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Nowadays, extracellular DNA or circulating cell-free DNA is considered to be a molecule with clinical applications (diagnosis, prognosis, monitoring of treatment responses, or patient follow-up) in diverse pathologies, especially in cancer. Nevertheless, because of its molecular characteristics, it can have many other functions. This review focuses on the participation of extracellular DNA (exDNA) in fundamental processes such as cell signaling, coagulation, immunity, evolution through horizontal transfer of genetic information, and adaptive response to inflammatory processes. A deeper understanding of its role in each of these processes will allow development of better tools to monitor and control pathologies, as well as helping to generate new therapeutic options, beyond the applicability of DNA in liquid biopsy.

Birds are important hosts for the development of the immature stages of several tick species that are vectors for disease-causing microorganisms in animals and humans. Colombia has the highest number of bird species worldwide; however, there is scarce data on the role of birds in the circulation of ticks and their associated pathogens, such as rickettsiae. The department of Arauca has a high diversity of resident and migratory (boreal and austral) birds and ticks associated with the transmission of Rickettsia. The objective of this research was to identify tick species parasitizing birds and to detect Rickettsia species in these ectoparasites. We conducted samplings in the municipalities of Arauca, Cravo Norte, and Tame between November of 2018 and August of 2019. Birds were captured using mist nets and examined for the presence of tick species. The collected ticks were morphologically and molecularly identified. Furthermore, we detected rickettsiae in ticks by amplifying fragments of the citrate synthase (gltA) and outer membrane protein (ompB) genes. We captured 606 birds belonging to 25 families and 115 species. Tick infestation rate was 3.3% (20/606) in the birds captured and eight new associations between wild birds and ticks are reported for the American continent. We identified four tick species: Amblyomma nodosum, Amblyomma longirostre, Amblyomma mixtum, and Amblyomma sp.. Moreover, we confirmed the presence of Rickettsia parkeri strain Atlantic rainforest in A. nodosum, a medically-relevant rickettsia due to cases of rickettsiosis in the American continent. This finding manifests the importance of wild birds as hosts and dispersal agents of ticks infected with pathogenic rickettsiae, as well as the need to monitor migratory birds in the Orinoquia and other regions of Colombia and America., Graphical abstract Image 1, Highlights • Wild birds are important hosts of ticks of the family Ixodidae that are infected by pathogen rickettsiae. • This study provides knowledge on the interactions between birds, ticks, and rickettsiae in Colombia. • We report new associations between wild birds and ticks in the American continent.

Pérez-Cárdenas E, Taja-Chayeb L, Trejo-Becerril C, et al. Onco Targets Ther. 2018;11:8823–8833. The authors have advised that an error during the preparation of the slide images for Figure 2C, on page 8827, led to the inadvertent duplication of features on the HV slide. All the original data was retained and the correct image for the HV treated chamber was used as a replacement. The correct Figure 2 is shown in Download Article. &nbsp

CD8+ T cell-mediated immune response plays a major role in the clearance of virus-infected cells, including human papillomavirus (HPV). The effective treatment of women with normal cytology but persistent high risk-HPV infection or with low-grade intraepithelial lesions could take advantage of novel strategies based on vaccination with viral immunological targets with a wide spectrum of cross-protection. The helicase E1, expressed early during viral replication in HPV infection, is among the most conserved papillomavirus proteins, which makes it a good vaccine candidate. In the present study, we examined E1-specific CD8+ T cell and NK immune responses in a mouse model with α-galactosylceramide (α-GalCer) as an adjuvant. We found that mice immunized with E1 combined with α-GalCer elicited an E1-specific CD8+ T and NK cell cytotoxic responses, which correlated with growth inhibition of grafted melanoma B16-F0 cells expressing E1, both in prophylactic and therapeutic protocols.

Enrique Pérez-Cárdenas,1 Lucía Taja-Chayeb,1 Catalina Trejo-Becerril,1 José Chanona-Vilchis,2 Alma Chávez-Blanco,1 Guadalupe Domínguez-Gómez,1 Elizabeth Langley,1 Alejandro García-Carrancá,3,4 Alfonso Dueñas-González3,4 1Division of Basic Research, Instituto Nacional de Cancerología, Mexico City, Mexico; 2Department of Pathology, Instituto Nacional de Cancerología, Mexico City, Mexico; 3Unit of Biomedical Research on Cancer, Biomedical Research Institute, Universidad Nacional Autonoma de Mexico, Mexico City, Mexico; 4Unit of Biomedical Research on Cancer, Instituto Nacional de Cancerologia, Mexico City, Mexico Introduction: Metastasis involves the accumulation of genetic and epigenetic alterations leading to activation of prometastatic genes and inactivation of antimetastatic genes. Among epigenetic alterations, DNA hypermethylation and histone hypoacetylation are the focus of intense translational research because their pharmacological inhibition has been shown to produce antineoplastic activity in a variety of experimental models. Aims: This study aimed to evaluate the antimetastatic effect of the DNA-methylation inhibitor, hydralazine, and the histone deacetylase inhibitor, valproic acid. Methods: NIH 3T3-Ras murine cells were treated with hydralazine and valproic acid to evaluate their effects upon cell proliferation, cell motility, chemotaxis, gelatinase activity, and gene expression. Lung metastases were developed by intravenous injection of NIH 3T3-Ras cells in BALB/c nu/nu mice and then treated with the drug combination. Results: Treatment induced a growth-inhibitory effect on NIH 3T3-Ras cells, showed a trend toward increased gelatinase activity of MMP2 and MMP9, and inhibited chemotaxis and cell motility. The combination led to a strong antimetastatic effect in lungs of nude mice. Conclusion: Hydralazine and valproic acid, two repositioned drugs as epigenetic agents, exhibit antimetastatic effects in vitro and in vivo and hold potential for cancer treatment. Keywords: hydralazine, valproic acid, epigenetics, metastasis, ras

It is estimated that up to 10% of gastric carcinomas show familial aggregation. In contrast, around 1-3 % (approximately 33,000 yearly) are genuinely hereditary. Hereditary diffuse gastric cancer (HDGC) is a rare malignancy characterized by autosomal dominant inheritance of pathological variants of the CDH1 and CTNNA1 genes encoding the adhesion molecules E-cadherin and α-catenin, respectively. The multifocal nature of the disease and the difficulty of visualizing precursor lesions by endoscopy underscore the need to be aware of this malignancy as surgical prevention can be fully protective. Here, we provide an overview of the main epidemiological, clinical, genetic, and pathological features of HDGC, as well as updated guidelines for its diagnosis, genetic testing, counseling, surveillance, and management. We conclude that HDGC is a rare, highly penetrant disease that is difficult to diagnose and manage, so it is necessary to correctly identify it to offer patients and their families’ adequate management following the recommendations of the IGCL. A critical point is identifying a mutation in HDGC families to determine whether unaffected relatives are at risk for cancer.

