Applying computer-based language test to young children.

INTRODUCTION: This study aims at exploring the feasibility of applying a computer-based language test to young children aged 2-4 years. METHODS: Thirty-two Cantonese-speaking children, aged 2-4 years, were recruited from local kindergartens. All participants underwent assessment using both the computer-based and paper-pencil versions of the Macau Cantonese Language Screening Scale for Preschool Children, following a crossover study design. A short break of 15-30 minutes was provided between the two assessments. The data were analysed at three levels: the overall test, subcategory, and individual item levels. At the overall test and subcategory levels, data were analysed using the paired samples t-test and ICC. At the item level, the percentage of agreement and Cohen's kappa value were selected to assess the agreement of the two test formats. RESULTS: Excellent agreement was found for the overall test level, and good agreement was observed for four of the five subcategories. At the individual item level, 28 of the 35 items showed more than 80% agreement, and 16 items showed substantial to almost perfect agreement. CONCLUSION: These results suggest that the two test formats give similar total scores and subcategory scores for children aged 2-4. For children older than 2 years, 6 months, the agreement for matching items is as high as 83.68% (1318/1575). The computer-based test is thus highly recommended for this group of children. For children younger than 2 years, 6 months, a modified computer-based test is suggested to accommodate their needs.

The Impact of Maxillary Osteotomy on Fricatives in Cleft Lip and Palate: A Perceptual Speech and Acoustic Study.

INTRODUCTION: Abnormal facial growth is a recognized outcome in cleft lip and palate (CLP), resulting in a concave profile and a class III occlusal status. Maxillary osteotomy (MO) is undertaken to correct this facial deformity, and the surgery can impact speech articulation, although the evidence remains limited and ill-defined for the CLP population. AIMS: The aim of the study was to investigate the impact of MO on the production of the fricatives /f/ and /s/, using perceptual and acoustic analyses, and to explore the nature of speech changes. METHODS: Twenty participants with CLP were seen 0-3 months pre-operatively (T1) and 3 months (T2) and 12 months (T3) after MO. A normal group (N = 20) was similarly recruited. Perceptual speech data was collected according to a validated framework and ratings made on audio and audio-video recordings (VIDRat). Spectral moments were centre of gravity (CG), standard deviation (SD), skewness (SK) and kurtosis (KU). Reliability studies were carried out for all speech analyses. RESULTS: For the CLP group, VIDRat identified dentalization/interdentalization as the main type of pre-operative error for /s/ with a statistically significant improvement over time, χ2(2) = 6.889, p = 0.032. Effect sizes were medium between T1 and T3 (d = 0.631) and small between T2 and T3 (d = 0.194). For the acoustic data, effect sizes were similarly medium between T1 and T2 (e.g., SK, /f/ d = 0.579, /s/ d = 0.642) and small between T1 and T3 across all acoustic parameters. Independent t tests showed mainly statistically significant differences between both groups at all time points with large effect sizes (e.g., T2 CG, t = -4.571, p < 0.001, d =1.581), indicating that /s/ was not normalized post-operatively. For /f/, differences tended to be at T1 with large effect sizes (e.g., CG, t = -2.307, p = 0.028, d = 0.797), reflecting normalization. CONCLUSIONS AND IMPLICATIONS: This is the first speech acoustic study on /f/ for individuals with CLP undergoing MO. The surgery has a positive impact on /f/ and /s/, which appear to stabilize 3 months post-operatively. Speech changes are an automatic and a direct consequence of the physical changes brought about by MO, effecting articulatory re-organization. The results of the study have direct clinical implications for the clinical care pathway for patients with CLP undergoing MO.

Comparing Early Pragmatics in Typically Developing Children and Children with Neurodevelopmental Disorders.

This study examined the early pragmatic language skills in typically developing (TD) preschool-age children, children with language impairment (LI) and children with autism spectrum disorder (ASD). Two hundred and sixty-two TD children, 73 children with LI, and 16 children with ASD were compared on early pragmatics through direct assessment (DA). Post hoc analysis revealed that children in two clinical groups displayed significant pragmatic language deficits. Children in the ASD group who were older exhibited comparable degree of impairments as their LI peers, suggesting a relatively stagnant development of pragmatic language skills in children with ASD. Findings also supported the use of DA in identifying pragmatic language deficits, which have implications for the adoption of this assessment approach in clinical settings.

