Is the step-wise tiered approach for ERA of pharmaceuticals useful for the assessment of cancer therapeutic drugs present in marine environment?

Methotrexate (MTX) and tamoxifen (TMX) cancer therapeutic drugs have been detected within the aquatic environment. Nevertheless, MTX and TMX research is essentially bio-medically orientated, with few studies addressing the question of its toxicity in fresh water organisms, and none to its' effect in the marine environment. To the authors' knowledge, Environmental Risk Assessments (ERA) for pharmaceuticals has mainly been designed for freshwater and terrestrial environments (European Medicines Agency-EMEA guideline, 2006). Therefore, the purpose of this research was (1) to assess effect of MTX and TMX in marine organism using the EMEA guideline, (2) to develop an ERA methodology for marine environment, and (3) to evaluate the suitability of including a biomarker approach in Phase III. To reach these aims, a risk assessment of MTX and TMX was performed following EMEA guideline, including a 2-tier approach during Phase III, applying lysosomal membrane stability (LMS) as a screening biomarker in tier-1 and a battery of biochemical biomarkers in tier-2. Results from Phase II indicated that MTX was not toxic for bacteria, microalgae and sea urchin at the concentrations tested, thus no further assessment was required, while TMX indicated a possible risk. Therefore, Phase III was performed for only TMX. Ruditapes philippinarum were exposed during 14 days to TMX (0.1, 1, 10, 50 µg L(-1)). At the end of the experiment, clams exposed to environmental concentration indicated significant changes in LMS compared to the control (p<0.01); thus a second tier was applied. A significant induction of biomarkers (activity of Ethoxyresorufin O-deethylase [EROD], glutathione S-transferase [GST], glutathione peroxidase [GPX], and lipid peroxidation [LPO] levels) was observed in digestive gland tissues of clams compared with control (p<0.01). Finally, this study indicated that MTX was not toxic at an environmental concentration, whilst TMX was potentially toxic for marine biota. This study has shown the necessity to create specific guidelines in order to evaluate effects of pharmaceuticals in marine environment which includes sensitive endpoints. The inadequacy of current EMEA guideline to predict chemotherapy agents toxicity in Phase II was displayed whilst the usefulness of other tests were demonstrated. The 2-tier approach, applied in Phase III, appears to be suitable for an ERA of cancer therapeutic drugs in the marine environment.

SURGICAL CORRECTION OF TRAUMATIC PATELLAR LUXATION IN AN EURASIAN LYNX (LYNX LYNX).

The aim of this report is to describe the successful surgical repair of a traumatic medial patellar luxation in a 4-yr-old female Eurasian lynx ( Lynx lynx ). The animal presented with hind limb lameness. Physical and radiographic examinations were performed under sedation. After diagnosing a medial patellar luxation, surgical repair was recommended. A combination of soft tissue reconstructive techniques was used to repair the medial patellar luxation. The limb was not immobilized postoperatively, but the animal was confined to a cage for 1 mo. The recovery was uneventful and return to normal activity was observed within 1 mo. Soft tissue reconstructive techniques can be used as the only surgical treatment for the repair of a traumatic patellar luxation in both domestic and wild animals.

Proposed improvements to existing water monitoring systems in automatic radiological warning networks.

The main objective of automatic environmental radiation warning networks is to quantify a set of radiological parameters corresponding to the medium being monitored (water, air, etc) in the shortest possible time so as to be able to provide rapid and precise information on the medium's radiological status, and on any alterations that may occur and their severity. Specifically, in this paper we present the substantial improvements that have been carried out in an automatic near-real-time radiation monitoring of a water system belonging to Radiation Alert Network of Extremadura (RARE) in southwest Spain. These improvements are based on the incorporation of (i) a gamma spectrometry system with solid scintillation detectors and compact digital electronics, (ii) continuous measurement of the water flow that is being monitored, (iii) improvements in the maintenance tasks required to optimise the operation of this type of equipment and (iv) the controlled and automated collection of water samples so that, in the case of a possible radiological anomaly, it will be possible to perform ulterior specific complementary determinations in a low-background laboratory.

Automated FerryBox monitoring reveals the first recorded river induced crude oil seep transport to the Strait of Magellan in southern Patagonia.

