Targeting NEDD8-activating enzyme is a new approach to treat canine diffuse large B-cell lymphoma.

Canine diffuse large B-cell lymphoma (DLBCL), the most common hematologic malignancy of dogs, is associated with poor overall survival. The lack of conventional chemotherapies with sustainable efficacy warrants investigation of novel therapies. Pevonedistat (MLN4924) is a potent and selective small molecule NEDD8-activating enzyme inhibitor. In human activated B-cell-like (ABC) diffuse large B-cell lymphoma, pevonedistat induces lymphoma cell apoptosis, DNA damage and G1 cell cycle arrest by inhibiting the nuclear factor-κB (NF-κB) pathway. Genomic and transcriptomic studies showed that the NF-κB pathway is deregulated in canine DLBCL. Our results showed that pevonedistat treatment significantly reduces the viability of canine DLBCL cells by inducing G1 cell cycle arrest and apoptosis. Pevonedistat treatment inhibits NF-κB pathway activation and downregulates NF-κB target genes in canine DLBCL. Moreover, administration of pevonedistat to mice bearing canine DLBCL xenograft tumours resulted in tumour regression. Our in vivo and in vitro studies provide justification for future clinical application of pevonedistat as a potential new anti-cancer therapy that may benefit both canine and human species.

Prognostic awareness and communication preferences among caregivers of patients with malignant glioma.

OBJECTIVE: Malignant glioma (MG) is a devastating neuro-oncologic disease with almost invariably poor prognosis, yet many families facing malignant glioma have poor prognostic awareness (PA), or the awareness of the patient's incurable disease and shortened life expectancy. Accurate PA is associated with favorable medical outcomes at end-of-life for patients and psychosocial outcomes for informal caregivers (ICs) through bereavement. To date, however, no study has specifically examined PA among MG ICs and the information they receive that shapes their awareness. METHODS: Thirty-two ICs of patients with malignant glioma completed a semi-structured assessment of their awareness of the incurability and life expectancy of their loved one's illness, and to understand their sources of prognostic information and preferences for communication of prognostic information. RESULTS: Twenty-two (69%) ICs had full PA-awareness of the incurability of malignant glioma and accurate estimates of their loved ones' life expectancy. Twenty-three (72%) felt that prognostic information was extremely or very important to possess, and 16 (50%) desired more prognostic information. The majority of ICs received prognostic information from physicians and the Internet. Qualitative analyses revealed that many ICs had difficulty navigating medical encounters in which they concurrently wanted to elicit prognostic information from physicians and protect patients from such information. CONCLUSIONS: Accurate and timely PA is necessary for ICs to serve as critical members of health care teams. Interventions are needed to foster ICs' skills in navigating prognostic communication with patients and health care providers and thereby improve their ability to advocate for their loved one's wishes.

The Common Sense of Counseling Psychology: Introducing the Common-Sense Model of Self-Regulation.

The goal of therapy is typically to improve clients' self-management of their problems, not only during the course of therapy but also after therapy ends. Although it seems obvious that therapists are interested in improving client's self-management, the psychotherapy literature has little to say on the topic. This article introduces Leventhal's Common-Sense Model of Self-Regulation, a theoretical model of the self-management of health, and applies the model to the therapeutic process. The Common-Sense Model proposes that people develop illness representations of health threats and these illness representations guide self-management. The model has primarily been used to understand how people self-manage physical health problems, we propose it may also be useful to understand self-management of mental health problems. The Common-Sense Model's strengths-based perspective is a natural fit for the work of counseling psychologists. In particular, the model has important practical implications for addressing how clients understand mental health problems over the course of treatment and self-manage these problems during and after treatment.

Smoking and IPMN malignant progression.

