Quality Evaluation of Fresh Pasta Fortified with Sourdough Containing Wheat Germ and Wholemeal Semolina.

Pasta is a staple food in the Mediterranean diet, primarily manufactured with two essential ingredients, semolina and water; nowadays, it is often supplemented with functional ingredients. In this work, a sourdough obtained with wheat germ and wholemeal semolina was used, in order to improve sensorial and nutritional properties of fresh pasta, to prevent lipids oxidation, and to improve the shelf life. Three different formulations were prepared, a first one using semolina, a second one with raw wheat germ, wholemeal semolina, and semolina, and the last one with semolina and sourdough. The study highlighted the improved nutritional properties of pasta with sourdough (reduced phytic acid content, higher antioxidant activity and phenolic content). Proteins, ashes, dietary fibers, lipids, and tocols (vitamin E) increased in pasta with wheat germ and wholemeal semolina, and with sourdough. The amount of tocols decreased in pasta samples after cooking, except for the ß-tocopherol in sourdough pasta, the amount of which remained high, surprisingly. Lipase and lipoxygenase enzymes likely decreased as an effect of the pasteurization process. The NMR analysis showed that lipid oxidation was higher in semolina pasta than in pasta with wheat germ, most likely due to the protective effect of antioxidants deriving from wheat germ.

From Hop to Beer: Influence of Different Organic Foliar Fertilisation Treatments on Hop Oil Profile and Derived Beers' Flavour.

Foliar fertilisation is known to influence the physiological response of Humulus lupulus (hop plants), but its effect on the flavour profile of beer still has to be investigated. By comparing the effects of four fertilisation treatments, this study aims at determining whether different foliar fertilisation treatments have a significant impact on hop plants' aromatic quality and that of the beer produced. Hop cones harvested from each experimental treatment were brewed to obtain five single dry-hopped beers, which were subsequently analysed. Gas chromatography-mass spectrometry (GC-MS) and electronic nose (Cyranose 320) analyses were performed on the hop cones, while headspace solid-phase microextraction-gas chromatography-mass spectrometry HS-SPME-GC-MS, electronic nose and sensory analyses were carried out on the beers produced. The analyses not only allowed for a differentiation between the hops from the four fertilisation treatments and the control but also enabled a differentiation between the beers produced for their identification. Sensory evaluation revealed consumer preferences regarding the dry-hopped beers analysed, evidencing their distinctive features, including significant differences in both aroma and flavour.

In Vivo and In Vitro Starch Digestibility of Fresh Pasta Produced Using Semolina-Based or Wholemeal Semolina-Based Liquid Sourdough.

The use of wholemeal flour and sourdough fermentation in different food matrices has received considerable attention in recent years due to its resulting health benefits. In this study, a semolina-based and a wholemeal semolina-based sourdough were prepared and added to the formulation of gnocchetti-type fresh pasta. Four types of gnocchetti were made, using semolina plus semolina-based sourdough (SS), semolina plus wholemeal semolina-based sourdough (SWS), semolina alone (S), and semolina plus wholemeal semolina (WS). The latter two were used as controls. The digestibility of starch was studied both in vitro and in vivo, and the glycemic response (GR) and glycemic load (GL) were determined. Starch digestibility, both in vivo and in vitro, was higher in wholemeal semolina than semolina pasta and the resulting GR values (mg dL-1 min-1) were also higher (2209 and 2277 for WS and SWS; 1584 and 1553 for S and SS, respectively). The use of sourdough significantly reduced the rapidly digestible starch (RDS) content and increased the inaccessible digestible starch (IDS) content. The addition of sourdough to the formulation had no effect on the GR values, but led to a reduction of the GL of the pasta. These are the first data on the GR and GL of fresh pasta made with sourdough.

Impact of Almond Variety on "Amaretti" Cookies as Assessed through Image Features Modeling, Physical Chemical Measures and Sensory Analyses.

