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Article ; Online: Single-cell transcriptomic analysis of the identity and function of fibro/adipogenic progenitors in healthy and dystrophic muscle.

Uapinyoying, Prech / Hogarth, Marshall / Battacharya, Surajit / Mázala, Davi A G / Panchapakesan, Karuna / Bönnemann, Carsten G / Jaiswal, Jyoti K

iScience

2023 Volume 26, Issue 8, Page(s) 107479

Abstract: Fibro/adipogenic progenitors (FAPs) are skeletal muscle stromal cells that support regeneration of injured myofibers and their maintenance in healthy muscles. FAPs are related to mesenchymal stem cells (MSCs/MeSCs) found in other adult tissues, but there ...

Abstract	Fibro/adipogenic progenitors (FAPs) are skeletal muscle stromal cells that support regeneration of injured myofibers and their maintenance in healthy muscles. FAPs are related to mesenchymal stem cells (MSCs/MeSCs) found in other adult tissues, but there is poor understanding of the extent of similarity between these cells. Using single-cell RNA sequencing (scRNA-seq) datasets from multiple mouse tissues, we have performed comparative transcriptomic analysis. This identified remarkable transcriptional similarity between FAPs and MeSCs, confirmed the suitability of PDGFRα as a reporter for FAPs, and identified extracellular proteolysis as a new FAP function. Using PDGFRα as a cell surface marker, we isolated FAPs from healthy and dysferlinopathic mouse muscles and performed scRNA-seq analysis. This revealed decreased FAP-mediated Wnt signaling as a potential driver of FAP dysfunction in dysferlinopathic muscles. Analysis of FAPs in dysferlin- and dystrophin-deficient muscles identified a relationship between the nature of muscle pathology and alteration in FAP gene expression.
Language	English
Publishing date	2023-07-23
Publishing country	United States
Document type	Journal Article
ISSN	2589-0042
ISSN (online)	2589-0042
DOI	10.1016/j.isci.2023.107479
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Article ; Online: A cross-sectional analysis of the urine microbiome of children with neuropathic bladders.

Forster, Catherine S / Panchapakesan, Karuna / Stroud, Crystal / Banerjee, Payal / Gordish-Dressman, Heather / Hsieh, Michael H

Journal of pediatric urology

2020 Volume 16, Issue 5, Page(s) 593.e1–593.e8

Abstract: Background: Distinguishing a urinary tract infection (UTI) from asymptomatic bacteriuria (ASB) in children with neuropathic bladders is difficult. Currently used markers of infection, such as the routine urinalysis, lack specificity for UTI in this ... ...

Abstract	Background: Distinguishing a urinary tract infection (UTI) from asymptomatic bacteriuria (ASB) in children with neuropathic bladders is difficult. Currently used markers of infection, such as the routine urinalysis, lack specificity for UTI in this population. The urinary microbiome may help differentiate these states. Objective: The objective of this work was to describe the baseline microbiome in children with neuropathic bladders, and to determine if differences exist among the urine microbiomes of children with neuropathic bladders who have negative urine cultures, ASB, or UTI. Study design: This is a cross-sectional study of children with neuropathic bladders who use clean intermittent catheterization for bladder management who had a urine culture sent as part of clinical management. Residual urine, initially collected via catheter for urine culture, was obtained for use in this work. Microbial DNA was isolated, and the V4 region of the 16SrRNA gene sequenced. The relative abundance of each bacteria was measured in each group. Alpha diversity, measured by Chao1 and the Shannon Diversity Index, was also measured in each group. PERMANOVA was used to compare the microbiota between groups. Results: 36 children with neuropathic bladders were included in this study (UTI = 11, ASB = 19, negative cultures = 4). The most abundant bacteria were unspecified Enterobacteriaceae, Klebsiella, Staphylococcus, Streptococcus, and Enterococcus. Children who catheterize their urethra have a higher proportion of Staphylococcus, while the urine microbiome of those who catheterize through a Mitrofanoff consists predominantly of members of the family Enterobacteriaceae. Given the low numbers of patients with Mitrofanoffs and augmented bladders, we did not statistically compare the urine microbiomes between these patients. There was no difference in either alpha diversity or the overall microbiota between children with neuropathic bladders with UTI, ASB, and negative cultures. Discussion: In this pilot cohort of children with neuropathic bladders, bacteria that are members of the family Enterobacteriaceae are the most predominant bacteria in the urine microbiomes. There was no difference in the urine microbiome between those with UTI, ASB, and negative cultures. Route of catheterization may affect the composition of the urine microbiome, although due to limited sample size, this was not confirmed statistically. Conclusion: There was no difference in the urine microbiome between patients with negative urine cultures, ASB, and UTI. Further work is needed to determine if the urine microbiome varies based on either the route of catheterization or the presence of augmented bladder.
MeSH term(s)	Bacteriuria ; Child ; Cross-Sectional Studies ; Humans ; Male ; Microbiota ; Urinary Bladder, Neurogenic/therapy ; Urinary Tract Infections ; Urine
Language	English
Publishing date	2020-02-19
Publishing country	England
Document type	Journal Article
ZDB-ID	2237683-5
ISSN	1873-4898 ; 1477-5131
ISSN (online)	1873-4898
ISSN	1477-5131
DOI	10.1016/j.jpurol.2020.02.005
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Zs.A 6387: Show issues

