The MLL-Menin Interaction is a Therapeutic Vulnerability in NUP98-rearranged AML.

Chromosomal translocations involving the NUP98 locus are among the most prevalent rearrangements in pediatric acute myeloid leukemia (AML). AML with NUP98 fusions is characterized by high expression of HOXA and MEIS1 genes and is associated with poor clinical outcome. NUP98 fusion proteins are recruited to their target genes by the mixed lineage leukemia (MLL) complex, which involves a direct interaction between MLL and Menin. Here, we show that therapeutic targeting of the Menin-MLL interaction inhibits the propagation of NUP98-rearrranged AML both ex vivo and in vivo. Treatment of primary AML cells with the Menin inhibitor revumenib (SNDX-5613) impairs proliferation and clonogenicity ex vivo in long-term coculture and drives myeloid differentiation. These phenotypic effects are associated with global gene expression changes in primary AML samples that involve the downregulation of many critical NUP98 fusion protein-target genes, such as MEIS1 and CDK6. In addition, Menin inhibition reduces the expression of both wild-type FLT3 and mutated FLT3-ITD, and in combination with FLT3 inhibitor, suppresses patient-derived NUP98-r AML cells in a synergistic manner. Revumenib treatment blocks leukemic engraftment and prevents leukemia-associated death of immunodeficient mice transplanted with NUP98::NSD1 FLT3-ITD-positive patient-derived AML cells. These results demonstrate that NUP98-rearranged AMLs are highly susceptible to inhibition of the MLL-Menin interaction and suggest the inclusion of AML patients harboring NUP98 fusions into the clinical evaluation of Menin inhibitors.

Intranasal deferoxamine engages multiple pathways to decrease memory loss in the APP/PS1 model of amyloid accumulation.

In addition to the hallmark accumulation of amyloid and hyper-phosphorylation of tau, brain changes in Alzheimer's disease are multifactorial including inflammation, oxidative stress, and metal dysregulation. Metal chelators have been explored as a less well known approach to treatment. One chelator currently being developed is deferoxamine (DFO), administered via the intranasal (IN) route. In the current study, APP/PS1 amyloid mice were treated with a chronic, low dose of IN DFO, subjected to a rigorous battery of behavior tests, and the mechanism of action was examined. Mice were treated 3x/week with 0.24 C IN DFO for 18 weeks from 36 to 54 weeks of age, 4 weeks of behavior tests were performed that included both working and reference memory, anxiolytic and motor behaviors, and finally brain tissues were analyzed for amyloid, protein oxidation, and other proteins affected by DFO. We found that IN DFO treatment significantly decreased loss of both reference and working memory in the Morris and radial arm water mazes (p < 0.05), and also decreased soluble Aß40 and Aß42 in cortex and hippocampus (p < 0.05). Further, IN DFO decreased activity of GSK3ß, and led to decreases in oxidative stress (p < 0.05). These data demonstrate that low doses of IN DFO can modify several targets along the multiple pathways implicated in the neuropathology of Alzheimer's, making it an attractive candidate for the treatment of this heterogeneous disease.

In search of compassion: a new taxonomy of compassionate physician behaviours.

BACKGROUND: Compassion has been extolled as a virtue in the physician-patient relationship as a response to patient suffering. However, there are few studies that systematically document the behavioural features of physician compassion and the ways in which physicians communicate compassion to patients. OBJECTIVE: To develop a taxonomy of compassionate behaviours and statements expressed by the physician that can be discerned by an outside observer. DESIGN: Qualitative analysis of audio-recorded office visits between oncologists and patients with advanced cancer. SETTING AND PARTICIPANTS: Oncologists (n = 23) and their patients with advanced cancer (n = 49) were recruited in the greater Rochester, New York, area. The physicians and patients were surveyed and had office visits audio recorded. MAIN OUTCOME MEASURES: Audio recordings were listened to for qualitative assessment of communication skills. RESULTS: Our sensitizing framework was oriented around three elements of compassion: recognition of the patient's suffering, emotional resonance and movement towards addressing suffering. Statements of compassion included direct statements, paralinguistic expressions and performative comments. Compassion frequently unfolded over the course of a conversation rather than being a single discrete event. Additionally, non-verbal linguistic elements (e.g. silence) were frequently employed to communicate emotional resonance. DISCUSSION AND CONCLUSIONS: This study is the first to systematically catalogue instances of compassionate communication in physician-patient dialogues. Further refinement and validation of this preliminary taxonomy can guide future education and training interventions to facilitate compassion in physician-patient interactions.

Movement of cyantraniliprole in plants after foliar applications and its impact on the control of sucking and chewing insects.

