Determinants of the Clinical Outcomes of Gout During the First Year of Urate-Lowering Therapy

Clinical benefit early in urate-lowering treatment of gout is difficult to document. We examined data from 1,832 gouty subjects treated with either urate-lowering agents or placebo to identify determinants of gout flare incidence and tophus size during year 1 of treatment. Reductions from pretreatment serum urate levels influenced flare frequency and tophus size, but the effect of urate level on flare incidence was biphasic. Lower urate levels were associated with higher flare incidence early in treatment but lower incidence by one year. The complex relationship between urate-lowering and clinical outcome early in treatment has implications for both clinical and investigative approaches to urate-lowering management.     

Determinants of the clinical outcomes of gout during the first year of urate-lowering therapy

Clinical benefit early in urate-lowering treatment of gout is difficult to document. We examined data from 1,832 gouty subjects treated with either urate-lowering agents or placebo to identify determinants of gout flare incidence and tophus size during year 1 of treatment. Reductions from pretreatment serum urate levels influenced flare frequency and tophus size, but the effect of urate level on flare incidence was biphasic. Lower urate levels were associated with higher flare incidence early in treatment but lower incidence by one year. The complex relationship between urate-lowering and clinical outcome early in treatment has implications for both clinical and investigative approaches to urate-lowering management.     

Urinary Stone Analysis in a Patient with Hyperuricemia to Determine the Mechanism of Stone Formation

In order to determine the mechanism of urinary stone formation in patients with hyperuricemia, we analyzed the crystal components and matrix proteins in a urinary stone from such a patient. Micro-area X-ray spectrometry and infrared (IR) spectroscopy suggested that the outside of the stone was composed of calcium oxalate monohydrate (COM) and the inside of uric acid (UA). Proteomic analysis identified 37 and 14 proteins from the inside and outside of the stone, respectively, as matrix proteins. The proteins that were identified in an ethylenediaminetetraacetic acid (EDTA) fraction were able to bind calcium ions. Thus, calcium-binding proteins may play a significant role in the formation of urinary stones in patients with hyperuricemia.     

Urinary stone analysis in a patient with hyperuricemia to determine the mechanism of stone formation

In order to determine the mechanism of urinary stone formation in patients with hyperuricemia, we analyzed the crystal components and matrix proteins in a urinary stone from such a patient. Micro-area X-ray spectrometry and infrared (IR) spectroscopy suggested that the outside of the stone was composed of calcium oxalate monohydrate (COM) and the inside of uric acid (UA). Proteomic analysis identified 37 and 14 proteins from the inside and outside of the stone, respectively, as matrix proteins. The proteins that were identified in an ethylenediaminetetraacetic acid (EDTA) fraction were able to bind calcium ions. Thus, calcium-binding proteins may play a significant role in the formation of urinary stones in patients with hyperuricemia.     

Syndromic approach to MR diagnosis of gouty arthritis

The objective of the study was to improve radiodiagnosis of gouty arthritis. Thirty-six gout patients aged 39 to 58 years were examined. All the patients underwent X-ray of affected joints and magnetic resonance imaging (MRI). X-ray and MRI revealed tophus formation. The latter also showed articular syndrome and extraarticular manifestations of gout. Analysis of the findings indicated that MRI could enhance the informative value of clinicoradiographic examination of gout patients.     

Jicht in de wervelkolom: een zeldzame presentatie van een veelvoorkomende aandoening

A 92-year old woman was admitted to the geriatric department because of a confusional state due to a urinary tract infection, a severe dermatitis and gout. An incomplete tetraplegia was found on physical examination. Radiological examination showed a large gouty tophus causing spinal cord compression and destruction of the dens. Gouty involvement of the spine is a less common complication of the disease. Optimal treatment of gout is important to prevent the development of such severe complications.     

Absolute protein quantification by LC/MS(E) for global analysis of salicylic acid-induced plant protein secretion responses

The plant cell wall is a dynamic cellular compartment consisting of a complex matrix of components that can change dramatically in response to environmental stresses. During pathogen attack, for instance, a wide spectrum of proteins that participate in various sequential processes involved in plant defense is secreted into the cell wall. In this study, a mass spectrometry, data-independent acquisition approach known as LC/MS (E) was used to assess temporal changes in the cell wall proteome in response to different levels of an endogenous inducer of plant disease defense responses, salicylic acid (SA). LC/MS (E) was used as a label-free method that enabled simultaneous protein identification and absolute femtomole quantification of each protein secreted into the extracellular matrix. A total of 74 secreted proteins were identified, 63 of which showed increased specific secretion in response to SA. A majority of this induced secretion occurred within 2 h of treatment, indicating that many proteins are involved in the early stages of plant defenses. We also identified a number of apparently nonclassically secreted proteins, suggesting that, as in many nonplant systems, Golgi/ER-independent mechanisms exist for plant protein secretion. These results provide new insight into plant apoplastic defense mechanisms and demonstrate that LC/MS (E) is a powerful tool for obtaining both relative and absolute proteome-scale quantification that can be applied to complex, time- and dose-dependent experimental designs.     

