Plant-based extracts are rich sources of phenolic compounds, which may act as skin antiaging mediators. Herein, Cistus albidus L. (Ca), Cistus ladanifer L. subsp. ladanifer (Cl) and Cistus salviifolius L. (Cs) were selected to test whether their phytochemical profile and bioactive potential align to target human skin aging. Hydroethanolic extracts (HEs) were prepared and characterized using infrared vibrational spectroscopy (FTIR-ATR) and liquid chromatography-mass spectrometry (LC-MS). Non-toxic concentrations were screened, and cytoprotective and antioxidant effects were studied in tert-butyl hydroperoxide-stimulated normal human dermal fibroblasts (NHDFs). Lipopolysaccharide-stimulated RAW 264.7 macrophages were used to assess anti-inflammatory activity, the Organization for Economic Co-operation and Development (OECD) Test Guideline No. 439 was used to assess irritant effects, and the anti-senescence potential was assessed in etoposide-stimulated NHDFs. A series of enzymatic inhibition assays was performed. All extracts comprised ellagic acid derivatives, as well as myricetin and quercetin derivatives in Cs and Ca. The HE of Cs was also markedly composed of ligstroside. At non-toxic concentrations, cytoprotective effects were observed in NHDFs. However, only Cs and Cl exhibited significant antioxidant activity in these cells (p < 0.001 and p < 0.0001, respectively). In addition to that, Cl demonstrated highly significant anti-inflammatory (p < 0.0001) and anti-senescence (p < 0.0001) effects. Cs and Cl showed a remarkable potential to inhibit elastase; in addition, Cs also showed anti-hyaluronidase and anti-tyrosinase activities. Meaningfully, Cs and Cl extracts did not exhibit skin irritant effects. The unveiled potential of Cl in skin aging offset highlights the need to elucidate the detailed mechanisms of action, paving the way for the development of skin anti-aging formulations.

Emerging resistance in malaria vectors and parasites has accelerated the development of novel control strategies such as the Sterile Insect Technique and Wolbachia-based approaches, which require large-scale mosquito rearing. These methods traditionally rely on vertebrate Blood, raising ethical and logistical concerns. This study evaluated the effects of a BLOODless™ on wing size, longevity, and Plasmodium falciparum susceptibility in Anopheles coluzzii and An. gambiae from Burkina Faso, and the Kisumu An. gambiae strain. Blood and Bloodless-reared mosquitoes were compared across seven generations. Wing size was unaffected by diet in the 3rd and 7th generations, though sex impacted size in the 3rd generation only. A diet-species interaction was detected in the 7th generation. Plasmodium falciparum infection rates were similar in the 3rd generation, but by the 6th, Blood-fed mosquitoes showed higher oocyst prevalence, though intensity remained unchanged. Infection outcomes were species-dependent only at this later stage. Longevity, initially comparable across groups, increased significantly in Bloodless-fed mosquitoes by the 7th generation. Species had no effect on survival. A Blood-free diet enables sustained mosquito rearing while preserving traits essential for malaria research, providing an ethical, effective alternative to vertebrate Blood in vector control and experimental programs.

