Exploring Immunohistochemistry in Fish: Assessment of Antibody Reactivity by Western Immunoblotting.

In recent years, research on fish has seen remarkable advancements, especially in aquaculture, ornamental fish industry, and biomedical studies. Immunohistochemistry has become crucial in fish research, aiding in physiological and pathological investigations. However, the use of antibodies originally developed for mammals has raised concerns about their cross-reactivity and specificity in fish. This study systematically evaluated the reactivity of commonly used antibodies for diagnostic purposes, especially in fish pathology, including pan-cytokeratin, vimentin, S-100, glial fibrillary acidic protein, and desmin in the tissue of Sparus aurata, Dicentrarchus labrax, Oncorhynchus mykiss, and Carassius auratus. Western immunoblotting was employed to assess antibody specificity. The results revealed that the pan-cytokeratin and glial fibrillary acidic protein antibodies cross-react with all tested fish species, while S-100 demonstrated specific staining in sea bream, goldfish, and rainbow trout tissues. Conversely, vimentin and desmin antibodies displayed no reactivity. In conclusion, the anti-cytokeratin clone AE1/AE3 and the polyclonal rabbit anti-glial fibrillary acidic protein antibody, which are extensively used in mammals, were validated for fish immunohistochemical studies. Regrettably, D33 anti-desmin and V9 anti-vimentin clones are unsuitable for immunohistochemistry in the tested fish. These findings underscore the need for species-specific antibodies and proper validation for accurate immunohistochemistry analyses in fish research.

Canine Mammary Tumor Histopathological Image Classification via Computer-Aided Pathology: An Available Dataset for Imaging Analysis.

Histopathology, the gold-standard technique in classifying canine mammary tumors (CMTs), is a time-consuming process, affected by high inter-observer variability. Digital (DP) and Computer-aided pathology (CAD) are emergent fields that will improve overall classification accuracy. In this study, the ability of the CAD systems to distinguish benign from malignant CMTs has been explored on a dataset-namely CMTD-of 1056 hematoxylin and eosin JPEG images from 20 benign and 24 malignant CMTs, with three different CAD systems based on the combination of a convolutional neural network (VGG16, Inception v3, EfficientNet), which acts as a feature extractor, and a classifier (support vector machines (SVM) or stochastic gradient boosting (SGB)), placed on top of the neural net. Based on a human breast cancer dataset (i.e., BreakHis) (accuracy from 0.86 to 0.91), our models were applied to the CMT dataset, showing accuracy from 0.63 to 0.85 across all architectures. The EfficientNet framework coupled with SVM resulted in the best performances with an accuracy from 0.82 to 0.85. The encouraging results obtained by the use of DP and CAD systems in CMTs provide an interesting perspective on the integration of artificial intelligence and machine learning technologies in cancer-related research.

Cosmetic Results and Side Effects of Accelerated Partial-Breast Irradiation Versus Whole-Breast Irradiation for Low-Risk Invasive Carcinoma of the Breast: The Randomized Phase III IRMA Trial.

PURPOSE: The results in terms of side effects vary among the published accelerated partial-breast irradiation (APBI) studies. Here, we report the 5-year results for cosmetic outcomes and toxicity of the IRMA trial. METHODS: We ran this randomized phase III trial in 35 centers. Women with stage I-IIA breast cancer treated with breast-conserving surgery, age ≥ 49 years, were randomly assigned 1:1 to receive either whole-breast irradiation (WBI) or external beam radiation therapy APBI (38.5 Gy/10 fraction twice daily). Patients and investigators were not masked to treatment allocation. The primary end point was ipsilateral breast tumor recurrence. We hereby present the analysis of the secondary outcomes, cosmesis, and normal tissue toxicity. All side effects were graded with the Radiation Therapy Oncology Group/European Organisation for Research and Treatment of Cancer Radiation Morbidity Scoring Schema. Analysis was performed with both intention-to-treat and as-treated approaches. RESULTS: Between March 2007 and March 2019, 3,309 patients were randomly assigned to 1,657 WBI and 1,652 APBI; 3,225 patients comprised the intention-to-treat population (1,623 WBI and 1,602 APBI). At a median follow-up of 5.6 (interquartile range, 4.0-8.4) years, adverse cosmesis in the APBI patients was higher than that in the WBI patients at 3 years (12.7% v 9.2%; P = .009) and at 5 years (14% v 9.8%; P = .012). Late soft tissue toxicity (grade ≥ 3: 2.8% APBI v 1% WBI, P < .0001) and late bone toxicity (grade ≥ 3: 1.1% APBI v 0% WBI, P < .0001) were significantly higher in the APBI arm. There were no significant differences in late skin and lung toxicities. CONCLUSION: External beam radiation therapy-APBI with a twice-daily IRMA schedule was associated with increased rates of late moderate soft tissue and bone toxicities, with a slight decrease in patient-reported cosmetic outcomes at 5 years when compared with WBI, although overall toxicity was in an acceptable range.

