A preliminary discussion on carnosine dipeptidase 1 as a potential novel biomarker for the diagnostic and prognostic evaluation of hepatocellular carcinoma / 中华肝脏病杂志

Objective: To explore carnosine dipeptidase 1 (CNDP1) potential value as a diagnostic and prognostic evaluator of hepatocellular carcinoma (HCC). Methods: A gene chip and GO analysis were used to screen the candidate marker molecule CNDP1 for HCC diagnosis. 125 cases of HCC cancer tissues, 85 cases of paracancerous tissues, 125 cases of liver cirrhosis tissues, 32 cases of relatively normal liver tissue at the extreme end of hepatic hemangioma, 66 cases from serum samples of HCC, and 82 cases of non-HCC were collected. Real-time fluorescent quantitative PCR, immunohistochemistry, western blot, and enzyme-linked immunosorbent assay were used to detect the differences in mRNA and protein expression levels of CNDP1 in HCC tissue and serum. Receiver operating characteristic (ROC) curves and Kaplan-Meier survival were used to analyze and evaluate the value of CNDP1 in the diagnosis and prognosis of HCC patients. Results: The expression level of CNDP1 was significantly reduced in HCC cancer tissues. The levels of CNDP1 were significantly lower in the cancer tissues and serum of HCC patients than those in liver cirrhosis patients and normal controls. ROC curve analysis showed that the area under the curve of serum CNDP1 in the diagnosis of HCC patients was 0.753 2 (95% CI 0.676-0.830 5), and the sensitivity and specificity were 78.79% and 62.5%, respectively. The combined detection of serum CNDP1 and serum alpha-fetoprotein (AFP) significantly improved the diagnostic accuracy (AUC = 0.820 6, 95% CI 0.753 5-0.887 8). The diagnostic sensitivity and specificity of serum CNDP1 for AFP-negative HCC patients were 73.68% and 68.75% (AUC = 0.793 1, 95% CI 0.708 8-0.877 4), respectively. In addition, the level of serum CNDP1 distinguished small liver cancer (tumor diameter < 3 cm) (AUC = 0.757 1, 95% CI 0.637 4-0.876 8). Kaplan-Meier survival analysis showed that CNDP1 was associated with a poor prognosis in HCC patients. Conclusion: CNDP1 may be a potential biomarker for the diagnostic and prognostic evaluation of HCC, and it has certain complementarity with serum AFP.

Mitochondrial energy metabolism in baby hamster kidney (BHK-21/C13) cells treated with karnozin extra® / Metabolismo energético mitocondrial en células de riñón de hámster bebé (BHK-21 / C13) tratadas con karnozin extra®

SUMMARY: Carnosine is known as a natural dipeptide, which inhibits the proliferation of tumor cells throughout its action on mitochondrial respiration and cell glycolysis. However, not much is known about its effects on the metabolism of healthy cells. We explored the effects of Karnozin EXTRA® capsule with different concentrations of L-carnosine, on the cell viability and the expressions of intermediate filament vimentin (VIM) and superoxide dismutase (SOD2) in normal fibroblasts BHK-21/C13. Furthermore, we investigated its action on the energy production of these cells. Cell viability was quantified by the MTT assay. The Clark oxygen electrode (Oxygraph, Hansatech Instruments, England) was used to measure the "intact cell respiration rate", state 3 of ADP-stimulated oxidation, maximum oxidation capacity and the activities of complexes I, II and IV. Results showed that Karnozin EXTRA® capsule in concentrations of 2 and 5 mM of L-carnosine did not induce toxic effects and morphological changes in treated cells. Our data revealed a dose-dependent immunofluorescent signal amplification of VIM and SOD2 in the BHK-21/C13 cell line. This supplement substantially increased the recorded mitochondrial respiration rates in the examined cell line. Due to the stimulation of mitochondrial energy production in normal fibroblasts, our results suggested that Karnozin EXTRA® is a potentially protective dietary supplement in the prevention of diseases with altered mitochondrial function.

