Fine tuning of metal-specific activity in the Mn-like group of cambialistic superoxide dismutases

RSC Advances ◽  
2015 ◽  
Vol 5 (107) ◽  
pp. 87876-87887 ◽  
Author(s):  
Irene Russo Krauss ◽  
Antonello Merlino ◽  
Andrea Pica ◽  
Rosario Rullo ◽  
Alessandra Bertoni ◽  
...  

Metal-dependent activity and X-ray structures of superoxide dismutase (SOD) fromStreptococcus mutansandStreptococcus thermophilussuggest that they are members of the Mn-like group of cambialistic SODs.

2021 ◽  
Vol 2 (1) ◽  
Author(s):  
Fausta Loffredo ◽  
Loredana Tammaro ◽  
Tiziana Di Luccio ◽  
Carmela Borriello ◽  
Fulvia Villani ◽  
...  

AbstractTungsten disulfide (WS2) nanotubes (NTs) are examined here as a filler for polylactide (PLA) for their ability to accelerate PLA crystallization and for their promising biocompatibility in relevant to biomedical applications of PLA-WS2 nanocomposites. In this work, we have studied the structural and thermal properties of PLA-WS2 nanocomposite films varying the concentration of WS2 NTs from 0 (neat PLA) to 0.6 wt%. The films were uniaxially drawn at 90 °C and annealed at the same temperature for 3 and 10 min. Using wide angle x-ray scattering, Raman spectroscopy and differential scanning calorimetry, we probed the effects of WS2 NT addition on the structure of the PLA films at various stages of processing (unstretched, stretching, annealing). We found that 0.6 wt% of WS2 induces the same level of crystallinity in as stretched PLA-WS2 as annealing in neat PLA for 10 min. These data provide useful insights into the role of WS2 NTs on the structural evolution of PLA-WS2 composites under uniaxial deformation, and extend their applicability to situations where fine tuning of PLA crystallinity is desirable.


2021 ◽  
Vol 29 (1) ◽  
pp. 19-36
Author(s):  
Çağín Polat ◽  
Onur Karaman ◽  
Ceren Karaman ◽  
Güney Korkmaz ◽  
Mehmet Can Balcı ◽  
...  

BACKGROUND: Chest X-ray imaging has been proved as a powerful diagnostic method to detect and diagnose COVID-19 cases due to its easy accessibility, lower cost and rapid imaging time. OBJECTIVE: This study aims to improve efficacy of screening COVID-19 infected patients using chest X-ray images with the help of a developed deep convolutional neural network model (CNN) entitled nCoV-NET. METHODS: To train and to evaluate the performance of the developed model, three datasets were collected from resources of “ChestX-ray14”, “COVID-19 image data collection”, and “Chest X-ray collection from Indiana University,” respectively. Overall, 299 COVID-19 pneumonia cases and 1,522 non-COVID 19 cases are involved in this study. To overcome the probable bias due to the unbalanced cases in two classes of the datasets, ResNet, DenseNet, and VGG architectures were re-trained in the fine-tuning stage of the process to distinguish COVID-19 classes using a transfer learning method. Lastly, the optimized final nCoV-NET model was applied to the testing dataset to verify the performance of the proposed model. RESULTS: Although the performance parameters of all re-trained architectures were determined close to each other, the final nCOV-NET model optimized by using DenseNet-161 architecture in the transfer learning stage exhibits the highest performance for classification of COVID-19 cases with the accuracy of 97.1 %. The Activation Mapping method was used to create activation maps that highlights the crucial areas of the radiograph to improve causality and intelligibility. CONCLUSION: This study demonstrated that the proposed CNN model called nCoV-NET can be utilized for reliably detecting COVID-19 cases using chest X-ray images to accelerate the triaging and save critical time for disease control as well as assisting the radiologist to validate their initial diagnosis.


2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Benoît Bragantini ◽  
Christophe Charron ◽  
Maxime Bourguet ◽  
Arnaud Paul ◽  
Decebal Tiotiu ◽  
...  

AbstractBiogenesis of eukaryotic box C/D small nucleolar ribonucleoproteins initiates co-transcriptionally and requires the action of the assembly machinery including the Hsp90/R2TP complex, the Rsa1p:Hit1p heterodimer and the Bcd1 protein. We present genetic interactions between the Rsa1p-encoding gene and genes involved in chromatin organization including RTT106 that codes for the H3-H4 histone chaperone Rtt106p controlling H3K56ac deposition. We show that Bcd1p binds Rtt106p and controls its transcription-dependent recruitment by reducing its association with RNA polymerase II, modulating H3K56ac levels at gene body. We reveal the 3D structures of the free and Rtt106p-bound forms of Bcd1p using nuclear magnetic resonance and X-ray crystallography. The interaction is also studied by a combination of biophysical and proteomic techniques. Bcd1p interacts with a region that is distinct from the interaction interface between the histone chaperone and histone H3. Our results are evidence for a protein interaction interface for Rtt106p that controls its transcription-associated activity.


