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1.
Food Chem ; 401: 134185, 2023 Feb 01.
Article in English | MEDLINE | ID: mdl-36113218

ABSTRACT

Alternative sources of edible proteins are required to feed the world's growing population, such as Moringa oleifera leaves, a protein source with a balanced amino acid composition. Since Moringa leaf proteins is a novel food in the EU and UK, an assessment of their potential allergenicity of is required. Proteins from Moringa leaf powder were characterised using traditional proteomic approaches. The proteins identified were evaluated for their allergenic potential using in-silico tools. The main proteins identified belonged to photosynthetic and metabolic pathways. In-silico analysis of the leaf proteome identified moritides as potential allergens by homology with a latex allergen implicated in fruit-latex syndrome. This analysis also identified a nsLTP, a major panallergen in food. The presence of these putative allergens was confirmed by de-novo sequencing. Our study allowed identification of putative allergens, Morintides and nsLTP. Further in-vitro and in-vivo investigations are required to confirm their allergenic potential.


Subject(s)
Food Ingredients , Moringa oleifera , Allergens/chemistry , Moringa oleifera/chemistry , Proteomics , Proteome/metabolism , Powders/metabolism , Plant Proteins/metabolism , Plant Leaves/genetics , Plant Leaves/metabolism , Amino Acids/metabolism
2.
Food Res Int ; 154: 111012, 2022 04.
Article in English | MEDLINE | ID: mdl-35337570

ABSTRACT

The demand for sustainably produced proteins is increasing with the world population and is prompting a dietary shift toward plant sourced proteins. Vegetable proteins have lower digestibility and biological value compared to animal derived counterparts. We explored sprouting of chickpea seeds as a strategy for improving digestibility. Protein evolution associated with by the sprouting process was assessed by proteomics. The sprouting induced breakdown of seed storage proteins and doubled the release of free alpha-amino nitrogen in sprouted chickpea flour. During sprouting, several enzymes involved in plant development were newly expressed. An ex vivo model of gastroduodenal and jejunal digestion was applied to assess the bioaccessibility of the protein digests. Proteins from chickpea sprouts showed a greater susceptibility to digestion with a 10% increase in alpha amino nitrogen. Peptides with potential immunoreactivity or bioactivity were catalogued in both digested chickpea sprouts and seeds using an in-silico approach. Peptides belonging to the non-specific transfer proteins, which are allergens in pulses, and peptides belonging to an IgE-binding hemagglutinin protein could only be identified in the digested chickpea sprouts. The observation collected paved the way to immune-based evaluations to assess the effect of germination on the allergenic potential.


Subject(s)
Cicer , Animals , Digestion , Flour , Microvilli , Proteome/metabolism
3.
Food Res Int ; 148: 110617, 2021 10.
Article in English | MEDLINE | ID: mdl-34507761

ABSTRACT

Wheat, an essential ingredient for several bakery preparations, is also responsible for gluten-related diseases in sensitive subjects. The effect of the N fertilization rate (80 vs 160 kg N ha-1) on gluten protein expression profile has been evaluated considering two soft wheats (landrace and modern) and one tritordeum cultivar (cv), grown in the same experimental field in North Italy. The proteins of refined flour were characterized through advanced proteomic approaches, including chromatography (RP-HPLC) and electrophoresis. A static model system was used to simulate in vitro digestion and the digestome peptides were examined by mass spectrometry and in silico approaches, to investigate the celiac and allergenic sequences. The CD-toxic epitopes in the digested samples were quantified by means of a R5 ELISA assay. The N fertilization rate increased the grain protein content, but it did not lead to any difference in gluten composition, with exception of glu/glia ratio in the modern wheat cv. Moreover, the gluten composition and the occurrence of toxic/allergenic epitopes varied to a great extent, according mostly to the genotype. A lower immunoreactivity, determined using R5 ELISA, was detected for the digested tritordeum flours than for the landrace (-51%) or modern (-58%) cvs, while no significant difference was observed for the N rates between each genotype. In silico analysis showed that tritordeum has fewer CD epitopes belonging to the ω-gliadins and a lower LMW-GS than the landrace or modern cv. Tritordeum presented fewer α-gliadin allergenic epitopes than the modern wheat cv. The lower frequency of celiac epitopes in tritordeum, compared to the old and the modern wheat, is probably due to the absence of a D genome.


