The overlap with metabolic dysfunction-associated steatotic liver disease negatively affects outcomes of primary biliary cholangitis.

BACKGROUND AND AIMS: The relationship between primary biliary cholangitis (PBC) and metabolic dysfunction-associated steatotic liver disease, and its impact on treatment response and prognosis, remains underexplored. METHODS: Patient cohort from two centres comprising long-term follow-up data. All patients had histologically confirmed PBC. Biopsies were classified according to Non-Alcoholic Steatohepatitis Clinical Research Network. Diagnosis of metabolic dysfunction-associated steatotic liver disease was established when steatosis exceeded 5%, along with at least one metabolic risk factor. Patients with specific aetiologies of steatosis, other liver diseases, incomplete results and inadequate treatment with ursodeoxycholic acid were excluded. Data from patients initiating second-line treatment were censored. Treatment response was assessed using the Toronto, Paris II and AST-to-platelet at 12-month criteria. The UK PBC and Globe scores, and liver events were utilized as outcome measures. RESULTS: The study included 129 patients, 36 showing histologically confirmed overlap between PBC and steatosis. Patients with overlap showed worse prognosis according to Paris II (61.1% vs. 33.3%, p = 0.004), Toronto (52.5% vs. 24.7%, p = 0.002), AST-to-platelet 12-month >0.54 (36.1% vs. 17.2%, p = 0.021), Globe >0.30 (49.2% vs. 29.2%, p = 0.033) and UK PBC at 5, 10 and 15 years (p ≤ 0.001). Liver-related mortality and liver transplant were more prevalent in the overlap group (p = 0.001). In the multivariate analysis, steatosis, dyslipidaemia and advanced fibrosis were independently associated to worse outcomes. CONCLUSIONS: Our findings suggest that metabolic dysfunction-associated steatotic liver disease worsens the prognosis of PBC.

Comparative Analysis of Microsatellite and SNP Markers for Genetic Management of Red Deer.

The analysis of population genetic structure and individual multilocus heterozygosity are crucial for wildlife management and conservation. Microsatellite markers have traditionally been used to assess these genetic parameters. However, single-nucleotide polymorphisms (SNPs) are becoming increasingly popular. Our goal here was to determine to what extent SNPs can provide better insights than microsatellites into the overall genetic status and population genetic processes in the species. To this end, we genotyped 210 red deer (Cervus elaphus) in the Spanish wild population with both 11 microsatellites and 31,712 SNPs. We compared parameters related to population genetic structure and individual multilocus heterozygosity obtained with both types of markers. Our results showed correlations between parameters measured using both microsatellites and SNPs, particularly those related to the level of genetic diversity and genetic differentiation. However, we found notably lower precision of microsatellites in measuring the distribution of genetic diversity among individuals. We conclude that microsatellites can be used to monitor the overall genetic status and detect broad patterns in red deer populations. Nevertheless, the greater precision of SNPs in inferring genetic structure and multilocus heterozygosity leads us to encourage scientists and wildlife managers to prioritize their use whenever possible.

Is the Intrasexual Competition in Male Red Deer Reflected in the Ratio of Stable Isotopes of Carbon and Nitrogen in Faeces?

Isotopic analysis of carbon and nitrogen in faeces is a reliable methodology for studying ecology in wildlife. Here, we tested this technique to detect variations in carbon and nitrogen isotopic ratios (δ13C and δ15N) in two different intrasexual competition scenarios of male Iberian red deer (Cervus elaphus hispanicus) using faeces of individuals collected during hunting actions in South-eastern Spain. The carbon isotopic ratio (δ13C) was not found to be significant, likely due to similar diet composition in all individuals. However, the nitrogen isotopic ratio (δ15N) was found to be lower in populations where sexual competition between males during the rut was higher compared to low-competition populations. Therefore, this study suggests a different use of proteins by an individual male red deer depending on the sexually competitive context in which he lives. Although further research is needed, these results show the potential of isotopic analysis as a tool for studying individual and populational variations in the level of intrasexual competition, with implications in evolutionary ecology and population management.

Exploring the potential links between gut microbiota composition and natural populations management in wild boar (Sus scrofa).

