Search Results (270)

Dendrons, also named dendritic wedges, are a kind of molecular tree, having a branched structure linked to a functional core. The functional core can be used in particular for the functionalization of materials. Different types of dendrons are known, synthesized either by a convergent process, from the external part to the core, or by a divergent process from the core to the external part. Polyphosphorhydrazone (PPH) dendrons are always synthesized by a divergent process, which enables a fine-tuning of both the core function and the external functions. They have been used for the functionalization of diverse materials such as silica, titanium dioxide, gold, graphene oxide, or different types of nanoparticles. Nanocomposites based on materials functionalized with PPH dendrons have been used in diverse fields such as catalysts, chemical sensors, for trapping pollutants, to support cell cultures, and against cancers, as will be emphasized in this review. Full article

Background/Objectives: Tuberculous meningitis is the most severe form of tuberculosis in children, with a high mortality and morbidity rate if it is not diagnosed and treated in a timely manner. The aim of this study is to highlight the challenges associated with establishing a diagnosis of tuberculous meningitis in a child with immunosuppression, given the presence of nonspecific clinical manifestations. Methods: We present the case of a 15-year-old adolescent with systemic lupus erythematosus, on immunosuppressive therapy, who is diagnosed with tuberculous meningoencephalitis presenting the clinical, diagnostic and imaging characteristics, as well as the diagnostic traps and limitations associated with this condition. Antituberculosis therapy was started empirically, because there was no improvement in the clinical status with conventional antibiotic therapy; the diagnosis was established 7 days after the start of the antituberculosis treatment, with the help of an acid-fast bacilli culture from the cerebrospinal fluid. Results: The course of the tuberculous meningoencephalitis was slowly favorable, despite the superimposed COVID-19 infection. Delay in administering immunosuppressive therapy led to the onset of renal and joint manifestations. Conclusions: Tuberculous meningitis is a highly lethal, often underdiagnosed disease with nonspecific clinical and imaging manifestations, which can have a favorable outcome if the diagnosis is established early on and treatment is started promptly. Full article

The domestic cat is the primary reservoir host of three flea-borne Bartonella species, one of which (Bartonella henselae) causes reduced fertility and reproductive failure in experimentally infected cats. Vertical transmission of Bartonella has been documented only in B-cell deficient mice, but not immunocompetent animals. As many free-roaming cats are chronically infected with Bartonella and may be immunocompromised by environmental stress or coinfection, we attempted to isolate Bartonella from the fetal and placental tissues of pregnant queens spayed during trap–neuter–release. Four samples from each tissue (ovary, uterus, fetus, and placenta) were split for direct DNA extraction, liquid culture, and culture on a blood agar plate. Samples from infected queens were inoculated into liquid media and sampled weekly for three weeks for DNA extraction and plating. Bartonella DNA was sequenced directly from 28% (5/18) of the free-roaming queens. For these five queens, liquid enrichment culture was attempted in duplicate for fetal and placental samples. Bartonella clarridgeiae DNA was amplified using qPCR liquid enrichment cultures from the placentas of two cats. These findings suggest that viable Bartonella organisms are present in feline reproductive tissue. Additional studies are needed to assess the transplacental transmission of Bartonella spp. and Bartonella’s influence on fetal development. Full article

Over the last 20 years, poverty has undergone profound changes, even affecting people who were once safe from the processes of impoverishment. Hidden under the guise of normal difficulties considered part of the natural course of life, it has lost its “occasional” connotation and has become “established” in people’s lives, causing new and dangerous trends. The article offers some reflections on how these dynamics have become particularly widespread in Italy, resulting in two types of poverty: cultural poverty, linked to the phenomenon of young NEETs (Not in Employment, Education, or Training), and working poverty. These types of poverty are linked and risk feeding off each other. On the one hand, poor cultural tools force young people to settle for low-paid jobs. On the other hand, poor-quality work, due to poor training, discourages people from pursuing education and training and traps poor workers in a situation of social stagnation. A possible tool to tackle these issues could be Responsible Welfare, which focuses on the person as a unique entity, implementing the resilience of individuals to promote self-esteem while also enhancing relational, social, and community resources. Full article

