Postimplantation Period Research Articles

Chondrogenesis of Mesenchymal Stem Cells through Local Release of TGF-β3 from Heparinized Collagen Biofabric.

Osteoarthritic degeneration of cartilage is a major social health problem. Tissue engineering of cartilage using combinations of scaffold and mesenchymal stem cells (MSCs) is emerging as an alternative to existing treatment options such as microfracture, mosaicplasty, allograft, autologous chondrocyte implantation, or total joint replacement. Induction of chondrogenesis in high-density pellets of MSCs is generally attained by soluble exogenous TGF-β3 in culture media, which requires lengthy in vitro culture period during which pellets gain mechanical robustness. On the other hand, a growth factor delivering and a mechanically robust scaffold material that can accommodate chondroid pellets would enable rapid deployment of pellets after seeding. Delivery of the growth factor from the scaffold locally would drive the induction of chondrogenic differentiation in the postimplantation period. Therefore, we sought to develop a biomaterial formulation that will induce chondrogenesis in situ, and compared its performance to soluble delivery in vitro. In this vein, a heparin-conjugated mechanically robust collagen fabric was developed for sustained delivery of TGF-β3. The amount of conjugated heparin was varied to enhance the amount of TGF-β3 uptake and release from the scaffold. The results showed that the scaffold delivered TGF-β3 for up to 8 days of culture, which resulted in 15-fold increase in GAG production, and six-fold increase in collagen synthesis with respect to the No TGF-β3 group. The resulting matrix was cartilage like, in that type II collagen and aggrecan were positive in the spheroids. Enhanced chondrogenesis under in situ TGF-β3 administration resulted in a Young's modulus of ∼600 kPa. In most metrics, there were no significant differences between the soluble delivery group and in situ heparin-mediated delivery group. In conclusion, heparin-conjugated collagen scaffold developed in this study guides chondrogenic differentiation of hMSCs in a mechanically competent tissue construct, which showed potential to be used for cartilage tissue regeneration. Impact statement The most significant finding of this study was that sustained release of TGF-β3 from heparinized collagen scaffold had chondroinductive effect on pelleted human mesenchymal stem cells (hMSCs). The effect was comparable to that observed in hMSC pellets that were cultured in chondrogenic media supplemented with TGF-β3. The stiffness of scaffolds at the baseline was about 50% that of native cartilage and over 28 days the combined stiffness of pellet/scaffold complex converged to the stiffness of native cartilage. These data indicate that the scaffold system can generate a load-bearing cartilage-like tissue by using hMSCs pellets in a mechanically competent framework.

Diets enriched in PUFAs at an early postimplantation stage prevent embryo resorptions and impaired mTOR signaling in the decidua from diabetic rats

Maternal diabetes increases the risk of embryo resorptions and impairs embryo development. Decidualization is crucial for embryo development and regulated by mTOR signaling. However, little is known about how maternal diabetes affects the decidua at early postimplantation stages and whether dietary treatments enriched in polyunsaturated fatty acids (PUFAs) can prevent decidual alterations. Here, we determined resorption rates, decidual mTOR pathways and markers of decidual function and remodeling in diabetic rats fed or not with diets enriched in PUFAs exclusively during the early postimplantation period. Pregestational streptozotocin-induced diabetic Albino Wistar rats and controls were fed or not with diets enriched in 6% sunflower oil or 6% chia oil (enriched in n-6 or n-3 PUFAs, respectively) on days 7, 8 and 9 of pregnancy and evaluated on day 9 of pregnancy. Maternal diabetes induced an 11-fold increase in embryo resorptions, which was prevented by both PUFAs-enriched diets despite no changes in maternal glycemia. The activity of mTOR pathway was decreased in the decidua from diabetic rats, an alteration prevented by the PUFAs-enriched diets. PUFAs-enriched diets prevented increased expression of Foxo1 (a negative regulator of mTOR) and reduced expression of miR-21 (a negative regulator of Foxo1). These diets also prevented reduced markers of decidual function (leukemia inhibitory factor and IGFBP1 expression and MMPs activity) in diabetic rat decidua. We identified the early post implantation as a crucial stage for pregnancy success, in which dietary PUFAs can protect diabetic pregnancies from embryo resorptions, decidual mTOR signaling impairments, and altered markers of decidual function and remodeling.

Optimal Bridging Strategy Post-Ventricular Assist Device Implantation Remains Unclear.

