Uklanjanje slobodnih radikala, postizanje redoks ravnoteže i cijeljenje rana pomoću bioaktivnih peptida dobivenih hidrolizom kolagena iz kože bijelog glavaša (Hypophthalmichthys molitrix) potpomognutom proteinazom K

Research background. Various protocols for enzymatic hydrolysis of fish by-products are increasingly tested to ensure value-added products with functional and biological properties important for food, cosmetic and medical applications. In addition, they attempt to minimize waste from industrial processing and environmental requirements. This study aims to establish an efficient protocol based on two-step enzymatic hydrolysis of freshwater fish skin and to evaluate the effect of resulting bioactive peptides on free radical scavenging, redox balance and regulation of fibroblast proliferation and migration. Experimental approach. Pepsin-soluble collagen extracted from silver carp (Hypophthalmichthys molitrix) skin was hydrolyzed by proteinase K at specific sites under controlled conditions. The molecular mass of ultrafiltration permeate was determined by gradient electrophoresis and gel filtration chromatography. The biological activity of intermediate and small size bioactive peptides was evaluated in experimental models in vitro mimicking oxidative stress and skin wound conditions. Results and conclusions. Extracted fish collagen was hydrolysed using proteinase K, the most efficient enzyme for the cleavage of the primary structure of the molecule, as previously found in silico. Established optimal conditions increased the enzyme specificity and the process yield. Bioactive peptides exerted significantly higher scavenging activity on free stable radicals and hydroxyl radicals often found in vivo, compared to fish collagen. They stimulated fibroblast metabolism in a dose-dependent manner and up-regulated cell migration in a scratch wound model. Pretreatment of fibroblasts with induced oxidative stress using optimal concentrations of fish peptides prevented the increase of reactive oxygen species production. In conclusion, bioactive peptides from carp skin demonstrated valuable properties of maintaining redox balance and skin wound healing process improvement, which indicated further potential applications in the development of pharmaceutical and nutraceutical formulations. Novelty and scientific contribution. In this study the enzymatic hydrolysis was applied to isolated protein, in contrast to previous studies using waste tissue with variable composition. Recovered bioactive peptides acted not only as antioxidant agents, but also as regulators of oxidative stress and wound healing processes in skin cell models. Their nutritional and cosmetic application is recommended in novel formulations fighting skin ageing phenomena.Pozadina istraživanja. U posljednje se vrijeme sve više ispituju postupci enzimske hidrolize nusproizvoda obrade ribe radi dobivanja proizvoda obogaćenih funkcionalnim i biološkim svojstvima važnim u proizvodnji hrane, kozmetike i medicinskih preparata. Osim toga, na taj se način smanjuju količina otpada industrijske proizvodnje i pritisak na okoliš. Svrha je ovoga rada bila osmisliti učinkoviti postupak dvostupanjske enzimske hidrolize kože slatkovodne ribe i ispitati učinak dobivenih bioaktivnih peptida na uklanjanje slobodnih radikala, redoks ravnotežu, te proliferaciju i migraciju fibroblasta. Eksperimentalni pristup. Kolagen razgradljiv pomoću pepsina izoliran je iz kože bijelog glavaša (Hypoph¬thalmichthys molitrix) i hidroliziran u kontroliranim uvjetima pomoću proteinaze K. Molekulska masa permeata dobivenog ultrafiltracijom određena je pomoću elektroforeze s gradijentom i gel-filtracijske kromatografije. Biološka aktivnost peptida srednje i male veličine ispitana je in vitro simulacijom oksidacijskog stresa i oštećenja kože. Rezultati i zaključci. Riblji kolagen je hidroliziran pomoću proteinaze K, najučinkovitijeg enzima koji cijepa primarnu strukturu molekule, što je potvrđeno prethodnim ispitivanjem in silico. Pri optimalnim uvjetima povećali su se specifičnost enzima i prinos reakcije. U usporedbi s riblijm kolagenom, bioaktivni su peptidi imali bitno veću sposobnost uklanjanja slobodnih i hidroksilnih radikala, često prisutnih in vivo. Ovisno o koncentraciji, stimulirali su metabolizam fibroblasta i migraciju stanica pri cijeljenju modelne rane. Prethodna obrada fibroblasta optimalnim koncentracijama ribljih peptida spriječila je porast sinteze reaktivnih spojeva kisika pri induciranju oksidacijskog stresa. Možemo zaključiti da bioaktivni peptidi kože bijelog glavaša imaju važna svojstva, kao što su održavanje redoks ravnoteže i pospješivanje cijeljenja rana, što upućuje na njihovu moguću primjenu u razvoju farmaceutika i nutraceutika. Novina i znanstveni doprinos. U ovom je radu provedena enzimska hidroliza izoliranog proteina, za razliku od dosadašnjih istraživanja u kojima je korišten otpad različitog sastava. Dobiveni bioaktivni peptidi nisu djelovali samo kao antioksidacijski agensi, već i kao regulatori oksidacijskog stresa i cijeljenja kože u staničnom modelu. Zbog svojih svojstava mogu se preporučiti za primjenu u proizvodnji kozmetičkih pripravaka koji suzbijaju starenje kože

