The Commonly Used Stabilizers for Phytochemical-Based Nanoparticles: Stabilization Effects, Mechanisms, and Applications.

Phytochemicals, such as resveratrol, curcumin, and quercetin, have many benefits for health, but most of them have a low bioavailability due to their poor water solubility and stability, quick metabolism, and clearance, which restricts the scope of their potential applications. To overcome these issues, different types of nanoparticles (NPs), especially biocompatible and biodegradable NPs, have been developed. NPs can carry phytochemicals and increase their solubility, stability, target specificity, and oral bioavailability. However, NPs are prone to irreversible aggregation, which leads to NP instability and loss of functions. To remedy this shortcoming, stabilizers like polymers and surfactants are incorporated on NPs. Stabilizers not only increase the stability of NPs, but also improve their characteristics. The current review focused on discussing the state of the art in research on synthesizing phytochemical-based NPs and their commonly employed stabilizers. Furthermore, stabilizers in these NPs were also discussed in terms of their applications, effects, and underlying mechanisms. This review aimed to provide more references for developing stabilizers and NPs for future research.

Development of Multifunctional Nanoencapsulated trans-Resveratrol/Chitosan Nutraceutical Edible Coating for Strawberry Preservation.

Phytochemical nanoencapsulation for nutrient delivery and edible coatings for perishable food preservation are two emerging technologies. Leveraging the strong antimicrobial function of phytochemical nutrients, we propose convergent research to integrate the two technologies by embedding phytochemical-encapsulated nanoparticles in an edible coating on fresh fruits to achieve multiple functions. In particular, we report the study of an edible coating on strawberries that is composited of trans-resveratrol (R)-encapsulated nanoparticles (RNPs) embedded in a chitosan (CS) matrix. The biodegradable and biocompatible RNPs significantly increased the aqueous solubility of R by 150-fold and bioavailability by 3.5-fold after oral administration. Our results demonstrated the abilities of the RNP-embedded CS edible coating to diminish dehydration, prevent nutrient loss, inhibit microbe growth, increase nutraceutical value, preserve strawberry quality, and extend shelf life during storage at both 22 and 4 °C. Such a phytochemical nanoencapsulation-based edible coating is promising for the dual purposes of enhancing nutrient delivery and preserving perishable foods.

Imaging of surface microdomains on individual extracellular vesicles in 3-D.

Extracellular vesicles (EVs) are secreted from all cell types and are intimately involved in tissue homeostasis. They are being explored as vaccine and gene therapy platforms, as well as potential biomarkers. As their size is below the diffraction limit of light microscopy, direct visualizations have been daunting and single-particle studies under physiological conditions have been hampered. Here, direct stochastic optical reconstruction microscopy (dSTORM) was employed to visualize EVs in three-dimensions and to localize molecule clusters such as the tetraspanins CD81 and CD9 on the surface of individual EVs. These studies demonstrate the existence of membrane microdomains on EVs. These were confirmed by Cryo-EM. Individual particle visualization provided insights into the heterogeneity, structure, and complexity of EVs not previously appreciated.

Transcriptional signatures of Zika virus infection in astrocytes.

Astrocytes are an early and important target of Zika virus (ZIKV) infection in the developing brain, but the impacts of infection on astrocyte function remain controversial. Given that nonhuman primate (NHP) models of ZIKV infection replicate aspects of neurologic disease seen in human infections, we cultured primary astrocytes from the brain tissue of infant rhesus macaques and then infected the cells with Asian or African lineage ZIKV to identify transcriptional patterns associated with infection in these cells. The African lineage virus appeared to have greater infectivity and promote stronger antiviral signaling, but infection by either strain ultimately produced typical virus response patterns. Both viruses induced hypoxic stress, but the Asian lineage strain additionally had an effect on metabolic and lipid biosynthesis pathways. Together, these findings describe an NHP astrocyte model that may be used to assess transcriptional signatures following ZIKV infection.

CRISPR based editing of SIV proviral DNA in ART treated non-human primates.

Elimination of HIV DNA from infected individuals remains a challenge in medicine. Here, we demonstrate that intravenous inoculation of SIV-infected macaques, a well-accepted non-human primate model of HIV infection, with adeno-associated virus 9 (AAV9)-CRISPR/Cas9 gene editing construct designed for eliminating proviral SIV DNA, leads to broad distribution of editing molecules and precise cleavage and removal of fragments of the integrated proviral DNA from the genome of infected blood cells and tissues known to be viral reservoirs including lymph nodes, spleen, bone marrow, and brain among others. Accordingly, AAV9-CRISPR treatment results in a reduction in the percent of proviral DNA in blood and tissues. These proof-of-concept observations offer a promising step toward the elimination of HIV reservoirs in the clinic.

Adverse event following live attenuated chikungunya vaccine in a cynomolgus macaque with pre-existing chronic hydrocephalus.

A cynomolgus macaque (Macaca fascicularis) with a pre-existing, undiagnosed, subclinical but severe cerebral hydrocephalus was enrolled in a study of long-term immunogenicity of the IRES/CHIK vaccine. The animal began showing signs of neurological dysfunction post-vaccination, which progressed and ultimately resulted in euthanasia. The underlying brain abnormality was revealed at necropsy and was subsequently investigated with gross and microscopic examination. This becomes the first reported case of an adverse event following administration of a live attenuated vaccine and suggests the possibility of an increased susceptibility risk of unwanted adverse outcome associated with vaccination in populations with pre-existing conditions such as hydrocephalus.