BACKGROUND: Patients with intraductal papillary mucinous neoplasm (IPMN) are at risk for invasive pancreatic cancer. We aim to characterize the impact of smoking on IPMN malignant progression. METHODS: Patients undergoing pancreatic resection for IPMN (1991-2015) were retrospectively reviewed using a prospectively collected database. RESULTS: Of 422 patients identified, 324 had complete data for analysis; 55% were smokers. Smoking status did not impact IPMN malignant progression (smokers/non-smokers: 22%/18% invasive grade; p = 0.5). Smokers were younger than non-smokers at the time of IPMN diagnosis (63 versus 68 years; p = 0.001). This association also held in the invasive IPMN subgroup (65 versus 72 years, p = 0.01). Despite this observation, rate of symptoms at diagnosis, cancer stage, and median survival were the same between smokers and non-smokers. CONCLUSION: Although smoking is not associated with IPMN malignant progression, invasive IPMN is diagnosed at a younger age in smokers. These data suggest tobacco exposure may accelerate IPMN malignant progression.

Residual feed intake, body composition, and fertility in yearling beef heifers.

One hundred thirty-seven spring-born yearling beef heifers of British breed types were used to determine the relationships between residual feed intake (RFI) and growth rate, body composition, mature size, and fertility. Heifers were housed in a dry lot facility during the experimental period, and data were collected over a 2-yr period (yr 1, n = 67; yr 2, n = 70). Individual feed intake, BW, BCS, hip height, and ultrasonic measurements [subcutaneous rib fat (UBF), rump fat (URF), LM area (LMA), and intramuscular fat (IMF)] of body composition were recorded. Individual feed intakes (kg of TDN consumed/d) were used to calculate RFI combining both years of data. Heifers averaged 387.0 ± 19.4 d of age and 337.1 ± 29.9 kg of BW at initiation of the experiment. Mean ADG was 1.14 ± 0.21 kg/d during the trial. Based on RFI, with year of test and farm of origin included in the model as covariates, heifers were classified into groups: positive (POS; 0.74 kg of TDN/d) or negative (NEG; -0.73 kg TDN/d) for first analysis and high (HI), medium (MED), or low (LO; mean RFI = 1.06, -0.01, and -1.13 kg of TDN/d, respectively) subsequently. An initial phenotypic relationship (P < 0.05) between RFI and both UBF and URF (r = 0.19 and 0.17, respectively) was sustained (P < 0.01) with UBF (r = 0.27) and URF (r = 0.24) to trial conclusion. No other correlations with RFI were significant. Heifers classified as POS reached puberty earlier than those classified as NEG (414 ± 3.83 vs. 427 ± 4.67 d of age, P = 0.03), and possessed greater LMA per 100 kg of BW (LMACWT) at conclusion of the trial (P < 0.01). Medium heifers exhibited less URF (P < 0.05) compared with either HI or LO heifers at trial initiation. Low heifers possessed less LM area (cm(2)) per 100 kg of BW (P < 0.05) than HI but did not differ (P > 0.10) from MED heifers at either the beginning or the end of test. Additionally, a negative linear relationship was observed between RFI and age at puberty (P < 0.05). Each 1-unit increase in RFI corresponded to a decrease of 7.5 d in age at puberty, but did not affect pregnancy or conception rates (P > 0.10). Differences in body fat and rate of metabolism associated with RFI could delay reproductive maturity.

Evaluation of temephos formulations against Aedes taeniorhynchus larvae.

Four commercially available temephos formulations (Abate 4E, Clarke 1% Skeeter Abate [1% SA], Abate 2-BG, and Abate 5-BG) were evaluated against 3rd instars of Aedes taeniorhynchus in laboratory beaker tests and small field test plots. Abate 4E liquid formulation was effective in both the laboratory and plot tests. 1% SA and Abate 5-BG were more effective in the small field test plots than Abate 2-BG.

Efficacy studies of Aquaprene (1.8% and 2.8% AI) sand granules and Altosid XR-G (1.5% AI) sand granules against first and second instars of Ochlerotatus taeniorhynchus as a preflood treatment in small field plots.

