Filtering the NMR Spectra of Complex Mixtures through Polymer-Mediated Paramagnetic Spin Relaxation.

A polymer-mediated paramagnetic spin relaxation (PSR) filter is presented for the selective suppression of signals from polymer-interacting species in the 1D and 2D NMR spectra of mixtures. The combined use of Gd3+ and a polymer with a high transverse relaxation enhancement (R2p , which gives a measure of the Gd3+ -complexing ability) results in the suppression of signals from any polymer-interacting component in mixtures, irrespective of their R2p . By using poly(acrylic acid) (PAA) as a model system, we demonstrate selective filtering of the signals of typical low-R2p species (insensitive to Gd3+ ), such as molecular/polymeric cations and non-ionic polymers, which, through PAA recognition (electrostatic/hydrogen-bonding interactions), become exposed to the paramagnetic effect of Gd3+ , while leaving non-PAA-interacting species unaffected. Typical suppression conditions involve PAA (approximately equimolar amount with respect to the species to be filtered) accompanied by sub-mm concentrations of Gd3+ and T2 -filters ≤100 ms. Overall, by exploiting the PSR principles and the recognition properties of polymers, selective NMR filtrations that are not attainable by diffusion, relaxation, or direct PSR filters, can be achieved.

Tuning Spin-Spin Interactions in Radical Dendrimers.

Two generations of polyphosphorhydrazone (PPH) dendrimers were synthesized and fully functionalized with TEMPO radicals via acrylamido or imino group linkers to evaluate the impact of the linker substitution on the radical-radical interactions. A drastic change in the way that the radicals interacted among them was observed by EPR and CV studies: while radicals in Gn -imino-TEMPO dendrimers presented a strong spin-spin interaction, in the Gn -acrylamido-TEMPO ones they acted mainly as independent radicals. This shows that these interactions could be tuned by the solely substitution of the radical linker, opening the perspective of controlling and modulating the extension of these interactions depending on each application. The chemical properties of the linker strongly influence the spin-spin exchange between pendant radicals.

Predicting PSR filters by transverse relaxation enhancements.

The paramagnetic spin relaxation (PSR) filter allows the suppression of the NMR resonances of individual components in mixtures according to their Gd(3+)-complexing ability. The difficulty in predicting this property hampers, however, the widespread application of this filter. Herein we describe that the PSR filter is dominated by the transverse relaxation enhancement (R(2p)) experienced by nuclei in the presence of Gd(3+), so that R(2p) represents a reliable predictive tool of suppression in the 1D and 2D PSR filter of complex mixtures. The robustness of R(2p) as a predictive tool in PSR filters has been demonstrated at different magnetic fields and for the (1)H, (13)C, COSY, and HMQC filtering of commercial multicomponent compositions, including beverages and drugs.

Stepwise filtering of the internal layers of dendrimers by transverse-relaxation-edited NMR.

The characteristic distribution of transverse relaxation times (T2) within dendrimers (shorter values at the core than the periphery) can be exploited in T2-edited 1D and 2D NMR experiments for the stepwise filtering of internal nuclei according to their topology within the dendritic structure. The resulting filtered spectra, which can be conceived as corresponding to virtual hollow dendrimers, benefit from reduced signal overlap, thus facilitating signal assignment and characterization. The generality of the method as a powerful tool in structural and end-group analysis has been confirmed with various dendritic families and nuclei ((1)H, (13)C, (31)P).

The dynamics of dendrimers by NMR relaxation: interpretation pitfalls.

NMR is a powerful tool to study the dynamics of dendrimers. By analogy to linear polymers, shorter T(1) relaxation times have been traditionally associated to less mobile nuclei and hence, dendrimers described with reduced local motions at either the core or the periphery. Herein we report a NMR relaxation study [(1)H and (13)C T(1), T(2); (13)C{(1)H}NOE; various fields and temperatures] which reveals profound differences between the relaxation behavior of dendrimers and linear polymers. Dendrimers show slower dynamics at internal layers and on increasing generation and may display internal nuclei in the slow motional regime with larger T(1) values than the periphery. In contrast to the relaxation properties of linear polymers, these T(1) increments should not be interpreted as resulting from faster dynamics. Only the recording of T(1) data at various temperatures (alternatively, T(2) or NOE at one temperature) ensures the correct interpretation of dendrimer dynamics.

Measuring the quality of primary care in national health surveys: Lessons from Brazil.

