scholarly journals An In Vitro Approach to Studying the Microbial Community and Impact of Pre and Probiotics under Anorexia Nervosa Related Dietary Restrictions

Nutrients ◽  
2021 ◽  
Vol 13 (12) ◽  
pp. 4447
Author(s):  
Litai Liu ◽  
Carlos Poveda ◽  
Paul E. Jenkins ◽  
Gemma E. Walton
Keyword(s):  
Anorexia Nervosa ◽  
Microbial Community ◽  
In Situ Hybridisation ◽  
Well Being ◽  
Fluorescence In Situ Hybridisation ◽  
Dietary Restrictions ◽  
Gastrointestinal Problems ◽  
The Impact ◽  
Nutritional Changes

Individuals with anorexia nervosa (AN) often suffer psychological and gastrointestinal problems consistent with a dysregulated gut microbial community. Psychobiotics have been postulated to modify microbiota and improve mental well-being and gut symptoms, but there is currently a lack of evidence for such approaches in AN. The aim of this study was to use an in vitro colonic model to evaluate the impact of dietary restrictions associated with AN on the intestinal ecosystem and to assess the impact of pre and probiotic intervention. Bacteriology was quantified using flow cytometry combined with fluorescence in situ hybridisation and metabolic end products (including neurotransmitters) by gas chromatography and liquid chromatography mass spectrometry Consistent with previous research, the nutritional changes significantly reduced total microbiota and metabolites compared with healthy conditions. Pre and probiotic supplementation on restricted conditions enhanced the microbial community and modulated metabolic activity to resemble that of a healthy diet. The model system indicates that nutritional changes associated with AN can impact the microbial community, and that these changes can, at least in part, be restored through the use of pre and probiotic interventions.

Short-term effects of carbon source on the competition of polyphosphate accumulating organisms and glycogen accumulating organisms

2004 ◽  
Vol 50 (10) ◽  
pp. 139-144 ◽  
Author(s):  
A. Oehmen ◽  
Z. Yuan ◽  
L.L. Blackall ◽  
J. Keller
Keyword(s):  
Carbon Source ◽  
In Situ Hybridisation ◽  
Operating Conditions ◽  
Fluorescence In Situ Hybridisation ◽  
Biological Phosphorus Removal ◽  
Short Term ◽  
Uptake Rates ◽  
Polyphosphate Accumulating Organisms ◽  
The Impact ◽  
Short Term Effects

The effectiveness of enhanced biological phosphorus removal (EBPR) systems is directly affected by the competition of polyphosphate accumulating organisms (PAOs) and glycogen accumulating organisms (GAOs). This study investigated the short-term effects of carbon source on PAO and GAO performance. The tests were designed to clearly determine the impact of volatile fatty acid (VFA) composition on the performance of two types of biomass, one enriched for PAOs and the other for GAOs. The two populations were enriched in separate reactors using identical operating conditions and very similar influent compositions with acetate as the sole carbon source. The only difference was that a very low level of phosphorus was present in the influent to the GAO reactor. The abundance of PAOs and GAOs was quantified using fluorescence in-situ hybridisation. The results clearly show that there are some very distinctive differences between PAOs and GAOs in their ability to utilise different carbon substrates. While both are able to take up acetate rapidly and completely, the GAOs are far slower at consuming propionate than the PAOs during short-term substrate changes. This provides a potentially highly valuable avenue to influence the competition between PAOs and GAOs. Other VFAs studied seem to be less usable in the short term by both PAOs and GAOs, as indicated by their much lower uptake rates.

scholarly journals The Impact of Low-Level Iron Supplements on the Faecal Microbiota of Irritable Bowel Syndrome and Healthy Donors Using In Vitro Batch Cultures

Nutrients ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 3819
Author(s):  
Carlos Poveda ◽  
Dora I. A. Pereira ◽  
Marie C. Lewis ◽  
Gemma E. Walton
Keyword(s):  
Irritable Bowel Syndrome ◽  
Gut Microbiota ◽  
Iron Supplementation ◽  
Ferrous Sulphate ◽  
Fluorescence In Situ Hybridisation ◽  
Batch Cultures ◽  
Low Level ◽  
Irritable Bowel ◽  
The Impact

