Addition Of Sugar Research Articles

Fried Chocolate Bread with Panir Flour untuk Tumbuh Kembang Anak Pada Pondok Pesantren Wihdatul Ulum Di Desa Bontokassi Kecamatan Parangloe Kabupaten Gowa

Indonesia has a large population, especially children. Chocolate is almost liked by all groups. In addition to being delicious, chocolate also has various benefits forthe body, from improving mood to maintaining heart health. Starting from children, teenagers, to adults, many like to eat chocolate. In addition to its distinctive and delicious taste, chocolate also has health benefits. Chocolate is a processed cocoa bean that is roasted, ground, then formed into a liquid, paste, or in block form. Dark chocolate or dark chocolate contains antioxidants, vitamins, and minerals that are good for the body. Now there are many chocolates that are processed with the addition of sugar, milk, and so on. Instead of getting the health benefits of chocolate, consuming this type of chocolate is dangerous for health. Therefore, if you want to feel the benefits of chocolate, consume dark chocolate without adding excess sugar. because this product has an affordable price and has great benefits.

Inhibition of ketohexokinase prevents fructose-induced insulin resistance and endothelial dysfunction in high fat and sucrose-fed mice via enhancing energy expenditure

Abstract Introduction Cardiovascular disease linked to fatty liver poses a significant threat to health. Consumption of high-fat diets (HFD) is known to trigger a cascade of health issues including obesity, diabetes and non-alcoholic fatty liver disease (NAFLD). These conditions are exacerbated, with the addition of dietary sugar to high-fat diets (HFSD), resulting in non-alcoholic steatohepatitis (NASH). Studies have shown that fructokinase (ketohexokinase or KHK) knockdown protects mice from HFSD-induced NASH. Purpose This study focused on examining the temporal changes of metabolic perturbations, triggered by sugar consumption, particularly in the transition from fatty liver to cardiovascular disease. Methods C57/BL6 wild type (WT) and KHK-knockout mice were fed with HFD or HFSD for different durations between 6 to 20 weeks, to induce NAFLD and NASH. Western blotting and quantitative-PCR were carried out on tissues and isolated endothelial cells for protein and mRNA expressions, respectively. Glucose and insulin tolerance tests and whole-body energy measurements were performed at different time-points. Combined Laboratory Animal Monitoring System was used to monitor various parameters of energy expenditure. Functional interactions between endothelial cells and adipose tissues were carried out using an in-house Organ-On-a-chip technology. Endothelial function was studied by measuring isometric tensions in organ bath. Results KHK-deficient mice had significantly lower weight compared to WT littermates. Wild-type mice fed HFD and HFSD exhibited significant glucose intolerance, insulin resistance, and endothelial dysfunction compared to KHK-knockout mice. However, HFSD led to more severe outcomes compared to HFD, demonstrating differing temporal effects on glucose intolerance and insulin resistance. Interestingly, KHK-knockout mice were significantly protected from the effects induced by HFSD, as well as, such as increased body weight, fasting hyperglycemia, hyperinsulinemia and endothelial function. KHK-knockout mice exhibited significant increase in oxygen consumption and energy expenditure, both in HFD and HFSD-fed conditions. These data suggest that high energy-expenditure is one of the primary mechanisms, in reducing adiposity and body weight in HFSD-fed knockout mice. Notably, KHK-deficient mice were protected against sugar-induced fat accumulation, random hyperinsulinemia and significant impairment of endothelial function. Conclusions A novel finding in this study is that KHK ablation protects not only against HFSD-induced, but also HFD-induced pathologies via significantly enhancing energy expenditure. The results suggest that targeting KHK could potentially mitigate the development of fatty liver and cardiovascular diseases, induced by high calorie diets.

Synchronous Modulation of H-bond Interaction and Steric Hindrance via Bio-molecular Additive Screening in Zn Batteries.

