Role of Anaerobes in Chronic Sinusitis: Will Polymerase Chain Reaction Solve the Debate

2002 ◽  
Vol 127 (5) ◽  
pp. 384-386 ◽  
Author(s):  
Hassan H. Ramadan ◽  
Peter H. Mathers ◽  
Heather Schwartzbauer
Keyword(s):  
Chronic Rhinosinusitis ◽  
Chronic Sinusitis ◽  
Anaerobic Bacteria ◽  
Sinus Surgery ◽  
Pcr Analysis ◽  
Chain Reaction ◽  
Culture Techniques ◽  
Standard Culture ◽  
Polymerase Chain

BACKGROUND: Bacteriology of chronic sinusitis continues to be a matter of debate, particularly the role of anaerobes. Some authors suggest that anaerobes play a significant role whereas others suggest a minimal role. Those who suggest a significant role argue that standard culture techniques are the culprits. OBJECTIVE: The purpose of this study was to use polymerase chain reaction (PCR) on sinus specimens for the presence of anaerobes and to compare them with standard culture techniques. METHODS: Sixty-four samples were obtained in a sterile fashion during sinus surgery and were sent for standard anaerobic cultures as well as anaerobic PCR analysis. RESULTS: Anaerobic bacteria were demonstrated in 5% of culture specimens, which is similar to recently published data. PCR identified anaerobic bacteria in 19% of the specimens ( P = 0.01) CONCLUSION: PCR analysis of surgical samples obtained during endoscopic sinus surgery for chronic sinusitis identified a significantly higher incidence of anaerobes (x4) compared with standard anaerobic culture technique. Chronic rhinosinusitis is one of the most common chronic diseases that affects individuals in the United States. It is estimated that >25 million office visits are made for sinusitis. 1 The financial impact of chronic sinusitis includes not only the direct medical costs of treatment but also the millions of hours of lost productivity caused by the disease. 1 Chronic rhinosinusitis is defined as the signs and symptoms of sinus inflammation that last longer than 12 weeks associated with documented sinus inflammation with imaging techniques ≥4 weeks after appropriate antibiotic therapy. 2 There is agreement in the literature regarding the bacterial etiology of acute rhinosinusitis. However, the etiology of chronic rhinosinusitis is still unclear despite numerous articles about the bacteriology of chronic rhinosinusitis. 3–10 The role of aerobes is more clear than the role of anaerobes in chronic rhinosinusitis; many conflicting reports have been published about the role of anaerobes as etiologic factors in chronic rhinosinusitis. 3–10 The incidence of anaerobes obtained on standard cultures from patients with chronic rhinosinusitis ranges from 0% to 56%. 3–10 The reports that show a high yield of anaerobes argue that the technique used to obtain the specimens, the method of transportation, and even specimen collection are reasons why other reports did not yield high levels of anaerobes. 4–5 With increasing numbers of patients with chronic rhinosinusitis and with increasing bacterial resistance to antimicrobials, which is blamed on the improper use of antimicrobials, knowledge of the bacteriology becomes important in the treatment. The role of aerobes was addressed in a prior publication 11 ; the role of anaerobic bacteria as an etiologic cause of chronic rhinosinusitis is addressed in this report. The goal of this study was to test specimens, using 2 different techniques, obtained during endoscopic sinus surgery from individuals who had chronic rhinosinusitis.

Distinguishing Molecular Features of Allergic Fungal Rhinosinusitis

2018 ◽  
Vol 159 (1) ◽  
pp. 185-193 ◽  
Author(s):  
Matthew A. Tyler ◽  
Caroline J. Padro Dietz ◽  
Chris B. Russell ◽  
Martin J. Citardi ◽  
Shervin Assassi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Rhinosinusitis ◽  
Quantitative Polymerase Chain Reaction ◽  
Sinus Surgery ◽  
Type I ◽  
Chain Reaction ◽  
Molecular Features ◽  
Fungal Rhinosinusitis ◽  
Allergic Fungal Rhinosinusitis ◽  
Polymerase Chain

