Tear miRNAs identified in a murine model of Sjögren′s Syndrome as potential diagnostic biomarkers and indicators of disease mechanism

Objective: The tear miRNAome of the male NOD mouse, a model of ocular symptoms of Sjögren′s syndrome (SS), was analyzed to identify possible tear biomarkers of the disease. Methods: Male NOD mice, aged 12-14 weeks, were used to identify tear miRNAs associated with development of autoimmune dacryoadenitis. Age-matched female NOD mice that do not develop the autoimmune dacryoadenitis characteristic of SS were used as negative controls while age- and sex-matched male BALB/c mice served as healthy controls. Total RNA was isolated from stimulated tears pooled from 5 mice per sample and tear miRNAs were sequenced and analyzed. Putative miRNA hits were validated using RT-qPCR in a separate mouse cohort, and the pathways influenced by the validated hits were identified using Ingenuity Pathway Analysis. Results: In comparison to tears from both healthy (male BALB/c) and negative control (female NOD) mice, initial analysis identified 7 upregulated and 7 downregulated miRNAs in male NOD mouse tears. Of these, 8 were subsequently validated by RT-qPCR in tears from additional mouse cohorts. miRNAs previously implicated in SS pathology included mmu-miR-146a/b-5p, which were significantly downregulated in the male NOD mouse tears, as well as mmu-miR-150-5p and mmu-miR-181a-5p, which were upregulated in the male NOD mouse tears. All other validated hits including the upregulated miR-181b-5p and mmu-miR-203-3p, as well as the downregulated mmu-miR-322-5p and mmu-miR-503-5p, represent novel putative indicators of autoimmune dacryoadenitis in SS. Conclusions: A panel of differentially expressed miRNAs were identified in tears of SS model male NOD mice, including some never previously linked to SS. These may have potential utility as diagnostic biomarkers for ocular symptoms of SS; evaluation of the pathways influenced by these dysregulated miRNAs may also provide further insights into SS pathogenesis.

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Evaluation of ionic degradation and slot corrosion of metallic brackets by the action of different dentifrices

Dental Press Journal of Orthodontics ◽

10.1590/s2176-94512013000100019 ◽

2013 ◽

Vol 18 (1) ◽

pp. 86-93

Author(s):

Gustavo Antônio Martins Brandão ◽

Rafael Menezes Simas ◽

Leandro Moreira de Almeida ◽

Juliana Melo da Silva ◽

Marcelo de Castro Meneghim ◽

...

Keyword(s):

Chemical Composition ◽

Artificial Saliva ◽

Positive Control ◽

Negative Control ◽

Sem Analysis ◽

Wilcoxon Test ◽

Surface Polishing ◽

Aluminium Ion ◽

Negative Controls

OBJECTIVE: To evaluate the in vitro ionic degradation and slot base corrosion of metallic brackets subjected to brushing with dentifrices, through analysis of chemical composition by Energy Dispersive Spectroscopy (EDS) and qualitative analysis by Scanning Electron Microscopy (SEM). METHODS: Thirty eight brackets were selected and randomly divided into four experimental groups (n = 7). Two groups (n = 5) worked as positive and negative controls. Simulated orthodontic braces were assembled using 0.019 x 0.025-in stainless steel wires and elastomeric rings. The groups were divided according to surface treatment: G1 (Máxima Proteção Anticáries®); G2 (Total 12®); G3 (Sensitive®); G4 (Branqueador®); Positive control (artificial saliva) and Negative control (no treatment). Twenty eight brushing cycles were performed and evaluations were made before (T0) and after (T1) experiment. RESULTS: The Wilcoxon test showed no difference in ionic concentrations of titanium (Ti), chromium (Cr), iron (Fe) and nickel (Ni) between groups. G2 presented significant reduction (p < 0.05) in the concentration of aluminium ion (Al). Groups G3 and G4 presented significant increase (p < 0.05) in the concentration of aluminium ion. The SEM analysis showed increased characteristics indicative of corrosion on groups G2, G3 and G4. CONCLUSION: The EDS analysis revealed that control groups and G1 did not suffer alterations on the chemical composition. G2 presented degradation in the amount of Al ion. G3 and G4 suffered increase in the concentration of Al. The immersion in artificial saliva and the dentifrice Máxima Proteção Anticáries® did not alter the surface polishing. The dentifrices Total 12®, Sensitive® and Branqueador® altered the surface polishing.

