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1.
Molecules ; 29(16)2024 Aug 16.
Article in English | MEDLINE | ID: mdl-39202966

ABSTRACT

The Japanese plum tree (Prunus salicina Lindl.) is mainly cultivated in temperate areas of China and some European countries. Certain amounts of wood (from pruning works) are generated every year from this crop of worldwide commercial significance. The main objective of this work was to value this agricultural woody residue, for which the chemical composition of pruning wood extracts from six Japanese plum cultivars was investigated, and the antiproliferative activity of extracts and pure phenolics present in those extracts was measured. For the chemical characterization, total phenolic content and DPPH radical-scavenging assays and HPLC‒DAD/ESI‒MS analyses were performed, with the procyanidin (-)-ent-epicatechin-(2α→O→7,4α→8)-epicatechin (5) and the propelargonidin (+)-epiafzelechin-(2ß→O→7,4ß→8)-epicatechin (7) being the major components of the wood extracts. Some quantitative differences were found among plum cultivars, and the content of proanthocyanidins ranged from 1.50 (cv. 'Fortune') to 4.44 (cv. 'Showtime') mg/g of dry wood. Regarding the antitumoral activity, eight wood extracts and four phenolic compounds were evaluated in MCF-7 cells after 48 h of induction, showing the wood extract from cv. 'Songold' and (‒)-annphenone (3), the best antiproliferative activity (IC50: 424 µg/mL and 405 µg/mL, respectively).


Subject(s)
Plant Extracts , Wood , Humans , Wood/chemistry , Plant Extracts/pharmacology , Plant Extracts/chemistry , Cell Proliferation/drug effects , Antineoplastic Agents, Phytogenic/pharmacology , Antineoplastic Agents, Phytogenic/chemistry , Prunus domestica/chemistry , Phenols/chemistry , Phenols/pharmacology , Phenols/analysis , Chromatography, High Pressure Liquid , MCF-7 Cells , Prunus/chemistry , Cell Line, Tumor , Antineoplastic Agents/pharmacology , Antineoplastic Agents/chemistry , Antioxidants/pharmacology , Antioxidants/chemistry
2.
Environ Res ; 229: 116001, 2023 07 15.
Article in English | MEDLINE | ID: mdl-37116679

ABSTRACT

Propyl-propane-thiosulfonate (PTSO) is an organosulfur compound found inAllium spp. Due to its antioxidant and antimicrobial activities, PTSO has been proposed for applications in the agri-food sector, such as feed additive. However, its use with commercial purposes depends on its toxicity evaluation. The present work aimed to perform a pilot-study of toxicokinetic profile of PTSO combining in silico and in vitro techniques, important steps in the risk assessment process. In silico ecotoxicity studies were also performed considering the importance of the environmental impact of the compound before its commercial use. First, an analytical method has been developed and validated to determine the original compound and its metabolites by ultra-performance liquid chromatography-tandem mass spectrometry. The phase I and II metabolism of PTSO was predicted using Meta-Pred Web Server. For the phase I metabolism, rat (male and female) and human liver microsomes were incubated with PTSO and NADPH regeneration system. Furthermore, in the phase II, microsomes were incubated with PTSO and glutathione or uridine 5'- diphosphoglucuronic acid. The analysis revealed the presence of propylpropane thiosulfinate (PTS) originated by redox reaction in phase I, and two conjugates from the phase II: S-propylmercaptoglutathione (GSSP) and S-propylmercaptocysteine (CSSP). Additionally, considering the environmental fate of PTSO and its metabolites, the ADME parameters and the potential ecotoxicity were also predicted using in silico softwares. The results of the ecotoxicity in silico study evidenced that the metabolism induced the formation of detoxified metabolites from the parent compound, except for dimercaprol and 3-mercaptopropane1,2-diol. Further in vivo assays are needed to confirm this prediction.