This study presents the first documented occurrence of a natural crude oil seep plume associated with river discharge along the Strait of Magellan in southern Patagonia in modern times. Between September and December 2022, hydrocarbon signals were detected using a crude oil sensor integrated into a FerryBox system that traversed the Strait of Magellan and several channels of southern Patagonia, covering approximately 510 km. The highest levels of crude oil signals were observed in the mid-basin of the Strait of Magellan. These signals exhibited a strong negative correlation with sea surface salinity, coinciding with the water discharge from the San Juan River. Notably, during periods of high river discharge, typically exceeding 15 m3 s-1, a distinct crude oil plume was detected moving towards the Magellan Strait. Conversely, when river discharge fell below this threshold, no noticeable crude oil signal was observed. As river discharge decreased and winds intensified during the austral summer, the crude oil signal gradually dissipated. This observation suggests that the dispersion of crude oil becomes limited during periods of low river discharge, as buoyant currents remain confined close to the coast. Historical records indicate that this seep has been releasing hydrocarbons into the Strait of Magellan for at least the past 120 years, implying a long history of chronic crude oil input into this relatively isolated region of the world. This finding shows the potential contribution to the understanding of marine ecosystems dynamics and potential pollutants in poorly studied regions through the use of automated monitoring FerryBox system, enabling both spatial and temporal high-resolution surveys.

A DWI-based hypoxia model shows robustness in an external prostatectomy cohort.

Introduction: Prostate cancer hypoxia is a negative prognostic biomarker. A promising MRI-based tool to assess hypoxia is the 'Consumption and Supply based Hypoxia' (CSH) model based on diffusion-weighted imaging (DWI). The aim of the study was to validate the association between the CSH hypoxia fraction (HFDWI) with pathological Grade Group (pGG) and pathological T-staging (pTstage) in an external prostatectomy cohort. Methods: Apparent diffusion coefficient (ADC) and fractional blood volume (fBV) maps were assessed from DWI data from 291 prostatectomies and combined by the CSH model. HFDWI was calculated for each lesion after median scaling of ADC and fBV to address differences in acquisition and analysis between centers. The absolute HFDWI values and the associations of HFDWI between pGG < 3 versus ≥ 3, and pTstage = 2 versus = 3 in the Netherlands Cancer Institute (NKI) cohort were compared to the obtained by original cohort (Oslo cohort). Statistical T- and Mann-Whitney tests (p<0.05) were performed. Pearson correlation was determined between HFDWI and individual pGG groups. Results: The HFDWI showed comparable absolute values and similar metric performance as in the original published cohort. Higher HFDWI values were observed for higher pGG (Oslo: 0.27; NKI: 0.24) compared to lower pGG (Oslo: 0.11; NKI: 0.17). Similar results were obtained for pTstage. Furthermore, HFDWI demonstrated a significant positive correlation with pGG groups 1-5 (ρ = 0.41, p<0.001). Conclusion: The CSH model exhibited sufficient robustness in the external cohort, suggesting a plausible reflection of true hypoxia and enabling the use of the HFDWI metric for further research into prostate cancer and hypoxia.

Lymphovascular Space Invasion in Early-Stage Endometrial Cancer (LySEC): Patterns of Recurrence and Predictors. A Multicentre Retrospective Cohort Study of the Spain Gynecologic Oncology Group.

The main aim is to compare oncological outcomes and patterns of recurrence of patients with early-stage endometrioid endometrial cancer according to lymphovascular space invasion (LVSI) status. The secondary objective is to determine preoperative predictors of LVSI. We performed a multicenter retrospective cohort study. A total of 3546 women diagnosed with postoperative early-stage (FIGO I-II, 2009) endometrioid endometrial cancer were included. Co-primary endpoints were disease-free survival (DFS), overall survival (OS), and pattern of recurrence. Cox proportional hazard models were used for time-to-event analysis. Univariate and multivariate logistical regression models were employed. Positive LVSI was identified in 528 patients (14.6%) and was an independent prognostic factor for DFS (HR 1.8), OS (HR 2.1) and distant recurrences (HR 2.37). Distant recurrences were more frequent in patients with positive LVSI (78.2% vs. 61.3%, p < 0.01). Deep myometrial invasion (OR 3.04), high-grade tumors (OR 2.54), cervical stroma invasion (OR 2.01), and tumor diameter ≥ 2 cm (OR 2.03) were independent predictors of LVSI. In conclusion, in these patients, LVSI is an independent risk factor for shorter DFS and OS, and distant recurrence, but not for local recurrence. Deep myometrial invasion, cervical stroma invasion, high-grade tumors, and a tumor diameter ≥ 2 cm are independent predictors of LVSI.

