Plasma membrane lipid bilayer is druggable: Selective delivery of gemcitabine-squalene nano-medicine to cancer cells.

Up to now the lipid bilayers were rarely considered as targets in cancer therapy despite pronounced differences in lipid composition between plasma membranes of benign and malignant cells. In this study we demonstrate that the lipid bilayer of the plasma membrane is druggable and suitable for facilitating selective delivery of amphiphilic gemcitabine-squalene nanomedicines to cancer cells. Data from radioactive assays, fluorescent membrane probes and molecular dynamics simulations provide evidence of selective accumulation of gemcitabine-squalene in the plasma membranes with disrupted lipid asymmetry and its subsequent preferential uptake by malignant cells. This causes pronounced cytotoxicity on cancer cells in comparison to their benign counterparts originating from the same tissue.

Wound Healing versus Metastasis: Role of Oxidative Stress.

Many signaling pathways, molecular and cellular actors which are critical for wound healing have been implicated in cancer metastasis. These two conditions are a complex succession of cellular biological events and accurate regulation of these events is essential. Apart from inflammation, macrophages-released ROS arise as major regulators of these processes. But, whatever the pathology concerned, oxidative stress is a complicated phenomenon to control and requires a finely tuned balance over the different stages and responding cells. This review provides an overview of the pivotal role of oxidative stress in both wound healing and metastasis, encompassing the contribution of macrophages. Indeed, macrophages are major ROS producers but also appear as their targets since ROS interfere with their differentiation and function. Elucidating ROS functions in wound healing and metastatic spread may allow the development of innovative therapeutic strategies involving redox modulators.

Estudio fitoquimico, actividad antioxidante y toxicidad sobre Artemia salina de los extractos de las hojas de Justicia secunda Vahl. (Acanthaceae), recolectada en Merida-Venezuela

Objetivos: realizar el estudio fitoquímico cualitativo para determinar la actividad antioxidante y toxicidad sobre Artemia salina de los extractos de las hojas de Justicia secunda Vahl, recolectada en Mérida, Venezuela. Metodología: el tamizaje fitoquímico de los extractos de las hojas de Justicia secunda Vahl. (Acanthaceae) se realizó a través de pruebas químicas específicas y permitió determinar la presencia de este-roles en el extracto hexanoico; esteroles y compuestos fenólicos en el diclorometanoico; alcaloides, esteroles y compuestos fenólicos en el extracto etanólico. La actividad antioxidante de los extractos de J. secunda fue evaluada usando el método de la capacidad secuestrante de radicales libres de 2,2-difenil-1-picrilhidrazilo (DPPH») mediante espectrofotometría UV-visible a una longitud de onda de 517 nm, con ácido ascórbico como control positivo (176 µg/rnL). Resultados: los extractos diclorometanoico y etanólico mostraron actividad antioxidante, con un porcentaje de inhibición superior a 50 a 4 mg/mL y a una concentración de 0,75 mg/mL un porcentaje de inhibición de 27,7 y 53,0; IC50 de 1,85 mg/mL y 0,69 mg/mL, respectivamente. En cuanto a la toxicidad sobre A. salina, del extracto etanólico mostró una DL50 entre 919,32 y 3781,9 ppm y es relativamente inocuo, según la clasificación CYTED. Conclusión: en general, los resultados obtenidos sugieren que los extractos de las hojas de J. secunda Vahl, podrían ser una alternativa en la formulación de fármacos, debido a su potencial como antioxidante y su baja toxicidad.

SUMMARY Aims: To carry out the qualitative phytochemical study to determine the antioxidant activity and toxicity on Artemia salina of the extracts of the leaves of Justicia secunda Vahl, collected in Mérida, Venezuela. Methodology: Phytochemical screening of extracts from the aerial parts of Justicia secunda Vahl. (Acanthaceae) was carried out through specific chemical tests that allowed the presence of sterols in the hexanoic extract to be determined; sterols and phenolic compounds in the dichloromethane and alkaloids, sterols and phenolic compounds in the ethanolic extract. The antioxidant activity of extracts from the aerial parts of J. secunda was evaluated by using the 1,1-diphenyl-2-picrylhydrazyl (DPPH•) radical scavenging method, using spectrophotometry UV-Visible at a wavelength of 517 nm, with ascorbic acid as control (176 µg/mL). Results: The dichloromethane and ethanolic extracts showed anti-oxidant activity, with an inhibition percentage higher than 50 to 4 mg/mL and at a concentration of 0.75 mg / mL, percentage of 27.7 and 53.0; IC50 of 1.85 mg/mL and 0.69 mg/mL, respectively. Regarding toxicity on A. salina, the ethanolic extract showed an LD50 between 919.32 and 3781.9 ppm, it was relatively harmless, according to the CYTED classification. Conclusion: Overall, the results obtained suggest that the extracts of the aerial parts of J. secunda Vahl, could be an alternative for future drug formulation since it has potent antioxidant activity and low toxicity.

