Validation of a 1H-NMR Spectroscopy Quantitative Method to Quantify Trimethylamine Content and K -Index Value in Different Species of Fish

The chemical analysis that is frequently employed for the evaluation of the freshness of fish includes (i) the quantification of trimethylamine (TMA) and (ii) the estimation of the K-Index, based on the ratio between the concentrations of adenosine triphosphate (ATP) and its breakdown products. TMA is quantified using a colorimetric reference method (AOAC), while the K-Index is usually determined by HPLC. The present work proposes a method for the above freshness biomarkers based on HR H-1-NMR as an alternative method able to assess both indexes simultaneously on aqueous fish extracts. To validate the proposed H-1-NMR method, a large set of validation checks has been addressed, such as accuracy, precision, specificity, limits of detection, linearity, and range of linearity and quantification according to EuroChem guidelines. The results show that the methodology satisfies all the validation requirements at the same level as the most frequently used methods, with the advantage of being faster and more repeatable, avoiding the use of solvents, such as toluene and formaldehyde, or dangerous reagents such as picric acid

First insights into the urinary metabolome of captive giraffes by proton nuclear magnetic resonance spectroscopy

The urine from 35 giraffes was studied by untargeted1H-NMR, with the purpose of obtaining, for the first time, a fingerprint of its metabolome. The metabolome, as downstream of the transcriptome and proteome, has been considered as the most representative approach to monitor the relationships between animal physiological features and environment. Thirty-nine molecules were unambiguously quantified, able to give information about diet, proteins digestion, energy generation, and gut-microbial co-metabolism. The samples collected allowed study of the effects of age and sex on the giraffe urinary metabolome. In addition, preliminary information about how sampling procedure and pregnancy could affect a giraffe\u2019s urinary metabolome was obtained. Such work could trigger the setting up of methods to non-invasively study the health status of giraffes, which is utterly needed, considering that anesthetic-related complications make their immobilization a very risky practice

Lack of mutation at codon 531 of SRC in advanced colorectal cancers from Italian patients

A truncating mutation (C to T transition) at codon 531 of the human protooncogene c-src, possibly accounting for the activation of c-src tyrosine kinase, has been recently identified in a subset of advanced colorectal cancer from North-American patients. However, two subsequent studies have failed to confirm the occurrence of SRC 531 mutation in colorectal cancers from North-European and Asiatic patients, raising the hypothesis that the genetic activation of src in colon cancer might be restricted to patients belonging to specific ethnic groups. We investigated a large series of colorectal cancers from Italian patients (155 cases) with a high prevalence of liver metastasis (43%). Using a PCR-RFLP assay, the occurrence of a SRC 531 mutation was ruled out in all the investigated specimens of primary tumours and/or metastases. Our results demonstrate that SRC Gln531AMB plays no role in the development or in the progression of colorectal cancer among Italian patients. © 2001 Cancer Research Campaign http://www.bjcancer.co

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

ABSTRACT 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 (1 H-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 1 H-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 1 H-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

An Untargeted Metabolomics Investigation of Jiulong Yak (Bos grunniens) Meat by 1H-NMR

Yak represents the main meat source for Tibetan people. This work aimed to investigate the metabolome of raw meat from Jiulong yaks, focusing on specimens farmed and harvested locally through traditional procedures. Untargeted nuclear magnetic resonance spectroscopy (1H-NMR) was selected as the analytical platform. Samples from longissimus thoracis, trapezius, triceps brachii and biceps femoris muscles, with different prevalences of red and white fibers, were selected. Among the fifty-three metabolites quantified in each of them, carnitine, carnosine, creatine and taurine are known for their bioactive properties. Twelve molecules were found to be differently concentrated in relation to muscle type. Longissimus thoracis, compared to biceps femoris, had higher concentrations of carnosine and formate and lower concentrations of mannose, inosine, threonine, IMP, alanine, valine, isoleucine, tyrosine, phenylalanine and leucine. A metabolic pathway analysis suggested that the main pathways differing among the muscles were connected to the turnover of amino acids. These results contribute to a deeper understanding of yak raw meat metabolism and muscle type differences, which can be used as an initial reference for the meat industry to set up muscle-specific investigations. The possibility of simultaneously quantifying several bioactive compounds suggests that these investigations could revolve around meat's nutritional value

