PASTA-kinase-mediated signaling drives accumulation of the peptidoglycan synthesis protein MurAA to promote cephalosporin resistance in Enterococcus faecalis.

The bacterial PASTA kinase, IreK, is required for intrinsic cephalosporin resistance in the Gram-positive opportunistic pathogen, Enterococcus faecalis. IreK activity is enhanced in response to cell wall stress, such as cephalosporin exposure. The downstream consequences of IreK activation are not well understood in E. faecalis, but recent work in other low-GC Gram-positive bacteria demonstrated PASTA kinase-dependent regulation of MurAA, an enzyme that performs the first committed step in the peptidoglycan synthesis pathway. Here, we used genetic suppressor selections to identify MurAA as a downstream target of IreK signaling in E. faecalis. Using complementary genetic and biochemical approaches, we demonstrated that MurAA abundance is regulated by IreK signaling in response to physiologically relevant cell wall stress to modulate substrate flux through the peptidoglycan synthesis pathway. Specifically, the IreK substrate, IreB, promotes proteolysis of MurAA through a direct physical interaction in a manner responsive to phosphorylation by IreK. MurAB, a homolog of MurAA, also promotes MurAA proteolysis and interacts directly with IreB. Our results therefore establish a connection between the cell wall stress sensor IreK and one critical physiological output to modulate peptidoglycan synthesis and drive cephalosporin resistance.

Secrets of getting started: Regulation of the first committed step of peptidoglycan synthesis by protein phosphorylation in Enterococcus and other Gram-positive bacteria.

Regulation of the first committed step of peptidoglycan precursor synthesis by MurA-enzyme homologs has recently taken center stage in many different bacteria. In different low-GC Gram-positive bacteria, regulation of this step has been shown to be regulated by phosphorylation of homologs of the IreB/ReoM regulatory protein by PASTA-domain Ser/Thr-protein kinases. In this issue, Mascari, Little, and Kristich determine this regulatory pathway and its links to resistance to cephalosporin ß-lactam antibiotics in the major human pathogen, Enterococcus faecalis (Efa). Unbiased genetic selections identified MurAA (MurA-family homolog) as the downstream target of IreB regulation in the absence of the IreK Ser/Thr-protein kinase. Physiological and biochemical approaches, including determination of MICs to ceftriaxone, Western blotting of MurAA cellular amounts, isotope incorporation into peptidoglycan sacculi, and thermal-shift binding assays of purified proteins, demonstrated that unphosphorylated IreB, together with proteins MurAB (MurZ-family homolog), and ReoY(Efa) negatively regulate MurAA stability and cellular amount by the ClpCP protease. Importantly, this paper supports the idea that ceftriaxone stimulates phosphorylation of IreB, which leads to increased cellular MurAA amount and precursor pathway flux required for E. faecalis cephalosporin resistance. Overall, findings in this paper significantly contribute to understanding variations of this central regulatory pathway in other low-GC Gram-positive bacteria.

Nutritional composition, lipid profile and stability, antioxidant activities and sensory evaluation of pasta enriched by linseed flour and linseed oil.

Pasta assortments fortified with high quality foods are a modern nutritional trends. This study, explored the effects of fortification with linseed flour (LF) and linseed oil (LO) on durum wheat pasta characteristics. Wheat flour semolina was replaced with 5%, 10% and 15% of LF or 1%, 2.5% and 5% of LO. Control pasta CP (without LF or LO addition), LF-enriched pasta LFP 5%, LFP 10% and LFP 15% and LO-enriched pasta LOP 1%, LOP 2.5% and LOP 5% was compared for the proteins, fat and phenolic contents and fatty acids (FA) profile. Impact on lipid oxidation and sensory evaluation were also determined. Fortification of pasta with LF improved significantly (p < 0.05) the contents of protein, fat and phenolic compared to CP whereas the enrichment of pasta with LO resulted in a significant increase (p < 0.05) in the content of fat and a significant decrease in protein and phenolic contents. All the formulations decreased the saturated FA percent and increased the polyunsaturated FA percent with enhancement of omega-3 FA content. Antioxidant activity measured by FRAP and DPPH assays was improved after the fortification. For lipid oxidation, the replacement of semolina by LF or LO promoted an increase (p < 0.05) on TBARS values in level-dependent manner. Regarding sensory evaluation, the two types of fortification did not affect the taste; flavor and aroma of cooked pasta, but LOP 5% showed the highest score of the overall acceptability. The results recommended the possibility of producing pasta supplemented with LF or LO (even at a level of 15% and 5% respectively) as a functional food.

