A Brazilian glycoprotein E-negative bovine herpesvirus type 1.2a (BHV-1.2a) mutant is attenuated for cattle and induces protection against wild-type virus challenge

The authors previously reported the construction of a glycoprotein E-deleted (gE-) mutant of bovine herpesvirus type 1.2a (BHV-1.2a). This mutant, 265gE-, was designed as a vaccinal strain for differential vaccines, allowing the distinction between vaccinated and naturally infected cattle. In order to determine the safety and efficacy of this candidate vaccine virus, a group of calves was inoculated with 265gE-. The virus was detected in secretions of inoculated calves to lower titres and for a shorter period than the parental virus inoculated in control calves. Twenty one days after inoculation, the calves were challenged with the wild type parental virus. Only mild signs of infection were detected on vaccinated calves, whereas non-vaccinated controls displayed intense rhinotracheitis and shed virus for longer and to higher titres than vaccinated calves. Six months after vaccination, both vaccinated and control groups were subjected to reactivation of potentially latent virus. The mutant 265gE- could not be reactivated from vaccinated calves. The clinical signs observed, following the reactivation of the parental virus, were again much milder on vaccinated than on non-vaccinated calves. Moreover, parental virus shedding was considerably reduced on vaccinated calves at reactivation. In view of its attenuation, immunogenicity and protective effect upon challenge and reactivation with a virulent BHV-1, the mutant 265gE- was shown to be suitable for use as a BHV-1 differential vaccine virus.

Experimental infection of rabbits with a recombinant bovine herpesvirus type 5 (BoHV-5) gI, gE and US9-negative

Bovine herpesvirus type 5 (BoHV-5) is a major cause of viral meningoencephalitis in cattle. The expression of different viral proteins has been associated with BoHV-5 neuropathogenesis. Among these, gI, gE and US9 have been considered essential for the production of neurological disease in infected animals. To evaluate the role of gI, gE and US9 in neurovirulence, a recombinant from which the respective genes were deleted (BoHV-5 gI-/gE-/US9-) was constructed and inoculated in rabbits of two age groups (four and eight weeks-old). When the recombinant virus was inoculated through the paranasal sinuses of four weeks-old rabbits, neurological disease was observed and death was the outcome in 4 out of 13 (30.7 %) animals, whereas clinical signs and death were observed in 11/13 (84.6%) of rabbits infected with the parental virus. In eight weeks-old rabbits, the BoHV-5 gI-/gE-/US9- did not induce clinically apparent disease and could not be reactivated after dexamethasone administration, whereas wild type BoHV-5 caused disease in 55.5% of the animals and was reactivated. These findings reveal that the simultaneous deletion of gI, gE and US9 genes did reduce but did not completely abolish the neurovirulence of BoHV-5 in rabbits, indicating that other viral genes may also play a role in the induction of neurological disease.

A thymidine kinase-negative bovine herpesvirus 5 is highly attenuated for rabbits, but is neuroinvasive and establishes latent infection

