Diclofenac sodium adsorption on activated carbon: experimental, modeling and bayesian statistics.

The present study modeled the adsorption process of the drug diclofenac sodium on activated charcoal. For this purpose, a mass balance-based model was used considering a fixed bed column. The mass transfer rate in the solid phase was represented by a driving force model proposed in this study, and a gamma exponent with a range of 0 > γ ≤ 2 was assigned to the model. Different isotherms were adopted to represent the equilibrium at the solid/liquid interface: the Langmuir, Freundlich, Sips and Redlich-Peterson isotherms. The modeling was approached from the perspective of Bayesian statistics, and the Markov chain Monte Carlo method was used for parameter estimation. Model validation was performed with experimental data obtained under different operating conditions of initial concentration ($C_{0.

Pre- and perinatal exposures associated with molar incisor hypomineralization: Birth cohort, Brazil.

OBJECTIVE: To analyze prenatal and perinatal stressors associated with molar incisor hypomineralization (MIH) in adolescents. METHODS: Prospective cohort study collected prenatal (socioeconomic status, maternal age, number of prenatal visits, smoking, obesity during pregnancy, abortion history, gestational hypertension) and perinatal stressors (type of delivery, gestational age, birth weight, intensive care unit-ICU at birth). The outcome was MIH at 18-19 years follow-up (n = 590). MIH was defined according to the Ghanim criteria - Model I. We performed a sensitivity analysis, including opacities demarcated in index tooth, incisive or molars, Model II. Through structural equation modeling, we analyzed direct and mediating pathways between multiple stressors with outcomes. RESULTS: MIH was observed in 15.25% (n = 90), and opacities demarcated in any index tooth were observed in 22.8% of adolescents (n = 135). In Model I, no stressor explained MIH significantly, although we watched high standardized coefficients (SC) for low birth weight (SC = 0.223, p = 0.147), lower gestational age (SC = 0.351; p = 0.254), and ICU admission (SC = 0.447, p = 0.254). In Model II, advanced maternal age (SC = 0.148; p < 0.05) and not undergoing prenatal care (SC = 0.384, p < 0.03) explained opacities demarcated in incisors or molars. CONCLUSION: Advanced maternal age and not undergoing prenatal care were associated with MIH lesion-like in incisors or molars.

Leptin levels, seasonality and thermal acclimation in the Microbiotherid marsupial Dromiciops gliroides: Does photoperiod play a role?

Mammals of the Neotropics are characterized by a marked annual cycle of activity, which is accompanied by several physiological changes at the levels of the whole organism, organs and tissues. The physiological characterization of these cycles is important, as it gives insight on the mechanisms by which animals adjust adaptively to seasonality. Here we studied the seasonal changes in blood biochemical parameters in the relict South American marsupial Dromiciops gliroides ("monito del monte" or "little mountain monkey"), under semi-natural conditions. We manipulated thermal conditions in order to characterize the effects of temperature and season on a battery of biochemical parameters, body mass and adiposity. Our results indicate that monitos experience an annual cycle in body mass and adiposity (measured as leptin levels), reaching a maximum in winter and a minimum in summer. Blood biochemistry confirms that the nutritional condition of animals is reduced in summer instead of winter (as generally reported). This was coincident with a reduction of several biochemical parameters in summer, such as betahydroxybutyrate, cholesterol, total protein concentration and globulins. Monitos seem to initiate winter preparation during autumn and reach maximum body reserves in winter. Hibernation lasts until spring, at which time they use fat reserves and become reproductively active. Sexual maturation during summer would be the strongest energetic bottleneck, which explains the reductions in body mass and other parameters in this season. Overall, this study suggests that monitos anticipate the cold season by a complex interaction of photoperiodic and thermal cues.

The Effect of 3% Phosphate Ascorbyl Gel on Bond Strength of Composite Resin to Enamel treated with 35% Hydrogen Peroxide.

