Effects of calcitriol upon TGF-ßs and their receptors in trophoblast cells.

Calcitriol levels increase during pregnancy, contributing to the hormonal and immunological balance, but its deficiency has been associated with problems during this period. Meanwhile, transforming growth factors-ß (TGF-ßs) play an important role in the maintenance of fetal-maternal immune tolerance; however, exacerbated concentrations of this growth factor are associated with complicated pregnancies. Therefore, we studied the effects of calcitriol on TGF-ßs and their receptors in trophoblast cells. Term placentas from uncomplicated pregnancies after cesarean sections were used for cell cultures. Basal gene expression and the effect of calcitriol upon TGF-ß1, TGF-ß2, TGF-ß3, and their receptors TGF-ßR1 and TGF-ßR2 were assessed using real-time PCR from trophoblast cells. The presence of TGF-ß1, 2, 3, and TGF-ßR1 were evaluated by immunofluorescence, and the protein abundance and secretion of TGF-ß1 were assessed by Western blot and ELISA, respectively. Basal gene expression of TGF-ß1 in trophoblast from term placentas was higher than TGF-ß2 and TGF-ß3, while TGF-ßR2 was higher than TGF-ßR1. The presence and cellular localization of TGF-ß1, 2, 3, and TGF-ßR1 were detected in the cytoplasm of syncytiotrophoblast, with TGF-ß1 showing the highest intensity. Calcitriol significantly inhibited gene expression of TGF-ß1, TGF-ß2, and TGF-ßR1. Likewise, calcitriol decreased the secretion and abundance of TGF-ß1. In conclusion, results indicate that calcitriol is a regulator of TGF-ßs in cultured trophoblast cells from term placentas and therefore may be an important player in the development of healthy pregnancies.

Polycystic ovarian syndrome: signs and feedback effects of hyperandrogenism and insulin resistance.

Polycystic ovary syndrome (PCOS) is a disease whose diagnosis is based on the detection of hyperandrogenism (HA) and ovulatory dysfunction. Women with PCOS frequently develop insulin resistance (IR), which generates a metabolic condition that involves a decrease in the action of insulin at the cellular level and is linked to compensatory hyperinsulinemia (HI). In PCOS, the ovary remains sensitive to the action of insulin. Additionally, it has been observed that the main effect of insulin in the ovary is the stimulation of androgen synthesis, resulting in HA, one of the fundamental characteristics of the PCOS. In this sense, the excess of androgens favors the development of IR, thus perpetuating the cycle of IR-HI-HA, and therefore PCOS. Moreover, mitochondrial dysfunction is present in PCOS patients and is a common feature in both IR and HA. This review places electron transfer as a key element in HA and IR development, with emphasis on the relationship between androgen biosynthesis and mitochondrial function. Indeed, metformin has been involved in repair mitochondrial dysfunction, decrease of oxidative stress, reduction of androgens levels and the enhancing of insulin sensitivity. Therefore, we propose that treatment with metformin could decrease HI and consequently HA, restoring, at least in part, the metabolic and hormonal disorders of PCOS.

Regulation of anti-tumorigenic pathways by the combinatory treatment of calcitriol and TGF-ß in PC-3 and DU145 cells.

Calcitriol and transforming growth factors beta (TGF-ß) are involved in several biological pathways such as cell proliferation, differentiation, migration and invasion. Their cellular effects could be similar or opposite depending on the genetic target, cell type and context. Despite the reported association of calcitriol deficiency and disruption of the TGF-ß pathway in prostate cancer and the well-known independent effects of calcitriol and TGF-ßs on cancer cells, there is limited information regarding the cellular effects of calcitriol and TGF-ß in combination. In this study, we in vitro analyze the combinatory effects of calcitriol and TGF-ß on cell growth and apoptosis using PC-3 and DU145 human prostate cancer cell lines. Using high-throughput microarray profiling of PC-3 cells upon independent and combinatory treatments, we identified distinct transcriptional landscapes of each intervention, with a higher effect established by the combinatorial treatment, following by TGF-ß1 and later by calcitriol. A set of genes and enriched pathways converge among the treatments, mainly between the combinatory scheme and TGF-ß1, but the majority were treatment-specific. Of note, CYP24A1, IGFBP3, CDKN1A, NOX4 and UBE2D3 were significantly up-regulated upon the combinatorial treatment whereas CCNA1, members of the CT45A and APOBEC3 family were down-regulated. By public RNA signatures, we were able to confirm the regulation by the co-treatment over cell proliferation and cell cycle. We finally investigated the possible clinical impact of genes modulated by the combinatorial treatment using benchmark prostate cancer data. This comprehensive analysis reveals that the combinatory treatment impairs cell growth without affecting apoptosis and their combinatory actions might synergize and improved their individual effects to reprogram prostate cancer signaling.

