Developing a novel calcium silver zeolite for caries management.

OBJECTIVE: To develop a novel calcium silver zeolite (Ca-Ag-Zeo) and assess its biocompatibility, physiochemical properties and antimicrobial effects. METHODS: Ca-Ag-Zeo was synthesized using ion-exchange method with calcium chloride, silver nitrate and Zeolite X (Zeo). Silver zeolite X (Ag-Zeo) and Zeo were set as control. The chemical structure, morphology, crystal structure and elemental composition of Ca-Ag-Zeo was characterized by X-ray diffraction spectrum, scanning electron microscopy, transmission electron microscopy and energy dispersive spectroscopy, respectively. Its biocompatibility on the human gingival fibroblasts was assessed by cell counting kit-8 assay. Its physiochemical properties were determined by the released calcium and silver ion using Inductive Coupled Plasma Emission Spectrometry for up to 12 weeks. The antimicrobial properties on Streptococcus mutans, Lactobacillus acidophilus, Lactobacillus casei, and Candida albicans were assessed by minimum bactericidal concentration (MBC) or minimum fungicidal concentration (MFC) assay. RESULTS: Ca-Ag-Zeo with a hexagonal cage structure was synthesized. As for biocompatibility, the half-maximal inhibitory concentration (± SD in mg/mL) of Ca-Ag-Zeo, Ag-Zeo and Zeo in human gingival fibroblasts were 0.52 ± 0.05, 0.15 ± 0.01 and 3.35 ± 0.58, respectively (Zeo > Ca-Ag-Zeo > Ag-Zeo; p < 0.05). As for physiochemical properties, the accumulated ion release (± SD in mg) of Ca-Ag-Zeo, Ag-Zeo and Zeo were 0.011 ± 0.003, 0 and 0 for calcium ion, respectively (Ca-Ag-Zeo > Ag-Zeo, Zeo; p < 0.001), and 0.213 ± 0.032, 0.209 ± 0.019 and 0 for silver ion, respectively (Ca-Ag-Zeo, Ag-Zeo > Zeo; p < 0.001). As for anti-microbial ability, the MBC/MFC (mg/mL) of Ca-Ag-Zeo, Ag-Zeo and Zeo were 32, 16 and > 256 against Streptococcus mutans; 32, 16, > 256 against Lactobacillus acidophilus; 16, 16, and 256 against Lactobacillus casei; 0.25, 0.125; and 2, 1, > 256 against Candida albicans, respectively. CONCLUSION: A novel Ca-Ag-Zeo was developed. It presented better biocompatibility compared to Ag-Zeo. It released calcium and silver ions sustainably, and it could inhibit the growth of common cariogenic microorganisms.

Necrosis and Changes in Pigmentation Following Silver Nitrate Application: A Case Report.

ABSTRACT: Silver nitrate has useful antimicrobial and anti-inflammatory effects. However, there are currently no guidelines in place for its use in cauterization and the management of hemostasis. This lack of guidelines has resulted in different approaches being taken in outpatient and healthcare settings, which can lead to a higher risk of adverse effects. The authors present a case that illustrates a classic but exaggerated adverse effect following silver nitrate application.

In planta synthesis of silver nanoparticles and its effect on adventitious shoot growth and withanolide production in Withania somnifera (L.) Dunal.

Silver (Ag) is a non-essential heavy metal with substantial environmental toxicity but an excellent promotor for plant organogenesis. It is used as an elicitor for secondary metabolite production and for in planta synthesis of metal nanoparticles (MNPs). In the present study, the Ag accumulation and reduction capability of in vitro shoots of Withania somnifera and the toxicity and elicitation effect of Ag on in vitro shoots were explored. In vitro shoot cultures of W. somnifera were treated with different concentrations of silver nitrate for a specific treatment period. Growth index, withaferin A, elemental and electron microscopy analyses were done on silver-treated in vitro shoots of W. somnifera. 1 mM silver nitrate treatment for 12 days period was found to give increased growth index (1.425 ± 0.05c) and withaferin A (2.568 ± 0.08e mg g-1) content. The concentration of bioaccumulated Ag in 1 mM silver nitrate treated in vitro shoot was found to be 50.8 ppm. The presence of nano-Ag was also found in the leaves of 1 mM silver nitrate-treated in vitro shoots. In summary, this is the first report portraying the bioaccumulation and in planta reduction capability of the in vitro shoot system of W. somnifera, which makes it a potential medicinal plant of commercial value for silver contaminated soils.

Neurobehavioral, neurotransmitter and redox modifications in Nauphoeta cinerea under mixed heavy metal (silver and mercury) exposure.

