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1.
Reprod Biol ; 23(2): 100766, 2023 Jun.
Article in English | MEDLINE | ID: mdl-37084542

ABSTRACT

Bisphenol A (BPA) exposure may impair gonadal steroidogenesis, although the underlying mechanism is not well known. Hereby, we assessed BPA action on human primary granulosa (hGC) and mouse Leydig cells (BLTK-1) proliferation, cytotoxicity, hormone secretion, and steroidogenic enzyme/receptor gene profile. hGC and BLTK-1 cells were stimulated with increasing concentrations of BPA (10-12 M to 10-4 M for cell proliferation assay, 10-8 M to 10-4 M for LDH-cytotoxicity assay, and 10-9 M to 10-5 M for hormone secretion and genes expression analysis). BPA at low concentrations (pM - nM) did not affect cell proliferation in either cell type, although was toxic at higher (µM) concentrations. BPA stimulation at low nM concentrations decreased the production of estradiol (E2) and testosterone (T) in BLTK-1, E2, and progesterone in hGCs. BPA down-regulated Star, Cyp11a1, and Hsd17b3, but up-regulated Cyp19a1, Esr1, Esr2, and Gpr30 expression in BLTK-1 cells. In hGC, BPA down-regulated STAR, CYP19A1, PGRMC1, and PAQR7 but up-regulated ESR2 expression. Estrogen receptor degrader fulvestrant (FULV) attenuated BPA inhibition of hormone production in both cell lines. FULV also blocked the BPA-induced Gpr30 up-regulation in BLTK-1 cells, whereas in hGC, failed to reverse the down-regulation of PGRMC1, STAR, and CYP19A1. Our findings provide novel mechanistic insights into environmentally-relevant doses of BPA action through both nuclear estrogen receptor-dependent and independent mechanisms affecting cultured granulosa and Leydig cell steroidogenesis.


Subject(s)
Phenols , Receptors, Estrogen , Male , Mice , Animals , Humans , Phenols/toxicity , Progesterone/metabolism , Benzhydryl Compounds/toxicity , Membrane Proteins , Receptors, Progesterone
2.
Animals (Basel) ; 11(4)2021 Mar 25.
Article in English | MEDLINE | ID: mdl-33805968

ABSTRACT

The use of intensive selection procedure in modern broiler chicken lines has led to the development of several skeletal disorders in broiler chickens. Therefore, current research is focused on methods to improve the bone quality in birds. In ovo technology, using nanoparticles with a high specificity to bones, is a potential approach. The present study aimed to evaluate the effect of in ovo inoculation (IOI) of calcium carbonate nanoparticles (CCN) on chicken embryo development, health status, bone characteristics, and on broiler production results and bone quality. After assessing in vitro cell viability, the IOI procedure was performed with an injection of 500 µg/mL CCN. The control group was not inoculated with CCN. Hatchability, weight, and selected bone and serum parameters were measured in embryos. Part of hatchlings were reared under standard conditions until 42 days, and production results, meat quality, and bone quality of broilers were determined. CCN did not show cytotoxicity to cells and chicken embryo and positively influenced bone parameters of the embryos and of broilers later (calcification) without negatively affecting the production results. Thus, the IOI of CCN could modify the molecular responses at the stage of embryogenesis, resulting in better mineralization, and could provide a sustained effect, thereby improving bone quality in adult birds.

3.
J Sci Food Agric ; 101(7): 2634-2640, 2021 May.
Article in English | MEDLINE | ID: mdl-33159318

ABSTRACT

Biogenic amines (BA) are organic compounds commonly found in food, plants and animals, as well as microorganisms that are attributed with the production of BAs. They are formed as an effect of a chemical process: the decarboxylation of amino acids. Factors determining the formation of BAs include the availability of free amino acids and the presence of microorganisms that show activity with respect to carrying out the decarboxylation process. On the one hand, BAs are compounds that are crucial for maintaining cell viability, as well as the proper course of the organism's metabolic processes, such as protein synthesis, hormone synthesis and DNA replication. On the other hand, despite their positive effects on the functioning of the organism, an excessive content of BAs proves to be toxic (diarrhea, food poisoning, vomiting, sweating or tachycardia). Moreover, they can accelerate carcinogenesis. Amines are a natural component of plant and animal raw materials. As a result of the proven negative effects of amines on living organisms, the reduction of these compounds should be the subject of scientific research. The present review aims to synthesize and summarize the information currently available on BAs, as well as discuss the interpretation of the results. © 2020 Society of Chemical Industry.


