P–408 Inadequate increase in Tim–3 on peripheral NK cells after blastocyst transfer is associated with early miscarriage

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
T Zhang ◽  
Y Zhao ◽  
C C Wing ◽  
X Chen ◽  
C C Wang ◽  
...  
Keyword(s):  
Immune Cells ◽  
Nkt Cells ◽  
University Teaching ◽  
Blastocyst Transfer ◽  
Ongoing Pregnancy ◽  
Time Points ◽  
Peripheral Lymphocytes ◽  
Significant Difference ◽  
Sustained Increase

Abstract Study question Whether the changing peripheral levels of Tim–3/Galectin–9 (Gal–9) and PD–1/PL–1 over 4 weeks after ET in ongoing pregnancies is different from pregnancies destined to miscarry. Summary answer A significant and sustained increase of Tim–3 in pNK cells was observed in pregnancies which were ongoing but not in pregnancies which later miscarried. What is known already The importance of maternal immune adaptation and tolerance to the implanting embryo, an allograft, has been extensively investigated for decades. Immune checkpoint molecules, like T-cell immunoglobulin mucin–3 (Tim–3) and programed cell death–1 (PD–1), are co-stimulatory receptors negatively regulating immune responses. During pregnancy, Tim–3 and PD–1 are expressed by several immune cells in the decidua and participate in the maternal-fetal immune interactions to mediate maternal immune tolerance through binding to their ligands Gal–9 and progressed death-ligand 1 (PD-L1) produced by trophoblast and immune cells. In addition to the implantation site, Tim–3 and PD–1 expressions in peripheral lymphocytes are modified during pregnancy. Study design, size, duration A prospective observational study includes 81 women who achieved ongoing pregnancy and 17 women who suffered from miscarriage after single day–5 blastocyst transfer. All the subjects were recruited from November 2018 to January 2020 in a university teaching hospital. Participants/materials, setting, methods Women undergoing single blastocyst transfer after in-vitro-fertilization (IVF) or intracytoplasmic sperm injection (ICSI) treatment were recruited on the day of ET following informed, written consent. They had serial blood samples taken on the day of ET, and 4, 5, 6 and 7 weeks of gestation for measurement of (1) membranous Tim–3 and PD–1 expression on various peripheral lymphocytes by flow cytometry; and (2) serum concentrations of ligands Gal–9 and PD-L1 by ELISA. Main results and the role of chance The comparisons between two groups showed there was no significant difference between the 2 groups in baseline levels among all the parameters measured. In women who achieved ongoing pregnancy, a significant and sustained increase of Tim–3 in either peripheral NK (pNK) subsets was observed at 4-week, 5-week, 6-week and 7-week gestations compared to the baseline (Tim–3+CD56dimNK 39.14±1.51%, 41.14±1.62%, 41.34±1.94%, and 41.69±2.12% vs. 30.27±1.49%; Tim–3+CD56brihgtNK cells, 24.54±1.71%, 25.43±1.54%, 27.26±1.88% and 24.70±1.64% vs. 19.08±1.13%), and the concentration of serum PD-L1 was significantly increased at 6-week and 7-week gestations (48.33±17.78 pg/ml, 52.53±20.60 pg/ml) when compared to the day of blastocyst transfer (41.40±16.01 pg/ml). The expressions of Tim–3 in T, NKT cells and PD–1 in NK, T, NKT cells were not significantly changed across the 5 time points. In women who conceived but later miscarried, all the parameters examined from 4–7 weeks of gestation were not significantly different when compared with the baseline measurement. The only measurement which showed a significant difference between the 2 groups and across all time points after ET was the proportion of Tim–3+CD56dimNK cells which was significantly higher in women who achieved ongoing pregnancies compared with women who destined to miscarry from 4 to 7 weeks of gestation. Limitations, reasons for caution It is uncertain if the observation would be different between miscarriage associated with aneuploid embryo or euploid embryo as we had not been able to obtain karyotyping result in most of the miscarriage cases. Wider implications of the findings: Our preliminary observation suggests that the proportion of Tim–3+pNK cells as early as 4-week gestation could be a potential immuno-bio-marker to predict if a pregnancy is likely to progress normally or result in a miscarriage. Clearly, the finding in this study needs to be confirmed in a larger cohort study Trial registration number not applicable.