To the Editor: It was with enthusiasm that we read the article by Bates et al. describing bivalirudin utilization in a heterogeneous ventricular assist device (VAD) population.1 We were concerned, however, regarding their argument for the potential of bivalirudin as an initial bridging agent post-device implantation. No patients in their cohort received bivalirudin in this setting without being transitioned from either warfarin or heparin for either suspected or confirmed thrombosis or heparin-induced thrombocytopenia. Furthermore, the median time from VAD insertion to initiation of bivalirudin in their study was 116 days. Although we recognize the potential benefits of direct thrombin inhibitor therapy in the scenarios described within their cohort, the evidence for use of bivalirudin during the immediate postimplantation period as a bridge to oral anticoagulation is not well-elucidated. We have published the largest series to date of bivalirudin bridged patients (35) to oral anticoagulation after VAD placement.2 In our analysis, three different strategies were compared: bivalirudin, heparin, and no bridging immediately after VAD implantation. Our results suggested that bivalirudin was no different compared with heparin or no anticoagulation in terms of both thrombotic complications and bleeding events within 30 days of VAD implantation when controlling for international normalized ratio (INR) goals. In addition, the cost of heparin therapy was more than 40 times less than bivalirudin, although with the generic introduction of bivalirudin since the publication of our research, this number has certainly diminished. Overall, neither the Bates et al. data nor ours is sufficient to appropriately answer the question of which strategy is optimal to utilize post-VAD implantation.1,2 In addition, as the authors suggest, device type may also influence the benefits or weaknesses of various anticoagulant therapies. With the dramatic reduction in the rate of suspected of confirmed pump thrombosis observed in MOMENTUM-3, it is unclear if benefits observed in patients with older generation devices will translate to benefits in patients with the HeartMate 3.3 A large randomized trial, although welcome, is unlikely to be undertaken leaving a gap in the literature, which has yet to be filled for over a decade. Statements, however, on the utility of an approach without conclusive data, such as the use of bivalirudin in the post-implant period, and with some evidence showing that it has no discernable benefit should be made cautiously, if at all.

Prognostic Value of Cardiopulmonary Exercise Test Parameters in Ventricular Assist Device Therapy

<h3>Purpose</h3> Cardiopulmonary exercise test parameters are among the best prognosticators for medically managed advanced heart failure. However, the prognostic value of pre & post-implantation CPET parameters in LVAD therapy is unclear and is evaluated in this study. <h3>Methods</h3> Adult patients who were implanted with an LVAD and underwent CPET during the pre and/or post-implantation period were included to the study. Five parameters were derived from each CPET: vO₂max, oxygen uptake efficiency slope (OUES), vE/vCO₂min, vE/vCO₂max and vE/vCO₂ Slope. The independent predictive relationship between CPET parameters and clinical outcomes was evaluated with multivariate analysis. <h3>Results</h3> Pre & post-implantation CPET cohorts included 191 and 122 patients respectively (Table 1). Among pre-implantation CPET parameters: vO₂max was an independent predictor of in-hospital mortality; vO₂max and OUES were independent predictors of 1-year, 3-year and 5-year mortality, whereas vE/vCO₂min and vE/vCO₂ Slope were independent predictors of 5-year mortality (p<.05). No CPET parameter was associated with ICU or hospital length of stay, 30-day readmission or 90-day mortality (p>.05). Among post-implantation CPET parameters: vO₂max was an independent predictor of 5-year mortality, while vE/vCO₂min and vE/vCO₂max were independent predictors of 3-year mortality (p<.05) (Table 2). <h3>Conclusion</h3> CPET parameters derived from pre and post-implantation exercise tests have high predictive value for long-term outcomes in LVAD therapy. Except vO₂max, CPET parameters' predictive capacity for early post-implantation outcomes appears to be limited.

