Theranostic Approach for the Protein Corona of Polysaccharide Nanoparticles.

Polysaccharide nanoparticles are promising materials in the wide range of disciplines such as medicine, nutrition, food production, agriculture, material science and others. They excel not only in their non-toxicity and biodegradability but also in their easy preparation. As well as inorganic particles, a protein corona (PC) around polysaccharide nanoparticles is formed in biofluids. Moreover, it has been considered that the overall response of the organism to nanoparticles presence depends on the PC. This review summarises scientific publications about the structural chemistry of polysaccharide nanoparticles and their impact on theranostic applications. Three strategies of implementation of the PC in theranostics have been discussed: I) Utilisation of the PC in therapy; II) How the composition of the PC is analysed for specific disease markers; III) How the formed PC can interact with the immune system and enhances the immunomodulation or immunoelimination. Thus, the findings from this review can contribute to improve the design of drug delivery systems. However, it is still necessary to elucidate the mechanisms of nano-bio interactions and discover new connections in nanoscale research.

Development of pH-Responsive Biopolymeric Nanocapsule for Antibacterial Essential Oils.

It is generally believed that antibacterial essential oils have the potential to become one of the alternatives in preventing diarrheal diseases of monogastric animals. The disadvantage is their low efficiency per oral due to easy degradation during digestion in the stomach. This study compares the efficacy of chitosan, alginate-chitosan, guar gum-chitosan, xanthan gum-chitosan and pectin-chitosan nanocapsules to the synthesis of pH-responsive biopolymeric nanocapsule for Thymus vulgaris, Rosmarinus officinalis and Syzygium aromaticum essential oils. Using spectrophotometric approach and gas chromatography, release kinetics were determined in pH 3, 5.6 and 7.4. The growth rates of S. aureus and E. coli, as well as minimal inhibition concentration of essential oils were studied. The average encapsulation efficiency was 60%, and the loading efficiency was 70%. The size of the nanocapsules ranged from 100 nm to 500 nm. Results showed that chitosan-guar gum and chitosan-pectin nanocapsules released 30% of essential oils (EOs) at pH 3 and 80% at pH 7.4 during 3 h. Similar release kinetics were confirmed for thymol, eugenol and α-pinene. Minimal inhibition concentrations of Thymus vulgaris and Syzygium aromaticum essential oils ranged from 0.025 to 0.5%. Findings of this study suggest that the suitable pH-responsive nanocapsule for release, low toxicity and antibacterial activity is based on chitosan-guar gum structure.

Zinc Modified Nanotransporter of Anticancer Drugs for Targeted Therapy: Biophysical Analysis.

Modern anticancer therapy aims to increase the effectiveness of tumor treatment. The aim of this work was to propose a new nanotransporter for targeted delivery of anthracycline antibiotics, which is characterized by its bioavailability, increased uptake of the drug from the bloodstream at the site of the tumor tissue as well as low toxicity to non-target tissue. Chitosan nanoparticles have attracted great attention in the field of drug delivery due to their stability, low toxicity and easy preparation. Deacetylated chitosan skeleton is composed of glucosamine units and has a high density of charged amino groups which allow strong electrostatic interactions with biomolecules, transition metals (Zn, Se) and peptides. We obtained an effective level of chitosan encapsulation, 20%. Electrochemical detection of the bounded Zn2+ ions into the chitosan structure showed a potential shift from -0.99 to -0.93 V. This result proved the formation of a chitosan-zinc complex. The ability of metallothione to quench the 2,2-diphenyl-1-picrylhydrazyl radical in the presence of 50 µM doxorubicin was confirmed by the change of relative absorbance over the range from 50 to 60%.

Development of New Silver Nanoparticles Suitable for Materials with Antimicrobial Properties.