scholarly journals Factor H (FH) Is Released Slowly and Continuously from Human Endothelial Cells (ECs): FH Is Not Packaged in Weibel-Palade Bodies or Secreted in Response to EC Stimulation

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4175-4175
Author(s):  
Sarah E Sartain ◽  
Nancy A Turner ◽  
Hui Shiu-Ki ◽  
Charles G. Minard ◽  
Joel L Moake
Keyword(s):  
Research Funding ◽  
Complement Components ◽  
Alexa Fluor ◽  
Time Points ◽  
Significant Difference ◽  
The Mean ◽  
Von Willebrand ◽  
Fund Research

Abstract Introduction Ultra-large von Willebrand factor (ULVWF) strings are synthesized in ECs, packaged in Weibel-Palade Bodies (WPBs), and secreted by stimulated ECs. Complement components studied to date [C3, factor (F) B, FD, FP, FH, FI, C5] are released slowly and continuously from human umbilical vein endothelial cell (HUVEC) cytoplasm and are not packaged in WPBs (PLoS One. 2013;8(3):e59372). In contrast, a recent report (Blood. 2014;123(1):121-5) contended that FH co-localizes with VWF in the WPBs. If this were so, it could have therapeutic importance for the treatment of atypical hemolytic uremic syndrome (aHUS) resulting from deficiency of FH because it might be possible to increase circulating FH levels transiently by administration of the WPB secretagogue, des-amino-D-arginine vasopressin (DDAVP). Hypothesis FH is not co-localized with VWF in WBPs, but rather is released slowly and continuously from EC cytoplasm regardless of cell stimulation. Methods Immunofluorescent Microscopy HUVECs were stimulated with histamine and stained with rabbit anti-VWF plus secondary donkey anti-rabbit Alexa Fluor IgG-488. The cells were then fixed and stained with goat-anti FH plus secondary chicken anti-goat Alexa Fluor IgG-647. The nuclei were detected with DAPI. In vitro VWF and FH from HUVECs HUVECs either were, or were not, stimulated with histamine. Supernatant was collected a variety of times over 7 hrs and assayed for VWF and FH antigen levels by ELISA. VWF assay antibodies (polyclonal): (1) capture, rabbit anti-human VWF (Ramco); (2) detection, goat anti-human VWF (Bethyl) and rabbit anti-goat IgG-HRP (Invitrogen). FH assay antibodies: (1) capture, polyclonal goat anti-human FH (Advanced Research Technologies); (2) detection, monoclonal mouse anti-human FH (Pierce, Thermo Scientific) and polyclonal goat anti-mouse IgG-HRP (Invitrogen). In vivo VWF and FH Plasma samples were obtained from 6 pediatric patients with von Willebrand disease (VWD) being tested for EC release of WPB VWF in response to DDAVP. For each patient, 1 sample was obtained prior to DDAVP administration, and 2 other samples were obtained 1 and 4 hours later. VWF levels were measured in each sample using standard clinical laboratory procedure at an affiliated hospital. FH antigen levels were quantified by ELISA, as above. Results Using non-overlapping spectral secondary detection antibody pairs, VWF was seen in clusters in HUVEC WPBs (Fig. 1A). In