Objective Allergic fungal rhinosinusitis (AFRS) is a clinical subtype of chronic rhinosinusitis with nasal polyps (CRSwNP), characterized by eosinophilic mucin, evidence of fungal elements within the mucin, fungal-specific type I hypersensitivity, and characteristic computed tomography findings. It remains controversial whether AFRS represents a disease with a unique pathophysiology from chronic rhinosinusitis or is merely a severe form of CRSwNP. The goal of this study was to identify molecular features unique to AFRS. Study Design Cross-sectional case-control. Setting Single academic tertiary referral institution. Subjects and Methods Subjects included 86 patients undergoing endoscopic sinus surgery: CRSwNP (n = 34), AFRS (n = 37), and healthy controls (n = 15). Pathway and correlation analyses were performed with whole-genome microarray data for study patients undergoing surgery for recalcitrant chronic rhinosinusitis. Our findings were confirmed with quantitative polymerase chain reaction and immunohistochemical studies. Results AFRS was uniquely characterized by a pronounced association with adaptive T helper 2–associated immune gene expression. AFRS exhibited altered expression of proteins associated with secretory salivary peptides—namely, histatin, a peptide with known antifungal activity in the oral cavity. Furthermore, the expression of histatins correlated negatively with that of type 2 inflammatory mediators. We confirm the decreased expression of histatins in AFRS when compared with CRSwNP by quantitative polymerase chain reaction and localized its expression to a submucosal cell population. Conclusion There exist clear molecular profiles that distinguish AFRS from CRSwNP. This divergence translates into an altered ability to control fungal growth and may in part explain some of the phenotypical differences between CRSwNP and AFRS.

Mycoplasma pneumoniaeandChlamydophila pneumoniae: a comparative study in patients with nasal polyposis and healthy controls

2015 ◽  
Vol 129 (9) ◽  
pp. 865-869 ◽  
Author(s):  
D G Ioannidis ◽  
V A Lachanas ◽  
Z Florou ◽  
J G Bizakis ◽  
E Petinaki ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Chronic Rhinosinusitis ◽  
Nasal Polyps ◽  
Inferior Turbinate ◽  
Sinus Surgery ◽  
Control Group ◽  
Chain Reaction ◽  
Tissue Samples ◽  
Polymerase Chain

AbstractIntroduction:The role played byMycoplasma pneumoniaeandChlamydophila pneumoniaein the pathogenesis of chronic rhinosinusitis with nasal polyps has been the object of ongoing debate. We used real-time polymerase chain reaction to investigate the prevalence of both microorganisms in the nasal tissue samples of patients and controls.Methods:We extracted DNA from nasal polyp samples obtained during functional endoscopic sinus surgery and the inferior turbinate samples of controls undergoing septoplasty. We used the highly sensitive real-time polymerase chain reaction to detect the presence ofM pneumoniaeandC pneumoniaeDNA.Results:Patients with chronic rhinosinusitis with nasal polyps consisted of 62 individuals (39 men; mean age 51 years); the control group consisted of 24 individuals (13 men; mean age 45 years). All samples from both groups were negative forM pneumoniaeandC pneumoniaeDNA.Conclusion:We have demonstrated that the likelihood ofM pneumoniaeandC pneumoniaeacting as an ongoing inflammatory stimulus in chronic rhinosinusitis with nasal polyps is slim.

scholarly journals Downregulated MiR-206 expression promotes the proliferation and migration of macrophages by regulating IL-17A/REG3A pathway

2020 ◽  
Vol 18 ◽  
pp. 205873922091749
Author(s):  
Xuan Huang ◽  
Fang Gong ◽  
Zhixian Lu ◽  
Jie Zhu ◽  
Qun Yu
Keyword(s):  
Pcr Analysis ◽  
Chain Reaction ◽  
Inflammatory Infiltration ◽  
Interleukin 17A ◽  
Polymerase Chain ◽  
And Migration ◽  
Cell Migration And Proliferation ◽  
Migration Of Macrophages ◽  
Proliferation And Migration

