Studies of HLA, Factor B (Bf), Complement C2 and C4 Haplotypes in Type 1 Diabetic and Control Families from Northern Sweden

Abstract Objectives There are several hypotheses on the effects of the rs1738074 T/C single nucleotide polymorphism in the TAGAP gene; however, there has been no study on Turkish pediatric patients. We aimed to investigate the association of celiac disease (CD) and type 1 diabetes mellitus (T1DM) comorbidity with the polymorphism in the TAGAP gene of Turkish pediatric patients. Methods Totally, 127 pediatric CD patients and 100 healthy children were included. We determined the polymorphism by the allele-specific polymerase chain reaction method. We used IBM SPSS Statistics version 25.0 and Arlequin 3.5.2 for the statistical analyses. The authors have no conflict of interest. Results It was determined that 72% (n=154) of only CD patients had C allele, whereas 28% (n=60) had T allele. Of the patients with celiac and T1DM, 42.5% (n=17) and 57.5% (n=23) had T and C alleles, respectively. Of the individuals in control group, 67% (n=134) had C allele, whereas 33% (n=66) had T allele. Conclusions There was no significant difference in the genotype and allele frequencies between the patient and control groups (p>0.05). There was no significant association between the disease risk and the polymorphism in our study group.

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A/H1N1 hemagglutinin antibodies show comparable affinity in vaccine-related Narcolepsy type 1 and control and are unlikely to contribute to pathogenesis

Scientific Reports ◽

10.1038/s41598-021-83543-z ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

Alexander Lind ◽

Ilaria Marzinotto ◽

Cristina Brigatti ◽

Anita Ramelius ◽

Lorenzo Piemonti ◽

...

Keyword(s):

Antibody Response ◽

Antigenic Determinant ◽

Etiological Agent ◽

Healthy Controls ◽

Antibody Affinity ◽

Antibody Titres ◽

Antibody Levels ◽

And Control ◽

Radiobinding Assay

AbstractAn increased incidence of narcolepsy type 1 (NT1) was observed in Scandinavia following the 2009–2010 influenza Pandemrix vaccination. The association between NT1 and HLA-DQB1*06:02:01 supported the view of the vaccine as an etiological agent. A/H1N1 hemagglutinin (HA) is the main antigenic determinant of the host neutralization antibody response. Using two different immunoassays, the Luciferase Immunoprecipitation System (LIPS) and Radiobinding Assay (RBA), we investigated HA antibody levels and affinity in an exploratory and in a confirmatory cohort of Swedish NT1 patients and healthy controls vaccinated with Pandemrix. HA antibodies were increased in NT1 patients compared to controls in the exploratory (LIPS p = 0.0295, RBA p = 0.0369) but not in the confirmatory cohort (LIPS p = 0.55, RBA p = 0.625). HA antibody affinity, assessed by competition with Pandemrix vaccine, was comparable between patients and controls (LIPS: 48 vs. 39 ng/ml, p = 0.81; RBA: 472 vs. 491 ng/ml, p = 0.65). The LIPS assay also detected higher HA antibody titres as associated with HLA-DQB1*06:02:01 (p = 0.02). Our study shows that following Pandemrix vaccination, HA antibodies levels and affinity were comparable NT1 patients and controls and suggests that HA antibodies are unlikely to play a role in NT1 pathogenesis.

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OP0083 INFECTIOUS AND AUTOINFLAMMATORY MODIC TYPE 1 CHANGES HAVE DIFFERENT PATHOMECHANISMS

Annals of the Rheumatic Diseases ◽

10.1136/annrheumdis-2021-eular.825 ◽

2021 ◽

Vol 80 (Suppl 1) ◽

pp. 45.2-45

Author(s):

I. Heggli ◽

R. Schüpbach ◽

N. Herger ◽

T. A. Schweizer ◽

A. Juengel ◽

...