Effect of acute meal and long-term intake of a Mediterranean Diet providing different amounts of carbohydrates on physical performance and biomarkers in non-professional strength athletes.

The study aims to evaluate the effect of an acute meal and long-term intake of Mediterranean Diet (MD) on different parameters such as strength, physical performance, body composition and blood markers in a group of non-professional athletes who practice a strength activity. Thirteen volunteers completed two 8-week dietary interventions in a randomised, cross-over design. Also an acute study was performed. Subjects received a MD High in carbohydrates, characterised by at least five portions of pasta/week and an average 55-60% of daily energy derived from carbohydrates, versus an MD reduced in carbohydrates, with less than two portions of pasta/week and an average of 40-45% of daily energy provided by carbohydrates. Mainly, data did not show significant differences for the parameters analysed, except for Elbow Flexor maximum voluntary contraction (p = .039). Results enlighten that increasing total carbohydrates intake, as typically in the MD, does not negatively affect physical performance, body composition and strength.

Effect of fibre fortification of low FODMAP pasta.

Irritable bowel syndrome (IBS) is a condition affecting the digestive system and can be triggered by several different factors, including diet. To ease symptoms of IBS, a diet low in fermentable oligo-, di-, monosaccharides and polyols (FODMAPs) is often recommended. Pasta, as a staple food in the Western World, is naturally high in FODMAPs. This study investigates the impact of insoluble and soluble dietary fibre ingredients in low-FODMAPs pasta. The assessment included physicochemical, sensory, and nutritional quality. Soluble fibre strengthened gluten network, which caused a lower cooking loss and a lower release of sugars during in vitro starch digestion. Insoluble fibre interfered with the gluten network development to a higher extent causing a higher sugar release during digestion. This study reveals the most suitable fibre ingredients for the development of pasta with elevated nutritional value and sensory characteristics compared to commercial products on the market. This type of pasta has a high potential of being suitable for IBS patients.

Quality of High-Fibre Pasta Supplemented with Watermelon Rind Powder with Different Particle Sizes.

Research background: Watermelon rind, a by-product of watermelon juice processing, contains large amounts of dietary fibre and phenols with antioxidant capacity. The use of agro-industrial by-products would both improve economic benefits and reduce environmental emissions. The aim of this research is to examine the effect of the particle size of watermelon rind powder on the quality of high-fibre pasta. Experiment approach: The nutritional, physical and physicochemical quality of three samples of watermelon rind powder, sieved through three sieves with aperture size of 400, 210 and 149 µm, were analysed. Durum wheat semolina with watermelon rind powder mass fraction of 10 % were mixed and used to make pasta. Nutritional, textural and cooking quality, sensory acceptability, in vitro glycaemic index and antioxidant bioaccessibility of high-fibre pasta with added watermelon rind powder of different particle sizes were evaluated and compared. Results and conclusions: When the sieve aperture size was reduced from 400 to 149 µm, the soluble dietary fibre and total phenolic contents of watermelon rind powder were increased by 35 and 15 %, respectively, while its insoluble dietary fibre content was decreased by 21 %. Decrease in sieve aperture size from 410 to 149 µm reduced phenolic bioaccessibility of the fortified pasta from 63 to 57 %, but enhanced its predicted glycaemic index from 50 to 69. It also decreased the pasta hardness by 13 %, but improved its elongation rate and tensile strength by 13 and 40 %, respectively. The finer the particles of the watermelon rind powder, the longer the optimal cooking time, the higher the water absorption index, and the lower the cooking loss of the supplemented pasta. Consumers did not notice any significant differences in the overall acceptability among all pasta samples. Novelty and scientific contribution: The particle size of the watermelon rind powder had a major effect on nutritional value, texture and cooking quality of the fortified pasta. In particular, the predicted glycaemic index and antioxidant bioaccessibility of high-fibre pasta were significantly affected by the particle size of the dietary fibre material used in the recipe.