In addressing challenges such as side reaction and dendrite formation, electrolyte modification with bio-molecule sugar species has emerged as a promising avenue for Zn anode stabilization. Nevertheless, considering the structural variability of sugar, a comprehensive screening strategy is meaningful yet remains elusive. Herein, we report the usage of sugar additives as a representative of bio-molecules to develop a screening descriptor based on the modulation of the hydrogen bond component and electron transfer kinetics. It is found that xylo-oligosaccharide (Xos) with the highest H-bond acceptor ratio enables efficient water binding, affording stable Zn/electrolyte interphase to alleviate hydrogen evolution. Meanwhile, sluggish reduction originated from the steric hindrance of Xos contributes to optimized Zn deposition. With such a selected additive in hand, the Zn||ZnVO full cells demonstrate durable operation. This study is anticipated to provide a rational guidance in sugar additive selection for aqueous Zn batteries.

The pH and Sucrose Influence Rhamnolipid Action Toward Planktonic and Biofilms of Listeria monocytogenes.

Bacterial resistance and persistence in food environments are major concerns for the industry, which constantly seeks new strategies to reduce microbial contamination. Rhamnolipids (RL) biosurfactants are considered sustainable and green alternatives to synthetics; furthermore, they have demonstrated potential for controlling various foodborne pathogens. Food environments are typically exposed to diverse pH, solutes, temperatures, and water activity (aw) levels that may favor the survival of pathogens. Therefore, it is crucial to consider these factors in evaluating the performance of novel antimicrobials. Our study examined the influence of pH and sucrose on the antimicrobial activity of RL against both planktonic and biofilm of Listeria monocytogenes. We found that the presence of sucrose can enhance the antimicrobial effectiveness of RL against both planktonic and sessile bacteria. The addition of sugar particularly improved RL action at pH 6 and 7. Moreover, we observed that the type and size of RL self-assembly structures depend on the pH and sucrose concentration. These findings suggest potential for developing RL-based innovative methods to control L. monocytogenes in sugar-rich or -low aw foods and environments.

Cellular scaffolding: Crowdsourcing cellular responses in health and disease

Cellular scaffolding: Crowdsourcing cellular responses in health and disease In this article, Dr Alan Herbert discusses how different types of cellular scaffolds interact and impact the risk of diseases, citing the example of Z-RNAs pushing cells to inflammatory states in tumors and autoimmune conditions, setting the stage for new therapeutics. The cell is full of so many proteins and RNA versions that it is a challenge to see how anything works at all. There is so much crowding it is like trying to get to your train through the mass of people on the station platform during rush hour. All the different biomolecules involved can be modified by the addition of phosphates, sugars, and lipids in so many different ways and at so many alternative sites that it seems impossible to track the one that matters. Yet, somehow, cells work in a highly efficient manner.

An innovative technology for producing agricultural Jiaosu from fruit waste without additional sugar and water: Fermentation characteristics, microbial community, and its impact on the growth of pakchoi

Landfilling or incineration of fruit waste causes serious environmental problems and economic losses. Agricultural Jiaosu (AJ) can convert fruit waste into agricultural inputs. However, traditional AJ preparation requires large amounts of brown sugar and water, restricting its large-scale application. In this study, five types of single fruit waste and one type of mixed fruit waste were used to produce AJ without additional of sugar and water. The physicochemical properties and microbial community dynamics of AJ fermentation were analyzed, and the effects and potential mechanism of AJ in promoting pakchoi growth were evaluated. The results showed that during the digestion process, the pH value of all AJ gradually decreased to below 3.6. Organic acids were the main metabolites, lactic acid and acetic acid accounted for 46.55 %–99.22 % of all organic acids. As fermentation progressed, Firmicutes gradually replaced Proteobacteria as the most dominant phylum, accounting for 54.1 %–99.41 % of the microbial communities. After 90 days, Lactobacillus and Lentilactobacillus were the dominant genera in all AJ, accounting for 44.49 %–92.54 % of the communities. Structural equation modeling revealed that AJ promoted plant growth through organic acids and bacterial community. This study is the first time to propose the concept of producing AJ from fruit waste without additional sugar and water and reveals the fermentation characteristics and microbial dynamics of AJ, as well as its potential mechanism as a biological inoculant in promoting plant growth. This study provides a new approach for the large-scale utilization of fruit waste to produce green agricultural inputs.Abbreviations: AJ, agricultural Jiaosu; ANOVA, analysis of variance; ASVs, amplicon sequence variants; DNA, deoxyribonucleic acid; HPLC, high performance liquid chromatography; LDA, linear discriminant analysis; LEfSe, linear discriminant analysis effect size; PCoA, Principal coordinates analysis; PCR, polymerase chain reaction; RDA, redundancy analysis; rRNA, ribosomal RNA; SEM, structural equation modeling.

