Acute and Repeated Administration of NLX-101, a Selective Serotonin-1A Receptor Biased Agonist, Reduces Audiogenic Seizures in Developing Fmr1 Knockout Mice.

Fragile XSyndrome (FXS) is a leading known genetic cause of Autism Spectrum Disorders (ASD) and intellectual disability. A consistent and debilitating phenotype of FXS is sensory hypersensitivity that manifests strongly in the auditory domain and may lead to delayed language and high anxiety. The mouse model of FXS, the Fmr1 KO mouse, also shows auditory hypersensitivity, an extreme form of which is seen as audiogenic seizures (AGS). The midbrain inferior colliculus (IC) is critically involved in generating audiogenic seizures and IC neurons are hyper-responsive to sounds in developing Fmr1 KO mice. Serotonin-1A receptor (5-HT1A) activation reduces IC activity. Therefore, we tested whether 5-HT1A activation is sufficient to reduce audiogenic seizures in Fmr1 KO mice. A selective and post-synaptic 5-HT1A receptor biased agonist, 3-Chloro-4-fluorophenyl-[4-fluoro-4-[[(5-methylpyrimidin-2-ylmethyl)amino]methyl]piperidin-1-yl] methanone (NLX-101, 0.6, 1.2, 1.8 or 2.4 mg/kg, i.p.) was administered to Fmr1 KO mice 15 min before seizure induction. Whereas the 0.6 mg/kg dose was ineffective in reducing seizures, the 1.2, 1.8 and 2.4 mg/kg doses of NLX-101 dramatically reduced seizures and increased mouse survival. Treatment with a combination of NLX-101 and 5-HT1A receptor antagonists prevented the protective effects of NLX-101, indicating that NLX-101 acts selectively through 5-HT1A receptors to reduce audiogenic seizures. NLX-101 (1.8 mg/kg) was still strongly effective in reducing seizures even after repeated administration over 5 days, suggesting an absence of tachyphylaxis to the effects of the compound. Together, these studies point to a promising treatment option targeting post-synaptic 5-HT1A receptors to reduce auditory hypersensitivity in FXS, and potentially across autism spectrum disorders.

Impact of aging on mitochondrial respiration in various organs.

Mitochondria are considered central regulator of the aging process; however, majority of studies dealing with the impact of age on mitochondrial oxygen consumption focused on skeletal muscle concluding (although not uniformly) a general declining trend with advancing age. In addition, gender related differences in mitochondrial respiration have not been satisfactorily described yet. The aim of the present study was to evaluate mitochondrial oxygen consumption in various organs of aging male and female Fischer 344 rats at the ages of 6, 12 and 24 months. Mitochondrial respiration of homogenized (skeletal muscle, left and right heart ventricle, hippocampus, cerebellum, kidney cortex), gently mechanically permeabilized (liver) tissue or intact cells (platelets) was determined using high-resolution respirometry (oxygraphs O2k, Oroboros, Austria). The pattern of age-related changes differed in each tissue: in the skeletal muscle and kidney cortex of both sexes and in female heart, parameters of mitochondrial respiration significantly declined with age. Resting respiration of intact platelets displayed an increasing trend and it did not correlate with skeletal muscle respiratory states. In the heart of male rats and brain tissues of both sexes, respiratory states remained relatively stable over analyzed age categories with few exceptions of lower mitochondrial oxygen consumption at the age of 24 months. In the liver, OXPHOS capacity was higher in females than in males with either no difference between the ages of 6 and 24 months or even significant increase at the age of 24 months in the male rats. In conclusion, the results of our study indicate that the concept of general pattern of age-dependent decline in mitochondrial oxygen consumption across different organs and tissues could be misleading. Also, the statement of higher mitochondrial respiration in females seems to be conflicting, since the gender-related differences may vary with the tissue studied, combination of substrates used and might be better detectable at younger ages than in old animals.

Age- and movement-related modulation of cortical oscillations in a mouse model of presbycusis.