RESUMEN: La carnosina se conoce como dipéptido natural, que inhibe la proliferación de células tumorales a través de su acción sobre la respiración mitocondrial y la glucólisis celular. Sin embargo, no se sabe mucho de sus efectos sobre el metabolismo de las células sanas. Exploramos los efectos de la cápsula Karnozin EXTRA® con diferentes concentraciones de L-carnosina, sobre la viabilidad celular y las expresiones de vimentina de filamento intermedio (VIM) y superóxido dismutasa (SOD2) en fibroblastos normales BHK-21 / C13. Además, estudiamos su acción sobre la producción de energía de estas células. La viabilidad celular se cuantificó mediante el ensayo MTT. Se utilizó el electrodo de oxígeno Clark (Oxygraph, Hansatech Instruments, Inglaterra) para medir la "tasa de respiración de células intactas", el estado 3 de oxidación estimulada por ADP, la capacidad máxima de oxidación y las actividades de los complejos I, II y IV. Los resultados mostraron que la cápsula de Karnozin EXTRA® en concentraciones de 2 y 5 mM de L- carnosina no indujo efectos tóxicos ni cambios morfológicos en las células tratadas. Nuestros datos revelaron una amplificación de señal inmunofluorescente dependiente de la dosis de VIM y SOD2 en la línea celular BHK-21 / C13. Este suplemento aumentó sustancialmente las tasas de respiración mitocondrial registradas en la línea celular examinada. Debido a la estimulación de la producción de energía mitocondrial en fibroblastos normales, nuestros resultados sugirieron que Karnozin EXTRA® es un suplemento dietético potencialmente protector en la prevención de enfermedades con función mitocondrial alterada.

Investigação dos processos biológicos associados à produção endógena de aldeídos reativos / Research on biological processes associated with endogenous production of reactive aldehydes

Os aldeídos são espécies reativas que podem ser produzidos endogenamente por processos como a lipoperoxidação, podendo reagir com lipídios, proteínas e DNA. Diversas evidências apontam para o envolvimento de aldeídos reativos na progressão de patologias como doenças cardiovasculares, arteriosclerose e doenças neurodegenerativas. Uma meta central do CEPIDRedoxoma é estudar a reatividade química de intermediários redox em ambientes biológicos e consequentes mudanças na estrutura e função de biomoléculas, entender como cada intermediário redox reage com biomoléculas específicas e os efeitos resultantes, essenciais para a concepção de biomarcadores e antioxidantes. O nosso grupo estuda os mecanismos de formação, detoxificação e reação com biomoléculas de aldeídos reativos endógenos e exógenos e seu papel em patologias como a esclerose lateral amiotrófica (ALS). Um dos mecanismos de detoxificação desses aldeídos é através da conjugação com a carnosina. Recentemente, foi observado que a suplementação de animais transgênicos ALS SOD G93A com carnosina via oral resultou em retardo da perda de peso e tendência de aumento da sobrevida dos animais. O presente projeto buscou investigar o possível papel da carnosina em animais modelo para ALS. Para isso as modificações em DNA induzidas por aldeídos reativos e a formação de adutos de carnosina-aldeídos foram analisadas através de metodologia HPLC-MS/MS. Assim observamos que ratos suplementados com carnosina apresentaram níveis significativamente menores de proteína carbonilada em músculo e fígado. Em fígadoforam vistos níveis menores de dois adutos de DNA, 8-oxodGuo e1,N2-HO-propanodGuo, em animais suplementados. Em cérebro foram detectados níveis menores de 1, N2-εdGuo. Com relação aos adutos carnosina-aldeídos, foi observado níveis significativamente maiores do aduto CAR-HHE na medula. Com embasamento nos resultados aqui apresentados, sugere-se a utilização de sequestradores de aldeídos como uma estratégia terapêutica em condições fisiopatológicas nas quais ao acúmulo dessas espécies está comprovado

