Serum metabolome differences associated with subclinical intramammary infection caused by Streptococcus agalactiae and Prototheca spp. in multiparous dairy cows.

Mastitis is one of the most significant diseases in dairy cows and causes several economic losses. Somatic cell count (SCC) is often used as an indirect diagnostic tool for mastitis, especially for subclinical mastitis (SCM) where no symptoms or signs can be detected. Streptococcus agalactiae is one of the main causes of contagious mastitis, and Prototheca spp. is an alga-inducing environmental mastitis that is not always correlated with increased milk SCC. The aim of this study was to evaluate the changes in the metabolomic profile of blood in relation to subclinical intramammary infection (IMI) in dairy cows. In addition, differences resulting from the etiologic agent causing mastitis were also considered. Forty Holstein-Friesian dairy cows in mid and late lactation were enrolled in this cross-sectional design study. Based on the bacteriological examination of milk, the animals were divided into 3 groups: group CTR (control group; n = 16), group A (affected by SCM with IMI caused by Strep. agalactiae; n = 17), and group P (affected by SCM with IMI caused by Prototheca spp.; n = 7). Blood samples from the jugular vein were collected in tubes containing clot activator; the serum aliquot was stored until metabolomic analysis by 1H-nuclear magnetic resonance spectroscopy. Statistical analysis was conducted by fitting a linear model with the group as the fixed effect and SCC as the covariate. Forty-two metabolites were identified, and among them 10 were significantly different among groups. Groups A and P showed greater levels of His and lactose and lower levels of acetate, Asn, and dimethylamine compared with group CTR. Group A showed high levels of Val, and group P showed high levels of Cit and methylguanidine, as well as lower levels of 3-hydroxybutyrate, acetone, allantoin, carnitine, citrate, and ethanol. These metabolites were related to ruminal fermentations, energy metabolism, urea synthesis and metabolism, immune and inflammatory response, and mammary gland permeability. These results suggest systemic involvement with subclinical IMI and that the metabolic profile of animals with SCM undergoes changes related to the etiological agent of mastitis.

Serum metabolomics assessment of etiological processes predisposing ketosis in water buffalo during early lactation.

Metabolic disorders as ketosis are manifestations of the animal's inability to manage the increase in energy requirement during early lactation. Generally, buffaloes show a different response to higher metabolic demands than other ruminants with a lower incidence of metabolic problems, although ketosis is one of the major diseases that may decrease the productivity in buffaloes. The aim of this study was to characterize the metabolic profile of Mediterranean buffaloes (MB) associated with 2 different levels of ß-hydroxybutyrate (BHB). Sixty-two MB within 50 days in milk (DIM) were enrolled and divided into 2 groups according to serum BHB concentration: healthy group (37 MB; BHB <0.70 mmol/L; body condition score: 5.00; parity: 3.78; and DIM: 30.70) and group at risk of hyperketonemia (25 MB; BHB ≥0.70 mmol/L; body condition score: 4.50; parity: 3.76; and DIM: 33.20). The statistical analysis was conducted by one-way ANOVA and unpaired 2-sample Wilcoxon tests. Fifty-seven metabolites were identified and among them, 12 were significant or tended to be significant. These metabolites were related to different metabolic changes such as mobilization of body resources, ruminal fermentations, urea cycle, thyroid hormone synthesis, inflammation, and oxidative stress status. These findings are suggestive of metabolic changes related to subclinical ketosis status that should be further investigated to better characterize this disease in the MB.

Canonical and uncanonical pathogenic germline variants in colorectal cancer patients by next-generation sequencing in a European referral center.