Los estudios sobre salud pública y biodiversidad de mamíferos silvestres incluyen un componente genético. Para las muestras de sangre, se debe tener condiciones óptimas de colección, ya que pueden afectar la preservación y la extracción del ADN. Este estudio evaluó el uso de métodos de preservación de ADN líquido y seco y métodos de extracción de ADN comerciales y no comerciales, en muestras de sangre, recolectadas en campo. Para ello, se recogieron 264 muestras de sangre totales de mamíferos salvajes. Se preservó un primer grupo de muestras en clorhidrato de guanidina (GuHCl) y se extrajo el ADN, utilizando seis kits comerciales: Bioline, Norgen, Invitrogen, Promega y Qiagen, además de dos protocolos no comerciales: fenol-cloroformo isoamil alcohol (PC) y guanidina tiocianato (GIT). Otro grupo de muestras, se preservó en tarjetas Whatman® FTA® y se extrajo el ADN, con PC y GIT. Las extracciones con GIT y PC mostraron los valores y variaciones más altas en la concentración de ADN (ng/µL), mientras que el kit comercial mostró una baja variación. La preservación de la muestra en tarjetas Whatman® FTA® proporcionó una baja variación y cantidad de ADN extraído, en comparación con el uso de GuHCl. En cuanto a la calidad del ADN, los kits comerciales produjeron una mayor pureza (A260/280), mientras que los protocolos basados en GIT y PC proporcionaron resultados muy variables. Además, el uso de GIT y PC originó una mayor cantidad de ADN, pero de calidad variable. En general, la extracción basada en kits comerciales y la conservación Whatman® FTA® permitió obtener calidades y cantidades de ADN más estandarizadas. Studies on public health and wild mammal biodiversity include a genetic component. For blood samples, there must be optimal sample collection conditions since these can affect DNA preservation and extraction. This study evaluated the use of liquid and dry DNA preservation methods and commercial and non-commercial DNA extraction methods on field-collected blood samples. For this, 264 total blood samples were collected from wild mammals. A first group of samples was preserved in guanidine hydrochloride (GuHCl) and DNA was extracted using six commercial kits: Bioline, Norgen, Invitrogen, Promega, and Qiagen, in addition to phenol-chloroform isoamyl alcohol (PC) and guanidine thiocyanate (GIT). Another group of samples was preserved in Whatman® FTA® cards and DNA was extracted with PC and GIT. The extractions with GIT and PC showed the highest values (ng/µL) and variation in DNA concentration, while the commercial kit showed low variation. Sample preservation in Whatman® FTA® cards provided low variation and quantity of the extracted DNA compared with the use of GuHCl. Concerning DNA quality, the commercial kits yielded higher purity, while GIT and PC-based protocols provided highly variable results. Furthermore, the use of GIT and PC yielded a higher amount of DNA, yet, of variable quality. Overall, extraction based on commercial kits and Whatman® FTA® preservation allowed obtaining more standardized DNA qualities and quantities Incluye referencias bibliográficas

Introducción: Los esfuerzos terapéuticos orientados a atender las micosis por Candida spp. se han enfocado en el empleo de azoles; sin embargo, en la literatura científica se discute su beneficio, por los amplios y descritos mecanismos de resistencia. Objetivo: Describir los mecanismos de resistencia al fluconazol expresados por la especie Candida glabrata, con la intención de que sean considerados dentro de las variables de elegibilidad para la intervención. Método: Se realizó una revisión integrativa utilizando la pregunta orientadora: ¿cuáles son los mecanismos de resistencia al fluconazol expresados por la especie Candida glabrata? Veintinueve estudios obtenidos de la base de datos PubMed cumplieron los criterios del análisis crítico propuesto por el instrumento PRISMA, utilizado para la selección de los artículos incluidos para su revisión en este manuscrito. Las categorías bajo las cuales se organizaron los elementos de análisis fueron: sobrexpresión de bombas de eflujo y modificaciones en la enzima lanosterol 14-alfa-desmetilasa. Resultados: Los mecanismos de resistencia al fluconazol expresados por Candida glabrata están determinados principalmente por la regulación a la alza de bombas de adenosina-trifosfato Binding Cassette (ABC) y por la modificación del punto de unión con su blanco farmacológico: la enzima lanosterol 14-alfa-desmetilasa. Conclusión: Los mecanismos de resistencia expresados por Candida glabrata se asocian con la modificación estructural de la diana farmacológica y la sobreexpresión de bombas de eflujo de manera diferencial a otras especies. Se sugiere que Candida glabrata es intrínsecamente menos susceptible al fluconazol.

Introduction: Rickettsioses are zoonotic diseases caused by pathogenic bacteria of the genus Rickettsia and transmitted to man by means of arthropod vectors such as ticks, fleas, mites and lice. Historically, Caldas Department has reported a significant number of cases of murine typhus to the Colombian national health surveillance system, and consequent studies of flea-borne rickettsiosis identified the circulation of Rickettsia typhi and Rickettsia felis in multiple municipalities. Our aim was to genotype species of Rickettsia detected in fleas collected from domestic and wild mammals in Caldas. Methodology: Flea samples were taken by convenience sampling from dogs, cats and wild mammals (rodents and marsupials) in 26 municipalities. Specimens were classified by current taxonomic keys and pooled for DNA extraction and molecular screening for Rickettsia spp. by PCR amplification of gltA, htrA and sca5 genes. Positive samples were genotyped by enzyme digestion (htrA) and sequencing. Results: A total of 1388 flea samples were collected. Rickettsia DNA was amplified in 818 (gltA), 883 (htrA) and 424 (sca5) flea pools. Alignment analysis with available Rickettsia DNA sequences showed greater similarity with R. asembonensis (gltA) and with R. felis (sca5 and htrA). Restriction pattern was compatible with R. felis. R. typhi was not identified. Conclusion: The present study confirms the presence and high prevalence of R. asembonensis and R. felis in fleas from domestic and wild animals in different municipalities from Caldas Department.

Cancer cells have increased glycolysis and glutaminolysis. Their third feature is increased de novo lipogenesis. As such, fatty acid (FA) synthesis enzymes are over-expressed in cancer and their depletion causes antitumor effects. As fatty acid synthase (FASN) plays a pivotal role in this process, it is an attractive target for cancer therapy.This is a review of the lipogenic phenotype of cancer and how this phenomenon can be exploited for cancer therapy using inhibitors of FASN, with particular emphasis on orlistat as a repurposing drug.Disease stabilization only has been observed with a highly selective FASN inhibitor used as a single agent in clinical trials. It is too early to say whether the absence of tumor responses other than stabilization results because even full inhibition of FASN is not enough to elicit antitumor responses. The FASN inhibitor orlistat is a 'dirty' drug with target-off actions upon at least seven targets with a proven role in tumor biology. The development of orlistat formulations suited for its intravenous administration is a step ahead to shed light on the concept that drug promiscuity can or not be a virtue.