Aldehydes are reactive species that can be produced endogenously by processes such as lipid peroxidation, which can react with lipids, proteins and DNA. Several evidences point to the involvement of reactive aldehydes in the progression of pathologies such as cardiovascular diseases, atherosclerosis and neurodegenerative diseases. A central goal of CEPID-Redoxoma is to study the chemical reactivity of redox intermediates in biological environments and consequent changes in the structure and function of biomolecules, to understand how each redox intermediate reacts with specific biomolecules and the resulting effects, essential for the design of biomarkers and antioxidants. Our group studies the mechanisms of formation, detoxification and reaction with biomolecules of endogenous and exogenous reactive aldehydes and their role in pathologies such as amyotrophic lateral sclerosis (ALS). One of the detoxification mechanisms of these aldehydes is through carnosine conjugation. Recently, we observed that oral carnosine supplementation in transgenic ALS SODG93A animals resulted in delayed weight loss and a tendency to increase the survival of the animals. The present project investigated the potential role of carnosine in animal models for ALS. Thus, reactive aldehydes induced DNA modifications and carnosine aldehyde adducts were analyzed by HPLC-MS/MS. We observed that rats supplemented with carnosine presented significantly lower levels of protein carbonylation in muscle and liver. Lower levels of two DNA adducts, 8-oxodGuo and 1, N2-HO-propanodGuo, were observed in liver of the supplemented animals. Lower levels of 1, N2-εdGuo were detected in the brain. Regarding the carnosine-aldehydeadducts, significantly higher levels of the CAR-HHE adduct were observed in spinal cord. The results presented here suggest the use of aldehyde scavengers as a therapeutic strategy under pathological conditions in which is proven the accumulation of these species

Effect of ß-alanine supplementation on 20 km cycling time trial performance

The effects of ß-alanine supplementation on high-intensity cycling performance and capacity have been evaluated, although the effects on longer duration cycling performance are unclear. Nineteen UK category 1 male cyclists completed four 20 km cycling time trials, two before and two after supplementation with either 6.4 g•d-1 ß-alanine (n = 10; BA) or a matched placebo (n = 9; P). Performance time for the 20 km time trial and 1 km split times were recorded. There was no significant effect of ß-alanine supplementation on 20 km time trial performance (BA-pre 1943 ± 129 s; BA-post 1950 ± 147 s; P-pre 1989 ± 106 s; P-post 1986 ± 115 s) or on the performance of each 1 km split. The effect of ß-alanine on 20 km time trial performance was deemed unclear as determined by magnitude based inferences. Supplementation with 6.4 g•d-1 of ß-alanine for 4 weeks did not affect 20 km cycling time trial performance in well trained male cyclists

Biological functions of histidine-dipeptides and metabolic syndrome

The rapid increase in the prevalence of metabolic syndrome, which is associated with a state of elevated systemic oxidative stress and inflammation, is expected to cause future increases in the prevalence of diabetes and cardiovascular diseases. Oxidation of polyunsaturated fatty acids and sugars produces reactive carbonyl species, which, due to their electrophilic nature, react with the nucleophilic sites of certain amino acids. This leads to formation of protein adducts such as advanced glycoxidation/lipoxidation end products (AGEs/ALEs), resulting in cellular dysfunction. Therefore, an effective reactive carbonyl species and AGEs/ALEs sequestering agent may be able to prevent such cellular dysfunction. There is accumulating evidence that histidine containing dipeptides such as carnosine (beta-alanyl-L-histidine) and anserine (beta-alanyl-methyl-L-histidine) detoxify cytotoxic reactive carbonyls by forming unreactive adducts and are able to reverse glycated protein. In this review, 1) reaction mechanism of oxidative stress and certain chronic diseases, 2) interrelation between oxidative stress and inflammation, 3) effective reactive carbonyl species and AGEs/ALEs sequestering actions of histidine-dipeptides and their metabolism, 4) effects of carnosinase encoding gene on the effectiveness of histidine-dipeptides, and 5) protective effects of histidine-dipeptides against progression of metabolic syndrome are discussed. Overall, this review highlights the potential beneficial effects of histidine-dipeptides against metabolic syndrome. Randomized controlled human studies may provide essential information regarding whether histidine-dipeptides attenuate metabolic syndrome in humans.

