Development of a nanomaterial bio-screening platform for neurological applications

Nanoparticle platforms are being intensively investigated for neurological applications. Current biological models used to identify clinically relevant materials have major limitations, e.g. technical/ethical issues with live animal experimentation, failure to replicate neural cell diversity, limited control over cellular stoichiometries and poor reproducibility. High-throughput neuro-mimetic screening systems are required to address these challenges. We describe an advanced multicellular neural model comprising the major non-neuronal/glial cells of the central nervous system (CNS), shown to account for ~99.5% of CNS nanoparticle uptake. This model offers critical advantages for neuro-nanomaterials testing while reducing animal use: one primary source and culture medium for all cell types, standardized biomolecular corona formation and defined/reproducible cellular stoichiometry. Using dynamic time-lapse imaging, we demonstrate in real-time that microglia (neural immune cells) dramatically limit particle uptake in other neural subtypes (paralleling post-mortem observations after nanoparticle injection in vivo), highlighting the utility of the system in predicting neural handling of biomaterials

Electrophysiological Assessment of Primary Cortical Neurons Genetically Engineered using Iron Oxide Nanoparticles

The development of safe technologies to genetically modify neurons is of great interest in regenerative neurology, for both translational and basic science applications. Such approaches have conventionally been heavily reliant on viral transduction methods, which have safety and production limitations. Magnetofection (magnet-assisted gene transfer using iron oxide nanoparticles as vectors) has emerged as a highly promising non-viral alternative for safe and reproducible genetic modification of neurons. Despite the high potential of this technology, there is an important gap in our knowledge of the safety of this approach, namely, whether it alters neuronal function in adverse ways, such as by altering neuronal excitability and signaling. We have investigated the effects of magnetofection in primary cortical neurons by examining neuronal excitability using the whole cell patch clamp technique. We found no evidence that magnetofection alters the voltage-dependent sodium and potassium ionic currents that underpin excitability. Our study provides important new data supporting magnetofection as a safe technology for bioengineering of neuronal cell populations

Differences in magnetic particle uptake by CNS neuroglial subclasses: implications for neural tissue engineering

AIM: To analyze magnetic particle uptake and intracellular processing by the four main non-neuronal subclasses of the CNS: oligodendrocyte precursor cells; oligodendrocytes; astrocytes; and microglia. MATERIALS & METHODS: Magnetic particle uptake and processing were studied in rat oligodendrocyte precursor cells and oligodendrocytes using fluorescence and transmission electron microscopy, and the results collated with previous data from rat microglia and astrocyte studies. All cells were derived from primary mixed glial cultures. RESULTS: Significant intercellular differences were observed between glial subtypes: microglia demonstrate the most rapid/extensive particle uptake, followed by astrocytes, with oligodendrocyte precursor cells and oligodendrocytes showing significantly lower uptake. Ultrastructural analyses suggest that magnetic particles are extensively degraded in microglia, but relatively stable in other cells. CONCLUSION: Intercellular differences in particle uptake and handling exist between the major neuroglial subtypes. This has important implications for the utility of the magnetic particle platform for neurobiological applications including genetic modification, transplant cell labeling and biomolecule delivery to mixed CNS cell populations

2D non-LTE modelling of a filament observed in the H_alpha line with the DST/IBIS spectropolarimeter

We study a fragment of a large quiescent filament observed on May 29, 2017 by the Interferometric BIdimensional Spectropolarimeter (IBIS) mounted at the Dunn Solar Telescope. We focus on its quiescent stage prior to its eruption. We analyse the spectral observations obtained in the H$\alpha$ line to derive the thermodynamic properties of the plasma of the observed fragment of the filament. We used a 2D filament model employing radiative transfer computations under conditions that depart from the local thermodynamic equilibrium. We employed a forward modelling technique in which we used the 2D model to producesynthetic H_alpha line profiles that we compared with the observations. We then found the set of model input parameters, which produces synthetic spectra with the best agreement with observations. Our analysis shows that one part of the observed fragment of the filament is cooler, denser, and more dynamic than its other part that is hotter, less dense, and more quiescent. The derived temperatures in the first part range from 6,000 K to 10,000$ K and in the latter part from 11,000 K to 14,000 K. The gas pressure is 0.2-0.4 dyn/cm}^{2} in the first part and around 0.15 dyn/cm}^{2} in the latter part. The more dynamic nature of the first part is characterised by the line-of-sight velocities with absolute values of 6-7 km/s and microturbulent velocities of 8-9 km/s. On the other hand, the latter part exhibits line-of-sight velocities with absolute values 0-2.5 km/s and microturbulent velocities of 4-6 km/s

