Fighting Methicillin-Resistant Staphylococcus aureus with Targeted Nanoparticles.

Antimicrobial resistance (AMR) is considered one of the greatest threats to global health. Methicillin-resistant Staphylococcus aureus (MRSA) remains at the core of this threat, accounting for about 90% of S. aureus infections widespread in the community and hospital settings. In recent years, the use of nanoparticles (NPs) has emerged as a promising strategy to treat MRSA infections. NPs can act directly as antibacterial agents via antibiotic-independent activity and/or serve as drug delivery systems (DDSs), releasing loaded antibiotics. Nonetheless, directing NPs to the infection site is fundamental for effective MRSA treatment so that highly concentrated therapeutic agents are delivered to the infection site while directly reducing the toxicity to healthy human cells. This leads to decreased AMR emergence and less disturbance of the individual's healthy microbiota. Hence, this review compiles and discusses the scientific evidence related to targeted NPs developed for MRSA treatment.

Structural characterization of functionalized gold nanoparticles for drug delivery in cancer therapy: a NMR based approach.

In the present paper, we report results from a study of the structure and physicochemical properties of gold nanoparticles modified with poly(ethylene glycol) (PEG) designed for the drug delivery of the proteasome inhibitor Bortezomib (BTZ) in cancer therapy. A number of advanced analytical techniques were used to define important physicochemical characteristics such as composition, structure, surface properties, particle size and morphology. A new approach based on detailed NMR studies was employed to define specific intermolecular interactions and mechanisms of drug immobilization and location into surface modified gold nanoparticles (AuNPs). Particularly important information was gained from analysis of NMR spectroscopic parameters such as the spectral line shape, translation diffusion, the nuclear Overhauser effect (NOE) and spin-lattice relaxation (T1). The results confirmed the coexistence of two different types of BTZ inclusion into polyethylene glycol coated gold nanoparticles: (i) association with the polymer chains by weak H-bonds and/or dipole-charge interactions and (ii) adsorption on the surface of the gold nanoparticles. The results allowed for determination of the overall structure of Bortezomib loaded PEG coated AuNPs, which is related to the therapeutic drug efficacy and activity in the treatment of cancer.

Graduated compression sleeves: effects on metabolic removal and neuromuscular performance.

The aim of this study was to examine the effects of upper-body graduated compression sleeves (CS) on neuromuscular and metabolic responses during a power training. Fifteen resistance trained men (age: 23.07 ± 3.92 years; body mass: 76.13 ± 7.62 kg; height: 177 ± 6 cm) performed 2 separate power training protocols, either wearing CS or placebo sleeves (PS), in a counterbalanced fashion. Participants first performed a familiarization session and a bench press 1 repetition maximum (1RM) test. The training protocol consisted of 6 sets of 6 repetitions of bench press with a load of 50% 1RM. Statistical analysis compared mean power, peak power, blood lactate, muscle activation, isometric strength, and repetitions to failure. Mean and peak power significantly (p ≤ 0.05) decreased with increasing sets. However, there was no significant difference (p > 0.05) on mean and peak power between protocols. Blood lactate clearance was also not significantly different (p > 0.05) between CS and PS. Muscle activation was not different between PRE and POST (p > 0.05) for any of the muscles analyzed. Isometric strength decreased from PRE to POST (p ≤ 0.05) and was not different between CS and PS. Repetitions to failure were not different between protocols (p > 0.05). These results demonstrate no positive performance effects when wearing graduated CS during power exercise in young trained men.

Dissociated Time Course of Muscle Damage Recovery Between Single- and Multi-Joint Exercises in Highly Resistance-Trained Men.