The Colombian Tropical Andes are one of the regions with highest bird diversity on Earth. However, information on bird morphology, reproductive phenology, and molt is particularly scarce in this region. Also, this region is heavily impacted by deforestation, and it is vulnerable to climate change. Hence, providing baseline information on life history and morphological traits will be essential to support future research on functional diversity, climate change effects, conservation, evolution, and phenology. To fill this gap, we have compiled information on bird distribution, morphology, molt, and reproductive phenology at 52 localities of the Department of Caldas, covering an elevation range between 148 and 3845 m. This compilation comprises a wide range of habitats, including native forests, forestry plantations, croplands, and paramo. Our database presents information for 3,398 records belonging to 379 bird species (representing 23 orders, 53 families, and 258 genera). From those records, 2,843 correspond to information collected in the field between 2008 and 2019, and the remaining 555 records correspond to specimens deposited in the Natural History Museum of the Caldas University, collected between 1969 and 2014. We measured nine morphological traits from all specimens: total culmen, gape, bill width, bill depth, tarsus, wing length, tail length, total length, and mass. We also have reproductive condition information for 257 species and molt information available for 378 species. The information contained in this data set represents ~20% of the Colombian avifauna and ~11% of the bird species richness in South America. This data set is released for non-commercial use only. Credits should be given to this paper (i.e., proper citation), and the products generated with this database should be shared under the same license terms (CC BY-NC-SA).

Angiogenesis is the formation of new blood vessels from the pre-existing vasculature, it is essential for tumor growth and development. Quantitative evaluation of vascular parameters that allow to identify angiogenesis in early stages of tumor development could have an impact on lesion detection, cancer treatment, and patient outcome. The aim of this study was to define the factors related to contrast medium administration and image acquisition that allow to quantify vascular parameters from contrast-enhanced micro-computed tomography (CE-microCT) images. Type of contrast-medium, dose and image acquisition time were determined for single-energy (SE) and dual-energy (DE) CE-microCT imaging protocols using a tumor angiogenesis murine model. Enhancement and relative blood volume (rBV) were quantified from SE and DE images in muscle, tumor core, tumor periphery and the whole tumor. The differences in enhancement between muscle and the tumor regions quantified in SE images were not statistically significant. A statistically significant difference was found for rBV between muscle and tumor periphery in SE images, but the differences with tumor core and the whole tumor were not significant. For the DE protocol, more animals need to be included in the study in order to evaluate the differences in enhancement and rBV. Validation studies are also required to evaluate the correlation between rBV and angiogenesis biomarkers. Despite these limitations, rBV quantified from CE-microCT images seems to be a promising vascular parameter that could help to describe the angiogenic status of a tumor during cancer progression.

Several bacteria belonging to the genus Rickettsia are recognized as causal agents of diseases in domestic and wild animals and humans. These bacteria are considered emerging or reemerging and are transmitted by ticks, fleas, and lice vectors. In recent decades, there have been reports of rickettsias in ticks of the genus Amblyomma, Ixodes, and Haemaphysalis collected from wild birds. Accordingly, birds play a plausible role in the transport and spread of ticks infected by Rickettsia spp. In this study, we performed molecular detection of Rickettsia species in ticks collected from wild birds in the department of Caldas, Colombia. We detected and identified Rickettsia amblyommatis, 'Candidatus Rickettsia colombianensi' and a Rickettsia sp. closely related to 'Candidatus Rickettsia tarasevichiae' and Rickettsia canadensis. This study contributes to the knowledge on infection by Rickettsia in ticks collected from wild birds in Colombia. We also provide the first reports of infection by R. amblyommatis in the genus Ixodes collected from wild birds in South America and the presence of Rickettsia at elevations above 3000 m a.s.l.

Background. Cell-free DNA circulates in cancer patients and induces in vivo cell transformation and cancer progression in susceptible cells. Based on this, we hypothesized that depletion of circulating DNA with DNAse I and a protease mix could have antitumor effects. Study design. The study aimed to demonstrate that DNAse I and a protease mix can degrade in vitro DNA and proteins from the serum of healthy individuals and cancer patients, and in vivo in serum of Wistar rats,. Moreover, the antitumor effect of the systemically administered enzyme mix treatmentwas evaluated in nude mice subcutaneously grafted with the human colon cancer cell line SW480. Results. The serum DNA of cancer patients or healthy individuals was almost completely degraded in vitro by the enzymatic treatment, but no degradation was found with the enzymes given separately. The intravenous administration of the enzymes led to significant decreases in DNA and proteins from rat serum. No antitumor effect was observed in immunodeficient mice treated with the enzymes given separately. In contrast, the animals that received both enzymes exhibited a marked growth inhibition of tumors, 40% of them having pathological complete response. Conclusion. This study demonstrated that systemic treatment with DNAse I and a protease mix in rats decreases DNA and proteins from serum and that this treatment has antitumor effects. Our results support the hypothesis that circulating DNA could have a role in tumor progression, which can be offset by depleting it. Further studies are needed to prove this concept.

Ticks of the family Ixodidae are vectors of important pathogens in human and animal health. Birds are involved in long-distance transport and dispersion of hard ticks. Tick infestations on wild birds mostly involve species within the genera Amblyomma, Ixodes, and Haemaphysalis. In Colombia, tick research is scarce and there are no studies to date about the associations between wild birds and ticks. We aimed to contribute to the knowledge of the associations between wild birds and hard ticks based on the collection of 2314 wild birds belonging to 29 families in Caldas – Colombia between 2015 and 2019. In total, we collected 133 hard ticks that were found parasitizing 78 birds representing 45 species and 14 wild bird families. We report at least seven tick species on birds confirmed by morphological and molecular methods: Amblyomma longirostre, Amblyomma varium, Amblyomma dissimile, Amblyomma ovale, Amblyomma nodosum, Amblyomma calcaratum and Haemaphysalis leporispalustris. In addition, we recorded three Ixodes species, which yielded DNA sequences that did not have high identity (≤ 95 %) to any species in GenBank. Ticks were found infesting resident and migratory boreal birds. This is the first study addressing the associations between wild birds and hard ticks in Colombia. We describe new associations between birds and ticks in the Americas.

CD8+ T cell-mediated immune response plays a major role in the clearance of virus-infected cells, including human papillomavirus (HPV). The effective treatment of women with normal cytology but persistent high risk-HPV infection or with low-grade intraepithelial lesions could take advantage of novel strategies based on vaccination with viral immunological targets with a wide spectrum of cross-protection. The helicase E1, expressed early during viral replication in HPV infection, is among the most conserved papillomavirus proteins, which makes it a good vaccine candidate. In the present study, we examined E1-specific CD8+ T cell and NK immune responses in a mouse model with α-galactosylceramide (α-GalCer) as an adjuvant. We found that mice immunized with E1 combined with α-GalCer elicited an E1-specific CD8+ T and NK cell cytotoxic responses, which correlated with growth inhibition of grafted melanoma B16-F0 cells expressing E1, both in prophylactic and therapeutic protocols.