In the present study, the influence of almond variety on color, chemical, physical and sensory characteristics of "amaretti" cookies during the shelf life, was assessed. Four varieties were chosen for the study, two of which were local (Cossu, Arrubia) and two widely cultivated (Tuono, Texas). Almonds have been characterized in the content of proteins, crude fat, amygdalin and fatty acids profile. The evolution of the characteristics during the shelf life hasbeen measured through image data modeling, texture, physical chemical and sensory analyses. Data were then treated with a multivariate approach performing a PCA. Image analysis and fitting on log normal and powerlaw functions highlighted the influence of the variety on the total area affected by surface breakages, and on the distribution of the cracking surfaces dimension classes. Texture parameters (crust hardness, thickness and work of deformation) were negatively correlated to moisture content. Sensory profile confirmed the differences in tactile features measured through instrumental texture, while slight to no differences were found in odor profile. Consumer test showed an higher acceptability for Arrubia, Texas and Tuono samples throughout the shelf life, while Cossu samples were less accepted. Overall, the choice of almond variety influences product features and liking of almond products, therefore it represents an important phase to direct the choice of both farmers and confectionery manufacturers.

Fresh Pasta Manufactured with Fermented Whole Wheat Semolina: Physicochemical, Sensorial, and Nutritional Properties.

Fresh pasta (SP) was prepared by mixing semolina with liquid sourdough, whole wheat semolina based, and the effects of sourdough inclusion were evaluated against a control sample (CP) prepared using semolina and whole wheat semolina. Physicochemical, nutritional, and sensorial analyses were performed on pasteurized fresh pasta, before and after cooking. The optimum cooking time was not affected by whole wheat sourdough, whereas differences were found in color, firmness, and cooking loss. Changes of in vitro digested starch fractions in SP pasta were affected by a higher cooking loss. Overall, SP samples were characterized by improved nutraceutical features, namely higher content of free essential amino acids and phenolic compounds, lower phytic acid content, and higher antioxidant activity. Sensory analyses (acceptability and check-all-that-apply (CATA) tests) showed significantly higher scores for the SP, and the differences were enhanced when the consumers were informed about the product composition and how it was manufactured. Consumers checked for more positive sensory parameters for the SP than the CP.

Effect of liquid sourdough technology on the pre-biotic, texture, and sensory properties of a crispy flatbread.

The present study compared liquid sourdough technology with baker's yeast leavening when applied to the production of a semolina-based crispy flatbread. Following in vitro starch digestion, the results revealed the sourdough leavened flatbread to contain a lower percentage of rapidly digestible starch (16%), higher amounts of slowly digestible starch (27%) and inaccessible digestible starch (4.1%) compared with the baker's yeast leavened flatbread (20, 20, and 2.4%, respectively), making the former nutritionally healthier. The sourdough leavened bread was crispier, stiffer and more solid, as shown by texture analyses, although Raman spectroscopy revealed no differences in the crystallinity status of starch. The descriptive analyses show that the use of sourdough enhances the positive sensory traits, as rated by the consumer panel scores (6.08 vs. 5.56). In summary, the results indicate that the implementation of sourdough technology in the production of flat crispy breads could confer economic advantages to this product.

Acanthamoeba castellanii Genotype T4 Stimulates the Production of Interleukin-10 as Well as Proinflammatory Cytokines in THP-1 Cells, Human Peripheral Blood Mononuclear Cells, and Human Monocyte-Derived Macrophages.

Free-living amoebae of the genus Acanthamoeba can cause severe and chronic infections in humans, mainly localized in immune privileged sites, such as the brain and the eye. Monocytes/macrophages are thought to be involved in Acanthamoeba infections, but little is known about how these facultative parasites influence their functions. The aim of this work was to investigate the effects of Acanthamoeba on human monocytes/macrophages during the early phase of infection. Here, THP-1 cells, primary human monocytes isolated from peripheral blood, and human monocyte-derived macrophages were either coincubated with trophozoites of a clinical isolate of Acanthamoeba (genotype T4) or stimulated with amoeba-derived cell-free conditioned medium. Production of proinflammatory cytokines (tumor necrosis factor alpha [TNF-α], interleukin-6 [IL-6], and IL-12), anti-inflammatory cytokine (IL-10), and chemokine (IL-8) was evaluated at specific hours poststimulation (ranging from 1.5 h to 23 h). We showed that both Acanthamoeba trophozoites and soluble amoebic products induce an early anti-inflammatory monocyte-macrophage phenotype, characterized by significant production of IL-10; furthermore, challenge with either trophozoites or their soluble metabolites stimulate both proinflammatory cytokines and chemokine production, suggesting that this protozoan infection results from the early induction of coexisting, opposed immune responses. Results reported in this paper confirm that the production of proinflammatory cytokines and chemokines by monocytes and macrophages can play a role in the development of the inflammatory response during Acanthamoeba infections. Furthermore, we demonstrate for the first time that Acanthamoeba stimulates IL-10 production in human innate immune cells, which might both promote the immune evasion of Acanthamoeba and limit the induced inflammatory response.

