RESUMO
AIMS: The purpose of this study is to explore whether antioxidant DJ-1 protein affects the atrophy of skeletal muscle cell induced by undernutrition. MAIN METHODS: To determine cell atrophic responses, L6 cell line and skeletal primary cells from mouse hind limbs were cultivated under condition of FBS-free and low glucose. Changes of protein expression were analyzed using Western blot. Overexpression and knockdown of DJ-1 was performed in cells to assess its influence on cell atrophic responses. KEY FINDINGS: Undernutrition decreased cell size and increased the abundance of oxidized form and total form of DJ-1 protein in L6 myoblasts. The undernourished cells revealed an elevation in the expression of muscle-specific RING finger-1 (MuRF-1) and atrogin-1, and in the phosphorylations of p38 mitogen-activated protein kinase (MAPK) and stress-activated protein kinase/c-Jun N-terminal kinase compared with control groups. Moreover, DJ-1-knockout mice showed a decrease in cell size and an enhancement in the expression of MuRF-1 and atrogin-1, as well as in the phosphorylation of MAPKs in gastrocnemius muscles; these changes were also observed in L6 cells transfected with siRNA of DJ-1. On the other hand, L6 cells overexpressing full-length DJ-1 did not exhibit the alterations in cell size and ubiquitin ligases seen after undernourished states of control cells. Myotubes differentiated from L6 cells also showed elevated expression of MuRF-1 and atrogin-1 in response to undernutrition. SIGNIFICANCE: These results suggest that DJ-1 protein may contribute to undernutrition-induced atrophy via MAPKs/ubiquitin ligase pathway in skeletal muscle cells.
Assuntos
Sistema de Sinalização das MAP Quinases/fisiologia , Desnutrição/metabolismo , Mioblastos/enzimologia , Proteínas Oncogênicas/deficiência , Peroxirredoxinas/deficiência , Ubiquitina-Proteína Ligases/antagonistas & inibidores , Ubiquitina-Proteína Ligases/metabolismo , Animais , Atrofia/enzimologia , Atrofia/prevenção & controle , Linhagem Celular , Feminino , Masculino , Desnutrição/prevenção & controle , Camundongos , Camundongos Knockout , Mioblastos/patologia , Técnicas de Cultura de Órgãos , Proteína Desglicase DJ-1 , RatosRESUMO
OBJECTIVES: Matrix metalloproteinases (MMPs) are involved in both maintenance of healthy mucosa and mediation of several pathologies. Recently, MMPs and their inhibitors have attracted attention as potential mediators of mucositis. We investigated tissue expression of MMP-3 and MMP-9 over time in a pre-clinical model of irinotecan-induced oral mucositis (OM). MATERIALS AND METHODS: Eighty-one female Dark Agouti rats received either a single dose of irinotecan (200 mg/kg) or vehicle control. Rats were killed at different time points over a 72-h period and tongue mucosa examined histologically. Tissue expression of MMP-3 and MMP-9 was characterized by standard qualitative immunohistochemistry. RESULTS AND DISCUSSION: Epithelial thickness was reduced without any ulceration in the oral mucosa early after chemotherapy. Epithelial atrophy was associated with significant (P < 0.05) upregulation of MMP-3 and MMP-9 in all layers of the oral epithelium. The increase of MMP-3 was also significant (P < 0.05) in lamina propria and submucosa. Most of changes in expression occurred early (1-6 h), coinciding with previously described upregulation of transcription factors and pro-inflammatory cytokines in OM. Tissue expression of MMP-3 and MMP-9 followed different patterns of change over time, suggesting involvement in various aspects of OM pathophysiology. CONCLUSIONS: These findings suggest vital roles played by MMP-3 and MMP-9 during OM pathophysiology. Further research is required to investigate the role of other MMPs and the naturally existing tissue inhibitors of MMPs. Research should also be directed to investigate beneficial effects of MMPs intervention therapies to prevent or reduce the severity of OM.
Assuntos
Camptotecina/análogos & derivados , Metaloproteinase 3 da Matriz/biossíntese , Metaloproteinase 9 da Matriz/biossíntese , Estomatite/induzido quimicamente , Estomatite/enzimologia , Análise de Variância , Animais , Atrofia/induzido quimicamente , Atrofia/enzimologia , Camptotecina/toxicidade , Modelos Animais de Doenças , Feminino , Imuno-Histoquímica , Irinotecano , Mucosa Bucal/efeitos dos fármacos , Mucosa Bucal/enzimologia , Mucosa Bucal/patologia , Distribuição Aleatória , Ratos , Estomatite/metabolismo , Estomatite/patologia , Língua/efeitos dos fármacos , Língua/enzimologia , Língua/patologiaRESUMO
Inhibition of Diacylglycerol O-acyltransferase 1 (DGAT1) has been a mechanism of interest for metabolic disorders. DGAT1 inhibition has been shown to be a key regulator in an array of metabolic pathways; however, based on the DGAT1 KO mouse phenotype the anticipation is that pharmacological inhibition of DGAT1 could potentially lead to skin related adverse effects. One of the aims in developing small molecule DGAT1 inhibitors that target key metabolic tissues is to avoid activity on skin-localized DGAT1 enzyme. In this report we describe a modeling-based approach to identify molecules with physical properties leading to differential exposure distribution. In addition, we demonstrate histological and RNA based biomarker approaches that can detect sebaceous gland atrophy pre-clinically that could be used as potential biomarkers in a clinical setting.