Heavy metals are encountered in nature, and are used in several human endeavors, including in dental fillings. It is well known that the safety of metals depends on their chemical form, as well as the dose and route through which biological systems are exposed to them. Here, we used the Nauphoeta cinerea model to examine the mechanism by which salts of the heavy metals used in dental fillings - silver and mercury - exert their neurotoxicity. Nymphs exposed to heavy metals presented with reduced motor and exploratory abilities as they spent more time immobile, especially in the periphery of a novel object, and covered less distance compared with control nymphs. Exposure to AgNO3 and HgCl2 also exacerbated levels of oxidative stress markers (MDA & ROS) and the neurotransmitter regulators - AChE and MAO, while reducing antioxidant activity markers, both in biochemical (thiol & GST) and RT-qPCR (TRX, GST, SOD, Catalase) examinations, in neural tissues of the cockroach. The observed disruptions in neurolocomotor control, synaptic transmission and redox balance explain how heavy metal salts may predispose organisms to neurological disorders.

Antimicrobial efficiency against fish pathogens on the green synthesized silver nanoparticles.

Fish-borne pathogens such as A. hydrophila and F. aquidurense are the most resistant strains in pisciculture farming. Removing the aforementioned pathogens without antibiotics presents a formidable challenge. To overcome this problem, silver nanoparticles (AgNPs) are synthesized using silver nitrate, water medium, and as an AzadirachtaIndica leaf extract via the green synthesis route. X-ray diffraction (XRD) pattern results authenticate the synthesized material is the face-centered cubic structure of silver. The optical absorption edge of the synthesized product was found at the wavelength of 440 nm from the UV-visible spectra, which is confirmed to relate to the Surface Plasmon Resonance peaks of silver particles. In addition, the optical band gap value of the synthesized Ag sample is measured to be 2.81 eV from the obtained optical absorption spectra. EDX spectrum of the synthesized product also supports confirming the silver particle formation. The FT-IR spectra of the neem extract and silver nanoparticles showed their characteristic functional groups, respectively. The presence of bands between 1000 cm-1 to 500 cm-1 indicates to the formation of silver particles. Spherical particles appeared in the synthesized Ag using Scanning Electron Microscopy (SEM) and Transmission Electron Microscopy (TEM). The particle size of Ag NPs was measured as 40 nm and 62 ± 10 nm by TEM and Dynamic Light Scattering (DLS). The zeta potential was also measured as -12 mV showing the synthesized sample's stable nature. Using the DPPH assay, synthesized AgNPs were taken along with the various concentrations of ascorbic acid (20, 40, 60, 80, and 100 µg/mL) to examine the free radical scavenging activity (RSA). RSA value is higher (84 ± 2 %) for synthesized AgNPs at higher concentration (100 µg/mL) than 21 ± 2 % at low concentration (100 µg/mL). The antimicrobial efficacy of the AgNPs against A. hydrophila and F. aquidurense was performed through the agar diffusion method and its results showed the inhibitory zones of the F.aquidurense and A. hydrophila were measured as 25 ± 3 mm, and 28 ± 4 mm respectively. The synthesized Ag particles showed excellent antimicrobial and antioxidant properties confirmed by antimicrobial and DPPH experiments. It implies that the green synthesized silver nanoparticles could be a good alternative for antibiotics in aquaculture farms. The exposure of low concentrations of silver nanoparticles to zebrafish and brine shrimp does not affect the viability and morphology. The exposure of silver nanoparticles in the fisheries in optimized concentration and time could control the fish-borne pathogens without antibiotics.

Mechanisms of Triton X-100 reducing the Ag+-resistance of Enterococcus faecalis.

To investigate the mechanism of Triton X-100 (TX-100) reducing the Ag+-resistance of Enterococcus faecalis (E. faecalis), and evaluate the antibacterial effect of TX-100 + Ag+ against the induced Ag+-resistant E. faecalis (AREf). The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of AgNO3 against E. faecalis with/without TX-100 were determined to verify the enhanced antibacterial activity. Transmission electron microscopy (TEM) was used to observe the morphological changes of E. faecalis after treatment. The intra- and extracellular concentration of Ag+ in treated E. faecalis was evaluated using inductively coupled plasma mass spectrometer (ICP-MS). The changes in cell membrane potential and integrity of treated E. faecalis were also observed using the flow cytometer. Moreover, AREf was induced through continuous exposure to sub-MIC of Ag+ and the antibacterial effect of TX-100 + Ag+ on AREf was further evaluated. The addition of 0.04% TX-100 showed maximal enhanced antibacterial effect of Ag+ against E. faecalis. The TEM and ICP-MS results demonstrated that TX-100 could facilitate Ag+ to enter E. faecalis through changing the membrane structure and integrity. Flow cytometry further showed the effect of TX-100 on membrane potential and permeability of E. faecalis. In addition, the enhanced antibacterial effect of TX-100 + Ag+ was also confirmed on induced AREf. TX-100 can facilitate Ag+ to enter E. faecalis through disrupting the membrane structure and changing the membrane potential and permeability, thus reducing the Ag+-resistance of E. faecalis and enhancing the antibacterial effect against either normal E. faecalis or induced AREf.

