Ethyl glucuronide and ethyl sulfate in the zebrafish after ethanol exposure.

Ethanol exposure during pregnancy is an important problem and is the cause of fetal alcohol syndrome (FAS) and fetal alcohol spectrum disorder (FASD). The etiology of FAS and FASD can be elucidated using animal models. Recently, a novel model, the zebrafish (Danio rerio), has garnered the interest of researchers. This study confirmed the negative influence of ethyl alcohol (0.5 %, 1.5 %, and 2.5 % v/v) on the development of zebrafish embryos. The observed malformations included pericardial and yolk sac edema, increased body curvature, tail edema, and a decreased embryo hatching rate. The differences in body length, body width, and heart rate were statistically significant. Due to the similarities in the quantity and function of ethanol biotransformation enzymes between zebrafish and mammals, this study investigated the nonoxidative metabolites of ethanol - ethyl glucuronide (EtG) and ethyl sulfate (EtS) - in zebrafish following ethanol exposure. This research confirmed that EtG and EtS concentrations can be measured in zebrafish embryos, and the levels of these metabolites appear to be associated with the ethyl alcohol concentration in the medium.

Variation of Sequential Ligandrol (LGD-4033) Metabolite Levels in Routine Anti-Doping Urine Samples Detected with or without Other Xenobiotics.

Ligandrol, also known as LGD-4033, belongs to the group of selective androgen receptor modulators (SARMs). Ligandrol was first included in the WADA Prohibited List in 2018. This work presents a method that allows for the detection and identification of ligandrol and its metabolite in athletes' urine and in dietary supplements by means of ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Samples were prepared according to an approach involving acid hydrolysis and double liquid-liquid extraction (LLE). Furthermore, due to the lack of reference material for ligandrol metabolites, the urine collected from the control excretion study was analyzed. The presented method is appropriate to monitor ligandrol and its metabolites. The samples collected for doping control purpose contained multiple metabolites, which may potentially rule out the hypothesis of ingesting a single 1 µg or 10 µg dose only. Another aspect to take into account is that ligandrol can be applied together with SARMs, steroids, and GHSs. This will also affect the substances' metabolism and elimination. It is also worth noting that dietary supplements may contain ligandrol as an official ingredient or as a contaminant. The described method may be usefully applied by other anti-doping or toxicological laboratories.

Spatial consistency of cell growth direction during organ morphogenesis requires CELLULOSE SYNTHASE INTERACTIVE1.

Extracellular matrices contain fibril-like polymers often organized in parallel arrays. Although their role in morphogenesis has been long recognized, it remains unclear how the subcellular control of fibril synthesis translates into organ shape. We address this question using the Arabidopsis sepal as a model organ. In plants, cell growth is restrained by the cell wall (extracellular matrix). Cellulose microfibrils are the main load-bearing wall component, thought to channel growth perpendicularly to their main orientation. Given the key function of CELLULOSE SYNTHASE INTERACTIVE1 (CSI1) in guidance of cellulose synthesis, we investigate the role of CSI1 in sepal morphogenesis. We observe that sepals from csi1 mutants are shorter, although their newest cellulose microfibrils are more aligned compared to wild-type. Surprisingly, cell growth anisotropy is similar in csi1 and wild-type plants. We resolve this apparent paradox by showing that CSI1 is required for spatial consistency of growth direction across the sepal.

Plant biology: How the humble plant droops its leaves.

The humble plant (Mimosa pudica) droops its leaves in response to touch. A new study explains how changes of turgor pressure exerted by protoplasts on surrounding cell walls translate into directional cell deformation that drives leaf movement.

Digestibility of Protein and Iron Availability from Enriched Legume Sprouts.

Plant ferritin is suggested as a good source of iron for human. Usually present in trace amounts, it was induced in legumes seeds by their sprouting in FeSO4 solution. Fortified sprouts were digested in the in vitro model of the human gastrointestinal tract. ~49% of lupine and ~ 45% of soy proteins were extracted into gastric fluid and next ~ 12% and only ~ 1% into intestine fluid from lupine and soybean, respectively. Gastric digestion released mainly ferrous iron (~ 85% from lupine and ~ 95% in soybean sprouts). Complexed iron constituted ~ 43% of total iron in intestine after lupine digestion and ~ 55% after soybean digestion. Intestine digestion doubled the total iron released from lupine sprouts (from ~ 21% up to 38%), while in soybean it increased from ~ 16% up to ~ 23%. Ferritin presence was confirmed by the specific antibodies in digestive fluids, but it is only partially extracted from sprouts during in vitro digestion.

