Clathrin-nanoparticles deliver BDNF to hippocampus and enhance neurogenesis, synaptogenesis and cognition in HIV/neuroAIDS mouse model.

Brain derived neurotrophic factor (BDNF) promotes the growth, differentiation, maintenance and survival of neurons. These attributes make BDNF a potentially powerful therapeutic agent. However, its charge, instability in blood, and poor blood brain barrier (BBB) penetrability have impeded its development. Here, we show that engineered clathrin triskelia (CT) conjugated to BDNF (BDNF-CT) and delivered intranasally increased hippocampal BDNF concentrations 400-fold above that achieved previously with intranasal BDNF alone. We also show that BDNF-CT targeted Tropomyosin receptor kinase B (TrkB) and increased TrkB expression and downstream signaling in iTat mouse brains. Mice were induced to conditionally express neurotoxic HIV Transactivator-of-Transcription (Tat) protein that decreases BDNF. Down-regulation of BDNF is correlated with increased severity of HIV/neuroAIDS. BDNF-CT enhanced neurorestorative effects in the hippocampus including newborn cell proliferation and survival, granule cell neurogenesis, synaptogenesis and increased dendritic integrity. BDNF-CT exerted cognitive-enhancing effects by reducing Tat-induced learning and memory deficits. These results show that CT bionanoparticles efficiently deliver BDNF to the brain, making them potentially powerful tools in regenerative medicine.

Identification and Differential Abundance of Mitochondrial Genome Encoding Small RNAs (mitosRNA) in Breast Muscles of Modern Broilers and Unselected Chicken Breed.

Background: Although small non-coding RNAs are mostly encoded by the nuclear genome, thousands of small non-coding RNAs encoded by the mitochondrial genome, termed as mitosRNAs were recently reported in human, mouse and trout. In this study, we first identified chicken mitosRNAs in breast muscle using small RNA sequencing method and the differential abundance was analyzed between modern pedigree male (PeM) broilers (characterized by rapid growth and large muscle mass) and the foundational Barred Plymouth Rock (BPR) chickens (characterized by slow growth and small muscle mass). Methods: Small RNA sequencing was performed with total RNAs extracted from breast muscles of PeM and BPR (n = 6 per group) using the 1 × 50 bp single end read method of Illumina sequencing. Raw reads were processed by quality assessment, adapter trimming, and alignment to the chicken mitochondrial genome (GenBank Accession: X52392.1) using the NGen program. Further statistical analyses were performed using the JMP Genomics 8. Differentially expressed (DE) mitosRNAs between PeM and BPR were confirmed by quantitative PCR. Results: Totals of 183,416 unique small RNA sequences were identified as potential chicken mitosRNAs. After stringent filtering processes, 117 mitosRNAs showing >100 raw read counts were abundantly produced from all 37 mitochondrial genes (except D-loop region) and the length of mitosRNAs ranged from 22 to 46 nucleotides. Of those, abundance of 44 mitosRNAs were significantly altered in breast muscles of PeM compared to those of BPR: all mitosRNAs were higher in PeM breast except those produced from 16S-rRNA gene. Possibly, the higher mitosRNAs abundance in PeM breast may be due to a higher mitochondrial content compared to BPR. Our data demonstrate that in addition to 37 known mitochondrial genes, the mitochondrial genome also encodes abundant mitosRNAs, that may play an important regulatory role in muscle growth via mitochondrial gene expression control.

A Radish Basic Helix-Loop-Helix Transcription Factor, RsTT8 Acts a Positive Regulator for Anthocyanin Biosynthesis.