Objetivo: realizar o estudo fitoquímico qualitativo para determinar a atividade antioxidante e toxicidade sobre Artemia salina dos extratos das folhas de Justicia secunda Vahl, coletadas em Mérida, Venezuela. Metodologia: triagem fitoquímica de extratos das partes aéreas de Justicia secunda Vahl. (Acanthaceae) foi realizado por meio de testes químicos específicos e permitiu determinar a presença de esteróis no extrato hexanoico; esteróis e compostos fenólicos no diclorometano e alcalóides, esteróis e compostos fenólicos no extrato etanólico. A atividade antioxidante de extratos das partes aéreas de J. secunda foi avaliada pelo método de sequestro do radical 1,1-difenil-2-picrilhidrazil (DPPH»), por espectrofotometria UV-Visível no comprimento de onda de 517 nm, com ácido ascórbico como controle (176 µg/mL). Resultados: os extratos diclorometano e etanólico apresentaram atividade antioxidante, com percentual de inibição superior a 50 a 4 mg/mL e na concentração de 0,75 mg/ mL, percentagem de 27,7 e 53,0; IC50 de 1,85 mg/mL e 0,69 mg/mL, respectivamente. Em relação à toxicidade sobre A. salina, o extrato etanólico apresentou DL50 entre 919,32 e 3781,9 ppm, sendo relativamente inofensivo, segundo a classificação CYTED. Conclusão: de maneira geral, os resultados obtidos sugerem que os extratos das partes aéreas de J. secunda Vahl, podem ser uma alternativa para futura formulação de fármacos por apresentarem potente atividade antioxidante e baixa toxicidade.

Leptin-Induced HLA-G Inhibits Myometrial Contraction and Differentiation.

Maternal obesity is associated with a wide spectrum of labour disorders, including preterm birth. Leptin, a pro-inflammatory adipokine and a key factor of obesity, is suspected to play a major role in these disorders. OB-R, its receptor, is expressed on macrophages and myocytes, two cell types critical for labour onset. Macrophages secrete reactive oxygen species/pro-inflammatory cytokines, responsible for myometrial differentiation while myocytes control uterine contractions. In this study, we assessed the effect of leptin on myometrial contraction and differentiation using our validated co-culture model of human primary macrophages and myocytes. We demonstrated that leptin had a different effect on myocytes and macrophages depending on the dose. A low leptin concentration induced a tocolytic effect by preventing myocytes' contraction, differentiation, and macrophage-induced ROS production. Additionally, leptin led to an increase in HLA-G expression, suggesting that the tocolytic effect of leptin may be driven by HLA-G, a tolerogenic molecule. Finally, we observed that recombinant HLA-G also prevented LPS-induced ROS production by macrophages. Altogether, these data provide a putative molecular mechanism by which leptin may induce immune tolerance and therefore interfere with labour-associated mechanisms. Therefore, HLA-G represents a potential innovative therapeutic target in the pharmacological management of preterm labour.

Caracterización sociodemográfica, clínica y terapéutica de pacientes con falla cardiaca con fracción de eyección intermedia: cohorte MED-ICAi / Sociodemographic, clinical and therapeutic characteristics of patients with heart failure with mid-range ejection fraction: MED-ICAi cohort

Resumen Antecedentes: Clásicamente, la falla cardiaca se ha clasificado en dos grupos, según tengan fracción de eyección preservada o reducida; no obstante, en fecha reciente se ha añadido el grupo de fracción intermedia, y aún existe gran desconocimiento sobre sus características fisiopatológicas y clínicas. Objetivo: Caracterizar el grupo de pacientes con fracción intermedia en cuanto a sus variables sociodemográficas, clínicas y de tratamiento. Método: Se realizó un estudio descriptivo, retrospectivo, en el que se analizaron historias clínicas de pacientes con falla cardiaca hospitalizados por agudización en el periodo comprendido entre enero de 2015 y diciembre de 2017. Resultados: Se revisaron 1536 historias clínicas, de las cuales 864 cumplían los criterios de inclusión. El grupo con fracción intermedia correspondió a 83 (9.6%) pacientes, en quienes se encontró predominio del sexo femenino (53%) y una edad mediana de 77 años. La coronariopatía fue la etiología más frecuente (26.5%), mientras que la falta de adherencia a los medicamentos fue la causa principal de descompensación (14.5%). Los medicamentos más usados fueron los betabloqueadores y la furosemida, tanto al ingreso como al egreso. El grupo con mayor mortalidad fue el de fracción de eyección reducida (4.1%). La estancia hospitalaria, el ingreso y la estancia en la unidad de cuidados intensivos fueron similares en todos los grupos, independientemente de la fracción de eyección. Conclusiones: Los hallazgos son similares a los descritos en otras poblaciones internacionales y en algunas nacionales, y avalan la hipótesis de un fenotipo intermedio con un comportamiento etiológico semejante al de la fracción de eyección reducida.