Anticyclic citrullinated protein antibodies are implicated in the development of cardiovascular disease in rheumatoid arthritis.

OBJECTIVE: Previous studies have suggested a relationship between anticyclic citrullinated protein (CCP) levels and development of cardiovascular disease in rheumatoid arthritis (RA). However, a limited number of studies have demonstrated an involvement of anti-CCPs in those processes. This study was aimed to define the specific role of these auto-antibodies in the pro-oxidative, inflammatory, and proatherogenic profile observed in leukocytes from RA patients. APPROACH AND RESULTS: Seventy-five RA patients and 31 healthy donors were enrolled. Carotid intima media thickness was evaluated as atherosclerosis marker. Several procoagulant and inflammatory factors, leukocyte activation, and oxidative stress markers were analyzed in plasma and leukocyte subsets. Anti-CCPs were purified from plasma of RA patients, and in vitro treatment of healthy leukocytes was conducted. High titers of anti-CCPs were associated to altered expression of prothrombotic and inflammatory markers, high oxidative stress, and pathological carotid intima media thickness in RA patients. Notably, gene expression analysis showed that lymphocytes were major players in altered inflammatory profile, monocytes were responsible for the protrombotic and atherogenic status, and neutrophils mainly displayed a pro-oxidative feature. In vitro treatment with purified anti-CCPs fully recapitulated that pathogenic profile, promoting the activation of leukocytes. CONCLUSIONS: Anti-CCPs are key players in the inflammatory and proatherogenic status of RA patients. The effects are specific of the immune cell targeted, promoting overexpression of thrombotic, inflammatory, and pro-oxidative markers in monocytes; pro-oxidative status in neutrophils; and proinflammatory profile in lymphocytes. Targeting these autoantibodies would be an excellent strategy to prevent the development of cardiovascular disease in RA.

Regular cigarette smoking influences the transsulfuration pathway, endothelial function, and inflammation biomarkers in a sex-gender specific manner in healthy young humans.

Cigarette smoking (CS) is the primary cause of preventable morbidity and mortality. Abundant clinical evidence suggests that CS is more harmful to women; however, the mechanisms responsible for these differences are not yet known. CS alters endothelial function, the redox state, inflammation, and global DNA methylation, which is associated with one-carbon metabolism and the transsulfuration pathway. However, it is not known whether the previously identified alterations are sex-gender related. Healthy adult men and oral contraceptive-free women with regular menstrual cycles were enrolled; women were examined during the follicular phase. Men had higher plasma levels of uric acid, total bilirubin, homocysteine, glutamylcysteine, total glutathione, cysteinylglycine; had more monocytes and released more TNF-alpha from human monocytes derived macrophages (hMDMs), but they had fewer platelets and lower levels of DNA methylation, and their hMDMs released less TNF-alpha after LPS stimulation. MDA, taurine, cysteine, arginine, ADMA, and SDMA were not different. CS decreased global DNA methylation more in women and increased the platelet, monocyte, and lymphocyte counts and the homocysteine, arginine, and ADMA levels only in women, whereas increased the neutrophil and eosinophil counts only in men. Additionally, CS reduced the sex-gender differences in total bilirubin, basal and LPS-induced TNF-alpha release, total glutathione, and glutamylcysteine, leaving unchanged cysteinylglycine, taurine, SDMA, MDA, and cysteine. These data suggest that cardiovascular risk factors seem to come earlier in young healthy female smokers than in young healthy male smokers, supporting the greater alarmism regarding the effects of CS in women and providing a basis for understanding the sex-gender differences. These results also suggest that cessation programs targeting women are needed.

Oral contraceptives modify DNA methylation and monocyte-derived macrophage function.

