Distinct transthyretin oxidation isoform profile in spinal fluid from patients with Alzheimer’s disease and mild cognitive impairment

BACKGROUND: Transthyretin (TTR), an abundant protein in cerebrospinal fluid (CSF), contains a free, oxidation-prone cysteine residue that gives rise to TTR isoforms. These isoforms may reflect conditions in vivo. Since increased oxidative stress has been linked to neurodegenerative disorders such as Alzheimer’s disease (AD) it is of interest to characterize CSF-TTR isoform distribution in AD patients and controls. Here, TTR isoforms are profiled directly from CSF by an optimized immunoaffinity-mass spectrometry method in 76 samples from patients with AD (n = 37), mild cognitive impairment (MCI, n = 17)), and normal pressure hydrocephalus (NPH, n = 15), as well as healthy controls (HC, n = 7). Fractions of three specific oxidative modifications (S-cysteinylation, S-cysteinylglycinylation, and S-glutathionylation) were quantitated relative to the total TTR protein. Results were correlated with diagnostic information and with levels of CSF AD biomarkers tau, phosphorylated tau, and amyloid β(1-42) peptide. RESULTS: Preliminary data highlighted the high risk of artifactual TTR modification due to ex vivo oxidation and thus the samples for this study were all collected using strict and uniform guidelines. The results show that TTR is significantly more modified on Cys(10) in the AD and MCI groups than in controls (NPH and HC) (p ≤ 0.0012). Furthermore, the NPH group, while having normal TTR isoform distribution, had significantly decreased amyloid β peptide but normal tau values. No obvious correlations between levels of routine CSF biomarkers for AD and the degree of TTR modification were found. CONCLUSIONS: AD and MCI patients display a significantly higher fraction of oxidatively modified TTR in CSF than the control groups of NPH patients and HC. Quantitation of CSF-TTR isoforms thus may provide diagnostic information in patients with dementia symptoms but this should be explored in larger studies including prospective studies of MCI patients. The development of methods for simple, robust, and reproducible inhibition of in vitro oxidation during CSF sampling and sample handling is highly warranted. In addition to the diagnostic information the possibility of using TTR as a CSF oxymeter is of potential value in studies monitoring disease activity and developing new drugs for neurodegenerative diseases

Specific autoantibody profiles and disease subgroups correlate with circulating micro-RNA in systemic sclerosis.

To evaluate the expression profiles of cell-free plasma miRNAs in SSc and to characterize their correlation with disease subgroups (lcSSc and dcSSc) and with autoantibody profiles

Profiling microRNAs in lung tissue from pigs infected with Actinobacillus pleuropneumoniae

<p>Abstract</p> <p>Background</p> <p>MicroRNAs (miRNAs) are a class of non-protein-coding genes that play a crucial regulatory role in mammalian development and disease. Whereas a large number of miRNAs have been annotated at the structural level during the latest years, functional annotation is sparse. <it>Actinobacillus pleuropneumoniae</it> (APP) causes serious lung infections in pigs. Severe damage to the lungs, in many cases deadly, is caused by toxins released by the bacterium and to some degree by host mediated tissue damage. However, understanding of the role of microRNAs in the course of this infectious disease in porcine is still very limited.</p> <p>Results</p> <p>In this study, the RNA extracted from visually unaffected and necrotic tissue from pigs infected with <it>Actinobacillus pleuropneumoniae</it> was subjected to small RNA deep sequencing. We identified 169 conserved and 11 candidate novel microRNAs in the pig. Of these, 17 were significantly up-regulated in the necrotic sample and 12 were down-regulated. The expression analysis of a number of candidates revealed microRNAs of potential importance in the innate immune response. MiR-155, a known key player in inflammation, was found expressed in both samples. Moreover, miR-664-5p, miR-451 and miR-15a appear as very promising candidates for microRNAs involved in response to pathogen infection.</p> <p>Conclusions</p> <p>This is the first study revealing significant differences in composition and expression profiles of miRNAs in lungs infected with a bacterial pathogen. Our results extend annotation of microRNA in pig and provide insight into the role of a number of microRNAs in regulation of bacteria induced immune and inflammatory response in porcine lung.</p

Mass-Spectrometry Based Proteome Comparison of Extracellular Vesicle Isolation Methods:Comparison of ME-kit, Size-Exclusion Chromatography, and High-Speed Centrifugation

Extracellular vesicles (EVs) are small membrane-enclosed particles released by cells under various conditions specific to cells&rsquo; biological states. Hence, mass-spectrometry (MS) based proteome analysis of EVs in plasma has gained much attention as a method to discover novel protein biomarkers. MS analysis of EVs in plasma is challenging and EV isolation is usually necessary. Therefore, we compared differences in abundance, subtypes, and contamination for EVs isolated by high-speed centrifugation, size exclusion chromatography (SEC), and peptide-affinity precipitation (PAP/ME kit) for subsequent MS-based proteome analysis. Successful EV isolation was evaluated by nanoparticle-tracking analysis, immunoblotting, and transmission electron microscopy, while EV abundance, EV subtypes, and contamination was evaluated by label-free tandem MS. High-speed centrifugation and SEC isolates showed high EV abundance at the expense of contamination by non-EV proteins and lipoproteins, respectively. These two methods also resulted in EVs of a similar type, however, with smaller EVs in SEC isolates. PAP isolates had a relatively low EV abundance and high contamination. We consider high-speed centrifugation and SEC suitable as EV isolation for MS biomarker studies, where the choice between the two should depend on the scientific questions and whether the focus is on larger or smaller EVs or a combination of both

