Endogenous skeletal muscle antioxidants

Skeletal muscle is susceptible to oxidative deterioration due to a combination of lipid oxidation catalysts and membrane lipid systems that are high in unsaturated fatty acids. To prevent or delay oxidation reactions, several endogenous antioxidant systems are found in muscle tissue. These include α‐tocopherol, histidine‐containing dipeptides, and antioxidant enzymes such as glutathione peroxidase, superoxide dismutase, and catalase. The contribution of α‐tocopherol to the oxidative stability of skeletal muscle is largely influenced by diet. Dietary supplementation of tocopherol has been shown to increase muscle α‐tocopherol concentrations and inhibit both lipid oxidation and color deterioration. Dietary selenium supplementation has also been shown to increase the oxidative stability of muscle presumably by increasing the activity of glutathione peroxidase. The oxidative stability of skeletal muscle is also influenced by the histidine‐containing dipeptides, carnosine and anserine. Whereas carnosine and anserine are affected by diet less than α‐tocopherol and glutathione peroxidase, their concentrations vary widely with species and muscle type. In pigs, beef, and turkey muscle, carnosine concentrations are greater than anserine, while the opposite is true in rabbit, salmon, and chicken muscle. Anserine and carnosine are found in greater concentrations in muscle high in white fibers, with chicken white muscle containing over fivefold more anserine and carnosine than red muscle. Anserine and carnosine are thought to inhibit lipid oxidation by a combination of free radical scavenging and metal chelation.     

An evaluation of a method to differentiate the species of origin of meats on the basis of the contents of anserine,balenine and carnosine in skeletal muscle

The high performance liquid chromatographic method proposed by Carnegie for determining the species of origin of meats in cooked products has been evaluated. The dipeptides anserine, balenine (ophidine) and carnosine have been extracted from the skeletal muscle of animals in New Zealand and compared with results obtained in Australia. The presence of significant quantities of balenine in red deer meat is reported for the first time. Limitations of the method are discussed.     

Identifying critical parameters for extraction of carnosine and anserine from chicken meat with high voltage pulsed electric fields and water

Carnosine (β-alanyl-L-histidine) and its methylated counterpart anserine (β-alanyl-1-methylhistidine) are important functional dipeptides found in various vertebrates' tissues. In this study, we identified the critical parameters of pulsed electric fields (PEF), coupled to mechanical pressing, followed by incubation in water up to 240 min for the extraction of these bioactive dipeptides from chicken meat. We show that PEF improves the kinetics of anserine and carnosine release to water by 7 to 53%, compared to the same water extraction process without PEF when the incubation time is below 120 min. A fractional factorial design showed that the incubation time in the water, after PEF pretreatment, had the most significant effect on dipeptides extraction. The maximum achieved total protein yield was 15.5 mg g_{fresh weight(FW)} ^-1 after 120 min incubation in water. The maximum carnosine extraction yield was 7.48 mg g_FW⁻¹ with a maximum anserine extraction yield of 3.05 mg g_FW⁻¹. The specific energy yield of extraction of protein, carnosine, and anserine was 488.16 mg J⁻¹, 39.43 mg J⁻¹, and 131.63 mg J⁻¹ respectively. The developed method is scalable and could be further explored to extract bioactive compounds from animal tissues.     

Use of histidine dipeptides to estimate the proportion of pig meat in processed meats

High performance liquid chromatography on a Partisil-10 SCX column was examined as an alternative to conventional ion exchange chromatography as a means of quantitating the histidine dipeptides, anserine, carnosine and balenine in processed pig meats. Because the ratio of these dipeptides in skeletal muscle of the pig is different from that in other species commonly used for meat, it is possible to estimate the proportion of pig meat in processed meats. Several Australian pork products were found to contain a low proportion of lean pig meat.     

Influence of dietary β-alanine and histidine on the oxidative stability of pork

Carnosine (β-alanine-L-histidine) and anserine (β-alanine-1-methyl histidine) are endogenous antioxidants found in skeletal muscle. The objective of this research was to determine if supplementation of swine diets with histidine (histidine; 0.40%) and/or β-alanine (β-alanine; 0.225%) was an effective method to increase carnosine and anserine concentrations and the oxidative stability of Longissimus dorsi (LD) and Vastus intermedius (VI) muscles. Dietary treatments had no effect on carnosine and anserine concentrations in LD; however, histidine + β-alanine supplementation increased carnosine and anserine in VI muscle compared to β-alanine supplementation. Dietary supplementation had no effect on the formation of thiobarbituric acid reactive (TBARS) or lipid peroxides in cooked VI and LD. In salted VI and LD muscle, differences in TBARS and peroxides were observed; however, these differences did not consistently correlate with differences in anserine and carnosine concentrations. Therefore, the results of this research suggest that supplementation of swine diets with β-alanine and/or histidine is not an efficient method to increase the oxidative stability of pork.     