The MYB-bHLH-WDR (MBW) complex activates anthocyanin biosynthesis through the transcriptional regulation. RsMYB1 has been identified as a key player in anthocyanin biosynthesis in red radish (Raphanus sativus L.), but its partner bHLH transcription factor (TF) remains to be determined. In this study, we isolated a bHLH TF gene from red radish. Phylogenetic analysis indicated that this gene belongs to the TT8 clade of the IIIF subgroup of bHLH TFs, and we thus designated this gene RsTT8. Subcellular localization analysis showed that RsTT8-sGFP was localized to the nuclei of Arabidopsis thaliana protoplasts harboring the RsTT8-sGFP construct. We evaluated anthocyanin biosynthesis and RsTT8 expression levels in three radish varieties (N, C, and D) that display different red phenotypes in the leaves, root flesh, and root skins. The root flesh of the C variety and the leaves and skins of the D variety exhibit intense red pigmentation; in these tissues, RsTT8 expression showed totally positive association with the expression of RsMYB1 TF and of five of eight tested anthocyanin biosynthesis genes (i.e., RsCHS, RsCHI, RsF3H, RsDFR, and RsANS). Heterologous co-expression of both RsTT8 and RsMYB1 in tobacco leaves dramatically increased the expression of endogenous anthocyanin biosynthesis genes and anthocyanin accumulation. Furthermore, a yeast two-hybrid assay showed that RsTT8 interacts with RsMYB1 at the MYB-interacting region (MIR), and a transient transactivation assay indicated that RsTT8 activates the RsCHS and RsDFR promoters when co-expressed with RsMYB1. Complementation of the Arabidopsis tt8-1 mutant, which lacks red pigmentation in the leaves and seeds, with RsTT8 restored red pigmentation, and resulted in high anthocyanin and proanthocyanidin contents in the leaves and seeds, respectively. Together, these results show that RsTT8 functions as a regulatory partner with RsMYB1 during anthocyanin biosynthesis.

Conditional Human Immunodeficiency Virus Transactivator of Transcription Protein Expression Induces Depression-like Effects and Oxidative Stress.

BACKGROUND: The prevalence of major depression in those with HIV/AIDS is substantially higher than in the general population. Mechanisms underlying this comorbidity are poorly understood. HIV-transactivator of transcription (Tat) protein, produced and excreted by HIV, could be involved. We determined whether conditional Tat protein expression in mice is sufficient to induce depression-like behaviors and oxidative stress. Further, as oxidative stress is associated with depression, we determined whether decreasing or increasing oxidative stress by administering methylsulfonylmethane (MSM) or diethylmaleate (DEM), respectively, altered depression-like behavior. METHODS: GT-tg bigenic mice received intraperitoneal saline or doxycycline (Dox, 25-100 mg/kg/day) to induce Tat expression. G-tg mice, which do not express Tat protein, also received Dox. Depression-like behavior was assessed with the tail suspension test (TST) and the two-bottle saccharin/water consumption task. Reactive oxygen/nitrogen species (ROS/RNS) were assessed ex vivo. Medial frontal cortex (MFC) oxidative stress and temperature were measured in vivo with 9.4-Tesla proton magnetic resonance spectroscopy (MRS). RESULTS: Tat expression increased TST immobility time in an exposure-dependent manner and reduced saccharin consumption. MSM decreased immobility time while DEM increased it in saline-treated GT-tg mice. Tat and MSM behavioral effects persisted for 28 days. Tat and DEM increased while MSM decreased ROS/RNS levels. Tat expression increased MFC glutathione levels and temperature. CONCLUSIONS: Tat expression induced rapid and enduring depression-like behaviors and oxidative stress. Increasing/decreasing oxidative stress increased/decreased, respectively, depression-like behavior. Thus, Tat produced by HIV may contribute to the high depression prevalence among those with HIV. Further, mitigation of oxidative stress could reduce depression severity.

An Integrated Biochemical, Proteomics, and Metabolomics Approach for Supporting Medicinal Value of Panax ginseng Fruits.

