A modified CTAB method for the extraction of high-quality RNA from mono-and dicotyledonous plants rich in secondary metabolites.

BACKGROUND: High-quality RNA extraction from woody plants is difficult because of the presence of polysaccharides and polyphenolics that bind or co-precipitate with the RNA. The CTAB (cetyl trimethylammonium bromide) based method is widely used for the isolation of nucleic acids from polysaccharide-rich plants. Despite the widespread use of the CTAB method, it is necessary to adapt it to particular plant species, tissues and organs. Here we described a simple and generalized method for RNA isolation from mature leaf tissues of several economically important woody (17) and herbaceous plants (2) rich in secondary metabolites. High yields were achieved from small amount (up to 50 mg) of plant material. Two main modifications were applied to the basic protocol: an increase in ß-mercaptoethanol concentration (to 10%v/v) and the use of an effective DNase treatment. As opposed to similar studies, we tried to describe a more detailed protocol for isolating RNA, including the exact quantity and concentration of the reagents were used. RESULTS: Our modified CTAB method is proved to be efficient in extracting the total RNA from a broad range of woody and herbaceous species. The RNA yield was ranged from 2.37 to 91.33 µg/µl. The A260:A280 and A260:A230 absorbance ratios were measured from 1.77 to 2.13 and from 1.81 to 2.22. The RIN value (RNA Integrity Number) of the samples fell between 7.1 and 8.1, which indicated that a small degree of RNA degradation occurred during extraction. The presence of a single peak in the melt curve analyses and low standard errors of the Ct values of replicated measurements indicated the specificity of the primers to bind to the cDNA. CONCLUSIONS: Our RNA isolation method, with fine-tuned and detailed instructions, can produce high quality RNA from a small amount of starting plant material that is suitable for use in downstream transcriptional analyses. The use of an increased concentration of the reducing agent ß-mercaptoethanol in the extraction buffer, as well as the application of DNaseI-treatment resulted in a method suitable for a wide range of plants without the need of further optimalization, especially in Rhus typhina (Staghorn sumac), for which molecular-genetic studies have not yet been sufficiently explored.

Neurovascular Imaging with Ultra-High-Resolution Photon-Counting CT: Preliminary Findings on Image Quality Evaluation.

BACKGROUND AND PURPOSE: The first-generation photon-counting detector (PCD) CT was recently introduced into clinical practice and represents a promising innovation in high-resolution CT imaging. The purpose of this study was to assess the image quality of ultra-high-resolution (UHR) PCD-CT compared with energy-integrating detector (EID)-CT, and to explore different reconstruction kernel sharpness levels for the evaluation of intracranial aneurysms. MATERIALS AND METHODS: Ten patients with intracranial saccular aneurysms, who had previously undergone conventional EID-CT, were prospectively enrolled. CT angiograms were acquired on a clinical dual-source PCD-CT in UHR mode, and reconstructed with four vascular kernels (Bv36, Bv40, Bv44, Bv48). Quantitative and qualitative image quality parameters of the intracranial arteries were evaluated. For the quantitative analysis (image noise, SNR, CNR), regions of interest were manually placed at standard anatomical intracranial and extracranial locations by one author. In addition, vessel border sharpness was evaluated quantitatively. For the qualitative analysis, three blinded neuroradiologists rated PCD-CT and EID-CT image quality for the evaluation of the intracranial vessels (i.e., the aneurysms and nine standard vascular branching locations) on a 5-point Likert-type scale. Additionally, readers independently selected their preferred kernel among the four kernels evaluated on PCD-CT. RESULTS: In terms of quantitative image quality, Bv48, the sharpest kernel, yielded increased image noise, and decreased SNR and CNR parameters compared to Bv36, the smoothest kernel. Compared to EID-CT, the Bv48 kernel offered better quantitative image quality for the evaluation of small intracranial vessels (p < .001). Image quality ratings of the Bv48 were superior to those of the EIDCT, and not significantly different from ratings of the B44 reconstruction kernel. When comparing side-by-side all four PCD-CT reconstruction kernels, readers selected the B48 kernel as the best to visualize the aneurysms in 80% of cases. CONCLUSIONS: UHR PCD-CT provides improved image quality for neurovascular imaging. Although the less sharp kernels provided superior SNR and CNR, the sharpest kernels delivered the best subjective image quality on PCD-CT for the evaluation of intracranial aneurysms.CNR ＝ Contrast-to-Noise Ratio; EID-CT ＝ Energy-Integrating Detector CT; PCD-CT ＝ Photon-Counting Detector CT; QIR ＝ Quantum Iterative Reconstruction; UHR ＝ Ultra-High-Resolution.