In tropical regions, the contributions and limitations of agroecosystems have been identified with respect to bird diversity conservation. It has been suggested that agroecosystems can have different conservation values, according to the structural complexity of the vegetation (e.g., higher number of vegetation strata, cultivated species diversity, among others). Therefore, agroecosystems, especially those with a smaller area (e.g., small-holdings), could be crucial for developing bird conservation strategies. In order to establish the contribution of different agroecosystem types to bird conservation in the department of Caldas (Colombia), we compared bird richness, abundance, and similarity associated to three types of habitats: (1) type I agroecosystems (mono-cropping with bare soil), (2) type II agroecosystems (mixed-cropping, grazing pastures with weeds and dispersed trees, and plantations with understory), and (3) secondary forests. Type II agroecosystems did not differ in bird richness and similarity compared to secondary forests, and species with high sensitivity to disturbance were registered (Zentrygon frenata, Phaetornis guy, Phaetornis syrmatophorus, Lepidocolaptes lacrymiger and Sphenopsis frontalis). Additionally, we registered a species of global conservation interest (Chloropipo flavicapilla) and four migratory species (Catharus ustulatus, Parkesia noveboracensis, Setophaga fusca and Setophaga striata) in this type of agroecosystem. Thus, type II agroecosystem habitats are not completely negative on avifauna, and they could serve an important role within conservation strategies in rural landscapes. En las regiones tropicales se han identificado los aportes y limitaciones de los agroecosistemas en el contexto de la conservación de la diversidad de las aves. Se ha sugerido que los agroecosistemas pueden presentar un valor diferente para la conservación, de acuerdo con la complejidad estructural de la vegetación (e.g., mayor número de estratos de la vegetación, diversidad de las especies cultivadas, entre otros). En este sentido, los agroecosistemas de una menor área (e.g., minifundios), pueden ser claves en el desarrollo de estrategias encaminadas a la conservación de la avifauna. Con el objetivo de establecer el aporte de diferentes tipos de agroecosistemas a la conservación de las aves en el departamento de Caldas (Colombia), se comparó la riqueza, abundancia y similitud de las aves asociadas a tres tipos de hábitats: (1) agroecosistemas tipo I (monocultivos con suelo limpio), (2) agroecosistemas tipo II (cultivos mixtos, potreros enmalezados con árboles dispersos y plantaciones con sotobosque) y (3) bosques secundarios. Los agroecosistemas tipo II no difirieron en la riqueza y en la similitud de las aves con respecto a los bosques secundarios, además se registraron especies con alta sensibilidad a la perturbación (Zentrygon frenata, Phaetornis guy, Phaetornis syrmatophorus, Lepidocolaptes lacrymiger y Sphenopsis frontalis). Adicionalmente en este tipo de agroecosistemas se registró una especie de interés para la conservación global (Chloropipo flavicapilla) y cuatro especies migratorias (Catharus ustulatus, Parkesia noveboracensis, Setophaga fusca y Setophaga striata). Los agroecosistemas tipo II no son hábitats completamente negativos para la avifauna y podrían desempeñar un rol importante dentro de las estrategias para la conservación en paisajes rurales.

Desde la promulgación de la Ley 100 del año 1993, existen en Colombia unas entidades de intermediación de la salud que en su momento se denominaron Empresas Promotoras de Salud (EPS), y que de acuerdo a la norma deberían de cumplir con las siguientes funciones: promoción de la afiliación de grupos no afiliados al sistema, captación de los aportes de los afiliados al sistema de seguridad social en salud, definir los mecanismos de acceso al sistema de salud por parte de los afiliados y sus familias, definir los protocolos que garanticen el libre acceso de los afiliados y sus familias a las instituciones prestadoras de salud, remitir al fondo de solidaridad y compensación la información relativa a la afiliación del trabajador y su familia, a las novedades laborales, a los recaudos por cotizaciones y a los desembolsos por el pago de prestación de servicios, y establecer los procedimientos para garantizar una atención oportuna, integral, eficiente y de calidad. Since the enactment of Law 100 in 1993, there have been some health intermediation entities in Colombia that at the time were called Health Promoting Companies (EPS), and that according to the norm should fulfill the following functions: promoting the affiliation of groups not affiliated with the system, capturing the contributions of affiliates to the social security health system, defining the mechanisms for access to the health system by affiliates and their families, defining the protocols that guarantee the Free access of the affiliates and their families to the health-providing institutions, to send to the solidarity and compensation fund the information regarding the affiliation of the worker and his family, to the labor news, to the collections for contributions and the disbursements for the payment provision of services, and establish procedures to guarantee timely, comprehensive, efficient and quality care.

La fasciolosis es una parasitosis que afecta rumiantes, equinos, porcinos, entre otras especies y es causante de grandes pérdidas económicas y puede parasitar humanos. En la presente investigación, se evaluó la frecuencia de la enfermedad por tres métodos de detección, en veintiséis predios, localizados en los municipios del departamento de Caldas, con mayor prevalencia, según reportes de la Central de beneficio de Manizales. Se utilizaron los métodos de coprología, PCR y ELISA, en heces y en suero de bovinos seleccionados aleatoriamente en predios, con el sistema de producción de leche bovina, localizados en los Andes colombianos, entre 1.980-3.721 msnm. Se obtuvieron 414 muestras de materia fecal, procesadas con la técnica Dennis modificada. El ADN fue extraído de las heces usando un kit comercial y fue amplificado con cebadores específicos, por PCR convencional. En las muestras de suero, se detectó la presencia de inmunoglobulina G (IgG) contra Fasciola hepatica, a través del uso de un Kit comercial, específico para este fin. La región estudiada, se dividió en cuatro zonas, para efectos del estudio. La prevalencia de la fasciolosis por coprología fue 12,3%; por Elisa, 19,1% y por PCR, 67,2%. La zona tres presentó la mayor proporción de animales positivos, por los tres métodos, con odds ratio elevados y alta significancia estadística; fueron hallados caracoles Limneidos en ambientes acuáticos, aptos para su desarrollo, que permiten perpetuar el inóculo. Se confirma la presencia de fasciolosis bovina en la región estudiada y se requieren hacer estudios adicionales, para validar la PCR, como método para la detección de fasciolosis. Fasciolosis is a parasitic disease that affects ruminants, horses, pigs and other species; it is the cause of significant economic losses and can also parasitize humans. In this research, the frequency of the disease using three detection methods in twenty-six farms, located in the municipalities most prevalent according to reports from the Slaughtering Central of Manizales-Caldas in the department of Caldas was evaluated. The coproscopic, PCR and ELISA methods in feces and serum of randomly selected cattle farms with a system of dairy production, located in the Colombian Andes, between 1980 3721m, were used. 414 stool samples were obtained and processed with the Dennis modified technique. DNA was extracted from stool using a commercial kit and was amplified with specific primers for standard PCR. In serum samples, the presence of immunoglobulin G (IgG) against Fasciola hepatica was detected through the use of a specific commercial kit for this purpose. The study area was divided into four zones for the study purposes. The prevalence of fascioliasis by the coproscopic test was 12.3%, by ELISA of 19.1%, and 67.2% by PCR. Zone three displayed the highest proportion of positive animals by the three methods, with high odds ratio and high statistical significance; lymnaeid snails were found in aquatic environments suitable for development, allowing perpetuate the inoculum. The presence of bovine fasciolosis in the studied region was confirmed; additional studies to validate PCR as a method by the fasciolosis detection are required. Incluye referencias bibliográficas