contrast, FH was distributed throughout the HUVEC cytoplasm (Fig. 1B). The VWF and FH images did not overlap, indicating that VWF and FH did not co-localize in the WPBs. Fig 1. Immunofluorescent images of HUVECs stained for VWF and FH. Fig 1. Immunofluorescent images of HUVECs stained for VWF and FH. Histamine addition to HUVECs in vitro resulted in ~ 4-fold increases in VWF secreted from HUVEC WPBs at 30 min and 1 hour, and 2-fold increases at 3 hours (Fig 2A). In contrast, FH release was slow and continuous, regardless of histamine stimulation, suggesting that FH is located in EC cytoplasm and is not stored in WPBs (Fig. 2B). Fig 2. In vitro VWF and FH release from ECs under non-stimulated and histamine-stimulated conditions. Fig 2. In vitro VWF and FH release from ECs under non-stimulated and histamine-stimulated conditions. In all 6 patient samples, VWF antigen increased significantly at 1-hour post-DDAVP administration (Fig. 3A). In contrast, FH antigen levels did not change significantly at hour 1 or hour 4, compared to hour 0, indicating that FH is not co-localized and secreted along with VWF from the WPBs of stimulated ECs in vivo (Fig. 3B). Fig 3. (A) Mean responses of VWF antigen to DDAVP, in vivo, with 95% CIs.The mean response was significantly greater 1-hour and 4-hours post-DDAVP compared with baseline (P=0.0085 and 0.0079, respectively). After 1-hour post-DDAVP, the mean response was 201% greater (95% CI: 129%, 314%) than baseline. After 4-hours, the mean response was 174% greater (95% CI: 123%, 247%) than baseline. (B) Responses of FH to DDAVP, in vivo. There was no statistically significant difference in FH response between time points (P=0.77). Fig 3. (A) Mean responses of VWF antigen to DDAVP, in vivo, with 95% CIs.The mean response was significantly greater 1-hour and 4-hours post-DDAVP compared with baseline (P=0.0085 and 0.0079, respectively). After 1-hour post-DDAVP, the mean response was 201% greater (95% CI: 129%, 314%) than baseline. After 4-hours, the mean response was 174% greater (95% CI: 123%, 247%) than baseline. (B) Responses of FH to DDAVP, in vivo. There was no statistically significant difference in FH response between time points (P=0.77). Conclusions We used immunofluorescent microscopy and ELISA assays on samples obtained in vitro and in vivo to demonstrate that FH is not packaged in, or secreted from, the WPBs of stimulated human ECs. FH is, therefore, similar to all other complement components studied to date in that it is released slowly and continuously from ECs and is not influenced by cell stimulation. DDAVP is unlikely to be a viable treatment option for patients with aHUS secondary to deficiency or inhibition of FH. Disclosures Sartain: Hemostasis and Thrombosis Research Society: Research Funding. Turner:Mary R Gibson Foundation: Research Funding; Hinkson Memorial Fund : Research Funding. Moake:Mary R Gibson Foundation: Research Funding; Hinkson Memorial Fund: Research Funding.