We surveyed wild boar (Sus scrofa) populations using 16S rRNA gene analysis of the gut microbiota in fresh faeces taken from 88 animals hunted in 16 hunting estates. The wild boar is a very convenient model system to explore how environmental factors including game management, food availability, disease prevalence, and behaviour may affect different biological components of wild individuals with potential implications in management and conservation. We tested the hypotheses that diet (according to stable carbon isotopes analyses), gender (i.e., animal behaviour studying males and females), and both health (analyses of serum samples to detect exposure to several diseases) and form statutes (i.e., thoracic circumference in adults) are reflected in changes in the intestinal microbiota. We focused on a gut functional biomarker index combining Oscillospiraceae and Ruminococcaceae vs. Enterobacteriaceae. We found that gender and the estate (population) were explanatory variables (c.a. 28% of the variance), albeit a high degree of overlapping among individuals was observed. The individuals with higher abundance of Enterobacteriaceae showed a gut microbiota with low diversity, mostly in males. Significant statistical differences for thoracic circumference were not found between males and females. Interestingly, the thoracic circumference was significantly and inversely related to the relative abundance of Enterobacteriaceae in males. Overall, we found that diet, gender, and form status were major factors that could be related to the composition and diversity of the gut microbiota. A high variability was observed in the biomarker index for populations with natural diet (rich in C3 plants). Although, we noticed a marginally significant negative trend between the index (higher abundance of Enterobacteriaceae) and the continuous feeding of C4 plants (i.e., supplementary maize) in the diet of males. This result suggests that continuous artificial feeding in hunting estates could be one of the factors negatively influencing the gut microbiota and the form status of wild boars that deserves further investigations.

Advances in Transversal Topics Applicable to the Care of Bladder Cancer Patients in the Real-World Setting.

Recommendations regarding transversal topics applicable to bladder cancer patients independent of tumor grade and stage were established by members of the Spanish Oncology Genitourinary Multidisciplinary Working Group (SOGUG). Liquid biopsy in urine and blood samples is useful in the surveillance of non-muscle-invasive and muscle-invasive bladder cancer, respectively. Multiparametric MRI is an accurate, faster and non-invasive staging method overcoming the understaging risk of other procedures. The combination of FDG-PET/MRI could improve diagnostic reliability, but definite criteria for imaging interpretation are still unclear. Hospital oncology pharmacists as members of tumor committees improve the safety of drug use. Additionally, safety recommendations during BCG preparation should be strictly followed. The initial evaluation of patients with bladder cancer should include a multidimensional geriatric assessment. Orthotopic neobladder reconstruction should be offered to motivated patients with full information of self-care requirements. Bladder-sparing protocols, including chemoradiation therapy and immune checkpoints inhibitors (ICIs), should be implemented in centers with well-coordinated multidisciplinary teams and offered to selected patients. The optimal strategy of treatment with ICIs should be defined from the initial diagnostic phase with indications based on scientific evidence. Centralized protocols combined with the experience of professional groups are needed for the integral care of bladder cancer patients.

Stable carbon and nitrogen isotope values in hair reveal management differences and hidden practices in wild boar populations.

The analysis of stable isotopes in different tissues has been widely used to obtain information on the ecology and nutritional patterns of wildlife. The isotope ratios of the stable isotopes of carbon and nitrogen (δ13C and δ15N) analysed in different tissues are directly related to the animal's diet and, to some extent, to the environment where the individual has growth. Specifically, this type of analysis in hair samples has become relevant as it provides information on the quality and long-term composition of the diet that produced the isotope accumulation during the tissue growth. We took samples of wild boar (Sus scrofa) hair from 7 different populations in the south-west of Spain (Mediterranean habitats), in the 2018/2019 hunting season. The main objectives of this study were (i) to investigate the use of hair stable isotopes to reveal differences in composition and quality of the diet of wild boar within the same population or between populations, and (ii) to use hair isotopes as a tool to uncover hidden management practices that may occur in hunting areas associated with the use of supplementary feeding or even captive breeding and release. Each animal had a hair (long 10 cm) analysed in duplicate, previously cut into parts of equal size (from the oldest part of the hair to the most recent part), that were analysed separately. We found differences in δ13C and δ 15N between hair parts and populations, which can be related to management actions at different times during the hair growth. Moreover, the use of corn, a type of plant not occurring naturally in the study area, can be documented with the isotope analysis to prove unauthorized supplementary feeding or captive origin of wild boar in hunting areas.

Intertwined leukocyte balances in tumours and peripheral blood as robust predictors of right and left colorectal cancer survival.

