Reproductive biology of Donax striatus (Linnaeus, 1758) (Mollusca: Bivalvia) on Gado Bravo Beach, municipality of Tibau do Norte, state of Rio Grande do Norte, Brazil.

Aspects of the reproductive biology of Donax striatus were studied from individuals collected from Gado Bravo Beach in the municipality of Tibau do Norte, state of Rio Grande do Norte, Brazil. Donax striatus is a dioic species without external (on the shell) or internal (gonads) macroscopic dimorphism. Thus, a microscopic examination of the reproductive cells is necessary. For the characterization of the gonadal development stages and determination of the size at first sexual maturity (L50), 30 specimens were selected monthly between February 2021 and January 2022 and submitted to histological processing. The condition index (CI) of each individual was estimated and monthly variations were statistically assessed. The size at first maturity (L50) was estimated to be 14.2 mm in shell length. To foster conservation of the species, catches of individuals larger than 14.2 mm is recommended. The lowest condition indices were found in the dry season, with a greater occurrence of organisms in the elimination stage and exhibiting gonad tissue reorganization. Higher indices were found in the rainy season, with the presence of mature individuals. The continuous nature of gametogenesis in Donax stritatus reflects the influence of rainfall in the region. Males and females have peak gamete elimination with pauses during the year, but with the presence of maturing and eliminating individuals throughout the year. As shellfish gathering targeting Donax striatus is excessive on Gado Bravo Beach in the state of Rio Grande do Norte, it is hoped that the results of the present study can contribute to the establishment of management measures for the activity and conservation strategies for the species.

Case report: Equine metacarpophalangeal joint partial and full thickness defects treated with allogenic equine synovial membrane mesenchymal stem/stromal cell combined with umbilical cord mesenchymal stem/stromal cell conditioned medium.

Here, we describe a case of a 5-year-old show-jumping stallion presented with severe lameness, swelling, and pain on palpation of the left metacarpophalangeal joint (MCj). Diagnostic imaging revealed full and partial-thickness articular defects over the lateral condyle of the third metacarpus (MC3) and the dorsolateral aspect of the first phalanx (P1). After the lesion's arthroscopic curettage, the patient was subjected to an innovative regenerative treatment consisting of two intra-articular injections of equine synovial membrane mesenchymal stem/stromal cells (eSM-MSCs) combined with umbilical cord mesenchymal stem/stromal cells conditioned medium (UC-MSC CM), 15 days apart. A 12-week rehabilitation program was accomplished, and lameness, pain, and joint effusion were remarkably reduced; however, magnetic resonance imaging (MRI) and computed tomography (CT) scan presented incomplete healing of the MC3's lesion, prompting a second round of treatment. Subsequently, the horse achieved clinical soundness and returned to a higher level of athletic performance, and imaging exams revealed the absence of lesions at P1, fulfillment of the osteochondral lesion, and cartilage-like tissue formation at MC3's lesion site. The positive outcomes suggest the effectiveness of this combination for treating full and partial cartilage defects in horses. Multipotent mesenchymal stem/stromal cells (MSCs) and their bioactive factors compose a novel therapeutic approach for tissue regeneration and organ function restoration with anti-inflammatory and pro-regenerative impact through paracrine mechanisms.

Biochemical analysis of three red grapevine varieties during three phenological periods grown under Mediterranean climate conditions.

