imaging probes based on choline transporter binding ligands.
Cardiac Micro–Computed Tomography for Morphological and Functional Phenotyping of Muscle LIM Protein Null Mice
Authors: Badea Cristian T. , Hedlund Laurence W. , Mackel Julie F. Boslego , Mao Lan , Rockman Howard A. , Johnson G. Allan
Published: March 2018

The purpose of this study was to investigate the use of micro–computed tomography (micro-CT) for morphological and functional phenotyping of muscle LIM protein (MLP) null mice and to compare micro-CT with M-mode echocardiography. MLP null mice and controls were imaged using both micro-CT and M-mode echocardiography. For micro-CT, we used a custom-built scanner. Following a single intravenous injection of a blood pool contrast agent (Fenestra VC, ART Advanced Research Technologies, Saint-Laurent, QC) and using a cardiorespiratory gating, we acquired eight phases of the cardiac cycle (every 15 ms) and reconstructed three-dimensional data sets with 94-micron isotropic resolution. Wall thickness and volumetric measurements of the left ventricle were performed, and cardiac function was estimated. Micro-CT and M-mode echocardiography showed both morphological and functional aspects that separate MLP null mice from controls. End-diastolic and -systolic volumes were increased significantly three- and fivefold, respectively, in the MLP null mice versus controls. Ejection fraction was reduced by an average of 32% in MLP null mice. The data analysis shows that two imaging modalities provided different results partly owing to the difference in anesthesia regimens. Other sources of errors for micro-CT are also analyzed. Micro-CT can provide the four-dimensional data (three-dimensional isotropic volumes over time) required for morphological and functional phenotyping in mice.
Image-Guided Analyses Reveal that Non-CD4 Splenocytes Contribute to CD4+ T Cell–Mediated Inflammation Leading to Islet Destruction by Altering Their Local Function and Not Systemic Trafficking Patterns
Authors: Lee Mi-Heon , Lee Wen-Hui , Van Yanghao , Contag Christopher H. , Liu Chih-Pin
Published: March 2018

Recruitment of CD4+ T cells into islets is a critical component of islet inflammation (insulitis) leading to type 1 diabetes; therefore, determining if conditions used to treat diabetes change their trafficking patterns is relevant to the outcome. Cotransfer of CD4+BDC2.5 (BDC) cells with non-CD4 splenocytes obtained from newly diabetic NOD mice, but not when they are transferred alone, induces accelerated diabetes. It is unclear whether these splenocytes affect diabetes development by altering the systemic and/or local trafficking and proliferation patterns of BDC cells in target and nontarget tissues. To address these questions, we developed an animal model to visualize BDC cell trafficking and proliferation using whole-body in vivo bioluminescence imaging and used the images to direct tissue sampling for further analyses of the cell distribution within tissues. The whole-body, or macroscopic, trafficking patterns were not dramatically altered in both groups of recipient mice. However, the local patterns of cell distribution were distinct, which led to invasive insulitis only in cotransferred mice with an increased number of islet-infiltrating CD11b+ and CD11c+ cells. Taken together, the non-CD4 splenocytes act locally by promoting invasive insulitis without altering the systemic trafficking patterns or proliferation of BDC cells and thus contributing to diabetes by altering the localization within the tissue.
HSP70-Inducible hNIS-IRES-eGFP Reporter Imaging: Response to Heat Shock
Authors: Che Jiantu , Doubrovin Mikhail , Serganova Inna , Ageyeva Lyudmila , Beresten Tatiana , Finn Ronald , Blasberg Ronald
Published: March 2018

A retroviral vector pQHSP70/hNIS-IRES-eGFP (pQHNIG70) was constructed containing the hNIS-IRES-eGFP dual-reporter genes under the control of an inducible human heat shock protein (HSP)70 promoter and RG2-pQHSP70/hNIS-IRES-eGFP (RG2-pQHNIG70) transduced cells were generated. Heat-induced expression of both reporter genes in RG2-pQHNIG70 cells was validated by enhanced green fluorescent protein (eGFP) fluorescence-activated cell sorter, in vitro radiotracer assays, and immunoblot and immunocytochemistry. A 2.2- to 6.1-fold (131I−), a 6.1- to 14.4-fold (99mTcO4−), and a 5.1- to 39-fold (fluorescence) increase above baseline was observed in response to graded hyperthermia (39–43°C). Increases in eGFP fluorescence and radiotracer uptake were first noted at 6 hours, reached a maximum at 24 hours, and fell toward baseline at 72 hours. A stable ratio of radiotracer uptake to eGFP fluorescence and to heat shock protein (HSP)70 protein was demonstrated over a wide range of expression levels, induced by different levels of heating. We also demonstrate that the local application of heat on RG2-pQHNIG70 xenografts can effectively induce hNIS and eGFP gene expression in vivo and that this expression can be efficiently visualized by fluorescence, scintigraphic, and micro–positron emission tomography imaging. Endogenous HSP70 protein and reporter expression was confirmed by postmortem tissue evaluations (immunoblot and immunohistochemistry). The pQHNIG70 reporter system can be used to study stress and drug responses in transduced cells and tissues.
Targeting Phosphatidylserine on Apoptotic Cells with Phages and Peptides Selected from a Bacteriophage Display Library
Authors: Shao Ruping , Xiong Chiyi , Wen Xiaoxia , Gelovani Juri G. , Li Chun
Published: March 2018