Antinutrient removal in yellow lentils by malting.

BACKGROUND: To improve the current low per capita consumption of lentils, the present study first aimed to minimize the anti-nutrient content of two yellow Moroccan and Italian lentil seeds by resorting to the malting process and then testing the resulting decorticated flours as ingredients in the formulation of gluten-free fresh egg pastas. RESULTS: The most proper operating conditions for the three malting process steps were identified in a bench-top plant. The first (water steeping) and second (germination) steps were studied at 18, 25 or 32 °C. After 2 or 3 h of steeping at 25 °C and almost 24 h of germination, 95-98.8% of the lentil seeds sprouted. By prolonging the germination process to 72 h, the raffinose or phytic acid content was reduced by about 80% or 95% or 27% or 37%, respectively. The third step (kilning) was carried out under fluent dry air at 50 °C for 24 h and at 75 °C for 3 h. The cotyledons of the resulting yellow lentil malts were cyclonically recovered, milled and chemico-physically characterized. CONCLUSION: Both flours were used to prepare fresh egg-pastas essentially devoid of oligosaccharides, and low in phytate (4.6-6.0 mg g-1 ) and in vitro glycemic index (38-41%). However, the cooking quality of the fresh egg pasta made of malted Moroccan lentil flour was higher with respect to its crude protein content and lower with respect to its water solubility index. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

A novel approach to increase calcium and fiber content in pasta using kadamb fruit (Neolamarckia cadamba) powder and study of functional and structural characteristics.

Kadamb is a unique and underutilized fruit having rich nutritional profile. The utilization of kadamb fruit in value addition is very limited. In this study, pasta was made using kadamb fruit powder (KFP). The effect of fortification of KFP on the quality parameters (color, solid loss, percent expansion, hardness, bulk density, and overall acceptability) of pasta was studied. Pasta was prepared using semolina as the base ingredient, and various proportions of KFP (ranging from 0 to 20%) were added for fortification. Dietary fiber and calcium contents of dry pasta were increased from 5.21 ± 0.02 to 15.36 ± 0.02 and 17.57 ± 0.15 to 37.97 ± 0.03, respectively. As the proportion of KFP increased, the cooking time, hardness, and percent solid loss of the cooked pasta also increased. The highest values for overall acceptability, hardness, cooking solid loss, and bulk density were achieved with 10% KFP and 90% semolina were 7.93 ± 0.41, 19.92 ± 0.21 N, 6.30 ± 0.46%, and 331.67 ± 9.60 kg/m3 respectively. Percent expansion of the pasta was noted to be around 98.33 ± 6.5%. The optimal proportion of KFP was found to be 10% for achieving the best overall quality attributes. FTIR (Fourier-transform infrared spectroscopy) and SEM (scanning electron microscopy) analyses were conducted on the pasta, confirming the presence of functional groups and revealing structural changes due to fiber content of KFP. KFP can be used to create functional and nutritious food products, and further research could explore its application in other food formulations as well.

[Study of composite mixtures based on durum wheat semolina and white beans flour for pasta production as specialized food products].