An efficacy study was conducted to evaluate sand granule formulations of Aquaprene (1.8% and 2.8% active ingredient [AI]) and Altosid XR-G (1.5% AI) as a preflood application against Ochlerotatus taeniorhynchus larvae in small field test plots. Aquaprene sand granules (2.8% Al) were applied at 2.5 and 5 lb/acre and the 1.8% AI formulation at 4.2 lb/acre. The 1.8% AI formulation was compared with Altosid XR-G sand granules (1.5% AI) applied at 5 lb/acre. Plots were flooded after 7 days, and 1st and 2nd instars were introduced, pupae were collected, and the plots were drained and dried for 9 days. Assessments were made 21 and 35 days posttreatment. Both Aquaprene sand granules formulations exhibited excellent control throughout the 35-day study.

Efficacy studies of Aquaprene (emulsifiable concentrate and wettable powder) insect growth regulator formulations against third- and fourth-stage larvae of Ochlerotatus taeniorhynchus in small-plot field studies.

Efficacy studies were conducted with Aquaprene emulsifiable concentrate (EC) (33.6% active ingredient [AI]) and wettable powder (WP) (40% AI) (S)-methoprene insect growth regulator formulations against larval Ochlerotatus taeniorhynchus in small field test plots. Aquaprene EC was applied at 7.18 and 9.57 g/acre. Approximately one thousand 4th-stage larvae were added to each plot before treatment, and 24 h later pupae were collected to determine emergence inhibition. At both applications rates, Aquaprene EC was extremely effective (99%) at significantly reducing adult emergence in the studies. Two application rates ((S)-methoprene at 2.4 and 4.8 g/acre) of the Aquaprene WP also were evaluated against Oc. taeniorhynchus at 1, 3, 7, and 14 days after treatment. Emergence inhibition and statistical analysis showed that significant differences were found between the (S)-methoprene rates of 2.4 and 4.8 g/acre at each posttreatment assessment. The (S)-methoprene rate of 4.8 g/acre was effective in controlling adult emergence for up to 14 days after treatment.

Efficacy studies of Vectobac 12as and Teknar HP-D larvicides against 3rd-instar Ochlerotatus taeniorhynchus and Culex quinquefasciatus in small plot field studies.

Efficacy studies were conducted with VectoBac 12AS and Teknar HP-D larvicides against 3rd-instar Ochlerotatus taeniorhynchus and Culex quinquefasciatus in small field test plots. The products were obtained off the shelf from distributors and had different lot numbers. They were evaluated over a 2-year period in spring 2002 and 2003. Application rates were 0.29, 0.58, and 1.10 liter/ha and evaluations were made 24 and 48 h after treatment. Both products performed well in these studies, with VectoBac 12AS being more effective at the 0.29 liter/ha rate.

Dynamic infrared imaging of newly diagnosed malignant lymphoma compared with Gallium-67 and Fluorine-18 fluorodeoxyglucose (FDG) positron emission tomography.

Staging and therapy monitoring of malignant lymphomas relies heavily on imaging using arbitrary size criteria from computed tomography (CT) and sometimes non-specific radionuclide studies to assess the activity of the disease. Treatment decisions are based on early assessment of the response to therapy and the residual volume of the disease. Our initial experience is reported using a new noninvasive, inexpensive, and reproducible passive imaging modality, Dynamic Infrared Imaging (DIRI), which may add a new dimension to functional imaging. This system relies on its ability to filter the raw infrared signal using biological oscillatory behavior. It detects and analyzes minute oscillations of temperature and heat distribution in tumors.

Acute exposure of toluene transiently potentiates p42/44 mitogen-activated protein kinase (MAPK) activity in cultured rat cortical astrocytes.