Keyword(s):

Flow Cytometry ◽

Immune Response ◽

T Cell ◽

Cell Populations ◽

Gene Set ◽

Go Enrichment ◽

Modic Type ◽

And Control ◽

Infectious Etiology

Background:Modic type 1 changes (MC1) are vertebral bone marrow (BM) edema that associate with non-specific low back pain (LBP). Two etiologies have been described. In the infectious etiology the anaerobic aerotolerant Cutibacterium acnes (C. acnes) invades damaged intervertebral discs (IVDs) resulting in disc infection and endplate damage, which leads to the evocation of an immune response. In the autoinflammatory etiology disc and endplate damage lead to the exposure of immune privileged disc cells and matrix to leukocytes, thereby evoking an immune response in the BM. Different etiologies require different treatment strategies. However, it is unknown if etiology-specific pathological mechanisms exist.Objectives:The aim of this study was to identify etiology-specific dysregulated pathways of MC1 and to perform in-depth analysis of immune cell populations of the autoinflammatory etiology.Methods:BM aspirates and biopsies were obtained from LBP patients with MC1 undergoing spinal fusion. Aspirates/biopsies were taken prior screw insertion through the pedicle screw trajectory. From each patient, a MC1 and an intra-patient control aspiration/biopsy from the adjacent vertebral level was taken. If C. acnes in IVDs adjacent to MC1 were detected by anaerobic bacterial culture, patients were assigned to the infectious, otherwise to the autoinflammatory etiology.Total RNA was isolated from aspirates and sequenced (Novaseq) (infectious n=3 + 3, autoinflammatory n=5 + 5). Genes were considered as differentially expressed (DEG) if p-value < 0.01 and log2fc > ± 0.5. Gene ontology (GO) enrichment was performed in R (GOseq), gene set enrichment analysis (GSEA) with GSEA software.Changes in cell populations of the autoinflammatory etiology were analyzed with single cell RNA sequencing (scRNAseq): Control and MC1 biopsies (n=1 + 1) were digested, CD45+CD66b- mononuclear cells isolated with fluorescence activated cell sorting (FACS), and 10000 cells were sequenced (10x Genomics). Seurat R toolkit was used for quality-control, clustering, and differential expression analysis.Transcriptomic changes (n=5 + 5) of CD45+CD66b+ neutrophils isolated with flow cytometry from aspirates were analyzed as for total bulk RNAseq. Neutrophil activation (n=3 + 3) was measured as CD66b+ expression with flow cytometry. CD66bhigh and CD66blow fractions in MC1 and control neutrophils were compared with paired t-test.Results:Comparing MC1 to control in total bulk RNAseq, 204 DEG in the autoinflammatory and 444 DEG in the infectious etiology were identified with only 67 shared genes (Fig. 1a). GO enrichment revealed “T-cell activation” (p = 2.50E-03) in the autoinflammatory and “complement activation, classical pathway” (p=1.1E-25) in the infectious etiology as top enriched upregulated biological processes (BP) (Fig 1b). ScRNAseq of autoinflammatory MC1 showed an overrepresentation of T-cells (p= 1.00E-34, OR=1.54) and myelocytes (neutrophil progenitor cells) (p=4.00E-05, OR=2.27) indicating an increased demand of these cells (Fig. 1c). Bulk RNAseq analysis of neutrophils from the autoinflammatory etiology revealed an activated, pro-inflammatory phenotype (Fig 1d), which was confirmed with more CD66bhigh neutrophils in MC1 (+11.13 ± 2.71%, p=0.02) (Fig. 1e).Figure 1.(a) Venn diagram of DEG from total bulk RNAseq (b) Top enriched upregulated BP of autoinflammatory (left) and infectious (right) etiology (c) Cell clustering of autoinflammatory MC1 BM (d) Enrichment of “inflammatory response” gene set in autoinflammatory MC1 neutrophils (e) Representative histogram of CD66b+ expression in MC1 and control neutrophils.Conclusion:Autoinflammatory and infectious etiologies of MC1 have different pathological mechanisms. T-cell and neutrophil activation seem to be important in the autoinflammatory etiology. This has clinical implication as it could be explored for diagnostic approaches to distinguish the two MC1 etiologies and supports developing targeted treatments for both etiologies.Disclosure of Interests:None declared

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Accounting for confounding by time, early intervention adoption, and time-varying effect modification in the design and analysis of stepped-wedge designs: application to a proposed study design to reduce opioid-related mortality

BMC Medical Research Methodology ◽

10.1186/s12874-021-01229-6 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Lior Rennert ◽

Moonseong Heo ◽

Alain H. Litwin ◽

Victor De Gruttola

Keyword(s):

