A Data Augmentation Methodology to Reduce the Class Imbalance in Histopathology Images.

Deep learning techniques have recently yielded remarkable results across various fields. However, the quality of these results depends heavily on the quality and quantity of data used during the training phase. One common issue in multi-class and multi-label classification is class imbalance, where one or several classes make up a substantial portion of the total instances. This imbalance causes the neural network to prioritize features of the majority classes during training, as their detection leads to higher scores. In the context of object detection, two types of imbalance can be identified: (1) an imbalance between the space occupied by the foreground and background and (2) an imbalance in the number of instances for each class. This paper aims to address the second type of imbalance without exacerbating the first. To achieve this, we propose a modification of the copy-paste data augmentation technique, combined with weight-balancing methods in the loss function. This strategy was specifically tailored to improve the performance in datasets with a high instance density, where instance overlap could be detrimental. To validate our methodology, we applied it to a highly unbalanced dataset focused on nuclei detection. The results show that this hybrid approach improves the classification of minority classes without significantly compromising the performance of majority classes.

Rational approach to design gold nanoparticles for photothermal therapy: the effect of gold salt on physicochemical, optical and biological properties.

Among the unique characteristics associated to gold nanoparticles (AuNPs) in biomedicine, their ability to convert light energy into heat opens ventures for improved cancer therapeutic options, such as photothermal therapy (PTT). PTT relies on the local hyperthermia of tumor cells upon irradiation with light beams, and the association of AuNPs with radiation within the near infrared (NIR) range constitutes an advantageous strategy to potentially improve PTT efficacy. Herein, it was explored the effect of the gold salt on the AuNPs' physicochemical and optical properties. Mostly spherical-like negatively charged AuNPs with variable sizes and absorbance spectra were obtained. In addition, photothermal features were assessed using in vitro phantom models. The best formulation showed the ability to increase their temperature in aqueous solution up to 19 °C when irradiated with a NIR laser for 20 min. Moreover, scanning transmission electron microscopy confirmed the rearrangement of the gold atoms in a face-centered cubic structure, which further allowed to calculate the photothermal conversion efficiency upon combination of theoretical and experimental data. AuNPs also showed local retention after being locally administered in in vivo models. These last results obtained by computerized tomography allow to consider these AuNPs as promising elements for a PTT system. Moreover, AuNPs showed high potential for PTT by resulting in in vitro cancer cells' viability reductions superior to 70 % once combine with 5 min of NIR irradiation.

Preoperative Locoregional Therapy May Relate with Stemness and Distinct Transitions Between Epithelial and Mesenchymal States in Hepatocellular Carcinoma.

Background/Objectives: Locoregional therapy (LRT) might impel hepatocellular carcinoma (HCC) to exhibit different phenotypes by modulating tumoral cell adaptation. HCCs presurgically treated with LRT were studied, focusing on stemness and mesenchymal features. Methods: Clinicopathological and immunohistochemical data (Ki67, p53, EpCAM, CK19, CK7, ASMA and vimentin expression) were considered in 89 HCC nodules (30 treated with LRT; 59 non-treated), comprising 46 liver transplanted/surgically resected patients. Results: In LRT group, well and poorly differentiated tumors without fibrous encapsulation were predominant (P < 0.05) and peritumoral necroinflammation severity tended to be greater. Peritumoral Ki67 expression was higher (P < 0.05) and p53, EpCAM, CK19 and CK7 peritumoral expression was relevant after LRT, where ablated carcinomas displayed higher peritumoral CK19 expression (P < 0.05). Tumoral ASMA and vimentin expression was higher in non-LRT group (P < 0.05). In LRT group, an exclusive association between progenitor/cholangiocytic cell and mesenchymal markers expressed by tumoral cells was observed (P < 0.05): EpCAM tumoral expression associated with vimentin stromal expression; tumoral CK19 expression associated with stromal ASMA expression; tumoral CK7 expression associated with tumoral vimentin expression. Conclusion: Peritumoral cellular proliferation and expression of progenitor/cholangiocytic cell markers seem to be more frequent after LRT, with a distinctive epithelial-mesenchymal interplay and plasticity in peritumoral and tumoral compartments.

Hepatocellular carcinoma: tumor heterogeneity and recurrence after preoperative locoregional therapy.