It has been shown that the inhalation of toluene in rats can cause neuronal apoptosis in the central nervous system. However, the cellular and molecular effects of toluene directly on astrocytes are relatively unknown. We used primary cultures of astrocytes isolated from the neonatal rat cortex as a model to study the toluene effects on cell outcome and associated signal transduction pathways using immunostaining and Western blotting. We observed that acute toluene exposure significantly induced caspase-dependent cell apoptosis and transiently stimulated the activation of p42/44 mitogen-activated protein kinase (MAPK) in the primary astrocytes. Interestingly, the inhibition of the p42/44 MAPK signaling cascade by PD98059 in conjunction with the toluene treatment evoked more cellular apoptosis than toluene alone, suggesting that the toluene-induced transient MAPK activation may play a role in promoting cell survival during the toluene exposure.

Cytotoxicity of 2-aldo- and 2-ketopyridine-N(4)-substituted thiosemicarbazones and mode of action in human Tmolt4 cells.

The 2-aldo- and 2-ketopyridine-N(4)-substituted thiosemicarbazones and their copper complexes demonstrated potent cytotoxic activity against a series of murine and human suspended cultured tumor cells. Selected compounds were active against the growth of cultured cells from solid human tumors, i.e. Mck-7 breast effusion, lung A549 and lung MB-9812, bone SOS-2 and clear cell Caki renal tumor. In Tmolt4 T cell leukemia cells the compounds inhibited the syntheses of DNA, RNA and protein over 60 min at 25 to 100 microM. Multiple target sites in nucleic acid metabolism were suppressed by the agents, i.e. DNA polymerase alpha, ribonucleoside reductase, dihydrofolate reductase, de novo purine synthesis, thymidylate synthetase and nucleoside kinases. The total effects of the agents on DNA metabolism led to the reduction of deoxyribonucleotide pools as well as DNA fragmentation.

The use of GABA(A) receptors expressed in neural precursor cells for cell-based assays.

GABA(A) receptors are known targets for certain classes of environmental neurotoxins and pharmaceutical compounds. Since few neural cell lines express functional GABA(A) receptors, the capacity to rapidly screen for compounds that affect GABA(A) receptor function is presently limited. Previous work has demonstrated that rat neural precursor cells express functional GABA(A) receptors that can be monitored via Ca(2+) imaging. This study examined GABA(A) receptor subunit expression to determine whether GABA(A) receptor function and its interactions with neurotoxins is preserved after passaging. Neural precursor cells isolated from embryonic day 13 rat brain were expanded in serum-free medium containing basic fibroblast growth factor and passaged three times. Reverse transcription-polymerase chain reaction analysis demonstrated early expression of abundant mRNAs encoding various GABA(A) receptor subunits. Ca(2+) imaging showed that the highly proliferating precursor cells in passaged cultures maintained expression of functional GABA(A) receptors. In addition, we showed that trimethylolpropane phosphate, a neurotoxin generated during partial pyrolysis of a synthetic ester turbine engine lubricant, potently inhibited muscimol (GABA(A) receptor agonist) but not depolarization-induced cytosolic Ca(2+) increase. The findings of this study suggest that neural precursor cells may be well suited for the evaluation of certain environmental neurotoxins with convulsant activity. The potential use of neural precursor cells in high-throughput screens for compounds acting on GABA(A) receptors is discussed.

Design and demonstration of an automated cell-based biosensor.

Cell-based biosensors have the capacity to respond to a wide range of analytes in a physiologically relevant manner and appear well-suited for toxicity monitoring of both known and unknown analytes. One means of acquiring cellular functional information for biosensor applications involves extracellular recording from excitable cells, which can generate noninvasive and long-term measurements. Previous work from our laboratory described a prototype portable system capable of high signal-to-noise extracellular recordings, in spite of deficiencies in thermal control, fluidics handling, and absence of data acquisition (DAQ) capability. The present work describes a cell-based biosensor system that incorporates low noise amplifier and filter boards, a two-stage thermal control system with integrated fluidics and a flexible graphical user interface for DAQ and control implemented on a personal computer. Wherever possible, commercial off-the-shelf components have been utilized for system design and fabrication. The system exhibits input-referred noise levels of 5-10 microV(RMS), such that extracellular potentials exceeding 50-60 microV can be readily resolved. In addition, the biosensor system is capable of automated temperature and fluidics control. Flow rates can range from 0-2.5 ml/min, while the cell recording chamber temperature is maintained within a range of 36-37 degrees C. To demonstrate the capability of this system to resolve small extracellular potentials, recordings from embryonic chick cardiac myocytes have been performed.

