The Nephroprotective Effect of Cornelian Cherry (Cornus mas L.) and Rowanberry (Sorbus aucuparia L.) in Gentamicin-Induced Nephrotoxicity on Wistar Rats with Emphasis on the Evaluation of Novel Renal Biomarkers and the Antioxidant Capacity in Correlation with Nitro-Oxidative Stress.

In spite of its well-known nephrotoxicity, gentamicin is nonetheless routinely used in humans and animals. However, no adjuvant treatments have been implemented to mitigate this harmful effect. Given this concern, medicinal plants represent a significant reservoir of natural antioxidants that could potentially reduce the renal oxidative stress induced by gentamicin. Therefore, the main objective of this research was to investigate the nephroprotective properties of Cornus mas and Sorbus aucuparia fruits in an experimental model of nephrotoxicity. The 3-week study was performed on male Wistar rats, which were randomly divided into six experimental groups, being subcutaneously treated with 50 mg/kg gentamicin and orally given Cornus mas and Sorbus aucuparia extracts, in doses of 40 mg/kg and 10 mg/kg, respectively. Antioxidant therapy significantly improved the nitro-oxidative stress parameters as well as the specific renal biomarkers KIM-1 and iNAG, demonstrating a considerable renal tubular protective impact. These outcomes were reinforced by biochemical and histopathological enhancements. Nevertheless, neither of the tested extracts succeeded in substantially diminishing BUN levels. Additionally, CysC did not significantly decline following extracts treatment, suggesting that the remedies did not effectively protect renal glomeruli against gentamicin stress. Future studies are required in order to determine the underlying mechanisms of these berries.

Immunomodulatory Effect of Novel Electrospun Nanofibers Loaded with Doxycycline as an Adjuvant Treatment in Periodontitis.

The immunomodulatory effect of a novel biomaterial obtained through electrospinning, based on polylactic acid (PLA) and nano-hydroxyapatite (nano-HAP), loaded with doxycycline (doxy) was evaluated in an animal model. The treatment capabilities as a local non-surgical treatment of periodontitis was investigated on the lower incisors of Wistar rats, after the induction of localized periodontitis using the ligature technique. Following the induction of the disease, the non-surgical treatment of scaling and root planing was applied, in conjunction with the application of the new material. The results of the treatment were evaluated clinically, using the tooth mobility and gingival index scores, as well as histologically. The salivary concentrations of matrix metalloproteinase 8 (MMP-8) and plasmatic concentrations of interleukin 1 (IL-1), interleukin 6 (IL-6) and tumor necrosis factor alpha (TNF-α) were also monitored. Two weeks after the ligature application, the periodontal disease was successfully induced in rats. The application of the novel biomaterial obtained through electrospinning was proven to be more effective in improving the clinical parameters, while decreasing the salivary MMP-8 and plasmatic IL-1 and TNF-α concentrations, compared to the simple scaling and root planing. Thus, the novel electrospun biomaterial could be a strong candidate as an adjuvant to the non-surgical periodontal therapy.

Inhibition of Tumor Growth and Modulation of Antioxidant Activity of Rhodoxanthin Isolated from Taxus baccata Aril against B16F10 Murine Malignant Melanoma.

Malignant melanoma is the most aggressive type of skin cancer, and due to the numerous limitations of current treatment methods, there is an urgent need to develop novel approaches for both the prevention and treatment of malignant melanoma, with research-oriented bioactive substances representing a notable first step. The current study decided to expand on previous rhodoxanthin research by investigating the possible anti-tumor effect as well as the effect on the antioxidant status in the case of murine melanoma in an experimental model. The 21-day study was carried out on female C57BL/6J mice. On the first day of the experiment, they were subcutaneously inoculated with 106 B16F10 cells and were given rhodoxanthin orally until the end of the study. Rhodoxanthin supplementation significantly reduced tumor growth (42.18%) and weight (15.74%). Furthermore, the epidermal growth factor (EGF) activity was reduced and the concentration of 8-OHdG dropped in the treated melanoma-bearing mice compared to the untreated ones, demonstrating the role of rhodoxanthin in slowing tumor growth, one of the mechanisms being the reduction of EGF level and the decrease of DNA oxidation. The administration of rhodoxanthin determined variations in antioxidant enzymes, both at the plasma level and at the tissue level.

Phytochemical Characterization of Taxus baccata L. Aril with Emphasis on Evaluation of the Antiproliferative and Pro-Apoptotic Activity of Rhodoxanthin.

