A comparison of B16 melanoma cells and 3T3 fibroblasts concerning cell viability and ROS production in the presence of melatonin, tested over a wide range of concentrations

Melatonin is a pleiotropic molecule with many cellular and systemic actions, including chronobiotic effects. Beneficial effects are widely documented concerning the treatment of neoplastic diseases in vivo as well as reductions in viability of cultured cells from melanoma, one of the most aggressive cancers in humans. However, studies of its effects on non-tumor cells in vitro have not focused on viability, except for experiments aiming to protect against oxidotoxicity or other toxicological insults. Furthermore, there is no agreement on the range of effective melatonin concentrations in vitro, and the mechanisms that reduce cell viability have remained unclear. Tumor cell-specific increases in the production of reactive oxygen and nitrogen species (ROS/RNS) may provide a possible explanation. Our aim was to analyze the potential inhibition of tumor (B16 melanoma 4A5) and non-tumor cell (3T3 Swiss albino) viability using a wide range of melatonin concentrations (10-11-10-2 M), and to determine whether intracellular ROS enhancement was involved in this process. In the absence of fetal bovine serum (FBS), low melatonin concentrations (10-9-10-5 M) reduced the proliferation of melanoma cells with no effect in fibroblasts, whereas, in the presence of FBS, they had no effect or even increased the proliferation of both fibroblast and melanoma cells. Melatonin concentrations in the upper millimolar range increased ROS levels and reduced the viability of both cell types, but more markedly so in non-tumor cells. Thus, low melatonin concentrations reduce proliferation in this specific melanoma cell line, whereas high concentrations affect the viability of both tumor (B16 4A5 melanoma) and non-tumor (3T3 fibroblasts) cells. Increased ROS levels in both lines indicate a role for ROS production in the reduction of cell viability at high-but not low-melatonin concentrations, although the mechanism of action still remains to be elucidated. © 2013 by the authors; licensee MDPI, Basel, Switzerland.This project was funded by the Instituto de Salud Carlos III (RETICEF, RD06/0013/0019; RD06/0013/0001 and RD12/0043/0011), the Ministry of Education and Science (BFU2010-21945-C02-01) and a Research fellowship granted to MA Bonmatí (AP2009-1051).Peer Reviewe

Living at the Wrong Time: Effects of Unmatching Official Time in Portugal and Western Spain

Human circadian rhythmicity is subjected to the internal circadian clock, the sun and social clocks (official time, social/work schedules). The discrepancy among these clocks, as occurs when official time does not match its geographical time zone, may produce circadian disruption. Western Spain (GMT+1/+2) and Portugal (GMT0/+1) share similar longitudes (sun time) but have different official times. This provides a unique opportunity to evaluate the effects of official time on circadian rhythmicity and sleep in elderly and retired populations (with no remunerated duties presumed, although other social commitments may be present) at both locations. Although both populations slept enough for their age (7-8 h), circadian robustness (e.g., interdaily stability, relative amplitude) was greater in Portugal, especially during weekdays, while greater desynchronization (both body temperature vs. motor activity and body temperature vs. light exposure) tended to occur in the Spaniards. Once corrected by GMT0, meals took place later in Spain than in Portugal, especially as the day progresses, and a possible interplay between bed/meal timings and internal desynchronization was found. Our results point to the possible deleterious effect on circadian system robustness when official time is misaligned with its geographical time zone.This research was funded by the Ministry of Economy and Competitiveness, the Instituto de Salud Carlos III through a CIBERFES grant (CB16/10/00239, CB16/10/00468); Fundación General del Consejo Superior de Investigaciones Científicas through grant ModulEn (POCTEP 0348_CIE_6_E, Programa de cooperación INTERREG V-A España-Portugal) and Diabfrail LatAm (European Union Horizon 2020 research and innovation programme No. 825546) awarded to MAR (all co-financed by FEDER). Grant RTI2018-093528-B-I00, funded by MCIN/AEI/ 10.13039/501100011033 and by “ERDF A way of making Europe”, by the “European Union” or by the “European Union NextGenerationEU/PRTR”. A research fellowship was granted to MAB-C (20401/SF/17, Fundación Séneca, Región de Murcia (Spain)).S

