scholarly journals Culturable and Non-Culturable Blood Microbiota of Healthy Individuals

2021 ◽  
Vol 9 (7) ◽  
pp. 1464
Author(s):  
Stefan Panaiotov ◽  
Yordan Hodzhev ◽  
Borislava Tsafarova ◽  
Vladimir Tolchkov ◽  
Reni Kalfin
Keyword(s):  
Fungal Species ◽  
Blood Collection ◽  
Healthy Individuals ◽  
High Concentration ◽  
Complex Samples ◽  
Bacterial Phyla ◽  
A Cell ◽  
Free Circulating Dna ◽  
Culturing Conditions ◽  
Blood Microbiota

Next-generation sequencing (NGS) and metagenomics revolutionized our capacity for analysis and identification of the microbial communities in complex samples. The existence of a blood microbiome in healthy individuals has been confirmed by sequencing, but some researchers suspect that this is a cell-free circulating DNA in blood, while others have had isolated a limited number of bacterial and fungal species by culture. It is not clear what part of the blood microbiota could be resuscitated and cultured. Here, we quantitatively measured the culturable part of blood microbiota of healthy individuals by testing a medium supplemented with a high concentration of vitamin K (1 mg/mL) and culturing at 43 °C for 24 h. We applied targeted sequencing of 16S rDNA and internal transcribed spacer (ITS) markers on cultured and non-cultured blood samples from 28 healthy individuals. Dominant bacterial phyla among non-cultured samples were Proteobacteria 92.97%, Firmicutes 2.18%, Actinobacteria 1.74% and Planctomycetes 1.55%, while among cultured samples Proteobacteria were 47.83%, Firmicutes 25.85%, Actinobacteria 16.42%, Bacteroidetes 3.48%, Cyanobacteria 2.74%, and Fusobacteria 1.53%. Fungi phyla Basidiomycota, Ascomycota, and unidentified fungi were 65.08%, 17.72%, and 17.2% respectively among non-cultured samples, while among cultured samples they were 58.08%, 21.72%, and 20.2% respectively. In cultured and non-cultured samples we identified 241 OTUs belonging to 40 bacterial orders comprising 66 families and 105 genera. Fungal biodiversity accounted for 272 OTUs distributed in 61 orders, 105 families, and 133 genera. Bacterial orders that remained non-cultured, compared to blood microbiota isolated from fresh blood collection, were Sphingomonadales, Rhizobiales, and Rhodospirillales. Species of orders Bacillales, Lactobacillales, and Corynebacteriales showed the best cultivability. Fungi orders Tremellales, Polyporales, and Filobasidiales were mostly unculturable. Species of fungi orders Pleosporales, Saccharomycetales, and Helotiales were among the culturable ones. In this study, we quantified the capacity of a specific medium applied for culturing of blood microbiota in healthy individuals. Other culturing conditions and media should be tested for optimization and better characterization of blood microbiota in healthy and diseased individuals.

scholarly journals I Like the Way You Eat It: Lemur (Indri indri) Gut Mycobiome and Geophagy

2021 ◽  
Author(s):  
Luigimaria Borruso ◽  
Alice Checcucci ◽  
Valeria Torti ◽  
Federico Correa ◽  
Camillo Sandri ◽  
...  
Keyword(s):  
Plant Material ◽  
Soil Characteristics ◽  
Nutrient Supply ◽  
Fungal Species ◽  
High Concentration ◽  
Beneficial Role ◽  
Intimate Connection ◽  
Indri Indri ◽  
Lemur Species

AbstractHere, we investigated the possible linkages among geophagy, soil characteristics, and gut mycobiome of indri (Indri indri), an endangered lemur species able to survive only in wild conditions. The soil eaten by indri resulted in enriched secondary oxide-hydroxides and clays, together with a high concentration of specific essential micronutrients. This could partially explain the role of the soil in detoxification and as a nutrient supply. Besides, we found that soil subject to geophagy and indris’ faeces shared about 8.9% of the fungal OTUs. Also, several genera (e.g. Fusarium, Aspergillus and Penicillium) commonly associated with soil and plant material were found in both geophagic soil and indri samples. On the contrary, some taxa with pathogenic potentials, such as Cryptococcus, were only found in indri samples. Further, many saprotrophs and plant-associated fungal taxa were detected in the indri faeces. These fungal species may be involved in the digestion processes of leaves and could have a beneficial role in their health. In conclusion, we found an intimate connection between gut mycobiome and soil, highlighting, once again, the potential consequent impacts on the wider habitat.