Subject(s)
Biogenic Amines/analysis , Biogenic Amines/toxicity , Animals , Biogenic Amines/metabolism , Food Contamination/analysis , Foodborne Diseases/etiology , Foodborne Diseases/metabolism , Humans
4.
Animals (Basel) ; 10(10)2020 Oct 06.
Article in English | MEDLINE | ID: mdl-33036305

ABSTRACT

Meat from commercial breed cattle are very often used to crossbreed with dairy breeds. The effect of heterosis is most evident when crossbreeds are genetically different from each other. Therefore, the aim of the study was to determine the influence of breed types on the nutritional and pro-health quality of beef. The experiment was conducted on 62 bulls from three breeds: Limousin, Polish Holstein-Friesian, and Polish Holstein-Friesian (PHF) × Limousin. During the fattening period, the animals were fed ad libitum using the same diet. Bulls were slaughtered at 21-23 months of age. The meat of PHF × Limousin hybrids was characterized by the lowest level of SFA and the highest content of n-3 PUFA fatty acids, carnosine, and α-tocopherol compared to the values obtained for the Polish Holstein-Friesian and Limousin breeds. In the case of PHF × Limousin hybrids, there was a 6% increase in n-3 PUFA, 21% in carnosine, and 66% in α-tocopherol compared to the Polish Holstein-Friesian breed. Commodity crossbreeding significantly improved the quality of beef analyzed in this study, resulting in similar or even better results than purebred cattle. This meant that beef from the hybrids with PHF was of the best nutritional and health-promoting quality.

5.
Arch Anim Nutr ; 74(5): 343-361, 2020 Oct.
Article in English | MEDLINE | ID: mdl-32940083

ABSTRACT

Intensive selection in modern lines of fast-growing chickens has caused several skeletal disorders. Therefore, current research is focused on methods to improve the bones of birds. A new potential solution is in ovo technology using nanoparticles with a high specificity for the bone tissue. Thus, the objective of the present study was to evaluate the effect of in ovo application of hydroxyapatite nanoparticles (HA-NP) in different concentrations (50, 100 and 500 µg/ml colloids) on chicken embryo development, with a particular focus on the oxidative status and bone characteristics of the embryo. The results showed that in ovo treatment with HA-NP did not negatively affect hatchability and body weight. However, bone weight was reduced in 500 µg/ml group. The concentrations of calcium, phosphorus and crude ash were not affected. The modulatory effect of HA-NP was observed on the basis of antioxidative markers - superoxide dismutase, total antioxidant status, malondialdehyde in serum and selected tissues. Glutathione concentration in serum suggested higher metabolic stress. Among bone turnover markers, the concentration of osteocalcin was found to be significantly affected by HA-NP injection. Thus, the in ovo application of HA-NP could modify the molecular responses at the stage of embryogenesis but these changes were not reflected in embryo growth and even slowed down bone development. Nevertheless, the question for the follow-up research is whether in ovo administration of HA-NP would affect the antioxidative status and bone turnover resulting in improved bone conditions and body gain in post hatch chickens.


Subject(s)
Calcification, Physiologic/drug effects , Chick Embryo/drug effects , Durapatite/metabolism , Nanoparticles/metabolism , Ovum/drug effects , Oxidative Stress/drug effects , Animal Nutritional Physiological Phenomena , Animals , Chick Embryo/growth & development , Dose-Response Relationship, Drug , Durapatite/administration & dosage , Injections/veterinary , Nanoparticles/administration & dosage , Random Allocation
6.
Animals (Basel) ; 9(12)2019 Dec 02.
Article in English | MEDLINE | ID: mdl-31810335