Current and Future Therapeutic Strategies for Lentiviral Eradication from Macrophage Reservoirs.

Macrophages, one of the most abundant populations of leukocytes in the body, function as the first line of defense against pathogen invaders. Human Immunodeficiency virus 1 (HIV-1) remains to date one of the most extensively studied viral infections. Naturally occurring lentiviruses in domestic and primate species serve as valuable models to investigate lentiviral pathogenesis and novel therapeutics. Better understanding of the role macrophages play in HIV pathogenesis will aid in the advancement towards a cure. Even with current efficacy of first- and second-line Antiretroviral Therapy (ART) guidelines and future efficacy of Long Acting Slow Effective Release-ART (LASER-ART); ART alone does not lead to a cure. The major challenge of HIV eradication is viral latency. Latency Reversal Agents (LRAs) show promise as a possible means to eradicate HIV-1 from the body. It has become evident that complete eradication will need to include combinations of various effective therapeutic strategies such as LASER-ART, LRAs, and gene editing. Review of the current literature indicates the most promising HIV eradication strategy appears to be LASER-ART in conjunction with viral and receptor gene modifications via the CRISPR/Cas9 system. Graphical abstract A multimodal approach to HIV treatment including gene editing, LASER-ART, and latency reversal agents may provide a means to achieve HIV eradication.

AAV-mediated gene delivery attenuates neuroinflammation in feline Sandhoff disease.

Sandhoff disease (SD) is a lysosomal storage disorder characterized by the absence of hydrolytic enzyme ß-N-acetylhexosaminidase (Hex), which results in storage of GM2 ganglioside in neurons and unremitting neurodegeneration. Neuron loss initially affects fine motor skills, but rapidly progresses to loss of all body faculties, a vegetative state, and death by five years of age in humans. A well-established feline model of SD allows characterization of the disease in a large animal model and provides a means to test the safety and efficacy of therapeutic interventions before initiating clinical trials. In this study, we demonstrate a robust central nervous system (CNS) inflammatory response in feline SD, primarily marked by expansion and activation of the microglial cell population. Quantification of major histocompatibility complex II (MHC-II) labeling revealed significant up-regulation throughout the CNS with areas rich in white matter most severely affected. Expression of the leukocyte chemokine macrophage inflammatory protein-1 alpha (MIP-1α) was also up-regulated in the brain. SD cats were treated with intracranial delivery of adeno-associated viral (AAV) vectors expressing feline Hex, with a study endpoint 16weeks post treatment. AAV-mediated gene delivery repressed the expansion and activation of microglia and normalized MHC-II and MIP-1α levels. These data reiterate the profound inflammatory response in SD and show that neuroinflammation is abrogated after AAV-mediated restoration of enzymatic activity.

Sepsis and multi-organ dysfunction associated with postgrooming furunculosis in a dog.

BACKGROUND: Postgrooming furunculosis, as previously described in the dog, is a type of pyoderma that typically responds to routine antimicrobial therapy. Systemic clinical signs are common but are usually mild. ANIMAL: A 3-year-old spayed female great dane was presented with marked dorsal furunculosis of 24 h duration. The clinical signs and blood analyses were consistent with sepsis, systemic inflammatory response syndrome and disseminated intravascular coagulopathy (DIC). METHODS AND RESULTS: Skin culture revealed multi-drug resistant Pseudomonas aeruginosa infection. The dog did not respond to aggressive medical therapy including intravenous antibiotic therapy, fresh frozen plasma and haemodynamic support, and was humanely euthanized. Postmortem findings were consistent with postgrooming furunculosis with associated sepsis and DIC affecting multiple organs including the lungs and brain. CONCLUSIONS AND CLINICAL IMPORTANCE: Postgrooming furunculosis can progress to sepsis and multiple organ dysfunction in the dog and can lead to death.

In Vivo Tracking of Streptococcal Infections of Subcutaneous Origin in a Murine Model.

PURPOSE: Generation of plasmin in vivo by Streptococcus pyogenes is thought to localize the active protease complexes to the pathogen surface to aid in tissue dissemination. Here, we chose to follow cutaneous streptococcal infections by the use of non-invasive bioluminescence imaging to determine if this pathogen can be followed by this approach and the extent of bacterial spread in the absence of canonical plasminogen activation by streptokinase. PROCEDURES: Mice were injected subcutaneously with either bioluminescent strains of streptococci, namely Xen20 and Xen10 or S. pyogenes ALAB49. Bioluminescence imaging was performed daily and results were correlated with microbiological and histological analyses. RESULTS: Comparative analysis of chronologic non-invasive datasets indicated that Xen20 did not disseminate from the initial infection site. Contrary to this, microbiological and histological analyses of Xen20 mice for total bacterial burden indicated sepsis and widespread pathogen involvement. CONCLUSIONS: The use of bioluminescence in microbe-based studies requires genomic and pathologic characterization to correlate imaging results with underlying pathology.