Advanced practice in radiotherapy across Europe: stakeholders' perceptions of implementation and evolution.

INTRODUCTION: Adapting radiotherapy services with workforce innovation using skills-mix or task-shifting optimises resources, supporting current and future demands. Advanced practitioners (APs) work at a different level of practice (beyond initial registration) across four pillars: clinical practice, leadership and management, education, and research. There is limited cross-country research on the advanced therapeutic radiographers/radiation therapists (TR/RTTs), particularly in Europe. This study aimed to investigate European radiotherapy stakeholders' perceptions regarding current and future advanced practice (AP). METHODS: From June to September 2022, one-to-one online semi-structured interviews were conducted in English, and audio and video were recorded. Full verbatim audio files were independently transcribed and checked by interviewer and interviewees. Braun and Clarke's seven steps guided the thematic analysis (using NVivo). RESULTS: Thirty-three interviewees working or studying in 16 European countries represented practitioners (n=14), managers (n=6), educators (n=4), professional bodies (n=4), students (n=3), and regulators (n=2). Four overarching themes emerged: "AP drivers and outcomes", "AP challenges vs enablers", "Current vs future AP", "Becoming and being advanced practitioner". Participants identified research as the neglected AP pillar due to a lack of protected time, limited staff skills, no research culture, no funding, workload, and clinical priorities. Interviewees highlighted the importance of consistency in job titles, harmonisation of education models and curricula, definition of AP requirements, and support for all AP pillars through job plans and workforce planning. CONCLUSION: Neither the profession nor education of TR/RTTs are harmonised across Europe, which is highly reflected in advanced-level practice. Advanced TR/RTTs should work across all pillars, including research, and these should be embedded in master's programmes, including leadership. IMPLICATIONS FOR PRACTICE: This study highlights a policy gap in the education and practice of APs in radiotherapy.

High resilience of campos rupestres plants to the interaction of drought and fire.

The concurrent impacts of multiple disturbances have the potential to modify ecosystem functioning by diminishing recovery capacity and resilience. Nevertheless, it remains uncertain how plant species from tropical communities respond to the cumulative effects of drought and fire. In this study, we evaluated the responses of six plant species from campos rupestres subjected to a mild drought followed by fire and tested if plants subjected to simulated drought show reductions in carbon uptake and depletion of non-structural carbohydrate (NSC) reserves, thus constraining their resprouting. We monitored monthly variations in leaf gas exchange and aboveground biomass over 18 months. Subsequently, an accidental fire occurred in the study area, leading us to collect samples of belowground structures for NSC analyses on the day of the burn. There were no differences in the frequency of resprouting between the above two conditions. Additionally, gas exchange in most species either remained stable or increased after the fire. Drought had no adverse effects on NSC reserves in the belowground structures and may have contributed to species resprouting after fire. The impact of drought pre-conditions on post-fire aboveground biomass was generally minor for most species, except Vellozia nivea, which displayed roughly a 5% reduction in biomass following the drought. Our findings highlight the remarkable resilience of campos rupestres species, even after enduring 18 months of reduced water availability and an unintended fire event. These species demonstrated the capacity to maintain their physiological functions and resprouting capacities after a fire event, underscoring their strong recovery potential.

Patient-reported perspectives of therapeutic radiographers when undergoing radiotherapy: A European multi-centre study.

INTRODUCTION: Radiotherapy is delivered almost exclusively by therapeutic radiographers/radiation therapist (RTTs). Patient's perspectives of RTTs affect levels of trust and confidence in the profession and can have a significant impact on overall radiotherapy experience. The study reports patients' perspectives of RTTs from their experience of undergoing radiotherapy. Four partner sites collaborated in this research and included Malta, Poland, Portugal, and the UK (lead site). METHODS: A survey was developed to gather information from patients receiving radiotherapy or who had had radiotherapy within the previous 24 months. Participants ranked their responses to 23 statements relating to person-centred care on a 5-point scale of 1 (strongly disagree) to 5 (strongly agree). Mann-Whitney or Kruskal Wallis tests were applied to test differences in responses to 5 key statements for patient characteristics including gender, age group, diagnosis, country, time spent with RTTs and number of fractions remaining at survey completion. RESULTS: Three hundred and forty-seven surveys are included. Patients report a positive perception of RTTs (95.4% agree with 'I feel cared for'). Statistically significant differences in responses were found between gender, diagnosis, country, time spent with RTTs and fractions of radiotherapy remaining. Patients who had more time with RTTs and completed their surveys during radiotherapy had a more positive perception of RTTs. CONCLUSION: This study suggests that sufficient time with RTTs is key to ensuring a positive radiotherapy patient experience. RTTs being attentive, understanding, and informative are most predictive of a positive overall patient experience. Timing of survey completion can influence responses. IMPLICATIONS FOR PRACTICE: RTT education programmes should incorporate training on person-centred care at all levels. Further research into patient experience of RTTs is warranted.