Iodine quantification of renal lesions: Preliminary results using spectral-based material extraction on photon-counting CT.

Objectives: To assess the range of quantitative iodine values in renal cysts (RC) (with a few renal neoplasms [RNs] as a comparison) to develop an expected range of values for RC that can be used in future studies for their differentiation. Material and Methods: Consecutive patients (n = 140) with renal lesions who had undergone abdominal examination on a clinical photon-counting computed tomography (PCCT) were retrospectively included. Automated iodine quantification maps were reconstructed, and region of interest (ROI) measurements of iodine concentration (IC) (mg/cm3) were performed on whole renal lesions. In addition, for heterogeneous lesions, a secondary ROI was placed on the area most suspicious for malignancy. The discriminatory values of minimum, maximum, mean, and standard deviation for IC were compared using simple logistic regression and receiver operating characteristic curves (area under the curve [AUC]). Results: A total of 259 renal lesions (243 RC and 16 RN) were analyzed. There were significant differences between RC and RN for all IC measures with the best-performing metrics being mean and maximum IC of the entire lesion ROI (AUC 0.912 and 0.917, respectively) but also mean and minimum IC of the most suspicious area in heterogeneous lesions (AUC 0.983 and 0.992, respectively). Most RC fell within a range of low measured iodine values although a few had higher values. Conclusion: Automated iodine quantification maps reconstructed from clinical PCCT have a high diagnostic ability to differentiate RCs and neoplasms. The data from this pilot study can be used to help establish quantitative values for clinical differentiation of renal lesions.

Apis mellifera filamentous virus from a honey bee gut microbiome survey in Hungary.

In Hungary, as part of a nationwide, climatically balanced survey for a next-generation sequencing-based study of the honey bee (Apis mellifera) gut microbiome, repeated sampling was carried out during the honey production season (March and May 2019). Among other findings, the presence of Apis mellifera filamentous virus (AmFV) was detected in all samples, some at very high levels. AmFV-derived reads were more abundant in the March samples than in the May samples. In March, a higher abundance of AmFV-originated reads was identified in samples collected from warmer areas compared to those collected from cooler areas. A lower proportion of AmFV-derived reads were identified in samples collected in March from the wetter areas than those collected from the drier areas. AmFV-read abundance in samples collected in May showed no significant differences between groups based on either environmental temperature or precipitation. The AmFV abundance correlated negatively with Bartonella apihabitans, Bartonella choladocola, and positively with Frischella perrara, Gilliamella apicola, Gilliamella sp. ESL0443, Lactobacillus apis, Lactobacillus kullabergensis, Lactobacillus sp. IBH004. De novo metagenome assembly of four samples resulted in almost the complete AmFV genome. According to phylogenetic analysis based on DNA polymerase, the Hungarian strains are closest to the strain CH-05 isolated in Switzerland.

Optimisation of virtual monoenergetic reconstructions for the diagnosis of pulmonary embolism using photon-counting detector computed tomography angiography.