Mutant viral strains deleted in non-essential genes represent useful tools to study the function of specific gene products in the biology of the virus. We herein describe an investigation on the phenotype of a bovine herpesvirus 5 (BoHV-5) recombinant deleted in the gene encoding the enzyme thymidine kinase (TK) in rabbits, with special emphasis to neuroinvasiveness and the ability to establish and reactivate latent infection. Rabbits inoculated with the parental virus (SV-507/99) (n=18) at a low titer (10(5.5)TCID50) shed virus in nasal secretions in titers up to 10(4.5)TCID50 for up to 12 days (average: 9.8 days [5-12]) and 5/ 16 developed neurological disease and were euthanized in extremis. Rabbits inoculated with the recombinant BoHV-5TKΔ at a high dose (10(7.1)TCID50) also shed virus in nasal secretions, yet to lower titers (maximum: 10(2.3)TCID50) and for a shorter period (average: 6.6 days [2-11]) and remained healthy. PCR examination of brain sections of inoculated rabbits at day 6 post-infection (pi) revealed a widespread distribution of the parental virus, whereas DNA of the recombinant BoHV-5TKΔ-was detected only in the trigeminal ganglia [TG] and olfactory bulbs [OB]. Nevertheless, during latent infection (52pi), DNA of the recombinant virus was detected in the TGs, OBs and also in other areas of the brain, demonstrating the ability of the virus to invade the brain. Dexamethasone (Dx) administration at day 65 pi was followed by virus reactivation and shedding by 5/8 rabbits inoculated with the parental strain (mean duration of 4.2 days [1 - 9]) and by none of seven rabbits inoculated with the recombinant virus. Again, PCR examination at day 30 post-Dx treatment revealed the presence of latent DNA in the TGs, OBs and in other areas of the brain of both groups. Taken together, these results confirm that the recombinant BoHV-5TKΔ is highly attenuated for rabbits. It shows a reduced ability to replicate in the nose but retains the ability to invade the brain and to establish latent infection. Additional studies are underway to determine the biological and molecular mechanisms underlying the inability of BoHV-5TKΔ to reactivate from latency.

Metabolic evaluation of dairy cows submitted to three different strategies to decrease the effects of negative energy balance in early postpartum

In early lactation dairy cattle suffer metabolic alterations caused by negative energy balance, which predisposes to fatty liver and ketosis. The aim of this study was to evaluate the metabolic condition of high yielding dairy cows subjected to three treatments for preventing severe lipomobilization and ketosis in early lactation. Fifty four multiparous Holstein cows yielding >30 L/day were divided into four groups: control (CN= no treatment), glucose precursor (PG= propylene-glycol), hepatic protector (Mp= Mercepton®), and energy supplement with salts of linolenic and linoleic faty acids (Mg-E= Megalac-E®). Treatments were administrated randomly at moment of calving until 8 weeks postpartum. Blood samples were collected on days 1, 7, 14, 21, 28, 35, 42 and 49 postpartum. Body condition score (BCS) was evaluated at the same periods and milk yield was recorded at 2nd, 4th, 5th, 6th, 7th, and 8th weeks of lactation. Concentrations of non-esterified fatty acids (NEFA), albumin, AST, ß-hydroxybutyrate (BHBA), cholesterol, glucose, total protein, urea and triglycerides were analyzed in blood samples. Cut-off points for subclinical ketosis were defined when BHBA >1.4 mmol/L and NEFA >0.7 mmol/L. General occurrence of subclinical ketosis was 24% during the period. An ascendant curve of cholesterol and glucose was observed from the 1st to the 8th week of lactation, while any tendency was observed with BHBA and NEFA, although differences among treatments were detected (p<0.05). BCS decreased from a mean of 3.85 at 1st week to 2.53 at 8th week of lactation (p=0.001). Milk yield was higher in the Mg-E group compared with the other treatment groups (p<0.05) Compared with the CN group, the treatments with Mp and PG did not show significant differences in blood biochemistry and milk yield. Cows receiving PG and Mg-E showed higher values of BHBA and NEFA (P<0.05), indicating accentuated lipomobilization. Supplementation with Mg-E also resulted in significant higher concentrations of cholesterol, BHBA, urea, AST and lower values of glycemia. This performance may be explained by the highest milk yield observed with this treatment. Treatments with PG and Mp did not improve milk yield, compared with control cows, but did not show metabolic evidence of ketosis, fat mobilization or fatty liver. These results suggest that treatment with Mg-E improves milk production but induces a higher negative energy balance leading to moderated lipomobilization and ketone bodies production, increasing the risk of fatty liver.