BACKGROUND: Given the physical properties of insecticides, there is often some movement of these compounds within crop plants following foliar application. In this context, movement of two formulations of cyantraniliprole, an anthranilic diamide, was characterized for translocation to new growth, distribution within a leaf and penetration through the leaf cuticle. RESULTS: Upward movement of cyantraniliprole to new plant growth via the xylem was confirmed using (14) C-radiolabeled cyantraniliprole and from Helicoverpa zea mortality on tomato leaves that had not been directly treated. Within a leaf there was significant acropetal movement (base to apex) of cyantraniliprole, but no significant basipetal movement (apex to base). Translaminar movement, the ability of a compound to penetrate the leaf cuticle, was demonstrated in a variety of plants, both with and without the use of adjuvants, by treating only the adaxial surface of the leaf and measuring control of diamondback moth (Plutella xylostella), green peach aphid (Myzus persicae) and sweetpotato whitefly (Bemisia tabaci) exposed in clip cages to the untreated abaxial surface. CONCLUSION: The plant mobility and plant protection of cyantraniliprole is discussed with implications for use in insect resistance management and integrated pest management programs.

"Speaking-for" and "speaking-as": pseudo-surrogacy in physician-patient-companion medical encounters about advanced cancer.

OBJECTIVE: To examine using audio-recorded encounters the extent and process of companion participation when discussing treatment choices and prognosis in the context of a life-limiting cancer diagnosis. METHODS: Qualitative analysis of transcribed outpatient visits between 17 oncologists, 49 patients with advanced cancer, and 34 companions. RESULTS: 46 qualifying companion statements were collected from a total of 28 conversations about treatment choices or prognosis. We identified a range of companion positions, from "pseudo-surrogacy" (companion speaking as if the patient were not able to speak for himself), "hearsay", "conflation of thoughts", "co-experiencing", "observation as an outsider", and "facilitation". Statements made by companions were infrequently directly validated by the patient. CONCLUSION: Companions often spoke on behalf of patients during discussions of prognosis and treatment choices, even when the patient was present and capable of speaking on his or her own behalf. PRACTICE IMPLICATIONS: The conversational role of companions as well as whether the physician checks with the patient can determine whether a companion facilitates or inhibits patient autonomy and involvement. Physicians can reduce ambiguity and encourage patient participation by being aware of when and how companions may speak on behalf of patients and by corroborating the companion's statement with the patient.

Use of honeydew production to determine reduction in feeding by Bemisia tabaci (Hemiptera: Aleyrodidae) adults when exposed to cyantraniliprole and imidacloprid treatments.

Bemisia tabaci (Gennadius) whitefly populations produce economically important damage to crops by their direct feeding and also by transmitting plant viruses. Although there are several methods to reduce B. tabaci damage, most growers rely on the use of insecticides to decrease populations of this pest. Insecticides that reduce feeding of whitefly adults may also reduce the transmission of viruses that are harmful to crop plants. However, demonstrating the feeding reduction has proved challenging. In this study, water-sensitive paper was used to determine whitefly adult feeding, indirectly through honeydew production, when insects were placed on insecticide-treated and untreated plants. Plant treatments with two formulations of cyantraniliprole (Cyazypyr) showed a reduction in the amount of honeydew produced by B. tabaci adults equivalent to imidacloprid. The reduction in the amount of honeydew produced indicates reduced insect feeding and the possibility for a reduction in virus transmission. Plant treatments with two formulations of cyantraniliprole also resulted in higher mortality than imidacloprid.

Use of fluorescence, a novel technique to determine reduction in Bemisia tabaci (Hemiptera: Aleyrodidae) nymph feeding when exposed to Benevia and other insecticides.

The sweet potato whitefly, Bemisia tabaci (Gennadius), is an economically important pest in the United States and other countries. Growers in many places rely on the use of insecticides to reduce populations of B. tabaci. However, insecticides may take a few days to cause B. tabaci mortality and some do not reduce feeding before death. Earlier reduction of feeding of whiteflies would decrease the physiological effects on plants, reduce the production of sooty mold and potentially reduce the transmission of viruses. Measuring the reduction in feeding after the exposure of B. tabaci to an insecticide has proven difficult. This series of laboratory experiments demonstrate the usefulness of fluorescence in determining B. tabaci feeding cessation. Fluorescein sodium salt is systemically transported in the xylem from the roots to the plant leaves and absorbed by B. tabaci nymphs feeding on these plants. Nymphs start fluorescing shortly after the cotton plant root system is submerged in the fluorescein sodium salt. Using this novel technique, the effect of three insecticides with different modes of action, cyantraniliprole, imidacloprid, and spirotetramat on B. tabaci was evaluated and compared to determine reduction in feeding. Results indicate that B. tabaci nymphs feeding on a plant treated with Benevia have a significant reduction of feeding when compared with nymphs feeding on plants treated with imidacloprid or spirotetramat. Both Benevia and spirotetramat caused significant nymphal mortality by 48 h after exposure. This novel technique will be useful to demonstrate the feeding cessation or reduction in feeding produced by different insecticides in several sucking insect groups.