Characterization of Robusta Coffee Powder with Addition of Palm Sugar and Vanilla Powder to Improve The Quality of Robusta Coffee (Coffea canephora Pierre)

Bondowoso Robusta Coffee is a type of Robusta coffee that has a distinctive taste and high quality. The distinctive taste and high quality are obtained from the results of harvest management and post-harvest processing in accordance with Standard Operating Procedures (SOP). This research will be carried out by testing the characteristics of Robusta ground coffee with a mixture of palm sugar and vanilla flavoring to improve the quality of Robusta ground coffee. This was done by knowing the physicochemical and organoleptic properties of the mixed Robusta ground coffee. The aim of this research is to determine the interaction between roasting level and the composition of palm sugar and vanilla powder on the physiochemical and organoleptic properties of 3 in 1 Robusta Coffee. This research uses an experimental plan arranged factorially with the basic pattern of a Completely Randomized Design (CRD) with 4 replications. This design has two factors, the first factor is the roasting level and the second is the mixture composition. The results of this research show (1) The interaction of Roasting Level and Mixture (palm sugar and vanilla powder) has no significant effect on all observed variables such as water content, Brix content, pH value, and powder bulk density. (2) The effect of the main level of roasting has a very significant effect on the observed variables of rainfall density and an insignificant effect on the variables of air content, Brix content and pH value. (3) The effect of the main mixture has a very significant effect on the observed variables of air content, Brix content and powder bulk density and has no significant effect on the variable pH value.

ALTERNATIVE NATURAL SWEETENERS AS SUGAR SUBSTITUTES USED IN MAKING ICE CREAM

Ice cream is a frozen dessert highly appreciated by all categories of consumers. Lately, consumers' attention is focused on healthy products, with as few ingredients as possible or with natural ingredients. By definition, ice cream is a sweet product. As a rule, sugar is used to obtain ice cream for its characteristics (sweet taste, creaminess, texture, melting time), but the new requirements require the addition or substitution of sugar with other sweeteners. Consumers still prefer the sweet taste of artificial sweeteners, but they are cultivating their taste and increasingly choosing natural nutritious or non-nutritive sweeteners as consumer awareness of healthy foods has increased. This manuscript was based on the study of the literature regarding the use of natural sweeteners in ice cream formulations.

Consumption of energy drinks among medical university students in Kazakhstan.

Due to lack of information and deceptive advertising, young people widely consume energy drinks, although they should be careful because of their contents. Energy drinks contain high amounts of caffeine, sugar, and other additives that can stimulate the body. They can cause irritability, anxiety, insomnia and can increase heart rate, blood pressure and respiratory rate. Aim of this research was to study consumption of energy drinks among medical university students in Kazakhstan. Cross-sectional study was conducted. A web-based questionnaire was used to collect information. Students enrolled in various courses at the medical university, including interns and residents, were invited to participate. Atotal of 764 respondents took part in the survey. Among the trainees, 41.4 % had consumed caffeinated energy drinks in the past six months, and at the time of the survey, 16.2 % of the respondents had consumed these drinks. Respondents who do not drink alcohol are 3 times (p<0.001) more likely not to use energy drinks compared to those who drink alcohol. Those who do not exercise are 4.7 times (p<0.001) more likely not to consume energy drinks compared to active athletes. Participants who had not consumed caffeinated energy drinks in the past six months were 1.8 times (p<0.001) more likely not to consume energy drinks compared to those who drank them. Consumption of energy drinks by students can temporarily increase physical and mental performance, improve mood and raise energy levels, while at the same time, they also come with serious health risks. Therefore, educating students about the potential risks and the importance of a healthy lifestyle is essential.

