In vitro and in vivo effects of lutein against cisplatin-induced ototoxicity

This is peer reviewed version of the following article Experimental and Toxicologic Pathology 68.4 (2016): 197-204, which has been published in final form at http://dx.doi.org/10.1016/j.etp.2016.01.003Introduction: Cisplatin is a commonly prescribed drug that produces ototoxicity as a side effect. Lutein is a carotenoid with antioxidant and anti-inflammatory properties previously tested for eye, heart and skin diseases but not evaluated to date in ear diseases. Aim: To evaluate the protective effects of lutein on HEI-OC1 auditory cell line and in a Wistar rat model of cisplatin ototoxicity. Materials and Methods: In vitro study: Culture HEI-OC1 cells were exposed to lutein (2.5-100 μM) and to 25 μM cisplatin for 24 h. In vivo study: Twenty eight female Wistar rats were randomized into three groups. Group A (n = 8) received intratympanic lutein (0.03 mL) (1 mg/mL) in the right ear and saline solution in the left one to determine the toxicity of lutein. Group B (n = 8) received also intraperitoneal cisplatin (10 mg/kg) to test the efficacy of lutein against cisplatin ototoxicity. Group C (n = 12) received intratympanic lutein (0.03 mL) (1 mg/mL) to quantify lutein in cochlear fluids (30 min, 1 h and 5 days after treatment). Hearing function was evaluated by means of Auditory Steady-State Responses before the procedure and 5 days after (groups A and B). Morphological changes were studied by confocal laser scanning microscopy. Results: In vitro study: Lutein significantly reduced the cisplatin-induced cytotoxicity in the HEI-OC1 cells when they were pre-treated with lutein concentrations of 60 and 80 μM. In vivo study: Intratympanic lutein (1 mg/mL) application showed no ototoxic effects. However it did not achieve protective effect against cisplatin-induced ototoxicity in Wistar rats. Conclusions: Although lutein has shown beneficial effects in other pathologies, the present study only obtained protection against cisplatin ototoxicity in culture cells, but not in the in vivo model. The large molecule size, the low dose administered, and restriction to diffusion in the inner ear could account for this negative result.Research supported by a Spanish FIS Grant EI 11/00742

Clonal chromosomal mosaicism and loss of chromosome Y in elderly men increase vulnerability for SARS-CoV-2

The pandemic caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2, COVID-19) had an estimated overall case fatality ratio of 1.38% (pre-vaccination), being 53% higher in males and increasing exponentially with age. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, we found 133 cases (1.42%) with detectable clonal mosaicism for chromosome alterations (mCA) and 226 males (5.08%) with acquired loss of chromosome Y (LOY). Individuals with clonal mosaic events (mCA and/or LOY) showed a 54% increase in the risk of COVID-19 lethality. LOY is associated with transcriptomic biomarkers of immune dysfunction, pro-coagulation activity and cardiovascular risk. Interferon-induced genes involved in the initial immune response to SARS-CoV-2 are also down-regulated in LOY. Thus, mCA and LOY underlie at least part of the sex-biased severity and mortality of COVID-19 in aging patients. Given its potential therapeutic and prognostic relevance, evaluation of clonal mosaicism should be implemented as biomarker of COVID-19 severity in elderly people. Among 9578 individuals diagnosed with COVID-19 in the SCOURGE study, individuals with clonal mosaic events (clonal mosaicism for chromosome alterations and/or loss of chromosome Y) showed an increased risk of COVID-19 lethality

Time and space resolved optical emission diagnostics of laser induced breakdown muscle tissue samples

XXIV Reunión Nacional de Espectroscopia-VIII Congreso Ibérico de Espectroscopia ; organizan Sociedad de Espectroscopia Aplicada (SEA) y la Universidad de la Rioja. P100The recent progress made in developing laser-induced breakdown spectroscopy (LIBS) has transformed this technique from an elemental analysis method to one that can be applied for the analysis of complex biological material or clinical specimens. The LIBS method has gained a reputation as a flexible and convenient technique for rapidly identification of unknown materials (chemical, biological or explosive). The plasma generated by LIBS technique on muscle tissue samples [1] was investigated using two high-power pulsed lasers (transverse excitation atmospheric CO2 and Nd:YAG lasers). A remarkable fact is the no influence of the laser wavelength on the observed spectral lines and molecular bands. The emission of the plasma shows excited neutral Na, K, C, Mg, H, N and O atoms, ionized C+, C2+, Mg+, N+ and O+ species and molecular band systems of CN(B2¿+ ¿ X2¿+), C2(d3¿g ¿ a3¿u), CH(A2¿ ¿ X2¿), NH(A3¿ ¿ X3¿-) and OH(A2¿ ¿ X2¿). For the assignment of the atomic/ionic lines we used the information tabulated in NIST [2]. The molecular bands were compared with the LIBS experiments obtained in our laboratory on different samples [3-6]. We focus our attention on the dynamics of the muscle tissue laser induced plasma species expanding into air (atmospheric pressure) or into vacuum (air pressures of 0.8 and 0.01 Pa). In conventional one dimensional optical emission spectroscopy (OES) studies, various plasma-plume segments were selected along the plume expansion axis and averaged over line-of-sight. This setup was easily transformed to a two-dimensional (2D) OES setup [7] by inserting a Dove prism between the focusing and collimating lenses. Time-integrated and time-resolved 2D OES plasma profiles were recorded as a function of emitted wavelength and distance from the target. Different plasma parameters such as velocities, temperatures and electron densities were evaluated using OES. The temporal behaviour of specific lines of atomic/ionic lines was characterized.This work was partially supported by the MICINN (Spain, Ministerio de Ciencia e Innovación), project CTQ2010-15680, Autónoma University of Madrid, project CEMU-2012-003 and Complutense University of Madrid, grant CCG10-UCM/PPQ-4713.Peer Reviewe