Subject(s)
Celiac Disease , Triticum , Fertilization , Humans , Nitrogen , Proteomics
4.
Clin Genitourin Cancer ; 18(6): e698-e704, 2020 12.
Article in English | MEDLINE | ID: mdl-32493676

ABSTRACT

INTRODUCTION: The objective of this study was to test Prostate Imaging Reporting and Data System (PI-RADS) classification on multiparametric magnetic resonance imaging (mpMRI) and MRI-derived prostate-specific antigen density (PSAD) in predicting the risk of reclassification in men in active surveillance (AS), who underwent confirmatory or per-protocol follow-up biopsy. MATERIALS AND METHODS: Three hundred eighty-nine patients in AS underwent mpMRI before confirmatory or follow-up biopsy. Patients with negative (-) mpMRI underwent systematic random biopsy. Patients with positive (+) mpMRI underwent targeted fusion prostate biopsies + systematic random biopsies. Different PSAD cutoff values were tested (< 0.10, 0.10-0.20, ≥ 0.20). Multivariable analyses assessed the risk of reclassification, defined as clinically significant prostate cancer of grade group 2 or more, during follow-up according to PSAD, after adjusting for covariates. RESULTS: One hundred twenty-seven (32.6%) patients had mpMRI(-); 72 (18.5%) had PI-RADS 3, 150 (38.6%) PI-RADS 4, and 40 (10.3%) PI-RADS 5 lesions. The rate of reclassification to grade group 2 PCa was 16%, 22%, 31%, and 39% for mpMRI(-) and PI-RADS 3, 4, and 5, respectively, in case of PSAD < 0.10 ng/mL2; 16%, 25%, 36%, and 44%, in case of PSAD 0.10 to 0.19 ng/mL2; and 25%, 42%, 55%, and 67% in case of PSAD ≥ 0.20 ng/mL2. PSAD ≥ 0.20 ng/mL2 (odds ratio [OR], 2.45; P = .007), PI-RADS 3 (OR, 2.47; P = .013), PI-RADS 4 (OR, 2.94; P < .001), and PI-RADS 5 (OR, 3.41; P = .004) were associated with a higher risk of reclassification. CONCLUSION: PSAD ≥ 0.20 ng/mL2 may improve predictive accuracy of mpMRI results for reclassification of patients in AS, whereas PSAD < 0.10 ng/mL2 may help selection of patients at lower risk of harboring clinically significant prostate cancer. However, the risk of reclassification is not negligible at any PSAD cutoff value, also in the case of mpMRI(-).


Subject(s)
Multiparametric Magnetic Resonance Imaging , Prostatic Neoplasms , Follow-Up Studies , Humans , Image-Guided Biopsy , Magnetic Resonance Imaging , Male , Prostate-Specific Antigen , Prostatic Neoplasms/diagnostic imaging , Retrospective Studies , Watchful Waiting
5.
Prostate Int ; 8(4): 167-172, 2020 Dec.
Article in English | MEDLINE | ID: mdl-33425794

ABSTRACT

INTRODUCTION & OBJECTIVES: We tested the role of multiparametric magnetic resonance imaging (mpMRI) in disease reclassification and whether the combination of mpMRI and clinicopathological variables could represent the most accurate approach to predict the risk of reclassification during active surveillance. MATERIALS & METHODS: Three-hundred eighty-nine patients (pts) underwent mpMRI and subsequent confirmatory or follow-up biopsy according to the Prostate Cancer Research International Active Surveillance (PRIAS) protocol. Pts with negative (-) mpMRI underwent systematic random biopsy. Pts with positive (+) mpMRI [Prostate Imaging Reporting and Data System, version 2 (PI-RADS-V2) score ≥3] underwent targeted + systematic random biopsies. Multivariate analyses were used to create three models predicting the probability of reclassification [International Society of Urological Pathology ≥ Grade Group 2 (GG2)]: a basic model including only clinical variables (age, prostate-specific antigen density, and number of positive cores at baseline), an Magnetic resonance imaging (MRI) model including only the PI-RADS score, and a full model including both the previous ones. The predictive accuracy (PA) of each model was quantified using the area under the curve. RESULTS: mpMRI negative (-) was recorded in 127 (32.6%) pts; mpMRI positive (+) was recorded in 262 pts: 72 (18.5%) had PI-RADS 3, 150 (38.6%) PI-RADS 4, and 40 (10.3%) PI-RADS 5 lesions. At a median follow-up of 12 months, 125 pts (32%) were reclassified to GG2 prostate cancer. The rate of reclassification to GG2 prostate cancer was 17%, 35%, 38%, and 52% for mpMRI (-), PI-RADS 3, 4, and 5, respectively (P < 0.001). The PA was 69% and 64% in the basic and MRI models, respectively. The full model had the best PA of 74%: older age (P = 0.023; Odds ratio (OR) = 1.040), prostate-specific antigen density (P = 0.037; OR = 1.324), number of positive cores at baseline (P = 0.001; OR = 1.441), and PI-RADS 3, 4, and 5 (overall P = 0.001; OR = 2.458, 3.007, and 3.898, respectively) were independent predictors of reclassification. CONCLUSIONS: Disease reclassification increased according to the PI-RADS score increase, at confirmatory or follow-up biopsy. However, a no-negligible rate of reclassification was found also in cases of mpMRI (-). The combination of mpMRI and clinicopathological variables still represents the most accurate approach to pts on active surveillance.