Identifying the cellular targets of drug action in the central nervous system following corticosteroid therapy

Corticosteroid (CS) therapy is used widely in the treatment of a range of pathologies, but can delay production of myelin, the insulating sheath around central nervous system nerve fibers. The cellular targets of CS action are not fully understood, that is, "direct" action on cells involved in myelin genesis [oligodendrocytes and their progenitors the oligodendrocyte precursor cells (OPCs)] versus "indirect" action on other neural cells. We evaluated the effects of the widely used CS dexamethasone (DEX) on purified OPCs and oligodendrocytes, employing complementary histological and transcriptional analyses. Histological assessments showed no DEX effects on OPC proliferation or oligodendrocyte genesis/maturation (key processes underpinning myelin genesis). Immunostaining and RT-PCR analyses show that both cell types express glucocorticoid receptor (GR; the target for DEX action), ruling out receptor expression as a causal factor in the lack of DEX-responsiveness. GRs function as ligand-activated transcription factors, so we simultaneously analyzed DEX-induced transcriptional responses using microarray analyses; these substantiated the histological findings, with limited gene expression changes in DEX-treated OPCs and oligodendrocytes. With identical treatment, microglial cells showed profound and global changes post-DEX addition; an unexpected finding was the identification of the transcription factor Olig1, a master regulator of myelination, as a DEX responsive gene in microglia. Our data indicate that CS-induced myelination delays are unlikely to be due to direct drug action on OPCs or oligodendrocytes, and may occur secondary to alterations in other neural cells, such as the immune component. To the best of our knowledge, this is the first comparative molecular and cellular analysis of CS effects in glial cells, to investigate the targets of this major class of anti-inflammatory drugs as a basis for myelination deficits

Exertional rhabdomyolysis in a 21-year-old, healthy female after performing three sets of the biceps curl exercise to failure with 30% 1RM: A case report

Background: The optimal resistance training program to elicit muscle hypertrophy has been consistently debated and researched. Although 3 sets of 10 repetitions at 70-80% of the 1-repetition maximum (1RM) is widely recommended, recent studies have shown that low-load (~30% 1RM), high-repetition (3 sets of 30-40 repetitions) resistance training can elicit similar muscular hypertrophy. Therefore, this type of resistance training has gained popularity, perhaps because less weight is lifted for a longer duration. In the process of testing this hypothesis in a research study in our laboratory, a subject diagnosed with exertional rhabdomyolysis after completing a single resistance training session that involved 3 sets to failure at 30% 1RM. Exertional rhabdomyolysis is a condition characterized by the excessive breakdown of striated skeletal muscle that releases proteins from the muscle cell, particularly myoglobin, into the blood that can be toxic to the kidneys and is a significant health concern. Case Report: Reviewed were the events leading up to and throughout the diagnosis of exertional rhabdomyolysis in a healthy, recreationally-trained, 21-year-old female that was enrolled in a study that compared the acute effects of the traditional high-load, low-repetition versus low-load, high-repetition resistance training. The subject completed a total of 143 repetitions of the bilateral dumbbell bicep curl exercise. Three days post-exercise she reported excessive muscle soreness and swelling and sought medical attention. She was briefly hospitalized and then discharged with instructions to take acetaminophen for soreness, drink plenty of water, rest, and monitor her creatine kinase (CK) concentrations. Changes in the subject’s CK concentrations, ultrasound-determined muscle thickness and echo intensity were monitored over a 14-day period are reported. Discussion: This case illustrates the potential risk of developing exertional rhabdomyolysis after a low-load, high-repetition resistance training session in healthy, young, recreationally-trained women. The fact that exertional rhabdomyolysis is a possible outcome is enough to warrant caution when prescribing this type of resistance exercise

Symptoms predicting psychosocial impairment in bulimia nervosa

Purpose The current study aimed to determine which particular eating disorder (ED) symptoms and related features, such as BMI and psychological distress, uniquely predict impairment in bulimia nervosa (BN). Methods Two hundred and twenty-two adults with BN completed questionnaires assessing ED symptoms, general psychological distress, and psychosocial impairment. Regression analyses were used to determine predictors which account for variance in impairment. Results Four variables emerged as significant predictors of psychosocial impairment: concerns with eating; concerns with weight and shape; dietary restraint; and general psychological distress. Conclusions Findings support previous work highlighting the importance of weight and shape concerns in determining ED-related impairment. Other ED symptoms, notably dietary restraint and concerns with eating, were also significant predictors as was psychological distress. Results suggest that cognitive aspects of EDs, in addition to psychological distress, may be more important determinants of impairment than behavioural symptoms, such as binge eating or purging

Contribution of the cyclic nucleotide gated channel subunit, CNG-3, to olfactory plasticity in Caenorhabditis elegans.