The Neuroprotective Effects of Carnosine in Early Stage of Focal Ischemia Rodent Model

OBJECTIVE: This study was conducted to elucidate neuroprotective effect of carnosine in early stage of stroke. METHODS: Early stage of rodent stroke model and neuroblastoma chemical hypoxia model was established by middle cerebral artery occlusion and antimycin A. Neuroprotective effect of carnosine was investigated with 100, 250, and 500 mg of carnosine treatment. And antioxidant expression was analyzed by enzyme linked immunosorbent assay (ELISA) and western blot in brain and blood. RESULTS: Intraperitoneal injection of 500 mg carnosine induced significant decrease of infarct volume and expansion of penumbra (p<0.05). The expression of superoxide dismutase (SOD) showed significant increase than in saline group in blood and brain (p<0.05). In the analysis of chemical hypoxia, carnosine induced increase of neuronal cell viability and decrease of reactive oxygen species (ROS) production. CONCLUSION: Carnosine has neuroprotective property which was related to antioxidant capacity in early stage of stroke. And, the oxidative stress should be considered one of major factor in early ischemic stroke.

beta-alanyl-l-histidine, an anti-oxidant, anti-fibrotic and geno/cytotoxicity inhibitor: a sheild against cc14-induced hepatotoxicity in rats

Hepatic fibrosis is a consequence of chronic liver injury and is characterized by an excessive hepatic connective tissue formation and deposition. p-alanyl-L-histidine [carnosine] is a dipeptide with anti-inflammatory and antioxidant properties in addition to its involvement in many physiological process. Specific treatments to stop progressive liver fibrosis are not available, so that the objective of the present study was to evaluate the hepatoprotective, antioxidant, antifibrotic, cyto/genoprotective effect of p-alanyl-L-histidine against hepatic injury induced by carbon tetrachloride [CCU] in rats. p-alanyl-L-histidine showed significant protection with normalization of liver aminotransferases, increased glutathione S transferase [GST] activity while decreased hepatic hydroxyproline, protein carbonyl, hydrogen peroxide[H[2]O[2]] levels, DNA damage, Cytochrome P450[2]Ei [CYP[2]Ei] activity and transforming growth factor-pi [TGF-pl] mRNA level. In conclusion: P-alanyl-L-histidine possesses hepatoprotective properties through reducing hepatic toxicity markers, oxidative stress, cytotoxicity, genotoxicity, fibrosis and improvement of the histological architecture of the liver

CCl[4] ilnduced hepatotoxicity: protective effects of carnosine on peroxisome proliferator-activated receptor gamma gene expression, genotoxicity and oxidant/antioxidant status in rats

Effect of carnosine as an antioxidant in protection against carbon tetrachloride CCI[4] induced oxidative stress and hepatotoxicity in rats was investigated. Liver toxicity was induced in rat model at which four experimental groups of 20 rats each were constructed: group [1] the control group in which rats were not administrated CCI[4] or carnosine; group [II] CCI[4] group in which rats were subcutaneously injected with CCI[4] in a dose of 2 ml /Kg body weight twice weekly for 4 weeks; group[III] CCI[4] and carnosine group in which rats were also subcutaneously injected with CCI[4] and co-treated with daily intraperitoneal [i.p.] carnosine at a dose of 10 mg / kg body weight and group [IV] received also i.p. repeated daily dose of carnosine. Hepatotoxicity was assessed by measurement of serum aspartate aminotransferase [AST] and alanine aminotransferase [ALT] activities. Hepatic peroxisome proliferator-activated receptor gamma [PPARgammay] mRNA expression, glutathione-S-transferase [GST] activity, paraoxonase 1 [PON1] activity, xantheine oxidase [XO] activity and total anti-oxidant capacity [TAC] level as well as DNA damage in blood were evaluated. The results were confirmed by histopathological examination. Carnosine treatment significantly prevented the CCI[4] induced hepatotoxicity, oxidative stress and DNA damage. In conclusion, our results suggested that carnosine might be a therapeutic antifibrotic/antigenotoxic agent for the treatment of CCI[4-] induced hepatotoxicity due to its antioxidant properties

Protective effects of carnosine against closed head injury in mice / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the protective effects of carnosine against experimental closed head injury (CHI) in mice.</p><p><b>METHODS</b>The CHI model was established by free-falling weight-drop. Carnosine (250 mg/kg or 500 mg/kg) was administered intraperitoneally 30 min before brain trauma, then q.d for 7 d; while normal saline was administrated for control group. The neurological defect was evaluated by neurological severity score (NSS) within 7 d; the survival rate and the histological alternations were observed.</p><p><b>RESULTS</b>Carnosine prevented the body weight loss of mice at dose of 500 mg/kg; significantly increased the survival rate, and reduced the neurological defect and histological damage at dose of 250 and 500 mg/kg.</p><p><b>CONCLUSION</b>Carnosine can attenuate closed head injury in mice.</p>

