Recent work has advanced our knowledge of phasic dopamine reward prediction error signals. The error signal is bidirectional, reflects well the higher order prediction error described by temporal difference learning models, is compatible with model-free and model-based reinforcement learning, reports the subjective rather than physical reward value during temporal discounting and reflects subjective stimulus perception rather than physical stimulus aspects. Dopamine activations are primarily driven by reward, and to some extent risk, whereas punishment and salience have only limited activating effects when appropriate controls are respected. The signal is homogeneous in terms of time course but heterogeneous in many other aspects. It is essential for synaptic plasticity and a range of behavioural learning situations. Current Opinion in Neurobiology 2013, 23:229–238MacrocircuitsThis review comes from a themed issue on Steve Petersen and Wolf SingerEdited by Issue and the EditorialFor a complete overview see the Available online 22nd December 20120959-4388/$ – see front matter, © 2012 Elsevier Ltd. All rights reserved.http://dx.doi.org/10.1016/j.conb.2012.11.012The smoke signals of the American Indians and the peep-peep from Sputnik demonstrate that information processing systems use signals. The brain is no exception. Its action potentials are instrumental for extracting information from the environment and directing behaviour. The foremost function of the brain is to assure individual survival and gene propagation for ultimate evolutionary fitness. To this end individuals need to acquire specific substances for their bodily functions. The substances come in foods and liquids and require effort to obtain. They are called rewards and support learning, approach behaviour, decision making and positive emotions like pleasure, desire and happiness. Numerous other objects, events and situations have similar reward functions and contribute to evolutionary fitness. To understand reward processing, we need to study neuronal signals for these objects. We can describe reward functions in formal behavioural terms like Pavlovian and operant conditioning, habits and goal directed behaviour, economic decision making and see how alterations in brain functions influence these processes. But to understand how the brain as an information processing machine mediates reward we need to take these terms apart and identify and understand neuronal signals for their components. This review provides an update on dopamine neurons that provide a reward signal for updating economic decision variables.Most midbrain dopamine neurons (75–80%) show rather stereotyped, phasic activations with latencies of <100 ms and durations of <200 ms following unpredicted food or liquid rewards. This response codes a quantitative prediction error, namely the difference between received and predicted reward value. A reward that is better than predicted elicits an activation (positive prediction error response), a fully predicted reward draws no response, and a reward that is worse than predicted induces a depression (negative error response). Most dopamine neurons (60–75%) respond in similar ways to reward predicting stimuli, coding positive and negative, higher order reward prediction errors at the time of the stimulus relative to the prediction at that moment •,4••.The positive prediction error response can amount to phasic tripling of impulse rate, whereas the negative response has limited dynamic range owing to the low baseline activity of dopamine neurons (3–5 impulses/s), which might suggest limited negative coding . HoweverThe dopamine response to reward is compatible with the notion of primary reinforcer prediction error according to error driven learning rules . These lReinforcement learning establishes reward predictions that are essential for informed decision making . Model-fIn learning situations governed only by experienced rewards, consecutive unrewarded trials lead to progressively decreasing reward prediction. Correspondingly, positive prediction errors to an unpredicted reward increase, because the repeated absence of reward makes the reward less expected and the delivery of reward becomes more surprising. However, in particular tasks, sequences of unrewarded trials may lead to increasing reward probability (increasing hazard rate) and thus increasing reward prediction. Now the reward becomes progressively less surprising and hence induces a prediction error that decreases over successive trials. Learning in such a task benefits from knowledge about the underlying model of the world. Indeed, dopamine error responses follow precisely this scheme, decreasing with increasing numbers of trials since the last reward , suggestDopamine responses are influenced by previously acquired information about reward probability distributions . The errRepeated reversals of stimulus-reward or position-reward associations lead to acquisition of a rule that helps individuals reverse their reactions within one trial . Correspondingly, dopamine responses to reward predicting stimuli reverse after one trial without having experienced the contiguity between the newly valid stimulus position and the reward ••. TheseIn each of these examples, the observed dopamine prediction error responses are not fully explained by the actual experience with reward but incorporate information from models of the world. Thus the dopamine prediction error responses occur in both model-free and model-based reinforcement learning situations, without evidence so far for a role in initial model acquisition.Individuals make choices that maximise their rewards. Although rewards have objective, physical and chemical properties, their value is defined by the needs of the individual decision maker and thus intrinsically subjective. One way to distinguish subjective from objective reward value is temporal discounting, the decrease in subjective reward value with increasing delays despite constant physical size. Psychometric analysis of choices in monkeys with variable early vs. constant 4, 8 and 16 s delayed rewards reveals graded hyperbolic decays in subjective value by 25, 50 and 75%, respectively, compared to reward at 2 s . ExponenDopamine responses to reward predicting stimuli decrease monotonically across reward delays of 2–16 s in monkeys, despite the same physical amount of reward being delivered after each delay  Figure b. ResponReward neurons in most other primary and associated reward structures of the brain show some degrees of temporal discounting, including orbitofrontal cortex , prefronIf dopamine neurons code reward value subjectively in temporal discounting, would they show other aspects of subjective coding compatible with the notion of an intrinsically subjective survival function of rewards? For instance, would their reward prediction coding reflect the physical presence or the subjective experience of predictive stimuli? Signal detection theory offers well proven tests. Monkeys correctly or incorrectly report the presence of stimuli , and correctly or incorrectly report stimulus absence  ••. DopamThe relationship to subjective perception is also seen with dopamine error responses to subsequent cues and reward. Correct and incorrect reports of stimulus presence lead to reward in 76% of trials, whereas correct and incorrect reports of stimulus absence lead to 64% reward ••. As doTogether with the responses to the near threshold stimulus itself and the subjective value coding in temporal discounting, dopamine responses reflect the animal's subjective perception and valuation of the stimuli beyond purely physical reward properties. As decisions are ultimately made according to subjective reward values, the dopamine responses may provide rather direct and parsimonious, and therefore evolutionary beneficial and selected, inputs to neuronal decision processes.Electric shocks, painful pinches and airpuffs, as well as conditioned stimuli predicting these events, induce phasic depressions in dopamine impulse activity in anaesthetised monkeys and rodents • and in A few dopamine neurons (5–15%) are activated by primary aversive stimuli , which iin vivo dialysis over seconds [Aversive stimuli increase striatal dopamine concentrations measured by  seconds . The inc seconds  and impu seconds •,45. Fas seconds  and incr seconds . The inc seconds , some ofSubstantial fractions of dopamine neurons (35–65%) are activated by conditioned aversive stimuli presented in random or blockwise alternation with reward predicting stimuli , which eThe initial, brief activation is identical for all conditioned reward predicting, unrewarded, differently valued and aversive stimuli, with <100 ms latency and 50–150 ms duration . It is sThe second response component constitutes a genuine value response  which diThe two response components are better separated in a dot motion task that requires time for stimulus identification • Figuref. The inTaken together, the more frequent activations to conditioned aversive stimuli compared to primary punishers is probably explained by the nature of the two response components. The initial, stereotyped activation constitutes a pre-identification, higher order temporal TD error response, or possibly response generalisation to rewarding stimuli or contexts. After proper identification of the stimulus, the second component codes a TD error accurately. A fast but inaccurate initial neuronal response may provide survival advantages. It could lead to early initiation of behavioural reactions in the course of which the exact nature of the stimulus becomes identified by the second component. The mechanism would allow the agent to arrive first at the object and thus provide a slight advantage over slower competitors, which would ultimately pan out as higher evolutionary fitness. If the initial response is false and the object unrewarded, behavioural reactions can be cancelled while the second response component identifies the object 200 ms later, which would be a small price to pay.Whereas interest in dopamine reward responses has overshadowed the long known aversive responses, some current work addresses punishment in identifiable groups of dopamine neurons without assessing their incidence relative to reward responsive neurons . AlthougSalience refers to the capacity of stimuli to elicit arousal, alert and attention which enhance neuronal processing and behavioural reactions. There are, in principle, three forms of salience. Physical salience derives from physically strong stimuli. Novelty or surprise salience originates in novel or surprising events. Motivational salience is produced commonly by rewards and punishers. Would salience explain the higher incidence of responses to conditioned aversive stimuli (35–65%) compared to unpredicted air puffs (<15%) ? The phyFurther arguments limit motivational salience as a major factor determining dopamine responses. First, the change from visual to auditory rewarded stimuli reduces aversive dopamine activations to stimuli without reducing the salience of the unchanged aversive stimulus . Second,Whereas 70–80% of dopamine neurons code reward prediction errors, only 10–15% are activated by both rewarding and aversive events and thus motivational salience. Therefore motivational salience induces 5–8 times less dopamine activations than reward, rather than similar proportions as suggested . The depTogether with the previously described risk responses in 30% of dopamine neurons , dopaminThe reward responses are homogeneous in terms of latency, duration, positive monotonicity of value coding and error coding  and someThe bidirectional dopamine prediction error response at the time of the reward implements fully the error term of the Rescorla-Wagner learning rule . The dopA viable neuronal reinforcement mechanism involves three-factor Hebbian plasticity with dopamine and glutamate synapses located at the same dendritic spines of striatal or frontal cortical neurons, the so-called synaptic triad . Indeed,Protocols for spike time dependent plasticity (STDP) demonstrate LTP when presynaptic stimulation precedes postsynaptic stimulation by a few tens of milliseconds, whereas LTD occurs with reverse sequence. Intact dopamine D1 receptors are required for both forms of cortically evoked plasticity in striatal neurons involving NMDA receptors ••. When Dopamine neurons show induction of NMDA receptor dependent LTP when a short postsynaptic burst occurs after at least 500 ms of presynaptic stimulation •. A delaDopamine exerts immediate postsynaptic effects during behavioural performance and approach behaviour. At striatal neurons of the direct pathway, dopamine has excitatory effects via the D1 receptor by eliciting or prolonging glutamate inputs and transitions to the up state (depolarisation) of the membrane potential, whereas in indirect pathway neurons D2 receptor activation has inhibitory effects by reducing glutamate release and prolonging membrane down states (hyperpolarisation) . These iHundreds of lesioning and psychopharmacological studies using various tasks demonstrate learning deficits with impaired dopamine transmission. At the level of dopamine neurons, knock outs of burst generating NMDA receptors in mice, with resulting decreases in responses to reward predicting stimuli, lead to a range of learning deficits, including conditioned place preference, maze learning and operant conditioning, that are dissociated from performance deficits •,85 Figc, d. In By contrast, learning is less impaired by systemic D1 receptor blockade in select tasks with separate stimulus and goal locations that fail to engage dopamine neurons •• or wheTaken together, learning depends on intact dopamine function in simple reward contiguity situations with explicit, easily identifiable rewards and conditioned stimuli that engage phasic dopamine responses. Learning may not depend on dopamine neurons if their phasic responses remain unengaged during learning. Despite this obvious conclusion, learning of tasks normally engaging dopamine responses might even proceed despite lesions when neuronal plasticity allows other learning systems to compensate within the tested time frame. Knock out of specific learning systems may lead to modified ontogenesis with even more opportunity for shifts to other systems. Given the crucial importance of reward for survival, multiple, flexible learning systems are biologically plausible and would enhance evolutionary fitness.• of special interest•• of outstanding interestPapers of particular interest, published within the period of review, have been highlighted as:


Referring to Europeans as Caucasians, as still regularly done in the medical and less frequently in the population genetic literature, most often is not justified according to modern scientific views. This term, first mentioned in 1785 by Christoph Meiners, was picked up by Johann Friedrich Blumenbach in his 1795 suggestion of dividing the human world population into 5 groups . Nevertheless, a Caucasian classification can still be formally correct if it refers to people who originate from the Caucasus region. One real Caucasian, the molecular anthropologist Dr. Ivan (Vano) Nasidze , he dedicated almost all of his professional life to the genetic investigation of human population relationships and history in the Caucasus region. His latest (and now last) in a series of articles on this topic was published in September last year in the  Genetics. Using cI will miss Vano as a friendly and cooperative colleague who was deeply committed to explore the genetic history of the Caucasus peoples with various molecular approaches . I sincerely hope that Dr. Ivan Nasidze will be remembered for his important contributions to the investigative genetics and the molecular anthropology of the Caucasus for a long time to come.