BACKGROUND: Fertile women may be encouraged to use contraception during clinical trials to avoid potential drug effects on fetuses. However, hormonal contraception interferes with pharmacokinetics and pharmacodynamics and modifies internal milieus. Macrophages depend on the milieu to which they are exposed. Therefore, we assessed whether macrophage function would be affected by the use of combined oral contraceptives (OCs) and if this influence depended on the androgenic or non-androgenic properties of progestin. METHODS: Healthy adult women were enrolled and stratified into two groups: women who did not use OCs (Fs) and women treated with OCs (FOCs). FOCs were further stratified as a function of androgenic (FOCA+) and non-androgenic (FOCA-) properties of progestins. Routine hematological, biochemical, inflammatory and endothelial dysfunction parameters were measured. Monocyte-derived macrophages (MDMs) were evaluated for the expression and activity of estrogen receptors and androgen receptors, and release of tumor necrosis factor α (TNFα) was measured from unstimulated and lipopolysaccharide-stimulated cells. RESULTS: As is already known, the use of OCs changed numerous parameters: the number of lymphocytes, iron levels, total iron-binding capacity of transferrin, triglycerides, high-density lipoprotein, total cholesterol, and C-reactive protein increased, while prothrombin time and alkaline phosphatase decreased. Hormonal levels also varied: cortisol was higher in FOCs, while luteinizing hormone, follicle-stimulating hormone, and testosterone were lower in FOCs. Asymmetric dimethylarginine, an index of endothelial function, was lower in FOC than in Fs, as were cysteine and bilirubin. The androgenic properties of progestins affected the activity of OCs: in particular, white blood cell count, hemoglobin, high-density lipoprotein and calcium were higher in FOCA- than in FOCA+, whereas percentage oxygen saturation and γ-glutamyl transpeptidase were lower in FOCA- than in FOCA+. Importantly, FOCs had a lower global DNA methylation, indicating that OC may have epigenetic effects on gene expression. OC did not modify the expression of androgen receptor but increased estrogen receptor α expression, more considerably in FOCA+, and decreased estrogen receptor ß, more considerably in FOCA-. Importantly, the activation state of estrogen receptor ß in FOCs was decreased, while estrogen receptor α was not active in either Fs or FOCs. Unstimulated MDMs obtained from FOCs showed higher release of TNFα in comparison with Fs. After lipopolysaccharide stimulation, the release of TNFα was significantly higher in Fs than in FOCs. CONCLUSIONS: OC use induced many changes in hematological and plasmatic markers, modifying hormonal levels, endothelial function, inflammation index and some redox state parameters, producing a perturbation of the internal milieu that impacted macrophagic function. In fact, different levels of estrogen receptor expression and release of TNFα were observed in macrophages derived from OC users. Some of the above activities were linked to the androgenic properties of progestin. Even though it is not known whether these effects are reversible, the results indicate that to avoid potential skewing of results only a single type of OC should be used during a single clinical trial.

Quantification of histidine, 1-methylhistidine and 3-methylhistidine in plasma and urine by capillary electrophoresis UV-detection.

We describe a new CE method with UV-detection for the quantification of histidine (His) and its methylated forms 1-methylhistidine and 3-methylhistidine, both in plasma and urine. Analytes were basically resolved using a 60 mmol/L Tris-phosphate run buffer pH 2.2 in less than 12 min. The use of a mixture of ACN/ammonia (80:20) for protein precipitation allows the quantitative recovery of all His from plasma. The optimization of the sample volume injection permits to reach an LOD of 20 nmol/L, thus improving the sensitivity of about hundred times in comparison to the previous described assays. Moreover, the opportunity to also measure creatinine in the same run makes it possible to evaluate the renal function contemporarily, thus avoiding further dosages with significant time saving. The application method has been proved by measuring His, 1-methylhistidine and 3-methylhistidine in 44 healthy subjects. In conclusion, our new method seems to be an inexpensive, fast and specific tool to assess large numbers of patients for routine analysis both in clinical and research laboratories.

Quantification of neurotransmitter amino acids by capillary electrophoresis laser-induced fluorescence detection in biological fluids.