Mixed Effects ◽

Effect Modification ◽

External Factors ◽

Mixed Effects Models ◽

Time Varying ◽

Intervention Effect ◽

Type 1 Error ◽

And Control ◽

Related Mortality

Abstract Background Beginning in 2019, stepped-wedge designs (SWDs) were being used in the investigation of interventions to reduce opioid-related deaths in communities across the United States. However, these interventions are competing with external factors such as newly initiated public policies limiting opioid prescriptions, media awareness campaigns, and the COVID-19 pandemic. Furthermore, control communities may prematurely adopt components of the intervention as they become available. The presence of time-varying external factors that impact study outcomes is a well-known limitation of SWDs; common approaches to adjusting for them make use of a mixed effects modeling framework. However, these models have several shortcomings when external factors differentially impact intervention and control clusters. Methods We discuss limitations of commonly used mixed effects models in the context of proposed SWDs to investigate interventions intended to reduce opioid-related mortality, and propose extensions of these models to address these limitations. We conduct an extensive simulation study of anticipated data from SWD trials targeting the current opioid epidemic in order to examine the performance of these models in the presence of external factors. We consider confounding by time, premature adoption of intervention components, and time-varying effect modification— in which external factors differentially impact intervention and control clusters. Results In the presence of confounding by time, commonly used mixed effects models yield unbiased intervention effect estimates, but can have inflated Type 1 error and result in under coverage of confidence intervals. These models yield biased intervention effect estimates when premature intervention adoption or effect modification are present. In such scenarios, models incorporating fixed intervention-by-time interactions with an unstructured covariance for intervention-by-cluster-by-time random effects result in unbiased intervention effect estimates, reach nominal confidence interval coverage, and preserve Type 1 error. Conclusions Mixed effects models can adjust for different combinations of external factors through correct specification of fixed and random time effects. Since model choice has considerable impact on validity of results and study power, careful consideration must be given to how these external factors impact study endpoints and what estimands are most appropriate in the presence of such factors.

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Comparison of Measurements of Autoantibodies to Glutamic Acid Decarboxylase and Islet Antigen-2 in Whole Blood Eluates from Dried Blood Spots Using the RSR-Enzyme Linked Immunosorbent Assay Kits and In-House Radioimmunoassays

Experimental Diabetes Research ◽

10.1155/2010/173652 ◽

2010 ◽

Vol 2010 ◽

pp. 1-6 ◽

Cited By ~ 4

Author(s):

Anders Persson ◽

Charlotte Becker ◽

Ida Hansson ◽

Anita Nilsson ◽

Carina Törn

Keyword(s):

Glutamic Acid ◽

Glutamic Acid Decarboxylase ◽

Dried Blood Spots ◽

Enzyme Linked Immunosorbent Assay ◽

Acid Decarboxylase ◽

Blood Spots ◽

And Control ◽

Islet Antigen ◽

Prozone Effect

To evaluate the performance of dried blood spots (DBSs) with subsequent analyses of glutamic acid decarboxylase (GADA) and islet antigen-2 (IA-2A) with the RSR-ELISAs, we selected 80 children newly diagnosed with type 1 diabetes and 120 healthy women. DBSs from patients and controls were used for RSR-ELISAs while patients samples were analysed also with in-house RIAs. The RSR-ELISA-GADA performed well with a specificity of 100%, albeit sensitivity (46%) was lower compared to in RIA (56%;P=.008). No prozone effect was observed after dilution of discrepant samples. RSR-ELISA-IA-2A achieved specificity of 69% and sensitivity was lower (59%) compared with RIA (66%;P<.001). Negative or low positive patients and control samples in the RSR-ELISA-IA-2A increased after dilution. Eluates from DBS can readily be used to analyse GADA with the RSR-ELISA, even if low levels of autoantibodies were not detected. Some factor could disturb RSR-ELISA-IA-2A analyses.

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Differences in splicing defects between the grey and white matter in myotonic dystrophy type 1

10.1101/819433 ◽

2019 ◽

Author(s):

Masamitsu Nishi ◽

Takashi Kimura ◽

Mitsuru Furuta ◽

Koichi Suenaga ◽

Tsuyoshi Matsumura ◽

...

