Successful aging and its relationship with mindfulness, self-compassion factors, and subjective well-being in older adults.

OBJECTIVES: Aimed to test an association model for Successful Aging (SA), with mindfulness and self-compassion factors as predictor variables, and components of subjective well-being (SWB) as mediating variables in older adults. METHOD: This cross-sectional, descriptive study was conducted using the Google Forms platform and printed questionnaires, applied in an interview format with older adult participants from the Brazilian states of Bahia and Sergipe. A total of 233 older adults participated, residing in the states of Bahia (34.8%) and Sergipe (65.2%), with a mean age of 69.2 years (SD = 7.33). RESULTS: The multiple linear regression (MLR) results indicated that SA was associated with mindfulness, the positive factor of self-compassion, and positive affect (PA). Subsequently, structural equation modeling (SEM) suggested that both mindfulness and self-compassion, which positively correlated with each other, were significantly and positively associated with PA, while PA was positively and significantly associated with SA. CONCLUSION: Mindfulness and the positive facet of self-compassion, mediated by PA, were indirectly associated with SA, while PA was significantly and more strongly associated with SA. Finally, mindfulness and the positive facet of self-compassion were positively correlated with each other.

A high-throughput screening platform to identify nanocarriers for efficient delivery of RNA-based therapies.

RNA-based therapies are highly selective and powerful regulators of biological functions. Non-viral vectors such as nanoparticles (NPs) are very promising formulations for the delivery of RNA-based therapies but their cell targeting, cell internalization and endolysomal escape capacity is rather limited. Here, we present a methodology that combines high-throughput synthesis of light-triggerable NPs and a high-content imaging screening to identify NPs capable of efficiently delivering different type of RNAs. The NPs were generated using polymers synthesized by Michael type addition reactions and they were designed to: (i) efficiently complex coding (mRNAs) and non-coding (miRNAs and/or lncRNAs) RNA molecules, (ii) allow rapid cell uptake and cytoplasmic release of RNA molecules and (iii) target different cell types based on their composition. Furthermore, light-responsive domains were attached to the polymers by distinctive methods to provide diverse disassembly strategies. The most efficient formulations were identified using cell-based assays and high-content imaging analysis. This strategy allows precise delivery of RNA-based therapies and provides an effective design approach to address critical issues in non-viral gene delivery.

A Light-Triggerable Nanoparticle Library for the Controlled Release of Non-Coding RNAs.

RNA-based therapies offer a wide range of therapeutic interventions including the treatment of skin diseases; however, the strategies to efficiently deliver these biomolecules are still limited due to obstacles related to the cellular uptake and cytoplasmic delivery. Herein, we report the synthesis of a triggerable polymeric nanoparticle (NP) library composed of 160 formulations, presenting physico-chemical diversity and differential responsiveness to light. Six formulations were more efficient (up to 500 %) than commercially available lipofectamine in gene-knockdown activity. These formulations showed differential internalization by skin cells and the endosomal escape was rapid (minutes range). The NPs were effective in the release of siRNA and miRNA. Acute skin wounds treated with the top hit NP complexed with miRNA-150-5p healed faster than wounds treated with scrambled miRNA. Light-activatable NPs offer a new strategy to topically deliver non-coding RNAs.

A near infrared light-triggerable modular formulation for the delivery of small biomolecules.