6.
Minerva Urol Nefrol ; 71(6): 597-604, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31144491

ABSTRACT

BACKGROUND: The aim of this study was to evaluate intra- and perioperative outcomes of a single high volume open radical prostatectomy (ORP) surgeon, during his learning curve period for robot-assisted radical prostatectomy (RARP) and extended pelvic lymph node dissection (ePLND). METHODS: The study included 264 intermediate-high risk prostate cancer patients, treated by ORP + ePLND or RARP + ePLND, prospectively collected. Descriptive statistics compared clinical and pathological variables between groups. Bivariate (Pearson) correlation analysis assessed the relationship between the number of lymph node (LN) removed, positive surgical margins (PSM), surgical time and the number of procedures performed per group. RESULTS: pT stage and Gleason score (GS) were lower in RARP than in ORP group (both P=0.04), while PSM were more frequent in the RARP group (40% vs. 25%; P=0.02). However, PSM decreased with the increase of RARP procedures. The number of LNs removed was 25 and 22, in RARP and ORP group (P=0.03). However, LN+ rate did not differ between groups (11% vs. 16%; P=0.216). In the RARP group, overall surgical time and ePLND time decreased with the increase of surgical procedures (all P<0.001). CONCLUSIONS: RARP requires significant learning curve to reduce operative room time and obtain PSM comparable to those of an ORP high-volume surgeon. On the contrary, the quality of ePLND during RARP seems to be not related to the number of procedures performed, allowing removal of a number of LNs that is clinically comparable to ORP.


Subject(s)
Lymph Node Excision/methods , Pelvis , Prostatectomy/methods , Robotic Surgical Procedures/methods , Adult , Aged , Aged, 80 and over , Humans , Learning Curve , Male , Margins of Excision , Middle Aged , Neoplasm Grading , Operative Time , Prospective Studies , Prostatic Neoplasms/pathology , Prostatic Neoplasms/surgery , Surgeons , Treatment Outcome
7.
Food Res Int ; 116: 1059-1065, 2019 02.
Article in English | MEDLINE | ID: mdl-30716889

ABSTRACT

In this work, we explored the "deep" seed peanut proteome by using both two dimensional electrophoresis (2-DE)-based analysis run under reducing and non-reducing condition (protein-centric) and LC-MS/MS gel-free proteomic (peptide-centric). The former approach allowed to identify high molecular weight disulfide-linked Ara h 1 and Ara h 3 heteroligomers and Ara h 1 homoligomers linked through covalent bonds other than disulfides. The occurrence of these protein complexes revealed natural interactions between Ara(s) subunits with a possible involvement in the allergenic potential of peanut. The second approach, also referred to as shot-gun proteomics, allowed the identification of 149 gene products, including low-abundance proteins escaped the 2-DE detection. Interestingly, we identified 60 proteins never catalogued previously. The complementary exploitation of two proteomic approaches enabled the access to new relevant information about the complexity of the peanut proteome, with special emphasis to the complement of allergens (allergome).