Natural Killer cells (NK) are key for the innate immune response against tumors and viral infections. Several viral proteins evade host immune response and target the NK cell receptor NKG2D and its ligands. Areas covered: This review aimed to describe the viruses and their proteins that interfere with the NKG2D receptor and their ligands, and how these interactions lead to immune evasion, host protection, and tissue damage from acute and chronic viral infections. Expert opinion: The study of viral proteins has already impacted the field of oncology. A prime example is the HBV vaccine and the development of antiviral drugs for HIV, Hepatitis C, and the family of Herpesviridae viruses. The NKG2D system seems to be a rational therapeutic target. Nevertheless, an effective cytotoxic response by NK cells is mediated by a network of activating and inhibitory receptors, the integration of which determines if the NK cell becomes cytotoxic or permissive. Immunotherapeutic agents that increase the antitumor lytic activity of NK cells through modulating activation and inhibitory signaling of NK cells are being developed. Nevertheless, more research is needed to dissect the integrative mechanism of NK cells function to fully exploit their antitumor and antiviral effector mechanisms.

A principios de este año se reportó por diferentes medios de comunicación una alerta naranja por la calidad del aire en Medellín. Este suceso hizo que el alcalde de la ciudad decretara unas medidas que evitaran que los niveles de contaminación ambiental siguieran empeorando y que los grupos de personas en mayor riesgo fueran afectados por esta contaminación. La calidad del aire es un problema que no se circunscribe a una región determinada, sino que es un problema mundial ya que el aire no tiene límites geográficos y dichos contaminantes pueden ser dispersados por la acción de los vientos, de modo que los desplazamientos de dichos contaminantes pueden afectar zonas menos contaminadas y de esta manera producir efectos dañinos no solo en las personas, sino también en las plantas y en los animales. A principios de este año se reportó por diferentes medios de comunicación una alerta naranja por la calidad del aire en Medellín. Este suceso hizo que el alcalde de la ciudad decretara unas medidas que evitaran que los niveles de contaminación ambiental siguieran empeorando y que los grupos de personas en mayor riesgo fueran afectados por esta contaminación. La calidad del aire es un problema que no se circunscribe a una región determinada, sino que es un problema mundial ya que el aire no tiene límites geográficos y dichos contaminantes pueden ser dispersados por la acción de los vientos, de modo que los desplazamientos de dichos contaminantes pueden afectar zonas menos contaminadas y de esta manera producir efectos dañinos no solo en las personas, sino también en las plantas y en los animales.

Purpose: The antihypertensive hydralazine has recently been repositioned as DNA demethylating for the epigenetic ther-apy of cancer. As the acetylator phenotype is the key determinant of its plasma levels, the dose of hydralazine needs to be adjusted for the acetylation status of patients. Methods: The pharmacokinetics of orally administered hydralazine was evaluated in 26 healthy vol-unteers (13 slow and 13 fast acetylators) af-ter a single dose of 182 mg administered as a controlled-release tablet. Plasma levels of hydralazine were analyzed in 85 cancer pa-tients treated with this formulation at a dose of 83 mg/d and 182 mg/d for slow and fast acetylators, respectively. Results: max The Cand tmax of hydralazine for fast acetylators were 208.4 ± 56.9 SD ng/ml and 2.8 ± 2.5 h, respectively. The corresponding results for slow acetylators were 470.4 ± 162.8 ng/ml, and 4.4 ± 3.1 h. Healthy volunteers who were fast acetylators had no clinically significant changes in blood pressure and heart rate or any other side-effect, however, slow acety-lators had transient episodes of headache, tachycardia and faintness. Among 85 cancer patients that received either 182 mg or 83 mg of hydralazine daily, according to their acetylator status, the mean concentrations of hydralazine in plasma were 239.1 ng/ml and 259.2 ng/ml for fast and slow acetylators, respectively. These differences were not sig-nificantly different, p = 0.3868. Conclusion: The administration of dose-adjusted con-trolled-release hydralazine according to the acetylation status of cancer patients yields similar levels of hydralazine.

The reversing of epigenetic aberrations using the inhibitors of DNA methylation and histone deacetylases may have therapeutic value in cervical cancer. This is a randomized phase III, placebo-controlled study of hydralazine and valproate (HV) added to cisplatin topotecan in advanced cervical cancer. Patients received hydralazine at 182 mg for rapid, or 83 mg for slow acetylators, and valproate at 30 mg/kg, beginning a week before chemotherapy and continued until disease progression. Response, toxicity, and PFS were evaluated, and 36 patients (17 CT + HV and 19 CT + PLA) were included. The median number of cycles was 6. There were four PRs to CT + HV and one in CT + PLA. Stable disease in five (29%) and six (32%) patients, respectively, whereas eight (47%) and 12 (63%) showed progression (P = 0.27). At a median follow-up time of 7 months (1-22), the median PFS is 6 months for CT + PLA and 10 months for CT + HV (P = 0.0384, two tailed). Although preliminary, this study represents the first randomized clinical trial to demonstrate a significant advantage in progression-free survival for epigenetic therapy over one of the current standard combination chemotherapy in cervical cancer. Molecular correlates with response and survival from this trial are pending to analyze.

Among many anticancer drugs collectively named "targeted or molecular therapies" epigenetic drugs are clearly promising. Differently from other agents targeting a single gene product, epigenetic drugs have chromatin as their target through inhibition of histone deacetylases (HDACs) and DNA methyltransferases (DNMTs) therefore, yet unspecific, they may act upon most or all tumor types, as deregulation of the methylation and deacetylation machinery are a common hallmark of neoplasia. In the last years, valproic acid (VPA) as emerged as a promising drug for cancer treatment. VPA has shown potent antitumor effects in a variety of in vitro and in vivo systems, and encouraging results in early clinical trials either alone or in combination with demethylating and/or cytotoxic agents. In addition, whole genome expression by microarray analysis from the primary tumors of patients treated with VPA show significant up-regulation of hundred of genes belonging to multiple pathways including ribosomal proteins, oxidative phosphorylation, MAPK signaling; focal adhesion, cell cycle, antigen processing and presentation, proteasome, apoptosis, PI3K, Wnt signaling, calcium signaling, TGF-beta signaling, and ubiquitin-mediated proteolysis among others. Despite in general, industry is not particularly interested in funding the clinical development of VPA, -at least in comparison to novel HDAC inhibitors-, existing preclinical and preliminary clinical data strongly suggest that VPA could be a drug that eventually will be used in combination therapies, either with classical cytotoxics, other molecular-targeted drugs or radiation in a number of solid tumors.