The possibility of tightly controlling the cellular microenvironment within microfluidic devices represents an important step toward precision analysis of cellular phenotypes in vitro. Microfluidic platforms that allow both long-term mammalian cell culture and dynamic modulation of the culture environment can support quantitative studies of cells’ responses to drugs. Here, we report the design and testing of a novel microfluidic device of simple production (single Polydimethylsiloxane layer), which integrates a micromixer with vacuum-assisted cell loading for long-term mammalian cell culture and dynamic mixing of four different culture media. Finite element modeling was used to predict flow rates and device dimensions to achieve diffusion-based fluid mixing. The device showed efficient mixing and dynamic exchange of media in the cell-trapping chambers, and viability of mammalian cells cultured for long-term in the device. This work represents the first attempt to integrate single-layer microfluidic mixing devices with vacuum-assisted cell-loading systems for mammalian cell culture and dynamic stimulation. Full article

In swine reproduction, immune-mediated mechanisms such as neutrophil extracellular trap (NET) formation can affect sperm function and reduce fertility outcomes. This study evaluated the capacity of antioxidant and reproductive compounds—butylated hydroxytoluene (BHT), prostaglandin E₂ (PGE₂), bovine serum albumin (BSA), and seminal plasma (SP)—to modulate NETosis in co-cultures of swine neutrophils and cryopreserved spermatozoa. NET formation was quantified by nuclear area expansion and validated by digital cytometry and immunofluorescence. BHT (0.5 mM) and PGE₂ (10 µM) produced the most significant inhibitory effects, reducing NETotic cell percentages from 34.5 ± 2.7% (sperm-exposed controls) to 12.2 ± 1.3% and 14.5 ± 2.1%, respectively (p < 0.01). SP at 20% decreased NETosis to 16.8 ± 1.8%, while BSA (0.5%) achieved a moderate reduction to 21.3 ± 2.5%. Flow cytometry revealed reduced peroxynitrite levels in sperm treated with SP and BSA. Two NET phenotypes (aggNETs and sprNETs) were identified. BTS medium enhanced NET formation, whereas DNase I degraded NETs effectively. These findings identify porcine NETosis as a redox-sensitive pathway modulated in vitro, suggesting an immunological role in enhancing sperm preservation for swine artificial insemination. Full article

Free-ranging dogs (Canis lupus familiaris) pose a significant but often overlooked threat to wildlife populations and global conservation efforts while also having the potential to contribute positively to conservation initiatives. As generalist predators and scavengers, these adaptable animals can lead to biodiversity loss through predation, disease transmission, competition, and behavioral disruption of native species. This review synthesizes global studies on their ecological impact, highlighting notable cases of predation on endangered species, such as the markhor (Capra falconeri cashmiriensis) in Pakistan and elephant seals (Mirounga angustirostris) in Mexico, as well as the spread of zoonotic diseases like Echinococcus spp. and canine distemper. A growing concern is hybridization between free-ranging dogs and wild canids. Such genetic mixing can erode local adaptations, reduce genetic purity, and undermine conservation efforts for wild canid populations. Current management strategies—including lethal control, trap–neuter–release, and vaccination—have produced mixed results and face challenges related to data limitations, regional variability, and cultural barriers. This review advocates for integrated, context-specific management approaches that consider ecological, social, and economic dimensions. Future research should prioritize standardized definitions and data collection, long-term evaluation of intervention effectiveness, and the socio-economic drivers of dog–wildlife interactions to develop sustainable solutions for mitigating the multifaceted threats imposed by free-ranging dogs to global diversity. Full article