A resource for the conditional ablation of microRNAs in the mouse.

The importance of miRNAs during development and disease processes is well established. However, most studies have been done in cells or with patient tissues, and therefore the physiological roles of miRNAs are not well understood. To unravel in vivo functions of miRNAs, we have generated conditional, reporter-tagged knockout-first mice for numerous evolutionarily conserved miRNAs. Here, we report the generation of 162 miRNA targeting vectors, 64 targeted ES cell lines, and 46 germline-transmitted miRNA knockout mice. In vivo lacZ reporter analysis in 18 lines revealed highly tissue-specific expression patterns and their miRNA expression profiling matched closely with published expression data. Most miRNA knockout mice tested were viable, supporting a mechanism by which miRNAs act redundantly with other miRNAs or other pathways. These data and collection of resources will be of value for the in vivo dissection of miRNA functions in mouse models.

Sustained antigen-specific antitumor recall response mediated by gene-modified CD4+ T helper-1 and CD8+ T cells.

Given that specific subsets of T helper 1 (Th1) and T helper 2 (Th2) CD4(+) T cells have been shown to play key roles in tumor rejection models, we wanted to assess the contribution of either Th1 or Th2 CD4(+) cell subtypes for redirected T-cell immunotherapy. In this study, we have developed a novel method involving retroviral transduction and in vitro T-cell polarization to generate gene-engineered mouse CD4(+) Th1 and Th2 cells or T helper intermediate (Thi) cells expressing an anti-erbB2-CD28-zeta chimeric receptor. Gene-modified Th1 and Th2 polarized CD4(+) cells were characterized by the preferential secretion of IFN-gamma and interleukin-4, respectively, whereas Thi cells secreted both cytokines following receptor ligation. In adoptive transfer studies using an erbB2(+) lung metastasis model, complete survival of mice was observed when transduced Th1, Th2, or Thi CD4(+) cells were transferred in combination with an equivalent number of transduced CD8(+) T cells. Tumor rejection was consistently associated with transduced T cells at the tumor site and interleukin-2 secretion. However, the surviving mice treated with gene-modified Th1 CD4(+) cells were significantly more resistant to a subsequent challenge with a different erbB2(+) tumor (4T1.2) implanted s.c. This result correlated with both increased expansion of Th1 CD4(+) and CD8(+) T cells in the blood and a greater number of these cells localizing to the tumor site following rechallenge. These data support the use of gene-modified CD4(+) Th1 and CD8(+) T cells for mediating a sustained antitumor response.

Image-based high-throughput quantification of cellular fat accumulation.

A number of biochemical methods are available for measuring fat accumulation in cell culture. The authors report a simple image-based method for measuring fat accumulation in adipocytes using a combination of high-throughput brightfield microscopy and image analysis, which was validated biochemically using Oil-Red-O. The quickest and most accurate method of analysis was one based on thresholding brightfield images and determining the area of fat droplets per image. Thus, the authors have developed a simple high-throughput, label-free method for measuring fat accumulation that is applicable to any cell or tissue type where fat droplets are visible under light microscopy.

Communicating risk information: the influence of graphical display format on quantitative information perception-Accuracy, comprehension and preferences.

OBJECTIVE: Statistical health risk information has proved notoriously confusing and difficult to understand. While past research indicates that presenting risk information in a frequency format is superior to relative risk and probability formats, the optimal characteristics of frequency formats are still unclear. The aim of this study is to determine the features of 1000 person frequency diagrams (pictographs) which result in the greatest speed and accuracy of graphical perception. METHODS: Participants estimated the difference in chance of survival when taking or not taking Drug A, on a pictograph format, varying by mode (one-graph/two-graph), direction (vertical/horizontal), and shading (shaded/unshaded), and their preferences for the different formats. Their understanding of different components of the 1000 person diagram was assessed. Responses were timed and scored for accuracy. RESULTS: Horizontal pictographs were perceived faster and more accurately than vertical formats. Two-graph pictographs were perceived faster than one-graph formats. Shading reduced response time in two-graph formats, but increased response times in one-graph formats. Shaded and one-graph pictographs were preferred. CONCLUSIONS: As shading and one-graph formats were preferred, further clarification as to why shading negatively impacts on response times in the one-graph format is warranted. PRACTICE IMPLICATIONS: Horizontal pictographs are optimal.

