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.

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.

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.

Maternal and fetal parameters in pregnant woman undergoing tocolysis with nifedipine / Parâmetros maternos e fetais em gestante submetida a tocólise com nifedipina

Abstract Objectives: to evaluate the effects of nifedipine with tocolysis under maternal and fetal parameters. Methods: a cohort study with 40 pregnant women admitted at a high-risk pregnancy ward to inhibit premature labor between September/2010 to May/2012. Nifedipine was used as a 20mg sublingual attack dose and maintained 20mg every six and eight hours orally. The variables of the analysis were fetal heart rate (FHR), maternal heart rate (MHR), systolic blood pressure (SBP) and diastolic blood pressure (DBP), and amniotic fluid index (AFI). All the variables were evaluated prior to administrating nifedipine and approximately after 6 hours and every 24 hours, until hospital discharge. Results: there were no modification of the FHR (p=0.48) and the SBP (p=0.29). The MHR increased after 24 hours, but with no statistical difference (p=0.08), returning to similar levels as at admission within 48 hours. The DBP decreased at 6 (p=0.04) to 72 hours, being stable afterwards. The AFI decreased significantly at 24, 48 and 72 hours. Conclusions: the use of high doses of nifedipine with tocolysis causes a decrease of the maternal's diastolic blood pressure and consequently decreases the amniotic fluid index, but probably without any clinical repercussions.

Resumo Objetivos: avaliar os efeitos da nifedipina utilizada na tocólise sobre os parâmetros maternos e fetais. Métodos: estudo de coorte incluindo 40 gestantes admitidas na enfermaria de alto risco para inibição do trabalho de parto prematuro entre setembro/2010 a maio/2012. Utilizou-se a nifedipina sublingual na dose de ataque de 20mg e uma manutenção de 20mg por via oral a cada seis e oito horas. As variáveis avaliadas foram os batimentos cardio-fetais (BCF), frequência cardíaca materna (FCM), pressão arterial sistólica (PAS) e diastólica (PAD) e índice de líquido amniótico (ILA). Todas as variáveis foram avaliadas antes da administração da nifedipina e aproximadamente após 6h e cada 24h até alta hospitalar. Resultados: não houve modificação dos BCF (p=0,48) e da PAS (p=0,29). A FCM aumentou após 24h, mas sem significância estatística (p=0,08) retornando a níveis similares ao da admissão com 48h. A PAD diminuiua partir de 6h (p = 0,04)até 72h, mantendo-se constante. O ILA diminuiu significativamente em 24h, 48h e 72h. Conclusão: a utilização de altas doses de nifedipina para tocóliseocasio na diminuição dos níveis pressóricos diastólicos maternos e consequentemente diminuição do ILA, mas provavelmente sem repercussões clínicas.

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

Resumen 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.

Abstract 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.

Índice de manejo quirúrgico en pacientes con cáncer ginecológico durante la pandemia por Covid-19: propuesta para instituciones y profesionales en ginecología oncológica / Index of surgical management in gynecological cancer patients during the covid-19 pandemic: a proposal for patients during the covid-19 pandemic: a proposal for institutions and gynecological onchology professionals institutions and gynecological onchology professional

El mundo vive una pandemia por un Coronavirus, llamado SARS-COV-2, que produce la COVID-19 (acrónimo del ingles coronavirus disease 2019), la cual ha generado un colapso en los sistemas de salud, haciendo que el manejo de otras enfermedades se convierta en un reto. De igual forma para los grupos oncológicos, la presencia de esta enfermedad, genera muchas dudas en la aplicación de los tratamientos estándares, los cuales se deben realizar lo antes posible, con el fin de ofrecer mejores resultados oncológicos. Se propone la creación de un índice ( COVID-19 Cáncer Index) , teniendo en cuenta variables clínicas, epidemiológicas y la disponibilidad de los recursos hospitalarios, útil para la toma de decisiones y el establecimiento del mejor tratamiento para una paciente con confirmación o alta sospecha de neoplasia ginecológica.