OBJECTIVE:To compare the long-term microtensile bond strength (μTBS) to coronal dentin using pre-endodontic dentin sealing (PEDS) and post-endodontic adhesion (PEA) techniques under various endodontic irrigation protocols. MATERIALS AND METHODS:Ten study groups (n = 10) were established based on the timing of adhesive application (PEDS versus PEA) and irrigation protocol: distilled water (control), 3% sodium hypochlorite (NaOCl), 3% NaOCl followed by 17% ethylenediaminetetraacetic acid (EDTA), 3% NaOCl followed by 17% EDTA and 2% chlorhexidine, and a mixture of 3% NaOCl and 9% etidronic acid (HEDP). Specimens underwent μTBS testing after a six-month microspecimen aging period. Fracture patterns were analyzed, and adhesive interfaces were assessed using scanning electron microscopy (SEM). Statistical analysis employed a mixed linear regression model with a 5% significance level. RESULTS:PEDS consistently preserved high bond strength across all irrigation protocols (57.4-59.5 MPa), while PEA groups treated with endodontic irrigants resulted in significantly lower values (33.3-40.8 MPa; p < 0.001). No significant differences were observed within the PEDS groups (p > 0.05). SEM analysis revealed consistent hybrid layers in PEDS and PEA/Control groups, while PEA groups treated with endodontic irrigation solutions showed significant resin-dentin interface variations and interfacial gaps. CONCLUSIONS:The PEDS technique preserved high and consistent μTBS regardless of the irrigation protocol, whereas endodontically irrigated PEA groups exhibited significantly reduced bond strength. PEDS offers a predictable approach to optimizing adhesive performance in endodontic-restorative treatments. CLINICAL SIGNIFICANCE:Integrating PEDS into routine endodontic-restorative workflow is recommended to enhance long-term bond strength to coronal dentin. The PEDS technique ensures consistent adhesive performance regardless of the endodontic irrigation protocol, enhancing restorative predictability and treatment success while preserving tooth structure.

Human oocytes are highly specialized cells with the capacity to store and regulate mRNAs during oocyte maturation, in preparation for post-fertilization steps. Here we performed single-oocyte transcriptomic analysis of human oocytes in three meitoic maturation stages - Germinal Vesicle (GV; n = 6), Metaphase I (MI; n = 6) and Metaphase II (MII; n = 7). Single-oocyte transcriptomic analysis revealed that the total number of expressed genes progressively decreased from GV to MII stages, with 9660 genes being transcribed in GV, 8734 in MI and 5889 in MII. The same tendency was observed for the number of uniquely expressed genes, with 1328 uniquely expressed genes in GV, 401 in MI and 72 in MII. GO analysis of the uniquely expressed genes showed distinct terms in GV oocytes such as transferase activity, organonitrogen compound metabolic process and ncRNA processing. Analysis of Differentially Expressed Genes (DEGs) between the three maturation stages revealed 1165 DEGs between GV and MII oocytes, with 635 being upregulated and 528 downregulated, 42 DEGs between GV and MI, with 38 being upregulated and 4 downregulated, and no significant changes in gene expression between MI and MII oocytes. Comprehensive analysis of epigenetic regulators showed high expression of several histone-modifying enzymes, namely deacetylases, acetylases, lysine demethylases and methyltransferases, and DNA methylation regulators, namely the maintenance methyltransferase DNMT1 and its co-regulators DPPA3 and UHRF1. Some of these epigenetic regulators were differentially expressed between maturation stages, namely SIRT3, SIRT6, KDM3AP1, KMT2E, DNMT1, DPPA3 and the MEST and RASGRF1 imprinted genes. Our study contributes with important information on the transcriptional landscape of human oocytes in different stages of meiotic maturation, providing important insights into candidate biomarkers of human oocyte quality.

Palladium-based anticancer drugs are promising alternatives to platinum agents, with potential to overcome acquired resistance and reduce side effects. Among them, the dinuclear Pd(II)-spermine complex (Pd2Spm) has shown notable cytostatic activity in vitro. However, its synthesis mechanism and structural properties remain poorly understood, which hinders further development. This study reports an optimised synthetic route for Pd2Spm, revealing pH dependence and temperature-induced isomeric changes. These variations were thoroughly characterised by vibrational spectroscopy, leading to the identification of the pure diastereomer (R,S), the enantiomeric mixture ((R,R) and (S,S)) and the mixture containing these three isomers. Cell viability tests were carried out in the human triple negative breast cancer MDA-MB-231 cell line for the diastereomer and the enantiomers and diastereomer mixture, demonstrating equivalent results for these entities (IC50 equal to 2.7 and 2.6 μM at 72 h, respectively). This is the first report of an optimised synthesis and anticancer in vitro assays for a well-defined Pd2Spm structure. The present findings strengthen the potential of Pd2Spm as a next-generation anticancer metallodrug, paving the way for preclinical trials.