In Mediterranean regions, severe summers are becoming more common, leading to restrictions to vine productivity and yield quality, requiring sustainable practices to support this sector. We assessed the behaviour of three red grapevine varieties from the Douro Region to examine their tolerance to summer climate stress from the perspective that the less common varieties may have potential for increased use in a climate change scenario. Leaf and fruit biochemical profile, antioxidant activity and fruit colorimetric parameters were assessed at different phenological stages in Aragonez (AR), Tinto Cão (TC) and Touriga Nacional (TN) grape varieties. All three varieties exhibit significant variability in phenological timing, influenced by genetic and environmental factors. Photosynthetic pigment strategies differed among varieties. Chlorophyll content in AR was high to cope with high radiation, while TN displaying a balanced approach, and TC had lower pigment levels, with higher levels of phenolics, antioxidants, and soluble sugars, particularly during stress. The variations in berry biochemical profile highlight the distinct characteristics of the varieties. TC and TN show potential for coping with climate change, having elevated total acidity, while AR has larger and heavier berries with distinct coloration. These findings reinforce the need to study the behaviour of different varieties in each Terroir, to understand their diverse strategies to deal with summer climate stress. This will help in selecting the most suitable variety for these conditions under vineyard management in the Douro Region.

Early viscoelastometric guided fibrinogen replacement combined with escalation of clinical care reduces progression in postpartum haemorrhage: a comparison of outcomes from two prospective observational studies.

BACKGROUND: Viscoelastometric haemostatic assays (VHA) give rapid information on coagulation status, allowing individualised resuscitation. METHODS: This paper compares outcomes from two observational studies of postpartum haemorrhage (PPH) in the same institution, before and after practice changed from fixed ratio empirical transfusion of coagulation products with laboratory coagulation testing to VHA-guided fibrinogen replacement incorporated into an enhanced PPH care bundle. In both studies, all blood samples were taken near 1000 mL qualitative blood loss (QBL). In Study One, QBL started once PPH was identified, and resuscitation with coagulation blood products was empirical or based on laboratory tests of coagulation. In Study Two, QBL started at delivery and VHA was used to guide fibrinogen replacement if FIBTEM A5 was <12 mm (Claus fibrinogen ≤2 g/L) or to withhold coagulation products if FIBTEM A5 was >12 mm. RESULTS: Improved PPH outcomes were observed in Study Two, with rates of measured blood loss ≥2500 mL, ≥4 units red blood cell (RBC) transfusion, fresh frozen plasma transfusion and ≥8 units of any blood product transfusion all reduced (P < 0.01). Clinically significant improvements occurred in women with fibrinogen ≤2 g/L at study entry, where the proportion of women who received ≥4 units RBC transfusion fell from 67% in Study One to 0% in Study Two (P = 0.0007). CONCLUSIONS: These results suggest that use of VHA as part of an early bundle of PPH care targeting fibrinogen ≤2 g/L with fibrinogen concentrate reduces PPH progression. The greatest benefit was seen when fibrinogen levels were ≤2 g/L at first testing.

Lateral incisional hernia. EVEREG registry analysis.

BACKGROUND: The aim of this study is to analyze the data collected on lateral incisional hernias (LIH) in the National Incisional Hernia Registry (EVEREG) and assess current practices and outcomes in LIH repair. METHODS: A retrospective cohort study was conducted using LIH data recorded over 10 years (2012-2022). Comorbidities, hernia characteristics, short-term complications, and recurrences were studied, along with their association with the type of approach used, either open or laparoscopic, in elective surgery. RESULTS: 1742 LIH cases were studied. According to the EHS classification, these included L1 409 (23.5%), L2 388 (22.3%), L3 565 (32.4%), L4 150 (8.6%) and combined 230 (13.2%). An open approach was performed in 1528 (87.7%) cases and laparoscopic in 214 (12.3%). The median age was 66 ± 12.45 years, with a majority of males, 934 (53.6%). The median body mass index was 29 ± 5.18 kg/m2. The most observed comorbidity was arterial hypertension (957 patients, 55%). A specialist was present in 638 interventions (56%). The 24-month follow-up was 17.9%, and recurrence in those cases was 27.2%, with a higher incidence when there was no specialist present during the intervention, onlay mesh position, and larger defect size. CONCLUSIONS: Surgery for LIH is common, although laparoscopic approach remains infrequent. Furthermore, it is associated with a high percentage of recurrences that increases when there is no specialist in abdominal wall surgery present.

Efficient computational modeling of electronic stopping power of organic polymers for proton therapy optimization.

