This study unravels a formulation made of food‐based microparticles (MPs) able to control the release of quercetin, a natural anticancer compound, which activity is only limited by its poor aqueous ...solubility and consequent low bioavailability. To solve this issue, a spray‐dried micro delivery system was developed using a bench mini spray dryer B290 Buchi. The resulting MPs were only manufactured with food‐derived ingredients such as whey proteins and milk, avoiding the use of any other synthetic material. These microparticles were characterised with a testing campaign encompassing either the physical–chemical characterisation with SEM, DSC and DLS, or the technological and biological features with in vitro, ex vivo and in vivo studies, the latter being characterised by a human colon cancer xenograft model. These studies showed as the quercetin solubility and release rate improved when tested in condition mimicking oral administration, resulting in a general improvement of its bioavailability and the consequent anticancer activity. This study shows as the whey proteins may serves as natural adjuvant able to provide valuable technological features when used to manufactures micro carriers by spray drying.
Multiple oral administration of quercetin embodied in the microparticles prevents the tumour mass from increasing in volume compared to pure quercetin.
Betamethasone is a glucocorticoid authorised in cattle for the treatment of metabolic and inflammatory diseases, but, in Europe, it is illegally employed to improve productive performances. LC-MS/MS ...is the official control method of veterinary drugs residues in food of animal origin. An experimental study was developed to evaluate the feasibility of proton magnetic resonance spectroscopy (1H-MRS) as a potential alternative approach to detect the presence of betamethasone residues. Eight rat liver samples were collected 24 h post-betamethasone-treatment from experimental and control animals and were analysed by 1H-MRS using a 7-Tesla MRI scanner. 1H-MR reference spectra both of the Bentelan formulation used for treatment, and of three solutions of betamethasone in dimethyl sulphoxide (DMSO) at 5, 10 and 100 mM, respectively, were acquired to fit analyte-peaks in the liver samples spectra. Betamethasone-peaks were found only in the 100 mM betamethasone in DMSO solution spectrum. Betamethasone residues were not detected in any of the tissue samples analysed, probably related to the low concentration of injected drug. These findings allow us to establish, for the first time in the literature, the detection limit (in the range 10-100 mM) of betamethasone for the 7-Tesla MRI scanner used here. Given this very-low sensitivity, we conclude that the evaluated 1H-MR spectroscopy approach is not suitable for the detection of betamethasone residues in edible tissues, since the maximum residue limit imposed by Commission Regulation (EC) 37/2010 for betamethasone in the liver, and metabolic concentrations required to be detected in animal samples from livestock, are far below the detection limit we found.
Regeneration and recovery of nerve tissues are a great challenge for medicine, and positively affect the quality of life of patients. The development of tissue engineering offers a new approach to ...the problem with the creation of multifunctional artificial scaffolds that act on various levels in the damaged tissue, providing physical and biochemical support for the growth of nerve cells. In this study, the effects of the use of a tubular scaffold made of polybutylene succinate (PBS), surgically positioned at the level of a sciatic nerve injured in rat, between the proximal stump and the distal one, was investigated. Scaffolds characterization was carried out by scanning electron microscopy and X‐ray microcomputed tomography and magnetic resonance imaging, in vivo. The demonstration of the nerve regeneration was based on the evaluation of electroneurography, measuring the weight of gastrocnemius and tibialis anterior muscles, histological examination of regenerated nerves and observing the recovery of the locomotor activity of animals. The PBS tubular scaffold minimized iatrogenic trauma on the nerve, acting as a directional guide for the regenerating fibers by conveying them toward the distal stump. In this context, neurotrophic and neurotropic factors may accumulate and perform their functions, while invasion by macrophages and scar tissue is hampered.
Hepatocellular carcinoma (HCC) is a highly malignant tumor that responds very poorly to existing therapies, most probably due to its extraordinary inter‐ and intra‐tumor molecular heterogeneity. The ...modest therapeutic response to molecular targeted agents underlines the need for new therapeutic approaches for HCC. In our study, we took advantage of well‐characterized human HCC cell lines, differing in transcriptomic subtypes, DNA mutation and amplification alterations, reflecting the heterogeneity of primary HCCs, to provide a preclinical evaluation of the specific heat shock protein 90 (HSP90) inhibitor AUY922 (luminespib). Indeed, HSP90 is highly expressed in different tumor types, but its role in hepatocarcinogenesis remains unclear. Here, we analyzed HSP90 expression in primary human HCC tissues and evaluated the antitumor effects of AUY922 in vitro as well as in vivo. HSP90 expression was significantly higher in HCC tissues than in cirrhotic peritumoral liver tissues. AUY922 treatment reduced the cell proliferation and viability of HCC cells in a dose‐dependent manner, but did not do so for normal human primary hepatocytes. AUY922 treatment led to the upregulation of HSP70 and the simultaneous depletion of HSP90 client proteins. In addition, in a cell type‐dependent manner, treatment induced either both caspase‐dependent β‐catenin cleavage and the upregulation of p53, or Mcl‐1 expression, or NUPR1 expression, which contributed to the increased efficacy of, or resistance to, treatment. Finally, in vivo AUY922 inhibited tumor growth in a xenograft model. In conclusion, HSP90 is a promising therapeutic target in HCC, and AUY922 could be a drug candidate for its treatment.