This study sought to investigate the role of miR-206 in polymyositis/dermatomyositis (PM/DM) development. Transwell and 3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium inner salt (MTS) assay were performed to investigate cell migration and proliferation. Real-time polymerase chain reaction (PCR) analysis was used to determine the expression of miR-206, interleukin-17A (IL-17A), IL-17 receptor A (IL-17RA), and regenerating islet-derived protein 3-alpha (REG3A). Significantly, miR-206 mimics decreased macrophage migration and proliferation, while miR-206 inhibitors exhibited the opposite effects. Indeed, elevating IL-17RA levels resulted in increased REG3A expression, which was inhibited by IL-17RA siRNA. Besides, miR-206 mimics decreased IL-17A and REG3A expressions, but miR-206 inhibitors showed opposite effects. Moreover, miR-206 expression in PM/DM patients was significantly reduced compared with the healthy controls, while IL-17A and REG3A expressions substantially increased among PM/DM patients. These findings suggested that downregulation of miR-206 increased the migration and proliferation of macrophages via IL-17A/REG3A signaling pathway, which could promote the inflammatory infiltration in PM/DM.

scholarly journals Identification of uncultured bacteria from abscesses of exotic pet animals using broad-range nested 16S rRNA polymerase chain reaction and Sanger sequencing

2019 ◽  
Vol 12 (10) ◽  
pp. 1546-1553
Author(s):  
T. Duangurai ◽  
J. Siengsanan-Lamont ◽  
C. Bumrungpun ◽  
G. Kaewmongkol ◽  
L. Areevijittrakul ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
16S Rrna ◽  
Sanger Sequencing ◽  
Anaerobic Bacteria ◽  
Rpob Gene ◽  
Chain Reaction ◽  
Sequencing Technique ◽  
Standard Culture ◽  
Polymerase Chain ◽  
Pcr Techniques

Background: The Sanger sequencing technique has been questioned and challenged by advanced high-throughput sequencing approaches. Sanger sequencing seems to be an obsolete technology. However, there are still research problems that could be answered using the Sanger sequencing technology. Fastidious obligate anaerobic bacteria are mostly associated with abscesses in animals. These bacteria are difficult to isolate from abscesses and are frequently excluded due to the bias of conventional bacterial culturing. Aim: This study demonstrated the usefulness of a broad-range polymerase chain reaction (PCR) with Sanger sequencing to identify the majority population of bacteria in abscesses from exotic pet animals. Materials and Methods: This study performed a pilot investigation of abscesses from 20 clinical cases (17 rabbits, 2 hedgehogs, and 1 sugar glider) using standard culture methods for both aerobes and anaerobes and broad-range nested PCR targeting the 16S rRNA gene followed by the Sanger sequencing technique. Results: The standard culture and PCR techniques detected bacteria in 9 and 17 of 20 samples, respectively. From the 17 sequencings of the 16S rRNA, 10 PCR products were found to be closely related with obligate anaerobes including Bacteroides spp., Fusobacterium spp., Prevotella spp. Phylogenetic analysis using the rpoB gene revealed that the species for the Bacteroides was thetaiotaomicron and for the Fusobacterium was varium and nucleatum. However, the amplification of the rpoB gene for the Prevotella spp. was unsuccessful. Correlations between the standard culture and PCR techniques were found in 9 (6 positive and 3 negative samples) of 20 samples. Eleven samples were discordant between the standard culture and PCR techniques which were composed of eight samples negative by culture but positive by PCR and three samples had different bacteria by the culture and PCR techniques. Conclusion: According to this study, broad-range PCR combined with Sanger sequencing might be useful for the detection of dominant anaerobic bacteria in abscesses that were overlooked based on conventional bacterial culture.

scholarly journals Association Between Culture and Culture-Independent Microtyping in Recalcitrant Chronic Rhinosinusitis

2019 ◽  
Vol 98 (2) ◽  
pp. 94-97 ◽  
Author(s):  
Andrew Vaughn ◽  
Courtney Shaver ◽  
David Clark
Keyword(s):  
Polymerase Chain Reaction ◽  
Chronic Rhinosinusitis ◽  
Medical Records ◽  
Sinus Surgery ◽  
Chain Reaction ◽  
Persistent Symptoms ◽  
Culture Independent ◽  
Polymerase Chain ◽  
Bacteria And Fungi ◽  
Culture Dependent

