Chemical Studies of Multicomponent Kidney Stones Using the Modern Advanced Research Methods.

Defining the kidney stone composition is important for determining a treatment plan, understanding etiology and preventing recurrence of nephrolithiasis, which is considered as a common, civilization disease and a serious worldwide medical problem. The aim of this study was to investigate the morphology and chemical composition of multicomponent kidney stones. The identification methods such as infrared spectroscopy (FTIR), X-ray diffraction (XRD), and electron microscopy with the EDX detector were presented. The studies by the X-ray photoelectron spectroscopy (XPS) were also carried out for better understanding of their chemical structure. The chemical mapping by the FTIR microscopy was performed to show the distribution of individual chemical compounds that constitute the building blocks of kidney stones. The use of modern research methods with a particular emphasis on the spectroscopic methods allowed for a thorough examination of the subject of nephrolithiasis.

The sex impact on the technological and chemical characteristics of meat derived from the Polish native chicken breed.

The aim of the study was to evaluate the birds' sex as well as the caponisation influence on the technological traits of obtained meat, fatty acids profile as well as main chemical compounds distribution. In this study, 40 hens, roosters and green-legged partridge capons each were used (4 replications each). At 24 weeks of age, 2 birds from each replication subgroup were selected and slaughtered. During dissection analysis, pectoral and femoral muscles were sampled. Acidity, colour, tenderness, water holding capacity, drip and cooking loss were determined in the obtained material. The fatty acid profile of the meat was also determined, as well as the distribution of components in the pressed meat samples using FTIR spectroscopy. Statistically significant differences in the colour lightness index (L*) of the breast muscles were recorded, with cockerel meat being the darkest and capon meat the lightest. The greatest natural water loss was recorded in the hens' pectoral muscle; cooking loss was also the greatest in these samples. Roosters showed significantly the lowest content of monounsaturated fatty acids, at the same time the highest proportion of the n3 fatty acids group was found in capons. Irrespective of muscle, meat from roosters showed the highest susceptibility to oxidation (PI index). The variation in the chemical composition of the meat was confirmed by FTIR mapping.

Atypical changes in Candida albicans cells treated with the Venetin-1 complex from earthworm coelomic fluid.

In the present research, the effect of a protein-polysaccharide complex Venetin-1 obtained from the coelomic fluid of Dendrobaena veneta earthworm on Candida albicans cells was characterized. The compound destroyed fungal cells without showing cytotoxicity to human skin fibroblasts, which was demonstrated in earlier studies. Since it had an effect on the fungal cell wall and membrane, this complex was compared with the known antifungal antibiotic fluconazole. Both preparations disturbed the division of yeast cells and resulted in the formation of aggregates and chains of unseparated cells, which was illustrated by staining with fluorochromes. Fluorescent staining of the cell wall with Calcofluor white facilitated comparison of the types of aggregates formed after the action of both substances. The analysis performed with the use of Congo red showed that Venetin-1 exposed deeper layers of the cell wall, whereas no such effect was visible after the use of fluconazole. The FTIR analysis confirmed changes in the mannoprotein layer of the cell wall after the application of the Venetin-1 complex. Staining with Rhodamine 123 and the use of flow cytometry allowed comparison of changes in the mitochondria. Significantly elongated mitochondria were observed after the Venetin-1 application, but not after the application of the classic antibiotic. Phase contrast microscopy revealed vacuole enlargement after the Venetin-1 application. The flow cytometry analysis of C. albicans cells treated with Venetin-1 and fluconazole showed that both substances caused a significant decrease in cell viability.

Determination of the Ni(II) Ions Sorption Mechanism on Dowex PSR2 and Dowex PSR3 Ion Exchangers Based on Spectroscopic Studies.