Brain oscillations are associated with specific cognitive and sensory processes. How age-related hearing loss (presbycusis) alters cortical oscillations is unclear. Altered inhibitory neurotransmission and temporal processing deficits contribute to speech recognition impairments in presbycusis. Specifically, age-related reduction in parvalbumin positive interneurons and perineuronal nets in the auditory cortex predicts a reduction in gamma oscillations that may lead to a decline in temporal precision and attention. To test the hypothesis that resting and evoked gamma oscillations decline with presbycusis, EEGs were recorded from the auditory and frontal cortex of awake, freely moving C57BL/6 J mice at three ages (3, 14 and 24 months). Resting EEG data were analyzed according to movement state (move versus still). Evoked responses were recorded following presentation of noise bursts or amplitude modulated noise with time varying modulation frequencies. We report an age-related decrease in resting gamma power, a decline in gamma-range synchrony to time varying stimuli, and an increase in noise evoked and induced gamma power. A decline in temporal processing is seen in aged mice that exhibit robust auditory-evoked potentials, dissociating hearing loss from temporal processing deficits. We also report an increase in gamma power when mice moved compared to the still state. However, the movement-related modulation of gamma oscillations did not change with age. Together, these data identify a number of novel markers of presbycusis-related changes in auditory and frontal cortex. Because EEGs are commonly recorded in humans, the mouse data may serve as translation relevant preclinical biomarkers to facilitate the development of therapeutics to delay or reverse central auditory processing deficits in presbycusis.

TENS for surgical stabilisation of acetabular fracture in the skeletally immature: A novel technique.

BACKGROUND: Acetabular fractures in childhood are rare and the literature is scarce to describe a standard protocol in surgical management of these injuries. As the patient is still growing, it warrants a detailed assessment with a sound surgical plan if operative intervention is deemed necessary to prevent late complications. Throughout literature, most fixation rely on using pins, screws, plates or combination of the three which require large surgical exposure and risk of secondary physeal injury, hence we come up with a method of using the Titanium Elastic Nail System (TENS) to overcome this issue. We describe a novel technique in managing acetabular fractures in this group of patients using the TENS. METHOD: An 8 year old girl with a diagnosis of right anterior column posterior hemitransverse acetabular fracture was fixed with 3 TENS for supra-acetabular, anterior column and posterior column fragments. Surgery was performed in a minimally invasive manner. No drilling was performed during the surgery and implant insertion is done manually. RESULTS: Advantages of this procedure include minimally invasive surgery with smaller wounds, minimal intraoperative bleeding and theoretically reduces the risk of premature fusion of the triradiate cartilage. Patient is allowed early rehabilitation with this method. CONCLUSION: This novel method provides an alternative to traditional usage of wires, pins, plates and screws as is described in most literature. However, it requires the surgeon to appreciate that the safe corridors for the implant are much narrower than adults. We recommend this technique for fractures that are deemed suitable for intramedullary fixation and further research in the future will be needed.

Respiratory dysfunction in myotonic dystrophy type 1: A systematic review.

Myotonic dystrophy type 1 (DM1) is one of the most common muscular dystrophies in adults. This review summarises the current literature regarding the natural history of respiratory dysfunction in DM1, the role of central respiratory drive and peripheral respiratory muscle involvement and its significance in respiratory function, and investigates the relationship between genetics (CTG repeat length) and respiratory dysfunction. The review included all articles that reported spirometry on 10 or more myotonic dystrophy patients. The final review included 55 articles between 1964 and 2017. The major conclusions of this review were (1) confirmation of the current consensus that respiratory dysfunction, predominantly a restrictive ventilatory pattern, is common in myotonic dystrophy and is associated with alveolar hypoventilation, chronic hypercapnia, and sleep disturbance in the form of sleep apnoea and sleep related disordered breathing; (2) contrary to commonly held belief, there is no consensus in the literature regarding the relationship between CTG repeat length and severity of respiratory dysfunction and a relationship has not been established; (3) the natural history and time-course of respiratory functional decline is very poorly understood in the current literature; (4) there is a consensus that there is a significant involvement of central respiratory drive in this alveolar hypoventilation however the current literature does not identify the mechanism for this.

Sensory processing in autism spectrum disorders and Fragile X syndrome-From the clinic to animal models.