Synchronized oscillations between neurons or across nuclei within parkinsonian basal ganglia (BG) have been extensively reported over past decade. They are implied in the hypokinetic motor symptoms of Parkinson’s disease . Thus, iDopamine loss, a characteristic feature of Parkinson’s disease generally accepted to cause motor symptoms, is known to affect network properties as well cellular properties of STN-GPe network. In this study, we considered how these changes due to dopamine loss contribute to the generation of pathological rhythms in STN-GPe network. We reduced large STN-GPe network model  (each nuThere is an experimental evidence for how dopamine presynaptically suppresses different types of basal ganglia synapses. We show how the chance in strength of dopamine-modulated synapses promotes intermittent synchronous activity patterns under very general conditions and investigate the cellular properties, necessary for this dynamics. It is also known that dopamine affects not only synaptic properties, but also cellular properties of BG neurons. In particular, change in the dopaminergic status leads to change in the temporal features of firing patterns of STN neuron. So we also considered how dopamine-dependent features of STN neuron firing modulate the GPe response and the synchrony between STN input and GPe output. As STN dopaminergic modulation and STN activity pattern vary, we observe the overall transition from low coherence state to high coherence state through intermittent synchronous activity. This points to the important role of GPe cells within BG circuit and demonstrate that the response of GPe cell, which was achieved through the interaction between the intrinsic properties of the cell itself and membrane properties of STN cell, provide good source for the temporal characteristics of synchrony observed in experimental setting.In sum, our modeling studies explain how the lack of dopamine in Parkinson’s disease may lead to the generation of intermittent synchronous dynamics in the basal ganglia via two different pathways: cellular and synaptic.


Eosinophilic granuloma of bone is a disease with an incidence of one new case per 350,000 to 2 million per year, which is an uncommon disease of maxillofacial region, and presents in more than 90% in children under the age of ten with predominance for males. As a result, eosinophilic granuloma of the jaw is always unconsidered in the differential diagnosis of similar lesions by many clinicians. It is difficult to make a correct diagnosis on it without proof of a pathological diagnosis, which correlates with the diverse clinical and radiographic presentations of eosinophilic granuloma in the jaws. In the present paper we report a rare case of unifocal eosinophilic granuloma of mandible occurring in an adult female. The term “Eosinophilic Granuloma of bone” was introduced by Lichtenstein and Jaffe in 1940 . It is aEosinophilic granuloma may not present physical signs or symptoms in the clinical observation and most of the times it is discovered during routine radiographic examination. Sometimes there may be localized swelling, paing or tenderness. The lesion may occur in the jaw and overlying soft tissues of the mouth although the skull and mandible are common regions of involvement, the femur, ribs, humerus, and other bones may also be affected. Loss of superficial alveolar bone and localized periodontitis are common early forms of the disease. unifocal eosinophilic granuloma of bone has a destructive nature, and is well demarcated, roughly round or oval in shape. The area destroyed replaced by a soft tissue (brown in color) since there is no necrosis, later the lesion becomes fibrous and grayish.Under the direction of the Writing Group of the Histiocyte Society, Langerhans cell histiocytosis has been adopted as the appropriate clinicopathologic designation that encompasses and essentially replaces the previous historical terms used to classify this category of abnormal histiocyte proliferation . HoweverA 35-year-old female reported to the out-patient department, Career Post Graduate Institute of Dental Sciences & Hospital, Lucknow on January 3rd, 2011 with a complain of unhealed dental socket present on the right-posterior mandibular alveolar ridge. 6 weeks agothe patient visited a local dentist with complain of loose teeth, diagnosis of periapical pathology was made and she got her few teeth removed. Antibiotics and anti-inflammatory drugs where prescribed for 5 days. After 2 weeks the patient started experiencing dull continuous pain in the same region with salty taste sensation and foul smelling odor from mouth, for which she again went to the same dentist, previously prescribed drugs were repeated and betadin mouthwash was also recommended. The patient found no improvement so she decided to consult our institute. The physical and neurological examination had no pathological signs. No fever, elevated regional temperature, or lymphadenopathy were recorded with no history of diabetes,numbness, or weakness. Intraoral examination revealed unhealed sockets in respect to 46, 47 and 48. No hemorrhagic areas or purpura was observed. The differential diagnosis include osteomyelitis, Paccioni granulations, Ewing's sarcoma, osteosarcoma, multiple, myeloma and metastasis. Orthopantomograph demonstrated round/ovoid punched out lesion with beveled edge = hole-within-a-hole appearance extending from premolar area up to the right condyle. Its borders were irregular and no calcification or ossification in the lesion was noted . In fullBlood examination revealed no pathological findings, neither of elevation of leukocytes nor of erythrocyte sedimentation rate. Heart ultrasound was made which revealed no pathologic findings.Surgical curettage was done and bone graft was placed under local anesthesia and samples were sent to the department of oral pathology and microbiology for the histopathological diagnosis. Macroscopically, the gross has small, multiple masses of both hard and soft tissue measuring less than 1 cm. Soft tissue masses were granular and gelatinous, appearing gray red to brown with flecks of yellow, while bony masses appeared yellow-tan with ill-defined borders . MicroscPostoperative outcome of lesion was uneventful and showed good prognosis. After 20 weeks removable prosthesis was given and no recurrence was observed till date.Eosinophilic granuloma usually presented as a monostotic lesion affecting flat and long bones 70%) , the sku0% , the On plain radiographs, eosinophilic granuloma typically presents as a punched out lesion with reactive sclerosis. A less common finding is one of a permeative pattern with or without periosteal reaction. The calvaria and especially the parietal bones are most often affected, followed by the mandible, the ribs, and the pelvis. Lesions in the long bones are most often in the diaphysis (58%). Epiphysis is rarely involved (2%). Lesions in the calvaria are between 1 to 15 cm in diameter. In patients with systemic disease, lesions are generally larger and can measure up to 25 cm . The tumor material is sterile but there have been reports about the presence of staphylococcus and streptococcus . EosinopEosinophilic granuloma does not lead to malignant transformation. If it expands elsewhere but bones then is called Hand-Schuller-Christian disease and may manifest as diabetes insipidus, cerebellar, hypothalamic, and with other central nervous system symptoms. The prognosis depends on the age of diagnosis and the number of foci.In general, no treatment is needed for localized osseous eosinophilic granuloma and often the biopsy is enough to initiate healing. Steroid injection, curettage, excision, or radiation may be necessary depending on the extent of the disease and the symptoms.This is one of the rare cases of eosinophilic granuloma of the mandible developing in an adult female patient. It is important to include eosinophilic granuloma in the differential diagnosis of bone lesions in adult subjects because of the possible expansion of the disease if untreated. We suggest a followup of a year is necessary for probable recurrence.


