Histological changes in the vulva and vagina from ovariectomised rats undergoing oestrogen treatment

Background: The purpose of this study was to assess the histological changes occurring in the vagina and vulva in ovariectomised female rats, as well as the response to the administration of injectable oestrogens. Material and methods: We used 30 female Wistar white rats, distributed as follows: group 1 — the control group, group 2 — the operated but untreated rats, and groups 3, 4 and 5 — operated rats, to which oestrogenic treatment was administered (Estradiol, Estradurin, Sintofolin) at a dosage of 0.2 mg/rat/day. After 14 days of treatment, all animals were sacrificed and vaginal and vulvar biopsies were taken from all groups. Results: In group 2, we encountered structural changes of the vaginal mucosa, with severe atrophy and alterations in the thickness of the vagina and vulva. In groups 3, 4 and 5 we found marked hyperplasia of the vaginal and vulvar epithelium, eosinophilic and mast cell infiltration in the chorion. Conclusions: Our study proves that the histopathological changes during anoestrus after administration of oestrogens are cell hyperplasia, thickening of the superficial mucosal layer, eosinophilic and mast cells infiltrations, and chorionic congestion. Furthermore, we demonstrated that Estradiol therapy induces the most evident histological changes when compared to synthetic oestrogens such as Estradurin or Sintofolin.

The stem cells in chronic experimental liver diseases

Introduction: Chronic liver diseases (CLD) are increasing worldwide affecting almost 17% of general population."In the Republic of Moldova during the last ten years the incidence and prevalence of CLD had increased continuously. The liver cirrhosis in our country is the third cause of death after cardiovascular diseases and cancer. Ihis situation is the result of a high prevalence of infectious viral hepatitis, alcohol and hepatotoxic drug abuse, and the unavailable so far of the orthotopic liver transplantation (OLT), the only curative treatment for rhe end stage liver diseases. The number of patients waiting for an OLT has increased during the last years, meanwhile the organ donation has not kept up with demand. Consequently the organ shortage is increasing the morbidity and mortality of patients on waiting list. This clearly implies the need for finding alternative solutions for the patients with end stage liver disease, and stem ceil therapy is the one that gives the most hope so far. The aim ot this study was to induce chronic experimental liver disease in rats, then transplant stem cells and further evaluate the effect on liver function. Material and methods: Chronic liver lesions were induced on white female rats of 6-8 months age, weighting 210-250 mg, by injecting CCL4 subcutaneously, dissolved in olive oil, twice a week, for 8 weeks. At the end of 6 weeks the rats were divided into 5 groups with further intrasplenic transplantation of 6xl03 allogenic stem cells (SC) performed. The animals in group 1 received blood marrow SC, the second group received umbilical cord SC, the third group received hepatic fetal SC. The fourth and fifth groups received intrasplenic saline solution only. Meanwhile we continue to inject CCL4 subcutaneously twice a week for the groups 1,2,3 to prevent endogenous liver regeneration and allow the stem cells to act. For the fourth group we continue with CCL4 and for the 5 without CCL4 to allow endogenous regeneration for another 6 weeks. The animals were sacrificed at 10, 20 and 40 days after transplantation, and there were collected 5 ml of blood and the liver specimens. Preventive results: After 6 weeks of CCL4 administration 90% of rats presented weight loss ranging between 5 to 20%, and signs of coagulopathy like periocular bleeding. The 6 rats sacrificed just before the SC transplantation proved the presence of ascites and yellow, nodular liver changes. Histological examination showed the presence of infiltration of the liver with neutrophils, regenerating nodules of hepatocytes and the deposition of connective tissue between these nodules. Conclusions: Further biochemical, histological and immunohystochemical analyses have to be done on liver specimen and collected blood to evaluate the effects of SC therapy on the end stage of the liver disease

Selective ex-vivo photothermal ablation of human pancreatic cancer with albumin functionalized multiwalled carbon nanotubes