Sivistyksestä sivilisaatioon. Kulttuurikäsitys J. V. Snellmanin historiallisessa ajattelussa

From Bildung to Civilisation. Conception of Culture in J. V. Snellman’s Historical Thinking The research explores Johan Vilhelm Snellman’s (1806–1881) conception of culture in the context of his historical thinking. Snellman was a Finnish, Swedish-speaking journalist, teacher and thinker, who held a central position in the Finnish national discourse during the nineteenth century. He has been considered as one of the leading theorists of a Finnish nation, writing widely about the themes such as the advancement of the national education, Finnish language and culture. Snellman is already a widely studied person in Finnish intellectual history, often characterised as a follower of G. W. F. Hegel’s philosophical system. My own research introduces a new kind of approach on Snellman’s texts, emphasising the conceptual level of his thought. With this approach, my aim is to broaden the Finnish research tradition on conceptual history. I consider my study as a cultural history of concepts, belonging also to the field of intellectual history. My focus is on one hand on the close reading of Snellman’s texts and on the other hand on contextualising his texts to the European intellectual tradition of the time. A key concept of Snellman’s theoretical thinking is his concept of bildning, which can be considered as a Swedish counterpart of the German concept of Bildung. The Swedish word incorporated all the main elements of the German concept. It could mean education or the so-called high culture, but most fundamentally it was about the self-formation of the individual. This is also the context in which Snellman’s concept of bildning has often been interpreted. In the study, I use the concept of bildning as a starting point of my research but I broaden my focus on the cognate concepts such as culture (kultur), spirit (anda) and civilisation. The purpose of my study is thus to illustrate how Snellman used and modified these concepts and from these observations to draw a conclusion about the nature of his conception of culture. Snellman was an early Finnish philosopher of history but also interested in the practice of the writing of history. He did not write any historical presentations himself but followed the publications in the field of history and introduced European historical writing to the Finnish, Swedish-speaking reading audience in his newspapers. The primary source material consists of different types of Snellman’s texts, including philosophical writings, lecture material, newspaper articles and private letters. I’m reading Snellman’s texts in the context of other texts produced both by his Finnish predecessors and contemporaries and by Swedish, German and French writers. Snellman’s principal philosophical works, Versuch einer spekulativen Entwicklung der Idee der Persönlichkeit (1841) and Läran om staten (1842), were both written abroad. Both of the works were contributions to contemporary debates on the international level, especially in Germany and Sweden. During the 1840s and 1850s Snellman had two newspapers of his own, Saima and Litteraturblad, which were directed towards the Swedish-speaking educated class. Both of the newspapers were very popular and their circulations were among the largest of their day in Finland. The topics of his articles and reviews covered literature, poetry, philosophy and education as well as issues concerning the economic, industrial and technical development in Finland. In his newspapers Snellman not only brought forth his own ideas but also spread the knowledge of European events and ideas to his readers. He followed very carefully the cultural and political situation in Western Europe. He also followed European magazines and newspapers and was well acquainted with German, French and also English literature – and of course Swedish literature to with which he had the closest ties. In his newspapers Snellman wrote countless number of literary reviews and critics, introducing his readers to European literature. The study consists of three main chapters in which I explore my research question in three different, yet overlapping contexts. In the first of these chapters, I analyse Snellman’n theoretical thinking and his concepts of bildning, kultur, anda and civilisation in the context of earlier cultural discourse in Finland as well as the tradition of German idealistic philosophy and neo-humanism. With the Finnish cultural discourse I refer to the early cultural discussion in Finland, which emerged after the year 1809, when Finland became an autonomous entity of its own as a Grand Duchy of Russia. Scholars of the Academy of Turku opened a discussion on the themes such as the state of national consciousness, the need for national education and the development of the Finnish language as a national language of Finland. Many of these academics were also Snellman’s teachers in the early years of his academic career and Snellman clearly formulated his own ideas in the footsteps of these Finnish predecessors. In his theoretical thinking Snellman was a collectivist; according to him an individual should always be understood in connection with the society, its values and manners, as well as to the traditions of a culture where an individual belongs to. In his philosophy of the human spirit Snellman was in many ways a Hegelian but his notion of education or ‘bildning’ includes also elements that connect him with the wider tradition of German intellectual history, namely the neo-humanist tradition and, at least to some extent, to the terminology of J. G. Herder or J. G. Fichte, for example. In this chapter, I also explore Snellman’s theory of history. In his historical thinking Snellman was an idealist, believing in the historical development of the human spirit (Geist in German language). One can