BACKGROUND: Despite increasing use of next-generation sequencing (NGS), data concerning the gain in germline pathogenic variants (PVs) remain scanty, especially with respect to uncanonical ones. We aimed to verify the impact of different cancer predisposition genes (CPGs) on colorectal cancer (CRC) in patients referred for genetic evaluation. MATERIALS AND METHODS: We enrolled for NGS, by Illumina TruSight Cancer panel comprising 94 CPGs, 190 consecutive subjects referred for microsatellite instability (MSI) CRC, polyposis, and/or family history. RESULTS: Overall, 51 (26.8%) subjects carried 64 PVs; PVs coexisted in 4 (7.8%) carriers. PVs in mismatch repair (MMR) genes accounted for one-third of variant burden (31.3%). Four Lynch syndrome patients (20%) harbored additional PVs (HOXB13, CHEK2, BRCA1, NF1 plus BRIP1); such multiple PVs occurred only in subjects with PVs in mismatch syndrome genes (4/20 versus 0/31; P = 0.02). Five of 22 (22.7%) patients with MSI cancers but wild-type MMR genes harbored PVs in unconventional genes (FANCL, FANCA, ATM, PTCH1, BAP1). In 10/63 patients (15.9%) with microsatellite stable CRC, 6 had MUTYH PVs (2 being homozygous) and 4 exhibited uncanonical PVs (BRCA2, BRIP1, MC1R, ATM). In polyposis, we detected PVs in 13 (25.5%) cases: 5 (9.8%) in APC, 6 (11.8%) with biallelic PVs in MUTYH, and 2 (3.9%) in uncanonical genes (FANCM, XPC). In subjects tested for family history only, we detected two carriers (18.2%) with PVs (ATM, MUTYH). CONCLUSION: Uncanonical variants may account for up to one-third of PVs, underlining the urgent need of consensus on clinical advice for incidental findings in cancer-predisposing genes not related to patient phenotype.

Breast muscle and plasma metabolomics profile of broiler chickens exposed to chronic heat stress conditions.

Understanding the variations of muscle and plasma metabolites in response to high environmental temperature can provide important information on the molecular mechanisms related to body energy homeostasis in heat-stressed broiler chickens. In this study, we investigated the effect of chronic heat stress conditions on the breast muscle (Pectoralis major) and plasma metabolomics profile of broiler chickens by means of an innovative, high-throughput analytical approach such as the proton nuclear magnetic resonance (1H NMR) spectrometry. A total of 300 Ross 308 male chicks were split into two experimental groups and raised in either thermoneutral conditions for the entire rearing cycle (0-41 days) (TNT group; six replicates of 25 birds/each) or exposed to chronic heat stress conditions (30 °C for 24 h/day) from 35 to 41 days (CHS group; six replicates of 25 birds/each). At processing (41 days), plasma and breast muscle samples were obtained from 12 birds/experimental group and then subjected to 1H NMR analysis. The reduction of BW and feed intake as well as the increase in rectal temperature and heterophil: lymphocyte ratio confirmed that our experimental model was able to stimulate a thermal stress response without significantly affecting mortality. The 1H NMR analysis revealed that a total of 26 and 19 molecules, mostly related to energy and protein metabolism as well as antioxidant response, showed significantly different concentrations respectively in the breast muscle and plasma in response to the thermal challenge. In conclusion, the results obtained in this study indicated that chronic heat stress significantly modulates the breast muscle and plasma metabolome in fast-growing broiler chickens, allowing to delineate potential metabolic changes that can have important implications in terms of body energy homeostasis, growth performance and product quality.

Effect of broiler breast abnormality and freezing on meat quality and metabolites assessed by 1H-NMR spectroscopy.