Silver nitrate-assisted photo-crosslinking for enhancing the mechanical properties of an alginate/silk fibroin-based 3D scaffold.

Scaffolds play a pivotal role in tissue engineering and serve as vital biological substitutes, providing structural support for cell adhesion and subsequent tissue development. An ideal scaffold must possess mechanical properties suitable for tissue function and exhibit biodegradability. Although synthetic polymer scaffolds offer high rigidity and elasticity owing to their reactive side groups, which facilitate tailored mechanical and rheological properties, they may lack biological cues and cause persistent side effects during degradation. To address these challenges, natural polymers have garnered attention owing to their inherent bioactivity and biocompatibility. However, natural polymers such as silk fibroin (SF) and tyramine-modified alginate (AT) have limitations, including uncontrolled mechanical properties and weak structural integrity. In this study, we developed a blend of SF and AT as a printable biomaterial for extrusion-based 3D printing. Using photocrosslinkable SF/AT inks facilitated the fabrication of complex scaffolds with high printability, thereby enhancing their structural stability. The incorporation of silver nitrate facilitated the tunability of mechanical and rheological behaviors. SF/AT scaffolds with varying stiffness in the physiologically relevant range for soft tissues (51-246 kPa) exhibited excellent biocompatibility, indicating their promising potential for diverse applications in tissue engineering.

Whole-Mount Histochemical Protocols in Zebrafish for a Hands-On Developmental Biology Practical Course in Ecuador.

Research-based education at the undergraduate level is ideal for fostering the training of future scientists. In an undergraduate Developmental Biology course, this learning strategy requires the availability of model species and enough research reagents, not only for technique training but also for the development of student original projects. This might be challenging in most countries, where resources are limited. Hence, there is a need to develop low-cost solutions for use in the classroom. In this study, we describe the optimization and use of two low-cost protocols in zebrafish embryos for hands-on practical sessions and project-based learning in a Developmental Biology undergraduate course in Ecuador. These protocols were designed for the practical and experimental learning of vertebrate meroblastic cleavage, gastrulation, and neural crest differentiation. The proposed protocols have been previously described in the literature and use silver nitrate and alcian blue, two relatively inexpensive reagents, to label cell membranes and cartilage. The silver nitrate protocol allows the study of cell contact formation during cleavage and the identification of cellular changes during gastrulation, including yolk internalization and epiboly. The alcian blue staining allows the analysis of cranial mesenchymal differentiation into cartilage. These protocols are ideal for practical sessions due to their ease of application, quick results, adaptability to the class schedule, and robustness in the hands of beginning researchers. Finally, these protocols are adaptable for research-based class projects.

Purification of triterpenoid saponins and 25R/25S-inokosterone from Achyranthes bidentata Bl. by high-speed countercurrent chromatography coupled silver nitrate coordination.

An effective method by high-speed countercurrent chromatography coordinated with silver nitrate for the preparative separation of sterones and triterpenoid saponins from Achyranthes bidentata Blume was developed. Methyl tert-butyl ether/n-butanol/acetonitrile/water (4:2:3:8, v/v/v/v) was selected for 20-hydroxyecdysone (compound 1), chikusetsusaponin IVa methyl ester (compound 4), 2'-glycan-11-keto-pigmented saponin V (compound 5), as well as a pair of isomers of 25S-inokosterone (compound 2) and 25R-inokosterone (compound 3), which were further purified by silver nitrate coordinated high-speed countercurrent chromatography. What is more, dichloromethane/methanol/isopropanol/water (6:6:1:4, v/v/v/v) was applied for calenduloside E (compound 6), 3ß-[(O-ß-d-glucuronopyranosyl)-oxy]-oleana-11,13-dien-28-oic acid (compound 7), zingibroside R1 (compound 8) and chikusetsusaponin IVa (compound 9). Adding Ag+ to the solvent system resulted in unique selectivity for 25R/25S isomers of inokosterone, which increased the complexing capability and stability of Ag+ coordinated 25S-inokosterone, as well as the α value between them. These results were further confirmed by the computational calculation of geometry optimization and frontier molecular orbitals assay. Comprehensive mass spectrometry and nuclear magnetic resonance analysis demonstrated the structures of the obtained compounds.