This paper estimates the suitability of the strongly basic anion exchangers, Dowex PSR2 and Dowex PSR3, as sorbents of nickel ions in aqueous solutions. These actions are aimed at searching for new solutions due to the growing discharge of nickel into wastewaters, primarily due to its addition to steel. The nickel sorption experiments were conducted under static conditions and resulted in the optimization of pH, phase contact time, initial solution concentration, and temperature. The next step was to calculate the kinetic, isothermal, and thermodynamic parameters. Moreover, the ion exchangers were characterized by means of Fourier transform infrared spectroscopy, X-ray photoelectron spectroscopy, and CHN elemental analysis. It was found that the sorption process was most effective at pH 6 after 240 min and at the temperature of 293 K. The values of the thermodynamic parameters revealed that the adsorption was exothermic and spontaneous. The physicochemical analyses combined with the experimental research enabled determination of the sorption mechanism of Ni(II) ions.

Novel Venetin-1 nanoparticle from earthworm coelomic fluid as a promising agent for the treatment of non-small cell lung cancer.

The present research shows the antitumor activity of a protein-polysaccharide complex Venetin-1 obtained from the coelomic fluid of Dendrobaena veneta earthworms against A549 cancer cells. The investigations are a continuation of experiments on the antitumor activity of coelomic fluid obtained from this species. The Venetin-1 nanoparticle was obtained after thermal treatment of the coelomic fluid, separation from coelomocytes, filtration, and lyophilization. The preparation showed a selective effect on cancer cells, whereas normal cells were unaffected. Venetin-1 was effective against the lung cancer cells at doses of 31.3 and 62.5 µg/ml, and the results were imaged using light microscopy and scanning electron microscopy (SEM). The cells died mainly via the apoptosis pathway. Necrotic cells appeared sporadically in the microscopic view. SEM imaging revealed complete destruction of the A549 cells after the incubation with Venetin-1. The atomic force microscopy (AFM) analyses showed changes in the topography, peak force error images, and Young's modulus (elasticity) of the A549 cells after the incubation with Venetin-1. The transmission electron cryomicroscopy (Cryo-TEM) analysis indicated a polymeric nature of the analyzed preparation. The samples of Venetin-1 showed a very homogeneous size profile with the microparticle size of approximately 58.23 nm. A significant decrease in Venetin-1 binding to sphingomyelin was observed. Venetin-1 lost its pore-forming activity or deactivation of the pore-forming activity occurred. This confirms the absence of hemolytic capacity of Venetin-1 towards red blood cells. The conducted analyses show the suitability of the obtained complex for biomedical research. The next step will consist in analyses of the effect of Venetin-1 on the immune system in mice.

Analysis of the Microbiome on the Surface of Corroded Titanium Dental Implants in Patients with Periimplantitis and Diode Laser Irradiation as an Aid in the Implant Prosthetic Treatment: An Ex Vivo Study.

The paper presents the optimization of diode laser irradiation of corroded dental implants in order to reduce the number of microorganisms associated peri-implantitis. The research included the identification of microorganisms on the surface of removed dental implants in patients with peri-implantitis and the assessment of the biocidal effectiveness of the diode laser against these microorganisms. Laser desorption/mass spectrometry (MALDI-TOF MS) was used to identify microorganisms and metagens were examined by next generation sequencing (NGS). Irradiation was performed with a diode laser with a wavelength of λ = 810, operating mode: 25 W/15.000 Hz/10 µs, average = 3.84 W with the number of repetitions t = 2 × 15 s and t = 3 × 15 s. The structure and surface roughness of the implants were analysed before and after laser irradiation by optical profilometry and optical microscopy with confocal fixation. In total, 16 species of Gram-positive bacteria and 23 species of Gram-negative bacteria were identified on the surface of the implants. A total of 25 species of anaerobic bacteria and 12 species with corrosive potential were detected. After diode laser irradiation, the reduction in bacteria on the implants ranged from 88.85% to 100%, and the reduction in fungi from 87.75% to 96.77%. The reduction in microorganisms in the abutment was greater than in the endosseous fixture. The applied laser doses did not damage, but only cleaned the surface of the titanium implants. After 8 years of embedding, the removed titanium implant showed greater roughness than the 25-year-old implant, which was not exposed to direct influence of the oral cavity environment. The use of a diode laser in an optimised irradiation dose safely reduces the number of microorganisms identified on corroded dental implants in patients with peri-implantitis.