Silver nanoparticles are the most important nanoparticles in connection with the antimicrobial effect. Nowadays, the green synthesis of various types of nanoparticles is rapid, effective and produce less toxic nanoparticles often with specific properties. In our experiment we have developed and described in details various types of silver nanoparticles synthesized chemically or by the green synthesis. Nine different silver nanoparticles were synthesized, three by citrate method at different pHs (8; 9; 10), four using gallic acid at alkaline pHs (10; 11), and two by green synthesis using green tea and coffee extracts, both at pH 9. Characterisation of silver nanoparticles was performed using dynamic light scattering, scanning electron microscopy, and ultraviolet-visible absorption spectroscopy. Silver nanoparticles prepared by green synthesis showed the highest antioxidant activity and also ability for quenching of free radicals. Antibacterial activity of silver nanoparticles was determined on bacterial cultures such as Staphylococcus aureus and Escherichia coli. Silver nanoparticles synthesized using green tea and coffee extracts showed the highest antibacterial activity for both bacterial strains. Minimal inhibition concentration for both strains was found to be 65 µM at each silver nanoparticle synthesized using green synthesis.

A Summary of New Findings on the Biological Effects of Selenium in Selected Animal Species-A Critical Review.

Selenium is an essential trace element important for many physiological processes, especially for the functions of immune and reproductive systems, metabolism of thyroid hormones, as well as antioxidant defense. Selenium deficiency is usually manifested by an increased incidence of retention of placenta, metritis, mastitis, aborts, lowering fertility and increased susceptibility to infections. In calves, lambs and kids, the selenium deficiency demonstrates by WMD (white muscle disease), in foals and donkey foals, it is associated with incidence of WMD and yellow fat disease, and in pigs it causes VESD (vitamin E/selenium deficiency) syndrome. The prevention of these health disorders can be achieved by an adequate selenium supplementation to the diet. The review summarizes the survey of knowledge on selenium, its biological significance in the organism, the impact of its deficiency in mammalian livestock (comparison of ruminants vs. non-ruminants, herbivore vs. omnivore) and possibilities of its peroral administration. The databases employed were as follows: Web of Science, PubMed, MEDLINE and Google Scholar.

Fluorescence Characterization of Gold Modified Liposomes with Antisense N-myc DNA Bound to the Magnetisable Particles with Encapsulated Anticancer Drugs (Doxorubicin, Ellipticine and Etoposide).

Liposome-based drug delivery systems hold great potential for cancer therapy. The aim of this study was to design a nanodevice for targeted anchoring of liposomes (with and without cholesterol) with encapsulated anticancer drugs and antisense N-myc gene oligonucleotide attached to its surface. To meet this main aim, liposomes with encapsulated doxorubicin, ellipticine and etoposide were prepared. They were further characterized by measuring their fluorescence intensity, whereas the encapsulation efficiency was estimated to be 16%. The hybridization process of individual oligonucleotides forming the nanoconstruct was investigated spectrophotometrically and electrochemically. The concentrations of ellipticine, doxorubicin and etoposide attached to the nanoconstruct in gold nanoparticle-modified liposomes were found to be 14, 5 and 2 µg·mL(-1), respectively. The study succeeded in demonstrating that liposomes are suitable for the transport of anticancer drugs and the antisense oligonucleotide, which can block the expression of the N-myc gene.

Fabrication of solid-state nanopores and its perspectives.

Nanofluidics is becoming an extensively developing technique in the field of bioanalytical chemistry. Nanoscale hole embed in an insulating membrane is employed in a vast variety of sensing platforms and applications. Although, biological nanopores have several attractive characteristics, in this paper, we focused on the solid-state nanopores due to their advantages as high stability, possibility of diameter control, and ease of surface functionalizing. A detection method, based on the translocation of analyzed molecules through nanochannels under applied voltage bias and resistive pulse sensing, is well established. Nevertheless, it seems that the new detection methods like measuring of transverse electron tunneling using nanogap electrodes or optical detection can offer significant additional advantages. The aim of this review is not to cite all related articles, but highlight the steps, which in our opinion, meant important progresses in solid-state nanopore analysis.

3D-printed chip for detection of methicillin-resistant Staphylococcus aureus labeled with gold nanoparticles.