Chronodisruption and Ambulatory Circadian Monitoring in Cancer Patients: Beyond the Body Clock

Purpose of Review: Circadian rhythms impose daily rhythms a remarkable variety of metabolic and physiological functions, such as cell proliferation, inflammation, and DNA damage response. Accumulating epidemiological and genetic evidence indicates that circadian rhythms’ disruption may be linked to cancer. The integration of circadian biology into cancer research may offer new options for increasing cancer treatment effectiveness and would encompass the prevention, diagnosis, and treatment of this disease. Recent Findings: In recent years, there has been a significant development and use of multi-modal sensors to monitor physical activity, sleep, and circadian rhythms, allowing, for the very first time, scaling accurate sleep monitoring to epidemiological research linking sleep patterns to disease, and wellness applications providing new potential applications. Summary: This review highlights the role of circadian clock in tumorigenesis, cancer hallmarks and introduces the state-of-the-art in sleep-monitoring technologies, discussing the eventual application of insights in clinical settings and cancer research.publishersversionpublishe

Teaching biological rhythms in endocrinology: cortisol and wrist temperature. Póster

This work has been developed as part of the Endocrinology and Metabolism student’s laboratory formation from Biology degree at the University of Murcia. Endocrinology and Metabolism is an elective oneterm course that is taught in the fourth year (4.5 ECTS) for "Biosanitary and Biotechnology" intensification. The course focuses on the study of global and intermediary human metabolism and hormonal regulation, both under normal and special situations. It is our objective to get undergraduate students of Endocrinology and Metabolism with the importance of hormonal diurnal fluctuations in endocrine systems through their involvement in an innovative research program. In addition, with the participation of PhD student in this program, we try to improve their skills in innovative teaching. In humans cortisol circadian rhythm peaks in the morning and shows the lowest levels during the midnight. This fluctuation of cortisol plasma level is reflected in saliva, allowing a simple, non invasive and unstressful sample collection. The influence of different factors, exercise, schedule and weekend shifts, on the rhythmic pattern of cortisol has been studied along various years.Campus Mare Nostrum, Universidad Politécnica de Cartagena, Universidad de Murcia, Región de Murci

Chronodisruption and Ambulatory Circadian Monitoring in Cancer Patients: Beyond the Body Clock

Purpose of Review: Circadian rhythms impose daily rhythms a remarkable variety of metabolic and physiological functions, such as cell proliferation, inflammation, and DNA damage response. Accumulating epidemiological and genetic evidence indicates that circadian rhythms’ disruption may be linked to cancer. The integration of circadian biology into cancer research may offer new options for increasing cancer treatment effectiveness and would encompass the prevention, diagnosis, and treatment of this disease. Recent Findings: In recent years, there has been a significant development and use of multi-modal sensors to monitor physical activity, sleep, and circadian rhythms, allowing, for the very first time, scaling accurate sleep monitoring to epidemiological research linking sleep patterns to disease, and wellness applications providing new potential applications. Summary: This review highlights the role of circadian clock in tumorigenesis, cancer hallmarks and introduces the state-of-the-art in sleep-monitoring technologies, discussing the eventual application of insights in clinical settings and cancer researchThis work was supported in part by CLARIFY project, within European Union’s Horizon 2020 Research and Innovation Programme under grant agreement No. 875160, Instituto de Fomento de la Región de Murcia (INFO) and the European Regional Development Fund (FEDER

Chronodisruption and Ambulatory Circadian Monitoring in Cancer Patients: Beyond the Body Clock.