Background: Epigenetic aberrations lead to chemotherapy resistance; hence, their reversal by inhibitors of DNA methylation and histone deacetylases may overcome it. Patients and methods: Phase II, single-arm study of hydralazine and magnesium valproate added to the same schedule of chemotherapy on which patients were progressing. Schedules comprised cisplatin, carboplatin, paclitaxel, vinorelbine, gemcitabine, pemetrexed, topotecan, doxorubicin, cyclophosphamide, and anastrozole. Patients received hydralazine at 182 mg for rapid, or 83 mg for slow, acetylators, and magnesium valproate at 40 mg/kg, beginning a week before chemotherapy. Response, toxicity, DNA methylation, histone deacetylase activity, plasma valproic acid, and hydralazine levels were evaluated. Results: Seventeen patients were evaluable for toxicity and 15 for response. Primary sites included cervix (3), breast (3), lung (1), testis (1), and ovarian (7) carcinomas. A clinical benefit was observed in 12 (80%) patients: four PR, and eight SD. The most significant toxicity was hematologic. Reduction in global DNA methylation, histone deacetylase activity, and promoter demethylation were observed. Conclusions: The clinical benefit noted with the epigenetic agents hydralazine and valproate in this selected patient population progressing to chemotherapy’ and re-challenged with the same chemotherapy schedule after initiating hydralazine and valproate’ lends support to the epigenetic-driven tumor-cell chemoresistance hypothesis (ClinicalTrials.gov Identifier: NCT00404508).

Persistent infections of the uterine cervix with ‘high-risk’ human papillomavirus (HPV) are now recognized as necessary for the development of cervical cancer. Among them, HPV types 16 and 18 exhibit numerous variants associated with different risks for cervical cancer development. In this study, the questions of whether different HPV type 18 variants exhibit changes in early gene transcription and the molecular mechanisms underlying these differences were investigated. It was shown that, indeed, type 18 variants exhibited singular differences in E6 transcripts in vivo. Higher levels of the E6*I transcript were detected regularly in clones harbouring the African variant, as opposed to low levels of this transcript detected in clones containing the reference clone (Asian–Amerindian), where significantly higher levels of full-length E6 transcript were usually observed. As a direct consequence, higher levels of p53 protein were found in the presence of African E6, as opposed to the low levels of p53 observed with the Asian–Amerindian E6. These variations in consequence affected the levels of cellular proteins regulated by p53, such as Bax. Similar changes in the relative levels of E6 transcripts were observed when tumours containing type 18 E6 variants were analysed. The different ability of cells containing variant E6 genes to form tumours in nude mice was suggested by the fact that tumour volumes were considerably higher when cells expressed the Asian–Amerindian E6. Mutagenesis analysis of the reference clone showed that a C491A change reverts the phenotype. These results suggest that different splicing patterns of E6 within HPV type 18 variants may possibly have biological implications in viral tumorigenesis.

Ether à go-go (EAG) potassium channels display oncogenic properties. In normal tissues, EAG mRNA is almost exclusively expressed in brain, but it is expressed in several somatic cancer cell lines, including HeLa, from cervix. Antisense experiments against eag reduce cell proliferation in some cancer cell lines, and inhibition of EAG-mediated currents has been suggested to decrease cell proliferation in a melanoma cell line. Because of the potential clinical relevance of EAG, we investigated EAG mRNA expression in the following fresh samples from human uterine cervix: 5 primary cultures obtained from cancerous biopsies, 1 cancerous fresh tissue, and 12 biopsies of control normal tissue. All of the control cervical samples came from patients with negative pap smears. Reverse transcription-PCR and Southern-blot experiments revealed eag expression in 100% of the cancerous samples and in 33% of the normal biopsies. Immunochemistry experiments showed the presence of EAG channel protein in cells from the primary cultures and in cervical cancer biopsies sections from the same patients. In addition, we looked for EAG-mediated currents in the cultures from cervical cancer cells. Here we show for the first time EAG channel activity in human tumors. Patch-clamp recordings showed typical EAG-mediated currents modulated by magnesium and displaying a pronounced Cole-Moore shift. Because EAG expression and channel activity have been suggested to be important in cell proliferation, our findings strongly support the idea of considering EAG as a tumor marker as well as a potential membrane therapeutic target for cervical cancer.

Elevated expression of matrix metalloproteinases (MMPs) plays a critical role in extracellular matrix (EM) degradation in tumor development and prognosis of different human carcinomas. In cervical carcinoma (Ce Ca), the role of these proteinases in the biological development of this neoplasm is controversial. In the present study, we compared the secretion of MMP-2, MMP-3 and MMP-9 among 29 benign and premalignant cervical lesions (cervicitis and cervical intraepithelial neoplasias) and 46 tumoral explants of Ce Ca. The explants were cultured for 48 h. The gelatinases secreted into conditioned medium were revealed by zymography and quantified by densitometry. The results showed high levels of MMP-3 and MMP-9 in tumoral explants. In contrast, only the pro-MMP-2 was higher in benign cervical lesions, although both active and inactive MMP-2 species are associated with advanced clinical stages in tumoral samples, and only the secretion of MMP-3 was associated with unresponsiveness to radiotherapy. We can conclude that the expression of MMPs is related to the invasive process in Ce Ca and suggest that they may play a role in degradation of the EM during local invasion. In addition, MMP-3 secretion could be a marker of poor prognosis in Ce Ca.

It is known that cell-free DNA circulates in plasma/serum of patients with cancer and that part of this DNA circulates as nucleosomes that can be quantified by ELISA. We analyzed the effect of tumor and chemotherapy upon the levels of nucleosomes in vitro, in vivo and in cervical cancer patients. The levels of nucleosomes pre- and post-treatment were correlated with response in 11 patients receiving chemotherapy. Nucleosomes were determined in nude mice treated with or without cisplatin and carrying tumors generated with HeLa cells, and in the cell lysate and supernatant of HeLa cells exposed to cisplatin in culture. In addition, nucleosomes were determined at different time points in patients and in rats receiving chemotherapy. Nucleosomes were higher in patients that controls (1,760 vs. 601, p = 0.0001). After 24 hr of treatment with oxaliplatin and gemcitabine, the levels decreased in 6 patients of whom 5 had response. Nucleosome levels differed between mice xenografted and not xenografted (765 vs. 378, p = 0.001) and between xenografted treated with or without cisplatin (650 vs. 765, p = 0.010), but not in tumor-free animals treated and untreated with cisplatin (378 vs. 379, p = 0.99). In vitro, nucleosomes reached at peak 8 hr in cell lysates to decrease thereafter, whereas in supernatant, levels continued to increase up to 24 hr. Serial determination of nucleosomes in patients showed a rise within 6-12 hr and then a reduction to below the basal at 24 hr. In rats, nucleosomes had no major changes in those receiving oxaliplatin or the triple combination of cisplatin, gemcitabine and paclitaxel as compared to untreated controls. An overdose of this triple combination produced a transient elevation of almost 1,000 AU over the basal. Our results demonstrate that most of circulating nucleosomes originate from the tumor and that chemotherapy produces an early rise most likely due to tumor apoptosis and that nucleosomes are rapidly cleared from circulation. On the contrary, chemotherapy within the therapeutic range of doses has no effect on nucleosome levels in healthy mice and rats. This data suggests that the determination of circulating nucleosomes pre- and post-treatment could be a useful test to predict response to chemotherapy in cancer patients.