Xeno-free cryopreservation of adherent retinal pigmented epithelium yields viable and functional cells in vitro and in vivo

Age-related macular degeneration (AMD) is the primary cause of blindness in adults over 60 years of age, and clinical trials are currently assessing the therapeutic potential of retinal pigmented epithelial (RPE) cell monolayers on implantable scaffolds to treat this disease. However, challenges related to the culture, long-term storage, and long-distance transport of such implants currently limit the widespread use of adherent RPE cells as therapeutics. Here we report a xeno-free protocol to cryopreserve a confluent monolayer of clinical-grade, human embryonic stem cell-derived RPE cells on a parylene scaffold (REPS) that yields viable, polarized, and functional RPE cells post-thaw. Thawed cells exhibit ≥ 95% viability, have morphology, pigmentation, and gene expression characteristic of mature RPE cells, and secrete the neuroprotective protein, pigment epithelium-derived factor (PEDF). Stability under liquid nitrogen (LN2) storage has been confirmed through one year. REPS were administered immediately post-thaw into the subretinal space of a mammalian model, the Royal College of Surgeons (RCS)/nude rat. Implanted REPS were assessed at 30, 60, and 90 days post-implantation, and thawed cells demonstrate survival as an intact monolayer on the parylene scaffold. Furthermore, immunoreactivity for the maturation marker, RPE65, significantly increased over the post-implantation period in vivo, and cells demonstrated functional attributes similar to non-cryopreserved controls. The capacity to cryopreserve adherent cellular therapeutics permits extended storage and stable transport to surgical sites, enabling broad distribution for the treatment of prevalent diseases such as AMD.

Mid-Term Outcomes Following Percutaneous Pulmonary Valve Implantation Using the "Folded Melody Valve" Technique.

Background:The folded valve is a manual shortening of the Melody device, which has been validated as a valuable therapeutic option for the management of dysfunctional right ventricular outflow tracts needing a short valved stent. In this article, we aimed to evaluate, in a multicenter cohort, the mid-term outcomes of patients in whom a percutaneous pulmonary valve implantation was performed using the folded valve technique.Methods:A 2012 to 2018 retrospective multicenter study was performed in 7 European institutions. All patients who benefit from percutaneous pulmonary valve implantation with a folded Melody valve were included.Results:A total of 49 patients (median age, 19 years [range 4–56], 63% male) were included. The primary percutaneous pulmonary valve implantation indication was right ventricular outflow tract stenosis (n=19; 39%), patched native right ventricular outflow tracts were the most common substrate (n=15; 31%). The folded technique was mostly used in short right ventricular outflow tracts (n=28; 57%). Procedural success was 100%. After a median follow-up of 28 months (range, 4–80), folded Melody valve function was comparable to the immediate postimplantation period (mean transvalvular peak velocity=2.6±0.6 versus 2.4±0.6 m/s, P>0.1; only 2 patients had mild pulmonary regurgitation). Incidence rate of valve-related reinterventions was 2.1% per person per year (95% CI, 0.1%–3.9%). The probability of survival without valve-related reinterventions at 36 months was 90% (95% CI, 76%–100%).Conclusions:The folded Melody valve is a safe technique with favorable mid-term outcomes up to 6.5 years after implantation, comparable with the usual Melody valve implantation procedure. Complications and reinterventions rates were low, making this technique relevant in selected patients.

Leaf ethanolic extract of Etlingera hemesphaerica Blume mitigates defects in fetal anatomy and endochondral ossification due to mercuric chloride during the post-implantation period in Mus musculus.

This study aimed to investigate the effectiveness of leaf ethanolic extract of Etlingera hemisphaerica (LE3H) in reducing defects in fetal anatomy and endochondral ossification in mice induced by HgCl2 during the post-implantation period. Pregnant mice were divided into four groups, each consisting of 10 dams, and received drink and food ad libitum. The first group was administered LE3H (E1), the second one HgCl2 (E2), the third one HgCl2+LE3H (E3), and the fourth was control (E0), administered double-distilled water only. HgCl2 (5 mg/kg bw) was administrated by injection intraperitoneally on gestation day (GD)9 and LE3H (0.39 mg/g bw) was administered by gavage on GD10. The treated and control animals were killed by cervical dislocation on GD18, dissected, and the morphologically normal living fetuses (MNLF) were collected. The MNLF of E0, E1, E2, and E3 from 5 dams were fixed with Bouin solution, and observed using the free hand razor blade technique for soft tissue examination. The remaining MNLF were fixed with 96% ethanol, and then stained with Alizarin Red S and Alcian Blue for ossification examination. Index of length of ossified part (ILOP) of humerus, index of width of ossified part (IWOP) of humerus, ILOP of femur, and IWOP of femur were calculated. E2 had higher cases of anatomical defects (74,6%) than E3 (48.9%), E1 (15.0%), and E0 (0%). E2 had humerus IWOP of 0.82±0.03, which was significantly lower than that of E0 (0.89±0.04) and E1 (0.89±0.03), while that of E1 and E0 was not significantly different from each other. Meanwhile, IWOP in E3 (0.88±0.03) was significantly higher than that in E2, but not different from that in E1 and E0. Thus, LE3H mitigated defects in fetal anatomy and endochondral ossification induced by HgCl2 in mice.