Purpose: Computed tomography (CT) pulmonary angiography is considered the gold standard for pulmonary embolism (PE) diagnosis, relying on the discrimination between contrast and embolus. Photon-counting detector CT (PCD-CT) generates monoenergetic reconstructions through energy-resolved detection. Virtual monoenergetic images (VMI) at low keV can be used to improve pulmonary artery opacification. While studies have assessed VMI for PE diagnosis on dual-energy CT (DECT), there is a lack of literature on optimal settings for PCD-CT-PE reconstructions, warranting further investigation. Material and methods: Twenty-five sequential patients who underwent PCD-CT pulmonary angiography for suspicion of acute PE were retrospectively included in this study. Quantitative metrics including signal-to-noise ratio (SNR) and contrast-to-noise (CNR) ratio were calculated for 4 VMI values (40, 60, 80, and 100 keV). Qualitative measures of diagnostic quality were obtained for proximal to distal pulmonary artery branches by 2 cardiothoracic radiologists using a 5-point modified Likert scale. Results: SNR and CNR were highest for the 40 keV VMI (49.3 ± 22.2 and 48.2 ± 22.1, respectively) and were inversely related to monoenergetic keV. Qualitatively, 40 and 60 keV both exhibited excellent diagnostic quality (mean main pulmonary artery: 5.0 ± 0 and 5.0 ± 0; subsegmental pulmonary arteries 4.9 ± 0.1 and 4.9 ± 0.1, respectively) while distal segments at high (80-100) keVs had worse quality. Conclusions: 40 keV was the best individual VMI for the detection of pulmonary embolism by quantitative metrics. Qualitatively, 40-60 keV reconstructions may be used without a significant decrease in subjective quality. VMIs at higher keV lead to reduced opacification of the distal pulmonary arteries, resulting in decreased image quality.

Ixodes ricinus tick bacteriome alterations based on a climatically representative survey in Hungary.

IMPORTANCE: Climate-sensitive disease vectors, such as ticks, respond to the environment with changes in their microbiome. These changes can affect the emergence or re-emergence of various vector-borne pathogens, such as the causative agent of Lyme borreliosis (LB) or tick-borne encephalitis. This aspect is particularly emphasized in light of climate change. The climatically representative assessment of microbiome differences in various developmental stages of the most common Central European tick species, Ixodes ricinus, deepens our understanding of the potential climatic factors behind microbial relative abundance and interaction changes. This knowledge can support the development of novel disease vector control strategies.

First Molecular Identification of Calicophoron daubneyi (Dinnik, 1962) and Paramphistomum leydeni (Nasmark, 1937) in Wild Ruminants from Romania.

Rumen flukes are geographically widespread trematodes affecting wild and domestic ruminants. The juvenile forms, which are found in the small intestine, are more pathogenic compared to the adults. Severe diarrhoea and weight loss are the major clinical signs, and the disease might be fatal in severely infested individuals. In the last decade, paramphistomosis has been described as an emerging parasitic disease in Europe. This study aimed to identify the rumen fluke species in wild ruminants from western Romania. Fifty-two pre-stomachs obtained from roe deer (Capreolus capreolus) that were hunted on 14 hunting grounds from Timiș and Arad counties were examined for the presence of paramphistomes. Three (9.09%) out of 33 samples were positive in Timiș County, and one (5.26%) out of 19 samples was positive in Arad County. Subsequent PCR testing revealed that three samples were positive for Calicophoron daubneyi and one for Paramphistomum leydeni. The presence of C. daubneyi and P. leydeni in roe deer has not been previously reported in Romania. Two Paramphistomum species-C. daubneyi and P. leydeni-were revealed as the main species of rumen flukes in roe deer from forests in Romania.

[The importance of MR imaging in the diagnosis and management of acute stroke]. / Az MR-képalkotás jelentosége az akut stroke diagnosztikájában és ellátásában ­ Az MRI FIRST! Projekt.

Background and purpose:

In our collaborative project, called MRI First!, every patient arriving with neurological symptoms of acute stroke and without contraindications was examined by MRI. Our aim was to detect the symptomatic lesions, to obtain appropriate information about the brain parenchyma and to analyse parenchymal perfusion and brain vasculature.