Targeting of human Tmolt4 leukemic type II IMP dehydrogenase by cyclic imide related derivatives.

2,3-Dihydrophthalazine-1,4-diones, indazolones, 3-imino-1-oxoisodolines, homophthalimides, napthalidimides, diphenamides, and 6,7-dihydro-5H-dibenz[c,e]azepines proved to be potent inhibitors of the activity of human Tmolt4 T cell leukemia Type II IMP dehydrogenase (IMPDH). This inhibition was competitive, yielding Ki values in the range of 1.96 to 48.9 microM. The inhibition of Type II IMPDH correlated positively with the inhibition of the growth of Tmolt4 cells, the syntheses of DNA and purine, and the activity of crude IMPDH. The Type II IMPDH isoform is found in rapidly proliferating cells. The isoform present in normal resting cells, Type I IMPDH, was elevated by the compounds at 100 microM. In addition, Compound 5 significantly increased the Type I enzyme activity in a concentration and time dependent manner. The selectivity of these derivatives towards Type II IMPDH will allow for the separation of cellular effects, which should reduce clinical toxicity when treating with antimetabolite IMPDH inhibitors.

Detection of physiologically active compounds using cell-based biosensors.

Cell-based biosensors are portable devices that contain living biological cells that monitor physiological changes induced by exposure to environmental perturbations such as toxicants, pathogens or other agents. Methods of detecting physiological changes include extracellular electrical recordings, optical measurements, and, in the future, functional genomics and proteomics. Several technical developments are occurring that will increase the feasibility of cell-based biosensors for field applications; these developments include stem cell and 3D culture technologies. Possible scenarios for the use of cell-based biosensors include broad-range detectors of unknown threat agents and functional assessment of identified agents.

Ethanol blocks cytosolic Ca2+ responses triggered by activation of GABA(A) receptor/Cl- channels in cultured proliferating rat neuroepithelial cells.

GABA(A) receptor/Cl- channels and voltage-gated Ca2+ channels are believed to be important sites of ethanol action in the CNS. Acute exposure of ethanol potentiates GABA(A) receptor/Cl- channel activity and inhibits voltage-gated Ca2+ channels in a number of preparations, mostly post-mitotic neurons. The effects of ethanol on these channels in primary cultures of undifferentiated neural precursor cells remain unknown. To address this issue, we examined the effects of ethanol on GABA(A) agonist-activated elevation of cytosolic Ca2+ in an in vitro model of the cortical neuroepithelium derived from rat basic fibroblast growth factor-expanded neural precursor cells. We found a potent inhibition of GABA(A)-activated elevation of cytosolic Ca2+ by ethanol in actively proliferating cells. Since we had recently demonstrated that GABA(A) receptor activation depolarizes these cells and elevates their cytosolic Ca2+, we tested whether the effects of ethanol involved both GABA(A) receptors and voltage-gated Ca2+ channels. Both extracellular K+- and muscimol-induced cytosolic Ca2+ elevations were abolished by nitrendipine, indicating that both depolarizing stimuli triggered Ca2+ influx through L-type voltage-gated Ca2+ channels. Exposure of proliferating cells to different concentrations of ethanol revealed that the drug was more potent in blocking muscimol-induced compared to K+-evoked cytosolic Ca2+ elevations. These results raise the possibility that ethanol blocks GABAergic stimulation of cytosolic Ca2+ levels in proliferating precursors primarily by interacting with GABA(A) receptor/Cl- channels and secondarily with voltage-gated Ca2+ channels.