HLA in Jaidukama: an Amerindian secluded Colombian population with new haplotypes and Asian and Pacific-shared alleles.

America first inhabitants and peopling are still debated. In order to increase knowledge about these questions, we have aimed to detect HLA genes of an Amerindian secluded community: Jaidukama, who lives in North Colombia Equatorial forest. HLA genotyping and extended haplotype calculations were carried out in 39 healthy individuals belonging to 13 families. HLA frequencies were compared to other Amerindians and worldwide populations by calculating genetic distances, relatedness dendrograms and correspondence analyses. Only four DRB1 alleles were found (*0404, *0407, *1402 and *1602); however a total of 17 Amerindian different extended class I-class II HLA haplotypes were directly counted from the family studies, nine of them were specific of Jaidukamas. Some of the alleles or group of alleles within an extended haplotype (i.e. DQB1-DRB1) were also found in Asians and Pacific Islanders, further supporting existence of Asian and Pacific gene flow with Amerindians or a common founder effect. It is further supported that HLA extended haplotypes vary faster than alleles in populations. It is concluded that this unique model of Amerindian secluded families study suggests that rapid HLA haplotype variation may be more important than allele variation for survival (starting immune responses). This work may also be useful for future transplant programs in the area.

Expression and function of HOXA genes in normal and neoplastic ovarian epithelial cells.

We studied the roles of three HOXA genes in cultured normal ovarian surface epithelial (OSE) cells and ovarian cancer cells. They included HOXA4 and HOXA7 because, by cDNA microarray analysis, these were more highly expressed in invasive ovarian carcinomas than in benign or borderline (noninvasive) ovarian tumors, and HOXA9 because it characterizes normal oviductal epithelium, which resembles ovarian serous adenocarcinomas. The three HOXA genes were more highly expressed when OSE cells were dividing and motile than when they were confluent and stationary, and also when they dispersed in response to EGF treatment or to reduced calcium concentrations in culture media. The expression of the HOXA genes varied among ovarian cancer cell lines, but was highest in lines with compact epithelial morphologies. We focused on HOXA4 as the most highly expressed in the ovarian carcinoma array. HOXA4 expression did not parallel proliferative activities of either OSE or ovarian cancer lines. Moreover, modifying HOXA4 expression in ovarian cancer cell lines did not alter either E-cadherin expression or CA125 secretion. However, HOXA4 downregulation enhanced EGFR phosphorylation and migration in serum-starved OSE and ovarian cancer cells in response to EGF, and enhanced migration of all ovarian cancer lines in 5% serum even without EGF treatment. Thus, HOXA4 expression does not correlate with proliferation or with epithelial differentiation, but it increases in response to OSE cell dispersion and negatively regulates EGFR activation and the motility of OSE and of ovarian cancer cells. HOXA4 expression was highest in cancer lines with compact epithelial growth patterns, suggesting, again, an anti-dispersion function. In summary, increased HOXA4 expression in ovarian cancer appears to constitute a tumor-suppressive, homeostatic response to aberrant cell behavior, and, in particular, to cell dispersion and migration.

Improved sea-urchin embryo bioassay for in situ evaluation of dredged material.

Sediments usually contain many contaminants derived from human activities. In case of dredging activities, these sediment-bound contaminants arise following the excavation and remobilization of sediments. Previous studies have used different species of clam, crabs, lugworms, etc. for the evaluation of dredged material in situ, but there are not studies that use acute bioassays for these purposes. The sea-urchin embryo bioassay has been chosen to characterize biological effects in situ in two ports of the southwest of Spain, the Port of Huelva and the Port of Cadiz. The sea-urchin embryo bioassay has been adapted for in situ evaluation of seawater quality in coastal areas, however, they are necessary for further improvements to take into account differences of temperature between sites. This temperature variation is one of the principal reasons (other than pollution) of larval mortality and the slow down in the growth rate of the urchin. In the present study a bioassay was conducted in both field and laboratory conditions, in order to compare the effects in situ with the effects under controlled conditions of temperature, salinity and oxygen dissolved. Results showed a good correlation between samples obtained in situ and in the laboratory, but in the field the percentage of normal pluteus larvae is less than under laboratory conditions.