BACKGROUND: Colorectal cancer (CRC) accounts for 9.4% of overall cancer deaths, ranking second after lung cancer. Despite the large number of factors tested to predict their outcome, most patients with similar variables show big differences in survival. Moreover, right-sided CRC (RCRC) and left-sided CRC (LCRC) patients exhibit large differences in outcome after surgical intervention as assessed by preoperative blood leukocyte status. We hypothesised that stronger indexes than circulating (blood) leukocyte ratios to predict RCRC and LCRC patient outcomes will result from combining both circulating and infiltrated (tumour/peritumour fixed tissues) concentrations of leukocytes. AIM: To seek variables involving leukocyte balances in peripheral blood and tumour tissues and to predict the outcome of CRC patients. METHODS: Sixty-five patients diagnosed with colon adenocarcinoma by the Digestive Surgery Service of the La Paz University Hospital (Madrid, Spain) were enrolled in this study: 43 with RCRC and 22 with LCRC. Patients were followed-up from January 2017 to March 2021 to record overall survival (OS) and recurrence-free survival (RFS) after surgical interventions. Leukocyte concentrations in peripheral blood were determined by routine laboratory protocols. Paraffin-fixed samples of tumour and peritumoural tissues were assessed for leukocyte concentrations by immunohistochemical detection of CD4, CD8, and CD14 marker expression. Ratios of leukocyte concentration in blood and tissues were calculated and evaluated for their predictor values for OS and RFS with Spearman correlations and Cox univariate and multivariate proportional hazards regression, followed by the calculation of the receiver-operating characteristic and area under the curve (AUC) and the determination of Youden's optimal cutoff values for those variables that significantly correlated with either RCRC or LCRC patient outcomes. RCRC patients from the cohort were randomly assigned to modelling and validation sets, and clinician-friendly nomograms were developed to predict OS and RFS from the respective significant indexes. The accuracy of the model was evaluated using calibration and validation plots. RESULTS: The relationship of leukocyte ratios in blood and peritumour resulted in six robust predictors of worse OS in RCRC: CD8+ lymphocyte content in peritumour (CD8pt, AUC = 0.585, cutoff < 8.250, P = 0.0077); total lymphocyte content in peritumour (CD4CD8pt, AUC = 0.550, cutoff < 10.160, P = 0.0188); lymphocyte-to-monocyte ratio in peritumour (LMRpt, AUC = 0.807, cutoff < 3.185, P = 0.0028); CD8+ LMR in peritumour (CD8MRpt, AUC = 0.757, cutoff < 1.650, P = 0.0007); the ratio of blood LMR to LMR in peritumour (LMRb/LMRpt, AUC = 0.672, cutoff > 0.985, P = 0.0244); and the ratio of blood LMR to CD8+ LMR in peritumour (LMRb/CD8MRpt, AUC = 0.601, cutoff > 1.485, P = 0.0101). In addition, three robust predictors of worse RFS in RCRC were found: LMRpt (AUC = 0.737, cutoff < 3.185, P = 0.0046); LMRb/LMRpt (AUC = 0.678, cutoff > 0.985, P = 0.0155) and LMRb/CD8MRpt (AUC = 0.615, cutoff > 1.485, P = 0.0141). Furthermore, the ratio of blood LMR to CD4+ LMR in peritumour (LMRb/CD4MRpt, AUC = 0.786, cutoff > 10.570, P = 0.0416) was found to robustly predict poorer OS in LCRC patients. The nomograms showed moderate accuracy in predicting OS and RFS in RCRC patients, with concordance index of 0.600 and 0.605, respectively. CONCLUSION: Easily obtainable variables at preoperative consultation, defining the status of leukocyte balances between peripheral blood and peritumoural tissues, are robust predictors for OS and RFS of both RCRC and LCRC patients.

Computational FEM Model and Phantom Validation of Microwave Ablation for Segmental Microcalcifications in Breasts Using a Coaxial Double-Slot Antenna.