Are Vowels Normalized After Maxillary Osteotomy? An Acoustic Study in Cleft Lip and Palate.

BACKGROUND: Maxillary hypoplasia is a common skeletal condition in cleft lip and palate (CLP). Maxillary osteotomy is typically used to reposition the maxilla in CLP with maxillary hypoplasia. Previous studies have suggested that vowel articulations are adjusted postsurgically due to altered vocal tract configuration and articulatory reorganization. This acoustic study aims to investigate whether vowels are normalized postoperatively and to explore the nature of articulatory reorganization. METHODS AND PROCEDURES: A prospective study was conducted to examine the vowel production of a group of individuals with CLP (N = 17) undergoing maxillary osteotomy and a group of normal controls (N = 20), using speech acoustic data. The data were collected at 0 to 3 months presurgery (T1), 3-months (T2), and 12-months (T3) postsurgery. General linear model repeated measures and independent t-tests were undertaken on F1, F2, and vowel space area. RESULTS: General linear model repeated measures revealed no main effects of time for F1 (F [2, 22] = 1.094, P = 0.352), F2 (F [2, 22] = 1.269, P = 0.301), and vowel space area (F [2, 28] = 0.059, P = 0.943). Independent t-tests showed statistically significant differences (P < 0.05) for all acoustic parameters and all vowels between the CLP and the normal groups at all time points. CONCLUSIONS: Vowels were not normalized after maxillary osteotomy despite positive anatomical changes within the oral cavity. Individuals with CLP tended to adjust their vowel articulatory gestures to match presurgical patterns. The nature of articulatory reorganization appears to be prompt, sensory-driven, complete, and permanent.

Psychometric properties of the Chinese-version Stroke and Aphasia Quality of Life Scale 39-generic version (SAQOL-39g).

BACKGROUND AND PURPOSE: Aphasia is a common outcome of stroke affecting one-third of the post-stroke population in China. While the quality of life (QOL) may be affected, care is often inadequately guided due to lack of validated measure for Chinese population with stroke-induced aphasia. This study aimed to develop a Chinese-version of the Stroke and Aphasia Quality of Life-39 generic version (SAQOL-39g) and evaluate its feasibility, reliability, and validity in Chinese patients with stroke-induced aphasia. METHODS: The process of translation and adaptation suggested by WHO was used to develop the Chinese-version of SAQOL-39. We evaluated the feasibility, reliability, and validity of the scale in 84 aphasia patients and their proxies by assessing the internal consistency of the test items, test-retest consistency, and the structural validity of data. RESULTS: The self-report and the proxy-report form were completed within 21.4 and 13.3 min on average, respectively. Physical, communication, and psychological subdomains were extracted as three common factors. The Cronbach's alpha coefficients of overall domain and subdomains for both forms ranged from 0.879 to 0.950, indicating high internal consistency. The intraclass correlation coefficients ranged from 0.804 to 0.987 for overall domain and subdomains of the forms. No significant difference was found between two forms. CONCLUSIONS: The Chinese-version SAQOL-39g has excellent reliability, validity, and feasibility for measuring the QOL of Chinese post-stroke aphasia patients. The consistency between self-report and proxy-report forms was good, implying that the proxy-report form can be used to assess the QOL of post-stroke aphasia patients.

Ventral pallidum deep brain stimulation attenuates acute partial, generalized and tonic-clonic seizures in two rat models.