The Use of the Diode Laser against the Microbiome on Composites Closing the Screw Access Hall (Sah) in the Reconstruction of Dental Implants: Ex Vivo Studies.

Patients undergoing implant treatment are at risk of peri-implant bone loss, which is most often caused by the adverse effects of microorganisms, but there are few proven procedures for their reduction. The aim of the research was to identify the microorganisms inhabiting the composites used to close the screw access hole (SAH), compare them numerically with those present on the surface of crowns and teeth, and optimize the doses of the diode laser, which will reduce microorganisms and will not deteriorate the roughness of polished composites. Patients were swabbed from the surface of SAH composites, from porcelain and zirconium restorations, and from teeth, and then the number of microorganisms was determined by using a culture technique. Microorganisms were identified by MALDI-TOF MS and NGS sequencing. The effectiveness of diode laser irradiation was achieved by using four variants of exposure. After polishing and laser irradiation, the surface roughness of the composites was studied by using optical profilometry. On the surface of SAH, 106 to 108 microorganisms were identified at 0.4 cm2, including many pathogenic species. Among the materials used for the reconstruction of dental implants, the greatest microbiological contamination was found on the composites used to close the SAH. The diode laser with a wavelength of 810 nm with an average power of 3.84 W, during 60 s and 2 × 30 s, has a biocidal effect and does not significantly change the surface roughness of composites. The best reduction of microorganisms was achieved on a composite polished with a polishing rubber and then with a Sof-Lex™ Pre-Polishing Spiral beige (3M ESPE, Ave. St. Paul., MN, USA). Studies have shown that using the optimal laser dose can help treat periimplantitis. These studies provide important information on the possibility of the effective elimination of microorganisms by using a diode laser in the treatment of peri-implant bone loss.

How the Duration and Mode of Photopolymerization Affect the Mechanical Properties of a Dental Composite Resin.

Composite materials are the most common materials in use in modern dentistry. Over the years, the methods of photopolymerization of composite materials have been improved with the use of various devices, such as quartz tungsten halogen lamps (QTHs), light-emitting diode units (LEDs), plasma-arc lamps and argon-ion lasers. This study aimed to compare the mechanical properties of a composite material, depending on the time and mode of photopolymerization. One hundred and forty rectangular specimens (25 × 2 × 2 mm) and forty-two disc-shaped samples (5 mm diameter and 2 mm thickness) were prepared from shade A2 Boston composite resin. Samples were cured using the following seven photopolymerization protocols: four fast-cure modes (full power for 3, 5, 10, and 20 s), two pulse-cure modes (5 and 10 shots of 1 s exposures at full power), and one step-cure mode (soft start with a progressive cycle lasting 9 s). Specimens were subjected to a flexural strength test, Vickers microhardness test, and FTIR spectroscopy test. A 2-factor ANOVA and post-hoc tests were carried out to assess the differences in the flexural strength parameter between the tested groups of samples before and after aging. A mixed-model ANOVA was carried out to assess the differences in the Vickers microhardness parameter between the tested groups of samples before and after aging. The lowest values of flexural strength (p < 0.001) and Vickers microhardness (p < 0.001) were obtained for the 3 s mode for the pre- and post-aging groups. The FTIR mapping tests showed a much more homogeneous chemical structure of the composite after 20 s of continuous irradiation, compared to the sample irradiated for 5 s in the continuous mode. The mode and cure time affects the mechanical properties of the composite resin. Appropriate selection of the cure mode and time ensures better mechanical properties of composite resin. This suggests that the survival of dental restorations within the oral cavity could be extended by using longer photopolymerization durations.

The Efficacy of a Diode Laser on Titanium Implants for the Reduction of Microorganisms That Cause Periimplantitis.