There is a growing interest in using agri-food by-products and a demand for natural substances that might help maintain healthy honey bee colonies. We investigated a by-product of the wine industry, a grape pomace (GP) of the autochthonous Prokupac grape cultivar from Serbia. A hydroethanolic extract (50% (w/v) ethanol) of GP (Prokupac GP extract) obtained by the pressurized liquid extraction (PLE) method was subjected to qualitative profiling of phenolic composition by liquid chromatography with OrbiTrap Exploris 120 mass spectrometer. Then, the extracts’ antioxidant and redox-modulatory activities were evaluated through Electron Paramagnetic Resonance (EPR) spectroscopy. Finally, the extract’s potential to modulate cellular redox status was evaluated using cultured AmE-711 honey bee cells. The results show that the Prokupac GP extract contains a wide array of flavonoids, anthocyanins, stilbenes, and their various conjugated derivatives and that anthocyanins, particularly malvidin-based compounds, dominate. EPR measurements showed strong scavenging activity against superoxide anion (O₂^•−) and hydroxyl radicals (^•OH), with inhibition efficiencies of 84.37% and 81.81%, respectively, while activity against the DPPH radical was lower (17.75%). In the cell-based assay, the Prokupac GP extract consistently provided strong antioxidant protection and modulated the cellular response to oxidative stress by over 14%. In conclusion, while the Prokupac GP extract demonstrated antioxidant properties and the ability to modulate cellular responses to oxidative stress, in vivo studies on honey bees are required to confirm its efficacy and safety for potential use in beekeeping practice. Full article

Therapeutic gene editing strategies utilize endogenous DNA repair pathways—nonhomologous end joining (NHEJ) or homology-directed repair (HDR)—to introduce targeted genomic modifications. Because HDR is restricted to dividing cells, whereas NHEJ functions in both dividing and non-dividing cells, NHEJ-based approaches are better suited for in vivo gene editing in the largely post-mitotic airway epithelium. Homology-independent targeted insertion (HITI), an NHEJ-based method, offers a promising strategy for cystic fibrosis (CF) gene therapy. Here, we applied HITI to drive the expression of a promoterless reporter through an exon trap strategy in both proliferating airway basal cells and well-differentiated primary airway epithelial cultures derived from transgenic ROSA^mTmG ferrets. We also established a versatile human gene editing reporter (GER) airway basal cell line capable of multipotent differentiation, enabling real-time visualization of editing outcomes and the quantitative assessment of HDR- and NHEJ-based editing efficiencies. Together, these platforms provide easily accessible tools for optimizing genome editing strategies in the respiratory epithelium and advancing clinically relevant delivery strategies for CF gene therapy. Full article

This study investigates the impact of hydroxyapatite (HA) nanoparticles (NPs) on the cellular responses of poly(L-lactide-co-ε-caprolactone) (PLCL) scaffolds in bone tissue engineering applications. Three types of PLCL scaffolds were fabricated, varying in HANPs content. Saos-2 osteoblast-like cells (OBs) and THP-1-derived osteoclast-like cells (OCs) were co-cultured on the scaffolds, and cell proliferation was assessed using the MTS assay. The amount of double-stranded DNA (dsDNA) was quantified to evaluate cell proliferation. Expression levels of OBs and OCs markers were analyzed via quantitative polymerase chain reaction (qPCR) and the production of Collagen type I was visualized using confocal microscopy. Additionally, enzymatic activity of alkaline phosphatase (ALP) and tartrate-resistant acid phosphatase (TRAP or ACP5) was measured to assess OB and OC function, respectively. Interestingly, despite the scaffold’s structured character supporting the growth of the Saos-2 OBs and THP-1-derived OCs coculture, the incorporation of HANPs did not significantly enhance cellular responses compared to scaffolds without HANPs, except for collagen type I production. These findings suggest the need for further investigation into the potential benefits of HANPs in bone tissue engineering applications. Nevertheless, our study contributes valuable insights into optimizing biomaterial design for bone tissue regeneration, with implications for drug screening and material testing protocols. Full article