Recently, we demonstrated that the central ghrelin signalling system, involving the ghrelin receptor (GHS-R1A), is important for alcohol reinforcement. Ghrelin targets a key mesolimbic circuit involved in natural as well as drug-induced reinforcement, that includes a dopamine projection from the ventral tegmental area (VTA) to the nucleus accumbens. The aim of the present study was to determine whether it is possible to suppress ghrelin's effects on this mesolimbic dopaminergic pathway can be suppressed, by interrupting afferent inputs to the VTA dopaminergic cells, as shown previously for cholinergic afferents. Thus, the effects of pharmacological suppression of glutamatergic, orexin A and opioid neurotransmitter systems on ghrelin-induced activation of the mesolimbic dopamine system were investigated. We found in the present study that ghrelin-induced locomotor stimulation was attenuated by VTA administration of the N-methyl-D-aspartic acid receptor antagonist (AP5) but not by VTA administration of an orexin A receptor antagonist (SB334867) or by peripheral administration of an opioid receptor antagonist . Intra-VTA administration of AP5 also suppressed the ghrelin-induced dopamine release in the nucleus accumbens. Finally the effects of peripheral ghrelin on locomotor stimulation and accumbal dopamine release were blocked by intra-VTA administration of a GHS-R1A antagonist (BIM28163), indicating that GHS-R1A signalling within the VTA is required for the ghrelin-induced activation of the mesolimbic dopamine system. Given the clinical knowledge that hyperghrelinemia is associated with addictive behaviours  our finding highlights a potential therapeutic strategy involving glutamatergic control of ghrelin action at the level of the mesolimbic dopamine system. Ghrelin, a gastric hormone, exerts orexigenic and pro-obesity effects by interacting with key brain circuits involved in appetite and energy balance , 2008. TPeripherally injected ghrelin also stimulates the mesolimic dopamine system . Previouam). Tap water and food  were supplied ad libitum, except during the experiments. All experiments were conducted during the day time when the mice are less active. Studies were approved by the Ethics Committee for Animal Experiments in Gothenburg, Sweden.Adult post-pubertal age-matched male NMRI mice  were used for studies of locomotor activity and dopamine release as such studies are well-documented in this strain  was diluted in 0.9% sodium chloride  and was administrated intraperitoneally (i.p.) (10 ml/kg body weight). The selected dose, 0.33 mg/kg, was determined previously, as it increases locomotor activity and accumbal dopamine release as well as induces a conditioned place preference in mice . Ghrelin2 1.2 mM; KCl 3.0 mM and MgCl2 1.0 mM)  and was administered at a dose of 2.5 µg/side  at 40 minutes prior to i.p. ghrelin/vehicle exposure. Previous studies have established that this compound is a GHS-R1A antagonist and fully inhibits ghrelin-induced GHS-R1A activation , a GHS-R1A antagonist, has also been determined previously , an N-methyl-D-aspartic acid (NMDA) receptor antagonist, was determined in a dose-response study where 0.5 µg/side  was the highest dose not to affect locomotor activity per se . AP5 or per se .per se (data not shown). Doses in a similar range have previously been shown to block the cue-induced reinstatement of cocaine seeking , an orexin A receptor antagonist, was determined in a dose-response study where 5 µg/side  was the highest dose not to affect locomotor activity  seeking . SB33486Naltrexone, an unselective opioid receptor antagonist with some selectivity to the µ receptor, was diluted in saline vehicle. Naltrexone  or saline vehicle were injected 30 minutes prior to i.p. ghrelin/vehicle. The dose was determined from previous studies in which doses in a similar range have been shown to block the reinforcing properties of alcohol in rodents . The ratIntra-VTA injections were made using a volume of 0.5 µl/side via chronically implanted catheters, over 60 seconds. A 5 µl syringe  was used to facilitate drug administration into the VTA. After each injection, the cannula was left in place for a further 60 seconds to facilitate diffusion. Potentially, the volume administered during intra-VTA injections could raise concerns about specificity due to possible leakage into neighboring structures. Previously, however, we found that only ‘on-target’ placements (in the VTA and LDTg but not in closely adjacent sites) resulted in significant effects of ghrelin or GHS-R1A antagonists on locomotor stimulation and dopamine release using a larger volume (1 µl) for mice . SupportPeripheral administration of ghrelin has previously been shown to stimulate locomotor activity in mice . LocomotLocomotor activity was registered in eight sound attenuated, ventilated and dim lit locomotor boxes . Five by five rows of photocell beams, at the floor level of the box, creating photocell detection allowed a computer-based system to register the activity of the mice. Locomotor activity was defined as the accumulated number of new photocell beams interrupted during a 60-minute period.Before initiating the experiments, a dummy cannula was carefully inserted and retracted into the guide cannula to remove clotted blood and hamper the spreading depression. Mice were then allowed to habituate to the locomotor activity box one hour prior to drug challenge. In separte experiments, the effects of i.p. administered ghrelin on locomotor stimulation was investigated following intra-VTA administration of BIM28163, AP5 or SB334867 to mice. In subsequent experiments, naltrexone was injected i.p. prior to ghrelin. In the first locomotor activity experiment, BIM28163 (2.5 µg/side) or an equal volume (0.5 µl/side) of vehicle solution (Ringer) was administered locally and bilaterally into the VTA. Ghrelin (0.33 mg/kg) or an equal volume of vehicle solution  was thereafter injected. The same experimental protocol was used for AP5 (0.5 µg/side), SB334867 (5 µg/side) and naltrexone . All mice received drug treatment only twice (antagonist/vehicle and ghrelin/vehicle). Neither water nor food was available to the mice during the locomotor experiments. The activity registration started five minutes after the last injection and was subsequently measured for a 60-minute period. For intra-VTA administration only mice with guide cannulae placements in the VTA were included in the statistical analysis.Locomotor activity experiments). The coordinates for the N.Acc. shell were: 1.5 mm anterior to the bregma, ± 0.7 lateral to the midline and 4.7 mm below the surface of the brain and the coordinates for the VTA were 3.4 posterior to bregma, ±0.5 mm lateral to the midline and 1.0 mm below the brain surface (For measurements of extracellular dopamine levels and overflow (that reflect dopamine release), mice were implanted unilaterally with a microdialysis probe positioned in the N.Acc. shell and a guide cannulae into the VTA. The probe and the guide cannula/e were positioned ipsilateral, and the location was randomly alternated to either the left or right side. The surgery was preformed two days prior to the experimental day as described above  and perfused with Ringer solution at a rate of 1.5 µl/minute. After one hour of habituation to the microdialysis set-up, perfusion samples were collected every 20 minutes. The baseline dopamine level was defined as the average of three consecutive samples before the first drug/vehicle challenge. After the baseline samples, the antagonist (AP5 or BIM28163) was administered locally into the VTA followed subsequently by a ghrelin (i.p.) injection. The dopamine levels in the dialysates were determined by HPLC with electrochemical detection. A pump , an ion exchange column  and a detector  equipped with a VT-03 flow cell (Antec Leyden) were used. The mobile phase (pH 5.6), consisting of sulfonic acid 10 mM, citric acid 200 mM, sodium citrate 200 mM, 10% EDTA, 30% MeOH, was vacuum filtered using a 0.2 µm membrane filter . The mobile phase was delivered at a flow rate of 0.2 ml/minute passing a degasser , and the analyte was oxidized at +0.4 V.After completion of the microdialysis experiments, the locations of the probe and guide cannulae were verified. Neither water nor food were available to the mice during the microdialysis experiment. Only mice with probe placement in the N.Acc. and guide cannulae in the VTA were included in the statistical analysis.After the locomotor activity and microdialysis experiments were completed, the location of the probe and/or cannula/e were verified. The mice were decapitated, probes were perfused with pontamine sky blue 6BX to facilitate probe localization, and the brains were mounted on a vibroslice device . The brains were cut in 50 µm sections and the location of the probe and/or cannula/e was determined by gross observation using light microscopy. The exact position (some correct and some misplaced) of the probe and/or guide cannula/e was verified .P < 0.05 was considered as statistically significant.All locomotor activity data were evaluated by a two-way ANOVA followed by Tukey's HSD post-hoc tests comparing treatments. The microdialysis experiments were evaluated by a two-way ANOVA for repeated measures followed by Tukey's HSD post-hoc test for comparisons between different treatments and specifically at given time points. Data are presented as mean ± SEM. A probability value of per se (P < 0.01) was attenuated by VTA administration of BIM28163 (P < 0.01) in mice  = 5.45, P = 0.005: n = 6–8). In the microdialysis experiments a significant effect of systemic ghrelin to increase dopamine release in comparison to vehicle treatment was observed (P = 0.003). Pre-treatment with BIM28163 attenuated the ghrelin-induced increase in dopamine release compared with vehicle pre-treatment in mice (P = 0.001)  = 6.39, P = 0.002; time F = 1.77, P = 0.047; treatment-time interaction F = 4.01, P < 0.001). This difference was evident at the time intervals 20–100 minutes (P < 0.001: n = 7–8).First, the role of GHS-R1A receptors in the VTA for the reinforcing effects of ghrelin by tests of ghrelin-induced locomotor stimulation and, in separate studies, by measurement of ghrelin-induced dopamine release were investigated. The locomotor stimulatory and accumbal dopamine releasing effects of ghrelin were attenuated by local administration of the GHS-R1A antagonist BIM28163 into the VTA , at a doP < 0.01) was not affected by VTA administration of the orexin A receptor antagonist SB334867 (P > 0.05) in mice  = 8.44, P = 0.005: n = 6–8) (P < 0.01) was not suppressed by i.p. injection of the opioid receptor antagonist naltrexone (P > 0.05) in mice  = 6.01, P = 0.003: n = 8) (The ghrelin-induced locomotor stimulation (n = 6–8) . Likewis: n = 8) .per se (P < 0.01) was attenuated by VTA administration of AP5 (P < 0.001) in mice  = 8.06, P < 0.001: n = 7-8). Moreover, systemic ghrelin increased dopamine release (P < 0.001) and pre-treatment with AP5 attenuated the ghrelin-induced increase in dopamine release compared to vehicle pre-treatment (P < 0.001)  = 19.98, P < 0.001; time F = 3.46, P < 0.001; treatment-time interaction F = 5.49, P < 0.001). This difference was evident at the time intervals 20–180 minutes (P < 0.001: n = 8–11).Intra-VTA administration of the NMDA receptor antagonist, AP5, abolished the ghrelin-induced locomotor stimulation and accumbal dopamine release , at a doper se . Specifiper se had any effect on locomotor activity . After the experiment the location of the probe and/or guide cannulae was verified and only mice with probe placement in the N.Acc. shell and/or cannula/e in the VTA were included in the statistical analysis . NeitherThe present study demonstrates that the stimulatory effects of peripheral ghrelin on locomotor stimulation and accumbal dopamine release involve VTA GHS-R1A signalling as both effects were suppressed by VTA administration of a GHS-R1A antagonist (BIM28163). Moreover, ghrelin's ability to activate the mesolimbic dopamine system was suppressed by pharmacological blockade of glutamatergic receptors but not by blockade of opioid or orexin A receptors. Thus, the locomotor stimulating effect of ghrelin was not affected by intra-VTA administration of the orexin A antagonist (SB334867) or by peripheral administration of an opioid receptor antagonist . Finally, ghrelin-induced locomotor stimulation as well as accumbal dopamine release were suppressed by VTA administration of an NMDA receptor antagonist (AP5). Taken together these data suggest that systemic ghrelin activates the mesolimbic dopamine system via GHS-R1A in the VTA and that glutamate rather than orexin or opioid signalling is required for ghrelin to stimulate the mesolimbic dopamine system.Given that transport of ghrelin across the blood-brain barrier into the brain is somewhat limited  and areaA receptors in the VTA does not mediate the increase in accumbal dopamine observed after food consumption induced by ghrelin  can be suppressed by an NMDA antagonist and are therefore likely to be under glutamatergic control. These data may have clinical implications since hyperghrelinemia is associated with addictive behaviours including compulsive overeating and alcohol use disorder (


