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This study aimed to investigate the biological response induced by hydroxyapatite (HAp) and zinc-doped HAp (ZnHAp) in human gingival fibroblasts and to explore their antimicrobial activity. The ZnHAp (with xZn = 0.00 and 0.07) powders, synthesized by the sol-gel method, retained the crystallographic structure of pure HA without any modification. Elemental mapping confirmed the uniform dispersion of zinc ions in the HAp lattice. The size of crystallites was 18.67 ± 2 nm for ZnHAp and 21.54 ± 1 nm for HAp. The average particle size was 19.38 ± 1 nm for ZnHAp and 22.47 ± 1 nm for HAp. Antimicrobial studies indicated an inhibition of bacterial adherence to the inert substrate. In vitro biocompatibility was tested on various doses of HAp and ZnHAp after 24 and 72 h of exposure and revealed that cell viability decreased after 72 h starting with a dose of 31.25 µg/mL. However, cells retained membrane integrity and no inflammatory response was induced. High doses (such as 125 µg/mL) affected cell adhesion and the architecture of F-actin filaments, while in the presence of lower doses (such as 15.625 µg/mL), no modifications were observed. Cell proliferation was inhibited after treatment with HAp and ZnHAp, except the dose of 15.625 µg/mL ZnHAp at 72 h of exposure, when a slight increase was observed, proving an improvement in ZnHAp activity due to Zn doping.

PURPOSE: In vitro maturation (IVM) is a technology that generates mature oocytes following culture of immature cumulus-oocyte complexes (COC) in vitro. IVM is characterized by minimal patient stimulation, making it attractive for certain patient groups. Recently, a biphasic IVM system, capacitation (CAPA)-IVM, has shown improved clinical outcomes relative to standard IVM; however, it remains less efficient than IVF. This study assessed whether supplementation of CAPA-IVM culture media with the novel TGFβ superfamily proteins cumulin and super-GDF9 improves subsequent mouse embryo development. METHODS: Immature mouse COCs were cultured by standard IVM or biphasic IVM ± cumulin or super-GDF9. RESULTS: Both cumulin and super-GDF9 in standard IVM significantly improved day-6 blastocyst rate (53.9% control, 73.6% cumulin, 70.4% super-GDF9; p = 0.006; n = 382–406 oocytes). Cumulin or super-GDF9 in CAPA-IVM did not alter embryo yield or blastocyst cell allocation in an unstimulated model. Moreover, cumulin did not alter these outcomes in a mild PMSG stimulation model. Cumulin in CAPA-IVM significantly increased cumulus cell expression of cumulus expansion genes (Ptgs2, Ptx3, Adamts1, Gfat2) and decreased Lhr expression relative to control. However, cumulin-induced mRNA expression of cumulus cell (Ptgs2, Ptx3) and oocyte genes (Gdf9, Bmp15, Oct4, Stella) in CAPA-IVM remained significantly lower than that of in vivo matured cells. CONCLUSION: Cumulin did not provide an additional beneficial effect in biphasic IVM in terms of blastocyst yield and cell allocation; however in standard IVM, cumulin and super-GDF9 significantly improve oocyte developmental competence.

In vitro maturation (IVM) is an alternative assisted reproductive technology (ART) with reduced hormone related side-effects and treatment burden compared to conventional IVF. Capacitation (CAPA)-IVM is a biphasic IVM system with improved clinical outcomes compared to standard monophasic IVM. Yet, CAPA-IVM efficiency compared to conventional IVF is still suboptimal in terms of producing utilizable blastocysts. Previously we have shown that CAPA-IVM leads to a precocious increase in cumulus cell (CC) glycolytic activity during cytoplasmic maturation. In the current study, considering the fundamental importance of CCs for oocyte maturation and cumulus-oocyte complex (COC) microenvironment, we further analyzed the bioenergetic profiles of maturing CAPA-IVM COCs. Through a multi-step approach, we (i) explored mitochondrial function of the in vivo and CAPA-IVM matured COCs through real-time metabolic analysis with Seahorse analyzer; and to improve COC metabolism (ii) supplemented the culture media with lactate and/or super-GDF9 (an engineered form of growth differentiation factor 9) and (iii) reduced culture oxygen tension. Our results indicated that the pre-IVM step is delicate and prone to culture related disruptions. Lactate and/or super-GDF9 supplementations failed to eliminate pre-IVM induced stress on COC glucose metabolism and mitochondrial respiration. However, when performing pre-IVM culture under 5% oxygen tension, CAPA-IVM COCs showed a similar bioenergetic profiles compared to in vivo matured counterparts. This is the first study providing real-time metabolic analysis of the COCs from a biphasic IVM system. The currently used analytical approach provides the quantitative measures and the rational basis to further improve IVM culture requirements.