An organotin indomethacin derivative inhibits cancer cell proliferation and synergizes the antiproliferative effects of lapatinib in breast cancer cells.

It is known that an inflammatory condition in different types of cancer provides a sustained microenvironment that favors tumor growth, invasion, and metastasis. Non-steroidal anti-inflammatory drugs such as indomethacin have demonstrated chemo-preventive, anti-proliferative and cytotoxic effects in a variety of tumors. The aim of this study was to investigate the effects of an organotin indomethacin derivative (OID) on the proliferation of breast and prostate cancer cell lines and the possible mechanisms of action of this compound. Different cancer cell lines were treated in the presence of OID and cell proliferation was measured by quantification of the DNA content, changes in the cell cycle profile and the activation of caspase 3 were evaluated by flow cytometry, interleukin 6 (IL-6) gene expression was evaluated by qPCR and protein expression was analyzed by ELISA and Western blot assays. OID inhibited the cell proliferation of a panel of cancer cell lines in a concentration-dependent manner. Moreover, the addition of OID to lapatinib treatment, targeted therapy for breast cancer, significantly enhanced its antiproliferative response. The effects on cell proliferation of these compounds involved, among others, the induction of apoptosis, the downregulation of IL-6 and a decrease of the MAPK activation pathway. Our results suggest that the use of OID alone or in combination with tyrosine kinase inhibitors could be considered as adjuvants in the treatment of cancer.

Multicore optical fiber shape sensors suitable for use under gamma radiation.

We have designed and implemented a fiber optic shape sensor for high-energy ionizing environments based on multicore optical fibers. We inscribed two fiber Bragg gratings arrays in a seven-core optical fiber. One of the arrays has been inscribed in a hydrogen-loaded fiber and the other one in an unloaded fiber in order to have two samples with very different radiation sensitivity. The two samples were coiled in a metallic circular structure and were exposed to gamma radiation. We have analyzed the permanent radiation effects. The radiation-induced Bragg wavelength shift (RI-BWS) in the hydrogen-loaded fiber is near ten times higher than the one observed for the unloaded fiber, with a maximum wavelength shift of 415 pm. However, the use of the multiple cores permits to make these sensors immune to RI-BWS obtaining a similar curvature error in both samples of approximately 1 cm without modifying the composition of the fiber, pre-irradiation or thermal treatment.

Isocitrate dehydrogenase type 2 (IDH2) is part of a multiprotein complex for placental steroidogenesis.

BACKGROUND: Human syncytiotrophoblast mitochondria require the activity of the isocitrate dehydrogenase type 2 (IDH2) to obtain reduced coenzymes for progesterone (P4) synthesis. Data from the literature indicate that mitochondrial steroidogenic contact sites transform efficiently cholesterol into P4. In this research, we identified the IDH2 as a member of the steroidogenic contact site and analyzed the steroidogenic role of its activity. METHOD: Human syncytiotrophoblast mitochondria were isolated by differential centrifugation, and steroidogenic contact sites were obtained by osmotic shock and sucrose gradient ultracentrifugation. In-gel native activity assay, mass spectroscopy, and western blot were used to identify the association of proteins and their activities. P4 was determined by immunofluorescence. RESULTS: The IDH2 was mainly identified in steroidogenic contact sites, and its activity was associated with a complex of proteins with an apparent molecular mass of ~590 kDa. Mass spectroscopy showed many groups of proteins with several metabolic functions, including steroidogenesis and ATP synthesis. The IDH2 activity was coupled to P4 synthesis since in the presence of Ca2+ or Na2SeO3, inhibitors of the IDH2, the P4 production decreased. CONCLUSIONS: The human syncytiotrophoblast mitochondria build contact sites for steroidogenesis. The IDH2, a non-membrane protein, supplies the NADPH required for the synthesis of P4 in a complex (steroidosome) that associate the proteins required to transform efficiently cholesterol into P4, which is necessary in pregnancy to maintain the relationship between mother and fetus. GENERAL SIGNIFICANCE: The IDH2 is proposed as a check point in the regulation of placental steroidogenesis.