This comprehensive study delves into the intricate interplay between protons and organic polymers, offering insights into proton therapy in cancer treatment. Focusing on the influence of the spatial electron density distribution on stopping power estimates, we employed real-time time-dependent density functional theory coupled with the Penn method. Surprisingly, the assumption of electron density homogeneity in polymers is fundamentally flawed, resulting in an overestimation of stopping power values at energies below 2 MeV. Moreover, the Bragg rule application in specific compounds exhibited significant deviations from experimental data around the stopping maximum, challenging established norms.

Prelacrimal window approach to the maxillary sinus: a systematic review and meta-analysis of the literature.

BACKGROUND: The prelacrimal window approach (PLWA) is a minimally invasive surgical technique that has been proposed as an alternative to the traditional approaches to access the maxillary sinus. METHODOLOGY: A systematic review with meta-analysis was performed following PRISMA guidelines and identified 368 articles for initial review of which 14 (610 participants) met the criteria for meta-analysis. Four databases, including PubMed, Google Scholar, Web of Science and Scopus, were searched to identify relevant articles. Two independent reviewers conducted the eligibility assessment for the included studies. Methodology quality and risk of bias were evaluated by New Castle Ottawa scale. The outcomes assessed were recurrence of the pathology, postoperative morbidity including epiphora, dry nose, facial, gingival numbness, epistaxis or local infection. RESULTS: The present data suggest a significant reduction in the recurrence rate of maxillary sinus pathology following PLWA when compared to conventional surgery (endoscopic medial maxillectomy, endoscopic sinus surgery and the Caldwell-Luc operation). The rates of epiphora, facial or gingival numbness, epistaxis or infection requiring intervention, were not significantly different between the procedures. CONCLUSIONS: Maxillary sinus pathology can be effectively treated using the PLWA technique, as it has been shown to result in a lower recurrence rate compared to conventional surgeries.

Hypoxia-induced immortalization of primary cells depends on Tfcp2L1 expression.

Cellular senescence is a stress response mechanism that induces proliferative arrest. Hypoxia can bypass senescence and extend the lifespan of primary cells, mainly by decreasing oxidative damage. However, how hypoxia promotes these effects prior to malignant transformation is unknown. Here we observed that the lifespan of mouse embryonic fibroblasts (MEFs) is increased when they are cultured in hypoxia by reducing the expression of p16INK4a, p15INK4b and p21Cip1. We found that proliferating MEFs in hypoxia overexpress Tfcp2l1, which is a main regulator of pluripotency and self-renewal in embryonic stem cells, as well as stemness genes including Oct3/4, Sox2 and Nanog. Tfcp2l1 expression is lost during culture in normoxia, and its expression in hypoxia is regulated by Hif1α. Consistently, its overexpression in hypoxic levels increases the lifespan of MEFs and promotes the overexpression of stemness genes. ATAC-seq and Chip-seq experiments showed that Tfcp2l1 regulates genes that control proliferation and stemness such as Sox2, Sox9, Jarid2 and Ezh2. Additionally, Tfcp2l1 can replicate the hypoxic effect of increasing cellular reprogramming. Altogether, our data suggest that the activation of Tfcp2l1 by hypoxia contributes to immortalization prior to malignant transformation, facilitating tumorigenesis and dedifferentiation by regulating Sox2, Sox9, and Jarid2.

Guarana propagation strategies: a review.