Establishing an ideal human follicle culture system for oncofertility patients relies mainly on animal models since donor tissue is scarce and often of suboptimal quality. The in vitro system developed in our laboratory supports the growth of prepubertal mouse secondary follicles up to mature oocytes. Given the importance of glucose in preparing the oocyte for proper maturation, a baseline characterization of follicle metabolism both in the culture system and in vivo was carried out. Markers of glucose-related pathways (glycolysis, tricarboxylic acid [TCA] cycle, pentose phosphate pathway [PPP], polyol pathway, and hexosamine biosynthetic pathway), as well as the antioxidant capacity, were measured in the different follicle cell types by both enzymatic activities (spectrophotometric detection) and gene expression (qPCR). This study confirmed that in vivo the somatic cells, mainly granulosa, exhibit intense glycolytic activity, while oocytes perform PPP. Throughout the final maturation step, oocytes in vivo and in vitro showed steady levels for all the key enzymes and metabolites. On the other hand, ovulation triggers a boost of pyruvate and lactate uptake in cumulus cells in vivo, consumes reduced nicotinamide adenine dinucleotide phosphate, and increases TCA cycle and small molecules antioxidant capacity activities, while in vitro, the metabolic upregulation in all the studied pathways is limited. This altered metabolic pattern might be a consequence of cell exhaustion because of culture conditions, impeding cumulus cells to fulfill their role in providing proper support for acquiring oocyte competence.

In vitro maturation (IVM) is an assisted reproduction technique with reduced hormone-related side-effects. Several attempts to implement IVM in routine practice have failed, primarily due to its relatively low efficiency compared with conventional in vitro fertilization (IVF). Recently, capacitation (CAPA)-IVM—a novel two-step IVM method—has improved the embryology outcomes through synchronizing the oocyte nuclear and cytoplasmic maturation. However, the efficiency gap between CAPA-IVM and conventional IVF is still noticeable especially in the numerical production of good quality embryos. Considering the importance of glucose for oocyte competence, its metabolization is studied within both in vivo and CAPA-IVM matured mouse cumulus-oocyte-complexes (COCs) through direct measurements in both cellular compartments, from transcriptional and translational perspectives, to reveal metabolic shortcomings within the CAPA-IVM COCs. These results confirmed that within in vivo COC, cumulus cells (CCs) are highly glycolytic, whereas oocytes, with low glycolytic activity, are deviating their glucose towards pentose phosphate pathway. No significant differences were observed in the CAPA-IVM oocytes compared with their in vivo counterparts. However, their CCs exhibited a precocious increase of glycolytic activity during the pre-maturation culture step and activity was decreased during the IVM step. Here, specific alterations in mouse COC glucose metabolism due to CAPA-IVM culture were characterized using direct measurements for the first time. Present data show that, while CAPA-IVM CCs are able to utilize glucose, their ability to support oocytes during final maturation is impaired. Future CAPA-IVM optimization strategies could focus on adjusting culture media energy substrate concentrations and/or implementing co-culture strategies.Summary sentence Pre-maturation culture conditions for immature oocytes from small antral follicles affect glycolytic activity and antioxidant defense capacity in mouse CCs during IVM.