An investigation of digital skills of therapeutic radiographers/radiation therapists: A european survey of proficiency level and future educational needs.

INTRODUCTION: This study aims to assess the proficiency level of digital skills, the factors influencing that level and the training needs of Therapeutic Radiographers/Radiation Therapists (TR/RTTs), due to the differences in technology availability and accessibility, variations in the regulation and education of TR/RTTs in European countries, and the lack of a digital skills framework. METHODS: An online survey was distributed to TR/RTTs working in Europe to capture their self-assessment of proficiency levels of digital skills when performing their clinical role. Information was also gathered regarding training, work experience and level of information and communication technology (ICT) skills. Quantitative measures were analysed using descriptive statistics and correlation between variables, and qualitative responses using thematic analysis. RESULTS: 101 respondents from 13 European countries completed the survey. Digital skills in treatment planning followed by management and research were the least developed skills, while the most developed were transversal digital skills followed by digital skills in treatment delivery. The Radiotherapy areas of practice where TR/RTT has experience (e.g. Planning Image, Treatment Planning, Treatment), as well as the level of generic ICT skills (communication, content creation and problem-solving), was related to the level of proficiency of TR/RTT digital skills. Greater scope of practice and level of generic ICT were associated with a higher level of TR/RTT digital skills. Thematic analysis allowed the identification of new sub-themes to be included in the training of TR/RTTs. CONCLUSION: Education and training of TR/RTTs should be improved and adapted to the current needs of digitalisation to avoid differences in digital proficiency levels. IMPLICATIONS FOR PRACTICE: Aligning TR/RTTs' digital skill sets with emerging digitalisation will improve current practice and ensure the best care to all RT patients.

Patients' perspectives of the skills and competencies of therapy radiographers/radiation therapists (TRs/RTTs) in the UK, Portugal and Malta; a qualitative study from the SAFE Europe project.

INTRODUCTION: The role of the Therapy Radiographer/Radiation Therapist (TR/RTT) is to provide radiotherapy to patients with a cancer diagnosis. This includes, not only administration of treatment, but also management of side-effects and provision of support/care. Despite this role being consistent throughout Europe, there is currently no standardisation of education for TRs/RTTs. The SAFE EUROPE project aims to standardize TR/RTT education to enable 'safe and free exchange' of TRs/RTTs across Europe. Consequently, this study aims to explore patients' perspectives regarding the current skills and competencies of TRs/RTTs. METHODS: From May 2021 to February 2022, semi-structured interviews were conducted with patients who had recently received radiotherapy in the UK, Malta and Portugal. Ethical approval for this study was granted by the NHS Research Ethics Committee with additional local approvals obtained. RESULTS: Forty-eight participants from the UK (n = 18), Portugal (n = 19), and Malta (n = 11) completed interviews. Participants described high satisfaction with TRs'/RTTs' competence and skills in all three countries. The main theme arising from the analysis was the importance of trust building with TRs/RTTs. Six factors were identified as influencing levels of trust: communication; side-effect management; team consistency; relational skills; patient dignity; and competence. A small number of patients reported feeling rushed and not having their physical and emotional needs met by TRs/RTTs. CONCLUSION: This multicentre study demonstrated that patients perceive TRs/RTTs in the UK, Malta and Portugal as highly competent and skilled. Practical recommendations are provided to address identified deficits in practice, which can be addressed through adaptation of TR/RTT education/training and clinical practice. IMPLICATIONS FOR PRACTICE: Recommendations arising from this study are important to ensure that TRs/RTTs have transferable skills that provide consistently high quality care to patients throughout Europe.

Advanced practice roles amongst therapeutic radiographers/radiation therapists: A European survey.