BACKGROUND: Externally triggered drug delivery systems hold considerable promise for improving the treatment of many diseases, in particular, diseases where the spatial-temporal release of the drug is critical to maximize their biological effect whilst minimizing undesirable, off-target, side effects. RESULTS: Herein, we developed a light-triggerable formulation that takes advantage of host-guest chemistry to complex drugs functionalized with a guest molecule and release it after exposure to near infrared (NIR) light due to the disruption of the non-covalent host-guest interactions. The system is composed by a gold nanorod (AuNR), which generates plasmonic heat after exposure to NIR, a thin layer of hyaluronic acid immobilized to the AuNR upon functionalization with a macrocycle, cucurbit[6]uril (CB[6]), and a drug functionalized with a guest molecule that interacts with the macrocycle. For proof of concept, we have used this formulation for the intracellular release of a derivative of retinoic acid (RA), a molecule known to play a key role in tissue development and homeostasis as well as during cancer treatment. We showed that the formulation was able to conjugate approximately 65 µg of RA derivative per mg of CB[6] @AuNR and released it within a few minutes after exposure to a NIR laser. Importantly, the bioactivity of RA released from the formulation was demonstrated in a reporter cell line expressing luciferase under the control of the RA receptor. CONCLUSIONS: This NIR light-triggered supramolecular-based modular platform holds great promise for theranostic applications.

Cardiac involvement in trypanosomiasis in sheep experimentally infected by Trypanosoma vivax (Ziemman, 1905).

The objective of the present study was to evaluate the clinical signs, electrocardiographic signs and evolution of histopathological lesions in the heart of sheep experimentally infected by Trypanosoma vivax during the acute and chronic phases of infection as well as to investigate the presence of parasitic DNA in the heart using polymerase chain reaction (PCR). Twenty-two male sheep were divided into the following four groups: G1, which consisted of six sheep infected by T. vivax that were evaluated until 20 days post-infection (dpi; acute phase); G2, which consisted of six sheep infected by T. vivax that were evaluated until 90 dpi (chronic phase); and G3 and G4 groups, which each consisted of five uninfected sheep. At the end of the experimental period, electrocardiographic evaluations and necroscopic examinations were performed. Fragments of the heart were collected and stained by Hematoxylin-Eosin and Masson's trichrome, and the fragments were also evaluated by PCR for T. vivax. G2 animals presented clinical signs suggestive of heart failure and electrocardiogram alterations characterized by prolonged P, T and QRS complex durations as well as by a cardiac electrical axis shift to the left and increased heart rate. In these animals, mononuclear multifocal myocarditis and interstitial fibrosis were also observed. PCR revealed positivity for T. vivax in two G1 animals and in all G2 animals. Thus, these findings suggested that T. vivax is responsible for the occurrence of cardiac lesions, which are related to heart failure, electrocardiographic alterations and mortality of the infected animals.

Nanoparticles Conjugated with Photocleavable Linkers for the Intracellular Delivery of Biomolecules.

We report the synthesis and characterization of phototriggerable polymeric nanoparticles (NPs) for the intracellular delivery of small molecules and proteins to modulate cell activity. For that purpose, several photocleavable linkers have been prepared providing diverse functional groups as anchoring points for biomolecules.

Investigation of the binding modes of a positively charged pillar[5]arene: internal and external guest complexation.

The selective binding behavior of a trimethylammonium-derived pillar[5]arene towards different guests in aqueous media and under neutral conditions is reported. Although it is known that this macrocycle has the capability to form complexes with guests, we anticipate that the intrinsic pillar shape of the macrocycle with two positively charged rims should allow a diversity of binding modes. The three guests were selected based on their charge and size. The inclusion binding modes and the affinity of the macrocycle to form host-guest complexes were determined by ITC and NMR techniques. We reveal the ability of a cationic water soluble pillar[5]arene to effectively complex two guest molecules, one in each rim, evidencing the diversity of binding modes. Two different structures for 1 : 1 and three for 1 : 2 complexes are reported showing the pillararene ability for internal/external binding.

A journey from calix[4]arene to calix[6] and calix[8]arene reveals more than a matter of size. Receptor concentration affects the stability and stoichiometric nature of the complexes.