scholarly journals Biofilm-coated microbeads and the mouse ear skin: An innovative model for analysing anti-biofilm immune response in vivo

PLoS ONE ◽  
2020 ◽  
Vol 15 (12) ◽  
pp. e0243500
Author(s):  
Léo Sauvat ◽  
Aizat Iman Abdul Hamid ◽  
Christelle Blavignac ◽  
Jérôme Josse ◽  
Olivier Lesens ◽  
...  
Keyword(s):  
Real Time ◽  
Medical Devices ◽  
Immune Cells ◽  
Inflammatory Responses ◽  
Late Time ◽  
Average Cell ◽  
Time Points ◽  
Mouse Ear

Owing to its ability to form biofilms, Staphylococcus aureus is responsible for an increasing number of infections on implantable medical devices. The aim of this study was to develop a mouse model using microbeads coated with S. aureus biofilm to simulate such infections and to analyse the dynamics of anti-biofilm inflammatory responses by intravital imaging. Scanning electron microscopy and flow cytometry were used in vitro to study the ability of an mCherry fluorescent strain of S. aureus to coat silica microbeads. Biofilm-coated microbeads were then inoculated intradermally into the ear tissue of LysM-EGFP transgenic mice (EGFP fluorescent immune cells). General and specific real-time inflammatory responses were studied in ear tissue by confocal microscopy at early (4-6h) and late time points (after 24h) after injection. The displacement properties of immune cells were analysed. The responses were compared with those obtained in control mice injected with only microbeads. In vitro, our protocol was capable of generating reproducible inocula of biofilm-coated microbeads verified by labelling matrix components, observing biofilm ultrastructure and confirmed in vivo and in situ with a matrix specific fluorescent probe. In vivo, a major inflammatory response was observed in the mouse ear pinna at both time points. Real-time observations of cell recruitment at injection sites showed that immune cells had difficulty in accessing biofilm bacteria and highlighted areas of direct interaction. The average speed of cells was lower in infected mice compared to control mice and in tissue areas where direct contact between immune cells and bacteria was observed, the average cell velocity and linearity were decreased in comparison to cells in areas where no bacteria were visible. This model provides an innovative way to analyse specific immune responses against biofilm infections on medical devices. It paves the way for live evaluation of the effectiveness of immunomodulatory therapies combined with antibiotics.

scholarly journals comparison between Cleavage Stage versus Blastocyst Stage Embryo Transfer in an Egyptian Cohort Undergoing in vitro Fertilization: A possible Role for Laser Assisted Hatching

2011 ◽  
Vol 5 ◽  
pp. CMRH.S7735 ◽  
Author(s):  
Sherif F. Hendawy ◽  
TA Raafat
Keyword(s):  
Embryo Transfer ◽  
Intracytoplasmic Sperm Injection ◽  
Blastocyst Transfer ◽  
Assisted Hatching ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Implantation Rates ◽  
Group Ii ◽  
In Pregnancy

Background Extended in vitro embryo culture and blastocyst transfer have emerged as essential components of the advanced reproductive technology armamentarium, permitting selection of more advanced embryos considered best suited for transfer. Aim of study The aim of this study was to compare between cleavage stage and blastocyst stage embryo transfer in patients undergoing intracytoplasmic sperm injection, and to assess the role of assisted hatching technique in patients undergoing blastocyst transfer. Patients and methods This study was carried out on two groups. Group I: 110 patients who underwent 120 cycles of intracytoplasmic sperm injection with day 2-3 embryo transfer—for unexplained infertility or male factor within the previous 3 years. Their data obtained retrospectively from medical records. Group II: 46 age matched infertile female patients undergoing 51 intracytoplasmic sperm injection cycles for similar causes. Patients in Group II were further subdivided into 2 equal subgroups; Group Ila (23 patients), which had laser assisted hatching and Group IIb (23 patients), which did not have assisted hatching. All patients had an infertility workup including basal hormonal profile, pelvic ultrasound, hysterosalpingogram and/or laparoscope and semen analysis of the patient's partner. All patients underwent controlled ovarian hyperstimulation: Using long protocol of ovulation induction. Laser assisted hatching was done for blastocysts of 23 patients. Results Comparison between both groups as regards the reproductive outcome showed a significant difference in pregnancy and implantation rates, both being higher in group II ( P < 0.05) Comparison between both subgroups as regards the reproductive outcome showed a highly significant difference in pregnancy and implantation rates, both being higher in Group IIa ( P < 0.01). There was also a significantly higher rate of multiple pregnancies among Group IIa ( P < 0.05). Conclusion Blastocyst transfer is a successful and improved alternative for patients with multiple failed in vitro fertilization attempts, associated with a significant increase in pregnancy and implantation rates. Furthermore, laser assisted hatching increases implantation and clinical pregnancy rates.

scholarly journals Effect of evaporation-induced osmotic changes in culture media in a dry-type incubator on clinical outcomes in in vitro fertilization-embryo transfer cycles