Effect of referral source on outcome with cognitive-behavior therapy self-help.

Little is known about how psychiatric patients' source of referral relates to treatment outcome. This study examines the effect of referral source on clinical outcome with computer-aided cognitive-behavior therapy (CCBT) for anxiety and depressive disorders. Three hundred fifty-five referrals to a clinic that offered CCBT with brief backup from a clinician were classified into general practitioner (GP) referrals (34%), mental health (MH) professional referrals (42%), and self-referrals (SR, 24%), and compared on sociodemographic and clinical features and treatment outcome. At intake, referrals from all 3 sources had similar sociodemographic features and problem duration, but GP referrals had less comorbidity, whereas MH professional referrals were being treated for their problem more often and were less motivated to change than were SR. Among treatment completers, SRs had the least and MH professional referrals had the most impaired work/social adjustment. Each referral group improved on generic and syndrome-specific measures; however, GP referrals improved the most and MH professional referrals the least. The 3 groups received similar therapist support and were equally satisfied after treatment. We conclude that GP referrals had the best outcome with CCBT for anxiety/depressive disorders. Referral source can be important in psychotherapy research because it may affect the type of patient seen and may predict treatment outcome.

Reliability and validity of the work and social adjustment scale in phobic disorders.

The Work and Social Adjustment Scale (WSAS) is a simple widely used 5-item measure of disability whose psychometric properties need more analysis in phobic disorders. The reliability, factor structure, validity, and sensitivity to change of the WSAS were studied in 205 phobic patients (73 agoraphobia, 62 social phobia, and 70 specific phobia) who participated in various open and randomized trials of self-exposure therapy. Internal consistency of the WSAS was excellent in all phobics pooled and in agoraphobics and social phobics separately. Principal components analysis extracted a single general factor of disability. Specific phobics gave less consistent ratings across WSAS items, suggesting that some items were less relevant to their problem. Internal consistency was marginally higher for self-ratings than clinician ratings of the WSAS. Self-ratings and clinician ratings correlated highly though patients tended to rate themselves as more disabled than clinicians did. WSAS total scores reflected differences in phobic severity and improvement with treatment. The WSAS is a valid, reliable, and change-sensitive measure of work/social and other adjustment in phobic disorders, especially in agoraphobia and social phobia.

2-Ethoxybenzoxazole as a bioisosteric replacement of an ethyl benzoate group in a human rhinovirus (HRV) capsid binder.

A series of pyridazinylpiperidinyl capsid-binding compounds with novel bicyclic substituents were synthesized and screened against human rhinovirus (HRV). Several 2-alkoxy- and 2-alkylthio-benzoxazole and benzothiazole derivatives showed excellent anti-HRV activity. When tested against a panel of 16 representative HRV types the 2-ethoxybenzoxazole derivative 13 was found to have superior HRV activity (median EC(50) 3.88ng/mL) to known capsid-binders Pleconaril and Pirodavir. Compound 13 illustrates that a 2-alkoxybenzoxazole group can be an effective bioisostere for a benzoate ester or benzaldehyde oxime ether functionality.

Dimeric zanamivir conjugates with various linking groups are potent, long-lasting inhibitors of influenza neuraminidase including H5N1 avian influenza.

The synthesis, antiviral and pharmacokinetic properties of zanamivir (ZMV) dimers 8 and 13 are described. The compounds are highly potent neuraminidase (NA) inhibitors which, along with dimer 3, are being investigated as potential second generation inhaled therapies both for the treatment of influenza and for prophylactic use. They show outstanding activity in a 1 week mouse influenza prophylaxis assay, and compared with ZMV, high concentrations of 8 and 13 are found in rat lung tissue after 1 week. Retention of compounds in rat lung tissue correlated both with molecular weight (excluding 3 and 15) and with a capacity factor K' derived from immobilized artificial membrane (IAM) chromatography (including 3 and 15). Pharmacokinetic parameters for 3, 8 and 13 in rats show the compounds have short to moderate plasma half-lives, low clearances and low volumes of distribution. Dimer 3 shows NA inhibitory activity against N1 viruses including the recent highly pathogenic H5N1 A/Chicken/Vietnam/8/2004. In plaque reduction assays, 3, 8 and 13 show good to outstanding potency against a panel of nine flu A and B virus strains. Consistent with its shorter and more rigid linking group, dimer 8 has been successfully crystallized.