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Article ; Online: A long-read RNA-seq approach to identify novel transcripts of very large genes.

Uapinyoying, Prech / Goecks, Jeremy / Knoblach, Susan M / Panchapakesan, Karuna / Bonnemann, Carsten G / Partridge, Terence A / Jaiswal, Jyoti K / Hoffman, Eric P

Genome research

2020 Volume 30, Issue 6, Page(s) 885–897

Abstract: RNA-seq is widely used for studying gene expression, but commonly used sequencing platforms produce short reads that only span up to two exon junctions per read. This makes it difficult to accurately determine the composition and phasing of exons within ... ...

Abstract	RNA-seq is widely used for studying gene expression, but commonly used sequencing platforms produce short reads that only span up to two exon junctions per read. This makes it difficult to accurately determine the composition and phasing of exons within transcripts. Although long-read sequencing improves this issue, it is not amenable to precise quantitation, which limits its utility for differential expression studies. We used long-read isoform sequencing combined with a novel analysis approach to compare alternative splicing of large, repetitive structural genes in muscles. Analysis of muscle structural genes that produce medium (
MeSH term(s)	Alternative Splicing ; Computational Biology/methods ; Exons ; Gene Expression Profiling/methods ; High-Throughput Nucleotide Sequencing ; Humans ; Molecular Sequence Annotation ; Organ Specificity/genetics ; RNA, Messenger ; Repetitive Sequences, Nucleic Acid ; Sequence Analysis, RNA ; Transcriptome
Chemical Substances	RNA, Messenger
Language	English
Publishing date	2020-07-06
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	1284872-4
ISSN	1549-5469 ; 1088-9051 ; 1054-9803
ISSN (online)	1549-5469
ISSN	1088-9051 ; 1054-9803
DOI	10.1101/gr.259903.119
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Zs.A 3729: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (2.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MG 8: Show issues

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Article ; Online: Vamorolone, a dissociative steroidal compound, reduces collagen antibody-induced joint damage and inflammation when administered after disease onset.

Damsker, Jesse M / Cornish, Michaelyn R / Kanneboyina, Priya / Kanneboyina, Ila / Yu, Qing / Lipson, Rachel / Phadke, Aditi / Knoblach, Susan M / Panchapakesan, Karuna / Morales, Melissa / Fiorillo, Alyson A / Partridge, Terence / Nagaraju, Kanneboyina

Inflammation research : official journal of the European Histamine Research Society ... [et al.

2019 Volume 68, Issue 11, Page(s) 969–980

Abstract: Objective and design: The objective of this study was to assess the effect of vamorolone, a first-in-class dissociative steroidal compound, to inhibit inflammation when administered after disease onset in the murine collagen antibody-induced arthritis ... ...