Video head impulse test saccades and loss of cervical vestibular evoked myogenic potentials are late vestibular footprints of cochlear implantation.

Cochlear implantation may be complicated by concurrent injury to the vestibular apparatus, potentially resulting in disabling vertigo and balance problems. Information on vestibular function before implantation as measured by the video head impulse test (VHIT) and cervical vestibular evoked myogenic potentials (cVEMPs) is scarce and literature on long-term effects is non-existing. We aimed to evaluate how vestibular function was affected by cochlear implantation (CI), as measured by VHIT and cVEMPs in the late phase after implantation. Retrospective repeated measurement study. Among the 436 patients elected for CI surgery during 2013 to 2018, 45 patients met the inclusion criteria (CI recipients with a vestibular assessment prior to the first CI and a repeated vestibular assessment after the CI operation). VHIT and cVEMPs before and after cochlear implantation. Main outcome measures were vestibular function as evaluated by VHIT gain, saccades and cVEMPs. The mean time between first and second vestibular screening was 19 months. The mean VHIT gain on implanted ears was 0.79 before the operation and showed no change at follow-up (p = 0.65). Likewise, the number of abnormal VHIT gain values was equal before and after the operation (p = 0.31). Preoperatively, saccades were present on 12 ears (14%) compared with 25 ears (29%) postoperatively (p = 0.013) and were associated with significantly lower VHIT gain values. Preoperatively, positive cVEMPs were found in 20 CI ears (49%) and 24 contralateral ears (62%). 10 CI ears lost cVEMP postoperatively compared with 2 ears on the contralateral side (p = 0.0047). Even though VHIT gain has been reported to be affected in the immediate post-implantation period, the findings in this study show that VHIT gain is normalized in the long-term. However, cochlear implantation is associated with the occurrence of VHIT saccades in the long-term and these are associated with lower VHIT gain values. In addition, cVEMP responses are significantly reduced long-term on implanted ears, which agrees with other studies with shorter follow-up.

Intraoperative finding of immobile leaflet(s) following freshly implanted bioprosthetic valves: clinical characteristics and impact on outcomes

Abstract Funding Acknowledgements Type of funding sources: None. BACKGROUND Detection of immobile leaflets immediately following bioprosthetic valve implantation is a rare but important intraoperative finding. Restriction of leaflet movement can occur in the closed or open position, leading to abnormal prosthesis function. We sought to determine the clinical implications of immobile leaflets seen on intraoperative echocardiography. METHODS Patients with immobile leaflets identified on intra-operative/procedure echocardiography immediately post implantation between 2009-2020 were identified from an institutional database. All echocardiograms were reviewed de-novo to confirm immobile leaflets in the immediate post-implantation period. Identified cases were matched 1:2 to controls for age; sex; prosthesis position, model and size; and implantation approach (surgical vs. transcatheter). Nominal logistic regression and proportional hazards were used to analyze outcomes. RESULTS Thirty patients with immobile leaflets immediately post-bioprosthesis implantation were included. Clinical characteristics are summarized in the Table. Immobile leaflets were documented in procedural reports in only 18 (60%) patients. Moderate stenosis was present intraoperatively in 1 patient, none demonstrated ≥moderate regurgitation, and none resulted in immediate corrective action. In 3 (10%), valve re-intervention was required within 30 days due to symptomatic prosthesis dysfunction. Presence of restricted leaflet motion was associated with higher need for post-operative extracorporeal membrane oxygenation use (odds-ratio 7.3, p = 0.02) and composite end-point of death, valve re-replacement, prosthesis thrombosis, or cardiac hospitalizations (risk ratio 2.1, p = 0.03, Figure). CONCLUSION Immobile leaflet(s) immediately post-bioprosthetic valve implantation is an uncommon, under-reported, and under-treated phenomenon. Even in the absence of significant prosthetic valve dysfunction, it can be associated with worse post-operative course as well as worse outcomes. Baseline characteristics Age 76 (67-84) Sex, male 10 (33%) Surgical approach 25 (83%) Aortic 5 (17%) Mitral 12 (40%) Tricuspid 12 (40%) Pulmonary 1 (3%) Re-intervention within 10 days 3 (10%) Numbers are presented as median (interquartile range) or number (percentage). Abstract Figure. Clinical outcome of stuck leaflets

Inferior Vena Cava Filter Retrieval Trends: A Single-Center Experience.