Bioactivities of chicken essence

The special flavor and health effects of chicken essence are being widely accepted by people. Scientific researches are revealing its truth as a tonic food in traditional health preservation. Chicken essence has been found to possess many bioactivities including relief of stress and fatigue, amelioration of anxiety, promotion of metabolisms and post-partum lactation, improvement on hyperglycemia and hypertension, enhancement of immune, and so on. These activities of chicken essence are suggested to be related with its active components, including proteins, dipeptides (such as carnosine and anserine), polypeptides, minerals, trace elements, and multiple amino acids, and so on. Underlying mechanisms responsible for the bioactivities of chicken essence are mainly related with anti-stress, anti-oxidant, and neural regulation effects. However, the mechanisms are complicated and may be mediated via the combined actions of many active components, more than the action of 1 or 2 components alone.     

EFFECT OF CARNOSINE, ANSERINE AND OTHER ENDOGENOUS SKELETAL PEPTIDES ON THE ACTIVITY OF PORCINE MUSCLE ALANYL AND ARGINYL AMINOPEPTIDASES

The effect of endogenous dipeptides (carnosine and anserine) and peptidic fractions from meat extracts on aminopeptidase activity, alanyl and arginyl aminopeptidases purified from porcine skeletal muscle, was studied. Carnosine inhibited the activity of porcine arginyl aminopeptidase (RAP); the inhibition being stronger with the purified form of the enzyme than in the muscle extract. The RAP inhibition was competitive. One of the peptidic fractions isolated from meat extracts inhibited RAP activity But the degree of inhibition depended on the extent of purification. The major aminopeptidase present in muscle, alanyl aminopeptidase (AAP), was neither inhibited by natural dipeptides nor arty of the peptidic fractions. The study of meat inhibitory peptides provides a better understanding of the action of proteolytic enzymes during meat processing.     

FREE AMINO ACIDS AND DIPEPTIDES IN DRY-CURED HAM

Six types of dry-cured hams were chemically profiled for proximate composition and low molecular weight nonprotein nitrogen (NPN) fractions including free amino acids (FAA) and two dipeptides (carnosine and anserine). Results indicate that FAA were present in the six classes at different (mg/100g protein) concentrations, with Iberian and Serrano containing greater (P<0.05) amounts of most FAA. However, when analyzed as percentage (mg/100g total FAAs) values, FAA contents of Iberian and Serrano hams exhibited lower (P<0.05) tyrosine and greater (P<0.05) aspartic acid and threonine values. Within each class, large variation coefficients (>10%) for nonprotein nitrogen paralleled inconsistent values of salt content, to which NPN and FAAs were negatively correlated (P<0.01). NPN was also negatively affected by moisture in the dried muscle, indicating limited saltiness and greater muscle shrinkage as two major determinants in developing of nonprotein fractions, hence of free amino acids.     

Effect of cooking method on carnosine and its homologues, pentosidine and thiobarbituric acid-reactive substance contents in beef and turkey meat

Commercial samples of beef and turkey meat were prepared by commonly used cooking methods with standard cooking times: (1) broiled at 200 °C for 10 min, (2) broiled at a medium temperature (140 °C) for 10 min, (3) cooked by microwave (MW) for 3 min and then grilled (MW/grill) for 7 min, (4) cooked in a domestic microwave oven for 10 min, and (5) boiled in water for 10 min. The raw and cooked meats were then analysed to determine the carnosine, anserine, homocarnosine, pentosidine, and thiobarbituric acid-reactive substance (TBARS) contents. It was observed that boiling beef caused a loss of approximately 50% of the carnosine, probably because of the high water solubility of carnosine and its homologues; cooking by microwave caused a medium loss of the anti-oxidants of approximately 20%; cooking by MW/grill led to a reduction in carnosine of approximately 10%. As far as the anserine and homocarnosine contents were concerned, a greater loss was observed for the boiling method (approximately 70%) while, for the other cooking methods, the value ranged from 30% to 70%. The data oscillate more for the turkey meat: the minimum carnosine decrease was observed in the cases of MW/grill and broiling at high temperature (25%). Analogously, the anserine and homocarnosine contents decreased slightly in the case of MW/grill and broiling at a high temperature (2-7%) and by 10-30% in the other cases. No analysed meat sample showed any traces of pentosidine above the instrumental determination limits. The cooked beef showed an increased TBARS value compared to the raw meat, and the highest values were found when the beef was broiled at a high temperature, cooked by microwave or boiled in water. The TBARS value of the turkey meat decreased for all the cooking methods in comparison to the TBARS value of the fresh meat.     