Política industrial farmacéutica, un requisito clave para la autonomía sanitaria de Colombia / Pharmaceutical industrial policy, a key requirement for health autonomy in Colombia

Colombia depende de la importación de medicamentos, así como de gran parte de los materiales (principios activos y excipientes) requeridos para su elaboración; problemática que genera consecuencias sanitarias y macroeconómicas, las cuales se agudizan en el contexto de desindustrialización nacional y de disrupción tecnológica. De esta manera, se acepta que la disponibilidad y acceso a medicamentos y otras tecnologías sanitarias esenciales son un requisito fundamental para alcanzar la autonomía sanitaria de un país. Por lo tanto, resulta imprescindible coordinar esfuerzos entre diversos sectores sociales para desarrollar una agenda pública enfocada a la creación de condiciones que fortalezcan las capacidades científicas y tecnológicas de la industria farmacéutica local, y con ello, mejorar el suministro farmacéutico del país. En el presente documento se presentan conceptos teóricos y prácticos que deberían ser considerados en la definición y materialización de una política pública encaminada a fortalecer la industria farmacéutica y favorecer la autonomía sanitaria de Colombia.

Colombia has a notorious dependency on the importation of medicines, as well as a large part of the materials (active ingredients and excipients) required for their manufacture. This problem generates health and macroeconomic consequences, which are exacerbated in the context of national deindustrialization and technological disruption. In this way, it is accepted that the availability and access to medicines and other essential health technologies are a fundamental requirement to achieve the health autonomy of a country. Therefore, it is crucial to coordinate efforts between several social sectors to develop a public agenda focused on creating conditions that allow strengthening the scientific and technological capabilities of the local pharmaceutical industry, thereby, improving the country's pharmaceutical supply. This document presents conceptual and practical topics that should be considered to defining and materializing a public policy aimed at strengthening the local pharmaceutical industry and favoring Colombia's sanitary autonomy.

Pathology findings and clinical outcomes after risk reduction salpingo-oophorectomy in BRCA mutation carriers: a multicenter Spanish study.

OBJECTIVE: To determine the incidence of serous tubal intraepithelial carcinoma (STIC) after risk reduction salpingo-oophorectomy(RRSO), and to describe oncological outcomes after RRSO. MATERIALS AND METHODS: BRCA pathogenic mutation carriers who had undergone an RRSO were evaluated in this retrospective multicenter observational study. Patients were only included when fallopian tubes were analyzed following the protocol for Sectioning and Extensively Examining the FIMbria (SEE-FIM). Surgeries were performed between June 2010 and April 2017 at eight Spanish hospitals. RESULTS: A total of 359 patients met the inclusion criteria. STIC was diagnosed in 3 (0.8%) patients; one of them underwent surgical staging due to positive peritoneal washing, with absence of disease at the final pathology report. None of the three patients received adjuvant chemotherapy and were free of disease at last follow-up. Fallopian tube and ovarian carcinoma were diagnosed in 5 (1.4%) and 1 (0.3%), respectively. At a median (range) follow-up time of 29 (3-92) months, five patients had a newly diagnosed breast cancer. Other types of cancer, which were diagnosed during the follow-up time, included: serous primary peritoneal carcinoma (n = 1), serous endometrial carcinoma (n = 1), colon (n = 1), pancreas (n = 1), jaw (n = 1), and lymphoma (n = 1). Seven patients died due to different types of cancer: breast (n = 4), pancreas (n = 1), jaw (n = 1), and colon (n = 1). CONCLUSION: The incidence of STIC after RRSO in BRCA mutation carriers is low (0.8%) and it presents an excellent oncological outcome. Patients after RRSO, however, run the risk to develop other types of cancer during follow-up and should be properly advised before the prophylactic surgery.

