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Background/Objectives: Chronic wounds pose a significant health concern, with their prevalence increasing due to various etiologies. The global aging population further contributes to this rise, placing a substantial burden on healthcare systems in developed countries. This work aimed to develop new therapeutic options in the form of creams and dressings based on honey, gellan gum, and hyaluronic acid for preventing and treating chronic wounds across all stages. Methods: To address this, after the formulations were developed, in vitro cytocompatibility was determined. To confirm biocompatibility, an ovine wound model was used: full-thickness excisional wounds were treated with three formulations, namely gellan gum and honey sponges (GG-HNY), gellan gum, honey and hyaluronic acid sponges (GG-HA-HNY) and a honey-based cream (cream FB002). Daily assessments, including visual evaluation and wound scoring, were conducted for 30 days. Following the study period, tissues were collected for histological analyses. Results: The macroscopic examination revealed that all therapeutic groups facilitated lesion closure. Lesion size reduction, granulation tissue disappearance, and scar tissue development were consistent across all groups, with the group receiving cream demonstrating an advanced stage of healing. All groups achieved substantial wound closure by day 30, with no significant differences. Histopathological analysis following ISO standards revealed that GG-HA-HNY had the lowest ISO score, indicating minimal reactivity and inflammation, which corroborated the cytocompatibility. Conclusions: Combining these insights with previous findings enhances our understanding of wound regeneration dynamics and contributes to refining therapeutic strategies for chronic wounds. The formulations were designed to balance therapeutic efficacy with cost-effectiveness, leveraging low-cost raw materials and straightforward production methods. [ABSTRACT FROM AUTHOR] more...

Background: Stem cells are capable of self-renewal and differentiation into various specialized cells, making them a potential therapeutic option in regenerative medicine. This study establishes a comprehensive methodology for isolating, culturing, and characterizing rat hair follicle stem cells. Methods and Results: Hair follicles were harvested from Sprague–Dawley rats and subjected to two different isolation techniques. Immunohistochemical analysis and real-time PCR confirm the expression of specific surface markers and genes, validating the cells' identity. Growth kinetics, colony formation units (CFU), and tri-differentiation capacity were also assessed. Additionally, the cells' secretome was analyzed, regarding its content in biofactors with wound healing properties. Conclusions: These findings highlight the potential of these cells as a valuable cell source for skin regeneration applications. They contribute to advancing our understanding of stem cell applications in regenerative medicine and hold promise for therapeutic interventions in various clinical contexts, aligning with broader research on the diverse capabilities of hair follicle stem cells. [ABSTRACT FROM AUTHOR] more...

Breast cancer epithelial cells with the CD44+/CD24-/low phenotype possess tumor-initiating cells and epithelial-mesenchymal transition (EMT) capacity. Massive parallel sequencing can be an interesting approach to deepen the molecular characterization of these cells. We characterized CD44+/CD24-/cytokeratin(Ck)+/CD45- cells isolated through flow cytometry from 43 biopsy and 6 mastectomy samples harboring different benign and malignant breast lesions. The Ion Torrent Ampliseq Cancer Hotspot panel v2 (CHPv2) was used for the identification of somatic mutations in the DNA extracted from isolated CD44+/CD24-/Ck+/CD45- cells. E-Cadherin and vimentin immunohistochemistry was performed on sections from the corresponding formalin-fixed, paraffin-embedded (FFPE) blocks. The percentage of CD44+/CD24-/Ck+/CD45- cells increased significantly from non-malignant to malignant lesions and in association with a significant increase in the expression of vimentin. Non-malignant lesions harbored only a single-nucleotide polymorphism (SNP). Mutations in the tumor suppressor p53 (TP53), NOTCH homolog 1 (NOTCH1), phosphatase and tensin homolog (PTEN), and v-akt murine thymoma viral oncogene homolog 1 (AKT1) genes were found in isolated CD44+/CD24-/Ck+/CD45- cells from ductal carcinomas in situ (DCIS). Additional mutations in the colony-stimulating factor 1 receptor (CSF1R), ret proto-oncogene (RET), and TP53 genes were also identified in invasive ductal carcinomas (IDCs). The use of massive parallel sequencing technology for this type of application revealed to be extremely effective even when using small amounts of DNA extracted from a low number of cells. Additional studies are now required using larger cohorts to design an appropriate mutational profile for this phenotype. [ABSTRACT FROM AUTHOR] more...