The formation of inclusion complexes between lucigenin (N,N'-dimethyl-9,9'-biacridinium dinitrate) and p-sulfonatocalix[n]arenes (SCn; n = 6, 8) was investigated by fluorescence and NMR spectroscopy. Both SC6 and SC8 were found to form 1 : 1 and 1 : 2 host-guest complexes with lucigenin showing up to 109 M-1 binding affinities. Strong quenching of the lucigenin fluorescence upon complexation was observed. Fluorescence regeneration after competitive binding with other potential guests present in solution was used as an indicator displacement assay to characterize the binding mechanism and affinity of alkaline metal ions (Li+, Na+, K+ and Cs+) with SC6 and SC8. The results demonstrate the formation of 1 : 1 and 1 : 2 calixarene : metal complexes with association constants on the order of 103 M-1 and heteroternary calixarene : lucigenin : metal complexes that predominate at metal cation concentrations above the millimolar range. Owing to the ubiquitous presence of metal cations in SCn solutions as counterions (typically Na+), the detailed description of the complexation of these species is crucial to understand and quantify the host-guest binding properties of these receptors. This work demonstrates that both the thermodynamic stability and the stoichiometric nature of the complexes is dependent on the metal ion concentration and, consequently, on the calixarene concentration.

Lactation curve and milk quality of goats experimentally infected with Trypanosoma vivax.

The present study aimed to evaluate the effects of Trypanosoma vivax infection on the shape of the lactation curve and the milk quality of dairy goats experimentally infected with T. vivax. In total, twenty Saanen goats, aged 26-30 months and the same number of calving (two calvings), were divided into two experimental groups: an infected group, consisting of ten goats intravenously infected with 0.5 ml of blood containing approximately 1.25 × 10(5) trypomastigotes of T. vivax and ten uninfected animals as the control group. Clinical tests and hematocrit, parasitemia, and serum biochemistry evaluations were performed on all of the goats. Milk production was measured daily for 152 days by hand milking the goats and weighing the milk. Every seven days, physiochemical analyses were performed to evaluate the milk. Wood's nonlinear model was used to analyze the lactation curve parameters. The infected goats had high levels of parasitemia and hyperthermia, significantly reduced hematocrit, serum total protein, albumin, and glucose levels and increased cholesterol and urea concentrations. Wood's model indicated that the milk production of goats in the infected group declined sharply over a short period of time and produced a flattened yield curve and significant difference (P < 0.05) in the rate of increase of peak milk production, rate of decrease of milk production after the peak, day of peak milk production, and maximum peak milk production compared with that of the control group. Trypanosomiasis also affected the persistency of lactation, which was significantly reduced in goats in the infected group. In addition, the physico-chemical properties of the milk, including the fat content, defatted dry extracts (DDE) and protein content, decreased significantly (P < 0.05) in the goats in the infected group compared with those in the control group. The T. vivax-infected goats showed reduction in milk production, persistence of lactation, and fat levels, the defatted dry extract (DDE) content, and protein, changing the quality of milk.

Host-guest chemistry of a water-soluble pillar[5]arene: evidence for an ionic-exchange recognition process and different complexation modes.

The complexation of an anionic guest by a cationic water-soluble pillararene is reported. Isothermal titration calorimetry (ITC), (1)H NMR, (1)H and (19)F DOSY, and STD NMR experiments were performed to characterize the complex formed under aqueous neutral conditions. The results of ITC and (1)H NMR analyses showed the inclusion of the guest inside the cavity of the pillar[5]arene, with the binding constant and thermodynamic parameters influenced by the counter ion of the macrocycle. NMR diffusion experiments showed that although a fraction of the counter ions are expelled from the host cavity by exchange with the guest, a complex with both counter ions and the guest inside the pillararene is formed. The results also showed that at higher concentrations of guest in solution, in addition to the inclusion of one guest molecule in the cavity, the pillararene can also form an external complex with a second guest molecule.

Interaction of bolaform surfactants with p-sulfonatocalix[4]arene: the role of two positive charges in the binding.