Abstract Background: Classically, heart failure has been classified in two groups, depending on a preserved or reduced ejection fraction, but a mid-range ejection fraction group has been introduced recently, and there is still great ignorance about its physiopathological and clinical characteristics. Objective: To characterize this group of patients as for their sociodemographic, clinical and treatment variables. Method: We carried out a descriptive, retrospective study, by analyzing medical records from patients hospitalized with acute heart failure between January 2015 and December 2017. Results: We reviewed 1536 medical records of which 864 met the inclusion criteria. The mid-range ejection fraction group corresponded to 83 (9.6%) of patients, of which the majority were women (53%), with a median age of 77 years, coronary heart disease as the most frequent etiology (26.5%) and lack of adherence to medications as the main cause of decompensation (14.5%). The most frequently used drugs were betablockers and furosemide, both upon admission and discharge. Mortality was higher between patients with reduced ejection fraction (4.1%). Hospital stay, admission to and length of stay in an ICU, were similar between all groups regardless of ejection fraction. Conclusions: Our findings are similar to those described in previous international and national cohorts, and support the hypothesis of an intermediate phenotype with an etiology similar to that seen with a reduced ejection fraction.

Lactobacillus stress protein GroEL prevents colonic inflammation.

BACKGROUND: We previously showed that supernatants of Lactobacillus biofilms induced an anti-inflammatory response by affecting the secretion of macrophage-derived cytokines, which was abrogated upon immunodepletion of the stress protein GroEL. METHODS: We purified GroEL from L. reuteri and analysed its anti-inflammatory properties in vitro in human macrophages isolated from buffy coats, ex vivo in explants from human biopsies and in vivo in a mouse model of DSS induced intestinal inflammation. As a control, we used GroEL purified (LPS-free) from E. coli. RESULTS: We found that L. reuteri GroEL (but not E. coli GroEL) inhibited pro-inflammatory M1-like macrophages markers, and favored M2-like markers. Consequently, L. reuteri GroEL inhibited pro-inflammatory cytokines (TNFα, IL-1ß, IFNγ) while favouring an anti-inflammatory secretome. In colon tissues from human biopsies, L. reuteri GroEL was also able to decrease markers of inflammation and apoptosis (caspase 3) induced by LPS. In mice, we found that rectal administration of L. reuteri GroEL (but not E. coli GroEL) inhibited all signs of haemorrhagic colitis induced by DSS including intestinal mucosa degradation, rectal bleeding and weight loss. It also decreased intestinal production of inflammatory cytokines (such as IFNγ) while increasing anti-inflammatory IL-10 and IL-13. These effects were suppressed when animals were immunodepleted in macrophages. From a mechanistic point of view, the effect of L. reuteri GroEL seemed to involve TLR4, since it was lost in TRL4-/- mice, and the activation of a non-canonical TLR4 pathway. CONCLUSIONS: L. reuteri GroEL, by affecting macrophage inflammatory features, deserves to be explored as an alternative to probiotics.

Lipoproteins LDL versus HDL as nanocarriers to target either cancer cells or macrophages.

In this work, we have explored natural unmodified low- and high-density lipoproteins (LDL and HDL, respectively) as selective delivery vectors in colorectal cancer therapy. We show in vitro in cultured cells and in vivo (NanoSPECT/CT) in the CT-26 mice colorectal cancer model that LDLs are mainly taken up by cancer cells, while HDLs are preferentially taken up by macrophages. We loaded LDLs with cisplatin and HDLs with the heat shock protein-70 inhibitor AC1LINNC, turning them into a pair of "Trojan horses" delivering drugs selectively to their target cells as demonstrated in vitro in human colorectal cancer cells and macrophages, and in vivo. Coupling of the drugs to lipoproteins and stability was assessed by mass spectometry and raman spectrometry analysis. Cisplatin vectorized in LDLs led to better tumor growth suppression with strongly reduced adverse effects such as renal or liver toxicity. AC1LINNC vectorized into HDLs induced a strong oxidative burst in macrophages and innate anticancer immune response. Cumulative antitumor effect was observed for both drug-loaded lipoproteins. Altogether, our data show that lipoproteins from patient blood can be used as natural nanocarriers allowing cell-specific targeting, paving the way toward more efficient, safer, and personalized use of chemotherapeutic and immunotherapeutic drugs in cancer.

Gonadal maturation in Pseudoplatystoma metaense x Leiarius marmoratus hybrids, (Siluriformes: Pimelodidae) / Maduración gonadal en híbridos de Pseudoplatystoma metaense x Leiarius marmoratus (Siluriformes: Pimelodidae)

SUMMARY: Establishing guidelines for responsible management in fish production systems requires knowledge of the basic biology of the fish to be raised. The objective of this work was to determine the reproductive capacity of hybrids produced from the crossing of females of Pseudoplatystoma metaense with males of Leiarius marmoratus. Males presented a digitiform, unrestricted spermatogonial testicle containing caudal digits producing glycoproteins that do not form a seminal vesicle. It was possible to find free sperm in the lumen of the tubules and in the ducts. The ovary of the females was found to be saccular and synchronous with at least three groups of oocytes. In the first year of life, only oogonia up to previtellogenic oocytes (cortical alveolus 284.9 ± 35.7 mm in diameter) were found. After the second year vitellogenic oocytes 730 ± 3.78 mm in diameter were observed. The events of gonadal development of the hybrids indicate that they are gonochoric, synchronic animals. The maturation peaks in the high-water season, overlapping with the parental species. Therefore, the escape of hybrids from fish cultures to the rivers may increase the risk of crosses, gene introgression, or diminution of the reproductive capacity of the pure species.