Panax ginseng roots are well known for their medicinal properties and have been used in Korean and Chinese traditional medicines for 1000s of years. However, the medicinal value of P. ginseng fruits remain poorly characterized. In this study, we used an integrated biochemical, proteomics, and metabolomics approach to look into the medicinal properties of ginseng fruits. DPPH (1,1-diphenyl-2-picrylhydrazyl) and ABTS [2,2'-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)] assays showed higher antioxidant activities in ginseng fruits than leaves or roots. Two-dimensional gel electrophoresis (2-DE) profiling of ginseng fruit proteins (cv. Cheongsun) showed more than 400 spots wherein a total of 81 protein spots were identified by mass spectrometry using NCBInr, UniRef, and an in-house developed RNAseq (59,251 protein sequences)-based databases. Gene ontology analysis showed that most of the identified proteins were related to the hydrolase (18%), oxidoreductase (16%), and ATP binding (15%) activities. Further, a comparative proteome analysis of four cultivars of ginseng fruits (cvs. Yunpoong, Gumpoong, Chunpoong, and Cheongsun) led to the identification of 22 differentially modulated protein spots. Using gas chromatography-time of flight mass spectrometry (GC-TOF MS), 66 metabolites including amino acids, sugars, organic acids, phenolic acids, phytosterols, tocopherols, and policosanols were identified and quantified. Some of these are well known medicinal compounds and were not previously identified in ginseng. Interestingly, the concentration of almost all metabolites was higher in the Chunpoong and Gumpoong cultivars. Parallel comparison of the four cultivars also revealed higher amounts of the medicinal metabolites in Chunpoong and Gumpoong cultivars. Taken together, our results demonstrate that ginseng fruits are a rich source of medicinal compounds with potential beneficial health effects.

Solid-state NMR structure of a pathogenic fibril of full-length human α-synuclein.

Misfolded α-synuclein amyloid fibrils are the principal components of Lewy bodies and neurites, hallmarks of Parkinson's disease (PD). We present a high-resolution structure of an α-synuclein fibril, in a form that induces robust pathology in primary neuronal culture, determined by solid-state NMR spectroscopy and validated by EM and X-ray fiber diffraction. Over 200 unique long-range distance restraints define a consensus structure with common amyloid features including parallel, in-register ß-sheets and hydrophobic-core residues, and with substantial complexity arising from diverse structural features including an intermolecular salt bridge, a glutamine ladder, close backbone interactions involving small residues, and several steric zippers stabilizing a new orthogonal Greek-key topology. These characteristics contribute to the robust propagation of this fibril form, as supported by the structural similarity of early-onset-PD mutants. The structure provides a framework for understanding the interactions of α-synuclein with other proteins and small molecules, to aid in PD diagnosis and treatment.

A pulse sequence optimization method for assessment of nucleus size in q-space analysis of idealized cells.

To adjust pulse sequences that produce diffusion-weighted MRI signals for increased sensitivity to nucleus size, the impulse-propagator method in q-space is applied to a spherical geometry that would describe each member of a collection of cells and their nuclei, with several possible representations of the extracellular space. The method is extended to allow propagation between nucleus, cytoplasm, and extracellular space through semi-permeable membranes, using an approximate adjustment of intra-compartment propagators. Diffraction patterns are first calculated for the three compartments separately, for PGSE and OGSE pulse sequences, and verified by comparison with Monte Carlo simulations. The detailed patterns from the separate compartments determine the q value for maximum contrast in the total signal between large and small nuclei, an optimization that is not accurate in a Gaussian Phase Distribution (GPD) approximation. Then diffraction patterns are calculated for the case of linked compartments with semi-permeable membranes. The treatment of permeability adequately estimates pulse-sequence parameters for maximum contrast in calculated signal as nucleus size varies.

The effect of acute distal radioulnar joint laxity on outcome after volar plate fixation of distal radius fractures.

OBJECTIVES: The objective of this study was to determine whether intraoperative laxity of the distal radioulnar joint (DRUJ) is associated with adverse postoperative outcomes after volar plate fixation of a distal radius fracture (DRF) and 4 weeks of immobilization. DESIGNS: Prospective study with clinical and radiographic assessment. SETTING: Level 1 trauma center. PATIENTS: One hundred consecutive patients were treated by volar locking plate fixation at our institution for an unstable DRF from April 2007 to November 2009. Of these patients, 84 patients with a minimum follow-up of 12 months were enrolled in this study. INTERVENTION: Intraoperative DRUJ laxity was evaluated using a radioulnar stress test after fixation of DRF using volar locking plate and splint immobilization of the forearm for 1 month in patients with intraoperative DRUJ laxity. Patients were allocated to an unstable group or stable group according to the presence of intraoperative DRUJ laxity. MAIN OUTCOME MEASUREMENTS: Our primary outcome measure was disabilities of arm, shoulder, and hand score and the secondary outcome measures were wrist motion, grip strength, modified Mayo wrist score, visual analogue scale for wrist pain, and ongoing pain in the DRUJ. RESULTS: Nineteen of the 84 study subjects were allocated to the unstable group and 65 to the stable group. No significant differences were observed between 2 groups in wrist range of motion, grip strength, modified Mayo wrist score, disabilities of arm, shoulder, and hand score, visual analogue scale score, and ongoing pain in the DRUJ at 1 year postoperatively. CONCLUSIONS: In our series of patients treated with volar locking plate and immobilization of the forearm for 1 month in patients with intraoperative laxity of the DRUJ, laxity did not affect impairment, pain, or disability 1 year after fracture. However, the role of postoperative immobilization of the forearm is debatable and merits additional study. LEVEL OF EVIDENCE: Prognostic level I.