Assuntos
Diacilglicerol O-Aciltransferase/antagonistas & inibidores , Descoberta de Drogas , Inibidores Enzimáticos/efeitos adversos , Inibidores Enzimáticos/farmacologia , Glândulas Sebáceas/efeitos dos fármacos , Glândulas Sebáceas/patologia , Animais , Atrofia/induzido quimicamente , Atrofia/enzimologia , Biomarcadores/metabolismo , Inibidores Enzimáticos/química , Inibidores Enzimáticos/metabolismo , Interações Hidrofóbicas e Hidrofílicas , Masculino , Camundongos , Pele/efeitos dos fármacos , Pele/enzimologia , Pele/metabolismo , Bibliotecas de Moléculas Pequenas/efeitos adversos , Bibliotecas de Moléculas Pequenas/química , Bibliotecas de Moléculas Pequenas/metabolismo , Bibliotecas de Moléculas Pequenas/farmacologiaRESUMO
We analyzed four families that presented with a similar condition characterized by congenital microcephaly, intellectual disability, progressive cerebral atrophy, and intractable seizures. We show that recessive mutations in the ASNS gene are responsible for this syndrome. Two of the identified missense mutations dramatically reduce ASNS protein abundance, suggesting that the mutations cause loss of function. Hypomorphic Asns mutant mice have structural brain abnormalities, including enlarged ventricles and reduced cortical thickness, and show deficits in learning and memory mimicking aspects of the patient phenotype. ASNS encodes asparagine synthetase, which catalyzes the synthesis of asparagine from glutamine and aspartate. The neurological impairment resulting from ASNS deficiency may be explained by asparagine depletion in the brain or by accumulation of aspartate/glutamate leading to enhanced excitability and neuronal damage. Our study thus indicates that asparagine synthesis is essential for the development and function of the brain but not for that of other organs.
Assuntos
Aspartato-Amônia Ligase/deficiência , Aspartato-Amônia Ligase/genética , Encéfalo/enzimologia , Encéfalo/patologia , Predisposição Genética para Doença/genética , Microcefalia/enzimologia , Microcefalia/genética , Adolescente , Animais , Atrofia/complicações , Atrofia/enzimologia , Atrofia/genética , Criança , Feminino , Humanos , Lactente , Recém-Nascido , Deficiência Intelectual/complicações , Deficiência Intelectual/enzimologia , Deficiência Intelectual/genética , Deficiência Intelectual/patologia , Masculino , Camundongos , Camundongos Transgênicos , Microcefalia/complicações , Microcefalia/patologia , Mutação de Sentido Incorreto/genética , Linhagem , SíndromeRESUMO
The increasing popularity of the Cre/loxP recombination system has led to the generation of numerous transgenic mouse lines in which Cre recombinase is expressed under the control of organ- or cell-specific promoters. Alterations in retinal pigment epithelium (RPE), a multifunctional cell monolayer that separates the retinal photoreceptors from the choroid, are prevalent in the pathogenesis of a number of ocular disorders, including age-related macular degeneration. To date, six transgenic mouse lines have been developed that target Cre to the RPE under the control of various gene promoters. However, multiple lines of evidence indicate that high levels of Cre expression can be toxic to mammalian cells. In this study, we report that in the Trp1-Cre mouse, a commonly used transgenic Cre strain for RPE gene function studies, Cre recombinase expression alone leads to RPE dysfunction and concomitant disorganization of RPE layer morphology, large areas of RPE atrophy, retinal photoreceptor dysfunction, and microglial cell activation in the affected areas. The phenotype described herein is similar to previously published reports of conditional gene knockouts that used the Trp1-Cre mouse, suggesting that Cre toxicity alone could account for some of the reported phenotypes and highlighting the importance of the inclusion of Cre-expressing mice as controls in conditional gene targeting studies.