The present study aimed at investigating the impact of the current growth-related abnormalities (White-Striping-WS, Wooden Breast-WB, and Spaghetti Meat-SM) affecting broilers Pectoralis major muscles on the main quality traits, the oxidative stability of both the lipid and protein fraction as well as the water mobility assessed in fresh and frozen/thawed meat. In addition, Nuclear Magnetic Resonance spectroscopy (1H-NMR) was applied to quantify free amino acids, histidine-containing dipeptides and metabolites involved in energy-generating pathways. Overall, the occurrence of WS, WB, and SM abnormalities remarkably affected the quality traits (pH, color, and water holding capacity) and oxidative stability of the meat, with the WB condition leading to the most detrimental effects. However, overall, freezing and subsequent thawing only partially worsened the aforementioned traits. Significant variations in free amino acids and histidine-containing dipeptides were found between abnormal muscles and their unaffected counterpart by 1H-NMR spectroscopy and, aside from the occurrence of muscular defects, their content was remarkably reduced in frozen/thawed meat. The findings obtained by analyzing the metabolites through 1H-NMR spectroscopy allowed to advance the knowledge concerning the impact of freezing and subsequent thawing on meat quality traits and provided useful information concerning the underlying mechanisms responsible for the development of WS, WB, and SM abnormalities in broilers.

Gaping of pectoralis minor muscles: magnitude and characterization of an emerging quality issue in broilers.

Recently, a certain number of broiler abattoirs located in different Countries around the World have signaled an emerging quality issue termed "gaping" because of the separation of the fiber bundles affecting the external portion of the bipinnate pectoralis minor muscle. Thus, after defining the criteria to classify the muscles as Normal (NORM), Moderate (MOD), or Severe (SEV) cases, the incidence of gaping under commercial conditions was assessed on a total of 8,600 P. minor obtained from broiler chickens belonging to 43 flocks during a 6-mo period. Then, a total of 180 P. minor were selected based on previously defined criteria to evaluate the main quality traits (pH, color, water-holding/-binding capacity and tenderness), proximate composition, water mobility, and thermal properties as well as metabolic profile through 1H-Nuclear Magnetic Resonance spectroscopy. The average incidence of gaping defect was found to be 16.8% (8.8 and 8.0% MOD and SEV cases, respectively). As for the main quality traits, a reduction in ultimate pH was observed as the severity of the gaping defect increased, with SEV muscles displaying significantly lower values in comparison with NORM (5.96 vs. 6.02; P < 0.01), while MOD showed intermediate values (5.99). Concurrently, if compared with their NORM counterpart, MOD and SEV exhibited higher lightness (53.6 and 54.2 vs. 51.8; P < 0.01) coupled with higher (P < 0.05) cooking losses and longer (P < 0.05) transversal relaxation time of extra-myofibrillar water fraction. Overall, no significant differences were found concerning proximate composition and thermal properties. With regard to the metabolic profile, a significantly lower (P < 0.001) glutamine concentration was found in MOD and SEV muscles that, concurrently, revealed significant (P < 0.05) variations in the metabolites involved in energy-generating pathways. Overall, these findings evidenced that the gaping defect affecting broilers' P. minor muscles have strong similarities with the pale-soft-exudative condition previously described in poultry and likely results from the biochemical processes taking place during the post-mortem conversion of muscle to meat.

Impact of treatments on fecal microbiota and fecal metabolome in symptomatic uncomplicated diverticular disease of the colon: a pilot study.

Symptomatic uncomplicated diverticular disease (SUDD) affects 50% of people having diverticulosis. We performed a pilot study assessing the effect of current treatments on fecal microbiota and metabolome in SUDD. Thirteen consecutive females with SUDD were treated with a 2-week therapeutic trial of 30 g/day fiber supplementation (3 patients), 1.6 g/day of mesalazine (3 patients), 900 billion/day of probiotic mixture VivoMixx® (3 patients), or 800 mg/day of rifaximin (4 patients). Stool samples were collected at entry (T0), at the end of the 2-week therapeutic course (T1), and 30 (T2) and 60 days (T3) after the end of the therapeutic course. Real-time PCR quantified targeted microorganisms. Fecal metabolome patterns were studied by high-resolution proton NMR spectroscopy. At cumulative analysis, symptoms significantly decreased at each time point during follow-up (p less than 0.0001), and only left-lower quadrant pain increased again at T3. The overall bacterial quantity was not altered by the treatments. The amount of Akkermansia muciniphila species was significantly reduced at T1 (p=0.017) and at T2 (p=0.026), while at T3 the reduction was not significant in comparison to enrollment (p=0.090). Fecal molecular profile showed significant changes at T1 and T2, while at T3 it became similar to that of T0. Differences were found for 18 of the quantified molecules (tryptophan, phenylalanine, tyrosine, 4-hydroxyphenylacetate, urocanate, X-6.363, X-5.779, uridylate, galactose, X-4.197, threonine, sarcosine, methionine, 2-oxoisocaproate, 5-aminolevulinate, alanine, leucine, valerate). Metabolome and microbiota changed in patients with SUDD under treatment, confirming a possible role of dysbiosis/dysmetabolome in the pathology.