The examinations were conduc­ted on a Philips Ingenia 1.5 Tesla scanner with the following protocol: DWI-ADC, FLAIR, T2 FFE/SWI, PWI, and contrast-enhanced MRA. 415 patients were examined between January 2020 and May 2021. 179 patients arrived within-, and 136 patients after 4.5 hours symptoms onset time, while 100 patients had “wake-up” stroke.

Within the 4.5 hours group, 81 cases had acute ischemic lesion, 48 of them received reperfusion therapy. Acute ische­mic lesion was found in 64 patients in the wake-up stroke group and in 64 in the 4.5-24 hours group. In these groups 10 and 12 patients obtained reperfusion therapy, respectively. Further 117 cases were considered as stroke mimics, in which cases unnecessary intravenous thrombolysis was avoidable.

MRI is accepted as a sensitive diagnostic modality providing detailed information regarding the brain parenchyma, its perfusion and vasculature. Nonetheless, its worldwide utilization in acute stroke is low and further information should be collected on which patient groups would gain the most benefit from acute MR imaging. Our continuous work is aimed at that goal.

Antimicrobial resistance gene lack in tick-borne pathogenic bacteria.

Tick-borne infections, including those of bacterial origin, are significant public health issues. Antimicrobial resistance (AMR), which is one of the most pressing health challenges of our time, is driven by specific genetic determinants, primarily by the antimicrobial resistance genes (ARGs) of bacteria. In our work, we investigated the occurrence of ARGs in the genomes of tick-borne bacterial species that can cause human infections. For this purpose, we processed short/long reads of 1550 bacterial isolates of the genera Anaplasma (n = 20), Bartonella (n = 131), Borrelia (n = 311), Coxiella (n = 73), Ehrlichia (n = 13), Francisella (n = 959) and Rickettsia (n = 43) generated by second/third generation sequencing that have been freely accessible at the NCBI SRA repository. From Francisella tularensis, 98.9% of the samples contained the FTU-1 beta-lactamase gene. However, it is part of the F. tularensis representative genome as well. Furthermore, 16.3% of them contained additional ARGs. Only 2.2% of isolates from other genera (Bartonella: 2, Coxiella: 8, Ehrlichia: 1, Rickettsia: 2) contained any ARG. We found that the odds of ARG occurrence in Coxiella samples were significantly higher in isolates related to farm animals than from other sources. Our results describe a surprising lack of ARGs in these bacteria and suggest that Coxiella species in farm animal settings could play a role in the spread of AMR.

ß-Tricalcium Phosphate-Modified Aerogel Containing PVA/Chitosan Hybrid Nanospun Scaffolds for Bone Regeneration.

Electrospinning has recently been recognized as a potential method for use in biomedical applications such as nanofiber-based drug delivery or tissue engineering scaffolds. The present study aimed to demonstrate the electrospinning preparation and suitability of ß-tricalcium phosphate-modified aerogel containing polyvinyl alcohol/chitosan fibrous meshes (BTCP-AE-FMs) for bone regeneration under in vitro and in vivo conditions. The mesh physicochemical properties included a 147 ± 50 nm fibrous structure, in aqueous media the contact angles were 64.1 ± 1.7°, and it released Ca, P, and Si. The viability of dental pulp stem cells on the BTCP-AE-FM was proven by an alamarBlue assay and with a scanning electron microscope. Critical-size calvarial defects in rats were performed as in vivo experiments to investigate the influence of meshes on bone regeneration. PET imaging using 18F-sodium fluoride standardized uptake values (SUVs) detected 7.40 ± 1.03 using polyvinyl alcohol/chitosan fibrous meshes (FMs) while 10.72 ± 1.11 with BTCP-AE-FMs after 6 months. New bone formations were confirmed by histological analysis. Despite a slight change in the morphology of the mesh because of cross-linking, the BTCP-AE-FM basically retained its fibrous, porous structure and hydrophilic and biocompatible character. Our experiments proved that hybrid nanospun scaffold composite mesh could be a new experimental bone substitute bioactive material in future medical practice.

A survey on antimicrobial resistance genes of frequently used probiotic bacteria, 1901 to 2022.