Electrochemical profiling using copper nanoparticle-plated electrode for identification of ostrich meat and evaluation of meat grades

Electrochemical (EC) profiling employing copper nanoparticle-plated screen-printed electrode (Cuⁿ-SPE) exhibited specific selectivity and sensitivity to peptides that could efficiently differentiate ostrich meat from pork, beef and chicken, and to evaluate grades and freshness of ostrich meat. The four meats were differentiated in 5 min by characteristic chromatographic profiles consisting only four major peaks. Peak identification by LC/MS/MS suggested that while lysine and glutamine were shown as common components, anserine were the avian-specific and carnosine was the ostrich-missing peptide in the EC profile. Statistical analysis (ROC curve) demonstrated that peak ratios could be used to evaluate ostrich meat grades with high sensitivity (up to 95%) and specificity (up to 100%). The effects of storage temperature and time were studied for potential use of Cuⁿ-SPE to evaluate meat freshness. This HPLC-EC method appeared to be superior to UV detection in terms of profile simplicity and devoid of derivatisation process or complex sample extraction procedures, rendering it a suitable method for routine rapid differentiation of ostrich meat from common meat species with the ability to simultaneously differentiate ostrich meat grades.     

Nutritional pork meat compounds as affected by ham dry-curing

This work is focused on the determination of compounds of nutritional interest that are naturally present in pork meat and how they are affected during the processing of dry-cured ham. Such compounds are creatine, creatinine, coenzyme Q₁₀, glutathione, carnosine, anserine, carnitine, taurine, cystine, cysteine and the essential amino acids. Their antioxidant and antyhipertensive functions were evaluated. Of all the assayed substances, only glutathione decreased totally during processing. Carnosine, creatinine, anserine and glutathione showed antioxidant, while cysteine, glutathione and carnosine showed antyhipertensive activity. So, dry-cured ham constitutes an excellent source of essential amino acids (all essential amino acids exhibited a large increase during processing) and other nutritionally interesting compounds such as cystine, cysteine, carnosine, anserine, taurine, carnitine and coenzyme Q₁₀.     

Investigations on the origin of two radiation-induced compounds in irradiated meat

Summary After irradiation with 0.5 Mrad or higher doses, a radiation-induced ninhydrin-positive compound was found in the muscle tissue of warm-blooded animals. In carp muscle another compound called X was formed. The origin of these two compounds was investigated. Irradiation of various amino acids and related compounds showed that substance Y is formed from the dipeptides carnosine and anserine, which are present in muscle tissue of warm-blooded animals in unusually high concentrations. In carp muscle containing relatively high levels of histidine, but only low concentrations of anserine as the only dipeptide, histidine furnished compound X. This compound was found to be relatively stable in acid medium. After acid hydrolysis of irradiated carnosine solutions and of extracts of irradiated muscle tissue from warm-blooded animals, compound Y was no longer observed, but another compound appeared that was similar to X with regard to colour and migration properties during high-voltage electrophoresis and column chromatography.

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Zur Herkunft zweier bei der Bestrahlung von Fleisch gefundener Bestrahlungsprodukte

Zusammenfassung Nach der Bestrahlung mit 0,5 Mrad oder höheren Dosen war im Warmblütermuskel ein mit Y und im Karpfenmuskel ein anderes mit X bezeichnetes ninhydrinpositives Bestrahlungsprodukt gefunden worden. Die Herkunft dieser beiden Komponenten wurde untersucht. Die Bestrahlung von verschiedenen Aminosäuren und verwandten Verbindungen ergab, daß die Substanz Y aus den im Warmblütermuskel in unverhältnismäßig hohen Konzentrationen vorhandenen Dipeptiden Carnosin und Anserin entsteht. Im Karpfenmuskel, in dem relativ große Gehalte an Histidin, aber von den Dipeptiden nur Anserin in geringen Konzentrationen gefunden wurde, lieferte Histidin die unter sauren Bedingungen relativ beständige Komponente X. Bei der sauren Hydrolyse von bestrahlten Carnosinlösungen und von Extrakten der bestrahlten Warmblütermuskulatur verschwand das Bestrahlungsprodukt Y. Dafür trat eine Komponente auf, die in Färbung und in den Laufeigenschaften bei der Hochspannungselektrophorese und Säulenchromatographie mit X identisch war.