Subject(s)
Allium , Male , Rats , Humans , Female , Animals , Allium/chemistry , Pilot Projects , Antioxidants , Microsomes, Liver , Chromatography, High Pressure Liquid
3.
Int J Mol Sci ; 24(11)2023 Jun 01.
Article in English | MEDLINE | ID: mdl-37298563

ABSTRACT

We explored the genome of the Wolbachia strain, wEsol, symbiotic with the plant-gall-inducing fly Eurosta solidaginis with the goal of determining if wEsol contributes to gall induction by its insect host. Gall induction by insects has been hypothesized to involve the secretion of the phytohormones cytokinin and auxin and/or proteinaceous effectors to stimulate cell division and growth in the host plant. We sequenced the metagenome of E. solidaginis and wEsol and assembled and annotated the genome of wEsol. The wEsol genome has an assembled length of 1.66 Mbp and contains 1878 protein-coding genes. The wEsol genome is replete with proteins encoded by mobile genetic elements and shows evidence of seven different prophages. We also detected evidence of multiple small insertions of wEsol genes into the genome of the host insect. Our characterization of the genome of wEsol indicates that it is compromised in the synthesis of dimethylallyl pyrophosphate (DMAPP) and S-adenosyl L-methionine (SAM), which are precursors required for the synthesis of cytokinins and methylthiolated cytokinins. wEsol is also incapable of synthesizing tryptophan, and its genome contains no enzymes in any of the known pathways for the synthesis of indole-3-acetic acid (IAA) from tryptophan. wEsol must steal DMAPP and L-methionine from its host and therefore is unlikely to provide cytokinin and auxin to its insect host for use in gall induction. Furthermore, in spite of its large repertoire of predicted Type IV secreted effector proteins, these effectors are more likely to contribute to the acquisition of nutrients and the manipulation of the host's cellular environment to contribute to growth and reproduction of wEsol than to aid E. solidaginis in manipulating its host plant. Combined with earlier work that shows that wEsol is absent from the salivary glands of E. solidaginis, our results suggest that wEsol does not contribute to gall induction by its host.


Subject(s)
Tephritidae , Wolbachia , Animals , Wolbachia/genetics , Tryptophan , Tephritidae/metabolism , Insecta/metabolism , Indoleacetic Acids/metabolism , Cytokinins , Genomics
4.
Sensors (Basel) ; 22(19)2022 Oct 09.
Article in English | MEDLINE | ID: mdl-36236738

ABSTRACT

Hyperspectral imaging opens up new opportunities for masked face recognition via discrimination of the spectral information obtained by hyperspectral sensors. In this work, we present a novel algorithm to extract facial spectral-features from different regions of interests by performing computer vision techniques over the hyperspectral images, particularly Histogram of Oriented Gradients. We have applied this algorithm over the UWA-HSFD dataset to extract the facial spectral-features and then a set of parallel Support Vector Machines with custom kernels, based on the cosine similarity and Euclidean distance, have been trained on fly to classify unknown subjects/faces according to the distance of the visible facial spectral-features, i.e., the regions that are not concealed by a face mask or scarf. The results draw up an optimal trade-off between recognition accuracy and compression ratio in accordance with the facial regions that are not occluded.


Subject(s)
Facial Recognition , Algorithms , Support Vector Machine
5.
Neurol Sci ; 42(1): 215-223, 2021 Jan.
Article in English | MEDLINE | ID: mdl-32617742

ABSTRACT

INTRODUCTION: The aim of this study is to describe the normal cross-sectional area (CSA) and appearance of cervical nerve roots in ultrasound, correlating it to age and other patient somatic parameters. METHODS: One hundred healthy volunteers were included. We aimed to achieve uniform representation throughout all age groups. Ultrasound of the cervical nerve roots was performed bilaterally. CSA and margins description were obtained. RESULTS: C5 nerve, 8.32 ± 2.30; C6 nerve, 11.88 ± 3.36; C7 nerve, 12.79 ± 3.85; C8 nerve, 11.20 ± 3.45. Significant correlation between CSA and age was demonstrated, but not for body mass index. Blurred margins were present in up to 23.71% cervical nerves, more frequently in older individuals and in C7 nerve. DISCUSSION: If ultrasound morphology of cervical nerve roots is used as a diagnostic parameter, the normal range of CSA values and percentage of blurred margins according to age should be considered.


Subject(s)
Cervical Vertebrae , Spinal Nerve Roots , Aged , Body Mass Index , Cervical Vertebrae/diagnostic imaging , Healthy Volunteers , Humans , Reference Values , Spinal Nerve Roots/diagnostic imaging , Ultrasonography
6.
Andrologia ; 53(8): e14151, 2021 Sep.
Article in English | MEDLINE | ID: mdl-34170025

ABSTRACT

We present the case of a 48 year-old man with a history of pulmonary tuberculosis who was brought to our centre with a progressively increasing left testicular mass. Radiological findings were not conclusive on whether the mass was neoplastic or infectious in nature. Therefore, an orchiectomy was performed. Pathology examination revealed coccidioidomycosis of the left testicle, highlighting an exceedingly rare presentation of disseminated genitourinary coccidioidomycosis.