Ejaculate characteristics over seasons in five species of lancehead pitvipers (Bothrops spp) kept in captivity.

Due to their major medical importance in Latin America, lancehead pitvipers are frequently kept and bred in captivity for venom extraction to the production of antivenom serums. Nevertheless, despite the great contribution given to captive breeding, much of the knowledge of Bothrops' reproductive biology derived from sporadic and insufficient data provided by zoological collections. Thus, we aimed to investigate seasonal changes in gonadosomatic index (GSI) and seminal parameters (e.g., volume, concentration, motility, viability, and acrosome integrity) of five species of lancehead pitvipers from different biomes and phylogenetic groups, maintained in the indoors serpentarium at Butantan Institute (Brazil). Patterns of variation in GSI and semen parameters differed from one species to another, suggesting that captive populations should perhaps be managed distinctly to maximize reproductive success. Furthermore, in none of the studied species did changes in GSI occur concomitantly with seminal variations. GSI remained unaltered year-round for Jararaca (Bothrops jararaca) and Brazilian lancehead (Bothrops moojeni), whereas it peaked in the autumn for Common lancehead (Bothrops atrox), Jararacussu (Bothrops jararacussu), and Whitetail lancehead (Bothrops leucurus). But surprisingly, the scenario was inverted when we estimated the total number of motile spermatozoa per season, as Jararaca and Brazilian lancehead displayed seasonal differences and the other species did not vary throughout the year. Potential ecological and evolutionary factors underlying these differences were also discussed in the present article. Together, these findings can help to better define breeding management strategies for each species in captivity, in addition to optimizing the future use of artificial insemination and semen cryopreservation.

Deep Eutectic Solvents: Alternative Solvents for Biomass-Based Waste Valorization.

Innovative technologies can transform what are now considered "waste streams" into feedstocks for a range of products. Indeed, the use of biomass as a source of biopolymers and chemicals currently has a consolidated economic dimension, with well-developed and regulated markets, in which the evaluation of the manufacturing processes relies on specific criteria such as purity and yield, and respects defined regulatory parameters for the process safety. In this context, ionic liquids and deep eutectic solvents have been proposed as environmentally friendly solvents for applications related to biomass waste valorization. This mini-review draws attention to some recent advancements in the use of a series of new-solvent technologies, with an emphasis on deep eutectic solvents (DESs) as key players in the development of new processes for biomass waste valorization. This work aims to highlight the role and importance of DESs in the following three strategic areas: chitin recovery from biomass and isolation of valuable chemicals and biofuels from biomass waste streams.

Investigations on the Fatigue Behaviour of 3D-Printed Continuous Carbon Fibre-Reinforced Polymer Tension Straps.

The focus of this research is an investigation on the fatigue behaviour of unidirectional 3D-printed continuous carbon fibre-reinforced polymer (CFRP) tension straps with a polyamide matrix (PA12). Conventionally produced tension straps are becoming established components in the mechanical as well as the civil engineering sector, e.g., as rigging systems for sailing boats and cranes and-recently introduced-as network arch bridge hangers. All these structures are subjected to high fatigue loads, and although it is commonly reported that carbon fibre-reinforced polymers show excellent fatigue resistance, there is limited understanding of the behaviour of CFRP loop elements under such loads, especially in combination with fretting at the attachment points. Research on this topic was performed at Empa in the past decade on thermoset CFRP straps, but never before with 3D-printed continuous CFRP straps with a thermoplastic matrix. This paper examines an additive manufacturing and post-consolidation method for producing the straps and presents initial results on their fatigue performance, which show that the fatigue endurance limit of the investigated 3D-printed and post-consolidated CFRP strap design is acceptable, when compared to steel tendons. However, it is still 20% lower than conventionally produced CFRP straps using out-of-autoclave unidirectional carbon fibre prepregs. The reasons for these findings and potential future improvements are discussed.