     

Pork meat quality affects peptide and amino acid profiles during the ageing process

Twenty pork carcasses were classified in different pork meat qualities: red, firm and non-exudative (RFN), pale, soft and exudative (PSE), red, soft and exudative (RSE) and dark, firm and dry (DFD) meat. The content of peptides and free amino acids during the ageing process was analysed and compared within quality classes. Four peptide fractions were isolated through cation-exchange and reverse-phase chromatography. The main significant differences among qualities were obtained for peptide fractions 3 and 4. Peptide fraction 3 at 4 days and peptide fraction 4 at 2 h postmortem were higher in the ideal pork quality (RFN) than in the other quality classes. The ageing of pork meats produced a general increase in all free amino acid concentrations for the studied quality classes except for Gln, β-Ala, Taurine and Orn and the dipeptides carnosine and anserine. The DFD class showed higher increases in Lys, Ala and Met probably due to the activation of neutral aminopeptidases.     

The Inhibition of Advanced Glycation End Products by Carnosine and Other Natural Dipeptides to Reduce Diabetic and Age‐Related Complications

As people age they are at a greater risk for many disorders including cardiovascular, renal, and neurodegenerative diseases, and these conditions are exacerbated by diabetes. An important cause of the maladies associated with both age and diabetes is the formation of advanced glycation end products (AGEs). AGE formation is initiated by glycation reactions between reducing sugars and free amine groups. A cascade of other reactions follows, leading to alterations in membrane function and damage to the proteome, such as protein crosslinking. Compounds that prevent these reactions are currently being researched, but peptides hold great potential as they tend to lack toxicity, are absorbed intact, are easily produced, and are cheaper than other options. Of the peptides researched, carnosine is the most promising. Research suggests that carnosine is absorbed into the plasma unaltered and intact. Carnosine has been shown to prevent AGE formations through reduction of blood glucose, prevention of early glycation, and even reversing previously formed AGEs. Other promising peptides and amino acids include β‐alanine, L‐histidine, homocarnosine, anserine, and glutathione. If bioactive peptides and amino acids can minimize the formation of AGEs, foods containing these peptides could be used to improve health.     

Free amino acids and dipeptides in porcine muscles: differences between `red' and `white' muscles

Three porcine muscles (m. longissimus dorsi, masseter and trapezius), chosen to represent the three main metabolic types, from 18 carcasses had their free amino acids and dipeptides quantified by reverse-phase high performance liquid chromatography (HPLC) in aqueous extracts derivatized with phenyl isothiocyanate. Of the 25 measured compounds, four amino acids and the dipeptide carnosine were closely related to the metabolic type of the three muscles. Masseter, a red oxidative muscle, had the highest contents of aspartic acid, glutamine and taurine. Longissimus dorsi, a white glycolytic muscle was characterised by the highest contents of β-alanine and carnosine. Trapezius, an intermediate muscle, had intermediate contents. These results show that free amino acid and dipeptide contents could partly explain differences in taste of muscles from the same species.     

The detection of chicken meat in meat products by means of the anserine/carnosine ratio

A distinctive difference was found between the ratio of the anserine and carnosine contents (a/c ratio) in beef or pork and of that in chicken/meat. The a/c ratio for beef varies between 0.06–0.2 and for pork between 0.02–0.1 but for chicken meat can reach values as high as 2.2–5.5.The high a/c ratio for chicken meat proved to be sufficient to detect this ingredient at a 5% level in both cooked pork products and 1:1 beef-pork mixtures, this being independent of the heat treatment.     

Monoclonal antibodies against troponin I for the detection of rendered muscle tissues in animal feedstuffs

Regulatory controls to prevent the spread of BSE have prohibited the use of certain animal proteins in feed in several countries. Accurate analytical methods for detecting prohibited material in feedstuffs are needed to ensure compliance with the new regulations. Six IgG class monoclonal antibodies (MAbs) against troponin I (TnI), a thermostable marker protein, have been developed for the detection and differentiation of rendered muscle tissue in animal feed. MAbs 1F9, 2G3 and 7F7 reacted to TnI of all species, including mammalian, poultry and fish, while MAbs 7A12 and 8A12 recognized only mammalian TnI (porcine, bovine, ovine, equine, and deer). MAb 2A8 was able to differentiate TnI of ruminant origin (bovine, ovine and deer) from other species. Three indirect enzyme-linked immunosorbent assays (ELISAs) employing these MAbs were developed for the determination of animal muscle, mammalian muscle or ruminant muscle in animal feeds.     