Effect of white striping on turkey breast meat quality.

In the past decades, the intense selection practices carried out in order to develop fast growing and high breast-yield turkey hybrids profoundly modified the muscle physiology leading to the development of growth-related alterations and muscular abnormalities. White striations of variable thickness have been particularly observed on the ventral surface of Pectoralis major muscle belonging from heavy male turkeys since several years. However, although the effects of white striping (WS) have been extensively studied on broilers, this condition was not considered as a main quality issue by both turkey producers and meat industry. Thus, this study aimed at evaluating whether the occurrence of WS in heavy male turkeys affects the quality traits and technological properties of meat to the same extent previously observed for broilers. In two replications, 72 Pectoralis major muscles were classified as: normal (NORM), moderate WS (MOD) and severe WS (SEV) cases. The whole muscle was weighed and cut in order to assess colour, ultimate pH, water holding (drip and cooking losses) and binding (marinade uptake) capacities, NMR relaxation properties, shear force as well as proximate composition of meat. The Pectoralis major muscles affected by WS (both moderate and severe cases) exhibited a one-fifth increased weight in comparison with their NORM counterpart. However, the occurrence of WS only partially affected the proximate composition of the meat. In detail, although moisture, collagen and protein contents did not differ among the groups, if compared with NORM, higher lipid levels were found in SEV muscles, whereas MOD had intermediate values. On the other hand, both MOD and SEV exhibited lower ash content. Despite these variations in proximate composition, both water holding and binding capacities of turkey breast meat were not affected by WS. Indeed, quality traits of raw (pH, colour, cooking losses and shear force) and marinated (uptake, cooking losses and shear force) meat as well as water distribution within the muscle tissue did not differ between NORM and WS cases. Overall, if compared with broilers, WS only marginally affected quality traits of turkey breast meat. It might thus be hypothesised a diverse specie-specific physiological response to the pressure in muscle tissue induced by the selection in turkeys that, although analogously led to the occurrence of WS, results in limited effects on meat quality.

Implications of white striping and spaghetti meat abnormalities on meat quality and histological features in broilers.

During the past few years, there has been an increasing prevalence of broiler breast muscle abnormalities, such as white striping (WS) and wooden breast conditions. More recently, a new muscular abnormality termed as spaghetti meat (SM) because of the altered structural integrity of the Pectoralis major muscle often associated with WS has emerged. Thus, this study aimed at evaluating the effects of WS and SM conditions, occurring alone or combined within the same P. major muscle, on meat quality traits and muscle histology. In two replications, 96 P. major muscles were classified into four classes: normal (N), WS, SM and WS/SM. The whole fillet was used for weight assessment and morphometric measurements, then each sample was cut in order to separate the superficial layer from the deep one and used to evaluate proximate composition, histological features, nuclear magnetic resonance relaxation times, functional properties and both myofibrillar and sarcoplasmic proteins profile. Fillets affected by WS and SM abnormalities exhibited higher weights and increased thickness and length. SM condition was associated with a relevant decrease in protein content coupled with a significant increase in moisture level, whereas fat content was affected only by the simultaneous presence of WS. Histological evaluations revealed that abnormal samples were characterized by several degenerative aspects that almost completely concerned the superficial layer of the fillets. White striped fillets exhibited necrosis and lysis of fibers, fibrosis, lipidosis, loss of cross striation and vacuolar degeneration. Moreover, SM samples were characterized by poor fiber uniformity and a progressive rarefaction of the endo- and peri-mysial connective tissue, whereas WS/SM fillets showed intermediate histological features. Nuclear magnetic resonance relaxation analysis revealed a higher proportion of extra-myofibrillar water in the superficial section of all the abnormal fillets, especially in SM samples, which consequently led to a reduction of the water holding capacity of meat. As for functional properties, abnormal fillets exhibited a lower protein solubility and higher ultimate pH values on both the superficial and deep sections. Although abnormal fillets exhibited higher yellowness values, no relevant effect on meat color was observed. The occurrence of WS and SM abnormalities led to increased carbonylation levels and more intense proteolytic processes. Overall, muscle abnormalities mainly affect the superficial layer of P. major muscle and particularly the occurrence of SM myopathy seems to implicate a more pronounced modification of meat quality traits than the mere presence of WS.