How locoregional therapy (LRT) may change tumor compositional and functional heterogeneity, consequently contributing to treatment resistance and tumor recurrence remains largely unknown. A series of hepatocellular carcinomas (HCCs) treated with preoperative locoregional therapy (LRT) that relapsed after surgery was studied. Thirty HCCs comprising 15 patients treated with LRT prior to liver transplantation (n = 14)/surgical resection (n = 1) were studied. Five patients undergoing pre-transplant LRT, comprising 11 HCCs, presented tumor recurrence (median recurrence time = 10 months). Clinicopathological data and immunoexpression of proliferation markers (Ki67, p53), cholangiocytic/hepatic progenitor cell (HPC) markers (EpCAM)/BerEp4, CK19, CK7) and mesenchymal markers (ASMA, vimentin) were evaluated in tumoral epithelial/stromal cells and in peritumoral parenchyma. Higher grading of tumor differentiation, microvascular invasion and tumoral cell p53 expression significantly associated with recurrence (p < 0.05). Piecemeal necrosis features were more frequent in tumors that recurred (p < 0,05). Tumoral and peritumoral Ki67 expression and EpCAM, CK19 and CK7 expression in tumoral cells tended to be higher in treated tumors that recurred. Peritumoral expression of cholangiocytic/HPC markers and tumoral epithelial and stromal cellular expression of mesenchymal markers tended to be higher for tumors without recurrence. Recurrence after transplant with preoperative LRT might be associated with poor HCC differentiation, higher cellular proliferation rate, peritumoral piecemeal necrosis features and cholangiocytic/HPC phenotypes. Understanding HCC progression factors after LRT might be important for optimizing patient selection for treatment, improving surveillance after LRT and to explore synergies between LRT and systemic targeted therapies to prevent recurrence.

Personalizable AI platform for universal access to research and diagnosis in digital pathology.

BACKGROUND AND MOTIVATION: Digital pathology has been evolving over the last years, proposing significant workflow advantages that have fostered its adoption in professional environments. Patient clinical and image data are readily available in remote data banks that can be consumed efficiently over standard communication technologies. The appearance of new imaging techniques and advanced artificial intelligence algorithms has significantly reduced the burden on medical professionals by speeding up the screening process. Despite these advancements, the usage of digital pathology in professional environments has been slowed down by poor interoperability between services resulting from a lack of standard interfaces and integrative solutions. This work addresses this issue by proposing a cloud-based digital pathology platform built on standard and open interfaces. METHODS: The work proposes and describes a vendor-neutral platform that provides interfaces for managing digital slides, and medical reports, and integrating digital image analysis services compatible with existing standards. The solution integrates the open-source plugin-based Dicoogle PACS for interoperability and extensibility, which grants the proposed solution great feature customization. RESULTS: The solution was developed in collaboration with iPATH research project partners, including the validation by medical pathologists. The result is a pure Web collaborative framework that supports both research and production environments. A total of 566 digital slides from different pathologies were successfully uploaded to the platform. Using the integration interfaces, a mitosis detection algorithm was successfully installed into the platform, and it was trained with 2400 annotations collected from breast carcinoma images. CONCLUSION: Interoperability is a key factor when discussing digital pathology solutions, as it facilitates their integration into existing institutions' information systems. Moreover, it improves data sharing and integration of third-party services such as image analysis services, which have become relevant in today's digital pathology workflow. The proposed solution fully embraces the DICOM standard for digital pathology, presenting an interoperable cloud-based solution that provides great feature customization thanks to its extensible architecture.

Metabolic mitochondrial alterations prevail in the female rat heart 8 weeks after exercise cessation.