Assuntos
Integrases/fisiologia , Epitélio Pigmentado da Retina/enzimologia , Animais , Atrofia/enzimologia , Atrofia/patologia , Modelos Animais de Doenças , Eletrorretinografia/métodos , Regulação da Expressão Gênica , Integrases/genética , Integrases/metabolismo , Glicoproteínas de Membrana/genética , Glicoproteínas de Membrana/fisiologia , Camundongos , Camundongos Transgênicos , Microglia/patologia , Microglia/fisiologia , Microscopia Eletrônica , Oxirredutases/genética , Oxirredutases/fisiologia , Fenótipo , Células Fotorreceptoras de Vertebrados/fisiologia , Proteínas Recombinantes de Fusão/genética , Distrofias Retinianas/enzimologia , Distrofias Retinianas/patologia , Epitélio Pigmentado da Retina/patologia , Epitélio Pigmentado da Retina/fisiopatologia , Epitélio Pigmentado da Retina/ultraestruturaRESUMO
Degeneration of the cerebrum, cerebellum, and retina in infancy is part of the clinical spectrum of lysosomal storage disorders, mitochondrial respiratory chain defects, carbohydrate glycosylation defects, and infantile neuroaxonal dystrophy. We studied eight individuals from two unrelated families who presented at 2-6 months of age with truncal hypotonia and athetosis, seizure disorder, and ophthalmologic abnormalities. Their course was characterized by failure to acquire developmental milestones and culminated in profound psychomotor retardation and progressive visual loss, including optic nerve and retinal atrophy. Despite their debilitating state, the disease was compatible with survival of up to 18 years. Laboratory investigations were normal, but the oxidation of glutamate by muscle mitochondria was slightly reduced. Serial brain MRI displayed progressive, prominent cerebellar atrophy accompanied by thinning of the corpus callosum, dysmyelination, and frontal and temporal cortical atrophy. Homozygosity mapping followed by whole-exome sequencing disclosed a Ser112Arg mutation in ACO2, encoding mitochondrial aconitase, a component of the Krebs cycle. Specific aconitase activity in the individuals' lymphoblasts was severely reduced. Under restrictive conditions, the mutant human ACO2 failed to complement a yeast ACO1 deletion strain, whereas the wild-type human ACO2 succeeded, indicating that this mutation is pathogenic. Thus, a defect in mitochondrial aconitase is associated with an infantile neurodegenerative disorder affecting mainly the cerebellum and retina. In the absence of noninvasive biomarkers, determination of the ACO2 sequence or of aconitase activity in lymphoblasts are warranted in similarly affected individuals, based on clinical and neuroradiologic grounds.
Assuntos
Aconitato Hidratase/genética , Cerebelo/anormalidades , Mitocôndrias/enzimologia , Mutação , Doenças Neurodegenerativas/genética , Retina/anormalidades , Adolescente , Atrofia/enzimologia , Atrofia/genética , Cerebelo/enzimologia , Criança , Pré-Escolar , Exoma , Éxons , Feminino , Genótipo , Ácido Glutâmico/metabolismo , Heterozigoto , Homozigoto , Humanos , Lactente , Imageamento por Ressonância Magnética/métodos , Masculino , Mitocôndrias/genética , Doenças Neurodegenerativas/enzimologia , Oxirredução , Polimorfismo de Nucleotídeo Único , Retina/enzimologiaRESUMO
High levels of angiotensin-converting-enzyme (ACE) may increase the risk of dementia through blood pressure elevation and subsequent development of cerebral small-vessel disease. However, high ACE levels may also decrease this risk through amyloid degradation which prevents brain atrophy. Within the SMART-MR study, a prospective cohort study among patients with symptomatic atherosclerotic disease, serum ACE levels were measured at baseline and a 1.5 Tesla brain MRI was performed at baseline and after on average (range) 3.9 (3.0-5.8) years of follow-up in 682 persons (mean age 58 ± 10 years). Brain segmentation was used to quantify total, deep, and periventricular white matter lesion (WML) volume, and total brain, cortical gray matter and ventricular volume (%ICV). Lacunar infarcts were rated visually. Regression analyses were used to examine the prospective associations between serum ACE and brain measures. Patients with the highest serum ACE levels (>43.3 U/L) had borderline significantly more progression of deep WML volumes than patients with the lowest ACE levels (<21.8 U/L); mean difference (95% CI) in change was 0.20 (-0.02; 0.43) %ICV. On the contrary, patients with the highest serum ACE levels had significantly less progression of cortical brain atrophy than patients with the lowest ACE levels; mean difference (95% CI) in change was 0.78 (0.21; 1.36) %ICV. Serum ACE was not associated with subcortical atrophy, periventricular WML, or lacunar infarcts. Our results show that higher ACE activity is associated with somewhat more progression of deep WML volume, but with less progression of cortical brain atrophy. This suggests both detrimental and beneficial effects of high ACE levels on the brain.