Ferrous iron supplementation has been reported to adversely alter the gut microbiota in infants. To date, the impact of iron on the adult microbiota is limited, particularly at low supplementary concentrations. The aim of this research was to explore the impact of low-level iron supplementation on the gut microbiota of healthy and Irritable Bowel Syndrome (IBS) volunteers. Anaerobic, pH-controlled in vitro batch cultures were inoculated with faeces from healthy or IBS donors along with iron (ferrous sulphate, nanoparticulate iron and pea ferritin (50 μmol−1 iron)). The microbiota were explored by fluorescence in situ hybridisation coupled with flow cytometry. Furthermore, metabolite production was assessed by gas chromatography. IBS volunteers had different starting microbial profiles to healthy controls. The sources of iron did not negatively impact the microbial population, with results of pea ferritin supplementation being similar to nanoparticulate iron, whilst ferrous sulphate led to enhanced Bacteroides spp. The metabolite data suggested no shift to potentially negative proteolysis. The results indicate that low doses of iron from the three sources were not detrimental to the gut microbiota. This is the first time that pea ferritin fermentation has been tested and indicates that low dose supplementation of iron is unlikely to be detrimental to the gut microbiota.

scholarly journals CircSERPINE2 protects against osteoarthritis by targeting miR-1271 and ETS-related gene

2019 ◽  
Vol 78 (6) ◽  
pp. 826-836 ◽  
Author(s):  
Shuying Shen ◽  
Yizheng Wu ◽  
Junxin Chen ◽  
Ziang Xie ◽  
Kangmao Huang ◽  
...  
Keyword(s):  
In Situ Hybridisation ◽  
Rabbit Model ◽  
Pathological Process ◽  
Luciferase Reporter ◽  
Circular Rnas ◽  
Fluorescence In Situ Hybridisation ◽  
Related Gene ◽  
Downstream Target

ObjectivesCircular RNAs (circRNA) expression aberration has been identified in various human diseases. In this study, we investigated whether circRNAs could act as competing endogenous RNAs to regulate the pathological process of osteoarthritis (OA).MethodsCircRNA deep sequencing was performed to the expression of circRNAs between OA and control cartilage tissues. The regulatory and functional role of CircSERPINE2 upregulation was examined in OA and was validated in vitro and in vivo, downstream target of CircSERPINE2 was explored. RNA pull down, a luciferase reporter assay, biotin-coupled microRNA capture and fluorescence in situ hybridisation were used to evaluate the interaction between CircSERPINE2 and miR-1271-5 p, as well as the target mRNA, E26 transformation-specific-related gene (ERG). The role and mechanism of CircSERPINE2 in OA was also explored in rabbit models.ResultsThe decreased expression of CircSERPINE2 in the OA cartilage tissues was directly associated with excessive apoptosis and imbalance between anabolic and catabolic factors of extracellular matrix (ECM). Mechanistically, CircSERPINE2 acted as a sponge of miR-1271-5 p and functioned in human chondrocytes (HCs) through targeting miR-1271-5 p and ERG. Intra-articular injection of adeno-associated virus-CircSERPINE2-wt alleviated OA in the rabbit model.ConclusionsOur results reveal an important role for a novel circRNA-CircSERPINE2 in OA progression. CircSERPINE2 overexpression could alleviate HCs apoptosis and promote anabolism of ECM through miR-1271-ERG pathway. It provides a potentially effective therapeutic strategy for OA progression.

scholarly journals In vitro evaluation of the impact of human background microbiota on the response to Bifidobacterium strains and fructo-oligosaccharides

2013 ◽  
Vol 110 (11) ◽  
pp. 2030-2036 ◽  
Author(s):  
Silvia Arboleya ◽  
Nuria Salazar ◽  
Gonzalo Solís ◽  
Nuria Fernández ◽  
Miguel Gueimonde ◽  
...  
Keyword(s):  
Human Population ◽  
Well Being ◽  
Full Term ◽  
Population Groups ◽  
Culture Model ◽  
Faecal Samples ◽  
Predominant Factor ◽  
Microbial Groups ◽  
The Impact