OBJECTIVE: To evaluate the effect of 3% phosphate ascorbyl gel (PA) in different times onto the microshear bond strength of composite resin (CR) to bovine enamel treated with 35% hydrogen peroxide (HP). MATERIALS AND METHODS: Thirty enamel blocks of bovine incisors were made and divided into 5 groups (n = 6) with three specimens per group (n = 18), according to treatment: G1= No bleaching + CR; G2 = HP + CR after 15d; G3 = HP + CR after 24 hours; G4 = HP + PA (15 min) + CR after 24 hours; G5 = HP + PA (2 hours) + CR after 24 hours. The resin cylinders were made by Tygon matrices. Microshear bond strength test was performed using universal testing machine with a 50N load at a speed of 0.5 mm/min. Fracture modes were assessed by a stereomicroscope 40 ×. Microshear bond strength values were submitted to the analysis of variance (ANOVA) one-way and Tukey test (p < 0.05). RESULTS: G1 had significant results when compared to G3 and G5 (p < 0.01). However, G2, G3, G4 and G5 have showed no significant differences among groups (p > 0.05). Failure modes were categorized into adhesive (90%) and mixed (10%). CONCLUSION: The use of 3% phosphate ascorbyl gel for 15 minutes was able to improve bond strength of composite resin to bleached bovine enamel, but when 3% phosphate ascorbyl gel was applied during 40 minutes it negatively interfered in the adhesion of the resin to bleached bovine enamel.

The effects of poly-unsaturated fatty acids on the physiology of hibernation in a South American marsupial, Dromiciops gliroides.

Many mammals hibernate, which is a profound lethargic state of several weeks or months during winter, that represents a transitory episode of hetherothermy. As with other cases of dormancy, the main benefit of hibernation seems to be energy saving. However, the depth and duration of torpor can be experimentally modified by the composition of food, especially by fattyacid composition. In eutherians, diets rich in unsaturated fatty acids (i.e., fatty acids with at least one double bond) lengthen torpor, reduce metabolism and permit hibernation at lower temperatures. Here we studied whether diets varying in fatty acid composition have an effect on the physiology of hibernation in a South American marsupial, Dromiciops gliroides. We designed a factorial experiment where thermal acclimation (two levels: natural versus constant temperature) was combined with diet acclimation: saturated (i.e., diets with high concentration of saturated fatty acids) versus unsaturated (i.e., diets with high concentration of unsaturated fatty acids). We measured energy metabolism in active and torpid individuals, as well as torpor duration, and a suite of 12 blood biochemical parameters. After a cafeteria test, we found that D. gliroides did not show any preference for a given diet. Also, we did not find effects of diet on body temperature during torpor, or its duration. However, saturated diets, combined with high temperatures provoked a disproportionate increase in fat utilization, leading to body mass reduction. Those animals were more active, and metabolized more fats than those fed with a high proportion of unsaturated fatty acids (="unsaturated diets"). These results contrast with previous studies, which showed a significant effect of fatty acid composition of diets on food preferences and torpor patterns in mammals.

Thermoregulatory capacities and torpor in the South American marsupial, Dromiciops gliroides.

During periods of adverse conditions small endotherms depend on a continuous supply of food and energy to maintain body temperature. Thus, rapid and reversible phenotypic modifications at different organizational levels are key for an efficient use of resources and survival. In this study, we provide a quantitative description of thermoregulatory capacities and energy-saving strategies in the Chilean marsupial Dromiciops gliroides. In particular, we evaluated the effect of thermal acclimation on basal metabolic rate (BMR), thermal conductance (C) and torpor patterns, as well as the presence of non-shivering thermogenesis (NST) as a rewarming mechanism in this marsupial. Non-significant effects of thermal acclimation were observed in BMR, C and body mass, but cold-acclimated individuals exhibited significantly longer torpor bouts. Also, minimum body temperature during torpor, inter-bout body temperature and arousal rewarming rate were lower in cold-acclimated animals. Furthermore, we found that D. gliroides did not display NST in response to Norepinephrine. Hence, despite the high regulation of torpor of other species, D. gliroides shows low flexibility in the ability to adjust energy expenditure and insulation properties, and (as in other marsupials) NST do not seems to be important as thermoregulatory mechanism.

Comparative Study on the Periodontal Parameters Used in Diagnosing Periodontitis in Puerperae and Periodontitis' Relationship with the Birth of Preterm Infants: A Case-Control Study.