Guarana [Paullinia cupana var. sorbilis (Mart.) Ducke] is a species of great economic and social important in Brazil, as it is the only commercial guarana producer in the world. The vegetative propagation method indicated for the culture is stem cuttings, which aims at productivity, tolerance, and uniformity of clonal cultivars, because reproduction by seeds has slow germination and high genetic variability, which in traditional varieties is an undesirable factor. Genetic factors can interfere with the rooting capacity of the crop. Studies seek alternatives that can improve this condition and enhance the production system. Use of growth regulators, microorganisms that promote plant growth, variation of substrates and fertilization, have been strategies used. Preliminary tests on the rate of stem rooting and seed germination with the use of exogenous phytohormone did not demonstrate in relation to the non-application of these inducers. The use of rhizobacteria, which presents itself as a promising activity in many cultures, has not yet been demonstrated in the culture of guarana. On the other hand, the influence of different substrates on rooting has already shown consistent results as a function of rooting rate. Fertilizing the mother plants as recommended by the production system for the crop has proven to be an efficient procedure. There are still few studies aimed at improving the spread of guarana, demonstrating that new protocols need to be explored, or that the protocols already used are reviewed from another perspective.

Early detection of interstitial lung disease in rheumatic diseases: A joint statement from the Portuguese Pulmonology Society, the Portuguese Rheumatology Society, and the Portuguese Radiology and Nuclear Medicine Society.

INTRODUCTION: Interstitial lung disease (ILD) contributes significantly to morbidity and mortality in connective tissue disease (CTD). Early detection and accurate diagnosis are essential for informing treatment decisions and prognosis in this setting. Clear guidance on CTD-ILD screening, however, is lacking. OBJECTIVE: To establish recommendations for CTD-ILD screening based on the current evidence. METHOD: Following an extensive literature research and evaluation of articles selected for their recency and relevance to the characterization, screening, and management of CTD-ILD, an expert panel formed by six pulmonologists from the Portuguese Society of Pulmonology, six rheumatologists from the Portuguese Society of Rheumatology, and six radiologists from the Portuguese Society of Radiology and Nuclear Medicine participated in a multidisciplinary discussion to produce a joint statement on screening recommendations for ILD in CTD. RESULTS: The expert panel achieved consensus on when and how to screen for ILD in patients with systemic sclerosis, rheumatoid arthritis, mixed connective tissue disease, Sjögren syndrome, idiopathic inflammatory myopathies and systemic lupus erythematous. CONCLUSIONS: Despite the lack of data on screening for CTD-ILD, an expert panel of pulmonologists, rheumatologists and radiologists agreed on a series of screening recommendations to support decision-making and enable early diagnosis of ILD to ultimately improve outcomes and prognosis in patients with CTD.

A semi-automated microscopic image analysis method for scoring Ki-67 nuclear immunostaining.

Nuclear proliferation marker MIB-1 (Ki-67) immunohistochemistry (IHC) is used to examine tumor cell proliferation. However, the diagnostic or prognostic value of the Ki-67 nuclear staining intensity and location, defined as nuclear gradient (NG), has not been assessed. This study examined the potential association between Ki-67 NG and cell cycle phases and its effect on the prognosis of pulmonary typical carcinoid (PTC) tumors. We propose a method for classifying the NG of Ki-67 during the cell cycle and compare the results between PTC, pulmonary adenocarcinoma (PAD), and breast ductal carcinoma (BDC). A literature review and objective analysis of IHC-stained paraffin sections were used to determine the Ki-67 labeling index and composed a stratification of the NG into NG1, NG2, and NG3/4 categories. A semi-automated image analysis protocol was established to determine the Ki-67 NG in PTC, PAD, and BDC. High intraobserver consistency and moderate interobserver agreement were achieved in the determination of Ki-67 NG in tumor specimens. NG1 and NG2 were lower in PTC than in PAD and BDC. Cox multivariate analysis of PTC after adjusting for age and number of metastatic lymph nodes showed that Ki-67 NG1 and NG2 significantly predicted clinical outcomes. The semi-automated method for quantification of Ki-67 nuclear immunostaining proposed in this study could become a valuable diagnostic and prognostic tool in PTC.

Association between rainfall and Escherichia coli in live bivalve molluscs harvested in Sardinia, Italy.