Assuntos
Encéfalo/enzimologia , Encéfalo/patologia , Fibras Nervosas Mielinizadas/enzimologia , Fibras Nervosas Mielinizadas/patologia , Peptidil Dipeptidase A/sangue , Idoso , Atrofia/sangue , Atrofia/enzimologia , Atrofia/patologia , Biomarcadores/sangue , Infarto Cerebral/sangue , Infarto Cerebral/enzimologia , Infarto Cerebral/patologia , Estudos de Coortes , Progressão da Doença , Feminino , Seguimentos , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Doenças Neurodegenerativas/sangue , Doenças Neurodegenerativas/enzimologia , Doenças Neurodegenerativas/patologia , Estudos ProspectivosRESUMO
Degradation of extracellular matrix chondroitin sulphate proteoglycans (CSPGs) using Chondroitinase ABC (ChABC) is a promising strategy for the treatment of spinal cord injury, with potent effects on promoting functional recovery and anatomical repair in spinal injured animals. We have previously demonstrated that ChABC treatment prevents atrophy of corticospinal projection neurons following spinal injury in adult YFP-H mice. Here, we investigate whether ChABC-mediated repair of the cell body extends to rubrospinal projection neurons (RSNs), whether neuroprotective effects can be sustained long-term and importantly, whether delayed treatment with ChABC can reverse chronic atrophy. Adult YFP-H mice underwent unilateral rubrospinal tract transection and were treated with ChABC or a control enzyme, delivered either acutely post-injury or after a one month delay. Eight weeks following injury and control treatment, RSNs in the injured red nucleus, identified by YFP label and NeuN immunoreactivity, showed severe atrophy, with ~40% loss of mean cell area compared to uninjured neurons in the contralateral red nucleus. Both acute and delayed treatment with ChABC promoted a significant rescue of injured RSNs, restoring cell area to ~80% and ~70%, respectively, of that in uninjured neurons. Thus, we demonstrate for the first time that CSPG degradation in the injured spinal cord not only promotes sustained rescue of cell atrophy when delivered acutely but can also reverse chronic atrophy in descending projection neurons. Thus, modulation of the extracellular matrix can mediate neuroprotective effects both early and late after spinal cord injury.
Assuntos
Condroitina ABC Liase/administração & dosagem , Regeneração Nervosa/fisiologia , Neurônios/enzimologia , Núcleo Rubro/enzimologia , Traumatismos da Medula Espinal/enzimologia , Traumatismos da Medula Espinal/prevenção & controle , Animais , Atrofia/enzimologia , Atrofia/patologia , Atrofia/prevenção & controle , Vértebras Cervicais , Feminino , Masculino , Camundongos , Camundongos Transgênicos , Regeneração Nervosa/efeitos dos fármacos , Neurônios/efeitos dos fármacos , Recuperação de Função Fisiológica/efeitos dos fármacos , Recuperação de Função Fisiológica/fisiologia , Núcleo Rubro/efeitos dos fármacos , Traumatismos da Medula Espinal/patologia , Fatores de Tempo , Resultado do TratamentoRESUMO
OBJECTIVE: Indications for tonsillectomy in recurrent tonsillitis are defined according to the number of episodes of acute bacterial infections in a year. However, little is known about the tonsil immune competence status in patients presenting with recurrent tonsillitis with either hypertrophied or atrophied tonsils, or in patients presenting with obstructive sleep apnoea. In this study we examined the tonsil immune status in children with 3-5 acute recurrent infections a year and in children with obstructive sleep apnoea by comparing the activity of tonsil and adenoid tissue nonspecific alkaline and acid phosphatase. METHODS: Specific activity of tonsil and adenoid tissue nonspecific alkaline and acid phosphatase was investigated in children who underwent tonsillectomy and adenoidectomy for recurrent infection (72 children) and for obstructive sleep apnoea (10 children). Tissue enzyme activities were measured using p-nitrophenylphosphate as a substrate. Tissue samples were examined by the haematoxylin-eosin histological technique. Statistical analyses were performed using SPSS v. 16 software. RESULTS: The tissue nonspecific alkaline phosphatase activity was similar in hypertrophied tonsils in the recurrent infection group and in the obstructive sleep apnoea group (3.437+/-1.226 and 3.978+/-0.762 U/mg of protein, respectively). The enzyme activity in both hypertrophied tonsil groups was significantly higher as compared to atrophied tonsils in the recurrent tonsillitis group, p=0.021 and p=0.006, respectively. The enzyme activity was significantly higher in the adenoids compared to the tonsils from all three groups. Contrary to this, no significant differences were noticed for tonsil and adenoid acid phosphatase activities among the groups. CONCLUSION: Similar acid phosphatase activity in all three groups implies that all three groups have preserved antigen presenting cell activity. In patients with hypertrophied tonsils similar tissue nonspecific alkaline phosphatase activity suggests preserved B cell tonsil immune activity, regardless of the pathology. Patients with atrophied tonsils had significantly lower alkaline phosphatase activity, indicating relative tonsil B cell immune deficiency. Thus, different immunological status in patients presenting with hypertrophied vs. atrophied tonsils could point to a different underlying pathophysiologic mechanism of the disease.