To compare different criteria for the diagnosis of periodontitis and to evaluate the association of this condition with prematurity, this case-control study was conducted on 283 mothers of infants, divided into two groups based on gestational age (cases: <37 weeks, controls: ≥37 weeks), with 71 cases and 212 controls. The periodontal evaluation included probing depth (PD), clinical attachment level (CAL), plaque index, and bleeding on probing (BOP). Participants were classified regarding periodontitis per 14 criteria based on different periodontal parameters. The criterion selected as the gold standard was the presence of at least four teeth with one or more sites with a PD ≥ 4 mm, CAL ≥ 3 mm, and BOP at the same site. The prevalence of periodontal disease ranged from 8.1% to 55.1%. Moreover, compared to the gold standard, the sensitivities of the other criteria were 100%, while specificity ranged from 50.4% to 96.4%. Periodontitis, defined by six of the selected criteria, was associated with prematurity after multivariate adjustment, with OR ranging from 1.85 to 2.69 and 95% CI from 1.01 to 5.56; one of them was the gold standard mentioned above. Measurements using the clinical parameters of PD, CAL, and bleeding at the same site (criteria 5, 6, 7, 8), CPI (criterion 10), and at least four teeth with a PD ≥ 4 mm and CAL ≥ 3 mm (criterion 11) to define periodontitis showed a statistically significant association (p < 0.05). Given this study's limitations, we can conclude that the diagnostic criteria for a periodontitis definition using a PD ≥ 4 mm and CAL ≥ 3 mm in two or more teeth, with BOP at the same site, seem stronger when detecting an association between periodontitis and prematurity.

Pericytes promote endothelial cell survival through induction of autocrine VEGF-A signaling and Bcl-w expression.

Endothelial cells (ECs) in blood vessels under formation are stabilized by the recruitment of pericytes, both in normal tissues and during angiogenesis in pathologic situations, including neoplasia. In the tumor vasculature, besides supporting the functionality of blood flow, pericytes protect ECs from antiangiogenic therapies, and have thus been implicated in clinical resistance to vascular targeting drugs. However, the molecular nature of the crosstalk between pericytes and ECs is largely unchartered. Herein, we identified pericyte-induced survival signals in ECs by isolation of vascular fragments derived from tumors that had been genetically or pharmacologically engineered to be either pericyte-rich or pericyte-poor. Pericytes induced the antiapoptotic protein Bcl-w in tumor ECs both in vivo and in vitro, thereby conveying protection from cytotoxic damage. The pericyte-dependent survival signaling in ECs was consequential to enforcement of an autocrine loop involving VEGF-A expression in ECs. Through molecular and functional studies, we delineated a signal transduction pathway in ECs downstream of integrin α(v) involving activation of NF-κB as the initiating event of the protective crosstalk from pericytes. Our elucidation of pericyte-derived pro-survival signaling in tumor ECs has potentially important implications for clinical development of antiangiogenic drugs, and suggests new therapeutic targets for rational multitargeting of cancer.

Trophic interactions of the endangered Southern river otter (Lontra provocax) in a Chilean Ramsar wetland inferred from prey sampling, fecal analysis, and stable isotopes.

Non-invasive methodological approaches are highly recommended and commonly used to study the feeding ecology of elusive and threatened mammals. In this study, we use multiple lines of evidence to assess the feeding strategies of the endangered Southern river otter, by determining seasonal prey availability (electrofishing), analysis of undigested prey remains (spraints), and the use of stable isotopes (δ(15)N and δ(13)C) in otter spraints (n = 262) and prey in a wetland ecosystem of southern Chile (39°49'S, 73°15'W). Fecal and isotopic analyses suggest that the otter diet is restricted to a few prey items, particularly the less-mobile, bottom-living, and larger prey such as crayfish (Samastacus spinifrons, 86.11%) and crabs (Aegla spp., 32.45%), supplemented opportunistically by cyprinids (Cyprinus carpio, 9.55%) and catfish (Diplomystes camposensis, 5.66%). The results suggest that the river otter is highly specialized in bottom foraging. Isotopic signatures of food sources and feces revealed a mid-upper trophic position for the Southern river otter, with either higher or lower δ(15)N values than their potential prey items. δ(13)C values for river otters were less enriched than their potential food resources. We suggest that due to their narrow trophic niche and possible dependence on only a few food items, this species may be highly vulnerable to the reduction in its prey populations. Finally, maintaining the ecological interactions between Southern river otters and their prey is considered a central priority for the survival of this endangered carnivore mammal.

