scholarly journals TcI Isolates of Trypanosoma cruzi Exploit the Antioxidant Network for Enhanced Intracellular Survival in Macrophages and Virulence in Mice

2016 ◽  
Vol 84 (6) ◽  
pp. 1842-1856 ◽  
Author(s):  
María Paola Zago ◽  
Yashoda M. Hosakote ◽  
Sue-jie Koo ◽  
Monisha Dhiman ◽  
María Dolores Piñeyro ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Biological Behavior ◽  
Data Sets ◽  
Proteomic Profiling ◽  
Two Dimensional Gel Electrophoresis ◽  
Content Type ◽  
Transmission Cycle ◽  
Protein Levels ◽  
Flight Mass Spectrometry ◽  
And Control

Trypanosoma cruzispecies is categorized into six discrete typing units (TcI to TcVI) of which TcI is most abundantly noted in the sylvatic transmission cycle and considered the major cause of human disease. In our study, the TcI strains Colombiana (COL), SylvioX10/4 (SYL), and a cultured clone (TCC) exhibited different biological behavior in a murine model, ranging from high parasitemia and symptomatic cardiomyopathy (SYL), mild parasitemia and high tissue tropism (COL), to no pathogenicity (TCC). Proteomic profiling of the insect (epimastigote) and infective (trypomastigote) forms by two-dimensional gel electrophoresis/matrix-assisted laser desorption ionization–time of flight mass spectrometry, followed by functional annotation of the differential proteome data sets (≥2-fold change,P< 0.05), showed that several proteins involved in (i) cytoskeletal assembly and remodeling, essential for flagellar wave frequency and amplitude and forward motility of the parasite, and (ii) the parasite-specific antioxidant network were enhanced in COL and SYL (versus TCC) trypomastigotes. Western blotting confirmed the enhanced protein levels of cytosolic and mitochondrial tryparedoxin peroxidases and their substrate (tryparedoxin) and iron superoxide dismutase in COL and SYL (versus TCC) trypomastigotes. Further, COL and SYL (but not TCC) were resistant to exogenous treatment with stable oxidants (H2O2and peroxynitrite [ONOO−]) and dampened the intracellular superoxide and nitric oxide response in macrophages, and thus these isolates escaped from macrophages. Our findings suggest that protein expression conducive to increase in motility and control of macrophage-derived free radicals provides survival and persistence benefits to TcI isolates ofT. cruzi.

scholarly journals Proteomic Analysis of Trichoderma atroviride Reveals Independent Roles for Transcription Factors BLR-1 and BLR-2 in Light and Darkness

2011 ◽  
Vol 11 (1) ◽  
pp. 30-41 ◽  
Author(s):  
Alejandro Sánchez-Arreguín ◽  
Ana Silvia Pérez-Martínez ◽  
Alfredo Herrera-Estrella
Keyword(s):  
Transcription Factors ◽  
Blue Light ◽  
Light Pulse ◽  
Mechanical Damage ◽  
Pathogenic Fungi ◽  
Mrna Levels ◽  
Trichoderma Atroviride ◽  
Proteomic Profiling ◽  
Content Type ◽  
Protein Levels

ABSTRACT The genus Trichoderma is one of the most widely used biological control agents of plant-pathogenic fungi. The main mechanism for survival and dispersal of Trichoderma is through the production of asexual spores (conidia). The transition from filamentous growth to conidiation can be triggered by light, nutrient deprivation, and mechanical damage of the mycelium. We conducted proteomic profiling analyses of Trichoderma atroviride after a blue light pulse. The use of two-dimensional electrophoresis (2-DE) and mass spectrometry (MS) analysis allowed us to identify 72 proteins whose expression was affected by blue light. Functional category analysis showed that the various proteins are involved in metabolism, cell rescue, and protein synthesis. We determined the relationship between mRNA levels of selected genes 30 min after a light pulse and protein expression levels at different times after the pulse and found this correlation to be very weak. The correlation was highest when protein and mRNA levels were compared for the same time point. The transcription factors BLR-1 and BLR-2 are vital to the photoconidiation process; here we demonstrate that both BLR proteins are active in darkness and affect several elements at both the transcript and protein levels. Unexpectedly, in darkness, downregulation of proteins prevailed in the Δ blr-1 mutant, while upregulation of proteins predominated in the Δ blr-2 mutant. Our data demonstrate that the BLR proteins play roles individually and as a complex.