The world is experiencing a coronavirus pandemic called SARS-COV-2 which causes coronavirus disease 2019 (COVID 19). This has led to a collapse in health systems, making the management of other diseases a challenge. Similarly, the presence of this disease generates many doubts for oncological groups regarding the provision of standard treatments, which should be carried out as soon as possible, in order to ensure better oncological outcomes. We propose the creation of an index (COVID 19 Cancer Index) taking into account clinical and epidemiological variables and the availability of hospital resources, which are useful for decision making and determining the best treatment for a patient with confirmed or strongly suspected gynecological neoplasia.

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.

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.

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

The development of non-genotoxic therapies that activate wild-type p53 in tumors is of great interest since the discovery of p53 as a tumor suppressor. Here we report the identification of over 100 small-molecules activating p53 in cells. We elucidate the mechanism of action of a chiral tetrahydroindazole (HZ00), and through target deconvolution, we deduce that its active enantiomer (R)-HZ00, inhibits dihydroorotate dehydrogenase (DHODH). The chiral specificity of HZ05, a more potent analog, is revealed by the crystal structure of the (R)-HZ05/DHODH complex. Twelve other DHODH inhibitor chemotypes are detailed among the p53 activators, which identifies DHODH as a frequent target for structurally diverse compounds. We observe that HZ compounds accumulate cancer cells in S-phase, increase p53 synthesis, and synergize with an inhibitor of p53 degradation to reduce tumor growth in vivo. We, therefore, propose a strategy to promote cancer cell killing by p53 instead of its reversible cell cycle arresting effect.

Efecto de la temperatura de almacenamiento sobre la viabilidad de bacterias simbióticas fijadoras de nitrógeno utilizadas en la elaboración de inoculantes biológicos para arveja (Pisum sativum) y soya (Glycine max) / Effect of storage temperature on the viability of symbiotic nitrogen fixing bacteria used in the preparation of biological inoculants for crop pea (Pisum sativum) and soybean (Glycine max)

El nitrógeno es un elemento requerido en grandes cantidades por los cultivos, sin embargo, en el suelo es uno de los elementos más limitantes ya que se encuentra en formas poco disponibles para las plantas y debe ser incorporado a través de la fertilización química o por medio de la fijación biológica llevada a cabo por microorganismos. Las bacterias simbióticas fijadoras de nitrógeno han sido ampliamente utilizadas para la elaboración de inoculantes, constituyéndose en una alternativa viable para mejorar el rendimiento de los cultivos a través de un mejor suministro de este elemento. Estos inoculantes para que puedan ser comercializados deben cumplir con ciertos requisitos de calidad que garanticen el buen funcionamiento del producto. En este estudio se evalúo la viabilidad de las cepas ICA L9 e ICA J96 en inoculantes almacenados a temperaturas de 4±2, 18±3 y 28±2 °C durante 180 días y la actividad biológica en plantas de arveja y soya. Las cepas utilizadas pertenecen al Banco de Germoplasma de Microorganismos de Corpoica; para los ensayos biológicos se emplearon semillas de arveja variedad "Santa Isabel" y soya variedad "Corpoica Superior 6". Al evaluar la viabilidad de los inoculantes almacenados a 4±2 °C, 18±3 °C y 28±2 °C, se evidenció que la temperatura no afectó la supervivencia y la concentración de rizobios después de 180 días de almacenamiento permitiendo observar un número de unidades formadoras de colonias por gramo superior a 108, valor que garantiza la calidad del inoculante. En cuanto a la actividad biológica, se observó que las cepas inoculadas fueron infectivas y efectivas para la fijación biológica del nitrógeno, compa­rado con los testigos absolutos.