Subject(s)
Antigens, Plant/isolation & purification , Arachis/chemistry , Chromatography, High Pressure Liquid , Electrophoresis, Gel, Two-Dimensional , Nuts/chemistry , Peanut Hypersensitivity/immunology , Plant Proteins/isolation & purification , Proteomics/methods , Tandem Mass Spectrometry , Antigens, Plant/immunology , Arachis/immunology , Membrane Proteins/isolation & purification , Nuts/immunology , Plant Proteins/immunology , Proteome
8.
Article in German | MEDLINE | ID: mdl-30758636

ABSTRACT

In accordance with Prevention Act law and the associated new prevention mandate of the nursing care funds, more preventive and health promoting activities have been taking place in inpatient care facilities since 2016. The Lübeck model of themed movement activities as a part of the program "Grow older in Balance" of the Federal Center for Health Education was developed as a physical, mental, and socially activating prevention program for elderly people with physical and cognitive impairments.The regional implementation of the Lübeck model in inpatient care facilities, facilities for age-appropriate living, as well as in day-care, serves the Berlin project "Adaptation and implementation of the Lübeck model of themed movement activities in the model region of Pankow - Low threshold exercise offers in nursing facilities for the elderly." This model project of the regional network Qualitätsverbund Netzwerk im Alter - Pankow e. V. (QVNIA e. V.) is presented in the current article.The relevant local structures and cooperation as well as the resulting and program-related requirements for participating actors are highlighted. With the aim of sustainable implementation of physical activity promotion in the nursing setting, a possible implementation path and quality assurance measures are presented.The transfer has shown that the concept of the Lübeck model can also be implemented in metropolitan structures. However, the implementation requires the consideration of the regional framework conditions. A centralized local control with the corresponding structural and procedural expertise as well as quality assurance assets is necessary. Positive experiences in the implementation of the program were acquired during the model phase and recommendations for the continuation and further development can be derived.


Subject(s)
Health Education , Health Promotion , Models, Theoretical , Aged , Berlin , Germany , Humans
9.
Food Res Int ; 115: 360-368, 2019 01.
Article in English | MEDLINE | ID: mdl-30599953

ABSTRACT

In this work, the effects of maturation time and simulated gastrointestinal digestion on the molecular and peptide profiles of "Bresaola Valtellina" were assessed through the foodomics approach, in this case food proteomics and peptidomics combined to other analytical and biological assays, aiming at depicting a holistic food quality. Human digestion of this Italian cured meat product was simulated using an in vitro static protocol and the degree of proteolysis and the in vitro bioactivity of the soluble free compounds in the digestates were evaluated by biochemical assays, e.g. SDS-PAGE, size exclusion HPLC, HPLC/MS, 1H NMR, enzymatic and antioxidant activities. The obtained results demonstrated that in vitro gastrointestinal digestion contributed to a considerable release of myofibrillar proteins by the muscle tissue. Data from SDS-PAGE, peptidomic and size exclusion HPLC assays showed that the in vitro digestion largely degraded proteins of muscle tissue to peptides smaller than 250 Da. The released peptides were likely responsible for the inhibitory activity on amylolytic enzymes and for the antioxidant properties elicited by the gastric digestates of Bresaola. Overall, the results demonstrated the negligible role of ripening in making meat proteins more bioaccessible, whereas they confirmed the highly in vitro digestibility of meat proteins from Bresaola. This study represents a new approach merging proteomics and foodomics to evaluate the effect of ripening and in vitro digestion on the bioactivity and bioaccessibility of proteins and peptides of meat products.


Subject(s)
Digestion , Meat Products/analysis , Peptides/analysis , Proteins/analysis , Antioxidants/analysis , Chromatography, High Pressure Liquid , Electrophoresis, Polyacrylamide Gel , Food Analysis , Food Quality , Gastrointestinal Tract/chemistry , Humans , Italy , Mass Spectrometry , Peptides/chemistry , Proteolysis , Proteomics
10.
Food Res Int ; 115: 562-571, 2019 01.
Article in English | MEDLINE | ID: mdl-30599980

ABSTRACT

Hemp (Cannabis sativa L.), traditionally cultivated for industrial use and harvested for fibers and seeds, has raised much interest as a sustainable crop in the last years. Recently, hemp seeds and derived oil have started to be used in a variety of food products. Hemp-based food products are considered less allergenic than those from other edible seeds, although this statement has never been experimentally verified. In this study high purity grade hemp flour (HF) and hemp protein isolate (HPI) were obtained through a fast and cheap process starting from defatted hemp cakes, a residue of hempseed oil extraction. HPI resulted enriched at nearly 86% protein, mainly constituted by the storage protein edestin (accounting for 70% total protein). In vitro protein digestibility was determined using a static model of gastrointestinal digestion (GID), which included a final step with purified brush border membrane (BBM) enzyme preparations. HF and HPI showed a high degree of digestibility. The survival of potential bioactive and/or allergenic peptide sequences in digests was investigated by peptidomic analysis. Only a limited number of sequences survived GID. Among them, fragments from 12 seed proteins. These fragments were precursors of sequences with potential bioactive peptides, which might justify the bioactivity of HPI hydrolysates, reported in previous studies. More importantly, all known hemp allergens, including the major thaumatin-like protein and LTP, were entirely eliminated by the HPI production process, neither fragments of the proteins were present after GID. These data support the use of HPI as an ingredient for hypoallergenic foods.