Recognition of the adverse events of inferior vena cava filters (VCFs) has prompted the Food and Drug Administration (FDA) to issue safety warnings (2010 and 2014), advocating for removal, once the risk of pulmonary embolism has abated. Despite an initial increase in retrieval rates, these remain low (25–30% at 1 year in 2014). We retrospectively investigated retrieval trends in adults with VCFs placed between 2015 and 2018 at a single institution. The rate of retrievable VCF removal accounting for the competing risk of death was the main outcome. There were 494 VCFs placed (305 retrievable). The cumulative incidence of retrieval remained low (21% at 1 year), even after the second FDA warning (2014). Patients who resumed anticoagulation (AC) at any time were more likely to have retrieval (hazard ratio [HR] = 3.6, p < 0.01) and had higher retrieval rates at every time point (31.4 vs. 7.6% at 1 year). Advanced age (HR = 0.98 per year, p = 0.004), stroke (HR = 0.28, p = 0.028), and active malignancy (HR = 0.42, p = 0.006) predicted nonretrieval. Device-related complications were infrequent (<1%) but thrombotic complications occurred early and were more common for nonretrieved VCFs (17 vs. 12%, p = 0.29). Revision of guidelines to recommend active surveillance for the ability to tolerate AC in the immediate postimplantation period may improve retrieval rates.

Carbon nanohorn coating by electrodeposition accelerate bone formation on titanium implant

Direct contact between bone and implant materials is required for dental implants. Titanium is used for the implant material owing to its mechanical and biological properties. The anodisation as the surface treatment was employed to enhance osteogenesis around titanium. Moreover, carbon nanohorn (CNH), a type of nanometer-sized carbon material, was reported to promote the bone formation. Thus, it is expected that if the surface of anodised Ti (AnTi) is modified with CNHs, Ti-bone contact would be enhanced. In this study, the Ti surface was modified with CNHs by electrophoresis and obtained anodised titanium coated with CNHs (CNH/AnTi). In vitro, CNH/AnTi attracted osteoblastic cells more than AnTi, thereby the proliferation of osteoblastic cell was enhanced by CNH/AnTi more than by AnTi. In vivo, at 7 and 28 days after implantation of CNH/AnTi or AnTi into the rat femur, more aggressive bone formation was observed on the surface of CNH/AnTi than on AnTi. More importantly, the area where newly formed bone tissue directly attached to CNH/AnTi was significantly larger than that for AnTi, suggesting that “contact osteogenesis” was accelerated on CNH/AnTi during the early post-implantation period. CNH/AnTi would be advantageous especially for the early stages of bone regeneration after surgery.

P7 La adición de ácidos grasos poliinsaturados en el período de postimplantación temprana previene reabsorciones embrionarias y anomalías deciduales en ratas gestantes con diabetes

Introducción: la diabetes pregestacional incrementa el riesgo de reabsorciones embrionarias y afecta el desarrollo embrionario, de impacto en la programación fetal. La función decidual es crucial en el desarrollo postimplantatorio temprano y sus posibles anomalías son poco conocidas en la diabetes materna. Previamente evidenciamos que la vía de señalización de mTOR en la decidua es relevante para el crecimiento y desarrollo embrionario, y que interactúa con la vía de los PPAR, cuyos agonistas son nutrientes de naturaleza lipídica como los ácidos grasos poliinsaturados (PUFAs).Objetivos: determinar el índice de reabsorciones y evaluar la regulación de la vía de mTOR y marcadores de la función, y remodelación de la decidua en ratas con diabetes pregestacional que recibieron o no dietas enriquecidas en PUFAs exclusivamente en el período de postimplantación temprana.Materiales y métodos: ratas sanas o con diabetes (inducidas preconcepcionalmente con estreptozotocina, 55 mg/kg) fueron apareadas con machos sanos. En los días 7,8 y 9 de preñez recibieron una dieta enriquecida o no en 6% aceite de girasol o de chía (enriquecidas en PUFAs n-6 o n-3, respectivamente). En el día 9 de preñez se estudió el índice de reabsorción embrionaria y se explantó la decidua para estudiar la vía de mTOR, la expresión de su regulador FoxO1 y del microRNA-21 (miR-21) que regula a FoxO1, como así también marcadores de función y remodelación decidual (expresión de LIF y IGFBP1 y actividad gelatinasa de metaloproteasas (MMPs).

