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Background Late-onset Pompe disease (LOPD) is a recessive disease caused by α-glucosidase (GAA) deficiency, leading to progressive muscle weakness and/or respiratory failure in children and adults. Respiratory derangement can be the first indication of LOPD, but the diagnosis may be difficult for pneumologists. We hypothesize that assessing the GAA activity in suspected patients by a dried blood spot (DBS) may help the diagnosis of LOPD in the pneumological setting. Population and methods We performed a multicenter DBS survey of patients with suspected LOPD according to a predefined clinical algorithm. From February 2015 to December 2017, 140 patients (57 ± 16 yrs., 80 males) were recruited in 19 Italian pneumological units. The DBS test was performed by a drop of blood collected on absorbent paper. Patients with GAA activity

The aim of present research is to study the temporal structure of the behavior in two strains of rats with different basal level of emotionality. To this purpose, the temporal profile of the behavior in Wistar rat and in the spontaneously more anxious DA/Han strain was analyzed in the Elevated Plus Maze. Both quantitative and multivariate t-pattern analyses were carried out. In comparison with Wistar, DA/Han subjects showed a significant reduction of the permanence in open arm and a significant increase of the time spent in the central platform of the maze. Mean frequencies of each behavioral element showed significant modifications both in open and in closed arm. Multivariate t-pattern analyses demonstrated a very different temporal profile of behavior in the two strains: Wistar rats presented 197 t-patterns of different composition, whereas DA/Han rats only 26; as to the mean number of t-patterns, Wistar presented a value of 698.90, whereas DA/Han only 92.80. Similar clear-cut differences were detected for the mean number of t-patterns in the two arms and for the time course of such t-patterns. Present study has evidenced that the temporal organization of the bahavior in Elevated Plus Maze is differently structured in two strains of rats with different basal level of emotionality.

The objective of this study was to describe a large Italian cohort of patients with late-onset glycogen storage disease type 2 (GSDII) at various stages of disease progression and to evaluate the clinical effectiveness of alglucosidase alpha enzyme replacement therapy (ERT). Previous studies showed in late-onset patients ERT efficacy against placebo and variable response in uncontrolled studies. Seventy-four juvenile or adult GSDII patients were treated with ERT in a multicenter open label, non-randomized study, from 12 months up to 54 months. Recombinant human alpha glucosidase (rh-GAA) was injected by intravenous route at 20 mg/kg every second week. Patients were divided into three groups according to ERT duration: Group A received treatment for 12–23 months (n = 16), Group B for 24–35 months (n = 14), and Group C for more than 36 months (n = 44). Clinical assessment included a 6-min walk test (6MWT), forced vital capacity (FVC), the Walton and Gardner-Medwin score, the number of hours of ventilation, body mass index, echocardiography and blood creatine kinase (CK). Included in our cohort were 33 males and 41 females (M:F = 0.8:1), with a mean age at first symptoms of 28.3 years (range 2–55 years) and a mean age of 43 years at study entry (range 7–72 years). Seven wheelchair bound patients, as well as 27 patients requiring ventilation support, were included. After treatment we could observe an increase in distance walked on the 6MWT in the large majority of patients (48/58; 83%), with an overall mean increase of 63 m (from 320 ± 161 to 383 ± 178 m). After treatment in the majority of patients FVC was improved or unchanged (45/69; 65%). In ventilated patients we observed an improvement in average number of hours off the ventilator (from 15.6 to 12.1 h). Six patients stopped mechanical ventilation and two others started it. The effect of therapy was not related to ERT duration. Nine of 64 patients (13%) that underwent to echocardiography showed a variable degree of cardiac hypertrophy (left ventriculum or septum), and a positive effect was observed after 36 months of ERT in one adult case. Discontinuation of treatment occurred in four patients: one drop-off case, one patient died for a sepsis after 34 months of treatment and two patients stopped ERT for worsening of general clinical condition. Mild adverse effects were observed in four cases (5%). This study represents the largest cohort of late-onset GSDII patients treated with ERT, and confirm a positive effect of treatment. These results, obtained in a large case series on therapy, indicate a favourable effect of ERT therapy, even in more advanced stage of the disease.