2020 ◽  
Vol 47 (4) ◽  
pp. 284-292
Author(s):  
Hee-Jun Chi ◽  
Jun-Sang Park ◽  
Chang-Seok Yoo ◽  
Su-Jin Kwak ◽  
Ho-Jeong Son ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Clinical Outcomes ◽  
Embryo Transfer ◽  
Culture Media ◽  
Culture Conditions ◽  
Ongoing Pregnancy ◽  
Vitro Fertilization ◽  
Significant Difference ◽  
Ivf Et

Objective: This study investigated whether adding outer-well medium to inhibit osmotic changes in culture media in a dry-type incubator improved the clinical outcomes of in vitro fertilization-embryo transfer (IVF-ET) cycles. Methods: In culture dishes, the osmotic changes in media (20 µL)-covered oil with or without outer-well medium (humid or dry culture conditions, respectively) were compared after 3 days of incubation in a dry-type incubator. One-step (Origio) and G1/G2 (Vitrolife) media were used. Results: The osmotic changes in the dry culture condition (308 mOsm) were higher than in the humid culture conditions (285–290 mOsm) after 3 days of incubation. In day 3 IVF-ET cycles, although the pregnancy rate did not significantly differ between the dry (46.2%) and humid culture (52.2%) groups, the rates of abortion and ongoing pregnancy were significantly better in the humid culture group (2.3% and 50.2%, respectively) than in the dry culture group (8.3% and 37.8%, respectively, p<0.05). In day 5 IVF-ET cycles, the abortion rate was significantly lower in the humid culture group (2.2%) than in the dry culture group (25.0%, p<0.01), but no statistically significant difference was observed in the rates of clinical and ongoing pregnancy between the dry (50% and 25.0%, respectively) and humid culture groups (59.5% and 57.3%, respectively) because of the small number of cycles. Conclusion: Hyperosmotic changes in media occurred in a dry-type incubator by evaporation, although the medium was covered with oil. These osmotic changes were efficiently inhibited by supplementation of outer-well medium, which resulted in improved pregnancy outcomes.

scholarly journals Activation of Human Osteoblasts via Different Bovine Bone Substitute Materials With and Without Injectable Platelet Rich Fibrin in vitro

2021 ◽  
Vol 9 ◽  
Author(s):  
Solomiya Kyyak ◽  
Sebastian Blatt ◽  
Eik Schiegnitz ◽  
Diana Heimes ◽  
Henning Staedt ◽  
...  
Keyword(s):  
Metabolic Activity ◽  
Bone Substitute ◽  
Bone Morphogenetic Protein 2 ◽  
Bovine Bone ◽  
Human Osteoblasts ◽  
Time Points ◽  
Significant Difference ◽  
Bone Substitute Materials ◽  
Substitute Materials

IntroductionThe aim of the in vitro study was to compare the effect of four bovine bone substitute materials (XBSM) with and without injectable platelet-reach fibrin for viability and metabolic activity of human osteoblasts (HOB) as well as expression of alkaline phosphatase (ALP), bone morphogenetic protein 2 (BMP-2), and osteonectin (OCN).Materials and MethodsCerabone® (CB), Bio-Oss® (BO), Creos Xenogain® (CX) and MinerOss® X (MO) ± i-PRF were incubated with HOB. At day 3, 7, and 10, cell viability and metabolic activity as well as expression of ALP, OCN, and BMP-2, was examined.ResultsFor non-i-PRF groups, the highest values concerning viability were seen for CB at all time points. Pre-treatment with i-PRF increased viability in all groups with the highest values for CB-i-PRF after 3 and 7 and for CX-i-PRF after 10 days. For metabolic activity, the highest rate among non-i-PRF groups was seen for MO at day 3 and for CB at day 7 and 10. Here, i-PRF groups showed higher values than non-i-PRF groups (highest values: CB + i-PRF) at all time points. There was no difference in ALP-expression between groups. For OCN expression in non-i-PRF groups, CB showed the highest values after day 3, CX after day 7 and 10. Among i-PRF-groups, the highest values were seen for CX + i-PRF. At day 3, the highest BMP-2 expression was observed for CX. Here, for i-PRF groups, the highest increase was seen for CX + i-PRF at day 3. At day 7 and 10, there was no significant difference among groups.ConclusionXBSM sintered under high temperature showed increased HOB viability and metabolic activity through the whole period when compared to XBSM manufactured at lower temperatures. Overall, the combination of XBSM with i-PRF improved all cellular parameters, ALP and BMP-2 expression at earlier stages as well as OCN expression at later stages.