Methicillin-resistant Staphylococcus aureus (MRSA) is a dangerous pathogen occurring not only in hospitals but also in foodstuff. Currently, discussions on the issue of the increasing resistance, and timely and rapid diagnostic of resistance strains have become more frequent and sought. Therefore, the aim of this study was to design an effective platform for DNA isolation from different species of microorganisms as well as the amplification of mecA gene that encodes the resistance to ß-lactam antibiotic formation and is contained in MRSA. For this purpose, we fabricated 3D-printed chip that was suitable for bacterial cultivation, DNA isolation, PCR, and detection of amplified gene using gold nanoparticle (AuNP) probes as an indicator of MRSA. Confirmation of the MRSA presence in the samples was based on a specific interaction between mecA gene with the AuNP probes and a colorimetric detection, which utilized the noncross-linking aggregation phenomenon of DNA-functionalized AuNPs. To test the whole system, we analyzed several real refractive indexes, in which two of them were positively scanned to find the presence of mecA gene. The aggregation of AuNP probes were reflected by 75% decrease of absorbance (λ = 530 nm) and change in AuNPs size from 3 ± 0.05 to 4 ± 0.05 nm (n = 5). We provide the one-step identification of mecA gene using the unique platform that employs the rapid, low-cost, and easy-to-use colorimetric method for MRSA detection in various samples.

Preconcentration based on paramagnetic microparticles for the separation of sarcosine using hydrophilic interaction liquid chromatography coupled with coulometric detection.

Sarcosine has been identified as a potential prostate cancer marker. To provide determination of this compound, a number of methods are developing. In this study, we optimized a method for its separation by hydrophilic interaction LC with electrochemical detection (ED). Due to the fact that mobile phases commonly used for this type of separation altered the LODs measured by electrochemical detectors, we applied postcolumn dosing of buffer suitable for ED. The optimized conditions were mobile phase A acetonitrile, mobile phase B water in the ratio A/B 70:30, with postcolumn addition of mobile phase C (200 mM phosphate buffer pH 9). The optimal mixing ratio was A + B/C 1:1 with a flow rate of 0.80 mL/min (0.40 + 0.40 mL/min) and detection potential of 1000 mV. Due to the optimization of the parameters for effective separation, which had to meet the optimal parameters of ED, we reached a good resolution for separation also with a good LOD (100 nM). In addition, we successfully carried out sarcosine analysis bound on our modified paramagnetic microparticles with the ability to preconcentrate sarcosine isolated from artificial urine.

Modulation of induced cytotoxicity of doxorubicin by using apoferritin and liposomal cages.

Doxorubicin is an effective chemotherapeutic drug, however, its toxicity is a significant limitation in therapy. Encapsulation of doxorubicin inside liposomes or ferritin cages decreases cardiotoxicity while maintaining anticancer potency. We synthesized novel apoferritin- and liposome-encapsulated forms of doxorubicin ("Apodox" and "lip-8-dox") and compared its toxicity with doxorubicin and Myocet on prostate cell lines. Three different prostatic cell lines PNT1A, 22Rv1, and LNCaP were chosen. The toxicity of the modified doxorubicin forms was compared to conventional doxorubicin using the MTT assay, real-time cell impedance-based cell growth method (RTCA), and flow cytometry. The efficiency of doxorubicin entrapment was 56% in apoferritin cages and 42% in the liposome carrier. The accuracy of the RTCA system was verified by flow-cytometric analysis of cell viability. The doxorubicin half maximal inhibition concentrations (IC50) were determined as 170.5, 234.0, and 169.0 nM for PNT1A, 22Rv1, and LNCaP, respectively by RTCA. Lip8-dox is less toxic on the non-tumor cell line PNT1A compared to doxorubicin, while still maintaining the toxicity to tumorous cell lines similar to doxorubicin or epirubicin (IC50 = 2076.7 nM for PNT1A vs. 935.3 and 729.0 nM for 22Rv1 and LNCaP). Apodox IC50 was determined as follows: 603.1, 1344.2, and 931.2 nM for PNT1A, 22Rv1, and LNCaP.

Selenium Status of Southern Africa.

Selenium is an essential trace element that exists in inorganic forms (selenite and selenates) and organic forms (selenoamino acids, seleno peptides, and selenoproteins). Selenium is known to aid in the function of the immune system for populations where human immunodeficiency virus (HIV) is endemic, as studies suggest that a lack of selenium is associated with a higher risk of mortality among those with HIV. In a recent study conducted in Zambia, adults had a median plasma selenium concentration of 0.27 µmol/L (IQR 0.14-0.43). Concentrations consistent with deficiency (<0.63 µmol/L) were found in 83% of adults. With these results, it can be clearly seen that selenium levels in Southern Africa should be investigated to ensure the good health of both livestock and humans. The recommended selenium dietary requirement of most domesticated livestock is 0.3 mg Se/kg, and in humans above 19 years, anRDA (recommended daily allowance) of 55 mcg Se/per dayisis recommended, but most of the research findings of Southern African countries have recorded low levels. With research findings showing alarming low levels of selenium in soils, humans, and raw feed materials in Southern Africa, further research will be vital in answering questions on how best to improve the selenium status of Southern African soils and plants for livestock and humans to attain sufficient quantities.