Mutations within the gene encoding for the chloride ion channel CFTR results in cystic fibrosis, the most common autosomal recessive genetic disease in the Caucasian population. CFTR regulates absorption and secretion mechanisms across intestinal and airway mucosae. Although the intestinal phenotype can be clinically handled, chronic infection and inflammation of the lungs of CF patients remains the principal cause of morbidity and mortality. The aim of this Research Topic is to provide to the readers the most recent information available on “Strategies to circumvent the CFTR defect in cystic fibrosis.” The Research Topic is divided in three mains parts: the first part describes the CFTR structure, processing and regulation of the normal and mutant ion channels. Anna Patrick and Philip Thomas  review the molecular interactions leading to the complex folding process of the CFTR protein and how essential steps are disrupted in CFTR mutants. Soo Jung Kim and William Skach  detail the timing and coordination of specific folding steps of the native CFTR in and across the ER membrane. Colleen Weiler and Mitchell Drumm  broaden our views with genetic studies that have identified variant genes implicated in the clinical manifestations of CF. The second part of the Special topics discusses the molecular targets to rescue CFTR processing. Rebecca Chanoux and Ronald Rubenstein describe the current knowledge of the network of cellular chaperones that facilitate the folding and trafficking of CFTR to the plasma membrane. Then, a series of papers by the groups of Teresinha Leal  and John Hanrahan  report interesting observations regarding the potential of several inhibitors in correcting the CF phenotype at the level of ion transport and inflammation. Luigi Maiuri and collaborators  propose to target the intracellular environment in order to reestablish functional autophagy in CF epithelial cells. The third chapter is dedicated to new molecules that have already been developed or in development and able to rescue CFTR channel function by targeting mutant CFTR at the mRNA  and/or at the protein  levels. Finally, the Special Topic is concluded by an in-depth review by Christine Bear and collaborators . Importantly, the gaps in our knowledge regarding the mechanism of action of existing correctors, the unmet need to discover compounds which restore proper CFTR structure and function in CF affected tissues and new strategies for therapy development are discussed. We are convinced that we achieved a very interesting Special Topics thanks to the outstanding contributions of all authors. We are especially grateful to the authors for having believed in this project and accepted to share their knowledge. All the manuscripts have been peer-reviewed and we would like to thank the experts for helping us to reach an issue of high standard.


Prostate adenocarcinoma (CaP) is the most frequent cancer and second leading cause of cancer death among men in the Western world . Hh signOur work has shown that Hh signaling can be regulated in part by a protein normally expressed in the prostate, protease nexin 1  . PN1 is in vivo. Indeed, inducing expression of PN1 in several different prostate cancer cells was as effective in the inhibition of proliferation as treatment with the Hh-pathway inhibitor cyclopamine.Our new study features several components. Firstly, we show that PN1 inhibits Hh signaling by reduction in the hedgehog ligand, Sonic (SHH). This regulation is dependent upon uptake of PN1 through the LRP receptor and on the inhibitory activity of PN1. The immediate functional impact of PN1 is to reduce proliferation of prostate metastatic cells both in culture and Hh signaling inhibition also substantially reduced neo-angiogenesis. Xenografts from PC3 prostate cancer cells expressed high levels of SHH, producing small, disorganized vessels typical of tumour vasculature. These results are in line with other reports linking the Hedgehog pathway with the formation of new blood vessels and increases in tumor growth and metastasis . By contThe capacity to reduce tumor growth and vessel formation suggests a new physiological significance for PN1 and perhaps other serpins with similar functions. Interestingly, PN1 features some similarities to maspin, a tumor suppressor and angiogenesis blocker. Maspin has also been reported to reduce uPA-mediated CaP . HoweverAs a regulatory toggle, we have shown that PN1 is also a target of MMP9-mediated degradation. The localisation of PN1 in the ECM allows for its regulation by MMP9, an important factor in metastasis that can cleave collagen and basement membrane to drive this process. We extended these findings to show that higher MMP9 levels are inverse to PN1 levels and that this ratio is important in controlling hedgehog signaling. We demonstrate competing regulation of hedgehog signaling by PN1 and MMP9 in tissue culture, normal prostate tissues, xenografts, and orthotopic prostate cancers.The presence of PN1 in normal and BPH (benign prostate hypertrophy) tissues and decline in higher Gleason score samples may indicate that its loss is a marker of malignant transition in the prostate epithelium, amplifying Hh signaling. Indeed, one aim was to utilize Hh signalling as a stratifier of intermediate grade prostate cancer as the transitional status of these patients often makes treatment uncertain. Implications are that pharmacological inhibition of hedgehog signaling might be useful in this subset of patients. Thus, PN1 itself might be a potential therapeutic agent and, in combination with other SHH inhibitors, may be useful in the encouragement of CaP growth lag or inhibition. This work further indicates the potential important of hedgehog signaling in prostate cancer (Figure


Since the invention of scanning tunnelling microscopy (STM)  and atomAFM, especially, has turned out to be a versatile instrument, which can be operated in various environments, such as liquids, gases or vacuum. High and low temperature versions are available, which allow scientists to explore a large variety of materials. The strategy is not to prepare samples according to the requirements of the microscopy technique, but to perform experiments in its most native state, e.g., to study biological material in liquids. Questions to be addressed originate from almost all scientific areas. One example is the field of molecular electronics, where single molecules are investigated in order to perform specific tasks, e.g., molecular switches, molecular transistors or even molecular processors. In this area, STM and AFM have become essential tools to characterize the structure and function of molecules on surfaces.AFM has evolved considerably in the last few years, where new operation modes, such as non-contact force microscopy (nc-AFM), Kelvin probe force microscopy (KPFM) or friction force microscopy (FFM), were developed. One main focus is the high resolution capabilities of nc-AFM, which were drastically improved. Atomic resolution on metals, semiconductors  and insuAnother important development is related to nanomechanics, where phenomena, such as friction, wear, elasticity and plasticity are studied on an atomic scale. Atomic friction has been studied in great detail, where the main mechanism is related to atomic instabilities, which lead to the characteristic stick slip behaviour. The loading and velocitiy dependence were interpreted in terms of a thermally activated Prandtl–Tomlinson-model –6. The tAn important aspect of SPM is the possibility to modify surfaces. The probing tip can be either used to push or pull atoms, molecules or particles across surfaces. These experiments give information about the local bonding and to explore friction and wear mechanisms. Two different regimes were observed, which were related to the commensurability of the contacts . The manA schematic drawing of the family of scanning probe microscopes in Ernst MeyerBasel, December 2010


Bovine tuberculosis in domestic livestock such as cattle is an economically important disease with zoonotic potential, particularly in countries with emerging economies. We discuss the findings of recent epidemiological and immunological studies conducted in Ethiopia on host susceptibility differences between native zebu and the exotic Holstein–Friesian cattle that are increasingly part of the Ethiopian National herd, due to the drive to increase milk yields. These findings support the hypothesis that native Zebu cattle are more resistant to bovine tuberculosis. We also summarise the results of experimental infections that support the epidemiological data, and of laboratory experiments that suggest a role for the innate immune response, and in particular interleukin-6, in the outcome of bovine tuberculosis infection. These phylogenetically closely related bacteria share more than 99.9% chromosomal identity and they cause tuberculosis with similar pathology in various mammalian hosts Bovine tuberculosis (bTB) is a chronic bacterial disease of animals and humans. M. bovis are likely to also be an important risk factor for bTB in humans In animals, bTB can affect a wide range of species including both domestic and wildlife species Prevalence data on bTB in cattle are generally scarce, but official data reported by member countries of the OIE  suggest that the disease in domestic animals is widely distributed around the world and present on nearly all continents e.g. . Most inIn sub-Saharan Africa, animal production is facing new challenges since demographic growth, urbanization, and economic development are all contributing to the increasing demand for milk, meat, eggs and other animal products Ethiopia, at the Horn of Africa, is home to the largest livestock population in Africa with over 50 million cattle, of which more than 99% are the indigenous Zebu breed; the remaining animals are of imported exotic or mixed breeds A significant share of the work presented in this review is the fruit of an extensive project that was funded by the Wellcome Trust (UK) under their “Animal health in the developing world” initiative. Most of the studies were undertaken in Ethiopia to investigate (1) the prevalence and epidemiology