Introduction: The clinical course of late-onset Pompe disease is heterogeneous, and new clinical outcome measures are needed to evaluate enzyme replacement therapy (ERT). Methods: We correlated the 6-Minute Walk Test (6MWT), Walton and Gardner-Medwin (WGM) score, and GSGC (Gait, Stairs, Gower, Chair) scores in 40 patients. Results: At baseline, the GSGC score correlated with both WGM (P < 0.001, n = 33) and 6MWT (P < 0.001, n = 26). After 1 year of ERT, we observed a significant change in gait, stairs and chair performance on the GSGC scale. The 6MWT significantly increased from 319 to 371 meters in 32 patients, and the WGM score was reduced. Conclusions: GSGC is a group of functional tests that requires only a few minutes to perform, therefore, this score might be a good indicator to be used in future studies. Muscle Nerve 45: 831–834, 2012

The influence of the A10 region of the ventral tegmental area (VTA) on the quiet biting attack evoked by stimulation of the lateral hypothalamic nucleus (LH) was studied. The latency of the biting was considered as reference value and measured with stop-watches; it remained constant when hypothalamic stimulation was performed with the same parameters. Simultaneous activation of the A10 neuron group induced a facilitation of the aggression in the form of a decrease in the biting latency or a display of the attack pattern when LH was stimulated with parameters below the threshold for biting appearance. The facilitatory effect of the A10 neurons of the VTA is discussed.

The effects of substantia nigra (SN) (pars compacta) stimulation on the rage reaction elicited by ventromedial hypothalamic nucleus (VMH) were investigated in the cat. The studied parameters of the rage reaction were: the current and the frequency threshold for the appearance of the hissing and the hissing latency. A facilitatory effect induced by the SN on the hypothalamic rage reaction was observed in the form of a decrease in the hypothalamic stimulus threshold for the hissing appearance and a decrease in the hissing latency. Moveover, when the VMH was stimulated with parameters below the threshold for the hissing display, simultaneous nigral activation determined its appearance. The excitatory influence exerted by the SN on the affective component of the aggressive behavior is discussed.

Publisher Summary This chapter explains the modifications of the spontaneous bioelectric activity and of the after discharge (AD) evoked in the amygdala after pallidal injection of kainic acid (KA) in the cat. An experiment was conducted in which injection of KA, a neurotoxic drug analog of glutamate, was made into the entopeduncolar nucleus (EN), the equivalent of the medial globus pallidus of primates, to observe the modifications produced on the spontaneous amygdaloid bioelectric activity and on the evoked AD. An AD was evoked in the amygdala by stimulation of the ipsilateral pyriform cortex. The duration of the amygdaloid AD was related to the stimulus parameters. In the same animal, the AD duration did not greatly change when stimulation parameters remained constant. Stereotaxically microinjection of KA was effected in the EN ipsilateral to the amygdaloid deriving electrodes. Few minutes after the injection of the drug, a decrease of the AD duration was observed likely linked to the excitatory effect of KA on the EN neurons that increased its inhibitory effect on the amygdaloid complex. The results of the experiment contribute to underline excitatory effect and subsequently damaging of KA on EN neurons allowing one to confirm a control role of the globus pallidus on the spontaneous and evoked paroxystic amygdaloid activity.

Tobacco smoking is a serious health problem worldwide and a leading cause of mortality. Nicotine, the addictive component of tobacco, affects a range of emotional responses, including anxiety-related behaviors. Although perceived by smokers to be anxiolytic, evidence suggests that smoking increases anxiety and that mood fluctuates with nicotine intake. Thus, nicotine addiction may depend on easing the psychobiological distress caused by its abuse. The lateral habenula (LHb) has been implicated as a neural substrate for acute nicotine-induced anxiety, but its role in anxiety-like behaviors associated with chronic nicotine exposure has not been explored. Here, we assessed the effect of chronic nicotine exposure and its subsequent overnight withdrawal on anxiety-like behavior using both quantitative and multivariate T-pattern analysis in rats tested using the hole-board apparatus. Additionally, we explored the role of the LHb by comparing the behavioral effects of short-term nicotine withdrawal in chronically treated LHb-lesioned rats. Quantitative analysis revealed increased anxiety-like behavior in chronically treated overnight nicotine-deprived rats, as manifested in reduced general and focused exploratory behaviors, which was eased in animals that received nicotine. Quantitative analysis failed to reveal a role of the LHb in overnight nicotine deprivation-induced anxiety. Conversely, T-pattern analysis of behavioral outcomes revealed that chronic nicotine-treated rats still show anxiety-like behavior following nicotine challenge. Moreover, it demonstrated that the LHb lesion induced a stronger anxiolytic-like response to the acute challenge of nicotine in chronically nicotine-exposed animals, implicating the LHb in the anxiogenic effect of chronic nicotine exposure. These data further highlight the LHb as a promising target for smoking cessation therapies and support the importance of T-pattern analysis for behavioral analysis.