The microbial colonisation of the infant gut begins immediately after birth and is essential for the development of the intestine, the immune system and later well-being. Important differences have been reported in the characteristics of such microbiota in different infant population groups. In the present study, we employed an in vitro faecal batch culture model using faeces from different human population groups (adults and full-term breast-fed, full-term formula-fed and preterm infants) to determine the influence that the addition of four bifidobacterial strains and fructo-oligosaccharides (FOS) exerts on the profile of SCFA measured by GC as well as on the levels of some relevant intestinal microbial groups by quantitative PCR during incubation. Differences were found in the levels of SCFA and intestinal microbial groups in the faecal cultures depending on the human group origin of the faecal samples (P< 0·05), this being a predominant factor, compared with bifidobacteria or FOS added, in determining microbiota dynamics. These results exhibit the importance of the initial characteristics of the basal intestinal microbiota in the effect exerted by bifidobacteria or FOS that are added and highlight the need to design probiotics targeting specific human population groups.

The composition and metabolism of faecal microbiota is specifically modulated by different dietary polysaccharides and mucin: an isothermal microcalorimetry study

2018 ◽  
Vol 9 (1) ◽  
pp. 21-34 ◽  
Author(s):  
K. Adamberg ◽  
K. Kolk ◽  
M. Jaagura ◽  
R. Vilu ◽  
S. Adamberg
Keyword(s):  
Illumina Miseq ◽  
Well Being ◽  
Isothermal Microcalorimetry ◽  
Faecal Microbiota ◽  
Bifidobacterium Adolescentis ◽  
Functional Changes ◽  
Dietary Fibres ◽  
Faecal Samples ◽  
The Impact

The metabolic activity of colon microbiota is specifically affected by fibres with various monomer compositions, degree of polymerisation and branching. The supply of a variety of dietary fibres assures the diversity of gut microbial communities considered important for the well-being of the host. The aim of this study was to compare the impact of different oligo- and polysaccharides (galacto- and fructooligosaccharides, resistant starch, levan, inulin, arabinogalactan, xylan, pectin and chitin), and a glycoprotein mucin on the growth and metabolism of faecal microbiota in vitro by using isothermal microcalorimetry (IMC). Faecal samples from healthy donors were incubated in a phosphate-buffered defined medium with or without supplementation of a single substrate. The generation of heat was followed on-line, microbiota composition (V3-V4 region of the 16S rRNA using Illumina MiSeq v2) and concentrations of metabolites (HPLC) were determined at the end of growth. The multiauxic power-time curves obtained were substrate-specific. More than 70% of all substrates except chitin were fermented by faecal microbiota with total heat generation of up to 8 J/ml. The final metabolite patterns were in accordance with the microbiota changes. For arabinogalactan, xylan and levan, the fibre-affected distribution of bacterial taxa showed clear similarities (e.g. increase of Bacteroides ovatus and decrease of Bifidobacterium adolescentis). The formation of propionic acid, an important colon metabolite, was enhanced by arabinogalactan, xylan and mucin but not by galacto- and fructooligosaccharides or inulin. Mucin fermentation resulted in acetate, propionate and butyrate production in ratios previously observed for faecal samples, indicating that mucins may serve as major substrates for colon microbial population. IMC combined with analytical methods was shown to be an effective method for screening the impact of specific dietary fibres on functional changes in faecal microbiota.

scholarly journals Understanding the Blogging Practices of Women Undergoing In Vitro Fertilization (IVF): A Discourse Analysis of Women’s IVF Blogs

2017 ◽  
Author(s):  
Elizabeth Orr ◽  
Susan Jack ◽  
Wendy Sword ◽  
Sandra Ireland ◽  
Lois Ostolosky
Keyword(s):  
Discourse Analysis ◽  
In Vitro Fertilization ◽  
Positive Impact ◽  
Emotional Health ◽  
Well Being ◽  
Fertility Treatment ◽  
Vitro Fertilization ◽  
The Rich ◽  
The Impact

Infertility and its associated treatments, including in vitro fertilization (IVF), can have a profound impact on the emotional health and well-being of women desiring to become mothers. Researchers have measured the impact of infertility and described the experience of infertility and its treatment, leaving the rich descriptions of the IVF experience as captured in women’s blogs to be explored. This discourse analysis describes the blogging practices of women undergoing IVF, exploring both the content and function of the IVF blog discourse. Data were collected from the text of seven women’s blogs (n=1,149 blog posts) and resulted in four main functions of the discourse: creation of and connection to a community, emotional support, blogging as therapy, and creation of an IVF resource. Findings suggest that blogging can have a positive impact on the psychosocial consequences experienced by women in fertility treatment.