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Interleukin-1 effect on glycemia in the non-obese diabetic mouse at the pre-diabetic stage

Journal of Endocrinology ◽

10.1677/joe.0.1480139 ◽

1996 ◽

Vol 148 (1) ◽

pp. 139-148 ◽

Cited By ~ 25

Author(s):

A Amrani ◽

M Jafarian-Tehrani ◽

P Mormède ◽

S Durant ◽

J-M Pleau ◽

...

Keyword(s):

Insulin Resistance ◽

Type I Diabetes ◽

Interleukin 1 ◽

Nod Mice ◽

Experimental Models ◽

Nod Mouse ◽

Hypoglycemic Effect ◽

Type I ◽

Insulin Effect ◽

Corticosterone Secretion

Abstract Cytokines, particularly interleukin 1 (IL-1) and tumor necrosis factor, are known to induce hypoglycemia in normal rodents or different experimental models of type II diabetes. We investigated, at the pre-diabetic stage, the effect of short-term administration of murine recombinant interleukin-1α (mrIL-1α) on the levels of glucose, insulin and corticosterone in the non-obese diabetic (NOD) mouse, a spontaneous model of type I diabetes. Two-month-old, pre-diabetic NOD mice of both sexes were insensitive to mrIL-1α (12·5 and 50 μg/kg) 2 h after administration, the time at which the maximal decrease (around 50%) was observed in the C57BL/6 mouse strain. Kinetic studies however showed that mrIL-1α lowered glycemia in both sexes of NOD mice, but the effect was limited and delayed. In the NOD and C57BL/6 strains, mrIL-1α had no influence on insulin levels in females, but significantly increased them in males (P<0·0001). Castration of NOD males abrogated the stimulatory effect of mrIL-1α on insulin secretion. Corticosterone secretion was stimulated by mrIL-1α in both sexes of NOD and C57BL/6 mice, and this effect was faster and greater in NOD females than in C57BL/6 females. The incomplete hypoglycemic response to mrIL-1α in females may be attributed to the anti-insulin effect of glucocorticoids, an effect which can be demonstrated when mrIL-1α is administered to adrenalectomized animals or when mrIL-1α is administered together with the glucocorticoid antagonist RU38486. In NOD males, in contrast, glucocorticoids did not play a major role in the limited hypoglycemic response to mrIL-1α, since RU38486 and adrenalectomy were not able to unmask a hypoglycemic effect. Moreover, NOD mice of both sexes were less sensitive than C57BL/6 mice to the hypoglycemic effect of insulin (2·5 U/kg), which suggests some degree of insulin-resistance in NOD mice. With regard to the effect of IL-1 on NOD mouse glycemia, therefore, these results suggest that glucocorticoids and/or androgens, according to the animal's sex, may induce a state of insulin-resistance. Journal of Endocrinology (1996) 148, 139–148

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slowly growing natural populations. Various approaches have been adopted in order to improve the sensitivity. These have included the use of multiple probes labelled with a single fluor (Lee et al. 1993); or labelled with multiple fluors (Trebesius et al. 1994) and enzyme-linked probes or detection systems that allow signal amplification (Lebaron et al. 1997, Schonhuber et al. 1999). The latter indirect approach not only has the potential for signal amplification, but may also be used in natural samples showing high levels of autofluorescence. Any thorough identification method has to include positive and negative controls. False-positive results may either be caused by cells emitting autofluorescence upon excitation or by nonspecific binding of the probe to nontarget cells. Samples should therefore be checked for autofluorescence before hybridization and a negative control with a fluorescent oligonucleotide not complementary to rRNA has to be applied to check for sequence-independent nonspecific binding. Such non-specific binding may be due to interaction of the dye compound of the probe with hydrophobic cell components. Failures to detect cells containing target sequences (false-negatives) may originate from cells with either low cellular ribosome content or limited permeability of the cell periphery for the fluorescent probe (Manz et al. 1992). With the rapidly expanding database of 16S rRNA sequences, the problem of probe specificity has become more apparent and the design of probes is becoming increasingly difficult. These problems are also applicable to PCR and other oligonucleotide-dependent techniques. The problem of probe specificity may be overcome by using multiple specific oligonucleotide probes targeting different sites on the rRNA molecule and labelled with different fluorochromes. While a single oligonucleotide target sequence may be found in a number of related taxa, the probability that target sites for three designed oligonucleotides are found in a nontarget organism is, however, much reduced.