of bTB in domestic and wildlife animals as well as risk factors for zoonotic transmission, (2) genetic aspects of host susceptibility to bTB, (3) efficacy of BCG vaccination in cattle, and (4) impact of bTB on the Ethiopian economy. The outcome of this research, and several stakeholder meetings held in Addis Ababa on bTB, has clearly had a significant impact on the perception and interest in the subject amongst policymakers, researchers and concerned farmers.2Bos taurus (taurine) cattle in West Africa (e.g. N’Dama), in contrast to trypano-susceptible cattle breeds like Boran , highlighted the fact that genetic traits in cattle can influence resistance to infectious diseases (reviewed in Theileria annulata) provides another example, with Holsteins (B. taurus) being extremely susceptible and several B. indicus breeds (e.g. Kenana cattle from Southern Sudan) showing a degree of resistance The natural trypano-tolerance of certain African B. indicus dairy cows were more resistant to experimental bTB than European, B. taurus, breeds. Although a more recent slaughterhouse study in India supported these findings that Indian breeds were less affected by bTB than pure European breeds like Jersey, Holstein, Friesians, Brown-Swiss Studies over the last 100 years have also indicated that differences exist in the relative susceptibility of different breeds of cattle to bTB. For example, Liston and Soparkar 3M. bovis challenge experiment using both Ankole and Zebu calves. His data confirmed his earlier epidemiological observations The incidence of bTB in East Africa, in particular in Uganda, in relation to cattle breeds, was studied by Carmichael in the 1930s B. indicus breeds had lower skin test prevalence (5.6%) compared to 86.4% in mainly Holsteins exotic breeds, with 13.9% prevalence in crosses . These studies did not compensate for the effect of animal husbandry conditions, although a recent study in Ethiopia In Ethiopia, PPD skin-testing of indicene and taurine cattle further supported the hypothesis that differences exist in their relative susceptibilities to bTB. For example, data collected in Eastern Shoa  indicates that diverse local 4To compensate for artefacts based on comparing cattle populations housed under different husbandry systems, our studies were focused on areas of Ethiopia where native zebu cattle (mainly the Arsi breed), Holstein cattle, as well as cross-breeds are kept under the same, low intensity management system. The study area chosen was North-western Shoa  where animals are kept together on pasture under low intensity farming conditions. The study area contained around 6000 smallholders with average herd sizes of about 10 animals . Half ofM. bovis-specific antigens ESAT-6 and CFP-10 M. bovis, the organism commonly associated with bTB, M. tuberculosis as well as non-tuberculosis-complex strains such as Mycobacterium avium avium were also found in tissue lesions In the same study, we were able to assess the pathology resulting from mycobacterial infection of cattle with respect to the extent and severity of visible pathology at post-mortem. This was assessed using a quantitative pathology scoring system developed for bTB vaccine research 5M. bovis infection. As it was logistically difficult to source Arsi zebus and conduct the experiment in Ethiopia, this experiment was conducted in South Africa using Holstein and Boran zebu cattle. Borans were selected because they are genetically closely related to Arsi zebus. Groups of six Boran and six Holstein–Friesian calves were each infected with a low dose of M. bovis and kept for 16 weeks, after which they were euthanized and their disease status established by post-mortem examination. The very low infective dose used To confirm experimentally the epidemiological data described above, a pilot experiment was conducted to test the response of taurine compared to indicene cattle to 6M. bovis/M. tuberculosis HSP65, CFP-10, MPB83, Ag85A and Rv3979c the most frequently recognised antigens . Serum antibody responses were directed towards MPB83 and MPB70, which are generally considered to be the sero-dominant antigens recognised after bTB infection of cattle + monocytes isolated from uninfected Holstein or Sahiwal cattle. These studies were undertaken in the UK using blood from Sahiwal zebus provided by Professor Glass . Monocytes were infected with either live M. bovis or M. tuberculosis at an MOI = 2, and a range of bovine cytokines and chemokines were measured in the culture supernatants. Nitric oxide production post-infection was also determined. We found no differences in the post-infection production of IL-1ß, IL-10 and IL-12 between Holstein or Sahiwal monocytes  8M. bovis infections are unclear, the fact that M. tuberculosis could be isolated from a sizable proportion of tuberculous cattle in our study area, could indicate a potential cattle-to-human transmission risk for M. tuberculosis.Our extensive field studies have provided epidemiological evidence that at least some zebu breeds are less susceptible to bTB and, if infected, present with less pathology than exotic breeds such as Holstein–Friesians. These results were supported by a small experimental infection study and by pilot studies on innate and acquired immune mechanisms that start to address the mechanisms underlying the observed differences in breed susceptibility to bTB. Apart from the obvious scientific aspects, the importance of these findings are of practical relevance, as they highlight the potential risks from increasing milk yield through the introduction of exotic cattle breeds that potentially are more susceptible to bTB then native zebus animals. Although the extent and risk of human zoonotic


Escherichia coli and Klebsiella pneumoniae bacteria isolated from clinical samples of patients attending Imam Reza and Ghaem University Hospitals, Mashhad, Northeast of Iran.Production of extended-spectrum beta-lactamases (ESBLs) by enteric bacteria continues to be a major problem in hospitals and community. ESBLs producing bacteria cause many serious infections including urinary tract infections, peritonitis, cholangitis and intra-abdominal abscess. The aim of this study was to determine the prevalence of ESBLs producing E. coli and 78 strains of K. pneumoniae were isolated from out-patients and hospitalized patients and they were examined by Oxoid combination disk test and PCR methods.During 2009 and 2010, 82 strains of E. coli and 56.1% of K. pneumoniae produced ESBLs. The frequency of SHV and TEM among the ESBLs producing isolates were 14.4% and 20.6%, respectively. Ratios of ESBLs positive isolates from out-patients to hospitalized patients were 24/33.We found that 43.9% of E. coli and K. pneumoniae is high in both study groups . Therefore it seems that continuous surveillance is essential to monitor the ESBLs producing microorganisms in hospitals and community. This study shows that the prevalence of ESBLs producing  Beta-lactam antibiotics are the most common drugs of any application in the treatment of bacterial infections . The betKlebsiella pneumoniae is the most common bacterial production of ESBLs; similarly, Escherichia coli is the other very important microorganism. ESBLs productions are rarely observed among other bacteria 8 cfu/ml (0.5 McFarland standard) were prepared in nutrient broth. Oxoid combination disk method was used for detection of ESBLs producing organisms. In this method the bacteria were cultured on a Muller-Hinton agar plate, then cefotaxim (30 µg), cefotaxim/clavulanate (10 µg), ceftazidime (30 µg) and ceftazidime/clavulanate (10 µg) disks  were placed on media in 20-30 mm with other disks. The plates were incubated for 18-24 hrs at 37 °C. ESBLs producing organisms were detected by an at least 5 mm increasing of zone around cefotaxim/clavulanate and ceftazidime/clavulanate at least 5 mm.For ESBL assay, bacterial suspensions with concentration of 1.5×10DNA extraction, PCR and sequencingThe colonies of ESBLs producing organisms were suspended in TE buffer and their DNA were extracted by simple boiling . 2, 0.2 mM each dNTP, and 5 u Taq DNA polymerase  in a total number of 50 μl of PCR reaction. Amplification of TEM and SHV genes was performed by following program: initial denaturation at 94 °C for 2 min and 35 cycles of 1 min at 94 °C, 30 sec at 52 °C and 1 min at 72 °C. Five min at 72 °C was considered for the final extension. Then, PCR products were analyzed by agarose gels electrophoresis. Eight PCR products from different types of the samples were sent for sequencing to Microgene Company, .The PCR method for detection of SHV and TEM genes was performed as described previously with minor modifications (14-15); briefly, specific primers for the genes  were used for PCR amplification that produced 471 bp and 861 bp PCR products for SHV and TEM genes, respectively. The PCR mixture consisted of 10 pmol of each primers, 1 μl DNA sample (3 μg/μl), 1.5 mM MgClE. coli n=82, K. pneumoniae n= 78) were collected; of these 57 isolates (35.6%) were ESBLs producing organisms (E. coli and 32 (20%) of isolated K. pneumoniae were ESBLs producing organisms.Among all the samples, urine samples were the most positive ones . In this study, a total number of 160 bacteria isolates .The distribution of ESBLs producing organisms based on out-patients and hospitalized patients are shown in In PCR method the distribution of TEM and SHV genes in isolated ESBLs producing organisms were 33 (20.6%) and 24 (14.4%), respectively , Table 3 sequenced and reported as Iranian strains in Gen Bank . The results obtained from sequencing was compared with the published sequences in Gen Bank and it showed that there was 99-100% (for SHV beta-lactamase gene) and 100% (for TEM-1b beta-lactamase gene) homology between our PCR products and those mentioned in Gen Bank.Eight PCR products from different kinds of samples wereE. coli and K. pneumoniae and to detect the presence of TEM and SHV genes among ESBL producing isolates. Based on the results of this study, the prevalence of ESBL producing E. coli and K. pneumoniae was high  and a total number of about 68.5% of ESBL producing isolated bacteria were TEM and/or SHV positive. A number of previous studies have showed the high prevalence of ESBLs producing E. coli and K. pneumoniae. In , the prevalence of ESBLs producing E. coli and K. pneumoniae isolates varies in different countries. In , the prevalence of the organisms was reported 46.51% in E. coli and 44.44% in K. pneumoniae isolates . In our study, the prevalence of ESBLs producing E. coli and K. pneumoniae was 15.6% and 20%, respectively.Reported ESBLs producing rates in et al in 2007 in Mashhad, the prevalence of ESBLs producing E. coli and K. pneumoniae was reported 57.5% and 61% and generally the prevalence of ESBLs producing organisms were 59.2% by Kirby-Baure disk diffusion method and the phenotypic disk confirmatory test. They also showed that the ratios of ESBLs producing isolates from hospitalized patients to out-patients were 94 to 28   . In Isfabacteria . Their ret al and Ramazanzadeh's results , and SHV (14.4%) genes by PCR. The TEM gene has high frequency compared to SHV gene; a fact which is similar to previous studies , 29 but  results , 31. AlsE. coli had 85.6% TEM, 69.2% SHV and 53.8% in both genes that was much higher than in our results (E. coli and K. pneumoniae) were also positive for SHV (52.7%) and TEM (32.4%) genes  genes ; and theIn short, the prevalence of ESBLs producing organisms in Mashhad is high. It seems necessary for clinicians and health care systems to be fully aware of ESBLs producing microorganisms. Also, the ESBLs production monitoring is recommended to avoid treatment failure and suitable infection control in Iran.