BACKGROUND: The consumption of high-caloric diets strongly contributes to the development of non-communicable diseases (NCDs), including cardiovascular disease, the leading cause of mortality worldwide. Exercise (along with diet intervention) is one of the primary non-pharmacological approaches to promote a healthier lifestyle and counteract the rampant prevalence of NCDs. The present study evaluated the effects of exercise cessation after a short period training on the cardiac metabolic and mitochondrial function of female rats. METHODS: Seven-week-old female Sprague-Dawley rats were fed a control or a high-fat, high-sugar (HFHS) diet and, after 7 weeks, the animals were kept on a sedentary lifestyle or submitted to endurance exercise for 3 weeks (6 days per week, 20-60 min/day). The cardiac samples were analysed 8 weeks after exercise cessation. RESULTS: The consumption of the HFHS diet triggered impaired glucose tolerance, whereas the HFHS diet and physical exercise resulted in different responses in plasma adiponectin and leptin levels. Cardiac mitochondrial respiration efficiency was decreased by the HFHS diet consumption, which led to reduced ATP and increased NAD(P)H mitochondrial levels, which remained prevented by exercise 8 weeks after cessation. Exercise training-induced cardiac adaptations in redox balance, namely increased relative expression of Nrf2 and downstream antioxidant enzymes persist after an eight-week exercise cessation period. CONCLUSIONS: Endurance exercise modulated cardiac redox balance and mitochondrial efficiency in female rats fed a HFHS diet. These findings suggest that exercise may elicit cardiac adaptations crucial for its role as a non-pharmacological intervention for individuals at risk of developing NCDs.

The Role of Apical Periodontitis Disease in the Development of Bisphosphonate-Related Osteonecrosis of the Jaw: An Animal Study.

BACKGROUND: Microorganisms and their by-products are responsible for establishing pulpal and periapical diseases. Healing is compromised in patients under bisphosphonate therapy, and the presence of periapical infections can potentially lead to the development of medication-related osteonecrosis of the jaw (MRONJ). This work aimed to evaluate if bisphosphonate therapy is a risk factor for MRONJ development in the presence of periapical lesions. METHODS: Two groups of 10 female Wistar rats were used. The experimental group received zoledronate (0.1 mg/kg) intraperitoneally, and the control received a saline solution, three times a week for three weeks. One week after the last injection, apical periodontitis was induced through pulpal exposure in the mandibular first molars. Twenty-one days later, the animals were intravenously injected with 99mTc-HMDP, and the radioactivity uptake by mandibular specimens was counted. In addition, sample radiographs and a histological examination were performed. RESULTS: The bone loss was higher in the control group when compared to the experimental group (p = 0.027). 99mTc-HMDP uptake in the control was reduced compared with the experimental group, although without statistical significance. CONCLUSIONS: In the presence of zoledronate therapy, apical periodontitis does not increase the risk of MRONJ development, and periapical lesions have lower bone resorption when compared to the control group.

Insights into S. aureus-Induced Bone Deformation in a Mouse Model of Chronic Osteomyelitis Using Fluorescence and Raman Imaging.

Osteomyelitis is an infection of the bone that is often difficult to treat and causes a significant healthcare burden. Staphylococcus aureus is the most common pathogen causing osteomyelitis. Osteomyelitis mouse models have been established to gain further insights into the pathogenesis and host response. Here, we use an established S. aureus hematogenous osteomyelitis mouse model to investigate morphological tissue changes and bacterial localization in chronic osteomyelitis with a focus on the pelvis. X-ray imaging was performed to follow the disease progression. Six weeks post infection, when osteomyelitis had manifested itself with a macroscopically visible bone deformation in the pelvis, we used two orthogonal methods, namely fluorescence imaging and label-free Raman spectroscopy, to characterise tissue changes on a microscopic scale and to localise bacteria in different tissue regions. Hematoxylin and eosin as well as Gram staining were performed as a reference method. We could detect all signs of a chronically florid tissue infection with osseous and soft tissue changes as well as with different inflammatory infiltrate patterns. Large lesions dominated in the investigated tissue samples. Bacteria were found to form abscesses and were distributed in high numbers in the lesion, where they could occasionally also be detected intracellularly. In addition, bacteria were found in lower numbers in surrounding muscle tissue and even in lower numbers in trabecular bone tissue. The Raman spectroscopic imaging revealed a metabolic state of the bacteria with reduced activity in agreement with small cell variants found in other studies. In conclusion, we present novel optical methods to characterise bone infections, including inflammatory host tissue reactions and bacterial adaptation.

Safety of Gold Nanoparticles: From In Vitro to In Vivo Testing Array Checklist.