ABSTRACT

Colostrum as a secretion of the mammary gland is produced and accumulated in the final stage of pregnancy and in the first days after calving. It is designed to provide the calf with the necessary nutrients and biologically active ingredients. One of the most difficult periods in the life of animals is their rearing, and the most sensitive are the first days after birth. This is the time when most falls occur, and they are caused by mortality and morbidity, even at the level of 30%. Such losses affect the performance and profitability of animal production (the percentage of animals intended for reproduction or fattening is reduced and the intensity of selection in the herd is also reduced). Both diseases and mortality are the cause of serious economic, production, and breeding losses, which are the result of weak immune mechanisms. The adaptability of calves to the environment is determined by their immune status. Colostrum has a regulating function and stimulates the young organism to grow, and it has properties that support the functioning of systems: Endocrine and immunological. For colostrum to fulfil its role, it must be administered immediately after birth, because the immunoglobulins it contains are absorbed during the first 16-27 h after the birth of the calf, preferably within 2-4 h of age. Blood from calves that have been properly calved should have an antibody concentration of 15g/L (24-48 h of age). Therefore, immunoglobulins are the most important factor affecting infectious immunity; an adequate concentration of immunoglobulins in calves' blood is related to their survival and health. It is the intent of this review to synthesize and summarize the information currently available on colostrum, as well as to discuss the interpretation of the results.

7.
Womens Health (Lond) ; 15: 1745506519864009, 2019.
Article in English | MEDLINE | ID: mdl-31378196

ABSTRACT

Hormone therapy use has undergone dramatic changes over the past 20 years. Widespread use of hormone therapy in the 1980s and 1990s came to an abrupt halt in the early 2000s after initial findings of the Women's Health Initiative trial were published and the study was terminated. Since then, much has been learned about the characteristics of women most likely to benefit from hormone therapy. There is general agreement that women younger than 60 years or who initiate hormone therapy within 10 years of menopause onset gain short-term benefit in terms of symptomatic relief and long-term benefit in terms of protection from chronic diseases that affect postmenopausal women. Despite accumulating evidence in support of hormone therapy for symptomatic menopausal women, the slow response by the medical community has led to a 'large and unnecessary burden of suffering' by women worldwide. Greater efforts are clearly needed to educate physicians and medical students about the pathophysiology of menopause and the role of hormone therapy in supporting women through the transition. This article provides a brief historical perspective of events that led to the backlash against hormone therapy, explores the current position of guideline groups, and provides practical recommendations to guide first-line management of symptomatic menopausal women.


Subject(s)
Estrogens/therapeutic use , Guidelines as Topic , Hormone Replacement Therapy/trends , Menopause/drug effects , Progestins/therapeutic use , Breast Neoplasms , Cardiovascular Diseases , Estrogen Replacement Therapy/trends , Female , Humans , Middle Aged , Risk Assessment
8.
Arch Anim Nutr ; 72(5): 396-406, 2018 Oct.
Article in English | MEDLINE | ID: mdl-30183391

ABSTRACT

We hypothesised that copper nanoparticles (NanoCu), because of their high physicochemical reactivity and bioavailability, could be used in much smaller quantities than bulk Cu, consequently reducing excretion of Cu into the environment. The objective of the study was to evaluate the effects of various levels of NanoCu on the development and growth of broiler chickens, in order to establish an optimum level of NanoCu dietary supplementation. Broiler chickens were randomly divided into five groups of 10 birds each. The control group received 7.5 mg Cu/kg feed (standard level) as CuSO4, while groups fed with complexes of NanoCu and starch received 25%, 50%, 75% and 100% of the standard level of Cu used in the control group. Chicken growth and excretion of Cu, Fe and Zn were measured during the growth period from d 7 to 42. At d 42, the slaughter characteristics, the content of Cu, Fe and Zn in the breast muscle and liver, and the oxidative status were analysed. The results indicate that using NanoCu can reduce the standard level of Cu from CuSO4 supplementation by 75% without jeopardising animal growth, and at the same time significantly decreasing Cu excretion into the environment.


Subject(s)
Chickens/metabolism , Copper/administration & dosage , Metal Nanoparticles/administration & dosage , 8-Hydroxy-2'-Deoxyguanosine , Analysis of Variance , Animal Feed/analysis , Animals , Chickens/growth & development , Colloids/chemistry , Copper/analysis , Copper/pharmacology , Copper Sulfate/administration & dosage , Deoxyguanosine/analogs & derivatives , Deoxyguanosine/analysis , Dose-Response Relationship, Drug , Feces/chemistry , Hydrogen-Ion Concentration , Iron/analysis , Liver/chemistry , Male , Metal Nanoparticles/ultrastructure , Microscopy, Electron, Transmission/veterinary , Minerals/administration & dosage , Oxidation-Reduction/drug effects , Pectoralis Muscles/chemistry , Powders , Random Allocation , Spectrophotometry, Atomic/veterinary , Thiobarbituric Acid Reactive Substances/analysis , Zinc/analysis
9.
Arch Anim Nutr ; 71(4): 327-332, 2017 Aug.
Article in English | MEDLINE | ID: mdl-28581358

