Stress induced aging in mouse eye.

Aging, a universal process that affects all cells in an organism, is a major risk factor for a group of neuropathies called glaucoma, where elevated intraocular pressure is one of the known stresses affecting the tissue. Our understanding of molecular impact of aging on response to stress in retina is very limited; therefore, we developed a new mouse model to approach this question experimentally. Here we show that susceptibility to response to stress increases with age and is primed on chromatin level. We demonstrate that ocular hypertension activates a stress response that is similar to natural aging and involves activation of inflammation and senescence. We show that multiple instances of pressure elevation cause aging of young retina as measured on transcriptional and DNA methylation level and are accompanied by local histone modification changes. Our data show that repeated stress accelerates appearance of aging features in tissues and suggest chromatin modifications as the key molecular components of aging. Lastly, our work further emphasizes the importance of early diagnosis and prevention as well as age-specific management of age-related diseases, including glaucoma.

Early removal of senescent cells protects retinal ganglion cells loss in experimental ocular hypertension.

Experimental ocular hypertension induces senescence of retinal ganglion cells (RGCs) that mimics events occurring in human glaucoma. Senescence-related chromatin remodeling leads to profound transcriptional changes including the upregulation of a subset of genes that encode multiple proteins collectively referred to as the senescence-associated secretory phenotype (SASP). Emerging evidence suggests that the presence of these proinflammatory and matrix-degrading molecules has deleterious effects in a variety of tissues. In the current study, we demonstrated in a transgenic mouse model that early removal of senescent cells induced upon elevated intraocular pressure (IOP) protects unaffected RGCs from senescence and apoptosis. Visual evoked potential (VEP) analysis demonstrated that remaining RGCs are functional and that the treatment protected visual functions. Finally, removal of endogenous senescent retinal cells after IOP elevation by a treatment with senolytic drug dasatinib prevented loss of retinal functions and cellular structure. Senolytic drugs may have the potential to mitigate the deleterious impact of elevated IOP on RGC survival in glaucoma and other optic neuropathies.

Phytochemical analysis and in vitro anti-proliferative activity of Viscum album ethanolic extracts.

BACKGROUND: Viscum album L. (Santalaceae), commonly known as mistletoe, is a hemiparasitic plant traditionally used in complementary cancer treatment. Its antitumor potential is mostly attributed to the presence of aqueous soluble metabolites; however, the use of ethanol as solvent also permits the extraction of pharmacological compounds with antitumor potential. The clinical efficacy of mistletoe therapy inspired the present work, which focuses on ethanolic extracts (V. album "mother tinctures", MT) prepared from different host trees. METHODS: Samples from three European subspecies (album, austriacum, and abietis) were harvested, and five different V. album-MT strains were prepared. The following phytochemical analyses were performed: thin layer chromatography (TLC), high-performance liquid chromatography (HPLC) and liquid chromatography-high resolution mass spectrometry (LC-HRMS). The proliferation assay was performed with WST-1 after incubation of tumor (Yoshida and Molt-4) and fibroblast cell lines (NIH/3 T3) with different MT concentrations (0.5 to 0.05% v/v). The cell death mechanism was investigated by flow cytometry (FACS) using Annexin V-7AAD. RESULTS: Chemical analyses of MT showed the presence of phenolic acids, flavonoids and lignans. The MT flavonoid and viscotoxin contents (mg/g fresh weight) were highest in Quercus robur (9.67 ± 0.85 mg/g) and Malus domestica (3.95 ± 0.58 mg/mg), respectively. The viscotoxin isoform proportions (% total) were also different among the VA subspecies with a higher content of A3 in V. album growing on Abies alba (60.57 ± 2.13). The phytochemical compounds as well as the viscotoxin contents are probably related to the antitumor effects of MT. The cell death mechanisms evaluated by colorimetric and FACS methodologies involved necrotic damage, which was host tree-, time- and dose- dependent, with different selectivity to tumor cells. Mother tincture from V. album ssp. abietis was the most effective at inducing in vitro cellular effects, even when incubated at the smallest concentration tested, probably because of the higher content of VT A3. CONCLUSION: Our results indicate the promising antitumor potential of Viscum album ethanolic extracts and the importance of botanical and phytochemical characterization for in vitro anti-proliferative effects.

Comparison of the King's and MiToS staging systems for ALS.