Calcitriol Inhibits the Proliferation of Triple-Negative Breast Cancer Cells through a Mechanism Involving the Proinflammatory Cytokines IL-1ß and TNF-α.

Triple-negative breast cancer (TNBC) is one of the most aggressive tumors, with poor prognosis and high metastatic capacity. The aggressive behavior may involve inflammatory processes characterized by deregulation of molecules related to the immunological responses in which interleukin-1ß (IL-1ß) and tumor necrosis factor-α (TNF-α) are involved. It is known that calcitriol, the active vitamin D metabolite, modulates the synthesis of immunological mediators; however, its role in the regulation of IL-1ß and TNF-α in TNBC has been scarcely studied. In the present study, we showed that TNBC cell lines SUM-229PE and HCC1806 expressed vitamin D, IL-1ß, and TNF-α receptors. Moreover, calcitriol, its analogue EB1089, IL-1ß, and TNF-α inhibited cell proliferation. In addition, we showed that synthesis of both IL-1ß and TNF-α was stimulated by calcitriol and its analogue. Interestingly, the antiproliferative activity of calcitriol was significantly abrogated when the cells were treated with anti-IL-1ß receptor 1 (IL-1R1) and anti-TNF-α receptor type 1 (TNFR1) antibodies. Furthermore, the combination of calcitriol with TNF-α resulted in a greater antiproliferative effect than either agent alone, in the two TNBC cell lines and an estrogen receptor-positive cell line. In summary, this study demonstrated that calcitriol exerted its antiproliferative effects in part by inducing the synthesis of IL-1ß and TNF-α through IL-1R1 and TNFR1, respectively, in TNBC cells, highlighting immunomodulatory and antiproliferative functions of calcitriol in TNBC tumors.

Steroid hormones and pregnancy.

Pregnancy is associated with physiological adjustments in order to allow adequate growth and fetal development. In particular, steroids are necessary to maintain in balance numerous functions during gestation. Steroidogenesis in the maternal, placental and fetal compartments and the biological effects of progestins and estrogens that play a pivotal role before and during pregnancy are described. Although it is well-known that androgens are considered as substrate for estrogens biosynthesis, their biosynthesis and functionality in placental and other tissues have been questioned. As compared with healthy pregnancy, steroid hormones levels have been found altered in complicated pregnancies and hormonal treatments have been used is some pathologies. Therefore, the aim of this work was to review the biosynthesis, function and regulation of progestins, androgens and estrogens during gestation. Furthermore, steroid hormones concentrations during healthy and complicated pregnancy as well hormonal therapies for the prevention of miscarriages and preterm deliveries are discussed in the present review.

Selección de hospital destino para el traslado de urgencia de pacientes / Selecting the Destination Hospital to Carry Emergency Patients / Escolha de hospital alvo para traslado de urgência de pacientes

Resumen El tiempo total de atención prehospitalaria (APH) es el tiempo que transcurre desde que ingresa la llamada al operador hasta que la ambulancia queda disponible para atender otra emergencia. Esta investigación pretende demostrar que la selección del hospital destino afecta de manera significativa el tiempo total de APH, lo que influye en la supervivencia del paciente que es trasladado y en el tiempo de liberación del recurso (ambulancias). En consecuencia, se propone una técnica de selección de hospital destino que incluye dimensiones relacionadas con el paciente (diagnóstico, especialidad y asegurador) y el hospital (ocupación y cercanía). Se evalúa su desempeño por medio de una simulación de eventos discretos y se concluye que la técnica propuesta obtiene un mejor tiempo de APH en el 73% de los casos estudiados, con una reducción media entre 40 y 80 minutos, en comparación con la técnica más comúnmente usada (selección hospital más cercano).

Abstract Total time of prehospital care (PHC) is the time elapsing from the inbound call up to the moment when the ambulance is available for serve in another emergency event. This research aims to show that selecting the destination hospital impacts significantly the total PHC time, which influences the survival of the patient being transported as well as the time to make the resource available again (the ambulance). Consequently, a technique for selecting the destination hospital is proposed herein including some dimensions related both to the patient (diagnosis, specialty and insurance company) and to the hospital (occupancy and closeness). The performance was evaluated based on a simulation of discrete events. It is concluded that the proposed technique provides a better PHC time in 73% of the studied cases, with a mean decrease between 40 and 80 minutes as compared to the most commonly used technique (selecting the closest hospital).