Assuntos
Tonsila Faríngea/enzimologia , Tonsila Faríngea/microbiologia , Fosfatase Alcalina/análise , Fosfatase Alcalina/metabolismo , Tonsila Palatina/enzimologia , Adenoidectomia , Atrofia/enzimologia , Atrofia/imunologia , Atrofia/patologia , Linfócitos B/enzimologia , Linfócitos B/imunologia , Criança , Pré-Escolar , Feminino , Humanos , Concentração de Íons de Hidrogênio , Hipertrofia/enzimologia , Hipertrofia/imunologia , Hipertrofia/patologia , Macrófagos , Masculino , Obstrução Nasal/diagnóstico , Obstrução Nasal/enzimologia , Obstrução Nasal/cirurgia , Nitrofenóis , Tonsila Palatina/imunologia , Tonsila Palatina/patologia , Recidiva , Apneia Obstrutiva do Sono/diagnóstico , Apneia Obstrutiva do Sono/enzimologia , Tonsilectomia , TonsiliteRESUMO
Aging is associated with deficiencies in the prefrontal cortex, including working memory impairment and compromised integrity of neuronal dendrites. Although protein kinase C (PKC) is implicated in structural plasticity, and overactivation of PKC results in working memory impairments in young animals, the role of PKC in prefrontal cortical impairments in the aged has not been examined. This study provides the first evidence that PKC activity is associated with prefrontal cortical dysfunction in aging. Pharmacological inhibition of PKC with chelerythrine rescued working memory impairments in aged rats and enhanced working memory in aged rhesus monkeys. Improvement correlated with age, with older monkeys demonstrating a greater degree of improvement following PKC inhibition. Furthermore, PKC activity within the prefrontal cortex was inversely correlated with the length of basal dendrites of prefrontal cortical neurons, as well as with working memory performance in aged rats. Together these findings indicate that PKC is dysregulated in aged animals and that PKC inhibitors may be useful in the treatment of cognitive deficits in the elderly.
Assuntos
Envelhecimento/metabolismo , Atrofia/enzimologia , Transtornos Cognitivos/enzimologia , Córtex Pré-Frontal/enzimologia , Proteína Quinase C/metabolismo , Envelhecimento/patologia , Envelhecimento/psicologia , Animais , Atrofia/patologia , Atrofia/fisiopatologia , Benzofenantridinas/farmacologia , Transtornos Cognitivos/patologia , Transtornos Cognitivos/fisiopatologia , Dendritos/enzimologia , Dendritos/patologia , Espinhas Dendríticas/enzimologia , Espinhas Dendríticas/patologia , Modelos Animais de Doenças , Inibidores Enzimáticos/farmacologia , Feminino , Macaca mulatta , Masculino , Degeneração Neural/enzimologia , Degeneração Neural/patologia , Neurônios/enzimologia , Neurônios/patologia , Testes Neuropsicológicos , Córtex Pré-Frontal/patologia , Córtex Pré-Frontal/fisiopatologia , Proteína Quinase C/antagonistas & inibidores , Ratos , Ratos Sprague-DawleyRESUMO
Isolated sulfite oxidase deficiency is a rare genetic neurometabolic disease. The first symptoms of this disorder (similar to symptoms of ischemic events) may lead to misdiagnosis and to subsequent birth of affected children in these families. This study characterizes the magnetic resonance (MR) imaging and (for the first time, to our knowledge) the MR spectroscopy features of isolated sulfite oxidase deficiency to provide a means for early and correct diagnosis. Three patients with isolated sulfite oxidase deficiency are studied who manifested intractable seizures and severe hypotonia in the immediate postnatal period with an unknown diagnosis, despite extensive workup. MR imaging and proton MR spectroscopy examinations were performed early in the neonatal period in 2 infants and after 5 months in the third infant. The prominent MR features were early cystic white matter damage, accompanied by profound cerebral atrophy in the third infant. Compared with hypoxic-ischemic disorder, MR findings in isolated sulfite oxidase deficiency demonstrate a more severe condition, without subsequent recovery. The MR spectroscopy studies indicate early onset of energetic and metabolic imbalance. Urine stick findings demonstrated high sulfite levels in 2 patients, and the final diagnosis was subsequently made based on molecular, biochemical, and genetic findings. Magnetic resonance imaging and MR spectroscopy measurements may help differentiate isolated sulfite oxidase deficiency from hypoxic-ischemic condition in patients in whom this diagnosis is not clinically suspected and may lead to further genetic antenatal inquiry that might prevent the birth of other infants affected with this severe and incurable congenital disease.
Assuntos
Encefalopatias Metabólicas Congênitas/patologia , Encéfalo/patologia , Predisposição Genética para Doença/genética , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/deficiência , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/genética , Aminoácidos Sulfúricos/metabolismo , Atrofia/enzimologia , Atrofia/genética , Atrofia/fisiopatologia , Encéfalo/enzimologia , Encéfalo/fisiopatologia , Dano Encefálico Crônico/enzimologia , Dano Encefálico Crônico/genética , Dano Encefálico Crônico/fisiopatologia , Encefalopatias Metabólicas Congênitas/enzimologia , Encefalopatias Metabólicas Congênitas/fisiopatologia , Diagnóstico Diferencial , Diagnóstico Precoce , Epilepsia/enzimologia , Epilepsia/genética , Epilepsia/fisiopatologia , Feminino , Marcadores Genéticos/genética , Humanos , Lactente , Recém-Nascido , Imageamento por Ressonância Magnética , Espectroscopia de Ressonância Magnética , Masculino , Sulfitos/urinaRESUMO
RATIONALE: Unloading the diaphragm via mechanical ventilation (MV) results in rapid diaphragmatic fiber atrophy. It is unknown whether the myonuclear domain (cytoplasmic myofiber volume/myonucleus) of diaphragm myofibers is altered during MV. OBJECTIVE: We tested the hypothesis that MV-induced diaphragmatic atrophy is associated with a loss of myonuclei via a caspase-3-mediated, apoptotic-like mechanism resulting in a constant myonuclear domain. METHODS: To test this postulate, Sprague-Dawley rats were randomly assigned to a control group or to experimental groups exposed to 6 or 12 h of MV with or without administration of a caspase-3 inhibitor. MEASUREMENTS AND MAIN RESULTS: After 12 h of MV, type I and type IIa diaphragm myofiber areas were decreased by 17 and 23%, respectively, and caspase-3 inhibition attenuated this decrease. Diaphragmatic myonuclear content decreased after 12 h of MV and resulted in the maintenance of a constant myonuclear domain in all fiber types. Both 6 and 12 h of MV resulted in caspase-3-dependent increases in apoptotic markers in the diaphragm (e.g., number of terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling positive nuclei and DNA fragmentation). Caspase-3-dependent increases in apoptotic markers occurred after 6 h of MV, before the onset of myofiber atrophy. CONCLUSIONS: Collectively, these data support the hypothesis that the myonuclear domain of diaphragm myofibers is maintained during prolonged MV and that caspase-3-mediated myonuclear apoptosis contributes to this process.
