scholarly journals Improvement of Bacterial Vaginosis by Oral Lactobacillus Supplement: A Randomized, Double-Blinded Trial

2021 ◽  
Vol 11 (3) ◽  
pp. 902
Author(s):  
Ta-Chin Lin ◽  
I-Ling Hsu ◽  
Wan-Hua Tsai ◽  
Yi-Chih Chu ◽  
Lung-Ching Kuan ◽  
...  
Keyword(s):  
Bacterial Vaginosis ◽  
In Vitro Selection ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Beneficial Effects ◽  
Vaginal Microflora ◽  
Improvement Effect ◽  
Double Blinded ◽  
Blinded Trial

Bacterial vaginosis (BV) is the most common vaginal infection globally, with a high recurrent rate after antibiotic treatment. Probiotics consumption is known to improve BV with different efficacy among species or strains. After in vitro selection of Lactobacillus strains with growth inhibition and preventing adhesion to HeLa cervical epithelial cells, a randomized and double-blinded trial of two Lactobacillus formula, namely, VGA-1 and VGA-2, in BV patients with Nugent scores of 4–10 was conducted. Among 37 subjects who completed the trial, we observed significantly decreased Nugent scores in both VGA-1 (n = 18) and VGA-2 (n = 19) consumption groups. VGA-1 consumption significantly improved vaginal discharge odor/color and itching at both 2-week and 4-week-consumption, but those only observed after a 4-week-consumption in the VGA-2 group. We also observed a tendency to reduce recurrent rates among enrolled participants after VGA-1 or VGA-2 consumption. The improvement effect of VGA-1/VGA-2 was associated with the significant reduction of interleukin-6 expression after 4-week-consumption and the restoration of normal vaginal microflora by quantitative polymerase chain reaction analysis. In conclusion, VGA-1 or VGA-2 displayed beneficial effects in BV patients, but the VGA-1 formula showed a better efficacy, potentially used for BV intervention.

scholarly journals CircATRNL1 promotes epithelial–mesenchymal transition in endometriosis by upregulating Yes-associated protein 1 in vitro

2020 ◽  
Vol 11 (7) ◽  
Author(s):  
Dandan Wang ◽  
Yajuan Luo ◽  
Guangwei Wang ◽  
Qing Yang
Keyword(s):  
Epithelial Mesenchymal Transition ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
In Vitro Assays ◽  
Migration And Invasion ◽  
Circular Rnas ◽  
Mesenchymal Transition ◽  
Inhibitory Function ◽  
Ishikawa Cells

Abstract Endometriosis is a common and benign gynecological disorder but exhibits malignant features. However, the underlying pathogenesis and pathophysiology of endometriosis remain unclear. Circular RNAs have been demonstrated to participate in the occurrence and progression of multiple diseases. This study was aimed to explore the roles of circATRNL1 in endometriosis in vitro. Based on the results of reverse transcription-quantitative polymerase chain reaction analysis, we found significant upregulation of circATRNL1 and Yes-associated protein 1 (YAP1), while downregulation of miR-141-3p and miR-200a-3p in ectopic tissues compared to eutopic tissues. The immunohistochemistry and western blot analysis showed differentially expressed epithelial–mesenchymal transition (EMT) markers between EuEM and EcEM tissues. The in vitro assays indicated that overexpression of circATRNL1 could promote the proliferation, migration, and invasion of Ishikawa cells, and induce EMT process, while circATRNL1 silencing showed the opposite effect. The mechanical investigation indicated that circATRNL1 upregulated YAP1 by sponging miR-141-3p and miR-200a-3p. Gain-of-function assays validated the inhibitory function of miR-141-3p and miR-200a-3p in endometriosis. The results of rescue assays confirmed the function of circATRNL1–miR-141-3p/miR-200a-3p–YAP1 axis on Ishikawa cells. Our findings demonstrate that abnormal upregulation of circATRNL1 regulates cell proliferation and motility and promotes EMT process via the miR-141-3p/miR-200a-3p–YAP1 axis in vitro, which could contribute to the progression of endometriosis.

scholarly journals Effect of Nanostructured Scaffold on Human Adipose-Derived Stem Cells: Outcome of In Vitro Experiments

Nanomaterials ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1822 ◽  
Author(s):  
Marina Borgese ◽  
Ludovica Barone ◽  
Federica Rossi ◽  
Mario Raspanti ◽  
Roberto Papait ◽  
...  
Keyword(s):  
Stem Cells ◽  
Culture Medium ◽  
Quantitative Polymerase Chain Reaction ◽  
Enzyme Linked Immunosorbent Assay ◽  
Adipose Derived Stem Cells ◽  
In Vitro Experiments ◽  
Fatty Acid Binding ◽  
Beneficial Effects ◽  
Angiogenesis Process