Sugar taxation policy and sugar consumption in Malaysia

Malaysia has been taking initiatives to combat obesity as well as non-communicable diseases. A sugar-sweetened beverage (SSBs) tax was proposed in 2014 and implemented in 2019 with an excise duty of RM0.40 per litre for ready-to-drink sugar content. This article aimed to spark a discussion about Malaysia's current sugar tax policies to reduce excessive sugar consumption among Malaysians. However, the alarming rise in the number of obesity and diet-related noncommunicable diseases indicates that the sugar tax control measures are less effective in combating health issues. This study compares Malaysia's neighbours' sugar tax policies and other sugar consumption-related measures. To combat the emerging sugar-related health issues in Malaysia, the government should collaborate with food manufacturers to reduce the amount of sugar in their food production, abate sugar-containing products’ promotion, educate the consequences of excess sugar consumption at all levels of education, apply sugar taxes with lower tax base rate and specific tax rate depends on sugar addition, and form a regional policy between neighbours.

Testing AI predictions that hyperglycemia-induced non-enzymatic glycation of immune system proteins impairs diabetic immunity

People with type 1, type 2 and gestational diabetes, and newborns of diabetic mothers, have increased risks of contracting microbial infections such as SARS-CoV-2, amplifying comorbidity and mortality risks. Hyperglycemia and its associated hyperinflammation mediate these risks but by unknown mechanisms. We hypothesize that hyperglycemia results in glycation (non-enzymatic addition of sugars to free amino groups such as lysines) of antimicrobial and anti-inflammatory proteins and peptides that mediate innate and adaptive immunity. While glycation of hemoglobin A1c is used as a benchmark for glycemic control, studies of the effects of the glycation of immune system molecules are almost non-existent. Three AI protein-glycation prediction models demonstrate that immune system proteins and peptides such as serotransferrin, lactotransferrin, lysozyme, complement proteins, immunoglobulins, Toll-like receptors, etc., probably glycate at multiple sites affecting function. However, the AI-predicted glycation sites differed significantly among the three models employed. Therefore, three tests of these predictions were carried out. Predicted glycation sites were first evaluated in terms of clinically or experimentally identified glycation sites of homologous proteins and known effects of lysine substitutions at predicted glycation sites. Second, mass spectrometry was used to identify glycation sites on lactoferrin and human lysozyme. Third, the effects on the antimicrobial activity of lactotransferrin and lysozyme glycation were tested using a Candida albicans assay. Results of tests demonstrate that protein glycation occurs and interferes with immune function but that AI glycation prediction models are very inaccurate. Suggestions for improving AI accuracy are provided.

Low‐field nuclear magnetic resonance relaxometry: a new approach for monosaccharide identification in sugar solutions

SummaryMonosaccharides such as glucose, fructose, and galactose are the building blocks of oligosaccharides and act as the major energy sources in our body. These three monosaccharides are the most prominent in food processing and widely consumed analytes, found in fruits or milk. Although several analytical methods are available to identify and quantify sugar solutions, their drawbacks urge the search for better possible alternatives. A simple, rapid, and efficient method for identifying monosaccharides (ᴅ‐fructose, ᴅ‐galactose, and ᴅ‐glucose) was investigated using low‐field nuclear magnetic resonance (LF‐NMR), exploring the applicability and reliability of this machine. The transverse (T2) relaxation curve was analysed to distinguish monosaccharides from each other. The increasing monosaccharide concentration in the sugar solution causes leftward shifts in signals, indicating a gradual reduction in the mobility of water molecules. The addition of sugar generated a secondary peak, which assisted in identifying monosaccharides. Notably, fructose exhibited distinct behaviour from that of glucose and galactose. The regression coefficient consistently exceeded 0.98, indicating the reliability of this experiment. LF‐NMR encountered challenges in differentiating stereo‐isomeric glucose and galactose. However, quantification of monosaccharides from known binary mixtures (water and one monosaccharide) is possible by preparing standard curves of different concentrations of sugars. These standard curves can be either T2 or the total surface area of the peaks as a function of sugar concentration. The study concludes that LF‐NMR has potential for the qualitative and quantitative analysis of monosaccharides in pure solutions or powdered samples. It is hoped that the rapid and easy way of monosaccharide identification by simple analysis of peaks obtained from NMR spectra will enhance its accessibility for users. However, it also acknowledges limitations in applying this method to complex systems (mixtures of different sugars in a solution).