Protective effect of carnosine on the injury of rat vascular endothelial cells induced by hypoxia / 中国应用生理学杂志

<p><b>OBJECTIVE</b>To investigate the effect of camosine on the injury of rat vascular endothelial cells(VECs) induced by hypoxia.</p><p><b>METHODS</b>The model of the injury of rat VECs induced by hypoxia was established. The effect of camosine on injury of VECs activity induced by hypoxia was determined by MTT assay. The levels of lactate dehydrogenase (LDH) activity in cell medium were measured with corresponding kit. The cell structure was observed under microscope after Coomassie brilliant blue R-250 staining.</p><p><b>RESULTS</b>After culturing VECs with camosine (10 to 20 mmol/L) for 6 hours, the decrease in VECs activity induced by 12 and 24 hour hypoxia was inhibited. The release of LDH was also inhibited, and the integrity of cell structure remained.</p><p><b>CONCLUSION</b>Camosine has the protective effect on hypoxia injured VECs.</p>

Enzymetic synthesis and characterization of a carnosine analogue in non-aqueous solvent / 生物工程学报

Carnosine (beta-Ala-L-His) has high antioxidant activity, and it is widely used in biology, chemical engineering, medicine and other fields. Its analogue syntheised in non-aqueous solvent and catalyzed by enzymes is high-effective but low-price, so it has great prospect. Here, we synthesized a carnosine analogue imidazole 4(5)-alanylamide-5(4)-carboxylic acid with imidazole-4,5-dicarboxylic acid and L-Alanine as substrates, alpha-chymotrypsin as catalyst in tetrahydrofuran (THF) solvent. Based on the orthogonal experiments, the optimized synthetic conditions are 4,5-dicarboxylic acid: L-alanine = 1:3 (m/m), alpha-chymotrypsin: substrates (4,5-dicarboxyl acid and L-alanine) = 1:200 (m/m), pH 8 phosphate buffer:THF = 1.6:10 (V/V), reaction temperature 35 degrees C, time 1.5 h. We separated the product with silica gel G60 thin-layer chromatography (TLC), and a new spot appeared at Rf (ratio to front) = 0.81; then the new spot was purified and characterized with UV spectra, high performance liquid chromatogram (HPLC) and 13C NMR (13C nuclear magnetic resonance). The UV spectra shows a new absorption peak at 310 nm, and the peak in 253 nm is largely strengthened; HPLC reserve times are all 4.5 min at 253 nm, 310 nm, 330 nm; 13C NMR shows 8 carbons. Combing with the catalytic mechanism of alpha-chymotrypsin, structure of the analogue is confirmed, i.e., imidazole 4(5)-alanylamide-5(4)-carboxylic acid.

Carnosine and diseases of central nervous system / 浙江大学学报·医学版

The naturally-occurring dipeptide carnosine (beta-alanyl-L-histidine) is found exclusively in animal tissues, such as brain and skeletal muscle tissues. Carnosine is a potent hydrophilic antioxidant, antiglycating agent, reactive oxygen species scavenger and pH-buffer. Recent reports suggest that carnosine has potential therapeutic applications in many diseases of central nervous system, such as Alzheimer's disease, Parkinson's disease and cerebral ischemic diseases. To investigate the relationship between carnosine and diseases of central nervous system, and to research and develop carnosine drugs will shed light on a new way for treatment of diseases of central nervous system.