Brains are constantly flooded with sensory information that needs to be filtered at the pre-attentional level and integrated into endogenous activity in order to allow for detection of salient information and an appropriate behavioral response. People with Autism Spectrum Disorder (ASD) or Fragile X Syndrome (FXS) are often over- or under-reactive to stimulation, leading to a wide range of behavioral symptoms. This altered sensitivity may be caused by disrupted sensory processing, signal integration and/or gating, and is often being neglected. Here, we review translational experimental approaches that are used to investigate sensory processing in humans with ASD and FXS, and in relevant rodent models. This includes electroencephalographic measurement of event related potentials, neural oscillations and mismatch negativity, as well as habituation and pre-pulse inhibition of startle. We outline robust evidence of disrupted sensory processing in individuals with ASD and FXS, and in respective animal models, focusing on the auditory sensory domain. Animal models provide an excellent opportunity to examine common mechanisms of sensory pathophysiology in order to develop therapeutics.

Long-lasting effects of minocycline on behavior in young but not adult Fragile X mice.

Fragile X Syndrome (FXS) is the most common single-gene inherited form of intellectual disability with behaviors characteristic of autism. People with FXS display childhood seizures, hyperactivity, anxiety, developmental delay, attention deficits, and visual-spatial memory impairment, as well as a propensity for obsessive-compulsive disorder. Several of these aberrant behaviors and FXS-associated synaptic irregularities also occur in "fragile X mental retardation gene" knock-out (Fmr1 KO) mice. We previously reported that minocycline promotes the maturation of dendritic spines - postsynaptic sites for excitatory synapses - in the developing hippocampus of Fmr1 KO mice, which may underlie the beneficial effects of minocycline on anxiolytic behavior in young Fmr1 KO mice. In this study, we compared the effectiveness of minocycline treatment in young and adult Fmr1 KO mice, and determined the dependence of behavioral improvements on short-term versus long-term minocycline administration. We found that 4- and 8-week-long treatments significantly reduced locomotor activity in both young and adult Fmr1 KO mice. Some behavioral improvements persisted in young mice post-treatment, but in adults the beneficial effects were lost soon after minocycline treatment was stopped. We also show, for the first time, that minocycline treatment partially attenuates the number and severity of audiogenic seizures in Fmr1 KO mice. This report provides further evidence that minocycline treatment has immediate and long-lasting benefits on FXS-associated behaviors in the Fmr1 KO mouse model.

Parvalbumin immunoreactivity in the auditory cortex of a mouse model of presbycusis.

Age-related hearing loss (presbycusis) affects ∼35% of humans older than sixty-five years. Symptoms of presbycusis include impaired discrimination of sounds with fast temporal features, such as those present in speech. Such symptoms likely arise because of central auditory system plasticity, but the underlying components are incompletely characterized. The rapid spiking inhibitory interneurons that co-express the calcium binding protein Parvalbumin (PV) are involved in shaping neural responses to fast spectrotemporal modulations. Here, we examined cortical PV expression in the C57bl/6 (C57) mouse, a strain commonly studied as a presbycusis model. We examined if PV expression showed auditory cortical field- and layer-specific susceptibilities with age. The percentage of PV-expressing cells relative to Nissl-stained cells was counted in the anterior auditory field (AAF) and primary auditory cortex (A1) in three age groups: young (1-2 months), middle-aged (6-8 months) and old (14-20 months). There were significant declines in the percentage of cells expressing PV at a detectable level in layers I-IV of both A1 and AAF in the old mice compared to young mice. In layers V-VI, there was an increase in the percentage of PV-expressing cells in the AAF of the old group. There were no changes in percentage of PV-expressing cells in layers V-VI of A1. These data suggest cortical layer(s)- and field-specific susceptibility of PV+ cells with presbycusis. The results are consistent with the hypothesis that a decline in inhibitory neurotransmission, particularly in the superficial cortical layers, occurs with presbycusis.

Mechanisms underlying azimuth selectivity in the auditory cortex of the pallid bat.