Profound changes in blood parameters during torpor in a South American marsupial.

Seasonal torpor or hibernation is a phenomenon characterized by a physiological transition to dormancy (torpor) during challenging periods in terms of energy availability or metabolic load. Extensive physiological reprogramming and changes in gene-expression, immune function, oxygen transport and intermediate metabolism, occur during eutherian hibernation. Here we studied the seasonality of blood parameters, and during daily torpor, in a South American marsupial (Dromiciops gliroides). Seasonal trends in blood parameters showed an increase in hematological parameters during winter, and increases in total proteins, albumin and globulin during autumn. In contrast, torpor induced a drastic drop during most blood parameters. PCV dropped significantly 60%, as well as RBC (58%), hemoglobin concentration (58%), WBC (79%), including neutrophils (51%), eosinophils (84%) and lymphocytes (82%). Biochemical parameters also showed reductions: triglycerides (81%), proteins (32%), albumin (24%), globulins (38%), albumin (24%), creatinine (48%) and glucose (42%). Our results confirm some patterns observed in hibernating eutherians, such as leukopenia, probably caused by sequestration of white blood cells in organs. However, red blood cells and hemoglobin concentration also were reduced, which is to the best of our knowledge has not been reported for marsupials. The observed reduction in biochemical parameters suggests that marsupials, as in eutherians, change from carbohydrate-based to lipid-based metabolism during hibernation. However, the absence of increases in beta-hydroxybutyrate is puzzling. Finally, we found an increase (although non-significant after statistical correction for multiple comparisons) of creatine kinase which together with an increase in neutrophil/lymphocyte ratio could be indicative of muscle lysis and inflammation. These results indicate profound changes in standard physiological processes during torpor.

Ecological consistency across space: a synthesis of the ecological aspects of Dromiciops gliroides in Argentina and Chile.

Dromiciops gliroides is an arboreal marsupial found in the temperate forests of South America (36-43 °S). This species is the sole extant representative of the order Microbiotheria, and is a key seed disperser of many native plant species, including the keystone mistletoe Tristerix corymbosus. Here, we synthesized the current knowledge on the ecological aspects of this species, and compared the available information from Argentina and Chile. Population density (23 ± 2 (mean ± SE) individual/ha) and home range (1.6 ± 0.6 ha) appear to be relatively similar across a marked ecological gradient in the mainland, but lower densities (7 ± 2 individual/ha) and smaller home ranges (0.26 ± 0.04 ha) were detected at island sites. We detected regional variation in body condition in Chile, but there were no significant differences across a wider E-W gradient. Movement patterns fit a random walk model; such behavior might have important consequences in shaping plant's spatial patterns. Although our data suggest that D. gliroides is more tolerant to habitat disturbance than previously thought, its incapability to disperse across non-forested areas suggests that the rapid rate of habitat loss and fragmentation that characterizes southern temperate forests likely poses a serious threat to this species. These ecological similarities are surprising given that forests studied receive dramatically different rainfall and correspond to distinct forest types. The evidence synthetized here dispels some of the myths about this species but also stresses the need for more comprehensive ecological studies across its distribution range.

MYCMI-7: A Small MYC-Binding Compound that Inhibits MYC: MAX Interaction and Tumor Growth in a MYC-Dependent Manner.