When can carnosine be the magic stick for lead-induced tissue damage? A toxicological biochemical histological study on albino rats

This study aimed at the assessment of the possible level of defence line of carnosine in chronic lead exposure. Evaluation of the possible relation of antioxidants and other functioning enzymes versus chronic lead exposure is also to be percussed. For this purpose, the study included two parts: Part [1]: was performed to deduce the correlation between the dose of carnosine and the different biochemical parameters measured in the studied tissues; Part [2]: was a long term study in which [56] male Wistar albino rats, divided into [6] groups, eight in each group: Group [1]: Negative control; Group [2]: Rats were treated with carnosine in a dose of 0.05 mg/g rat; Group [3]: Rats were treated with carnosine in a dose of 0.12 mg/ g rat; Group [4]: Comprised rats treated with lead acetate in a dose of [0.01 mg/g rat]; Group [5]: Included rats treated with lead [0.01 mg/g rat] and carnosine [0.05 mg/g rat]; Group [6]: Included rats treated with lead [0.01 mg/g rat] and carnosine [0.12 mg/g rat]. Treatment was performed daily for 6 days/ week for 5 months. The neural [brain] and visceral [heart, kidneys and liver] tissues were collected and different biochemical variables were determined; as well as a histological examination. In all the studied tissues, carnosine would only act as a free radical scavenger in the low dose only; and it could be used as an adjuvant in a low dose with chelating agents, since it cannot chelate lead in the brain tissue. Renal impairment was most probably a direct action of carnosine in a high dose on renal tissue, rather than being a result of lead intoxication. On the other hand, when carnosine was given in a small dose, it acted as a free radical scavenger. In the liver, carnosine in a small dose could not act as a chelator for lead, while in a high dose, it acts as a chelating agent. On the contrary, in a small dose, carnosine acts as a free radical scavenger but not in the high dose. Overall, carnosine cannot prevent the production of free radicals but can scavenge them only in a small dose

Carnosine protects against gentamicin-induced nephrotoxicity in rats

Acute renal failure is a major complication of gentamicin, limiting use of this antibiotic in treatment of gram negative infections. Reactive oxygen species are hypothesized to be a major factor in the nephrotoxicity of gentamicin and measures controlling this oxidative damage are widely appreciated. This work was conducted to test the hypothesis that treatment with carnosine, a biological antioxidant, may prevent or ameliorate acute renal injury, using a rat model of gentamicin-induced nephrotoxicity. Male wistar albino rats were assigned to one of six treatment groups; group I [control] rats were given normal saline injections daily for 10 days; group II rats were given IM gentamicin injections, 100 mg/kg/day, for 6 days: group III, IV and V rats were given gentamicin, together with IP carnosine injections 50, 100 and 200 mg/kg/day, respectively, for 10 days starting 4 days before gentamicin injections; and group VI rats were given only carnosine 200 mg/kg/day, for 10 days. All rats were weighed before and after experimentation, and 24 hour urine volume were collected in metabolic cages. At end of study, blood samples were collected for measurement of BUN, creatinine level and creatinine clearance. Rats were then sacrificed and the kidneys were excised. The left kidneys were homogenized and used for biochemical determination of MDA, GPX and SOD, while the right kidneys were processed for histological examination and scoring of renal cortical pathology. Results showed that gentamicin produced evident nephrotoxic effects revealed by; increased kidney weight, increased urine volume, elevations of serum levels of BUN and creatinine and decreased creatinine clearance; together with increased MDA, reduced GPX and SOD in kidney tissues. Marked histological alterations were also evident in the renal cortex [acute tubular necrosis of grade 2-3]. Carnosine treatment leads to significant dose-related attenuation of nephrotoxic effects of gentamicin revealed by reduction of the elevated biochemical parameters, improved oxidative status in the kidney, and amelioration of the histological changes. It is concluded that carnosine treatment could ameliorate the severity of renal cortical necrosis induced by gentamicin and maintain a better renal function. Thus, carnosine may be a useful candidate in the combination therapy with gentamicin to limit free radical-mediated renal injury

Avaliação da influência do dipeptídeo N-Beta-alanil-L-histidina (L-carnosina) sobre a cinética de expansão de culturas de células diplóides humanas, estirpe MRC-5 / Evaluation of the influence of dipeptide N-Beta-alanyl-L-histidine (L-carnosine) on the kinetics of growth of cultures of human diploid cell strain MRC-5