This study focused on mechanisms underlying azimuth selectivity in the primary auditory cortex (A1) of pallid bats. The pallid bat listens to prey-generated noise (5-35 kHz) to localize and hunt terrestrial prey. The region of A1 tuned between 5 and 35 kHz consists of two clusters of neurons distinguished by interaural intensity difference (IID) selectivity: binaurally inhibited (EI) and peaked. The first aim of this study was to use sequential dichotic/free-field stimulation to test the hypothesis that IID is the primary cue underlying azimuth selectivity in neurons tuned in the prey-generated noise frequency band. IID selectivity and ear directionality at the neuron's characteristic frequency (CF) were used to predict azimuth selectivity functions. The predicted azimuth selectivity was compared with the actual azimuth selectivity from the same neurons. Prediction accuracy was similarly high for EI neurons and peaked neurons with low CF, whereas predictions were increasingly inaccurate with increasing CF among the peaked neurons. The second aim of this study was to compare azimuth selectivity obtained with noise and CF tones to determine the extent to which stimulus bandwidth influences azimuth selectivity in neurons with different binaural properties. The azimuth selectivity functions were similar for the two stimuli in the majority of EI neurons. A greater percentage of peaked neurons showed differences in their azimuth selectivity for noise and tones. This included neurons with multiple peaks when tested with tones and a single peak when tested with noise. Taken together, data from the two aims suggest that azimuth tuning of EI neurons is primarily dictated by IID sensitivity at CF. Peaked neurons, particularly those with high CF, may integrate IID sensitivity across frequency to generate azimuth selectivity for broadband sound. The data are consistent with those found in cat and ferret A1 in that binaurally facilitated neurons depend to a greater extent (compared to EI neurons) on spectral integration of binaural properties to generate azimuth selectivity for broadband stimuli.

Mechanisms underlying intensity-dependent changes in cortical selectivity for frequency-modulated sweeps.

Frequency-modulated (FM) sweeps are common components of species-specific vocalizations. The intensity of FM sweeps can cover a wide range in the natural environment, but whether intensity affects neural selectivity for FM sweeps is unclear. Bats, such as the pallid bat, which use FM sweeps for echolocation, are suited to address this issue, because the intensity of echoes will vary with target distance. In this study, FM sweep rate selectivity of pallid bat auditory cortex neurons was measured using downward sweeps at different intensities. Neurons became more selective for FM sweep rates present in the bat's echolocation calls as intensity increased. Increased selectivity resulted from stronger inhibition of responses to slower sweep rates. The timing and bandwidth of inhibition generated by frequencies on the high side of the excitatory tuning curve [sideband high-frequency inhibition (HFI)] shape rate selectivity in cortical neurons in the pallid bat. To determine whether intensity-dependent changes in FM rate selectivity were due to altered inhibition, the timing and bandwidth of HFI were quantified at multiple intensities using the two-tone inhibition paradigm. HFI arrived faster relative to excitation as sound intensity increased. The bandwidth of HFI also increased with intensity. The changes in HFI predicted intensity-dependent changes in FM rate selectivity. These data suggest that neural selectivity for a sweep parameter is not static but shifts with intensity due to changes in properties of sideband inhibition.

The Ghana community-based rehabilitation program for people with disabilities: what happened at the end of donor support?

In this case study the authors examined the functioning of the community-based rehabilitation (CBR) program for people with disabilities in 3 pilot districts after the conclusion of donor support in Ghana. Questionnaire and interview data from 42 people with disabilities, 8 local supervisors, and 3 social workers about program structures, support for people with disabilities, and challenges were analyzed using descriptive statistics and qualitative procedures, involving the use of Leximancer software. The authors found that some CBR structures remained in the communities. Diminished support for disabled peoples' organizations from communities and local government agencies were key challenges. The problem of volunteer local supervisors wanting to be paid ignited the evidence versus ideology debate around sustainability of CBR programs.

Visual experience is necessary for maintenance but not development of receptive fields in superior colliculus.

Sensory deprivation is thought to have an adverse effect on visual development and to prolong the critical period for plasticity. Once the animal reaches adulthood, however, synaptic connectivity is understood to be largely stable. We reported previously that N-methyl-D-aspartate (NMDA) receptor blockade in the superior colliculus of the Syrian hamster prevents refinement of receptive fields (RFs) in normal or compressed retinotopic projections, resulting in target neurons with enlarged RFs but normal stimulus tuning. Here we asked whether visually driven activity is necessary for refinement or maintenance of retinotopic maps or if spontaneous activity is sufficient. Animals were deprived of light either in adulthood only or from birth until the time of recording. We found that dark rearing from birth to 2 mo of age had no effect on the timing and extent of RF refinement as assessed with single unit extracellular recordings. Visual deprivation in adulthood also had no effect. Continuous dark rearing from birth into adulthood, however, resulted in a progressive loss of refinement, resulting in enlarged, asymmetric receptive fields and altered surround suppression in adulthood. Thus unlike in visual cortex, early visually driven activity is not necessary for refinement of receptive fields during development, but is required to maintain refined visual projections in adulthood. Because the map can refine normally in the dark, these results argue against a deprivation-induced delay in critical period closure, and suggest instead that early visual deprivation leaves target neurons more vulnerable to deprivation that continues after refinement.