Abstract	Objective and design: The objective of this study was to assess the effect of vamorolone, a first-in-class dissociative steroidal compound, to inhibit inflammation when administered after disease onset in the murine collagen antibody-induced arthritis model of arthritis. Animals: 84 DBA1/J mice were used in this study (n = 12 per treatment group). Treatment: Vamorolone or prednisolone was administered orally after disease onset for a duration of 7 days. Methods: Disease score and bone erosion were assessed using previously described scoring systems. Cytokines were measured in joints via immunoassay, and joint cathepsin B activity (marker of inflammation) was assessed using optical imaging of joints on live mice. Results: We found that vamorolone treatment led to a reduction of several disease parameters including disease score, joint inflammation, and the presence of pro-inflammatory mediators to a degree similar of that observed with prednisolone treatment. More importantly, histopathological analysis of affected joints showed that vamorolone treatment significantly reduced the degree of bone erosion while this bone-sparing property was not observed with prednisolone treatment at any of the tested doses. Conclusions: While many intervention regimens in other studies are administered prior to disease onset in animal models, the current study involves delivery of the potential therapeutic after disease onset. Based on the findings, vamorolone may offer an efficacious, yet safer alternative to conventional steroidal compounds in the treatment of rheumatoid arthritis and other inflammatory diseases.
MeSH term(s)	Animals ; Anti-Inflammatory Agents/therapeutic use ; Antibodies, Monoclonal/immunology ; Arthritis, Experimental/drug therapy ; Arthritis, Experimental/immunology ; Arthritis, Experimental/pathology ; Collagen Type II/immunology ; Cytokines/immunology ; Joints/drug effects ; Joints/immunology ; Joints/pathology ; Lipopolysaccharides ; Male ; Mice, Inbred DBA ; Pregnadienediols/therapeutic use
Chemical Substances	Anti-Inflammatory Agents ; Antibodies, Monoclonal ; Collagen Type II ; Cytokines ; Lipopolysaccharides ; Pregnadienediols ; VBP15 compound
Language	English
Publishing date	2019-08-24
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	1221794-3
ISSN	1420-908X ; 1023-3830
ISSN (online)	1420-908X
ISSN	1023-3830
DOI	10.1007/s00011-019-01279-z
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Zs.A 675: Show issues

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Article ; Online: VBP15, a novel dissociative steroid compound, reduces NFκB-induced expression of inflammatory cytokines in vitro and symptoms of murine trinitrobenzene sulfonic acid-induced colitis.

Damsker, Jesse M / Conklin, Laurie S / Sadri, Soheil / Dillingham, Blythe C / Panchapakesan, Karuna / Heier, Christopher R / McCall, John M / Sandler, Anthony D

Inflammation research : official journal of the European Histamine Research Society ... [et al.

2016 Volume 65, Issue 9, Page(s) 737–743

Abstract: Objective and design: The goal of this study was to assess the capacity of VBP15, a dissociative steroidal compound, to reduce pro-inflammatory cytokine expression in vitro, to reduce symptoms of colitis in the trinitrobenzene sulfonic acid-induced ... ...