Stiff person syndrome (SPS) is a rare neurologic disorder characterized by progressively worsening rigidity and spasms of the axial and limb muscles. Dyspnea has been recently recognized as a common symptom in SPS,1 and life-threatening respiratory crises have been occasionally reported and suspected to be responsible for sudden death in these patients.2,3 The pathophysiologic mechanisms of these respiratory manifestations remain unclear. Some authors have hypothesized that rigidity and/or spasm of the muscles of the trunk could prevent normal rib cage movements and excursion of the diaphragm.1

Little is known about the structural characteristics of the behavior of rats with enhanced anxiety level. To fill this gap, a study was undertaken where effects of an anxiogenic drug were examined on behavioral structure of rats tested in hole board.This study investigates effects of increased anxiety level on the structure of the behavior of rats tested in hole board METHODS: Different doses of FG7142 (1, 4, 8 mg/kg IP), a potent anxiety-inducing drug, were administered to three groups of male Wistar rats. A further group was administered saline. Experiments were recorded through a digital camera. Quantitative and multivariate approaches were applied.Percent distributions and durations showed increases of immobile sniffing, rearing, head dip, and edge sniff and a significant reduction of grooming activities and of walking. In addition, a decrease of head dip/edge sniff ratio was detected. Transition matrices evidenced that FG7142 provoked evident modifications of behavioral structure mainly of general exploration of environment and focused exploration of the hole. Finally, adjusted residuals showed a reduced effectiveness of FG7142 on transitions from head dip to edge sniff; on the contrary, transitions from edge sniff to head dip underwent evident dose-dependent changes.Present study provides a useful tool to analyze behavioral responses to different anxiety conditions. Accordingly, it is demonstrated that a condition of increased anxiety deeply modifies the structure of male Wistar rat's behavior in hole board. In addition, our results suggest that evaluation of head dip/edge sniff ratio can be considered a reliable index to appraise effects of pharmacological manipulation of anxiety and related behavioral elements.

Introduction Respiratory failure is the leading cause of death in amyotrophic lateral sclerosis (ALS) [1]. The involvement of respiratory function is a negative predictor of survival, which can be in part overcome by a prompt initiation of non-invasive mechanical ventilation (NIV). Seated forced vital capacity (FVC%) is a widely used measure to monitor respiratory function, and it is often taken as a surrogate outcome measure in clinical trials. However, it may be within a normal range in ALS patients with respiratory dysfunction [2]. Here we report on a case of an ALS patient with a rapidly evolving respiratory insufficiency, despite a rather normal FVC%. Case presentation The patient, a 67-year-old man with non-significant past medical history, was referred to our Neurology Unit with 7-month history of progressive impairment of neck muscles that caused dropped head. Symptom onset was subtle, and it was followed by camptocormia. The neurological examination documented the camptocormic posture with a dropped head, and disclosed a mild tongue atrophy, slight hypophonia, and a mild dysphagia, with a moderate proximal and distal weakness and atrophy of the upper limbs; the lower limbs were clinically unaffected. Reflexes were present but not brisk in the four limbs. A complete diagnostic work-up according to current guidelines, including determination of serum anti-ganglioside antibodies, was performed which allowed the diagnosis of ALS. Spirometry was performed to assess the respiratory function: FVC was 80% of predicted, a value within the normal range. Diaphragm compound muscle action potentials (CMAPs) were not measured. The patient did not complain shortness of breath. Physical examination and X-ray showed no sign of diaphragmatic dysfunction. The revised ALS functional rating scale (ALSFRSR) was 42/48 [3], with a predicted intermediate rate of progression (ΔFS = 0.86). The respiratory subscores of the ALSFRS-R were 12/12. The dropped head was corrected with a collar. He started riluzole, and a 4-month follow-up visit was scheduled. Some 30 days after diagnosis, he became increasingly lethargic over a few days and was admitted to the Emergency Unit. Altered arterial blood gases were found (PaO2 26.4 mmHg, PaCO2 93.0 mmHg, pH 7.32). A CT scan of the lungs did not show radiological evidences of pneumonia or atelectasis. When inquired, relatives reported that the patient had his usual lifestyle, that he did not complain in the past days of significant respiratory problems, and that he did not take medications other than riluzole. The patient was intubated and placed on invasive mechanical ventilation, with a prompt improvement of the blood gases, i.e., PaO2 74.3 mmHg, PaCO2 46.0 mmHg, pH 7.47, and a recovery of full consciousness. The assisted mechanical ventilation was continued. One week later, the tracheal tube was removed and noninvasive ventilation (NIV) was started. In the following days,......................