1985 ◽  
Vol 40 (5-6) ◽  
pp. 364-372 ◽  
Author(s):  
P. Zipper ◽  
R. Wilfing ◽  
M. Kriechbaum ◽  
H. Durchschlag

Abstract The sulfhydryl enzyme malate synthase from baker’s yeast was X-irradiated with 6 kGy in air-saturated aqueous solution (enzyme concentration: ≃ 10 mg/ml; volume: 120 μl), in the absence or presence of the specific scavengers formate, superoxide dismutase, and catalase. After X-irradiation, a small aliquot of the irradiated solutions was tested for enzymic activity while the main portion was investigated by means of small-angle X-ray scattering. Additionally, an unir­radiated sample without additives was investigated as a reference. Experiments yielded the fol­lowing results: 1. X-irradiation in the absence of the mentioned scavengers caused considerable aggregation, fragmentation, and inactivation of the enzyme. The dose Dt37 for total (= repairable + non­-repayable) inactivation resulted as 4.4 kGy. The mean radius of gyration was found to be about 13 nm. The mean degree of aggregation was obtained as 5.7, without correction for fragmenta­tion. An estimation based on the thickness factor revealed that about 19% of material might be strongly fragmented. When this amount of fragments was accordingly taken into account, a value of 7.1 was obtained as an upper limit for the mean degree of aggregation. The observed retention of the thickness factor and the finding of two different cross-section factors are in full accord with the two-dimensional aggregation model established previously (Zipper and Durchschlag, Radiat. Environ. Biophys. 18, 99 - 121 (1980)). 2. The presence of catalytic amounts of superoxide dismutase and/or catalase, in the absence of formate, during X-irradiation reduced both aggregation and inactivation significantly. 3. The presence of formate (10 or 100 mᴍ) during X-irradiation led to a strong decrease of aggregation and inactivation. This effect was more pronounced with the higher formate concen­tration or when superoxide dismutase and/or catalase were simultaneously present during X-irradiation. The presence of formate also reduced the amount of fragments significantly. 4. The results clearly show that the aggregation and inactivation of malate synthase upon X-irradiation in aqueous solution are mainly caused by OH·; to a minor extent O·̄2 and H2O2 are additionally involved in the damaging processes.


2015 ◽  
Vol 51 (3) ◽  
pp. 584-587 ◽  
Author(s):  
E. Luchinat ◽  
A. Gianoncelli ◽  
T. Mello ◽  
A. Galli ◽  
L. Banci

Combined in-cell NMR spectroscopy, X-ray fluorescence and optical fluorescence microscopies allow describing the intracellular maturation states of human SOD1.


1961 ◽  
Vol 39 (6) ◽  
pp. 1393-1407 ◽  
Author(s):  
Michael Shaw

Wang (Can. J. Botany, 38, 635–642 (1960)) concluded that the accumulation of radioactivity observed on radioautographs at infection sites on rusted leaves fed with C14-labelled substances was 'apparent' rather than real. The ‘accumulation ratio’ is defined as the ratio of the specific activities (c.p.m./mg dry weight of intact tissue) of rust-infected to uninfected areas of infected leaves. Theoretical considerations relating to the radioautography of leaves labelled with C14 and to the measurement of ‘accumulation ratios’ by extraction of C14-labelled substances from rusted and uninfected segments of infected leaves, as well as experimental data, show that Wang's conclusion is not generally applicable.Experimentally, it was shown using polymethacrylate C14 sources that differences in distance between sources and X-ray film of the order of 100 μ had no effect on the intensity of autoradiographs. Rust-infected leaves, fed with radioactive glucose, were radiographed between X-ray plates. Localization of radioactivity at infection sites was observed on both ‘dorsal’ and ‘ventral’ radiographs, indicating a real accumulation per unit area. Ventral were more radioactive than dorsal surfaces. The main development of the fungus occurred on the former. Radioautography revealed that C14 from glucose-1-C14, glucose-6-C14, and uniformly labelled glucose fed to excised wheat leaves became localized at 10-day-old rust infections in 2 hours. ‘Accumulation ratios’ calculated from the specific activity of leaf segments remained close to 1.0 for at least 6 hours after introduction of the tracer, but increased to more than 2 after 24 hours. When ‘accumulation ratios’ were calculated from the specific activities of individual pustules (excised with a punch 1 mm in diameter) and interpustular disks, values greater than 1 were observed in 2 hours, thus confirming the results of autoradiography. Differences between the ‘accumulation ratios’ observed with glucose-6-C14 and glucose-1-C14 were consistent with an increased role of the pentose phosphate pathway at infection sites. Incorporation of C14 from uniformly labelled glucose into the alcohol-insoluble fraction of rusted leaf segments was 2.5-fold that in uninfected segments in 6 hours and 3.65-fold in 24 hours. The humin formed during hydrochloric acid hydrolysis accounted for approximately 50% of the activity of the alcohol-insoluble material. The ‘accumulation ratio’ for the alcohol-soluble material was only 1.56 after 24 hours.All the results support the view (Shaw and Samborski, Can. J. Botany, 34, 389–405 (1956)) that there is a quantitative, metabolically dependent accumulation of C14 from radioactive glucose at vigorous rust infections. The relative roles of fungus and host in this process are discussed briefly.