The inclusion binding manners of bolaform surfactants of type C(n)R6(2+) 2Br(-) with different spacer lengths (n = 6, 12) and terminal headgroup volumes (R = methyl, ethyl) by p-sulfonatocalix[4]arene were studied. The combination of ITC parameters (binding constants and complexation enthalpy and entropy) and NMR chemical shifts and NOE cross-peaks obtained upon complexation allows us to propose different binding modes. The results point out that the spacer length has an influence on the binding stoichiometry. The bolaforms with larger spacer lengths between polar head groups enable the formation of 2:1 complexes in addition to 1:1, while with the shorter spacer they form only 1:1 complexes. On the other hand, the formation of 1:1 complexes is not affected by the headgroup volume or the spacer length of the bolaform. Unexpectedly, a complex binding mode was observed where both positive charged terminal groups of the bolaform are accommodated in the cavity of the calixarene. The inclusion of both terminal groups of the guests can be related to its own structure but also evidence the high flexibility of the calixarene.

Nanoparticle-encapsulated retinoic acid for the modulation of bone marrow hematopoietic stem cell niche.

More effective approaches are needed in the treatment of blood cancers, in particular acute myeloid leukemia (AML), that are able to eliminate resistant leukemia stem cells (LSCs) at the bone marrow (BM), after a chemotherapy session, and then enhance hematopoietic stem cell (HSC) engraftment for the re-establishment of the HSC compartment. Here, we investigate whether light-activatable nanoparticles (NPs) encapsulating all-trans-retinoic acid (RA+NPs) could solve both problems. Our in vitro results show that mouse AML cells transfected with RA+NPs differentiate towards antitumoral M1 macrophages through RIG.1 and OASL gene expression. Our in vivo results further show that mouse AML cells transfected with RA+NPs home at the BM after transplantation in an AML mouse model. The photo-disassembly of the NPs within the grafted cells by a blue laser enables their differentiation towards a macrophage lineage. This macrophage activation seems to have systemic anti-leukemic effect within the BM, with a significant reduction of leukemic cells in all BM compartments, of animals treated with RA+NPs, when compared with animals treated with empty NPs. In a separate group of experiments, we show for the first time that normal HSCs transfected with RA+NPs show superior engraftment at the BM niche than cells without treatment or treated with empty NPs. This is the first time that the activity of RA is tested in terms of long-term hematopoietic reconstitution after transplant using an in situ activation approach without any exogenous priming or genetic conditioning of the transplanted cells. Overall, the approach documented here has the potential to improve consolidation therapy in AML since it allows a dual intervention in the BM niche: to tackle resistant leukemia and improve HSC engraftment at the same time.

A roadmap for model-based bioprocess development.

The bioprocessing industry is undergoing a significant transformation in its approach to quality assurance, shifting from the traditional Quality by Testing (QbT) to Quality by Design (QbD). QbD, a systematic approach to quality in process development, integrates quality into process design and control, guided by regulatory frameworks. This paradigm shift enables increased operational efficiencies, reduced market time, and ensures product consistency. The implementation of QbD is framed around key elements such as defining the Quality Target Product Profile (QTPPs), identifying Critical Quality Attributes (CQAs), developing Design Spaces (DS), establishing Control Strategies (CS), and maintaining continual improvement. The present critical analysis delves into the intricacies of each element, emphasizing their role in ensuring consistent product quality and regulatory compliance. The integration of Industry 4.0 and 5.0 technologies, including Artificial Intelligence (AI), Machine Learning (ML), Internet of Things (IoT), and Digital Twins (DTs), is significantly transforming the bioprocessing industry. These innovations enable real-time data analysis, predictive modelling, and process optimization, which are crucial elements in QbD implementation. Among these, the concept of DTs is notable for its ability to facilitate bi-directional data communication and enable real-time adjustments and therefore optimize processes. DTs, however, face implementation challenges such as system integration, data security, and hardware-software compatibility. These challenges are being addressed through advancements in AI, Virtual Reality/ Augmented Reality (VR/AR), and improved communication technologies. Central to the functioning of DTs is the development and application of various models of differing types - mechanistic, empirical, and hybrid. These models serve as the intellectual backbone of DTs, providing a framework for interpreting and predicting the behaviour of their physical counterparts. The choice and development of these models are vital for the accuracy and efficacy of DTs, enabling them to mirror and predict the real-time dynamics of bioprocessing systems. Complementing these models, advancements in data collection technologies, such as free-floating wireless sensors and spectroscopic sensors, enhance the monitoring and control capabilities of DTs, providing a more comprehensive and nuanced understanding of the bioprocessing environment. This review offers a critical analysis of the prevailing trends in model-based bioprocessing development within the sector.