Background: Oral squamous cell carcinoma (OSCC) frequently invades the jawbone, leading to diagnostic and therapeutic challenges. While tumor–bone interactions have been studied, the specific roles of dental follicle cells (DFCs) and periodontal ligament cells (PDLCs) in OSCC-associated bone resorption remain unclear. This study aimed to compare the effects of DFCs and PDLCs on OSCC-induced bone invasion and elucidate the underlying mechanisms. Methods: Primary human DFCs and PDLCs were isolated from extracted third molars and characterized by Giemsa and immunofluorescence staining. An in vitro co-culture system and an in vivo xenograft mouse model were established using the HSC-2 OSCC cell line. Tumor invasion and osteoclast activation were assessed by hematoxylin and eosin (HE) and tartrate-resistant acid phosphatase (TRAP) staining. Immunohistochemical analysis was performed to evaluate the expression of receptor activator of NF-κB ligand (RANKL) and parathyroid hormone-related peptide (PTHrP). Results: DFCs significantly enhanced OSCC-induced bone resorption by promoting osteoclastogenesis and upregulating RANKL and PTHrP expression. In contrast, PDLCs suppressed RANKL expression and partially modulated PTHrP levels, thereby reducing osteoclast activity. Conclusions: DFCs and PDLCs exert opposite regulatory effects on OSCC-associated bone destruction. These findings underscore the importance of stromal heterogeneity and highlight the therapeutic potential of targeting specific stromal–tumor interactions to mitigate bone-invasive OSCC. Full article

Chinese herbal medicines have played a significant role in the development of new and effective drugs, but how to identify the active ingredients from complex extracts of traditional Chinese herbal medicines was a research difficulty. In recent years, few studies have focused on high-efficiency identification of small-molecule inhibitors of Programmed Death Ligand 1 with lower antigenicity and flexible structure tunability. In order to identify small molecule inhibitors of PD-L1 from complex Chinese herbal extracts, this study established a protein–ligand trapping system based on high-performance liquid chromatography coupled with a photo-diode array detector, ion trap/quadrupole time-of-flight tandem mass spectrometry, and a Programmed Death Ligand 1 affinity chromatography unit (ACPD-L1-HPLC-PDA-IT-TOF (Q-TOF)-MS) to rapidly screen and identify small-molecule inhibitors of Programmed Death Ligand 1 from Toddalia asiatica (L.) Lam. Fourteen components were then identified as PD-L1 binders, and surface plasmon resonance (SPR) validation results showed that six of them—magnoflorine (6), nitidine (22), chelerythrine (24), jatrorrhizine (13), toddaculin (68), and toddanol (45)—displayed PD-L1 binding activity. Laser scanning confocal microscopy results demonstrated that these compounds effectively inhibited the binding of PD-1 to PD-L1 in a dose-dependent manner. Additionally, flow cytometry analysis indicated they could promote human lung cancer cell line (A549) apoptosis when co-cultured with Peripheral Blood Mononuclear Cells (PBMCs). The system’s innovation lies in its first integration of dynamic protein–ligand trapping with multi-dimensional validation, coupled with high-throughput screening capacity for structurally diverse natural products. This workflow overcomes traditional phytochemical screening bottlenecks by preserving native protein conformations during affinity capture while maintaining chromatographic resolution, offering a transformative template for accelerating natural product-derived immunotherapeutics through the PD-1/PD-L1 pathway. Full article