We have analyzed the temporal patterns of behaviour of male rats of the Wistar and DA/Han strains on the central platform of the elevated plus maze. The ethogram encompassed 10 behavioural elements. Durations, frequencies and latencies showed quantitative differences as to walking and sniffing activities. Wistar rats displayed significantly lower latency and significantly higher durations and frequencies of walking activities. DA/Han rats showed a significant increase of sniffing duration. In addition, DA/Han rats showed a significantly higher amount of time spent in the central platform. Multivariate T-pattern analysis revealed differences in the temporal organization of behaviour of the two rat strains. DA/Han rats showed (a) higher behavioural complexity and variability and (b) a significantly higher mean number of T-patterns than Wistar rats. Taken together, T-pattern analysis of behaviour in the centre of the elevated plus maze can noticeably improve the detection of subtle features of anxiety related behaviour. We suggest that T-pattern analysis could be used as sensitive tool to test the action of anxiolytic and anxiogenic manipulations., peer-reviewed

Rationale: The largest amount of researches on the hot-plate test was carried out using quantitative assessments. However, the evaluation of the relationships among the different elements that compose the behavioral response to pain requires different approaches. Although previous studies have provided clear information on the behavioral structure of the response, no data are available on its temporal structure. Objectives: The objective of this study was to investigate the temporal structure of the behavioral response to pain in Wistar rat tested in hot-plate and how this structure was influenced by morphine-induced analgesia. Methods: The behavior of four groups of subjects tested in hot-plate, one administered saline and three with different doses (3, 6, 12mg/kg) of morphine IP, was analyzed by means of quantitative and t-pattern analyses. The latter is a multivariate technique able to detect the existence of statistically significant temporal relationships among the behavioral events in time. Results: A clear-cut influence of morphine on quantitative parameters of the response to the noxious stimulation was observed. T-pattern analysis evidenced profound structural changes of behavior. Twenty-four different t-patterns were identified following saline, whereas a dose-dependent reduction was observed following morphine. Such a reduction was accompanied by a decrease of the total amount of t-patterns detected. Conclusions: Morphine, by reducing the effects of the noxious stimulation, orients animal behavior prevalently toward exploratory t-patterns. In addition, it is suggested that the temporal structure of the response is very quickly organized and adapted to environmental noxious cues.

This study was undertaken to investigate whether, in rat interactive activities, recurring sequences of behavioural events might be identified and how and to what extent each component of the pair is involved. To this aim, the multivariate temporal-pattern (t-pattern) analysis was applied to the social interactions of 9 pairs of male Wistar rats tested in open field. Interactive activities were classified into intra- and inter-subjects. Quantitative evaluations showed that intra-subject behavioural elements represented 62.37% and inter-subject ones 37.63% of the comprehensive behaviour. T-pattern analysis revealed the presence of 221 different t-patterns organized in four different categories: containing exclusively inter-subject elements; containing both inter- and intra-subject elements; consisting of rat 1 and rat 2 intra-subject elements and, finally, consisting of intra-subject elements carried out by one of the two subjects. Results show that the activity of two interacting Wistar rats is structured on the basis of several recurring temporal sequences. Moreover, social interactions appear to be expressed also by t-patterns where the behavioural elements are carried out by animals seemingly not interacting. A support of t-pattern analysis to studies on Autism Spectrum Disorders is proposed.

The aim of the present study was to investigate structure alterations of a sandy-clay soil upon addition of different amounts of biochar (f bc ). All the f bc samples were analyzed by high energy moisture characteristic (HEMC) technique and 1H nuclear magnetic resonance (NMR) relaxometry. HEMC was applied in order to evaluate aggregate stability of biochar-amended soil samples. 1H NMR relaxometry experiments were conducted for the evaluation of the pore distributions through the investigation of water dynamics of the same samples. The HEMC technique revealed improvement in aggregate stability through measurements of the amount of drainable pores and the stability ratio. The latter increased as the amount of biochar was raised up. The 1H NMR relaxometry revealed a unimodal T 1 distribution for both the sole sandy-clay soil and the biochar. Conversely, a bimodal T 1 distribution was acquired for all the different f bc samples. Improvement in aggregate stability was obtained as biochar was progressively added to the sandy-clay soil. A dual mechanism of water retention has been hypothesized. In particular, intra-aggregate porosity was indicated as the main responsible for molecular water diffusion when f bc comprised between 0 and 0.33. Conversely, inter-aggregate porosity resulted predominant, through swelling processes, when f bc overcame 0.33.