OBJECTIVE: To investigate and compare two ALS staging systems, King's clinical staging and Milano-Torino (MiToS) functional staging, using data from the LiCALS phase III clinical trial (EudraCT 2008-006891-31). METHODS: Disease stage was derived retrospectively for each system from the ALS Functional Rating Scale-Revised subscores using standard methods. The two staging methods were then compared for timing of stages using box plots, correspondence using chi-square tests, agreement using a linearly weighted kappa coefficient and concordance using Spearman's rank correlation. RESULTS: For both systems, progressively higher stages occurred at progressively later proportions of the disease course, but the distribution differed between the two methods. King's stage 3 corresponded to MiToS stage 1 most frequently, with earlier King's stages 1 and 2 largely corresponding to MiToS stage 0 or 1. The Spearman correlation was 0.54. There was fair agreement between the two systems with kappa coefficient of 0.21. CONCLUSION: The distribution of timings shows that the two systems are complementary, with King's staging showing greatest resolution in early to mid-disease corresponding to clinical or disease burden, and MiToS staging having higher resolution for late disease, corresponding to functional involvement. We therefore propose using both staging systems when describing ALS.

Enhancing the enantioselectivity of an epoxide hydrolase by directed evolution.

[reaction: see text] The epoxide hydrolase (EH) from Aspergillus niger, which shows a selectivity factor of only E = 4.6 in the hydrolytic kinetic resolution of glycidyl phenyl ether, has been subjected to directed evolution for the purpose of enhancing enantioselectivity. After only one round of error-prone polymerase chain reaction (epPCR), enantioselectivity was more than doubled (E = 10.8). The improved mutant enzyme contains three amino acid exchanges, two of which are spatially far from the catalytically active center.

Análise dos efeitos do Método TheraSuit na função motora de uma criança com paralisia cerebral: estudo de caso / Analysis of effects of the TheraSuit Method on the motor development of a child with cerebral palsy a case study

O objetivo deste estudo foi analisar os efeitos do Método TheraSuit® na função motora de uma criança com Paralisia Cerebral (PC). Novos métodos para tratar as disfunções neurológicas e sensoriais causadas pela PC vêm surgindo e despertando interesses nos profissionais da área de saúde. O Método TheraSuit® consiste em um programa individualizado e intensivo de fisioterapia com protocolo de intervenção de 4 semanas, realizado 5 dias por semana por 3 horas diárias. Foram analisados nesse estudo quatro protocolos do Método TheraSuit®, aplicados em intervalos de quatro meses. Em cada um dos quatro protocolos foi utilizada a Medida da Função Motora Grossa (GMFM-66) na avaliação inicial e final. Foi observado na pontuação do GMFM-66 um resultado satisfatório com um score pré protocolos de 38 e pós-protocolos de 45, sem declínio no intervalo entre os protocolos. Conclui-se que com a aplicação do MétodoTheraSuit® houve aprendizado motor observado pelos escores totais do GMFM, monstrando efeitos positivos na função motora da criança do presente estudo

The purpose of this study was to analyze the effects of the TheraSuit Method® on the motor function of a child with Cerebral Palsy (CP). New methods for treating neurological and sensory dysfunctions caused by PC have been emerging and arousing interests in health professionals. The TheraSuit Method® consists of an individualized and intensive physiotherapy program with a 4-week intervention protocol, performed 5 days a week for 3 hours daily. Four protocols of the TheraSuit Method®, applied at four-month intervals, were analyzed in this study. In each of the four protocols, the Thick Motor Function Measure (GMFM-66) was used in the initial and final evaluation. It was observed in the GMFM-66 score a satisfactory score with a before-protocol score of 38 and post-protocols of 45, with no decline in the interval between protocols. It is concluded that with the application of the TheraSuit Method® there was motor learning observed by the total GMFM scores, showing positive effects on the motor function of the child of the present study

Adverse events during longterm low-dose glucocorticoid treatment of polymyalgia rheumatica: a retrospective study.