Genetically Modified Feeds in Poultry Diet: Safety,Performance, and Product Quality

Concerns have been expressed regarding the safety of using biotechnology derived feeds in diets of livestock animals and in regard to human consumption of products from species fed transgenic crops. As a consequence, a large number of poultry nutrition studies have been conducted to evaluate the wholesomeness of transgenic crops by examining performances of animals during growth or egg laying. Studies also evaluated whether foreign DNA and proteins could be detected in meat, egg, and tissue samples from broiler chickens and laying hens fed diets containing transgenic feeds. In all studies, the conclusions were in agreement that the transgenic crops provided comparable performance, carcass and egg yields, and meat and egg composition, when compared with conventional grains. Moreover, it was demonstrated that transgenic proteins and DNA present in livestock feeds are not detectable in food products derived from these animals, using the most sensitive detection methods available, confirming that they are rapidly degraded by normal digestive processes. The lack of significant differences were a result of the similarity in nutrient composition of the genetically modified feeds and lack of differences in intake and digestibility, while there were no evidences that the differences reported for performance response variables and carcass measurements between treatment groups were attributable to the presence of the transgenic gene and protein in the biotechnology derived plants. Results demonstrated that genetically modified feeds are substantially equivalent and they result as safe as existing conventional feeds.     

Hypolipidemic and Anti-Obesity Effect of Anserine on Mice Orally Administered with High-Fat Diet via Regulating SREBP-1, NLRP3, and UCP-1

To investigate the efficacy of anserine on antiobesity, C57BL/6 mice are orally administered with a high-fat diet (HFD) and different doses of anserine (60, 120, and 240 mg/kg/day) for 16 weeks. Body weight, lipid, and epididymal fat content in mice are measured, and their liver damage is observed. The results display that the body weight, epididymal fat content, and low-density lipoprotein cholesterol (LDL-C) content in anserine groups are decreased by 4.36–18.71%, 7.57–35.12%, and 24.32–44.40%, respectively. To further investigate the antiobesity mechanism of anserine, the expression of SREBP-1, NLRP3, NF-κB p65 (p65), and p-NF-κB p65 (p-p65) proteins in the liver and peroxisome proliferator-activated receptor gamma coactivator 1α (PGC1-α) and UCP-1 proteins in brown adipose tissue (BAT) is analyzed by Western blot. Results show that anserine can significantly decrease the expression of the NLRP3, p65, p-p65, and the SREBP-1 proteins and increase the expression of the PGC1-α and UCP-1 proteins. This study demonstrates that anserine lowered blood lipids and prevented obesity; its antiobesity mechanism may be related to the activation of brown fat by inflammation.     

In vitro digestibility based on fish crude enzyme extract for prediction of feed quality in growth trials

Biochemical structure of protein (reactive SH content, content ratio of SH/S? S and concentration of D ‐Asp as % of total (D + L )‐Asp) indicating digestibility of dietary protein was changed under different processing conditions. Based on fish crude enzyme extract, in vitro digestibility of different fish materials processed under different conditions correlated positively with reactive SH content and content ratio of SH/S? S and negatively with D ‐Asp concentration. In vitro digestion of different experimental feeds, based on Atlantic salmon crude enzyme extracts, was studied in association with growth trials in order to investigate its value as a criterion for industrial strategy in predicting feed quality. Crude enzymes were extracted from the pyloric caeca before feeding. Significant differences in in vitro digestibility between the experimental feeds were observed whereby there would be differences in feed conversion efficiency within 3 months of feeding. There were associations between the in vitro digestibility and other parameters for dietary quality, such as mink digestibility and the biochemical structure parameters of the dietary protein due to different processing conditions. Crude enzyme extracts from rainbow trout and European seabass were also used for in vitro digestibility study of different experimental feeds by standardising trypsin activity to that of Atlantic salmon crude enzyme extract. The results indicated that different fish species have different digestion ability to the same feed types, and the effective time for feed utilisation and growth is dependent on fish sensitivity and the extent of difference in digestibility between the feeds consumed as observed in the Atlantic salmon trials. For the species investigated, sensitivity ranking of the enzymes to feed quality under the condition studied was Atlantic salmon > rainbow trout > European seabass. The results indicated that in vitro digestibility study of experimental feeds using pyloric caecal crude enzyme extract from a specific species at an age of interest could be a practical, quick and reliable method for testing feed quality in growth trials. By standardising the crude enzyme extract with regards to trypsin activity, the in vitro digestibility values could be comparable not only within the same species but also between different species. © 2002 Society of Chemical Industry