Deregulated expression of MYC family oncogenes occurs frequently in human cancer and is often associated with aggressive disease and poor prognosis. While MYC is a highly warranted target, it has been considered "undruggable," and no specific anti-MYC drugs are available in the clinic. We recently identified molecules named MYCMIs that inhibit the interaction between MYC and its essential partner MAX. Here we show that one of these molecules, MYCMI-7, efficiently and selectively inhibits MYC:MAX and MYCN:MAX interactions in cells, binds directly to recombinant MYC, and reduces MYC-driven transcription. In addition, MYCMI-7 induces degradation of MYC and MYCN proteins. MYCMI-7 potently induces growth arrest/apoptosis in tumor cells in a MYC/MYCN-dependent manner and downregulates the MYC pathway on a global level as determined by RNA sequencing. Sensitivity to MYCMI-7 correlates with MYC expression in a panel of 60 tumor cell lines and MYCMI-7 shows high efficacy toward a collection of patient-derived primary glioblastoma and acute myeloid leukemia (AML) ex vivo cultures. Importantly, a variety of normal cells become G1 arrested without signs of apoptosis upon MYCMI-7 treatment. Finally, in mouse tumor models of MYC-driven AML, breast cancer, and MYCN-amplified neuroblastoma, treatment with MYCMI-7 downregulates MYC/MYCN, inhibits tumor growth, and prolongs survival through apoptosis with few side effects. In conclusion, MYCMI-7 is a potent and selective MYC inhibitor that is highly relevant for the development into clinically useful drugs for the treatment of MYC-driven cancer. Significance: Our findings demonstrate that the small-molecule MYCMI-7 binds MYC and inhibits interaction between MYC and MAX, thereby hampering MYC-driven tumor cell growth in culture and in vivo while sparing normal cells.

Bioenergetics and intestinal phenotypic flexibility in the microbiotherid marsupial (Dromiciops gliroides) from the temperate forest in South America.

The microbiotherid marsupial Dromiciops gliroides inhabits the temperate forests of the Southern hemisphere, facing seasonal nutritional and energetic bottlenecks due to its apparently facultative insectivory/frugivory. In order to understand the physiological processes behind this ecological pattern, we studied the morpho-physiological changes that D. gliroides exhibits after dietary acclimation, in a sample of 21 wild-caught individuals fed over 1 month with ad libitum diet of: (1) fruit, (2) insects or (3) a mix of insects and fruit. In addition, we measured oxygen consumption (VO(2)) at resting conditions. We also performed enzyme assays (sucrase, maltase, trehalase and aminopeptidase N) and measurements of organ morphology. We found that D. gliroides cannot fulfil its nutrient requirements only from insects or fruit. It needs a mixed diet in order to maintain its body mass and energy balance. However, as a response of diet acclimation, individuals showed several-fold changes in the activities of aminopeptidase-N, maltase and sucrase (but not trehalase). This result, both the magnitude of change and the simultaneous effects on three enzymes suggests that D. gliroides could exhibit adaptive phenotypic plasticity in the activity of intestinal enzymes. This study suggests also that D. gliroides, the only living representative of the Microbiotheria order, exhibits physiological adaptations to a generalist diet.

PTEN and DNA-PK determine sensitivity and recovery in response to WEE1 inhibition in human breast cancer.

Inhibition of WEE1 kinase by AZD1775 has shown promising results in clinical cancer trials, but markers predicting AZD1775 response are lacking. Here we analysed AZD1775 response in a panel of human breast cancer (BC) cell lines by global proteome/transcriptome profiling and identified two groups of basal-like BC (BLBCs): 'PTEN low' BLBCs were highly sensitive to AZD1775 and failed to recover following removal of AZD1775, while 'PTEN high' BLBCs recovered. AZD1775 induced phosphorylation of DNA-PK, protecting cells from replication-associated DNA damage and promoting cellular recovery. Deletion of DNA-PK or PTEN, or inhibition of DNA-PK sensitized recovering BLBCs to AZD1775 by abrogating replication arrest, allowing replication despite DNA damage. This was linked to reduced CHK1 activation, increased cyclin E levels and apoptosis. In conclusion, we identified PTEN and DNA-PK as essential regulators of replication checkpoint arrest in response to AZD1775 and defined PTEN as a promising biomarker for efficient WEE1 cancer therapy.

[Approaching the concept of mental health for indigenous peoples in Colombia]. / Aproximación a la concepción de la salud mental para los pueblos indígenas de Colombia.