INTRODUCTION: Advanced Practice (AP) roles in Radiotherapy (RT) over time are variable, often locally developed and not underpinned by professional standards which leads to conceptual and practical gaps. This study aimed to assess AP roles amongst Therapeutic Radiographers/Radiation Therapists (TR/RTTs) and identify educational gaps for this level across Europe. METHODS: An anonymous online survey was designed, validated, and distributed across Europe. Convenience sampling was used to recruit advanced TR/RTTs practitioners or TR/RTTs working in AP roles. Descriptive analysis from closed questions and thematic analyses from open questions are reported. RESULTS: A total of 272 responses were obtained, of which 189 eligible participations were from 21 European countries. 42% of respondents acknowledged additional education required to perform AP, and 25% reported a minimum of five years of RT practice to perform AP roles/tasks. There is a trend to work more on the clinical practice domain with a low percentage of working time allocated to research. Inconsistency was found in job titles, scopes of practice, and educational backgrounds across and even within countries. Education needs regarding knowledge about image-guided and adaptive RT, multimodal imaging and technologies, and advanced treatment planning were found. Training needs on leadership and management skills and clinical site-specific expertise were identified. CONCLUSION: This study clearly shows a gap in education support, a need for standardisation in job titles and scopes of practice across Europe. IMPLICATIONS FOR PRACTICE: As the first large-scale assessment of current AP roles and educational support amongst TR/RTTs across Europe, this study recommends the establishment of governance structure and role regulation. It also informs the curricula for master programmes to align the education with current and future practice.

Digital skills of therapeutic radiographers/radiation therapists - Document analysis for a European educational curriculum.

INTRODUCTION: It is estimated that around 50% of cancer patients require Radiotherapy (RT) at some point during their treatment, hence Therapeutic Radiographers/Radiation Therapists (TR/RTTs) have a key role to play in patient management. It is essential for TR/RTTs to keep abreast with new technologies and continuously develop the digital skills necessary for safe RT practice. The RT profession and education is not regulated at European Union level, which leads to heterogeneity in the skills developed and practised among countries. This study aimed to explore the white and grey literature to collate data on the relevant digital skills required for TR/RTTs practice. METHODS: An exhaustive systematic search was conducted to identify literature discussing digital skills of TR/RTTs; relevant grey literature was also identified. A thematic analysis was performed to identify and organise these skills into themes and sub-themes. RESULTS: 195 digital skills were identified, organised in 35 sub-themes and grouped into six main themes: (i) Transversal Digital Skills, (ii) RT Planning Image, (iii) RT Treatment Planning, (iv) RT Treatment Administration, (v) Quality, Safety and Risk Management, and (vi) Management, Education and Research. CONCLUSION: This list can be used as a reference to close current gaps in knowledge or skills of TR/RTTs while anticipating future needs regarding the rapid development of new technologies (such as Artificial Intelligence or Big Data). IMPLICATIONS FOR PRACTICE: It is imperative to align education with current and future RT practice to ensure that all RT patients receive the best care. Filling the gaps in TR/RTTs skill sets will improve current practice and provide TR/RTTs with the support needed to develop more advanced skills.

Advanced practice roles of therapeutic radiographers/radiation therapists: A systematic literature review.

INTRODUCTION: Advances in Radiotherapy (RT) technology and increase of complexity in cancer care have enabled the implementation of new treatment techniques. Subsequently, a greater level of autonomy, responsibility, and accountability in the practice of Therapeutic Radiographers/Radiation Therapists (TR/RTTs) has led to Advanced Practice (AP) roles. The published evidence of this role is scattered with confusing terminology and divergence regarding the perception of whether a specific role represents AP internationally. This study aims to establish an international baseline of evidence on AP roles in RT to identify roles and activities performed by TR/RTTs at advanced level practice and to summarise the impact. METHODS: A systematic PRISMA review of the literature was undertaken. Thematic analysis was used to synthesise the roles and associated activities. Six RT external experts validated the list. The impact was scrutinised in terms of clinical, organisational, and professional outcomes. RESULTS: Studies (n = 87) were included and categorised into four groups. AP roles were listed by clinical area, site-specific, and scope of practice, and advanced activities were organised into seven dimensions and 27 sub-dimensions. Three most-reported outcomes were: enhanced service capacity, higher patient satisfaction, and safety maintenance. CONCLUSION: Evidence-based AP amongst TR/RTTs show how AP roles were conceptualised, implemented, and evaluated. Congruence studies have shown that TR/RTTs are at par with the gold-standard across the various AP roles. IMPLICATIONS FOR PRACTICE: This is the first systematic literature review synthetisising AP roles and activities of TR/RTTs. This study also identified the main areas of AP that can be used to develop professional frameworks and education guiding policy by professional bodies, educators and other stakeholders.