characterise his theory of history by stating that it is a mixture of a Hegelian triumph of the spirit and Herderian emphasis on humanity (Humanität) and the relative nature of ‘Bildung’. For Snellman, the process of ‘bildning’ or ‘Bildung’ is being realised in historical development through the actions of human beings. Snellman believed in the historical development of the human civilization. Still Snellman himself considered that he had abandoned Hegel’s idea about the process of world history. Snellman – rightly or wrongly – criticised Hegel of emphasising the universal end of history (the realisation of the freedom of spirit) at the expense of the historical plurality and the freedom of each historical era. Snellman accused Hegel of neglecting the value and independency of different historical cultures and periods by imposing the abstract norm, the fulfilment of the freedom of spirit, as the ultimate goal of history. The historicist in Snellman believed in the individuality of each historical period; each historical era or culture had values, traditions and modes of thought of its own. This historicist in Snellman could not accept the talk about one measure or the end of history. On the other hand Snellman was also a universalist. He believed that mankind had a common task and that task was the development of ‘Bildung’, freedom or humanity. The second main chapter consists of two parts. In the first part, I explore the Finnish nationalistic discourse from the cultural point of view by analysing the notions such as a nation, national spirit or national language and showing how Snellman formulated his own ideas in a dialogic situation, participating in the Finnish discourse but also reacting to international discussions on the themes of the nation and nationality. For Snellman nationality was to a great extent the collective knowledge and customs or practices of the nation. Snellman stated that nationality is to be considered as a form of ‘bildning’. This could be seen not simply as affection for the fatherland but also for the mental identity of the nation, its ways of thinking, its practices, national language, customs and laws, the history of the nation. The simplest definition of nationality that Snellman gives is that nationality is the social life of the people. In the second part of the chapter I exam Snellman’s historical thinking and his understanding about historical development, interaction between different nations and cultures in the course of history, as well as the question of historical change; how do cultures or civilisations develop and who are the creators of culture? Snellman did not believe in one dominating culture but understood the course of history as a dialogue between different cultures. On the other hand, his views are very Eurocentric – here he follows the ideas of Hegel or for example the French historian François Guizot – for Snellman Europe represented the virtue of pluralism; in Europe one could see the diversity of cultures which, on the other hand, were fundamentally based on a common Christian tradition. In the third main chapter, my focus is on the writing of history, more precisely on Snellman’s ideas on the nature of history as a science and on the proper way of writing historical presentations. Snellman wrote critics on the works of history and introduced his readers to the writing of history especially in France, Sweden and German-speaking area – in some extend also in Britain. Snellman’s collectivistic view becomes evident also in his reviews on historical writing. For Snellman history was not about the actions of the states and their heads, nor about the records of ruling families and battles fought. He repeatedly stressed that history is a discipline that seeks to provide a total view of a phenomenon. A historian should not only collect information on historical events, since this information touches only the surface of a certain epoch or civilisation; he has to understand an epoch as totality. This required an understanding about the major contours in history, connections between civilisations and an awareness of significant turning points in historical development. In addition, it required a holistic understanding about a certain culture or historical era, including also the so-called inner life of a specific nation, a common people and their ways of life. Snellman wrote explicitly about ‘cultural history’ in his texts, referring to this kind of broad understanding of a society. In historical writing Snellman found this kind of broader view from the works of the French historians such as François Guizot and Jules Michelet. In all of these chapters, I elaborate the conceptual dimension of Snellman’s historical thinking. In my study I argue that Snellman not only adopted the German concepts of Bildung or Kultur in his own thinking but also developed the Swedish concepts in a way that include personal and innovative aspects. Snellman’s concept of bildning is not only a translation from ‘Bildung’ but he uses the Swedish concept in a versatile way that includes both the moral aspect of human development and social dimension of a human life. Along with ‘bildning’ Snellman used also the terms ‘kultur’ and ‘civilisation’ when referring to the totality of a certain nation or historical era, including both the so-called high culture (arts, science, religion) and the modes of thought as well as ways of life of the people as a whole. Unlike many of his Finnish contemporaries, Snellman did not use civilisation as a negative concept, lacking the moral essence of German term ‘Bildung’ or ‘Kultur’. Instead, for Snellman civilisation was a neutral term and here he comes close to the French tradition of using the term. In the study I argue that Snellman’s conception of culture in fact includes a synthesis of the German tradition of ‘Bildung’ and the French tradition of ‘civilisation’.

