Black Queens of Fruits: Chemical Composition of Blackberry (Rubus subg. rubus Watson) and Black Currant (Ribes nigrum L.) Cultivars Selected in Serbia.

Black fruits, especially blackberries and black currants, are highly appreciated by consumers due to their nutraceutical properties, which have reported health benefits. This study aimed to assess the fruit quality of the blackberry (cv. 'Cacanska Bestrna') and black currant (cv. 'Cacanska Crna') created at the Fruit Research Institute, Cacak (Republic of Serbia) by evaluating basic quality parameters (fruit weight, soluble solids), and content of primary (sugars and organic acids) and secondary (phenolic compounds) metabolites. Additionally, the study examined the quality of frozen and long-term frozen storage (6, 9, and 12 months). The results showed that 'Cacanska Crna' contained a high level of total soluble solids (15.23°Bx), sugars (93.06 mg g-1), and a high index of sweetness (159.48) as well as a high content of acids (34.35 mg g-1) in the fruit. Both species were found to contain forty-seven phenolic compounds, including phenolic acids, flavanols, and anthocyanins, analyzed using high performance liquid chromatography-mass spectrometry (HPLC-MS). 'Cacanska Bestrna' had the highest amounts of phenolic acids (66.85 mg 100 g-1) and flavanols (53.99 mg 100 g-1), whereas 'Cacanska Crna' showed the highest levels of flavonols (8.57 mg 100 g-1) and anthocyanins (139.11 mg 100 g-1). Furthermore, the study revealed that anthocyanins were the most abundant phenolic group in both blackberries and black currants, and their levels remained constant during frozen storage. The sugar content in both species remained unchanged, while the acid concentration increased over time in blackberries but remained consistent in black currants. Overall, the findings highlight the superior phenolic content, particularly anthocyanins, of the cultivars with black fruits selected in Serbia. These cultivars have great commercial potential for the fresh market and processing. Moreover, the study suggests that frozen storage is an effective method for preserving their quality and beneficial properties.

Techno-Functional Performance of Emmer, Spelt and Khorasan in Spontaneously Fermented Sourdough Bread.

The aim of this study was to test the suitability of three different ancient wheat varieties (emmer, spelt and khorasan) to produce spontaneously fermented sourdough bread and to evaluate the impact on the dough rheological properties, ultrastructure and baking quality. Modern wheat sourdough bread and bakery yeast fermented bread were used as controls. Sourdoughs produced from modern and ancient wheats exerted different effects on dough viscoelastic properties, bread specific volume, texture, firming rate, colour and sensory properties, while there was no influence on bread water activity. Both khorasan sourdough, being characterised with the highest dough strength and dense gluten protein matrix, and emmer sourdough, with loose and thin gluten strands of low strength, yielded breads characterised by low specific volume and hard crumb texture. Spelt and modern wheat sourdough were characterised by foam-like dough structures with entrapped gas cells leading to breads of similar specific volume and texture. Although the yeast-fermented wheat flour exerted a higher specific volume and the lowest firmness, the sourdough wheat flour bread had a lower firming rate. A comparison of sourdough bread prepared with modern and ancient wheats revealed that breads based on ancient varieties possess a less noticeable sour taste, odour and flavour, thus contributing to more sensory-appealing sourdough bread.

Ancient Wheat Varieties and Sourdough Fermentation as a Tool to Increase Bioaccessibility of Phenolics and Antioxidant Capacity of Bread.