Recent Advances in Marine Biotechnology, Vol. 8 ◽

10.1201/9781482279986-25 ◽

2003 ◽

pp. 232-232

Keyword(s):

Signal Amplification ◽

Specific Binding ◽

Natural Populations ◽

Negative Control ◽

Target Sequence ◽

Cell Periphery ◽

Nonspecific Binding ◽

Detection Systems ◽

Cell Components ◽

Negative Controls

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Effect of Sodium Carboxymethylcellulose and Sorbitol on Anti-Peptic Ulcer Activity of Anredera cordifolia Leaves Extract

Pharmacology and Clinical Pharmacy Research ◽

10.15416/pcpr.v4i1.21392 ◽

2019 ◽

Vol 4 (1) ◽

pp. 16

Author(s):

Maria Ulfah ◽

Revika Rachmaniar ◽

Egi MR. Sudrajat ◽

Rida W. Fadla ◽

Hary S. Pinuji

Keyword(s):

Body Weight ◽

Peptic Ulcer ◽

Normal Control ◽

Wistar Rat ◽

Positive Control ◽

Negative Control ◽

Sodium Carboxymethylcellulose ◽

Freeze Dried ◽

Male Wistar Rats ◽

Negative Controls

Anredera cordifolia or binahong is one of the Indonesian medicinal plants that is used to treat peptic ulcer. The purpose of this study was to evaluate the effect of the addition of sodium carboxymethylcellulose (CMC) and sorbitol on anti-peptic ulcer activity of A. cordifolia leaves extracts in male Wistar rats. The plants were extracted using decoction method and freeze dried. Three liquid formulas were used i.e., i) a combination of sodium CMC and sorbitol; ii) only sorbitol; iii) extract only. The rats were divided into 6 groups, i.e., positive control (sucralfate 35 mg/kg body weight); negative control (80% ethanol); normal control; and 3 formulas. After the administration of the liquid formula, all groups, except normal control, were given 80% ethanol (l5 ml/kg body weight) to induce peptic ulcer. Antipeptic ulcer activity was evaluated using direct observation on rats gastric mucosa, and histopathology assessment. The result showed that the strongest anti-peptic ulcer was shown by sorbitol only (96.95% inhibition), followed by the combination of sodium CMC and sorbitol (92.68% inhibition). The formula which only contained extract showed only 31.70% inhibition. Statistical analysis showed significant differences between formula 1 and 2 with negative controls. In conclusion, A. cordifolia leaves extract with the addition of sorbitol showed the strongest anti-peptic ulcer activity. Keyword: Anredera cordifolia, peptic ulcer, suspense, Wistar rat.

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Gene Transfection and Expression of Transforming Growth Factor-β1 in Nonobese Diabetic Mouse Islets Protects β-Cells in Syngeneic Islet Grafts from Autoimmune Destruction

Cell Transplantation ◽

10.3727/000000002783985503 ◽

2002 ◽

Vol 11 (6) ◽

pp. 519-528 ◽

Cited By ~ 24

Author(s):

Wilma L. Suarez-Pinzon ◽

Yvonne Marcoux ◽

Aziz Ghahary ◽

Alex Rabinovitch

Keyword(s):