RESUMEN: El establecimiento de pautas para la gestión responsable en los sistemas de producción de peces requiere el conocimiento de la biología básica de los peces a criar. El objetivo de este trabajo fue determinar la capacidad reproductiva de los híbridos producidos por el cruce de hembras de Pseudoplatystoma metaense con machos de Leiarius marmoratus. Los machos presentaron un testículo espermatogonial digital no restringido que contiene dígitos caudales que producen glucoproteínas que no forman una vesícula seminal. Fue posible encontrar esperma libre en la luz de los túbulos y en los conductos. Se encontró que el ovario de las hembras era sacular y sincrónico con al menos tres grupos de ovocitos. En el primer año de vida, solo se encontraron oogonia hasta ovocitos previtelogénicos (alvéolo cortical de 284,9 ± 35,7 mm de diámetro). Después del segundo año, se observaron ovocitos vitelogénicos de 730 ± 3,78 mm de diámetro. Los eventos de desarrollo gonadal de los híbridos indican que son animales sincrónicos gonocóricos. La maduración alcanza su punto máximo en la temporada de aguas altas, superponiéndose con las especies parentales. Por lo tanto, el escape de híbridos de cultivos de peces a los ríos puede aumentar el riesgo de cruces, introgresión genética o disminución de la capacidad reproductiva de las especies puras.

Macrophage-induced reactive oxygen species promote myometrial contraction and labor-associated mechanisms†.

At labor, the myometrium is infiltrated by a massive influx of macrophages that secrete high levels of pro-inflammatory cytokines inducing the expression of specific labor-associated markers. However, the interactions between myocytes and macrophages and the role of macrophages in the myometrium at labor remain to be elucidated. In this work, we studied the role of myometrium-infiltrated macrophages and their interaction with myocytes in lipopolysaccharide-induced preterm labor. A co-culture model of human primary myometrial cells and macrophages was developed and validated. Collagen lattices were used to evaluate myocyte contraction. Differentiation steps were assessed by (i) phalloidin and vinculin staining for cytoskeleton reorganization, (ii) gap junction protein alpha 1 expression and scrape loading/dye transfer with Lucifer Yellow for gap junction intercellular communication, and (iii) calcium imaging for cell excitability. We demonstrated that macrophages favored lipopolysaccharide-induced contraction and early differentiation of myometrial cells. Transwell assays showed that previous activation of macrophages by lipopolysaccharide was essential for this differentiation and that macrophage/myocyte interactions involved macrophage release of reactive oxygen species (ROS). The effects of macrophage-released ROS in myometrial cell transactivation were mimicked by H2O2, suggesting that superoxide anion is a major intermediate messenger in macrophage/myocyte crosstalk during labor. These novel findings provide the foundation for innovative approaches to managing preterm labor, specifically the use of antioxidants to inhibit the initial stages of labor before the contractile phenotype has been acquired. In addition, the co-culture model developed by our team could be used in future research to decipher pathophysiological signaling pathways or screen/develop new tocolytics.

Characterization of LuWRKY36, a flax transcription factor promoting secoisolariciresinol biosynthesis in response to Fusarium oxysporum elicitors in Linum usitatissimum L. hairy roots.

MAIN CONCLUSION: The involvement of a WRKY transcription factor in the regulation of lignan biosynthesis in flax using a hairy root system is described. Secoisolariciresinol is the main flax lignan synthesized by action of LuPLR1 (pinoresinol-lariciresinol reductase 1). LuPLR1 gene promoter deletion experiments have revealed a promoter region containing W boxes potentially responsible for the response to Fusarium oxysporum. W boxes are bound by WRKY transcription factors that play a role in the response to stress. A candidate WRKY transcription factor, LuWRKY36, was isolated from both abscisic acid and Fusarium elicitor-treated flax cell cDNA libraries. This transcription factors contains two WRKY DNA-binding domains and is a homolog of AtWRKY33. Different approaches confirmed LuWRKY36 binding to a W box located in the LuPLR1 promoter occurring through a unique direct interaction mediated by its N-terminal WRKY domain. Our results propose that the positive regulator action of LuWRKY36 on the LuPLR1 gene regulation and lignan biosynthesis in response to biotic stress is positively mediated by abscisic acid and inhibited by ethylene. Additionally, we demonstrate a differential Fusarium elicitor response in susceptible and resistant flax cultivars, seen as a faster and stronger LuPLR1 gene expression response accompanied with higher secoisolariciresinol accumulation in HR of the resistant cultivar.

The control exerted by ABA on lignan biosynthesis in flax (Linum usitatissimum L.) is modulated by a Ca2+ signal transduction involving the calmodulin-like LuCML15b.