This study aimed to determine the impact of ancient wheat varieties (emmer, spelt and khorasan) and spontaneous sourdough fermentation on the bioaccessibility of total phenolic content (TPC) and the DPPH antioxidant capacity evolution during breadmaking and in vitro digestion. Sourdough and yeast-fermented modern wheat breads were used as controls. After 6 h of fermentation, the total titrable acidity of the sourdough increased from 139 to 167%. The wheat variety, type of fermentation and processing affected TPC, antioxidant activity and bioaccessibility. Antioxidant activity and TPC were reduced by dough mixing, increased after sourdough fermentation and slightly decreased or remained the same after baking. Although wheat flour had the highest TPC, the modeling of TPC kinetic revealed that emmer and spelt sourdough exhibited a higher bound phenolics release rate due to the higher acidity, which contributed to increased phenolics solubility. Although wheat bread, both before and after digestion, had the lowest TPC, especially the one prepared with yeast, high TPC bioaccessibilities and antioxidant activities after the digestion suggested that, except phenolics, digestion process improved the release of additional compounds with different bioaccessibility and biological activity. The results of this study proved that the application of sourdough fermentation can increase the potential of ancient wheats in the developing of functional bakery products.

Chia Seed Hydrogel as a Solid Fat Replacer and Structure Forming Agent in Gluten-Free Cookies.

Gluten-free cookies based on rice and chickpea flour with reduced-fat and increased protein content compared with conventional commercial gluten-free cookies were developed and used as a base for further vegetable fat replacement with chia seed hydrogel. Rheological properties of chia seed hydrogel revealed that 8% gels exhibited the optimal properties as a fat substitute. Designed cookie samples were characterized for their chemical composition, fatty acid profile, mineral content, physical, textural and color parameters, and sensory properties. All gluten-free cookies developed in this study could be labeled as "a source of iron and potassium", while those with chia seed hydrogel and cocoa powder could bear the additional claim "high in zinc and magnesium". Fat replacement with chia seed hydrogel resulted in a more favorable fatty acid composition with a PUFA/SFA ratio over 0.40 and nonsignificant changes in the cookies' hardness, weight, eccentricity, and specific volume, indicating that the chia seed hydrogel addition did not disturb the cookie structure and texture. The results of the sensory analysis confirmed that it is possible to apply chia seed hydrogel to produce reduced-fat cookies with sensory properties comparable to their full-fat counterpart and available commercial samples, and they are more appealing than commercial reduced-fat gluten-free cookies.

Effect of Sourdough and Whey Protein Addition on the Technological and Nutritive Characteristics of Sponge Cake.

Whey protein and sourdough ferment were used in different combinations to prepare functional sponge cakes, and their mutual influence on batter rheological behaviour as well as product physico-chemical, textural, colour and sensory properties were evaluated. All samples containing whey protein concentrate could bear the nutrition claim 'a source of protein'. The substitution of wheat flour with whey protein significantly influenced batter viscoelastic behaviour, lowered cake-specific volume, increased product hardness, chewiness, gumminess, and browning index and modified its sensory characteristics. The incorporation of sourdough in protein-enriched sponge cakes improved product-specific volume and appearance compared to a protein-containing sample without sourdough. Although sourdough addition has less of a deteriorating effect on sponge cake rheological and textural properties, when combined with whey protein, it led to a significant reduction in batter elasticity and an increase in product hardness. It was also shown that spontaneously fermented sourdough cannot act as the only leavening agent in sponge cake production. In general, the results of this study have shown that sourdough addition can contribute to improvement in protein-enriched sponge cake quality and that further investigations are necessary in terms of different sourdough and flour type incorporation to minimize the negative effects of protein addition.

Gluten-Free Crackers Based on Chickpea and Pumpkin Seed Press Cake Flour: Nutritional, Functional and Sensory Properties.