Transforming Growth Factor ◽

Islet Cells ◽

Gene Transfection ◽

Nod Mice ◽

Nod Mouse ◽

Β Cells ◽

Nonobese Diabetic ◽

Autoimmune Destruction ◽

Mouse Islets ◽

Tgf Β1

Nonobese diabetic (NOD) mice develop diabetes and destroy syngeneic islet grafts through an autoimmune response. Because transforming growth factor (TGF)-β1 downregulates immune responses, we tested whether overexpression of TGF-β1 by gene transfection of NOD mouse islets could protect β-cells in islet grafts from autoimmune destruction. NOD mouse islet cells were transfected with an adenoviral DNA expression vector encoding porcine latent TGF-β1 (Ad TGF- β1) or the adenoviral vector alone (control Ad vector). The frequency of total islet cells expressing TGF-1 protein was increased from 12±1% in control Ad vector-transfected cells to 89 ± 4% in Ad TGF-β1-transfected islet cells, and the frequency of β-cells that expressed TGF-β1 was increased from 12 ± 1% to 60 ± 7%. Also, secretion of TGF-β1 was significantly increased in islets that overexpressed TGF-β1. Ad TGF-β1-transfected NOD mouse islets that overexpressed TGF-β1 prevented diabetes recurrence after transplantation into diabetic NOD mice for a median of 22 days compared with only 7 days for control Ad vector-transfected islets (p = 0.001). Immunohistochemical examination of the islet grafts revealed significantly more TGF-β1+ cells and insulin+ cells and significantly fewer CD45+ leukocytes in Ad TGF-β1-transfected islet grafts. Also, islet β-cell apoptosis was significantly decreased whereas apoptosis of graft-infiltrating leukocytes was significantly increased in Ad TGF-β1-transfected islet grafts. These observations demonstrate that overexpression of TGF-β1, by gene transfection of NOD mouse islets, protects islet β-cells from apoptosis and autoimmune destruction and delays diabetes recurrence after islet transplantation.

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Abstract 2819: Cross-correlation Analysis of Myocardial Velocity is Superior to Conventional Time-to-Peak Parameters for Quantifying Left Ventricular Dyssynchrony

Circulation ◽

10.1161/circ.116.suppl_16.ii_627 ◽

2007 ◽

Vol 116 (suppl_16) ◽

Author(s):

Brandon K Fornwalt ◽

Takeshi Arita ◽

Mohit Bhasin ◽

George Voulgaris ◽

John D Merlino ◽

...

Keyword(s):

Cross Correlation ◽

Positive Control ◽

Negative Control ◽

Left Ventricular ◽

Left Ventricular Dyssynchrony ◽

Control Group ◽

Time To Peak ◽

Data Points ◽

Negative Controls ◽

Threshold Line

Background- A recent study showed that the most commonly used Tissue Doppler imaging (TDI) parameters to diagnose left ventricular dyssynchrony agree only 50% of the time. Most of these parameters require calculation of the ``time-to-peak” myocardial velocity. This ``time-to-peak” based analysis utilizes only one of >100 data points collected per heart cycle. Methods- We developed and tested a new dyssynchrony parameter, cross-correlation delay (XCD), that utilizes all velocity data points from 3 consecutive beats (~420 points). We hypothesized that XCD would be superior to existing methods at diagnosing dyssynchrony. We tested XCD on 11 members of a positive control group (echocardiographic responders to cardiac resynchronization therapy) and 12 members of a negative control group (normal echocardiogram and 12-lead ECG). We compared XCD to septal-to-lateral delay in time-to-peak (SLD), maximum difference in the basal 2- or 4-chamber times-to-peak (MaxDiff) and standard deviation of the 12 basal and mid-wall times-to-peak (Ts-SD). Results- An XCD threshold of 31ms discriminated between positive and negative controls with 100% sensitivity and specificity (Figure 1 ). SLD, MaxDiff and Ts-SD showed sensitivities of 36, 55 and 100% and specificities of 50, 42 and 50%, respectively. ROC analysis showed XCD and Ts-SD were superior to SLD and MaxDiff in discriminating between positive and negative controls (p<0.01). XCD was the only parameter which decreased after resynchronization in the positive controls (from 160±88ms to 69±61ms, p=0.003). Conclusion- XCD is superior to existing parameters at discriminating patients with LV dyssynchrony from those with normal function. Figure 1. XCD shows the greatest discrimination between positive and negative controls. Dyssynchrony values for each positive control are shown as x’s and values for each negative control are shown as circels. Different dyssynchrony parameters are shown in each subplot (A-D). Threshold values to diagnose dyssynchrony are plotted as horizontal lines in each figure. Note that x’s above the threshold line represent false positives while circles below the threshold line represent false negatives.