Um acordo internacional de transferência de tecnologia, firmado em 2003, entre a FIOCRUZ e a GlaxoSmithKline permitiu o acesso de Bio-Manguinhos a um moderno processo produtivo da vacina tríplice viral (TVV) contra o sarampo, a caxumba e a rubéola. A produção dessa vacina é parte do compromisso de Bio-Manguinhos com o programa nacional de auto-suficiência em imunobiológicos, tido como uma meta prioritária da política de saúde pública do governo brasileiro. A tecnologia aplicada para a produção da TVV envolve o uso de células MRC-5 como substrato celular para a produção de vírus vacinais da rubéola, particularmente da cepa Wistar RA27/3. A estirpe MRC-5 é reconhecida como um dos mais importantes substratos celulares para a produção de vacinas virais, e também sido adotada como modelo de estudo para senescência celular in vitro. A senescência celular é um estágio fisiológico complexo pelo qual, invariavelmente, qualquer população de células somáticas normais passa após atingir um determinado número de mitoses. Esse estágio fisiológico é caracterizado nas células diplóides pela contenção da capacidade de se multiplicar e pelo desenvolvimento de alterações morfológicas peculiares, especialmente quando cultivadas in vitro. Com o objetivo de aprimorar o monitoramento de estirpes de células diplóides humanas (HDCS - do inglês Human Diplóide Cells Strains) e contribuir para o estabelecimento da base de conhecimento necessário para a futura aplicação no processo de produção de TVV em Bio-Manguinhos, nós avaliamos culturas de células MRC-5 condicionadas com carnosina, em três diferentes aspectos: cinética de crescimento, propagação da cepa vacinal, Wistar RA27/3, do vírus da rubéola e a expressão do marcador biológico de senescência, a enzima Beta-galactosidase AS (Beta-gal AS). A avaliação do potencial antioxidante e antisenescente atribuído a carnosina, um dipeptídeo ubíquo à fisiologia de todos os animais superiores, sobre as células MRC-5 pode contribuir...

Evaluation of radioprotective effect of carnosine (beta- alanyl-1- histidine) on the wound healing in rats

O objetivo deste estudo foi avaliar o efeito radioprotetor da carnosina (b-alanil-1-histidina) no processo de cicatrizacão em ratos. Para isto, 48 ratos machos foram submetidos a um procedimento cirúrgico para realizacão de uma ferida retangular na região dorsal anterior. Os animais foram divididos aleatoriamente em 4 grupos experimentais: controle, irradiado, carnosina irradiado e carnosina. Os grupos carnosina e carnosina irradiado foram exposto a uma dose de corpo todo de 6 Gy de radiacão gama, 72 horas após a cirurgia para confeccão da ferida. O grupo carnosina e carnosina irradiado, adicionalmente, ao procedimento cirúrgico e a irradiacão, receberam duas doses de solucão aquosa de carnosina, a primeira sendo injetada 48 horas após a cirurgia, e a segunda 1 hora e 30 minutos antes da irradiacão. A reparacão tecidual nos 4 grupos foi avaliada 4, 7, 14 e 21 dias após a realizacão da ferida, por métodos morfológicos, histoquímicos e histofísicos. Em todos os períodos examinados, pode ser observado que os animais do grupo carnosina irradiado apresentaram um melhor desenvolvimento do tecido de granulacão do que o grupo irradiado e muito similar ao do grupo controle. Logo, de acordo com as condicões experimentais usadas, foi possível concluir que a carnosina é uma substância radioprotetora efetiva.

The induction apoptosis of HL-60 cells by low molecular weight compounds of taurine, ornithine and carnosine from new born calf liver / 中国应用生理学杂志

<p><b>AIM</b>Clinical studies stated that low molecular weight compounds (< 1.0 kd) extracted from the new born calf liver could effectively inhibit the proliferation of tumor cells. In this report, we observed inhibition effects and their regulative mechanisms of taurine, ornithine, carnosine on the proliferation of HL-60 cells.</p><p><b>METHODS</b>Three active ingredients, i.e., taurine, ornithine and carnosine were separated by ion-exchange chromatographic column and identified from the low molecular weight filtrate of new born calf liver. MTT assay was used to test the survival rate of HL-60 cells and normal lymphocytes treated by the three ingredients. The various effects of the three compounds on HL-60 cells were respectively evaluated by agarose gel electrophoresis, ESR and immunohistochemical methods.</p><p><b>RESULTS</b>These compounds effectively inhibited the proliferation of HL-60 cells and induced apoptosis which was determined by apoptotic changes in morphology and nuclear DNA degradation. Whereas no inhibition effects on normal lymphocytes were observed. In addition, the results of ESR showed that the activity of oxygen radical within HL-60 cells treated with there compounds decreased to trace level. Furthermore, in the immunohistochemical experiments, we found that the level of p45/skp2 in HL-60 cells decreased while the level of p27/kip increased.</p><p><b>CONCLUSION</b>The taurine, ornithine and carnosine compounds can selectively suppress tumor cells proliferation by regulating the level of cell cycle proteins.</p>