Rainfall is generally accepted as one of the most important factors associated with an increased level of E. coli in bivalve molluscs. Performing microbiological risk assessment is relevant to official control authorities to determine the sanitary status of harvesting areas and, therefore, develop monitoring strategies and identify management practices that could be used to improve the quality and safety of the final product. The present study aimed to investigate the impact of rainfall on the content of E. coli in bivalve molluscs farmed in Sardinia (Italy). Enumeration of E. coli was performed according to the Most Probable Number (MPN) method (ISO 16649-3) on 1,920 bivalve samples collected from 7 regional counties between 2018 and 2020. Bivalve molluscs samples included 955 mussels (Mytilus galloprovincialis), 500 oysters (Crassostrea gigas), 325 clams (Ruditapes decussatus), 94 warty venus (Venus verrucosa), and 46 lagoon cockles (Cerastoderma glaucum). Rainfall data were obtained by the Department of Meteorology of the ARPA Sardegna. For each sampling site, GPS coordinates were used to identify gauge stations within catchment areas. Cumulative rain (mm) was recorded 1, 3, 5, 7, and 15 days before sampling, among which the 7-day cumulative rain was the strongest predictor of E. coli counts. Several thresholds of 7-day cumulative rain (from <10 mm up to >300 mm) before sampling were used to estimate the chances of a non-compliant sample (E. coli levels above the limit for sanitary class A; 230 MPN/100 g). The 7-day cumulative rain was positively associated with the chances of non-compliance. When the 7-day cumulative rain before sampling was >300 mm, 80.5 % of the samples were non-compliant, and the odds of a non-compliant sample were 23.6 times higher, as compared to samples harvested when the 7-day cumulative rainfall was <10 mm. Precipitation data could be a useful tool for interpreting anomalous results from official control authorities and reduce the costs that originate from closure of production areas.

Medium-term evaluation of the bond strength and dentin penetration of self-adhesive resin cements to root dentin.

BACKGROUND: Medium-term evaluate the bond strength and tag formation in root dentin using self-adhesive cements Maxcem Elite (MCE), Relyx U200 (RU2), SeT PP (SPP) and Megalink (MGL) compared to conventional cement Relyx ARC (ARC) for glass fibre post cementation. MATERIALS AND METHODS: One hundred bovine incisors roots were endodontically treated and divided into 5 groups (n = 20) according to the cementation system: MCE, RU2, SPP, MGL and ARC. All cementation systems were evaluated at 24 h (1) and 12 months (2) after post cementation. Specimens from the cervical, middle and apical thirds of the root were submitted to push-out bond strength test and confocal laser microscopy to verify the dentin penetration. The adhesive failure mode was classified as adhesive, cohesive and mixed. Data were analysed by one-way ANOVA and Tukey tests (α = 5%). RESULTS: ARC-1 and ARC-12 showed the highest bond strength (P < 0.05). ARC showed the greatest tag extension, regardless of the third (P < 0.05). The most frequent failure mode in the 24-h analysis was cohesive, regardless of the cement used. For the 12-month analysis, mixed failure was the most frequent. CONCLUSION: Conventional cement (ARC) showed the highest bond strength and tag extension, regardless of the evaluation period. In all cementation systems, the bond strength decreased after 12 months. © 2023 Australian Dental Association.

Mechanical osteoarthritis of the hip in a one medicine concept: a narrative review.

Human and veterinary medicine have historically presented many medical areas of potential synergy and convergence. Mechanical osteoarthritis (MOA) is characterized by a gradual complex imbalance between cartilage production, loss, and derangement. Any joint instability that results in an abnormal overload of the joint surface can trigger MOA. As MOA has a prevailing mechanical aetiology, treatment effectiveness can only be accomplished if altered joint mechanics and mechanosensitive pathways are normalized and restored. Otherwise, the inflammatory cascade of osteoarthritis will be initiated, and the changes may become irreversible. The management of the disease using non-steroidal anti-inflammatory drugs, analgesics, physical therapy, diet changes, or nutraceuticals is conservative and less effective. MOA is a determinant factor for the development of hip dysplasia in both humans and dogs. Hip dysplasia is a hereditary disease with a high incidence and, therefore, of great clinical importance due to the associated discomfort and significant functional limitations. Furthermore, on account of analogous human and canine hip dysplasia disease and under the One Medicine concept, unifying veterinary and human research could improve the well-being and health of both species, increasing the acknowledgement of shared diseases. Great success has been accomplished in humans regarding preventive conservative management of hip dysplasia and following One Medicine concept, similar measures would benefit dogs. Moreover, animal models have long been used to better understand the different diseases' mechanisms. Current research in animal models was addressed and the role of rabbit models in pathophysiologic studies and of the dog as a spontaneous animal model were highlighted, denoting the inexistence of rabbit functional models to investigate therapeutic approaches in hip MOA.