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Tolerogenic Delivery of a Hybrid Insulin Peptide Markedly Prolongs Islet Graft Survival in the Non-Obese Diabetic (NOD) Mouse

10.2337/figshare.17188097 ◽

2022 ◽

Author(s):

Braxton L. Jamison ◽

James E. DiLisio ◽

K. Scott Beard ◽

Tobias Neef ◽

Brenda Bradley ◽

...

Keyword(s):

T Cells ◽

Graft Survival ◽

Tolerance Induction ◽

Nod Mice ◽

Disease Recurrence ◽

Nod Mouse ◽

Islet Graft ◽

Specific Effector ◽

Syngeneic Islet Transplantation ◽

Specific Tolerance

The induction of antigen (Ag)-specific tolerance and replacement of islet β-cells are major ongoing goals for the treatment of Type 1 Diabetes (T1D). Our group previously showed that a hybrid insulin peptide (2.5HIP) is a critical autoantigen for diabetogenic CD4+ T cells in the non-obese diabetic (NOD) mouse model. In this study, we investigated whether induction of Ag-specific tolerance using 2.5HIP-coupled tolerogenic nanoparticles (NPs) could protect diabetic NOD mice from disease recurrence upon syngeneic islet transplantation. Islet graft survival was significantly prolonged in mice treated with 2.5HIP NPs, but not NPs containing the insulin B chain peptide 9-23. Protection in 2.5HIP NP-treated mice was attributed both to the simultaneous induction of anergy in 2.5HIP-specific effector T cells and to the expansion of Foxp3+ regulatory T cells specific for the same antigen. Notably, our results indicate that effector function of graft-infiltrating CD4+ and CD8+ T cells specific for other β-cell epitopes was significantly impaired, suggesting a novel mechanism of therapeutically induced linked suppression. This work establishes that tolerance induction with a hybrid insulin peptide can delay recurrent autoimmunity in NOD mice, which could inform the development of an Ag-specific therapy for T1D.

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Essential oils as tick repellents on clothing

Experimental and Applied Acarology ◽

10.1007/s10493-019-00422-z ◽

2019 ◽

Vol 79 (2) ◽

pp. 209-219 ◽

Cited By ~ 3

Author(s):

Oliver Soutar ◽

Freya Cohen ◽

Richard Wall

Keyword(s):

Essential Oils ◽

No Reduction ◽

Positive Control ◽

Negative Control ◽

Tick Abundance ◽

Spearmint Oil ◽

Oregano Oil ◽

Negative Controls ◽

Tick Repellents

Abstract Essential oils show promise as natural alternatives to synthetic tick repellents, but few studies have investigated their repellent efficacy in vivo or under field conditions. Here, blanket-drags and standardised walks were employed to evaluate tick acquisition by 1 m2 cotton blankets or cotton trousers, respectively, in woodland edge habitats of known high tick abundance. Blankets and trousers had been treated with one of 5% oregano, rosemary, spearmint or thyme oils, 20% DEET (N,N-diethyl-3-methylbenzamide) (positive control) or ethanol excipient-only (negative control). The number of ticks present on the blankets or trousers differed significantly between treatments: spearmint oil treatments resulted in significantly fewer ticks than the negative controls for both blankets and trousers and significantly fewer ticks were present on the oregano oil treated blankets. For ticks that did attach to the trousers, the rate of drop off within 3 min was significantly higher for trousers treated with spearmint oil or thyme oil than ethanol, oregano oil and rosemary oil. No reduction in repellence was detected over a 24 h period between treatment and testing. The results suggest that 5% oregano and spearmint oils exhibit potential as natural clothing repellents, with an effective equivalence to 20% DEET.