Nitrogen is an element required in large amounts by most crops, however, in soil is one of the most limiting and located in ways not available to the plant and must be incorporated through chemical fertilization or by biological fixation conducted by microorganisms. Fixing symbiotic bacteria nitrogen has been widely used for the production of inoculants, becoming a viable alternative to improve crop yields through a better supply of this element. These inoculants to be marketed must meet certain requirements quality to ensure the smooth operation of the product. This study assessed the viability of the strains ICA L9 and ICA J96 inoculants stored at temperatures 4±2, 18±3 y 28±2°C for 180 days and the biological activity in pea and soybean plants. The strains used belong to the collection of work Germplasm Bank CORPOICA Microorganisms; for biological assays were used pea seeds variety "Santa Isabel" and soybean variety "Superior Corpoica 6". In assessing the viability of inoculants stored at 4±2°C, 18±3°C y 28±2°C, evidenced that the temperature did not affect the survival and the concentration of rhizobia after 180 days of storage allowing to observe a number of colony forming units per gram than 10s, value that guarantees the quality of the inoculant. Concerning the biological activity, it was observed that the strains were inoculated infective and effective biological nitrogen fixation, absolute compared to controls.

Photocatalytic degradation of nicotine in an aqueous solution using unconventional supported catalysts and commercial ZnO/TiO2 under ultraviolet radiation.

Nicotine, a highly toxic alkaloid, has been detected in effluents, surface and groundwater and even bottled mineral water. The present work studied the photocatalytic degradation of nicotine in aqueous solution, under ultraviolet irradiation. The experiments were carried out using commercial (ZnO, TiO2) and non-conventional catalysts, which were prepared from industrial and laboratory waste. Two experimental designs (CCD) were performed for both commercial catalysts, and initial nicotine concentration, catalyst concentration and initial solution pH effects were studied. Then, the synthesized catalysts were tested under the optimal conditions which were found through CCDs. Using commercial catalysts, about 98% of the alkaloid was degraded by ZnO, and 88% by TiO2, in 1h. Among the non-conventional catalysts, the highest photocatalytic degradation (44%) was achieved using the catalyst prepared from a petrochemical industry residue.

Deficiency for endoglin in tumor vasculature weakens the endothelial barrier to metastatic dissemination.

Therapy-induced resistance remains a significant hurdle to achieve long-lasting responses and cures in cancer patients. We investigated the long-term consequences of genetically impaired angiogenesis by engineering multiple tumor models deprived of endoglin, a co-receptor for TGF-ß in endothelial cells actively engaged in angiogenesis. Tumors from endoglin-deficient mice adapted to the weakened angiogenic response, and refractoriness to diminished endoglin signaling was accompanied by increased metastatic capability. Mechanistic studies in multiple mouse models of cancer revealed that deficiency for endoglin resulted in a tumor vasculature that displayed hallmarks of endothelial-to-mesenchymal transition, a process of previously unknown significance in cancer biology, but shown by us to be associated with a reduced capacity of the vasculature to avert tumor cell intra- and extravasation. Nevertheless, tumors deprived of endoglin exhibited a delayed onset of resistance to anti-VEGF (vascular endothelial growth factor) agents, illustrating the therapeutic utility of combinatorial targeting of multiple angiogenic pathways for the treatment of cancer.

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.

Reparo dos tecidos periodontais após cirurgia ressectiva: relato de caso clínico / Repair of periodontal tissues following ressective surgery: a case report

The aim of this study was to demonstrate the reestablishment of the biological width with periodontal surgery and follow up the positional changes of the gingival and bone tissues after a six-month healing period. The measures of gingival margin, level of relative attachment, mucogingival junction, cervical preparation cavity were obtained before full-thickness flap with osteotomy and osteoplasty, and repeated in the postoperative. Radiographs were made to verify the formation of lamina dura at that period of evaluation. The results have showed: 1) mucogingival junction without major changes after 6 months, 2) mean of level of relative attachment 0,5 mm; 3) coronary migration of gingival margin at the 2-month and stabilization at the 4-month periods; 4) stabilization of the papilla according to the shape and to the fold after 6 months and to the appearance in 4 months, 5) visible lamina dura was observed at proximal alveolar crests at the 6-month period. It can conclude that there was repair of gingival and bone tissues, thus enabling the placement of accurate prosthetic procedure in order to provide aesthetics, function, and the maintenance of periodontal health.