Adhesive-enhanced sternal closure to improve postoperative functional recovery: a pilot, randomized controlled trial.

BACKGROUND: We previously established a proof-of-concept in a human cadaveric model where conventional wire cerclage was augmented with a novel biocompatible bone adhesive that increased mechanical strength and early bone stability. We report the results of a single-center, pilot, randomized clinical trial of the effects of adhesive-enhanced closure of the sternum on functional postoperative recovery. METHODS: In 55 patients undergoing primary sternotomy, 26 patients underwent conventional wire closure and were compared with 29 patients who underwent adhesive-enhanced closure, which consisted of Kryptonite biocompatible adhesive (Doctors Research Group Inc, Southbury, CT) applied to each sternal edge in addition to conventional 7-wire cerclage. Patients were monitored postoperatively at 72 hours, weekly for 12 weeks, and then after 12 months for incisional pain, analgesic use, and maximal inspiratory capacity measured by spirometry. Standardized assessment tools measured postoperative physical disability and health-related quality of life. RESULTS: No adverse events or sternal complications from the adhesive were observed early or after 12 months. Incisional pain and narcotic analgesic use were reduced in adhesive-enhanced closure patients. Inspiratory capacity was significantly improved, postoperative health-related quality of life scores normalized more rapidly, and physical disability scores were reduced. Computed tomography imaging was suggestive of sternal healing. CONCLUSIONS: Adhesive-enhanced closure is a safe and simple addition to conventional wire closure, with demonstrated benefits on functional recovery, respiratory capacity, incisional pain, and analgesic requirements. A large, multicenter, randomized controlled trial to examine the potential of the adhesive to prevent major sternal complications in higher risk patients is warranted.

Beta-lactam antibiotic decreases acquisition of and motivation to respond for cocaine, but not sweet food, in C57Bl/6 mice.

No medication is approved to treat cocaine addiction, but mounting evidence suggests that glutamate-directed approaches may reduce cocaine dependence and relapse. We tested the hypotheses that the glutamate transporter subtype 1 activator, ceftriaxone, disrupts acquisition of cocaine self-administration, motivation to self-administer cocaine, and conditioned place preference in mice. Repeated ceftriaxone (200 mg/kg) reduced the ability of mice to acquire cocaine and the motivation to self-administer cocaine after successful acquisition without affecting acquisition of or motivation for sweet food. Repeated ceftriaxone had no effect on cocaine-conditioned place preference. These results suggest that a ß-lactam antibiotic reduces the direct reinforcing strength of cocaine without producing nonspecific deficits in conditioned learning processes.

ß-Lactam antibiotic produces a sustained reduction in extracellular glutamate in the nucleus accumbens of rats.

We investigated the short- and long-term effects of ceftriaxone on glutamate transporter subtype 1 (GLT-1) transporter activity and extracellular glutamate in the rat nucleus accumbens. Repeated ceftriaxone administration (50, 100 or 200 mg/kg, i.p.) produced a dose-dependent reduction in glutamate levels that persisted for 20 days following discontinuation of drug exposure. The ceftriaxone effect was prevented by the GLT-1 transporter inhibitor dihydrokainate (1 µM, intra-accumbal). These results suggest that ß-lactam antibiotics produce an enduring reduction in glutamatergic transmission in the brain reward center.