Markers of histaminergic dysregulation were found in several neuropsychiatric disorders characterized by repetitive behaviours, thoughts and stereotypies. We analysed the effect of acute histamine depletion by means of i. c.v. injections of alpha-fluoromethylhistidine, a blocker of histidine decarboxylase, on the temporal organization of motor sequences of CD1 mice behaviour in the open-field test. An ethogram encompassing 9 behavioural components was employed. Durations and frequencies were only slightly affected by treatments. However, as revealed by multivariate t-pattern analysis, histamine depletion was associated with a striking increase in the number of behavioural patterns. We found 42 patterns of different composition occurring, on average, 520.90±50.23 times per mouse in the histamine depleted (HD) group, whereas controls showed 12 different patterns occurring on average 223.30±20.64 times. Exploratory and grooming behaviours clustered separately, and the increased pattern complexity involved exclusively exploratory patterns. To test the hypothesis of a histamine-dopamine interplay on behavioural pattern phenotype, non-sedative doses of the D2/D3 antagonist sulpiride (12.5-25-50mg/kg) were additionally administered to different groups of HD mice. Sulpiride counterbalanced the enhancement of exploratory patterns of different composition, but it did not affect the mean number of patterns at none of the doses used. Our results provide new insights on the role of histamine on repetitive behavioural sequences of freely moving mice. Histamine deficiency is correlated with a general enhancement of pattern complexity. This study supports a putative involvement of histamine in the pathophysiology of tics and related disorders.

Aim of present research was to assess, by means of descriptive and multivariate approaches, in Wistar rats tested in hole-board apparatus, whether a microstructure of hole exploratory behavior may be identified in the behavioral response to anxiety. Two groups of 15 subjects were tested: one received 1 ml of saline IP and one was IP injected with diazepam at 0.5 mg/kg dissolved in 1 ml of saline. Following saline, six not yet reported hole-exploratory components were identified: active-dip, static-dip, rapid-dip, circular-sniff, point-sniff and central-sniff. Diazepam provoked significant changes of their per cent distribution. Results of multivariate analyses revealed a complex microstructure of hole-exploratory elements characterized by behavioral flows between sniffing and dipping patterns. Diazepam significantly modified several patternings, mainly those involving point-sniff and static dip. Hence, the hypothesis of an important role of such behavioural elements and of their patternings, in anxiety-related hole-exploratory behavior, can be proposed. It is suggested that modifications of point-sniff and static-dip patternings could evidence even subtle changes of animal anxiety condition.

Aim of present paper was to carry out an analysis of rat behavior in hole-board following different multivariate approaches. Thirty male Wistar rats were observed in a hole-board apparatus and their behavior recorded for 10 min through a digital videocamera for a following frame-by-frame analysis. Both descriptive and multivariate analyses were used. Descriptive analysis showed that roughly 85% of the whole behavioral structure encompassed six patterns appearing during the first minute of observation: walking, climbing, rearing, immobile-sniffing, edge-sniff and head-dip. As to multivariate approach, cluster analysis showed three main dyadic associations: [edge-sniff/head-dip], [walking/climbing], [face-grooming/body-grooming]. Path diagram, obtained on the basis of relative frequencies of transitions among behavioral patterns (stochastic analysis), emphasized cluster analysis results. Adjusted residuals confirmed, from a statistical point of view, the strong relationships among specific patterns. Results demonstrate that immobile-sniffing has a crucial role in rat behavioral organization and head-dip is closely related with edge-sniff which is suggested to be a specific sniffing activity of holes edge never properly considered. Present research shows that multivariate approaches, revealing patterning among different behavioral elements in hole-board, could improve test reliability, providing a more useful tool to investigate behavioral effects of anxiolytic drugs.

Nicotine is one of the most addictive drugs of abuse. Tobacco smoking is a major cause of many health problems, and is the first preventable cause of death worldwide. Several findings show that nicotine exerts significant aversive as well as the well-known rewarding motivational effects. Less certain is the anatomical substrate that mediates or enables nicotine aversion. Here, we show that acute nicotine induces anxiogenic-like effects in rats at the doses investigated (0.1, 0.5, and 1.0 mg/kg, i.p.), as measured by the hole-board apparatus and manifested in behaviors such as decreased rearing and head-dipping and increased grooming. No changes in locomotor behavior were observed at any of the nicotine doses given. T-pattern analysis of the behavioral outcomes revealed a drastic reduction and disruption of complex behavioral patterns induced by all three nicotine doses, with the maximum effect for 1 mg/kg. Lesion of the lateral habenula (LHb) induced hyperlocomotion and, strikingly, reversed the nicotine-induced anxiety obtained at 1 mg/kg to an anxiolytic-like effect, as shown by T-pattern analysis. We suggest that the LHb is critically involved in emotional behavior states and in nicotine-induced anxiety, most likely through modulation of monoaminergic nuclei., peer-reviewed

Aim of present study was to investigate in male Wistar rats, whether behavioral response to hot plate test application could be influenced by systemic administration of 7-OH-DPAT, a dopaminergic (DA) D3 versus D2 receptor agonist, or U 99194, a DA D3 versus D2 receptor antagonist. Each trial lasted no more than 10 s and the whole experimental session lasted 120 min. Animal behavior was recorded by means of a digital videocamera and later, frame by frame examined using a professional videorecorder. Latency of each behavioral pattern, characterizing the response, was analysed, showing significant changes only with U 99194. A multivariate cluster analysis indicated the presence of three main behavioral clusters (exploratory, primary responses to pain, escape) which, also, resulted significantly modified by both drugs. In addition, diagrams of preferential direction, obtained through multivariate stochastic analysis, evidenced switching probabilities differences among different patterns and clusters. Results demonstrate that the behavioral response to hot plate test application is altered following 7-OH-DPAT or U 99194 administration. These findings are discussed in terms of a) drugs influence on behavioral switching and learning processes; b) a likely activity on DA D3 versus D2 receptors.