Microencapsulated and uncoated butyric acid as alternative additives to the regeneration of intestinal mucosa in broilers challenged with Eimeria spp.

1. The effect of microencapsulated and uncoated butyric acid as an alternative to antibiotics on performance, intestinal morphology and regeneration of intestinal mucosa was studied in birds experimentally infected with Eimeria spp. 1 to 42 d-old.2. A total of 1,320 male Cobb® broiler chicks were allocated to one of five treatments in a completely randomised design, comprising a negative control, uncoated butyric acid (UA), microencapsulated butyric acid (MA), combined U + M butyric acid and a positive control (antibiotic+anticoccidial) in six replications. At 16 d-old, the birds were inoculated orally with 0.5 ml of a solution containing an Eimeria spp. pool.3. At 21 d of age, the birds receiving butyric acid alone had higher body weight gain (BWG) and feed intake (FI) compared to those supplemented with the blend of acids. For the total rearing period, in all variables, the positive control performed best (P < 0.001).4. At 14 d of age, birds that received diets containing UA had a deeper crypt depth in the jejunum than those fed diets containing microencapsulated acid (P = 0.0194). At 21 d of age, the birds fed the acids had higher villi (P = 0.0058) in the duodenum, compared to the negative control group.5. Supplementation with microencapsulated acid contributed to the intestinal health and recovery of post-challenge birds, but did not result in improvements in performance.

Agaricus subrufescens and Pleurotus ostreatus mushrooms as alternative additives to antibiotics in diets for broilers challenged with Eimeria spp.

1. The effect of A. subrufescens and P. ostreatus mushrooms as an alternative to antibiotics (avilamycin or monensin sodium) on performance, intestinal morphometry, immunity, and biochemical profile of broilers challenged with Eimeria spp. was studied from 1 to 42 d old. A total of 900 male Cobb® broiler chicks were distributed, according to a completely randomised design, into five treatments with six replicates each.2. The treatments consisted of: negative control (NC) - basal diet (BD) with no anticoccidial or antibiotic (non-challenged birds); negative control challenged (NCC) - NC fed to Eimeria spp. challenged birds; BD with 0.2% A. subrufescens inclusion for challenged birds (As), BD with 0.2% P. ostreatus inclusion for challenged birds (Po); and a positive control - BD with anticoccidial and antibiotic inclusion for challenged birds (ATB).3. At 11 d.o., the birds were each inoculated orally with 1 ml solution containing 2 × 105 sporulated oocysts/ml Eimeria acervulina and 2 × 104 sporulated oocysts/ml E. maxima and E. tenella.4. Birds subjected to Eimeria spp. challenge up to 21 d of age had greater crypt depth, indicating that the presence of undesirable microorganisms had an effect on cell proliferation.5. At 21 d old, the birds receiving ATB had higher average weight gain (AWG), feed intake (AFI), and feed conversion ratio (FCR) compared to those fed diets supplemented with mushrooms (As or Po). For the total rearing period (42 days), the birds that received ATB had higher AWG and AFI (P < 0.001) compared to those that received As or Po diets. Feeding avilamycin did not affect (P = 0.0676) FCR compared to the As or Po diet groups.6. From the morphometric and blood analyses there were no differences between broilers fed ATB, Po or As diets in either rearing periods. However, Po and As supplementation lowered blood triglyceride levels. At 21d there was a difference (P < 0.05) for MCV and haemoglobin, in which the mushrooms were similar to the antibiotic. At 42 d, there was a difference (P < 0.05) in haematocrit, erythrocyte, MCV, H: L, protein and albumin variables, in which the use of mushrooms was similar to the positive control, demonstrating that both (mushrooms and antibiotics) promoted a certain improvement in the health of the chickens.7. A. subrufescens and P. ostreatus can be used in broiler diets without compromising intestinal or haematological status, however, these ingredients did not result in improvements in performance.