Interplay between secondary metabolites and plant hormones in silver nitrate-elicited Arabidopsis thaliana plants.

Plants produce a myriad of specialized compounds in response to threats such as pathogens or pests and different abiotic factors. The stress-related induction of specialized metabolites can be mimicked using silver nitrate (AgNO3) as an elicitor, which application in conservation agriculture has gained interest. In Arabidopsis thaliana, AgNO3 triggers the accumulation of indole glucosinolates (IGs) and the phytoalexin camalexin as well as pheylpropanoid-derived defensive metabolites such as coumaroylagmatins and scopoletin through a yet unknown mechanism. In this work, the role of jasmonic (JA) and salicylic acid (SA) signaling in the AgNO3-triggered specialized metabolite production was investigated. To attain this objective, AgNO3, MeJA and SA were applied to A. thaliana lines impaired in JA or SA signaling, or affected in the endogenous levels of IGs and AGs. Metabolomics data indicated that AgNO3 elicitation required an intact JA and SA signaling to elicit the metabolic response, although mutants impaired in hormone signaling retained certain capacity to induce specialized metabolites. In turn, plants overproducing or abolishing IGs production had also an altered hormonal signaling response, both in the accumulation of signaling molecules and the molecular response mechanisms (ORA59, PDF1.2, VSP2 and PR1 gene expression), which pointed out to a crosstalk between defense hormones and specialized metabolites. The present work provides evidence of a crosstalk mechanism between JA and SA underlying AgNO3 defense metabolite elicitation in A. thaliana. In this mechanism, IGs would act as retrograde feedback signals dampening the hormonal response; hence, expanding the signaling molecule concept.

The biochemical and growth-associated traits of basil (Ocimum basilicum L.) affected by silver nanoparticles and silver.

BACKGROUND: The biochemical and growth changes resulting from exposure of basil (Ocimum basilicum L.) seedlings to silver nanoparticles and silver were investigated. Over a two-week period, seedlings were exposed to different concentrations (0, 40, and 80 ppm) of silver nanoparticles and silver. RESULTS: Our findings revealed that at concentrations of 40 and 80 ppm, both silver nanoparticles and silver nitrate led to decreased weight, root and shoot length, as well as chlorophyll a and b content. Conversely, these treatments triggered an increase in key biochemical properties, such as total phenols, carotenoids and anthocyanins, with silver nanoparticles showing a more pronounced effect compared to silver nitrate. Moreover, the levels of malondialdehyde (MDA) and hydrogen peroxide (H2O2) rose proportionally with treatment concentration, with the nanoparticle treatment exhibiting a more substantial increase. Silver content showed a significant upswing in both roots and leaves as treatment concentrations increased. CONCLUSIONS: Application of varying concentrations of silver nanoparticles and silver nitrate on basil plants resulted in reduced growth and lower chlorophyll content, while simultaneously boosting the production of antioxidant compounds. Notably, anthocyanin, carotenoid, and total phenol increased significantly. However, despite this increase in antioxidant activity, the plant remained unable to fully mitigate the oxidative stress induced by silver and silver nanoparticles.

99mTc-AgNPs-ICG: nanoparticle for hybrid image / 99mTc-AgNPs-ICG: nanopartícula para imagem híbrida / 99mTc-AgNPs-ICG: nanopartícula para imagem híbrida

Introduction: Currently nanotechnology has radically changed the diagnosis of many human pathologies. The aim of this work is to obtain silver nanoparticles for hybrid imaging (99mTc-AgNPs-ICG) having potential clinical imaging applications. Materials and methods: We mixed 2 ml of ascorbic acid (1.7x10-4 M), 5 mCi of 99mTcO4- , 2 ml of citric acid (8.0x10-4 M) and 0.5 ml of silver nitrate (2.5x10-3 M). Solution pH was 5, and it was shaken for 20 minutes at 37º C. Afterwards, 2 µL of Indocyanine Green (1.3x10-3 M) was added (99mTc-AgNPs-ICG). Physiochemical properties of the solution were characterized by UV (λ1 = 420 nm, λ2 = 254 nm) and gamma detector. Fluorescence image, particle size and IR spectrum were evaluated. Results: Silver nanoparticles were obtained in aqueous solution a pH of 5. Their pH, color and spectrum were stable for seven days. Furthermore, the principal peak characterized by HPLC, UV and Gamma detector had similar retention times. Its UV spectrum showed an absorption band of 420 nm, which corresponds to the plasmon absorption band of these nanoparticles. The particle size was 46 nm ± 1.5 nm. The IR spectrum showed absorption bands in 3193, 2624, 1596 y 1212 cm-1. Conclusions: We describe for the first time in literature the synthesis of hybrid (radioactive and fluorescent) silver nanoparticles. Their physiochemical properties were characterized, being stable and their labelling was reproducible having potential biomedical applications.