Research background: Despite the growing trend of the gluten-free market and the presence of a wide range of gluten-free products, there are still some shortcomings in nutritional and sensory quality of these products. The commercially available gluten-free products are characterised as products of inferior nutritional quality, particularly in terms of protein and dietary fibre content and with high glycaemic index. On the other hand, from a sensory point of view, gluten-free products usually have inappropriate textural and mechanical properties, poor mouthfeel and flavour. This is a consequence of the limiting choice of raw materials that mainly possess large amount of carbohydrate components. Experimental approach: Chickpea flour and two types of pumpkin seed press cake flour (virgin and cold pressed), at two substitution mass fractions (20 and 35%), were blended to produce gluten-free crackers without the presence of conventional gluten-free starch-rich ingredients. This study aims to investigate the effect of these non-conventional flours on nutritional and physicochemical properties, sensory acceptability, antioxidant activity and glycaemic index of crackers. Results and conclusions: All produced crackers can bear nutritional claims 'high fibre', 'source of protein' and 'source of minerals'. Replacing chickpea flour with pumpkin seed press cake flour increased protein and total phenolic content and enhanced antioxidant activity. The selected combination of raw materials allows the production of gluten-free crackers with a moderate glycaemic index. Besides nutrient content, the addition of cold-pressed flour increased overall sensory acceptability, noticeably improving taste and flavour scores compared to the control and crackers with virgin pumpkin seed flour. Novelty and scientific contribution: To the best of our knowledge, there is no study investigating the use of chickpea and pumpkin seed press cake flour blend without using conventional gluten-free flour and starch. The used non-conventional flour represents complementary raw materials in terms of protein quality and valuable alternatives to produce nutrient-rich, health-promoting gluten-free crackers with reduced glycaemic response and acceptable sensory properties.

A homogeneous split-luciferase assay for rapid and sensitive detection of anti-SARS CoV-2 antibodies.

Better diagnostic tools are needed to combat the ongoing COVID-19 pandemic. Here, to meet this urgent demand, we report a homogeneous immunoassay to detect IgG antibodies against SARS-CoV-2. This serological assay, called SATiN, is based on a tri-part Nanoluciferase (tNLuc) approach, in which the spike protein of SARS-CoV-2 and protein G, fused respectively to two different tNLuc tags, are used as antibody probes. Target engagement of the probes allows reconstitution of a functional luciferase in the presence of the third tNLuc component. The assay is performed directly in the liquid phase of patient sera and enables rapid, quantitative and low-cost detection. We show that SATiN has a similar sensitivity to ELISA, and its readouts are consistent with various neutralizing antibody assays. This proof-of-principle study suggests potential applications in diagnostics, as well as disease and vaccination management.

Vegetable By-Products as New Sources of Functional Proteins.

Vegetable by-products, obtained from cauliflower (CA), broccoli (BRL), cabbage (CB) and beetroot (BR) can be a potentially good source of proteins. The proteins were obtained from leaves (LPs) of vegetables with alkaline extraction at pH 10, and their isoelectric precipitation at pH 4. Protein contents were in the range of 39.76 - 53.33%, and the molecular weights of fractions were mostly about 45, 25 and 14 kDa. Their solubility is higher in the alkaline environment, where they reach the highest solubility at pH 10 (9.7 mg/mL for CALP, 8.45 for BRLP, 5.35 mg/mL for CBLP, 5.5 mg/mL for BELP). Moreover, they showed favorable emulsifying abilities, water absorption capacities (0.62 to 1.61 g/g) and foaming capacity (86.3 to 92%) as well as stability (48.57 to 79.30%). Digestibility was studied using gastrointestinal proteases (pepsin and pancreatin), and all four LPs can easily be digested. The biologically active potential of the digests was evaluated measuring antioxidant capacity by two complementary methods - DPPH+ and ABTS+ radical cation scavenging activity. The values for DPPH+ and ABTS+ were in the range from 59 to 65.1% at 0.1 and 0.3 mg/ml to 0.22 mg/ml IC50 values, respectively. Therefore, it can be indicated from these results, that obtained LPs, owing to their good functional properties, may be considered as potential ingredients of health-promoting food and cosmetic products.

Phytochemical assessment of plum (Prunus domestica L.) cultivars selected in Serbia.