INTRODUCTION: Cancer is the second leading cause of death worldwide. Breast cancer is the second most common cause of cancer-related mortality, accounting for 11.6% of the total number of deaths. The main treatments for this disease are surgical removal of the tumor, radiotherapy, and chemotherapy. Recently, different minimally invasive technologies have been applied (e.g., emission of electromagnetic waves, thermal and chemical means) to overcome the important side effects of these treatment modalities. The objective of this study was to develop and evaluate a predictive computational model of microwave ablation. MATERIALS AND METHODS: The predictive computational model of microwave ablation was constructed by means of a dual-slot coaxial antenna. The model was compared with an experiment performed using a breast phantom, which emulates the dielectric properties of breast tissue with segmental microcalcifications. The standing wave ratio (SWR) was obtained for both methods to make a comparison and determine the feasibility of applying electromagnetic ablation to premalignant lesions in breasts. Specifically, for the analysis of segmental microcalcifications, a breast phantom with segmental microcalcifications was developed and two computational models were performed under the same conditions (except for blood perfusion, which was excluded in one of the models). RESULTS: The SWR was obtained by triplicate experiments in the phantom, and the measurements had a difference of 0.191 between the minimum and maximum SWR values, implying a change of power reflection of 0.8%. The average of the three measurements was compared with the simulation that did not consider blood perfusion. The comparison yielded a change of 0.104, representing a 0.2% change in power reflection. Discussion. Both experimentation in phantom and simulations demonstrated that ablation therapy can be performed using this antenna. However, an additional optimization procedure is warranted to increase the efficiency of the antenna.

Rainfall decrease and red deer rutting behaviour: Weaker and delayed rutting activity though higher opportunity for sexual selection.

In the last decades, climate change has caused an increase in mean temperatures and a reduction in average rainfall in southern Europe, which is expected to reduce resource availability for herbivores. Resource availability can influence animals' physical condition and population growth. However, much less is known on its effects on reproductive performance and sexual selection. In this study, we assessed the impact of three environmental factors related to climate change (rainfall, temperature and vegetation index) on Iberian red deer Cervus elaphus hispanicus reproductive timing and sexual behaviour, and their effects on the opportunity for sexual selection in the population. We measured rutting phenology as rut peak date, the intensity of male rutting activity as roaring rate, and the opportunity for sexual selection from the distribution of females among harem holding males in Doñana Biological Reserve (Southwest Spain), from data of daily observations collected during the rut over a period of 25 years. For this study period, we found a trend for less raining and hence poorer environmental conditions, which associated with delayed rutting season and decreased rutting intensity, but that appeared to favour a higher degree of polygyny and opportunity for sexual selection, all these relationships being modulated by population density and sex ratio. This study highlights how climate change (mainly rainfall reduction in this area) can alter the conditions for mating and the opportunity for sexual selection in a large terrestrial mammal.

The dark-ventral-patch of male red deer, a sexual signal that conveys the degree of involvement in rutting behavior.

BACKGROUND: In polygynous mammals, signalling may play a decisive role in mating behavior, mediating the intensity of male fights and female mate choice. During the rutting season, male red deer may show a visible dark patch in their ventral fur. Recently, this patch has been suggested to act as a flexible sexual signal, due to its relationships with other variables such as age, body size, antler development, volatile compounds, or the competitive environment. The analysis of fur pigmentation at the ventral patch suggests that this might also visually indicate the male intrinsic predisposition to take part in mating competition. RESULTS: To assess the possible role of this trait as a communicative signal related to mate competition, we used red deer behavioral observations during the rut in Doñana National Park (Spain) to examine the link between the degree of expression of the dark ventral patch and the rutting activity (assessed from both intra-and-inter-sexual behaviors). Consistent with our predictions, we found in a field study that males with large dark patches showed a higher frequency of rutting behaviors (mainly roaring and flehmen), more interactions with females, and attained larger harem sizes. CONCLUSIONS: The dark ventral patch was a better predictor of male behavior than antler tines or territory holding, thus standing as a short-term indicator of male willingness to invest in mating competition.

Contrasting Environmental Drivers Determine Biodiversity Patterns in Epiphytic Lichen Communities along a European Gradient.

Assessing the ecological impacts of environmental change on biological communities requires knowledge of the factors driving the spatial patterns of the three diversity facets along extensive environmental gradients. We quantified the taxonomic (TD), functional (FD), and phylogenetic diversity (PD) of lichen epiphytic communities in 23 beech forests along Europe to examine their response to environmental variation (climate, habitat quality, spatial predictors) at a continental geographic scale. We selected six traits related to the climatic conditions in forest ecosystems, the water-use strategy and the nutrient uptake, and we built a phylogenetic tree based on four molecular markers. FD and climate determined TD and PD, with spatial variables also affecting PD. The three diversity facets were primarily shaped by distinct critical predictors, with the temperature diurnal range affecting FD and PD, and precipitation of the wettest month determining TD. Our results emphasize the value of FD for explaining part of TD and PD variation in lichen communities at a broad geographic scale, while highlighting that these diversity facets provide complementary information about the communities' response under changing environmental conditions. Furthermore, traits such as growth form, photobiont type, and reproductive strategy mediated the response of lichen communities to abiotic factors emerging as useful indicators of macroclimatic variations.