Longitudinal Changes in Health-related Quality of Life After 125I Low-dose-rate Brachytherapy for Localized Prostate Cancer.

The factors associated with longitudinal changes in health-related quality of life (HRQOL) are unclear. In this study we aimed to evaluate the longitudinal changes and predictors of HRQOL after 125I low-dose-rate brachytherapy (LDB) for localised prostate cancer (PCA). We evaluated 180 patients with localised PCA treated with LDB. The HRQOL was evaluated at 3 weeks before LDB and at 1, 3, 6, 12, 18, 24, 36, and 48 months after LDB using the International Prostate Symptom Score, Medical Outcome Study 8-Items Short Form Health Survey (SF-8), and University of California Los Angeles Prostate Cancer Index (UCLA-PCI). All HRQOL scores, except for UCLA-PCI sexual function and SF-8 mental component summary (MCS), were improved to baseline after an early transient deterioration. In contrast, the sexual function did not return to baseline after early deterioration. Meanwhile, the MCS scores showed no significant decline after implantation and trended upward. The prostate V100 and baseline UCLA-PCI sexual function scores predicted a clinically significant decrease in sexual function in the late post-implantation period. Most aspects of the HRQOL of PCA patients who underwent LDB improved to baseline. The results that V100 and baseline sexual function were predictors of late post-LDB may provide more accurate information for patients with preserved sexual function before treatment and for their partners.

CardioMEMS Device Implantation Reduces Repeat Hospitalizations In A Veterans Affairs Patient Population: A Single Center Experience

Introduction Little is known about the use of a wireless pulmonary artery pressure monitoring device, CardioMEMS, in the Veteran's Affair (VA) patient population. We report our results of the effectiveness of this wireless device in reducing heart failure re-admissions in the largest known VA patient cohort. Methods We retrospectively analyzed all patients with heart failure-related admissions pre- and post-implantation of their CardioMEMS device. Results A total of 63 patients (93% male, 44% Caucasian and 54% HFrEF), with NYHA Class 3 HF were followed for 12 months. At device implantation, average pulmonary artery systolic/ diastolic /mean pressures were 51/24/34 mgHg while at 6 months were 45/22/31 mmHg and 12 months were 43/21/29 mmHg. The total number of heart failure hospital admissions for patients who were followed for 12 months prior to device implantation was 130, while the total number for the post-implantation period was 57. Post implant patients had 0.47 times the rate of heart failure hospitalizations compared to pre-implant patient (95% CI: 0.31, 0.70; p-value Conclusion In the largest patient population at a VA real-world setting, the implantation of a CardioMEMS results in decreased pulmonary artery pressures and decreased heart failure re-admissions. Further studies are needed to confirm these preliminary findings.

Wingless ligands and beta-catenin expression in the rat endometrium: The role of Wnt3 and Wnt7a/beta-catenin pathway at the embryo-uterine interface.

Wnt/beta-catenin signaling may play an essential role in endometrial decidualization, placentation, and the establishment of pregnancy. We investigate here the possible roles, immunolocalizations, and synthesis of the Wnt3, Wnt7a, and beta-catenin proteins in the rat endometrium during the estrous cycle and early postimplantation period. Wnt3 and Wnt7a had a similar localization and dynamic expression relative to the endometrial stages. Wnt7a immunostaining was not limited only to the luminal epithelial cells, but also to strong stainings in the stromal and endothelial cells. Wnt3, Wnt7a, and beta-catenin were highly synthesized and colocalized at the trophoblast-decidual interface; and were more obvious in the primary decidual zone, the GTCs, and the ectoplacental cone. Beta-catenin was strongly localized at the borders of the mature decidual cells; however, Wnt3 and Wnt7a immunolocalizations were decreased in those cells. As such, the immunolocalization of Wnt3, Wnt7a, and beta-catenin shifted with decidualization and placentation. The expression level of Wnt3, Wnt7a, and beta-catenin messenger RNAs increased in early pregnancy, and especially between Days 8.5 and 9.5. The dramatic changes in the expression of Wnt3, Wnt7a, and beta-catenin observed during the early days of pregnancy and the estrous cycle may indicate their roles in decidualization, stromal cell proliferation, and trophoblast invasion.