The paper presents the optimisation of a safe diode laser irradiation process applied to the surface of titanium implants in order to reduce microbial numbers in the treatment of inflammation classified as periimplantitis. The study comprised isolation and identification of microorganisms inhabiting surfaces of dental implants, crowns, teeth and saliva from patients with fully symptomatic periimplantitis. Microorganisms were detected by a culture-dependent method and identified with the use of MALDI-TOF mass spectrometry. The isolated microorganisms were inoculated on the surface of a new implant and then irradiated by a diode laser (wavelength of 810 ± 10 nm) in one, two or three repetitions and biocidal efficacy was assessed. To evaluate impact of laser irradiation on roughness, morphology and structure of the implant surface, optical profilometry, scanning electron microscopy and optical microscopy were used. Examination of the tested surfaces and saliva revealed the presence of Gram-positive and Gram-negative bacteria and one fungal species. In all patients, cultures from the endosseous part of the implant revealed the presence of the pathogenic and pyogenic bacterium Streptococcus constellatus. In 13 out of 20 samples laser-irradiated in duplicate and triplicate, all microorganisms were eliminated. The irradiation used did not cause any changes in the properties of the implant surface.

Application of Modern Research Methods for the Physicochemical Characterization of Ion Exchangers.

Ion exchange technique as the reversible exchange of ions between the substrate and the surrounding medium can be an effective way of removing traces of ion impurities from the waters and wastewaters and obtaining a product of ultrapure quality. Therefore, it can be used in analytical chemistry, hydrometallurgy, purification and separation of metal ions, radioisotopes and organic compounds, and it also finds great application in water treatment and pollution control. In the presented paper, the new trends for ion exchanger characteristics determination and application are presented. Special attention is paid to the ion exchangers with multifunctionality for heavy metal ions removal. They show superior actions such as sorption capacity values with excellent resistance to fouling and the possibility of application in the co-current or modern packed bed counter-current systems, as well as for the condensate polishing or the conventional mixed bed systems in combination with other resins. The results of the paper are expected to help researchers to establish a powerful strategy to find a suitable ion exchanger for heavy metal ions removal from waters and wastewaters. It is important because the best ion exchangers are selected for a specific application during laboratory tests taking into account the composition of the feed solution, pH, type of ion exchangers and then the column breakthrough tests. Therefore, the optical profilometry and the X-ray photoelectron spectroscopy can prove beneficial for this purpose in the case of three different ion exchangers such as Dowex M 4195, Amberlite IRA 743 and Purolite Arsen Xnp.

The Use of a Diode Laser for Removal of Microorganisms from the Surfaces of Zirconia and Porcelain Applied to Superstructure Dental Implants.

The aim of this paper was to study the effectiveness of a diode laser (LD) for removal of microorganisms isolated from porcelain and zirconia crown surfaces used in implantoprosthetics in order to minimize infections around dental implants. In order to optimize biocidal efficacy of the process (at the same time, avoiding increasing the surface roughness during decontamination) the effects of diode laser doses were investigated. The irradiation was performed with a diode laser at the wavelength of λ = 810 nm in three variants with a different number of repetitions (1 × 15 s, 2 × 15 s, 3 × 15 s). The quantitative microbial contamination of the surface of teeth, porcelain and zirconia crowns assessment was made using the culture-dependent method. The identification of microorganisms took place using the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) and next-generation sequencing (NGS) methods. The studies of the surface morphology and roughness were carried out by means of the optical profilometry, scanning electron microscopy (SEM) and optical microscopy with the C1 confocal attachment. The most important conclusion from the research is the fact that the laser operation, regardless of the exposure time, effectively eliminates the microorganisms from the surfaces used for dental implant rebuilding and does not have a destructive effect on the tested material.

Morphological and spectroscopic analysis of snow and glacier algae and their parasitic fungi on different glaciers of Svalbard.