The dark ventral patch: A bimodal flexible trait related to male competition in red deer.

Sexual signals play a central role in male-male competition in polygynous species. In red deer (Cervus elaphus), male's ventral area become dark during the rutting season due to urine spraying behaviour and retains many chemical compounds potentially revealing individual features. Here we investigate the variation in size of this trait, exploring its relationship with age and male competitive features such as antlers or body size, as well as populational level of intrasexual competition for mates. We found that the size of the dark ventral patch followed a clearly bimodal distribution, i.e. males mostly expressed the full-size trait or just developed a very small one. For these two groups of males according to trait expression, the relationships of trait size with age and antler size differed. Populational level of intrasexual competition appeared to affect the relationship between antler size and the probability of a fully developed ventral patch. These results indicate that the trait encodes information on body size, antler size, age and populational level of mate competition, thus suggesting a role in signalling male's competitive features and willingness to allocate reproductive effort within a particular season.

Estudio observacional de la toxicidad con diferentes formulaciones de docetaxel en pacientes con cáncer de mama / Observational study of clinical toxicity with different formulations of docetaxel in breast cancer patients

OBJETIVO: Estudiar los excipientes e impurezas de los diferentes medicamentos comercializados de docetaxel y conocer la incidencia de los diversos eventos adversos derivados del uso de docetaxel y su repercusión clínica en pacientes con cáncer de mama en el contexto de adyuvancia o neoadyuvancia. MÉTODO: Estudio observacional, longitudinal, prospectivo y multicéntrico en 26 hospitales de Madrid, Cataluña, Andalucía y Comunidad Valenciana. Se caracterizaron las distintas formulaciones de docetaxel en cuanto a pH, cantidad de docetaxel e impurezas. Se evaluó la incidencia acumulada de eventos adversos de cualquier grado estratificados por tipo de medicamento, analizando las diferencias mediante el test de χ2.RESULTADOS: Se detectaron diferencias estadísticamente significativas entre las distintas formulaciones de docetaxel en cuanto a la incidencia acumulada por ciclo de: modificación de dosis, anemia, reacciones de hipersensibilidad y anafilaxia, neuropatía, toxicidad palmo-plantar y dermatológica, toxicidad ungueal y edema facial. La formulación con un menor contenido en impurezas presentó mejores resultados en modificación de dosis, visitas a urgencias, e incidencia de anemia y edema facial, pero peores en hospitalización, neutropenia febril, neuropatía motora y toxicidad palmo-plantar. CONCLUSIONES: Los resultados muestran diferencias en la incidencia de los eventos adversos de los distintos medicamentos con docetaxel comercializados en nuestro país, con diferencias significativas entre ellos en algunas de las variables estudiadas. No se ha podido identificar un medicamento con un mejor perfil de toxicidad. Tampoco se ha podido establecer su relación con respecto a la composición de excipientes e impurezas

OBJECTIVE: To analyze the excipients and impurities contained in the various docetaxel products available on the market and find out whether they may be responsible for any of the different adverse events associated with the use of docetaxel in patients with breast cancer receiving adjuvant or neoadjuvant treatment. METHOD: This is a prospective, multicenter, longitudinal observational, study carried in 26 hospitals in Madrid, Catalonia, Andalusia, and the Valencia Region. The different docetaxel formulations were characterized in terms of their pH, amount of the active ingredient and impurities. The cumulative incidence of adverse events of any grade was evaluated. Adverse events were stratified by drug type and differences were analyzed by means of a chi-square test. RESULTS: Statistically significant differences were found between the different docetaxel formulations in the cumulative per-cycle incidence of: dosage change, anemia, hypersensitivity reactions and anaphylaxis, neuropathy, palmoplantar and dermal toxicity, ungual toxicity and facia edema. The formulation with the lowest content of impurities showed better results in terms of change of dosage, visits to the emergency room and incidence of anemia and facial edema. However, it was associated with poorer results regarding hospitalization, febrile neutropenia, motor neuropathy and palmoplantar toxicity. CONCLUSIONS: The results of the study showed differences in the incidence of adverse events of the different docetaxel products available in Spain. Such differences were statistically significant for some of the variables analyzed. The study was not able to determine which of the products offered the best toxicity profile. Nor was it possible to establish a correlation with respect to the composition of excipients or the content of impurities

Observational study of clinical toxicity with different formulations of docetaxel in breast cancer patients.