This study compared the time course of elbow flexor muscle recovery after multi- and single-joint exercises in highly resistance-trained men. Sixteen men (24.5 ± 5.5 years) performed, in a counterbalanced order, 8 sets of 10 repetition maximum (RM) unilateral seated row exercise and 8 sets of 10RM unilateral biceps preacher curl exercise using the contralateral arm. Maximum isometric peak torque (PT) and delayed onset muscle soreness (DOMS) were recorded at baseline (pre), 10 minutes, 24, 48, 72, and 96 hours after each exercise protocol. There was a significant decrease (p ≤ 0.05) in elbow flexor PT 10 minutes after both the multi- and single-joint exercise sessions. However, PT decrease was greater after single-joint (26.8%) when compared with multi-joint (15.1%) exercise (p ≤ 0.05). In addition, elbow flexor PT was lower (8.4%) than baseline 24 hours after the single-joint exercise (p < 0.01), whereas PT returned to baseline 24 hours after the multi-joint exercise. Compared with baseline, DOMS increased at 24, 48, and 72 hours after single-joint exercise (p ≤ 0.05). However, DOMS returned to baseline levels after 72 hours after multi-joint exercise. In addition, DOMS after single-joint exercise was greater (p ≤ 0.05) than after multi-joint exercise at 24, 48, and 72 hours after exercise. Our data suggest that after a resistance training session, highly resistance-trained men experience dissimilar elbow flexor strength recovery between single-joint and multi-joint exercises. Likewise, elbow flexor DOMS is greater and takes longer to recover after single-joint exercise.

Promising strategies employing nucleic acids as antimicrobial drugs.

Antimicrobial resistance (AMR) is a growing concern because it causes microorganisms to develop resistance to drugs commonly used to treat infections. This results in increased difficulty in treating infections, leading to higher mortality rates and significant economic effects. Investing in new antimicrobial agents is, therefore, necessary to prevent and control AMR. Antimicrobial nucleic acids have arisen as potential key players in novel therapies for AMR infections. They have been designed to serve as antimicrobials and to act as adjuvants to conventional antibiotics or to inhibit virulent mechanisms. This new category of antimicrobial drugs consists of antisense oligonucleotides and oligomers, DNAzymes, and transcription factor decoys, differing in terms of structure, target molecules, and mechanisms of action. They are synthesized using nucleic acid analogs to enhance their resistance to nucleases. Because bacterial envelopes are generally impermeable to oligonucleotides, delivery into the cytoplasm typically requires the assistance of nanocarriers, which can affect their therapeutic potency. Given that numerous factors contribute to the success of these antimicrobial drugs, this review aims to provide a summary of the key advancements in the use of oligonucleotides for treating bacterial infections. Their mechanisms of action and the impact of factors such as nucleic acid design, target sequence, and nanocarriers on the antimicrobial potency are discussed.

Caffeic acid loaded into engineered lipid nanoparticles for Alzheimer's disease therapy.

Alzheimer's disease (AD) is an incurable neurological illness and the leading cause of dementia, characterized by amyloid ß (Aß) fibril deposits. Caffeic acid (CA) has demonstrated potential value for AD therapy due to its anti-amyloidogenic, anti-inflammatory, and antioxidant properties. However, its chemical instability and limited bioavailability limit its therapeutic potential in vivo. Herein, liposomes loading CA were produced by distinct techniques. Taking advantage of the overexpression of transferrin (Tf) receptors in brain endothelial cells, Tf was conjugated to the liposomes' surface to direct the CA-loaded nanoparticles (NPs) to the blood-brain barrier (BBB). The optimized Tf-modified NPs exhibited a mean size of around 140 nm, a polydispersity index lower than 0.2, and a neutral surface charge, being appropriate for drug delivery. The Tf-functionalized liposomes showed suitable encapsulation efficiency and physical stability for at least 2 months. Furthermore, in simulated physiological settings, the NPs ensured the sustained release of CA for 8 days. The anti-amyloidogenic efficacy of the optimized drug delivery system (DDS) was investigated. The data show that CA-loaded Tf-functionalized liposomes are capable of preventing Aß aggregation and fibril formation, and disaggregating mature fibrils. Hence, the proposed brain-targeted DDS may be a potential strategy for preventing and treating AD. Future studies in animal models of AD will be valuable to validate the therapeutic efficacy of the optimized nanosystem.

Therapeutic Potential of Natural Compounds in Neurodegenerative Diseases: Insights from Clinical Trials.