scholarly journals POS0169 FETUIN-A AS A MARKER OF OSTEOPOROSIS AND OSTEOPOROTIC FRACTURES IN PATIENTS WITH RHEUMATOID ARTHRITIS

2021 ◽  
Vol 80 (Suppl 1) ◽  
pp. 297.2-297
Author(s):  
Y. Akhverdyan ◽  
E. Papichev ◽  
В. Zavodovsky ◽  
L. Seewordova ◽  
J. Polyakova
Keyword(s):  
Rheumatoid Arthritis ◽  
Blood Serum ◽  
Bone Mineralization ◽  
Expectant Management ◽  
Enzyme Linked Immunosorbent Assay ◽  
Healthy Individuals ◽  
High Concentration ◽  
Mineral Density ◽  
Fetuin A ◽  
Apparently Healthy

Background:The main mechanism of the effect of fetuin-A (FeA) on bone metabolism is its ability to bind calcium and proteins of the TGF-β family. It has been proven that the optimal concentration of TGF-β is necessary for the differentiation of bone tissue, and a high concentration inhibits bone mineralization. Thus, adequate osteogenesis is based on a complex balance between FeA and TGF-β levels. It can be assumed that the determination of the FeA level in the blood of patients with rheumatoid arthritis (RA) will help to optimize the diagnosis and predict the severity of osteoporosis (OP).Objectives:to study the possibility of predicting the development of osteoporosis and osteoporetic fractures in patients with RA, depending on the level of FeA in blood serum.Methods:We examined two groups of patients (52 patients with RA complicated by OP, 58 patients with RA without OP) and 30 apparently healthy individuals. The age of the surveyed ranged from 18 to 72 years, the average duration of the disease was 7.53±0.89 years. In both groups, the FeA level was determined by an indirect enzyme-linked immunosorbent assay using a commercial test. Bone mineral density (BMD) was also measured in both groups (Lunar DPX-NT GE).Results:The average FeA level in the group of RA patients was lower than in the group of conventionally healthy individuals (731.21±109.9 μg/ml and 812.9±76.2 μg/ml, respectively; F=13.34; p=0,0004). The normal FeA level was calculated using the formula M±2σ in the group of apparently healthy individuals and ranged from 653.55 μg/ml to 972.19 μg/ml.A decreased level of FeA was found in 20 patients (86.96%) in the group of patients with OP and only in 3 (13.04%) patients with RA who did not suffer from OP (p<0.001). It can be concluded that patients with RA and a low concentration of FeA in the blood serum have a higher risk of developing OP.In the group of patients with normal FeA level, osteoporetic fractures were observed in 12 (13.79%) patients and were absent in 75 (86.21%) patients (p<0.001). Thus, RA patients with normal serum FeA levels have a lower risk of osteoporetic fractures.We also found a positive significant correlation between the level of FeA and BMD in the femoral neck area. In the group of patients with a reduced FeA level (23 people), the mean BMD values were 0.732±0.022 g/cm2, and in the group of patients with a normal FeA level (87 patients) - 0.890±0.014 g/cm2 (p<0.001, F=27.663). The obtained values are in agreement with the literature data on the effect of the serum FeA concentration on the BMD values.Conclusion:We consider it expedient to determine the serum FeA concentration in patients with RA. At a FeA level of 653.55 μg/ml and below, a higher risk of developing OP and osteoporetic fractures can be predicted. In this case, the patient is shown a standard examination for osteoporosis. At values of 653.55 μg/ml and above, a more expectant management of the patient is allowed. Thus, by determining the serum concentration of FeA, it is possible to implement an integrated approach to the patient and to optimize the schemes for the diagnosis of OP in patients with RA.Disclosure of Interests:None declared

scholarly journals The Origin of Plasma-Derived Bacterial Extracellular Vesicles in Healthy Individuals and Patients with Inflammatory Bowel Disease: A Pilot Study

