RESUMO
The balance between protein synthesis and degradation regulates the amount of expressed proteins. This protein turnover is usually quantified as the protein half-life time. Several studies suggest that protein degradation decreases with age and leads to increased deposits of damaged and non-functional proteins. Glycation is an age-dependent, non-enzymatic process leading to posttranslational modifications, so-called advanced glycation endproducts (AGE), which usually damage proteins and lead to protein aggregation. AGE are formed by the Maillard reaction, where carbonyls of carbohydrates or metabolites react with amino groups of proteins. In this study, we quantified the half-life time of two important receptors of the immunoglobulin superfamily, the neural cell adhesion molecule (NCAM) and the receptor for advanced glycation end products (RAGE) before and after glycation. We found, that in two rat PC12 cell lines glycation leads to increased turnover, meaning that glycated, AGE-modified proteins are degraded faster than non-glycated proteins. NCAM is the most prominent carrier of a unique enzymatic posttranslational modification, the polysialylation. Using two PC12 cell lines (a non-polysialylated and a polysialylated one), we could additionally demonstrate, that polysialylation of NCAM has an impact on its turnover and that it significantly increases its half-life time.
Assuntos
Produtos Finais de Glicação Avançada/metabolismo , Ácido N-Acetilneuramínico/metabolismo , Moléculas de Adesão de Célula Nervosa/metabolismo , Receptor para Produtos Finais de Glicação Avançada/metabolismo , Animais , Glicosilação , Meia-Vida , Células PC12 , RatosRESUMO
OBJECTIVE: To describe the genetic characteristics and the management of two very rare cases of unilateral multifocal inner ear and internal auditory canal or cerebellopontine angle cochleovestibular schwannomas not being associated to full neurofibromatosis type 2-related schwannomatosis. PATIENTS: In a 29-year-old man and a 55-year-old woman with single-sided deafness multifocal unilateral cochleovestibular schwannomas were surgically resected, and hearing was rehabilitated with a cochlear implant (CI). Unaffected tissue was analyzed using next generation sequencing of the NF2 gene. Tumor tissue was analyzed using a 340-parallel sequencing gene panel. MAIN OUTCOME MEASURES: Mutations in the NF2 gene, word recognition score for monosyllables at 65 dB SPL (WRS 65 ) with CI. RESULTS: No disease-causing mutation was detected in the examined sequences in blood leucokytes. All tumor samples revealed, among others, somatic pathogenic NF2 mutations. While the anatomically separate tumors in case 1 were likely molecular identical, the tumors in case 2 showed different genetic patterns. WRS 65 was 55% at 6 years of follow-up and 60% at 4.5 years of follow-up, respectively. CONCLUSIONS: The occurrence of multifocal unilateral cochleovestibular schwannomas without pathogenic variants in NF2 in non-affected blood leucocytes can be associated with mosaic NF2 -related schwannomatosis (case 1), or with likely sporadic mutations (case 2) and may be overlooked due to their extreme rarity. Although challenging, successful hearing rehabilitation could be achieved through surgical resection of the tumors and cochlear implantation.
Assuntos
Ângulo Cerebelopontino , Implante Coclear , Neuroma Acústico , Humanos , Feminino , Pessoa de Meia-Idade , Implante Coclear/métodos , Masculino , Adulto , Neuroma Acústico/cirurgia , Neuroma Acústico/genética , Neuroma Acústico/patologia , Ângulo Cerebelopontino/cirurgia , Ângulo Cerebelopontino/patologia , Orelha Interna/cirurgia , Orelha Interna/patologia , Neurilemoma/cirurgia , Neurilemoma/genética , Neurilemoma/patologia , Mutação , Neoplasias da Orelha/cirurgia , Neoplasias da Orelha/genética , Neoplasias da Orelha/patologia , Neurofibromina 2/genéticaRESUMO
One hallmark of molecular aging is glycation, better known as formation of so-called advanced glycation end products (AGEs), where reactive carbonyls react with amino-groups of proteins. AGEs accumulate over time and are responsible for various age-dependent diseases and impairments. Two very potent dicarbonyls to generate AGEs are glyoxal (GO) and methylglyoxal (MGO). The plasma level of such dicarbonyls is higher in aging and age-related diseases. Natural killer (NK) cells are cells of the innate immune system and provide a major defense against tumor cells and virus infected cells. They are able to kill modified or infected cells and produce different cytokines to modulate the function of other immune cells. Here we investigated the effect of GO- and MGO-induced glycation on the function of NK cells. Using the human NK cell line NK-92, we could demonstrate that both GO and MGO lead to glycation of cellular proteins, but that MGO interferes much stronger with NK cell function (cytotoxicity) than GO. In addition, glycation of NK cell targets, such as K562 tumor cells, also interferes with their lysis by NK cells. From this data we conclude that glycation acts negatively on NK cells function and reduces their cytotoxic potential towards tumor cells.
