scholarly journals Profiling of oral microbiota and cytokines in COVID-19 patients

2020 ◽  
Author(s):  
Valerio Iebba ◽  
Nunzia Zanotta ◽  
Giuseppina Campisciano ◽  
Verena Zerbato ◽  
Stefano Di Bella ◽  
...  

ABSTRACTSARS-CoV-2 presence has been recently demonstrated in the sputum or saliva, suggesting how the shedding of viral RNA outlasts the end of symptoms. Recent data from transcriptome analysis show that oral cavity mucosa harbors high levels of ACE2 and TMPRSS2, highlighting its role as a double-edged sword for SARS-CoV-2 body entrance or interpersonal transmission. In the present study, for the first time, we demonstrate the oral microbiota structure and inflammatory profile of COVID-19 patients. Hospitalized COVID-19 patients and matched healthy controls underwent naso/oral-pharyngeal and oral swabs. Microbiota structure was analyzed by 16S rRNA V2 automated targeted sequencing, while oral and sera concentrations of 27 cytokines were assessed using magnetic bead-based multiplex immunoassays. A significant diminution in species richness was observed in COVID-19 patients, along with a marked difference in beta-diversity. Species such as Prevotella salivae and Veillonella infantium were distinctive for COVID-19 patients, while Neisseria perflava and Granulicatella elegans were predominant in controls. Interestingly, these two groups of oral species oppositely clustered within the bacterial network, defining two distinct Species Interacting Group (SIGs). Pro-inflammatory cytokines were distinctive for COVID-19 in both oral and serum samples, and we found a specific bacterial consortium able to counteract them, following a novel index called C4 firstly proposed here. We even introduced a new parameter, named CytoCOV, able to predict COVID-19 susceptibility for an unknown subject at 71% of power with an AUC equal to 0.995. This pilot study evidenced a distinctive oral microbiota composition in COVID-19 subjects, with a definite structural network in relation to secreted cytokines. Our results would pave the way for a theranostic approach in fighting COVID-19, trying to enlighten the intimate relationship among microbiota and SARS-CoV-2 infection.

2021 ◽  
Vol 12 ◽  
Author(s):  
Valerio Iebba ◽  
Nunzia Zanotta ◽  
Giuseppina Campisciano ◽  
Verena Zerbato ◽  
Stefano Di Bella ◽  
...  

The presence of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has been recently demonstrated in the sputum or saliva, suggesting how the shedding of viral RNA outlasts the end of symptoms. Recent data from transcriptome analysis show that the oral cavity mucosa harbors high levels of angiotensin-converting enzyme 2 (ACE2) and transmembrane protease, serine 2 (TMPRSS2), highlighting its role as a double-edged sword for SARS-CoV-2 body entrance or interpersonal transmission. Here, we studied the oral microbiota structure and inflammatory profile of 26 naive severe coronavirus disease 2019 (COVID-19) patients and 15 controls by 16S rRNA V2 automated targeted sequencing and magnetic bead-based multiplex immunoassays, respectively. A significant diminution in species richness was observed in COVID-19 patients, along with a marked difference in beta-diversity. Species such as Prevotella salivae and Veillonella infantium were distinctive for COVID-19 patients, while Neisseria perflava and Rothia mucilaginosa were predominant in controls. Interestingly, these two groups of oral species oppositely clustered within the bacterial network, defining two distinct Species Interacting Groups (SIGs). COVID-19-related pro-inflammatory cytokines were found in both oral and serum samples, along with a specific bacterial consortium able to counteract them. We introduced a new parameter, named CytoCOV, able to predict COVID-19 susceptibility for an unknown subject at 71% of power with an Area Under Curve (AUC) equal to 0.995. This pilot study evidenced a distinctive oral microbiota composition in COVID-19 subjects, with a definite structural network in relation to secreted cytokines. Our results would be usable in clinics against COVID-19, using bacterial consortia as biomarkers or to reduce local inflammation.