Removal naturally occurring radionuclides from drinking water using a filter specifically designed for Drinking Water Treatment Plants.

The occurrence of naturally occurring radionuclides in drinking water can pose health hazards in some populations, especially taking into account that routine procedures in Drinking Water Treatment Plants (DWTPs) are normally unable to remove them efficiently from drinking water. In fact, these procedures are practically transparent to them, and in particular to radium. In this paper, the characterization and capabilities of a patented filter designed to remove radium from drinking water with high efficiency is described. This filter is based on a sandwich structure of silica and green sand, with a natural high content manganese oxide. Both sands are authorized by Spanish authorities to be used in Drinking Water Treatment Plants. The Mn distribution in the green sand was found to be homogenous, thus providing a great number of adsorption sites for radium. Kinetic studies showed that the 226Ra adsorption on green sand was influenced by the content of major cations solved in the treated water, but the saturation level, about 96-99%, was not affected by it. The physico-chemical parameters of the treated water were unaltered by the filter. The efficiency of the filter for the removal of 226Ra remained unchanged with large water volumes passed through it, proving its potential use in DWTP. This filter was also able to remove initially the uranium content due to the presence of Fe2O3 particles in it, although it is saturated faster than radium.

Diurnal preference, mood and the response to morning light in relation to polymorphisms in the human clock gene PER3.

PER3 gene polymorphisms have been associated with differences in human sleep-wake phenotypes, and sensitivity to light. The aims of this study were to assess: i) the frequency of allelic variants at two PER3 polymorphic sites (rs57875989 length polymorphism: PER3 4, PER3 5; rs228697 SNP: PER3 C, PER3 G) in relation to sleep-wake timing; ii) the effect of morning light on behavioural/circadian variables in PER3 4 /PER3 4 and PER3 5 /PER3 5 homozygotes. 786 Caucasian subjects living in Northern Italy donated buccal DNA and completed diurnal preference, sleep quality/timing and sleepiness/mood questionnaires. 19 PER3 4 /PER3 4 and 11 PER3 5 /PER3 5 homozygotes underwent morning light administration, whilst monitoring sleep-wake patterns and the urinary 6-sulphatoxymelatonin (aMT6s) rhythm. No significant relationship was observed between the length polymorphism and diurnal preference. By contrast, a significant association was observed between the PER3 G variant and morningness (OR = 2.10), and between the PER3 G-PER3 4 haplotype and morningness (OR = 2.19), for which a mechanistic hypothesis is suggested. No significant differences were observed in sleep timing/aMT6s rhythms between PER3 5 /PER3 5 and PER3 4 /PER3 4 subjects at baseline. After light administration, PER3 4 /PER3 4 subjects advanced their aMT6s acrophase (p < 0.05), and showed a trend of advanced sleep-wake timing. In conclusion, significant associations were observed between PER3 polymorphic variants/their combinations and both diurnal preference and the response to light.

[Effectiveness of treatment with tiotropium in patients who suffer from COPD]. / Efectividad del tratamiento con bromuro de tiotropio en pacientes con enfermedad pulmonar obstructiva crónica.

BACKGROUND AND OBJECTIVE: We assessed the clinical and functional changes taking place in patients diagnosed of chronic obstructive pulmonary disease (COPD) who have been treated with ipratropium bromide, three months after this medicament was replaced by the new tiotropium bromide. PATIENTS AND METHOD: A prospective intervention survey was carried out in a primary health-care area in patients who suffered from COPD who fulfilled the inclusion criteria. Before changing the treatment and three months after changing it, we carried out the following procedures: spirometry, 6 minutes walking test (6MWT) with pulsioximetry before and after the exercise, St. George Respiratory Questionnaire (SGRQ) and assessment of: patient's chronic dyspnea, degree of patient's compliance, adverse effects and degree of satisfaction with the new drug. RESULTS: 24 patients (22 males and 2 females) with a mean age of 68.54 years participated in the survey. The FVC improved a 4.92% (120 ml) [p = .27] and the FEV1, 14.16% (170 ml) [p < .001]. The degree of compliance arose from 67.54% to 96.73% (p < .001), the degree of dyspnea decreased from 4.63 to 3.89 (p = .141), the 6MWT increased in 23.79 m (p = .027) and the global mark of the SGRQ reduced (improved) in 13.35 points. There were adverse effects in 5 cases (mouth dryness in 4 patients and headache in 1 of them) and 15 patients (62.5%) said the new drug was better. CONCLUSIONS: Most analyzed parameters had positive changes three months after changing the treatment. Tiotropium may be a valid alternative in the treatment of patients suffering from COPD in a stable stage.