Introduction: Degenerative myelopathy (DM) is a neurodegenerative spinal cord disease with upper motor neurons, with progressive and chronic clinical signs, similar to amyotrophic lateral sclerosis (ALS). DM has a complex etiology mainly associated with SOD1 gene mutation and its toxic role, with no specific treatment. Daily intensive rehabilitation showed survival time near 8 months but most animals are euthanized 6-12 months after clinical signs onset., Methods: This prospective controlled blinded cohort clinical study aims to evaluate the neural regeneration response ability of DM dogs subjected to an intensive neurorehabilitation protocol with mesenchymal stem cells (MSCs) transplantation. In total, 13 non-ambulatory (OFS 6 or 8) dogs with homozygous genotype DM/DM and diagnosed by exclusion were included. All were allocated to the intensive neurorehabilitation with MSCs protocol (INSCP) group ( n = 8) or to the ambulatory rehabilitation protocol (ARP) group ( n = 5), which differ in regard to training intensity, modalities frequency, and MSCs transplantation. The INSCP group was hospitalized for 1 month (T0 to T1), followed by MSCs transplantation (T1) and a second month (T2), whereas the ARP group was under ambulatory treatment for the same 2 months., Results: Survival mean time of total population was 375 days, with 438 days for the INSCP group and 274 for the ARP group, with a marked difference on the Kaplan-Meier survival analysis. When comparing the literature's results, there was also a clear difference in the one-sample t -test ( p = 0.013) with an increase in time of approximately 70%. OFS classifications between groups at each time point were significantly different ( p = 0.008) by the one-way ANOVA and the independent sample t -test., Discussion: This INSCP showed to be safe, feasible, and a possibility for a long progression of DM dogs with quality of life and functional improvement. This study should be continued., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2023 Gouveia, Correia, Cardoso, Carvalho, Oliveira, Almeida, Gamboa, Ribeiro, Branquinho, Sousa, Lopes, Sousa, Moreira, Coelho, Rêma, Alvites, Ferreira, Maurício and Martins.) more...

The persistent organic pollutant p,p'-DDE, the major metabolite of the insecticide DDT, has displayed evidence of endocrine disruption through the inhibition of androgen binding to androgen receptors in different species. Although p,p'-DDE was continuously detected in wild fish with abnormal gonad development such as intersex, little is known about its mode of action during gonad development in fish. To elucidate the potential endocrine effects of this pollutant in zebrafish (Danio rerio), juveniles (30 days post hatch) were exposed to p,p'-DDE during the critical window of sexual differentiation. Fish were exposed to sublethal concentrations ranging from 0.01 to 20 µg l(-1) over 14 days and were maintained in control water for an additional 4 months. As core endpoints, the vitellogenin (vtg) concentration was measured at the end of exposure, and sex ratio and the gonadosomatic index were assessed 4 months after the end of exposure. An increase in vtg production in whole body homogenate was observed in fish exposed to 0.2 and 2.0 µg l(-1) p,p'-DDE. No significant differences were displayed in morphological parameters such as the gonadosomatic index of males and females or sex ratio. However, exposed females presented histopathological changes that include the reduction of the number of mature oocytes, which might impair their successful reproduction. These results demonstrate the ability of p,p'-DDE to cause endocrine disruption in zebrafish exposed during gonad differentiation of juvenile specimens. Furthermore, vtg induction by p,p'-DDE in juvenile zebrafish arises as a predictive marker for adverse effects of this DDT metabolite on the ovarian function of female zebrafish., (Copyright © 2014 John Wiley & Sons, Ltd.) more...

Background/aim: Claudins (CLDNs) are crucial structural and functional components of tight junctions playing an important role in maintaining cell polarity, controlling paracellular diffusion and regulating cell growth and differentiation of epithelial cells. CLDNs are differentially expressed in neoplastic lesions compared to the corresponding healthy tissues; therefore, they are thought to play a role in tumorigenesis and tumor progression. Reduced expression of CLDN-1 has been observed in several types of human cancers, including thyroid tumors. There are no reports of CLDN-1 immunoexpression in normal and neoplastic canine thyroid tissues., Material and Methods: CLDN-1 immunoexpression was investigated in normal canine thyroid gland (n=2), benign (n=1) and malignant (n=10) tumors, as well as neoplastic emboli (n=6)., Results: CLDN-1 was constitutively expressed in normal canine follicular epithelium. Ninety percent of the malignant canine thyroid lesions showed absence or reduced CLDN-1 expression compared to that of normal thyroid gland. Additionally, a cytoplasmic subcellular location of CLDN-1 was recorded in the malignant epithelial cells and neoplastic emboli., Conclusion: These findings link altered expression of CLDN-1 to neoplastic transformation and suggest that CLDN-1 expression is associated with malignant canine thyroid tumors and their vascular invasion., (Copyright © 2015 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved.) more...

Background/aim: The Cancer Stem Cell (CSC) model proposes that cancer is driven by a cellular component which possesses stem cell (SC) properties, cancer stem cells (CSCs), a distinct cell-type which is tumorigenic and capable of invasion and metastasis. Enzymatic activity of aldehyde dehydrogenase-1 (ALDH1), a de-toxifying enzyme that oxidizes intracellular aldehydes, has been used as a marker of normal and malignant breast stem cells (BSCs). CD44-transmembrane protein has already been shown to possess the ability to identify breast epithelial cells with stem properties., Materials and Methods: In order to compare two of the currently most reliable BSCs markers, ALDH1 and CD44, and to correlate their expression within different breast lesions, 190 samples from breast cancer specimens were analyzed by tissue microarrays., Results: ALDH1 expression was observed in 85.43% and CD44 in 90.3% of all samples. No overexpression was observed for ALDH1 between invasive tumors, ductal carcinomas in situ and non-malignant lesions of breast, although ALDH1 had a significant negative correlation with estrogen-receptor (ER) and progesterone-receptor (PR) status (p=0.002 and p=0.001, respectively) and a positive correlation with CD44 (p<0.001). Moreover, combined overexpression of ALDH1 and CD44 was observed in ductal in situ tumors (p<0.001)., Conclusion: The combined overexpression of these markers in ductal carcinomas in situ is in agreement with the CSC model in breast cancer. more...