Aerobic exercise prevents cardiomyocyte damage caused by oxidative stress in early cardiovascular disease by increasing vascularity while L-arginine supplementation prevents it by increasing activation of the enzyme nitric oxide synthase.

L-Arginine and chronic exercise reduce oxidative stress. However, it is unclear how they affect cardiomyocytes during cardiovascular disease (CVD) development. The aim of this research was to investigate the possible effects of L-arginine supplementation and aerobic training on systemic oxidative stress and their consequences on cardiomyocytes during cardiometabolic disease onset caused by excess fructose. Wistar rats were allocated into four groups: control (C), fructose (F, 10% fructose in water), fructose training (FT; moderate running, 50-70% of the maximal velocity), and fructose arginine (FA; 880 mg/kg/day). Fructose was given for two weeks and fructose plus treatments for the subsequent eight weeks. Body composition, blood glucose, insulin, lipid profile, lipid peroxidation, nitrite, metalloproteinase-2 (MMP-2) activity, left ventricle histological changes, microRNA-126, -195, and -146, eNOS, p-eNOS, and TNF-α expressions were analyzed. Higher abdominal fat mass, triacylglycerol level, and insulin level were observed in the F group, and both treatments reversed these alterations. Myocardial vascularization was impaired in fructose-fed groups, except in FT. Cardiomyocyte hypertrophy was observed in all fructose-fed groups. TNF-α levels were higher in fructose-fed groups than in the C group, and p-eNOS levels were higher in the FA than in the C and F groups. Lipid peroxidation was higher in the F group than in the FT and C groups. During CVD onset, moderate aerobic exercise reduced lipid peroxidation, and both training and L-arginine prevented metabolic changes caused by excessive fructose. Myocardial vascularization was impaired by fructose, and cardiomyocyte hypertrophy appeared to be influenced by pro-inflammatory and oxidative environments.

Thoracoscopic pneumonectomy in a 4 year-old-child with destroyed lung following viral pneumonia. / Tiroidectomía transoral endoscópica con abordaje vestibular (TOETVA): una nueva opción para el tratamiento de pacientes pediátricos.

Pediatric pneumonectomies are exceptional nowadays, being reserved for cases with destroyed lungs with frequent exacerbations and reinfections and only two cases of thoracoscopic pneumonectomy have been previously published. We present the case of a 4-year-old patient with no relevant history who developed complete atelectasis of the left lung (LL) after influenza A pneumonia, followed by secondary recurrent infections. A year later a diagnostic bronchoscopy without alterations was performed. A complete loss of volume and hypoperfusion of the LL (right lung perfusion 95%, LL perfusion: 5%) with bronchiectasis and hyperinsufflation and herniation of the right lung into the left hemithorax was observed in a pulmonary perfusion SPECT-CT. After unsuccessful conservative management and recurrent infections a pneumonectomy was indicated. The pneumonectomy was performed through a five-port thoracoscopy. The dissection of the hilum was made using hook electrocautery and sealing device. The left main bronchus was sectioned with an endostapler. There were no intraoperative complications. An endothoracic drain was removed the first postoperative day. The patient was discharged on the fourth postoperative day. The patient has not presented any complications 10 months after surgery. Although pneumonectomy is an exceptional surgery in children, it can be performed by minimally invasive surgery with success and safety in centers with extensive experience in pediatric thoracoscopic surgery.