ABSTRACT

The objective of this study was to evaluate bone resistance after in ovo administration of copper nanoparticles (NanoCu) and to determine the number of cells positive for proliferating cell nuclear antigen (PCNA) in the femoral bones of broiler chickens (n = 12 per group). The study demonstrated that femoral bones from the NanoCu group were characterised by a higher weight and volume and by significantly greater resistance to fractures compared to the Control group. NanoCu promoted the proliferation of PCNA-positive cells in the long bones of chickens. A significantly higher number of PCNA-positive cells in the bones of birds in the NanoCu group compared with the Control group (137 and 122, respectively) indicate a stimulatory effect during embryogenesis. Considering the improvement in bone resistance to fractures and the effect of NanoCu on the number of PCNA-positive cells in femoral bones, NanoCu may be an alternative agent to minimise the ever-present problem of weak bones in broiler chickens.


Subject(s)
Bone Density/drug effects , Chickens/physiology , Copper/pharmacology , Femur/drug effects , Metal Nanoparticles , Animals , Biomechanical Phenomena , Chick Embryo/growth & development , Femur/chemistry , Injections , Proliferating Cell Nuclear Antigen/metabolism
10.
J Sex Med ; 13(6): 887-904, 2016 06.
Article in English | MEDLINE | ID: mdl-27215685

ABSTRACT

Erectile dysfunction (ED) has been well recognized as a marker of increased cardiovascular risk for more than 15 years, especially in younger men. Early detection of ED represents an opportunity to intervene to decrease the risk of future cardiovascular events and limit the progression of ED severity. Evidence suggests there is a window of opportunity of 3 to 5 years from the onset of ED to subsequent cardiovascular events. This opportunity is usually missed if the onus is placed on the patient to seek care for his sexual problems. Unfortunately, these clear messages have not been incorporated into routine cardiovascular care. The reasons for these disparities within specialties are discussed in this article, in addition to management algorithms. Lifestyle modification is usually recommended as the first-line treatment to correct ED and lessen cardiovascular risk, but evidence suggests that this might be effective only in men without established cardiovascular comorbidities. In men with type 2 diabetes mellitus and established cardiovascular disease, lifestyle modification alone is unlikely to be effective. Cardiovascular medications are often associated with sexual dysfunction but changes in medication are more likely to be beneficial in men with milder recent-onset ED. A balanced view must be taken related to medication adverse events, taking into account optimal management of established cardiovascular disease. Testosterone deficiency has been associated with different metabolic disorders, especially metabolic syndrome and type 2 diabetes mellitus. Testosterone deficiency syndrome has been associated with an independent burden on sexual function globally and increased cardiovascular and all-cause mortality. Testosterone replacement therapy has been shown to improve multiple aspects of sexual function and, in some studies, has been associated with a decrease in mortality, especially in men with type 2 diabetes mellitus. Recent studies have suggested that phosphodiesterase type 5 inhibitors, the first-line medications to treat ED, could decrease cardiovascular and all-cause mortality, through multiple mechanisms, predominantly related to improved endothelial function.


Subject(s)
Cardiovascular Diseases/complications , Coronary Artery Disease/diagnosis , Diabetes Mellitus, Type 2/complications , Erectile Dysfunction/etiology , Comorbidity , Humans , Life Style , Male , Metabolic Syndrome/complications , Phosphodiesterase 5 Inhibitors/therapeutic use , Risk Factors , Sexuality , Testosterone/therapeutic use
11.
J Sex Med ; 13(4): 519-37, 2016 Apr.
Article in English | MEDLINE | ID: mdl-27045256