Ecdysterone: Possible sources of origin in urine.

Ecdysterone (crustecdysone; beta-ecdysone; 20-hydroxyecdysone) is a naturally occurring steroid hormone belonging to the ecdysteroid class. The presented study investigated the possible concentration range of ecdysterone in urine after consumption of various preparations of spinach, drinking tea (made from Rhapoiticum Carthamoides) and topical use of a cream containing Cyanotis arachnoides. It is very important to establish reference ranges reflecting concentrations compatible with dietary habits and common uses of care products. The data obtained in the research may be used in the interpretation of results of routine analyses. In addition, elimination time and observed concentrations provided by the studies conducted by the Polish Anti-Doping Laboratory can be used by WADA. In the case of spinach, peak elimination occurred within the first few hours, followed by a rapid decline. As for the other plants, instead of clear peak concentrations, gradual elimination was observed. Individual differences were observed between volunteers depending on route of administration. Differences in ecdysterone elimination following ingestion of spinach-based and other plant products were observed too. The highest observed ecdysterone concentration was related to the paste consumption, and it was 691 ng/ml. Finally, our findings were compared with the data collected for the samples routinely tested as part of the monitoring program. During 2.5 years, the presence of ecdysterone was confirmed in as many as 507 samples out of 11 191 total samples tested. The concentration range was very wide, from 1 ng/ml (which is the LOD for this method in the Polish Anti-Doping Laboratory) to over 2000 ng/ml.

Plant Biomechanics-A Natural Transition from Molecular to Organ Scale.

Plants are multicellular organisms of a unique structure because their tissues consist of two interwoven networks: a network of interconnected protoplasts that is embedded in a network of tightly joined cell walls [...].

Elite athletes with COVID-19 - Predictors of the course of disease.

OBJECTIVES: The aim of the study was to identify predictors determining the course of COVID-19 and antibody response in elite athletes. DESIGN: Observational study. METHODS: Routine medical screening with physical examination, resting ECG, and laboratory tests including antibody response was performed 12-68 days after the diagnosis of COVID-19 in 111 athletes of different sports. RESULTS: Clinical symptoms were observed in 84% of subjects. The severity of COVID-19 was mild in 82% of athletes and moderate in 2% of cases. Athletes aged above 26 and male were more likely to develop symptomatic COVID-19. Asymptomatic subjects were younger and predominantly female. In 18% of subjects, symptoms were still present 20 (12-68) days (median and range) since positive diagnosis. Antibody response was observed in 88% of athletes, and its magnitude correlated with time since diagnosis of COVID-19 (RT-PCR), fatigue, fever, and conjunctivitis. There were no differences in antibody response between groups distinguished by sports discipline (p = 0.50), and sex (p = 0.59), and antibody response did not correlate with BMI (p = 0.12), age (p = 0.13), the number of symptoms (p = 0.43) or their duration (p = 0.19). CONCLUSIONS: The severity of COVID-19 in elite athletes is predominantly mild and without complications. Athletes can return to sport after two symptom-free weeks and additional heart screening is usually not required. Determination of antibodies has been shown to be a useful indicator of a previous COVID-19 disease, and some symptoms can be used as predictors of antibody response.

A Review of Honeybee Venom Allergens and Allergenicity.

Honeybee venom is a source of proteins with allergenic properties which can result in in various symptoms, ranging from local reactions through to systematic life-threatening anaphylaxis, or even death. According to the World Allergy Organization (WAO), honeybee venom allergy is one of the most common causes of anaphylaxis. Among the proteins present in honeybee venom, 12 protein fractions were registered by the World Health Organization's Allergen Nomenclature Sub-Committee (WHO/IUIS) as allergenic. Most of them are highly immunogenic glycoproteins that cross-react with IgE and, as a consequence, may give false positive results in allergy diagnosis. Allergenic fractions are different in terms of molecular weight and biological activity. Eight of these allergenic fractions have also been identified in honey. This explains frequent adverse reactions after consuming honey in people allergic to venom and sheds new light on the causes of allergic symptoms in some individuals after honey consumption. At the same time, it also indicates the possibility of using honey as a natural source of allergen in specific immunotherapy.

Apple allergy: Causes and factors influencing fruits allergenic properties-Review.