Subject(s)
Allergens/analysis , Cannabis/chemistry , Digestion , Plant Proteins/analysis , Flour , Food Hypersensitivity , Peptides/analysis , Peptides/immunology , Plant Proteins/immunology , Plant Proteins/metabolism , Protein Hydrolysates/analysis , Proteomics , Seeds/chemistry
11.
Food Chem ; 277: 195-204, 2019 Mar 30.
Article in English | MEDLINE | ID: mdl-30502135

ABSTRACT

The need of controlling illegal addition of water buffalo (WB) milk from foreign countries to the Italian counterpart devoted to the production of Protected Denomination of Origin (PDO) Mozzarella di Bufala Campana (MBC) cheese has promoted the development of simple, fast and cheap isoelectrofocusing (IEF) methods for evaluating the nature of the raw material to be used according to a high-throughput sample multiplexing format, avoiding the use of dedicated mass spectrometry-based procedures. Thus, combined proteomic methods were here integrated with optimized western blotting protocols in solving the complex IEF pattern of casein (CN) mixtures observed when Italian and foreign WB milk are mixed together. Identification of internally deleted αs1-CN hepta-phosphorylated species as well as of still unknown ß-CN A hexa-phosphorylated and N-terminally-nicked ß-CN A phosphorylated forms present uniquely in foreign WB milk samples, allowed recognizing these molecules as adulteration markers to be assayed in combined IEF-immunoblotting procedures; the latter ones showing optimal migration characteristics to be used in routine assays. A linear relationship between detected area of specific immunorecognized gel bands and percentage of international WB milk added to the Italian counterpart was verified, demonstrating that this method has an adulteration detection limit close to 3% v/v. Based on these results, this analytical procedure is here proposed as optimal one for evaluating the authenticity of PDO MBC cheese products.


Subject(s)
Buffaloes , Caseins/chemistry , Caseins/metabolism , Food Quality , Fraud/prevention & control , Milk/chemistry , Animals , Binding Sites , Biomarkers/chemistry , Biomarkers/metabolism , Immunoblotting , Isoelectric Focusing , Phosphorylation
12.
Front Surg ; 5: 52, 2018.
Article in English | MEDLINE | ID: mdl-30246012

ABSTRACT

Objectives: To evaluate the frequency and distribution of pelvic nodes metastases, in intermediate-high risk prostate cancer (PCa) patients (pts), who underwent open radical prostatectomy (ORP) and superextended pelvic lymph node dissection (sePLND). Patients and Methods: We retrospectively evaluated 630 consecutive pts with clinically localized, intermediate-high risk PCa, treated with ORP and sePLND from 2009 to 2016 at a single institution. The sePLND always removed all nodal/fibro-fatty tissue of the internal iliac, external iliac, obturator, common iliac, and presacral regions. Results: Positive lymph nodes (LN+) were found in 133 pts (21.1%). The median number of removed nodes and LN+ was 25 and 1, respectively. LN+ were found in 64 (48.1%), 58 (43.6%), 53 (39.8%), 16 (12%), and 20 (15%) pts and were present as a single site in 27 (20.3%), 22 (16.5%), 20 (15%), 0, and 6 (4.5%) cases in the internal iliac, external iliac, obturator, common iliac, and presacral chain, respectively. An ePLND would have correctly staged 127 (95%) pts but removed all LN+ in only 97 (73%) pts. Presacral nodes harbored LN+ in 20 patients. Among them, 18 were high-risk patients. Moreover, all but 1 pts with common iliac LN+ were in high risk group. Conclusions: These results suggest that removal of presacral and common iliac nodes could be omitted in intermediate risk pts. However, a PLND limited to external iliac, obturator, and internal iliac region may be adequate for nodal staging purpose, but not enough accurate if we aim to remove all possible site of LN+ in high risk pts.