Approximately 30% of individuals with epilepsy are refractory to antiepileptic drugs and currently approved neuromodulatory approaches fall short of providing seizure freedom for many individuals with limited utility for generalized seizures. Here, we expand on previous findings and investigate whether ventral pallidum deep brain stimulation (VP-DBS) can be efficacious for various acute seizure phenotypes. For rats administered pilocarpine, we found that VP-DBS (50 Hz) decreased generalized stage 4/5 seizure median frequency from 9 to 6 and total duration from 1667 to 264 s even after generalized seizures emerged. The transition to brainstem seizures was prevented in almost all animals. VP-DBS immediately after rats exhibited their first partial forebrain stage 3 seizure did not affect the frequency of partial seizures but reduced median partial seizure duration from 271 to 54 s. Stimulation after partial seizures also reduced the occurrence and duration of secondarily generalized stage 4/5 seizures. VP-DBS prior to pilocarpine administration prevented the appearance of partial seizures in almost all animals. Lastly, VP-DBS delayed the onset of generalized tonic-clonic seizures (GTCSs) from 111 to 823 s in rats administered another chemoconvulsant, pentylenetetrazol (PTZ, 90 mg/kg). In this particular rat seizure model, stimulating electrodes placed more laterally in both VP hemispheres and more posterior in the left VP hemisphere provided greatest efficacy for GTCSs. In conclusion, our findings posit that VP-DBS can serve as an effective novel neuromodulatory approach for a variety of acute seizure phenotypes.

Deep Brain Stimulation of the Ventral Pallidum Attenuates Epileptiform Activity and Seizing Behavior in Pilocarpine-Treated Rats.

BACKGROUND: Brain stimulation is effective for people with intractable epilepsy. However, modulating neural targets that provide greater efficacy to more individuals is still needed. OBJECTIVE/HYPOTHESIS: We investigate whether bilateral deep brain stimulation of the ventral pallidum (VP-DBS) has potent seizure control in pilocarpine-treated rats. METHODS: VP-DBS (50 Hz) was applied prior to generalized forebrain seizures or after generalized brainstem seizures manifested. Behavioral seizures were assessed using a modified Racine scale. In vitro and in vivo electrophysiological techniques were employed to identify how VP-DBS affects proximal and distal neuronal activity. The open field test was used to see if acute and chronic VP-DBS affected gross motor function or arousal state. Parametric and non-parametric statistics with post-hoc analysis were performed. RESULTS: VP-DBS prior to pilocarpine prevented behavioral forebrain and brainstem seizures in most animals (n = 15). VP-DBS after brainstem seizures emerged prevented or reduced the appearance of subsequent behavioral brainstem seizures (n = 11). VP-DBS attenuated epileptiform activity in the hippocampus (n = 5), but not in the primary somatosensory cortex (S1) (n = 4) in vivo. Electrical stimulation in the VP increased VP GABAergic neuronal firing activity from 3.1 ± 1.4 Hz to 7.6 ± 1.7 Hz (n = 8) in vitro and reduced substantia nigra reticulata and superior colliculus neuronal spiking activity from 25.4 ± 3.3 Hz to 18.2 ± 1.4 Hz (n = 6) and 18.2 ± 1.4 Hz to 11.0 ± 1.1 Hz (n = 18), respectively, in vivo. CONCLUSION: VP-DBS can be a novel and potent therapeutic approach for individuals with intractable epilepsy.

Isovaline attenuates generalized epileptiform activity in hippocampal and primary sensory cortices and seizure behavior in pilocarpine treated rats.

Anti-seizure drugs are the most commonly employed treatment option for epilepsy and these generally provide effective management of seizures. However, 30% of patients with epilepsy are not adequately treated with anti-seizure medications and are considered intractable. Recently we reported that isovaline, a unique amino acid, could attenuate seizure like events (SLEs) in two in vitro hippocampal seizure models by selectively increasing the activity of interneurons, but not pyramidal neurons. Isovaline also attenuated hippocampal epileptiform activity and behavioral seizures in vivo in rats administered 4 aminopyridine (4AP). Here, we investigate whether isovaline is efficacious in attenuating secondarily generalized epileptiform activity and behavioral seizures in rats administered pilocarpine. We found that 150 mg/kg isovaline administered intravenously abolished pilocarpine-induced epileptiform activity in the primary sensory cortex and hippocampus and attenuated generalized forebrain behavioral seizures. We are the first to demonstrate that isovaline may be a plausible anti-seizure drug for secondarily generalized seizures and this could potentially lead to the development of a novel class of anti-seizure drugs focused around the unique mechanism(s) of isovaline.