Relationship between Fertility Traits and Kinematics in Clusters of Boar Ejaculates

The aim was to determine the relationship between kinematic parameters of boar spermatozoa and fertility rates of sow, as well as to assess the effect of sperm clusters on the fertility capacity of the ejaculate. Semen samples were collected from 11 sexually mature boars. Samples were analyzed by an ISAS®v1 CASA-Mot system for eight kinematic parameters. Ejaculate clusters were characterized using multivariate procedures, such as principal factors (PFs) analysis and clustering methods (the k-means model). Four different ejaculate clusters were identified from two kinematic PFs which involved linear trajectory and velocity. There were differences (p 0.05) between dam lines and ejaculate clusters in fertility variables. The discriminant ability of the different kinematics of sperm variables to predict litter size fertility was analyzed using receiver operating characteristics (ROC) curve analysis. Curvilinear velocity (VCL), average path velocity (VAP), amplitude of lateral head displacement (ALH), and beat-cross frequency (BCF) showed significant, albeit limited, predictive capacity for litter size fertility variables (range: 0.55–0.58 area under curve, AUC). The kinematic analysis of the ejaculates in clusters did not have a predictive capacity for litter size variables.This research was funded by Fundación para el Fomento y Promoción de la Investigación y Transferencia de Tecnología Agropecuaria de Costa Rica (FITTACORI), grant number F23-19 and the Costa Rica Institute of Technology (Vice-Chancellor’s office of Research and Extension; VIE (Vicerrectoría de investigación y Extensión; Project-VIE-5402-2151-1015).Peer reviewe

Sperm kinematic subpopulations of the American crocodile (Crocodylus acutus)

There has been very limited use of computer assisted semen analysis (CASA) to evaluate reptile sperm. The aim of this study was to examine sperm kinematic variables in American crocodile (Crocodylus acutus) semen samples and to assess whether sperm subpopulations could be characterized. Eight ejaculates (two ejaculates/male) from four sexually mature captive crocodiles were obtained. An ISAS®v1 CASA-Mot system, with an image acquisition rate of 50 Hz, and ISAS®D4C20 counting chambers were used for sperm analyses. The percentages of motile and progressively motile spermatozoa did not differ among animals (P > 0.05) but there was a significant animal effect with regards to kinematic variables (P < 0.05). Principal component (PC) analysis revealed that kinematic variables grouped into three components: PC1, related to velocity; PC2 to progressiveness and PC3 to oscillation. Subpopulation structure analysis identified four groups (P < 0.05), which represented, on average, 9.8%, 32.1%, 26.8%, and 31.3% of the total sperm population. Males differed in the proportion of sperm in each of the kinematic subpopulations. This new approach for the analysis of reptile sperm kinematic subpopulations, reflecting quantifiable parameters generated by CASA system technology, opens up possibilities for future assessments of crocodile sperm and will be useful in the future development of assisted reproduction for these species.This research was funded by the Costa Rica Institute of Technology (grant number 07-0336-01 to AV), Vice-Chancellor’s office of Research and Extension; VIE, and Agronomy School, and by the Spanish Ministry of Science and Innovation (grant number CGL2016-80577-P to ERSR).We acknowledge support of the publication fee by the CSIC Open Access Publication Support Initiative through its Unit of Information Resources for Research (URICI).Peer reviewe

In vitro and in vivo effects of lutein against cisplatin-induced ototoxicity

[Introduction]: Cisplatin is a commonly prescribed drug that produces ototoxicity as a side effect. Lutein is a carotenoid with antioxidant and anti-inflammatory properties previously tested for eye, heart and skin diseases but not evaluated to date in ear diseases. [Aim]: To evaluate the protective effects of lutein on HEI-OC1 auditory cell line and in a Wistar rat model of cisplatin ototoxicity. [Materials and Methods]: In vitro study: Culture HEI-OC1 cells were exposed to lutein (2.5–100 μM) and to 25 μM cisplatin for 24 h. In vivo study: Twenty eight female Wistar rats were randomized into three groups. Group A (n = 8) received intratympanic lutein (0.03 mL) (1 mg/mL) in the right ear and saline solution in the left one to determine the toxicity of lutein. Group B (n = 8) received also intraperitoneal cisplatin (10 mg/kg) to test the efficacy of lutein against cisplatin ototoxicity. Group C (n = 12) received intratympanic lutein (0.03 mL) (1 mg/mL) to quantify lutein in cochlear fluids (30 min, 1 h and 5 days after treatment). Hearing function was evaluated by means of Auditory Steady-State Responses before the procedure and 5 days after (groups A and B). Morphological changes were studied by confocal laser scanning microscopy. [Results]: In vitro study: Lutein significantly reduced the cisplatin-induced cytotoxicity in the HEI-OC1 cells when they were pre-treated with lutein concentrations of 60 and 80 μM. In vivo study: Intratympanic lutein (1 mg/mL) application showed no ototoxic effects. However it did not achieve protective effect against cisplatin-induced ototoxicity in Wistar rats. [Conclusions]: Although lutein has shown beneficial effects in other pathologies, the present study only obtained protection against cisplatin ototoxicity in culture cells, but not in the in vivo model. The large molecule size, the low dose administered, and restriction to diffusion in the inner ear could account for this negative result.Research supported by a Spanish FIS GrantEI 11/00742.Peer Reviewe