The results show the morphological analyses and spectroscopic studies of snow and glacier algae and their parasitic fungi in Svalbard (High Arctic). Fixed algal cells of two species, Sanguina nivaloides and Ancylonema nordenskioeldii, were imaged using light microscopy, scanning electron microscopy (SEM), and atomic force microscopy (AFM). Fluorescence microscopy using Calcofluor white stain supported the observations of parasitic fungi on the algal cells. Images in brightfield microscopy showed chytrid-like fungi penetrating the cells of both algal species. Parasites were found to colonize the cells of A. nordenskioeldii and hypnozygotes of S. nivaloides, while no fungi infected the cyst stages of S. nivaloides. The autofluorescence analysis revealed the ability of S. nivaloides to glow when excited with different wavelengths, while A. nordenskioeldii did not fluoresce. The hypnozygotes of S. nivaloides emitted brighter fluorescence than the cysts, and the most intense luminosity was observed in the UV range. The Fourier-transform infrared spectroscopy (FTIR) and energy-dispersive X-ray spectroscopy (EDS) spectroscopic analysis showed differences in the chemical composition between samples collected from three different sites. Samples dominated by cyst cells were characterized by the presence of an abundant polysaccharide envelope.

Metabolic, structural, and proteomic changes in Candida albicans cells induced by the protein-carbohydrate fraction of Dendrobaena veneta coelomic fluid.

The isolated protein-polysaccharide fraction (AAF) from the coelomic fluid of Dendrobaena veneta earthworm shows effective activity against Candida albicans yeast. Fungal cells of the clinical strain after incubation with the active fraction were characterized by disturbed cell division and different morphological forms due to the inability to separate the cells from each other. Staining of the cells with acridine orange revealed a change in the pH of the AAF-treated cells. It was observed that, after the AAF treatment, the mitochondrial DNA migrated towards the nuclear DNA, whereupon both merged into a single nuclear structure, which preceded the apoptotic process. Cells with a large nucleus were imaged with the scanning electron cryomicroscopy (Cryo-SEM) technique, while enlarged mitochondria and the degeneration of cell structures were shown by transmission electron microscopy (TEM). The loss of the correct cell shape and cell wall integrity was visualized by both the TEM and SEM techniques. Mass spectrometry and relative quantitative SWATH MS analysis were used to determine the reaction of the C. albicans proteome to the components of the AAF fraction. AAF was observed to influence the expression of mitochondrial and oxidative stress proteins. The oxidative stress in C. albicans cells caused by the action of AAF was demonstrated by fluorescence microscopy, proteomic methods, and XPS spectroscopy. The secondary structure of AAF proteins was characterized by Raman spectroscopy. Analysis of the elemental composition of AAF confirmed the homogeneity of the preparation. The observed action of AAF, which targets not only the cell wall but also the mitochondria, makes the preparation a potential antifungal drug killing the cells of the C. albicans pathogen through apoptosis.

Medical Plant Extract Purification from Cadmium(II) Using Modified Thermoplastic Starch and Ion Exchangers.

Pure compounds extracted and purified from medical plants are crucial for preparation of the herbal products applied in many countries as drugs for the treatment of diseases all over the world. Such products should be free from toxic heavy metals; therefore, their elimination or removal in all steps of production is very important. Hence, the purpose of this paper was purification of an extract obtained from Dendrobium officinale Kimura et Migo and cadmium removal using thermoplastic starch (S1), modified TPS with poly (butylene succinate); 25% of TPS + 75% PBS (S2); 50% of TPS + 50% PLA (S3); and 50% of TPS + 50% PLA with 5% of hemp fibers (S4), as well as ion exchangers of different types, e.g., Lewatit SP112, Purolite S940, Amberlite IRC747, Amberlite IRC748, Amberlite IRC718, Lewatit TP207, Lewatit TP208, and Purolite S930. This extract is used in cancer treatment in traditional Chinese medicine (TCM). Attenuated total reflectance-Fourier transform infrared spectroscopy, thermogravimetric analysis with differential scanning calorimetry, X-ray powder diffraction, gel permeation chromatography, surface analysis, scanning electron microscopy with energy dispersive X-ray spectroscopy, and point of zero charge analysis were used for sorbent and adsorption process characterization, as well as for explanation of the Cd(II) sorption mechanism.

Studies on the Mechanism of Cu(II) Ion Sorption on Purolite S 940 and Purolite S 950.