Keyword(s):

White Matter ◽

Myotonic Dystrophy ◽

Cell Body ◽

Neuronal Cell ◽

Neuronal Cell Body ◽

Myotonic Dystrophy Type 1 ◽

Myotonic Dystrophy Type ◽

And Control ◽

The Brain

AbstractMyotonic dystrophy type 1 (DM1) is a multi-system disorder caused by CTG repeats in the myotonic dystrophy protein kinase (DMPK) gene. This leads to sequestration of the splicing factor, muscleblind-like 2 (MBNL2), and aberrant splicing, mainly in the central nervous system. We investigated the splicing patterns of MBNL1/2 and genes controlled by MBNL2 in several regions of the brain and between the grey matter (GM) and white matter (WM) in DM1 patients using RT-PCR. Compared with the control, the percentage of spliced-in parameter (PSI) for most of the examined exons were significantly altered in most of the brain regions of DM1 patients, except for the cerebellum. The splicing of many genes was differently regulated between the GM and WM in both DM1 and control. The level of change in PSI between DM1 and control was higher in the GM than in the WM. The differences in alternative splicing between the GM and WM may be related to the effect of DM1 on the WM of the brain. We hypothesize that in DM1, aberrantly spliced isoforms in the neuronal cell body of the GM may not be transported to the axon. This might affect the WM as a consequence of Wallerian degeneration secondary to cell body damage. Our findings may have implications for analysis of the pathological mechanisms and exploring potential therapeutic targets.

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Integrated management of tough lovegrass (Eragrostis plana Nees): associating chemical control tools and plant physiology

Ciência Rural ◽

10.1590/0103-8478cr20200271 ◽

2021 ◽

Vol 51 (2) ◽

Author(s):

Eduardo Avelino Faleiro ◽

Diego Martins Chiapinotto ◽

Fabiane Pinto Lamego ◽

Carlos Eduardo Schaedler ◽

Eduardo Bohrer de Azevedo

Keyword(s):

Solar Radiation ◽

Integrated Management ◽

Dry Mass ◽

Radiation Factor ◽

Factor B ◽

Randomized Design ◽

Completely Randomized Design ◽

Factor C ◽

Resource Conditions ◽

And Control

ABSTRACT: The objective of the study was to evaluate the efficiency of herbicides use in the control of tough lovegrass according to the availability of solar radiation and the presence or absence of flooded. Two experiments were conducted in a completely randomized design, in a 2x2x4 and 2x2x5 factorial scheme, where factor A was equivalent to the environment (natural or reduced radiation); factor B to water condition (with a 2 cm flooded or without); and, factor C the herbicides: cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), cyhalofop butyl + glyphosate (315 g ha-1 + 1080 g ha-1) and control (without application) in the first experiment. In the second experiment, the herbicides used were cyhalofop butyl (315 g ha-1), glyphosate (1080 g ha-1), sethoxydim (184g ha-1), imazethapyr (106 g ha-1) and control (without application). Glyphosate efficiently controls plants of tough lovegrass with four tillers (>90%), being superior to cyhalofop, imazethapyr and sethoxydim; independently, of resource conditions. Reduction in the availability of solar radiation generates less shoot dry mass production from the weed, and improves the control only by imazethapyr and cyhalofop. In general, a flooded condition does not affect tough lovegrass control by herbicides.

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Effect of Black Grape Seed(Vitis Vinifera) Extract Consumption with Moderate-Intensity Aerobic Training on the Expression of MicroRNAs in Type 1 Diabetic Cardiac Tissue Rats

Iranian journal of diabetes and obesity ◽

10.18502/ijdo.v13i4.8003 ◽

2021 ◽

Author(s):

Maryam Shirani Bidabadi ◽

Jamshid Banaei Borojeni ◽

Saeed Keshavarz ◽

Mohammad Karimi

Keyword(s):

Cardiac Tissue ◽

Aerobic Training ◽

Grape Seed ◽

Diabetic Control ◽

Seed Extract ◽

Moderate Intensity ◽

P Value ◽

The Difference ◽

And Control

Objective: This study aimed to evaluate the effect of consuming grape seed extract with moderate-intensity aerobic training on the expression of miR-126 and miR-29 in the cardiac tissue in type 1 diabetic male rats. Materials and Methods: 40 rats with an initial weight range of 160-220 g were divided into five groups: Training + Extract, Training, Extract, Diabetic / Control, and Healthy / Control. Aerobic training program was moderate intensity and rats performed aerobic training for 60 minutes a day with the intensity 70 to 75% of maximum oxygen consumption (28 meters per minute). Grape seed extract was also administered by gavage at a dose of 40 mg/kg per day. Results: Expression of both miRNAs in the three groups of training + extract, healthy training and control was significantly higher than the two groups of extract and diabetic control (P-value= 0.001). The difference between the three groups of training + extract, healthy training and control and also the difference between the two groups of extract and diabetic control were not significant (P-value> 0.05). Conclusion: Aerobic training may be able to prevent cardiac disease caused by type 1 diabetes.

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