Abstract	Objective and design: The goal of this study was to assess the capacity of VBP15, a dissociative steroidal compound, to reduce pro-inflammatory cytokine expression in vitro, to reduce symptoms of colitis in the trinitrobenzene sulfonic acid-induced murine model, and to assess the effect of VBP15 on growth stunting in juvenile mice. Materials: In vitro studies were performed in primary human intestinal epithelial cells. Colitis was induced in mice by administering trinitrobenzene sulfonic acid. Growth stunting studies were performed in wild type outbred mice. Treatment: Cells were treated with VBP15 or prednisolone (10 μM) for 24 h. Mice were subjected to 3 days of VBP15 (30 mg/kg) or prednisolone (30 mg/kg) in the colitis study. In the growth stunting study, mice were subjected to VBP15 (10, 30, 45 mg/kg) or prednisolone (10 mg/kg) for 5 weeks. Methods: Cytokines were measured by PCR and via Luminex. Colitis symptoms were evaluated by assessing weight loss, intestinal blood, and stool consistency. Growth stunting was assessed using an electronic caliper. Results: VBP15 significantly reduced the in vitro production of CCL5 (p < 0.001) IL-6 (p < 0.001), IL-8 (p < 0.05) and reduced colitis symptoms (p < 0.05). VBP15 caused less growth stunting than prednisolone (p < 0.001) in juvenile mice. Conclusion: VBP15 may reduce symptoms of IBD, while decreasing or avoiding detrimental side effects.
MeSH term(s)	Animals ; Anti-Inflammatory Agents/pharmacology ; Anti-Inflammatory Agents/therapeutic use ; Body Size/drug effects ; Cells, Cultured ; Colitis/chemically induced ; Colitis/drug therapy ; Colitis/metabolism ; Cytokines/genetics ; Cytokines/metabolism ; Disease Models, Animal ; Epithelial Cells/drug effects ; Epithelial Cells/metabolism ; Female ; Humans ; Male ; Mice, Inbred BALB C ; NF-kappa B/metabolism ; Pregnadienediols/pharmacology ; Pregnadienediols/therapeutic use ; Trinitrobenzenesulfonic Acid
Chemical Substances	Anti-Inflammatory Agents ; Cytokines ; NF-kappa B ; Pregnadienediols ; VBP15 compound ; Trinitrobenzenesulfonic Acid (8T3HQG2ZC4)
Language	English
Publishing date	2016-09
Publishing country	Switzerland
Document type	Journal Article
ZDB-ID	1221794-3
ISSN	1420-908X ; 1023-3830
ISSN (online)	1420-908X
ISSN	1023-3830
DOI	10.1007/s00011-016-0956-8
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Zs.A 675: Show issues

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Article ; Online: PAC1R Genotype to Phenotype Correlations in Autism Spectrum Disorder.

Goodrich, Meredith / Armour, Anna Chelsea / Panchapakesan, Karuna / You, Xiaozhen / Devaney, Joseph / Knoblach, Susan / Sullivan, Catherine A W / Herrero, Maria Jesus / Gupta, Abha R / Vaidya, Chandan J / Kenworthy, Lauren / Corbin, Joshua G

Autism research : official journal of the International Society for Autism Research

2018 Volume 12, Issue 2, Page(s) 200–211

Abstract: Amygdala dysfunction has been implicated in numerous neurodevelopmental disorders, including autism spectrum disorder (ASD). Previous studies in mice and humans, respectively, have linked Pac1r/PAC1R function to social behavior and PTSD-susceptibility. ... ...

Abstract	Amygdala dysfunction has been implicated in numerous neurodevelopmental disorders, including autism spectrum disorder (ASD). Previous studies in mice and humans, respectively, have linked Pac1r/PAC1R function to social behavior and PTSD-susceptibility. Based on this connection to social and emotional processing and the central role played by the amygdala in ASD, we examined a putative role for PAC1R in social deficits in ASD and determined the pattern of gene expression in the developing mouse and human amygdala. We reveal that Pac1r/PAC1R is expressed in both mouse and human amygdala from mid-neurogenesis through early postnatal stages, critical time points when altered brain trajectories are hypothesized to unfold in ASD. We further find that parents of autistic children carrying a previously identified PTSD-risk genotype (CC) report greater reciprocal social deficits compared to those carrying the non-risk GC genotype. Additionally, by exploring resting-state functional connectivity differences in a subsample of the larger behavioral sample, we find higher functional connectivity between the amygdala and right middle temporal gyrus in individuals with the CC risk genotype. Thus, using multimodal approaches, our data reveal that the amygdala-expressed PAC1R gene may be linked to severity of ASD social phenotype and possible alterations in brain connectivity, therefore potentially acting as a modifier of amygdala-related phenotypes. Autism Res 2019, 12: 200-211 © 2018 International Society for Autism Research, Wiley Periodicals, Inc. LAY SUMMARY: In this multimodal study across mouse and human, we examined expression patterns of Pac1r/PAC1R, a gene implicated in social behavior, and further explored whether a previously identified human PTSD-linked mutation in PAC1R can predict brain connectivity and social deficits in ASD. We find that PAC1R is highly expressed in the both the mouse and human amygdala. Furthermore, our human data suggest that PAC1R genotype is linked to severity of social deficits and functional amygdala connectivity in ASD.
MeSH term(s)	Adolescent ; Animals ; Autism Spectrum Disorder/genetics ; Autism Spectrum Disorder/physiopathology ; Brain/diagnostic imaging ; Brain/physiopathology ; Brain Mapping/methods ; Child ; Disease Models, Animal ; Female ; Genotype ; Humans ; Magnetic Resonance Imaging/methods ; Male ; Mice ; Mice, Inbred C57BL ; Phenotype ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I/genetics
Chemical Substances	ADCYAP1R1 protein, human ; Receptors, Pituitary Adenylate Cyclase-Activating Polypeptide, Type I
Language	English
Publishing date	2018-12-17
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, Non-U.S. Gov't
ZDB-ID	2481338-2
ISSN	1939-3806 ; 1939-3792
ISSN (online)	1939-3806
ISSN	1939-3792
DOI	10.1002/aur.2051
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Zs.A 6785: Show issues