PEG reimplantation after Buried Bumper Syndrome: a case report

Percutaneous endoscopic gastrostomy (PEG) is the method of choice to provide long-term enteral nutrition for patients with impossibility to be fed orally. Although it is considered a routine and safe procedure, potential complications exist, which are generally classified into three major categories: endoscopic technical difficulties, PEG procedure-related complications and late complications associated with PEG tube use, such as buried bumper syndrome (BBS). BBS is a potentially life-threatening complication, occurring in 0.3% to 2.5% of cases. Additional complications related to BBS may present, such as wound infection, peritonitis, and necrotizing fasciitis. Once resolved the acute complication, an adequate feeding method should be prompted for the patient, among whom PEG remains of choice. After tissue inflammation, fibrosis may prevent a standard endoscopic procedure for the new implantation, therefore endoscopists should modulate procedures to obtain successful and safe results. A combined surgical-and endoscopic strategy could resolve implantation difficulties ensuring a safe and simple procedure. We present here a case of BBS complicated with abdominal wall cellulitis in a paraplegic 35-year-old-man who was admitted to our hospital. (www.actabiomedica.it)

Unusual sub-genus associations of fecal Prevotella and Bacteroides with specific dietary patterns

Background: Diet has a recognized effect in shaping gut microbiota. Many studies link an increase in Prevotella to high-fibre diet, while Bacteroides abundance is usually associated with the consumption of animal fat and protein-rich diets. Nevertheless, closely related species and strains may harbour different genetic pools; therefore, further studies should aim to understand whether species of the same genus are consistently linked to dietary patterns or equally responsive to diet variations. Here, we used oligotyping of 16S rRNA gene sequencing data to exploit the diversity within Prevotella and Bacteroides genera in faecal samples of omnivore and non-omnivore subjects from a previously studied cohort. Results: A great heterogeneity was found in oligotype composition. Nevertheless, different oligotypes within the same genus showed distinctive correlation patterns with dietary components and metabolome. We found that some Prevotella oligotypes are significantly associated with the plant-based diet but some are associated with animal-based nutrients, and the same applies to Bacteroides. Therefore, an indiscriminate association of Bacteroidetes genera with specific dietary patterns may lead to an oversimplified vision that does not take into account sub-genus diversity and the different possible responses to dietary components. Conclusions: We demonstrated that Prevotella and Bacteroides oligotypes show distinctive correlation patterns with dietary components and metabolome. These results substantiate a current oversimplification of diet-dependent microbe-host associations and highlighted that sub-genus differences must be taken into account when planning gut microbiota modulation for health benefits

Radiogenomics in clear cell renal cell carcinoma: correlations between advanced CT imaging (texture analysis) and microRNAs expression

Purpose: A relevant challenge for the improvement of clear cell renal cell carcinoma management could derive from the identification of novel molecular biomarkers that could greatly improve the diagnosis, prognosis, and treatment choice of these neoplasms. In this study, we investigate whether quantitative parameters obtained from computed tomography texture analysis may correlate with the expression of selected oncogenic microRNAs. Methods: In a retrospective single-center study, multiphasic computed tomography examination (with arterial, portal, and urographic phases) was performed on 20 patients with clear cell renal cell carcinoma and computed tomography texture analysis parameters such as entropy, kurtosis, skewness, mean, and standard deviation of pixel distribution were measured using multiple filter settings. These quantitative data were correlated with the expression of selected microRNAs (miR-21-5p, miR-210-3p, miR-185-5p, miR-221-3p, miR-145-5p). Both the evaluations (microRNAs and computed tomography texture analysis) were performed on matched tumor and normal corticomedullar tissues of the same patients cohort. Results: In this pilot study, we evidenced that computed tomography texture analysis has robust parameters (eg, entropy, mean, standard deviation) to distinguish normal from pathological tissues. Moreover, a higher coefficient of determination between entropy and miR-21-5p expression was evidenced in tumor versus normal tissue. Interestingly, entropy and miR-21-5p show promising correlation in clear cell renal cell carcinoma opening to a radiogenomic strategy to improve clear cell renal cell carcinoma management. Conclusion: In this pilot study, a promising correlation between microRNAs and computed tomography texture analysis has been found in clear cell renal cell carcinoma. A clear cell renal cell carcinoma can benefit from noninvasive evaluation of texture parameters in adjunction to biopsy results. In particular, a promising correlation between entropy and miR-21-5p was found