Resumo O tempo total de atendimento pré-hospitalar (APH) é o tempo decorrente desde que a ligação for feita para o telefonista até a ambulância se disponibilizar para atender outra emergência. Esta pesquisa visa demostrar que a escolha do hospital alvo afeta significativamente o tempo total de APH, o que influi na sobrevida do paciente trasladado e no tempo de liberação do recurso (ambulâncias). Consequentemente, propõe-se uma técnica de escolha de hospital alvo que inclui dimensões relacionadas com paciente (diagnóstico, especialidade e assegurador) e hospital (ocupação e proximidade). Avalia-se o desempenho por meio de simulação de eventos discretos e conclui-se que a técnica proposta obtém melhor tempo de APH em 73% dos casos estudados com redução media entre 40 e 80 minutos, em comparação com a técnica mais comumente usada (escolha hospital mais próximo).

A single preovulatory administration of ulipristal acetate affects the decidualization process of the human endometrium during the receptive period of the menstrual cycle.

In order to get further information on the effects of ulipristal acetate (UPA) upon the process of decidualization of endometrium, a functional analysis of the differentially expressed genes in endometrium (DEG) from UPA treated-versus control-cycles of normal ovulatory women was performed. A list of 1183 endometrial DEG, from a previously published study by our group, was submitted to gene ontology, gene enrichment and ingenuity pathway analyses (IPA). This functional analysis showed that decidualization was a biological process overrepresented. Gene set enrichment analysis identified LIF, PRL, IL15 and STAT3 among the most down-regulated genes within the JAK STAT canonical pathway. IPA showed that decidualization of uterus was a bio-function predicted as inhibited by UPA. The results demonstrated that this selective progesterone receptor modulator, when administered during the periovulatory phase of the menstrual cycle, may affect the molecular mechanisms leading to endometrial decidualization in response to progesterone during the period of maximum embryo receptivity.

Strategies for the quality assessment of the health care service providers in the treatment of Gastric Cancer in Colombia.

BACKGROUND: While, at its inception in 1993, the health care system in Colombia was publicized as a paradigm to be copied across the developing world, numerous problems in its implementation have led to, what is now, an inefficient and crisis-ridden health system. Furthermore, as a result of inappropriate tools to measure the quality of the health service providers, several corruption scandals have arisen in the country. This study attempts to tackle this situation by proposing a strategy for the quality assessment of the health service providers (Entidades Promotoras de Salud, EPS) in the Colombian health system. In particular, as a case study, the quality of the treatment of stomach cancer is analyzed. METHODS: The study uses two complementary techniques to address the problem. These techniques are applied based on data of the treatment of gastric cancer collected on a nation-wide scale by the Colombian Ministry of Health and Welfare. First, Data Envelopment Analysis (DEA) and the Malmquist Index (MI) are used to establish the most efficient EPS's within the system, according to indicators such as opportunity indicators. Second, sequential clustering algorithm, related to process mining a field of data mining, is used to determine the medical history of all patients and to construct typical care pathways of the patients belonging to efficient and inefficient EPS's. Lastly, efforts are made to identify traits and differences between efficient and inefficient EPS's. RESULTS: Efficient and inefficient EPS were identified for the years 2010 and 2011. Additionally, a Malmquist Index was used to calculate the relative changes in the efficiency of the health providers. Using these efficiency rates, the typical treatment path of patients with gastric cancer was found for two EPSs: one efficient and another inefficient. Finally, the typical traits of the care pathways were established. CONCLUSIONS: Combining DEA and process mining proved to be a powerful approach understanding the problem and gaining valuable insight into the inner workings of the Colombian Health System, especially in terms of the treatment process performed by health care providers in critical illnesses such as cancer. However, no sufficiently compelling results were found to establish the contribution of such a combination to evaluate the quality in the delivery of health services.

Ulipristal acetate administration at mid-cycle changes gene expression profiling of endometrial biopsies taken during the receptive period of the human menstrual cycle.

The aim of this study was to analyze the effects of mid-cycle administration of Ulipristal acetate (UPA) on gene expression in endometrial biopsies taken during the receptive phase of the cycle. Fourteen healthy menstruating women were studied during 14 control non-treated and 12 treated cycles with a single dose of 30 mg UPA when follicle diameter reached 20 mm. Ovulation in both treated and control cycles was confirmed by serial determinations of serum LH, progesterone and vaginal ultrasound. An endometrial biopsy at day LH+7, in each cycle, was taken for RNA microarray and qPCR analysis or prepared for histological and immunohistochemistry studies. Functional analysis of differentially expressed genes showed the presence of changes compatible with a non-receptive endometrial phenotype, further confirmed by qPCR and immunohistochemistry. This study suggests the effects of UPA on endometrial receptivity, offering a plausible explanation for the higher contraceptive efficacy of this method compared to that of levonorgestrel.