The role of neurotransmitter amino acids (NAAs) in the functioning of the nervous system has been the focus of increasingly intense research over the past several years. Among the various amino acids that have important roles as neurotransmitters, there are alanine (Ala), glutamic acid (Glu), aspartic acid (Asp), serine (Ser), taurine (Tau) and glycine (Gly). NAAs are present in plasma, cells and--at trace levels--in all biological fluids, but complex components in biological matrices make it difficult to determine them in biological samples. We describe a new capillary electrophoresis (CE) method with laser-induced fluorescence detection by which analytes are resolved in less than 12 minutes in a 18 mmol/L phosphate run buffer at pH 11.6. The use of elevated temperatures during sample derivatization leads to a drastic reduction in the reaction time, down to 20 min, compared to the 6-14 h usually described for reactions between FITC and amino acids at room temperature. In order to demonstrate its wide range of applications, the method was applied to the analysis of NAA in human plasma and in other sample types, such as red blood cells, urine, cultured cells, cerebrospinal fluid, saliva and vitreous humor, thus avoiding the typical limitations of other methods, which are normally suitable for use with only one or two matrix types.

Determination of homocysteine thiolactone, reduced homocysteine, homocystine, homocysteine-cysteine mixed disulfide, cysteine and cystine in a reaction mixture by overimposed pressure/voltage capillary electrophoresis.

An elevated level of thiol amino acid homocysteine is associated with several complex disorders. Homocysteine ability to bind proteins, thereby modulating their structure and function, is proposed to be one of the mechanisms of homocysteine induced pathogenecity. Homocysteine and homocysteine thiolactone bind to protein cysteine and lysine residues respectively. A major hurdle in studying protein homocysteinylation is the lack of suitable analytical techniques to determine simultaneously the concentrations of reduced and oxidized forms of homocysteine and cysteine (especially homocysteine-cysteine mixed disulfide) together with thiolactone formed during the reaction of homocysteine or thiolactone with proteins. Herein we report a capillary electrophoresis method to determine simultaneously the levels of these intermediates. For this 40 mmol/L Tris phosphate buffer at (pH 1.60) was used as running electrolyte, and the separation was performed by the simultaneous application of a CE voltage of 15kV and an overimposed pressure of 0.1 psi. A rapid separation of these intermediates in less than 6 min with a good reproducibility of both peak areas (CV<2%) and migration time (CV<0.2%) was obtained. The applicability of our method was validated by incubating reduced homocysteine and albumin and measuring the reaction intermediates in the solution mixture.

Increased low-density lipoprotein S-homocysteinylation in chronic kidney disease.

BACKGROUND: Since low-density lipoprotein (LDL) S-homocysteinylation has been recently reported to enhance atherogenicity of lipoprotein, we have investigated the levels of homocysteine (Hcy) linked to LDL in chronic proteinuric patients in which lipid abnormalities highly contribute to the excess of morbidity and mortality. METHODS: We used capillary electrophoresis to measure LDL-bound thiol Hcy, cysteine (Cys), cysteinylglycine (Cys-Gly), glutathione (GSH), and glutamylcysteine (Glu-Cys) in 30 chronic kidney disease (CKD) individuals and 60 healthy volunteers. RESULTS: We found more elevated levels of total plasma Hcy, Cys, GSH and Glu-Cys in patients than in controls and also found that Hcy and Cys bound to LDL were significantly increased in nephropathic subjects. By multiple linear regression, we found that in healthy people, total Hcy was the most important determinant of LDL-bound Hcy and Cys-Gly was negatively associated with apoB-Hcy concentrations. In CKD the most important determinant of homocysteinylation was creatinine while total plasma Hcy is weakly associated with apoB-Hcy. CONCLUSIONS: The increased levels in Hcy-LDL observed in CKD patients might account, at least in part, for the excess of cardiovascular risk; thus LDL S-homocysteinylation can be considered a key marker of risk for cardiovascular disease in these individuals.

Ultra-fast adenosine 5'-triphosphate, adenosine 5'-diphosphate and adenosine 5'-monophosphate detection by pressure-assisted capillary electrophoresis UV detection.

Herein, we report a new CE method to measure adenine nucleotides adenosine 5'-triphosphate, adenosine 5'-diphosphate, and adenosine 5'-monophosphate in red blood cells. For this purpose, 20 mmol/L sodium acetate buffer at pH 3.80 was used as running electrolyte, and the separation was performed by the simultaneous application of a CE voltage of 25 kV and an overimposed pressure of 0.2 psi from inlet to outlet. A rapid separation of these analytes in less than 1.5 min was obtained with a good reproducibility for intra- and inter-assay (CV<4 and 8%, respectively) and an excellent analytical recovery (from 98.3 to 99%). The applicability of our method was proved by measuring adenine nucleotides in red blood cells.