The LuPLR1 gene encodes a pinoresinol lariciresinol reductase responsible for the biosynthesis of (+)-secoisolariciresinol, a cancer chemopreventive lignan, highly accumulated in the seedcoat of flax (Linum usitatissimum L.). Abscisic acid (ABA) plays a key role in the regulation of LuPLR1 gene expression and lignan accumulation in both seeds and cell suspensions, which require two cis-acting elements (ABRE and MYB2) for this regulation. Ca2+ is a universal secondary messenger involved in a wide range of physiological processes including ABA signaling. Therefore, Ca2+ may be involved as a mediator of LuPLR1 gene expression and lignan biosynthesis regulation exerted by ABA. To test the potential implication of Ca2+ signaling, a pharmacological approach was conducted using both flax cell suspensions and maturing seed systems coupled with a ß-glucuronidase reporter gene experiment, RT-qPCR analysis, lignan quantification as well as Ca2+ fluorescence imaging. Exogenous ABA application results in an increase in the intracellular Ca2+ cytosolic concentration, originating mainly from the extracellular medium. Promoter-reporter deletion experiments suggest that the ABRE and MYB2 cis-acting elements of the LuPLR1 gene promoter functioned as Ca2+-sensitive sequences involved in the ABA-mediated regulation. The use of specific inhibitors pointed the crucial roles of the Ca2+ sensors calmodulin-like proteins and Ca2+-dependent protein kinases in this regulation. This regulation appeared conserved in the two different studied systems, i.e. cell suspensions and maturing seeds. A calmodulin-like, LuCML15b, identified from gene network analysis is proposed as a key player involved in this signal transduction since RNAi experiments provided direct evidences of this role. Taken together, these results provide new information on the regulation of plant defense and human health-promoting compounds, which could be used to optimize their production.

Preparation of a fish embryo for micromanipulation: staging of development, removal of the chorion and traceability of PGCs in Prochilodus lineatus.

The transplantation of primordial germ cells (PGCs) is a valuable tool for gene-banking and reconstitution by means of a germline chimera. For this technology, studies regarding developmental stages and traceability of PGCs are necessary. The objective of this study was to develop a micromanipulation procedure for the future establishment of cryobanks of PGCs in migratory characins. Incubation temperatures were evaluated at 22 ° C, 26 ° C, and 30 ° C in order to synchronize developmental stages. The highest hatching rates and the lowest abnormality rate arose at 26° C, which was considered to be the best incubation temperature. Enzymatic removal of the chorion was determined to be best using 0.05% pronase, in which the embryos presented better survival rates. In order to visualize PGCs in vivo, artificial GFP-nos1 3'UTR mRNA was injected and the migration route was observed in vivo as PGCs were visualized firstly at the segmentation stage (6 to 13 somites). The number of GFP positive cells ranged from 8 to 20 per embryo (mean of 13.8; n = 5). After hatching, GFP-positive cells increased to 14 to 27 embryos (mean of 19.8; n = 5). Visualization of the GFP-positive cells was possible at 10 days post hatching, and at this stage, the cells were positioned in the yolk extension region. This is the first report on PGC visualization in vivo in Neotropical fish; the obtained data provide information on the identification and migration of PGCs. The information presented in this work brings new insights in gene banking in Neotropical species and subsequent reconstitution through a germinal germline chimera.

Determinación, diferenciación sexual y pubertad en peces / Sex Determination, Sex Differentiation and Puberty in Fish / Determinação, Diferenciação Sexual e Puberada em Peixes

Resumen En piscicultura es de gran utilidad conocer los procesos de determinación sexual y los mecanismos que permiten la diferenciación sexual, así como el momento en el cual se alcanza la madurez y los individuos son capaces de reproducirse. Estos tres procesos son muy flexibles en peces y no solo dependen de diferentes cascadas de genes, si no que algunos factores ambientales son condicionantes o desencadenantes de ellos. En consecuencia, esta revisión pretende dar una mirada a los últimos avances sobre el tema, particularmente las interpretaciones científicas de los eventos de maduración sexual y sus mecanismos reguladores.

Abstract In fish farming is useful to know the processes of sex determination and mechanisms that allow differentiation and the time in which maturation is reached and specimens are able to reproduce, these three processes are very flexible in specimens and not just rely different cascades of genes if some environmental factors are conditions or triggers them. So this review aims to take a look at the latest articles on the subject, scientific interpretations of the events of sexual maturation and their regulatory mechanisms, and review the management of his control.

Resumo Na piscicultura, é muito útil conhecer os processos de determinação sexual e os mecanismos que permitem a diferenciação sexual, assim como o momento em que a maturidade é alcançada e os indivíduos são capazes de se reproduzir. Esses três processos são muito flexíveis em peixes e não dependem apenas de diferentes cascatas de genes, mas alguns fatores ambientais condicionam ou acionam esses genes. Consequentemente, esta revisão pretende dar uma olhada nos últimos desenvolvimentos sobre o assunto, particularmente as interpretações científicas dos eventos de maturação sexual e seus mecanismos regulatórios.

zHSF1 modulates zper2 expression in zebrafish embryos.

HSF1 is a transcription factor that plays a key role in circadian resetting by temperature. We have used zebrafish embryos to decipher the roles of zHsf1, heat and light on zper2 transcription in vivo. Our results show that heat shock (HS) stimulated zper2 expression in the dark but has no cumulative effect combined with light. After light exposition, zper2 expression was 2.7 fold increased threefold in the hsf1-morphants in comparison to control embryos. Our results show that zHsf1 plays a positive role in HS-driven expression of zper2 in the dark but seems to act as an attenuator in the presence light.