Neurodegenerative diseases are caused by the gradual loss of neurons' function. These neurological illnesses remain incurable, and current medicines only alleviate the symptoms. Given the social and economic burden caused by the rising frequency of neurodegenerative diseases, there is an urgent need for the development of appropriate therapeutics. Natural compounds are gaining popularity as alternatives to synthetic drugs due to their neuroprotective properties and higher biocompatibility. While natural compounds' therapeutic effects for neurodegenerative disease treatment have been investigated in numerous in vitro and in vivo studies, only few have moved to clinical trials. This article provides the first systematic review of the clinical trials evaluating natural compounds' safety and efficacy for the treatment of the five most prevalent neurodegenerative disorders: Alzheimer's disease, Parkinson's disease, multiple sclerosis, amyotrophic lateral sclerosis, and Huntington's disease.

Engineered liposomes to deliver nucleic acid mimics in Escherichia coli.

Antisense oligonucleotides (ASOs) composed of nucleic acid mimics (NAMs) monomers are considered as potential novel therapeutic drugs against bacterial infections. However, bacterial envelopes are generally impermeable to naked oligonucleotides. Herein, liposomes loaded with NAMs-modified oligonucleotides (LipoNAMs) were evaluated to deliver ASOs in Escherichia coli. Specifically, we tested several surface modifications that included methoxyPEG conjugated to different lipid anchors or modification of the PEG distal ends with maleimide groups and antibodies. MethoxyPEG coated LipoNAMs showed low delivery efficiency for most bacteria, but maleimide-functionalized PEG LipoNAMs were able to deliver ASOs to nearly half of the bacterial population. Conjugation of antibodies to maleimide-functionalized PEG LipoNAMs increased 1.3-fold the delivery efficiency, enhancing the selectivity towards E. coli and biocompatibility. This work demonstrated for the first time that the coupling of antibodies to PEGylated liposomes can significantly improve the delivery of ASOs in E. coli, which might bring alternative routes for the treatment of bacterial infections in the future.

Transferrin-Functionalized Liposomes for the Delivery of Gallic Acid: A Therapeutic Approach for Alzheimer's Disease.

Senile plaques composed of amyloid ß (Aß) fibrils are considered the leading cause of Alzheimer's disease (AD). Molecules with the ability to inhibit Aß aggregation and/or promote Aß clearance are thus a promising approach for AD therapy. Our group recently demonstrated that gallic acid (GA) has strong anti-amyloidogenic properties. In this study, stealth liposomes were prepared for the delivery of GA for AD therapy. The liposomes were functionalized with transferrin (Tf) to direct them to the brain, since Tf receptors are overexpressed in the endothelial cells of the blood-brain barrier. GA-loaded Tf-functionalized liposomes showed mean diameters of 130 nm, low polydispersity index values, and neutral zeta potential. Moreover, the produced nanocarriers promoted the sustained release of GA over 5 days and are physically stable for 1 month under storage conditions. Furthermore, GA-loaded Tf-functionalized liposomes showed a strong ability to interact with Aß1-42 monomers, slowing down the Aß monomer-to-oligomer and oligomer-to-fibril transitions and decreasing the number of fibrils formed by 56%. In addition, the NPs disaggregated approximately 30% of preformed Aß fibrils. The presented results suggest that Tf-functionalized liposomes could be a viable platform for the brain delivery of GA for AD therapy. Studies with animal models of AD will be valuable for validating the therapeutic efficacy of this novel liposomal formulation.

Transferrin-functionalized liposomes loaded with vitamin VB12 for Alzheimer's disease therapy.

Despite the efforts of the pharmaceutical and research sectors, Alzheimer's disease (AD) remains incurable, imposing the demand for new effective strategies. Vitamin B12 (VB12) has aroused interest due to its in vitro anti-amyloidogenic properties. However, the high molecular weight and hydrophilicity of VB12 are the main obstacles to its clinical application by hindering its passage through the blood-brain barrier (BBB). In recent years, drug delivery systems (DDSs) capable of transporting molecules across the BBB have gained attention for their effective brain delivery. In this work, VB12-loaded liposomes functionalized with transferrin (Tf) were produced, envisaging the dual-targeting of VB12 to the BBB and neuronal cells, due to the overexpression of Tf receptors in these cells. The produced liposomes presented sizes smaller than 200 nm, with low polydispersity and neutral zeta potential, being suitable for brain delivery. The nanoparticles exhibited an adequate encapsulation efficiency, a sustained release of VB12 for 9 days, and physical stability at storage conditions for up to 2 months. The developed nanosystem was capable of delaying the formation of Aß fibrils and disrupting mature fibrils, highlighting its great potential for the prevention and treatment of AD.