Subject(s)
Coccidioidomycosis , Neoplasms, Germ Cell and Embryonal , Testicular Neoplasms , Coccidioidomycosis/diagnosis , Humans , Male , Middle Aged , Orchiectomy , Testicular Neoplasms/diagnostic imaging , Testicular Neoplasms/surgery
7.
Int J Biometeorol ; 65(1): 107-117, 2021 Jan.
Article in English | MEDLINE | ID: mdl-32198562

ABSTRACT

The term neurodegenerative diseases include a long list of diseases affecting the nervous system that are characterized by the degeneration of different neurological structures. Among them, Alzheimer disease (AD), Parkinson disease (PD), and amyotrophic lateral sclerosis (ALS) are the most representative ones. The vast majority of cases are sporadic and results from the interaction of genes and environmental factors in genetically predisposed individuals. Among environmental conditions, electromagnetic field exposure has begun to be assessed as a potential risk factor for neurodegeneration. In this review, we discuss the existing literature regarding electromagnetic fields and neurodegenerative diseases. Epidemiological studies in AD, PD, and ALS have shown discordant results; thus, a clear correlation between electromagnetic exposure and neurodegeneration has not been demonstrated. In addition, we discuss the role of electromagnetic radiation as a potential non-invasive therapeutic strategy for some neurodegenerative diseases, particularly for PD and AD.


Subject(s)
Alzheimer Disease , Amyotrophic Lateral Sclerosis , Neurodegenerative Diseases , Parkinson Disease , Alzheimer Disease/epidemiology , Amyotrophic Lateral Sclerosis/epidemiology , Electromagnetic Fields , Humans , Neurodegenerative Diseases/epidemiology
8.
Chemphyschem ; 21(12): 1258-1271, 2020 06 16.
Article in English | MEDLINE | ID: mdl-32352214

ABSTRACT

The thermoresponsive nature of aqueous solutions of poly(N-isopropylacrylamide) (PNIPAAM) star polymers containing 2, 3, 4, and 6 arms has been investigated by turbidity, dynamic light scattering, rheology, and rheo-SALS. Simulations of the thermosensitive nature of the single star polymers have also been conducted. Some of the samples form aggregates even at temperatures significantly below the lower critical solution temperature (LCST) of PNIPAAM. Increasing concentration and number of arms promotes associations at low temperatures. When the temperature is raised, there is a competition between size increase due to enhanced aggregation and a size reduction caused by contraction. Monte Carlo simulations show that the single stars contract with increasing temperature, and that this contraction is more pronounced when the number of arms is increased. Some samples exhibit a minimum in the turbidity data after the initial increase at the cloud point. The combined rheology and rheo-SALS data suggest that this is due to a fragmentation of the aggregates followed by re-aggregation at even higher temperatures. Although the 6-arm star polymer aggregates more than the other stars at low temperatures, the more compact structure renders it less prone to aggregation at temperatures above the cloud point.

9.
Neurol Sci ; 41(12): 3779-3781, 2020 Dec.
Article in English | MEDLINE | ID: mdl-32955639

ABSTRACT

Myoclonus-dystonia associated with epsilon-sarcoglycan gene (SGCE) is a rare disorder characterized by myoclonus involving the upper body (neck, trunk, upper limbs) and proximal muscles associated with dystonia in more than half of the patients. When the clinical picture is clearly identified, more than half of the cases are associated with mutations in the SGCE gene. We herein describe a family with myoclonus-dystonia associated with a novel mutation in exon 7 of SGCE, c.904A>T (p.Lys302Ter) [Chr7:(GRCh38):g.94600779 T>A], which was absent in a non-affected member. A video recording of two of the affected members is provided. While the index case presents a severe cervical dystonia even affecting back posture, his sibling shows a much milder phenotype with mild myoclonic jerks. None of them had alcohol responsiveness or psychiatric comorbidity.


