160 research outputs found
Anti-α-enolase Antibodies in Serum from Pediatric Patients Affected by Inflammatory Diseases: Diagnostic and Pathogenetic Insights
Human glycolytic enzyme α-enolase was associated with human diseases and with inflammation. An ELISA test was developed to measure anti-α-enolase AAE IgG and AAE IgA in the serum from patients affected by inflammatory diseases with the purpose to evaluate it as a novel diagnostic marker. 80 healthy blood donors and 194 paediatric patients affected by Juvenile idiopathic arthritis (JIA), celiac disease (CD), Crohn's Disease (CrD), hereditary periodic fever (HPF), and PFAPA syndrome were included in the study. HPF patients showed high levels of AAE antibodies, whereas JIA, CD, and CrD presented only partial results. Benign fevers such as PFAPA were almost negative for AAE Abs. These findings suggested that the genetic dysfunction of inflammasome associated with HPF could lead to the formation of AAE Abs that could be used for an early and easy diagnosis
Evaluation of Proton-Induced Biomolecular Changes in MCF-10A Breast Cells by Means of FT-IR Microspectroscopy
Radiotherapy (RT) with accelerated beams of charged particles (protons and carbon ions),
also known as hadrontherapy, is a treatment modality that is increasingly being adopted thanks to the
several benefits that it grants compared to conventional radiotherapy (CRT) treatments performed
by means of high-energy photons/electrons. Hence, information about the biomolecular effects in
exposed cells caused by such particles is needed to better realize the underlying radiobiological
mechanisms and to improve this therapeutic strategy. To this end, Fourier transform infrared
microspectroscopy (-FT-IR) can be usefully employed, in addition to long-established radiobiological
techniques, since it is currently considered a helpful tool for examining radiation-induced cellular
changes. In the present study, MCF-10A breast cells were chosen to evaluate the effects of proton
exposure using -FT-IR. They were exposed to different proton doses and fixed at various times
after exposure to evaluate direct effects due to proton exposure and the kinetics of DNA damage
repair. Irradiated and control cells were examined in transflection mode using low-e substrates that
have been recently demonstrated to offer a fast and direct way to examine proton-exposed cells. The
acquired spectra were analyzed using a deconvolution procedure and a ratiometric approach, both of
which showed the different contributions of DNA, protein, lipid, and carbohydrate cell components.
These changes were particularly significant for cells fixed 48 and 72 h after exposure. Lipid changes
were related to variations in membrane fluidity, and evidence of DNA damage was highlighted.
The analysis of the Amide III band also indicated changes that could be related to different enzyme
contributions in DNA repair
FT-IR Transflection Micro-Spectroscopy Study on Normal Human Breast Cells after Exposure to a Proton Beam
Fourier transform infrared micro-spectroscopy (mu-FT-IR) is nowadays considered a valuable tool for investigating the changes occurring in human cells after exposure to ionizing radiation. Recently, considerable attention has been devoted to the use of this optical technique in the study of cells exposed to proton beams, that are being increasingly adopted in cancer therapy. Different experimental configurations are used for proton irradiation and subsequent spectra acquisition. To facilitate the use of mu-FT-IR, it may be useful to investigate new experimental approaches capable of speeding up and simplifying the irradiation and measurements phases. Here, we propose the use of low-e-substrates slides for cell culture, allowing the irradiation and spectra acquisition in transflection mode in a fast and direct way. In recent years, there has been a wide debate about the validity of these supports, but many researchers agree that the artifacts due to the presence of the electromagnetic standing wave effects are negligible in many practical cases. We investigated human normal breast cells (MCF-10 cell line) fixed immediately after the irradiation with graded proton radiation doses (0, 0.5, 2, and 4 Gy). The spectra obtained in transflection geometry showed characteristics very similar to those present in the spectra acquired in transmission geometry and confirm the validity of the chosen approach. The analysis of spectra indicates the occurrence of significant changes in DNA and lipids components of cells. Modifications in protein secondary structure are also evidenced
Mitochondrial Dysfunction in Cancer and Neurodegenerative Diseases: Spotlight on Fatty Acid Oxidation and Lipoperoxidation Products
In several human diseases, such as cancer and neurodegenerative diseases, the levels of reactive oxygen species (ROS), produced mainly by mitochondrial oxidative phosphorylation, is increased. In cancer cells, the increase of ROS production has been associated with mtDNA mutations that, in turn, seem to be functional in the alterations of the bioenergetics and the biosynthetic state of cancer cells. Moreover, ROS overproduction can enhance the peroxidation of fatty acids in mitochondrial membranes. In particular, the peroxidation of mitochondrial phospholipid cardiolipin leads to the formation of reactive aldehydes, such as 4-hydroxynonenal (HNE) and malondialdehyde (MDA), which are able to react with proteins and DNA. Covalent modifications of mitochondrial proteins by the products of lipid peroxidation (LPO) in the course of oxidative cell stress are involved in the mitochondrial dysfunctions observed in cancer and neurodegenerative diseases. Such modifications appear to affect negatively mitochondrial integrity and function, in particular energy metabolism, adenosine triphosphate (ATP) production, antioxidant defenses and stress responses. In neurodegenerative diseases, indirect confirmation for the pathogenetic relevance of LPO-dependent modifications of mitochondrial proteins comes from the disease phenotypes associated with their genetic alterations
Clinical effectiveness of different natalizumab interval dosing schedules in a large Italian population of patients with multiple sclerosis
Introduction Natalizumab (NTZ) is one of the most
effective treatment options for multiple sclerosis (MS)
treatment. Our study aimed to evaluate the effectiveness
of NTZ when administered according to the extended
dosing strategy compared with standard 4-weekly
administration in a large Italian MS population.