Study question Are there significant differences in carbohydrate metabolism trends between in vivo and in vitro grown mouse antral follicles during oocyte final maturation? Summary answer Glucose metabolism characterization during GV to MII transition revealed altered metabolic patterns mainly in cumulus cells of in vitro grown and matured mouse antral follicles. What is known already For some cancer patients fertility restoration is dependent on using efficient in vitro follicle culture systems. As human donor ovarian tissue available for research is limited, establishing such culture systems relies on data generated from animal models. The culture system previously developed in our laboratory supports in vitro growth of mouse preantral follicles with good oocyte maturation rates but lower developmental competence compared to in vivo grown oocytes. Tracking and comparing the metabolic changes after meiotic maturation in in vitro and in vivo follicles could serve as a screening tool for improving culture conditions and identifying metabolic quality markers. Study design, size, duration Mouse secondary follicle culture was performed. In vitro grown oocytes, their corresponding cumulus (CC) and granulosa cells (GC) were collected from antral follicles, at germinal vesicle stage (GV) on day 9, and at metaphase 2 (MII) on day 10, after hCG/EGF stimulation. In vivo age-matched controls were obtained after intraperitoneal injections with eCG for GV, or with eCG and hCG for MII. In vivo GC after ovulation were not included. Participants/materials, setting, methods Glucose metabolism trends were compared during final maturation between in vitro grown antral follicles and their in vivo controls. Follicles that failed to resume meiosis in vitro were also included. Enzymatic spectrophotometric assays were used to measure glycolysis, pentose phosphate pathway (PPP), tricarboxylic acid (TCA) cycle, and the antioxidant capacity in individual cell types. Pools of 5 oocytes and corresponding somatic cells were collected, from 3 independent experiments. Unpaired t-test was performed with significance when p Main results and the role of chance Important differences were detected between in vivo and in vitro conditions. GV to MII transition in in vivo follicles leads to a metabolic boost in CC as indicated by: i. significant increase in glycolysis, PPP and TCA cycle activity; ii. higher total antioxidant capacity (TAC) (p Meiotic maturation triggered no significant differences in any of the metabolic pathways for in vitro grown oocytes. Contrary to their in vivo controls, in vitro CC showed significant upregulations limited to aconitase, lactate dehydrogenase (LDH) and glutathione-s-transferase (GST) activity (p Significant differences were detected between in vivo GV follicles and the in vitro failed-to-mature ones. Oocytes from impaired follicles have higher NADP+ levels (p Limitations, reasons for caution The altered metabolic patterns described here in in vitro follicles during oocyte GV to MII transition are probably the cumulative effects of both growth and maturation in vitro. Wider implications of the findings: We explored extensively and directly, for the first time, several enzymes and metabolites involved in follicle glucose and redox metabolism in different cell types separately. Understanding of the follicle metabolic requirements is essential for the optimization of follicle culture systems and could lead to development of oocyte quality markers. Trial registration number Not applicable

In vitro oocyte growth is widely studied as an alternative fertility preservation approach. Several animal models are used to generate extensive information on this complex process regulated by the constant and dynamic interaction between the oocyte and its somatic compartment throughout follicle growth and maturation. A two-dimensional attachment mouse secondary follicle culture system was used to assess the oocyte’s capacity to overcome disconnection from its somatic companions at different developmental stages for final competence acquisition. To test this, complete mechanical denudation of oocytes from preantral (PA) and early antral (EA) follicles was performed. Established endpoints were the oocyte’s potential to reconnect with somatic cells and the impact of connectivity disruption on mature oocyte quality. This study proves that oocytes from PA and EA cultured mouse follicles can overcome complete denudation, restoring likely functional transzonal projections with no significant differences in meiotic and developmental competence compared with those from intact cultured follicles. These novel findings constitute good premises for developing successful strategies to rescue human oocyte competence in the context of in vitro culture approaches such as nonhuman chorionic gonadotropin triggered in vitro maturation.

Background and Aims: Due to the increasing number of hepatitis C virus (HCV)-infected patients being treated with direct antiviral agents (DAAs) in Romania, we aimed to conduct a pharmacovigilance study in order to comprehensively evaluate the safety profile for the ombitasvir/paritaprevir/ritonavir and dasabuvir (Om/Pa/Ri+Da) regimen. Methods: A retrospective analysis was conducted on the individual case safety reports (ICSRs), extracted from VigiBase® on 1 st February 2018, which included the Om/Pa/Ri+Da regimen as suspected for causing adverse drug reactions (ADRs). Potential drug-drug interactions (DDIs) were checked for all concomitant medication using the IBM Micromedex® tool. Results: Among the 1,102 ICSRs retrieved, 260 were serious (23.5%). Ribavirin was significantly associated with more ADRs reported per case (on average 5.1 vs. 3.2 ADRs/case, p

According to the EU Energy Poverty Observatory, at present, more than 50 million households in the European Union are experiencing energy poverty as a distinct form of poverty. Energy poverty stems from people’s poor access to energy resources and modern means of ensuring comfort in dwellings. In modern times, a decent standard of living and citizens' health have become criteria measuring the level of energy poverty in a society. The level of essential services such as adequate warmth, cooling, lighting and the energy to power appliances could provide a complete and comprehensive picture of energy poverty, thus having an indirect effect on many policy areas such as health, environment and productivity.