Monitoring of taurine dietary supplementation effect on parameters of Duroc boar ejaculate in summer season.

The aim of this experiment was to find out whether the taurine supplementation in daily ration had an effect on quantity or quality of Duroc boar ejaculate. The experiment duration was from June to August, when it could assumed the possible occurrence of heat stress. For the study was chosen 12 Duroc boars of approximately the same age and condition. The control group of 6 Duroc boars was fed only by basic diet and the experimental group of 6 Duroc boars was fed by the same basic diet with supplementation of 15 g taurine/boar/day. Ejaculate was collected once a week by hand glowed technique. From ejaculate parameters were monitored volume of ejaculate, sperm concentration, total amount of sperm, morphologically abnormal sperm, taurine concentration and GSH/GSSH concentration. From microscopic analysis, results were statistically significant in motility in June and July (P<0.05). In biochemical results, a significant difference (P<0.05) has been found between the experimental groups in the concentrations of taurine as well as GSH/GSSG in ejaculate which indicates the effect of heat stress on boars during the experimental period.

Use of nanotechnology-based nanomaterial as a substitute for antibiotics in monogastric animals.

Nanotechnology has emerged as a promising solution for tackling antibiotic resistance in monogastric animals, providing innovative methods to enhance animal health and well-being. This review explores the novel use of nanotechnology-based nanomaterials as substitutes for antibiotics in monogastric animals. With growing global concerns about antibiotic resistance and the need for sustainable practices in animal husbandry, nanotechnology offers a compelling avenue to address these challenges. The objectives of this review are to find out the potential of nanomaterials in improving animal health while reducing reliance on conventional antibiotics. We examine various forms of nanomaterials and their roles in promoting gut health and also emphasize fresh perspectives brought by integrating nanotechnology into animal healthcare. Additionally, we delve into the mechanisms underlying the antibacterial properties of nanomaterials and their effectiveness in combating microbial resistance. By shedding light on the transformative role of nanotechnology in animal production systems. This review contributes to our understanding of how nanotechnology can provide safer and more sustainable alternatives to antibiotics.

Mycotoxin production in different varieties of Dactylis glomerata L. silage in response to biological and chemical additives.

Silage has been identified as a source of different microbial toxins, that may impair farm animal health and productivity as human health can also be compromised. In this sense, the aim of this study was to determine the impact of silage additives on the concentrations of deoxynivalenol (DON) and zearalenone (ZEN) mycotoxins and, eventually, to evaluate the hygienic quality of orchardgrass (Dactylis glomerata L.) silage based on the concentration of them compared to control silage. This study evaluated the influence of biological and chemical additives used in six different varieties of orchardgrass silage on DON and ZEN mycotoxin contents for the first time. The content of both fusariotoxins (DON and ZEN) in fresh matter and grass silage were below the threshold stipulated by the European Commission. The concentration of DON ranges from ~21.86 to 37.26 ng/kg, ~10.21 to 15 ng/kg, ~20.72 to 29.14 ng/kg; and ZEN range from ~3.42 to 7.87 ng/kg, ~3.85 to 8.62 ng/kg and ~2.15 to 5.08 ng/kg, in control, biological and chemical silages, respectively. In general, the biological additive was more efficient for preventing DON contamination, whereas the chemical additive was more efficient for preventing ZEN contamination in grass silage. In summary, the results obtained in this work demonstrate that biological and chemical additives can inhibit fungal growth and mycotoxin production on Dactylis glomerata L. silage and whose use could prevent animal and human diseases.

Microfluidic tool coupled with electrochemical assay for detection of lactoferrin isolated by antibody-modified paramagnetic beads.