Chronic moderate ethanol intake differentially regulates vitamin D hydroxylases gene expression in kidneys and xenografted breast cancer cells in female mice.

Factors affecting vitamin D metabolism may preclude anti-carcinogenic effects of its active metabolite calcitriol. Chronic ethanol consumption is an etiological factor for breast cancer that affects vitamin D metabolism; however, the mechanisms underlying this causal association have not been fully clarified. Using a murine model, we examined the effects of chronic moderate ethanol intake on tumoral and renal CYP27B1 and CYP24A1 gene expression, the enzymes involved in calcitriol synthesis and inactivation, respectively. Ethanol (5% w/v) was administered to 25-hydroxyvitamin D3-treated or control mice during one month. Afterwards, human breast cancer cells were xenografted and treatments continued another month. Ethanol intake decreased renal Cyp27b1 while increased tumoral CYP24A1 gene expression.Treatment with 25-hydroxyvitamin D3 significantly stimulated CYP27B1 in tumors of non-alcohol-drinking mice, while increased both renal and tumoral CYP24A1. Coadministration of ethanol and 25-hydroxyvitamin D3 reduced in 60% renal 25-hydroxyvitamin D3-dependent Cyp24a1 upregulation (P<0.05). We found 5 folds higher basal Cyp27b1 than Cyp24a1 gene expression in kidneys, whereas this relation was inverted in tumors, showing 5 folds more CYP24A1 than CYP27B1. Tumor expression of the calcitriol target cathelicidin increased only in 25-hydroxyvitamin D3-treated non-ethanol drinking animals (P<0.05). Mean final body weight was higher in 25-hydroxyvitamin D3 treated groups (P<0.001). Overall, these results suggest that moderate ethanol intake decreases renal and tumoral 25-hydroxyvitamin D3 bioconversion into calcitriol, while favors degradation of both vitamin D metabolites in breast cancer cells. The latter may partially explain why alcohol consumption is associated with vitamin D deficiency and increased breast cancer risk and progression.

Reglas de despacho en la programación de procedimientos quirúrgicos electivos: impacto en los indicadores de ocupación y oportunidad / Dispatching Rules in Elective Surgery Scheduling: Impact on Service Occupation and Opportunity / Regras de despacho na programação de procedimentos cirúrgicos eletivos: impacto nos indicadores de ocupação e oportunidade

Introducción: la programación de los quirófanos es el factor de mayor incidencia en el desempeño de los servicios de cirugías. Este estudio cuantificó el impacto de la variabilidad artificial creada por una programación manual de las cirugías en el Hospital Universitario Mayor - Méderi (HUM), Colombia. La hipótesis planteada es que una programación semiautomática podría: (i) reducir la variabilidad diaria del servicio de cirugías, (ii) aumentar la disponibilidad de los quirófanos y (iii) mejorar el tiempo de oportunidad por cirugía. Materiales y métodos: este estudio empleó los registros del servicio de cirugía de un mes regular. El servicio estudiado ejecuta alrededor de 35 000 cirugías al año y el proceso de programación es manual. La programación real fue comparada con las generadas a partir del empleo del algoritmo Bin Packing y las reglas de despacho Longest Processing Time (LPT) y Shortest Processing Time (SPT). Resultados: la aplicación del algoritmo con la regla LPT logró una mejora en la programación del mes estudiado: los coeficientes de variación del flujo de pacientes y ocupación diaria se redujeron (25,09% y 36,71%, respectivamente). Adicionalmente, el tiempo de oportunidad se redujo en 6,2 días y la ocupación del servicio subió un 26,22%. La programación con la regla SPT aumentó la variabilidad en el flujo de paciente en 22,7% y disminuyó la ocupación en 2,28%. Conclusiones: una programación semiautomática de las salas de cirugía en el HUM empleando la regla LPT lograría mejorar sustancialmente indicadores de variabilidad del servicio, tiempo de oportunidad y ocupación.