Bio-and hemo-compatible silk fibroin pegylated nanocarriers for 5-fluorouracil chemotherapy in colorectal cancer:In vitro studies

5-fluorouracil (5-FU) remains the gold standard of treatment for colorectal cancer, but its poor bioavailability and high systemic toxicity highlight the urgent need for the development of novel delivery strategies to increase the efficacy of 5-FU treatment. The present study is aimed to design and validate a PEGylated Silk Fibroin Nanocarrier (SF/PEG nanoparticles (NPs)) as an efficient 5-FU delivery system for potential intravenous administration. Using the human adenocarcinoma HT–29 cell line as an in vitro model for colorectal cancer, the cytotoxicity screening of the SF/PEG NPs showed that pristine nanocarriers were highly biocompatible, while the addition of 5-FU triggers a dramatic reduction in tumor cell viability, proliferation potential and mitochondrial integrity as well as a significant increase in nitric oxide production. Despite their high in vitro cytotoxicity, the 5-FU SF/PEG NPs were found hemocompatible as no impact on red blood cells hemolysis or the phagocytic activity of the granulocytes was observed. Exposure of HT–29 tumor cells and blood samples to 5-FU SF/PEG NPs augmented the tumor necrosis factor-α levels. Moreover, 5-FU SF/PEG NPs showed an impact on tumor cell migration and invasive potential as both of these processes were inhibited by the NP treatment

Candida Infections in Severely Burned Patients: 1 Year Retrospective Study

Infections represent the most common complication occurring during the evolution of the severely burned patient, hence requiring closer study and targeted result analysis. The fungal infections are one of the most aggressive types of existing infections, their opportunistic character enabling them to cause invasive infections, ultimately leading to a higher morbidity and a higher rate of mortality. The present study focuses on the presence of Candida spp. in 19 out of a total of 70 patients admitted to the Critical Care Burn Unit in the Clinical Emergency Hospital Bucharest, between 01.01.2019-31.12.2019. No other fungal species, besides Candida spp., were identified in this patient lot. The aim of this study was to analyze the risk factors and the dynamics of the biological parameters of the patients presenting Candida spp. infections, in order to determine how these contribute to the prognostic and final outcome of these patients. We can conclude that a precise diagnostic and prompt treatment can make a significant difference in the outcome of severely burnt patients presenting with a fungal infection

Reducing the environmental impact of surgery on a global scale: systematic review and co-prioritization with healthcare workers in 132 countries