OBJECTIVE: To assess the occurrence of adverse events in a cohort of patients with polymyalgia rheumatica (PMR), treated with low-dose glucocorticoids (GC). METHODS: This was a retrospective study by review of medical records. RESULTS: We identified 222 patients who had a mean duration of followup of 60 ± 22 months and a mean duration of GC therapy of 46 ± 22 months. We found that 95 patients (43%) had at least 1 adverse event after a mean duration of GC therapy of 31 ± 22 months and a mean cumulative dose of 3.4 ± 2.4 g. In particular, 55 developed osteoporosis, 31 had fragility fractures; 27 developed arterial hypertension; 11 diabetes mellitus; 9 acute myocardial infarction; 3 stroke; and 2 peripheral arterial disease. Univariate analysis showed that the duration of GC treatment was significantly associated with osteoporosis (p < 0.0001), fragility fractures (p < 0.0001), arterial hypertension (p < 0.005), and acute myocardial infarction (p < 0.05). Cumulative GC dose was significantly associated with osteoporosis (p < 0.0001), fragility fractures (p < 0.0001), and arterial hypertension (p < 0.01). The adverse events occurred more frequently after 2 years of treatment. Multivariate analysis showed that GC duration was significantly associated with osteoporosis (adjusted OR 1.02, 95% CI 1.02-1.05) and arterial hypertension (adjusted OR 1.03, 95% CI 1.01-1.06); GC cumulative dose was significantly associated with fragility fractures (adjusted OR 1.4, 95% CI 1.03-1.8). CONCLUSION: Longterm, low-dose GC treatment of PMR is associated with serious adverse events such as osteoporosis, fractures, and arterial hypertension; these adverse events occur mostly after 2 years of treatment.

Biologically active natural products from Mongolian medicinal plants Scorzonera divaricata and Scorzonera pseudodivaricata.

Chromatographic separation of a crude extract obtained from the aerial parts of the Mongolian medicinal plant Scorzonera divaricata yielded the two new quinic acid derivatives feruloylpodospermic acids A and B. Both compounds feature a feruloyl group and two dihydrocaffeoyl substituents. For feruloylpodospermic acid A, the dihydrocaffeic acid substituents were found esterified at positions 1 and 5 of the quinic acid moiety, while the feruloyl group was attached at position 3. For feruloylpodospermic acid B, the substituents were linked at positions 1, 3, and 4. The aerial parts of S. pseudodivaricata that are likewise used in Mongolian traditional medicine yielded two further new natural products, for which the names scorzoneric acid and scorzonerin are proposed. Scorzoneric acid is an unusual phenolic compound featuring a central tetrasubstituted phenyl ring to which a glucose unit is bound, which in turn is substituted by an esterified acyl side chain. Further substituents of the central phenyl ring system include a butan-2-one group, which is linked to a second para-substituted phenyl ring system. Scorzonerin is a matricarin-based sesquiterpene lactone that carries an esterified dihydrocoumaric acid moiety, which in turn is glycosidically bound to glucose. The structures of all new compounds were unambiguously established from NMR (1H, 13C, COSY, HMBC) spectroscopic and mass spectrometric data. The new quinic acid derivatives feruloylpodospermic acids A and B exhibited strong antioxidative activity when analyzed in the DPPH (2,2-diphenyl-1-picrylhydrazyl) assay.

Adulto jovem com paralisia cerebral / Young adult with cerebral palsy

Este estudo considerou os aspectos envolvidos no adulto com paralisia cerebral em relação aos vários sistemas do corpo e analisou o aspecto motor e a presença de dor. Também enfocou as alternativas para o adulto jovem para a continuidade de habilitação, prevenção ou minimização dos efeitos secundários da paralisia cerebral e da idade avançada. Para tanto, foram avaliados nove jovens adultos (18 a 28 anos) e um adulto (51 anos) durante período de dois anos, utilizando-se a Medição da Função Motora Grossa (GMFM). Durante esse período os indivíduos participaram de terapias individuais baseadas no Conceito Bobath, e de terapias em grupo também baseadas no Conceito Bobath, mas associado com aspectos da Educação Condutiva (Método Petõ). Noventa por cento apresentaram ganhos no resultado final do GMFM que variaram de 1,5% a 15,1 %, com média de ganho de 6,2%. Os resultados, portanto, demonstraram que a continuidade de atividade dirigida a mobilidade, controle de postura e dos movimentos integradas a atividades funcionais permitem que o processo de deterioração seja contido ou minimizado para adultos jovens com paralisia cerebral.

This study has considered aspects of several body systems in adults with cerebral palsy emphasizing motor and pain conditions. Options for keeping on the habilitation process, preventing from deterioration, and minimizing secondary effects of cerebral palsy in young adults were also discussed. For this purpose, nine young adult (18 to 28 years of age) and one adult (51 years of age) were assessed during two years by the aplication of the Gross Motor Function Measure (GMFM). During these two years the individuals participated in individual therapies based on Bobath approach as well as in therapeutic group activities also based on Bobath approach associated with some aspects of Conductive Education (Petõ Method). Improvements in GMFM total results varying from 1.5% to 15.1 % (mean of 6.2%) were observed in 90% of the patients. The results have shown, therefore, that the continuity of directed activity aimimg at motor control integrated with functional activities can constrict or minimize the deterioration process in young adults with cerebral palsy.