Assuntos
Citotoxicidade Imunológica , Produtos Finais de Glicação Avançada/metabolismo , Células Matadoras Naturais/imunologia , Envelhecimento/imunologia , Apoptose/efeitos dos fármacos , Citotoxicidade Imunológica/efeitos dos fármacos , Glioxal/farmacologia , Humanos , Células K562 , Células Matadoras Naturais/efeitos dos fármacos , Aldeído Pirúvico/farmacologiaRESUMO
Glycation and the accumulation of advanced glycation end products (AGEs) are known to occur during normal aging but also in the progression of several diseases, such as diabetes. Diabetes type II and aging both lead to impaired wound healing. It has been demonstrated that macrophages play an important role in impaired wound healing, however, the underlying causes remain unknown. Elevated blood glucose levels as well as elevated methylglyoxal (MGO) levels in diabetic patients result in glycation and increase of AGEs. We used MGO to investigate the influence of glycation and AGEs on macrophages. We could show that glycation, but not treatment with AGE-modified serum proteins, increased expression of pro-inflammatory cytokines interleukin 1ß (IL-1ß) and IL-8 but also affected IL-10 and TNF-α expression, resulting in increased inflammation. At the same time, glycation reduced phagocytic efficiency and led to impaired clearance rates of invading microbes and cellular debris. Our data suggest that glycation contributes to changes of macrophage activity and cytokine expression and therefore could support the understanding of disturbed wound healing during aging and diabetes.
Assuntos
Citocinas/metabolismo , Produtos Finais de Glicação Avançada/química , Macrófagos/metabolismo , Fagócitos/metabolismo , Aldeído Pirúvico/química , Envelhecimento/imunologia , Diabetes Mellitus Tipo 2/imunologia , Glicosilação , Humanos , Inflamação/imunologia , Interleucina-10/metabolismo , Interleucina-1beta/metabolismo , Interleucina-8/metabolismo , Ativação de Macrófagos , Espécies Reativas de Oxigênio/metabolismo , Células THP-1 , Fator de Necrose Tumoral alfa/metabolismo , Cicatrização/imunologiaRESUMO
Ascorbic acid better known as vitamin C, is a reducing carbohydrate needed for a variety of functions in the human body. The most important characteristic of ascorbic acid is the ability to donate two electrons, predestining it as a major player in balancing the physiological redox state and as a necessary cofactor in multiple enzymatic hydroxylation processes. Ascorbic acid can be reversibly oxidized in two steps, leading to semidehydroascorbic acid and dehydroascorbic acid, respectively. Further degradation is irreversible and generates highly reactive carbonyl-intermediates. These intermediates are able to induce glycation of proteins, a non-enzymatic and unspecific reaction of carbonyls with amino groups involved to several age-related diseases. In this study, we investigated the effect of ascorbic acid- and dehydroascorbic acid-induced glycation on PC12 cells, which represent a model for neuronal plasticity. We found that both applications of ascorbic acid or dehydroascorbic acid leads to glycation of cellular proteins, but that ascorbic acid interferes more with viability and neurite outgrowth compared with dehydroascorbic acid.