Hardening of the national flower of Colombia, the threatened Cattleya trianae (Orchidaceae), from in vitro culture with previous invigoration phase

Cattleya trianae is an endemic species from the tropical rainforest in the Colombian Andes. Its survival is currently threatened due to habitat loss and commercial overexploitation. This study evaluates ten substrates, some organic (pine bark, coconut fiber and wood shavings), some inert icopor (polystyrene foam), vegetable coal and their combinations, and the effects these have on morphometric and phenotypic traits in the hardening phase of 250 plants of C. trianae cultivated in vitro. Recorded data include percent survival, length of longest leaf, biomass (wet weight) and number of roots and leaves at the beginning and at the end of the experiment. After the hardening phase, the plants were taken to a greenhouse and later to the natural environment. Coconut fiber alone or mixed in equal parts with pine bark and coal was the most efficient substrate when percent survival (80±SE=0.3742), biomass, and leaf length were evaluated. Hardened plants displayed qualitative characteristics such as vigor, hardness and waxy texture, strength of green coloration in the leaves, and velamen formation. Under greenhouse conditions, plants grew better with filtered light, relative humidity bordering on 80 %, permanent aeration, misting with water, and an average temperature of 25±2 °C. Invigorated plants were firmly anchored on their host trees. Rev. Biol. Trop. 55 (2): 681-691. Epub 2007 June, 29.

Cattleya trianae es una especie endémica de los bosques tropicales de los Andes colombianos. Actualmente se encuentra amenazada por la disminución de su hábitat natural y la sobreexplotación con fines comerciales. En este estudio se evaluó el efecto de diez tratamientos con sustratos biológicos (corteza de pino, fibra de coco y viruta) e inertes (esferitas de "icopor" y carbón vegetal) en diferentes combinaciones, sobre aspectos morfométricos y fenotípicos en la etapa de endurecimiento de 250 vitroplantas de C. trianae. Se registró porcentaje de supervivencia, longitud de la hoja, biomasa en peso fresco, número de raíces y hojas al inicio y al final del experimento. Al finalizar la fase de endurecimiento, las vitroplantas fueron llevadas a invernadero y posteriormente a ambiente natural. La fibra de coco sola ó mezclada en partes iguales con pino y carbón vegetal, fue el sustrato más eficiente cuando se evaluó el porcentaje de supervivencia (80 % ±SE=0.3742), biomasa en peso fresco y longitud de hoja. Las plantas endurecidas mostraron características cualitativas como vigorosidad, textura coriácea y cerosa, verdor intenso en sus hojas y velamen. En condiciones de invernadero las plantas se desarrollan mejor con luz filtrada, humedad relativa alrededor del 80 %, aireación continua, nebulización y temperatura promedio de 25±2 °C. Las plantas vigorizadas mostraron buen anclaje y adaptación en árboles.

Therapy-induced acute recruitment of circulating endothelial progenitor cells to tumors.

The contribution of bone marrow-derived circulating endothelial progenitor cells (CEPs) to tumor angiogenesis has been controversial, primarily because of their low numbers in blood vessels of untreated tumors. We show that treatment of tumor-bearing mice with vascular disrupting agents (VDAs) leads to an acute mobilization of CEPs, which home to the viable tumor rim that characteristically remains after such therapy. Disruption of this CEP spike by antiangiogenic drugs or by genetic manipulation resulted in marked reductions in tumor rim size and blood flow as well as enhanced VDA antitumor activity. These findings also provide a mechanistic rationale for the enhanced efficacy of VDAs when combined with antiangiogenic drugs.

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.

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.