In recent years, gold nanoparticles (AuNPs) have aroused the interest of many researchers due to their unique physicochemical and optical properties. AuNPs are being explored in a variety of biomedical fields, either in diagnostics or therapy, particularly for localized thermal ablation of cancer cells after light irradiation. Besides the promising therapeutic potential of AuNPs, their safety constitutes a highly important issue for any medicine or medical device. For this reason, in the present work, the production and characterization of physicochemical properties and morphology of AuNPs coated with two different materials (hyaluronic and oleic acids (HAOA) and bovine serum albumin (BSA)) were firstly performed. Based on the above importantly referred issue, the in vitro safety of developed AuNPs was evaluated in healthy keratinocytes, human melanoma, breast, pancreatic and glioblastoma cancer cells, as well as in a three-dimensional human skin model. Ex vivo and in vivo biosafety assays using, respectively, human red blood cells and Artemia salina were also carried out. HAOA-AuNPs were selected for in vivo acute toxicity and biodistribution studies in healthy Balb/c mice. Histopathological analysis showed no significant signs of toxicity for the tested formulations. Overall, several techniques were developed in order to characterize the AuNPs and evaluate their safety. All these results support their use for biomedical applications.

Antarctic toothfish Dissostichus mawsoni as a bioindicator of trace and rare earth elements in the Southern Ocean.

The Antarctic toothfish Dissostichus mawsoni is a Southern Ocean long-lived top predator which is regularly captured on an annual fishery operating in the region. By its biological and ecological characteristics, it is a potential bioindicator for the concentrations of trace and rare earth elements in the Antarctic. As these elements are mainly transferred through the diet and a deficiency or excess of these elements can lead to diverse health problems, it is important to measure their concentrations on the organisms. This study provides, for the first time, the concentration of 27 trace (major essential, minor essential and non-essential) and rare earth elements in the muscle of D. mawsoni captured in three areas of the Amundsen and Dumont D'Urville Seas (Antarctica). Major essential elements had the highest concentrations, with potassium (K) as the most concentrated, and rare earth elements the lowest. Significant differences between areas were found for most of the studied elements. No bioaccumulation nor biomagnification potential was found for the studied elements, with several elements decreasing concentrations towards larger individuals. Decreasing trends are related with the different habitats occupied by D. mawsoni through their life, suggesting that elements' concentrations in the water is determinant for the concentrations in this top predator, and/or there is a dilution effect as the fish grows. Our results also support that Se presents a detoxification potential for Hg in D. mawsoni, but only when Hg concentrations are higher to unhealthy levels. This study supports D. mawsoni as a potential bioindicator for the concentrations of the different trace and rare earth elements in the Southern Ocean, though only when comparing individuals of similar size/age, but also to evaluate annual changes on their concentrations. Furthermore, D. mawsoni can be a good source of major essential elements to humans with concentrations of major essential elements above some of other marine fish worldwide.

Validation of a methodology to quantify macro, micro, and potentially toxic elements in food matrices.

To routinely assess whether consumed food meets international guidelines, a single analytical method able to quantify minerals and potentially toxic elements (PTEs) in real food matrices is required. This work validated a simple and efficient method to quantify nine elements in different food matrices by ICP-MS. Samples from local markets (chicken, mussels, fish, rice, and seaweed) and certified reference materials were digested with HNO3 68 % and H2O2 30 %. All performance criteria (working range, linearity, LOD, LOQ, selectivity, repeatability, and trueness) met the requirements of the Portuguese Association of Accredited Laboratories. Comparison between raw and cooked food showed significant changes in most element levels, and PTEs contents complied with the maximum permissible values (EC N° 1881/2006). The minimum and maximum amounts of the foods studied, alone or in combination, raw or cooked, that can be daily consumed to meet EFSA and WHO nutritional and safety requirements were analysed.

Sexualidades amordaçadas: a atualidade dos falatórios de stella do patrocínio / Sexualidades amordazadas: la actualidad de los conversaciones de stella do patrocínio / Gagged sexualities: the topicality of stella do patrocínio's speeches

Resumo No contexto atual de retrocessos no campo da saúde mental, dos direitos sexuais e dos direitos reprodutivos, os falatórios poéticos de Stella do Patrocínio ecoam como registros de um passado manicomial que insiste em retornar. O objetivo deste artigo é problematizar os falatórios de Stella do Patrocínio, contidos no seu livro póstumo "Reino dos bichos e dos animais é o meu nome" (2001), no que se refere às sexualidades. A partir de um diálogo interseccional, tais falatórios permitiram ver uma sexualidade associada a enquadramentos normativos de gênero, raça e classe e seus efeitos nos processos de subjetivação. Além disso, foi evidente a articulação entre a lógica asilar, a biopolítica e a eugenia com os saberes e práticas das ciências psis que tendem à tutela e normalização das sexualidades, principalmente de corpos como o de Stella. Portanto, Stella nos deixa importantes contribuições e questionamentos para o campo da psicologia.