Gap junction blockers attenuate beta oscillations and improve forelimb function in hemiparkinsonian rats.

Parkinson's disease (PD) is a neurodegenerative disease characterized by akinesia, bradykinesia, resting tremors and postural instability. Although various models have been developed to explain basal ganglia (BG) pathophysiology in PD, the recent reports that dominant beta (ß) oscillations (12-30Hz) in BG nuclei of PD patients and parkinsonian animals coincide with motor dysfunction has led to an emerging idea that these oscillations may be a characteristic of PD. Due to the recent realization of these oscillations, the cellular and network mechanism(s) that underlie this process remain ill-defined. Here, we postulate that gap junctions (GJs) can contribute to ß oscillations in the BG of hemiparkinsonian rats and inhibiting their activity will disrupt neuronal synchrony, diminish these oscillations and improve motor function. To test this, we injected the GJ blockers carbenoxolone (CBX) or octanol in the right globus pallidus externa (GPe) of anesthetized hemiparkinsonian rats and noted whether subsequent changes in ß oscillatory activity occurred using in vivo electrophysiology. We found that systemic treatment of 200mg/kg CBX attenuated normalized GPe ß oscillatory activity from 6.10±1.29 arbitrary units (A.U.) (pre-CBX) to 2.48±0.87 A.U. (post-CBX) with maximal attenuation occurring 90.0±20.5min after injection. The systemic treatment of octanol (350mg/kg) also decreased ß oscillatory activity in a similar manner to CBX treatment with ß oscillatory activity decreasing from 3.58±0.89 (pre-octanol) to 2.57±1.08 after octanol injection. Next, 1µl CBX (200mg/kg) was directly injected into the GPe of anesthetized hemiparkinsonian rats; 59.2±19.0min after injection, ß oscillations in this BG nucleus decreased from 3.62±1.17 A.U. to 1.67±0.62 A.U. Interestingly, we were able to elicit ß oscillations in the GPe of naive non-parkinsonian rats by increasing GJ activity with 1µl trimethylamine (TMA, 500nM). Finally, we systemically injected CBX (200mg/kg) into hemiparkinsonian rats which attenuated dominant ß oscillations in the right GPe and also improved left forepaw akinesia in the step test. Conversely, direct injection of TMA into the right GPe of naive rats induced contralateral left forelimb akinesia. Overall, our results suggest that GJs contribute to ß oscillations in the GPe of hemiparkinsonian rats.

Seizure-dependent mTOR activation in 5-HT neurons promotes autism-like behaviors in mice.

Epilepsy and autism spectrum disorder (ASD) are common comorbidities of one another. Despite the prevalent correlation between the two disorders, few studies have been able to elucidate a mechanistic link. We demonstrate that forebrain specific Tsc1 deletion in mice causes epilepsy and autism-like behaviors, concomitant with disruption of 5-HT neurotransmission. We find that epileptiform activity propagates to the raphe nuclei, resulting in seizure-dependent hyperactivation of mTOR in 5-HT neurons. To dissect whether mTOR hyperactivity in 5-HT neurons alone was sufficient to recapitulate an autism-like phenotype we utilized Tsc1flox/flox;Slc6a4-cre mice, in which mTOR is restrictively hyperactivated in 5-HT neurons. Tsc1flox/flox;Slc6a4-cre mice displayed alterations of the 5-HT system and autism-like behaviors, without causing epilepsy. Rapamycin treatment in these mice was sufficient to rescue the phenotype. We conclude that the spread of seizure activity to the brainstem is capable of promoting hyperactivation of mTOR in the raphe nuclei, which in turn promotes autism-like behaviors. Thus our study provides a novel mechanism describing how epilepsy can contribute to the development of autism-like behaviors, suggesting new therapeutic strategies for autism.

Cognitive Collaborations: Bidirectional Functional Connectivity Between the Cerebellum and the Hippocampus.

There is a growing recognition that the utility of the cerebellum is not limited to motor control. This review focuses on the particularly novel area of hippocampal-cerebellar interactions. Recent work has illustrated that the hippocampus and cerebellum are functionally connected in a bidirectional manner such that the cerebellum can influence hippocampal activity and vice versa. This functional connectivity has important implications for physiology, including spatial navigation and timing-dependent tasks, as well as pathophysiology, including seizures. Moving forward, an improved understanding of the critical biological underpinnings of these cognitive collaborations may improve interventions for neurological disorders such as epilepsy.