Introducción: actualmente la nanotecnología ha cambiado radicalmente el diagnóstico de muchas patologías humanas. El objetivo de este trabajo es obtener nanopartículas de plata para imagen híbrida (99mTc-AgNPs-ICG) que tengan potenciales aplicaciones clínicas en imagen. Materiales y métodos: se mezclaron 2 ml de ácido ascórbico (1,7x10-4 M), 5 mCi de 99mTcO4- , 2 ml de ácido cítrico (8,0x10-4 M) y 0,5 ml de nitrato de plata (2,5x10-3 M). El pH de la solución fue 5, y se agitó durante 20 minutos a 37º C. A continuación, se añadieron 2 µl de verde de indocianina (1,3x10-3 M) (99mTc-AgNPs-ICG). Las propiedades fisicoquímicas de la solución se caracterizaron mediante UV (λ1 = 420 nm, λ2 = 254 nm) y detector gamma. Se evaluaron la imagen de fluorescencia, el tamaño de las partículas y el espectro IR. Resultados: se obtuvieron nanopartículas de plata en solución acuosa a un pH de 5. Su pH, color y espectro fueron estables durante siete días. Además, el pico principal caracterizado por HPLC, UV y detector Gamma tenía tiempos de retención similares. Su espectro UV mostró una banda de absorción de 420 nm, que corresponde a la banda de absorción plasmónica de estas nanopartículas. El tamaño de las partículas era de 46 nm ± 1,5 nm. El espectro IR mostró bandas de absorción en 3193, 2624, 1596 y 1212 cm-1. Conclusiones: describimos por primera vez en la literatura la síntesis de nanopartículas de plata híbridas (radioctivas y fluorescentes). Se caracterizaron sus propiedades fisicoquímicas, siendo estables y su etiquetado fue reproducible teniendo potenciales aplicaciones biomédicas.

Introdução: atualmente, a nanotecnologia mudou radicalmente o diagnóstico de muitas patologias humanas. O objetivo deste trabalho é obter nanopartículas de prata para imagens híbridas (99mTc-AgNPs-ICG) com possíveis aplicações de imagens clínicas. Materiais e métodos: misturamos 2 ml de ácido ascórbico (1,7x10-4 M), 5 mCi de 99mTcO4- , 2 ml de ácido cítrico (8,0x10-4 M) e 0,5 ml de nitrato de prata (2,5x10-3 M). O pH da solução era 5 e ela foi agitada por 20 minutos a 37º C. Em seguida, foram adicionados 2 µL de indocianina verde (1,3x10-3 M) (99mTc-AgNPs-ICG). As propriedades físico-químicas da solução foram caracterizadas por UV (λ1 = 420 nm, λ2 = 254 nm) e detector gama. A imagem de fluorescência, o tamanho das partículas e o espectro de infravermelho foram avaliados. Resultados: as nanopartículas de prata foram obtidas em solução aquosa com pH de 5. Seu pH, cor e espectro permaneceram estáveis por sete dias. Além disso, o pico principal caracterizado por HPLC, UV e detector gama teve tempos de retenção semelhantes. Seu espectro de UV mostrou uma banda de absorção de 420 nm, que corresponde à banda de absorção plasmônica dessas nanopartículas. O tamanho da partícula foi de 46 nm ± 1,5 nm. O espectro de IV mostrou bandas de absorção em 3193, 2624, 1596 e 1212 cm-1. Conclusões: descrevemos pela primeira vez na literatura a síntese de nanopartículas de prata híbridas (radioativas e fluorescentes). Suas propriedades físico-químicas foram caracterizadas, sendo estáveis e sua rotulagem foi reprodutível, com possíveis aplicações biomédicas.