This work is addressed to provide, by in vitro experiments, results on the repercussion that a nanostructured scaffold could have on viability, differentiation and secretion of bioactive factors of human adipose-derived stem cells (hASCs) when used in association to promote angiogenesis, a crucial condition to favour tissue regeneration. To achieve this aim, we evaluated cell viability and morphology by MTT (3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide) assay and microscopy analysis, respectively. We also investigated the expression of some of those genes involved in angiogenesis and differentiation processes utilizing quantitative polymerase chain reaction (qPCR), whereas the amounts of Vascular Endothelial Growth Factor A, Interleukin 6 and Fatty Acid-Binding Protein 4 secreted in the culture medium, were quantified by enzyme-linked immunosorbent assay (ELISA). Results suggested that, in the presence of the scaffold, cell proliferation and the exocytosis of factors involved in the angiogenesis process are reduced; by contrast, the expression of those genes involved in hASC differentiation appeared enhanced. To guarantee cell survival, the construct dimensions are, generally, smaller than clinically required. Furthermore, being the paracrine event the primary mechanism exerting the beneficial effects on injured tissues, the use of conditioned culture medium instead of cells may be convenient.

Tannin-rich forage as a methane mitigation strategy for cattle and the implications for rumen microbiota

2021 ◽  
Vol 61 (1) ◽  
pp. 26 ◽  
Author(s):  
Gisele M. Fagundes ◽  
Gabriela Benetel ◽  
Mateus M. Carriero ◽  
Ricardo L. M. Sousa ◽  
James P. Muir ◽  
...  
Keyword(s):  
Production Systems ◽  
Condensed Tannin ◽  
Quantitative Polymerase Chain Reaction ◽  
Methanogenic Archaea ◽  
Polymerase Chain Reaction Analysis ◽  
Gliricidia Sepium ◽  
Desmodium Ovalifolium ◽  
Ruminal Bacteria ◽  
Rumen Microbiota

Context Methane from ruminant livestock systems contributes to the greenhouse effect on the environment, which justifies the adoption of novel feed strategies that mitigate enteric emissions. Aims We investigated the effects of the condensed tannin (CT)-rich legumes Flemingia macrophylla, Leucaena leucocephala, Stylosanthes guianensis, Gliricidia sepium, Cratylia argentea, Cajanus cajan, Desmodium ovalifolium, Macrotyloma axillare, Desmodium paniculatum and Lespedeza procumbens on in vitro methane emissions and rumen microbiota for beef cattle. Methods Four rumen-cannulated Nellore cattle grazing a tropical grass pasture were used as inoculum donors. Key results Real-time quantitative polymerase chain reaction analysis revealed that the abundance of Ruminococcus flavefaciens, methanogenic archaea and protozoa populations were reduced (P £ 0.05), whereas total ruminal bacteria were enhanced in the presence of CT. Our study also revealed a positive (P £ 0.05) relationship between CT and Fibrobacter succinogenes abundance. Reactive CT from L. leucocephala, D. paniculatum and L. procumbens resulted in decreased (P £ 0.05) isoacid content and methane production. Conclusions L. leucocephala, D. paniculatum and L. procumbens have the potential to suppress rumen methanogenesis. However, in vitro fermentation of L. leucocephala resulted in greater (P £ 0.05) degradability percentages than the other two species. Implications CT in legume species will have potential as part of an overall nutritional strategy to manipulate rumen microbiota and mitigate enteric methanogenesis in livestock production systems.

scholarly journals The Viability and Anti-Inflammatory Effects of Hyaluronic Acid-Chitlac-Tracimolone Acetonide-β-Cyclodextrin Complex on Human Chondrocytes

Cartilage ◽  
2020 ◽  
pp. 194760352090865 ◽  
Author(s):  
Elena Tarricone ◽  
Rossella Elia ◽  
Elena Mattiuzzo ◽  
Alessia Faggian ◽  
Assunta Pozzuoli ◽  
...  
Keyword(s):  
Gene Expression ◽  
Hyaluronic Acid ◽  
Articular Chondrocytes ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Human Chondrocytes ◽  
Cyclodextrin Complex ◽  
Modified Chitosan ◽  
Anti Inflammatory

Objective To compare the effects of the complex triamcinolone acetonide-hydroxypropyl-β-cyclodextrin (TA-CD) on in vitro inflamed primary human articular chondrocytes in the presence or absence of the mixture hyaluronic acid-Chitlac, a lactose-modified chitosan (HA-CTL). Design Changes in cell viability and pro-inflammatory cytokines gene expression were analyzed in human chondrocytes using an in vitro model of macrophage-mediated inflammation. Human monocytes U937 were differentiated to macrophages by phorbol 12-myristate 13-acetate (PMA) and lipopolysaccharides (LPS). The anti-inflammatory effects of the complex TA-CD and HA-CTL mixture were assessed on chondrocytes exposed for 24 hours to U937 conditioned medium (CM), by quantitative polymerase chain reaction analysis. Results The TA-CD viability was enhanced by the presence of the HA-CTL mixture in chondrocyte cultures. The exposure of cells to CM significantly increased interleukin-1β and interleukin-6 gene expression, and when the complex TA-CD was added to the inflamed cells, gene transcription of cytokines was restored to near baseline values, both in the presence or in the absence of HA-CTL mixture. Conclusion The addition of HA-CTL mixture significantly attenuated cytotoxicity induced by TA and preserved the anti-inflammatory effects, thus confirming the chondroprotective role of the HA-CTL mixture.