Resumen En el contexto actual de retrocesos en el campo de la salud mental, los derechos sexuales y los derechos reproductivos, las diatribas poéticas de Stella do Patrocínio resuenan como registros de un pasado asilo que insiste en volver. El objetivo de este artículo es discutir el discurso de Stella do Patrocínio, contenido en su libro póstumo "Reino dos bichos e dos animais é o meu nome" (2001), en relación a las sexualidades. A partir del diálogo interseccional, tales chismes permitieron vislumbrar una sexualidad asociada a marcos normativos de género, raza y clase y sus efectos en los procesos de subjetivación. Además, se evidenció la articulación entre la lógica del asilo, la biopolítica y la eugenesia con los saberes y prácticas de las ciencias psi que tienden a la protección y normalización de las sexualidades, principalmente de cuerpos como el de Stella. Por lo tanto, Stella nos deja importantes aportes e interrogantes para el campo de la psicología.

Abstract In the current context of setbacks in the field of mental health, sexual rights and reproductive rights, Stella do Patrocínio's poetic rants echo like records of an asylum past that insist on returning. The aims of this article is to problematize Stella do Patrocínio's gabbles, contained in her posthumous book "Reino dos bichos e dos animais é o meu nome" (2001), with regard to sexualities. Based on an intersectional dialogue, such talks allowed us to see a sexuality associated with normative frameworks of gender, race and class and their effects on the processes of subjectivation. Furthermore, the articulation between asylum logic, biopolitics and eugenics with the knowledge and practices of the psi sciences that tend to the custody and normalization of sexualities, mainly on bodies like Stella's, was evident. Therefore, Stella leaves us with important contributions and questions to the field of psychology.

Software tools and platforms in Digital Pathology: a review for clinicians and computer scientists.

At the end of the twentieth century, a new technology was developed that allowed an entire tissue section to be scanned on an objective slide. Originally called virtual microscopy, this technology is now known as Whole Slide Imaging (WSI). WSI presents new challenges for reading, visualization, storage, and analysis. For this reason, several technologies have been developed to facilitate the handling of these images. In this paper, we analyze the most widely used technologies in the field of digital pathology, ranging from specialized libraries for the reading of these images to complete platforms that allow reading, visualization, and analysis. Our aim is to provide the reader, whether a pathologist or a computational scientist, with the knowledge to choose the technologies to use for new studies, development, or research.

Thoracic Epithelioid Hemangioendothelioma: clinical demonstration and therapeutic procedures.

Epithelioid hemangioendotheliomais a low to intermediate grade malignant vascular tumors that can involve any organ. About 60-80%of patients are women, patient ages range 7 to 81 years, with a median age of 38 years. Four cases of thoracic epithelioid hemangioendotheliomas with different clinical presentation and disease progression are reported. Cases 1 and 2 are pulmonary epithelioid hemangioendotheliomas diagnosed at different advanced stages and patients died after 6 and 2,5 months of medical treatment, respectively. Case 3 corresponds to pleural epithelioid hemangioendothelioma, submit- ted to left lung decortication and pleuro-pericardial window; patient is free either from symptoms and radiographic manifestations for 10 months of follow-up. Case 4, of mediastinal epithelioid hemangioendothelioma, represented by a mass in the upper left mediastinum adherent to the aortic arch; patient underwent block excision of the mass followed by chemotherapy; subsequent recurrence 41 months later and the patient died 8 months after. The reported 4 cases reveal the heterogeneous clinical presentation of epithelioid hemangioendotheliomas with behavior in between benign and high-grade tumors, raising difficulty in either differentiating from other vascular tumors and previewing clinical outcome.