Liposome Delivery of Nucleic Acids in Bacteria: Toward In Vivo Labeling of Human Microbiota.

Development of specific probes to study the in vivo spatial distribution of microorganisms is essential to understand the ecology of human microbiota. Herein, we assess the possibility of using liposomes loaded with fluorescently labeled nucleic acid mimics (LipoNAMs) to image Gram-negative and Gram-positive bacteria. We proved that liposome fusion efficiencies were similar in both Gram-negative and Gram-positive bacteria but that the efficiency was highly dependent on the lipid concentration. Notably, LipoNAMs were significantly more effective for the internalization of oligonucleotides in bacteria than the fixation/permeabilization methods commonly used in vitro. Furthermore, a structural and morphological assessment of the changes on bacteria allowed us to observe that liposomes increased the permeability of the cell envelope especially in Gram-negative bacteria. Considering the delivery efficiency and permeabilization effect, lipid concentrations of approximately 5 mM should be selected to maximize the detection of bacteria without compromising the bacterial cellular structure.

Vitamin B12 Inhibits Aß Fibrillation and Disaggregates Preformed Fibrils in the Presence of Synthetic Neuronal Membranes.

The aggregation of amyloid ß (Aß) peptide with subsequent formation of fibrils which deposit in senile plaques is considered one of the key triggers of Alzheimer's disease (AD). Molecules targeting the inhibition of Aß fibrillation and/or the disruption of Aß fibrils are thus promising approaches for the medical prevention and treatment of AD. However, amyloid formation is a complex process strongly influenced by the cellular environment, such as cell membranes, which may affect the effectiveness of therapeutic molecules. In this study, the effect of the vitamin B12 (VB12) on the formation and disaggregation of Aß1-42 fibrils was investigated in the presence of artificial neuronal membranes mimicked by liposomes. Evidence showed that VB12 slows down the Aß fibrillization and reduces the content of fibrils in aqueous solution. Moreover, the vitamin exhibited a strong ability to disrupt preformed fibrils. However, the presence of lipid vesicles compromised the VB12's antiamyloidogenic properties due to the competitive interaction of the vitamin with the lipid membrane and the Aß peptide. Even so, VB12 was effective in inhibiting the fibril formation and disaggregating fibrils in the lipid membrane environment. Thereby, these results indicate that VB12 could be a promising molecule both for the prevention and cure of AD, thus warranting its study in animal models.

Oleogel-Based Systems for the Delivery of Bioactive Compounds in Foods.

Oleogels are semi-solid materials containing a large fraction of liquid oil entrapped in a network of structuring molecules. In the food industry, these formulations can be used to mimic fats and to deliver bioactive compounds. In the last decade, there has been increasing interest in these structures, not only from a scientific point of view, i.e., studying new molecules, methodologies for gelification, and new structures, but also from a technological point of view, with researchers and companies exploring these structures as a way to overcome certain challenges and/or create new and innovative products. One of the exciting applications of oleogels is the delivery of functional molecules, where the incorporation of oil-soluble functional compounds can be explored not only at the macroscale but also at micro- and nanoscales, resulting in different release behaviors and also different applications. This review presents and discusses the most recent works on the development, production, characterization, and applications of oleogels and other oleogel-based systems to deliver functional molecules in foods.

Influence of traffic emissions on the carcinogenic polycyclic aromatic hydrocarbons in outdoor breathable particles.