BACKGROUND: South Africa started to lead the cross-culturally validation and use of the Primary Care Assessment Tool (PCAT) in Africa, when Professor Bresick filled a gap, as this continent was until then the only one that had never used it in evaluation of primary health care facilities until 2015. AIM: The authors aim to demonstrate that after the consolidation of Bresick's team to an African version of PCAT, it had been adapted to household survey in Brazil. METHODS: In this letter, authors reflect on how Brazil had adapted PCAT to a national random household survey with Brazilian National Institute of Geography and Statistics (IBGE) - the Brazilian Census Bureau. RESULTS: In the the beginning of 2019, Brazilian Ministry of Health brought back the PCAT as the official national primary health care assessment tool. Brazilian National Institute of Geography and Statistics (IBGE) included a new module (set of questions) in its National Health Survey (PNS-2019) and collected more than 100 000 households interviews in about 40% of the country's municipalities. This module had 25 questions of the Brazilian validated version of the adult reduced PCAT. CONCLUSION: We believe that IBGE innovation with the Ministry of Health can encourage South Africa to establish a similar partnership with its National Institute of Statistics (Statistics South Africa) for the country to establish a baseline for future planning of primary health care, for decision-making based on scientific evidence.

Radical Dendrimers Based on Biocompatible Oligoethylene Glycol Dendrimers as Contrast Agents for MRI.

Finding alternatives to gadolinium (Gd)-based contrast agents (CA) with the same or even better paramagnetic properties is crucial to overcome their established toxicity. Herein we describe the synthesis and characterization of entirely organic metal-free paramagnetic macromolecules based on biocompatible oligoethylene glycol dendrimers fully functionalized with 5 and 20 organic radicals (OEG Gn-PROXYL (n = 0, 1) radical dendrimers) with the aim to be used as magnetic resonance imaging (MRI) contrast agents. Conferring high water solubility on such systems is often a concern, especially in large generation dendrimers. Our approach to overcome such an issue in this study is by synthesizing dendrimers with highly water-soluble branches themselves. In this work, we show that the highly water-soluble OEG Gn-PROXYL (n = 0, 1) radical dendrimers obtained showed properties that convert them in good candidates to be studied as contrast agents for MRI applications like diagnosis and follow-up of infectious diseases, among others. Importantly, with the first generation radical dendrimer, a similar r1 relaxivity value (3.4 mM-1s-1) in comparison to gadolinium-diethylenetriamine pentaacetic acid (Gd-DTPA) used in clinics (3.2 mM-1s-1, r.t. 7T) has been obtained, and it has been shown to not be cytotoxic, avoiding the toxicity risks associated with the unwanted accumulation of Gd in the body.

Fully Water-Soluble Polyphosphorhydrazone-Based Radical Dendrimers Functionalized with Tyr-PROXYL Radicals as Metal-Free MRI T1 Contrast Agents.

The finding of alternative imaging probes to Gadolinium (Gd) and other metal based contrast agents (CA) is crucial to overcome their established toxicity. Herein we describe the synthesis and characterization of an entirely organic metal-free magnetic resonance imaging (MRI) contrast agent based on polyphosphorhydrazone (PPH) dendrimers, fully functionalized with up to 48 organic nitroxide radical units. We propose an innovative synthetic procedure based on the use of an amino acid linker (Tyr) coupled to each dendrimer's branch that permits the anchoring of the radicals and at the same time makes possible the control over their water solubility. We demonstrate that the negatively charged resulting PPH Gn-Tyr-PROXYL (n = 0-3) radical dendrimers are excellent candidates to be used as MRI contrast agents, suited for biomedical applications as they show high water solubility, no aggregation problems, and low cytotoxicity, as well as good stability in highly reducing environments. It is achieved a remarkable r1 relaxivity, ca. four times higher (13 mM-1 s-1) than the gold-standard Gd-DTPA used in clinics. Furthermore, the r1 and r2 relaxivity per unit of radical showed an increase with the increase in generation of dendrimers.

Physiological and biochemical insights into the basal level of resistance of two maize hybrids in response to Fusarium verticillioides infection.

Fusarium stalk rot (FSR), caused by Fusarium verticillioides, is one of the most destructive diseases impacting maize yield worldwide. In this study, net carbon assimilation rate (A), stomatal conductance to water vapor (gs), transpiration rate (E), and internal CO2 concentration (Ci) were evaluated on leaves and the activities of enzymes (chitinase (CHI), ß-1-3-glucanase (GLU), phenylalanine ammonia-lyase (PAL), polyphenoloxidase (PPO), catalase (CAT), ascorbate peroxidase (APX), peroxidase (POX)) as well the concentrations of total soluble phenolics (TSP), lignin-thioglycolic acid (LTGA) derivatives, and malondialdehyde (MDA) were evaluated in the internodes and nodes of plants from maize hybrids moderately resistant (BRS 1035) and susceptible (30F35Y) to FSR. The upward relative lesion length (URLL) and radial fungal colonization (RFC) were 46 and 29% lower for the BRS 1035 hybrid in comparison to 30F35Y hybrid, respectively, at 30 after inoculation (dai). For both hybrids, A, gs, and E values significantly decreased while the Ci values increased on infected leaves compared to noninoculated plants. Inoculated plants from BRS 1035 hybrid showed an increase in A compared to inoculated plants from 30F35Y hybrid, and the increase in Ci values was greater for plants from 30F35Y hybrid at 30 dai compared to plants from BRS 1035 hybrid. The CHI, GLU, PPO, CAT, APX, and POX activities increased for inoculated plants from both hybrids compared to the noninoculated plants. In the internodes region, the increase in the activities of CHI (during the infection process of F. verticillioides) and GLU (at earlier stages of F. verticillioides infection) was more pronounced for plants from BRS 1035 hybrid than for plants from 30F35Y hybrid. In the region of the nodes, activities of CHI (during the infection process of F. verticillioides), PAL (at 20 dai), PPO (at 30 dai), and CAT and POX (both at three dai) were more pronounced for plants from BRS 1035 hybrid than for plants from 30F35Y hybrid. In the internodes region, the lower TSP concentration at 30 dai was linked to a high concentration of LTGA derivatives for inoculated plants from BRS 1035 hybrid compared to inoculated plants from 30F35Y hybrid. Taking together, the results of the present study allowed to conclude that the infection by F. verticillioides triggered physiological and biochemical changes on the stalk of maize plants influencing photosynthesis on leaves. A more robust antioxidative metabolism for reactive oxygen species removal in association with an efficient and strong activity of defense enzymes helped to minimize the cellular damage caused by F. verticillioides infection resulting, therefore, in an increase in maize resistance to FSR.

