Quality Assessment of Reconstructed Cow, Camel and Mare Milk Powders by Near-Infrared Spectroscopy and Chemometrics.

Milk powders are becoming a major attraction for many industrial applications due to their nutritional and functional properties. Different types of powdered milk, each with their own distinct chemical compositions, can have different functionalities. Consequently, the development of rapid monitoring methods is becoming an urgent task to explore and expand their applicability. Lately, there is growing emphasis on the potential of near-infrared spectroscopy (NIRS) as a rapid technique for the quality assessment of dairy products. In the present work, we explored the potential of NIRS coupled with chemometrics for the prediction of the main functional and chemical properties of three types of milk powders, as well as their important processing parameters. Mare, camel and cow milk powders were prepared at different concentrations (5%, 10% and 12%) and temperatures (25 °C, 40 °C and 65 °C), and then their main physicochemical attributes and NIRS spectra were analyzed. Overall, high accuracy in both recognition and prediction based on type, concentration and temperature was achieved by NIRS-based models, and the quantification of quality attributes (pH, viscosity, dry matter content, fat content, conductivity and individual amino acid content) also resulted in high accuracy in the models. R2CV and R2pr values ranging from 0.8 to 0.99 and 0.7 to 0.98, respectively, were obtained by using PLSR models. However, SVR models achieved higher R2CV and R2pr values, ranging from 0.91 to 0.99 and 0.80 to 0.99, respectively.

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.

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.

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.

[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.

Follistatin treatment suppresses SERCA1b levels independently of other players of calcium homeostasis in C2C12 myotubes.

Follistatin (FS) is a high affinity activin-binding protein, neutralizing the effects of the Transforming Growth Factor-beta (TGF-ß) superfamily members, as myostatin (MSTN). Since MSTN emerged as a negative regulator, FS has been considered as a stimulator of skeletal muscle growth and differentiation. Here, we studied the effect of FS administration on the Ca2+-homeostasis of differentiating C2C12 skeletal muscle cells. FS-treatment increased the fusion index, the size of terminally differentiated myotubes, and transiently elevated the expression of the calcium-dependent protein phosphatase, calcineurin, at the beginning of differentiation. Functional experiments did not detect any alterations in the Ca2+ transients following the stimulation by KCl or caffeine in myotubes. On the other hand, decreased Ca2+-uptake capability was determined by calculating the maximal pump rate (332 ± 17 vs. 279 ± 11 µM/s, in control and FS-treated myotubes, respectively; p < 0.05). In the same way, the expression and ATPase activity of the neonatal sarcoplasmic/endoplasmic reticulum Ca2+ATPase (SERCA1b) were decreased (0.59 ± 0.01 vs. 0.19 ± 0.01 mM ATP/min, in control and FS-treated myotubes, respectively; p < 0.05). However, the expression level of other proteins involved in Ca2+-homeostasis and differentiation (calsequestrin, STIM1, MyoD) were not affected. Our results suggest that the FS controlled myotube growth is paralleled with the tight regulation of cytosolic calcium concentration, and the decline of SERCA1b appears to be one of the key components in this process.

Wartime scars or reservoirs of biodiversity? The value of bomb crater ponds in aquatic conservation.

Considering the ongoing loss of aquatic habitats, anthropogenic ponds are gaining importance as substitute habitats. It is therefore important to assess their functioning in comparison to their natural precursors. Here we assess the biodiversity value of sodic bomb crater ponds by comparing their gamma diversity to that of natural reference habitats, astatic soda pans, and assess their importance on the landscape level by studying alpha and beta diversity. We studied aquatic organisms ranging from algae to vertebrates in a dense cluster of 54 sodic bomb crater ponds in Central Europe. Despite the overall small area of the pond cluster, gamma diversity was comparable to that found in surveys of natural habitats that encompassed much wider spatial and temporal scales. We also found a considerable number of species shared with reference habitats, indicating that these anthropogenic habitats function as important refuge sites for several species that are associated with the endangered soda pans. Moreover, we found a number of regionally or worldwide rare species. Among the components of beta diversity, species replacement dominated community assembly. Individual ponds contributed similarly to beta diversity in terms of replacement, being equally important for maintaining high gamma diversity and emphasising the role of the pond network rather than individual ponds. This pattern was seen in all studied groups. Bomb crater ponds therefore acted as important contributors to aquatic biodiversity. Considering the tremendous losses of ponds throughout Europe, anthropogenic ponds should be taken into consideration in nature conservation, especially when occurring in pond networks.