scholarly journals Golgi UDP-GlcNAc:PolypeptideO-α-N-Acetyl-d-Glucosaminyltransferase 2 (TcOGNT2) Regulates Trypomastigote Production and Function in Trypanosoma cruzi

2014 ◽  
Vol 13 (10) ◽  
pp. 1312-1327 ◽  
Author(s):  
Carolina M. Koeller ◽  
Hanke van der Wel ◽  
Christa L. Feasley ◽  
Fernanda Abreu ◽  
Juliana Dutra Barbosa da Rocha ◽  
...  
Keyword(s):  
Cell Culture ◽  
Trypanosoma Cruzi ◽  
Low Frequency ◽  
Protozoan Parasite ◽  
Infection Frequency ◽  
Content Type ◽  
Protein Levels ◽  
Life Cycle Stages ◽  
Altered Cell ◽  
And Function

ABSTRACTAll life cycle stages of the protozoan parasiteTrypanosoma cruziare enveloped by mucin-like glycoproteins which, despite major changes in their polypeptide cores, are extensively and similarlyO-glycosylated.O-Glycan biosynthesis is initiated by the addition of αGlcNAc to Thr in a reaction catalyzed by Golgi UDP-GlcNAc:polypeptideO-α-N-acetyl-d-glucosaminyltransferases (ppαGlcNAcTs), which are encoded byTcOGNT1andTcOGNT2. We now directly show that TcOGNT2 is associated with the Golgi apparatus of the epimastigote stage and is markedly downregulated in both differentiated metacyclic trypomastigotes (MCTs) and cell culture-derived trypomastigotes (TCTs). The significance of downregulation was examined by forced continued expression of TcOGNT2, which resulted in a substantial increase of TcOGNT2 protein levels but only modestly increased ppαGlcNAcT activity in extracts and altered cell surface glycosylation in TCTs. Constitutive TcOGNT2 overexpression had no discernible effect on proliferating epimastigotes but negatively affected production of both types of trypomastigotes. MCTs differentiated from epimastigotes at a low frequency, though they were apparently normal based on morphological and biochemical criteria. However, these MCTs exhibited an impaired ability to produce amastigotes and TCTs in cell culture monolayers, most likely due to a reduced infection frequency. Remarkably, inhibition of MCT production did not depend on TcOGNT2 catalytic activity, whereas TCT production was inhibited only by active TcOGNT2. These findings indicate that TcOGNT2 downregulation is important for proper differentiation of MCTs and functioning of TCTs and that TcOGNT2 regulates these functions by using both catalytic and noncatalytic mechanisms.

Proteins Associated with Risk of Kidney Function Decline in the General Population

2021 ◽  
Vol 32 (9) ◽  
pp. 2291-2302
Author(s):  
Morgan E. Grams ◽  
Aditya Surapaneni ◽  
Jingsha Chen ◽  
Linda Zhou ◽  
Zhi Yu ◽  
...  
Keyword(s):  
Kidney Function ◽  
Plasma Proteins ◽  
Large Scale ◽  
Genome Wide Association Study ◽  
Meta Analysis ◽  
Proteomic Profiling ◽  
Content Type ◽  
Protein Levels ◽  
Time Periods ◽  
Egfr Decline