This study sought to perceive the concept of mental health from the perspective of Colombian indigenous peoples. A qualitative survey from a historic-hermeneutical standpoint was conducted by means of interviews with a stratified intentional sample of 10 leaders with experience in indigenous mental health. After being duly transcribed, interview materials were analyzed using a grounded theory approach. Findings were structured around four analytical categories: mental health perspectives, elements, scenarios, and challenges. It was revealed that some indigenous people do not perceive mental health as an indigenous concept. Some perceive it from a morbicentric western perspective and others from an ancestral holistic standpoint. They describe positive elements (good living, spirituality, harmony with mother earth) and negative aspects (acculturation, discrimination, violence, disobedience). Mental health scenarios included the land itself and the health care system, in particular the Intercultural Indigenous Health System (SISPI). Challenges include the need to conduct more research on this topic, consolidating SISPI, and fostering ancestral knowledge.

Este estudio tuvo como objetivo comprender el concepto de salud mental desde la perspectiva de los pueblos indígenas de Colombia. Se realizó una investigación con enfoque cualitativo, de tipo histórico hermenéutico a través de entrevistas a una muestra intencional estratificada de 10 líderes con experiencia en el tema de salud indígena. Luego de transcritas, se realizó el análisis de las entrevistas con herramientas de la teoría fundada. Los hallazgos se centraron en cuatro categorías analíticas: perspectivas de la salud mental, elementos, escenarios y retos. Se encontró que algunos indígenas no asumen la salud mental como un concepto propio. Entre los restantes, algunos lo asumen desde una perspectiva occidental morbicéntrica y los demás desde una perspectiva ancestral holística. Se describen elementos positivos (buen vivir, espiritualidad, armonía con la madre tierra) y negativos (aculturación, discriminación, violencia, desobediencia). Los escenarios de la salud mental que mencionaron los participantes son el territorio y el sistema de salud, en particular el Sistema Indígena de Salud Propio e Intercultural- SISPI. Los retos incluyen la necesidad de generar más investigaciones en este tema, el avance en la consolidación del SISPI y el fortalecimiento de los saberes ancestrales.

Prediction of response to anti-cancer drugs becomes robust via network integration of molecular data.

Despite the widening range of high-throughput platforms and exponential growth of generated data volume, the validation of biomarkers discovered from large-scale data remains a challenging field. In order to tackle cancer heterogeneity and comply with the data dimensionality, a number of network and pathway approaches were invented but rarely systematically applied to this task. We propose a new method, called NEAmarker, for finding sensitive and robust biomarkers at the pathway level. scores from network enrichment analysis transform the original space of altered genes into a lower-dimensional space of pathways. These dimensions are then correlated with phenotype variables. The method was first tested using in vitro data from three anti-cancer drug screens and then on clinical data of The Cancer Genome Atlas. It proved superior to the single-gene and alternative enrichment analyses in terms of (1) universal applicability to different data types with a possibility of cross-platform integration, (2) consistency of the discovered correlates between independent drug screens, and (3) ability to explain differential survival of treated patients. Our new screen of anti-cancer compounds validated the performance of multivariate models of drug sensitivity. The previously proposed methods of enrichment analysis could achieve comparable levels of performance in certain tests. However, only our method could discover predictors of both in vitro response and patient survival given administration of the same drug.

Low-dose metronomic daily cyclophosphamide and weekly tirapazamine: a well-tolerated combination regimen with enhanced efficacy that exploits tumor hypoxia.

The recent clinical successes of antiangiogenic drug-based therapies have also served to highlight the problem of acquired resistance because, similar to other types of cancer therapy, tumors that initially respond eventually stop doing so. Consequently, strategies designed to delay resistance or treat resistant subpopulations when they arise have assumed considerable importance. This requires a better understanding of the various possible mechanisms for resistance. In this regard, reduced oxygenation is thought to be a key mediator of the antitumor effects of antiangiogenic therapies; accordingly, increased hypoxia tolerance of the tumor cells presents a potential mechanism of resistance. However, hypoxia can also be exploited therapeutically through the use of hypoxic cell cytotoxins, such as tirapazamine. With this in mind, we measured the oxygenation of PC-3 human prostate cancer xenografts subjected to chronic low-dose metronomic (LDM) antiangiogenic chemotherapy using cyclophosphamide given through the drinking water. We found that LDM cyclophosphamide impairs the oxygenation of PC-3 xenografts even during relapse, coinciding with reduced microvessel density. Combination of LDM cyclophosphamide with tirapazamine results in significantly improved tumor control in the PC-3, HT-29 colon adenocarcinoma, and MDA-MB-231 breast cancer human xenograft models without having a negative effect on the favorable toxicity profile of LDM cyclophosphamide. These results provide further evidence that reduced vascular dependence/increased hypoxia tolerance may be a basis for eventual resistance of tumors exposed to long-term LDM chemotherapy.