A first step towards a consensus static in vitro model for simulating full-term infant digestion.

In vitro alternatives to clinical trials are used for studying human food digestion. For simulating infant digestion, only a few models, lacking physiological relevance, are available. Thanks to an extensive literature review of the in vivo infant digestive conditions, a gastrointestinal static in vitro model was developed for infants born at term and aged 28days. The model was applied to the digestion of a commercial infant formula. Kinetics of digestion, as well as the structural evolution, were compared with those obtained while submitting the same formula to the adult international consensus protocol of in vitro static digestion. The kinetics of proteolysis and lipolysis differed according to the physiological stage resulting mainly from the reduced level of enzymes and bile salts, as well as the higher gastric pH in the infant model. This in vitro static model of infant digestion is of interest for scientists, food or pharmaceutical manufacturers.

Vaginal microbiome and metabolome highlight specific signatures of bacterial vaginosis.

In this study, we sought to find novel bacterial and metabolic hallmarks for bacterial vaginosis (BV). We studied the vaginal microbiome and metabolome of vaginal fluids from BV-affected patients (n = 43) and healthy controls (n = 37) by means of an integrated approach based on quantitative polymerase chain reaction (qPCR) and proton nuclear magnetic resonance ((1)H-NMR). The correlations between the clinical condition and vaginal bacterial communities were investigated by principal component analysis (PCA). To define the metabolomics signatures of BV, 100 discriminant analysis by projection on latent structure (PLS-DA) models were calculated. Bacterial signatures distinguishing the health condition and BV were identified by qPCR. Lactobacillus crispatus strongly featured the healthy vagina, while increased concentrations of Prevotella, Atopobium and Mycoplasma hominis specifically marked the infection. (1)H-NMR analysis has led to the identification and quantification of 17 previously unreported molecules. BV was associated with changes in the concentration of metabolites belonging to the families of amines, organic acids, short chain fatty acids, amino acids, nitrogenous bases and monosaccharides. In particular, maltose, kynurenine and NAD(+) primarily characterised the healthy status, while nicotinate, malonate and acetate were the best metabolic hallmarks of BV. This study helps to better understand the role of the vaginal microbiota and metabolome in the development of BV infection. We propose a molecular approach for the diagnosis of BV based on quantitative detection in the vaginal fluids of Atopobium, Prevotella and M. hominis, and nicotinate, malonate and acetate by combining qPCR and (1)H-NMR.

Molecular heterogeneity and prognostic implications of synchronous advanced colorectal neoplasia.

