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111.
Giulio Poli Ivana Barravecchia Gian Carlo Demontis Andrea Sodi Alessandro Saba Stanislao Rizzo Marco Macchia Tiziano Tuccinardi 《Journal of enzyme inhibition and medicinal chemistry》2022,37(1):1765
The human retinal pigment epithelium-specific 65-kDa protein (hRPE65) plays a crucial role within the retinoid visual cycle and several mutations affecting either its expression level or its enzymatic function are associated with inherited retinal diseases such as Retinitis Pigmentosa. The gene therapy product voretigene neparvovec (Luxturna) has been recently approved for treating hereditary retinal dystrophies; however, the treatment is currently accessible only to patients presenting confirmed biallelic mutations that severely impair hRPE65 function, and many reported hRPE65 missense mutations lack sufficient evidences for proving their pathogenicity. In this context, we developed a computational approach aimed at evaluating the potential pathogenic effect of hRPE65 missense variants located on the dimerisation domain of the protein. The protocol evaluates how mutations may affect folding and conformation stability of this protein region, potentially helping clinicians to evaluate the eligibility for gene therapy of patients diagnosed with this type of hRPE65 variant of uncertain significance. 相似文献
112.
Brun P Castagliuolo I Di Leo V Buda A Pinzani M Palù G Martines D 《American journal of physiology. Gastrointestinal and liver physiology》2007,292(2):G518-G525
A small percentage of pathologically obese subjects with fatty livers develop histological signs of necroinflammation and fibrosis, suggesting a variety of cofactors in the pathogenesis of obesity-related liver diseases including nonalcoholic steatohepatitis. Since several observations have linked bacterial endotoxins to liver damage, the aim of this study was to determine the effect of obesity on intestinal mucosal integrity and portal blood endotoxemia in two strains of obese mice: leptin-deficient (ob/ob) and hyperleptinemic (db/db) mice. Murine intestinal mucosal barrier function was assessed using a Ussing chamber, whereas ileum tight junction proteins were analyzed by immunocytochemistry and Western blot analysis. Circulating proinflammatory cytokines and portal blood endotoxin levels were measured by ELISA and the limulus test, respectively. The inflammatory and fibrogenic phenotype of murine hepatic stellate cells (HSCs) was determined by ELISA and quantitative RT-PCR. Ob/ob and db/db mice showed lower intestinal resistance, profoundly modified distribution of occludin and zonula occludens-1 in the intestinal mucosa, and higher circulating levels of inflammatory cytokines and portal endotoxemia compared with lean control mice. Moreover, HSCs isolated from ob/ob and db/db mice showed higher membrane CD14 mRNA levels and more pronounced lipopolysaccharide-induced proinflammatory and fibrogenic responses than HSCs from lean animals. In conclusion, genetically obese mice display enhanced intestinal permeability leading to increased portal endotoxemia that makes HSCs more sensitive to bacterial endotoxins. We suggest that in metabolic syndrome, patients may likewise have a greater intestinal mucosa permeability and increased lipopolysaccharide levels in portal blood that can contribute to the liver inflammatory damage. 相似文献
113.
114.
Rapid and sensitive NMR method for osmolyte determination 总被引:4,自引:0,他引:4
We propose a rapid and sensitive method for osmolyte determination, based on one-dimensional and two-dimensional 1H NMR spectroscopy applied directly on culture of haloalkalophilic Halomonas pantelleriensis and acidothermophilic archaeon Sulfolobus solfataricus, without any extraction procedure. The osmoprotectants hydroxyectoine, ectoine, glutamate, glycine-betaine and treahalose can easily be quantified by integrating the peak areas with respect to an internal standard, and the concentrations evaluated with this method are in excellent agreement with the values previously reported. Furthermore, trace amount of osmoprotectants, often undetectable after extraction procedures, can also be evaluated. 相似文献
115.