Differential response of factor VIII and protein C expression following multimodal therapy for esophageal carcinoma

2007 ◽  
Vol 25 (18_suppl) ◽  
pp. 15106-15106
Author(s):  
M. Byrne ◽  
B. M ◽  
J. O’ Donnell ◽  
B. White ◽  
J. Kennedy ◽  
...  
Keyword(s):  
Risk Factor ◽  
Factor Viii ◽  
Oesophageal Cancer ◽  
Protein C ◽  
Protein S ◽  
Endothelial Damage ◽  
Time Points ◽  
Increased Risk ◽  
Significant Difference

15106 Background: Tumour growth is associated with development of a hypercoaguable state and increased risk of thrombosis. Factor VIII plasma concentration has been shown to be an independent and dose-dependant risk factor for venous thromboembolism. Von Willebrand factor, the carrier molecule of circulating FVIII is expressed by vascular endothelial cells and in vitro observations indicate that chemotherapy induces endothelial damage with subsequent dysfunction. This study assessed FVIII, Protein C and Protein S response following multi modal therapy for oesophageal carcinoma. Methods: Fifty oesophageal cancer patients were studied, (multimodal n= 25, surgery only, n=25), at time points 0 (pre-operatively, following the first and final cycles of chemoradiation, and on days 1, 3, 7, 14, 21 and 28 days, and at 3 and 6 months postoperatively. Blood samples were collected and centrifuged at 3000g for 20 minutes to botain platelet poor plasmad. Factor VIII levels were measured by one stage clotting assay with factor VIII-deficient plasma. Automated functional assays for Protein C and Protein S in plasma were performed. Results: Factor FVIII levels were significantly elevated in the multimodal cohort following the 1st (p<0.008) and 2nd (p<0.001) cycles of chemoradiotherapy and at all time points following surgery, compared to baseline readings (p<0.001). Similar elevation was noted in the surgery only patients. No significant difference in factor FVIII levels between the multimodal group and surgery only patients was seen, except on day 21 (p < 0.04). Preliminary findings indicate a significant difference between the multimodal and surgery only cohorts in Protein C (0.74–1.32 IU/ml) levels (p < 0.05) at two timepoints. Conclusion: Factor VIII serum levels are significantly elevated following preoperative chemoradiotherapy. The pattern of response post oesophagectomy did not differ between the two cohorts. Elevated levels are a risk factor for both primary and recurrent venous thrombolism. The precise pathogenetic mechanism and cause of this elevation remains to be elucidated. Further studies investigating Protein C activity and, relating Factor VIII to thrombosis and inflammation in the oesophageal cancer patient are required. No significant financial relationships to disclose.

scholarly journals Mitochondrial DNA Content May Not Be a Reliable Screening Biomarker for Live Birth After Single Euploid Blastocyst Transfer

2021 ◽  
Vol 12 ◽  
Author(s):  
Xuanyou Zhou ◽  
Xueli Liu ◽  
Weihui Shi ◽  
Mujin Ye ◽  
Songchang Chen ◽  
...  
Keyword(s):  
Mitochondrial Dna ◽  
Live Birth ◽  
Embryo Implantation ◽  
Live Birth Rate ◽  
Blastocyst Stage ◽  
Blastocyst Transfer ◽  
Embryo Viability ◽  
Potential Biomarker ◽  
Significant Difference