Liquid biopsies to track trastuzumab resistance in metastatic HER2-positive gastric cancer

Gut ◽  
2018 ◽  
Vol 68 (7) ◽  
pp. 1152-1161 ◽  
Author(s):  
De-Shen Wang ◽  
Ze-Xian Liu ◽  
Yun-Xin Lu ◽  
Hua Bao ◽  
Xue Wu ◽  
...  
Keyword(s):  
Gastric Cancer ◽  
In Situ Hybridisation ◽  
Targeted Sequencing ◽  
In Vivo Studies ◽  
Fluorescence In Situ Hybridisation ◽  
Trastuzumab Resistance ◽  
Liquid Biopsies ◽  
Her2 Positive

ObjectiveTo monitor trastuzumab resistance and determine the underlying mechanisms for the limited response rate and rapid emergence of resistance of HER2+ metastatic gastric cancer (mGC).DesignTargeted sequencing of 416 clinically relevant genes was performed in 78 paired plasma and tissue biopsy samples to determine plasma-tissue concordance. Then, we performed longitudinal analyses of 97 serial plasma samples collected from 24 patients who were HER2+  to track the resistance during trastuzumab treatment and validated the identified candidate resistance genes.ResultsThe results from targeted sequencing-based detection of somatic copy number alterations (SCNA) of HER2 gene were highly consistent with fluorescence in situ hybridisation data, and the detected HER2 SCNA was better than plasma carcinoembryonic antigen levels at predicting tumour shrinkage and progression. Furthermore, most patients with innate trastuzumab resistance presented high HER2 SCNA during progression compared with baseline, while HER2 SCNA decreased in patients with acquired resistance. PIK3CA mutations were significantly enriched in patients with innate resistance, and ERBB2/4 genes were the most mutated genes, accounting for trastuzumab resistance in six (35.3%) and five (29.4%) patients in baseline and progression plasma, respectively. Patients with PIK3CA/R1/C3 or ERBB2/4 mutations in the baseline plasma had significantly worse progression-free survival. Additionally, mutations in NF1 contributed to trastuzumab resistance, which was further confirmed through in vitro and in vivo studies, while combined HER2 and MEK/ERK blockade overcame trastuzumab resistance.ConclusionLongitudinal circulating tumour DNA sequencing provides novel insights into gene alterations underlying trastuzumab resistance in HER2+mGC.

scholarly journals Novel lncRNA UPLA1 mediates tumorigenesis and prognosis in lung adenocarcinoma

2020 ◽  
Vol 11 (11) ◽  
Author(s):  
Xiaoyang Han ◽  
Hua Jiang ◽  
Jianni Qi ◽  
Jiamei Li ◽  
Jinghan Yang ◽  
...  
Keyword(s):  
Lung Adenocarcinoma ◽  
In Situ Hybridisation ◽  
Vital Role ◽  
Luciferase Reporter ◽  
Fluorescence In Situ Hybridisation ◽  
Biological Functions ◽  
Pulldown Assay

AbstractWith the development of molecular biotechnology and sequencing techniques, long non-coding RNAs (lncRNAs) have been shown to play a vital role in a variety of cancers including lung cancer. In our previous study, we used RNA sequencing and high-content screening proliferation screening data to identify lncRNAs that were significantly associated with tumour biological functions such as LINC01426. Herein, based on previous work, we report a novel lncRNA UPLA1 (upregulation promoting LUAD-associated transcript-1), which has not been explored or reported in any previous studies. Our results showed that UPLA1 is highly expressed and regulates important biological functions in lung adenocarcinoma. In vitro experiments revealed that UPLA1 promoted the migration, invasion, and proliferation abilities, and is related to cell cycle arrest, in lung adenocarcinoma cells. Moreover, the upregulation of UPLA1 significantly improved the growth of tumours in vivo. We identified that UPLA1 was mainly located in the nucleus using fluorescence in situ hybridisation, and that it promoted Wnt/β-catenin signalling by binding to desmoplakin using RNA pulldown assay and mass spectrometry. Additionally, luciferase reporter assay revealed that YY1 is the transcription factor of UPLA1 and suppressed the expression of UPLA1 as a transcriptional inhibitor. This finding provides important evidence regarding the two roles of YY1 in cancer. Furthermore, in situ hybridisation assay results showed that UPLA1 was closely related to the prognosis and tumour, node, metastasis (TNM) stage of lung adenocarcinoma. In summary, our results suggest that the novel lncRNA UPLA1 promotes the progression of lung adenocarcinoma and may be used as a prognostic marker, and thus, has considerable clinical significance.