Using calixarenes to model polyelectrolyte surfactant nucleation sites.

The formation of mixed micelles composed of dodecyltrimethylammonium bromide (C(12)TAB) and a hexamethylated p-sulfonatocalix[6]arene (SC6HM) was studied by several techniques. It was found that above the critical aggregation concentration the concentrations of free and micellized surfactant are strongly related to that of SC6HM. When there is free SC6HM in solution, the addition of C(12)TAB mainly results in an increase in the concentration of micellized surfactant, but when all SC6HM has been aggregated, the addition of C(12)TAB results in a substantial increase in the concentration of free surfactant in solution. When the concentration of free surfactant is equal to the critical micelle concentration of the pure system, a second independent aggregation process is observed. This aggregation behavior has many features that are similar to those of more complex systems that involve surfactants in the presence of oppositely charged polyelectrolytes. In this way, calixarenes can serve as simple models to mimic polyelectrolytes and to gain insight into the complex behavior displayed by these macromolecules.

The "true" affinities of metal cations to p-sulfonatocalix[4]arene: a thermodynamic study at neutral pH reveals a pitfall due to salt effects in microcalorimetry.

A microcalorimetric study on the inclusion of monovalent and divalent metal cations by p-sulfonatocalix[4]arene was performed. The thermodynamic parameters for the complexation of alkali metal cations and Ag(+) were obtained for the first time at neutral pH. The Na(+) cation is routinely present as counterion of the calixarene in neutral aqueous solution, and this must be taken into account in the determination of the thermodynamic parameters for the complexation of Na(+) and the other cations by considering a sequential or a competitive binding scheme. The ΔH° and ΔS° values show that the inclusion process is entropically driven, although an influence of the temperature on the complexation reaction indicates that the enthalpic term is also an important contributor. The results also reveal that enthalpy/entropy compensation balances the gain in one contribution against a corresponding loss in the other. The obtained thermodynamic data are in contrast to the results from previous microcalorimetric studies, which showed binding constants that were orders of magnitude smaller and complexations, which were in part enthalpically driven but which neglected the influence of the alkali metal counterions.

Aggregation of p-Sulfonatocalixarene-Based Amphiphiles and Supra-Amphiphiles.

p-Sulfonatocalixarenes are a special class of water soluble macrocyclic molecules made of 4-hydroxybenzenesulfonate units linked by methylene bridges. One of the main features of these compounds relies on their ability to form inclusion complexes with cationic and neutral species. This feature, together with their water solubility and apparent biological compatibility, had enabled them to emerge as one the most important host receptors in supramolecular chemistry. Attachment of hydrophobic alkyl chains to these compounds leads to the formation of macrocyclic host molecules with amphiphilic properties. Like other oligomeric surfactants, these compounds present improved performance with respect to their monomeric counterparts. In addition, they hold their recognition abilities and present several structural features that depend on the size of the macrocycle and on the length of the alkyl chain, such as preorganization, flexibility and adopted conformations, which make these molecules very interesting to study structure-aggregation relationships. Moreover, the recognition abilities of p-sulfonatocalixarenes enable them to be applied in the design of amphiphiles constructed from non-covalent, rather than covalent, bonds (supramolecular amphiphiles). In this review, we summarize the developments made on the design and synthesis of p-sulfonatocalixarenes-based surfactants, the characterization of their self-assembly properties and on how their structure affects these properties.