13.
Food Chem ; 230: 482-490, 2017 Sep 01.
Article in English | MEDLINE | ID: mdl-28407938

ABSTRACT

The European reference method (ERM) recognises the fraudulent addition of bovine (B) milk in water buffalo (WB) milk/dairy products based on concomitant isoelectric focusing (IEF) detection of B γ2- and γ3-CN fragments after corresponding plasminolysis. We here used proteomics to characterise false positive results occurring in the ERM as being due to WB ß-CN(f100-209), which is also formed after plasminolysis of genuine WB milk/dairy products and comigrates in IEF with B γ2-CN. These ERM limitations were overcome by a dedicated proteomic procedure based on loading of B/WB milk/cheese CN extracts on a hydroxyapatite column, in situ trypsinolysis and elution of B ß-CN(f1-25)4P and WB ß-CN(f1-28)4P proteotypic peptides. Based on their similar ionisation properties and resolution in MALDI-TOF-MS, these phosphopeptides were identified as suitable markers for detection of B material in WB milk/dairy products to a detection limit of 0.8% v/v. This proteomic procedure is here proposed as integrative/alternative to the ERM.


Subject(s)
Caseins/chemistry , Cheese/analysis , Milk/chemistry , Animals , Buffaloes , Cattle , Proteomics
14.
Anal Bioanal Chem ; 408(20): 5609-21, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27299776

ABSTRACT

Water buffalo (WB) casein (CN) and curd samples from indigenous Italian and international breeds were examined with the objective of identifying signature peptides that could function as an indicator to determine the origin of their milk products. CN in complex mixtures were digested with trypsin, and peptide fragments were subsequently identified by matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry (MALDI-TOF MS). The unique presence of a ß-CN A variant and an internally deleted αs1-CN (f35-42) variant in international WB milk samples was ascertained by identifying signature tryptic peptides from either dephosphorylated or native CN. Four signature unphosphorylated peptides derived from ß-CN A, i.e. (f49-68) Asn(68) (2223.6 Da), (f1-28) Ser(10) (3169.4 Da), (f1-29) Ser(10) (3297.4 Da) and (f33-48) Thr(41) (1982 Da) and two from αs1-CN (f35-42) deleted fragments, i.e. (f23-34) Met(31) (1415.7 Da) and (f43-58) Val(44) (1752.7 Da), were identified. Two signature casein phosphopeptides (CPPs), i.e. ß-CN (f1-28) 4P (3489.1 Da) and ß-CN (f33-48) 1P (2062.0 Da), were identified in the tryptic hydrolysate of native casein or curd and cheese samples using in-batch hydroxyapatite (HA) chromatography. All these fragments functioned as analytical surrogates of two αs1- and ß-casein variants that specifically occur in the milk of international WB breeds. Furthermore, the bovine peptide ß-CN (f1-28) 4P had a distinct and lower molecular mass compared with the WB counterpart and functioned as a species-specific marker for all breeds of WB. Advantages of this analytical approach are that (i) peptides are easier to separate than proteins, (ii) signature peptide probes originating from specific casein variants allow for the targeting of all international WB milk, curd and cheese samples and (iii) bovine and WB casein in mixtures can be simultaneously determined in protected designation of origin (PDO) "Mozzarella di Bufala Campana" cheese. This analytical method enabled the specific detection of international WB and bovine casein with a sensitivity threshold of 2 and 0.78 %, respectively. Graphical Abstract Monitoring of prototypic tryptic CPPs by MALDI-TOF analysis in Mediterranean (A), Romanian (B), Indian (C), Polish (D) and Canadian (E) curd samples to guarantee the authenticity of the PDO "Mozzarella di Bufala Campana" cheese.


Subject(s)
Caseins/chemistry , Cheese/analysis , Food Analysis/methods , Food Contamination/analysis , Milk/chemistry , Peptide Mapping/methods , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization/methods , Animals , Buffaloes , Caseins/analysis , Cattle , Cheese/classification , Internationality , Italy , Milk/classification
15.
Radiat Environ Biophys ; 54(2): 155-66, 2015 May.
Article in English | MEDLINE | ID: mdl-25600561