The aim of this study was to assess content of primary and secondary metabolites of 18 plum cultivars selected in Serbia. Autochthonous cultivar 'Crvena Ranka' exhibited the highest level of total sugars (174.3 g/kg FW) while cultivar 'Mildora' had the highest sugars/acids ratio (17.2). Cultivar 'Nada' also contained high sugars/acid ratio as well as a high level of glucose and total sugars. Compared to the other cultivars, 'Nada' had a higher content of anthocyanins (27.14 mg/100 g FW), 'Crvena Ranka' neochlorogenic acid (23.21 mg/100 g FW) and flavanols (86.15 mg/100 g FW) but also the highest antioxidant activity. The PCA analysis confirmed that 'Nada' and 'Crvena Ranka' demonstrated better results in terms of sugars and antioxidant activity than the standard cultivars grown in Serbia and can be recommended for increased plum growing for fresh consumption, processing and in the breeding of plum with enhanced nutritional content.

Essential Gene Profiles for Human Pluripotent Stem Cells Identify Uncharacterized Genes and Substrate Dependencies.

Human pluripotent stem cells (hPSCs) provide an invaluable tool for modeling diseases and hold promise for regenerative medicine. For understanding pluripotency and lineage differentiation mechanisms, a critical first step involves systematically cataloging essential genes (EGs) that are indispensable for hPSC fitness, defined as cell reproduction in this study. To map essential genetic determinants of hPSC fitness, we performed genome-scale loss-of-function screens in an inducible Cas9 H1 hPSC line cultured on feeder cells and laminin to identify EGs. Among these, we found FOXH1 and VENTX, genes that encode transcription factors previously implicated in stem cell biology, as well as an uncharacterized gene, C22orf43/DRICH1. hPSC EGs are substantially different from other human model cell lines, and EGs in hPSCs are highly context dependent with respect to different growth substrates. Our CRISPR screens establish parameters for genome-wide screens in hPSCs, which will facilitate the characterization of unappreciated genetic regulators of hPSC biology.

Novel breads of non-wheat flours.

The aim of this study was to provide new approach in creating gluten-containing and gluten-free breads without additives by combining thermal and hydrothermal pretreatments of flours (rye, oat, sorghum and millet). The applied methodology included determinations of chemical composition of flours and breads, water absorption index, empirical and fundamental rheological measurements, and scanning electron microscopy, differential scanning calorimetry, colour, textural and sensory evaluations of breads. Novel rye, oat, sorghum and millet breads based on the blend of heat treated and extruded corresponding flours in ratio 70:30 were produced by conventional breadmaking process. All breads were characterized by increased fibre content and had appearance similar to common wheat bread. Gluten-free breads were harder, less elastic with more granular structure due to higher degree of starch crystallinity. Mixolab curves indicated on many possible ways for further breads optimisation.

Wheat breadmaking properties in dependance on wheat enzymes status and climate conditions.

The objective of this study was to evaluate albumins profile, proteolytic and amylolytic activity level and baking performance of wheat varieties grown in two production years with different climate conditions (2011 and 2012) in four locations. The results of ANOVA showed that variety, location, production year, and their interactions all had significant effects on all tested wheat quality parameters. The enzymatic activity and specific bread volume were mainly influenced by the variety. The samples from 2012 production year, had the lower values of albumin content, proteolytic and amylolytic activity, and bread specific volume. The correlation analysis, performed for 2011 production year, showed that albumin fraction (15-30 kDa) and proteolytic activity were negatively correlated with bread specific volume indicating the role of this fraction on lowering the crucial bread quality parameter. In 2012 production year, albumin fractions (5-15 kDa; 50-65 kDa) showed the most correlations, especially with parameters of bread quality.

Changes in the rheological properties of wheat dough during short-term storage of wheat.

BACKGROUND: The aim of the present study was to investigate the ability of the small- and large-deformation fundamental rheological tests to monitor the changes during short-term post-harvest storage of wheat. RESULTS: At the end of wheat storage period, wet gluten quantity decreased, while gluten index increased in comparison to those of freshly harvested samples. The rheological properties of dough changed over the pre-defined period of storage in terms of becoming more elastic and less extensible in comparison to that of freshly harvested wheat. Visco-elastic properties of weaker flour samples changed more markedly during storage than those of stronger flours, indicating that the intensity of dough rheological changes during wheat storage might be dependent on gluten quality and were the characteristic of wheat variety. CONCLUSION: It was shown that small deformation dynamic oscillation and large deformation creep-recovery tests can be successfully employed to monitor the changes in flour quality during wheat storage and that required storage period after wheat harvesting has to be defined according to wheat variety initial rheological properties and its gluten quality.