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Article ; Online: Association of brain injury and neonatal cytokine response during therapeutic hypothermia in newborns with hypoxic-ischemic encephalopathy.

Orrock, Janet E / Panchapakesan, Karuna / Vezina, Gilbert / Chang, Taeun / Harris, Kari / Wang, Yunfei / Knoblach, Susan / Massaro, An N

Pediatric research

2015 Volume 79, Issue 5, Page(s) 742–747

Abstract: Background: Cytokines have been proposed as mediators of neonatal brain injury via neuroinflammatory pathways triggered by hypoxia-ischemia. Limited data are available on cytokine profiles in larger cohorts of newborns with hypoxic-ischemic ... ...

Abstract	Background: Cytokines have been proposed as mediators of neonatal brain injury via neuroinflammatory pathways triggered by hypoxia-ischemia. Limited data are available on cytokine profiles in larger cohorts of newborns with hypoxic-ischemic encephalopathy (HIE) undergoing therapeutic hypothermia (TH). Methods: Serum cytokines interleukin (IL)-1β, IL-2, IL-4, IL-6, IL-8, IL-10, IL-13, tumor necrosis factor-α, and interferon-γ were measured in newborns with HIE at 24 and 72 h of TH. Differences between infants with favorable (survivors with mild/no magnetic resonance imaging (MRI) injury) vs. adverse outcome (death or moderate/severe MRI injury) were compared using mixed models to adjust for covariates. Results: Data from 36 term newborns with HIE (favorable outcome: n = 20, adverse outcome: n = 16) were evaluated. Cytokines IL-1β, IL-2, IL-6, IL-8, IL-10, and IL-13 were elevated in the adverse relative to favorable outcome group at 24 h. IL-6 remained significantly elevated in the adverse outcome group at 72 h. IL-6 and IL-10 remained significantly associated with outcome group after controlling for covariates. Conclusion: Inflammatory cytokines are elevated in HIE newborns with brain injury by MRI. In particular, IL-6 and IL-10 were associated with adverse outcomes after controlling for baseline characteristics and severity of presentation. These data suggest that cytokine response may identify infants in need of additional neuroprotective interventions.
MeSH term(s)	Brain Injuries/blood ; Cohort Studies ; Cytokines/blood ; Cytokines/metabolism ; Female ; Gestational Age ; Humans ; Hypothermia, Induced ; Hypoxia-Ischemia, Brain/blood ; Hypoxia-Ischemia, Brain/therapy ; Infant, Newborn ; Inflammation ; Magnetic Resonance Imaging ; Male ; Time Factors
Chemical Substances	Cytokines
Language	English
Publishing date	2015-12-30
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	4411-8
ISSN	1530-0447 ; 0031-3998
ISSN (online)	1530-0447
ISSN	0031-3998
DOI	10.1038/pr.2015.280
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Zs.A 564: Show issues			Location: Je nach Verfügbarkeit (siehe Angabe bei Bestand) bis Jg. 1994: Bestellungen von Artikeln über das Online-Bestellformular Jg. 1995 - 2021: Lesesall (1.OG) ab Jg. 2022: Lesesaal (EG)
Zs.MO 373: Show issues

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Article ; Online: CK-MM Polymorphism is Associated With Physical Fitness Test Scores in Military Recruits.