Genes ◽  
2021 ◽  
Vol 12 (10) ◽  
pp. 1636
Author(s):  
Emily Jones ◽  
Régis Stentz ◽  
Andrea Telatin ◽  
George M. Savva ◽  
Catherine Booth ◽  
...  
Keyword(s):  
Inflammatory Bowel Disease ◽  
Pilot Study ◽  
16S Rrna ◽  
Gene Sequencing ◽  
16S Rrna Gene Sequencing ◽  
Rrna Gene ◽  
Healthy Individuals ◽  
Inflammatory Bowel ◽  
Rrna Gene Sequencing ◽  
Blood Microbiota

The gastrointestinal tract harbors the gut microbiota, structural alterations of which (dysbiosis) are linked with an increase in gut permeability (“leaky gut”), enabling luminal antigens and bacterial products such as nanosized bacterial extracellular vesicles (BEVs) to access the circulatory system. Blood-derived BEVs contain various cargoes and may be useful biomarkers for diagnosis and monitoring of disease status and relapse in conditions such as inflammatory bowel disease (IBD). To progress this concept, we developed a rapid, cost-effective protocol to isolate BEV-associated DNA and used 16S rRNA gene sequencing to identify bacterial origins of the blood microbiome of healthy individuals and patients with Crohn’s disease and ulcerative colitis. The 16S rRNA gene sequencing successfully identified the origin of plasma-derived BEV DNA. The analysis showed that the blood microbiota richness, diversity, or composition in IBD, healthy control, and protocol control groups were not significantly distinct, highlighting the issue of ‘kit-ome’ contamination in low-biomass studies. Our pilot study provides the basis for undertaking larger studies to determine the potential use of blood microbiota profiling as a diagnostic aid in IBD.

Detection of α1 Integrin in Urine of Patients With Immunoglobulin A Nephropathy

2008 ◽  
Vol 56 (3) ◽  
pp. 581-586
Author(s):  
Jonathan Bank ◽  
Aharon Ben-David ◽  
Ram Doolman ◽  
Ben-Ami Sela ◽  
Ilan Bank
Keyword(s):  
Mesangial Cells ◽  
Kidney Diseases ◽  
Surface Membrane ◽  
Immunoglobulin A ◽  
Enzyme Linked Immunosorbent Assay ◽  
Dependent Manner ◽  
Healthy Individuals ◽  
Immunoglobulin A Nephropathy ◽  
A Cell ◽  
Dose Dependent

BackgroundThe α1β1 integrin is a cell surface membrane heterodimer composed of noncovalently linked α1 and β1 polypeptides that is up-regulated on activated and proliferating mesangial cells.MethodsA double-sandwich enzyme-linked immunosorbent assay that detects α1 integrin in a specific and dose-dependent manner at concentrations greater than 150 ng/mL was used to evaluate whether intact α1 polypeptides are secreted in the urine samples of 29 patients with various kidney diseases and in those of 5 healthy individuals.Resultsα1 Integrin was detected in 8 of the 29 patients including 3 of 3 patients with biopsy-proven immunoglobulin A nephropathy and 3 of 3 clinically suspected but non-biopsy-proven immunoglobulin A nephropathy with evidence of active nephritis. No α1 integrins were found in samples of 5 healthy controls.Conclusionsα1 Integrin polypeptides can be detected in human urine, particularly in immunoglobulin A nephropathy. Further extensive studies are required to clarify the significance of secretion of α1 integrins in urine of patients with kidney disease.