A Polymeric Nanoparticle Formulation for Targeted mRNA Delivery to Fibroblasts.

Messenger RNA (mRNA)-based therapies offer enhanced control over the production of therapeutic proteins for many diseases. Their clinical implementation warrants formulations capable of delivering them safely and effectively to target sites. Owing to their chemical versatility, polymeric nanoparticles can be designed by combinatorial synthesis of different ionizable, cationic, and aromatic moieties to modulate cell targeting, using inexpensive formulation steps. Herein, 152 formulations are evaluated by high-throughput screening using a reporter fibroblast model sensitive to functional delivery of mRNA encoding Cre recombinase. Using in vitro and in vivo models, a polymeric nanoformulation based on the combination of 3 specific monomers is identified to transfect fibroblasts much more effectively than other cell types populating the skin, with superior performance than lipid-based transfection agents in the delivery of Cas9 mRNA and guide RNA. This tropism can be explained by receptor-mediated endocytosis, involving CD26 and FAP, which are overexpressed in profibrotic fibroblasts. Structure-activity analysis reveals that efficient mRNA delivery required the combination of high buffering capacity and low mRNA binding affinity for rapid release upon endosomal escape. These results highlight the use of high-throughput screening to rapidly identify chemical features towards the design of highly efficient mRNA delivery systems targeting fibrotic diseases.

Enhanced production of N-acetyl-glucosaminidase by marine Aeromonas caviae CHZ306 in bioreactor.

N-Acetyl-glucosaminidases (GlcNAcases) are exoenzymes found in a wide range of living organisms, which have gained great attention in the treatment of disorders related to diabetes, Alzheimer's, Tay-Sachs', and Sandhoff's diseases; the control of phytopathogens; and the synthesis of bioactive GlcNAc-containing products. Aiming at future industrial applications, in this study, GlcNAcase production by marine Aeromonas caviae CHZ306 was enhanced first in shake flasks in terms of medium composition and then in bench-scale stirred-tank bioreactor in terms of physicochemical conditions. Stoichiometric balance between the bioavailability of carbon and nitrogen in the formulated culture medium, as well as the use of additional carbon and nitrogen sources, played a central role in improving the bioprocess, considerably increasing the enzyme productivity. The optimal cultivation medium was composed of colloidal α-chitin, corn steep liquor, peptone A, and mineral salts, in a 5.2 C:N ratio. Optimization of pH, temperature, colloidal α-chitin concentration, and kLa conditions further increased GlcNAcase productivity. Under optimized conditions in bioreactor (i.e., 34 °C, pH 8 and kLa 55.2 h-1), GlcNAcase activity achieved 173.4 U.L-1 after 12 h of cultivation, and productivity no less than 14.45 U.L-1.h-1 corresponding to a 370-fold enhancement compared to basal conditions.

Independent pathway formation of guest-host in host ternary complexes made of ammonium salt, calixarene, and cyclodextrin.

The interaction between γ-cyclodextrin and amphiphilic p-sulfonatocalix[4]arenes was studied using NMR and isothermal titration calorimetry techniques. The results indicate that these calixarenes are able to form 1:1, 1:2, and 2:1 host-guest complexes with the cyclodextrin. The ROESY spectra suggest that the cyclodextrin binds the calixarenes through the hydrophobic alkyl chains. p-Sulfonatocalix[4]arenes, which are traditionally used as host molecules, act as guests in the presence of γ-cyclodextrin. However, their recognition site remains active upon complexation with the cyclodextrin, and ternary complexes can be devised. Here, we also demonstrate the formation of such complexes using tetramethylammonium chloride as a model guest. Moreover, it is also demonstrated that the recognition properties of the calixarene are unaffected upon complexation with the cyclodextrin.