Economical gold recovery cycle from bio-sensing AuNPs: an application for nanowaste and COVID-19 testing kits.

We report the controlled growth of biologically active compounds: gold nanoparticles (AuNPs) in various shapes, including their green synthesis, characterization, and studies of their applications towards biological, degradation and recycling. Using spectroscopic methods, studies on responsive binding mechanisms of AuNPs with biopolymers herring sperm deoxyribonucleic acid (hsDNA), bovine serum albumin (BSA), dyes degradation study, and exquisitely gold separation studies/recovery from nanowaste, COVID-19 testing kits, and pregnancy testing kits are discussed. The sensing ability of the AuNPs with biopolymers was investigated via various analytical techniques. The rate of degradation of various dyes in the presence and absence of AuNPs was studied by deploying stirring, IR, solar, and UV-Vis methods. AuNPs were found to be the most active cytotoxic agent against human breast cancer cell lines such as MCF-7 and MDAMB-468. Furthermore, an economical process for the recovery of gold traces from nanowaste, COVID-19 detection kits, and pregnancy testing kits was developed using inexpensive and eco-friendly α-cyclodextrin sugar. This method was found to be easy and safest in comparison with the universally accepted cyanidation process. In the future, small gold jewelry makers and related industries would benefit from the proposed gold-recycling process and it might contribute to their socio-economic growth. The methodologies proposed are also beneficial for trace-level forensic investigation.

A Review on Pathological and Diagnostic Aspects of Emerging Viruses-Senecavirus A, Torque teno sus virus and Linda Virus-In Swine.

Swine production represents a significant component in agricultural economies as it occupies over 30% of global meat demand. Infectious diseases could constrain the swine health and productivity of the global swine industry. In particular, emerging swine viral diseases are omnipresent in swine populations, but the limited knowledge of the pathogenesis and the scarce information related to associated lesions restrict the development of data-based control strategies aimed to reduce the potentially great impact on the swine industry. In this paper, we reviewed and summarized the main pathological findings related to emerging viruses, such as Senecavirus A, Torque teno sus virus, and Linda virus, suggesting a call for further multidisciplinary studies aimed to fill this lack of knowledge and better clarify the potential role of those viral diseases in swine pathology.

Reproducibility and Feasibility of Classification and National Guidelines for Histological Diagnosis of Canine Mammary Gland Tumours: A Multi-Institutional Ring Study.

Histological diagnosis of Canine Mammary Tumours (CMTs) provides the basis for proper treatment and follow-up. Nowadays, its accuracy is poorly understood and variable interpretation of histological criteria leads to a lack of standardisation and impossibility to compare studies. This study aimed to quantify the reproducibility of histological diagnosis and grading in CMTs. A blinded ring test on 36 CMTs was performed by 15 veterinary pathologists with different levels of education, after discussion of critical points on the Davis-Thompson Foundation Classification and providing consensus guidelines. Kappa statistics were used to compare the interobserver variability. The overall concordance rate of diagnostic interpretations of WP on identification of hyperplasia-dysplasia/benign/malignant lesions showed a substantial agreement (average k ranging from 0.66 to 0.82, with a k-combined of 0.76). Instead, outcomes on ICD-O-3.2 morphological code /diagnosis of histotype had only a moderate agreement (average k ranging from 0.44 and 0.64, with a k-combined of 0.54). The results demonstrated that standardised classification and consensus guidelines can produce moderate to substantial agreement; however, further efforts are needed to increase this agreement in distinguishing benign versus malignant lesions and in histological grading.

Click gold quantum dots biosynthesis with conjugation of quercetin for adenocarcinoma exertion.

We developed a cost-effective and eco-friendly click biosynthesis of small molecule quercetin-gold quantum dots (QRT-AuQDs) involving quick conjugation using an ultrasonication method at ambient temperature by utilizing QRT and gold ions in the proportion of 0.1 : 1 (molar ratio). A comparatively very short amount of time (60 seconds) was required as compared to conventional procedures. The present biomimetics research relates to the isolation of bioactive QRT by the circularly spread silica gel layer technique (CSSGLT) and characterization (UV-Vis, FTIR, NMR and DSC analysis). Characterization of the synthesized QRT-AuQDs conjugated complex was carried out by UV-Vis, HR-TEM, DLS, zeta potential and X-ray diffraction. The main objective of the present work was to study the comparative anticancer activity of QRT and QRT-AuQDs on human lung cancer HOP-62 and leukemia K-562 cell lines. The results suggested that QRT-AuQDs showed potential for applications in anticancer treatment and were found to be a more cytotoxic agent in comparison to QRT, causing > 50% inhibition of cancer cells at the concentration < 10-7 M. Hence, small molecule conjugated QRT-AuQDs can be used as a promising material for biomedical, bioengineering and anti-infectives applications.