Taxus baccata L., an evergreen tree, was known until recently due to its high concentration of toxic compounds. The purpose of the present study was to focus on the only non-poisonous part, the red arils, which have recently been described as an important source of various bioactive constituents. To establish total phenolic, flavonoid, and carotenoid content, antioxidant capacity, and cytotoxic properties, two types of extracts were obtained. The chemical profile of the ethanolic extract was evaluated using chromatographic (HPLC-DAD-ESI+) and spectral (UV-Vis) methods, and the antioxidant activity of the ethanolic extract was assessed using DPPH and FRAP assays, yielding moderate results. In the second type of extract (methanol: ethyl acetate: petroleum ether (1:1:1, v/v/v)) we identified three carotenoids using open column chromatography and RP-PAD-HPLC, with rhodoxanthin being the most abundant. Considering the above and mainly because of the lack of information in the literature about this pigment and its biological effects, we decided to further investigate the cytotoxic activity of rhodoxanthin, the main carotenoid presented in aril, and its protective effect against H2O2-induced oxidative stress using two cell lines: human HaCaT keratinocytes and B16F10 murine malignant melanoma. The MTT and Annexin-V Apoptosis assays showed a substantial cytotoxic potential expressed in a dose-dependent manner towards the melanoma cell line, however, no obvious cytotoxic effects on human keratinocytes were noticed.

First report of deep crypt secretory cells in ascending colon in chinchilla (Chinchilla lanigera): a histological and histochemical study / Primer reporte de células secretoras de criptas profundas en colon ascendente en chinchilla (Chinchilla lanigera): un estudio histológico e histoquímico

SUMMARY: The normal morphology of the colon differs among mammal species.The ascending colon presents several types of cells, responsible for carrying different functions for this organ. Among them, the mucus-secreting cells ensure the integrity of the mucosa, local defense, protection against different external factors, inflammatory diseases, cancer, etc. The ascending colon from 5 adult male chinchillas were processed for paraffin embedding and stained with three methods: Goldner's trichrome, PAS reaction, and Alcian blue staining procedure. The results showed that the structure of the ascending colon is similar to the one described in other species, i.e. mucosa, submucosa, muscularis externa, and serosa. Regarding the mucus-secreting cells present in the deeper part of the mucosal crypts (deep crypt secretory or DCS cells) turned out to be different not only morphologically from the surface goblet cells but also regarding the type of mucus synthesized. DCS cells have a multivacuolated, faintly stained cytoplasm with moderately PAS-positive reaction and intensely positive reaction to Alcian blue stain. The mean surface of DCS cells was 521.6 μm2 as compared to 437.9 μm2 for goblet cells (p<0.05). In conclusion, our study describes for the first time in chinchilla (Chinchilla lanigera) the presence of formerly known non-goblet or vacuolated cells, and recently entitled DCS cells in the glandular epithelium of the colon. The understanding of morphological peculiarities in chinchilla may serve as a good basis to understand the pathophysiology of various conditions that may arise.

RESUMEN: La morfología normal del colon es diferente entre las especies de mamíferos. El colon ascendente presenta varios tipos de células, encargadas de llevar varias funciones a este órgano. Entre ellos, las células secretoras aseguran la integridad de la mucosa, defensa local, protección frente a diferentes factores externos, enfermedades inflamatorias, cáncer, etc. Se procesaron para su inclusión en parafina el colon ascendente de 5 chinchillas machos adultos y se tiñeron con tres métodos: tricrómico de Goldner, reacción PAS y Azul de Alcian. Los resultados mostraron que la estructura de del colon ascendente es similar a la descrita en otras especies, es decir, mucosa, submucosa, muscular externa y serosa. Las células secretoras de la mucosa presente en la parte más profunda de las criptas mucosas (células secretoras de la cripta profunda o células DCS) resultaron ser diferentes morfológicamente de las células caliciformes superficiales, con citoplasma levemente teñido con reacción PAS positiva moderada y reacción intensamente positiva a Azul de Alcian. La superficie media de las células DCS fue de 521,6 μm2 en comparación con 437,9 μm2 de las células caliciformes (p <0,05). En conclusión, nuestro estudio describe por primera vez en chinchilla (Chinchilla lanigera) la presencia de células no caliciformes o vacuoladas anteriormente conocidas, y recientemente denominadas células DCS en el epitelio glandular del colon. La comprensión de las peculiaridades morfológicas de la chinchilla puede servir como una buena base para comprender la fisiopatología de las diversas afecciones.

Ultrasonographic Algorithm for the Assessment of Sentinel Lymph Nodes That Drain the Mammary Carcinomas in Female Dogs.