BACKGROUND: Apple tree fruits (Malus × domestica Borkh.) are a rich source of nutrients and nutraceuticals and are recommended as a part of the healthy, staple diet. However, apples could be also the cause of allergies including severe reactions. Allergies to fruits like apples are predominantly associated with pollinosis. In North and Central Europe, sensitisation to apples is caused mainly by cross-reactive birch pollen aeroallergen, whereas in the Mediterranean area of Europe, apple allergy is mostly associated with allergies to peach. The allergenicity of apples differ across cultivars but only a few varieties were studied. Some factors changing apples allergenicity were identified, including unmodifiable and potentially modifiable factors for example cultivation method, ripening stage and storage conditions. AIM: This review presents current knowledge about the molecular basis of apple allergenicity and factors influencing its level. CONCLUSIONS: Selecting cultivars with low potential of allergenicity, removing apple peel and heat treatment could reduce the risk of severe allergy reaction incidence and presumably can be used in birch pollen immunotherapy.

Molecular and Immunological Identification of Low Allergenic Fruits among Old and New Apple Varieties.

About 50-70% of patients allergic to birch pollen suffer from sensitization after apple ingestion. Apple allergenicity was established in only few varieties. Studies were performed on apple fruits of 21 traditional and nine modern varieties organically, intensively, or integratively produced. The aim of the study was to assess whether the factors like cultivation method, maturity stage, genotype, or type of tissue place an impact on the allergenic potential of apples. To answer these questions, we used semiquantitative real-time PCR, ELISA, and immunoblotting. Apple allergen genes present divergent expression across apple cultivars. Expression of the Mal d 1.06A correlates with the Mal d 1 level and is affected by the cultivation method and maturity of the fruit. The content of the main allergen Mal d 1 varied widely across cultivars. Interestingly, in our study, the Gala variety presented a low Mal d 1 concentration regardless of the cultivation method. Based on the Mal d 1.06A expression, the Mal d 1 protein content, and the immunoreactivity assay, the Kandil Sinap, Kosztela, Rumianka from Alma-Ata, Kantówka Gdanska, Reinette Coulon, and Gala cultivars emerged as potentially hypoallergenic apple cultivars. Our study allowed distinguishing between potentially low, medium, and highly allergenic varieties.

Structure, Assembly and Function of Cuticle from Mechanical Perspective with Special Focus on Perianth.

This review is devoted to the structure, assembly and function of cuticle. The topics are discussed from the mechanical perspective and whenever the data are available a special attention is paid to the cuticle of perianth organs, i.e., sepals, petals or tepals. The cuticle covering these organs is special in both its structure and function and some of these peculiarities are related to the cuticle mechanics. In particular, strengthening of the perianth surface is often provided by a folded cuticle that functionally resembles profiled plates, while on the surface of the petal epidermis of some plants, the cuticle is the only integral continuous layer. The perianth cuticle is distinguished also by those aspects of its mechanics and development that need further studies. In particular, more investigations are needed to explain the formation and maintenance of cuticle folding, which is typical for the perianth epidermis, and also to elucidate the mechanical properties and behavior of the perianth cuticle in situ. Gaps in our knowledge are partly due to technical problems caused by very small thicknesses of the perianth cuticle but modern tools may help to overcome these obstacles.

Xylan Decomposition in Plant Cell Walls as an Inducer of Surfactin Synthesis by Bacillus subtilis.

Hemicellulose is the second most abundant plant heterogenous biopolymer. Among products obtained from a wide range of agro-residues, biosurfactants, e.g., surfactin (SU), are gaining increasing interest. Our previous studies have shown that a Bacillus subtilis strain can successfully produce a significant amount of SU using a rapeseed cake. This work aimed to investigate plant hemicellulose components as substrates promoting SU's efficient production by B. subtilis 87Y. Analyses of SU production, enzymatic activity and cell wall composition of hulled oat caryopses suggest that the main ingredients of plant hemicellulose, in particular xylan and its derivatives, may be responsible for an increased biosurfactant yield.

What is quantitative plant biology?

Quantitative plant biology is an interdisciplinary field that builds on a long history of biomathematics and biophysics. Today, thanks to high spatiotemporal resolution tools and computational modelling, it sets a new standard in plant science. Acquired data, whether molecular, geometric or mechanical, are quantified, statistically assessed and integrated at multiple scales and across fields. They feed testable predictions that, in turn, guide further experimental tests. Quantitative features such as variability, noise, robustness, delays or feedback loops are included to account for the inner dynamics of plants and their interactions with the environment. Here, we present the main features of this ongoing revolution, through new questions around signalling networks, tissue topology, shape plasticity, biomechanics, bioenergetics, ecology and engineering. In the end, quantitative plant biology allows us to question and better understand our interactions with plants. In turn, this field opens the door to transdisciplinary projects with the society, notably through citizen science.