Increasing the number of chronic non-communicable diseases around the world is a critical social problem in many countries. In this regard, the creation of specialized foods that correct dysfunctions of the human body is a priority direction in science and food industry. Legumes are characterized by a high content of protein, minerals and trace elements, which determines their possible use as the main raw materials for creating specialized foods. The aim of this research was to present a medical, biological and technological justification for the possibility of producing pasta based on composite mixtures of durum wheat semolina and white bean flour as specialized foods with increased protein content and optimal mineral ratio. Material and methods. The objects of the study were grains of durum wheat (grade Krasnokutka-13), whole meal flour from white beans, composite mixtures in various percentages, and pasta from them. The nutritional and biological value of durum wheat semolina and bean flour was determined experimentally, the nutrient profile of the developed composite mixtures - by calculation. The rheological properties of dough from composite mixtures and the cooking properties of pasta made from them were assessed. Results. It has been established that increase in the proportion of white bean flour in composite mixes elevated calcium and protein content, optimized an amino acid profile, the ratio of calcium to phosphorus, significantly reduced the glycemic load. Bean flour introduction into composite mixtures positively affected the rheological properties of the dough and products from it, in particular, on the speed and energy intensity of dough kneading and aging processes of starched polysaccharides, however, lead to a decrease in dry pasta strength. The pasta made from composite mixtures meets the technological requirements for pasta in terms of a number of basic indicators, namely: the volume of dry pasta, the volume of pasta after cooking and the boilability coefficient. Conclusion. The rheological and technological properties of composite mixtures and their nutrient profile make it possible to recommend them for the production of pasta, as specialized foods.

Probiotic bacilli incorporation in foods: is really so easy?

Foods containing probiotic bacilli spores are becoming more and more popular because of their resistance to acidic pH, low water activity, and, most importantly, heat processes. Even though spores may engage in some functions, such as immunomodulation, the majority of the beneficial characteristics are unique to vegetative cells. As matter of fact, the development of foods fortified with spores ought to even ensure the germination of the spores along the gastrointestinal tract. In this perspective, vegetative cells derived from spore germination were separately counted on a minimal medium because the traditional approach is based on the use of complex media that allows the spores to germinate independently by stimuli arising from processing or digestion. In more detail, three Bacillus spp. cultures with claimed probiotic properties were added to two entirely unrelated foods (pasta and croissants), and tolerance, as well as germination, was monitored before and after exposure to simulated GIT, as well as at the beginning and end of the products shelf life. For the first time, potential probiotic bacilli were included in a frozen ready-to-bake product. Germination appears to be prevented in this instance, and the impact of baking, matrix and cold storage on spores was examined independently. All of the parameters appeared to contribute, although further research is needed due to the unpredictable behavior exhibited by spores during freezing.

Similar outcomes between transtendon repair and tear completion repair techniques for partial articular-sided supraspinatus tendon avulsion lesions: a systematic review and meta-analysis.

PURPOSE: This study aimed to review studies comparing transtendon repair (TTR) with tear completion repair (TCR) techniques for partial articular-sided supraspinatus tendon avulsion (PASTA) lesions according to postoperative patient-reported outcomes and complications. METHODS: Databases, including PubMed, Embase, Scopus, and Cochrane, were searched for studies published between 2008 and 2022 that directly compared the postoperative patient-reported outcomes and complications of the TTR and TCR techniques for PASTA lesions. Odds ratios (ORs) were calculated for dichotomous outcomes, while mean differences (MDs) were calculated for continuous outcomes. RESULTS: A total of seven studies (497 shoulders) were analysed. No statistically significant differences in the postoperative clinical outcomes at the final follow-up were observed between the TTR and TCR techniques for PASTA lesions. The overall retear rates of the TTR and TCR techniques were 7.7% and 11.6%, respectively (corresponding healing rates were 92.3% and 88.4%), whereas the overall occurrence rates of adhesive capsulitis were 4.7% and 3.3%, respectively. Furthermore, no significant difference was observed in postoperative range of motion (forward flexion, MD = - 1.22, 95% confidence interval (95%CI) - 5.28 to 3.34, n.s.; external rotation, MD = - 1.39, 95% CI - 3.19 to 0.42, n.s.), overall retear rate (OR 0.72, 95% CI 0.29-1.08, n.s.), and occurrence rate of adhesive capsulitis (OR 1.11, 95% CI 0.35-3.52, n.s.) between the two techniques. CONCLUSION: Both techniques improve clinical outcomes while having a low complication rate and a high rate of healing. No significant difference in clinical outcomes was observed between the two techniques. LEVEL OF EVIDENCE: III.