Antimicrobial resistance and detection of mecA and blaZ genes in coagulase-negative Staphylococcus isolated from bovine mastitis

The present study evaluated the pheno- and genotypical antimicrobial resistance profile of coagulase-negative Staphylococcus (CNS) species isolated from dairy cows milk, specially concerning to oxacillin. Of 100 CNS isolates, the S. xylosus was the prevalent species, followed by S. cohnii, S. hominis, S. capitis and S. haemolyticus. Only 6% were phenotypically susceptible to the antimicrobial agents tested in disk diffusion assay. Penicillin and ampicillin resistance rates were significantly higher than others antimicrobials. Four isolates were positive to mecA gene (4%), all represented by the S. xylosus species. The blaZ gene was detected in 16% of the isolates (16/100). It was noticed that all mecA + were also positive to this gene and the presence of both genes was correlated to phenotypic beta-lactamic resistance. We conclude that CNS species from bovine milk presented significantly distinct antimicrobial resistance profiles, evaluated by phenotypic and genotypic tests, which has implications for treatment and management decisions.

Beta-lactamase detection in Staphylococcus aureus and coagulase-negative Staphylococcus isolated from bovine mastitis

The objectives of the study were to evaluate the presence/production of beta-lactamases by both phenotypic and genotypic methods, verify whether results are dependent of bacteria type (Staphylococcus aureus versus coagulase-negative Staphylococcus - CNS) and verify the agreement between tests. A total of 200 bacteria samples from 21 different herds were enrolled, being 100 CNS and 100 S. aureus. Beta-lactamase presence/detection was performed by different tests (PCR, clover leaf test - CLT, Nitrocefin disk, and in vitro resistance to penicillin). Results of all tests were not dependent of bacteria type (CNS or S. aureus). Several S. aureus beta-lactamase producing isolates were from the same herd. Phenotypic tests excluding in vitro resistance to penicillin showed a strong association measured by the kappa coefficient for both bacteria species. Nitrocefin and CLT are more reliable tests for detecting beta-lactamase production in staphylococci.

Chaos in a Two-Degree of Freedom Duffing Oscillator

High dimensional dynamical systems has intricate behavior either on temporal or on spatial evolution properties. Nevertheless, most of the work on chaotic dynamics has been concentrated on temporal behavior of low-dimensional systems. This contribution is concerned with the chaotic response of a two-degree of freedom Duffing oscillator. Since the equations of motion are associated with a five-dimensional system, the analysis is performed by considering two Duffing oscillators, both with single-degree of freedom, coupled by a spring-dashpot system. With this assumption, it is possible to analyze the transmissibility of motion between the two oscillators.

„Und frei erklär' ich alle meine Knechte“ : Höflichkeit als Ausdruck sozialer Utopie in Schillers Drama Wilhelm Tell.