The self-morningness/eveningness (Self-ME): An extremely concise and totally subjective assessment of diurnal preference.

The assessment of diurnal preference, or the preferred timing of sleep and activity, is generally based on comprehensive questionnaires such as the Horne-Östberg (HÖ). The aim of the present study was to assess the reliability of a subject's self-classification as extremely morning (Self-MM), more morning than evening (Self-M), more evening than morning (Self-E) or extremely evening (Self-EE) type, based on the last question of the HÖ (Self-ME). A convenience sample of 461 subjects [23.8 ± 4.7 years; 322 females] completed a full sleep-wake assessment, including diurnal preference (HÖ), night sleep quality (Pittsburgh Sleep Quality Index, PSQI), daytime sleepiness (Karolinska Sleepiness Scale, KSS), and habitual sleep-wake timing (12 d sleep diaries; n = 296). Significant differences in HÖ total score were observed between Self-ME classes, with each class being significantly different from neighboring classes (p < 0.0001). Significant differences in sleep-wake timing (bed time, try to sleep and sleep onset, wake up, and get up time) were observed between Self-ME classes. Such differences were maintained when sleep-wake habits were analysed separately on work and free days, and also in a smaller group of 67 subjects who completed the Self-ME as a stand-alone rather than as part of the original questionnaire. Significant differences were observed in the time-course of subjective sleepiness by Self-ME class in both the large and the small group, with Self-MM and Self-M subjects being significantly more alert in the morning and sleepier in the evening hours compared with their Self-E and Self-EE counterparts. Finally, significant differences were observed in night sleep quality between Self-ME classes, with Self-EE/Self-E subjects sleeping worse than their Self-MM/Self-M counterparts, and averaging just over the abnormality PSQI threshold of 5. In conclusion, young, healthy adults can define their diurnal preference based on a single question (Self-ME) in a way that reflects their sleep-wake timing, their sleepiness levels over the daytime hours, and their night sleep quality. Validation of the Self-ME across the decades and in diseased populations seems worthy.

Are standard tests sensitive enough to evaluate effects of human pharmaceuticals in aquatic biota? Facing changes in research approaches when performing risk assessment of drugs.

Nowadays, the presence of pharmaceutical products in aquatic environments is not only common, but is also of significant concern regarding the adverse effect they may produce to aquatic biota. In order to determine the adverse effects of caffeine (CAF), ibuprofen (IBU), carbamazepine (CBZ) and novobiocin (NOV), at environmental occurring concentrations, standardized endpoints applied in current guidelines were evaluated in four organisms including bioluminescence response in Vibrio fischeri, growth inhibition in Isochrysis galbana (marine water) and Pseudokirchneriella subcapitata (fresh water) and fertilization and embryo-larval development in Paracentrotus lividus. To reach this aim bioassays were implemented by exposing organisms to water spiked with drugs dissolved in DMSO (0.001% v/v). Risk characterization was performed, calculating the environmental impact of drugs by calculating environmental concentration and predicted no effect concentration ratio (MEC/PNEC). Results indicate that acute toxicity was found above environmental concentrations in the order of mg L(-1) for bacteria bioluminescence, microalgae growth inhibition and sea urchin fertilization. However, teratogenicity was observed on sea urchin after exposure to environmental concentrations of drugs at 0.00001 mg L(-1); at this concentration CBZ and IBU were found to reduce significantly the embryo-larval development compared to controls (p<0.01). The risk calculated for selected drugs suggested they are harmless for aquatic environment except when applying the embryo-larval development endpoint. Endpoints applied in this study showed the necessity of using more sensitive responses, when assessing risk of pharmaceuticals in aquatic environments, since endpoints applied in current guidelines may not be suitable.