Isovaline attenuates epileptiform activity and seizure behavior in 4-aminopyridine treated rats.

Epilepsy is the most common neurological disorder in the world and although there are various treatment options available, 30% of patients remain intractable. Current antiepileptic drugs (AEDs) provide efficacy primarily by decreasing excitation or increasing inhibition in the seizing brain. Isovaline, a unique amino acid, was shown to attenuate seizure-like events (SLEs) in two in vitro hippocampal seizure models by selectively increasing the activity of interneurons, but not pyramidal neurons. Here, we demonstrate that 4-aminopyridine (4-AP) induced hippocampal epileptiform activity in vivo and seizing behavior, which were attenuated with intravenous (IV) isovaline treatment. We are the first to demonstrate that isovaline has potential as an AED and a conceptual framework for managing epilepsy could revolve around its novel mechanism of action.

Genomic and metabolic preparation of muscle in sockeye salmon Oncorhynchus nerka for spawning migration.

Prolonged endurance exercise and fasting are two major metabolic challenges facing Pacific salmon during spawning migrations that often occur over 1,000 km. Because both prolonged exercise and fasting stimulate the oxidation of lipids, particularly in heavily recruited tissues such as muscle, we sought to investigate the regulatory mechanisms that establish and maintain the capacity for substrate oxidation at four separate locations during the final 750 km of nonfeeding migration in sockeye salmon Oncorhynchus nerka. Transcript levels of multiple genes encoding for important regulators of lipid, carbohydrate, and protein oxidation as well as the activity of several important enzymes involved in lipid and carbohydrate oxidation were examined in red and white muscle. We found in both muscle types that the messenger RNA (mRNA) expression of carnitine palmitoyltransferase I isoforms, peroxisome proliferator-activated receptors α and ß, and adenosine monophosphate-activated protein kinase ß1 were all significantly higher at the onset compared to later stages of nonfeeding migration. However, the activities of ß-hydroxyacyl-CoA dehydrogenase and citrate synthase were higher only early in migration and only in red muscle. Later in the migration and as muscle lipid stores were greatly depleted, the mRNA levels of hexokinase I and aspartate aminotransferase increased in white muscle. Overall, at the onset of migration, high transcript and metabolic enzyme activity levels in skeletal muscle of sockeye salmon may help support the high rates of lipid oxidation needed for endurance swimming. Furthermore, we suggest that the muscle capacity to use carbohydrates and proteins may be adjusted throughout migration on an as-needed basis to fuel burst exercise through very difficult hydraulic passages in the river and perhaps during mating activities.

Elevated potassium provides an ionic mechanism for deep brain stimulation in the hemiparkinsonian rat.

The mechanism of high-frequency stimulation used in deep brain stimulation (DBS) for Parkinson's disease (PD) has not been completely elucidated. Previously, high-frequency stimulation of the rat entopeduncular nucleus, a basal ganglia output nucleus, elicited an increase in [K(+)](e) to 18 mm, in vitro. In this study, we assessed whether elevated K(+) can elicit DBS-like therapeutic effects in hemiparkinsonian rats by employing the limb-use asymmetry test and the self-adjusting stepping test. We then identified how these effects were meditated with in-vivo and in-vitro electrophysiology. Forelimb akinesia improved in hemiparkinsonian rats undergoing both tests after 20 mm KCl injection into the substantia nigra pars reticulata (SNr) or the subthalamic nucleus. In the SNr, neuronal spiking activity decreased from 38.2 ± 1.2 to 14.6 ± 1.6 Hz and attenuated SNr beta-frequency (12-30 Hz) oscillations after K(+) treatment. These oscillations are commonly associated with akinesia/bradykinesia in patients with PD and animal models of PD. Pressure ejection of 20 mm KCl onto SNr neurons in vitro caused a depolarisation block and sustained quiescence of SNr activity. In conclusion, our data showed that elevated K(+) injection into the hemiparkinsonian rat SNr improved forelimb akinesia, which coincided with a decrease in SNr neuronal spiking activity and desynchronised activity in SNr beta frequency, and subsequently an overall increase in ventral medial thalamic neuronal activity. Moreover, these findings also suggest that elevated K(+) may provide an ionic mechanism that can contribute to the therapeutic effects of DBS for the motor treatment of advanced PD.