Abstract Background Healthcare cannot achieve net-zero carbon without addressing operating theatres. The aim of this study was to prioritize feasible interventions to reduce the environmental impact of operating theatres. Methods This study adopted a four-phase Delphi consensus co-prioritization methodology. In phase 1, a systematic review of published interventions and global consultation of perioperative healthcare professionals were used to longlist interventions. In phase 2, iterative thematic analysis consolidated comparable interventions into a shortlist. In phase 3, the shortlist was co-prioritized based on patient and clinician views on acceptability, feasibility, and safety. In phase 4, ranked lists of interventions were presented by their relevance to high-income countries and low–middle-income countries. Results In phase 1, 43 interventions were identified, which had low uptake in practice according to 3042 professionals globally. In phase 2, a shortlist of 15 intervention domains was generated. In phase 3, interventions were deemed acceptable for more than 90 per cent of patients except for reducing general anaesthesia (84 per cent) and re-sterilization of ‘single-use’ consumables (86 per cent). In phase 4, the top three shortlisted interventions for high-income countries were: introducing recycling; reducing use of anaesthetic gases; and appropriate clinical waste processing. In phase 4, the top three shortlisted interventions for low–middle-income countries were: introducing reusable surgical devices; reducing use of consumables; and reducing the use of general anaesthesia. Conclusion This is a step toward environmentally sustainable operating environments with actionable interventions applicable to both high– and low–middle–income countries

New Poly(N-isopropylacrylamide-butylacrylate) Copolymer Biointerfaces and Their Characteristic Influence on Cell Behavior In Vitro

Designing and obtaining new synthetic smart biointerfaces with specific and controlled characteristics relevant for applications in biomedical and bioengineering domains represents one of the main challenges in these fields. In this work, Matrix-Assisted Pulsed Laser Evaporation (MAPLE) is used to obtain synthetic biointerfaces of poly(N-isopropyl acrylamide-butyl acrylate) p(NIPAM-BA) copolymer with different characteristics (i.e., roughness, porosity, wettability), and their effect on normal HEK 293 T and murine melanoma B16-F1 cells is studied. For this, the influence of various solvents (chloroform, dimethylsulfoxide, water) and fluence variation (250–450 mJ/cm2) on the morphological, roughness, wettability, and physico–chemical characteristics of the coatings are evaluated by atomic force microscopy, scanning electron microscopy, contact angle measurements, Fourier-transform-IR spectroscopy, and X-ray photoelectron spectroscopy. Coatings obtained by the spin coating method are used for reference. No significant alteration in the chemistry of the surfaces is observed for the coatings obtained by both methods. All p(NIPAM-BA) coatings show hydrophilic character, with the exception of those obtained with chloroform at 250 mJ/cm2. The surface morphology is shown to depend on both solvent type and laser fluence and it ranges from smooth surfaces to rough and porous ones. Physico–chemical and biological analysis reveal that the MAPLE deposition method with fluences of 350–450 mJ/cm2 when using DMSO solvent is more appropriate for bioengineering applications due to the surface characteristics (i.e., pore presence) and to the good compatibility with normal cells and cytotoxicity against melanoma cells

Mitigation of Cellular and Bacterial Adhesion on Laser Modified Poly (2-Methacryloyloxyethyl Phosphorylcholine)/Polydimethylsiloxane Surface

Nowadays, using polymers with specific characteristics to coat the surface of a device to prevent undesired biological responses can represent an optimal strategy for developing new and more efficient implants for biomedical applications. Among them, zwitterionic phosphorylcholine-based polymers are of interest due to their properties to resist cell and bacterial adhesion. In this work, the Matrix-Assisted Laser Evaporation (MAPLE) technique was investigated as a new approach for functionalising Polydimethylsiloxane (PDMS) surfaces with zwitterionic poly(2-Methacryloyloxyethyl-Phosphorylcholine) (pMPC) polymer. Evaluation of the physical–chemical properties of the new coatings revealed that the technique proposed has the advantage of achieving uniform and homogeneous stable moderate hydrophilic pMPC thin layers onto hydrophobic PDMS without any pre-treatment, therefore avoiding the major disadvantage of hydrophobicity recovery. The capacity of modified PDMS surfaces to reduce bacterial adhesion and biofilm formation was tested for Gram-positive bacteria, Staphylococcus aureus (S. aureus), and Gram-negative bacteria, Escherichia coli (E. coli). Cell adhesion, proliferation and morphology of human THP-1 differentiated macrophages and human normal CCD-1070Sk fibroblasts on the different surfaces were also assessed. Biological in vitro investigation revealed a significantly reduced adherence on PDMS–pMPC of both E. coli (from 29 × 10 6 to 3 × 102 CFU/mL) and S. aureus (from 29 × 106 to 3 × 102 CFU/mL) bacterial strains. Additionally, coated surfaces induced a significant inhibition of biofilm formation, an effect observed mainly for E. coli. Moreover, the pMPC coatings improved the capacity of PDMS to reduce the adhesion and proliferation of human macrophages by 50% and of human fibroblast by 40% compared to unmodified scaffold, circumventing undesired cell responses such as inflammation and fibrosis. All these highlighted the potential for the new PDMS–pMPC interfaces obtained by MAPLE to be used in the biomedical field to design new PDMS-based implants exhibiting long-term hydrophilic profile stability and better mitigating foreign body response and microbial infection