ABSTRACT

INTRODUCTION: The diagnosis, treatment, and survivorship of cancer have a profound effect on the quality of life and psychological well-being of men and women. Indeed, the perturbation of sexual function because of neoplasm has far-reaching implications. AIMS: To explore the prevalence, pathophysiology, and treatment of sexual issues in persons with cancer and offer evidence-based recommendations regarding optimal prevention and treatment strategies. METHODS: A committee of multidisciplinary specialists was formed as part of the larger International Sexual Medicine Consultation working with urologic and sexual medicine societies over a 1-year period to review the result of chronic-illness management on sexual function and satisfaction. The aims, goals, data collection techniques, and report format were defined by a central committee. MAIN OUTCOMES MEASURES: Expert consensus was based on evidence-based medical and psychosocial literature review, extensive group discussion, and an open presentation with a substantial discussion period. RESULTS: This summary evaluates contemporary literature concerning the prevalence, pathophysiology, and psychological impact of cancer diagnosis and treatment on sexual dysfunction. Evidence-based recommendations and guidelines for evaluation and management are presented. CONCLUSION: The diagnosis and treatment of cancer have a significant negative impact on sexual function and satisfaction. Comprehension of baseline sexual function, role of psychological supports, and available treatment options could attenuate the heavy burden of decreased sexual function.


Subject(s)
Neoplasms/complications , Neoplasms/psychology , Sexual Behavior/psychology , Sexual Dysfunction, Physiological/etiology , Sexual Dysfunctions, Psychological/etiology , Adult , Evidence-Based Practice , Female , Humans , Male , Middle Aged , Neoplasms/physiopathology , Personal Satisfaction , Practice Guidelines as Topic , Prevalence , Quality of Life , Sexual Dysfunction, Physiological/diagnosis , Sexual Dysfunction, Physiological/physiopathology , Sexual Dysfunctions, Psychological/diagnosis , Sexual Dysfunctions, Psychological/physiopathology , Survival Rate
12.
J Sci Food Agric ; 96(9): 3058-62, 2016 Jul.
Article in English | MEDLINE | ID: mdl-26417698

ABSTRACT

BACKGROUND: Copper (Cu) is a key trace mineral involved in a variety of physiological processes, and is commonly used in poultry production. However, regardless of the inclusion level the majority of Cu is excreted with poultry faeces. We hypothesise that in ovo administration will allow for better utilisation of Cu during embryo development than when supplied post-natally with feed to growing chickens. Thus, the objective of this study was to evaluate effects of in ovo administration of NanoCu and copper sulfate (CuSO4 ) on broiler chicken performance. RESULTS: The study showed the positive influences of Cu nanoparticles and CuSO4 on broiler chickens performance. Body weight, at the end of the rearing period (day 42) was significantly higher in NanoCu (2206 g) and CuSO4 (2402 g) groups compared to the control group (2000 g). Both treatment groups had significantly lower feed conversion rate and mortality, and higher percentage of breast and leg muscles in the carcass versus control. CONCLUSION: The in ovo application of Cu colloids may ensure an efficient penetration of Cu into the embryonic tissue with long lasting effects on postnatal growth. The method may provide a successful alternative to using Cu as a feed additive. © 2015 Society of Chemical Industry.


Subject(s)
Copper/administration & dosage , Embryonic Development , Food Quality , Meat/analysis , Metal Nanoparticles/administration & dosage , Muscle Development , Trace Elements/administration & dosage , Absorption, Physiological , Animals , Animals, Inbred Strains , Chick Embryo , Colloids , Cooking , Copper/chemistry , Copper/metabolism , Copper/toxicity , Copper Sulfate/administration & dosage , Copper Sulfate/chemistry , Copper Sulfate/metabolism , Copper Sulfate/toxicity , Denmark , Energy Intake , Environmental Pollution/prevention & control , Injections , Metal Nanoparticles/chemistry , Metal Nanoparticles/toxicity , Solubility , Trace Elements/chemistry , Trace Elements/metabolism , Trace Elements/toxicity , Water/analysis , Weight Gain
13.
Int J Mol Sci ; 16(3): 4838-49, 2015 Mar 03.
Article in English | MEDLINE | ID: mdl-25741768

ABSTRACT

Copper is a key element affecting blood vessel growth and muscle development. However, the ions released from Cu salts are toxic. Given their specific physicochemical properties, nanoparticles of Cu (NanoCu) may have different bioactivity and affect the development of blood vessel and muscles in a different manner than Cu salts. The objective of the study was to evaluate the influence of NanoCu on embryo development and angiogenesis at the systemic and molecular level, in experiments using a chick embryo model. Fertilized chicken eggs were divided into a control group, and groups injected with a placebo, CuSO4 or NanoCu. Embryo development at the whole body level and molecular indices using an embryo chorioallantoic membrane model were measured during embryogenesis. The present study indicated for the first time that NanoCu have pro-angiogenic properties at the systemic level, to a greater degree than CuSO4 salt. The properties of NanoCu were confirmed at the molecular level, demonstrating significant effects on mRNA concentration and on mRNA gene expression of all pro-angiogenic and pro-proliferative genes measured herein.