Soil seed bank and the effect of needle litter layer on seedling emergence in a tropical pine plantation 总被引:1,自引:0,他引:1
The soil seed bank is the basis for community establishment and permanence and plays a primary role in natural restoration of degraded or altered ecosystems. As part of a restoration project, this study aimed to quantify the soil seed bank and to evaluate the effect of the needle litter layer on seedling emergence. Soil samples from a pine plantation were collected at random in the field and set to germinate in a greenhouse. Half of them were covered by a 6cm layer of dead pine needles simulating field conditions. In the field, 20 x 20cm plots were established, half were left intact and half were cleaned from the litter needles. All four treatments had 15 replicates and seedling emergence was recorded during six months. Soil seed bank density was 1 222/m2 from 17 morphotypes. In the field, the number of morphotypes and seedlings was only 9% and 6% respectively, of those emerged in the greenhouse, possibly due to watering and lack of predation in the latter. In both cases, herbs and graminoids were the dominant emerging seedlings, making up to 70-90% of the total. The needle layer didn't prevent seeds from reaching the soil but strongly reduced (> 50%) seedling emergence, although high variability within treatments resulted in no statistically significant differences. These results show that the needle layer hinders germination and/or emergence of seedlings from the seed bank. Its removal may be a recommended technique to accelerate natural restoration in pine plantations. 相似文献
116.
The cell nucleus is a membrane-surrounded organelle that contains numerous compartments in addition to chromatin. Compartmentalization of the nucleus is now accepted as an important feature for the organization of nuclear processes and for gene expression. Recent studies on nuclear organization of splicing factors in plant cells provide insights into the compartmentalization of the plant cell nuclei and conservation of nuclear compartments between plants and metazoans. 相似文献
117.
Ruth Greussing Hermann Unterluggauer Rafal Koziel Andrea B. Maier Pidder Jansen-Dürr 《Journal of visualized experiments : JoVE》2012,(69)
Proteasome is the main intracellular organelle involved in the proteolytic degradation of abnormal, misfolded, damaged or oxidized proteins 1, 2. Maintenance of proteasome activity was implicated in many key cellular processes, like cell''s stress response 3, cell cycle regulation and cellular differentiation 4 or in immune system response 5. The dysfunction of the ubiquitin-proteasome system has been related to the development of tumors and neurodegenerative diseases 4, 6. Additionally, a decrease in proteasome activity was found as a feature of cellular senescence and organismal aging 7, 8, 9, 10. Here, we present a method to measure ubiquitin-proteasome activity in living cells using a GFP-dgn fusion protein. To be able to monitor ubiquitin-proteasome activity in living primary cells, complementary DNA constructs coding for a green fluorescent protein (GFP)–dgn fusion protein (GFP–dgn, unstable) and a variant carrying a frameshift mutation (GFP–dgnFS, stable 11) are inserted in lentiviral expression vectors. We prefer this technique over traditional transfection techniques because it guarantees a very high transfection efficiency independent of the cell type or the age of the donor. The difference between fluorescence displayed by the GFP–dgnFS (stable) protein and the destabilized protein (GFP-dgn) in the absence or presence of proteasome inhibitor can be used to estimate ubiquitin-proteasome activity in each particular cell strain. These differences can be monitored by epifluorescence microscopy or can be measured by flow cytometry. 相似文献
118.
119.
Biological availability and humic properties of dissolved organic carbon in Lake Balaton (Hungary) 总被引:1,自引:0,他引:1
The nursery role of the Mondego estuary for marine fish species was studied between June 2003 and May 2004. The spatial and
temporal distribution and abundance patterns of 0-group Dicentrarchus
labrax (Linnaeus, 1758), Platichthys
flesus (Linnaeus, 1758) and Solea solea (Linnaeus, 1758) were analyzed based on monthly sampling surveys in five stations along the estuarine gradient. Fishing took
place during the night at low water of spring tides, using a 2 m beam trawl. The spatial patterns of estuarine colonization
were different according to species. D. labrax showed a wider distribution, but the main nursery ground was the same as for S. solea. Highest densities of S. solea juveniles were found in oligohaline areas, with muddy bottoms and high benthic invertebrates availability, while P. flesus occurred mainly in the sandy uppermost areas. D. labrax was found in both these areas. Fish abundance in the estuary mainly reflected seasonal changes. 相似文献
120.