Assuntos
Apoptose , Caspase 3/metabolismo , Diafragma/enzimologia , Diafragma/patologia , Respiração Artificial/efeitos adversos , Animais , Atrofia/enzimologia , Biomarcadores/análise , Biomarcadores/metabolismo , Caspase 3/análise , Feminino , Ratos , Ratos Sprague-DawleyRESUMO
It was reported that 15 of 19 consultation cases of prostatic partial atrophy were a-methylacyl coenzyme A racemase (AMACR)-positive. We investigated partial atrophy cases from a single institution using a standard AMACR immunostaining method. Immunohistochemical analysis was performed using an antibody cocktail containing p63, high-molecular-weight keratin (34bE12), and AMACR antibodies on 122 foci of partial atrophy. AMACR staining was analyzed in partial atrophy (n = 122) and compared with adjacent benign glands (n = 122) and prostatic carcinomas (n = 28). Of 122 foci of partial atrophy, 38 (31.1%) showed AMACR immunoreactivity. Typically, AMACR staining was weak or moderate. In addition, 82 (67.2%) showed patchy to negative distribution of basal cells. Partial atrophy may show AMACR immunoreactivity when using a 34bE12, p63, and AMACR antibody cocktail staining method. Compounding this problem, focal lack of basal cells may be seen. However, the AMACR staining pattern of partial atrophy is usually comparable to that of adjacent benign glands and substantially different from adenocarcinoma.
Assuntos
Próstata/enzimologia , Doenças Prostáticas/enzimologia , Racemases e Epimerases/metabolismo , Adenocarcinoma/enzimologia , Adenocarcinoma/patologia , Adulto , Idoso , Atrofia/enzimologia , Atrofia/patologia , Biomarcadores/metabolismo , Biópsia por Agulha , Humanos , Técnicas Imunoenzimáticas , Masculino , Pessoa de Meia-Idade , Próstata/patologia , Doenças Prostáticas/patologia , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/patologiaAssuntos
Anti-Inflamatórios não Esteroides/uso terapêutico , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/patologia , Idoso , Atrofia/enzimologia , Ciclo-Oxigenase 2/fisiologia , Humanos , Inflamação/enzimologia , Inflamação/patologia , Masculino , Próstata/patologia , Regulação para Cima/fisiologiaRESUMO
Neurologic disease in glutaryl-CoA dehydrogenase (GCDH) deficiency usually presents with acute encephalopathic crises before 2 years of age. The authors report two previously asymptomatic patients with macrocephaly presenting with progressive neurologic deterioration and a severe leukoencephalopathy during adolescence or adulthood.
Assuntos
Encefalopatias Metabólicas Congênitas/enzimologia , Encefalopatias Metabólicas Congênitas/fisiopatologia , Córtex Cerebral/enzimologia , Córtex Cerebral/fisiopatologia , Glutaril-CoA Desidrogenase/deficiência , Idade de Início , Idoso , Atrofia/enzimologia , Atrofia/genética , Atrofia/fisiopatologia , Encefalopatias Metabólicas Congênitas/genética , Carnitina/uso terapêutico , Córtex Cerebral/patologia , Transtornos Cognitivos/enzimologia , Transtornos Cognitivos/genética , Transtornos Cognitivos/fisiopatologia , Análise Mutacional de DNA , Progressão da Doença , Alimentos Formulados , Humanos , Ventrículos Laterais/patologia , Imageamento por Ressonância Magnética , Masculino , Transtornos dos Movimentos/enzimologia , Transtornos dos Movimentos/genética , Transtornos dos Movimentos/fisiopatologia , Mutação/genética , Fibras Nervosas Mielinizadas/patologia , Resultado do TratamentoRESUMO
Despite biological support for a role of angiotensin converting enzyme (ACE) in Alzheimer's disease (AD), studies assessing the ACE I/D polymorphism in AD are conflicting. We re-evaluated this association in the Rotterdam Study, a population-based cohort study. The mechanism of association was further explored by adjusting for vascular factors, and by analysing atrophy, white matter lesions and infarcts on MRI in non-demented individuals. Genotypes were available for 6488 participants. During average follow-up of 6 years 250 subjects developed AD. MRI data were available for 494 non-demented participants. Homozygosity for the I-allele conferred a slightly increased risk of AD compared to carrying a D-allele (RR 1.12 (95% CI 0.99-1.25)). This increase was only significant in women, and independent of vascular factors (RR 1.39 (95% CI 1.14-1.69)). Non-demented women with the II genotype had smaller hippocampal and amygdalar volumes. Vascular pathology was not significantly associated with ACE. This suggests a modest but significant increase in risk of AD and early AD pathology in women homozygous for the ACE I-allele independent of vascular factors.