Deep brain stimulation of the substantia nigra pars reticulata improves forelimb akinesia in the hemiparkinsonian rat.

Deep brain stimulation (DBS) employing high-frequency stimulation (HFS) is commonly used in the globus pallidus interna (GPi) and the subthalamic nucleus (STN) for treating motor symptoms of patients with Parkinson's disease (PD). Although DBS improves motor function in most PD patients, disease progression and stimulation-induced nonmotor complications limit DBS in these areas. In this study, we assessed whether stimulation of the substantia nigra pars reticulata (SNr) improved motor function. Hemiparkinsonian rats predominantly touched with their unimpaired forepaw >90% of the time in the stepping and limb-use asymmetry tests. After SNr-HFS (150 Hz), rats touched equally with both forepaws, similar to naive and sham-lesioned rats. In vivo, SNr-HFS decreased beta oscillations (12-30 Hz) in the SNr of freely moving hemiparkinsonian rats and decreased SNr neuronal spiking activity from 28 ± 1.9 Hz before stimulation to 0.8 ± 1.9 Hz during DBS in anesthetized animals; also, neuronal spiking activity increased from 7 ± 1.6 to 18 ± 1.6 Hz in the ventromedial portion of the thalamus (VM), the primary SNr efferent. In addition, HFS of the SNr in brain slices from normal and reserpine-treated rat pups resulted in a depolarization block of SNr neuronal activity. We demonstrate improvement of forelimb akinesia with SNr-HFS and suggest that this motor effect may have resulted from the attenuation of SNr neuronal activity, decreased SNr beta oscillations, and increased activity of VM thalamic neurons, suggesting that the SNr may be a plausible DBS target for treating motor symptoms of DBS.

Deconstructing the neural and ionic involvement of seizure-like events in the striatal network.

Seizures occur in the basal ganglia (BG) of epileptic patients and in animal models of epilepsy, but there is relatively little known about how these events are gated and/or propagated through this structure. Here, we present and characterize a model of in vitro seizure-like events (SLEs) in the striatum by applying chemostimulants to brain slices from young rat pups. We found that bath perfusion of artificial cerebral spinal fluid (aCSF) containing 0.25 mM MgCl(2), 5mM KCl and 100 µM 4-aminopyridine (LM/HK/4AP) elicited recurrent hyper-excitability in striatal medium spiny neurons (MSNs) in the form of paroxysmal depolarization shifts (PDSs) with an amplitude of 27.8 ± 2.1 mV and a duration of 29.4 ± 3.7s. PDSs coincided with SLEs in the striatal network with an amplitude of 106.5 ± 11.3 µV, duration of 23.6 ± 3.2 s, and a spiking frequency of 7.9 ± 1.3 Hz. Notably, chemostimulant-induced MSN PDSs were predominantly observed at earlier ages (P7-11), whereas occurrence of MSN PDSs declined to 50% by P12 and were no longer noted after P14; antagonism of the cannabinoid receptor (CB1) with 10 µM LY 320135 along with perfusion of LM/HK/4AP in older animals (P14-15) was unable to elicit MSN PDSs and SLEs. PDSs and SLEs were blocked with 60 µM 2-amino-5-phosphonopentanoate (APV), an N-methyl-d-aspartate receptor (NMDAR) blocker, or with traditional anticonvulsants such as 100 µM phenytoin or 50 µM carbamazepine. Conversely, blockade of 2-amino-3-(5-methyl-3-oxo-1,2-oxazol-4-yl) propanoic acid receptors (AMPARs) with 10 µM CNQX or T- and L-type Ca(2+) channels with 50 µM NiCl(2) or 50 µM nimodipine, respectively, did not significantly change MSN PDS and SLE amplitudes, durations and frequencies seen with LM/HK/4AP treatment alone. Striatal SLEs were driven by MSN hyper-excitability and synchrony since neither the presence of 1µM scopolamine, a muscarinic acetylcholine (ACh) receptor inhibitor, nor selective inhibition of fast-spiking interneurons (FSIs) with 50µM IEM1460 had any significant effect on MSN PDSs and SLEs. Next, we physically isolated the striatum from cortical and thalamic input and found that the striatum was intrinsically capable of manifesting NMDAR-dependent SLEs. Altogether, the present study is the first to deconstruct how SLEs can form in the striatum by examining how MSN activity coincides with SLEs. It also highlights a previously unrecognized potential for the striatum to manifest SLEs in vitro, without involving the cortex and thalamus. From these findings, further hypotheses can be developed for studying the BG's role in seizure generation and propagation, which may lead to novel pharmacological targets for the treatment of epilepsy.