The aim of the present research was to study the effects of different basal levels of anxiety on the behavioral shift studied in the central platform of the elevated plus maze. To this purpose, quantitative and multivariate analyses, the latter based on transition matrix elaboration, were carried out on Wistar and on DA/Han rats the latter belonging to a strain characterized by different reactivity to anxiogenic stimuli. Wistar rats spent 74.11 ± 5.11 s in the central platform, whereas DA/Han significantly more: 127.08 ± 9.87. Per cent distributions evidenced a clear-cut difference in walking activities (46.25% in Wistar, 28.4% in DA/Han rats) and in the sniffing activities (45.82% in Wistar, 62.54% in DA/Han). Mean frequencies of each behavioral element showed in DA/Han strain a value significantly lower than in Wistar for central-platform entry, open arm-entry and closed-arm entry and a significant higher value for central- platform sniffing, open-arm sniffing and corner-rearing. Moreover, the ratio open-arm entry/open-arm sniffing and closed-arm entry/closed-arm-sniffing showed significant higher values in the Wistar strain. Finally, by means of hierarchical clustering analysis, strong differences between the two strains were observed in the behavioral architecture: a cascade-shaped dendrogram, branching from Walking activities, indicates that Wistar rat behavior is oriented to cross the central platform so to rapidly reach an arm; on the contrary, the dendrogram of DA/Han rats displays a behavior heavily oriented toward the permanence in the central platform. The results show that different basal levels of anxiety provoke significant differences in the behavioral shift studied in the central platform of the elevated plus maze. Such differences, evidenced by means of transition matrices elaboration, might represent the behavioral expression of anxiety-induced modifications of decision making process underlying behavioral shift activities.

The aim of the present study was to evaluate, by means of quantitative and multivariate analyses, the effects of diazepam on the behavioral structure of the rat's response to pain in the hot-plate test as well as whether such changes are associated with drug-induced effects on anxiety and/or nociception. To this purpose, ten groups of male Wistar rats were intraperitoneally injected with saline, diazepam (0.25, 0.5 and 2 mg/kg), FG-7142 (1, 4 and 8 mg/kg) or morphine (3, 6 and 12 mg/kg). The mean number and mean latency to first appearance were calculated for each behavioral component. In addition, multivariate cluster and adjusted residual analyses based on the elaboration of transition matrices were performed. Three main behavioral categories were identified: exploratory (walking, sniffing), primary noxious-evoked (hind paw licking, front paw licking, shaking/stamping) and escape (climbing, jumping). Although no significant modifications in the latencies of the primary noxious-evoked components were induced by treatment with diazepam or FG-7142, significant effects were provoked by morphine treatment. Multivariate analyses showed that diazepam-induced anxiolysis redirected the rat's behavior toward a more purposeful and effective escape strategy. In contrast, the high level of anxiety induced by FG-7142 caused the behavioral structure to become disorganized and not purposefully oriented. Changes in the organization of behavioral components were observed in morphine-treated animals and mainly consisted of modifications in the primary noxious-evoked and escape components. The findings suggest that the effects of diazepam on the structure of the rat's response to pain in the hot-plate test are more likely attributable to anxiolysis than pain modulation.

Aim of the research was to study, by means of descriptive and multivariate analyses, whether, and how, learning influences the behavioral structure of rat response to pain. To this purpose, a hot-plate test daily repetition procedure was carried out on male Wistar rats for five days. A 6-day interval without stimulation elapsed before last test was carried out on day 12. After composition of an ethogram, descriptive (number, latency, per cent distribution) and multivariate analyses (cluster, stochastic) were carried out for each scheduled test day. One-way ANOVA and Newman-Keuls post-hoc test for multiple comparisons revealed significant changes for climbing, jumping, front-paw licking and stamping mean latencies. Student's t-test, carried out between days 1 and 12, showed that significant modifications persisted over time only for climbing and jumping. Cluster analysis evidenced three different clusters: exploratory (sniffing, walking), primary noxious evoked elements (front-paw licking, hind-paw licking, stamping), and escape (climbing and jumping), each showing modifications during test repetition. Comparison between days 1 and 12 revealed substantial differences in similarity values of escape patterns whereas, for the response to pain ones, a conservative structure of dendrograms was maintained. Stochastic analysis revealed a progressive increase of transitions toward jumping and a significant reduction of the ones between sniffing and walking. Such modifications persisted also after 6 days without any stimulation. Present study shows that learning provokes a complex and fine temporal evolution of the innermost behavioral structure of rat response to pain aiming at a more efficient escape strategy.