Assuntos
Doença de Alzheimer/genética , Tonsila do Cerebelo/patologia , Atrofia/genética , Hipocampo/patologia , Peptidil Dipeptidase A/genética , Polimorfismo Genético/genética , Idoso , Doença de Alzheimer/enzimologia , Doença de Alzheimer/patologia , Atrofia/enzimologia , Atrofia/patologia , Transtornos Cerebrovasculares/enzimologia , Transtornos Cerebrovasculares/genética , Transtornos Cerebrovasculares/patologia , Estudos de Coortes , Análise Mutacional de DNA , Feminino , Predisposição Genética para Doença/genética , Testes Genéticos , Genótipo , Homozigoto , Humanos , Imageamento por Ressonância Magnética , Masculino , Pessoa de Meia-Idade , Fibras Nervosas Mielinizadas/patologia , Países Baixos , Fatores SexuaisRESUMO
Small atrophic prostate cancers on needle biopsy are rare and difficult to distinguish from benign atrophy on needle biopsy. We report on a study of 23 needle biopsy specimens with small foci of atrophic prostate cancer from the consult service of one of the authors. In 19 cancer cases the atrophic component was pure; in 4 cases it was dominant with a minor (<5%) nonatrophic cancer component. These atrophic cancers and 16 cases of florid benign atrophy on needle biopsy were examined by immunohistochemistry for alpha-methylacyl-CoA-racemase (AMACR). All cases of cancer and atrophy were verified immunohistochemically with antibodies to basal cells (34betaE12 and p63). AMACR staining were scored as 1+ (5% to 25% of glands expressing AMACR), 2+ (26% to 50% of glands expressing AMACR), or 3+ (>50% of glands expressing AMACR). Positive staining was defined as staining above that of surrounding benign glands. AMACR was expressed in 69.6% of atrophic prostate cancers (3+, 11 cases; 2+, 3 cases; 1+, 2 cases); 30.4% (7 cases) of atrophic prostate cancer exhibited no AMACR expression. In the 4 cases with a few glands of ordinary (nonatrophic) prostate cancer, the nonatrophic cancer demonstrated more intense and a greater extent of AMACR staining. Fourteen cases (87.5%) of benign atrophy showed no AMACR expression. In 2 cases (12.5%) of benign atrophy, background immunostaining made it difficult to assess AMACR expression. We conclude that AMACR immunostaining alone is not sufficiently discriminatory in the differential diagnosis of atrophic prostate cancer versus benign atrophy. Atrophic prostate cancers are not as frequently or as strongly positive as ordinary prostate cancer. Using a panel of immunostains including AMACR, 34betaE12 and p63 (positive AMACR immunostaining along with negative basal cell markers) is recommended in the differentiation of atrophic prostate cancer and benign atrophy.
Assuntos
Adenocarcinoma/diagnóstico , Imuno-Histoquímica , Neoplasias da Próstata/diagnóstico , Adenocarcinoma/enzimologia , Atrofia/enzimologia , Biomarcadores Tumorais/análise , Biópsia por Agulha , Humanos , Masculino , Neoplasias da Próstata/enzimologia , Racemases e EpimerasesRESUMO
Various phospholipases are thought to be associated with the in vitro apoptosis of thymocytes. In the present study, the in vivo phospholipase D (PLD) activity of rat thymus was studied after whole-body X-irradiation or injection of dexamethasone (DEX). Using exogenous [14C]dipalmitoyl phosphatidylcholine (PC) as the substrate, an elevation of oleate-activated PLD activity was observed during thymic atrophy. The activity increases were sevenfold at 48 hr after 5-Gy irradiation and fourfold at 72 hr after injection of 5 mg/kg DEX. The elevation of PLD activity appeared to parallel extensive thymus shrinkage. An increased level of thymic phosphatidic acid (PA), the presumed physiological product of PLD action on PC, was also detected. By comparing the acyl chains of PA with those of other phospholipids, PA appeared to originate from PC. To assess the role of PLD during thymic atrophy, thymocytes and stromal cells were isolated. Although thymocytes themselves exhibited significant PLD activation, the major elevation in PLD activity (greater than fourfold) was found in isolated stromal cells. PLD was also activated during in vitro phagocytosis of apoptotic thymocytes by the macrophage-like cell line P388D1. This in vitro phagocytosis was significantly inhibited by PLD action blockers, such as 2,3-diphosphoglycerate and 1-butanol. These observations strongly suggest that the alteration of oleate-activated PLD activity is part of an in vivo event in the progression of thymic atrophy, including phagocytic clearance of apoptotic thymocytes.