Evaluación de un programa de detección temprana de cáncer cervicouterino en Colombia / Evaluation of A Cervical Cancer Early Detection Program In Colombia

Resumen Objetivo: Evaluar el proceso de atención del programa de detección temprana de cáncer cervicouterino enfocado en cobertura de tamización, oportunidad de atención y adherencia en el direccionamiento del diagnóstico y tratamiento en seis municipios de Colombia. Métodos: Estudio observacional descriptivo de corte transversal a partir del análisis de fuentes secundarias de información del programa de detección temprana de cáncer cervicouterino de una aseguradora en salud de personas afiliadas con vinculación laboral y capacidad de pago. El procesamiento y análisis de los datos fueron realizados mediante el programa estadístico SPSS® versión 22 con la identificación de frecuencias absolutas, relativas, medidas de tendencia central y variabilidad. Resultados: Para el 2014, se tomaron 28.442 citologías cervicales, el 2,3% fueron resultados con algún tipo de anormalidad. Se evidenció cobertura anual de tamización del 28,2% (0,0% -280,6%), oportunidad de atención; entre toma y entrega de resultado: 15 días (4 -118), entre el resultado y el diagnóstico: 18 días (2 - 294) y entre el diagnóstico y tratamiento: 49,5 días (2 -240). Para adherencia, el 81,6% evidencia cumplimiento del direccionamiento al diagnóstico a partir del resultado citológico y el 62,6% evidencia cumplimiento en el direccionamiento del diagnóstico al tratamiento. Conclusiones: El programa de detección temprana de cáncer cervicouterino presenta coberturas bajas de tamización, falencias en el cumplimiento de la oportunidad en la atención y de la adherencia en el direccionamiento del diagnóstico y del tratamiento.

Abstract Objective: To evaluate the care process of the Cervical Cancer Early Detection program focused on screening coverage, access to care, and adherence when addressing the diagnosis and treatment, in six towns of Colombia. Methods: A cross-sectional, descriptive, observational study conducted using the analysis of secondary information sources from the Cervical Cancer Early Detection program of an occupational health insurer of persons employed, and with the ability to pay. Data processing and analysis were performed using the statistical program SPSS® version 22 with the identification of absolute and relative frequencies, central tendency, and variability measurements. Results: In the year 2014, of the 28,442 PAP smear tests taken, 2.3% were found to have some kind of anomaly. An annual screening coverage of 28.2% (0.0% - 280.6%) was observed. The access to care between the date of the sample and the delivery of the result was 15 days (4 - 118), between the result of the test and the diagnosis: 18 days (2 - 294), and 49.5 days (2 - 240) between the diagnosis and treatment. For adherence, 81.6% showed adherence to the diagnosis based on the cytological result, and 62.6% showed compliance in the diagnosis-treatment approach. Conclusions: The cervical cancer early detection program has a low screening coverage, with failures in complying with timely care and adherence in addressing diagnosis and treatment.

Efecto del protocolo de inducción hormonal en la espermiación y las características seminales de yaque ( Leiarius marmoratus ) / Effect of hormone induction protocol in the spermiation and seminal characteristics of yaque (Leiarius marmoratus) / Efeito do protocolo de indução hormonal sobre a spermiação e as características do sêmen do yaque (Leiarius marmoratus)

Resumen El yaque Leiarius marmoratus, es una especie de siluriforme nativo con gran potencial para la diversificación de la acuicultura colombiana; sin embargo, la espermiación en cautiverio aún presenta dificultades debido al bajo volumen seminal liberado con los protocolos de inducción tradicionales. En consecuencia, el objetivo fue evaluar los efectos de cinco protocolos de inducción hormonal, basados en Extracto de Hipófisis de Carpa (EHC), sGnRHa + domperidona (OVAPRIM®) y Gonadotropina Coriónica Humana (GHC) (FERTIVET®), sobre la calidad y volumen seminal de la especie. Fueron seleccionados 18 machos sexualmente maduros de 54,4 ± 2,2 cm de longitud total y 1,6 ± 0,1 kg peso corporal. Se evaluaron 5 tratamientos, así: T1: 0,25 ml/kg (0 h) y 1 ml/kg de OVAPRIM® (12 h); T2: 1 mg/kg de EHC (0 h) y 3 mg/kg de EHC (12 h); T3: dosis única de 200 UI/kg de GCH; T4: 3 mg/kg de EHC (0 h) y 200 UI/Kg de GHC (8h); T5: 1 ml/kg de OVAPRIM® (0h) y 200 UI/kg de GHC (8h); Control: 1 ml de suero fisiológico. El semen se colecto 6 h después de aplicada la última dosis hormonal. El T1 y T5 mostraron un volumen mayor (p<0,05) (5,4 ± 1,1; 4,8 ± 1,2 ml) cuando comparados con T2 (3,1 ± 1ml), T3 (2,3 ml) T4 (2,4 ± 1,1 ml) y el control (0,2 ml). En cuanto a la movilidad y tiempo de activación, T1 y T5 presentaron los mejores resultados (90 ± 0% y 54,2 ± 7 s; 90 ± 0% y 52,9 ± 3,8 s, respectivamente). La concentración espermática fue mayor (p<0,05) en T2 y T5 (1,275 ± 322; y 1,261 ± 225 sptz x 103/µl) comparadas con los otros tratamientos. Se concluye que la inducción hormonal con OVAPRIM® combinada con HCG, son efectivas para la producción seminal en la especie.