The aim of the present methodological article was to assess whether a microstructure of hole-exploratory activity can be demonstrated in rats tested in the hole-board, an experimental assay widely used to study anxiety-related behaviors in rodents. Six never properly described hole exploratory behavioral components were identified: active-dip, static-dip, rapid-dip, circular-sniff, point-sniff and central-sniff. Results, assessed by means of descriptive and multivariate approaches, reveal a complex microstructural organization of these behavioral elements. The hypothesis of an important role of the relationships between point-sniff and head dipping activities, in anxiety-related hole exploratory behavior, is proposed. It is suggested that the microstructural analysis can evidence even subtle behavioral changes induced by manipulations of animal anxiety condition.

Rationale: By means of t-pattern analysis, it has been observed that the different events, characterizing rat behavior in hole board (HB), present close interrelationships which occur sequentially and with significant constraints on the interval lengths separating them. Objectives: The aim of present research was to study, by means of descriptive and multivariate t-pattern analyses, the effects of the reference anxiolytic drug diazepam (DZP) on temporal structure of a rat’s anxiety-related behavior in HB. Methods: Fifty-six male Wistar rats were tested for 10 min in HB. Video files, collected for each animal, were coded by means of a software coder, and event log files, generated for each subject, were analyzed by means of a specific software for temporal pattern analysis (t-pattern analysis). Results: Significant diazepam-induced modifications were observed for durations of walking, climbing, edge-sniff, and face grooming. Dose-dependent decreases of t-patterns’ total amount, of their mean occurrences and of their mean length for each group were detected. Also, t-patterns’ mean occurrences, in terms of different composition, were reduced. Percent distributions showed a significant increase of t-patterns including walking for all administered groups, and significant reductions of t-patterns including climbing, immobile sniffing, and edge-sniff. Front-paw licking and face grooming were reduced at the higher DZP dose. Conclusions: Present study demonstrates, for the first time, that the temporal structure of Wistar rats’ behavioral response to anxiety in HB changes following pharmacological manipulation of anxiety condition. Moreover, t-pattern analysis is suggested to represent a useful tool to evaluate and compare different classes of anti-anxiety molecules.

In our previous study we suggested that multivariate analysis could improve hole-board test reliability providing a more useful tool to determine behavioral effects of anxiolytic drugs. To support this hypothesis, a multivariate analysis of rat behavior in hole-board, following administration of the reference anxiolytic drug diazepam, was carried out. Four groups, each composed of thirty male Wistar rats, were used: one saline and three diazepam injected (0.25, 0.5 and 2 mg/kg IP). Rat behavior was recorded for 10 min through a digital videocamera. Descriptive and multivariate analyses were carried out. In all groups, more than 80% of whole behavioral structure encompassed walking, climbing, rearing, immobile-sniffing, edge-sniff and head-dip. Moreover, modifications observed of a specific index, represented by edge-sniff/head-dip ratio, were correlated to diazepam-induced modifications of anxiety level. Cluster analysis showed that diazepam at 0.5 and 2 mg/kg induced important changes for [edge-sniff/head-dip] cluster. In addition, in all diazepam groups a [walking/climbing] cluster appeared. Path diagrams showed close relationships among different patterns both in saline and diazepam injected animals. Also, significant changes were detected following diazepam for transitions encompassing both general exploratory patterns (walking, climbing) and the specific ones (head-dip and edge-sniff). Adjusted residuals confirmed in all groups patterns relationships and, where present, significant behavioral associations. Results demonstrate that an anxiolytic activity can be revealed by head-dip/edge-sniff association weakening and by the addressing of behavioral structure toward general exploratory activity. Improvement of hole-board test reliability in behavioral study of anxiety, following multivariate analysis, is emphasized.