Cannabinoid signalling inhibits sarcoplasmic Ca2+ release and regulates excitation-contraction coupling in mammalian skeletal muscle.

KEY POINTS: Marijuana was found to cause muscle weakness, although the exact regulatory role of its receptors (CB1 cannabinoid receptor; CB1R) in the excitation-contraction coupling (ECC) of mammalian skeletal muscle remains unknown. We found that CB1R activation or its knockout did not affect muscle force directly, whereas its activation decreased the Ca2+ -sensitivity of the contractile apparatus and made the muscle fibres more prone to fatigue. We demonstrate that CB1Rs are not connected to the inositol 1,4,5-trisphosphate pathway either in myotubes or in adult muscle fibres. By contrast, CB1Rs constitutively inhibit sarcoplasmic Ca2+ release and sarcoplasmic reticulum Ca2+ ATPase during ECC in a Gi/o protein-mediated way in adult skeletal muscle fibres but not in myotubes. These results help with our understanding of the physiological effects and pathological consequences of CB1R activation in skeletal muscle and may be useful in the development of new cannabinoid drugs. ABSTRACT: Marijuana was found to cause muscle weakness, although it is unknown whether it affects the muscles directly or modulates only the motor control of the central nervous system. Although the presence of CB1 cannabinoid receptors (CB1R), which are responsible for the psychoactive effects of the drug in the brain, have recently been demonstrated in skeletal muscle, it is unclear how CB1R-mediated signalling affects the contraction and Ca²âº homeostasis of mammalian skeletal muscle. In the present study, we demonstrate that in vitro CB1R activation increased muscle fatigability and decreased the Ca2+ -sensitivity of the contractile apparatus, whereas it did not alter the amplitude of single twitch contractions. In myotubes, CB1R agonists neither evoked, nor influenced inositol 1,4,5-trisphosphate (IP3 )-mediated Ca2+ transients, nor did they alter excitation-contraction coupling. By contrast, in isolated muscle fibres of wild-type mice, although CB1R agonists did not evoke IP3 -mediated Ca2+ transients too, they significantly reduced the amplitude of the depolarization-evoked transients in a pertussis-toxin sensitive manner, indicating a Gi/o protein-dependent mechanism. Concurrently, on skeletal muscle fibres isolated from CB1R-knockout animals, depolarization-evoked Ca2+ transients, as well qas Ca2+ release flux via ryanodine receptors (RyRs), and the total amount of released Ca2+ was significantly greater than that from wild-type mice. Our results show that CB1R-mediated signalling exerts both a constitutive and an agonist-mediated inhibition on the Ca2+ transients via RyR, regulates the activity of the sarcoplasmic reticulum Ca2+ ATPase and enhances muscle fatigability, which might decrease exercise performance, thus playing a role in myopathies, and therefore should be considered during the development of new cannabinoid drugs.

Purinergic signalling is required for calcium oscillations in migratory chondrogenic progenitor cells.

Osteoarthritis (OA) is the most common form of chronic musculoskeletal disorders. A migratory stem cell population termed chondrogenic progenitor cells (CPC) with in vitro chondrogenic potential was previously isolated from OA cartilage. Since intracellular Ca(2+) signalling is an important regulator of chondrogenesis, we aimed to provide a detailed understanding of the Ca(2+) homeostasis of CPCs. In this work, CPCs immortalised by lentiviral administration of the human telomerase reverse transcriptase (hTERT) and grown in monolayer cultures were studied. Expressions of all three IP3Rs were confirmed, but no RyR subtypes were detected. Ca(2+) oscillations observed in CPCs were predominantly dependent on Ca(2+) release and store replenishment via store-operated Ca(2+) entry; CPCs express both STIM1 and Orai1 proteins. Expressions of adenosine receptor mRNAs were verified, and adenosine elicited Ca(2+) transients. Various P2 receptor subtypes were identified; P2Y1 can bind ADP; P2Y4 is targeted by UTP; and ATP may evoke Ca(2+) transients via detected P2X subtypes, as well as P2Y1 and P2Y2. Enzymatic breakdown of extracellular nucleotides by apyrase completely abrogated Ca(2+) oscillations, suggesting that an autocrine/paracrine purinergic mechanism may drive Ca(2+) oscillations in these cells. As CPCs possess a broad spectrum of functional molecular elements of Ca(2+) signalling, Ca(2+)-dependent regulatory mechanisms can be supposed to influence their differentiation potential.