What's new?
Hepatocellular carcinoma (HCC) exhibits vast molecular heterogeneity, suggesting that therapies aimed against targets that interact with multiple signaling pathways could be key to improving HCC outcome. Here, heat shock protein 90 (HSP90), a modulator of numerous signaling components, was found to be highly expressed in HCC. AUY922, an HSP90 inhibitor, blocked HCC cell growth in vitro and, in HepG2 liver carcinoma cells, induced apoptosis via caspase activation and β‐catenin fragmentation. These effects were not observed in Huh7 or SNU475 HCC cell lines. AUY922 also reduced tumor growth in vivo, marking HSP90 as a promising therapeutic target in HCC.
Hepatocellular carcinoma (HCC) is one of the common malignancies and is an increasingly important cause of cancer death worldwide. Surgery, chemotherapy, and radiation therapy extend the 5-year ...survival limit in HCC patients by only 6%. Therefore, there is a need to develop new therapeutic approaches for the treatment of this disease. The orally bioavailable proteasome inhibitor MLN2238 (ixazomib) has been demonstrated to have anticancer activity. In the present study, we investigated the preclinical therapeutic efficacy of MLN2238 in HCC cells through in vitro and in vivo models, and examined its molecular mechanisms of action. MLN2238 inhibited cell viability in human HCC cells HepG2, Hep3B, and SNU475 in a time- and dose-dependent manner. Flow cytometry analysis demonstrated that MLN2238 induced G2/M cell cycle arrest and cellular apoptosis in HCC cells. Cell cycle arrest was associated with increased expression levels of p21 and p27. MLN2238-induced apoptosis was confirmed by caspase-3/7 activation, PARP cleavage and caspase-dependent β-catenin degradation. In addition, MLN2238 activated ER stress genes in HCC cells and increased the expression of the stress-inducible gene nuclear protein-1. Furthermore, MLN2238 treatment induced upregulation of myeloid cell leukemia-1 (Mcl-1) protein, and Mcl-1 knockdown sensitized HCC cells to MLN2238 treatment, suggesting the contribution of Mcl-1 expression to MLN2238 resistance. This result was also confirmed using the novel Mcl-1 small molecule inhibitor A1210477. Association of A1210477 and MLN2238 determined synergistic antitumor effects in HCC cells. Finally, in vivo orally administered MLN2238 suppressed tumor growth of Hep3B cells in xenograft models in nude mice. In conclusion, our results offer hope for a new therapeutic opportunity in the treatment of HCC patients.
Background
Preclinical training in microsurgery usually proceeds through steps of increasing difficulty. Traditionally, advanced training is carried out on living animal models which best reproduce ...the clinical scenario, but recently, the increasing interest in animal rights has led to a greater development and spread of different non-living models for all steps of training.
Methods
The aim of this study was to identify, through a review of the literature, the inanimate models suitable for basic and intermediate/advanced training courses and to evaluate their pros and cons. The search was carried out exclusively through the PubMed database, with "microsurgery" or “supermicrosurgery” and ("training" or "non-living model") as keywords in the "title and/or abstract" fields. The filters used were: publication date (2010–2022) and species (other animals). The study was done following the PRISMA 2020 checklist criteria.
Results
A total number of 398 articles were initially screened. Following abstract review, 75 articles were selected, and 51 articles were chosen following full text review. Several non-living models are available for training on fine dissection and microsurgical technique. Among the non-animal models, food and synthetic materials (silicone tubes and latex gloves) were predominantly used. Among the non-living animals, the chicken was the most frequently used animal followed by the pig and the rat. Non-living animal mainly focus on vascular sutures on vessels of different vessels, including very small vessels for supermicrosurgery.
Conclusions
The results of this study have shown that many different non-living models are available not only for basic microsurgical training, but also for intermediate training. These models allow to improve microsurgical and supermicrosurgical skills, simultaneously reducing the use of living animals, according to the “3 R” principle. Their main limitation is that due to their characteristics, as tissue consistency and the absence of a pumping flow, they do not provide a realistic experience as that on living animals, which are still the reference for the final phases of microsurgical training.
Level of evidence:Not ratable.
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A hyaluronic acid (HA) derivative bearing octadecylamine and acylhydrazine functionalities has been here employed for the production of a paclitaxel delivering matrix for locoregional ...chemotherapy. Through a strategy consisting in a powder compression and a plasticization with a mixture water/ethanol, a physically assembled biomaterial, stable in solutions with physiologic ionic strengths, has been produced. Two different drug loading strategies have been adopted, by using paclitaxel as chemotherapic agent, and obtained samples have been assayed in terms of release in enhanced solubility conditions and in vitro and in vivo tumoural cytotoxicity. In particular sample with the best releasing characteristics was chosen for an in vivo evaluation against a HCT-116 xenograft on mice. Local tumour establishment and metastatic diffusion was assayed locally at the site of xenograft implantation and at the tributary lymph nodes. Obtained results demonstrated how loading procedure influenced paclitaxel crystallinity into the matrix and consequently drug diffusion and its cytoreductive potential. Chosen paclitaxel loaded matrix was able to drastically inhibit HCT-116 establishment and metastatic diffusion.