BackgroundAntimicrobial resistance (AMR) is caused by AMR determinants, mainly genes (ARGs) in the bacterial genome. Bacteriophages, integrative mobile genetic elements (iMGEs) or plasmids can allow ARGs to be exchanged among bacteria by horizontal gene transfer (HGT). Bacteria, including bacteria with ARGs, can be found in food. Thus, it is conceivable that in the gastrointestinal tract, bacteria from the gut flora could take up ARGs from food.AimThe study objective was to gain insight into the ARG set carried by commonly used probiotic bacteria that may enter the human body with non-fermented foods, fermented foods, or probiotic dietary supplements (FFPs) and to assess ARG mobility.MethodsNext generation sequencing whole genome data from 579 isolates of 12 commonly employed probiotic bacterial species were collected from a public repository. Using bioinformatical tools, ARGs were analysed and linkage with mobile genetic elements assessed.ResultsResistance genes were found in eight bacterial species. The ratios of ARG positive/negative samples per species were: Bifidobacterium animalis (65/0), Lactiplantibacillus plantarum (18/194), Lactobacillus delbrueckii (1/40), Lactobacillus helveticus (2/64), Lactococcus lactis (74/5), Leucoconstoc mesenteroides (4/8), Levilactobacillus brevis (1/46), Streptococcus thermophilus (4/19). In 66% (112/169) of the ARG-positive samples, at least one ARG could be linked to plasmids or iMGEs. No bacteriophage-linked ARGs were found.ConclusionThe finding of potentially mobile ARGs in probiotic strains for human consumption raises awareness of a possibility of ARG HGT in the gastrointestinal tract. In addition to existing recommendations, screening FFP bacterial strains for ARG content and mobility characteristics might be considered.

First detection of Ixodiphagus hookeri (Hymenoptera: Encyrtidae) in Ixodes ricinus ticks (Acari: Ixodidae) from multiple locations in Hungary.

The parasitoid wasp, Ixodiphagus hookeri (Hymenoptera: Encyrtidae), is the natural enemy of a wide range of hard and soft tick species. While these encyrtid wasps are supposed to be distributed worldwide, only a few studies report on their actual distribution around the globe. Within a shotgun sequencing-based metagenome analysis, the occurrence of I. hookeri was screened at multiple Ixodes ricinus (Acari: Ixodidae) tick sampling points in Hungary to contribute to the assessment of the distribution patterns of the parasitoid wasps in Central Europe. To our knowledge, the first report of the species in Hungary and the description of the southernmost I. hookeri associated geoposition in Central Europe took place within our study. I. hookeri infested I. ricinus nymphs were detected at five sampling points in Hungary. The results show that the exact distribution range of I. hookeri is still barely studied. At the same time, unprecedented public health issues being brought about by climate change might require steps toward the exploitation of the tick biocontrol potential and as an ecological bioindicator role of the parasitoid wasp in the future.

Canine Saliva as a Possible Source of Antimicrobial Resistance Genes.

While the One Health issues of intensive animal farming are commonly discussed, keeping companion animals is less associated with the interspecies headway of antimicrobial resistance. With the constant advance in veterinary standards, antibiotics are regularly applied in companion animal medicine. Due to the close coexistence of dogs and humans, dog bites and other casual encounters with dog saliva (e.g., licking the owner) are common. According to our metagenome study, based on 26 new generation sequencing canine saliva datasets from 2020 and 2021 reposited in NCBI SRA by The 10,000 Dog Genome Consortium and the Broad Institute within Darwin's Ark project, canine saliva is rich in bacteria with predictably transferable antimicrobial resistance genes (ARGs). In the genome of potentially pathogenic Bacteroides, Capnocytophaga, Corynebacterium, Fusobacterium, Pasteurella, Porphyromonas, Staphylococcus and Streptococcus species, which are some of the most relevant bacteria in dog bite infections, ARGs against aminoglycosides, carbapenems, cephalosporins, glycylcyclines, lincosamides, macrolides, oxazolidinone, penams, phenicols, pleuromutilins, streptogramins, sulfonamides and tetracyclines could be identified. Several ARGs, including ones against amoxicillin-clavulanate, the most commonly applied antimicrobial agent for dog bites, were predicted to be potentially transferable based on their association with mobile genetic elements (e.g., plasmids, prophages and integrated mobile genetic elements). According to our findings, canine saliva may be a source of transfer for ARG-rich bacteria that can either colonize the human body or transport ARGs to the host bacteriota, and thus can be considered as a risk in the spread of antimicrobial resistance.