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.

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.

Redox Homeostasis and Non-Invasive Assessment of Significant Liver Fibrosis by Shear Wave Elastography.

Hepatic fibrosis with various origins can be estimated non-invasively by using certain biomarkers and imaging-based measurements. The aim of our study was to examine redox homeostasis biomarkers and liver stiffness measurements for the assessment of significant liver fibrosis in different etiologies of chronic liver diseases. A cohort study consisting of 88 chronic liver disease patients of both sexes (age 49.1 ± 14.7 years) was performed. Cytokine profiles as well as redox homeostasis characteristics were determined. Liver fibrosis stages were assessed with shear wave elastography. The plasma levels of four cytokines showed no significant alteration between the four fibrotic stages; however, higher values were measured in the F2-4 stages. Free sulfhydryl group concentration, the marker of redox homeostasis, was lower in significant fibrosis (F0-F1: 0.36 ± 0.06 vs. F2-4: 0.29 ± 0.08 mmol/L, p < 0.05). Higher chemiluminescence values, as free radical-antioxidant parameters, were detected in advanced fibrosis stages in erythrocytes (F0-F1: 36.00 ± 37.13 vs. F2-4: 51.47 ± 44.34 RLU%). These data suggest that oxidative stress markers can predict significant fibrosis, with the aim of reducing the number of protocol liver biopsies in patients unlikely to have significant disease; however, their role in distinguishing between the certain fibrosis groups needs further studies.

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

BACKGROUND AND PURPOSE: The first-generation photon-counting detector 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 photon-counting detector CT compared with energy-integrating detector 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 energy-integrating detector CT were prospectively enrolled. CT angiograms were acquired on a clinical dual-source photon-counting detector CT in ultra-high-resolution mode and reconstructed with 4 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, contrast-to-noise ratio), ROIs were manually placed at standard anatomic intracranial and extracranial locations by 1 author. In addition, vessel border sharpness was evaluated quantitatively. For the qualitative analysis, 3 blinded neuroradiologists rated photon-counting detector CT and energy-integrating detector CT image quality for the evaluation of the intracranial vessels (ie, the aneurysms and 9 standard vascular branching locations) on a 5-point Likert-type scale. Additionally, readers independently selected their preferred kernel among the 4 kernels evaluated on photon-counting detector CT. RESULTS: In terms of quantitative image quality, Bv48, the sharpest kernel, yielded increased image noise and decreased SNR and contrast-to-noise ratio parameters compared with Bv36, the smoothest kernel. Compared with energy-integrating detector 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 energy-integrating detector CT and not significantly different from ratings of the B44 reconstruction kernel. When comparing side by side all 4 photon-counting detector reconstruction kernels, readers selected the B48 kernel as the best to visualize the aneurysms in 80% of cases. CONCLUSIONS: Ultra-high-resolution photon-counting detector CT provides improved image quality for neurovascular imaging. Although the less sharp kernels provided superior SNR and contrast-to-noise ratio, the sharpest kernels delivered the best subjective image quality on photon-counting detector CT for the evaluation of intracranial aneurysms.

Optimizing Quantum Iterative Reconstruction for Ultra-high-resolution Photon-counting Computed Tomography of the Lung.