Methanandamide blocks amphetamine-induced behavioral sensitization in rats.

Methanandamide acts at targets which modulate amphetamine-induced behaviors. Therefore, we investigated methanandamide effects on the acute hyperactivity produced by a single injection of amphetamine and behavioral sensitization induced by repeated amphetamine exposure in rats. Methanandamide (5mg/kg, i.p.) did not affect basal locomotor or stereotypical activity. Methanandamide (5mg/kg, i.p.) pretreatment did not alter the acute increase in locomotor or stereotypical activities produced by acute amphetamine (2mg/kg, i.p.). For chronic studies, rats injected with amphetamine (2mg/kg, i.p.) once daily for 3 consecutive days were then challenged with amphetamine (2mg/kg, i.p.) 5 days later. Expression of locomotor sensitization was blocked when methanandamide (5mg/kg, i.p.) was given once, just prior to amphetamine (2mg/kg, i.p.) challenge. In rats co-exposed to methanandamide (5mg/kg, i.p.) and amphetamine (2mg/kg, i.p.) on days 1-3 and then challenged with amphetamine (2mg/kg, i.p.) following 5 days of drug absence, the development of both locomotor and stereotypical sensitization was blocked. The ability of methanandamide to block amphetamine-sensitized behaviors suggests that this pharmacologically diverse lipid regulates signaling events impacted by repeated psychostimulant exposure.

Cine delayed-enhancement MR imaging of the heart: initial experience.

This study was performed by using an institutional review board-approved protocol, with waived informed consent and HIPAA compliance. The purpose of this study was to preliminarily evaluate a cine delayed-enhancement (DE) pulse sequence for depiction of wall motion and myocardial scar extent during a single acquisition. The technique is based on inversion-recovery single-shot balanced steady-state free precession magnetic resonance imaging. Cine DE images were acquired in 26 patients (18 men, eight women; age range, 25-84 years; mean age, 61 years+/-13 [standard deviation]). Image contrast was consistent throughout each series. Overall (ie, with both readers' scores averaged), the cine DE imaging-depicted wall motion was scored correctly in 71% of myocardial segments. Scar extent was scored correctly in 76% of segments; in no patient was scarring missed. Cine DE imaging is a promising technique for simultaneous visualization of wall motion and myocardial scar extent.

Pulsatile motion effects on 3D magnetic resonance angiography: implications for evaluating carotid artery stenoses.

In-plane carotid artery motion during a 3D MR angiography (MRA) scan can significantly degrade the resulting image resolution. This study characterizes the effect of cardiac pulsatility on 3D contrast-enhanced (CE) MRA with elliptical centric acquisitions using a point-spread function (PSF) analysis. Internal carotid artery (ICA) motion was collected from volunteers and patients using both MR and ultrasound (US) scans. After measuring the carotid artery motion displacement, a simulation was performed which calculated the blurring effects for three different protocols: nongated and two different cardiac gating schemes. The motion sensitivity of each protocol was evaluated for different spatial resolutions. The selection of optimal imaging parameters for a given scan time was investigated. The final results showed that cardiac-gated acquisitions only over a limited region of k-space high spatial frequencies are more time-efficient than cardiac gating for the entire k-space, as it allows for higher resolutions to be achieved and for capturing the arterial phase with low spatial frequencies. Selecting the optimal gating parameters depends directly on the motion characteristics of each individual. Our initial clinical experience is presented, and the need for a real-time tool that characterizes motion behavior for each individual as a prescan protocol is discussed.

Spinal dural arteriovenous fistula localization with a technique of first-pass gadolinium-enhanced MR angiography: initial experience.

Nine patients with initial magnetic resonance (MR) imaging and clinical findings suggestive of spinal dural arteriovenous fistula (AVF) underwent spinal MR angiography with an autotriggered elliptic centric ordered three-dimensional gadolinium-enhanced technique (hereafter, this MR angiographic technique) before conventional intraarterial angiography. In all nine patients, findings with this MR angiographic technique correctly and precisely localized the spinal dural AVF. Observer error resulted in one case in which the site of the fistula was not prospectively reported but was easily identified retrospectively on the spinal MR angiogram.