BackgroundProteomic profiling may allow identification of plasma proteins that associate with subsequent changesin kidney function, elucidating biologic processes underlying the development and progression of CKD.MethodsWe quantified the association between 4877 plasma proteins and a composite outcome of ESKD or decline in eGFR by ≥50% among 9406 participants in the Atherosclerosis Risk in Communities (ARIC) Study (visit 3; mean age, 60 years) who were followed for a median of 14.4 years. We performed separate analyses for these proteins in a subset of 4378 participants (visit 5), who were followed at a later time point, for a median of 4.4 years. For validation, we evaluated proteins with significant associations (false discovery rate <5%) in both time periods in 3249 participants in the Chronic Renal Insufficiency Cohort (CRIC) and 703 participants in the African American Study of Kidney Disease and Hypertension (AASK). We also compared the genetic determinants of protein levels with those from a meta-analysis genome-wide association study of eGFR.ResultsIn models adjusted for multiple covariates, including baseline eGFR and albuminuria, we identified 13 distinct proteins that were significantly associated with the composite end point in both time periods, including TNF receptor superfamily members 1A and 1B, trefoil factor 3, and β-trace protein. Of these proteins, 12 were also significantly associated in CRIC, and nine were significantly associated in AASK. Higher levels of each protein associated with higher risk of 50% eGFR decline or ESKD. We found genetic evidence for a causal role for one protein, lectin mannose-binding 2 protein (LMAN2).ConclusionsLarge-scale proteomic analysis identified both known and novel proteomic risk factors for eGFR decline.

scholarly journals Identification of the Haloarchaeal Phasin (PhaP) That Functions in Polyhydroxyalkanoate Accumulation and Granule Formation in Haloferax mediterranei

2012 ◽  
Vol 78 (6) ◽  
pp. 1946-1952 ◽  
Author(s):  
Shuangfeng Cai ◽  
Lei Cai ◽  
Hailong Liu ◽  
Xiaoqing Liu ◽  
Jing Han ◽  
...  
Keyword(s):  
Gene Knockout ◽  
Hypothetical Protein ◽  
Integrative Approach ◽  
Haloferax Mediterranei ◽  
Two Dimensional Gel Electrophoresis ◽  
Granule Formation ◽  
Content Type ◽  
Flight Mass Spectrometry ◽  
A Genome ◽  
Associated Proteins

ABSTRACTThe polyhydroxyalkanoate (PHA) granule-associated proteins (PGAPs) are important for PHA synthesis and granule formation, but currently little is known about the haloarchaeal PGAPs. This study focused on the identification and functional analysis of the PGAPs in the haloarchaeonHaloferax mediterranei. These PGAPs were visualized with two-dimensional gel electrophoresis (2-DE) and identified by matrix-assisted laser desorption ionization–tandem time of flight mass spectrometry (MALDI-TOF/TOF MS). The most abundant protein on the granules was identified as a hypothetical protein, designated PhaP. A genome-wide analysis revealed that thephaPgene is located upstream of the previously identifiedphaECgenes. Through an integrative approach of gene knockout/complementation and fermentation analyses, we demonstrated that this PhaP is involved in PHA accumulation. The ΔphaPmutant was defective in both PHA biosynthesis and cell growth compared to the wild-type strain. Additionally, transmission electron microscopy results indicated that the number of PHA granules in the ΔphaPmutant cells was significantly lower, and in most of the ΔphaPcells only a single large granule was observed. These results demonstrated that theH. mediterraneiPhaP was the predominant structure protein (phasin) on the PHA granules involved in PHA accumulation and granule formation. In addition, BLASTp and phylogenetic results indicate that this type of PhaP is exclusively conserved in haloarchaea, implying that it is a representative of the haloarchaeal type PHA phasin.

Conjugated Linoleic Acid Causes a Marked Increase in Liver α-Tocopherol and Liver α-Tocopherol Transfer Protein in C57BL/6 J Mice

2010 ◽  
Vol 80 (1) ◽  
pp. 65-73 ◽  
Author(s):  
Pei-Min Chao ◽  
Wan-Hsuan Chen ◽  
Chun-Huei Liao ◽  
Huey-Mei Shaw
Keyword(s):  
Antioxidant Enzymes ◽  
Linoleic Acid ◽  
Conjugated Linoleic Acid ◽  
Thiobarbituric Acid ◽  
Control Group ◽  
Thiobarbituric Acid Reactive Substances ◽  
Control Groups ◽  
Protein Levels ◽  
Transfer Protein ◽  
And Control