Subject(s)
Dystonic Disorders , Myoclonus , Biological Variation, Population , Dystonic Disorders/genetics , Humans , Mutation/genetics , Myoclonus/complications , Myoclonus/genetics , Sarcoglycans/genetics
10.
Proc Natl Acad Sci U S A ; 114(12): 3085-3090, 2017 03 21.
Article in English | MEDLINE | ID: mdl-28265087

ABSTRACT

Active-learning pedagogies have been repeatedly demonstrated to produce superior learning gains with large effect sizes compared with lecture-based pedagogies. Shifting large numbers of college science, technology, engineering, and mathematics (STEM) faculty to include any active learning in their teaching may retain and more effectively educate far more students than having a few faculty completely transform their teaching, but the extent to which STEM faculty are changing their teaching methods is unclear. Here, we describe the development and application of the machine-learning-derived algorithm Decibel Analysis for Research in Teaching (DART), which can analyze thousands of hours of STEM course audio recordings quickly, with minimal costs, and without need for human observers. DART analyzes the volume and variance of classroom recordings to predict the quantity of time spent on single voice (e.g., lecture), multiple voice (e.g., pair discussion), and no voice (e.g., clicker question thinking) activities. Applying DART to 1,486 recordings of class sessions from 67 courses, a total of 1,720 h of audio, revealed varied patterns of lecture (single voice) and nonlecture activity (multiple and no voice) use. We also found that there was significantly more use of multiple and no voice strategies in courses for STEM majors compared with courses for non-STEM majors, indicating that DART can be used to compare teaching strategies in different types of courses. Therefore, DART has the potential to systematically inventory the presence of active learning with ∼90% accuracy across thousands of courses in diverse settings with minimal effort.


Subject(s)
Problem-Based Learning/standards , Science/education , Teaching/standards , Humans , Sound , Students , Technology , Universities/standards
11.
Environ Microbiol ; 21(6): 1980-1988, 2019 06.
Article in English | MEDLINE | ID: mdl-30370610

ABSTRACT

Marine Group I (MGI) Thaumarchaeota are some of the most abundant microorganisms in the deep ocean and responsible for much of the ammonia oxidation occurring in this environment. In this work, we present 35 sequences assembled from metagenomic samples of the first uncultivated Caudovirales viruses associated with Thaumarchaeota, which we designated marthavirus. Most of the sequences were obtained from cellular metagenomes confirming that they represent an important tool to study environmental viral communities due to cells retrieved while undergoing viral lysis. Metagenomic recruitment showed that this viral population is formed by very divergent entities with high intrapopulation homogeneity. However, metatranscriptomic analyses revealed the same differential expression profile with the capsid as major transcript, indicative of viruses during the lytic cycle. The cobalamine biosynthesis gene cobS, an auxiliary metabolic gene, was also highly expressed during the infection. These analyses expand our understanding of the global diversity of archaeal viruses.


Subject(s)
Archaea/virology , Archaeal Viruses/isolation & purification , Caudovirales/isolation & purification , Archaea/genetics , Archaeal Viruses/classification , Archaeal Viruses/genetics , Caudovirales/classification , Caudovirales/genetics , Genome, Viral , Metagenome , Phylogeny
12.
Int J Mol Sci ; 19(7)2018 06 24.
Article in English | MEDLINE | ID: mdl-29937518

ABSTRACT

The LrtA protein of Synechocystis sp. PCC 6803 intervenes in cyanobacterial post-stress survival and in stabilizing 70S ribosomal particles. It belongs to the hibernating promoting factor (HPF) family of proteins, involved in protein synthesis. In this work, we studied the conformational preferences and stability of isolated LrtA in solution. At physiological conditions, as shown by hydrodynamic techniques, LrtA was involved in a self-association equilibrium. As indicated by Nuclear Magnetic Resonance (NMR), circular dichroism (CD) and fluorescence, the protein acquired a folded, native-like conformation between pH 6.0 and 9.0. However, that conformation was not very stable, as suggested by thermal and chemical denaturations followed by CD and fluorescence. Theoretical studies of its highly-charged sequence suggest that LrtA had a Janus sequence, with a context-dependent fold. Our modelling and molecular dynamics (MD) simulations indicate that the protein adopted the same fold observed in other members of the HPF family (ß-α-ß-ß-ß-α) at its N-terminal region (residues 1­100), whereas the C terminus (residues 100­197) appeared disordered and collapsed, supporting the overall percentage of overall secondary structure obtained by CD deconvolution. Then, LrtA has a chameleonic sequence and it is the first member of the HPF family involved in a self-association equilibrium, when isolated in solution.