My dissertation is an interdisciplinary study of the relationship between the Swiss peasants and nobles in Friedrich Schiller’s play Wilhelm Tell (1804). Changes to this relationship are of particular interest. Communication in the play is examined via a micro-analysis based on Penelope Brown’s and Stephen C. Levinson’s theoretical framework Politeness: Some universals in language usage. (1978, 1987). Brown and Levinson distinguish between positivepoliteness and negative-politeness strategies and their distinction is useful for my argument, since they claim that the use of positive-politeness strategies reflects the speaker’s intention to minimize the distance between the interlocutors (Brown and Levinson 1987: 103). Negative-politeness strategies, by contrast, result in social distancing (Brown and Levinson 1987: 130). In accordance with Brown’s and Levinson’s theory, it can be argued that the distribution of positive-politeness and negative-politeness strategies reflects changes in the distance between the fictional interlocutors as representatives of their social classes in Wilhelm Tell. The analysis of the communication in the play highlights that existing conflicts within the social groups are resolved and replaced with solidarity (cf. ‘Claim ’common ground’ and ‘Claim in-group membership with H’) before the peasants and noblemen appear on stage simultaneously in III, 3. In the scene in question, Geßler forces Tell to shoot the apple off his son’s head. Although both nobles and peasants are present, they do not communicate with each other. Thus, communication between the social classes occurs for the first time in IV, 2. This scene is crucial with regard to the changes in the relationship between the social classes in the play. The younger generation, with Rudenz as a representative of the nobility and Melchthal as a representative of the peasants, break with the prevailing conventions and initiate a new type of cooperation based on mutual helping and equality in the right and the duty to protect the country from the violent oppressors representing Habsburg (cf. ‘Convey that S and H are cooperators’, ‘Claim reflexivity’ and ‘Claim reciprocity’). The linguistic analysis reveals the crucial role of the Swiss nobility in the development of the social utopia, as well as the non-contribution of Wilhelm Tell. In fact, Tell never communicates with the Swiss noblemen. The role of the nobility and the role of Wilhelm Tell are further investigated in a comparison between Schiller’s Wilhelm Tell and four of Schiller’s historical sources. These sources, which contain earlier, non-dramatic versions of the establishment of the Swiss Confederation, are the following: Kronica von der loblichen Eydtgnoschaft compiled by Petermann Etterlin (1507), Gemeiner loblicher Eydgnoschafft Stetten Landen vnd Völckeren Chronik wirdiger thaaten beschreybung (1548) written by Johann Stumpf, Chronicon Helveticum of Aegidius Tschudi (the publication of Iselin from 1734 and 1736) and Johannes von Müller’s Geschichten schweizerischer Eidgenossenschaft (1786). Thecomparison sheds light on the fact that both Tschudi and von Müller emphasize unity among the Swiss and cooperation in their attempt to defeat the enemy. However, Schiller has reinforced the role of the nobility in the cooperation. In addition, the comparison between Schiller’s play and the historical sources reveals profound differences with regard to the role of Wilhelm Tell. In all of Schiller’s sources, Tell is present at Rütli, whereas he is absent from Rütli in Schiller’s play. In the play in general, Tell is conspicuously separated from the other peasants. Explanations of my linguistic results, which correspond to the above mentioned differences between the roles of the figures in Wilhelm Tell and the depictions in Schiller’s sources, are found by comparing the path of Schiller’s Swiss towards an egalitarian perspective with the kind of social evolution depicted by the German philosopher Johann Benjamin Erhard in his essay Über das Recht des Volks zu einer Revolution (1795). Aiming at considering Schiller’s social utopia within the larger cultural framework of the German response to the French Revolution, the comparison suggests that both authors depict the need for social change in terms of a change in social hierarchies (Erhard 1970: 95-96, cf. Foi 2005: 225). Erhard’s essay thus helps explain the political intention of Schiller’s play to keep the ideals of the French Revolution as crucial aims but profoundly change the means towards freedom and equality. In his attempt to claim the political resistance of the nobles and peasants as just, however, Schiller sacrificed the figure of Wilhelm Tell. Guilty of the murder of Geßler, Tell was no longer suitable for the righteous revolution imagined by Schiller (cf. Bloch 2008: 215 and Schulz 2005: 228). This explains Tell’s absence in the Rütli scene, his isolation in the plot, as well as his non-contribution to the social utopia. Together, the linguistic analysis and contextualisation of Schiller’s play support my hypothesis that Wilhelm Tell describes a process of change in the relationship between peasants and nobles. The interdisciplinary approach to the topic proved to be fruitful for all areas of the research involved.