99mTc-AgNPs-ICG: nanopartícula para imagen híbrida / 99mTc-AgNPs-ICG: nanoparticle for hybrid image / 99mTc-AgNPs-ICG: nanopartícula para imagem híbrida

Introducción: actualmente la nanotecnología ha cambiado radicalmente el diagnóstico de muchas patologías humanas. El objetivo de este trabajo es obtener nanopartículas de plata para imagen híbrida (99mTc-AgNPs-ICG) que tengan potenciales aplicaciones clínicas en imagen. Materiales y métodos: se mezclaron 2 ml de ácido ascórbico (1.7 x10-4 M), 5 mCi de 99mTcO4-, 2 ml de ácido cítrico (8.0 x 10-4 M) y 0.5 ml de nitrato de plata (2.5 x 10-3 M). El pH de la solución fue 5, y se agitó durante 20 minutos a 37º C. A continuación, se añadieron 2 µl de verde de indocianina (1.3 x 10-3 M) (99mTc-AgNPs-ICG). Las propiedades fisicoquímicas de la solución se caracterizaron mediante UV (λ1 = 420 nm, λ2 = 254 nm) y detector gamma. Se evaluaron la imagen de fluorescencia, el tamaño de las partículas y el espectro IR. Resultados: se obtuvieron nanopartículas de plata en solución acuosa a un pH de 5. Su pH, color y espectro fueron estables durante siete días. Además, el pico principal caracterizado por HPLC, UV y detector Gamma tenía tiempos de retención similares. Su espectro UV mostró una banda de absorción de 420 nm, que corresponde a la banda de absorción plasmónica de estas nanopartículas. El tamaño de las partículas era de 46 nm ± 1,5 nm. El espectro IR mostró bandas de absorción en 3193, 2624, 1596 y 1212 cm-1. Conclusiones: describimos por primera vez en la literatura la síntesis de nanopartículas de plata híbridas (radioactivas y fluorescentes). Se caracterizaron sus propiedades fisicoquímicas, siendo estables y su etiquetado fue reproducible teniendo potenciales aplicaciones biomédicas.

Introduction: Currently nanotechnology has radically changed the diagnosis of many human pathologies. The aim of this work is to obtain silver nanoparticles for hybrid imaging (99mTc-AgNPs-ICG) having potential clinical imaging applications. Materials and methods: We mixed 2 ml of ascorbic acid (1.7x10-4 M), 5 mCi of 99mTcO4-, 2 ml of citric acid (8.0 x 10-4 M) and 0.5 ml of silver nitrate (2.5 x 10-3 M). Solution pH was 5, and it was shaken for 20 minutes at 37º C. Afterwards, 2 µL of Indocyanine Green (1.3 x 10-3 M) was added (99mTc-AgNPs-ICG). Physiochemical properties of the solution were characterized by UV (λ1 = 420 nm, λ2 = 254 nm) and gamma detector. Fluorescence image, particle size and IR spectrum were evaluated. Results: Silver nanoparticles were obtained in aqueous solution a pH of 5. Their pH, color and spectrum were stable for seven days. Furthermore, the principal peak characterized by HPLC, UV and Gamma detector had similar retention times. Its UV spectrum showed an absorption band of 420 nm, which corresponds to the plasmon absorption band of these nanoparticles. The particle size was 46 nm ± 1.5 nm. The IR spectrum showed absorption bands in 3193, 2624, 1596 y 1212 cm-1. Conclusions: We describe for the first time in literature the synthesis of hybrid (radioactive and fluorescent) silver nanoparticles. Their physiochemical properties were characterized, being stable and their labelling was reproducible having potential biomedical applications.

Introdução: Atualmente, a nanotecnologia mudou radicalmente o diagnóstico de muitas patologias humanas. O objetivo deste trabalho é obter nanopartículas de prata para imagens híbridas (99mTc-AgNPs-ICG) com possíveis aplicações de imagens clínicas. Materiais e métodos: Misturamos 2 ml de ácido ascórbico (1.7 x 10-4 M), 5 mCi de 99mTcO4-, 2 ml de ácido cítrico (8.0 x 10-4 M) e 0.5 ml de nitrato de prata (2.5 x 10-3 M). O pH da solução era 5 e ela foi agitada por 20 minutos a 37º C. Em seguida, foram adicionados 2 µL de indocianina verde (1,3x10-3 M) (99mTc-AgNPs-ICG). As propriedades físico-químicas da solução foram caracterizadas por UV (λ1 = 420 nm, λ2 = 254 nm) e detector gama. A imagem de fluorescência, o tamanho das partículas e o espectro de infravermelho foram avaliados. Resultados: As nanopartículas de prata foram obtidas em solução aquosa com pH de 5. Seu pH, cor e espectro permaneceram estáveis por sete dias. Além disso, o pico principal caracterizado por HPLC, UV e detector gama teve tempos de retenção semelhantes. Seu espectro de UV mostrou uma banda de absorção de 420 nm, que corresponde à banda de absorção plasmônica dessas nanopartículas. O tamanho da partícula foi de 46 nm ± 1,5 nm. O espectro de IV mostrou bandas de absorção em 3193, 2624, 1596 e 1212 cm-1. Conclusões: Descrevemos pela primeira vez na literatura a síntese de nanopartículas de prata híbridas (radioativas e fluorescentes). Suas propriedades físico-químicas foram caracterizadas, sendo estáveis e sua rotulagem foi reprodutível, com possíveis aplicações biomédicas.