CNN-LSTM Neural Network for Identification of Pre-Cooked Pasta Products in Different Physical States Using Infrared Spectroscopy.

Infrared (IR) spectroscopy is nondestructive, fast, and straightforward. Recently, a growing number of pasta companies have been using IR spectroscopy combined with chemometrics to quickly determine sample parameters. However, fewer models have used deep learning models to classify cooked wheat food products and even fewer have used deep learning models to classify Italian pasta. To solve these problems, an improved CNN-LSTM neural network is proposed to identify pasta in different physical states (frozen vs. thawed) using IR spectroscopy. A one-dimensional convolutional neural network (1D-CNN) and long short-term memory (LSTM) were constructed to extract the local abstraction and sequence position information from the spectra, respectively. The results showed that the accuracy of the CNN-LSTM model reached 100% after using principal component analysis (PCA) on the Italian pasta spectral data in the thawed state and 99.44% after using PCA on the Italian pasta spectral data in the frozen form, verifying that the method has high analytical accuracy and generalization. Therefore, the CNN-LSTM neural network combined with IR spectroscopy helps to identify different pasta products.

Enriched pasta incorporating typical vegetables of mediterranean diet: in vitro evaluation of inhibitory potential on digestive enzymes and predicted glycaemic index.

Swiss chard (Beta vulgaris L.) and chicory (Cichorium intybus L.) contain biologically active compounds with proven health benefits. Durum wheat noodle-shaped pasta enriched with dried and powder leaves of chard or chicory, at two different levels of supplementation (3%, 6%) was prepared on a laboratory scale. The content of polyphenols, pigments, carotenoids, in vitro inhibition of digestive enzymes and the predicted glycaemic response of the fortified pasta were evaluated. All formulations showed in vitro enzyme inhibition of amylase, glucosidase, and lipase and a low pGI <43. The lowest predicted glycaemic index (pGI = 34 ± 1.1) was found for pasta enriched with 3% beet powder. The incorporation of Beta vulgaris and Cichorium intybus leaf powders improved the nutritional properties of the pasta and also imparted an attractive natural colour to the products.

Long-term changes in oral feeding behaviors of growing rats.

Oral feeding is critical for survival in both humans and animals. However, few studies have reported quantitative behavioral measures associated with the development of oral feeding behaviors. Therefore, the present study investigated developmental changes in the oral feeding behaviors of rats by quantitatively assessing pasta eating and licking behaviors. In the pasta eating test, the time to finish pasta sticks of three different thicknesses (Φ = 0.9, 1.4, and 1.9 mm, 4 cm long) was recorded between postnatal day 29 (P29) and P49, because all rats were able to finish eating these pasta sticks on P29. A developmental decrease in the time to finish pasta sticks of all thicknesses was observed during the initial period of recordings and plateaued before P35. The extent of this decrease was dependent on the thickness of pasta sticks. In the licking test, the number of licks per 10 s and the total intake volume during the test were recorded between P19 and P49, because all rats were able to access and lick the solution on P19. The time courses of developmental increases in the number of licks and the total intake volume were similar to the results obtained in the pasta eating test. Collectively, these results suggest that developmental changes in pasta eating and licking behaviors markedly differed between the weanling and periadolescent periods. The present study also demonstrated the applicability of the pasta eating and licking tests to the quantification of developmental changes in the oral feeding behaviors of rats.

Innovative Approaches for Food: Using Natural Phenolic-Rich Extracts to Produce Value-Added Fresh Pasta.