Introduction: The performance of a surgery service is highly impacted by its schedule. This study measured the impact of the artificial variability caused by a manual scheduling of surgeries at "Hospital Universitario Mayor - Méderi (HUM)" (Colombia). The hypotheses were that the proposed algorithm is able to (i) reduce daily service variation, (ii) increase the availability of service resources, and (iii) improve the opportunity time for each surgery. Materials and methods: The studied surgical service performs around 35 000 annual surgeries and its scheduling process is presently manually made. Actual scheduling records of a regular month were compared to schedules generated by a Bin Packing (BP) algorithm hybridized with the Longest Processing Time (LPT) and Shortest Processing Time (SPT) dispatching rules. Results: It was found that the BP algorithm with LPT rule could improve service performance, reducing the variation coefficients of patients' flow and daily service occupation by 25.09% and 36.71%, respectively. The programmed surgeries were also moved ahead 6.2 days, and the overall occupation rate increased by 26.72%. Results were not better when a SPT rule was used, boosting the variability on patient flow by 22.7% and reducing the occupation by 2.28%. Conclusions: Semiautomatic scheduling of the surgical service at the HUM, a BP algorithm with LPT rule, may substantially increase service performance in terms of service occupation and opportunity.

Introdução: A programação das salas de cirurgia é o fator de maior incidência no desempenho dos serviços de cirurgias. Este estudo quantificou o impacto da variabilidade artificial criada por uma programação manual das cirurgias no Hospital Universitário Mayor - Méderi (HUM), Colômbia. A hipótese apresentada é que uma programação semiautomática poderia: (i) reduzir a variabilidade diária do serviço de cirurgias, (ii) aumentar a disponibilidade das salas de cirurgia, e (iii) melhorar o tempo de oportunidade por cirurgia. Materiais e métodos: Este estudo empregou os registros do serviço de cirurgia de um mês regular. O serviço estudado executa cerca de 35 000 cirurgias por ano e o processo de programação é manual. A programação real foi comparada com as geradas a partir do emprego do algoritmo Bin Packing e as regras de despacho Longest Processing Time (LPT) e Shortest Processing Time (SPT). Resultados: A aplicação do algoritmo com a regra LPT conseguiu uma melhora na programação do mês estudado: os coeficientes de variação do fluxo de pacientes e ocupação diária reduziram-se (25,09% y 36,71%, respectivamente). Adicionalmente, o tempo de oportunidade reduziu-se em 6,2 dias e a ocupação do serviço subiu um 26,22%. A programação com a regra SPT aumentou a variabilidade no fluxo de paciente em 22,7% e diminuiu a ocupação em 2,28%. Conclusões: Uma programação semiautomática das salas de cirurgia no HUM empregando a regra LPT conseguiria melhorar substancialmente indicadores de variabilidade do serviço, tempo de oportunidade e ocupação.

Synthesis and biological activity of two pregnane derivatives with a triazole or imidazole ring at C-21.

Pregnane derivatives are studied as agents for the treatment of different hormone-dependent diseases. The biological importance of these steroids is based on their potential use against cancer. In this study, we report the synthesis, characterization and biological activity of two pregnane derivatives with a triazole (3ß-hydroxy-21-(1H-1,2,4-triazol-1-yl)pregna-5,16-dien-20-one; T-OH) or imidazole (3ß-hydroxy-21-(1H-imidazol-1-yl)pregna-5,16-dien-20-one; I-OH) moieties at C-21. These derivatives were synthesized from 16-dehydropregnenolone acetate. The activity on cell proliferation of the compounds was measured on three human cancer cells lines: prostate cancer (PC-3), breast cancer (MCF7) and lung cancer (SK-LU-1). The cytotoxic and antiproliferative effects of T-OH and I-OH were assessed by using SBR and XTT methods, respectively. The gene expressions were evaluated by real time PCR. In addition, results were complemented by docking studies and transactivation assays using an expression vector to progesterone and androgen receptor. Results show that the two compounds inhibited the three cell lines proliferation in a dose-dependent manner. Compound I-OH downregulated the gene expression of the cyclins D1 and E1 in PC-3 and MFC7 cells; however, effect upon Ki-67, EAG1, BIM or survivin genes was not observed. Docking studies show poor interaction with the steroid receptors. Nevertheless, the transactivation assays show a weak antagonist effect of I-OH on progesterone receptor but not androgenic or antiandrogenic actions. In conclusion, the synthesized compounds inhibited cell proliferation as well as genes key to cell cycle of PC-3 and MCF7 cell lines. Therefore, these compounds could be considered a good starting point for the development of novel therapeutic alternatives to treat cancer.