Purpose of Review Circadian rhythms impose daily rhythms a remarkable variety of metabolic and physiological functions, such as cell proliferation, infammation, and DNA damage response. Accumulating epidemiological and genetic evidence indicates that circadian rhythms’ disruption may be linked to cancer. The integration of circadian biology into cancer research may ofer new options for increasing cancer treatment efectiveness and would encompass the prevention, diagnosis, and treatment of this disease. Recent Findings In recent years, there has been a signifcant development and use of multi-modal sensors to monitor physical activity, sleep, and circadian rhythms, allowing, for the very frst time, scaling accurate sleep monitoring to epidemiological research linking sleep patterns to disease, and wellness applications providing new potential applications. Summary This review highlights the role of circadian clock in tumorigenesis, cancer hallmarks and introduces the stateof-the-art in sleep-monitoring technologies, discussing the eventual application of insights in clinical settings and cancer research.post-print1077 K

Age and Chronodisruption in Mouse Heart: Effect of the NLRP3 Inflammasome and Melatonin Therapy

Age and age-dependent inflammation are two main risk factors for cardiovascular diseases. Aging can also affect clock gene-related impairments such as chronodisruption and has been linked to a decline in melatonin synthesis and aggravation of the NF- B/NLRP3 innate immune response known as inflammaging. The molecular drivers of these mechanisms remain unknown. This study investigated the impact of aging and NLRP3 expression on the cardiac circadian system, and the actions of melatonin as a potential therapy to restore daily rhythms by mitigating inflammaging. We analyzed the circadian expression and rhythmicity of clock genes in heart tissue of wild-type and NLRP3-knockout mice at 3, 12, and 24 months of age, with and without melatonin treatment. Our results support that aging, NLRP3 inflammasome, and melatonin affected the cardiac clock genes expression, except for Rev-erba, which was not influenced by genotype. Aging caused small phase changes in Clock, loss of rhythmicity in Per2 and Rora, and mesor dampening of Clock, Bmal1, and Per2. NLRP3 inflammasome influenced the acrophase of Clock, Per2, and Rora. Melatonin restored the acrophase and the rhythm of clock genes affected by age or NLRP3 activation. The administration of melatonin re-established murine cardiac homeostasis by reversing age-associated chronodisruption. Altogether, these results highlight new findings about the effects aging and NLRP3 inflammasome have on clock genes in cardiac tissue, pointing to continuous melatonin as a promising therapy to placate inflammaging and restore circadian rhythm in heart muscle. Additionally, light microscopy analysis showed age-related morphological impairments in cardiomyocytes, which were less severe in mice lacking NLRP3. Melatonin supplementation preserved the structure of cardiac muscle fibers in all experimental groups.Instituto de Salud Carlos III (Ministerio de Economia y Competitividad, Spain) (European Regional Development Fund/European Social Fund "Investing in your future") PI13-981 PI16-00519 PI19-01372 CB16-10-00238 CB16/10/00239Junta de Andalucia CTS-101Spanish Governmen

Pileflebitis secundaria a brote de enfermedad inflamatoria intestinal

Pylephlebitis or pylethrombophlebitis is a clinical condition defined as the presence of septic thrombophlebitis in the duct-portal vein or intrahepatic vessels which may extend through nearby areas. Etiological diagnosis is crucial since it may modify its therapeutic attitude and is related to intra-abdominal infections. We present the case of a patient with ulcerative colitis who was admitted to the hospital due to an inflammatory bowel disease flare. An abdominal CT-scan showed thrombosis of the splenic-portal system and mesenteric veins which confirmed the diagnosis of pylephlebitis. The patient developed a series of infectious and internal medium complications which led to his death.La pileflebitis o piletromboflebitis es una afección definida como la presencia de tromboflebitis séptica en el tronco venoso portal o ramas intrahepáticas que puede extenderse a territorios cercanos. Su diagnóstico etiológico es esencial ya que puede modificar la actitud terapéutica y su aparición se relaciona con procesos infecciosos intraabdominales. Se presenta el caso de un paciente con colitis ulcerosa que ingresó por brote de enfermedad intestinal. La TC abdominal mostró trombosis del eje esplenoportal y venas mesentéricas que confirmaron el diagnóstico de pileflebitis asociada, presentando el paciente complicaciones infecciosas y de medio interno que desembocaron en su fallecimiento