Hoy en día, las neumonectomías pediátricas son algo excepcional. El procedimiento se reserva para aquellos casos en los que los pulmones están destruidos y presentan exacerbaciones y reinfecciones frecuentes, con tan solo dos casos de neumonectomía toracoscópica publicados hasta la fecha. Presentamos el caso de un paciente de 4 años sin antecedentes de interés que desarrolló atelectasia completa del pulmón izquierdo (PI) tras neumonía por gripe A, seguido de infecciones secundarias recurrentes. Un año después, se le practicó broncoscopia diagnóstica, sin que esta mostrara alteraciones significativas. Tras realizársele un SPECT-CT de perfusión pulmonar, se evidenció pérdida completa de volumen e hipoperfusión del PI (perfusión del pulmón derecho: 95%; perfusión del pulmón izquierdo: 5%), con bronquiectasia e hiperinsuflación y herniación del pulmón derecho hacia el hemitórax izquierdo. Tras fracasar el manejo conservador y registrarse infecciones recurrentes, se estableció la indicación de neumonectomía. La neumonectomía se llevó a cabo mediante toracoscopia por cinco puertos. La disección del hilio se realizó mediante gancho de electrocoagulación y dispositivo de sellado. El bronquio principal izquierdo se seccionó con endograpadora. No se registraron complicaciones intraoperatorias. El drenaje endotorácico se retiró al día siguiente de la intervención, mientras que el paciente fue dado de alta a los cuatro días, sin que haya presentado complicaciones transcurridos 10 meses desde la cirugía. Aunque la neumonectomía es una intervención excepcional en niños, puede llevarse a cabo de manera exitosa y segura por cirugía mínimamente invasiva en centros con amplia experiencia en cirugía toracoscópica pediátrica.

Evaluation of a battery of biotests to improve waste ecotoxicity assessment (HP 14), using incineration bottom ash as a case study.

The assessment of waste ecotoxicity (hazardous property HP14 in the European Union) is fundamental for proper waste classification and safe application/disposal. Biotests are relevant for evaluating waste complex matrices, but their efficiency is crucial to encourage their adoption at the industrial level. This work aims at evaluating possibilities of improving the efficiency of a biotest battery previously suggested in the literature, regarding test selection, duration, and/or laboratory resources optimization. Fresh incineration bottom ash (IBA) was the case study. The test battery analysed included standard aquatic (bacteria, microalgae, macrophytes, daphnids, rotifers, fairy shrimp) and terrestrial (bacteria, plants, earthworms, collembolans) organisms. The assessment followed an Extended Limit Test design (three dilutions of eluate or solid IBA) and the Lowest Ineffective Dilution (LID-approach) for ecotoxicity classification. The results emphasize the importance of testing different species. It was also evidenced that tests with daphnids and earthworms may be shortened to 24 h; the miniaturization of tests is suitable as e.g. differential sensitivity of microalgae and macrophytes was captured with low variability; alternative testing kits can be used when methodological difficulties are found. Microalgae were more sensitive than macrophytes. Similar results were found for the Thamnotoxkit and daphnids test for eluates with natural pH, so the former may be used as an alternative. B. rapa was the most sensitive organism, suggesting that it may be tested as the only terrestrial plant species and that minimum test duration is appropriate. F. candida does not appear to add information to the battery. The differences in sensitivity of A. fischeri and E. fetida compared to the remaining species were not significant enough to exclude them from the battery. Thus, this work suggests a biotest battery to test IBA comprising aquatic tests - Aliivibrio fischeri, Raphidocelis subcapitata (miniaturised test), and Daphnia magna (24 h when clear deleterious effects are observed) or Thamnocephalus platyurus (toxkit) - and terrestrial tests - Arthrobacter globiformis, Brassica rapa (14 d), and Eisenia fetida (24 h). Testing waste with natural pH is also recommended. The Extended Limit Test design considering the LID-approach seems useful in waste testing, particularly for the industry, involving low effort, test material requirements, and few laboratory resources. The LID-approach allowed for differentiating ecotoxic from non-ecotoxic effects and captured different sensitivities between species. Ecotoxicological assessment of other waste may benefit from these recommendations, but caution should be taken given the properties of each waste type.