Can two streams of auditory information be processed simultaneously? Evidence from the gleaning bat Antrozous pallidus.

A tenet of auditory scene analysis is that we can fully process only one stream of auditory information at a time. We tested this assumption in a gleaning bat, the pallid bat (Antrozous pallidus) because this bat uses echolocation for general orientation, and relies heavily on prey-generated sounds to detect and locate its prey. It may therefore encounter situations in which the echolocation and passive listening streams temporally overlap. Pallid bats were trained to a dual task in which they had to negotiate a wire array, using echolocation, and land on one of 15 speakers emitting a brief noise burst in order to obtain a food reward. They were forced to process both streams within a narrow 300 to 500 ms time window by having the noise burst triggered by the bats' initial echolocation pulses as it approached the wire array. Relative to single task controls, echolocation and passive sound localization performance was slightly, but significantly, degraded. The bats also increased echolocation interpulse intervals during the dual task, as though attempting to reduce temporal overlap between the signals. These results suggest that the bats, like humans, have difficulty in processing more than one stream of information at a time.

A systematic representation of interaural intensity differences in the auditory cortex of the pallid bat.

The current model of cortical processing of auditory spatial information is based on an orthogonal representation of frequency and binaural response properties, but how this arrangement leads to representation of space in the auditory cortex is unclear. This study describes the first evidence of a cortical substrate for the systematic representation of space in a region of primary auditory cortex of the pallid bat that subserves passive sound localization. The organizational feature of this region is a systematic shift in sensitivity to interaural intensity differences across the cortical surface, suggesting a topographic representation of horizontal space based on the distribution of activity within the neuron population.

Single cortical neurons serve both echolocation and passive sound localization.

The pallid bat uses passive listening at low frequencies to detect and locate terrestrial prey and reserves its high-frequency echolocation for general orientation. While hunting, this bat must attend to both streams of information. These streams are processed through two parallel, functionally specialized pathways that are segregated at the level of the inferior colliculus. This report describes functionally bimodal neurons in auditory cortex that receive converging input from these two pathways. Each brain stem pathway imposes its own suite of response properties on these cortical neurons. Consequently, the neurons are bimodally tuned to low and high frequencies, and respond selectively to both noise transients used in prey detection, and downward frequency modulation (FM) sweeps used in echolocation. A novel finding is that the monaural and binaural response properties of these neurons can change as a function of the sound presented. The majority of neurons appeared binaurally inhibited when presented with noise but monaural or binaurally facilitated when presented with the echolocation pulse. Consequently, their spatial sensitivity will change, depending on whether the bat is engaged in echolocation or passive listening. These results demonstrate that the response properties of single cortical neurons can change with behavioral context and suggest that they are capable of supporting more than one behavior.

TNF-mediated killing of human leukaemic cells: effects of endogenous antioxidant levels and TNF alpha expression in leukaemic cell lines.

Using the human erythroleukaemic cell line K562 cl.6 and its daunorubicin-resistant subline K/DAU600, and the human T-lymphoblastic leukaemic cell line CCRF-CEM and its vinblastine-resistant subline CEM/VLB100, we have shown that the drug-resistant cell lines were more sensitive to cytotoxicity induced by tumour necrosis factor-alpha (TNF alpha). Drug-resistant cell lines showed increased activities of copper/zinc superoxide dismutase (Cu/ZnSOD) and catalase compared with their parental drug-sensitive cell lines. However, the greater susceptibility of drug-resistant cells to TNF alpha cytotoxicity was, in part, related to their decreased activities of manganese superoxide dismutase (MnSOD). Persistence of this differential sensitivity when MnSOD is inhibited by sodium nitroprusside (SNP) suggests that the greater susceptibility of drug-resistant cells to TNF alpha was not entirely due to their decreased level of MnSOD activity. K562 cl.6 and K/DAU600, which were more resistant to TNF alpha, both expressed greater levels of endogenous plasma membrane-bound TNF alpha than the CCRF-CEM cell line. All cell lines examined were (more or less) equal in susceptibility to the cytolytic effect of exogenous O2-. generated by xanthine/xanthine oxidase. These results demonstrate that both MnSOD and endogenous TNF alpha play a role in protecting leukaemic cells against TNF alpha cytotoxicity, but there is an unknown mechanism that causes drug-resistant cells to be more susceptible to TNF alpha cytotoxicity.