Conjugated linoleic acid (CLA) is a collective term for the positional and geometric isomers of a conjugated diene of linoleic acid (C18:2, n-6). The aims of the present study were to evaluate whether levels of hepatic α-tocopherol, α-tocopherol transfer protein (α-TTP), and antioxidant enzymes in mice were affected by a CLA-supplemented diet. C57BL/6 J mice were divided into the CLA and control groups, which were fed, respectively, a 5 % fat diet with or without 1 g/100 g of CLA (1:1 mixture of cis-9, trans-11 and trans-10, cis-12) for four weeks. α-Tocopherol levels in plasma and liver were significantly higher in the CLA group than in the control group. Liver α-TTP levels were also significantly increased in the CLA group, the α-TTP/β-actin ratio being 2.5-fold higher than that in control mice (p<0.01). Thiobarbituric acid-reactive substances were significantly decreased in the CLA group (p<0.01). There were no significant differences between the two groups in levels of three antioxidant enzymes (superoxide dismutase, glutathione peroxidase, and catalase). The accumulation of liver α-tocopherol seen with the CLA diet can be attributed to the antioxidant potential of CLA and the ability of α-TTP induction. The lack of changes in antioxidant enzyme protein levels and the reduced lipid peroxidation in the liver of CLA mice are due to α-tocopherol accumulation.

Pengaruh Lama Pengasinan Terhadap Kadar Protein Putih Telur Itik

2014 ◽  
Vol 1 (1) ◽  
pp. 36 ◽  
Author(s):  
Siti Fatimah ◽  
Muji Rahayu ◽  
Siti Aminah
Keyword(s):  
Protein Level ◽  
Egg White ◽  
Special Treatment ◽  
Egg White Protein ◽  
Protein Levels ◽  
Duck Egg ◽  
Egg Period ◽  
Graph Presentation ◽  
And Control ◽  
The Impact

Background : Egg  is one of the animal protein source, which has delicious taste, easy to digest and highly nutritious. Besides its affordable price, its supply availability is unquestionable as well. However, due to its short storability, it requires special treatment, such as preserving, to store it for long period. One way to preserve the egg is by pickling egg, which generally requires seven to ten days of marinating. During the process of marinating, there will be a visual change of egg white and yolk. Their structures  will be more solid (the occurrence of thickening process) because salinization will lead to protein denaturalization. Consequently, it has an influence as well towards the content of egg white protein of duck egg. This study is aimed to explore the impact of various time of pickling egg towards egg white protein of duck egg. Method  : The study where takes place in a laboratories, is a true experimental study for the reason that the researcher must provide intervention, hence all of potentially confounding variables are manageable. Samples that had been used in this study are duck eggs which were bought from North Brebes. This study is expected to generate data from four various time of pickling egg and control (no treatment). Since there are four samples, accordingly the number of data resulted are twenty. The resulted data will be descriptively presented in table, graph, presentation, and narration. Result  : Protein level examination within duck white egg shows changes  in protein levels that occurs in every variation of pickling egg time, where the average results of the assay of duck egg white protein is 14.94% without treatment (control), in five days of pickling time is 13.68%, in seven days of pickling time is 13.29%, in nine days of pickling time is 12.87% and eleven days of pickling time is 12.78%. Conclusion  : There is a significant impact among the period of pickling time to the protein level degradation of duck white egg. Keywords : Duck egg, period of pickling time, level protein of duck white egg.

Stereotactic radiation treatment planning and follow-up studies involving fused multimodality imaging

2004 ◽  
Vol 101 (Supplement3) ◽  
pp. 326-333 ◽  
Author(s):  
Klaus D. Hamm ◽  
Gunnar Surber ◽  
Michael Schmücking ◽  
Reinhard E. Wurm ◽  
Rene Aschenbach ◽  
...  
Keyword(s):  
Image Fusion ◽  
Treatment Planning ◽  
Radiation Treatment ◽  
Data Sets ◽  
Slice Thickness ◽  
Radiation Treatment Planning ◽  
Content Type ◽  
Follow Up Studies ◽  
Fine Print