Potensi Minuman Probiotik Berbahan Dasar Campuran Buah (Jambu, Lemon, Melon, Bit) Sebagai Minuman Kesehatan

Background; The health of the digestive tract is very important for maintaining the body's immune system, in maintaining the health of the digestive tract there are two components of probiotics and prebiotics are needed. Probiotics are good bacteria that are beneficial for the health of the digestive tract. Probiotics can be obtained through food or drink that has gone through a fermentation process using gram-positive bacteria. Probiotics produce bacteriocins and several compounds needed by the body such as lactic acid, acetic acid, hydrogen peroxide, lactoperoxidase and lipoperoxide, pyridoxine, niacin, folic acid, cobalamin and biotin which are useful for boosting the body's immune system. Objectives; The purpose of this study was to make a probiotic drink with fruit-based ingredients (guava, lemon, melon, beet) as a probiotic product that can be an alternative health drink. Material and Method; This study used an experimental design with a completely randomized design, where there were three formulas that had differences in the addition of sugar (F1: 10% sugar, F2: 20% and F3: 30%). The materials used in this study included: Guava, Lemon, Melon,Beets, Sugar and used the Lactobacillus plantarum bacterial starter. The tests carried out included: organoleptic test, total plate count test and acidity test Results; the organoleptic test results, it was found that Formula 3 was the most preferred with organoleptic qualities: slightly sour, slightly muddy and very red in color. The red color comes from beets. The acidity level of each formula includes: F1 (3.43), F2 (3.39), F3 (3.34). The results of the total plate number test showed that the average total plate number of the product was 1.7 x 10 8/CFU/ml.Keywords: Beets, Guava, Healthy drink, Lemon, Melon, Probiotics

High Sugar and Fat Additives in Fast Food Restaurants: Is Whether Addiction or Consumer Loyalty?

The article will describe how chemicals in fast food and consumer loyalty to consume in high frequency. The chemical reality of fast food is suspected to cause addiction so that on the other hand consumers will appear loyal. This research applies a systematic review to examine studies exploring customer loyalty in the context of fast-food restaurants. The systematic review was chosen as the most appropriate method because its aim is to collect, critically evaluate, integrate, and present findings from various research studies relevant to the research question or topic. The findings offer significant theoretical implications for future research in this field and practical implications for practitioners in the fast-food industry. The initial relevance of each manuscript was determined based on its title. Manuscripts that appeared to discuss customer loyalty within the fast-food industry were selected for further evaluation. This study collected comprehensive information from these selected studies, including the author, year, title, and abstract. It’s noteworthy that all references utilized in this study are derived exclusively from English-language literature. The research contributes to the existing literature by providing a comprehensive and updated summary of the antecedents and their effects, and it offers actionable insights and recommendations on how to enhance customer loyalty, thereby providing a competitive advantage in the fast-food industry.

Ultra-Processed Food Consumption and Its Role in Obesity and Metabolic Syndrome Development

This article examined the relationship between the consumption of ultra-processed foods (UPF) and obesity, as well as metabolic syndrome, through a systematic review approach. UPFs were known to contain salt, sugar, oils, fats, and other additives mimicking natural foods' sensory qualities. Research indicated that diets high in UPF were nutritionally unbalanced and promoted excessive consumption, which was detrimental to health. Data from various studies showed that increased UPF consumption was linked to a higher risk of obesity and related diseases, such as type 2 diabetes. Cross-sectional studies indicated that UPF consumption correlated with a higher risk of overweight in men compared to women. This research utilized diverse data sources that used methods such as semi-quantitative food frequency questionnaires and prospective analysis to evaluate this relationship. Results showed that high UPF consumption significantly contributed to obesity and metabolic disorders, affecting metabolic profiles and overall diet quality, highlighting the need for better dietary policies to mitigate the adverse impacts of ultra-processed foods.