Materials and methods This retrospective multicentre
study included patients with relapsing-remitting
MS
(RR-MS)
who received NTZ administrations between the
1 June 2012 and the 15 May 2018 and were followed
by the ’Italian MS Register’. All patients with MS were
stratified into two groups based on NTZ administration
schedule: standard interval dosing (SID) patients who
received infusions on average from 28 to 32 days
(median 30) and extended interval dosing (EID) including
patients who have been infused with interval between
33 and 49 days (median 43). Clinical data were assessed
at baseline (before starting NTZ), after 12 (T1) and 24
months (T2) of treatment.
Results Out of 5231 patients with RR-MS
screened,
2092 (mean age 43.2±12.0, 60.6% women) were
enrolled. A total of 1254 (59.9%) received NTZ
according to SID, and 838 (40.1%) according to EID. At
12 and 24 months, no differences in terms of annualised
relapse rate and disability status were found between the
two groups. Progression index and confirmed disability
worsening were similar between the two groups.
Discussion The use of NTZ with an extended interval
schedule showed similar effectiveness compared with
SID. Unchanged clinical efficacy of EID schedule may
raise the question of a possible advantage in terms of
tolerability and safety
Improving the accuracy of the structure prediction of the third hypervariable loop of the heavy chains of antibodies
Motivation: Antibodies are able to recognize a wide range of antigens through their complementary determining regions formed by six hypervariable loops. Predicting the 3D structure of these loops is essential for the analysis and reengineering of novel antibodies with enhanced affinity and specificity. The canonical structure model allows high accuracy prediction for five of the loops. The third loop of the heavy chain, H3, is the hardest to predict because of its diversity in structure, length and sequence composition.Results: We describe a method, based on the Random Forest automatic learning technique, to select structural templates for H3 loops among a dataset of candidates. These can be used to predict the structure of the loop with a higher accuracy than that achieved by any of the presently available methods. The method also has the advantage of being extremely fast and returning a reliable estimate of the model quality.Availability and implementation: The source code is freely available at http://www.biocomputing.it/H3Loopred/Contact: [email protected] Information: Supplementary data are available at Bioinformatics online
Convolutional neural network based on fluorescein angiography images for retinopathy of prematurity management
Purpose: The purpose of this study was to explore the use of fluorescein angiography (FA) images in a convolutional neural network (CNN) in the management of retinopathy of prematurity (ROP).Methods: The dataset involved a total of 835 FA images of 149 eyes (90 patients), where each eye was associated with a binary outcome (57 "untreated" eyes and 92 "treated"; 308 "untreated" images, 527 "treated"). The resolution of the images was 1600 and 1200 px in 20% of cases, whereas the remaining 80% had a resolution of 640 and 480 px. All the images were resized to 640 and 480 px before training and no other preprocessing was applied. A CNN with four convolutional layers was trained on 90% of the images (n = 752) randomly chosen. The accuracy of the prediction was assessed on the remaining 10% of images (n = 83). Keras version 2.2.0 for R with Tensorflow backend version 1.11.0 was used for the analysis.Results: The validation accuracy after 100 epochs was 0.88, whereas training accuracy was 0.97. The receiver operating characteristic (ROC) presented an area under the curve (AUC) of 0.91.Conclusions: Our study showed, we believe for the first time, the applicability of artificial intelligence (CNN) technology in the ROP management driven by FA. Further studies are needed to exploit different fields of applications of this technology.Translational Relevance: This algorithm is the basis for a system that could be applied to both ROP as well as experimental oxygen induced retinopathy
Risk of Getting COVID-19 in People With Multiple Sclerosis
Background and Objectives
Several studies have assessed risk factors associated with the severity of COVID-19 outcomes in
people with multiple sclerosis (PwMS). The potential role of disease-modifying therapies
(DMTs) and demographic and clinical factors on the risk of acquiring SARS-CoV-2 infection
has not been evaluated so far. The objective of this study was to assess risk factors of contracting
SARS-CoV-2 infection in PwMS by using data collected in the Italian MS Register (IMSR).