An increasing number of studies have related the mitochondrial DNA (mtDNA) content to embryo viability and transfer outcomes. However, previous studies have focused more on the relationship between mtDNA and embryo implantation, few studies have studied the effect of the mtDNA content on live birth. In the study, we investigated whether mtDNA content is a reliable screening biomarker for live birth after single blastocyst transfer. A total of 233 couples with 316 blastocyst stage embryos undergoing in vitro fertilization treatment and pre-implantation genetic testing analysis were included in the study. All embryos were chromosomally normal and had undergone single-embryo transfers. There was no significant difference observed in the blastocyst mtDNA content among the live birth, miscarriage and non-implanted groups (p=0.999), and the mtDNA content in blastocysts from the miscarriage and live birth groups was similar [median (interquartile range), 1.00*108(7.59*107- 1.39*108) vs 1.01*108 (7.37*107- 1.32*108)]. Similarly, no significant association was observed between mtDNA content and embryo implantation potential (p=0.965). After adjusting for multiple confounders in a logistic regression analysis with generalized estimating equations, no associations between mtDNA content and live birth were observed in all blastocysts, Day-5 and Day-6 blastocysts (p=0.567, p=0.673, p=0.165, respectively). The live birth rate was not significantly different between blastocysts with an elevated mtDNA content and blastocysts with a normal mtDNA content (26.7% vs 33.6% p=0.780). Additionally, there was no linear correlation between the mtDNA content and maternal age (p=0.570). In conclusion, the mtDNA content does not seem to be a potential biomarker for embryo transfer outcomes (i.e., implantation and live birth) based on the existing testing tools. Embryos with an elevated mtDNA content also have development potential for successful live birth.

scholarly journals Assessment of Antiplaque and Anti-gingivitis Efficacy of Mouthwashes Prepared from Neem and Mango Extracts

2020 ◽  
Author(s):  
Nikita V Dandekar ◽  
J Jasmin Winnier
Keyword(s):  
Minimum Inhibitory Concentration ◽  
Streptococcus Mutans ◽  
Inhibitory Concentration ◽  
Plaque Index ◽  
Gingival Index ◽  
Time Points ◽  
Significant Difference ◽  
Salivary Ph

Objectives: This study aimed to determine the minimum inhibitory concentration (MIC) of mouthwashes prepared from neem and mango extracts against Streptococcus mutans (S. mutans) in vitro. Their taste acceptability and effects on plaque index (PI), gingival index (GI) and salivary pH were also evaluated and compared with chlorhexidine (CHX) in children. Materials and Methods: Dry extracts of neem and mango twigs were prepared and their MIC against S. mutans was determined. The effective MIC was used to prepare mouthwashes from the two extracts. Three parallel groups of children (n=30) used either neem, mango or 0.2% CHX mouthwash for 21 days. The PI and GI were recorded at baseline and at 7 and 21 days. The salivary pH and taste acceptability were also assessed. Results: The MIC of both extracts was achieved at 25% concentration. There was a significant difference between the GI score of mango group compared with neem and CHX at the three time points (P<0.001). There was no significant difference between the neem and mango groups in PI (P=0.674). There was no significant difference among the three groups in salivary pH either (P=0.817). Intragroup comparison showed significant reductions in PI, GI and salivary pH in all the three groups after 21 days (P<0.001). Conclusion: Neem and mango mouthwashes can be used as effective alternatives to CHX in children.

scholarly journals Bisphenol A Release from Orthodontic Clear Aligners: An In-Vitro Study

2021 ◽  
Vol 03 (03) ◽  
pp. 1-1
Author(s):  
Sydney Katras ◽  
◽  
Dejian Ma ◽  
Ayman al Dayeh ◽  
David Tipton ◽  
...  
Keyword(s):  
Bisphenol A ◽  
Artificial Saliva ◽  
In Vitro Study ◽  
Gastric Fluid ◽  
Duration Of Treatment ◽  
Synthetic Compound ◽  
Developmental Defects ◽  
Time Points ◽  
Significant Difference