Phosphatidylserine (PS) is a well-characterized biomarker for apoptosis. Ligands that bind to PS can be used for noninvasive imaging of therapy-induced cell death, particularly apoptosis. In this study, we screened a random 12-mer peptide phage library on liposomes prepared from PS. One clone displaying the peptide SVSVGMKPSPRP (designated as PS3-10) bound to PS approximately 4-fold better than its binding to phosphatidylcholine and 18-fold better than to bovine serum albumin in a solid-phase binding assay. In addition, the binding of the corresponding PS3-10 peptide to PS was significantly higher than that of a scrambled peptide. PS3-10 phages, but not a control 4-2-2 phage, bound to aged red blood cells that had PS exposed on their surface. Binding of PS3-10 phages and PS3-10 peptide to TRAIL-induced apoptotic DLD1 cells was 3.2 and 5.4 times higher than their binding to untreated viable cells, respectively. Significantly, immunohistochemical staining confirmed selective binding of PS3-10 phages to apoptotic cells. Our data suggest that panning of phage display libraries may allow the selection of suitable peptide ligands for apoptotic cells and that PS3-10 peptide may serve as a template for further development of molecular probes for in vitro and in vivo imaging of apoptosis.
Signaling Can Be Uncoupled from Imaging of the Somatostatin Receptor Type 2
Authors: Han Lin , Yang Dan , Kundra Vikas
Published: March 2018

Endogenous and exogenous somatostatin receptors are commonly targeted for imaging using radiopharmaceutical analogues of somatostatin. Ligand binding activates receptor-mediated signaling. We assessed whether somatostatin receptor type 2A (SSTR2A) imaging can be uncoupled from signal transduction. In both human fibrosarcoma (HT1080) and human embryonic kidney (HEK293) cells, reverse transcriptase–polymerase chain reaction and enzyme-linked immunosorbent assay found similar levels of expression of hemagglutinin A tagged SSTR2A (HA-SSTR2A) or the same fusion protein with a deletion of the C-terminus beyond amino acid 314 (HA-SSTR2Δ314). Scatchard analysis demonstrated similar degrees of ligand binding by the wild-type or mutant receptor to 111In-octreotide in both cell pairs. Cyclic guanosine monophosphate (cGMP) production and inhibition of forskolin-induced cylic adenosine monophosphate (cAMP) production were evaluated at the signaling level, and growth inhibition was evaluated at the cellular level before and after stimulation. Unlike wild-type receptor, HA-SSTR2Δ314 was deficient in inhibiting forskolin-induced cAMP production ( p < .05) and in inciting cGMP ( p < .05) production; furthermore, at the cellular level, HA-SSTR2Δ314 was deficient in inhibiting cell proliferation ( p < .05). Yet tumors expressing HA-SSTR2Δ314 could be imaged in vivo. Thus, in vivo imaging of SSTR2 can be uncoupled from cAMP and cGMP signaling as well as growth inhibition.
Detection of Dose Response in Chronic Doxorubicin-Mediated Cell Death with Cardiac Technetium 99m Annexin V Single-Photon Emission Computed Tomography
Authors: Gabrielson Kathleen L. , Mok Greta S.P. , Nimmagadda Srihdar , Bedja Djahida , Pin Scott , Tsao Allison , Wang Yuchuan , Sooryakumar Dhrtti , Yu S. Jianhua , Pomper Martin G. , Tsui Benjamin M.W.
Published: March 2018

The aim of this study was to evaluate whether technetium 99m hydrazinonicotinamide (99mTc-HYNIC)-annexin V single-photon emission computed tomography (SPECT) would detect dose-dependent doxorubicin (DOX)-mediated cell death in the heart compared with functional echocardiography. Adult female Sprague-Dawley rats were treated with DOX (cumulative dose of 15 or 7.5 mg/kg) or saline ( n = 7) and monitored by echocardiography. Rats were injected with 7 to 8 mCi 99mTc-HYNIC-annexin V and imaged 1 hour postinjection using a small animal dual-head SPECT/computed tomography (CT) system with multipinhole technology. Two regions of interest were drawn in the myocardium and soft tissue regions to calculate the cardiac uptake ratio (CUR) of reconstructed images. Myocardium and blood were harvested for radioactivity measurements or TUNEL assay. Biodistribution of 99mTc-HYNIC-annexin V uptake, CUR from SPECT/CT fused cardiac images, and TUNEL of myocardium demonstrated a dose-dependent toxicity response, with the cumulative 15 mg/kg DOX treatment showing the greatest degree of cell death. In contrast, echocardiography detected functional deficits only at the highest DOX dose. In vivo molecular imaging of DOX-induced cardiac toxicity with 99mTc-HYNIC-annexin V detects dose-dependent cell death before ventricular deficits are observed with echocardiography. 99mTc-HYNIC-annexin V SPECT-based molecular imaging may provide an attractive new technique for assessing early changes in myocardial function in patients undergoing DOX therapy.
Imaging Changes in Lymphoid Organs In Vivo after Brain Ischemia with Three-Dimensional Fluorescence Molecular Tomography in Transgenic Mice Expressing Green Fluorescent Protein in T Lymphocytes
Authors: Martin Abraham , Aguirre Juan , Sarasa-Renedo Ana , Tsoukatou Debbie , Garofalakis Anikitos , Meyer Heiko , Mamalaki Clio , Ripoll Jorge , Planas Anna M.
Published: March 2018