The present study hypothesizes that raw materials used in bread making can transfer antibiotic resistance genes (ARGs) to processed breads. Four types of flour and four types of semolina were ...purchased from supermarkets and inoculated with a commercial dried sourdough starter to make breads. The microbiological characteristics of all raw materials and fermented doughs were investigated. The levels of yeasts and lactic acid bacteria (LAB) increased up to 107 CFU/g. The values of pH decreased to 4.54–4.86 while total titratable acidity increased inversely. All unprocessed and processed samples, including breads, were analyzed by a molecular approach to detect bacterial and fungal DNAs and 17 antibiotic resistance genes for penicillins, macrolides, tetracyclines, and chloramphenicol. Illumina technology showed that the operational taxonomy units (OTUs) identified from unprocessed wheat milling products, fermented doughs, and baked products mainly belonged to Acetobacteraceae. Enterococci were present in all doughs. After baking, the relative abundance (RA)% of Enterococcus and Acetobacteraceae decreased. The DNA analyzed for fungal composition showed that Kazachstania humilis dominated dried sourdough starter and doughs, and its OTUs were also detected at high RA% in baked products. The search for ARGs revealed that all samples analyzed did not show resistance to penicillins, chloramphenicol, and macrolides. However, three of the semolinas included in this study (S1, S3 and S4) and the corresponding doughs (SD1, SD3 and SD4) were positive for tet(A) and tet(B) resistance genes. This work indicated that breads have a limited role in the dissemination of ARGs.
Inguinal hernia is a degenerative disease occurring in a high motile surround. Stopping degeneration and promoting tissue regeneration should be the treatment goal. Groin hernias are conventionally ...managed with static flat meshes, mostly fixated to the delicate inguinal environment. Far from a regenerative effect, the biologic response of conventional hernia meshes is characterized by a foreign body reaction leading to a stiff/shrunken scar plate, which is often the source of unpleasant complications. Recently, a newly engineered 3D device for inguinal hernia repair – ProFlor-has been developed to produce a regenerative biological response. Unlike conventional hernia meshes, this regenerative 3D hernia scaffold seems to demonstrate suitable features for a pathogenetical and physiological coherent treatment of the disease. The aim of this manuscript is to cross evidence these features through magnetic resonance imaging (MRI) and histology.
The biological response of ProFlor at three defined post-implantation stages has been evaluated through MRI signal intensity and compared to neighbouring muscles and fat. As additional proof, histology of tissue specimens excised at the same post-implantation periods from porcine models during an experimental attempt were also evaluated.
MRI of newly ingrown tissue in ProFlor demonstrated similar signal intensity of muscles while fat tissue showed remarkably higher values. These data matched with the histology of ProFlor biopsies excised from pigs.
The motile compliance to groin movements of ProFlor appears to induce a probiotic biologic response comparable to a regenerative scaffold, allowing to physiologically resolve the degenerative source of inguinal hernia disease.
•MRI evidence of regenerative effect in 3D scaffold for inguinal hernia repair ProFlor in the short, mid and long term post-implantation.•Enhanced biological response into the 3D inguinal hernia scaffold ProFlor histologically verified in porcine experimental model.•Regenerative biologic response of ProFlor results pathogenetically coherent for the cure of the degenerative damages caused by inguinal hernia disease.
The Eurasian woodcock (Scolopax rusticola) belongs to those bird species that make systematic migratory flights in spring and autumn in search of favorable breeding and wintering areas. These ...specimens arrive in the Mediterranean Area from northeastern European countries during the autumn season. The purpose of this study was to assess whether woodcocks can carry antibiotic resistance genes (ARGs) along their migratory routes. Although the role of migratory birds in the spread of some zoonotic diseases (of viral and bacterial etiology) has been elucidated, the role of these animals in the spread of antibiotic resistance has not yet been clarified. In this study, we analyzed the presence of beta-lactam antibiotic resistance genes. The study was conducted on 69 strains from 60 cloacal swabs belonging to an equal number of animals shot during the 2022–2023 hunting season in Sicily, Italy. An antibiogram was performed on all strains using the microdilution method (MIC) and beta-lactam resistance genes were investigated. The strains tested showed no phenotypic resistance to any of the 13 antibiotics tested; however, four isolates of Enterobacter cloacae and three of Klebsiella oxytoca were found to carry the blaIMP-70, blaVIM-35, blaNDM-5 and blaOXA-1 genes. Our results confirm the importance of monitoring antimicrobial resistance among migratory animals capable of long-distance bacteria spread.
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