Cereal-based products, which are rich in carbohydrates, are widely consumed worldwide; however, this type of food lacks other nutrients. Phenolic compounds from natural sources, such as Moringa oleifera, can be incorporated into these products to increase their nutritional and biological value. In this study, a phenolic-rich extract was obtained from M. oleifera leaf powder using a Soxhlet extractor. The extract obtained presented a total phenolic content of 79.0 mg of gallic acid equivalents/g and the ABTS and DPPH assays showed that the extract can act as an anti-oxidant agent, with IC50 values of 205.2 mg/L and 636.0 mg/L, respectively. Afterwards, fresh pasta was produced and the extract was incorporated into the pasta to improve its biological properties and extend its shelf-life. The results demonstrated that the addition of M. oleifera to the fresh pasta increased its anti-oxidant capacity and did not interfere with the cooking properties of the product. Moreover, the fortified pasta presented an increased shelf-life, since the extract conferred protection against microbial contamination for longer periods of time. Therefore, these findings showed that the incorporation of phenolic-rich extracts from natural sources (such as M. oleifera) is a feasible sustainable biotechnological approach to produce value-added cereal-based products.

Common Wheat Pasta Enriched with Ultrafine Ground Oat Husk: Physicochemical and Sensory Properties.

Oat husk (hull) is a byproduct of oat processing that is rich in insoluble fiber. The aim of the study was to evaluate the effect of partially replacing wheat flour with oat husk (at levels of 0, 5, 10, 15, and 20 g/100 g) on the physicochemical properties and sensory acceptance of pasta. Additionally, UPLC-MS/MS analysis was performed to identify phenolic acids and flavonoid compounds, and the cooking properties of the pasta were evaluated. The test results indicate that oat husk significantly (p < 0.05) increased the ash and fiber contents in the pasta, while decreasing the protein and fat contents. Moreover, the redness and yellowness of both raw and cooked pasta increased, while lightness decreased as a result of pasta enrichment with oat husk. Oat husk also led to a decrease in the stretching force of cooked samples, although cooking loss increased significantly but did not exceed 8%. The contents of phenolics and antioxidant activity significantly increased with the incorporation of hull in pasta recipes. UPLC-MS/MS analysis showed that the enriched pasta was especially abundant in ferulic acid. Products with up to 10 g of husk/100 g of wheat flour showed good consumer acceptance. However, higher levels of this additive led to notably lower assessments, particularly in terms of pasta texture.

Varietal identification in pasta through an SSR-based approach: a case study.

BACKGROUND: Pasta is a worldwide popular Italian food made exclusively of durum wheat. The choice of variety to be used to produce pasta is at the discretion of the producer based on the peculiar characteristics of each cultivar. The availability of analytical approaches for the tracking of specific varieties along the productive chain is becoming increasingly important to authenticate the pasta products and distinguish between fraudulent activities and cross-contaminations during the production process. Among the different methods, molecular approaches based on DNA markers are the most used for these purposes because of their ease of use and high reproducibility. RESULTS: In the present study, we used an easy simple sequence repeats-based method to identify the durum wheat varieties used to produce 25 samples of semolina and commercial pasta comparing their molecular profile with those of the four varieties declared by the producer and other 10 durum wheat cultivars commonly used in pasta production. All of the samples showed the expected molecular profile; however, most of them present also a foreign allele indicating a possible cross-contamination. Moreover, we evaluated the accuracy of the proposed approach through the analysis of 27 hand-made mixtures with increasing amounts of a specific contaminant variety, allowing the estimation of the limit of detection of 5% (w/w). CONCLUSION: We demonstrated the feasibility of the proposed method and its effectiveness in the detection of not declared varieties when these are present in a percentage equal to or higher than 5%. © 2023 The Authors. Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Nutritional, textural and sensory properties of lasagna dough containing air yam, a non-conventional edible plant.