OBJECTIVE:To evaluate the effects of four endodontic irrigation protocols on the chemical composition and ultrastructure of coronal dentin. MATERIALS AND METHODS:Coronal dentin fragments were assigned to five study groups: NaOCl (3% sodium hypochlorite), NaOCl/EDTA (3% NaOCl and 17% ethylenediaminetetraacetic acid), NaOCl/EDTA/CHX (3% NaOCl, 17% EDTA and 2% chlorhexidine), NaOCl/HEDP (mixture of 3% NaOCl and 9% etidronic acid), and control (distilled water). Confocal Raman microscopy was employed to analyze the spatial distribution of organic and inorganic components, while attenuated total reflection Fourier transform infrared spectroscopy and energy-dispersive spectroscopy were used to assess the surface composition of dentin. Ultrastructural evaluation was conducted using scanning electron microscopy (SEM). Statistical analysis was performed using a mixed linear model with a significance level of 0.05. RESULTS:All NaOCl-treated groups showed reduced amide II (p < 0.001), indicating protein degradation. Exclusive NaOCl irrigation yielded the lowest amide II, highest mineral content, and increased phosphate/amide II and carbonate/phosphate ratios (p < 0.05). Chelators reduced mineral content (p < 0.001), with NaOCl/HEDP and NaOCl/EDTA/CHX producing more mineralized surfaces than NaOCl/EDTA and control groups (p < 0.05). A general decrease in organic (C and N) and an increase in inorganic (O, P, and Ca) components occurred across treatments, particularly in NaOCl and NaOCl/HEDP groups. EDTA disturbed the Ca/P equilibrium (p < 0.05). SEM showed a dense smear layer and mostly obliterated tubules in NaOCl and control samples, while chelators reduced the smear layer, partially opened tubules, and caused erosion. CONCLUSIONS:Exclusive use of 3% NaOCl produces hypermineralized, collagen-depleted coronal dentinal surfaces, with a dense smear layer. Sequential irrigation with 17% EDTA induces stronger demineralization compared to a continuous chelation protocol with 9% HEDP. Both EDTA- and HEDP-treated coronal dentin display partially opened dentinal tubules, signs of erosion, and substantial smear layer reduction. CLINICAL SIGNIFICANCE:Clinically recommended endodontic irrigation protocols significantly alter the chemical composition and ultrastructural integrity of coronal dentin, the primary substrate for adhesive restorations. These findings enhance the understanding of post-irrigation coronal dentin conditions and their potential implications on the interaction with adhesive restorative materials.

Recurrent pregnancy loss is a reproductive disorder affecting about 1 to 5 % of pregnant women worldwide that requires our attention, especially considering that about 50 % of cases are idiopathic. The present study is focused on testing a possible association between extreme skewed X-chromosome inactivation patterns and/or shortened telomeres with idiopathic cases since both are considered non-consensual potential causes underlying recurrent pregnancy loss in the scientific community. For this purpose, two groups of women were analyzed and compared: a group of women with idiopathic recurrent pregnancy loss and a second group of age-matched women with proven fertility, and both X-chromosome inactivation patterns and telomere length were measured and compared from maternal DNA extracted from peripheral blood. Our data showed no statistically significant differences between groups, suggesting no association between extreme skewed X-chromosome inactivation or shortened telomeres with recurrent pregnancy losses. Additionally, the effect of maternal age on both X-chromosome inactivation pattern and telomere length was tested, but no significant correlation was observed between advanced maternal age and extreme skewed X-chromosome inactivation or telomere shortening. This study represents one more valid contribution to the investigation of causes underlying recurrent pregnancy loss suggesting that, new variables may be considered since the pattern of X-chromosome inactivation and telomere length do not seem to be related to this reproductive disorder. Briefly, considering its clinical relevance, it is mandatory a continuous effort in the scientific community to cover new potential recurrent pregnancy loss-related causes.