In Caenorhabditis elegans, the AWC neurons are thought to deploy a cGMP signaling cascade in the detection of and response to AWC sensed odors. Prolonged exposure to an AWC sensed odor in the absence of food leads to reversible decreases in the animal's attraction to that odor. This adaptation exhibits two stages referred to as short-term and long-term adaptation. Previously, the protein kinase G (PKG), EGL-4/PKG-1, was shown necessary for both stages of adaptation and phosphorylation of its target, the beta-type cyclic nucleotide gated (CNG) channel subunit, TAX-2, was implicated in the short term stage. Here we uncover a novel role for the CNG channel subunit, CNG-3, in short term adaptation. We demonstrate that CNG-3 is required in the AWC for adaptation to short (thirty minute) exposures of odor, and contains a candidate PKG phosphorylation site required to tune odor sensitivity. We also provide in vivo data suggesting that CNG-3 forms a complex with both TAX-2 and TAX-4 CNG channel subunits in AWC. Finally, we examine the physiology of different CNG channel subunit combinations

The effects of anatabine on non-invasive indicators of muscle damage: a randomized, double-blind, placebo-controlled, crossover study

Background: Anatabine (ANA), a minor tobacco alkaloid found in the Solanaceae family of plants, may exhibit anti-inflammatory activity, which may be useful to aid in recovery from exercise-induced muscle damage. The purpose of this study, therefore, was to examine the effects of ANA supplementation on the recovery of isometric strength and selected non-invasive indicators of muscle damage. Methods: A double-blinded, placebo-controlled, crossover design was used to study eighteen men (mean ± SD age = 22.2 ± 3.1 yrs; body mass = 80.3 ± 15.7 kg) who participated in two randomly-ordered conditions separated by a washout period. The ANA condition consisted of consuming 6–12 mg anatabine per day for 10 days, while testing took place during days 7–10. The placebo (PLA) condition was identical except that the PLA supplement contained no ANA. Maximal voluntary isometric peak torque (PT) of the forearm flexors, arm circumference, hanging joint angle, and subjective pain ratings were measured before (PRE), immediately after (POST), and 24, 48, and 72 h after six sets of 10 maximal, eccentric isokinetic forearm flexion muscle actions. Resting heart rate and blood pressure were measured at PRE and 72 h in each condition. Results: For PT, hanging joint angle, arm circumference, and subjective pain ratings, there were no condition x time (p \u3e 0.05) interactions, there were no main effects for condition (p \u3e 0.05), but there were main effects for time (p \u3c 0.001). There were no condition x time (p \u3e 0.05) interactions and no main effects for condition (p \u3e 0.05) or time (p \u3e 0.05) for blood pressure or resting heart rate. Conclusions: ANA supplementation had no effect on the recovery of muscle strength, hanging joint angle, arm swelling, or subjective pain ratings after a bout of maximal eccentric exercise in the forearm flexors. Therefore, ANA may not be beneficial for those seeking to improve recovery from heavy eccentric exercise. Future studies should examine the effects of ANA on the pro-inflammatory cytokine responses to exercise-induced muscle damage and the chronic low-grade inflammation observed in obese and elderly individuals

Exceptional sperm cooperation in the wood mouse

Spermatozoa from a single male will compete for fertilization of ova with spermatozoa from another male when present in the female reproductive tract at the same time. Close genetic relatedness predisposes individuals towards altruism, and as haploid germ cells of an ejaculate will have genotypic similarity of 50%, it is predicted that spermatozoa may display cooperation and altruism to gain an advantage when inter-male sperm competition is intense. We report here the probable altruistic behaviour of spermatozoa in an eutherian mammal. Spermatozoa of the common wood mouse, Apodemus sylvaticus, displayed a unique morphological transformation resulting in cooperation in distinctive aggregations or 'trains' of hundreds or thousands of cells, which significantly increased sperm progressive motility. Eventual dispersal of sperm trains was associated with most of the spermatozoa undergoing a premature acrosome reaction. Cells undergoing an acrosome reaction in aggregations remote from the egg are altruistic in that they help sperm transport to the egg but compromise their own fertilizing ability