Prognostic values of physical and hematological parameters of dogs naturally infected with parvovirus PVC-2: retrospective study of 103 cases / [Valores prognósticos de parâmetros físicos e hematológicos de cães naturalmente infectados pelo parvovírus PVC-2: estudo retrospectivo de 103 casos]

Canine parvovirosis is a high mortality disease with acute clinical picture. However, there are few available resources to help stablish prognosis accurately. This study aimed to determine the prognostic threshold values for vital and hematological parameters of dogs naturally infected by the Carnivore protoparvovirus 1 (CPV). A retrospective study of 103 canine parvovirosis cases was carried out. Twenty seven percent of these (28/103) died, 96% (27/28) of which within the first four days of hospitalization. Deceased animals had significantly higher median values for heart (HR) and respiratory (f) rates, as well as significantly lower systolic blood pressure (SBP) than survivors. Severely leukopenic animals (<1,000 cells/µL), had a significantly higher mortality rate (68%, n=13) compared to that of other patients (P<0.0007). Animals with at least two of the following findings: severe hypotension (SBP< 90mmHg), tachycardia (HR > 150 bpm) and leukopenia, represented 34% (34/101) of the cases and had a survival rate of 29% (10/34), while animals with at most one of these parameters represented 66% (67/101) and had a survival rate of 94% (63/67). The presence of two or three abnormal parameters was significantly related to the higher death risk among dogs with parvovirosis (P<0.0001).(AU)

A parvovirose canina é uma doença de alta mortalidade e de quadro clínico agudo. No entanto, existem poucos recursos para se estabelecer prognóstico de maneira precisa. Este estudo objetivou analisar os valores prognósticos de parâmetros físicos e hematológicos de cães naturalmente infectados pelo Carnivore protoparvovirus 1 (CPV). Um estudo retrospectivo de 103 casos de parvovirose canina foi realizado. Desses, 27% dos animais (28/103) foram a óbito, sendo 96% (27/28) com ocorrência nos primeiros quatro dias de internamento. Os cães que foram a óbito apresentaram medianas das frequências cardíaca (FC) e respiratória (f) significativamente maiores e pressão arterial sistólica (PAS) consideravelmente menor que a dos sobreviventes. Entre os animais mais intensamente leucopênicos (<1.000 células/(L), a taxa de mortalidade (68%, n=13) foi expressivamente maior que a dos demais pacientes (P<0,0007). Os animais com hipotensão grave (PAS<90mmHg), taquicardia (FC>150bpm) e leucopenia intensa (leucometria<1.000 células/µL), ou duas dessas alterações clínicas, representaram 34% (34/101) dos casos e tiveram taxa de sobrevida de 29% (10/34), enquanto os animais com, no máximo, um desses parâmetros alterados representaram 66% (67/101) dos animais, com taxa de sobrevida de 94% (63/67). A presença de dois ou três parâmetros alterados esteve significativamente relacionada ao maior risco de óbito de cães com parvovirose (P<0,0001).(AU)

Genotoxicity effects on Geophagus brasiliensis fish exposed to Doce River water after the environmental disaster in the city of Mariana, MG, Brazil / Efeitos da genotoxicidade em peixes da espécie Geophagus brasiliensis expostos a água do Rio Doce após o desastre ambiental na cidade de Marina, MG, Brasil

Abstract The Doce River basin has suffered the largest environmental accident ever occurred in Brazil with the influx of tailings from Fundão and Santarém, belonging to Samarco mining company, due to the disaster in Mariana. A spill between 50 and 60 million m3 of tailings was estimated by the company. According to Samarco, the wastewater was composed mainly of clay, silt and heavy metals like iron, copper and manganese. Thereby, the objective of the present study was evaluated the genotoxic damage in juvenile of Geophagus brasiliensis (Quoy e Gaimard, 1824) exposed to Doce river water before (DRWBA - Doce River water before acident) and after (DRWAA - Doce River water after acident) the influx of tailings from the Germano and Santarém Dam disasters in Mariana, MG, Brazil. For this, 24 individuals of the species G. brasiliensis (obtained on IFES/ALEGRE fish culture) were submitted to a bioassay with three treatments and eight replicates. The treatments were: 1) Control water (water from the urban water supply system, filtered with a 0.45 µm membrane), 2) DRBA and 3) DRAA. After 96 h, these fishes were anesthetized to remove blood for evaluation of genotoxic damage (micronucleus and comet). For the bioassay, a total of 80 L of The Doce River water were collected before the influx of tailings and after the influx and then submitted to metal quantification analysis. Fish exposed to DRWBA and DRWAA treatments showed a significant increase in both the number of erythrocyte micronuclei and the DNA damage index in relation to the control fish; however, they did not present any differences between the two treatments. The results demonstrate that the DRWBA treatment was already genotoxic for the fish, mainly due to dissolved Cu concentrations in the water. The DRWAA treatment probably presented genotoxicity due to the increase in the dissolved fraction and synergistic effects of several metals found in the tailings of the Mariana accident.