Electrical stimulation of the insular region attenuates nicotine-taking and nicotine-seeking behaviors.

Pharmacological inactivation of the granular insular cortex is able to block nicotine-taking and -seeking behaviors in rats. In this study, we explored the potential of modulating activity in the insular region using electrical stimulation. Animals were trained to self-administer nicotine (0.03 mg/kg per infusion) under a fixed ratio-5 (FR-5) schedule of reinforcement followed by a progressive ratio (PR) schedule. Evaluation of the effect of stimulation in the insular region was performed on nicotine self-administration under FR-5 and PR schedules, as well on reinstatement of nicotine-seeking behavior induced by nicotine-associated cues or nicotine-priming injections. The effect of stimulation was also examined in brain slices containing insular neurons. Stimulation significantly attenuated nicotine-taking, under both schedules of reinforcement, as well as nicotine-seeking behavior induced by cues and priming. These effects appear to be specific to nicotine-associated behaviors, as stimulation did not have any effect on food-taking behavior. They appear to be anatomically specific, as stimulation surrounding the insular region had no effect on behavior. Stimulation of brain slices containing the insular region was found to inactivate insular neurons. Our results suggest that deep brain stimulation to modulate insular activity should be further explored.

Identification and validation of hits from high throughput screens for CFTR modulators.

These are exciting times with the appearance of small molecule compounds in clinical trials which target the basic defects caused by mutation in the CFTR gene. This progress was enabled by years of basic research probing the molecular and cellular consequences caused by mutation and the development of methods by which to study the primary anion transport defect in a high-throughput manner by robotics. Future progress with the development of new, more effective corrector compounds is needed. Such discovery will require further progress in defining the molecular targets for effective intervention using a multidisciplinary approach, merging computational, molecular, proteomic and cell biological methods. There is also an urgent need to develop means to link the right therapeutic compound to the right patients given the heterogeneity of the CF patient population. We envision a time when mid to high-throughput methods will be married with stem cell biology to enable testing a compendium of compounds on cells derived from each individual patient. Given the rate of progress in this field- this scenario may exist in the not too distant future.

Synthesis and properties of molecular probes for the rescue site on mutant cystic fibrosis transmembrane conductance regulator.

Cystic fibrosis is a genetic disease caused by mutations in the gene for the cystic fibrosis transmembrane conductance regulator (CFTR) protein. In vitro experiments have demonstrated that 4-methyl-2-(5-phenyl-1H-pyrazol-3-yl)phenol (VRT-532, 1) is able to partially restore the function of mutant CFTR proteins. To help elucidate the nature of the interactions between 1 and mutant CFTR, molecular probes based on the structure of 1 have been prepared. These include a photoreactive aryl azide derivative 11 and a fluorescent dansyl sulfonamide 15. Additionally, a method for hydrogen isotope exchange on 1 has been developed, which could be used for the incorporation of radioactive tritium. Using iodide efflux assays, the probe molecules have been demonstrated to modulate the activity of mutant CFTR in the same manner as 1. These probe molecules enable a number of biochemical experiments aimed at understanding how 1 rescues the function of mutant CFTR. This understanding can in turn aid in the design and development of more efficacious compounds which may serve as therapeutic agents in the treatment of cystic fibrosis.