Content of free amino groups during postharvest wheat and flour maturation in relation to gluten quality.

The objective of this study was to monitor the changes in the content of free amino groups during postharvest wheat and flour maturation. The content of free amino groups of wheat flour was analysed immediately after wheat harvest, after 50 days of wheat storage and after 14 days of flour storage varying by wet gluten samples incubation temperatures and incubation times (0, 90 or 135 min at 30°C and after that 180 min at 37°C). The results were observed in relation to wheat-bug damaged kernels content, gluten index values, proteolytic activity and electrophoretic properties of gliadins and glutenins. The content of free amino groups increased during postharvest wheat and flour maturation periods. Proteolytic activity values were the highest 50 days after the wheat storage. The electrophoretic determination indicated a macromolecular redistribution of the gluten proteins from the moment of the wheat harvest until the moment of flour stabilisation.

Dystrophic calcifications and Raynaud's phenomenon in an eight-year old girl.

INTRODUCTION: Dystrophic calcifications are the most common subtype of skin calcinosis. Tumorous soft tissue calcium deposits usually contain hydroxyapatite and amorphous calcium phosphate. Differential diagnosis of skin calcinosis encompasses Thibierge-Weissenbach syndrome, systemic sclerosis, scleroderma, CREST syndrome (calcinosis, Raynaud's phenomenon, esophageal dysmotility, sclerodactyly and telangiectasia), dermatomyositis, systemic lupus erythematosus, ad myositis ossificans progressiva. CASE OUTLINE: We present the case of an eight-year old girl with tumorous soft tissue calcium deposits and Raynaud's phenomenon. At the age of 3.5 years, our patient was admitted to Pediatric Surgery Clinic because of bilateral acrocyanosis localized at the fingertips area of hands, with the signs of vascular trauma. Therapy with vasodilators and hyperbaric oxygen treatment were completed. This therapy resulted in improvement. At the age of eight, the patient was admitted again due to intermittent, painful cramps localized in both hands. Punctiform deposits were present at the tips of fingers and toes, which looked like calcifications and were spontaneously eliminated, with the remnants of crater-shaped defects. A hard tumorous deformity localized in soft tissue was present in the extensor area of the right elbow. Laboratory indicators of inflammation were within the reference values, and antinuclear antibodies were positive. A nodus localized at the right elbow was extirpated. Pathohistological findings: connective and fat tissue with large deposits of calcium. CONCLUSION: Further follow-up of our patient is necessary due to possible development of complete picture of CREST syndrome or systemic sclerosis.

Determination of free sulphydryl groups in wheat gluten under the influence of different time and temperature of incubation: method validation.

The aim of the present study was to determine the characteristics of an analytical method for determination of free sulphydryl (SH) groups of wheat gluten performed with previous gluten incubation for variable times (45, 90 and 135min) at variable temperatures (30 and 37°C), in order to determine its fitness-for-purpose. It was observed that the increase in temperature and gluten incubation time caused the increase in the amount of free SH groups, with more dynamic changes at 37°C. The method characteristics identified as relevant were: linearity, limit of detection, limit of quantification, precision (repeatability and reproducibility) and measurement uncertainty, which were checked within the validation protocol, while the method performance was monitored by X- and R-control charts. Identified method characteristics demonstrated its acceptable fitness-for-purpose, when assay included previous gluten incubation at 30°C. Although the method repeatability at 37°C was acceptable, the corresponding reproducibility did not meet the performance criterion on the basis of HORRAT value (HORRAT<2).

Resveratrol has anti-leukemic activity associated with decreased O-GlcNAcylated proteins.