Efficacy and mechanism of action of the tyrosine kinase inhibitors gefitinib, lapatinib and neratinib in the treatment of HER2-positive breast cancer: preclinical and clinical evidence.

An increasing number of tumors, including breast cancer, overexpress proteins of the epidermal growth factor receptor (EGFR) family. The interaction between family members activates signaling pathways that promote tumor progression and resistance to treatment. Human epidermal growth factor receptor type II (HER2) positive breast cancer represents a clinical challenge for current therapy. It has motivated the development of novel and more effective therapeutic EGFR family target drugs, such as tyrosine kinase inhibitors (TKIs). This review focuses on the effects of three TKIs mostly studied in HER2- positive breast cancer, lapatinib, gefitinib and neratinib. Herein, we discuss the mechanism of action, therapeutic advantages and clinical applications of these TKIs. To date, TKIs seem to be promising therapeutic agents for the treatment of HER2-overexpressing breast tumors, either as monotherapy or combined with other pharmacological agents.

Control de la variabilidad en la programación de pacientes electivos en salas de cirugía / Variability Control in the Scheduling of Elective Patients in Operating Rooms / Controle da variabilidade no agendamento de pacientes eletivos em salas de cirurgia

El Departamento de Cirugía genera una buena parte de los ingresos de un hospital. Este estudio evalúa la variabilidad en la programación de las cirugías y cómo esta atenta contra la eficiencia del servicio. Este estudio implemento un algoritmo para la generación de dos programaciones que luego fueron comparadas con la realizada manualmente en un hospital en Colombia. Las dos programaciones generadas se diferencian entre sí al incluir o no las máximas desviaciones en los tiempos de duración de las cirugías, asumiendo que estas son causadas por factores externas al proceso y pueden ser eliminadas. Los resultados muestran que al emplear un algoritmo automático para programar las cirugías, podrían mejorarse el tiempo de respuesta y la utilización de las salas. Aún más, se pudo concluir que la variabilidad asociada a la programación manual tiene un mayor impacto que la asociada a otros factores en los indicadores de desempeño del servicio.

The Surgery Department generates a sizeable amount of the income of a hospital. This study evaluates the variability in the scheduling of surgeries and how this threatens the efficiency of the service. This study uses an algorithm for the generation of two schedules that were later compared with a manually-generated schedule in a hospital in Colombia. The difference between the generated schedules is the inclusion or lack thereof of the maximum time deviations in surgery times, under the assumption that these are caused by external factors and may be eliminated. Results show that by using an automatic algorithm to schedule the surgeries response time and operating room use can be improved. Moreover, it can be concluded that the variability associated with manual scheduling has a greater impact than the one associated to other factors in the service performance indicators.

O Departamento de Cirurgia gera uma boa parte das rendas de um hospital. Este estudo avalia a variabilidade no agendamento das cirurgias e como ele atenta contra a eficiencia do servico. Este estudo implementou um algoritmo para a geração de dois agendamentos após comparados com aqueles realizados manualmente em um hospital na Colombia. Os dois agendamentos gerados diferenciam-se entre se por incluir ou nao os máximos afastamentos nos tempos de duração da cirurgia, assumindo que estes sao causados por fatores externos ao processo e podem ser eliminados. Os resultados mostram que usando um algoritmo automático para programar cirurgias, poderia se melhorar o tempo de resposta e a utilização das salas. Além disso, pode-se concluir que a variabilidade associada ao agendamento manual tem maior impacto que a associada a outros fatores nos indicadores de desempenho do servico.

IL-10 inhibits while calcitriol reestablishes placental antimicrobial peptides gene expression.