Comparison of techno-functional and sensory properties of sponge cakes made with egg powder and different quality of powdered blood products for substituting egg allergen and developing functional food.

Animal blood is a valuable resource, which is usually not utilized in a value-added way by the industry like other animal by-products, even though it has plenty of benefits in terms of sustainability and human health, particularly against iron deficiency anemia. Animal blood is perfectly suitable for providing special functions, which are necessary for functional foods, and improving techno-functional properties based on the previous reports published in the literature. In this paper, egg powder was substituted by powdered animal blood products (whole blood powder, blood plasma powder, and hemoglobin powder) in sponge cake. Techno-functional and sensory properties (texture by texture profile analysis and three-point breaking test, water activity, dry matter content, and color) were instrumentally measured and then a sensory evaluation was carried out by unskilled panelists. Quality characteristics (texture, color, and dry matter content) were daily measured on the day of baking and then every 24 h for 3 additional days because freshly baked cakes are usually consumed within 3 days. Based on the results, powdered blood products are suitable for substituting the egg powder in sponge cakes and developing functional foods. Blood powders can increase the hardness, chewiness, and breaking force of cakes, giving them the ability to be stuffed with more fillings and molded into special shapes without compromising on the sensory characteristics. They can also increase the intensity of the cocoa flavor, which results in a richer, darker color without deceiving the consumers.

Exploring Relationships among Grapevine Chemical and Physiological Parameters and Leaf and Berry Mycobiome Composition.

Improving our knowledge on biotic and abiotic factors that influence the composition of the grapevine mycobiome is of great agricultural significance, due to potential effects on plant health, productivity, and wine characteristics. Here, we assessed the influence of scion cultivar on the diversity and composition of fungal communities in the berries and leaves of three different cultivars. We generated DNA metabarcoding data, and statistically compared the richness, relative abundance, and composition of several functional groups of fungi among cultivars, which are partly explained by measured differences in chemical composition of leaves and berries and physiological traits of leaves. Fungal communities in leaves and berries show contrasting patterns among cultivars. The richness and relative abundance of fungal functional groups statistically differ among berry and leaf samples, but less so among cultivars. Community composition of the dominant functional groups of fungi, i.e., plant pathogens in leaves and saprotrophs in berries, differs significantly among cultivars. We also detect cultivar-level differences in the macro- and microelement content of the leaves, and in acidity and sugar concentration of berries. Our findings suggest that there appears to be a relatively diverse set of fungi that make up the grapevine mycobiome at the sampled terroir that spans several cultivars, and that both berry and leaf mycobiomes are likely influenced by the chemical characteristics of berries and leaves, e.g., pH and the availability of nutrients and simple carbohydrates. Finally, the correlation between fungal community composition and physiological variables in leaves is noteworthy, and merits further research to explore causality. Our findings offer novel insights into the microbial dynamics of grapevine considering plant chemistry and physiology, with implications for viticulture.

Synthesis of New Type Polymers by Quasi-Living Atom Transfer Radical Polymerization.