Assuntos
Ácido Oleico/farmacologia , Fosfolipase D/metabolismo , Timo/enzimologia , Animais , Apoptose , Atrofia/enzimologia , Atrofia/imunologia , Cromatografia Líquida de Alta Pressão , Fragmentação do DNA/imunologia , Dexametasona/farmacologia , Feminino , Macrófagos/enzimologia , Fagocitose/imunologia , Ácidos Fosfatídicos/análise , Ratos , Ratos Wistar , Timo/efeitos dos fármacos , Timo/imunologia , Irradiação Corporal TotalRESUMO
The cricopharyngeal muscle (CPM) is essential for normal deglutition. Pharyngeal dysphagia commonly results from impaired or uncoordinated CPM dilation. Dysfunction of the CPM has also been implicated in the genesis of Zenker's (pharyngoesophageal) diverticulum. Despite the CPM's significance, little is understood about its morphology. We studied CPM biopsy specimens from 20 patients with Zenker's diverticulum and from 5 fresh cadaver patients with detailed histologic techniques to include fiber size and shape and adenosine triphosphatase, reduced nicotinamide adenine dinucleotide, trichrome, succinate dehydrogenase, cytochrome C oxidase, periodic acid-Schiff reaction, oil red O, acid phosphatase, Congo red, crystal violet, and monoadenylate deaminase stains. The normal CPM has unique morphological characteristics, with some myofibers having staining properties that are a hybrid between striated muscle and muscle spindle. The variable orientation of the muscle fibers is also different from that of most other striated musculature. Of the 20 Zenker CPM specimens, 4 specimens did not reveal any significant differences from controls (2 of which had insufficient amounts of tissue for complete analysis). In the remaining 16 specimens, several abnormalities existed, including excessive size variation (16/16), grouping of atrophic fibers (9/16), target or targetoid formations (4/16), cores (2/16), and ragged red fibers (2/16). The final pathological pattern of the 16 specimens was neurogenic in 7, myopathic in 4, and mixed (with neurogenic predominance) in the remaining 5. Two specimens contained significant lymphocytic inflammatory infiltrates. We conclude that the unique neuromuscular function of the CPM in deglutition is likely due to its fiber orientation and the hybrid nature of some of the myofibers. Morphological disturbances of the CPM impair its dilation and may account for the development of Zenker's diverticulum. This disturbance is most often due to progressive denervation of the CPM.
Assuntos
Esôfago/patologia , Músculos Faríngeos/patologia , Divertículo de Zenker/patologia , Adenosina Trifosfatases/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Atrofia/enzimologia , Atrofia/patologia , Biópsia , Esôfago/enzimologia , Feminino , Humanos , Nervos Laríngeos/patologia , Masculino , Pessoa de Meia-Idade , Fibras Musculares Esqueléticas/enzimologia , Fibras Musculares Esqueléticas/patologia , NAD/metabolismo , Degeneração Neural/patologia , Músculos Faríngeos/enzimologia , Nervo Vago/patologia , Divertículo de Zenker/enzimologiaRESUMO
Cyclooxygenase-2 (COX-2) is the inducible isoform of the rate-limiting enzymes that convert arachidonic acid to proinflammatory prostaglandins as well as a primary target for nonsteroidal anti-inflammatory drugs. Accumulating evidence suggests that up-regulation of COX-2 is associated with carcinogenesis in multiple organ systems including the large bowel, lung, breast, and prostate. In this report, we examine the expression of COX-2 protein and mRNA in prostate tissue containing various lesions and in prostate cancer cell lines. In the cell lines, LNCaP, DU145, PC-3, and TSU, COX-2 protein expression was undetectable under basal conditions but could be induced transiently by phorbol ester treatment in PC-3 and TSU cells, but not in DU145 and LNCaP cells. Immunohistochemical analysis of 144 human prostate cancer cases suggested that, in contrast to several previous reports, there was no consistent overexpression of COX-2 in established prostate cancer or high-grade prostatic intraepithelial neoplasia, as compared with adjacent normal prostate tissue. Positive staining was seen only in scattered cells (<1%) in both tumor and normal tissue regions but was much more consistently observed in areas of proliferative inflammatory atrophy, lesions that have been implicated in prostatic carcinogenesis. Staining was also seen at times in macrophages. Western blotting and quantitative RT-PCR analyses confirmed these patterns of expression. These results suggest that if nonsteroidal anti-inflammatory drugs are indeed chemopreventive and/or chemotherapeutic for prostate cancer, their effects are likely to be mediated by modulating COX-2 activity in non-PCa cells (either inflammatory cells or atrophic epithelial cells) or by affecting a COX-2-independent pathway.