Because polycyclic aromatic hydrocarbons (PAHs) have been proven to be toxic, mutagenic, and/or carcinogenic, there is widespread interest in analyzing and evaluating exposure to PAHs in atmospheric environments influenced by different emission sources. Because traffic emissions are one of the biggest sources of fine particles, more information on carcinogenic PAHs associated with fine particles needs to be provided. Aiming to further understand the impact of traffic particulate matter (PM) on human health, this study evaluated the influence of traffic on PM10 (PM with aerodynamic diameter < 10 microm) and PM2.5 (PM with aerodynamic diameter < 2.5 microm), considering their concentrations and compositions in carcinogenic PAHs. Samples were collected at one site influenced by traffic emissions and at one reference site using low-volume samplers. Analysis of PAHs was performed by microwave-assisted extraction combined with liquid chromatography (MAE-LC); 17 PAHs, including 9 carcinogenic ones, were quantified. At the site influenced by traffic emissions, PM10 and PM2.5 concentrations were, respectively, 380 and 390% higher than at the background site. When influenced by traffic emissions, the total concentration of nine carcinogenic compounds (naphthalene, chrysene, benzo(a)anthracene, benzo(b) fluoranthene, benzo(k)fluoranthene, benzo(a)pyrene, dibenzo(a,h)anthracene, indeno(1,2,3-cd)pyrene, and dibenzo(a,l)pyrene) was increased by 2400 and 3000% in PM10 and PM2.5, respectively; these nine carcinogenic compounds represented 68 and 74% of total PAHs (sigma(PAHs)) for PM10 and PM2.5, respectively. All PAHs, including the carcinogenic compounds, were mainly present in fine particles. Considering the strong influence of these fine particles on human health, these conclusions are relevant for the development of strategies to protect public health.

Expression of the sucrose binding protein from soybean: renaturation and stability of the recombinant protein.

The sucrose binding protein (SBP) belongs to the cupin family of proteins and is structurally related to vicilin-like storage proteins. In this investigation, a SBP isoform (GmSBP2/S64) was expressed in E. coli and large amounts of the protein accumulated in the insoluble fraction as inclusion bodies. The renatured protein was studied by circular dichroism (CD), intrinsic fluorescence, and binding of the hydrophobic probes ANS and Bis-ANS. The estimated content of secondary structure of the renatured protein was consistent with that obtained by theoretical modeling with a large predominance of beta-strand structure (42%) over the alpha-helix (9.9%). The fluorescence emission maximum of 303 nm for SBP2 indicated that the fluorescent tryptophan was completely buried within a highly hydrophobic environment. We also measured the equilibrium dissociation constant (K(d)) of sucrose binding by fluorescence titration using the refolded protein. The low sucrose binding affinity (K(d)=2.79+/-0.22 mM) of the renatured protein was similar to that of the native protein purified from soybean seeds. Collectively, these results indicate that the folded structure of the renatured protein was similar to the native SBP protein. As a member of the bicupin family of proteins, which includes highly stable seed storage proteins, SBP2 was fairly stable at high temperatures. Likewise, it remained folded to a similar extent in the presence or absence of 7.6M urea or 6.7 M GdmHCl. The high stability of the renatured protein may be a reminiscent property of SBP from its evolutionary relatedness to the seed storage proteins.

Comparison of upper body strength gains between men and women after 10 weeks of resistance training.

Resistance training (RT) offers benefits to both men and women. However, the studies about the differences between men and women in response to an RT program are not conclusive and few data are available about upper body strength response. The aim of this study was to compare elbow flexor strength gains in men and women after 10 weeks of RT. Forty-four college-aged men (22.63 ± 2.34 years) and forty-seven college-aged women (21.62 ± 2.96 years) participated in the study. The RT program was performed two days a week for 10 weeks. Before and after the training period, peak torque (PT) of the elbow flexors was measured with an isokinetic dynamometer. PT values were higher in men in comparison to women in pre- and post-tests (p < 0.01). Both males and females significantly increased elbow flexor strength (p < 0.05); however, strength changes did not differ between genders after 10 weeks of RT program (11.61 and 11.76% for men and women, respectively; p > 0.05). Effect sizes were 0.57 and 0.56 for men and women, respectively. In conclusion, the present study suggests that men and women have a similar upper body strength response to RT.

Electromyographic analyses of muscle pre-activation induced by single joint exercise.