6-Hydroxydopamine (6-OHDA) is a neurotoxic compound commonly used to induce dopamine (DA) depletion in the nigrostriatal system, mimicking Parkinson's disease (PD) in animals. The aim of the present study was to evaluate the 7-day effect of unilateral nigral lesion on rat brain monoamine neurochemistry. Five brain regions were examined: the brain stem, cerebellum, hippocampus, striatum, and cortex. 6-OHDA-unilateral lesion dramatically modified DA, serotonin (5-HT) and their metabolites contents in both sides of the different brain nuclei. Furthermore, unilateral 6-OHDA lesion reduced DA and 5-HT contents and produced a robust inversion of their turnover in the nonlesioned side compared to sham-operated rats. These data suggest that 6-OHDA unilateral nigral lesion produces bilateral monoamine level modifications, and this piece of evidence should be taken into account when one interprets data from animal models of unilateral PD., peer-reviewed

Nitric oxide (NO) signalling plays an important role in the integration of information processed by the basal ganglia nuclei. Accordingly, considerable evidence has emerged indicating a role for NO in pathophysiological conditions such as Parkinson’s disease (PD), schizophrenia and drug addiction. To further investigate the NO modulation of dopaminergic function in the basal ganglia circuitry, in this study we used in vivo electrophysiology and microdialysis in freely-moving rats. Pharmacological manipulation of the NO system did not cause any significant changes either in the basal firing rate and bursting activity of the dopamine (DA) neurons in the substantia nigra pars compacta (SNc) or in DA release in the striatum. In contrast, the disruption of endogenous NO tone was able to counteract the phasic dopaminergic activation induced by nicotine treatment in both experimental approaches. These results further support the possibility that nicotine acts via a NO mechanism and suggest a possible state-dependent facilitatory control of NO on the nigrostriatal DA pathway. Thus, NO selectively modulates the DA exocytosis associated with increased DA functiov n., peer-reviewed

The present study was undertaken to explore the involvement of nitric oxide (NO) in the 6-hydroxydopamine (6-OHDA) experimental model of Parkinson's disease (PD) in rats. The effect of pharmacological manipulation of the NO system was evaluated on striatal dopamine (DA) level decrease produced by the toxin. 7-nitroindazole (7-NI, 50 mg/kg i.p.; n= 5) pretreatment significantly restored the striatal DA contents. Conversely, 40 mg/kg i.p. of molsidomine (MOL, n= 5), an NO donor, significantly worsened the neurodegeneration (n= 5) and completely counteracted the neuroprotective effect of 7-NI (n= 5). Thus, a crucial role for NO in 6-OHDA induced neurodegeneration is suggested together with a protective benefit for inhibitors of NOS in the treatment of PD.

Parkinson's disease (PD) is known to be a chronic and progressive neurodegenerative disease caused by a selective degeneration of dopaminergic (DAergic) neurons in the substantia nigra pars compacta (SNc). A large body of experimental evidence indicates that the factors involved in the pathogenesis of this disease are several, occurring inside and outside the DAergic neuron. Recently, the role of the neuron–glia interaction and the inflammatory process, in particular, has been the object of intense study by the research community. It seems to represent a new therapeutic approach opportunity for this neurological disorder. Indeed, it has been demonstrated that the cyclooxygenase type 2 (COX-2) is up-regulated in SNc DAergic neurons in both PD patients and animal models of PD and, furthermore, non-steroidal anti-inflammatory drugs (NSAIDs) pre-treatment protects against 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) or 6 hydroxydopamine (6-OHDA)-induced nigro-striatal dopamine degeneration. Moreover, recent epidemiological studies have revealed that the risk of developing PD is reduced in humans who make therapeutical use of NSAIDs. Consequently, it is hypothesized that they might delay or prevent the onset of PD. However, whether or not these common drugs may also be of benefit to those individuals who already have Parkinson's disease has not as yet been shown. In this paper, evidence relating to the protective effects of aspirin or other NSAIDs on DAergic neurons in animal models of Parkinson's disease will be discussed. In addition, the pharmacological mechanisms by which these molecules can exert their neuroprotective effects will be reviewed. Finally, epidemiological data exploring the effectiveness of NSAIDs in the prevention of PD and their possible use as adjuvants in the therapy of this neurodegenerative disease will also be examined., peer-reviewed

Drugs that selectively block D 2 receptors are known to provoke a rapid cell firing increase followed by A9 and A10 dopaminergic (DA) neuron inactivation (depolarization block). In this study, possible relationships between cell firing rapid increase and specific behavioral effects, linked to sensorimotor integration, were investigated in the rat. To this purpose, with the aid of a video camera apparatus and a frame-by-frame analysis, effects of sulpiride-induced blockade of DA D 2 receptors were analyzed on the orienting movement of the head induced by acoustic stimulation. In a control group of rats, during trials lasting 20 min, latency and duration of head turning (HT) were 186.15 ± 51.66 ms and 266.87 ± 47.49 ms, respectively. Sulpiride injection (20,40,100 mg/kg IP) provoked a dose-dependent increase of HT latency and duration; however, only latencies showed statistically significant variations. It is suggested that cell firing rapid increase, observed on A9 and A10 DA neurons, following sulpiride administration, may be correlated to rapid modifications of specific HT parameters linked to sensorimotor integration.