Resumo A bacia do Rio Doce sofreu o maior acidente ambiental com o influxo de rejeitos de Fundão e Santarém, pertencentes à empresa de mineração Samarco, devido ao desastre em Mariana. Um derramamento entre 50 e 60 milhões de m3 de rejeitos foi estimado pela empresa. De acordo com a Samarco, o rejeito despejado era composto principalmente de argila, silte e alguns metais pesados como ferro, cobre e manganês. Com isso, o presente estudo teve como objetivo avaliar os danos genotóxicos em juvenis de Geophagus brasilienses expostos a água do rio Doce antes (DRWAA - água do Rio Doce antes do acidente) e depois (DRWBA- água do Rio Doce depois do acidente) da chegada dos rejeitos do rompimento das barragens de Germano e Santarém em Mariana, MG, Brasil. Para isso, 24 indivíduos da espécie G. brasilienses (obtidos na piscicultura do IFES/ALEGRE) foram submetidos a um bioensaio com três tratamentos e oito réplicas. Os tratamentos eram: 1) Controle (com água do abastecimento urbano, filtrada com filtro analítico de 0,45 µm); 2) DRWBA e 3) DRWAA. Após um período de 96 h, esses peixes foram anestesiados para retirada de sangue para avaliação dos danos genotóxicos (micronúcleo e cometa). Para a realização do bioensaio, um total de 80 L de água do Rio Doce foram coletados antes da chegada dos rejeitos e outros 80 L foram coletados depois da chegada dos rejeitos e ambas foram submetidas a análises de quantificação de metal. Os peixes expostos ao DRWBA e ao DRWAA apresentaram um aumento significativo na quantidade de micronúcleos eritrocitários e no índice de danos do DNA em relação aos peixes controle, no entanto não apresentaram diferenças entre si. Os resultados obtidos demonstram que a DRWBA já era genotóxica para os peixes, principalmente, em função das concentrações de Cu dissolvido na água. A DRWAA apresentou genotixicidade, provavelmente, em função do aumento da fração dissolvida e do efeito sinérgico de diversos metais presentes nos rejeitos do acidente de Mariana.

Genotoxicity effects on Geophagus brasiliensis fish exposed to Doce River water after the environmental disaster in the city of Mariana, MG, Brazil.

The Doce River basin has suffered the largest environmental accident ever occurred in Brazil with the influx of tailings from Fundão and Santarém, belonging to Samarco mining company, due to the disaster in Mariana. A spill between 50 and 60 million m3 of tailings was estimated by the company. According to Samarco, the wastewater was composed mainly of clay, silt and heavy metals like iron, copper and manganese. Thereby, the objective of the present study was evaluated the genotoxic damage in juvenile of Geophagus brasiliensis (Quoy e Gaimard, 1824) exposed to Doce river water before (DRWBA - Doce River water before acident) and after (DRWAA - Doce River water after acident) the influx of tailings from the Germano and Santarém Dam disasters in Mariana, MG, Brazil. For this, 24 individuals of the species G. brasiliensis (obtained on IFES/ALEGRE fish culture) were submitted to a bioassay with three treatments and eight replicates. The treatments were: 1) Control water (water from the urban water supply system, filtered with a 0.45 µm membrane), 2) DRBA and 3) DRAA. After 96 h, these fishes were anesthetized to remove blood for evaluation of genotoxic damage (micronucleus and comet). For the bioassay, a total of 80 L of The Doce River water were collected before the influx of tailings and after the influx and then submitted to metal quantification analysis. Fish exposed to DRWBA and DRWAA treatments showed a significant increase in both the number of erythrocyte micronuclei and the DNA damage index in relation to the control fish; however, they did not present any differences between the two treatments. The results demonstrate that the DRWBA treatment was already genotoxic for the fish, mainly due to dissolved Cu concentrations in the water. The DRWAA treatment probably presented genotoxicity due to the increase in the dissolved fraction and synergistic effects of several metals found in the tailings of the Mariana accident.