Thanks to the polymer revolution of the 20th century, plastics are now part of our everyday lives. We use plastics as naturally as if they had always been an integral part of our lives. However, in the recent past, we were still predominantly using wood, metal, and glass objects, which were replaced by plastic products at an explosive rate. In many cases, this replacement has resulted in products with better physical, chemical, or biological properties. The changeover was too rapid, and the consequences were not recognized in time. This is evidenced by the huge scale of plastic pollution worldwide today. It is therefore in the interests of the future of both humans and animals that we must pay particular attention to the direct and indirect environmental impact of plastics introduced in animal husbandry. Starting from the tetrafunctional initiator produced as the first step of my work, poly(n-butyll acrylate) star polymers of different molecular weights were synthesized by atom transfer radical polymerization, using the so-called "core first" method. The bromine chain end of the produced star polymers was replaced by an azide group using a substitution reaction. Propalgyl telechelic PEGs were synthesized as a result of lattice end modification of poly(ethylene glycol) with different molecular weights. The azidated star polymers were connected with propalgyl telechelic PEGs using Huisgen's "click" chemical process, and as a result of the "click" connection, amphiphilic polymer networks with several different structures were obtained.

Antimicrobial resistance determinants in silage.

Animal products may play a role in developing and spreading antimicrobial resistance in several ways. On the one hand, residues of antibiotics not adequately used in animal farming can enter the human body via food. However, resistant bacteria may also be present in animal products, which can transfer the antimicrobial resistance genes (ARG) to the bacteria in the consumer's body by horizontal gene transfer. As previous studies have shown that fermented foods have a meaningful ARG content, it is indicated that such genes may also be present in silage used as mass feed in the cattle sector. In our study, we aspired to answer what ARGs occur in silage and what mobility characteristics they have? For this purpose, we have analyzed bioinformatically 52 freely available deep sequenced silage samples from shotgun metagenome next-generation sequencing. A total of 16 perfect matched ARGs occurred 54 times in the samples. More than half of these ARGs are mobile because they can be linked to integrative mobile genetic elements, prophages or plasmids. Our results point to a neglected but substantial ARG source in the food chain.

Investigation of the effects of bovine collagen peptides and mixed berries on rheological properties and biological activity of egg white-based beverage via central composite design.

Modern consumer expectations have become highly diversified: they want more opportunities to meet diverse family needs (diversity of family members in age, gender, physical activity, etc. ,) and individual health goals with a huge variety of sensorial preferences. Our research is aimed to develop a protein-dense, highly bioactive, lactose- and whey protein-free beverage applying a central composite rotational design (CCRD) with 2 factors. For this purpose, an egg white-based beverage was flavored with mixed berries (factor A) and enriched with bovine collagen peptides (factor B). After suitable sample preparation, the rheological properties were investigated by an Anton Paar MCR 92 rheometer (with CC 27 system, and flow behavior was analyzed with a Herschel-Bulkley (H-B) model). The antioxidant capacity of samples was investigated by Ferric Reducing Antioxidant Power (FRAP) method, the total anthocyanin content was estimated based on a spectrophotometric method, and the total phenolic content was determined by the Folin Ciocalteu method. Our results are figured on response surfaces demonstrating that both factors and their interactions show a positive correlation with the examined parameters. Based on the CCRD, all investigated parameters are significantly influenced by at least one aspect and can be adequately estimated for further product development.

Mobile Antimicrobial Resistance Genes in Probiotics.

Even though people worldwide tend to consume probiotic products for their beneficial health effects on a daily basis, recently, concerns were outlined regarding the uptake and potential intestinal colonisation of the bacteria that they carry. These bacteria are capable of executing horizontal gene transfer (HGT) which facilitates the movement of various genes, including antimicrobial resistance genes (ARGs), among the donor and recipient bacterial populations. Within our study, 47 shotgun sequencing datasets deriving from various probiotic samples (isolated strains and metagenomes) were bioinformatically analysed. We detected more than 70 ARGs, out of which rpoB mutants conferring resistance to rifampicin, tet(W/N/W) and potentially extended-spectrum beta-lactamase (ESBL) coding TEM-116 were the most common. Numerous ARGs were associated with integrated mobile genetic elements, plasmids or phages promoting the HGT. Our findings raise clinical and public health concerns as the consumption of probiotic products may lead to the transfer of ARGs to human gut bacteria.