Subject(s)
Copper/chemistry , Embryonic Development/drug effects , Metal Nanoparticles/toxicity , Neovascularization, Physiologic/drug effects , Animals , Chick Embryo , Chickens , Chorioallantoic Membrane/blood supply , Copper Sulfate/chemistry , Copper Sulfate/toxicity , Metal Nanoparticles/chemistry , RNA, Messenger/metabolism , Vascular Endothelial Growth Factor A/genetics , Vascular Endothelial Growth Factor A/metabolism
14.
J Sci Food Agric ; 95(8): 1730-5, 2015 Jun.
Article in English | MEDLINE | ID: mdl-25155871

ABSTRACT

BACKGROUND: The purpose of this study was to determine the quality of meat and the histological structure of muscles of Ayam Cemani chickens, Ayam Cemani × Sussex hybrids and slow-growing Hubbard JA 957 chickens and to examine whether crossing generally available Sussex chickens with little available Ayam Cemani gives a good quality product of interest to the poultry industry and in food technology. RESULTS: The size of breast and leg muscle fibers varied among genotypes. The breast and leg muscles of slow-growing Hubbard JA 957 chickens had the largest fiber diameter. The histological and biochemical properties of muscles, including the type, number, proportions, diameter and metabolic profile of fibers, had a significant effect on the pH and water-binding capacity of meat, thus affecting its quality. The muscle fibers of Ayam Cemani chickens were approximately half the size of the muscle fibers of Hubbard JA 957 chickens. Ayam Cemani and Ayam Cemani × Sussex gave a product of as good quality as Hubbard JA 957 chickens. CONCLUSION: Meat from Ayam Cemani chickens is a rich source of protein and could be highly valued by gourmet consumers, connoisseurs and dieticians for its rarity and originality. The results of this study show that genotype (Ayam Cemani, Ayam Cemani × Sussex, Hubbard JA 957) affected the quality and color of meat and the histological profile of chicken breast and leg muscles.


Subject(s)
Chickens/genetics , Crosses, Genetic , Food Quality , Meat , Muscle, Skeletal/anatomy & histology , Animals , Breeding , Dietary Proteins/analysis , Extremities , Meat/analysis , Muscle Fibers, Skeletal , Pectoralis Muscles/anatomy & histology , Species Specificity
15.
J Sci Food Agric ; 95(12): 2562-5, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25348487

ABSTRACT

BACKGROUND: The aim of this study was to determine the effect of various doses of ß-alanine administered in feed mixtures and of heat treatment on changes in the contents of dipeptides anserine and carnosine in meat of fast-growing Hubbard Flex chickens. RESULTS: The study demonstrated a significant effect of dietary administration of ß-alanine on anserine content in breast muscles and on carnosine content in leg muscles of the chickens. A successive increase in dipeptide content was observed with increasing ß-alanine content in the feed mixture. As a result of heat treatment, a significant (P ≤ 0.01) increase was observed in the anserine content of both breast and leg muscles: over twofold higher anserine content was detected in heat-treated muscles compared with raw muscles. The study showed that interesting effects might occur during ß-alanine administration to feed mixtures for chickens, especially including increased contents of anserine and carnosine in skeletal muscles of the birds. CONCLUSION: Taking into account the well-documented health-promoting effect of histidine dipeptides and possibilities of increasing their contents in the body by supplementation with ß-alanine, anserine and carnosine might be considered as potential components of functional foods.


Subject(s)
Animal Feed/analysis , Chickens/growth & development , Dipeptides/metabolism , Muscle, Skeletal/metabolism , beta-Alanine/administration & dosage , Animals , Anserine/metabolism , Carnosine/metabolism , Chromatography, High Pressure Liquid , Dietary Supplements , Dose-Response Relationship, Drug , Female , Male
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