Abstract Background The current rise of new innovative tools for mosquito control, such as the release of transgenic mosquitoes carrying a dominant lethal gene and Wolbachia-based strategies, necessitates a massive production of mosquitoes in the insectary. However, currently laboratory rearing depends on vertebrate blood for egg production and maintenance. This practice raises ethical concerns, incurs logistical and cost limitations, and entails potential risk associated with pathogen transmission and blood storage. Consequently, an artificial blood-free diet emerges as a desirable alternative to address these challenges. This study aims to evaluate the effects of a previously formulated artificial blood-free diet (herein referred to as BLOODless) on Anopheles gambiae (An. gambiae s.s.; IFAKARA) gonotrophic parameters and fitness compared with bovine blood. Methods The study was a laboratory-based comparative evaluation of the fitness, fecundity and fertility of An. gambiae s.s. (IFAKARA) reared on BLOODless versus vertebrate blood from founder generation (F0) to eighth generation (F8). A total of 1000 female mosquitoes were randomly selected from F0, of which 500 mosquitoes were fed with bovine blood (control group) and the other 500 mosquitoes were fed with BLOODless diet (experimental group). The feeding success, number of eggs per female, hatching rate and pupation rate were examined post-feeding. Longevity and wing length were determined as fitness parameters for adult male and female mosquitoes for both populations. Results While blood-fed and BLOODless-fed mosquitoes showed similar feeding success, 92.3% [95% confidence interval (CI) 89.7–94.9] versus 93.6% (95% CI 90.6–96.6), respectively, significant differences emerged in their reproductive parameters. The mean number of eggs laid per female was significantly higher for blood-fed mosquitoes (P < 0.001) whereas BLOODless-fed mosquitoes had significantly lower hatching rates [odds ratio (OR) 0.17, 95% CI 0.14–0.22, P < 0.001]. Wing length and longevity were similar between both groups. Conclusions This study demonstrates the potential of the BLOODless diet as a viable and ethical alternative to vertebrate blood feeding for rearing An. gambiae s.s. This breakthrough paves the way for more efficient and ethical studies aimed at combating malaria and other mosquito-borne diseases. Graphical Abstract

In recent years, there has been increasing awareness of the preservation, protection and sustainable use of natural resources.Water resources, being one of the most important natural resources, face major threats due to contamination by pollutants of various types and origins.Maintaining the quality of water resources requires more robust, reliable and more frequent monitoring than traditional techniques of data collection based on sporadic, discontinuous and manual processes.The management of large geographical areas, the insufficient spatiotemporal discretization of the values of samples collected by traditional processes and the unpredictability of natural phenomena, require a new approach to data collection procedures.This article, which is the result of ongoing research, defines the technical requirements and technologies used in a continuous and regular monitoring of surface water quality in freshwater systems, whose data acquisition system helps to identify the sources of pollution and the contaminants flow along the waterways.The design of a versatile real-time water quality monitoring system, which, due to its environmental constraints should be based on renewable energies and wireless transfer of energy, will contribute to improve the management and effective protection of water resources.

Mosquito-borne diseases kill millions of people each year. Therefore, many innovative research and population control strategies are being implemented but, most of them require large-scale production of mosquitoes. Mosquito rearing depends on fresh blood from human donors, experimentation animals or slaughterhouses, which constitutes a strong drawback since high blood quantities are needed, raising ethical and financial constraints. To eliminate blood dependency and the use of experimentation animals, we previously developed BLOODless, a patented diet that represents an important advance towards sustainable mosquito breeding in captivity. BLOODless diet was used to maintain a colony of Anopheles stephensi for 40 generations. Bloodmeal appetite, fitness, Plasmodium berghei infectivity, whole genome sequencing and microbiota were evaluated over time. Here we show that BLOODless can be implemented in Anopheles insectaries since it allows long-term rearing of mosquitoes in captivity, without a detectable effect on their fitness, infectivity, nor on their midgut and salivary microbiota composition.