Analysis of cotton (Gossypium hirsutum) root proteomes during a compatible interaction with the black root rot fungus Thielaviopsis basicola

A proteomic approach was used to uncover the inducible molecular defense mechanism of cotton root occurring during the compatible interaction with Thielaviopsis basicola. Microscopic observation of cotton root inoculated with a suspension of conidia showed that this necrotrophic hemibiotroph fungus interacts with the plant and completes its life cycle in our experimental system. 2‐DE analysis of root extracts taken after 1, 3, 5, and 7 days postinoculation and cluster analysis of the protein expression levels showed four major profiles (constant, upregulated, one slightly downregulated, and one dramatically downregulated). Spots significantly (p<0.05) upregulated were analyzed by LC‐MS/MS and identified using MASCOT MS/MS ion search software and associated databases. These proteins included defense and stress related proteins, such as pathogenesis‐related proteins and proteins likely to be involved in the oxidative burst, sugar, and nitrogen metabolism as well as amino acid and isoprenoid synthesis. While many of the identified proteins are common components of the defense response of most plants, a proteasome subunit and a protein reported to be induced only in cotton root following Meloidogyne incognita infection were also identified.     

Long-term effects of simulated acid rain stress on a staple forest plant, Pinus massoniana Lamb: a proteomic analysis

At present, acid rain has become one of the top ten global environmental issues. Acid rain causes slower growth, injury, or decline of forests. Some dramatic effects on forests have been observed in south China since the late 1970s and the situation is deteriorating. We carried out a comparative proteomic analysis on Pinus massoniana Lamb, a staple tree species widely distributed in middle and south China to gain a better understanding of tree response to acid rain at molecular level. Two-year-old P. massoniana saplings were treated with simulated AR (SiAR) or control solution, respectively, for 8 months. The changes in total protein profile of P. massoniana leaves were studied using two-dimensional differential gel electrophoresis (2D-DIGE). Among the total protein spots reproducibly detected on each gel, 65 spots representing 28 proteins were identified to be differentially regulated. These proteins were annotated in various biological functions, such as photosynthesis and energy metabolism, secondary metabolism, protein stability, amino acid and nitrogen metabolism and defense. Down-regulation of four key enzymes in the Calvin cycle identified that biomass loss by SiAR was mainly due to the inhibition of carbon fixation. Primary energy metabolisms involved in sucrose biosynthesis, glycolytic pathway and Krebs cycle, etc., were also disturbed after SiAR treatment. Specifically, most of up-regulated proteins were related to secondary metabolism, protein stability and defense, suggesting that in response to SiAR stress, plants started a variety of metabolic pathways to prevent cells from damage. Different from the herbaceous plants suffering SiAR, it revealed that secondary metabolites in P. massoniana play pivotal roles against SiAR. Protemoic techniques were demonstrated a reliable and robust tool to expand our understanding of differentially expressed proteins associated with acid rain stress on P. massoniana. Functional analysis of these proteins further revealed biochemical and physiological basis of the plant in response to acid rain and would provide strategies for breeding new acid rain tolerant tree species. To our knowledge, it is the first proteome report on the forest plant suffering long-term acid rain stress.     

Characterisation of host defence proteins in milk using a proteomic approach

Besides providing nutrition to the newborn, milk also protects the neonate and the mammary gland against infection. As well as the six major proteins, bovine milk contains minor proteins, not all of which have been characterized. In this study, we have subjected bovine skim milk, whey, and milk fat globule membrane (MFGM) fractions to both direct liquid chromatography-tandem mass spectrometry (LC-MS/MS), and two-dimensional electrophoresis (2-DE) followed by matrix assisted laser desorption ionization-time-of-flight (MALDI-TOF) mass spectrometry (MS) of individual protein spots to better characterize the repertoire of minor milk proteins, particularly those involved with host defense. Milk from peak lactation as well as during the period of colostrum formation and during mastitis were analyzed to gain a more complete sampling of the milk proteome. In total, 2903 peptides were detected by LC-MS and 2770 protein spots by 2-DE. From these, 95 distinct gene products were identified, comprising 53 identified through direct LC-MS/MS and 57 through 2-DE-MS. The latter were derived from a total of 363 spots analyzed with 181 being successfully identified. At least 15 proteins were identified that are involved in host defense. These results demonstrate that the proteome of milk is more complex than has previously been reported and a significant fraction of minor milk proteins are involved in protection against infection.     

New targets of Arabidopsis thioredoxins revealed by proteomic analysis

Proteomics was used to search for putative thioredoxin (TRX) targets in leaves of the model plant, Arabidopsis thaliana. About forty different proteins have been found to be reduced by TRX, after TRX itself has been specifically reduced by its NADPH-dependent reductase. Twenty-one of the identified proteins were already known or recently proposed to be TRX-dependent and nineteen of the proteins were new potential targets. The identified proteins are involved in a wide variety of processes, including the Calvin cycle, metabolism, photosynthesis, folding, defense against oxidative stress and amino acid synthesis. Two proteins from the glycine cleavage complex were also identified as putative TRX targets, and a new role can be postulated in leaves for TRX in defense against herbivores and/or pathogens.     