The aim of the present study was to demonstrate that ribavirin, a known inhibitor of eIF4E and inosine 5'-phosphate dehydrogenase (IMPDH), also inhibits histone methyltransferase zeste homolog 2 (EZH2). A computational searching revealed that ribavirin has a high structural similarity to 3-deazaneplanocin A (DZNep). The growth inhibitory effects of ribavirin as well as its effects upon epigenetic enzymes were evaluated in various cancer cell lines. siRNA assays were used to downregulate eIF4E, EZH2 and IMPDH to determine the contribution of these targets to the growth inhibitory effects of ribavirin. Ribavirin decreased EZH2 expression, inhibited histone methyltransferase activity and decreased H3K27 trimethylation. Ribavirin induced variable growth inhibition in a number of cell lines and downregulation of the targets, EZH2, eIF4E and IMPDH1 and 2 by siRNA led to comparable growth inhibition while no significant further reduction in viability was observed when siRNA transfected cells were treated with ribavirin. The results showed that ribavirin inhibits these cancer targets and should thus be studied for cancer therapy.

Most cancer deaths are due to metastases. Metastasis is an extraordinarily complex process by which cancer cells complete a sequential series of steps before they transform into a clinically detectable lesion. These steps typically include separation from the primary tumor, invasion through surrounding tissues and basement membranes, entry and survival in the circulation, lymphatic or peritoneal space, and arrest in a distant target organ and the formation of secondary tumors in distant organs.While proposed or accepted models and mechanisms of metastatic progression, have been demonstrated in experimental systems, none of them sufficiently explain all of the complexities associated with this process. These models can broadly be classified into two types, those occurring by vertical gene transfer (Darwinian) and those involving horizontal or lateral DNA transfer. Here, we describe an experimental system to study the metastatic process involving the horizontal transfer of circulating DNA.

Most cancer deaths are due to metastases. Metastasis is an extraordinarily complex process by which cancer cells complete a sequential series of steps before they transform into a clinically detectable lesion. These steps typically include separation from the primary tumor, invasion through surrounding tissues and basement membranes, entry and survival in the circulation, lymphatic or peritoneal space, and arrest in a distant target organ and the formation of secondary tumors in distant organs.While proposed or accepted models and mechanisms of metastatic progression, have been demonstrated in experimental systems, none of them sufficiently explain all of the complexities associated with this process. These models can broadly be classified into two types, those occurring by vertical gene transfer (Darwinian) and those involving horizontal or lateral DNA transfer. Here, we describe an experimental system to study the metastatic process involving the horizontal transfer of circulating DNA.

Adenocarcinomas arising in the surface epithelium of nasal and paranasal cavities (NPCA) are uncommon. There are 2 types: a) low-grade adenocarcinomas, and b) intestinal-type adenocarcinomas.

BACKGROUND: Down regulation of genes coding for nucleoside transporters and drug metabolism responsible for uptake and metabolic activation of the nucleoside gemcitabine is related with acquired tumor resistance against this agent. Hydralazine has been shown to reverse doxorubicin resistance in a model of breast cancer. Here we wanted to investigate whether epigenetic mechanisms are responsible for acquiring resistance to gemcitabine and if hydralazine could restore gemcitabine sensitivity in cervical cancer cells. METHODOLOGY/PRINCIPAL FINDINGS: The cervical cancer cell line CaLo cell line was cultured in the presence of increasing concentrations of gemcitabine. Down-regulation of hENT1 & dCK genes was observed in the resistant cells (CaLoGR) which was not associated with promoter methylation. Treatment with hydralazine reversed gemcitabine resistance and led to hENT1 and dCK gene reactivation in a DNA promoter methylation-independent manner. No changes in HDAC total activity nor in H3 and H4 acetylation at these promoters were observed. ChIP analysis showed H3K9m2 at hENT1 and dCK gene promoters which correlated with hyper-expression of G9A histone methyltransferase at RNA and protein level in the resistant cells. Hydralazine inhibited G9A methyltransferase activity in vitro and depletion of the G9A gene by iRNA restored gemcitabine sensitivity. CONCLUSIONS/SIGNIFICANCE: Our results demonstrate that acquired gemcitabine resistance is associated with DNA promoter methylation-independent hENT1 and dCK gene down-regulation and hyper-expression of G9A methyltransferase. Hydralazine reverts gemcitabine resistance in cervical cancer cells via inhibition of G9A histone methyltransferase.

It is known that cancer progresses by vertical gene transfer, but this paradigm ignores that DNA circulates in higher organisms and that it is biologically active upon its uptake by recipient cells. Here we confirm previous observations on the ability of cell-free DNA to induce in vitro cell transformation and tumorigenesis by treating NIH3T3 recipient murine cells with serum of colon cancer patients and supernatant of SW480 human cancer cells. Cell transformation and tumorigenesis of recipient cells did not occur if serum and supernatants were depleted of DNA. It is also demonstrated that horizontal cancer progression mediated by circulating DNA occurs via its uptake by recipient cells in an in vivo model where immunocompetent rats subjected to colon carcinogenesis with 1,2-dimethylhydrazine had increased rate of colonic tumors when injected in the dorsum with human SW480 colon carcinoma cells as a source of circulating oncogenic DNA, which could be offset by treating these animals with DNAse I and proteases. Though the contribution of biologically active molecules other than DNA for this phenomenon to occur cannot be ruled out, our results support the fact that cancer cells emit into the circulation biologically active DNA to foster tumor progression. Further exploration of the horizontal tumor progression phenomenon mediated by circulating DNA is clearly needed to determine whether its manipulation could have a role in cancer therapy.

Natural killer cells play a role in the immune antitumor response by recognizing and eliminating tumor cells through the engagement of NKG2D receptors with their ligands on target cells. This work aimed to investigate whether epigenetic drugs are able to increase MICA and MICB expression as well as NK cell cytotoxicity. Prostate, colon, breast and cervical cancer cell lines were analyzed for the expression of MICA and MICB at the mRNA and protein levels by RT-PCR, Western blot, flow cytometry and ELISA. The activating mark H3K4m2 at the MICA and MICB promoters was investigated by ChIP assays. Cytotoxicity of NK cells against the target epithelial cancer cells was investigated with the CD107 cytotoxicity assay. The results show that hydralazine and valproic acid not only increase the expression of MICA and MICB ligands of target cells, but also reduce their shedding to the supernatant. This upregulation occurs at the transcriptional level as revealed by increase of the H3K4 activating mark at the promoter of MICA and MICB genes. These effects are paralleled by increased cytotoxicity of NK cells, which was attenuated at different degrees by using blocking antibodies against the NKG2D receptor and ligands. In conclusion, our results demonstrate the ability of hydralazine and valproate to increase the NK activity against epithelial cancer cell lines and suggest that these drugs could reduce the levels of soluble MICA and MICB helping in avoiding tumor-induced suppression of NK cytotoxicity against the tumor.