The aim of the presented research was to investigate the mechanism of sorption of Cu(II) ions on the commercially available Purolite S 940 and Purolite S 950 chelating ion exchangers with the aminophosphonic functional groups. In order to understand better the sorption mechanism, the beads were cut with an ultramicrotome before and after the Cu(II) ion sorption process. The cut beads were examined by scanning electron microscopy (SEM) with an EDX detector. The performed linear profiles of the elemental composition allowed us to examine the depth with which the sorbed metal penetrates into. For further investigations concerning the mechanism of the sorption process, the Fourier transform infrared spectroscopy (FTIR) analysis using the attenuated total reflectance (ATR) technique and the X-ray photoelectron spectroscopy (XPS) methods have been used. The comparison of FTIR and XPS spectra before and after the sorption of Cu(II) ions showed that free electron pairs from nitrogen and oxygen in the aminophosphonic functional groups participate in the process of copper ion sorption. In addition, the microscopic studies suggested that the process of ion exchange between Na(I) ions and sorbed Cu(II) ions takes place on the Purolite S 940 and Purolite S 950. This study concerning the in-depth understanding the of Cu(II) sorption mechanism, using modern analytical tools and research methods could be very useful for its further modifications leading to the improvement of the process efficiency.

Impact of Sinorhizobium meliloti Exopolysaccharide on Adsorption and Aggregation in the Copper(II) Ions/Supporting Electrolyte/Kaolinite System.

To obtain insight into physicochemical interactions between Cu(II) ions, kaolinite, and exopolysaccharide (EPS) synthesized by Sinorhizobium meliloti Rm 1021 soil bacteria, an adsorption, electrokinetic, and aggregation study was performed in the selected systems. The obtained data showed that supporting electrolyte type affects both EPS and Cu(II) ions adsorption. For initial Cu(II) concentration 100 mg/L, 4.36 ± 0.25 mg/g (21.80 ± 1.00%) of the ions were adsorbed in 0.001 M NaCl and 3.76 ± 0.20 mg/g (18.80 ± 1.00%) in 0.001 M CaCl2. The experimental data were best fitted to the Langmuir model as well as pseudo second-order equation. The EPS adsorbed amount on kaolinite was higher in the CaCl2 electrolyte than in NaCl one. For an initial polymer concentration of 100 mg/L, the EPS adsorbed amount was 4.69 ± 0.08 mg/g (23.45 ± 0.40%) in 0.001 M NaCl and 5.26 ± 0.15 mg/g (26.32 ± 0.75%) in 0.001 M CaCl2. In the mixed system, regardless of electrolyte type, exopolysaccharide contributed to immobilization of higher amount of copper(II) ions on the clay mineral. Also, in the samples containing heavy metal ions and exopolysaccharide simultaneously, the aggregation of kaolinite particles was the strongest. The results presented in the paper may be very helpful in soil bioremediation, especially in the development of technologies reducing the mobility of heavy metals in the environment.

Arsenate Adsorption on Fly Ash, Chitosan and Their Composites and Its Relations with Surface, Charge and Pore Properties of the Sorbents.

One of the ways to recycle millions of tons of fly ash and chitin wastes produced yearly is their utilization as low-cost sorbents, mainly for heavy metal cations and organic substances. To improve their sorption efficiency, fly ashes have been thermally activated or modified by chitosan. We aimed to deeply characterize the physicochemical properties of such sorbents to reveal the usefulness of modification procedures and their effect on As(V) adsorption. Using low temperature nitrogen adsorption, scanning electron microscopy, mercury intrusion porosimetry, potentiometric titration and adsorption isotherms of As(V) anions, surface, pore, charge and anion adsorption parameters of fly ash activated at various temperatures, chitosan, and fly ash modified by chitosan were determined. Arsenate adsorption equilibrium (Langmuir model), kinetics (pseudo-second order model) and thermodynamics on the obtained materials were studied. Neither temperature activation nor chitosan modifications of fly ash were necessary and profitable for improving physicochemical properties and As(V) adsorption efficiency of fly ash. Practically, the physicochemical parameters of the sorbents were not related to their anion adsorption parameters.