2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 469-470
Author(s):  
M. Luque-Tévar ◽  
C. Perez-Sanchez ◽  
N. Barbarroja Puerto ◽  
A. M. Patiño-Trives ◽  
I. Arias de la Rosa ◽  
...  

Background:TNF inhibitors (TNFi) represent an extraordinary advance in the management of Rheumatoid Arthritis (RA). Despite their benefits, there is a percentage of patients (20–40%) that do not achieve clinical improvement. Therefore, it is necessary to search for new and easily accessible biomarkers predictive of therapeutic response that might guide precision medicine.Objectives:1. To explore changes in the molecular profile of RA patients following TNFi therapy in serum samples. 2. To search for new and reliable biomarkers predictive of TNFi response, based on clinical and molecular profiles of RA patients, by using machine learning algorithms.Methods:In a prospective multicenter study, 79 RA patients undergoing TNFi and 29 healthy donors (HD) were enrolled. Twenty-two RA patients were further included as a validation cohort. Serum samples were obtained before and after 6 months of treatment, and therapeutic efficacy was evaluated. Patients’ response was determined following EULAR response criteria. Serum inflammatory profile was analyzed by a multiplex immunoassay, along with oxidative and NETotic profiles, evaluated by commercial kits. A circulating miRNA array was also performed by next-generation sequencing. Clustering analysis was carried out to identify groups of patients with distinctive molecular signatures. Then, clinical and molecular changes induced by TNFi were delineated after 6 months of therapy. Finally, integrative clinical and molecular signatures as predictors of response were assessed at baseline by supervised machine learning methods, using regularized logistic regressions.Results:Inflammatory, oxidative stress and NETosis-derived biomolecules were found altered in RA patients versus HD, closely interconnected and associated with several deregulated miRNAs. This altered molecular profile at baseline allowed the unsupervised division of three clusters of RA patients with distinctive clinical phenotypes, further linked to TNFi effectiveness. Cluster 1 included RA patients with low levels of pro-inflammatory cytokines, associated with a medium-low disease activity score and good clinical response. Clusters 2-3 comprised patients with high levels of pro-inflammatory cytokines, associated with a high disease activity and a non-response rate of 30%.After 6 months of therapy the molecular profile found altered in RA patients was reversed in responder patients, who achieved a molecular phenotype similar to HDs. However, non-responder patients’ molecular profile remained significantly deregulated, including alterations in inflammatory mediators (IL-6, L-8, TNFα, VEGF, IL-1RA, IL-5, IL-15, GMCSF, GCSF, FGFb), oxidative stress markers (LPO) and NETosis-derived products (Elastase), along with specific miRNAs (miR-199a-5p). These molecular changes further correlated with changes in disease activity score. Machine-learning algorithms identified clinical (Creatinine, IgM, Vitamin D, Swollen Joints, C4, Disease Duration and Tryglicerides) and molecular (Nucleosomes, IL-10, miR-106a-5p, IL-13, IL-12p70, IL-15 and LPO) signatures as potential predictors of response to TNFi treatment with high accuracy. Furthermore, the integration of both features in a combined model increased the predictive value of these signatures (AUC: 0.91). These results were further confirmed in an independent validation cohort.Conclusion:1. RA patients display distinctive altered molecular profiles directly linked to their clinical status and associated with TNFi effectiveness. 2. Clinical response was associated with a specific modulation of the inflammatory profile, the reestablishment of the altered oxidative status, the reduction of NETosis and the reversion of related altered miRNAs. 3. The integrative analysis of the clinical and molecular profiles using machine learning allows the identification of novel signatures as potential predictors of therapeutic response to TNFi therapy.Disclosure of Interests:None declared


Tick-borne encephalitis virus (TBEV) was isolated for the first time in Sweden in 1958 (from ticks and from 1 tick-borne encephalitis [TBE] patient).1 In 2003, Haglund and colleagues reported the isolation and antigenic and genetic characterization of 14 TBEV strains from Swedish patients (samples collected 1991–1994).2 The first serum sample, from which TBEV was isolated, was obtained 2–10 days after onset of disease and found to be negative for anti-TBEV immunoglobulin M (IgM) by enzyme-linked immunosorbent assay (ELISA), whereas TBEV-specific IgM (and TBEV-specific immunoglobulin G/cerebrospinal fluid [IgG/CSF] activity) was demonstrated in later serum samples taken during the second phase of the disease.