scholarly journals Adipose tissue derived bacteria are associated with inflammation in obesity and type 2 diabetes

Gut ◽  
2020 ◽  
Vol 69 (10) ◽  
pp. 1796-1806 ◽  
Author(s):  
Lucas Massier ◽  
Rima Chakaroun ◽  
Shirin Tabei ◽  
Alyce Crane ◽  
Konrad David Didt ◽  
...  
Keyword(s):  
Type 2 Diabetes ◽  
Adipose Tissue ◽  
Immune Cell ◽  
In Situ Hybridisation ◽  
Rrna Gene ◽  
Fluorescence In Situ Hybridisation ◽  
Bacterial Dna ◽  
Tissue Specific

ObjectiveBacterial translocation to various organs including human adipose tissue (AT) due to increased intestinal permeability remains poorly understood. We hypothesised that: (1) bacterial presence is highly tissue specific and (2) related in composition and quantity to immune inflammatory and metabolic burden.DesignWe quantified and sequenced the bacterial 16S rRNA gene in blood and AT samples (omental, mesenteric and subcutaneous) of 75 subjects with obesity with or without type 2 diabetes (T2D) and used catalysed reporter deposition (CARD) – fluorescence in situ hybridisation (FISH) to detect bacteria in AT.ResultsUnder stringent experimental and bioinformatic control for contaminants, bacterial DNA was detected in blood and omental, subcutaneous and mesenteric AT samples in the range of 0.1 to 5 pg/µg DNA isolate. Moreover, CARD-FISH allowed the detection of living, AT-borne bacteria. Proteobacteria and Firmicutes were the predominant phyla, and bacterial quantity was associated with immune cell infiltration, inflammatory and metabolic parameters in a tissue-specific manner. Bacterial composition differed between subjects with and without T2D and was associated with related clinical measures, including systemic and tissues-specific inflammatory markers. Finally, treatment of adipocytes with bacterial DNA in vitro stimulated the expression of TNFA and IL6.ConclusionsOur study provides contaminant aware evidence for the presence of bacteria and bacterial DNA in several ATs in obesity and T2D and suggests an important role of bacteria in initiating and sustaining local AT subclinical inflammation and therefore impacting metabolic sequelae of obesity.

A Cytogenetic Approach to the Study of Genotoxic Effects of Fungicides: An in Vitro Study in Lymphocyte Cultures with Thiophanate-methyl

1996 ◽  
Vol 24 (4) ◽  
pp. 597-601
Author(s):  
Patrizia Hrelia ◽  
Carmela Fimognari ◽  
Fernanda Vigagni ◽  
Francesca Maffei ◽  
Giorgio Cantelli-Forti
Keyword(s):  
Human Peripheral Blood ◽  
Human Lymphocytes ◽  
In Situ Hybridisation ◽  
In Vitro Study ◽  
Peripheral Blood Lymphocytes ◽  
Fluorescence In Situ Hybridisation ◽  
Thiophanate Methyl ◽  
Mechanisms Of Toxicity ◽  
Genetic Level

This study was designed to evaluate the mechanisms of genetic damage by fungicides in cultured human peripheral blood lymphocytes by means of a molecular cytogenetic approach. For example, thiophanate-methyl (30μg/ml-300μg/ml) was shown to significantly induce chromosome aberrations and micronuclei in human lymphocytes cultured in vitro. Fluorescence in situ hybridisation with centromeric DNA probes demonstrated that most micronuclei induced by thiophanate-methyl did not show any centromeric signals, indicating a relatively stronger clastogenic activity. Results obtained with thiophanate-methyl showed that a comprehensive examination of the mechanisms of toxicity at the genetic level provides valuable information, which is of importance in the safety assessment of the fungicide.

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