Improved rapid assay of plasma uric acid by short-end injection capillary zone electrophoresis.

A rapid and simple short-end injection capillary zone electrophoresis method was developed for the quantification of plasma uric acid. The separation was performed in an uncoated fused-silica capillary (50 microm ID, 60 cm total length, 10.2 cm effective length) by using as a background electrolyte a 75 mmol/L glycylglycine solution titrated with NaOH 5 mol/L to pH 9.0, a voltage of 28 kV, a cartridge temperature of 15 degrees C, and direct UV detection at 292 nm. Under optimized conditions, uric acid was determinate in little more than 1 min (1.076 minutes). In order to verify the accuracy of the analysis, urate levels were measured in 543 apparently healthy volunteers by the new assay and our previous method, and the obtained data were compared by Passing-Bablock regression, Bland-Altman test, and a new regression-based approach, which showed a good agreement between two methods.

Sourdough-leavened bread improves postprandial glucose and insulin plasma levels in subjects with impaired glucose tolerance.

Sourdough bread has been reported to improve glucose metabolism in healthy subjects. In this study postprandial glycaemic and insulinaemic responses were evaluated in subjects with impaired glucose tolerance (IGT) who had a meal containing sourdough bread leavened with lactobacilli, in comparison to a reference meal containing bread leavened with baker yeast. Sixteen IGT subjects (age range 52-75, average BMI 29.9 +/- 4.2 kg/ m2) were randomly given a meal containing sourdough bread (A) and a meal containing the reference bread (B) in two separate occasions at the beginning of the study and after 7 days. Sourdough bread was leavened for 8 h using a starter containing autochthonous Saccharomyces cerevisiae and several bacilli able to produce a significant amount of D-and L-lactic acid, whereas the reference bread was leavened for 2 h with commercial baker yeast containing Saccharomyces cerevisiae. Plasma glucose and insulin levels were measured at time 0, 30, 60, 120, and 180 min. In IGT subjects sourdough bread induced a significantly lower plasma glucose response at 30 minutes (p = 0.048) and a smaller incremental area under curve (AUC) delta 0-30 and delta 0-60 min (p = 0.020 and 0.018 respectively) in comparison to the bread leavened with baker yeast. Plasma insulin response to this type of bread showed lower values at 30 min (p = 0.045) and a smaller AUC delta 0-30 min (p = 0.018). This study shows that in subjects with IGT glycaemic and insulinaemic responses after the consumption of sourdough bread are lower than after the bread leavened with baker yeast. This effect is likely due to the lactic acid produced during dough leavening as well as the reduced availability of simple carbohydrates. Thus, sour-dough bread may potentially be of benefit in subjects with impaired glucose metabolism.

Molecular characterization of lactic acid bacteria from sourdough breads produced in Sardinia (Italy) and multivariate statistical analyses of results.

The objective of this work was to investigate the structure and diversity of lactic acid bacteria (LAB) communities in sourdough used for the production of traditional breads (Carasau, Moddizzosu, Spianata, Zichi) in Sardinia. 16S rDNA sequencing and Randomly Amplified Polymorphic DNA (RAPD-PCR) was applied for the identification and typing of the LAB isolated from 25 samples of sourdoughs. Multivariate statistical techniques were applied to RAPD-PCR pattern to study the biological diversity of sourdough samples. Twelve different species of LAB were identified, and most isolates were classified as facultative heterofermentative lactobacilli. Lactobacillus pentosus dominated the lactic microflora of many samples while Lactobacillus sanfranciscensis was isolated only from a limited number of samples. Although heterofermentative species represented between between 30% and 60% of the isolates in Carasau, Spianata and Zichi sourdoughs, only 2% of the isolates from Moddizzosu sourdoughs were identified as heterofermentative LAB. RAPD-PCR with a single primer followed by cluster analysis did not allow the identification of the isolates at the species level. However, a multidimensional scaling/bootstrapping approach on the RAPD-PCR patterns uncovered the diversity of the LAB communities of LAB showing differences both within and between bread types.