Subject(s)
Bacterial Proteins/chemistry , Ribosomal Proteins/chemistry , Ribosomes/chemistry , Synechocystis/chemistry , Amino Acid Motifs , Bacterial Proteins/genetics , Bacterial Proteins/metabolism , Cloning, Molecular , Escherichia coli/genetics , Escherichia coli/metabolism , Gene Expression , Genetic Vectors/chemistry , Genetic Vectors/metabolism , Molecular Dynamics Simulation , Protein Conformation, alpha-Helical , Protein Conformation, beta-Strand , Protein Folding , Protein Interaction Domains and Motifs , Protein Multimerization , Protein Stability , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Ribosomal Proteins/genetics , Ribosomal Proteins/metabolism , Ribosomes/metabolism , Solutions , Synechocystis/metabolism , Thermodynamics
13.
Environ Microbiol ; 19(7): 2681-2700, 2017 07.
Article in English | MEDLINE | ID: mdl-28419726

ABSTRACT

Thaumarchaeota are globally distributed and abundant microorganisms occurring in diverse habitats and thus represent a major source of archaeal lipids. The scope of lipids as taxonomic markers in microbial ecological studies is limited by the scarcity of comparative data on the membrane lipid composition of cultivated representatives, including the phylum Thaumarchaeota. Here, we comprehensively describe the core and intact polar lipid (IPL) inventory of ten ammonia-oxidising thaumarchaeal cultures representing all four characterized phylogenetic clades. IPLs of these thaumarchaeal strains are generally similar and consist of membrane-spanning, glycerol dibiphytanyl glycerol tetraethers with monoglycosyl, diglycosyl, phosphohexose and hexose-phosphohexose headgroups. However, the relative abundances of these IPLs and their core lipid compositions differ systematically between the phylogenetic subgroups, indicating high potential for chemotaxonomic distinction of thaumarchaeal clades. Comparative lipidomic analyses of 19 euryarchaeal and crenarchaeal strains suggested that the lipid methoxy archaeol is synthesized exclusively by Thaumarchaeota and may thus represent a diagnostic lipid biomarker for this phylum. The unprecedented diversity of the thaumarchaeal lipidome with 118 different lipids suggests that membrane lipid composition and adaptation mechanisms in Thaumarchaeota are more complex than previously thought and include unique lipids with as yet unresolved properties.


Subject(s)
Archaea/metabolism , Glyceryl Ethers/analysis , Membrane Lipids/analysis , Archaea/classification , Archaea/genetics , Biomarkers/analysis , Ecosystem , Geologic Sediments/microbiology , Phylogeny , Soil Microbiology , Water Microbiology
14.
Annu Rev Microbiol ; 66: 83-101, 2012.
Article in English | MEDLINE | ID: mdl-22994489

ABSTRACT

The discovery of ammonia-oxidizing archaea (AOA), now generally recognized to exert primary control over ammonia oxidation in terrestrial, marine, and geothermal habitats, necessitates a reassessment of the nitrogen cycle. In particular, the unusually high affinity of marine and terrestrial AOA for ammonia indicates that this group may determine the oxidation state of nitrogen available to associated micro- and macrobiota, altering our current understanding of trophic interactions. Initial comparative genomics and physiological studies have revealed a novel, and as yet unresolved, primarily copper-based pathway for ammonia oxidation and respiration distinct from that of known ammonia-oxidizing bacteria and possibly relevant to the production of atmospherically active nitrogen oxides. Comparative studies also provide compelling evidence that the lineage of Archaea with which the AOA affiliate is sufficiently divergent to justify the creation of a novel phylum, the Thaumarchaeota.


Subject(s)
Ammonia/metabolism , Archaea/genetics , Archaea/physiology , Genetic Variation , Metabolic Networks and Pathways , Nitrogen/metabolism , Oxidation-Reduction
16.
Angew Chem Int Ed Engl ; 56(3): 782-785, 2017 01 16.
Article in English | MEDLINE | ID: mdl-27933691

ABSTRACT

A new electrochemical framework for tracking individual soft particles in solution and monitoring their fusion with polarized liquid-liquid interfaces is reported. The physicochemical principle lies in the interfacial transfer of an ionic probe confined in the particles dispersed in solution and that is released upon their collision and fusion with the fluid interface. As a proof-of-concept, spike-like transients of a stochastic nature are reported in the current-time response of 1,2-dichloroethane(DCE)|water(W) submilli-interfaces after injection of DCE-in-W emulsions. The sign and potential dependence of the spikes reflect the charge and lipophilicity of the ionic load of the droplets. A comparison with dynamic light scattering measurements indicates that each spike is associated with the collision of a single sub-picoliter droplet. This opens a new framework for the study of single fusion events at the micro- and nanoscale and of ion transport across biomimetic soft interfaces.