The analytical approach for detection of carbamylated erythropoietin for doping control purposes.

Erythropoietin (EPO) has protective effects in several tissues and could be used for therapeutic purposes, but the doses of EPO that can be beneficial in case of hypoxic-ischemic conditions due to overinduced erythropoiesis could be detrimental in treated patients. Carbamylation of erythropoietin maintains the tissue-protective effects of EPO but without erythropoietic effects. Carbamylated EPO (CEPO) is listed in WADA Prohibited List in class S2 as "Innate repair receptor agonists." The CEPO was synthesized using the method described previously. Digestion with endoproteinase Lys-C was used to distinguish rhEPO from CEPO. The digested samples containing recombinant EPO, urinary EPO (uEPO), or CEPO were analyzed by the SAR-PAGE method (sarcosyl polyacrylamide gel electrophoresis-PAGE). Endoproteinase Lys-C breaks the peptide chains of lysine. Lysine residues, converted to homocitrulline by carbamylation, cannot be cleaved by endoproteinase Lys-C. Therefore, the CEPO protein chain remained unchanged in contrast to rhEPO and uEPO, which allows for easily differentiation of them.

Allergenic properties of Polish nectar honeys.

BACKGROUND: Small amounts of protein can be found in honey, including well known allergen sources, such as plant pollen and honeybee secretions. Despite this, there are few case reports describing allergic reactions following the consumption of honey. The aim of this study was to examine the allergenic properties of nectar honey collected throughout the entire beekeeping season from different provinces in Poland. METHODS: The immunoreactive properties of 20 Polish nectar honeys were analysed using the sera of IgE pollen allergenic patients (n = 5). The botanical origins and pollen of the anemophilous plants in the studied honeys were identified through palynological analysis. RESULTS: The significant differences in the protein content between the five varieties of honey and the differences in protein pattern and pollen profiles were observed. All of the honey samples contained immunoreactive fractions reacting with IgE present in the sera of patients allergenic to different pollens. CONCLUSIONS: Although honey allergies are reported relatively rarely, all the tested samples of Polish nectar honeys contained many protein fractions which reacted with the IgE antibodies of allergenic patients. In all samples, the immunoreactive protein band with a molecular weight around 60 kDa, probably secreted by bees, was present. The results do not allow the immunoreactive fractions characteristic for particular honey varieties to be identified.

Soy Preparations Are Potentially Dangerous Factors in the Course of a Food Allergy.

The special properties of soy preparations make them common additives for food production and can be dangerous for sensitive individuals. Our aim was to check consumers' awareness of potential risks of soy preparations added to numerous food products, depending on respondents' education, and to evaluate immunoreactive properties of chosen soy preparations. A personal questionnaire was used. Respondents (n = 251) were aged 23-28 years old, lived in Poland, and were graduates or students in their last year of food technology, medicine, and university of technology. The slot blot and Western blotting methods were used to mark immunoreactivity of soy preparations. It was shown that most respondents often or usually read labels of food products they buy. The surveyed indicated protein is the allergenic component in soy. Almost half of them were of the opinion that hydrolysis removes the allergenic properties of soy. Most of the medical students surveyed thought that people allergic to soy may consume products that contain soy preparations. The analytical results indicated that soy preparation contained protein fractions that were immunoreactive with sera of allergenic patients. It was proven that preparations, even hydrolysates, contain immunoreactive proteins that may be the source of hidden allergens, even though they are not recognized as dangerous by well-educated respondents.

Sequential Replicas: Method for In Vivo Imaging of Plant Organ Surfaces that Undergo Deformation.

Complex geometry of plant organs and various types of organ surface deformation, including growth or hygroscopic movements, can be analyzed using sequential replica method. It enables obtaining a time-lapse series of high resolution images visualizing details of the examined surface and provides data sufficient for detailed computation of parameters characterizing surface deformation and geometry. Series of molds, made in dental polymer, representing the examined surface are used to obtain casts in epoxy resin or nail polish replicas, which are ready for microscopic examination, while the structure itself remains intact. Images obtained from the epoxy casts in scanning electron microscopy can be further used for 3D reconstruction and computation of local geometry. The sequential replica method is a universal method and can be applied to image complex shapes of a range of structures, like meristems, flowers, leaves, scarious bracts, or trichomes. Different plant species growing in various conditions can be studied.