The process of laser-mediated ablation of cancer cells marked with biofunctionalized carbon nanotubes is frequently called “nanophotothermolysis”. We herein present a method of selective nanophotothermolisys of pancreatic cancer (PC) using multiwalled carbon nanotubes (MWCNTs) functionalized with human serum albumin (HSA). With the purpose of testing the therapeutic value of these nanobioconjugates, we have developed an ex-vivo experimental platform. Surgically resected specimens from patients with PC were preserved in a cold medium and kept alive via intra-arterial perfusion. Additionally, the HSA-MWCNTs have been intra-arterially administered in the greater pancreatic artery under ultrasound guidance. Confocal and transmission electron microscopy combined with immunohistochemical staining have confirmed the selective accumulation of HSA-MWCNTs inside the human PC tissue. The external laser irradiation of the specimen has significantly produced extensive necrosis of the malign tissue after the intra-arterial administration of HSA-MWCNTs, without any harmful effects on the surrounding healthy parenchyma. We have obtained a selective photothermal ablation of the malign tissue based on the selective internalization of MWCNTs with HSA cargo inside the pancreatic adenocarcinoma after the ex-vivo intra-arterial perfusion

Enhanced laser thermal ablation for the in vitro treatment of liver cancer by specific delivery of multiwalled carbon nanotubes functionalized with human serum albumin

The main goal of this investigation was to develop and test a new method of treatment for human hepatocellular carcinoma (HCC). We present a method of carbon nanotube-enhanced laser thermal ablation of HepG2 cells (human hepatocellular liver carcinoma cell line) based on a simple multiwalled carbon nanotube (MWCNT) carrier system, such as human serum albumin (HSA), and demonstrate its selective therapeutic efficacy compared with normal hepatocyte cells. Both HepG2 cells and hepatocytes were treated with HSA–MWCNTs at various concentrations and at various incubation times and further irradiated using a 2 W, 808 nm laser beam. Transmission electron, phase contrast, and confocal microscopy combined with immunochemical staining were used to demonstrate the selective internalization of HSA–MWCNTs via Gp60 receptors and the caveolin-mediated endocytosis inside HepG2 cells. The postirradiation apoptotic rate of HepG2 cells treated with HSA–MWCNTs ranged from 88.24% (for 50 mg/L) at 60 sec to 92.34% (for 50 mg/L) at 30 min. Significantly lower necrotic rates were obtained when human hepatocytes were treated with HSA–MWCNTs in a similar manner. Our results clearly show that HSA–MWCNTs selectively attach on the albondin (aka Gp60) receptor located on the HepG2 membrane, followed by an uptake through a caveolin-dependent endocytosis process. These unique results may represent a major step in liver cancer treatment using nanolocalized thermal ablation by laser heating

Effects of processing on polyphenolic and volatile composition and fruit quality of clery strawberries

Strawberries belonging to cultivar Clery (Fragaria x ananassa (Duchesne ex Weston)), cultivated in central Italy were subjected to a multi‐methodological experimental study. Fresh and defrosted strawberries were exposed to different processing methods, such as homogenization, thermal and microwave treatments. The homogenate samples were submitted to CIEL*a*b* color analysis and Head‐Space GC/MS analysis to determine the impact of these procedures on phytochemical composition. Furthermore, the corresponding strawberry hydroalcoholic extracts were further analyzed by HPLC‐DAD for secondary metabolites quantification and by means of spectrophotometric in vitro assays to evaluate their total phenolic and total flavonoid contents and antioxidant activity. These chemical investigations confirmed the richness in bioactive metabolites supporting the extraordinary healthy potential of this fruit as a food ingredient, as well as functional food, highlighting the strong influence of the processing steps which could negatively impact on the polyphenol composition. Despite a more brilliant red color and aroma preservation, nonpasteurized samples were characterized by a lower content of polyphenols and antioxidant activity with respect to pasteurized samples, as also suggested by the PCA analysis of the collected data

Helicobacter pullorum cytolethal distending toxin targets vinculin and cortactin and triggers formation of lamellipodia in intestinal epithelial cells

Helicobacter pullorum, a bacterium initially isolated from poultry, has been associated with human digestive disorders. However, the factor responsible for its cytopathogenic effects on epithelial cells has not been formally identified. The cytopathogenic alterations induced by several human and avian H. pullorum strains were investigated on human intestinal epithelial cell lines. Moreover, the effects of the cytolethal distending toxin (CDT) were evaluated first by using a wild-type strain and its corresponding cdtB isogenic mutant and second by delivering the active CdtB subunit of the CDT directly into the cells. All of the H. pullorum strains induced cellular distending phenotype, actin cytoskeleton remodeling, and G2/M cell cycle arrest. These effects were dependent on the CDT, as they were (1) not observed in response to a cdtB isogenic mutant strain and (2) present in cells expressing CdtB. CdtB also induced an atypical delocalization of vinculin from focal adhesions to the perinuclear region, formation of cortical actin-rich large lamellipodia with an upregulation of cortactin, and decreased cellular adherence. In conclusion, the CDT of H. pullorum is responsible for major cytopathogenic effects in vitro, confirming its role as a main virulence factor of this emerging human pathogen.This work was supported by the Institut national de la santé et de la recherche médicale, the University Bordeaux Segalen, the Conseil Régional d’Aquitaine (grants 20030304002FA and 20040305003 FA), the Société Nationale Française de Gastroentérologie, the European Union (FEDER no. 2003227