The specialized legislation on the regulation of public services reflected the specifics of each historical period. Public service is a complex notion, subject to evolution that has given rise over time to many contradictory discussions, being at the same time an essential notion for administrative law.Presenting the foundations of democracy, Aristotle outlined suma divisio between civil society, which relates to private interests, and the political society that defends "general utility" and "common good”.

We are talking today about innovation, technology, science and more recently we are talking about ,,smart cities”. The "smart city" phenomenon has developed a lot over the last 20 years. We are talking about intelligent cities along and across the globe, because it is a current subject and a need. The objective of this article is to analyze and define the term "smart city", focusing on definitions, features, global trends, and smart administration in these future cities around the globe.

Background In vitro follicle culture (IFC), as applied in the mouse system, allows the growth and maturation of a large number of immature preantral follicles to become mature and competent oocytes. In the human oncofertility clinic, there is increasing interest in developing this technique as an alternative to ovarian cortical tissue transplantation and to preserve the fertility of prepubertal cancer patients. However, the effect of IFC and hormonal stimulation on DNA methylation in the oocyte is not fully known, and there is legitimate concern over epigenetic abnormalities that could be induced by procedures applied during assisted reproductive technology (ART). Results In this study, we present the first genome-wide analysis of DNA methylation in MII oocytes obtained after natural ovulation, after IFC and after superovulation. We also performed a comparison between prepubertal and adult hormonally stimulated oocytes. Globally, the distinctive methylation landscape of oocytes, comprising alternating hyper- and hypomethylated domains, is preserved irrespective of the procedure. The conservation of methylation extends to the germline differential methylated regions (DMRs) of imprinted genes, necessary for their monoallelic expression in the embryo. However, we do detect specific, consistent, and coherent differences in DNA methylation in IFC oocytes, and between oocytes obtained after superovulation from prepubertal compared with sexually mature females. Several methylation differences span entire transcription units. Among these, we found alterations in Tcf4, Sox5, Zfp521, and other genes related to nervous system development. Conclusions Our observations show that IFC is associated with altered methylation at specific set of loci. DNA methylation of superovulated prepubertal oocytes differs from that of superovulated adult oocytes, whereas oocytes from superovulated adult females differ very little from naturally ovulated oocytes. Importantly, we show that regions other than imprinted gDMRs are susceptible to methylation changes associated with superovulation, IFC, and/or sexual immaturity in mouse oocytes. Our results provide an important reference for the use of in vitro growth and maturation of oocytes, particularly from prepubertal females, in assisted reproductive treatments or fertility preservation.

Additional file 8: Table S1. Summary of all PBAT libraries generated for this study, including the following information: the approximate number oocytes used for each library, number of uniquely mappable reads, mapping efficiency, duplication rate and methylation percentage at CpG, CHG and CHH regions.

Non-steroidal anti-inflammatory drugs (NSAIDs) are an important pharmacological class of drugs used for the treatment of inflammatory diseases. They are also characterized by severe side effects, such as gastrointestinal damage, increased cardiovascular risk and renal function abnormalities. In order to synthesize new anti-inflammatory and analgesic compounds with a safer profile of side effects, a series of 2,6-diaryl-imidazo[2,1-b][1,3,4]thiadiazole derivatives 5a&ndash, l were synthesized and evaluated in vivo for their anti-inflammatory and analgesic activities in carrageenan-induced rat paw edema. Among all compounds, 5c showed better anti-inflammatory activity compared to diclofenac, the standard drug, and compounds 5g, 5i, 5j presented a comparable antinociceptive activity to diclofenac. None of the compounds showed ulcerogenic activity. Molecular docking studies were carried out to investigate the theoretical bond interactions between the compounds and target, the cyclooxygenases (COX-1/COX-2). The compound 5c exhibited a higher inhibition of COX-2 compared to diclofenac.