OBJECTIVE: To analyze the excipients and impurities contained in the various docetaxel products available on the market and find out  whether they may be responsible for any of the different adverse events  associated with the use of docetaxel in patients with breast cancer  receiving adjuvant or neoadjuvant treatment. METHOD: This is a prospective, multicenter, longitudinal observational, study carried in 26 hospitals in Madrid, Catalonia, Andalusia, and the Valencia Region. The different docetaxel formulations were  characterized in terms of their pH, amount of the active ingredient and  impurities. The cumulative incidence of adverse events of any grade was  evaluated. Adverse events were stratified by drug type and differences  were analyzed by means of a chi-square test. RESULTS: Statistically significant differences were found between the different docetaxel formulations in the cumulative per-cycle incidence of: dosage change, anemia, hypersensitivity reactions and  anaphylaxis, neuropathy, palmoplantar and dermal toxicity, ungual toxicity  and facial edema. The formulation with the lowest content of impurities  showed better results in terms of change of dosage, visits to the  emergency room and incidence of anemia and facial edema. However, it  was associated with poorer results regarding hospitalization, febrile  neutropenia, motor neuropathy and palmoplantar toxicity. CONCLUSIONS: The results of the study showed differences in the incidence of adverse events of the different docetaxel products available in  Spain. Such differences were statistically significant for some of the  variables analyzed. The study was not able to determine which of the  products offered the best toxicity profile. Nor was it possible to establish a  correlation with respect to the composition of excipients or the content of  impurities.

Objetivo: Estudiar los excipientes e impurezas de los diferentes  medicamentos comercializados de docetaxel y conocer la incidencia de  los diversos eventos adversos derivados del uso de docetaxel y su  repercusión clínica en pacientes con cáncer de mama en el contexto de  adyuvancia o neoadyuvancia.Método: Estudio observacional, longitudinal, prospectivo y multicéntrico en 26 hospitales de Madrid, Cataluña, Andalucía y  Comunidad Valenciana. Se caracterizaron las distintas formulaciones de  docetaxel en cuanto a pH, cantidad de docetaxel e impurezas. Se evaluó  la incidencia acumulada de eventos adversos de cualquier grado  estratificados por tipo de medicamento, analizando las diferencias  mediante el test de χ2.Resultados: Se detectaron diferencias estadísticamente significativas entre las distintas formulaciones de docetaxel en cuanto a  la incidencia acumulada por ciclo de: modificación de dosis, anemia,  reacciones de hipersensibilidad y anafilaxia, neuropatía, toxicidad palmo- plantar y dermatológica, toxicidad ungueal y edema facial. La  formulación con un menor contenido en impurezas presentó mejores  resultados en modificación de dosis, visitas a urgencias, e incidencia de  anemia y edema facial, pero peores en hospitalización, neutropenia  febril, neuropatía motora y toxicidad palmo-plantar.Conclusiones: Los resultados muestran diferencias en la incidencia de  los eventos adversos de los distintos medicamentos con docetaxel comercializados en nuestro país, con diferencias significativas  entre ellos en algunas de las variables estudiadas. No se ha podido  identificar un medicamento con un mejor perfil de toxicidad. Tampoco se  ha podido establecer su relación con respecto a la composición de  excipientes e impurezas.

Social environment modulates investment in sex trait versus lifespan: red deer produce bigger antlers when facing more rivalry.

Theory predicts that the plastic expression of sex-traits should be modulated not only by their production costs but also by the benefits derived from the presence of rivals and mates, yet there is a paucity of evidence for an adaptive response of sex-trait expression to social environment. We studied antler size, a costly and plastic sex trait, and tooth wear, a trait related to food intake and longevity, in over 4,000 male Iberian red deer (Cervus elaphus hispanicus) from 56 wild populations characterized by two contrasting management practices that affect male age structure and adult sex-ratio. As a consequence, these populations exhibit high and low levels of male-male competition for mating opportunities. We hypothesized that males under conditions of low intra-sexual competition would develop smaller antlers, after controlling for body size and age, than males under conditions of high intra-sexual competition, thus reducing energy demands (i.e. reducing intake and food comminution), and as a consequence, leading to less tooth wear and a concomitant longer potential lifespan. Our results supported these predictions. To reject possible uncontrolled factors that may have occurred in the wild populations, we carried out an experimental design on red deer in captivity, placing males in separate plots with females or with rival males during the period of antler growth. Males living with rivals grew larger antlers than males living in a female environment, which corroborates the results found in the wild populations. As far as we know, these results show, for the first time, the modulation of a sexual trait and its costs on longevity conditional upon the level of intra-sexual competition.