The understanding of the molecular mechanisms that underlie all stages of tumor progression in breast cancer (BC) represents an important goal in the biomedical research of this disease, particularly for the identification of more specific targeted therapies. In this context, BC preinvasive and precursor lesions represent a major dilemma. These lesions are well characterized under the phenotypic and genotypic profile, but it is not clear if they represent obligatory passages of a multistep process determining breast cancer evolution. In fact, the numerous cytogenetic and molecular alterations identified are not always representative of the progression into invasive phenotypes. BC precursor lesions include atypical ductal hyperplasia (ADH), atypical lobular hyperplasia (ALH), columnar cell lesions (CCLs), ductal carcinoma u (LCIS) .The molecular and morphological features of these lesions are indicative of a multistep model of breast tumorigenesis that provides a transition from normal epithelium to invasive carcinoma . SequentConsistent with the ‘clonal evolution’ hypothesis, there are numerous studies that show common genetic alterations among the precursors of low-grade BC (low-grade DCIS and ADH) .However, it has also been shown that these lesions do not have the same probability for progression into invasive carcinoma .Recently, as a result of the use of specific and sophisticated technologies, a wide range of molecular alterations associated with preneoplastic forms have been identified. Although they represent a ‘risk of evolution’, their real diagnostic and prognostic value remains unclear.Genotypic-phenotypic potential correlations are consistent with the hypothesis of a multistep evolution of precursor lesions. Several allelic imbalances have been identified in CCLs, located at 3p, 9p, 10q, 11q, 16q, 17p and 17q . MoreoveIn addition, ADH and DCIS exhibit extremely similar cytogenetic patterns consistent with the hypothesis of progression. These cytogenetic abnormalities largely involve the loss of 16q and 17p and the gain of 1q . By contWith regard to HER-2/c-erb2, hyperexpression/amplification has been detected in ADH and in 25% of LCIS cases if associated with invasive components . In addiE-cadherin is a molecular marker that is well characterized in BC precursor lesions, mainly as it is located in the 16q chromosomal region where losses are commonly described (Finally, epigenetic modifications have been reported to be associated with BC precursor lesions, in particular, gene methylation. E-cadherin hypermethylation has been highlighted in DCIS and the methylation pattern of this gene is consistent with the hypothesis that DCIS represents an IDC precursor .Molecular techniques, including DNA microarrays, immunohistochemistry and cytogenetic analysis, have made a significant contribution, in a number of cases, to the definition of specific molecular alterations that are common to several lesions at a ‘high risk’ of evolution .However, cellular models suitable for functional studies on the molecular alterations identified, are extremely limited and not representative of the numerous preinvasive/precursors lesions of BC. Furthermore, in the majority of cases, the variable molecular signatures of precancerous lesions are not associated with transformation to the invasive phenotype . TherefoThus, the existence of a multistep process of evolution of BC precursors lesions remains unknown, preventing the assignment of valid prognostic value to the molecular alterations associated.


The history of eponyms in obstetrics and gynecology is rich and long . EponymsDistinguishing whether the entry of Sonic the Hedgehog, the video game, cartoon, comic strip, and film character along with his great nemesis, Dr Ivo Robotnik, into the medical world is frivolous or dignified is largely a matter of personal taste, as well as is the use of other eponyms in medicine -6. HowevPresented is an admittedly cherry-picked list of fascinating food-related medical terms in obstetrics and gynecology to whet the appetite . We are


Dopamine transmission in the prefrontal cortex plays an important role in reward based learning, working memory and attention. Dopamine is thought to be released non-synaptically into the extracellular space and to reach distant receptors through diffusion. This simulation study examines how the dopamine signal might be decoded by the recipient neuron. The simulation was based on parameters from the literature and on our own quantified, structural data from macaque prefrontal area 10. The change in extracellular dopamine concentration was estimated at different distances from release sites and related to the affinity of the dopamine receptors. Due to the sparse and random distribution of release sites, a transient heterogeneous pattern of dopamine concentration emerges. Our simulation predicts, however, that at any point in the simulation volume there is sufficient dopamine to bind and activate high-affinity dopamine receptors. We propose that dopamine is broadcast to its distant receptors and any change from the local baseline concentration might be decoded by a transient change in the binding probability of dopamine receptors. Dopamine could thus provide a graduated ‘teaching’ signal to reinforce concurrently active synapses and cell assemblies. In conditions of highly reduced or highly elevated dopamine levels the simulations predict that relative changes in the dopamine signal can no longer be decoded, which might explain why cognitive deficits are observed in patients with Parkinson’s disease, or induced through drugs blocking dopamine reuptake. The dopamine signal in the prefrontal cortex (PFC) is crucial for working memory as well as for reinforcement learning To simulate the dopamine transmission in PFC, we used the parameters taken from published data and our own measurements of immunohistochemically labelled dopaminergic axons of layer 3 in prefrontal area 10 −1, the average dopamine concentration reached at steady state was 26 nM . The steady state level lies in the measured range of dopamine concentration in monkey prefrontal cortex At a tonic background firing of 5.6 Hz and a re-uptake constant of 1.5 sTo simulate the change in activity after an unexpected or omitted reward, we examined three different simulation conditions: an increase in firing up to 15 Hz for 150 ms followed by a pause of 0 Hz for 150 ms, an increased firing up to 26 Hz for 150 ms and a depressed activity of 0 Hz for 150 ms. As the number of dopaminergic neurons projecting to PFC with synchronous, phasic firing is not known, we varied the proportion of release sites responding to phasic activation from 10%, 25% and 50%. All the other release sites maintained tonic release. The measured values of all simulation conditions are summarized in The change in dopamine concentration over time was measured at three different measurement points: 1, 2 and 5 µm  away from every release site with phasic activity. The mean value (solid lines) with its standard deviation (dashed lines) is plotted along time for the respective simulation protocols in 3). The relative change in dopamine level varies strongly across the volume. For example, the dopamine concentration increases 14.4% on average over the volume after the phasic activity of 15 Hz . It has been reported that the dopamine level and innervation density in prefrontal areas of MPTP treated monkeys is reduced by 40–74% The dopamine re-uptake transporter is blocked by drugs such as Amphetamine, Methylphenidate  or Benzoyl-methyl-ecgonine (Cocaine) By simulating dopamine diffusion in prefrontal area 10 we could show that non-synaptic release of dopamine leads to local heterogeneity of dopamine concentration due to the low density of the release sites. During the phasic release of dopamine the ‘hot-spots’ enlarge and the overall dopamine level increases. Our results address important questions about how dopamine neurons encode their signal, how this signal is decoded by the recipient neurons in the PFC and also, by what means cell assemblies and individual synapses are modulated?In contrast to the striatum, where any given structure of the neuropil lies within a radius of 1 µm Although there is enough dopamine to activate receptors in their high-affinity state throughout the neuropil, the sparse innervation nevertheless results in dopamine concentrations that range over tens to several hundreds of nanomolar. ‘Hot-spots’ of increased dopamine concentration occur, but the simulations demonstrate that, unlike the dopamine level in the simulation of the striatum From the predictions of our simulation, we propose it is not the absolute value of dopamine, but the relative change of dopamine level that is detected against the baseline level of dopamine maintained by the tonic firing. Given that during tonic firing dopamine reaches a sufficient concentration to be bound by receptors in their high-affinity state, an increase or decrease in the concentration of dopamine principally affects the binding probability of the dopamine receptors and eventually the number of activated dopamine receptors. The crucial signal for the recipient site is hence not the absolute level, but whether there is more or less dopamine available relative to the local baseline level. Physiological recording of dopaminergic neurons indicate that they have three firing patterns: tonic firing as background activity, bursts of firing after the occurrence of an unexpected reward, and depressed activity of the neurons after an omission of an expected reward The two-state model of G-protein coupled receptors proposes that there is a dynamic balance between the high and low affinity state Our simulation of the Parkinsonian-like condition predicts that when only 30% of release sites are active, a detectable and effective dopamine concentration is provided only for receptors close to the release sites and not for the entire volume. On the other hand, when drugs block re-uptake the resultant increase in dopamine level to micromolar concentrations means that dopamine receptors can be occupied in their low affinity state. The dopamine receptors would then be bound to their ligand in both affinity states and no transient change in dopamine concentration would be detected as a change in binding probability of the dopamine receptors. For both very low numbers of release sites, or blocked re-uptake, a graduated dopamine signal would be no longer possible. Our simulation thus explains the inverted U-shaped relationship of dose-dependency for dopamine efficacy A reciprocal interaction has been described between the NMDA and D1 receptors Cx3Dp to include detailed neuronal structures with synaptic connectivity into the simulation volume, which would allow an exploration of the dynamics as well as plasticity of cell assemblies in relation to a change in dopamine concentration associated with a ‘teaching’ signal. Our simulations could also be extended to refining theoretical models of dopamine signal on working memory representation. We propose a central role of the dopamine receptors in the transmission of dopamine: dopamine is provided to all synapses in the volume and act on all structures in the neuropil that express the receptors necessary to receive the signal. Active synapses in a given cell assembly could be enhanced by means of the bidirectional interaction of dopamine-activated D1 receptors and NMDA receptors. Our study reveals the importance of a more detailed understanding of the dopamine receptors, their affinity states, and their interaction with other receptors and neurotransmitter systems.In the current study we were primarily interested in examining the spatiotemporal dynamic of the dopamine signal in the prefrontal cortex where the dopaminergic innervation is very sparse. However, these investigations could be extended within the simulation framework of Cx3Dp, a simulation framework that provides a three dimensional space to study biological processes with physical rules (http://www.ini.uzh.ch/~amw/seco/cx3d/). It was originally developed to model cortical growth and development Cx3Dp and is applied to different biological growth processes. For the simulation of dopamine diffusion in cortex it was necessary only to adjust the diffusion process with specific parameters for uptake and release of dopamine.The simulation of the diffusion was performed using 3 (64 µm on each side). In Cx3Dp, the space is divided into discrete compartments (1 µm3). Diffusion has been implemented with Fick’s Law as 1st order Euler step function with finite element methods across the compartments. The detailed composition of the neuropil was approximated by assuming the extracellular fraction of the volume was 23% and had a tortuosity of 1.6 The diffusion of dopamine is simulated in a cubic space representing an idealized block of cortical area 10, layer 3 with a volume of 0.0026 mm2 was determined from ultramicrographs in layer 3. Considering an average bouton diameter of 0.9 µm, a 3-D density of 0.0002/µm3 was obtained. A volume of 0.0026 mm3 contained 52 randomly located release sites with an average nearest neighbour distance of 9 µm. As the arbor of a single dopaminergic axon projecting to the cortex has not been characterised yet and the average interbouton distance or bouton density on a single dopamine axon is not known, we assumed release occurred independently at all sites.The density of release sites was measured on immunohistochemically stained tissue of monkey area 10 Dopaminergic cells have a tonic background firing rate of 5.3±1.5 Hz −1−1. For the purpose of the simulation the dopamine receptors and transporters were uniformly distributed in the space and thus the uptake was modelled to occur uniformly through the volume. We were interested in the activation of dopamine receptors D1 and D2 in relation to the dopamine concentration during tonic background activity and after the transient phasic release. D1 and D2 of the striatum are thought to be in high or low affinity states Immunohistochemical studies of dopaminergic receptors and transporters indicated that they are expressed distant to putative release sites or synapses All numeric values of the applied parameters and the corresponding references are given in the table below .


This study investigated whether semantic integration in discourse context could be influenced by topic structure using event-related brain potentials. Participants read discourses in which the last sentence contained a critical word that was either congruent or incongruent with the topic established in the first sentence. The intervening sentences between the first and the last sentence of the discourse either maintained or shifted the original topic. Results showed that incongruent words in topic-maintained discourses elicited an N400 effect that was broadly distributed over the scalp while those in topic-shifted discourses elicited an N400 effect that was lateralized to the right hemisphere and localized over central and posterior areas. Moreover, a late positivity effect was only elicited by incongruent words in topic-shifted discourses, but not in topic-maintained discourses. This suggests an important role for discourse structure in semantic integration, such that compared with topic-maintained discourses, the complexity of discourse structure in topic-shifted condition reduces the initial stage of semantic integration and enhances the later stage in which a mental representation is updated. Recently, a heated topic on discourse comprehension has been how readers interpret upcoming information with discourse context. Although it seems clear that semantic integration is rapidly influenced by global discourse context, whether and how semantic integration interacts with global discourse structure is less clear-cut. In the current study, we used event-related potentials (ERPs) to examine whether and how semantic integration is affected by the processing of topic-shifted structure as compared to topic-maintained structure of a discourse.A substantial body of studies has examined semantic integration in discourse context using eye movement and neurophysiological measures. Conflict between global discourse context and local information may result in longer reading times quick/slow”, which were designed such that the critical words were always coherent in the local sentence context. However, the critical words were either semantically coherent or incoherent with respect to the wider discourse . Relative to coherent control words , these discourse-dependent semantic anomalies elicited a large N400 effect that began at about 200 to 250 msec after word onset. The second experiment showed that when the same sentences were heard in isolation, the N400 effect disappeared. This suggests that the presence of the N400 effect was due to the mismatch between local information and discourse context.In two ERP experiments, van Berkum et al Note that semantic anomalies may not always give rise to an N400 effect, but may produce a P600-effect instead. The P600  is a positive deflection starting at about 400 ms and reaches its maximum around 600 ms post stimulus onset. It is typically related to the process of syntactic revision and repair in sentence processing when the sentences are not syntactically well-formed  participated in the experiment for cash. All were native speakers of Chinese and were right-handed with normal or corrected-to-normal vision. Three of the participants were rejected from the final analysis due to EEG-artifacts, experimenter error or technical problems with recording. Thus, 26 participants remained for subsequent analysis.M = 231.73, SD = 596.42; congruent: M = 170.63, SD = 312.13; t(159) = 1.25, p>.1) based on information provided by Beijing Institute of Language We constructed 160 sets of discourses. The first sentence was the lead-in sentence that established a topic. The last sentence contained a critical word which was either congruent or incongruent with the topic established in the first sentence. The intervening sentences between the first and the last sentence varied with discourse type (topic-shifted or topic-maintained). In the topic-shifted condition, the intervening sentences contained an overt shift in topic that departed away from the topic established in the first sentence. A topic shift was defined as a change in actor t(16) = 0.49, p>.05).Two pretests were conducted to assess the cloze probability and plausibility of the materials. In the first pretest, 17 participants were asked to write down the first word that came to mind to complete the discourses. The experimental stimuli were truncated before the critical word and randomly intermixed with the 80 filler discourses. Two lists were created in such a way that participants could only see the topic-maintained or the topic-shifted condition of the same item and there were an equal number of discourses from the two conditions. Results showed that no participant completed the sentences with the critical words that were used to create the incongruent condition. The mean cloze probability for the congruent words was 44.1% (SD = 7.8%) for the topic-shifted condition and 42.9% (SD = 7.6%) for the topic-maintained condition. A Paired-Samples t-test revealed no significant difference between the two structures  = 116.29, p<.001, η2partial = .86; mean ± SD = 3.92±0.52; 1.77±0.57; 3.96±0.60; 1.84±0.55 for topic-shifted/congruent, topic-shifted/incongruent, topic-maintained/congruent, and topic-maintained/incongruent respectively), while no main effect of discourse structure or interaction was found (Fs<1.4).In the second pretest, 20 participants were asked to rate the congruence of the discourses ranging from 1 (very implausible) to 5 (very plausible). The participants were assigned one of the four counterbalanced lists. Repeated-measures ANOVAs revealed a main effect of congruence . The EEG was sampled at 500 Hz with an amplifier bandpass of 0.01–100 Hz. Vertical EOG was recorded by electrodes placed above and below the left eye. The horizontal EOG was recorded via electrodes placed at the outer canthus of each eye. EEG data were referenced online to the left mastoid and re-referenced offline to the algebraic average of both mastoids. All electrode impedances were kept below 5 kΩ.EEG recordings were processed offline and ERPs were computed using the Edit program in Scan 4.3 . EEG and EOG records were screened for eye movements, electrode drifting, and excessive EEG amplitude exceeding ±75 µV. Contaminated trials were discarded . The analysis epoch started 200 ms prior to the onset of the critical word, and lasted for 1000 ms after the onset of the critical words. Averages were aligned to a 200-ms baseline preceding the critical word.Average waveforms were computed across all trials per condition for the 26 participants. Visual inspection of Analyses of variance (ANOVAs) were conducted with topic structure (topic-shifted vs. topic-maintained) and congruence (congruent vs. incongruent) as independent variables. In order to cover distributional differences in both the anterior–posterior as well as in the left–right dimensions, two topographical factors were also included in the ANOVAs. The first topographical factor was ant–pos, which had six levels: frontal, frontal-central, central, central-parietal, parietal, and occipital . The secAs shown in Fs<1, ps>.1). This indicates that subjects were reading the sentences attentively and they paid attention equally well to the topic-shifted and the topic-maintained discourses.Performance accuracy was above 85% for all participants for all experimental conditions. Repeated-measures ANOVA revealed neither the main effect of topic structure or congruence, nor the interaction between the two factors  = 1.91, p>.1, η2partial = 0.07; central: F  = 8.35, p<.01, η2partial = 0.25; central-parietal: F  = 3.79, p = .06, η2partial = 0.13; parietal: F  = 24.46, p<.001, η2partial = 0.50; and occipital: F  = 34.64, p<.001, η2partial = 0.58]. More importantly, there was a marginally significant three-way interaction between topic structure, congruence, and laterality.The overall analysis of N400 amplitude showed a main effect of congruence  = 3.44, p<.05, η2partial = 