Bisphenol A (BPA) is a widely used synthetic compound that has been identified as an endocrine disruptor. It has been linked to adverse health effects such as developmental defects, infertility in both men and women, cardiovascular disease, and obesity. There has been increased interest in BPA’s effects on the developing fetus and data has revealed that doses below the presumed safe dose can produce harmful effects. Orthodontic clear aligner therapy is a popular treatment modality that involves the patient wearing sets of plastic aligners up to 22 hours per day for the duration of treatment. The BPA release from these aligners was rarely investigated. The objective of this study was to detect, quantify, and compare the amount of BPA released by 3 popular brands of orthodontic aligners in artificial saliva, artificial gastric fluid, and ethanol. Equal amounts of SmileDirectClub®, Invisalign®, and Essix Ace® aligners were placed in sterile glass vials and submerged in 5.5mL artificial saliva, artificial gastric fluid, and ethanol. Samples were incubated at 37°C and 1 mL aliquots were removed at various time points. Samples were prepared and analyzed using high performance liquid chromatography tandem mass spectrometry. BPA was released from all three brands of aligners with great variability. Peak BPA concentrations were detected in artificial saliva; Smile Direct Club® produced 5.0 ng/mL after 20 days, Invisalign® released 3.5ng/mL after 24 hours, and Essix Ace® released 6.3ng/mL after 10 days of incubation in artificial saliva. There was no significant difference in BPA concentration between the 3 types of aligners in the 3 media. When comparing time points, there was a significant increase of BPA release in the first 24 hours after incubation compared to the baseline (p<0.001). There is potential BPA release from orthodontic aligners. There was no significant difference in the amount of BPA released between the three types of aligners at any time point. The majority of BPA release occurred during the first 24 hours.

scholarly journals ANXA1 as a Prognostic and Immune Microenvironmental Marker for Gliomas Based on Transcriptomic Analysis and Experimental Validation

2021 ◽  
Vol 9 ◽  
Author(s):  
Zhongxiao Lin ◽  
Min Wen ◽  
Enxing Yu ◽  
Xiao Lin ◽  
Hua Wang ◽  
...  
Keyword(s):  
Differentially Expressed Genes ◽  
Immune Cells ◽  
Cox Regression ◽  
Interaction Network ◽  
Prognostic Indicator ◽  
The Cancer Genome Atlas ◽  
Differentially Expressed ◽  
Cox Regression Analysis ◽  
Significant Difference

The tumor microenvironment (TME) plays an important role in the growth and invasion of glioma. This study aimed to analyze the composition of the immune microenvironment in glioma samples and analyze the important differentially expressed genes to identify novel immune-targeted therapy for glioma. We downloaded transcriptomic data of 669 glioma samples from The Cancer Genome Atlas database. CIBERSORT and ESTIMATE methods were used to calculate the proportion of tumor-infiltrating immune cells and ratio of immune and stromal components in the TME. The differentially expressed genes (DEGs) were screened by comparing the genes expressed by both stromal and immune cells. Annexin A1 (ANXA1) was determined to be an important prognostic indicator through the common overlap of univariate Cox regression analysis and protein–protein interaction network analysis. The proportion of tumor-infiltrating immune cells, calculated by CIBERSORT algorithm, had a significant difference in distribution among the high and low ANXA1 expression groups, indicating that ANXA1 could be an important immune marker of TME. Furthermore, ANXA1 level was positively correlated with the histopathological factors and negatively related to the survival of glioma patients based on the analysis of multiple databases. Finally, in vitro experiments verified that antagonizing ANXA1 expression promoted cell apoptosis and inhibited the invasion and migration capacities of glioma cells. Therefore, ANXA1 due to its immune-related functions, can be an important prognostic indicator and immune microenvironmental marker for gliomas. Further studies are warranted to confirm ANXA1 as a potential immunotherapeutic target for gliomas.

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