Negative refractive index materials and propagation

Negative refractive index materials and propagation of electromagnetic waves in them started to draw attention of scientists not so long ago. This review highlights historically important and recent papers on practical and theoretical aspects related to these issues. Namely, basic properties and peculiarities of such materials related to both their design and wave propagation in them, experimental verification of predictions theoretically made for them, possible practical applications and prospects in this area are considered.

Dual light barrier method for six degrees of freedom tool center point calibration of an industrial robot

Tool center point calibration is a known problem in industrial robotics. The major focus of academic research is to enhance the accuracy and repeatability of next generation robots. However, operators of currently available robots are working within the limits of the robot´s repeatability and require calibration methods suitable for these basic applications. This study was conducted in association with Stresstech Oy, which provides solutions for manufacturing quality control. Their sensor, based on the Barkhausen noise effect, requires accurate positioning. The accuracy requirement admits a tool center point calibration problem if measurements are executed with an industrial robot. Multiple possibilities are available in the market for automatic tool center point calibration. Manufacturers provide customized calibrators to most robot types and tools. With the handmade sensors and multiple robot types that Stresstech uses, this would require great deal of labor. This thesis introduces a calibration method that is suitable for all robots which have two digital input ports free. It functions with the traditional method of using a light barrier to detect the tool in the robot coordinate system. However, this method utilizes two parallel light barriers to simultaneously measure and detect the center axis of the tool. Rotations about two axes are defined with the center axis. The last rotation about the Z-axis is calculated for tools that have different width of X- and Y-axes. The results indicate that this method is suitable for calibrating the geometric tool center point of a Barkhausen noise sensor. In the repeatability tests, a standard deviation inside robot repeatability was acquired. The Barkhausen noise signal was also evaluated after recalibration and the results indicate correct calibration. However, future studies should be conducted using a more accurate manipulator, since the method employs the robot itself as a measuring device.

Signaled two-way avoidance learning using electrical stimulation of the inferior colliculus as negative reinforcement: effects of visual and auditory cues as warning stimuli

The inferior colliculus is a primary relay for the processing of auditory information in the brainstem. The inferior colliculus is also part of the so-called brain aversion system as animals learn to switch off the electrical stimulation of this structure. The purpose of the present study was to determine whether associative learning occurs between aversion induced by electrical stimulation of the inferior colliculus and visual and auditory warning stimuli. Rats implanted with electrodes into the central nucleus of the inferior colliculus were placed inside an open-field and thresholds for the escape response to electrical stimulation of the inferior colliculus were determined. The rats were then placed inside a shuttle-box and submitted to a two-way avoidance paradigm. Electrical stimulation of the inferior colliculus at the escape threshold (98.12 ± 6.15 (A, peak-to-peak) was used as negative reinforcement and light or tone as the warning stimulus. Each session consisted of 50 trials and was divided into two segments of 25 trials in order to determine the learning rate of the animals during the sessions. The rats learned to avoid the inferior colliculus stimulation when light was used as the warning stimulus (13.25 ± 0.60 s and 8.63 ± 0.93 s for latencies and 12.5 ± 2.04 and 19.62 ± 1.65 for frequencies in the first and second halves of the sessions, respectively, P<0.01 in both cases). No significant changes in latencies (14.75 ± 1.63 and 12.75 ± 1.44 s) or frequencies of responses (8.75 ± 1.20 and 11.25 ± 1.13) were seen when tone was used as the warning stimulus (P>0.05 in both cases). Taken together, the present results suggest that rats learn to avoid the inferior colliculus stimulation when light is used as the warning stimulus. However, this learning process does not occur when the neutral stimulus used is an acoustic one. Electrical stimulation of the inferior colliculus may disturb the signal transmission of the stimulus to be conditioned from the inferior colliculus to higher brain structures such as amygdala