The impact of repeat FNA in non-diagnostic and indeterminate thyroid nodules: A 5-year single-centre experience.

INTRODUCTION: FNA is a well-established method for the preoperative diagnosis of thyroid nodules, but limitations still reside among non-diagnostic and indeterminate samples. The objective of the present study was to assess the impact of repeat FNA in thyroid nodules primarily classified as non-diagnostic and indeterminate, with the evaluation of the diagnostic resolution rate after the reassessment of the nodule. METHODS: We retrospectively collected all cases of thyroid FNA at our institution in the last 5 years that had one or more repeat aspirations of the same nodule, calculating the percentage of samples with change in the diagnostic category. Additional collected data included sex, age and interval between the repeat aspirations. RESULTS: One hundred and seventy-eight specimens from 167 patients (140 female, 27 male) with a median age of 56 years (range 11-90) were included in the study. Among the 86 cases primarily classified as non-diagnostic, 25 (29.1%) remained in the same category after the first reassessment and only 18 (20.9%) after the second repeat aspiration. Among the 40 indeterminate cases, only 10 (25%) retained their status after the second aspiration, with no change after the third assessment. CONCLUSION: Repeat aspiration of non-diagnostic and indeterminate thyroid nodules had a positive impact in both groups, with diagnostic resolution rates of 80% and 75%, respectively. The present study therefore endorses the use of such strategy for the initial follow-up of nodules with no definite diagnosis, especially in low-resource centres with limited access to modern molecular technologies.

Genotoxicity effects on Geophagus brasiliensis fish exposed to Doce River water after the environmental disaster in the city of Mariana, MG, Brazil

Abstract The Doce River basin has suffered the largest environmental accident ever occurred in Brazil with the influx of tailings from Fundão and Santarém, belonging to Samarco mining company, due to the disaster in Mariana. A spill between 50 and 60 million m3 of tailings was estimated by the company. According to Samarco, the wastewater was composed mainly of clay, silt and heavy metals like iron, copper and manganese. Thereby, the objective of the present study was evaluated the genotoxic damage in juvenile of Geophagus brasiliensis (Quoy e Gaimard, 1824) exposed to Doce river water before (DRWBA Doce River water before acident) and after (DRWAA Doce River water after acident) the influx of tailings from the Germano and Santarém Dam disasters in Mariana, MG, Brazil. For this, 24 individuals of the species G. brasiliensis (obtained on IFES/ALEGRE fish culture) were submitted to a bioassay with three treatments and eight replicates. The treatments were: 1) Control water (water from the urban water supply system, filtered with a 0.45 µm membrane), 2) DRBA and 3) DRAA. After 96 h, these fishes were anesthetized to remove blood for evaluation of genotoxic damage (micronucleus and comet). For the bioassay, a total of 80 L of The Doce River water were collected before the influx of tailings and after the influx and then submitted to metal quantification analysis. Fish exposed to DRWBA and DRWAA treatments showed a significant increase in both the number of erythrocyte micronuclei and the DNA damage index in relation to the control fish; however, they did not present any differences between the two treatments. The results demonstrate that the DRWBA treatment was already genotoxic for the fish, mainly due to dissolved Cu concentrations in the water. The DRWAA treatment probably presented genotoxicity due to the increase in the dissolved fraction and synergistic effects of several metals found in the tailings of the Mariana accident.