scholarly journals In Vivo Association of Ku with Mammalian Origins of DNA Replication

2001 ◽  
Vol 12 (11) ◽  
pp. 3386-3401 ◽  
Author(s):  
Olivia Novac ◽  
Diamanto Matheos ◽  
Felipe D. Araujo ◽  
Gerald B. Price ◽  
Maria Zannis-Hadjopoulos
Keyword(s):  
Dna Replication ◽  
Quantitative Polymerase Chain Reaction ◽  
Polymerase Chain Reaction Analysis ◽  
Origin Firing ◽  
Origin Binding Protein ◽  
M Phase ◽  
Polymerase Chain ◽  
In Cells

Ku is a heterodimeric (Ku70/86-kDa) nuclear protein with known functions in DNA repair, V(D)J recombination, and DNA replication. Here, the in vivo association of Ku with mammalian origins of DNA replication was analyzed by studying its association withors8 and ors12, as assayed by formaldehyde cross-linking, followed by immunoprecipitation and quantitative polymerase chain reaction analysis. The association of Ku with ors8 and ors12 was also analyzed as a function of the cell cycle. This association was found to be approximately fivefold higher in cells synchronized at the G1/S border, in comparison with cells at G0, and it decreased by approximately twofold upon entry of the cells into S phase, and to near background levels in cells at G2/M phase. In addition, in vitro DNA replication experiments were performed with the use of extracts from Ku80+/+ and Ku80−/− mouse embryonic fibroblasts. A decrease of ∼70% in in vitro DNA replication was observed when the Ku80−/− extracts were used, compared with the Ku80+/+ extracts. The results indicate a novel function for Ku as an origin binding-protein, which acts at the initiation step of DNA replication and dissociates after origin firing.

scholarly journals A novel microscopic method for analyzing gram-stained vaginal smears in the diagnosis of disorders of vaginal microflora

2015 ◽  
Vol 72 (8) ◽  
pp. 670-676 ◽  
Author(s):  
Dane Nenadic ◽  
Milos Pavlovic ◽  
Tatjana Motrenko
Keyword(s):  
Bacterial Vaginosis ◽  
Quantitative Polymerase Chain Reaction ◽  
Great Majority ◽  
Concordance Correlation ◽  
Vaginal Smears ◽  
Microscopic Method ◽  
Vaginal Microflora ◽  
Simple Tool ◽  
Pmn Leukocytes ◽  
Polymerase Chain

Background/Aim. The Nugent?s score is still the gold standard in the great majority of studies dealing with the assessment of vaginal flora and the diagnosis of bacterial vaginosis (BV). The aim of this study was to show that the analysis of Gram-stained vaginal samples under microscope at the magnification of ?200 (a novel microscopic method - NMM), as a fast and simple tool, easily applicable in everyday practice, better reflects complexity of vaginal microflora than the Nugent?s methodology (?1000). Methods. Gramstained vaginal smears from 394 asymptomatic pregnant women (24-28 week of pregnancy) were classified according to the Nugent?s microscopic criteria (immersion, magnification ?1000). The smears were then reexamined under immersion but at magnification ?200. All samples were classified into 6 groups according to semiquanititative assessment of numbers (cellularity) and the ratio of rod (length < 1.5 ?m) and small bacterial (< 1.5 ?m) forms: hypercellular (normal full - NF), moderately cellular (normal mid - NM), hypocellular (normal empty - NE), bacterial vaginosis full (BVF), bacterial vaginosis mid (BVM), and bacterial vaginosis empty (BVE). Also yeasts, coccae, bifido and lepto bacterial forms as well polymorphonuclear (PMN) leukocytes were identified. Results. According to the Nugent?s scoring, BV was found in 78, intermediate findings in 63, and yeasts in 48 patients. By our criteria BV was confirmed in 88 patients (37 BVF, 24 BVM, and 27 BVN). Generally, both tools proved to be highly concordant for the diagnosis of BV (Lin?s concordance correlation coefficient = 0.9852). In 40% of the women mixed flora was found: yeasts in 126 (32%), coccae in 145 (37%), bifido forms in 32 (8%) and lepto forms in 20 (5%). Almost a half of BV patients had also yeasts (39/88). Elevated PMN numbers were found in 102 (33%) patients with normal and in 36 (41%) women with BV. Conclusion. The newly described methodology is simpler to apply and much better reflects diversity of vaginal microflora. In this way it may be more valuable to molecular biologists and their attempts based on quantitative polymerase chain reaction (PCR) to define formulas for molecular diagnosis of bacterial vaginosis.

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