2019 ◽  
Vol 71 ((suppl.1)) ◽  
pp. 209-243
Author(s):  
J.K.H. Koh ◽  
D.J. Court

This paper discusses the preliminary results of the first comprehensive survey of the spiders of the Bukit Timah Nature Reserve (BTNR) in Singapore. Two plots were established in each of the three zones of vegetation, viz., primary forest, old secondary forest, and maturing secondary forest. They were repeatedly sampled over an 18-month period. Sorting of the collection so far suggests that the three vegetation zones harbour rather different spider assemblages. Only ~9% of the total spider fauna recovered was shared by all three zones. The results have also yielded a preliminary picture of dominance, abundance and rarity. Although first intended to obtain a baseline for future quantitative analyses, the survey became a testing ground to modify and refine methodology so as to conduct future quantitative surveys with greater scientific rigour. Taxonomic work on the samples so far shows that the spiders in the BTNR span over 43 families, of which six families are listed for the first time in Singapore. The tally is summarised in an interim checklist of BTNR spiders. The checklist, with a total of 317 entries, shows that there are 158 described species of spiders in BTNR, of which 25 species are new records for Singapore. Another 159 morphospecies are provisionally recognised as distinct species, some of which may be new to science. Our observations during the survey have allowed us to provide a narrative of BTNR spider diversity against a backdrop of their microhabitat specialisation.


2016 ◽  
Vol 1 (1) ◽  
Author(s):  
D.G. Shah ◽  
D.N. Mehta ◽  
R.V. Gujar

Bryophytes are the second largest group of land plants and are also known as the amphibians of the plant kingdom. 67 species of bryophytes have been reported from select locations across the state of Gujrat. The status of family fissidentaceae which is a large moss family is being presented in this paper. Globally the family consists of 10 genera but only one genus, Fissidens Hedw. has been collected from Gujarat. Fissidens is characterized by a unique leaf structure and shows the presence of three distinct lamina, the dorsal, the ventral and the vaginant lamina. A total of 8 species of Fissidens have been reported from the state based on vegetative characters as no sporophyte stages were collected earlier. Species reported from the neighboring states also showed the absence of sporophytes. The identification of different species was difficult due to substantial overlap in vegetative characters. Hence a detailed study on the diversity of members of Fissidentaceae in Gujarat was carried out between November 2013 and February 2015. In present study 8 distinct species of Fissidens have been collected from different parts of the state. Three species Fissidens splachnobryoides Broth., Fissidens zollingerii Mont. and Fissidens curvato-involutus Dixon. have been identified while the other five are still to be identified. Fissidens zollingerii Mont. and Fissidens xiphoides M. Fleisch., which have been reported as distinct species are actually synonyms according to TROPICOS database. The presence of sexual reproductive structures and sporophytes for several Fissidens species are also being reported for the first time from the state.


Pathogens ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 725
Author(s):  
David Becerro-Recio ◽  
Javier González-Miguel ◽  
Alberto Ucero ◽  
Javier Sotillo ◽  
Álvaro Martínez-Moreno ◽  
...  