Sprouse, Courtney / Tosi, Laura L / Gordish-Dressman, Heather / Abdel-Ghani, Mai S / Panchapakesan, Karuna / Niederberger, Brenda / Devaney, Joseph M / Kelly, Karen R

Military medicine

2015 Volume 180, Issue 9, Page(s) 1001–1005

Abstract: Objective: Muscle-specific creatine kinase is thought to play an integral role in maintaining energy homeostasis by providing a supply of creatine phosphate. The genetic variant, rs8111989, contributes to individual differences in physical performance, ... ...

Abstract	Objective: Muscle-specific creatine kinase is thought to play an integral role in maintaining energy homeostasis by providing a supply of creatine phosphate. The genetic variant, rs8111989, contributes to individual differences in physical performance, and thus the purpose of this study was to determine if rs8111989 variant is predictive of Physical Fitness Test (PFT) scores in male, military infantry recruits. Methods: DNA was extracted from whole blood, and genotyping was performed in 176 Marines. Relationships between PFT measures (run, sit-ups, and pull-ups) and genotype were determined. Results: Participants with 2 copies of the T allele for rs8111989 variant had higher PFT scores for run time, pull-ups, and total PFT score. Specifically, participants with 2 copies of the TT allele (variant) (n = 97) demonstrated an overall higher total PFT score as compared with those with one copy of the C allele (n = 79) (TT: 250 ± 31 vs. Cc/ct: 238 ± 31; p = 0.02), run score (TT: 82 ± 10 vs. Cc/ct: 78 ± 11; p = 0.04) and pull-up score (TT: 78 ± 11 vs. Cc/ct: 65 ± 21; p = 0.04) or those with the CC/CT genotype. Conclusion: These results demonstrate an association between physical performance measures and genetic variation in the muscle-specific creatine kinase gene (rs8111989).
MeSH term(s)	Adolescent ; Creatine Kinase, MM Form/genetics ; Exercise Test ; Genotype ; Humans ; Male ; Military Personnel ; Physical Fitness ; Polymorphism, Single Nucleotide ; United States ; Young Adult
Chemical Substances	Creatine Kinase, MM Form (EC 2.7.3.2)
Language	English
Publishing date	2015-09
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural ; Research Support, U.S. Gov't, Non-P.H.S.
ZDB-ID	391061-1
ISSN	1930-613X ; 0026-4075
ISSN (online)	1930-613X
ISSN	0026-4075
DOI	10.7205/MILMED-D-14-00331
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Article ; Online: Relationship of the Middle Ear Effusion Microbiome to Secretory Mucin Production in Pediatric Patients With Chronic Otitis Media.

Krueger, Anna / Val, Stéphanie / Pérez-Losada, Marcos / Panchapakesan, Karuna / Devaney, Joe / Duah, Vanessa / DeMason, Christine / Poley, Marian / Rose, Mary / Preciado, Diego

The Pediatric infectious disease journal

2016 Volume 36, Issue 7, Page(s) 635–640

Abstract: Background: Acute otitis media, an infection of the middle ear, can become chronic after multiple episodes. Microbial influence on chronic otitis media remains unclear. It has been reported that mucin glycoproteins are required for middle ear immune ... ...