Testosterone and the dark ventral patch of male red deer: the role of the social environment.

The expression of male sexual traits, which is stimulated by testosterone, entails significant costs for individuals. Consequently, natural selection is expected to favour the modulation of sexual trait development according to the balance between its costs and benefits. The proportion of rivals in a population may affect this balance by increasing or decreasing the reproductive benefits associated with the development of sex traits. Here, we explore the relationship between testosterone level and sex trait size under two populational conditions of mate competition: fenced (i.e. high male-male competition; all male age groups are present) and unfenced (i.e. low competition; most males present are juveniles). Our model species is the Iberian red deer (Cervus elaphus hispanicus), and the sex trait is the dark ventral patch that males exhibit during the rutting season. Our results showed that the positive relationship between testosterone levels and the size of the dark ventral patch depends on the environmental level of male-male competition. Only in populations where the operational sex ratio was high (i.e. high proportion of rival males), individuals with high levels of testosterone developed the sex trait. Conversely, when mate competition was low, there was no significant relationship between testosterone level and trait size. This result reinforces the idea that the effect of testosterone in promoting the development of sex traits may be mediated by the intensity of mate competition in the population, as well as the role of sexual selection in the evolution of the dark ventral patch in red deer.

Immune challenge of mating effort: steroid hormone profile, dark ventral patch and parasite burden in relation to intrasexual competition in male Iberian red deer.

Testosterone secretion may regulate the reproductive effort and the development of sexual traits, but it may also involve costs at the immunological and metabolic levels. However, the evidence for this trade-off in wild populations is scarce. Cortisol also plays an important role in mediating the reproductive and immune functions. In this study, we analyzed whether the endoparasite burden relates to hormonal levels (fecal testosterone and cortisol metabolites) and/or morphological sexual traits (size of the dark ventral patch, a trait that indicates reproductive effort in males) in male Iberian red deer. For this purpose, we sampled male red deer harvested during hunting actions in 2 types of populations in south western Spain that differed in structure, affecting the level of male-male competition for mates. We used coprological analyses to estimate the parasite burden mainly of gastrointestinal and bronchopulmonary nematodes and of protozoa, and assessed testosterone and cortisol metabolite levels from fecal pellets. We found a positive relationship of host parasitation with both testosterone levels and the size of the dark ventral patch, but these relationships depended on the intensity of male-male competition in the population, being only found under the high-competition scenario. These results are discussed under the hypothesis of the testosterone immunocompetence handicap, suggesting a cost at the immunological level, and, therefore, higher susceptibility to parasite infection in males that make a greater reproductive effort. However, this effect seems to be modulated by the social environment (male-male competition) that might lead to different optima in testosterone production and sexual trait development.

The intensity of male-male competition may affect chemical scent constituents in the dark ventral patch of male Iberian red deer.

During the mating season, Iberian red deer males (Cervus elaphus hispanicus) present a large visible dark ventral area in their abdomen. This characteristic dark-haired area is formed by the impregnation of the hair with sprayed urine and gland secretions and contains volatile compounds that can be used in intraspecific communication. Here, we used gas chromatography-mass spectrometry (GC-MS) to describe the lipophilic chemicals from the dark ventral patch of males from different populations with different levels of intrasexual competition. Amongst all the compounds found, m-cresol, benzoic acid, cholesterol and 4-hydroxy-benzenopropanoic acid were the most abundant. The proportions of these compounds varied with age as well as with the level of intra-sexual competition, independently of age. In particular, red deer males experiencing higher intra-sexual competition had lower proportions of aromatic compounds (especially m-cresol) but higher proportions of carboxylic acids on their dark bellies. Males in a high male-male competition situation, invest in volatile compounds that can reveal their age, dominance status and condition, and that, in addition, enhance this signal. On the contrary, males from low intra-sexual competition populations have chemical profiles more characteristic of young individuals. This research shows a first glance of how secretion of volatile compounds of male deer can be modulated due to the intensity of male-male competition in the population.

Unprecedented high catecholamine production causing hair pigmentation after urinary excretion in red deer.