Lactoferrin (LF) is approximately 80 kDa iron-binding protein, which is important part of saliva and other body fluids. Due to its ability to bind metal ions, it has many biologically important functions. In this study, a method for the isolation of LF from a biological sample using robotically prepared antibody-modified paramagnetic particles was developed using robotic pipetting station. The method consisted of the following optimised steps. Protein G was bound on the paramagnetic particles, on which goat antibody (10 µg) was linked. LF was subsequently added to microtitration plate, which had affinity to goat antibody and the interaction lasted for 30 min. We found that the highest signals were obtained using the combination of goat antibody 1:3000, murine antibody 1:5000 and conjugate 1:1500. Horseradish peroxidase reducing 3,3,5,5-tetramethylbenzidine (TMB) was linked to the merged complex. The resulted product of this reaction was subsequently analysed spectrometrically with detection limit (3 S/N) as 5 ng/mL. In addition, we also determined TMB by stopped flow injection analysis with electrochemical detection. The limit of detection (3 S/N) was estimated as 0.1 µg/mL. To compare spectrometric and electrochemical approach for detection of TMB, calibration range of bead-LF-antibodies complex was prepared and was determined using a least-squares correlation with coefficient R² higher than 0.95, indicating a very good agreement of the results obtained.

Selenium Nanoparticles as Potential Antioxidants to Improve Semen Quality in Boars.

Selenium is an essential compound which can influence the fertility of boars by a greater margin. In past decades, research was mainly focused on a bioavailability of various selenium forms and the effect on semen quality. Recently, nanotechnology has expanded the possibilities of selenium supplementation research. Twenty-one Duroc boars (three groups with seven boars each) were included in this experiment with the first group being a control group with no selenium supplementation, and the second group being supplemented with 0.3 mg Se/kg of selenium in inorganic form of Na2SeO3. The third group was supplemented with selenium nanoparticles (100 nm) at the same dose as that of the second group. The experiment lasted for 126 days (three spermatogenesis cycles of boars) and the antioxidant parameters of boar semen were analysed at 42, 84 and 126 days, respectively. The antioxidant parameters (DPPH, FRAP, DMPD, GSH, GSSG) were not influenced by both Se2NO3 and selenium nanoparticle supplementation during this experiment. At the end of the monitored period, significantly higher (p < 0.004) antioxidant readings were observed by using the ABTS method but not the DPPH, DMPD and FRAP methods on the supplemented groups compared to the control. Moreover, selenium-nanoparticle-supplemented groups showed elevated glutathione peroxidase activity in the seminal fluid (p < 0.008). However, the selenium nanoparticle supplementation has not shown an improving effect on sperm quality. This could be considered as a safe alternative to inorganic selenium as well as having a potential to enhance the antioxidant properties of the semen of boars.

Toxicological effects of nanoselenium in animals.

The productivity and sustainability of livestock production systems are heavily influenced by animal nutrition. To maintain homeostatic balance in the body of the animal at different phases of life, the percentage of organically active minerals in livestock feed must be optimized. Selenium (Se) is a crucial trace mineral that is required for the maintenance of many functions of the body. Se nanoparticles (SeNPs) attracted considerable interest from researchers for a variety of applications a decade ago, owing to their extraordinary properties. SeNPs offer significant advantages over larger-sized materials, by having a comparatively wider surface area, increased surface energy, and high volume. Despite its benefits, SeNP also has toxic effects, therefore safety concerns must be taken for a successful application. The toxicological effects of SeNPs in animals are characterized by weight loss, and increased mortality rate. A safe-by-strategy to certify animal, human and environmental safety will contribute to an early diagnosis of all risks associated with SeNPs. This review is aimed at describing the beneficial uses and potential toxicity of SeNPs in various animals. It will also serve as a summary of different levels of SeNPs which should be added in the feed of animals for better performance.

Importance of Zinc Nanoparticles for the Intestinal Microbiome of Weaned Piglets.