Efficacy of ground-applied ultra-low-volume malathion on honey bee survival and productivity in open and forest areas.

A study was conducted to determine the efficacy of ground ultra-low-volume malathion sprays on honey bee (Apis mellifera L.) apiaries in open and forested areas downwind from the spray route. Impact on colonies 7.6, 15.2, 47.7, and 91.4 m downwind from sprays was assessed by recording individual bee mortality 12 and 36 h after treatment. In addition, hives were weighed before as well as during the study and cluster counts were conducted at each hive to determine colony strength before and after treatment. Spray drift was monitored by the use of caged mosquito (Culex quinquefasciatus Say) mortality and deposition on filter paper. During the study, significant bee mortality in the open area occurred on 2 occasions at 7.6 m (16.8 +/- 4.3 bees, 11.8 +/- 7.0 bees) and at 15.2 m (6.5 +/- 1.7 bees, 5.3 +/- 1.5 bees). Significant mortality in the forested area was observed only once and consisted of 2 bees at 7.6 m. In each case where bee mortality occurred, spray deposits on filter papers had exceeded 400 ng/cm2. Although mortality of caged mosquitoes indicated that malathion drifted through the study areas, little correlation was apparent between mortality and spray deposition on filter paper.

Survival and neurite outgrowth of rat cortical neurons in three-dimensional agarose and collagen gel matrices.

To better understand interactions between neurons and extracellular matrix equivalents, embryonic day-18 rat cortical neurons were immobilized and maintained in culture for up to 24 days in agarose and type I collagen gels. Using live/dead staining, neuronal cultures in low density collagen gel lasted at least 3 weeks. At 14 days, over 50% of immobilized cells in collagen gel were found viable while in low density agarose gel no cells survived. In situ cell death detection showed that most, if not all, dead cells in either of the gels underwent apoptosis. The collagen-trapped neurons exhibited normal neuronal polarity and developed long neurites, estimated at over 500 microm. The results suggest that collagen, because it is a major extracellular matrix constituent, suppresses apoptosis and provides a suitable substrate for neuronal survival and differentiation.

Clinical competence assessment in radiology: introduction of an objective structured clinical examination in the medical school curriculum.

RATIONALE AND OBJECTIVES: Traditional oral and written examinations can be limited in predicting future clinical performance. Therefore Objective Structured Clinical Examinations (OSCEs) have been introduced in other specialties. The authors assessed their value in radiology. MATERIALS AND METHODS: The study includes 122 Harvard medical students who undertook 1-month compulsory clerkships at one of three hospitals (A, B, or C) in their 3rd and 4th year and a compulsory OSCE in their 4th year. The OSCE was constructed from five cases. Each had eight or nine standardized questions designed to test, within a set time, the perception of essential findings, their interpretation, and clinical judgment (maximum possible score, 100). Clerkship grades were high honors (score of 3), honors (score of 2), satisfactory (score of 1), and fail (score of 0). Predictors of OSCE scores-clerkship grade and affiliated hospital-were modeled as linear functions. Time elapsed between clerkship and OSCE was modeled as a nonlinear function. RESULTS: Although there was a positive relation between clerkship grade and OSCE grade, it accounted for an increase of only 5.7% in OSCE score per clerkship grade and did not predict performance of individual students. Students who trained in hospital B showed significantly higher OSCE grades. OSCE scores were highest when the examination was taken 8 months after the clerkship. CONCLUSION: The OSCE may be useful to uncover deficits in individuals and groups beyond the ones detected with traditional clerkship evaluations and provide guidance for remediation. The improved performance after additional clinical exposure suggests that the OSCE may be well suited to test the integration of radiologic and clinical knowledge.