2010 ◽  
Vol 192 (18) ◽  
pp. 4776-4785 ◽  
Author(s):  
Rabeb Dhouib ◽  
Françoise Laval ◽  
Frédéric Carrière ◽  
Mamadou Daffé ◽  
Stéphane Canaan

ABSTRACT MSMEG_0220 from Mycobacterium smegmatis, the ortholog of the Rv0183 gene from M. tuberculosis, recently identified and characterized as encoding a monoacylglycerol lipase, was cloned and expressed in Escherichia coli. The recombinant protein (rMSMEG_0220), which exhibits 68% amino acid sequence identity with Rv0183, showed the same substrate specificity and similar patterns of pH-dependent activity and stability as the M. tuberculosis enzyme. rMSMEG_0220 was found to hydrolyze long-chain monoacylglycerol with a specific activity of 143 ± 6 U mg−1. Like Rv0183 in M. tuberculosis, MSMEG_0220 was found to be located in the cell wall. To assess the in vivo role of the homologous proteins, an MSMEG_0220 disrupted mutant of M. smegmatis (MsΔ0220) was produced. An intriguing change in the colony morphology and in the cell interaction, which were partly restored in the complemented mutant containing either an active (ComMsΔ0220) or an inactive (ComMsΔ0220S111A) enzyme, was observed. Growth studies performed in media supplemented with monoolein showed that the ability of both MsΔ0220 and ComMsΔ0220S111A to grow in the presence of this lipid was impaired. Moreover, studies of the antimicrobial susceptibility of the MsΔ0220 strain showed that this mutant is more sensitive to rifampin and more resistant to isoniazid than the wild-type strain, pointing to a critical structural role of this enzyme in mycobacterial physiology, in addition to its function in the hydrolysis of exogenous lipids.


1985 ◽  
Vol 249 (5) ◽  
pp. C393-C397 ◽  
Author(s):  
K. Asayama ◽  
R. L. Janco ◽  
I. M. Burr

Radioimmunoassays for both human copper-zinc and manganous superoxide dismutases (Cu-Zn SOD and Mn SOD, respectively) have been developed, validated, and utilized to measure the concentrations of these enzymes in cultured monocytes. Monocyte Mn SOD increased 4.7-fold over basal during 3 days of culture, an increase that was markedly enhanced by stimulation with bacterial lipopolysaccharide (LPS). Cu-Zn SOD showed a transient decrease over the culture period but was unaffected by LPS. Stimulation with muramyl dipeptide had minimal effect on Mn SOD and no effect on Cu-Zn SOD during culture, even at a concentration capable of activating the monocytes, as defined by zymosan-induced superoxide production.


2009 ◽  
Vol 61 (3) ◽  
pp. 441-446 ◽  
Author(s):  
Ana Vuleta ◽  
Branka Tucic

Thermal dependence of the enzymes SOD, CAT, and POD was investigated in leaves of Iris pumila plants inhabiting two contrasting light environments, a sun-exposed dune site and a woodland understory. At the same assay temperature, both the specific activity and the activation energy of SOD and CAT were higher in plants inhabiting vegetation shade than in those experiencing full sunlight. Conversely, the temperature optima for the two enzymes did not differ between alternative radiation environments. The specific activity of POD increased with temperature increase, and was always greater in plants growing under full sunlight than in those from vegetation shade. The activation energy of POD was higher than that of SOD or CAT, being lower in sun-than in shade-exposed plants.


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