Chemical traits and antimicrobial activity of endemic Teucrium arduini L. from Mt Biokovo (Croatia)

2012 ◽  
Vol 7 (5) ◽  
pp. 941-947 ◽  
Author(s):  
Dario Kremer ◽  
Ivna Müller ◽  
Valerija Dunkić ◽  
Dubravka Vitali ◽  
Edith Stabentheiner ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Essential Oils ◽  
Bacterial Species ◽  
Glandular Trichomes ◽  
Toxic Metal ◽  
Antimicrobial Activities ◽  
Fungal Species ◽  
High Concentration ◽  
Sesquiterpene Hydrocarbons ◽  
Chemical Traits

AbstractChemical composition of the essential oil (analysed by GC-FID and GC-MS), the content of macroelements and trace elements (analysed by ICP-AES), and antimicrobial activities were investigated in Teucrium arduini L. from Mt Biokovo (Croatia). Additionally, a study on the types and distribution of glandular trichomes which produce essential oils was investigated. The oil was characterized by a high concentration of sesquiterpene hydrocarbons (68.5%) of which β-caryophyllene (32.9%) and germacrene D (16.4%) being the major compounds. Among the macroelements, the content of calcium was the highest (9772 mg/kg), while the content of sodium was the smallest (117.74 mg/kg). Among the micronutrients, the most represented element was iron (72.07 mg/kg). The content of each investigated toxic metal (As, Hg, Pb, Cd and Cr) was below permissible levels. The essential oils showed antimicrobial activity against bacterial species tested, with MIC values ranging from 6.25 mg/mL to 37.50 mg/mL. Fungal species were susceptible with MIC values from 7.81 mg/mL and 25.00 mg/mL.

scholarly journals Effects of contamination of blood specimens with liquid potassium-EDTA anticoagulant

2002 ◽  
Vol 39 (3) ◽  
pp. 273-280 ◽  
Author(s):  
D Fraser Davidson
Keyword(s):  
Best Practice ◽  
Biochemical Analysis ◽  
Binding Capacity ◽  
Blood Collection ◽  
High Concentration ◽  
Liquid Form ◽  
Calcium Magnesium ◽  
Blood Specimens ◽  
Iron Binding Capacity ◽  
Blood Collection Tubes

Background Use of the anticoagulant EDTA, in high-concentration liquid form, in blood collection tubes can lead to cross-contamination of routine biochemistry specimens. Methods In the present study, tests affected by EDTA in the sample were potassium, calcium, magnesium, unsaturated iron-binding capacity, bicarbonate, aspartate transaminase, alanine transaminase, lactate dehydrogenase, creatine kinase, alkaline phosphatase and amylase. The likely mechanisms are discussed. Conclusions In the interests of best practice, the identification of subtle contamination of blood specimens for biochemical analysis requires a sensitive method for measurement of EDTA itself.

scholarly journals Profiling copy number alterations in cell-free tumour DNA using a single-reference

2018 ◽  
Author(s):  
Alan J Robertson ◽  
Qinying Xu ◽  
Sarah Song ◽  
Devika Ganesamoorthy ◽  
Derek Benson ◽  
...  
Keyword(s):  
Cancer Patients ◽  
Copy Number ◽  
Read Depth ◽  
Reference Panel ◽  
Plasma Samples ◽  
Copy Number Profile ◽  
Copy Number Alterations ◽  
Healthy Individuals ◽  
A Cell ◽  
Sequencing Platforms

AbstractBackgroundThe accurate detection of copy number alterations from the analysis of circulating cell free tumour DNA (ctDNA) in blood is essential to realising the potential of liquid biopsies. However, currently available approaches require a large number of plasma samples from healthy individuals, sequenced using the same platform and protocols to act as a reference panel. Obtaining this reference panel can be challenging, prohibitively expensive and limits the ability to migrate to improved sequencing platforms and improved protocols.MethodsWe developed qCNV and sCNA-seq, two distinct tools that together provide a new approach for profiling somatic copy number alterations (sCNA) through the analysis of cell free DNA (cfDNA) without a reference panel. Our approach was designed to identify sCNA from cfDNA through the analysis of a single plasma sample and a matched normal DNA sample -both of which can be obtained from the same blood draw. qCNV is an efficient method for extracting read-depth from BAM files and sCNA-seq is a method that uses a probabilistic model of read depth to infer the copy number segmentation of the tumour. We compared the results from our pipeline to the established copy number profile of a cell-line, as well as the results from the plasma-Seq analysis of cfDNA-like mixtures and real, clinical data-sets.ResultsWith a single, unmatched, germline reference sample, our pipeline recapitulated the known copy number profile of a cell-line and demonstrated similar results to those obtained from plasma-Seq. With less than 1X genome coverage, our approach identified clinically relevant sCNA in samples with as little as 20 % tumour DNA. When applied to plasma samples from cancer patients, our pipeline identified clinically significant mutations.ConclusionsThese results show it is possible to identify therapeutically-relevant copy number mutations from plasma samples without the need to generate a reference panel from a large number of healthy individuals. Together with the range of sequencing platforms supported by our qCNV+sCNA-Seq pipeline, as well as the Galaxy implementation of this solution, this pipeline makes cfDNA profiling more accessible and makes it easier to identify sCNA from the plasma of cancer patients.