Resumo A bacia do Rio Doce sofreu o maior acidente ambiental com o influxo de rejeitos de Fundão e Santarém, pertencentes à empresa de mineração Samarco, devido ao desastre em Mariana. Um derramamento entre 50 e 60 milhões de m3 de rejeitos foi estimado pela empresa. De acordo com a Samarco, o rejeito despejado era composto principalmente de argila, silte e alguns metais pesados como ferro, cobre e manganês. Com isso, o presente estudo teve como objetivo avaliar os danos genotóxicos em juvenis de Geophagus brasilienses expostos a água do rio Doce antes (DRWAA água do Rio Doce antes do acidente) e depois (DRWBA- água do Rio Doce depois do acidente) da chegada dos rejeitos do rompimento das barragens de Germano e Santarém em Mariana, MG, Brasil. Para isso, 24 indivíduos da espécie G. brasilienses (obtidos na piscicultura do IFES/ALEGRE) foram submetidos a um bioensaio com três tratamentos e oito réplicas. Os tratamentos eram: 1) Controle (com água do abastecimento urbano, filtrada com filtro analítico de 0,45 µm); 2) DRWBA e 3) DRWAA. Após um período de 96 h, esses peixes foram anestesiados para retirada de sangue para avaliação dos danos genotóxicos (micronúcleo e cometa). Para a realização do bioensaio, um total de 80 L de água do Rio Doce foram coletados antes da chegada dos rejeitos e outros 80 L foram coletados depois da chegada dos rejeitos e ambas foram submetidas a análises de quantificação de metal. Os peixes expostos ao DRWBA e ao DRWAA apresentaram um aumento significativo na quantidade de micronúcleos eritrocitários e no índice de danos do DNA em relação aos peixes controle, no entanto não apresentaram diferenças entre si. Os resultados obtidos demonstram que a DRWBA já era genotóxica para os peixes, principalmente, em função das concentrações de Cu dissolvido na água. A DRWAA apresentou genotixicidade, provavelmente, em função do aumento da fração dissolvida e do efeito sinérgico de diversos metais presentes nos rejeitos do acidente de Mariana.

Trophoblast-macrophage crosstalk on human extravillous under Toxoplasma gondii infection.

INTRODUCTION: The interaction between human extravillous trophoblasts and macrophages has an important role in implantation and placentation. However, any dysfunction in this communication system is associated with pregnancy pitfalls, and a Toxoplasma gondii infection can be a potential problem in this crosstalk. Therefore, the aim of this study was to assess the influence of infected macrophages on cytokine production and the incidence of apoptosis in T. gondii-infected extravillous trophoblast cells. METHODS: HTR-8/SVneo cells were treated with supernatant from macrophages infected or not by T. gondii (conditioned medium) in order to analyze apoptosis and cytokine production in comparison to uninfected control conditions. RESULTS: The IL-6 secretion by HTR-8/SVneo cells increased synergistically by treatment with conditioned medium and T. gondii infection. The apoptosis index of HTR-8/SVneo cells was also upregulated by treatment with conditioned medium and infection. In addition, a low expression of Fas/CD95 and a high soluble FasL release were observed during infection, although no significant change was observed in the proliferation of T. gondii. DISCUSSION: The parasite modulates the high apoptosis index in HTR-8/SVneo cells in order to favor its establishment inside its host cells. On the other hand, the conditioned medium from uninfected macrophages restores the apoptosis rates, although the effect of the infection seems to be stronger. In conclusion, our results showed that T. gondii infection in human extravillous trophoblasts is able to modulate the trophoblast-macrophage crosstalk.

New Insights on Coffea miRNAs: Features and Evolutionary Conservation.

Small RNAs influence the gene expression at the post-transcriptional level by guiding messenger RNA (mRNA) cleavage, translational repression, and chromatin modifications. In addition to model plants, the microRNAs (miRNAs) have been identified in different crop species. In this work, we developed a specific pipeline to search for coffee miRNA homologs on expressed sequence tags (ESTs) and genome survey sequences (GSS) databases. As a result, 36 microRNAs were identified and a total of 616 and 362 potential targets for Coffea arabica and Coffea canephora, respectively. The evolutionary analyses of these molecules were performed by comparing the primary and secondary structures of precursors and mature miRNAs with their orthologs. Moreover, using a stem-loop RT-PCR assay, we evaluated the accumulation of mature miRNAs in genomes with different ploidy levels, detecting an increase in the miRNAs accumulation according to the ploidy raising. Finally, a 5' RACE (Rapid Amplification of cDNA Ends) assay was performed to verify the regulation of auxin responsive factor 8 (ARF8) by MIR167 in coffee plants. The great variety of target genes indicates the functional plasticity of these molecules and reinforces the importance of understanding the RNAi-dependent regulatory mechanisms. Our results expand the study of miRNAs and their target genes in this crop, providing new challenges to understand the biology of these species.