PURPOSE: The aim of this study was to find the optimal strength level of QIR for ultra-high-resolution (UHR) PCCT of the lung. MATERIALS AND METHODS: This retrospective study included 24 patients who had unenhanced chest CT with the novel UHR scan protocol on the PCCT scanner between March 24, 2023 and May 18, 2023. Two sets of reconstructions were made using different slice thicknesses: standard resolution (SR, 1 mm) and ultra-high-resolution (UHR, 0.2 mm), reconstructed with all strength levels of QIR (0 to 4). Attenuation of the lung parenchyma, noise, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR) were assessed as objective criteria of image quality. Two fellowship-trained radiologists compared image quality and noise level, sharpness of the images, and the airway details using a 5-point Likert scale. Wilcoxon signed-rank test was used for statistical analysis of reader scores, and one-way repeated measures analysis of variance for comparing the objective image quality scores. RESULTS: Objective image quality linearly improved with higher strength levels of QIR, reducing image noise by 66% from QIR-0 to QIR-4 (P<0.001). Subjective image noise was best for QIR-4 (P<0.001). Readers rated QIR-1 and QIR-2 best for SR, and QIR-2 and QIR-3 best for UHR in terms of subjective image sharpness and airway detail, without significant differences between them (P=0.48 and 0.56, respectively). CONCLUSIONS: Higher levels of QIR provided excellent objective image quality, but readers' preference was for intermediate levels. Considering all metrics, we recommend QIR-3 for ultra-high-resolution PCCT of the lung.

Botrytis cinerea causes different plant responses in grape (Vitis vinifera) berries during noble and grey rot: diverse metabolism versus simple defence.

The complexity of the interaction between the necrotrophic pathogen Botrytis cinerea and grape berries (Vitis vinifera spp.) can result in the formation of either the preferred noble rot (NR) or the loss-making grey rot (GR), depending on the prevailing climatic conditions. In this study, we focus on the functional gene set of V. vinifera by performing multidimensional scaling followed by differential expression and enrichment analyses. The aim of this study is to identify the differences in gene expression between grape berries in the phases of grey rot, noble rot, and developing rot (DR, in its early stages) phases. The grapevine transcriptome at the NR phase was found to exhibit significant differences from that at the DR and GR stages, which displayed strong similarities. Similarly, several plant defence-related pathways, including plant-pathogen interactions as hypersensitive plant responses were found to be enriched. The results of the analyses identified a potential plant stress response pathway (SGT1 activated hypersensitive response) that was found to be upregulated in the GR berry but downregulated in the NR berry. The study revealed a decrease in defence-related in V. vinifera genes during the NR stages, with a high degree of variability in functions, particularly in enriched pathways. This indicates that the plant is not actively defending itself against Botrytis cinerea, which is otherwise present on its surface with high biomass. This discrepancy underscores the notion that during the NR phase, the grapevine and the pathogenic fungi interact in a state of equilibrium. Conversely the initial stages of botrytis infection manifest as a virulent fungus-plant interaction, irrespective of whether the outcome is grey or noble rot.

Evaluation of Gel Coating Performance in Extending the Shelf Life of Egg: The Role of Surface Area and Initial Weight.

This work investigated the impact of chicken egg size, including surface area and initial weight, on the effectiveness of cassava starch-based gel coating during storage at room temperature. The quality of a total of 540 fresh eggs in four different sizes (S, M, L and XL) was evaluated over a 4-week storage period at 25 ± 1 °C (60-65% RH). In this research, images from a scanning electron microscope revealed that the coatings maintained their integrity across all egg sizes, effectively covering pores and cracks throughout storage. The application of gel coating reduced weight loss and preserved the Haugh unit and yolk index, extending freshness by 1-2 weeks compared with uncoated eggs at 25 °C. The results indicated that the performance of the coating varied with egg size. Statistical analysis revealed that the surface area and initial weight of the egg significantly impacted the effectiveness of the coating in preserving quality (p < 0.001). Eggs with larger surface areas exhibited a reduced protective effect of the coating, resulting in higher weight loss and lower retention of Haugh unit and yolk index compared with the coated eggs with smaller surface areas. The coating application was more effective in preserving the Haugh unit of eggs with higher initial weights. Overall, the surface area and the initial weight of the egg should be considered as key factors to ensure optimal coating performance.

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.

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.

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.

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.