ABSTRACT

The long-term goal to integrate laser-based particle accelerators into radiotherapy clinics not only requires technological development of high-intensity lasers and new techniques for beam detection and dose delivery, but also characterization of the biological consequences of this new particle beam quality, i.e. ultra-short, ultra-intense pulses. In the present work, we describe successful in vivo experiments with laser-driven electron pulses by utilization of a small tumour model on the mouse ear for the human squamous cell carcinoma model FaDu. The already established in vitro irradiation technology at the laser system JETI was further enhanced for 3D tumour irradiation in vivo in terms of beam transport, beam monitoring, dose delivery and dosimetry in order to precisely apply a prescribed dose to each tumour in full-scale radiobiological experiments. Tumour growth delay was determined after irradiation with doses of 3 and 6 Gy by laser-accelerated electrons. Reference irradiation was performed with continuous electron beams at a clinical linear accelerator in order to both validate the dedicated dosimetry employed for laser-accelerated JETI electrons and above all review the biological results. No significant difference in radiation-induced tumour growth delay was revealed for the two investigated electron beams. These data provide evidence that the ultra-high dose rate generated by laser acceleration does not impact the biological effectiveness of the particles.


Subject(s)
Electrons/therapeutic use , Lasers , Particle Accelerators , Radiotherapy/instrumentation , Animals , Carcinoma, Squamous Cell/pathology , Carcinoma, Squamous Cell/radiotherapy , Cell Line, Tumor , Cell Proliferation/radiation effects , Cell Transformation, Neoplastic , Dose-Response Relationship, Radiation , Female , Humans , Male , Mice , Radiometry
16.
Nat Commun ; 4: 2421, 2013.
Article in English | MEDLINE | ID: mdl-24026068

ABSTRACT

Laser-plasma particle accelerators could provide more compact sources of high-energy radiation than conventional accelerators. Moreover, because they deliver radiation in femtosecond pulses, they could improve the time resolution of X-ray absorption techniques. Here we show that we can measure and control the polarization of ultra-short, broad-band keV photon pulses emitted from a laser-plasma-based betatron source. The electron trajectories and hence the polarization of the emitted X-rays are experimentally controlled by the pulse-front tilt of the driving laser pulses. Particle-in-cell simulations show that an asymmetric plasma wave can be driven by a tilted pulse front and a non-symmetric intensity distribution of the focal spot. Both lead to a notable off-axis electron injection followed by collective electron-betatron oscillations. We expect that our method for an all-optical steering is not only useful for plasma-based X-ray sources but also has significance for future laser-based particle accelerators.

17.
Food Funct ; 4(1): 144-52, 2013 Jan.
Article in English | MEDLINE | ID: mdl-23104098

ABSTRACT

The use of microalgae as a food source is still poorly developed because of the technical difficulties related to their cultivation and the limited knowledge about their chemical composition and nutritional value. The unicellular red microalga Galdieria sulphuraria has a very high daily productivity and its cultivation under acidic conditions avoided any bacterial contamination. G. sulphuraria can be cultured under autotrophic and heterotrophic conditions: in this study a screening of 43 strains showed that in the latter case a duplication of biomass production was obtained. The proximate composition (protein, carbohydrates, fiber and lipids), the micronutrient content (carotenoids, phycobiliproteins, chlorophylls and vitamins) together with the antioxidant activity of the biomass produced by a selected strain of G. sulphuraria under both cultivation conditions were determined. Results showed that the material is rich in proteins (26-32%) and polysaccharides (63-69%) and poor in lipids. Under heterotrophic cultivation conditions, the lipid moiety mainly contained monounsaturated fatty acids. Among micronutrients, some B group vitamins are present, beta-carotene is the main carotenoid and phycobiliproteins are present under both cultivating conditions. G. sulphuraria proteins are strictly associated with polysaccharide components and therefore not digestible. In the second part of the work, an extraction protocol using Viscozyme L, a commercial enzymatic preparation containing a mixture of polysaccharidases, was developed which made G. sulphuraria proteins a good substrate for human gastrointestinal enzymes. All in all, the data suggested that G. sulphuraria biomass has a potential use as food ingredients both for protein-rich or insoluble dietary fibre-rich applications. The low concentration of lipids and the absence of green color make this microalgae source particularly useful for the addition to many food preparations.