OBJECTIVE: To investigate whether performing a low-intensity, single-joint exercises for knee extensors was an efficient strategy for increasing the number of motor units recruited in the vastus lateralis muscle during a subsequent multi-joint exercises. METHODS: Nine healthy male participants (23.33+/-3.46 yrs) underwent bouts of exercise in which knee extension and 45 degrees , and leg press exercises were performed in sequence. In the low-intensity bout (R30), 15 unilateral knee extensions were performed, followed by 15 repetitions of the leg presses at 30% and 60% of one maximum repetition load (1-MR), respectively. In the high-intensity bout (R60), the same sequence was performed, but the applied load was 60% of 1-MR for both exercises. A single set of 15 repetitions of the leg press at 60% of 1-MR was performed as a control exercise (CR). The surface electromyographic signals of the vastus lateralis muscle were recorded by means of a linear electrode array. The root mean square (RMS) values were determined for each repetition of the leg press, and linear regressions were calculated from these results. The slopes of the straight lines obtained were then normalized using the linear coefficients of the regression equations and compared using one-way ANOVAs for repeated measures. RESULTS: The slopes observed in the CR were significantly lower than those in the R30 and R60 (p<0.05). CONCLUSIONS: The results indicated that the recruitment of motor units was more effective when a single-joint exercise preceded the multi-joint exercise. Article registered in the Australian New Zealand Clinical Trials Registry (ANZCTR) under the number ACTRN12609000413224.

Serum levels of sTNF-R1, sTNF-R2 and CXCL9 correlate with disease activity in adult type paracoccidioidomycosis.

Paracoccidioidomycosis (PCM) is the most common systemic mycosis in Latin America. A major problem in the management of PCM is to determine the best time to discontinue therapy due to the high relapse rate among patients. Soluble TNF receptors (sTNF-R) levels and chemokines are associated with disease activity in several infectious, inflammatory and autoimmune disorders. The aim of the present work was to evaluate levels of sTNF-R1, sTNF-R2 and chemokines in serum of patients with adult type of PCM, before and after antifungal therapy, and to correlate those levels to disease activity. Concentrations of sTNF-R1, sTNF-R2 and CXCL9 were higher in untreated patients and decreased progressively with treatment. The serum marker with the best accuracy to discriminate PCM cases from controls was sTNF-R2. sTNF-R1 did not drop to control levels before 36 months of treatment. CCL2 and CCL3 levels were low at baseline in PCM patients, raised significantly after 12 months of treatment and diminished thereafter. CCL24 levels were higher after 36 months of antifungal therapy in PCM patients. CCL11 levels were not statistically different from control subjects. sTNF-R1, sTNF-R2 and CXCL9 may be useful as laboratory parameters to assess disease activity in PCM patients.

Arabidopsis and tobacco plants ectopically expressing the soybean antiquitin-like ALDH7 gene display enhanced tolerance to drought, salinity, and oxidative stress.

Despite extensive studies in eukaryotic aldehyde dehydrogenases, functional information about the ALDH7 antiquitin-like proteins is lacking. A soybean antiquitin homologue gene, designated GmTP55, has been isolated which encodes a dehydrogenase motif-containing 55 kDa protein induced by dehydration and salt stress. GmTP55 is closely related to the stress-induced plant antiquitin-like proteins that belong to the ALDH7 family. Transgenic tobacco (Nicotiana tabacum) and Arabidopsis (Arabidopsis thaliana) plants constitutively expressing GmTP55 have been obtained in order to examine the physiological role of this enzyme under a variety of stress conditions. Ectopic expression of GmTP55 in both Arabidopsis and tobacco conferred tolerance to salinity during germination and to water deficit during plant growth. Under salt stress, the germination efficiency of both transgenic tobacco and Arabidopsis seeds was significantly higher than that of their control counterparts. Likewise, under progressive drought, the transgenic tobacco lines apparently kept the shoot turgidity to a normal level, which contrasted with the leaf wilt phenotype of control plants. The transgenic plants also exhibited an enhanced tolerance to H(2)O(2)- and paraquat-induced oxidative stress. Both GmTP55-expressing Arabidopsis and tobacco seeds germinated efficiently in medium supplemented with H(2)O(2), whereas the germination of control seeds was drastically impaired. Similarly, transgenic tobacco leaf discs treated with paraquat displayed a significant reduction in the necrotic lesions as compared with control leaves. These transgenic lines also exhibited a lower concentration of lipid peroxidation-derived reactive aldehydes under oxidative stress. These results suggest that antiquitin may be involved in adaptive responses mediated by a physiologically relevant detoxification pathway in plants.