The scientific community is closely monitoring the replacement of antibiotics with doses of ZnO in weaned piglets. Since 2022, the use of zinc in medical doses has been banned in the European Union. Therefore, pig farmers are looking for other solutions. Some studies have suggested that zinc nanoparticles might replace ZnO for the prevention of diarrhea in weaning piglets. Like ZnO, zinc nanoparticles are effective against pathogenic microorganisms, e.g., Enterobacteriaceae family in vitro and in vivo. However, the effect on probiotic Lactobacillaceae appears to differ for ZnO and zinc nanoparticles. While ZnO increases their numbers, zinc nanoparticles act in the opposite way. These phenomena have been also confirmed by in vitro studies that reported a strong antimicrobial effect of zinc nanoparticles against Lactobacillales order. Contradictory evidence makes this topic still controversial, however. In addition, zinc nanoparticles vary in their morphology and properties based on the method of their synthesis. This makes it difficult to understand the effect of zinc nanoparticles on the intestinal microbiome. This review is aimed at clarifying many circumstances that may affect the action of nanoparticles on the weaning piglets' microbiome, including a comprehensive overview of the zinc nanoparticles in vitro effects on bacterial species occurring in the digestive tract of weaned piglets.

Is a new generation of mycotoxin clay adsorbents safe in a pig's diet?

BACKGROUND: Bentonites, as a clay mineral, serve in pig farms as adsorbents of toxic substances. They are mainly used to reduce the negative impact of mycotoxins to maintain the performance and health status of animals. The new genotypes of pigs are highly sensitive to a range of antinutrients, including mycotoxins. Currently, attention is focused on more effective adsorbents of mycotoxins with a higher adsorption capacity. Such materials are in great demand among feed manufacturers. However, there is a concern that these new materials may also adsorb too many essential nutrients and decrease animal performance. The aim of the experiment was to evaluate the effect of the new generation of purified bentonites on the efficiency and health status of the pigs. RESULTS: Forty-eight slaughtered pigs with an average weight of 31.2 ± 2.6 kg were included in the experiment. The pigs were divided into two groups (2 × 24 pigs). Pigs were slaughtered at an average weight of 66.3 ± 5.2. The first group had a diet without clay (control-C). The second group (treatment-T) was fed a diet with a clay additive (purified bentonite) of 1.5 kg/t. Animals were fed the experimental diet for 35 days. In group T, a higher daily weight gain (by 4.8%) and feed intake (by 2.9%) was observed while the feed conversion decreased by 1.9%. There were no significant differences between the groups of pigs during observation in the evaluation of hematological, biochemical parameters of the blood. Morpho-pathological analysis of the jejunum showed similar signs of moderate lymphoplasmacytic infiltrate in the mucosa in the groups C and T, contained similar number of goblet cells. CONCLUSION: Taken together, the addition of the new generation of bentonite clays did not negatively influence the health status and the performance of pigs.

Effect of Lactic Fermentation and Cooking on Nutrient and Mineral Digestibility of Peas.

Peas are prospectively beneficial legumes in the human diet, and especially in a vegan and vegetarian diet, due to their high content of proteins and starch. Their frequent lack of appeal in human nutrition can be caused by their bloating effect and the content of some antinutritional compounds inhibiting the absorption of important nutrients. This study brings a comprehensive comparison of the nutrient content of pea flour after cooking and lactic fermentation before and after digestion in vitro. As a control sample, raw pea flour was used (sample 1). Raw pea flour was cooked for 10 min (sample 2) and 120 min (sample 3) at 100°C or it was fermented by Lactobacillus plantarum (sample 4) and cooked for 10 min at 100°C (sample 5). The samples were analyzed for protein and amino acids content, maltose, glucose, raffinose, total polyphenols, phytic acid, phytase, and mineral composition (P, Mg, Mn, Fe, Cu, Zn) before and after in vitro digestion. The results showed a significant (p < 0.05) increase in the protein digestibility of samples 3, 4 and 5. In the fermented samples were observed a higher concentration of Cys, Met, and Gln when compared to non-fermented samples. The fermentation of pea flour resulted in a significant (p < 0.05) decrease in glucose, maltose, and raffinose content. Cooking of pea flour for 10 and 120 min, but not fermenting, significantly (p < 0.05) decreased the polyphenols content. Cooking and fermentation together did not affect phytic acid concentration and phytase activity. Mg, Mn, Fe, Cu and, Zn concentration in pea flour was significantly (p < 0.05) decreased by cooking. On the other hand, fermentation significantly (p<0.05) improved the bioaccessibility of Mn and Fe. These findings suggest that lactic fermentation of pea flour is a promising culinary preparation that can improve the digestibility of peas.