Process Optimization for Antarctic Krill (Euphausia superba) Sauce Based on Back Propagation Neural Network Combined with Genetic Algorithm

Using Antarctic krill (Euphausia superba) as the research object, we optimized the process conditions for Antarctic krill sauce (AkS) by including three factors (salt addition, rock sugar addition, and the oil-to-material ratio) and sensory evaluation as response values. The data from the response surface were fed into the back propagation (BP) neural network training, generating a model mapping the process conditions and sensory scores, which were subsequently combined with the genetic algorithm (GA) for global optimization to determine the optimal process for AkS preparation. The results revealed that the response surface model was well suited to the BP neural network training and prediction sets, with correlation values of 0.98 and 0.95, respectively. The fitting prediction effect was obvious for the sensory scoring results of the product. The parameters obtained from the GA’s global optimization search accord with the analytical results of the response surface. The findings demonstrated that combining a BP neural network with a GA can enhance the AkS preparation technique. Under optimal processing conditions, AkS has a high sensory score and protein and carbohydrate contents, moderate fat content, minimal fat oxidation, and non-detectable pathogens, indicating that the AkS in this study was nutritious and safe to consume.

Preparation of Novel Marine Enterococcus faecium MSD8 Exopolysaccharide Ointment and In Vivo Evaluation of Its Impact on Cutaneous Wound Healing in Male Albino Rats.

The current study describes the isolation of exopolysaccharide (EPS) producing lactic acid bacteria (LAB) from marine samples and testing different sugar additives with different proportions for enhanced EPS yield. The isolate MSD8 showed the most potential, yielding 200 mg/L of EPS after being cultivated at 37 °C for48 h on de Man Rogosa and Sharpe medium (MRS) supplemented with3% sucrose. The marine isolate MSD8 was identified as Enterococcus faecium with 99.58% probability using 16S rRNA gene sequencing. The obtained sequence was deposited in GenBank and assignedthe accession number MW924065. The feature of MSD8-EPS was characterized by estimating the total carbohydrate content by UV-vis to be ~ 71%. The FTIR analysis further indicated the presence of characteristic bands of polysaccharide. The cytotoxicity of the produced MSD8-EPS was assessed using human skin fibroblasts (HSF). The IC50 was determined to be > 100 μg/mL, which signifies that MSD8-EPS is safe for skin application. The produced EPS was used to prepare a novel ointment, which was tested for wound healing ability in male albino rats. The ointment significantly (P ≤ 0.05) shortened the time needed for wound healing, as it successfully healed the wounds by 94.93% on the 7th day and completely (100%) healed the wound by the 12th day. In comparison, the control group was healed by 73.2% and 84.83%, respectively. The data confirm that the prepared ointment can safely be used for pharmaceutical wound care products.

Effect of sugar or sugar alcohol with different spatial structures on the stability of paclitaxel/HS15/Tween80 micelles: Based on micellization thermodynamics

Paclitaxel (PTX) is clinically recommended for the treatment of breast, ovarian and non-small cell lung cancer. This study delves into the impact of sugars (mannose or glucose) and sugar alcohols (mannitol or sorbitol) with varying spatial structures on the interfacial and thermodynamic behavior of the PTX/HS15/Tween80 micellar system. The research reveals that the presence of OH groups in sugars and sugar alcohols enhances hydrogen bonding force between surfactant and water, altering the critical micelle concentration (CMC) in solution and the surfactant’s arrangement at the interface, thus facilitating micelle formation and enhancing system stability. Dynamic light scattering (DLS) results show that the particle size of the PTX/HS15/Tween80 system is 11.69 nm, with micellar size showing a positive correlation with additive concentration. Sugar alcohols exhibit a more pronounced effect on increasing micelle size compared to sugars. Molecules featuring outward-facing OH groups demonstrating a greater ability to facilitate the formation of smaller micelle than those with inward-facing groups, emphasizing the influence of spatial structural differences. Stability experiments demonstrate that light transmission remains above 97 % in the system for 8 h after the addition of sugar, and the relative absorbance of PTX is maintained above 97 % within 6 h. Results from FD-B21 fluorescence probe release further confirm the positive impact of sugars and sugar alcohols on the stability of PTX/HS15/Tween80 micelles, highlighting stability differences based on molecules with distinct spatial structures. When OH groups on sugar or sugar alcohol molecules are more oriented in the same direction, intramolecular hydrogen bonds are formed, reducing spatial resistance and promoting the formation of micellar with smaller particle sizes. This process enhances physical stability by reducing inter-molecular collisions. In summary, the presence of sugars and sugar alcohols promotes micelle formation and boosts system stability, offering valuable insights for enhancing the stability of drug-loaded micelles.