IL-10 and calcitriol help to achieve a successful pregnancy by suppressing active maternal immunity; however, these factors exert opposite effects upon microbial infections. In the skin and immune cells, IL-10 downregulates ß-defensins while calcitriol induces cathelicidin gene expression in various tissues including placenta. Though, the regulation of human placental ß-defensins by IL-10 and calcitriol has not been studied. Therefore, we explored the regulation of these antimicrobial peptides expression in cultured placental cells by calcitriol and IL-10 alone and combined. Real time PCR showed that calcitriol stimulated, while IL-10 inhibited, ß-defensins and cathelicidin gene expression (P<0.05). In coincubations studies, calcitriol was able to maintain antimicrobial peptides gene expression above control values, overriding IL-10 inhibitory effects. Calcitriol downregulated endogenous IL-10 secretion. Interestingly, calcitriol and TNF-α cooperatively enhanced ß-defensins, while TNF-α reduced basal and calcitriol-stimulated cathelicidin gene expression. In summary, calcitriol and IL-10 exerted opposite effects on antimicrobial peptides expression in the human placenta, suggesting that unbalanced production of IL-10 and calcitriol could be deleterious to innate immune responses during gestation. Our results suggest that calcitriol enhancement of placental defenses involves two mechanisms: (1) downregulation of IL-10 secretion and (2) direct upregulation of ß-defensins and cathelicidin gene expression. Considering that IL-10 and calcitriol differentially regulate the innate immune response in the placenta, in the case of an infection, calcitriol might restrict IL-10 permissive actions towards microbial invasion while restrains inflammation, allowing for pregnancy to continue in quiescence. These results strongly advice maternal vitamin D sufficiency during pregnancy.

In vivo dual targeting of the oncogenic Ether-à-go-go-1 potassium channel by calcitriol and astemizole results in enhanced antineoplastic effects in breast tumors.

BACKGROUND: The oncogenic ether-à-go-go-1 potassium channel (EAG1) activity and expression are necessary for cell cycle progression and tumorigenesis. The active vitamin D metabolite, calcitriol, and astemizole, a promising antineoplastic drug, target EAG1 by inhibiting its expression and blocking ion currents, respectively. We have previously shown a synergistic antiproliferative effect of calcitriol and astemizole in breast cancer cells in vitro, but the effect of this dual therapy in vivo has not been studied. METHODS: In the present study, we explored the combined antineoplastic effect of both drugs in vivo using mice xenografted with the human breast cancer cell line T-47D and a primary breast cancer-derived cell culture (MBCDF). Tumor-bearing athymic female mice were treated with oral astemizole (50 mg/kg/day) and/or intraperitoneal injections of calcitriol (0.03 µg/g body weight twice a week) during 3 weeks. Tumor sizes were measured thrice weekly. For mechanistic insights, we studied EAG1 expression by qPCR and Western blot. The expression of Ki-67 and the relative tumor volume were used as indicators of therapeutic efficacy. RESULTS: Compared to untreated controls, astemizole and calcitriol significantly reduced, while the coadministration of both drugs further suppressed, tumor growth (P < 0.05). In addition, the combined therapy significantly downregulated tumoral EAG1 and Ki-67 expression. CONCLUSIONS: The concomitant administration of calcitriol and astemizole inhibited tumor growth more efficiently than each drug alone, which may be explained by the blocking of EAG1. These results provide the bases for further studies aimed at testing EAG1-dual targeting in breast cancer tumors expressing both EAG1 and the vitamin D receptor.

Calcitriol restores antiestrogen responsiveness in estrogen receptor negative breast cancer cells: a potential new therapeutic approach.

BACKGROUND: Approximately 30% of breast tumors do not express the estrogen receptor (ER) α, which is necessary for endocrine therapy approaches. Studies are ongoing in order to restore ERα expression in ERα-negative breast cancer. The aim of the present study was to determine if calcitriol induces ERα expression in ER-negative breast cancer cells, thus restoring antiestrogen responses. METHODS: Cultured cells derived from ERα-negative breast tumors and an ERα-negative breast cancer cell line (SUM-229PE) were treated with calcitriol and ERα expression was assessed by real time PCR and western blots. The ERα functionality was evaluated by prolactin gene expression analysis. In addition, the effects of antiestrogens were assessed by growth assay using the XTT method. Gene expression of cyclin D1 (CCND1), and Ether-à-go-go 1 (EAG1) was also evaluated in cells treated with calcitriol alone or in combination with estradiol or ICI-182,780. Statistical analyses were determined by one-way ANOVA. RESULTS: Calcitriol was able to induce the expression of a functional ERα in ER-negative breast cancer cells. This effect was mediated through the vitamin D receptor (VDR), since it was abrogated by a VDR antagonist. Interestingly, the calcitriol-induced ERα restored the response to antiestrogens by inhibiting cell proliferation. In addition, calcitriol-treated cells in the presence of ICI-182,780 resulted in a significant reduction of two important cell proliferation regulators CCND1 and EAG1. CONCLUSIONS: Calcitriol induced the expression of ERα and restored the response to antiestrogens in ERα-negative breast cancer cells. The combined treatment with calcitriol and antiestrogens could represent a new therapeutic strategy in ERα-negative breast cancer patients.