Toxicological potential of penconazole on early embryogenesis of white mice Mus musculus in either pre- or post-implantation exposure.

The present investigation was conducted to evaluate the effect of penconazole (PEN) fungicide on early embryogenesis of white mice. In the first experiment, 48 pregnant females were divided into different groups; the first group is control (G1). The second group (G2) was treated daily with PEN (30-, 20-, 10-, 5-mg/kg BW). The third group (G3) was treated with PEN (5-mg/kg BW; day after the other day). The fourth group (G4) was treated with PEN (2.5-mg/kg BW daily) during pre-implantation stage (from the 1st to the 4th day of gestation). The fifth group (G5) was treated with PEN (2.5-mg/kg BW daily) during post-implantation (from the 5th to the 8th day of gestation). The pregnant females were sacrificed at the 14th day of gestation. In the second experiment, 63 pregnant females were classified into control, PEN-treated during pre-implantation period (2.5-mg/kg BW), and PEN-administered during post-implantation period (2.5-mg/kg BW). Each group was sacrificed at stages E6.5, E7.5, E8.5, E9.5, E11.5, E14.5, and E18.5. The high doses of PEN in the first experiment showed failed pregnancy, foetoresorption, and embryo disorganization. High doses of PEN induce alterations in the uterus tissue at the level of histology and immunohistochemistry for the expression of TGFβ2, TNFR2, Caspase 10, and HSP70. The low doses of PEN in the second experiment showed upregulated expression of TGFβ2, TNFR2, Caspase 10, and HSP70 at stages E6.5 and E7.5. In conclusion, PEN was found to alter the suitable uterine environment for proper implantation and development at the levels of histological and immunohistochemical that could create a risk during the full course of embryogenesis.

DNA methylation analysis of multiple imprinted DMRs in Sotos syndrome reveals IGF2-DMR0 as a DNA methylation-dependent, P0 promoter-specific enhancer.

Haploinsufficiency of NSD1, which dimethylates histone H3 lysine 36 (H3K36), causes Sotos syndrome (SoS), an overgrowth syndrome. DNMT3A and DNMT3B recognizes H3K36 trimethylation (H3K36me3) through PWWP domain to exert de novo DNA methyltransferase activity and establish imprinted differentially methylated regions (DMRs). Since decrease of H3K36me3 and genome‐wide DNA hypomethylation in SoS were observed, hypomethylation of imprinted DMRs in SoS was suggested. We explored DNA methylation status of 28 imprinted DMRs in 31 SoS patients with NSD1 defect and found that hypomethylation of IGF2‐DMR0 and IG‐DMR in a substantial proportion of SoS patients. Luciferase assay revealed that IGF2‐DMR0 enhanced transcription from the IGF2 P0 promoter but not the P3 and P4 promoters. Chromatin immunoprecipitation‐quantitative PCR (ChIP‐qPCR) revealed active enhancer histone modifications at IGF2‐DMR0, with high enrichment of H3K4me1 and H3 lysine 27 acetylation (H3K27ac). CRISPR‐Cas9 epigenome editing revealed that specifically induced hypomethylation at IGF2‐DMR0 increased transcription from the P0 promoter but not the P3 and P4 promoters. NSD1 knockdown suggested that NSD1 targeted IGF2‐DMR0; however, IGF2‐DMR0 DNA methylation and IGF2 expression were unaltered. This study could elucidate the function of IGF2‐DMR0 as a DNA methylation dependent, P0 promoter‐specific enhancer. NSD1 may play a role in the establishment or maintenance of IGF2‐DMR0 methylation during the postimplantation period.

Leaf ethanolic extract of Etlingera hemesphaerica Blume alters mercuric chloride teratogenicity during the post-implantation period in Mus musculus.