0.12]. Further simple effect analysis for the two-way interaction revealed that an N400 effect was only present at the right hemisphere . For the topic-maintained conditions, ANOVA with congruence and laterality as two within-participant factors revealed only a significant main effect of congruence . No interaction of congruence with laterality was observed . These findings indicated that the N400 effect elicited by topic-maintained condition was broadly distributed over the scalp.As the aim of the present study was to examine the impact of topic structure on semantic integration, we broke down the three-way interaction by the variable topic structure and calculated separate follow-up analyses within the topic-shifted (congruent vs. incongruent) and the topic-maintained (congruent vs. incongruent) conditions testing the variable congruence in left, medial, and right hemisphere. For the topic-shifted conditions, ANOVA with congruence and laterality as two within-participant factors revealed a marginally significant main effect of congruence  and a significant congruence×ant-pos interaction . Further simple effect analysis showed that an N400 effect was obtained at central, central-parietal, parietal, and occipital sites . For the topic-maintained condition, however, an ANOVA with congruence and ant–pos as two within-participant factors revealed only a significant main effect of congruence , indicating that the N400 effect elicited by the topic-maintained condition was distributed over the six ant–pos levels.Although the overall analysis did not indicate significant anterior to posterior differences in the scalp distribution of the N400 effect elicited by the topic-shifted and topic-maintained conditions, inspection of In summary, incongruent words elicited an N400 effect both in the topic-shifted condition and the topic-maintained condition. While the N400 effect in the topic-shifted condition was only found at the right hemisphere and localized to the central and posterior areas, the N400 effect elicited in the topic-maintained condition was found over the left, medial and right hemisphere and broadly distributed from frontal to posterior areas. This demonstrates that the N400 effect is broader in distribution in the topic-maintained condition than in the topic-shifted condition.p<.001, η2partial = 0.40], frontal-central , central , and central-parietal areas . For the congruence×laterality interaction, further simple effect analysis revealed that a late positivity was observed over the left  and medial sites .The overall analysis for the late positivity revealed a significant main effect of congruence  = 22.91, p<.001, η2partial = 0.48; frontal-central: F  = 22.20, p<.001, η2partial = 0.47; central: F  = 1.52, p>.1, η2partial = 0.06; central-parietal: F  = 6.31, p<.05, η2partial = 0.20; parietal: F  = 0.59, p>.1, η2partial = 0.02; and occipital: F  = 1.45, p>.1, η2partial = 0.06]. For the topic-maintained conditions, ANOVA with congruence and ant-pos as two within-participant factors revealed neither a significant main effect of congruence nor a significant interaction between congruence and ant-pos .More importantly, the overall analysis also revealed marginally significant topic structure×congruence and topic structure×congruence×ant-pos interactions. As with the analysis of the N400 effect, we resolved the three-way interaction by computing separate follow-up analyses within the topic-shifted and the topic-maintained conditions to examine the impact of topic structure on semantic integration. For the topic-shifted condition, an ANOVA with congruence and ant–pos as two within-participant factors revealed a significant congruence×ant-pos interaction [F  = 6.83, In summary, a late positivity effect was elicited by incongruent words in the topic-shifted condition, but not by those in the topic-maintained condition.This study aimed to investigate whether and how semantic integration is influenced by discourse structure. To examine this, we placed discourse-dependent anomalies in two kinds of discourse structure: topic-shifted and topic-maintained. Two important findings emerge from this study. Firstly, incongruent words in discourse context elicited an N400 effect followed by a late positivity effect, indicating that upcoming information was immediately related to the wider discourse context. Secondly and more importantly, we found that the N400 effect and the late positivity effect were modulated by discourse structure. The N400 effect is broader in distribution in the topic-maintained condition than in the topic-shifted condition. Moreover, the late positivity effect was only observed for incongruent words in the topic-shifted condition, but not for those in the topic-maintained condition. This suggests that shifts in discourse context reduce the initial stage of semantic integration and enhance the later stage in which a mental representation is updated.The result that incongruent words in discourse context elicited a classical N400 effect relative to congruent words is quite compatible with the current literature about this component. It has been suggested that the N400 is sensitive to the ease with which a word can be related to the wider discourse In addition to the classical N400 effect, a frontalcentral late positivity effect was also elicited by incongruent words in the topic-shifted condition. The finding of a frontalcentral late positivity has been reported in the literature. Friederici, Hahne, & Saddy The primary aim of this study was to investigate whether topic structure influences semantic integration. The results obtained in the EPRs revealed that the incongruent words in the topic-maintained discourses elicited a broadly distributed N400 effect while those in topic-shifted discourses elicited a restrictedly distributed N400 effect followed by a late positivity effect. These results suggest that discourse structure modulates both the initial stage of semantic integration and the later stage of discourse updating.Previous studies have suggested that a topic-maintained structure aids in keeping related concepts active in working memory while a shift in topic creates an interference effect that reduces the accessibility of relevant information Compared with topic-maintained discourses, a late positivity effect was found in response to incongruent words in topic-shifted discourses, but not in topic-maintained discourse. This may be due to the fact that the construction of a coherent representation involves more work in topic-shifted discourses than in topic-maintained discourses


Bothrops jararaca and Lachesis muta. In vitro assays showed that these compounds were impaired in a concentration-dependent manner, the fibrinogen or plasma clotting, hemolysis, and proteolysis produced by both venoms. Moreover, these compounds inhibited biological effects in vivo  as well. Mice treated with these compounds were fully protected from hemorrhagic lesions caused by such venoms. But, only the B. jararaca edema-inducing activity was neutralized by the triazoles. So the inhibitory effect of triazoles derivatives against some in vitro and in vivo biological assays of snake venoms points to promising aspects that may indicate them as molecular models to improve the production of effective antivenom or to complement antivenom neutralization, especially the local pathological effects, which are partially neutralized by antivenoms.Snake venoms are complex mixtures of proteins of both enzymes and nonenzymes, which are responsible for producing several biological effects. Human envenomation by snake bites particularly those of the viperid family induces a complex pathophysiological picture characterized by spectacular changes in hemostasis and frequently hemorrhage is also seen. The present work reports the ability of six of a series of 1,2,3-triazole derivatives to inhibit some pharmacological effects caused by the venoms of  The venomous pit vipers Bothrops jararaca and Lachesis muta are responsible for accidents involving humans in several regions of South America. While B. jararaca is found in southern Brazil, Paraguay, and northern Argentina, L. muta is distributed in the equatorial forests east of the Andes, ranging from eastern Ecuador, Colombia, Peru, northern Bolivia and eastern and northern Venezuela, to Guyana, French Guyana, Surinam, and northern Brazil. Within their range, they are often abundant and are important cause of snakebites -1-(p-chlorophenyl)-1H--triazole-4-carbohydrazide against in vivo and in vitro activities of Bothrops jararaca and Lachesis muta venoms.The aim of this work was to evaluate the ability of these six 1,2,3-triazole derivatives based on Bothrops jararaca, Lachesis muta lyophilized venoms, and anti-Lachesis or anti-Bothropic antivenom were provided from Fundação Ezequiel Dias, Belo Horizonte, MG, Brazil, and stored at −20°C until assays. Dimethylsufoxide (DMSO), bovine fibrinogen, and azocasein were obtained from Sigma Chemical Co. All other reagents were of the best grade available.H--triazole-4-carboxylic acid ethyl esters derivatives were synthesized according to our previous report -triazole-4-carboxylic acid ethyl esters to neutralize some in vitro  and in vivo  activities caused by B. jararaca and L. muta venoms, since previous results indicated that these six derivatives inhibited hemolysis induced by L. muta venom, but with different potencies [B. jararaca (50 μg/mL) and L. muta (15 μg/mL) venom  venom . The inhB. jararaca and L. muta venom hydrolyzed azocasein in a concentration-dependent manner with an EC of 20 μg/mL and 6 μg/mL, respectively (data not shown). The derivatives inhibited proteolysis induced by B. jararaca or L. muta , the clotting of fibrinogen induced by the venoms of B. jararaca (40 μg/mL) or L. muta (10 μg/mL). It seemed that derivatives inhibited more efficiently L. muta-inducing clotting than B. jararaca. At the highest concentration (94 μM), the derivatives 1, 2, 3, 5 and 6 prevented L. muta clotting  were put all together and incubated with either B. jararaca or L. muta venom, the clotting time was delayed two-fold. It was noticed that, if derivative 2 or 6 was removed from mixture, inhibitory effect upon clotting was not observed. Moreover, the derivatives also prevented clotting induced by venoms when plasma was used. Neither DMSO  nor saline solution interfered with coagulation processes.As seen in clotting , while taca once . At concB. jararaca (12 mg/Kg) or L. muta (20 mg/Kg) venom produced a hemorrhage halo of 20 mm in mice. Such a halo represents two MHD of venoms. When each venom was mixed with derivatives (90 μM) and then injected into mice, a complete protection from hemorrhage was seen (data not shown). In contrast, previous results showed that antilaquetic serum did not inhibit hemorrhage induced by L. muta venom [B. jararaca (L. muta (μM). The triazole derivatives 1, 2, and 4 inhibited above 80% the edema induced by B. jararaca, whereas derivatives 3, 5, and 6 inhibited around 70%  pointing for good theoretical biodisponibility [In conclusion, 1-arylsulfonylamino-5-methyl-1erformed . The ders than 5 . The resnibility .