Methods
Acase-control (1:2) studywas set up. Cases included PwMSwith a confirmed diagnosis ofCOVID-19,
and controls included PwMS without a confirmed diagnosis of COVID-19. Both groups were propensity
score–matched by the date of COVID-19 diagnosis, the date of last visit, and the region of
residence. No healthy controls were included in this study. COVID-19 risk was estimated by multivariable
logistic regression models including demographic and clinical covariates. The impact of DMTs
was assessed in 3 independent logistic regression models including one of the following covariates: last
administeredDMT, previousDMTsequences, or the place where the last treatment was administered.
Results
A total of 779 PwMS with confirmed COVID-19 (cases) were matched to 1,558 PwMS without
COVID-19 (controls). In all 3 models, comorbidities, female sex, and a younger age were significantly
associated (p < 0.02)with a higher risk of contractingCOVID-19. Patients receiving natalizumab as last
DMT(OR[95%CI]: 2.38 [1.66–3.42], p < 0.0001) and those who underwent an escalation treatment
strategy (1.57 [1.16–2.13], p = 0.003) were at significantly higher COVID-19 risk. Moreover, PwMS receiving their last DMT requiring
hospital access (1.65 [1.34–2.04], p < 0.0001) showed a significant higher risk than those taking self-administered DMTs at home.
Discussion
This case-control study embedded in the IMSR showed that PwMS at higher COVID-19 risk are younger, more frequently
female individuals, and with comorbidities. Long-lasting escalation approach and last therapies that expose patients to the
hospital environment seem to significantly increase the risk of SARS-CoV2 infection in PwMS.
Classification of Evidence
This study provides Class III evidence that among patients with MS, younger age, being female individuals, having more
comorbidities, receiving natalizumab, undergoing an escalating treatment strategy, or receiving treatment at a hospital were
associated with being infected with COVID-19. Among patients with MS who were infected with COVID-19, a severe course
was associated with increasing age and having a progressive form of MS, whereas not being on treatment or receiving an
interferon beta agent was protective
Albiglutide, a Long Lasting Glucagon-Like Peptide-1 Analog, Protects the Rat Heart against Ischemia/Reperfusion Injury: Evidence for Improving Cardiac Metabolic Efficiency
BACKGROUND: The cardioprotective effects of glucagon-like peptide-1 (GLP-1) and analogs have been previously reported. We tested the hypothesis that albiglutide, a novel long half-life analog of GLP-1, may protect the heart against I/R injury by increasing carbohydrate utilization and improving cardiac energetic efficiency. METHODS/PRINCIPAL FINDINGS: Sprague-Dawley rats were treated with albiglutide and subjected to 30 min myocardial ischemia followed by 24 h reperfusion. Left ventricle infarct size, hemodynamics, function and energetics were determined. In addition, cardiac glucose disposal, carbohydrate metabolism and metabolic gene expression were assessed. Albiglutide significantly reduced infarct size and concomitantly improved post-ischemic hemodynamics, cardiac function and energetic parameters. Albiglutide markedly increased both in vivo and ex vivo cardiac glucose uptake while reducing lactate efflux. Analysis of metabolic substrate utilization directly in the heart showed that albiglutide increased the relative carbohydrate versus fat oxidation which in part was due to an increase in both glucose and lactate oxidation. Metabolic gene expression analysis indicated upregulation of key glucose metabolism genes in the non-ischemic myocardium by albiglutide. CONCLUSION/SIGNIFICANCE: Albiglutide reduced myocardial infarct size and improved cardiac function and energetics following myocardial I/R injury. The observed benefits were associated with enhanced myocardial glucose uptake and a shift toward a more energetically favorable substrate metabolism by increasing both glucose and lactate oxidation. These findings suggest that albiglutide may have direct therapeutic potential for improving cardiac energetics and function
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