Fast NMR Screening of Macromolecular Complexes by a Paramagnetic Spin Relaxation Filter.

The paramagnetic spin relaxation filter is described for the rapid NMR screening of intermolecular interactions between ligands and macromolecular anionic receptors with large transverse relaxation enhancements (R 2p). The addition of micromolar concentrations of Gd3+ to the mixture produces the immediate broadening/suppression of the NMR signals of interacting species while leaving unaffected those of noncompetitive binders (one-dimensional and two-dimensional experiments). The method is highly sensitive, unveiling interactions that are too weak to generate changes in chemical shifts or relaxation times. It is operationally very simple and hence, it is amenable to ready implementation by nonspecialists. Examples of application such as detecting the formation of interpolymer complexes, cyclodextrin host-guest interactions, and the screening of DNA ligands are included that demonstrate the reliability and broad applicability of the method.

The blockade of AMPA-kainate and NMDA receptors in the dorsal periaqueductal gray reduces the effects of diazepam withdrawal in rats.

It is well established that the most persistent sign of withdrawal from chronic benzodiazepine use in humans is anxiety. In contrast to other types of drugs of abuse, the emergence of this anxiety does not seem to be linked directly to alterations in the levels of dopamine in the mesolimbic system. Some studies have proposed that fear-like behaviors elicited by benzodiazepine withdrawal could be the result either of alterations in the sensitivity of GABAA receptors or in the neuronal hyperexcitability that results from neuroadaptative responses to chronic treatment, probably mediated by glutamate. The increased fear-like behaviors induced by benzodiazepine withdrawal are similar to the defense reaction displayed by animals exposed to dangerous situations or submitted to electrical or chemical stimulation of the dorsal periaqueductal gray (dPAG), a key structure of the brain aversive system. However, the involvement of the dPAG in drug abuse has been investigated only in the context of the physical effects of drug dependence. Thus, in this study we investigated the effects of injections into the dPAG of the glutamic acid diethyl ester (GDEE) and 2-amino-7-phosphonoheptanoate (AP-7) (AMPA-kainate and NMDA receptors antagonists, respectively) on fear-like behaviors promoted by benzodiazepine withdrawal in rats submitted to aversive events (foot-shocks) immediately before chronic diazepam administration in a conditioning place-preference paradigm, using a light-dark box. Our results showed that inhibition of the glutamatergic neurotransmission in the dPAG reduces the consequence of the diazepam withdrawal in rats, implicating the excitatory amino acids of the dPAG in the modulation of the aversive state induced by benzodiazepine drugs withdrawal.

Complexation models of N-(2-carboxyethyl)chitosans with copper(II) ions.

The copper(II) complex formation equilibria of N-(2-carboxyethyl)chitosans with three different degrees of substitution (DS = 0.42, 0.92, and 1.61) were studied in aqueous solution by pH-potentiometric and UV-spectrophotometric techniques. It was demonstrated that the complexation model of CE-chitosans depends on DS: the [Cu(Glc-NR(2))(2)] complexes are predominant for two lower substituted samples ("bridge model", log beta(12) = 10.06 and 11.6, respectively), whereas the increase of DS leads to formation mainly of the [Cu(Glc-NR(2))] complexes ("pendant model", log beta(11) = 6.41). As a model for copper complexation with a disubstituted residue of CE-chitosan, the complex of N-methyliminodipropionate [CuMidp(H(2)O)].(H(2)O) was synthesized and structurally characterized by XRD. The unit cell consists of two crystallographically nonequivalent Cu atoms having slightly distorted square pyramidal coordination; Midp constitutes the basal plane of the pyramid and acts as a tetradentate NO(3) chelate-bridging ligand by the formation of two six-membered chelate rings (average Cu-O 1.99 A, Cu-N 2.04 A) and a bridge via carbonyl O atom (average Cu-O 1.99 A), an apical position is occupied by a water molecule (average Cu-Ow 2.30 A).