A New Integrated Variable Based on Thermometry, Actimetry and Body Position (TAP) to Evaluate Circadian System Status in Humans

The disruption of the circadian system in humans has been associated with the development of chronic illnesses and the worsening of pre-existing pathologies. Therefore, the assessment of human circadian system function under free living conditions using non-invasive techniques needs further research. Traditionally, overt rhythms such as activity and body temperature have been analyzed separately; however, a comprehensive index could reduce individual recording artifacts. Thus, a new variable (TAP), based on the integrated analysis of three simultaneous recordings: skin wrist temperature (T), motor activity (A) and body position (P) has been developed. Furthermore, we also tested the reliability of a single numerical index, the Circadian Function Index (CFI), to determine the circadian robustness. An actimeter and a temperature sensor were placed on the arm and wrist of the non-dominant hand, respectively, of 49 healthy young volunteers for a period of one week. T, A and P values were normalized for each subject. A non-parametric analysis was applied to both TAP and the separate variables to calculate their interdaily stability, intradaily variability and relative amplitude, and these values were then used for the CFI calculation. Modeling analyses were performed in order to determine TAP and CFI reliability. Each variable (T, A, P or TAP) was independently correlated with rest-activity logs kept by the volunteers. The highest correlation (r = −0.993, p<0.0001), along with highest specificity (0.870), sensitivity (0.740) and accuracy (0.904), were obtained when rest-activity records were compared to TAP. Furthermore, the CFI proved to be very sensitive to changes in circadian robustness. Our results demonstrate that the integrated TAP variable and the CFI calculation are powerful methods to assess circadian system status, improving sensitivity, specificity and accuracy in differentiating activity from rest over the analysis of wrist temperature, body position or activity alone

A Comparison of B16 Melanoma Cells and 3T3 Fibroblasts Concerning Cell Viability and ROS Production in the Presence of Melatonin Tested in a Wide Concentration Range

Melatonin is a pleiotropic molecule with many cellular and systemic actions, including chronobiotic effects. Beneficial effects are widely documented concerning the treatment of neoplastic diseases in vivo as well as reductions in viability of cultured cells from melanoma, one of the most aggressive cancers in humans. However, studies of its effects on non-tumor cells in vitro have not focused on viability, except for experiments aiming to protect against oxidotoxicity or other toxicological insults. Furthermore, there is no agreement on the range of effective melatonin concentrations in vitro, and the mechanisms that reduce cell viability have remained unclear. Tumor cell-specific increases in the production of reactive oxygen and nitrogen species (ROS/RNS) may provide a possible explanation. Our aim was to analyze the potential inhibition of tumor (B16 melanoma 4A5) and non-tumor cell (3T3 Swiss albino) viability using a wide range of melatonin concentrations (10(−11)–10(−2) M), and to determine whether intracellular ROS enhancement was involved in this process. In the absence of fetal bovine serum (FBS), low melatonin concentrations (10(−9)–10(−5) M) reduced the proliferation of melanoma cells with no effect in fibroblasts, whereas, in the presence of FBS, they had no effect or even increased the proliferation of both fibroblast and melanoma cells. Melatonin concentrations in the upper millimolar range increased ROS levels and reduced the viability of both cell types, but more markedly so in non-tumor cells. Thus, low melatonin concentrations reduce proliferation in this specific melanoma cell line, whereas high concentrations affect the viability of both tumor (B16 4A5 melanoma) and non-tumor (3T3 fibroblasts) cells. Increased ROS levels in both lines indicate a role for ROS production in the reduction of cell viability at high—but not low—melatonin concentrations, although the mechanism of action still remains to be elucidated