The status of sentinel lymph nodes (SLNs) is decisive in staging, prognosis, and therapeutic approach. Using an ultrasonographic examination algorithm composed of B-mode, Doppler technique, contrast-enhanced ultrasound (CEUS) and elastography, this study aimed to determine the diagnostic performance of the four techniques compared to histopathological examination. 96 SLNs belonging to 71 female dogs with mammary gland carcinomas were examined. After examinations, mastectomy and lymphadenectomy were performed. Histopathological examination confirmed the presence of metastases in 54 SLNs. The elasticity score had the highest accuracy-89.71%, identifying metastases in SLNs with 88.9.9% sensitivity (SE) and 90.5% specificity (SP), ROC analysis providing excellent results. The S/L (short axis/long axis) ratio showed 83.3% SE and 78.6% SP as a predictor of the presence of metastases in SLN having a good accuracy of 81.2%. On Doppler examination, the resistivity index(RI) showed good accuracy of 80% in characterizing lymph nodes with metastases versus unaffected ones; the same results being obtained by CEUS examination. By assigning to each ultrasonographic parameter a score (0 or 1) and summing up the scores of the four techniques, we obtained the best diagnostic performance in identifying lymph node metastases with 92.2% accuracy. In conclusion, the use of the presented algorithm provides the best identification of metastases in SLNs, helping in mammary carcinoma staging and appropriate therapeutic management.

Comparative immunohistochemical assessment of the effect of repetitive anesthesia with isoflurane and sevoflurane on rat liver / Avaliação imunohistoquímica comparativa do efeito da anestesia repetitiva com isoflurano e sevoflurano sobre o fígado de rato

Abstract Background and objectives: Inhalation anesthetics are used in human, as well as veterinary medical practice. In the present study we investigated the effect of isoflurane and sevoflurane on rat hepatocytes. Methods: A total of 40 Wistar female rats were used in this study. Animals were divided in groups of 5 rats. Groups IM, SM served as control groups. Groups I1, I2, I3 were used to study isoflurane and S1, S2, S3 for sevoflurane study. They were anesthetized 3 times, for 2 h long, at 2 days interval with a concentration of: 1.5% isoflurane (I1, I2, I3) and 2% sevoflurane (S1, S2, S3). The oxygen supply throughout the anesthesia was 1 L O2/min. Groups IM, IS, I1, S1 were sacrificed immediately after the last anesthesia. Groups I2, S2 were sacrificed 6 h after the last anesthesia, and groups I3, S3, 24 h post-anesthesia. Liver samples were harvested to highlight caspase-3 in apoptotic hepatocytes. Results: Following isoflurane administration, there were less than 1% cells in apoptosis highlighted in rat livers from groups IM, I1 and I2. At 24 h post-anesthesia (group I3), a small number of apoptotic hepatocytes was highlighted (around 3.23% cells in apoptosis), with a strictly periacinar disposition, randomly distributed in a small number of hepatic lobules. After sevoflurane administration, less than 1% apoptotic hepatocytes were identified at all control moments throughout the study. Conclusions: The results suggest that the anesthetics do not present a considerable hepatotoxicity. The comparative assessment of the two anesthetics shows that sevoflurane is superior to isoflurane.

Resumo Justificativa e objetivos: Anestésicos inalatórios são usados em humanos e também na prática médica veterinária. No presente estudo investigamos o efeito de isoflurano e sevoflurano em hepatócitos de rato. Métodos: Foram usados neste estudo 40 ratos Wistar fêmeas. Os animais foram divididos em grupos de cinco. Os grupos IM e SM serviram como controle. Os grupos I1, I2 e I3 foram usados para o estudo de isoflurano e os grupos S1, S2 e S3 para o estudo de sevoflurano. Os ratos foram anestesiados três vezes, durante duas horas em intervalos de dois dias, com uma concentração de 1,5% de isoflurano (I1, I2, I3) e 2% de sevoflurano (S1, S2, S3). O fornecimento de oxigênio durante a anestesia foi de 1 L O2/min. Os grupos IM, IS, I1 e S1 foram sacrificados imediatamente após a última anestesia. Os grupos I2 e S2 foram sacrificados seis horas após a última anestesia e os grupos I3 e S3 foram sacrificados 24 horas após a anestesia. Amostras dos fígados foram colhidas para ressaltar a caspase-3 em hepatócitos apoptóticos. Resultados: Após a administração de isoflurano, havia menos de 1% das células em apoptose em destaque nos fígados dos ratos dos grupos IM, I1 e I2. Às 24 horas após a anestesia (grupo I3), um pequeno número de hepatócitos apoptóticos foi destacado (3,23% de células em apoptose), com uma disposição estritamente periacinar, distribuídos aleatoriamente em um pequeno número de lóbulos hepáticos. Após a administração do sevoflurano, menos de 1% de hepatócitos apoptóticos foi identificado em todos os momentos de controle ao longo do estudo. Conclusões: Os resultados sugerem que os anestésicos não apresentam uma hepatotoxicidade considerável. A avaliação comparativa dos dois anestésicos mostra que sevoflurano é superior ao isoflurano.