Abstract The yaque Leiarius marmoratus is specie of native silurid, with great potential for the diversification of the colombian aquaculture; however, the spermiation in captive yet presents difficulties due to the low seminal volume released with traditional induction protocols. In consequence, the aim of this study was to evaluate the effects of five protocols of hormonal induction based on carp pituitary extract (EHC), sGnRHa + domperidone (OVAPRIM®) and human chorionic gonadotropin (GHC) (FERTIVET®) on seminal quality and volume of the specie. Were selected 18 sexually mature males of 54,4 ± 2,2 cm in total length and 1,6 ± 0,1 kg body weight. 5 treatments were evaluated so: T1 : 0,25 ml/kg (0 h) and 1 ml/kg of OVAPRIM® (12 h ); T2: 1 mg/kg of EHC (0 h) and 3 mg/kg of EHC (12 h) ; T3 : single dose of 200 UI/kg HCG; T4: 3 mg/kg of EHC (0 h) and 200 UI /kg GHC (8h) ; T5: 1 ml/kg of OVAPRIM® (0h) and 200 U.I / kg of GHC (8h) ; Control: 1 ml of saline solution. Semen was obtained 6 hours after the last dose hormone. T1 and T5 showed a larger volume (p<0.05) (5,4 ± 1,1, 4,8 ± 1,2 ml) compared to T2 (3,1 ± 1ml), T3 (2,3 ml) T4 (2,4 ± 1.1 ml) and control (0.2 ml). Regarding motility and activation time, T1 and T5 had the best results (90 ± 0 % and 54,2 ± 7 s; 90 ± 0 % and 52,9 ± 3,8 s, respectively). The sperm concentration was higher in (p<0,05) T2 and T5 (1,275 ± 322, ± 225 and 1,261 sptz x 103/ul) compared with the other treatments. In conclusion, hormonal induction with OVAPRIM® combined with HCG, are effective for the seminal production in the specie.

Resumo O yaque Leiarius marmoratus, é uma espécie siluriforme nativa com grande potencial para a diversificação da aquicultura colombiana; No entanto, a spermiação em cativeiro ainda apresenta dificuldades devido ao baixo volume seminal liberado com protocolos de indução tradicionais. Por conseguinte, o objetivo foi avaliar os efeitos de protocolos de indução de cinco hormonais base de extrato de hipófise de carpa (EHC), sGnRHa + domperidona (Ovaprim®) e Gonadotropina coriónica humana (GHC) (FERTIVET®) sobre a qualidade e volume seminal das espécies. Foram Selecionados 18 machos sexualmente maduros de 54,4 ± 2,2 cm de comprimento total e 1,6 ± 0,1 kg de peso corporal. Foram avaliados cinco tratamentos: T1: 0,25 ml / kg (0 h) e 1 ml / kg de OVAPRIM® (12 h); T2: 1 mg / kg de EHC (0 h) e 3 mg / kg de EHC (12 h); T3: dose única de 200 UI / kg de HCG; T4: 3 mg / kg de EHC (0 h) e 200 UI / kg de GHC (8h); T5: 1 ml / kg de OVAPRIM® (0h) e 200 UI / kg de GHC (8h); Controle: 1 ml de solução salina fisiológica. O sêmen foi coletado 6 h após a última dose hormonal foi aplicada. T1 e T5 mostraram um volume maior (p <0,05) (5,4 ± 1,1, 4,8 ± 1,2 ml) quando comparado com T2 (3,1 ± 1 ml), T3 (2,3 ml) T4 (2,4 ± 1,1 ml) e controle (0,2 ml). Em relação à mobilidade e ao tempo de ativação, T1 e T5 apresentaram os melhores resultados (90 ± 0% e 54,2 ± 7 s, 90 ± 0% e 52,9 ± 3,8 s, respectivamente). A concentração de esperma foi maior (p <0,05) em T2 e T5 (1275 ± 322 e 1261 ± 225 sptz x 103 / µl) em comparação com os outros tratamentos. Conclui-se que a indução hormonal com OVAPRIM® combinada com HCG, é efetiva para a produção seminal nas espécies.

Functional characterization of the pinoresinol-lariciresinol reductase-2 gene reveals its roles in yatein biosynthesis and flax defense response.