Nucleolin Overexpression Predicts Patient Prognosis While Providing a Framework for Targeted Therapeutic Intervention in Lung Cancer.

Notwithstanding the advances in the treatment of lung cancer with immune checkpoint inhibitors, the high percentage of non-responders supports the development of novel anticancer treatments. Herein, the expression of the onco-target nucleolin in patient-derived pulmonary carcinomas was characterized, along with the assessment of its potential as a therapeutic target. The clinical prognostic value of nucleolin for human pulmonary carcinomas was evaluated through data mining from the Cancer Genome Atlas project and immunohistochemical detection in human samples. Cell surface expression of nucleolin was evaluated by flow cytometry and subcellular fraction Western blotting in lung cancer cell lines. Nucleolin mRNA overexpression correlated with poor overall survival of lung adenocarcinoma cancer patients and further predicted the disease progression of both lung adenocarcinoma and squamous carcinoma. Furthermore, a third of the cases presented extra-nuclear expression, contrasting with the nucleolar pattern in non-malignant tissues. A two- to twelve-fold improvement in cytotoxicity, subsequent to internalization into the lung cancer cell lines of doxorubicin-loaded liposomes functionalized by the nucleolin-binding F3 peptide, was correlated with the nucleolin cell surface levels and the corresponding extent of cell binding. Overall, the results suggested nucleolin overexpression as a poor prognosis predictor and thus a target for therapeutic intervention in lung cancer.

Mortality due to respiratory infections: an alert study before COVID-19 pandemic.

Objective: Respiratory tract infections remain a common problem in clinical practice with high morbidity and mortality worldwide. In Portugal, pneumonia was the third leading death cause in 2018. Due to COVID-19 pandemic, there is a growing concern about the burden of respiratory diseases and preventable risk factors. The present study started before the pandemic and its aim was to determine the occurrence of pneumonia/bronchopneumonia in a postmortem series and to characterize its circumstantial context. Methods: A retrospective anatomopathological study was performed on cases with acute pneumonia/bronchopneumonia at the Medicolegal Portuguese Institute (2011-2017). Results: In an autopsy series of 737 patients, 521 were male and 675 presented comorbidities. The mean age was 63.87 ± 19.8 years. The most common acquisition site was community (65.1%), as natural death (65.5%). Concerning the manner of death, most cases (48.0%) were sudden deaths, followed by accidents (29.2%). A statistically significant association was observed between the medicolegal etiology and the place of infection acquisition, with higher prevalence of natural obitus (91.0%) in community-acquired pneumonia/bronchopneumonia versus higher prevalence of violent obitus in hospital-acquired pneumonia/bronchopneumonia (82.1%) (p < 0.001). Conclusions: Forensic anatomopathological postmortem data may contribute to better understand community and hospital pulmonary infections.

The Role of Rosmarinic Acid on the Bioproduction of Gold Nanoparticles as Part of a Photothermal Approach for Breast Cancer Treatment.

Breast cancer is a high-burden malignancy for society, whose impact boosts a continuous search for novel diagnostic and therapeutic tools. Among the recent therapeutic approaches, photothermal therapy (PTT), which causes tumor cell death by hyperthermia after being irradiated with a light source, represents a high-potential strategy. Furthermore, the effectiveness of PTT can be improved by combining near infrared (NIR) irradiation with gold nanoparticles (AuNPs) as photothermal enhancers. Herein, an alternative synthetic method using rosmarinic acid (RA) for synthesizing AuNPs is reported. The RA concentration was varied and its impact on the AuNPs physicochemical and optical features was assessed. Results showed that RA concentration plays an active role on AuNPs features, allowing the optimization of mean size and maximum absorbance peak. Moreover, the synthetic method explored here allowed us to obtain negatively charged AuNPs with sizes favoring the local particle accumulation at tumor site and maximum absorbance peaks within the NIR region. In addition, AuNPs were safe both in vitro and in vivo. In conclusion, the synthesized AuNPs present favorable properties to be applied as part of a PTT system combining AuNPs with a NIR laser for the treatment of breast cancer.

Congenital pulmonary airway malformation: A rare diagnosis in adulthood.

This paper consists of a clinical image of a complex developmental anomaly that is usually diagnosed prenatally or during childhood. Its detection in adult life is very rare, as happened in the present case.