17.
Biochemistry ; 55(45): 6209-6220, 2016 Nov 15.
Article in English | MEDLINE | ID: mdl-27791355

ABSTRACT

The assembly of the protein complex of cytochrome c oxidase (COX), which participates in the mitochondrial respiratory chain, requires a large number of accessory proteins (the so-called assembly factors). Human COX assembly factor 3 (hCOA3), also known as MITRAC12 or coiled-coil domain-containing protein 56 (CCDC56), interacts with the first subunit protein of COX to form its catalytic core and promotes its assemblage with the other units. Therefore, hCOA3 is involved in COX biogenesis in humans and can be exploited as a drug target in patients with mitochondrial dysfunctions. However, to be considered a molecular target, its structure and conformational stability must first be elucidated. We have embarked on the description of such features by using spectroscopic and hydrodynamic techniques, in aqueous solution and in the presence of detergents, together with computational methods. Our results show that hCOA3 is an oligomeric protein, forming aggregates of different molecular masses in aqueous solution. Moreover, on the basis of fluorescence and circular dichroism results, the protein has (i) its unique tryptophan partially shielded from solvent and (ii) a relatively high percentage of secondary structure. However, this structure is highly flexible and does not involve hydrogen bonding. Experiments in the presence of detergents suggest a slightly higher content of nonrigid helical structure. Theoretical results, based on studies of the primary structure of the protein, further support the idea that hCOA3 is a disordered protein. We suggest that the flexibility of hCOA3 is crucial for its interaction with other proteins to favor mitochondrial protein translocation and assembly of proteins involved in the respiratory chain.


Subject(s)
Membrane Proteins/chemistry , Mitochondrial Proteins/chemistry , Protein Multimerization , Protein Structure, Secondary , Solutions/chemistry , Amino Acid Sequence , Circular Dichroism , Computer Simulation , Electron Transport Complex IV/chemistry , Electron Transport Complex IV/metabolism , Humans , Hydrogen-Ion Concentration , Intrinsically Disordered Proteins/chemistry , Intrinsically Disordered Proteins/genetics , Intrinsically Disordered Proteins/metabolism , Kinetics , Membrane Proteins/genetics , Membrane Proteins/metabolism , Mitochondrial Proteins/genetics , Mitochondrial Proteins/metabolism , Models, Molecular , Protein Aggregates , Protein Binding , Protein Domains , Protein Subunits/chemistry , Protein Subunits/metabolism , Sodium Dodecyl Sulfate/chemistry
18.
J Biol Chem ; 290(38): 23077-93, 2015 Sep 18.
Article in English | MEDLINE | ID: mdl-26251518

ABSTRACT

Lysine acetylation is an important post-translational modification in the metabolic regulation of both prokaryotes and eukaryotes. In Escherichia coli, PatZ (formerly YfiQ) is the only known acetyltransferase protein and is responsible for acetyl-CoA synthetase acetylation. In this study, we demonstrated PatZ-positive cooperativity in response to acetyl-CoA and the regulation of acetyl-CoA synthetase activity by the acetylation level. Furthermore, functional analysis of an E809A mutant showed that the conserved glutamate residue is not relevant for the PatZ catalytic mechanism. Biophysical studies demonstrated that PatZ is a stable tetramer in solution and is transformed to its octameric form by autoacetylation. Moreover, this modification is reversed by the sirtuin CobB. Finally, an in silico PatZ tetramerization model based on hydrophobic and electrostatic interactions is proposed and validated by three-dimensional hydrodynamic analysis. These data reveal, for the first time, the structural regulation of an acetyltransferase by autoacetylation in a prokaryotic organism.