Spent coffee grounds as a suitable alternative to standard soil in ecotoxicological tests.

Eisenia andrei is considered in OECD and ISO guidelines to be a suitable replacement for Eisenia fetida in ecotoxicological assays. This suggests that other alternative materials and methods could also be used in standard procedures for toxicity testing. The guidelines also favor using less time-consuming procedures and reducing costs and other limitations to ecotoxicological assessments. In recent years, spent coffee grounds (SCG) have been used to produce vermicompost and biochar and as an additive to organic fertilizers. In addition, the physicochemical characteristics of SCG indicate that the material is a suitable substrate for earthworms, with the organisms performing as well as in natural soil. In the present study, a battery of ecotoxicological tests was established with unwashed and washed SCG and a natural reference soil (LUFA 2.2). The test substrates were spiked with different concentrations of silver nitrate. Survival and reproduction of the earthworm E. andrei were assessed under different conditions, along with substrate basal respiration (SBR) as a proxy for microbial activity. Seedling emergence and the germination index of Lepidium sativum were also determined, following standard guidelines. Exposure to silver nitrate had similar effects on earthworm survival and reproduction, as the estimated effective concentrations (EC10 and EC50) in unwashed SCG and LUFA 2.2 overlapped. A hormetic effect was observed for SBR in LUFA 2.2 spiked with 12.8 mg/kg but not in unwashed SCG. Both SBR and root development were inhibited by similar concentrations of silver nitrate in washed SCG. The findings indicate that unwashed SCG could potentially be used as a substrate in E. andrei toxicity tests and support the eventual inclusion of this material in the standard guidelines.

Expression of ethylene biosynthetic genes during flower senescence and in response to ethephon and silver nitrate treatments in Osmanthus fragrans.

BACKGROUND: Sweet osmanthus (Osmanthus fragrans) is an ornamental evergreen tree species in China, whose flowers are sensitive to ethylene. The synthesis of ethylene is controlled by key enzymes and restriction enzymes, 1-aminocyclopropane-1-carboxylic acid synthase (ACS) and 1-aminocyclopropane-1-carboxylic acid oxidase (ACO), which are encoded by multigene families. However, the key synthase responsible for ethylene regulation in O. fragrans is still unknown. OBJECTIVE: This study aims to screen the key ethylene synthase genes of sweet osmanthus flowers in response to ethylene regulation. METHODS: In this study, we used the ACO and ACS sequences of Arabidopsis thaliana to search for homologous genes in the O. fragrans petal transcriptome database. These genes were also analyzed bioinformatically. Finally, the expression levels of O. fragrans were compared before and after senescence, as well as after ethephon and silver nitrate treatments. RESULTS: The results showed that there are five ACO genes and one ACS gene in O. fragrans transcriptome database, and the phylogenetic tree revealed that the proteins encoded by these genes had high homology to the ACS and ACO proteins in plants. Sequence alignment shows that the OfACO1-5 proteins have the 2OG-Fe(II) oxygenase domain, while OfACS1 contains seven conserved domains, as well as conserved amino acids in transaminases and glutamate residues related to substrate specificity. Expression analysis revealed that the expression levels of OfACS1 and OfACO1-5 were significantly higher at the early senescence stage compared to the full flowering stage. The transcripts of the OfACS1, OfACO2, and OfACO5 genes were upregulated by treatment with ethephon. However, out of these three genes, only OfACO2 was significantly downregulated by treatment with AgNO3. CONCLUSION: Our study found that OfACO2 is an important synthase gene in response to ethylene regulation in sweet osmanthus, which would provide valuable data for further investigation into the mechanisms of ethylene-induced senescence in sweet osmanthus flowers.

Macrofungal Mediated Biosynthesis of Silver Nanoparticles and Evaluation of Its Antibacterial and Wound-Healing Efficacy.