Targeted anti-vascular endothelial growth factor receptor-2 therapy leads to short-term and long-term impairment of vascular function and increase in tumor hypoxia.

Because antiangiogenic therapies inhibit the growth of new tumor-associated blood vessels, as well as prune newly formed vasculature, they would be expected to reduce the supply of oxygen and thus increase tumor hypoxia. However, it is not clear if antiangiogenic treatments lead only to consistent and sustained increases in hypoxia, or transient decreases in tumor hypoxia along with periods of increased hypoxia. We undertook a detailed analysis of an orthotopically transplanted human breast carcinoma (MDA-MB-231) over a 3-week treatment period using DC101, an anti-vascular endothelial growth factor receptor 2 antibody. We observed consistent reductions in microvascular density, blood flow (measured by high-frequency micro-ultrasound), and perfusion. These effects resulted in an increase in the hypoxic tumor fraction, measured with an exogenous marker, pimonidazole, concurrent with an elevation in hypoxia-inducible factor-1alpha expression, an endogenous marker. The increase in tumor hypoxia was evident within 5 days and remained so throughout the entire course of treatment. Vascular perfusion and flow were impaired at days 2, 5, 7, 8, 14, and 21 after the first injection, but not at 4 hours. A modest increase in the vessel maturation index was detected after the 3-week treatment period, but this was not accompanied by an improvement in vascular function. These results suggest that sustained hypoxia and impairment of vascular function can be two consistent consequences of antiangiogenic drug treatment. The implications of the results are discussed, particularly with respect to how they relate to different theories for the counterintuitive chemosensitizing effects of antiangiogenic drugs, even when hypoxia is increased.

A selective high affinity MYC-binding compound inhibits MYC:MAX interaction and MYC-dependent tumor cell proliferation.

MYC is a key player in tumor development, but unfortunately no specific MYC-targeting drugs are clinically available. MYC is strictly dependent on heterodimerization with MAX for transcription activation. Aiming at targeting this interaction, we identified MYCMI-6 in a cell-based protein interaction screen for small inhibitory molecules. MYCMI-6 exhibits strong selective inhibition of MYC:MAX interaction in cells and in vitro at single-digit micromolar concentrations, as validated by split Gaussia luciferase, in situ proximity ligation, microscale thermophoresis and surface plasmon resonance (SPR) assays. Further, MYCMI-6 blocks MYC-driven transcription and binds selectively to the MYC bHLHZip domain with a KD of 1.6 ± 0.5 µM as demonstrated by SPR. MYCMI-6 inhibits tumor cell growth in a MYC-dependent manner with IC50 concentrations as low as 0.5 µM, while sparing normal cells. The response to MYCMI-6 correlates with MYC expression based on data from 60 human tumor cell lines and is abrogated by MYC depletion. Further, it inhibits MYC:MAX interaction, reduces proliferation and induces massive apoptosis in tumor tissue from a MYC-driven xenograft tumor model without severe side effects. Since MYCMI-6 does not affect MYC expression, it is a unique molecular tool to specifically target MYC:MAX pharmacologically and it has good potential for drug development.

Publisher Correction: A DHODH inhibitor increases p53 synthesis and enhances tumor cell killing by p53 degradation blockage.

The original PDF version of this Article listed the authors as "Marcus J.G.W. Ladds," where it should have read "Marcus J. G. W. Ladds, Ingeborg M. M. van Leeuwen, Catherine J. Drummond et al.#".Also in the PDF version, it was incorrectly stated that "Correspondence and requests for materials should be addressed to S. Lín.", instead of the correct "Correspondence and requests for materials should be addressed to S. Laín."This has been corrected in the PDF version of the Article. The HTML version was correct from the time of publication.