Background: Many different etiologies have been proposed to be responsible for the pathogenesis of chronic rhinosinusitis, including dysbiosis of the sinus microbiome. Attempts have recently been made to identify a pathogenic organism via advanced culture mechanisms. The purpose of this study is to use culture-dependent and culture-independent means of microtyping to determine whether any association exists between the quantity and quality of bacteria identified in patients with recalcitrant chronic rhinosinusitis. Methods: Medical records were retrospectively reviewed for patients with a history of revision sinus surgery and persistent symptoms who underwent endoscopically directed culture and underwent quantitative polymerase chain reaction analysis of the 16S ribosomal RNA of bacteria and fungi from February 1, 2014, to January 1, 2017. A total of 21 patients met the inclusion criteria. Medical records were reviewed to determine the number of bacterial isolates and relative abundance of bacteria and fungi on culture and polymerase chain reaction. Results: Using culture-independent techniques of examining purulent secretions in patients with recalcitrant chronic rhinosinusitis, an average of 3.61 isolates were identified per specimen, compared with culture-dependent methods that revealed 2.10 isolates per specimen ( P < .05). The dominant species identified on each culture was rarely the most abundant species identified using polymerase chain reaction techniques. Conclusions: Traditional culture methodologies may fail to identify potential pathogens or the dominant pathogen in patients with recalcitrant chronic rhinosinusitis with acute exacerbations.

Sensitivity of polymerase chain reaction (PCR)-southern hybridization and conventional PCR analysis for Halal authentication of gelatin capsules

LWT ◽  
2015 ◽  
Vol 63 (1) ◽  
pp. 714-719 ◽  
Author(s):  
Sahilah Abd Mutalib ◽  
Nursheila Mustafa Muin ◽  
Aminah Abdullah ◽  
Osman Hassan ◽  
Wan Aida Wan Mustapha ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Southern Hybridization ◽  
Pcr Analysis ◽  
Conventional Pcr ◽  
Chain Reaction ◽  
Gelatin Capsules ◽  
Halal Authentication ◽  
Polymerase Chain

scholarly journals Decreased Expression of GPER1 Gene and Protein in Goiter

2015 ◽  
Vol 2015 ◽  
pp. 1-5 ◽  
Author(s):  
Raquel Weber ◽  
Ana Paula Santin Bertoni ◽  
Laura Walter Bessestil ◽  
Ilma Simoni Brum ◽  
Tania Weber Furlanetto
Keyword(s):  
Estrogen Receptor ◽  
Quantitative Polymerase Chain Reaction ◽  
Thyroid Tissue ◽  
Chain Reaction ◽  
Normal Thyroid ◽  
Protein Levels ◽  
Protein Expressions ◽  
Polymerase Chain ◽  
G Protein Coupled

Goiter is more common in women, suggesting that estrogen could be involved in its physiopathology. The presence of classical estrogen receptors (ERαand ERβ) has been described in thyroid tissue, suggesting a direct effect of estrogen on the gland. A nonclassic estrogen receptor, the G-protein-coupled estrogen receptor (GPER1), has been described recently in several tissues. However, in goiter, the presence of this receptor has not been studied yet. We investigated GPER1 gene and protein expressions in normal thyroid and goiter using reverse transcription quantitative polymerase chain reaction (RT-qPCR) and Western blot, respectively. In normal thyroid (n=16) and goiter (n=19), GPER1 gene was expressed in all samples, while GPER1 protein was expressed in all samples of normal thyroid (n=15) but in only 72% of goiter samples (n=13). When comparing GPER1 gene and protein levels in both conditions, gene expression and protein levels were higher in normal thyroid than in goiter, suggesting a role of this receptor in this condition. Further studies are needed to elucidate the role of GPER1 in normal thyroid and goiter.

scholarly journals Macrophage migration inhibitory factor may play a protective role in osteoarthritis

2021 ◽  
Vol 23 (1) ◽  
Author(s):  
Ming Liu ◽  
Zikun Xie ◽  
Guang Sun ◽  
Liujun Chen ◽  
Dake Qi ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Migration Inhibitory Factor ◽  
Macrophage Migration Inhibitory Factor ◽  
Protective Role ◽  
Macrophage Migration ◽  
Chain Reaction ◽  
Protein Levels ◽  
Tnf Α ◽  
Polymerase Chain