Recently, the utilization of biological agents in the green synthesis of nanoparticles has been given interest. In this study, silver nanoparticles were synthesized from an aqueous extract of macrofungus (mushroom), namely Phellinus adamantinus, in a dark room using 20 µL of silver nitrate. Biosynthesized silver nanoparticles were confirmed by analyzing them using a UV-Vis (ultraviolet-visible) spectrophotometer. The synthesized silver nanoparticles were optimized at different pH and temperatures with various dosages of AgNO3 (silver nitrate) and fungal extracts. The synthesized AgNPs (silver nanoparticles) were characterized using TEM (transmission electron microscopy) and EDX (energy-dispersive X-ray) analyses, which confirmed the presence of silver nanoparticles. The size of the nanosilver particles was found to be 50 nm with higher stability. The mycosynthesized AgNPs showed effective antibacterial activity against strains of Gram-positive (Staphylococcus aureus and Bacillus subtilis) and Gram-negative (E. coli and Pseudomonas aeruginosa) bacteria. The minimum inhibitory concentration (MIC) was found to be 3.125 µg/mL by MIC assay. The MTT assay (3-[4,5-dimethylthiazol-2-yl] 2,5-diphenyl-2H-tetrazolium bromide) was performed to study cytotoxicity, and reduced cell viability was recorded at 100 µg/mL. Silver-Polygalacturonic acid-Polyvinyl alcohol ((Ag-PGA)-PVA) nanofiber was prepared using the electrospinning method. The in vitro wound scratch assay was demonstrated to study the wound-healing efficacy of the prepared nanofiber. The wound-healing efficacy of the AgNP-incorporated nanofiber was found to be 20% after 24 h. This study will lay a platform to establish a unique route to the development of a novel nanobiomaterial and its application in antibacterial and wound-healing therapy.

Granulomatous dermatitis caused by Exserohilum mcginnisii infection in a cow.

A 5-y-old, Piedmontese cow had a 4-mo history of ongoing development of skin masses. This was the only cow affected in a herd of 20 cows. Up to 12, hairless, red-to-black, raised nodules-to-plaques were distributed along the dorsum and tail head. Biopsies were taken for histopathology and ancillary testing. An ulcerated skin section contained dermal infiltrates of eosinophils, plasma cells, neutrophils, macrophages, lymphocytes, and multinucleate giant cells, and pyogranulomas. Fungal hyphae were seen within the dermis, multinucleate giant cells, and pyogranulomas. In pyogranulomas, fungi were surrounded by a Splendore-Hoeppli reaction. Dematiaceous (pigmented) hyphae were rarely observed with H&E-stained and unstained (cleared and mounted) sections, but stained well with a Fontana-Masson stain. Exserohilum mcginnisii was identified by fungal culture, followed by PCR assay and sequencing. Exserohilum is a dematiaceous fungus that causes disease in humans and rarely in animals. The use of unstained sections and Fontana-Masson stain are important to demonstrate pigment because dematiaceous fungi have little melanin and appear as hyaline hyphae histologically. PCR assay and sequencing aid in the differentiation and classification of fungal species. To our knowledge, E. mcginnisii dermal granulomas have not been reported previously in cattle.

Comparing recurrence between cautery techniques in pediatric epistaxis.

OBJECTIVE: To compare the risk of recurrent epistaxis between children treated with silver nitrate (SN) in the office or electrocautery (EC) in the operating room (OR). METHODS: Patients aged 2-18 diagnosed with epistaxis (ICD R04.0) in 2018 and treated with SN or EC were retrospectively reviewed. Epistaxis laterality, history of nasal trauma, and personal or family history of a bleeding disorder were recorded. Patients with prior cautery or epistaxis secondary to a procedure were excluded. Recurrence was defined as initial encounter after cautery with documented epistaxis. Patients were followed up into 2022 to track onset of recurrence. Time to recurrence between SN and EC was compared with hazard curves with predictors for recurrence analyzed via Cox's proportional hazard regression. RESULTS: Among 291 patients cauterized for epistaxis, 62 % (n = 181) received SN compared to 38 % (n = 110) who underwent EC. There was significantly higher risk of recurrence when treated with SN compared to EC (Hazard ratio 2.45, 95 % CI: 1.57-3.82, P < 0.0001). Median time to recurrence was not statistically different between techniques (6.39 months (SN) (IQR: 2.33, 14.82) vs. 4.11 months (EC) (IQR: 1.18, 20.86), P = 0.4154). Complication rates were low for both groups (1.16 % (SN) vs. 0 % (EC), P > 0.05). CONCLUSION: Among patients with epistaxis, risk of recurrence is significantly higher in those cauterized with SN compared to EC. Time to recurrence is not significantly different between cautery techniques.