CLL cells are characterized by high levels of proteins that are post-translationally modified by O-linked ß-N-acetylglucosamine (O-GlcNAc) moieties, but it is not clear whether O-GlcNAc is a relevant therapeutic target. The neutraceutical resveratrol is cytotoxic to chronic lymphocytic leukemia cells in vitro. In this study, we found that resveratrol has therapeutic activity as a single agent in vivo in both human chronic lymphocytic leukemia patients and mice with erythroleukemia. Blood and splenic O-GlcNAc levels reflected the changes in tumor burden. Resveratrol directly lowered O-GlcNAc levels in leukemia cells through proteasomal activation, but increasing O-GlcNAc levels in vitro did not prevent cell death. These findings suggest that resveratrol has potential as a novel treatment for some forms of chronic and acute leukemia, and the measurement of O-GlcNAc levels could be a surrogate marker for therapeutic responses.

Recurrent infections and cows-milk hypersensitivity in a 2-year-old girl with hyper immunoglobulin E syndrome.

The hyperimmunoglobulin E syndrome is a rare complex primary immunodeficiency characterized by high serum IgE levels, eczema, and recurrent infections. We present a case of 2-years-old girl with eczema and repeated bacterial skin and lung infections since the period of infancy. The patient also had eosinophilia, high serum levels of IgE, and cows-milk hypersensitivity. We describe the case, in order to illustrate the difficulty of establishing the diagnosis hyper-immunoglobulin E syndrome in a toddler.

Aberrant interferon-signaling is associated with aggressive chronic lymphocytic leukemia.

The type I interferons (IFNs) normally suppress tumor growth by phosphorylating and activating the signal transducer and activator of transcription 1 (STAT1), but also briefly activate STAT3, which promotes cell growth. In chronic lymphocytic leukemia (CLL) cells, the duration of IFN-mediated STAT3 phosphorylation was found to exhibit significant interpatient variability and was prolonged in cells with high risk features, such as 11q- and 17p-deletions involving ataxia telangiectasia mutated (ATM) and p53. This aberrant signaling pattern was associated with a paradoxical increase in cell size and number in response to IFN and similar alterations in IFN-signaling and responses were seen in cell lines that developed in the absence of p53 or ATM. However, direct inhibition of p53 or ATM failed to cause these changes, and CLL cells with aggressive clinical features were found to also express high levels of reactive oxygen species (ROS), which decrease tyrosine phosphatase activity. Prolonged IFN-mediated STAT3 phosphorylation and lowered phosphatase activity could be reversed by antioxidants. These findings suggest that increased ROS levels may corrupt IFN-signaling processes in aggressive CLL cells, causing IFN to be used as a growth factor rather than a tumor suppressor. Antioxidants or STAT3 kinase inhibitors might improve the outcome of IFN therapy in CLL by restoring normal signaling.

[Possibilities of the primary and secondary prophylaxis in treatment of childhood asthma].

INTRODUCTION: The natural course of asthma is unpredictable and appears to be unaffected by any therapeutic strategy. Under such circumstances, the attention must be focused on the opportunities for prevention of a disease which is chronic, life long and incurable, even thought it can be very effectively controlled. During the past decades, a lot of a studies have been performed and started, in which relatively large numbers of children were included and followed prospectively to determine the incidence of risk factors for asthma in childhood. All these studies have contributed significant new information. The levels of prevention must be considered in all patients. There are two main separate components to the strategy. PRIMARY PROPHYLAXIS: Primary prophylaxis (time course of allergic sensitization, timing of exposure to allergens, influence of tobacco smoke, maternal health and allergen exposure) is introduced before there is any evidence of sensitization to factors which might have caused the disease. There is increasing evidence that allergic sensitization is a very common precursor to the development of asthma. SECONDARY PROPHYLAXIS: Secondary prophylaxis (allergen avoidance, hygiene hypothesis) is important after primary sensitization to allergen has occurred, but before there is any evidence of asthma. CONCLUSION: In this article the authors reviewed all results of studies about primary and secondary prophylaxis of asthma and its influence on the course of disease.