Subject(s)
Acetyltransferases/chemistry , Escherichia coli Proteins/chemistry , Escherichia coli/enzymology , Models, Molecular , Protein Multimerization , Acetylation , Acetyltransferases/genetics , Acetyltransferases/metabolism , Amino Acid Substitution , Animals , Coenzyme A Ligases/chemistry , Coenzyme A Ligases/genetics , Coenzyme A Ligases/metabolism , Escherichia coli/genetics , Escherichia coli Proteins/genetics , Escherichia coli Proteins/metabolism , Humans , Mice , Mutation, Missense , Protein Structure, Quaternary , Sirtuins/chemistry , Sirtuins/genetics , Sirtuins/metabolism
19.
Eur Biophys J ; 45(1): 45-54, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26596272

ABSTRACT

Mucins are the primary macromolecular component of mucus--nature's natural lubricant--although they are poorly characterised heterogeneous substances. Recent advances in hydrodynamic methodology now offer the opportunity for gaining a better understanding of their solution properties. In this study a combination of such methods was used to provide increased understanding of a preparation of porcine intestinal mucin (PIM), MUC2 mucin, in terms of both heterogeneity and quantification of conformational flexibility. The new sedimentation equilibrium algorithm SEDFIT-MSTAR is applied to yield a weight average (over the whole distribution) molar mass of 7.1 × 10(6) g mol(-1), in complete agreement with size exclusion chromatography coupled with multi-angle light scattering (SEC-MALS), which yielded a value of 7.2 × 10(6) g mol(-1). Sedimentation velocity profiles show mucin to be very polydisperse, with a broad molar mass distribution obtained using the Extended Fujita algorithm, consistent with the elution profiles from SEC-MALS. On-line differential pressure viscometry coupled to the SEC-MALS was used to obtain the intrinsic viscosity [η] as a function of molar mass. These data combined with sedimentation coefficient data into the global conformation algorithm HYDFIT show that PIM has a flexible linear structure, with persistence length L p ~10 nm and mass per unit length, M L ~2380 g mol(-1) nm(-1), consistent with a Wales-van Holde ratio of ~1.2 obtained from the concentration dependence of the sedimentation coefficient.


Subject(s)
Algorithms , Hydrodynamics , Mucin-2/chemistry , Animals , Fractionation, Field Flow/methods , Intestinal Mucosa/metabolism , Solutions , Swine
20.
Environ Microbiol ; 17(5): 1600-14, 2015 May.
Article in English | MEDLINE | ID: mdl-25142282

ABSTRACT

Archaea can respond to changes in the environment by altering the composition of their membrane lipids, for example, by modification of the abundance and composition of glycerol dialkyl glycerol tetraethers (GDGTs). Here, we investigated the abundance and proportions of polar GDGTs (P-GDGTs) and core GDGTs (C-GDGTs) sampled in different seasons from Tengchong hot springs (Yunnan, China), which encompassed a pH range of 2.5-10.1 and a temperature range of 43.7-93.6°C. The phylogenetic composition of the archaeal community (reanalysed from published work) divided the Archaea in spring sediment samples into three major groups that corresponded with spring pH: acidic, circumneutral and alkaline. Cluster analysis showed correlation between spring pH and the composition of P- and C-GDGTs and archaeal 16S rRNA genes, indicating an intimate link between resident Archaea and the distribution of P- and C-GDGTs in Tengchong hot springs. The distribution of GDGTs in Tengchong springs was also significantly affected by temperature; however, the relationship was weaker than with pH. Analysis of published datasets including samples from Tibet, Yellowstone and the US Great Basin hot springs revealed a similar relationship between pH and GDGT content. Specifically, low pH springs had higher concentrations of GDGTs with high numbers of cyclopentyl rings than neutral and alkaline springs, which is consistent with the predominance of high cyclopentyl ring-characterized Sulfolobales and Thermoplasmatales present in some of the low pH springs. Our study suggests that the resident Archaea in these hot springs are acclimated if not adapted to low pH by their genetic capacity to effect the packing density of their membranes by increasing cyclopentyl rings in GDGTs at the rank of community.


Subject(s)
Archaea/metabolism , Geologic Sediments/microbiology , Glyceryl Ethers/metabolism , Hot Springs/microbiology , Membrane Lipids/metabolism , Archaea/genetics , Desulfurococcales/genetics , Desulfurococcales/isolation & purification , Environment , Glyceryl Ethers/analysis , Hydrogen-Ion Concentration , Membrane Lipids/analysis , Oxygen/metabolism , Phylogeny , RNA, Ribosomal, 16S/genetics , Seasons , Soil Microbiology , Sulfolobales/genetics , Sulfolobales/isolation & purification , Temperature , Thermoplasmales/genetics , Thermoplasmales/isolation & purification , Tibet
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