BACKGROUND: It is uncertain whether synchronous colorectal cancers (S-CRCs) preferentially develop through widespread DNA methylation and whether they have a prognosis worse than solitary CRC. As tumours with microsatellite instability (MSI) may confound the effect of S-CRC methylation on outcome, we addressed this issue in a series of CRC characterised by BRAF and MS status. METHODS: Demographics, clinicopathological records and disease-specific survival (DSS) were assessed in 881 consecutively resected CRC undergoing complete colonoscopy. All tumours were typed for BRAF(c.1799T>A) mutation and MS status, followed by search of germ-line mutation in patients with MSI CRC. RESULTS: Synchronous colorectal cancers (50/881, 5.7%) were associated with stage IV microsatellite-stable (MSS) CRC (19/205, 9.3%, P=0.001) and with HNPCC (9/32, 28%, P<0.001). BRAF mutation (60/881, 6.8%) was associated with sporadic MSI CRC (37/62, 60%, P<0.001) but not with S-CRC (3/50, 6.0%, P=0.96). Synchronous colorectal cancer (HR 1.82; 95% CI 1.15-2.87; P=0.01), synchronous advanced adenoma (HR 1.81; 95% CI 1.27-2.58; P=0.001), and BRAF(c.1799T>A) mutation (HR 2.16; 95% CI 1.25-3.73; P=0.01) were stage-independent predictors of death from MSS CRC. Disease-specific survival of MSI CRC patients was not affected by S-CRC (HR 0.74; 95% CI 0.09-5.75; P=0.77). CONCLUSION: Microsatellite-stable CRCs have a worse prognosis if S-CRC or synchronous advanced adenoma are diagnosed. The occurrence and the enhanced aggressiveness of synchronous MSS advanced neoplasia are not associated with BRAF mutation.

Effect of ultrasound treatment on the water state in kiwifruit during osmotic dehydration.

The present work investigates how ultrasound pretreatment modulates the effects of osmotic dehydration (OD) on the water state and microstructure of kiwifruit. Kiwifruit slices (10mm thick) were subjected to ultrasonic waves in a water bath at a frequency of 35 kHz for 10, 20 and 30 min. OD process was then carried out by immersing the samples in 61.5% sucrose solution equilibrated at 25°C for a contact period of 0, 10, 20, 30, 60 and 120 min. The partition of water into the cellular tissue structures (vacuole, cytoplasm, extracellular spaces and cell wall) was investigated by Time Domain Nuclear Magnetic Resonance (TD-NMR). In parallel, the microstructure of kiwifruits slices was examined using a Scanning Electron Microscope. The results showed that US pretreatment performed for more than 10 min had a positive effect on the mass exchange caused by osmotic dehydration. A creation of microchannels and an increase of the average cross-section area of cells were observed when the samples were pretreated with US before OD. TD-NMR showed a slight redistribution of water through the substructures of the cells, as a function of the length of the US pretreatment applied.

Early expression of the fractalkine receptor CX3CR1 in pancreatic carcinogenesis.

BACKGROUND: In pancreatic ductal adenocarcinoma (PDAC), fractalkine receptor CX3CR1 contributes to perineural invasion (PNI). We investigated whether CX3CR1 expression occurs early in PDAC and correlates with tumour features other than PNI. METHODS: We studied CX3CR1 and CX3CL1 expression by immunohistochemistry in 104 human PDAC and coexisting Pancreatic Intraepithelial Neoplasia (PanIN), and in PdxCre/LSL-Kras(G12D) mouse model of PDAC. CX3CR1 expression in vitro was studied by a spheroid model, and in vivo by syngenic mouse graft of tumour cells. RESULTS: In total, 56 (53.9%) PDAC expressed CX3CR1, 70 (67.3%) CX3CL1, and 45 (43.3%) both. CX3CR1 expression was independently associated with tumour glandular differentiation (P=0.005) and PNI (P=0.01). Pancreatic Intraepithelial Neoplasias were more frequently CX3CR1+ (80.3%, P<0.001) and CX3CL1+ (86.8%, P=0.002) than matched cancers. The survival of PDAC patients was better in those with CX3CR1+ tumour (P=0.05). Mouse PanINs were also CX3CR1(+) and -CL1(+). In vitro, cytokines significantly increased CX3CL1 but not CX3CR1 expression. Differently, CX3CR1 was upregulated in tumour spheroids, and in vivo only in well-differentiated tumours. CONCLUSION: Tumour differentiation, rather than inflammatory signalling, modulates CX3CR1 expression in PanINs and PDAC. CX3CR1 expression pattern suggests its early involvement in PDAC progression, outlining a potential target for interfering with the PanIN transition to invasive cancer.