Identification of human plasma C1 inhibitor as a target protein for staphylococcal superantigen-like protein 5 (SSL5)

The family of staphylococcal superantigen-like proteins (SSLs) have a structure similar to bacterial superantigens but exhibit no superantigenic activity. These exoproteins have recently been shown to disturb the host immune defense system. One family member, SSL5, was reported to bind to human leukocyte P-selectin glycoprotein ligand-1 (PSGL-1) and matrix metalloproteinase-9 (MMP-9) and to interfere with leukocyte trafficking. In the present study, we explored human plasma proteins bound by glutathione S-transferase (GST)-tagged recombinant SSL5 (GST-SSL5) and identified plasma protease C1 inhibitor (C1Inh) as a major SSL5-binding protein based on the results of peptide mass fingerprinting analysis with MALDI-TOFMS. GST-SSL5 was found to attenuate the inhibitory activity of recombinant histidine-tagged C1Inh (C1Inh-His) toward complement C1s. We also observed that the treatment of C1Inh-His with neuraminidase markedly decreased its binding to GST-SSL5. Moreover, C1Inh-His produced by Lec2 mutant cells (deficient in sialic acid biosynthesis) showed much lower binding affinity for SSL5 than that produced by the wild-type CHO-K1 cells, as assessed by pull-down assay. These results suggest that SSL5 binds to C1Inh in a sialic acid-dependent fashion and modulates the host immune defense through perturbation of the complement system in association with S. aureus infection.     

Changes in the leaf proteome profile of Mentha arvensis in response to Alternaria alternata infection

Plant pathogenic fungi cause important yield losses in crops. A proteomic approach was used to study the changes in the leaf proteome profile of the plant Mentha arvensis infected with a necrotrophic fungus, Alternaria alternata. High-resolution two-dimensional gel electrophoresis (2-DE) followed by colloidal Coomassie staining and mass spectrometric analysis was used to identify highly abundant proteins differentially expressed in response to fungal infection. From a total of 210 reproducibly detected and analyzed spots, the intensity of sixty-seven spots was altered, and forty-five of them were successfully identified by matrix assisted laser desorption/ionization time of flight-mass spectrometry (MALDI TOF/TOF MS/MS). Fifty-six percent of the identified proteins belonged to energy and metabolism whereas 29% were stress and defense related. Taken together, the results allow to assess changes at the proteomic level in the host due to the defense response. Results show an initial defense response, not strong enough to overcome the pathogenesis, which may be similar to other susceptible plant-pathogen interactions; however, cross-talks between various defense pathways, regulatory networks and physiological conditions are other important aspects to be considered.     

Proteomic analysis of the venom of the predatory ant Pachycondyla striata (Hymenoptera: Formicidae)

The ants use their venom for predation, defense, and communication. The venom of these insects is rich in peptides and proteins, and compared with other animal venoms, ant venoms remain poorly explored. The objective of this study was to evaluate the protein content of the venom in the Ponerinae ant Pachycondyla striata. Venom samples were collected by manual gland reservoir dissection, and samples were submitted to two‐dimensional gel electrophoresis and separation by ion‐exchange and reverse‐phase high‐performance liquid chromatography followed by mass spectrometry using tanden matrix‐assisted laser desorption/ionization with time‐of‐flight (MALDI‐TOF/TOF) mass spectrometry and electrospray ionization‐quadrupole with time‐of‐flight (ESI‐Q/TOF) mass spectrometry for obtaining amino acid sequence. Spectra obtained were searched against the NCBInr and SwissProt database. Additional analysis was performed using PEAKS Studio 7.0 (Sequencing de novo). The venom of P. striata has a complex mixture of proteins from which 43 were identified. Within the identified proteins are classical venom proteins (phospholipase A, hyaluronidase, and aminopeptidase N), allergenic proteins (different venom allergens), and bioactive peptides (U10‐ctenitoxin Pn1a). Venom allergens are among the most expressed proteins, suggesting that P. striata venom has high allergenic potential. This study discusses the possible functions of the proteins identified in the venom of P. striata.     

Comprehensive proteome analysis of lettuce latex using multidimensional protein-identification technology

Commercially, lettuce (Lactuca sativa) is one of the most important leafy vegetables. Lettuce produces a milky latex of variable chemical compositions within its laticifers. As a step toward understanding the main physiological roles of this latex in higher plants, we embarked on its proteomic analysis. We investigated 587 latex proteins that were identified from the lettuce latex using multidimensional protein-identification technology. A bioinformatics analysis showed that the most frequently encountered proteins in the latex were organellar proteins from plastids and mitochondria, followed by nucleic and cytoplasmic proteins. Functional classification of the identified proteins showed that proteins related to metabolism, cell rescue, defense, and virulence were the most abundant in lettuce latex. Furthermore, numerous resistance proteins of lettuce and viral proteins were present in the latex suggesting for the first time a possible function of the lettuce latex in defense or pathogenesis. To the knowledge of the authors, this is the first large-scale proteome analysis of lettuce latex.