Canine and feline in situ mammary carcinoma: A comparative review.

Carcinoma in situ of the breast is a well-known entity in humans. In veterinary medicine, particularly in canine and feline mammary literature, there is no agreement whether the term in situ should be used to indicate a specific carcinoma histotype or the noninvasive status of a carcinoma of any histotype. Moreover, in the most recent histologic classification of mammary tumors published by the Davis-Thompson Foundation, it is suggested to abandon the term carcinoma in situ given the lack of standardized criteria defining this entity, replacing it with epitheliosis or ductal/lobular hyperplasia with severe atypia. This publication presents a critical review of the term in situ in human and veterinary medicine considering the evolution of the term over the years and its heterogeneous use by different authors, including variations in immunohistochemical markers for classification. This review aims to point out the lack of uniformity in the nomenclature and classification issues in veterinary medicine regarding the use of the term in situ, laying the ground for a process of standardization in future publications.

Total parenteral nutrition is associated with worse hospital outcomes among elderly diabetic patients: a propensity score matched analysis on discharge records.

Background: Recent trials and reviews have raised question about the safety of total parenteral nutrition (TPN), due to the increased rate of TPN related complications. Diabetic patients are vulnerable to hyperglycaemia, and poor studies have investigated hospital out-comes of diabetic patients requiring TPN. The aim of this study was to evaluate the association of in-hospital mortality, prolonged length of stay and transfer to long-term care facilities among diabetic patients with TPN. Methods: The study considered all hospital admissions of diabetic patients over 65 years of age performed between 2006 and 2015 in Abruzzo Region, Italy. To compare the outcomes of TPN and non-TPN patients, a propensity score matching procedure was performed. Results: A total of 140,556 admissions were analyzed. After matching, 1947 patients were included into the analyses: 649 patients with TPN and 1298 controls. TPN was significantly associated to in-hospital mortality (OR=7.15; 95%CI 5.54-9.22), prolonged LOS (OR=2.78; 95%CI 2.28-3.38) and transfer to LTCF (OR=2.16; 95%CI 1.64-2.85). Discussion: TPN is associated with poor outcomes among elderly diabetic patients in the Italian setting. Being aware of the risk factors among diabetic patients with TPN can be used to anticipate the patients' needs during the admission and the immediate post-discharge period.

Predictors of Prolonged Hospitalization and In-Hospital Mortality After Hip Fracture: A Retrospective Study on Discharge Registry.

Background: Hip fracture injury is one of the principal health problems affecting the elderly. Patients reporting hip fractures often show relevant comorbidities leading to prolonged hospital stay, significant complications and higher mortality rates. This study aims to assess the risk factors associated with prolonged hospitalization after hip fracture, in-hospital mortality and transfers to other facilities. Study design: Retrospective cross-sectional study. Methods: The study considered all admissions performed between 2006 and 2015 in Abruzzo region, Italy. Logistic regression analyses were performed to evaluate odds ratios for each risk factor as predictor of in-hospital mortality, length of stay, and transfer to other facilities. Results: Age over 85 (OR=5.38) and cancer (OR=3.62) were identified as the strongest risk predictors for in hospital mortality; diabetes (OR=2.24) and heart failure (OR=1.57) were identified as predictors of prolonged length of stay and age over 85 (OR=1.38) and atrial fibrillation (OR=1.69) were identified as predictors of transfer to other facilities. Conclusions: With the rising incidence of hip fractures, identification of modifiable factors may help to reduce morbidity and mortality.

Fretting Fatigue Performance of Unidirectional, Laminated Carbon Fibre Reinforced Polymer Straps at Elevated Service Temperature.