Van-Gogh-like 2 antagonises the canonical WNT pathway and is methylated in colorectal cancers.

BACKGROUND: Aberrant activation of the canonical WNT signaling is a feature of colorectal cancer (CRC). Van-Gogh-like 2 (VANGL2) belongs to the non-canonical WNT pathway whose activation inhibits canonical WNT signaling. In this study, we investigated the role of VANGL2 and its epigenetic regulation in CRC. METHODS: Van-Gogh-like 2 expression and promoter methylation after 5-aza-2'-deoxycytidine (5-aza) treatment were evaluated in CRC cells. DNA samples from 418 sporadic CRCs were tested for VANGL2 promoter methylation and microsatellite instability (MSI). Proliferation, colony formation and activation of the WNT pathway were tested in cells after VANGL2 overexpression. RESULTS: Van-Gogh-like 2 mRNA was significantly higher in 5-aza-treated RKO, LOVO and SW48, whereas no differences were found in SW480. Van-Gogh-like 2 was fully methylated in RKO, SW48, HCT116, DLD1 and Caco2; partially methylated in LOVO, LS174T and SW837; and unmethylated in SW480, SW620 and HT29. Higher expression of VANGL2 mRNA was found in the unmethylated cell lines. In CRC specimens (8.93% MSI), methylated VANGL2 was associated with MSI, higher grade, proximal colon location and BRAF mutation. Van-Gogh-like 2 overexpression in SW480 significantly decreased proliferation, colony formation and ß-catenin levels. CONCLUSION: Van-Gogh-like 2 is frequently methylated in MSI-CRCs with BRAF mutation and may act as a tumour suppressor gene, counteracting WNT/ß-catenin signaling.

The use of sodium bicarbonate for marination of broiler breast meat.

This study aimed to evaluate marination performances and the effect on meat quality traits of sodium bicarbonate, used alone or in combination with sodium chloride, when compared with sodium trypolyphosphate by using advanced analytical tools, including low-field nuclear magnetic resonance and differential scanning calorimetry. In total, 140 samples (cylindrical shape of 1 × 4 cm size) were obtained from a batch of 24-h postmortem broiler breast meat (Ross 708, females, 47 d old). Six of the groups were used for subsequent marination treatments, whereas the last group was kept as a nonmarinated control. Samples were subjected to vacuum tumbling in a special equipped laboratory rotary evaporator with a 12% (wt/wt) water:meat ratio using 6 marinade solutions: 7.7% (wt/wt) NaCl (S); 2.3% (wt/wt) Na(4)O(7)P(2) (P); 2.3% (wt/wt) NaHCO(3) (B); S and P; S and B; S, P, and B. Samples marinated with bicarbonate alone or in combination (B, SB, and SPB) significantly increased (P < 0.05) the meat pH by approximately 0.7 units compared with that of the control, whereas phosphate alone or in combination with salt increased (P < 0.05) the pH by 0.2 units. The combination containing all of the ingredients (SPB) produced the highest marinade performances; however, SB was able to guarantee a better marinade uptake and water retention ability with respect to that of SP. According to low-field nuclear magnetic resonance, the combined use of B and P with S determined a remarkable increase in proportion of entrapped water into the myofibrillar spaces, while the extramyofibrillar water fraction was not modified. Moreover, water gain following marination does not correspond to an increase in the freezable water amount, as detected by differential scanning calorimetry. In conclusion, B is a very promising marinating agent, and it can be exploited to develop processed poultry products with no added phosphates to match the request to avoid the nutritional drawbacks recently indicated with the use of phosphates.