Object. Innovative new software solutions may enable image fusion to produce the desired data superposition for precise target definition and follow-up studies in radiosurgery/stereotactic radiotherapy in patients with intracranial lesions. The aim is to integrate the anatomical and functional information completely into the radiation treatment planning and to achieve an exact comparison for follow-up examinations. Special conditions and advantages of BrainLAB's fully automatic image fusion system are evaluated and described for this purpose. Methods. In 458 patients, the radiation treatment planning and some follow-up studies were performed using an automatic image fusion technique involving the use of different imaging modalities. Each fusion was visually checked and corrected as necessary. The computerized tomography (CT) scans for radiation treatment planning (slice thickness 1.25 mm), as well as stereotactic angiography for arteriovenous malformations, were acquired using head fixation with stereotactic arc or, in the case of stereotactic radiotherapy, with a relocatable stereotactic mask. Different magnetic resonance (MR) imaging sequences (T1, T2, and fluid-attenuated inversion-recovery images) and positron emission tomography (PET) scans were obtained without head fixation. Fusion results and the effects on radiation treatment planning and follow-up studies were analyzed. The precision level of the results of the automatic fusion depended primarily on the image quality, especially the slice thickness and the field homogeneity when using MR images, as well as on patient movement during data acquisition. Fully automated image fusion of different MR, CT, and PET studies was performed for each patient. Only in a few cases was it necessary to correct the fusion manually after visual evaluation. These corrections were minor and did not materially affect treatment planning. High-quality fusion of thin slices of a region of interest with a complete head data set could be performed easily. The target volume for radiation treatment planning could be accurately delineated using multimodal information provided by CT, MR, angiography, and PET studies. The fusion of follow-up image data sets yielded results that could be successfully compared and quantitatively evaluated. Conclusions. Depending on the quality of the originally acquired image, automated image fusion can be a very valuable tool, allowing for fast (∼ 1–2 minute) and precise fusion of all relevant data sets. Fused multimodality imaging improves the target volume definition for radiation treatment planning. High-quality follow-up image data sets should be acquired for image fusion to provide exactly comparable slices and volumetric results that will contribute to quality contol.

Comparative Two-Dimensional Gel Electrophoresis of Trypanosoma cruzi Mammalian-Stage Forms in an Alkaline pH Range

2015 ◽  
Vol 22 (12) ◽  
pp. 1066-1075 ◽  
Author(s):  
Adriana Magalhães ◽  
Rayner Queiroz ◽  
Izabela Bastos ◽  
Jaime Santana ◽  
Marcelo Sousa ◽  
...  
Keyword(s):  
Trypanosoma Cruzi ◽  
Gel Electrophoresis ◽  
Two Dimensional ◽  
Alkaline Ph ◽  
Two Dimensional Gel Electrophoresis ◽  
Ph Range

Mean reversion in corporate leverage: evidence from India

2019 ◽  
Vol 45 (9) ◽  
pp. 1183-1198
Author(s):  
Gaurav S. Chauhan ◽  
Pradip Banerjee
Keyword(s):  
Capital Structure ◽  
Emerging Market ◽  
Simulated Data ◽  
Mean Reversion ◽  
Developed Countries ◽  
Data Sets ◽  
Debt Ratio ◽  
Testing Strategy ◽  
Content Type ◽  
Financing Behavior

Purpose Recent papers on target capital structure show that debt ratio seems to vary widely in space and time, implying that the functional specifications of target debt ratios are of little empirical use. Further, target behavior cannot be adjudged correctly using debt ratios, as they could revert due to mechanical reasons. The purpose of this paper is to develop an alternative testing strategy to test the target capital structure. Design/methodology/approach The authors make use of a major “shock” to the debt ratios as an event and think of a subsequent reversion as a movement toward a mean or target debt ratio. By doing this, the authors no longer need to identify target debt ratios as a function of firm-specific variables or any other rigid functional form. Findings Similar to the broad empirical evidence in developed economies, there is no perceptible and systematic mean reversion by Indian firms. However, unlike developed countries, proportionate usage of debt to finance firms’ marginal financing deficits is extensive; equity is used rather sparingly. Research limitations/implications The trade-off theory could be convincingly refuted at least for the emerging market of India. The paper here stimulated further research on finding reasons for specific financing behavior of emerging market firms. Practical implications The results show that the firms’ financing choices are not only depending on their own firm’s specific variables but also on the financial markets in which they operate. Originality/value This study attempts to assess mean reversion in debt ratios in a unique but reassuring manner. The results are confirmed by extensive calibration of the testing strategy using simulated data sets.

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