Brazil is the largest producer in the world of the species Agave sisalana, sisal. The residue of the sisal, which is the result of the extraction of fibers from its leaves, represents 95% of its weight. Considering that sisal leaves have high concentrations of sapogenins and aiming at a future phytotherapeutic, in this study, the alcoholic fraction of sisal, AFS, was developed, and the sapogenins were characterized. In vitro, the cytotoxicity (MTT) and the anti-inflammatory effect of AFS (phagocytosis and hemolysis inhibition) were evaluated. In vivo, the analgesic (formalin test—FT) and anti-inflammatory (paw edema test—PET) activities of AFS, orally, and the cream containing AFS, topical, were analyzed. The results demonstrated that AFS contains hecogenin and tigogenin and is not cytotoxic. In vitro, 0.5, 1, and 2 mg/mL of AFS showed anti-inflammatory activity similar to the positive control (PC). In the FT, the dose of 25 mg/kg did not differ from the PC in the neurogenic phase (p > 0.05). In the PET, 25 and 50 mg/kg of AFS differed from the negative control (NC) (p < 0.05), and the cream with AFS (5 mg/g) showed activity similar to the PC. The therapeutic activities of AFS probably result from sapogenins. In the future, we expect to develop an anti-inflammatory from the thousands of tons of sisal waste discarded in Brazil.

Numerous therapeutic and diagnostic approaches used within a clinical setting depend on the administration of compounds via systemic delivery. Biomaterials at the nanometer scale, as dendrimers, act as delivery systems by improving cargo bioavailability, circulation time, and the targeting of specific tissues. Although evaluating the efficacy of pharmacological agents based on nanobiomaterials is crucial, conducting toxicological assessments of biomaterials is essential for advancing clinical translation. Here, a zebrafish larvae model was explored to assess the biocompatibility of poly(amido amine) (PAMAM), one of the most exploited dendrimers for drug delivery. We report the impact of a systemic injection of polyethylene glycol (PEG)-modified G4 PAMAM conjugated with rhodamine (Rho) as a mimetic drug (PEG-PAMAM-Rho) on survival, animal development, inflammation, and neurotoxicity. A concentration- and time-dependent effect was observed on mortality, developmental morphology, and innate immune system activation (macrophages). Significant effects in toxicological indicators were reported in the highest tested concentration (50 mg/mL PEG-PAMAM-Rho) as early as 48 h post-injection. Additionally, a lower concentration of PEG-PAMAM-Rho (5 mg/mL) was found to be safe and subsequently tested for neurotoxicity through behavioral assays. In accordance, no significative signs of toxicity were detected. In conclusion, the dose response of the animal was assessed, and the safe dosage for future use in theragnostics was defined. Additionally, new methodologies were established that can be adapted to further studies in toxicology using other nanosystems for systemic delivery.

The trophoblast cells are responsible for the transfer of nutrients between the mother and the foetus and play a major role in placental endocrine function by producing and releasing large amounts of hormones and growth factors. Syncytiotrophoblast cells (STB), formed by the fusion of mononuclear cytotrophoblasts (CTB), constitute the interface between the foetus and the mother and are essential for all of these functions. We performed transcriptome analysis of human placental samples from two control groups—live births (LB), and stillbirths (SB) with a clinically recognised cause—and from our study group, idiopathic stillbirths (iSB). We identified 1172 DEGs in iSB, when comparing with the LB group; however, when we compared iSB with the SB group, only 15 and 12 genes were down- and upregulated in iSB, respectively. An assessment of these DEGs identified 15 commonly downregulated genes in iSB. Among these, several syncytiotrophoblast markers, like genes from the PSG and CSH families, as well as ALPP, KISS1, and CRH, were significantly downregulated in placental samples from iSB. The transcriptome analysis revealed underlying differences at a molecular level involving the syncytiotrophoblast. This suggests that defects in the syncytial layer may underlie unexplained stillbirths, therefore offering insights to improve clinical obstetrics practice.