Modulation of CD13 expression during retinoic acid-induced differentiation of HL60 cells.

CD13/aminopeptidase-N, an enzyme expressed by myeloid cells, may be important in the regulation and signalling pathways that control myeloid growth and differentiation. In this study we have used the myeloid leukaemic cell line HL60, and its ability to differentiate when induced by all-trans-retinoic acid (ATRA), to study the regulation of CD13 molecules, and its associated aminopeptidase-N enzyme activity during the myeloid differentiation pathway. In addition, the effect of the growth factor granulocyte-macrophage colony stimulating factor (GM-CSF) on CD13 expression, by undifferentiated and differentiated HL60 cells, has been investigated. Our results show that CD13 expression, and its enzyme activity, is downregulated during differentiation of HL60 induced by ATRA, but not when using GM-CSF.

Cyclosporine and cremaphor modulate von Willebrand factor release from cultured human endothelial cells.

Cyclosporine has been associated with microangiopathic hemolysis (MAHA) and other thrombotic complications of bone marrow and renal transplantation. MAHA is characterized by intravascular platelet aggregation, which, in some situations, is thought to be mediated by hyperactive high molecular weight von Willebrand factor (vWF). We have hypothesized that transplant-related MAHA may be caused by CsA-mediated release of von Willebrand factor from endothelial cells. This hypothesis was tested by studying vWF release from human umbilical vein endothelial cells primed with either CsA or cremophor EL. CsA and cremophor alone did not increase vWF release until toxic concentrations were reached (50-100 micrograms/ml). However, at therapeutic concentrations (0.1-5 micrograms/ml) vWF release by cells stimulated with thrombin, histamine, PMA, and the calcium ionophore A23187 was enhanced by both CsA and cremophor in a concentration-dependent manner. In single isolated endothelial cells, the thrombin-induced increase in cytosolic free calcium was enhanced by both CsA and cremophor. Preincubation for 24 hr with CsA but not cremophor suppressed vWF release after thrombin stimulation. These observations were mirrored by a concentration-dependent suppression of [3H]thymidine uptake by CsA. We conclude that CsA vehicle, cremophor, enhances stimulated vWF release in vitro, probably by processes dependent upon increased cytosolic free calcium. This suggests a possible mechanism for thrombotic transplant complications.

Induction of surface tumor necrosis factor (TNF) expression and possible facilitation of surface TNF release from human monocytic cells by granulocyte-macrophage colony-stimulating factor or gamma interferon in combination with 1,25-dihydroxyvitamin D3.

1,25-dihydroxyvitamin D3 (1,25(OH)2D3), gamma interferon (IFN-gamma) and granulocyte-macrophage colony-stimulating factor (GM-CSF) can regulate monocyte maturation and activation. Using the human monocytoid cell line U937, we have shown that these agents increase surface tumor necrosis factor (TNF) expression without directly affecting TNF release. GM-CSF and IFN-gamma combined with 1,25(OH)2D3 increased cellular TNF secretion to levels not seen with these agents alone. Ability to express and secrete TNF in part depended on degree of monocytic maturation. The combination of 1,25(OH)2D3 and GM-CSF, however, facilitated lipopolysaccharide (LPS)-mediated release of surface TNF from U937 cells, an effect that was temporally independent of maximal maturation. 1,25(OH)2D3 plus IFN-gamma was less effective than 1,25(OH)2D3 plus GM-CSF at facilitating TNF secretion. We postulate that 1,25(OH)2D3 and GM-CSF are required together to prime a specific mechanism, probably a protease, which cleaves TNF from the surface of monocytic cells. This protease, once primed, can be activated by a secondary stimulus such as LPS.