This study aimed to investigate the effectiveness of leaf ethanolic extract of Etlingera hemisphaerica (LE3H) against the teratogenic effects of mercuric chloride (HgCl2) in mice (Mus musculus). Pregnant M. musculus were divided into four groups, each consisting of 10 dams, and received drink and food ad libitum. The first, second, and third, and fourth (control) groups were administered with LE3H, HgCl2, HgCl2 + LE3H, and double-distilled water alone, respectively. HgCl2 (5mg/kg bw) was administered by injection on gestation day (GD) 9, and LE3H (0.39mg/g bw) was administered by gavage on GD 10. Treated and control animals were killed by cervical dislocation on GD 18, dissected, and the fetuses were collected for evaluation of maternal, embryonic, and fetal toxicity. Eight parameters were measured: (a) embryo resorption or resorbed embryo, (b) dead fetus, (c) living fetus, (d) morphologically normal living fetus, (e) malformed living fetus, (f) number of MLF, (g) length of MNLF, and (h) weight of MNLF. LE3H caused 4 (50.00%), whereas HgCl2 resulted in 7 (87.50%) parameters that were significantly different from those of the control, indicating that the teratogenicity of HgCl2 was significantly higher than that of LE3H. HgCl2 + LE3H showed two effects of LE3H on the teratogenicity of HgCl2: increased 2 (25.00%), and decreased 6 (75.00%). Thus, LE3H decreased the teratogenic effects of HgCl2 in M. musculus.

The STARflo™ glaucoma implant: a single-centre experience at 24months.

To evaluate the 24-month outcomes following STARflo™ implantation in patients with moderate or advanced open-angle glaucoma. We enrolled 32 patients (40 eyes) with high intraocular pressure (IOP) resistant to topical and systemic medical therapy. After baseline assessments, patients were implanted with STARflo™ implants with the goal of IOP reduction and long-term maintenance. Patients were followed for 24months. Complete success of implantation was defined as a restoration of normal IOP without topical glaucoma medications, while qualified success was defined as a restoration of normal IOP with implantation and topical glaucoma medications. STARflo™ did not satisfactorily reduce IOP at 24months. Twenty-eight eyes (70%) had elevated IOP at least once during the 24months post-implantation period. Five eyes (12.5%) developed corneal decompensation. The average IOP 24months after the surgery was 13.42 ± 6.03mmHg and was not significantly different than IOP at 12months (13.2 ± 5.59mmHg). Moreover, 45% of treated eyes needed additional glaucoma procedures after 1year and 2years to achieve these IOPs. STARflo™ did reduce the average number of topical glaucoma medications from 2.7 pre-implantation to 1.17 at 24months postoperatively in the rest 55% of the eyes. The STARflo™ implantation did not meet success criteria and so appears to be an ineffective alternative to filtering surgical procedures for patients with treatment-refractory open-angle glaucoma.

The influence of cochlear implantation on aggression behaviors in children

ObjectiveIt has been suggested that children with higher degree of hearing loss may show more severe externalizing (e.g., aggression) and internalizing (e.g., depression and anxiety) behavioral problems compared to that of their normally hearing peers. The purpose of this study was to investigate the influence of sound amplification through cochlear implants (CIs) on aggressive behaviors in Persian children. MethodsDuring a prospective study design, 72 children (40 girls, 32 boys) who underwent unilateral implantation and 72 (40 girls, 32 boys) age-matched normally hearing children (control group) participated. All CI children had age of <4 years at the time of implantation. The Aggression Scale for Preschoolers (ASFP) was utilized to measure various types of aggression including verbal aggression, physical aggression, relational aggression, and impulsive anger. The speech performance of children was also evaluated using Speech Intelligibility Rating (SIR). The ASFP and SIR measurements were conducted at the pre-implantation phase and at 6, 12, and 18 months post-implantation period. ResultsA repeated measures analysis of variance showed significant reduction in ASFP scores from “pre-operation” to “post-operative” conditions in CI group. However, aggression level in CI recipients were significantly higher than children in control group. In the analyses of ASFP subscales, we found a reduction and a consequent improvement in verbal aggression, physical aggression and impulsive anger. However, no significant difference was found in relational aggression across various time intervals. Furthermore, children who received CI before 2 years old exhibited better aggression level than children who received their devices at later ages. The significantly negative correlation (r = −0.76, p < 0.001) was found between the SIR and ASFP scores, so that lower speech intelligibity ability scores were associated with more aggressive behavior problems. ConclusionThis study suggests that young children with noticeable degree of hearing loss may experience higher level of aggression compared to normally hearing peers. However, when they undergo early implantation and attend at regular post-operative rehabilitation programs, their aggression status will improve seriously.