Aberrant DNA methylation and histone deacetylation participate in cancer development and progression; hence, their reversal by inhibitors of DNA methylation and histone deacetylases is a promising cancer therapy. Experimental data demonstrate that these inhibitors in combination do not only show synergy in antitumor effects but also in whole genome global expression. Ten pairs of pre- and post-treatment cervical tumor samples were analyzed by microarray analysis. Treatment for seven days with hydralazine and valproate (HV) in patients up-regulated 964 genes. The two pathways possessing the highest number of up-regulated genes comprised the ribosome protein and the oxidative phosphorylation pathways, followed by MAPK signaling, tight junction, adherens junction, actin cytoskeleton, cell cycle, focal adhesion, apoptosis, proteasome, Wnt signaling, and antigen processing and presentation pathways. Up-regulated genes by HV, clustered with down-regulated genes in untreated primary cervical carcinomas and were more alike as compared with up-regulated genes from untreated patients in terms of gene ontology. Increased acetylated p53 was also observed. Epigenetic therapy with HV leads to gene reactivation in primary tumors of cervical cancer patients as well as protein acetylation. A number of these reactivated genes have a definitive role as a tumor suppressors. The global expression pattern induced by HV suggests this therapy has an impact on pathways related to energy production which may promote apoptosis.

Decitabine and azacitidine, two DNA methyltransferase (DNMT) inhibitors, are the current standard of treatment for myelodysplastic syndrome (MDS). Histone deacetylase (HDAC) inhibitors are also being tested against MDS. Both drug classes synergize in their gene reactivating and anticancer activities. The combination of hydralazine and valproate (Transkrip®), a DNMT and HDAC inhibitor, respectively), has been developed as epigenetic therapy under the drug repositioning concept. To evaluate the clinical efficacy and safety of hydralazine and valproate against MDS, an open phase-II study for previously treated patients with MDS was conducted. The hydralazine dose was given according with the acetylator phenotype, and valproate was dosed at 30 mg/kg/day. Response was graded with International Working Group criteria. Toxicity was evaluated by the Common Toxemia Criteria-National Cancer Institute version 3 scale. From November 2007 to January 2010, 12 patients were included. Median age ± SD was 53 ± 19.78 years (range, 23–79 years); median time from diagnosis to inclusion in the study was 7.9 months (range 2.6–36.1 months). Median of previous treatment was 2 (range, 1–6). Refractory cytopenia with multilineage dysplasia was diagnosed in ten cases, and refractory anemia with excess of blasts in two. Overall response was documented in six (50%) of 12 cases, including one CR, one PR, and four hematological improvements of the erythroid series. Two patients (16.6%) progressed to acute myeloid leukemia. Hemoglobin increased from 7.4 to 10.3 g/dL (in 13 weeks), neutrophils, from 1.1 to 2.0 (in 3 weeks), and platelets, from 66 × 109 to 72 × 109/L (in 2 weeks). Transfusional requirements decreased from 2.3 to 0 U bi-monthly for red blood cells and from 0.5 to 0 U bi-monthly for platelets in responding patients. Main toxicities were mild, including somnolence and nausea. Preliminary results of this phase-II study suggest that the combination of hydralazine and valproate is a promising non-toxic and effective therapy for MDS.

Background Germ-line mutations of the TP53 gene are known to cause Li-Fraumeni syndrome, an autosomal, dominantly inherited, high-penetrance cancer-predisposition syndrome characterized by the occurrence of a variety of cancers, mainly soft tissue sarcomas, adrenocortical carcinoma, leukemia, breast cancer, and brain tumors. Methods Mutation analysis was based on Denaturing high performance liquid chromatography (DHPLC) screening of exons 2-11 of the TP53 gene, sequencing, and cloning of DNA obtained from peripheral blood lymphocytes. Results We report herein on Li Fraumeni syndrome in a family whose members are carriers of a novel TP53 gene mutation at exon 4. The mutation comprises an insertion/duplication of seven nucleotides affecting codon 110 and generating a new nucleotide sequence and a premature stop codon at position 150. With this mutation, the p53 protein that should be translated lacks the majority of the DNA binding domain. Conclusion To our knowledge, this specific alteration has not been reported previously, but we believe it is the cause of the Li-Fraumeni syndrome in this family.

Gemcitabine (2′,2′-difluoro 2′deoxycytidine, dFdC) is an analog of cytosine with distinctive pharmacological properties and a wide antitumor-activity spectrum. The pharmacological characteristics of gemcitabine are unique because two main classes of genes are essential for its antitumor effects: membrane transporter protein-coding genes, whose products are responsible for drug intracellular uptake, as well as enzyme-coding genes, which catalyze its activation and inactivation. The study of the pharmacogenetics and pharmacoepigenetics of these two gene classes is greatly required to optimize the drug’s therapeutic use in cancer. This review aims to provide an update of genetic and epigenetic bases that may account for interindividual variation in therapeutic outcome exhibited by gemcitabine.

Background The methylation status at the human papilloma virus (HPV) genome found in pre-invasive and invasive cervical lesions suggests that neoplastic transformation can be suppressed by gene hypermethylation, whereas hypomethylation accompanies or causes cancer progression; hence, epigenetic therapy aimed at reactivating cellular suppressor-gene expression has the potential to act as a tumor promoter by enhancing HPV oncoprotein expression in HPV-related malignancies. The objective of this study was to determine the influence of hydralazine and valproate on HPV oncogene expression in cervical cancer cell lines and the primary tumors of patients undergoing treatment with hydralazine and valproate. Results Overall, hydralazine and valproate either alone or combined exerted a growth inhibitory effect on cervical cancer cell lines. A cell line-specific up-regulating effect was observed on E6/E7 gene expression, which in general correlated with DNA hypomethylation and histone acetylation at the long control region (LCR). Nonetheless, E6/E7 expression was unchanged or decreased in the majority of patients with cervical cancer treated with hydralazine, valproate, or both. In some cervical cancer cell lines, these drugs led to increased transcription of p53, and increased its stabilization due to acetylation at lysines 273 and 282, which allowed a higher bax-protein transactivating effect. Conclusion The results of this study demonstrate that hydralazine and valproate can be safely administered to HPV-related malignancies such as cervical cancer because they do not increase viral oncoprotein expression. Most importantly, the antitumor effect of hydralazine and valproate in cervical cancer may at least partially depend on an up-regulating effect on p53 gene and on the valproate-induced hyperacetylation of p53 protein, protecting it from degradation by E6.