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Antimicrobial Activity of an Amnion-Chorion Membrane to Oral Microbes

International Journal of Dentistry ◽

10.1155/2019/1269534 ◽

2019 ◽

Vol 2019 ◽

pp. 1-7 ◽

Cited By ~ 1

Author(s):

Haroon Ashraf ◽

Kerri Font ◽

Charles Powell ◽

Michael Schurr

Keyword(s):

Bacterial Species ◽

Healing Process ◽

Antimicrobial Properties ◽

Positive Control ◽

Negative Control ◽

Collagen Membrane ◽

Wound Healing Process ◽

Bacterial Strains ◽

Oral Microbes ◽

Negative Controls

Objective. The aim of this study was to evaluate wound biomodification by assessing antimicrobial properties present within a human-derived composite amnion-chorion membrane (ACM). Methods. Membranes analyzed were the human-derived ACM BioXclude™ and the porcine-derived collagen membrane Bio-Gide®. Paper discs with and without tetracycline served as positive and negative controls, respectively. The same number of colony-forming units per milliliter for each bacterial species (Aggregatibacter actinomycetemcomitans, Streptococcus mutans, and Streptococcus oralis) was inoculated on each of the discs. Discs from each group were removed at 12 and 24 hours and sonicated to remove the bacteria off the membranes. A serial dilution was performed to quantify bacterial growth. Results. The ACM inhibited growth at all time points, with all bacterial strains, identical to the negative control tetracycline discs. The collagen membrane and positive controls did not inhibit growth of any of the bacterial species throughout the 24-hour study period. P<0.05 for microbial growth on ACM or negative control vs. either collagen membrane or positive control. Conclusion. ACM was proven to be as bactericidal as paper discs inoculated with tetracycline at its minimum bactericidal concentration. The ACM bactericidal property may be beneficial in the early wound healing process.

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KULIT BUAH JERUK LIMAU (Citrus amblycarpa (Hassk.) Osche) SEBAGAI ANALGESIK

Jurnal Kimia ◽

10.24843/jchem.2020.v14.i01.p05 ◽

2020 ◽

pp. 24

Author(s):

R. A. I. K. Maharani ◽

N. K. Cahyaningsih ◽

M. D. Abimanyu ◽

K. W. Astuti

Keyword(s):

Body Weight ◽

Analgesic Activity ◽

Treatment Options ◽

Ethanol Extract ◽

Positive Control ◽

Negative Control ◽

Fruit Peel ◽

Hot Plate ◽

Inflammatory Drugs ◽

Negative Controls

Nonsteroidal anti-inflammatory drugs (NSAIDs) are the treatment options for relieving pain. However, long-term use can trigger gastrointestinal bleeding. Therefore, alternative analgesics which have the same therapeutic effect with lower side effects are needed. Limau (Citrus amblycarpa) is an empirical drug for tingling and cramping. The aim of the study is to determine the analgesic activity of ethanol extract of C. amblycarpa fruit peel. The method used in testing analgesic activity is the Hot Plate method. The study was conducted by dividing 30 mice into 6 groups. The group given CMC-Na 1% was used as a negative control, the group given suspension of sodium diclofenac dose of 6.5 mg/kg of body weight was used as a positive control, and the group given suspension of ethanol extract of C. amblycarpa fruit peel with dose variations 100, 300 and 600 mg/kg of body weight. The test animals were placed on top of the Hot Plate with a temperature of 70°C at 30 minutes after giving suspension test and the response time of mice to heat was observed every 30 minutes for 3 hours with cut off time 15 second. Based on the test results, it can be concluded that the administration of ethanol extract of C. amblycarpa fruit peel with 100, 300 and 600 mg/kg of body weight gave analgesic activity on mice compared to the negative controls (CMC-Na 1%). Keywords: C. amblycarpa, Fruit Peel, Analgesics, Hot Plate

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