Excretory/secretory products released by helminth parasites have been widely studied for their diagnostic utility, immunomodulatory properties, as well as for their use as vaccines. Due to their location at the host/parasite interface, the characterization of parasite secretions is important to unravel the molecular interactions governing the relationships between helminth parasites and their hosts. In this study, the excretory/secretory products from adult worms of the trematode Fasciola hepatica (FhES) were employed in a combination of two-dimensional electrophoresis, immunoblot and mass spectrometry, to analyze the immune response elicited in sheep during the course of an experimental infection. Ten different immunogenic proteins from FhES recognized by serum samples from infected sheep at 4, 8, and/or 12 weeks post-infection were identified. Among these, different isoforms of cathepsin L and B, peroxiredoxin, calmodulin, or glutathione S-transferase were recognized from the beginning to the end of the experimental infection, suggesting their potential role as immunomodulatory antigens. Furthermore, four FhES proteins (C2H2-type domain-containing protein, ferritin, superoxide dismutase, and globin-3) were identified for the first time as non-immunogenic proteins. These results may help to further understand host/parasite relationships in fasciolosis, and to identify potential diagnostic molecules and drug target candidates of F. hepatica.


2021 ◽  
Vol 18 ◽  
Author(s):  
Jamila Tabassum ◽  
Muhammad Luqman ◽  
Fariha Hasan ◽  
Fariha Arooj ◽  
Saif Ur Rehman Kashif ◽  
...  

: Synthetic plastic waste management is a tenacious environmental concern at the global level. Although all types of synthetic plastics are a nuisance to the environment, however, versatility and one time use have made polyethylene (PE) a foremost environmental issue. The current study has investigated cavern bacterial strains isolated from PE samples from San Giovanni cave, Sardinia, Italy for their efficacy to biodegrade low-density polyethylene (LDPE) film. It was an initial effort to use cavern bacteria in plastic biodegradation studies. Chemical and physical changes in the composition of LDPE were studied by Fourier-transform infrared spectroscopy (FTIR) and scanning electron microscopy (SEM) after incubation with the bacterial consortium for two months. Collected cavern PE plastic samples were also studied for biodegradation after incubation in nutrient broth for two months. FTIR revealed obvious signs of degradation with the appearance of two new peaks of functional groups, nitriles (C≡N) and amines (N-H) in LDPE film, which are intermediate metabolites of β- oxidation pathway. An increase in various existing peaks of several intermediate metabolites including aldehydes, ketones, alcohols, and carboxylic acids, were also observed in experimental LDPE compared to control. Peaks of alkanes decreased significantly owing to cavern bacterial activity. SEM revealed biofilm formation on experimental LDPE surface with substantial mechanical damage. Similar signs of degradation were observed in the cavern PE samples. Four bacterial strains in the current consortium, including Bacillus sonorensis, Bacillus subtilis, Aneurinibacillus spp., and Alcaligenes faecalis are first time reported to be linked with biodegradation of plastics. The cavern bacteria under study have the potential to biodegrade LDPE.


2017 ◽  
Vol 11 (11) ◽  
pp. 854-860 ◽  
Author(s):  
Ahmed A Zaghawa ◽  
Fadhel Housawi ◽  
Abdulmohsen Al-Naeem ◽  
Ahmed Elsify ◽  
Yamen Mohammed Hegazy

Introduction: Bovine ephemeral fever virus (BEFV) is an arthropod borne Rhabdovirus affects cattle and water buffalo causes acute febrile disease. Methodology: The clinical picture and epidemiological pattern of BEF were described among cattle in epidemics of 2007, 2009 and 2011 in four geographical regions of Kingdom Saudi Arabia (Eastern, Jizan, Qasim, and Riyadh). Serum samples were tested using VNT. Virus isolation and molecular characterization were carried out for the first time in KSA. Results: The main clinical symptoms were fever, stiffness, lameness, salivation and subcutaneous emphysema. The prevalence and the mortality rate of BEF have decreased from 70% and 4.6% in 2007 to 30% and 0.6% in 2011, respectively in the 4 studied areas. There was no region association with higher prevalence of BEF. The intracluster correlation (ICC) was estimated for the first time in KSA as 0.0034. BEFV had been isolated from 11 out of 20 samples (55%) and isolation was confirmed by VNT. The molecular detection of BEFV by RT-PCR and real- time RT-qPCR were found more sensitive for diagnosis of the disease than virus isolation; 80% and 90% for the former tests and 55% for the latter. Three isolates were sequenced, they showed 84.7% - 100% identities in between and shared 90.4%-96.5% sequence identity with a previously published sequence from Australia (KF679404). The generated sequences belonged to 3rd cluster of BEFV glycoprotein. Conclusions: BEF occurrence has cyclic nature and the efficacy of vaccines prepared from local strains has to be evaluated and considered in diseases control.