Effects of lead and/or zinc exposure during the second stage of rapid postnatal brain growth on delta-aminolevulinate dehydratase and negative geotaxis of suckling rats

Lead has been shown to produce cognitive and motor deficits in young rats that could be mediated, at least in part, by inhibition of the zinc-containing heme biosynthetic enzyme delta-aminolevulinate dehydratase (ALA-D). In the present study we investigated the effects of lead and/or zinc treatment during the second stage of rapid postnatal brain development on brain, kidney and blood ALA-D specific activity, as well as the negative geotaxis behavior of rats. Eight-day-old Wistar rats were injected intraperitoneally with saline, lead acetate (8 mg/kg) and/or zinc chloride (2 mg/kg) daily for five consecutive days. Twenty-four hours after treatment, ALA-D activity was determined in the absence and presence of DL-dithiothreitol (DTT). The negative geotaxis behavior was assessed in 9- to 13-day-old rats. Treatment with lead and/or zinc did not affect body, brain or kidney weights or brain- or kidney-to-body weight ratios of the animals. In spite of the absence of effect of any treatment on ALA-D specific activity in brain, kidney and blood, the reactivation index with DTT was higher in the groups treated with lead or lead + zinc than in the control group, in brain, kidney and blood (mean ± SEM; brain: 33.33 ± 4.34, 38.90 ± 8.24, 13.67 ± 3.41; kidney: 33.50 ± 2.97, 37.60 ± 2.67, 15.80 ± 2.66; blood: 63.95 ± 3.73, 56.43 ± 5.93, 31.07 ± 4.61, respectively, N = 9-11). The negative geotaxis response behavior was not affected by lead and/or zinc treatment. The results indicate that lead and/or zinc treatment during the second stage of rapid postnatal brain growth affected ALA-D, but zinc was not sufficient to protect the enzyme from the effects of lead in brain, kidney and blood.

Presence of the RHD pseudogene and the hybrid RHD-CE-Ds gene in Brazilians with the D-negative phenotype

The molecular basis for RHD pseudogene or RHDpsi is a 37-bp insertion in exon 4 of RHD. This insertion, found in two-thirds of D-negative Africans, appears to introduce a stop codon at position 210. The hybrid RHD-CE-Ds, where the 3' end of exon 3 and exons 4 to 8 are derived from RHCE, is associated with the VS+V- phenotype, and leads to a D-negative phenotype in people of African origin. We determined whether Brazilian blood donors of heterogeneous ethnic origin had RHDpsi and RHD-CE-Ds. DNA from 206 blood donors were tested for RHDpsi by a multiplex PCR that detects RHD, RHDpsi and the C and c alleles of RHCE. The RHD genotype was determined by comparison of size of amplified products associated with the RHD gene in both intron 4 and exon 10/3'-UTR. VS was determined by amplification of exon 5 of RHCE, and sequencing of PCR products was used to analyze C733G (Leu245Val). Twenty-two (11%) of the 206 D-negative Brazilians studied had the RHDpsi, 5 (2%) had the RHD-CE-Ds hybrid gene associated with the VS+V- phenotype, and 179 (87%) entirely lacked RHD. As expected, RHD was deleted in all the 50 individuals of Caucasian descent. Among the 156 individuals of African descent, 22 (14%) had inactive RHD and 3% had the RHD-CE-Ds hybrid gene. These data confirm that the inclusion of two different multiplex PCR for RHD is essential to test the D-negative Brazilian population in order to avoid false-positive typing of polytransfused patients and fetuses.