Morphological and physicochemical diversity of snow algae from Alaska.

Snow algae are photosynthetic microbes growing in thawing snow. They usually show various morphological cell types. The aim of this study was to carry out microscopic and spectroscopic analysis of different forms of cells of snow algae collected on glaciers in Alaska. Four different shapes of algal cells were observed with the use of bright field LM (Light Microscopy), DIC (Differential Interference Contrast), EDF (Extended Depth Focus), fluorescence microscopy, and SEM (Scanning Electron Microscopy). The cells exhibited the strongest autofluorescence after the exposure to 365-nm excitation light, and the intensity differed among the cell types. Zygotes (cysts) showed the most intense fluorescence. Acridine orange staining revealed the acid nature of the algal cells. The use of Congo red and Calcofluor white fluorochromes indicated differences in the structure of polysaccharides in the cell wall in the individual types of algal cells. FTIR (Fourier-Transform Infrared Spectroscopy) analyses showed the presence of polysaccharides not only in the algal cells but also in the fixative solution. The presence of polysaccharides in the extracellular algal fraction was confirmed by X-ray dispersion spectroscopy (EDS), X-ray photoelectron spectroscopy (XPS), and scanning electron microscopy imaging (SEM). The differences observed in the structure of the cell wall of the different forms of red snow algae prompt further analysis of this structure.

Candida albicans cell wall as a target of action for the protein-carbohydrate fraction from coelomic fluid of Dendrobaena veneta.

The protein-polysaccharide fraction (AAF) isolated from the coelomic fluid of the earthworm Dendrobaena veneta destroys C. albicans cells by changing their morphology, disrupting cell division, and leading to cell death. Morphological changes in C. albicans cells induced by treatment with AAF were documented using DIC, SEM, and AFM. Congo Red staining showed that the fungal wall structure was changed after incubation with AAF. The effect on C. albicans cell walls was shown by AFM analysis of the surface roughness of fungal cell walls and changes in the wall thickness were visualized using Cryo-SEM. The FTIR analysis of C. albicans cells incubated with AAF indicated attachment of protein or peptide compounds to the fungal walls. The intact LC-ESI-MS analysis allowed accurate determination of the masses of molecules present in AAF. As shown by the chromatographic study, the fraction does not cross biological membranes. The Cryo-TEM analysis of AAF demonstrated the ability of smaller subunits to combine into larger agglomerates. AAF is thermally stable, which was confirmed by Raman spectroscopy. AAF can be considered as a potential antifungal antibiotic with activity against clinical C. albicans strains.

Antifungal and anticancer effects of a polysaccharide-protein complex from the gut bacterium Raoultella ornithinolytica isolated from the earthworm Dendrobaena veneta.

The polysaccharide-protein complex (PPC) isolated from metabolites of gut bacteria Raoultella ornithinolytica from Dendrobaena veneta earthworms exhibits activity against Candida albicans, in breast ductal carcinoma (line T47D) and in the endometrioid ovarian cancer line (TOV-112D) in vitro. The action against C. albicans was analyzed using light, SEM, TEM, and AFM microscopes. The changes observed indicated two directions of the action of the complex, that is, disturbance of metabolic activity and cell wall damage. The PPC is an adhesion-promoting complex inducing death of C. albicans cells by necrosis. Owing to its significant effect on C. albicans, the complex is a promising source of antifungal compounds. The PPC showed a minimal cytotoxic effect against human skin fibroblasts; however, the cytotoxicity against the T47D line was determined at 20% and 15% against the TOV-112D line. The action of the PPC against the T47D line exerted a cytopathic effect, whereas in the TOV-112D line, it caused a reduction in the cell number. The PPC induced death of tumor cells by apoptosis and necrosis. In view of the negligible cytotoxicity on fibroblasts, the PPC will be subjected to chemical modifications to increase its antitumor activity for prospective medical applications.