The fretting fatigue performance of laminated, unidirectional (UD), pin-loaded, carbon fibre-reinforced polymer (CFRP) straps that can be used as bridge hanger cables was investigated at a sustained service temperature of 60 °C. The aim of this paper is to elucidate the influence of the slightly elevated service temperature on the tensile fatigue performance of CFRP straps. First, steady state thermal tests at ambient temperature and at 60 °C are presented, in order to establish the behaviour of the straps at these temperatures. These results indicated that the static tensile performance of the straps is not affected by the increase in temperature. Subsequently, nine upper stress levels (USLs) between 650 and 1400 MPa were chosen in order to establish the S-N curve at 60 °C (frequency 10 Hz; R = 0.1) and a comparison with an existing S-N curve at ambient temperature was made. In general, the straps fatigue limit was slightly decreased by temperature, up to 750 MPa USL, while, for the higher USLs, the straps performed slightly better as compared with the S-N curve at ambient temperature.

A New Dermal Substitute Containing Polyvinyl Alcohol with Silver Nanoparticles and Collagen with Hyaluronic Acid: In Vitro and In Vivo Approaches.

The experimental use of poly (alcohol-vinyl) (PVA) as a skin curative is increasing widely. However, the use of this hydrogel is challenging due to its favorable properties for microbiota growth. The association with silver nanoparticles (AgNPs) as an antimicrobial agent turns the match for PVA as a dressing, as it focuses on creating a physical barrier to avoid wound dehydration. When associated with extracellular components, such as the collagen matrix, the device obtained can create the desired biological conditions to act as a skin substitute. This study aimed to analyze the anti-microbiological activity and the in vitro and in vivo responses of a bilaminar device of PVA containing AgNPs associated with a membrane of collagen-hyaluronic acid (col-HA). Additionally, mesenchymal stem cells were cultured in the device to evaluate in vitro responses and in vivo immunomodulatory and healing behavior. The device morphology revealed a porous pattern that favored water retention and in vitro cell adhesion. Controlled wounds in the dorsal back of rat skins revealed a striking skin remodeling with new epidermis fulfilling all previously injured areas after 14 and 28 days. No infections or significant inflammations were observed, despite increased angiogenesis, and no fibrosis-markers were identified as compared to controls. Although few antibacterial activities were obtained, the addition of AgNPs prevented fungal growth. All results demonstrated that the combination of the components used here as a dermal device, chosen according to previous miscellany studies of low/mid-cost biomaterials, can promote skin protection avoiding infections and dehydration, minimize the typical wound inflammatory responses, and favor the cellular healing responses, features that give rise to further clinical trials of the device here developed.

Influence of Temperature on the Mechanical Performance of Unidirectional Carbon Fiber Reinforced Polymer Straps.

The performance of pretensioned, laminated, unidirectional (UD), carbon fiber reinforced polymer (CFRP) straps, that can potentially be used for example as bridge deck suspender cables or prestressed shear reinforcements for reinforced concrete slabs and beams, was investigated at elevated temperatures. This paper aims to elucidate the effects of elevated temperature specifically on the tensile performance of pretensioned, pin-loaded straps. Two types of tests are presented: (1) steady state thermal and (2) transient state thermal. Eight steady-state target temperatures in the range of 24 °C to 600 °C were chosen, based on results from dynamic mechanical thermal analysis (DMTA) and thermogravimetric analysis (TGA). Transient state thermal tests were performed at three sustained tensile load levels, namely 10, 15, and 20 kN, corresponding to 25%, 37%, and 50% of the ultimate tensile strength of the pin-loaded straps at ambient temperature. In general, the straps were able to retain about 50% of their ambient temperature ultimate tensile strength (UTS) at 365 °C.

Proteomic profiles and cytokeratin 13 as a potential biomarker of Ovis aries papillomavirus 3-positive and negative cutaneous squamous cell carcinomas.