Screening people with anxiety/depression for suitability for guided self-help.

The aim of this study was to test a questionnaire for screening people with anxiety/ depression for their suitability for certain forms of computer-guided self-help. A total of 196 referrals completed the screening questionnaire. Three clinicians each independently judged the referrals' broad problem type and suitability. Referrals were randomized to 1 of 3 clinicians for a screening interview. The results show that inter-clinician agreement was good for questionnaire-based problem type and suitability, and excellent for screening interview-based problem type and suitability. Agreement between the questionnaire and interview was good on problem type but poor on suitability. Compared with the screening interview, the questionnaire detected suitable patients well but unsuitable patients less well. In conclusion, by quickly scanning the completed questionnaire, clinicians were able sensitively to detect patients' problem types that were suitable for certain forms of self-help. Some unsuitability items need refining.

Investigations into the cellular actions of the shellfish toxin gymnodimine and analogues.

The effects of the shellfish toxin gymnodimine and its analogues (gymnodimine acetate, gymnodimine methyl carbonate and gymnodamine) on cellular viability were tested using the Neuro2a neuroblastoma cell line. Concentrations of toxins up to 10µM had variable effects on reducing cell number as determined using the MTT assay and no effects on the expression of a number of signal transduction proteins (c-Jun, ATF-2, ATF-3) which are sensitive to cellular stress. However, pre-exposure of Neuro2a cells to 10µM concentrations of toxins for 24h greatly sensitized these cells to the apoptotic effects of another algal toxin, okadaic acid. These results suggest that gymnodimine and its analogues sensitize Neuro2a cells to cytotoxins and raise the possibility that algal blooms involving the production of both okadaic acid-type molecules and gymnodimine may generate greater cytotoxicity and pose a greater public health problem. Furthermore, our studies establish the Neuro2a cell line as a potentially high-throughput cellular system sensitive to the pharmacological effects of gymnodimine and analogues, and as a potential screen for algal-derived toxins.

Potent and long-acting dimeric inhibitors of influenza virus neuraminidase are effective at a once-weekly dosing regimen.

Dimeric derivatives (compounds 7 to 9) of the influenza virus neuraminidase inhibitor zanamivir (compound 2), which have linking groups of 14 to 18 atoms in length, are approximately 100-fold more potent inhibitors of influenza virus replication in vitro and in vivo than zanamivir. The observed optimum linker length of 18 to 22 A, together with observations that the dimers cause aggregation of isolated neuraminidase tetramers and whole virus, indicate that the dimers benefit from multivalent binding via intertetramer and intervirion linkages. The outstanding long-lasting protective activities shown by compounds 8 and 9 in mouse influenza infectivity experiments and the extremely long residence times observed in the lungs of rats suggest that a single low dose of a dimer would provide effective treatment and prophylaxis for influenza virus infections.

Gene-engineered T cells as a superior adjuvant therapy for metastatic cancer.

The major limiting factor in the successful application of adjuvant therapy for metastatic disease is the lack of adjuvant specificity that leads to severe side effects. Reasoning that T cells of the immune system are highly specific, we generated tumor-specific T cells by genetic modification of mouse primary T cells with a chimeric receptor reactive with the human breast cancer-associated Ag erbB-2. These T cells killed breast cancer cells and secreted IFN-gamma in an Ag-specific manner in vitro. We investigated their use against metastatic breast cancer in mice in an adjuvant setting, and compared their effectiveness with the commonly applied adjuvants doxorubicin, 5-fluorouracil, and herceptin. Mice were inoculated orthotopically with the human erbB-2-expressing spontaneously metastatic mouse breast cancer 4T1.2 in mammary tissue, and the primary tumor was surgically removed 8 days later. Significant metastatic disease was demonstrated in lung and liver at the time of surgery on day 8 with increased tumor burden at later time points. T cell adjuvant treatment of day 8 metastatic disease resulted in dramatic increases in survival of mice, and this survival was significantly greater than that afforded by either doxorubicin, 5-fluorouracil, or herceptin.

Highly potent and long-acting trimeric and tetrameric inhibitors of influenza virus neuraminidase.

A set of trimeric and tetrameric derivatives 6-11 of the influenza virus neuraminidase inhibitor zanamivir 1 have been synthesized by coupling a common monomeric zanamivir derivative 3 onto various multimeric carboxylic acid core groups. These discrete multimeric compounds are all significantly more antiviral than zanamivir and also show outstanding long-lasting protective activity when tested in mouse influenza infectivity experiments.