Lower low density lipoprotein cholesterol associates to higher mortality in non-diabetic heart failure patients.

Background: In patients with established heart failure (HF) low total cholesterol levels associate with worse prognosis. Evidence concerning the impact of Low-density lipoprotein cholesterol (LDL-c) in HF is scarce. We aimed to evaluate the prognostic impact of LDL-c in patients with HF, both with and without diabetes mellitus (DM). Methods: We retrospectively analyzed outpatients with chronic HF with systolic dysfunction followed in our HF clinic from January/2012 to May/2018. LDL-c was calculated using the Friedewald's formula. Patients without a complete lipid profile were excluded. The endpoint under analysis was all-cause mortality. Patients were followed until January/2021. A Cox-regression analysis was used to study the prognostic impact of LDL-c. The LDL-c cut-off used was 100 mg/dL (mean value). Analysis was stratified according to the coexistence of DM. Multivariate models were built adjusting for age, sex, coronary artery disease, atherosclerotic non-coronary artery disease, arterial hypertension, smoking status, statin use, severity of systolic dysfunction, creatinine clearance and evidence-based therapy. Results: We studied 522 chronic HF patients, mean age was 70 years, 66.5% males. Severe systolic dysfunction was present in 42.7%, 30.5% had coronary heart disease, 60.5% had arterial hypertension, 41.6% had DM. A total of 92.0% were treated with beta blocker, 87.5% with an ACEi/ARB and 29.1% with a MRA. During a median follow-up of 53 (interquartile range 33-73) months, 235 (45%) patients died. Patients with LDL-c ≤100 mg/dL presented increased multivariate-adjusted risk of all-cause mortality: HR = 1.58 (95% CI: 1.08-2.30), p = 0.02. When patients were stratified according to DM, LDL-c ≤100 mg/dL was independently associated with increased death risk - HR = 1.55 (95% CI:1.05-2.30), p = 0.03 in patients without DM; in patients with DM no association was detected - multivariate-adjusted HR = 1.18 (95% CI: 0.77-1.80), p = 0.44. Conclusion: Non-DM HF patients with LDL-c>100 mg/dL have a 35% reduction in the mortality risk when compared with those with lower values. The "cholesterol paradox" in HF also applies to LDL-c in non-DM patients.

A pro-BMP function exerted by Rhodnius prolixus short gastrulation reveals great diversity in the role of BMP modulators during embryonic patterning.

Dorsal-ventral (DV) patterning is regulated by the bone morphogenetic pathway (BMP) in Bilateria. In insect DV patterning, the Toll pathway also plays a role, in addition to BMPs. Variations in the relative importance of each pathway for DV patterning have been reported using single species of coleopteran, hymenopteran, hemipteran and orthopteran insects. To investigate if the molecular control of DV patterning is conserved inside an insect order, the emergent model hemiptera species Rhodnius prolixus was studied. We found that R. prolixus BMP pathway controls the entire DV axis, with a broader effect respective to Toll, as shown for the hemiptera Oncopeltus fasciatus. Different from O. fasciatus, the unique R. prolixus short gastrulation (sog) and the twisted gastrulation (tsg) orthologues do not antagonize, but rather favour embryonic BMP signalling. Our results reinforce the hypothesis that hemiptera rely preferentially on BMPs for DV patterning but that, surprisingly, in R. prolixus Sog and Tsg proteins exert only a positive role to establish a dorsal-to-ventral BMP gradient. Since sog has been reported to be lost from orthopteran and hymenopteran genomes, our results indicate that Sog's role to modify BMP activity varies greatly in different insect species.