Blood pressure lowering effect of lactotripeptides assumed as functional foods: a meta-analysis of current available clinical trials.

The oral assumption of lactotripeptides Valine-Proline-Proline (VPP) and Isoleucine-Proline-Proline (IPP) as nutraceuticals or functional foods is supposed to improve blood pressure (BP) control by angiotensin-converting enzyme-inhibition. However, data derived from clinical trials have reached conflicting conclusions. To perform a meta-analysis of placebo-controlled clinical trials evaluating the anti-hypertensive effect of lactotripeptides assumed as nutraceuticals or functional foods. Trials identified using a defined search strategy in PubMed were included in the meta-analysis, and their pooled effect was estimated with a random effects model, weighting for the inverse of the variance. Heterogeneity, publication bias, subgroup and meta-regression analyses were performed. A total of 18 trials have been identified, the clinical data of which have been clearly reported. Pooled effect of peptides was a reduction of -3.73 mm Hg (95% CI: -6.70, -1.76) for systolic blood pressure (SBP) and 1.97 mm Hg (95% CI: -3.85, -0.64) for diastolic blood pressure (DBP). The effect was more evident in Asian patients (SBP = -6.93 mm Hg (95% CI: -10.95, -2.94); DBP=-3.98 mm Hg(95% CI: -5.38, -2.44)) than in Caucasian ones (SBP=-1.17 mm Hg (95% CI: -2.82, 0.72); DBP = -0.52 mm Hg (95% CI: -1.39, 0.13)), and apparently not related to age, baseline BP values, dose of lactotripeptides assumed or length of the treatment. VPP and IPP lactotripeptides assumed as functional foods may significantly reduce SBP particularly in Asian subjects. The relevance of this findings in other ethnicities or associated with different dietary pattern should to be further investigated.

Influence of lifestyle measures on hypertriglyceridaemia.

Hypertriglyceridaemia is a common dyslipidaemia encountered in clinical practice. People with hypertriglyceridaemia are frequently obese, insulin-resistant, hypertensive or diabetic, all of which are risk factors for cardiovascular diseases. Hypertriglyceridaemia also contributes to metabolic syndrome, in which an atherogenic diet, sedentary lifestyle, overweight/obesity and genetic factors interact. A multi-factorial intervention for all risk factors is necessary, including weight reduction, dietary modification and increased physical exercise. This review focuses on the influence of diet, sedentary lifestyle and negative habits (such as excessive alcohol intake, smoking and drug addiction) on hypertriglyceridaemia as well as the effects of lifestyle change.

Differences and evolution of the methods for the assessment of microsatellite instability.

Microsatellite instability (MSI) originates from the systematic accumulation of uncorrected deletion/insertion in repetitive DNA tracts in cancer cells with a deficient mismatch repair system. Among colorectal cancers, the MSI signature identifies hereditary cases arising in patients with germline mutations in hMLH1, hMSH2, PMS2 and a fraction of those with hMSH6 mutations, as well as sporadic cancers with epigenetic hMLH1 promoter hypermethylation. Considering the specific pathogenesis, pathological features, natural history and response to 5-fluoro-uracil-based chemotherapy of the MSI cancers, confusion about the genetic markers for MSI recognition seems surprising. In this clinically relevant field, an agreement has not been reached concerning the use of di- or mononucleotide markers for MSI assessment. The Revised Bethesda Guidelines still recommend a panel of markers consisting of mono- and dinucleotides, despite being questioned whether it is congruous to continue to use dinucleotide markers for MSI identification. In any event, no single marker is accurate enough for MSI testing, and an awareness of their pros and cons is required for proper interpretation of results. In recent years, several papers have reported different prevalence of MSI in unrelated series, largely depending on the detection and classification method, suggesting that MSI test interpretation also requires the understanding of the phenomenon rather than simply the crude satisfaction of panel recommendations. Inaccuracies can otherwise lead to under- or overdiagnosis and inaccurate disease classification, which always have a negative impact on the clinical practice of medicine.