Microbiological, Health and Comfort Aspects of Indoor Air Quality in a Romanian Historical Wooden Church

Monitoring the indoor microclimate in old buildings of cultural heritage and significance is a practice of great importance because of the importance of their identity for local communities and national consciousness. Most aged heritage buildings, especially those made of wood, develop an indoor microclimate conducive to the development of microorganisms. This study aims to analyze one wooden church dating back to the 1710s in Romania from the microclimatic perspective, i.e., temperature and relative humidity and the fungal load of the air and surfaces. One further aim was to determine if the internal microclimate of the monument is favorable for the health of parishioners and visitors, as well as for the integrity of the church itself. The research methodology involved monitoring of the microclimate for a period of nine weeks (November 2020–January 2021) and evaluating the fungal load in indoor air as well as on the surfaces. The results show a very high contamination of air and surfaces (>2000 CFU/m3). In terms of fungal contamination, Aspergillus spp. (two different species), Alternaria spp., Cladosporium spp., Mucor spp., Penicillium spp. (two different species) and Trichopyton spp. were the genera of fungi identified in the indoor wooden church air and Aspergillus spp., Cladosporium spp., Penicillium spp. (two different species) and Botrytis spp. on the surfaces (church walls and iconostasis). The results obtained reveal that the internal microclimate not only imposes a potential risk factor for the parishioners and visitors, but also for the preservation of the wooden church as a historical monument, which is facing a crisis of biodeterioration of its artwork

Surgeons’ practice and preferences for the anal fissure treatment: results from an international survey

The best nonoperative or operative anal fissure (AF) treatment is not yet established, and several options have been proposed. Aim is to report the surgeons' practice for the AF treatment. Thirty-four multiple-choice questions were developed. Seven questions were about to participants' demographics and, 27 questions about their clinical practice. Based on the specialty (general surgeon and colorectal surgeon), obtained data were divided and compared between two groups. Five-hundred surgeons were included (321 general and 179 colorectal surgeons). For both groups, duration of symptoms for at least 6 weeks is the most important factor for AF diagnosis (30.6%). Type of AF (acute vs chronic) is the most important factor which guide the therapeutic plan (44.4%). The first treatment of choice for acute AF is ointment application for both groups (59.6%). For the treatment of chronic AF, this data is confirmed by colorectal surgeons (57%), but not by the general surgeons who prefer the lateral internal sphincterotomy (LIS) (31.8%) (p = 0.0001). Botulin toxin injection is most performed by colorectal surgeons (58.7%) in comparison to general surgeons (20.9%) (p = 0.0001). Anal flap is mostly performed by colorectal surgeons (37.4%) in comparison to general surgeons (28.3%) (p = 0.0001). Fissurectomy alone is statistically significantly most performed by general surgeons in comparison to colorectal surgeons (57.9% and 43.6%, respectively) (p = 0.0020). This analysis provides useful information about the clinical practice for the management of a debated topic such as AF treatment. Shared guidelines and consensus especially focused on operative management are required to standardize the treatment and to improve postoperative results