MAIN CONCLUSION: This study provides new insights into the biosynthesis regulation and in planta function of the lignan yatein in flax leaves. Pinoresinol-lariciresinol reductases (PLR) catalyze the conversion of pinoresinol into secoisolariciresinol (SECO) in lignan biosynthesis. Several lignans are accumulated in high concentrations, such as SECO accumulated as secoisolariciresinol diglucoside (SDG) in seeds and yatein in aerial parts, in the flax plant (Linum usitatissimum L.) from which two PLR enzymes of opposite enantioselectivity have been isolated. While LuPLR1 catalyzes the biosynthesis of (+)-SECO leading to (+)-SDG in seeds, the role(s) of the second PLR (LuPLR2) is not completely elucidated. This study provides new insights into the in planta regulation and function of the lignan yatein in flax leaves: its biosynthesis relies on a different PLR with opposite stereospecificity but also on a distinct expression regulation. RNAi technology provided evidence for the in vivo involvement of the LuPLR2 gene in the biosynthesis of (-)-yatein accumulated in flax leaves. LuPLR2 expression in different tissues and in response to stress was studied by RT-qPCR and promoter-reporter transgenesis showing that the spatio-temporal expression of the LuPLR2 gene in leaves perfectly matches the (-)-yatein accumulation and that LuPLR2 expression and yatein production are increased by methyl jasmonate and wounding. A promoter deletion approach yielded putative regulatory elements. This expression pattern in relation to a possible role for this lignan in flax defense is discussed.

The impact of tumor nitric oxide production on VEGFA expression and tumor growth in a zebrafish rat glioma xenograft model.

To investigate the effect of nitric oxide on tumor development, we established a rat tumor xenograft model in zebrafish embryos. The injected tumor cells formed masses in which nitric oxide production could be detected by the use of the cell-permeant DAF-FM-DA (diaminofluorophore 4-amino-5-methylamino-2'-7'-difluorofluorescein diacetate) and DAR-4M-AM (diaminorhodamine-4M). This method revealed that nitric oxide production could be co-localized with the tumor xenograft in 46% of the embryos. In 85% of these embryos, tumors were vascularized and blood vessels were observed on day 4 post injection. Furthermore, we demonstrated by qRT-PCR that the transplanted glioma cells highly expressed Nos2, Vegfa and Cyclin D1 mRNA. In the xenografted embryos we also found increased zebrafish vegfa expression. Glioma and zebrafish derived Vegfa and tumor Cyclin D1 expression could be down regulated by the nitric oxide scavenger 2-(4-Carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide or CPTIO. We conclude that even if there is a heterogeneous nitric oxide production by the xenografted glioma cells that impacts Vegfa and Cyclin D1 expression levels, our results suggest that reduction of nitric oxide levels by nitric oxide scavenging could be an efficient approach to treat glioma.

Structure-function analysis of peroxisomal ATP-binding cassette transporters using chimeric dimers.

ABCD1 and ABCD2 are two closely related ATP-binding cassette half-transporters predicted to homodimerize and form peroxisomal importers for fatty acyl-CoAs. Available evidence has shown that ABCD1 and ABCD2 display a distinct but overlapping substrate specificity, although much remains to be learned in this respect as well as in their capability to form functional heterodimers. Using a cell model expressing an ABCD2-EGFP fusion protein, we first demonstrated by proximity ligation assay and co-immunoprecipitation assay that ABCD1 interacts with ABCD2. Next, we tested in the pxa1/pxa2Δ yeast mutant the functionality of ABCD1/ABCD2 dimers by expressing chimeric proteins mimicking homo- or heterodimers. For further structure-function analysis of ABCD1/ABCD2 dimers, we expressed chimeric dimers fused to enhanced GFP in human skin fibroblasts of X-linked adrenoleukodystrophy patients. These cells are devoid of ABCD1 and accumulate very long-chain fatty acids (C26:0 and C26:1). We checked that the chimeric proteins were correctly expressed and targeted to the peroxisomes. Very long-chain fatty acid levels were partially restored in transfected X-linked adrenoleukodystrophy fibroblasts regardless of the chimeric construct used, thus demonstrating functionality of both homo- and heterodimers. Interestingly, the level of C24:6 n-3, the immediate precursor of docosahexaenoic acid, was decreased in cells expressing chimeric proteins containing at least one ABCD2 moiety. Our data demonstrate for the first time that both homo- and heterodimers of ABCD1 and ABCD2 are functionally active. Interestingly, the role of ABCD2 (in homo- and heterodimeric forms) in the metabolism of polyunsaturated fatty acids is clearly evidenced, and the chimeric dimers provide a novel tool to study substrate specificity of peroxisomal ATP-binding cassette transporters.

Biological activities of Schottenol and Spinasterol, two natural phytosterols present in argan oil and in cactus pear seed oil, on murine miroglial BV2 cells.

The objective of this study was to evaluate the biological activities of the major phytosterols present in argan oil (AO) and in cactus seed oil (CSO) in BV2 microglial cells. Accordingly, we first determined the sterol composition of AO and CSO, showing the presence of Schottenol and Spinasterol as major sterols in AO. While in CSO, in addition to these two sterols, we found mainly another sterol, the Sitosterol. The chemical synthesis of Schottenol and Spinasterol was performed. Our results showed that these two phytosterols, as well as sterol extracts from AO or CSO, are not toxic to microglial BV2 cells. However, treatments by these phytosterols impact the mitochondrial membrane potential. Furthermore, both Schottenol and Spinasterol can modulate the gene expression of two nuclear receptors, liver X receptor (LXR)-α and LXRß, their target genes ABCA1 and ABCG1. Nonetheless, only Schottenol exhibited a differential activation vis-à-vis the nuclear receptor LXRß. Thus Schottenol and Spinasterol can be considered as new LXR agonists, which may play protective roles by the modulation of cholesterol metabolism.