Exploring the phytochemical profile of Cytinus hypocistis (L.) L. as a source of health-promoting biomolecules behind its in vitro bioactive and enzyme inhibitory properties

Cytinus hypocistis whole plant and its three different parts (petals, stalks, and nectar) were chemically characterised and their biological properties evaluated. A total of 17 phenolic compounds were identified, being galloyl-bis-HHDP-glucose the most abundant. All the tested extracts showed high antioxidant capacity, with the petals exhibiting the most promising results both in the OxHLIA (IC50 = 0.279 ng/mL) and TBARS (IC50=0.342 ng/mL) assays. For the antidiabetic and anti-tyrosinase enzyme inhibitory assays, the stalk extract presented the lowest IC50 values, 0.039 mg/mL and 0.09 mg/mL, respectively. Regarding antibacterial activity, all tested extracts displayed broad-spectrum microbial inhibition against both Gram-positive and Gram-negative bacteria. Similarly, all extracts displayed effective anti-proliferation activity against four tested tumour cell lines (NCI–H460, HeLa, HepG2, and MCF-7), with no toxicity observed for a non-tumour cell line. Considering the anti-inflammatory activity, the petals showed the highest nitric oxide inhibition (IC50 = 127 μg/mL). These results point C. hypocistis as a promising source of health-promoting biomolecules.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and European Regional Development Fund (FEDER) under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019). L. Barros, J. Pinela, M.I. Dias and R.C. Calhelha thank the national funding by FCT, P.I., through the institutional scientific employment program-contract for their contracts. The authors are also grateful to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E and TRANSCoLAB 0612_TRANS_CO_LAB_2_P.info:eu-repo/semantics/publishedVersio

Compositional features and bioactive properties of aloe vera leaf (Fillet, mucilage, and rind) and flower

This work aimed to characterize compositional and bioactive features of Aloe vera leaf (fillet, mucilage, and rind) and flower. The edible fillet was analysed for its nutritional value, and all samples were studied for phenolic composition and antioxidant, anti-inflammatory, antimicrobial, tyrosinase inhibition, and cytotoxic activities. Dietary fibre (mainly mannan) and available carbohydrates (mainly free glucose and fructose) were abundant macronutrients in fillet, which also contained high amounts of malic acid (5.75 g/100 g dw) and -tocopherol (4.8 mg/100 g dw). The leaf samples presented similar phenolic profiles, with predominance of chromones and anthrones, and the highest contents were found in mucilage (131 mg/g) and rind (105 mg/g) extracts, which also revealed interesting antioxidant properties. On the other hand, the flower extract was rich in apigenin glycoside derivatives (4.48 mg/g), effective against Pseudomonas aeruginosa (MIC = 0.025 mg/mL and MBC = 0.05 mg/mL) and capable of inhibiting the tyrosinase activity (IC50 = 4.85 mg/mL). The fillet, rind, and flower extracts also showed a powerful antifungal activity against Aspergillus flavus, A. niger, Penicillium funiculosum, and Candida albicans, higher than that of ketoconazole. Thus, the studied Aloe vera samples displayed high potential to be exploited by the food or cosmetic industries, among others.The authors are grateful to the Foundation for Science and Technology (FCT, Portugal) and FEDER(European Regional Development Fund) under Programme PT2020 for financial support to CIMO (UID/AGR/00690/2019) and the research contracts of J. Pinela, R.C. Calhelha, and L. Barros (national funding by FCT, through the institutional scientific employment program-contract); to the Project AllNat—POCI-01-0145-FEDER-030463 (PTDC/EQU-EPQ/30463/2017), funded by FEDER funds through COMPETE2020—Programa Operacional Competitividade e Internacionalização (POCI)—and by national funds through FCT/MCTES; and to FEDER-Interreg España-Portugal programme for financial support through the project 0377_Iberphenol_6_E.info:eu-repo/semantics/publishedVersio