Hormones have not been found in concentrations of orders of magnitude higher than nanograms per milliliter. Here, we report urine concentrations of a catecholamine (norepinephrine) ranging from 0.05 to 0.5 g/l, and concentrations of its metabolite DL-3,4-dihydroxyphenyl glycol (DOPEG) ranging from 1.0 to 44.5 g/l, in wild male red deer Cervus elaphus hispanicus after LC-MS analyses. The dark ventral patch of male red deer, a recently described sexually selected signal, contains high amounts of DOPEG (0.9-266.9 mg/l) stuck in the hairs, while DOPEG is not present in non-darkened hair. The formation of this dark patch is explained by the chemical structure of DOPEG, which is a catecholamine-derived o-diphenol susceptible to be oxidized by air and form allomelanins, nitrogen-free pigments similar to cutaneous melanins; by its high concentration in urine; and by the urine spraying behavior of red deer by which urine is spread through the ventral body area. Accordingly, the size of the dark ventral patch was positively correlated with the concentration of DOPEG in urine, which was in turn correlated with DOPEG absorbed in ventral hair. These findings represent catecholamine concentrations about one million higher than those previously reported for any hormone in an organism. This may have favored the evolution of the dark ventral patch of red deer by transferring information on the fighting capacity to rivals and mates. Physiological limits for hormone production in animals are thus considerably higher than previously thought. These results also unveil a novel mechanism of pigmentation based on the self-application of urine over the fur.

A comparative study of contamination in three closed systems for the preparation of hazardous drugs through simulations with fluorescein / Estudio comparativo de contaminación de tres sistemas cerrados para la preparación de fármacos peligrosos mediante simulación con fluoresceína

Objective: The objective of this study was to compare the environmental contamination generated during the preparation of cytostatic agents using three different methods through simulations using fluorescein, and the time required for preparation of each method. Method: A comparative study of the processing of fluorescein mixtures using three types of closed systems was conducted at the centralized unit for hazardous drugs of the Pharmacy Department of a General Teaching Hospital. Environmental contamination was detected in critical points of connection, and in splashes produced at any other points. The main variable was qualitative detection of contamination through ultraviolet light when three methods were compared (method A: ChemoClave(R), method B: SmartSite(R)valve and Texium(R)connector, method C: PhaSealTMwith BD luer extension). A final number of 60 mixtures were prepared to detect differences of at least 5%. Results: Qualitative contamination at the critical points during preparation, was seen in groups A and B for every mixture that was processed. No contamination at all in critical points was seen in any of the mixtures prepared using PhaSealTM. Statistically significant differences were found between arms A and C (p < 0.001) and arms B and C (p < 0.001); no differences were found between arms A and B. Conclusions: The combination of PhaSealTMsystem in conjunction with the BD luer extension for administering hazardous drugs from a tree modality system has been shown to be the system with the lowest level of contamination during processing without increasing the time required for preparation of the mixture

Objetivo: El objetivo de este estudio fue comparar la contaminación ambiental generada durante la preparación de fluoresceína y el tiempo de preparación usando tres sistemas cerrados de transferencia diferentes. Método: Estudio comparativo de elaboración de mezclas de fluoresceína con tres tipos de sistemas cerrados en una unidad de mezclas peligrosas de un Servicio de Farmacia de un Hospital General Universitario. Se consideró contaminación ambiental la detectada en los puntos críticos de conexión y las salpicaduras generadas en cualquier otro punto distinto. La variable principal fue la detección cualitativa mediante luz ultravioleta de la contaminación generada cuando se comparan tres sistemas (sistema A: ChemoClave(R), sistema B: válvula SmartSite(R)y conector Texium(R), sistema C: PhaSealTMcon alargadera luer BD). Se prepararon 60 mezclas para poder detectar diferencias de al menos el 5%. Resultados: Se detectó contaminación en los puntos críticos durante la preparación en todas las mezclas de los grupos A y B. No se detectó contaminación en ninguna de las mezclas en las que se usó el sistema cerrado PhaSealTM. Se encontraron diferencias estadísticas entre los grupos A y C (p < 0,001) y entre los grupos B y C (p < 0,001); no se encontraron diferencias entre los grupos A y B. Conclusiones: La combinación del sistema PhaSealTMy la alargadera luer BD para administrar fármacos peligrosos en la modalidad de árbol ha mostrado ser el sistema con el menor nivel de contaminación durante la preparación, sin que esto se traduzca en aumento en el tiempo de elaboración