Abstract	Background: Acute otitis media, an infection of the middle ear, can become chronic after multiple episodes. Microbial influence on chronic otitis media remains unclear. It has been reported that mucin glycoproteins are required for middle ear immune defense against pathogens. We aim to characterize the middle ear effusion (MEE) microbiome using high-throughput sequencing and assess potential associations in microbiome diversity with the presence of the secretory mucins MUC5B and MUC5AC. We hypothesize that MEEs containing MUC5B will exhibit a microbiome largely devoid of typical acute otitis media bacteria. Methods: Fifty-five MEEs from children undergoing myringotomy at Children's National Health System were recovered. Mucin was semiquantitatively determined through Western blot analysis. DNA was subjected to 16S rRNA amplicon sequencing using the Illumina MiSeq platform. Raw data were processed in mothur (SILVA reference database). Alpha- and beta-diversity metrics were determined. Abundance differences between sample groups were estimated. Results: MUC5B was present in 94.5% and MUC5AC in 65.5% of MEEs. Sequencing revealed 39 genera with a relative abundance ≥0.1%. Haemophilus (22.54%), Moraxella (11.11%) and Turicella (7.84%) were the most abundant. Turicella and Pseudomonas proportions were greater in patients older than 24 months of age. In patients with hearing loss, Haemophilus was more abundant, while Turicella and Actinobacteria were less abundant. Haemophilus was also more abundant in samples containing both secretory mucins. Conclusions: The microbiome of MEEs from children with chronic otitis media differs according to specific clinical features, such as mucin content, age and presence of hearing loss. These associations provide novel pathophysiologic insights across the spectrum of otitis media progression.
MeSH term(s)	Adolescent ; Child ; Child, Preschool ; Cohort Studies ; Ear, Middle/microbiology ; Humans ; Infant ; Microbiota/genetics ; Mucin-5B/analysis ; Mucin-5B/metabolism ; Otitis Media with Effusion/epidemiology ; Otitis Media with Effusion/microbiology ; RNA, Ribosomal, 16S/genetics
Chemical Substances	MUC5B protein, human ; Mucin-5B ; RNA, Ribosomal, 16S
Language	English
Publishing date	2016-12-26
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ZDB-ID	392481-6
ISSN	1532-0987 ; 0891-3668
ISSN (online)	1532-0987
ISSN	0891-3668
DOI	10.1097/INF.0000000000001493
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Zs.A 1852: Show issues

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Article ; Online: Proteomic Characterization of Middle Ear Fluid Confirms Neutrophil Extracellular Traps as a Predominant Innate Immune Response in Chronic Otitis Media.

Val, Stephanie / Poley, Marian / Brown, Kristy / Choi, Rachel / Jeong, Stephanie / Colberg-Poley, Annie / Rose, Mary C / Panchapakesan, Karuna C / Devaney, Joe C / Perez-Losada, Marcos / Preciado, Diego

PloS one

2016 Volume 11, Issue 4, Page(s) e0152865

Abstract: Background: Chronic Otitis Media (COM) is characterized by middle ear effusion (MEE) and conductive hearing loss. MEE reflect mucus hypersecretion, but global proteomic profiling of the mucosal components are limited.: Objective: This study aimed at ... ...

Abstract	Background: Chronic Otitis Media (COM) is characterized by middle ear effusion (MEE) and conductive hearing loss. MEE reflect mucus hypersecretion, but global proteomic profiling of the mucosal components are limited. Objective: This study aimed at characterizing the proteome of MEEs from children with COM with the goal of elucidating important innate immune responses. Method: MEEs were collected from children (n = 49) with COM undergoing myringotomy. Mass spectrometry was employed for proteomic profiling in nine samples. Independent samples were further analyzed by cytokine multiplex assay, immunoblotting, neutrophil elastase activity, next generation DNA sequencing, and/or immunofluorescence analysis. Results: 109 unique and common proteins were identified by MS. A majority were innate immune molecules, along with typically intracellular proteins such as histones and actin. 19.5% percent of all mapped peptide counts were from proteins known to be released by neutrophils. Immunofluorescence and immunoblotting demonstrated the presence of neutrophil extracellular traps (NETs) in every MEE, along with MUC5B colocalization. DNA found in effusions revealed unfragmented DNA of human origin. Conclusion: Proteomic analysis of MEEs revealed a predominantly neutrophilic innate mucosal response in which MUC5B is associated with NET DNA. NETs are a primary macromolecular constituent of human COM middle ear effusions.
MeSH term(s)	Child, Preschool ; Chronic Disease ; Extracellular Traps/immunology ; Extracellular Traps/metabolism ; Female ; Humans ; Immunity, Innate ; Infant ; Infant, Newborn ; Male ; Neutrophils/cytology ; Otitis Media with Effusion/immunology ; Otitis Media with Effusion/metabolism ; Proteomics
Language	English
Publishing date	2016-04-14
Publishing country	United States
Document type	Journal Article ; Research Support, N.I.H., Extramural
ISSN	1932-6203
ISSN (online)	1932-6203
DOI	10.1371/journal.pone.0152865
Database	MEDical Literature Analysis and Retrieval System OnLINE

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