Propuesta de mejora en el área de producción para reducir costos operativos en la empresa Inca Verde del Perú SAC

RESUMEN El presente trabajo tuvo como objetivo general: Reducir los costos operativos de la empresa Inca Verde del Perú S.A.C., a través de la propuesta de mejora en el área de producción. Realizando un diagnóstico de la situación actual con el uso del análisis causa-efecto o también llamado Diagrama de Ishikawa para identificar las principales causas del problema las cuales principalmente fueron: Deficiente planificación de la producción, falta de estudio de tiempos y movimientos, inexistencia de un plan de capacitación en los principales procesos de producción entre otros. Para los cuales se propusieron herramientas de mejora tales como: Plan de Requerimiento de Materiales, Estudio de Tiempos, Plan de capacitación, Aplicación de 5 S entre otros. Los resultados que se lograron fueron una reducción de costos en S/148.837 soles anuales de un total inicial de S/. S/. 315.471,53 Finalmente se realizó la evaluación económica financiera para la propuesta de mejora en el área de producción, obteniendo un VAN de S/. 128,567, un TIR de 80% y un Beneficio Costo de 1.41 lo cual indica la viabilidad del proyecto.ABSTRACT The present work had as general objective: Reduces the operative costs of the business Inca Verde del f Peru S.A.C., trought the proffer of improvement in the area of production. Realizing a diagnosis of the current situation with the use of the analysis cause - effect or also called Ishikawa's Graph to identify the principal reasons of the problem, which principally were: Deficient planning of the production, lack of study of times and movements, non-existence of a plan of training in the principal processes of production and others. For which, proposed tools of improvement such as: Plan of Requirement of Materials, Study of Times, Plan of training, Application of 5 S between others The results achieve wee The results achieved were a reduction of costs from S/148.837 annual to S/. S/. 315.471,53. annual Finally, the economic financial evaluation for the propuse of improvement in the area of production, obtaining a VNA of S/. 128,567, one TIR of 80 % and a Benefit Cost of 1.41 which indicates the viability of the project

Large Gliadin Peptides Detected in the Pancreas of NOD and Healthy Mice following Oral Administration

Gluten promotes type 1 diabetes in nonobese diabetic (NOD) mice and likely also in humans. In NOD mice and in non-diabetes-prone mice, it induces inflammation in the pancreatic lymph nodes, suggesting that gluten can initiate inflammation locally. Further, gliadin fragments stimulate insulin secretion from beta cells directly. We hypothesized that gluten fragments may cross the intestinal barrier to be distributed to organs other than the gut. If present in pancreas, gliadin could interact directly with the immune system and the beta cells to initiate diabetes development. We orally and intravenously administered 33-mer and 19-mer gliadin peptide to NOD, BALB/c, and C57BL/6 mice and found that the peptides readily crossed the intestinal barrier in all strains. Several degradation products were found in the pancreas by mass spectroscopy. Notably, the exocrine pancreas incorporated large amounts of radioactive label shortly after administration of the peptides. The study demonstrates that, even in normal animals, large gliadin fragments can reach the pancreas. If applicable to humans, the increased gut permeability in prediabetes and type 1 diabetes patients could expose beta cells directly to gliadin fragments. Here they could initiate inflammation and induce beta cell stress and thus contribute to the development of type 1 diabetes

The Urine Proteome Profile Is Different in Neuromyelitis Optica Compared to Multiple Sclerosis: a Clinical Proteome Study

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Circulating Antinuclear Antibodies in Patients with Pelvic Masses Are Associated with Malignancy and Decreased Survival

BACKGROUND: Circulating autoantibodies occur more frequently in cancer patients than in patients without cancer. METHODS AND FINDINGS: We examined sera from patients referred for pelvic mass symptoms to a tertiary university clinic. A total of 127 were diagnosed with epithelial ovarian cancer while 386 had a benign condition. A screen for IgG anti-nuclear antibodies (ANA) by indirect immunofluorescence on HEp-2 cells confirmed a highly significant overrepresentation of ANA in the cancer group where 40% had detectable (i.e., a titer ≥160) ANA compared with less than 12% in the benign group. The overrepresentation of ANA in the cancer group persisted (p<0.0001) after matching the age-profile of the benign group with the ovarian cancer group. Only 19 out of 127 patients in the age-matched benign subgroup were positive for ANA corresponding to an 85% specificity at 40% sensitivity of ANA as the only marker for malignancy. No correlation of ANA positivity in either group with specific bands in immunoblots could be demonstrated even though immunoblot positivity was clearly increased in the malignant group (41% vs. 3%). The presence, strength, and type of ANA did not correlate with serum CA-125 values or with staging, and ANA outcome did not contribute with independent diagnostic information. However, survival was significantly shorter in ANA-positive compared with ANA-negative cancer patients and patients with CA-125 below the median CA-125 value in the cancer group had a significantly decreased survival when positive for ANA. ANA status made no difference in the group with CA-125 values above the median. Also, there was a significant correlation between speckled ANA-strength and histological tumor grade. CONCLUSIONS: Circulating antibodies are a promising source for new biomarkers in cancer. Characterization of epitope specificities and measurements of consecutive samples will be important for further elucidating the role of ANA in evaluating ovarian cancer patients