Hereditary non-syndromic hearing loss: influence of different variables

Non-syndromic hearing loss [NSHL] was studied in twenty-five patients using a clinical, audiological, cytogenetic and neurobiochemical evaluation. The study group was divided into five subgroups according to severity of hearing loss. Positive parental consanguinity was present in 84% of cases and similarly affected family members were present in 76%. All patients had no congenital malformations and were not dysmorphic. Patients possibly exposed to environmental factors were excluded from the study. Abnormal karyotyping was present in three cases, one case showed chromosome 15p 4, another case showed chromosomal del 11q22.1 and in the third case [47,XY] there was marker chromosome 15. Fluorescence in situ Hybridization [FISH] technique was performed on the case which showed marker. The study group showed significant lowering of five plasma amino acid levels [glutamic acid, aspartic acid, histidine, 3-methylhistidine and carnosine]. There was significant correlation between severity of hearing loss and each of the following: patient's age, glutamic acid, aspartic acid, 3-methylhistidine and carnosine. Identification of NSHL early after birth, as well as, amino acid screening is essential, to allow for faster therapeutic intervention and proper genetic counseling

Biochemical modifications induced in rabbits by schistosoma mansoni antigens and the beneficial effect of carnosine treatment

This immunological study involved individual injection of the three Schistosoma mansoni antigens [Ags], soluble egg antigen [SEA], cercarial antigen preparation [CAP] or soluble worm antigen preparation [SWAP] in three rabbits groups [Ag], respectively. Three other groups each received the same specific antigen conjoined with administration of L-carnosine [Ag-C] were also included in this study. Determination of three hepatic parameters and ten serum proteins was done. These were total protein, glycogen content and glycogen phosphorylase b activity of liver as well as serum total protein and nine protein fractions [alpha 2-macrglobulin, beta galactosidase, phosphorylase b, serum albumin, fumarase, carbonic anhydrase, beta- lactoglobulin, alpha-lactalbumin and aprotinin]. Conjoined carnosine treatment produced numerous variations. SEA-I-C group presented sex decreased parameters. In CAP-I-C animals, hepatic glycogen content was increased; while phosphorylase b activity was decreased as well as the concentration of serum parameters, total serum protein, alpha2-macroglobulin, phosphorylase b, albumin, fumarase, carbonic anhydrase, alpha-lactalbumin and aprotinin. In SWAP-I-C group, the concentration of only one fraction was decreased; carbonic anhydrase. In batch A, both the Ags. of the egg and cercaria, developmental stages having transient residence in the animal host, showed more affection by the specific Ag. Although, carnosine modified the results of all the three groups in batch B, its effect on both the egg and cercaria Ags. was still more than that of worm

Effect of alahistidine on brain histamine content and seizure development / 浙江大学学报·医学版

<p><b>OBJECTIVE</b>To investigate the effect of alahistidine on brain histamine content and seizure development.</p><p><b>METHODS</b>The kindling seizure was induced by ip injection with subconvulsant dose of pentylenetetrazole every 48 h. Monoamines and their metabolites were measured using a HPLC system and fluorometric assay.</p><p><b>RESULT</b>Chronic low histamine feeding markedly decreased histamine content in cortex and hypothalamus, and promoted seizure development induced by pentylenetetrazole. However, alahistidine feed reversed the decreased histamine content and slowed seizure development caused by low histamine feed. Both low histamine and alahistidine feed had no effect on norepinephrine, dopamine and its metabolites.</p><p><b>CONCLUSION</b>Alahistidine may affect histaminergic system and seizure development.</p>