The incorporation of non-conventional edible plants into gluten-free food products shows potential and the ability to increase nutritional properties. Therefore, the aim of this study was to develop a gluten-free air yam-based lasagna dough and to evaluate the nutritional, textural, and sensory properties. First, the air yam flour was obtained by drying the tubers at 55 °C for 16 h and then, mixed with the water and egg powder, it constituted the lasagna dough. The nutritional composition and bioaccessible mineral content of air yam flour and lasagna dough were evaluated, as well as the texture, microbiological and sensory properties of the lasagna dough. The results indicated that air yam flour and the lasagna dough had high dietary fiber contents, 17.1% and 9.4%, respectively. Additionally, the low-fat content found means that the lasagna dough can be considered both gluten-free and fat-free. The most bioaccessible mineral present was K in both air yam flour and lasagna dough. In sensory analysis, the lasagna dough containing air yam showed an intermediate acceptability in relation to two already commercialized lasagna doughs. This study shows an application to this group of plants that are still little explored, allowing the development to consumers and industries.

GpsB Promotes PASTA Kinase Signaling and Cephalosporin Resistance in Enterococcus faecalis.

Enterococci are opportunistic pathogens that can cause severe bacterial infections. Treatment of these infections is challenging because enterococci possess intrinsic and acquired mechanisms of resistance to commonly used antibiotics, including cephalosporins. The transmembrane serine/threonine PASTA kinase, IreK, is an important determinant of enterococcal cephalosporin resistance. Upon exposure to cephalosporins, IreK becomes autophosphorylated, which stimulates its kinase activity to phosphorylate downstream substrates and drive cephalosporin resistance. However, the molecular mechanisms that modulate IreK autophosphorylation in response to cell wall stress, such as that induced by cephalosporins, remain unknown. A cytoplasmic protein, GpsB, promotes signaling by PASTA kinase homologs in other bacterial species, but the function of enterococcal GpsB has not been previously investigated. We used in vitro and in vivo approaches to test the hypothesis that enterococcal GpsB promotes IreK signaling in response to cephalosporins to drive cephalosporin resistance. We found that GpsB promotes IreK activity both in vivo and in vitro. This effect is required for cephalosporins to trigger IreK autophosphorylation and activation of an IreK-dependent signaling pathway, and thereby is also required for enterococcal intrinsic cephalosporin resistance. Moreover, analyses of GpsB mutants and a ΔireK gpsB double mutant suggest that GpsB has an additional function, beyond regulation of IreK activity, which is required for optimal growth and full cephalosporin resistance. Collectively, our data provide new insights into the mechanism of signal transduction by the PASTA kinase IreK and the mechanism of enterococcal intrinsic cephalosporin resistance. IMPORTANCE Enterococci are opportunistic pathogens that can cause severe bacterial infections. Treatment of these infections is challenging because enterococci possess intrinsic and acquired resistance to commonly used antibiotics. In particular, enterococci are intrinsically resistant to cephalosporin antibiotics, a trait that requires the activity of a transmembrane serine/threonine kinase, IreK, which belongs to the bacterial PASTA kinase family. The mechanisms by which PASTA kinases are regulated in cells are poorly understood. Here, we report that the cytoplasmic protein GpsB directly promotes IreK signaling in enterococci to drive cephalosporin resistance. Thus, we provide new insights into PASTA kinase regulation and control of enterococcal cephalosporin resistance, and suggest that GpsB could be a promising target for new therapeutics to disable cephalosporin resistance.

Lowering the predicted glycemic index of pasta using dried onions as functional ingredients.

Pasta is a commonly consumed food; adding some ingredients, maybe turn it into a functional food with health benefits. These ingredients consist of dietary fiber, antioxidant molecules, and enzyme inhibitor compounds, related to a reduced risk for some diseases. Onion (Allium cepa L.) is a rich source of bioactive compounds rendering it a relevant candidate for the production of functional foods. The present study examines the in vitro starch digestibility of durum wheat pasta supplemented with 3% onion flour. The incorporation of onion flour attenuated the extent of starch digestion and accordingly the area under the curve of reducing sugars discharged during in vitro digestion. The predicted glycemic index (pGI) of pasta supplemented with onion flour (3%) was significantly lower (pGI = 54 ± 0.17) than the control pasta (pGI = 72 ± 0.14). These results indicate that 3% onion fortified pasta represents a food with potential healthy properties, showing glucose-lowering capabilities in vitro.