Subject(s)
Food Analysis/methods , Microalgae/chemistry , Microalgae/metabolism , Nutritive Value , Humans , Phenols , Plant Extracts , Plant Proteins/chemistry , Plant Proteins/metabolism
18.
Electrophoresis ; 33(15): 2337-44, 2012 Aug.
Article in English | MEDLINE | ID: mdl-22887154

ABSTRACT

A genetic survey on three autochthonous goat breeds reared in Italy was carried out by a proteomic approach. This methodology, further to providing the phenotypic frequency of identified α(s1) genetic variants, allowed to determine (i) the additional constitutive presence of a non-allelic 'α(s1) -casein (CN) F like' protein in goat 'strong' α(s1) variants; (ii) an α(s1) -CN B(2) like protein, expressed at very low quantitative level, in goat 'weak' α(s1) -CN variants, and, as main focus; (iii) the occurrence of a new α(s1) -CN D(1) variant characterised by the lack of α(s1) (f59-69) sequence otherwise encoded by exon 9 in goat α(s1) B(2) reference. The same exon skipping event had been identified since 1990, as responsible of the 'weak quantitative class' of α(s1) -CN D variant (0.6 g/L), while the new α(s1) -CN D(1,) has been 'quantitatively' classified as an 'intermediate' variant, since 1.8 g/L per allele was assessed in the milk.


Subject(s)
Caseins/genetics , Goats/genetics , Amino Acid Sequence , Animals , Caseins/analysis , Caseins/chemistry , Chromatography, High Pressure Liquid , Chromatography, Reverse-Phase , Electrophoresis, Gel, Two-Dimensional , Exons , Immunoblotting , Mass Spectrometry/methods , Polymorphism, Genetic , Proteomics
19.
J Agric Food Chem ; 60(32): 8044-50, 2012 Aug 15.
Article in English | MEDLINE | ID: mdl-22809470

ABSTRACT

A clear picture of the protein rearrangement in milk following UHT-treatment was drawn by a comparative analysis of the pH 4.6 soluble protein fraction (SPF) and the pH 4.6 insoluble protein fraction (IPF) recovered from raw and UHT-treated milk samples. The two protein fractions were analyzed by mono- or bidimensional gel electrophoresis under reducing and nonreducing conditions, and protein bands were identified by specific immunostaining. Results showed that bovine serum albumin, ß-lactoglobulin, and, to a lesser extent, α-lactalbumin coprecipitated with caseins in UHT-treated milk samples at pH 4.6. These proteins were almost exclusively involved in high molecular weight aggregates held together by disulfide bonds. Partition of α-lactalbumin and bovine serum albumin in the protein fractions obtained upon acidification of milk at pH 4.6 was evaluated by competitive immunoassays. The ELISA-based results suggested the possibility of using pH 4.6 insoluble α-lactalbumin and bovine serum albumin, in addition to pH 4.6 insoluble ß-lactoglobulin, as indicators of the intensity of the heat treatment applied to milk.


Subject(s)
Food Handling/methods , Hot Temperature , Milk Proteins/analysis , Milk/chemistry , Animals , Caseins/analysis , Hydrogen-Ion Concentration , Lactalbumin/analysis , Lactoglobulins/analysis , Milk Proteins/chemistry , Serum Albumin, Bovine/analysis , Solubility
20.
J Radiat Res ; 53(3): 395-403, 2012.
Article in English | MEDLINE | ID: mdl-22739009

ABSTRACT

The notable progress in laser particle acceleration technology promises potential medical application in cancer therapy through compact and cost effective laser devices that are suitable for already existing clinics. Previously, consequences on the radiobiological response by laser driven particle beams characterised by an ultra high peak dose rate have to be investigated. Therefore, tumour and non-malignant cells were irradiated with pulsed laser accelerated electrons at the JETI facility for the comparison with continuous electrons of a conventional therapy LINAC. Dose response curves were measured for the biological endpoints clonogenic survival and residual DNA double strand breaks. The overall results show no significant differences in radiobiological response for in vitro cell experiments between laser accelerated pulsed and clinical used electron beams. These first systematic in vitro cell response studies with precise dosimetry to laser driven electron beams represent a first step toward the long term aim of the application of laser accelerated particles in radiotherapy.


Subject(s)
Electrons/therapeutic use , Neoplasms/radiotherapy , Cell Line , Cell Line, Tumor , Cell Survival/radiation effects , DNA Breaks, Double-Stranded , Dose-Response Relationship, Radiation , Equipment Design , Humans , Laser Therapy/instrumentation , Neoplasms/metabolism , Neoplasms/pathology , Particle Accelerators/instrumentation , Relative Biological Effectiveness , Tumor Stem Cell Assay
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