The currently available assisted reproduction techniques for fertility preservation (i.e.in vitromaturation (IVM) andin vitrofertilization) are insufficient as stand-alone procedures as only few reproductive cells can be conserved with these techniques. Oocytes in primordial follicles are well suited to survive the cryopreservation procedure and of use as valuable starting material for fertilization, on the condition that these could be grown up to fully matured oocytes. Our understanding of the biological mechanisms directing primordial follicle activation has increased over the last years and this knowledge has paved the way toward clinical applications. New multistepin vitrosystems are making use of purified precursor cells and extracellular matrix components and by applying bio-printing technologies, an adequate follicular niche can be built. IVM of human oocytes is clinically applied in patients with polycystic ovary/polycystic ovary syndrome; related knowhow could become useful for fertility preservation and for patients with maturation failure and follicle-stimulating hormone resistance. The expectations from the research on human ovarian tissue and immature oocytes cultures, in combination with the improved vitrification methods, are high as these technologies can offer realistic potential for fertility preservation.

Under the new European regulations, it becomes a necessity and a priority for Romania to take adequate political, legislative and administrative measures to stop the decline of the centralized heat supply systems. The energy efficiency is the most cost-effective and fastest growing method to improve security of energy supply, reduce energy loss and costs, and greenhouse gas emissions which are responsible for global warming. Cogeneration, notably high efficiency cogeneration and the improved performance of the centralized heat supply systems have a key contribution to the improvement of energy efficiency. In this regard, at the Municipality of Bucharest, steps have been taken to achieve the Heat Supply Public Service in Bucharest - production, transport, distribution and supply through an integrated centralized heating supply system (S.A.C.E.T.) (S.A.C.E.T. - C.T.E.S.S. - Centralised Thermal Energy Supply System), aiming to improve the performance of the Public Service in terms of energy and respectively economic efficiency, at the same time creating the basis for the settlement of debts accrued by the Public Service Operator - Bucharest District Heating Company (RADET Bucuresti S.A.), to its heat supplier - S.C. Electrocentrale Bucuresti S.A. (ELCEN) through its subsidiary Electrocentrale Bucuresti.

A solution of (55)Fe was measured by using liquid scintillation counting (LSC)-TDCR method. The measurement was performed within the frame of the CIPM CCRI(II)-K2.Fe-55 comparison during the year 2006. Two computation codes were used for the calculation of detection efficiency, the difference between the results being analyzed. The photomultipliers asymmetry and its influence on the results, as well as the long-term stability are also analyzed. The influence of the variation of kB value over the activity value is compared with the case of (3)H standardization.

A new coincidence system, based on two thin NaI(Tl) crystals, 50×2 mm 2 , with beryllium windows, 1.5 mg cm −2 , positioned approximately symmetrically from the source holder (at various distances) was constructed. The electronic chains of a classical 4 π PC– γ system were utilized. A multichannel analyzer was employed for energy calibration, and an oscilloscope for visualization of the pulses. After calibration, the system was used for the standardization of an 125 I solution. An energy window of 12–95 keV containing the entire spectrum (single and sum peak) was employed, and a simple demonstration of Taylor's equation [1967. X-ray–X-ray coincidence counting methods for the standardization of 125 I and 197 Hg. In: Standardization of Radionuclides: Proceedings of a International Atomic Energy Agency Symposium, Vienna, 1966, pp. 341–353, SM-79/65] for whole spectrum counting, was developed. The method and system were tested during the calibration of a LKB 2104 multi-gamma counter, by comparing the detection efficiency values obtained for this prepared standard solution and that utilized in the CIPM-CCRI(II) key comparison. The efficiency of the equipment was calculated as the mean of the two individual values. The result validates the method and demonstrates the accomplishment of the national traceability assurance in nuclear medicine for 125 I.

The paper presents some recent results of the Radionuclide Metrology Laboratory (RML) from IFIN-HH, concerning the assurance of the international traceability for the measurement of (131)I solution aimed to continue the national chain for nuclear medicine applications in Romania. The following sequences of the traceability are presented: the absolute standardization of solution by the 4pibeta-gamma coincidence, efficiency extrapolation method and participation in the BIPM.RI(II)-K1.I-131 key comparison; and recalibration of the CENTRONIC IG12/20A ionization chamber secondary standard. The participation in the IAEA/SSDL (SSRL) (131)I comparison is also presented.