Stroke induces a strong inflammatory reaction in the brain and depresses the immune system. We sought to assess longitudinal changes in T-cell numbers in the lymphoid organs of living mice after brain ischemia. Middle cerebral artery occlusion was carried out in transgenic mice expressing green fluorescent protein (GFP+) in the T-cell population under the control of the hCD2 locus control region. Imaging was performed by three-dimensional fluorescence molecular tomography (FMT) before and at several time points after ischemia or sham operation and in controls. At day 7, GFP+ cell content in lymphoid organs was measured postmortem by flow cytometry. GFP+ cell numbers and in vivo FMT signal intensity were reduced at day 7 after ischemia and, to a lesser extent, after sham operation. Linear regression analysis demonstrated that postmortem GFP+ cell numbers and corresponding in vivo FMT data were significantly correlated in the thymus ( r2 = .65, p < .0001) and lymph nodes ( r2 = .67, p < .0001). These relationships allowed inferring the number of GFP+ T cells from in vivo FMT data. The results show the time course reduction of T-cell content in the lymphoid organs of living mice, providing in vivo evidence of lymphoid organ atrophy after stroke and, to a lesser extent, after head surgery with craniectomy and dura mater opening in sham-operated mice.
Choline Autoradiography of Human Prostate Cancer Xenograft: Effect of Castration
Authors: Jadvar Hossein , Gurbuz Alparslan , Li Xiankui , Shahinian Antranik , Conti Peter S.
Published: March 2018

The purpose of this study was to investigate the effects of castration and tracer uptake time interval on the level of radiolabeled choline accumulation in murine-implanted human prostate tumor xenografts using quantitative autoradiography. We implanted androgen-dependent (CWR22) and androgen-independent (PC3) human prostate cancer cells in castrated ( n = 9) and noncastrated ( n = 9) athymic male mice and allowed tumors to grow to 1 cm3. The mice were euthanized at 5, 10, and 20 minutes after injection of 5 µCi [14C]-choline. Mice were prepared for quantitative autoradiography with density light units of viable tumor sections converted to units of radioactivity (nCi/mm2) using calibration. Two-group comparisons were performed using a two-tailed Student t-test with unequal variance and with a significance probability level of less than .05. Two-group comparisons between the means of the tracer uptake level for each tumor type at each of three time points for each of two host types showed that (1) the level of tracer localization in the two tumor types was affected little in relation to the host type and (2) PC3 tumor uptake level tended to increase slowly with time only in the noncastrated host, whereas this was not observed in the castrated host or with CWR22 tumor in either host type. The uptake time interval and castration do not appear to significantly affect the level of radiolabeled choline uptake by the human prostate cancer xenograft.
Stably Integrated luxCDABE for Assessment of Salmonella Invasion Kinetics
Authors: Flentie Kelly N. , Qi Min , Gammon Seth T. , Razia Yasmin , Lui Felix , Marpegan Luciano , Manglik Aashish , Piwnica-Worms David , McKinney Jeffrey S.
Published: March 2018

Salmonella Typhimurium is a common cause of gastroenteritis in humans and also localizes to neoplastic tumors in animals. Invasion of specific eukaryotic cells is a key mechanism of Salmonella interactions with host tissues. Early stages of gastrointestinal cell invasion are mediated by a Salmonella type III secretion system, powered by the adenosine triphosphatase invC. The aim of this work was to characterize the invC dependence of invasion kinetics into disparate eukaryotic cells traditionally used as models of gut epithelium or neoplasms. Thus, a nondestructive real-time assay was developed to report eukaryotic cell invasion kinetics using lux+ Salmonella that contain chromosomally integrated luxCDABE genes. Bioluminescence-based invasion assays using lux+ Salmonella exhibited inoculum dose-response correlation, distinguished invasion-competent from invasion-incompetent Salmonella, and discriminated relative Salmonella invasiveness in accordance with environmental conditions that induce invasion gene expression. In standard gentamicin protection assays, bioluminescence from lux+ Salmonella correlated with recovery of colony-forming units of internalized bacteria and could be visualized by bioluminescence microscopy. Furthermore, this assay distinguished invasion-competent from invasion-incompetent bacteria independent of gentamicin treatment in real time. Bioluminescence reported Salmonella invasion of disparate eukaryotic cell lines, including neoplastic melanoma, colon adenocarcinoma, and glioma cell lines used in animal models of malignancy. In each case, Salmonella invasion of eukaryotic cells was invC dependent.