A form of AI was developed and trained to classify four different cell types, with a particular focus on identifying, counting, and determining the maturity of osteoclasts. Osteoclasts, formed by the fusion of monocytes, show clear morphological differences in their maturation, from small mononuclear cells to large multinuclear cells. The developed AI used YOLOv5m models to analyze these cell types based on microscopic images. The AI showed a certain degree of correlation with biochemical analyses (TRAP 5b, CAII). Despite this success, several challenges were identified. The homogeneity of the training data, limited by standardized cell culture conditions, limited the coverage of all osteoclast properties. Furthermore, the AI did not take into account the number of cell nuclei or the specific amount of DNA in the cells, which impaired the precision of the analysis of multinucleated osteoclasts. In the future, the introduction of weighting factors for cell nuclei could optimize the agreement of AI results with biochemical analyses. In summary, the developed AI technology offers a promising tool for cell identification and analysis, especially in osteoclast research. With further developments, this technology could significantly increase the efficiency and accuracy of cell analysis and promote practical applications in research and diagnostics. Full article

Osteoclast differentiation inhibition is a viable treatment strategy for osteoporosis because osteoclasts play a vital role in disease progression. Rhusflavone (Rhus), a biflavonoid, exhibits a sedative–hypnotic effect via the positive allosteric modulation of GABA(A) receptors. Although several biflavonoids possess activities that help prevent bone loss, the potential effects of Rhus on osteoclastogenesis have not been reported yet. In this study, we investigated the effects and underlying biological mechanisms of Rhus isolated from the dried roots of Rhus succedanea on osteoclastogenesis in primary cultured bone marrow-derived macrophages. No cytotoxicity was observed in bone marrow macrophages (BMMs) or during osteoclast differentiation. However, Rhus reduced the number of tartrate-resistant acid phosphatase (TRAP)-positive multinuclear osteoclasts during receptor activator of nuclear factor kappa B ligand (RANKL)-induced osteoclastogenesis. The results of F-actin ring formation demonstrated that Rhus suppresses the bone resorption activity of osteoclasts. Additionally, Rhus inhibits the expression of osteoclast differentiation marker proteins, specifically c-Fos and NF-ATc1. Western blot analysis revealed that Rhus primarily attenuated RANKL-mediated key signaling pathways, particularly the AKT signaling pathway. Furthermore, we found that the AKT activator and inhibitor pharmacologically abolished and enhanced the inhibitory effects of Rhus on osteoclast differentiation, respectively. Taken together, our findings provide evidence that Rhus is a promising biologically active compound that regulates osteoclast differentiation by inhibiting the AKT signaling pathway, which may contribute to future drug development. Full article

Western flower thrips (WFT), Frankliniella occidentalis (Pergande) [Thysanoptera: Thripidae], is a destructive pest of greenhouse ornamentals. Flowering yellow marigolds, Tagetes patula (L.) [Asterales: Asteraceae], have been shown to be attractive to WFT, implicating their suitability as a trap plant. However, functionality may vary in part due to crop variety, the growth stage of the crop in which the marigold trap plant is deployed and whether or not the crop plants have flowers present. The attractiveness of yellow marigolds was tested within several varieties of mature, flowering ornamental bedding plants that were flowering or had their flowers removed: Calibrachoa spp. Petunia spp. Verbena spp., Osteospermum spp., Impatiens haekeri and other marigolds. Trials were conducted in cages under controlled greenhouse conditions for six weeks. The effectiveness of orange vs. yellow marigolds as a trap plant was also assessed. The results showed that WFT were attracted to flowering marigolds and, over time, were present in greater numbers on these than crop plants for all crop species and varieties tested at varying levels of significance. Yellow marigold trap plants were the least effective when deployed within other marigold varieties. In crops with flowers, it took up to five weeks for the number of WFT on trap plants to surpass numbers on crop plants. In contrast, in crop varieties with their flowers removed, trap plants attracted WFT earlier, within one to two weeks, and harbored them in greater numbers over time. Orange, flowering marigolds were a less effective trap plant compared to yellow marigolds. These results confirm that flowering yellow marigolds are attractive to WFT and have potential as a trap plant in greenhouse ornamentals, particularly when crop plants do not have flowers. Full article