scholarly journals Effective removal of fly ash by Penicillium chrysogenum and determination of direct fly ash toxicity with Daphnia magna

2020 ◽  
Author(s):  
Burcu Ertit Taştan
Keyword(s):  
Fly Ash ◽  
Daphnia Magna ◽  
Penicillium Chrysogenum ◽  
Toxicity Assessment ◽  
Fungal Species ◽  
High Concentration ◽  
Element Analysis ◽  
Concentration Maximum ◽  
Effective Removal ◽  
Effect Level

Abstract This study demonstrates the removal of fly ash with Penicillium chrysogenum, a newly isolated species of fungus, and acute toxicity assessment with Daphnia magna. In the study, two different removal mechanisms were compared, both bio-removal and bio-sorption. Six different ash and three different biomass concentrations were used simultaneously. Although other fungal species in the literature failed at such a high concentration of fly ash, P. chrysogenum was able to tolerate it even at 10% concentration. The highest bio-removal yield was recorded as 100% at 0.5% fly ash concentration. Maximum bio-sorption yield was 95.27% after 24th hour. The evaluation results of fly ash bio-toxicity by D. magna showed that the no observed effect level (NOEL) was 0.2 mg/L and the low observed effect level (LOEL) was 0.5 mg/L. The element analysis, determined by ED-XRF, clarified that Ca, Si, Fe and S were the common elements in this ash. This is the first study in the literature where fly ash removal was carried out using P. chrysogenum for both bio-removal and bio-sorption and needs to be developed in the future.

scholarly journals Antinuclear Antibodies: Marker of Diagnosis and Evolution in Autoimmune Diseases

2018 ◽  
Vol 49 (3) ◽  
pp. e62-e73 ◽  
Author(s):  
Lucia M Sur ◽  
Emanuela Floca ◽  
Daniel G Sur ◽  
Marius C Colceriu ◽  
Gabriel Samasca ◽  
...  
Keyword(s):  
Autoimmune Disease ◽  
Autoimmune Diseases ◽  
Antinuclear Antibodies ◽  
Test Results ◽  
Correct Interpretation ◽  
Healthy Individuals ◽  
A Cell ◽  
Specific Antigens ◽  
Blood Specimens ◽  
Human Laryngeal Carcinoma

Abstract Antinuclear antibodies (ANAs) are autoantibodies that attack self-proteins within cell nucleus structures; their presence in serum may indicate an autoimmune disease. Also, positive ANA test results have been obtained in chronic infectious diseases, cancers, medication-related adverse events, and even healthy individuals. As a result, a correct interpretation of the presence of ANAs is needed. Identification of ANAs subtypes is an important part of clinical immunology. The presence of ANAs in patient blood specimens is detected using a cell-line substrate from human laryngeal carcinoma (HEp-2 cells). On this substrate, ANAs will bind specific antigens, which will lead to a suggestive fluorescent emission. The fluorescence patterns visualized under the fluorescence microscope can be correlated with certain subtypes of ANA and certain autoimmune diseases. Depending on the subtype of ANA present in the serum and the targeted antigen, several staining patterns are reported, namely, nuclear patterns, nucleolar patterns, cell cycle patterns, or cytoplasmatic patterns. Identification of a certain pattern can lead to diagnosis of a certain autoimmune disease.

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