2016 ◽  
Vol 85 (1) ◽  
pp. 71-76 ◽  
Author(s):  
Sevim Kasap ◽  
Hasan Batmaz ◽  
Meric Kocaturk ◽  
Frank Gessler ◽  
Serkan Catık ◽  
...  

This paper presents the case of a six year-old, male, thoroughbred horse with clinical signs of inappetence, weakness, and incoordination when walking. Clinical examination showed that the horse staggered and leaned to the left side. Feedstuff was present inside and around its mouth. Salivation was increased and there was no reflex at the palpebrae and tongue. The horse had difficulty swallowing and the tone of its tail was reduced. Botulism was diagnosed based on the clinical signs. Antibiotic (ceftiofur) and fluid-electrolyte treatment was commenced. Next day, neostigmin was added to the horse’s treatment, and it became recumbent. The horse’s palpebral, tongue and tail reflexes returned partially after neostigmine methylsulphate treatment on the same day and it stood up on day four. However, it could not swallow anything during the whole week, so after getting permission from the owner, the horse was euthanized on day 10. Samples of the colonic content and blood serum were sent by courier to the laboratory for toxin neutralization, however, botulinum neurotoxins could not be detected. After that, serum samples from days 6 and 10 were sent to another laboratory for testing for botulinum neurotoxin antibodies by ELISA. Specific antibodies against botulinum neurotoxin type A were measured, indicating a previous, immuno-relevant contact with the toxin. This seroconversion for type A supports the clinical botulism diagnosis. Type A botulism is rarely seen in Europe and has been detected in a horse in Turkey for the first time.


2021 ◽  
Author(s):  
Huma Jawed ◽  
Syed Uzair Ali Shah ◽  
Shahid I. Awan ◽  
Shazia Anjum ◽  
Shabana U. Simjee

ABSTRACTThe anti-inflammatory and anti-arthritic activity of GN1 (6-hydroxymethyl-3-(1-methylene-allylamino)-tetrahydro-pyran-2,4,5-triol), an analog of glucoseamine was investigated in the adjuvant induced arthritis (AIA) in rats. It was observed that the highly increased hind paw swelling was significantly reduced (p <0.001) with no noticeable retardation of body weight in the GN1 treated arthritic rats as compared to arthritic control rats. The histopathological analysis of isolated rat joints by scaling system classification of different stages appear in arthritic inflammatory lesion development, disclosed the suppressive effect in the inflammation processing in the GN1 treated animals. Different predictive markers such as TNF-α, nitric oxide IL-1β, peroxide and glutathione were also determined in the serum samples of the treated and non-treated arthritic rats in order to measure the intensity of inflammation. Our results show that GN-1 treatment in arthritic groups demonstrated marked reduction in c-fos expression. Results also revealed that GN1 significantly reduces the peroxide production (p < 0.002) and both the pro-inflammatory cytokines TNF-α (p < 0.015) and IL-1β (p < 0.001) in the arthritic rats receiving this treatment. Although, nitric oxide formation was found to be less than the arthritic control, but this reduction was not significant. The level of glutathione was significantly increased in GN1 treated arthritic rats (p < 0.037). These observations suggest that GN1 may help in reducing the inflammation and have anti-arthritic properties.


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