In vivo mapping of the mouse Galnt3-specific O-glycoproteome

The UDP-N-acetylgalactosamine polypeptide:N-acetylgalactosaminyltransferase (GalNAc-T) family of enzymes initiates O-linked glycosylation by catalyzing the addition of the first GalNAc sugar to serine or threonine on proteins destined to be membrane-bound or secreted. Defects in individual isoforms of the GalNAc-T family can lead to certain congenital disorders of glycosylation (CDG). The GALNT3-CDG, is caused by mutations in GALNT3, resulting in hyperphosphatemic familial tumoral calcinosis (HFTC) due to impaired glycosylation of the phosphate-regulating hormone FGF23 within osteocytes of the bone. Patients with hyperphosphatemia present altered bone density, abnormal tooth structure and calcified masses throughout the body. It is therefore important to identify all potential substrates of GalNAc-T3 throughout the body to understand the complex disease phenotypes. Here, we compared the Galnt3-/- mouse model, which partially phenocopies GALNT3-CDG, with wild-type mice and employed a multi-component approach utilizing chemoenzymatic conditions, a product-dependent method constructed using EThcD triggered scans in a mass spectrometry workflow, quantitative O-glycoproteomics, and global proteomics to identify 663 Galnt3-specific O-glycosites from 269 glycoproteins across multiple tissues. Consistent with the mouse and human phenotypes, functional networks of glycoproteins that contain GalNAc-T3-specific O-glycosites involved in skeletal morphology, mineral level maintenance and hemostasis were identified. This library of in vivo GalNAc-T3-specific substrate proteins and O-glycosites will serve as a valuable resource to understand the functional implications of O-glycosylation and to unravel the underlying causes of complex human GALNT3-CDG phenotypes.

Plant production of recombinant antigens containing the receptor binding domain (RBD) of two SARS-CoV-2 variants.

The aim of this work was to rapidly produce in plats two recombinant antigens (RBDw-Fc and RBDo-Fc) containing the receptor binding domain (RBD) of the spike (S) protein from SARS-CoV-2 variants Wuhan and Omicron as fusion proteins to the Fc portion of a murine IgG2a antibody constant region (Fc). The two recombinant antigens were expressed in Nicotiana benthamiana plants, engineered to avoid the addition of N-linked plant-typical sugars, through vacuum agroinfiltration and showed comparable purification yields (about 35mg/kg leaf fresh weight). Their Western blotting and Coomassie staining evidenced the occurrence of major in planta proteolysis in the region between the RBD and Fc, which was particularly evident in RBDw-Fc, the only antigen bearing the HRV 3C cysteine protease recognition site. The two RBD N-linked glycosylation sites showed very homogeneous profiles free from plant-typical sugars, with the most abundant glycoform represented by the complex sugar GlcNAc4Man3. Both antigens were specifically recognised in Western Blot analysis by the anti-SARS-CoV-2 human neutralizing monoclonal antibody J08-MUT and RBDw-Fc was successfully used in competitive ELISA experiments for binding to the angiotensin-converting enzyme 2 receptor to verify the neutralizing capacity of the serum from vaccinated patients. Both SARS-Cov-2 antigens fused to a murine Fc region were rapidly and functionally produced in plants with potential applications in diagnostics.