Ovis aries papillomavirus 3 (OaPV3) is an epidermotropic PV reported in sheep cutaneous squamous cell carcinoma (SCC). The presence of OaPV3 DNA and its transcriptional activity in cutaneous SCC, as well as its in vitro transforming properties, suggest a viral etiology for this neoplasm. Nevertheless, the reactome associated with viral-host interaction is still unexplored. Here, we investigated and compared the proteomic profiles of OaPV3-positive SCCs, OaPV3-negative SCCs, and non-SCC samples by liquid chromatography tandem-mass spectrometry (LC-MS/MS) analysis, bioinformatics tools, and immunohistochemistry (IHC). OaPV3-positive SCCs (n = 3), OaPV3-negative SCCs (n = 3), and non-SCCs samples (n = 3) were subjected to a shotgun proteomic analysis workflow to assess protein abundance differences among the three sample classes. Proteins involved in epithelial cell differentiation, extracellular matrix organization, and apoptotic signaling showed different abundances in OaPV3-positive SCCs tissues (P ≤ 0.05) when compared to the other tissues. Cytokeratin 13 (CK 13) was among the most increased proteins in OaPV3-positive SCC and was validated by immunohistochemistry on 10 samples per class, confirming its potential as a biomarker of OaPV3 infection in SCC. Collectively, results provide a preliminary insight into the reactome associated with viral-host interaction and pave the way to the development of specific biomarkers for viral-induced sheep SCC.

Hippocampal-sparing radiotherapy and neurocognitive impairment: A systematic literature review.

INTRODUCTION: Whole-brain radiation therapy (WBRT) is an effective therapeutic modality in patients with brain metastases. However, nearly 90% of patients undergoing WBRT suffer from a neurocognitive function (NCF) impairment at diagnosis, and up to two-thirds will experience a further decline within 2-6 months after WBRT. Focal-dose reduction on bilateral hippocampus is thought to improve NCF preservation. The aim was to present a systematic review of clinical results on NCF after hippocampal-sparing (HS) WBRT. MATERIALS AND METHODS: A systematic review of published literature was performed on PubMed and the Cochrane Library. Only prospective clinical trials reporting NCF outcome in patients treated with HS-WBRT have been analyzed. RESULTS: A total of 165 patients from three studies were included. These studies are characterized by small sample size and different methods in terms of WBRT technique but with similar planning analysis and NCF assessment tests. No significant changes in NCF (i.e., verbal and nonverbal learning memory, executive functions, and psychomotor speed) between baseline and 4-month follow-up after RT and only a mean relative decline in delayed recall at 4 months (7% compared to 30% of historical control) were observed. CONCLUSIONS: Considering preliminary results on NCF preservation, further studies seem justified in patients undergoing brain irradiation for brain metastases or referred for prophylactic cranial irradiation to evaluate long-term effects on NCF and quality of life.

Intensity-Modulated Radiotherapy with Concomitant Boost After Breast Conserving Surgery: A Phase I-II Trial.

PURPOSE: A concomitant boost (CB) in patients treated with postoperative radiotherapy after conservative surgery of invasive breast cancer (BC) has been suggested for treatment time reduction and therapy intensification. The aim of this analysis was to assess long-term tolerability of a CB in patients treated with postoperative intensity Modulated Accelerated RAdiotherapy (MARA). PATIENTS AND METHODS: In this phase I-II trial, 321 patients with intermediate-high risk BC (pT1-4 with at least one of the following characteristics: pre or perimenopausal status, pN2-3, positive or close margins) were enrolled. Patients were treated with forward-planned intensity modulated radiotherapy (IMRT) and CB. A total dose of 50 Gy (2 Gy/fraction) and 60 Gy (2.4 Gy/fraction) was prescribed to the whole breast and the tumor bed, respectively. The potential impact of hypertension, diabetes, smoking habit, alcohol consumption, chemotherapy, and hormone therapy on both skin and subcutaneous late toxicity-free survival (LTFS) was evaluated. Survival curves were calculated using the Kaplan-Meier method. RESULTS: Median follow-up was 52 months (range: 3-115). Regional node irradiation, adjuvant chemotherapy and hormonal therapy were prescribed to 29.3%, 65.4% and 81.0% of patients, respectively. Five-year G2 and G3 skin LTFS were 95.6% and 100.0%, respectively. Five-year G2 and G3 subcutaneous LTFS were 80.0% and 98.6%, respectively. Only diabetes showed a significant correlation with worse G3 subcutaneous LTFS (p: 0.024). Five-year loco-regional control, metastasis-free survival, disease-free survival, and overall survival were 98.0%, 91.8%, 89.7% and 96.3%, respectively. CONCLUSION: IMRT combined with CB was associated with a low risk of > G2 late toxicities (0.0% and 1.4% for skin and subcutaneous tissue, respectively). The cumulative actuarial incidence of local recurrences was 2.0% despite the exclusion of low-risk patients. Our results suggest that CB is safe and effective in patients with intermediate-high risk BC. TRIAL REGISTRATION: ClinicalTrials.gov: NCT03471741.