Abstract Background Osteoarthritis (OA) is the most prevalent form of arthritis and the major cause of disability and overall diminution of quality of life in the elderly population. Currently there is no cure for OA, partly due to the large gaps in our understanding of its underlying molecular and cellular mechanisms. Macrophage migration inhibitory factor (MIF) is a procytokine that mediates pleiotropic inflammatory effects in inflammatory diseases such as rheumatoid arthritis (RA) and ankylosing spondylitis (AS). However, data on the role of MIF in OA is limited with conflicting results. We undertook this study to investigate the role of MIF in OA by examining MIF genotype, mRNA expression, and protein levels in the Newfoundland Osteoarthritis Study. Methods One hundred nineteen end-stage knee/hip OA patients, 16 RA patients, and 113 healthy controls were included in the study. Two polymorphisms in the MIF gene, rs755622, and -794 CATT5-8, were genotyped using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) and PCR followed by automated capillary electrophoresis, respectively. MIF mRNA levels in articular cartilage and subchondral bone were measured by quantitative polymerase chain reaction. Plasma concentrations of MIF, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and interleukin-1 beta (IL-1β) were measured by enzyme-linked immunosorbent assay. Results rs755622 and -794 CATT5-8 genotypes were not associated with MIF mRNA or protein levels or OA (all p ≥ 0.19). MIF mRNA level in cartilage was lower in OA patients than in controls (p = 0.028) and RA patients (p = 0.004), while the levels in bone were comparable between OA patients and controls (p = 0.165). MIF protein level in plasma was lower in OA patients than in controls (p = 3.01 × 10−10), while the levels of TNF-α, IL-6 and IL-1β in plasma were all significantly higher in OA patients than in controls (all p ≤ 0.0007). Multivariable logistic regression showed lower MIF and higher IL-1β protein levels in plasma were independently associated with OA (OR per SD increase = 0.10 and 8.08; 95% CI = 0.04–0.19 and 4.42–16.82, respectively), but TNF-α and IL-6 became non-significant. Conclusions Reduced MIF mRNA and protein expression in OA patients suggested MIF might have a protective role in OA and could serve as a biomarker to differentiate OA from other joint disorders.

Development of Three-Stage Bioaerosol Sampler for Size-Selective Sampling

2022 ◽  
Author(s):  
Jun-Hyung Lim ◽  
Sang Hwan Nam ◽  
Jongwoo Kim ◽  
Nam Hoon Kim ◽  
Gun-Soo Park ◽  
...  
Keyword(s):  
Polymerase Chain Reaction ◽  
Size Range ◽  
Collection Efficiency ◽  
Commercial Product ◽  
Pcr Analysis ◽  
Cyclone Separator ◽  
Chain Reaction ◽  
Selective Sampling ◽  
Sampling Flow Rate ◽  
Polymerase Chain

Abstract In this study, a three-stage bioaerosol sampler with a sampling flow rate of 170 L/min was designed and fabricated for sampling the bioaerosols released during human breathing and coughing, and its performance was evaluated. The sampler was constructed using a cyclone separator with a cutoff size of 2.5 µm as a preseparator, a multi-nozzle virtual impactor with a cutoff size of 0.34 µm as an aerosol concentrator, and a BioSampler, which is a commercial product, for collecting bioaerosols in a collection fluid. The collection efficiency of the sampler was evaluated through simulations and experiments. Only particles with sizes of 0.1-4 µm were selectively collected in the collection fluid. Bacteriophage bioaerosols were sampled using the developed sampler and ACD-200 Bobcat sampler, which is a commercial product. The amounts of collected bacteriophages were compared using the polymerase chain reaction (PCR) technique. The sampling performance of the developed sampler was similar to that of the ACD-200 Bobcat sampler. Moreover, the developed sampler showed its ability to sample bioaerosols of a specific size-range and collect them directly in a collection fluid for the PCR analysis. Therefore, the developed sampler is expected to be useful for indoor environmental monitoring by effectively sampling the bioaerosols released indoors during human breathing and coughing.

Sign in / Sign up

Export Citation Format

Share Document