The non-palindromic adaptor-PCR method for the identification of the T-cell receptor genes of an interferon-gamma-secreting T-cell hybridomaspecific for trans-sialidase, an immunodominant Trypanosoma cruzi antigen

Cloning of the T-cell receptor genes is a critical step when generating T-cell receptor transgenic mice. Because T-cell receptor molecules are clonotypical, isolation of their genes requires reverse transcriptase-assisted PCR using primers specific for each different Valpha or Vß genes or by the screening of cDNA libraries generated from RNA obtained from each individual T-cell clone. Although feasible, these approaches are laborious and costly. The aim of the present study was to test the application of the non-palindromic adaptor-PCR method as an alternative to isolate the genes encoding the T-cell receptor of an antigen-specific T-cell hybridoma. For this purpose, we established hybridomas specific for trans-sialidase, an immunodominant Trypanosoma cruzi antigen. These T-cell hybridomas were characterized with regard to their ability to secrete interferon-gamma, IL-4, and IL-10 after stimulation with the antigen. A CD3+, CD4+, CD8- interferon-gamma-producing hybridoma was selected for the identification of the variable regions of the T-cell receptor by the non-palindromic adaptor-PCR method. Using this methodology, we were able to rapidly and efficiently determine the variable regions of both T-cell receptor chains. The results obtained by the non-palindromic adaptor-PCR method were confirmed by the isolation and sequencing of the complete cDNA genes and by the recognition with a specific antibody against the T-cell receptor variable ß chain. We conclude that the non-palindromic adaptor-PCR method can be a valuable tool for the identification of the T-cell receptor transcripts of T-cell hybridomas and may facilitate the generation of T-cell receptor transgenic mice.

Statistical analysis of event-related potential elicited by verb-complement merge in Brazilian Portuguese

An interesting fact about language cognition is that stimulation involving incongruence in the merge operation between verb and complement has often been related to a negative event-related potential (ERP) of augmented amplitude and latency of ca. 400 ms - the N400. Using an automatic ERP latency and amplitude estimator to facilitate the recognition of waves with a low signal-to-noise ratio, the objective of the present study was to study the N400 statistically in 24 volunteers. Stimulation consisted of 80 experimental sentences (40 congruous and 40 incongruous), generated in Brazilian Portuguese, involving two distinct local verb-argument combinations (nominal object and pronominal object series). For each volunteer, the EEG was simultaneously acquired at 20 derivations, topographically localized according to the 10-20 International System. A computerized routine for automatic N400-peak marking (based on the ascendant zero-cross of the first waveform derivative) was applied to the estimated individual ERP waveform for congruous and incongruous sentences in both series for all ERP topographic derivations. Peak-to-peak N400 amplitude was significantly augmented (P < 0.05; one-sided Wilcoxon signed-rank test) due to incongruence in derivations F3, T3, C3, Cz, T5, P3, Pz, and P4 for nominal object series and in P3, Pz and P4 for pronominal object series. The results also indicated high inter-individual variability in ERP waveforms, suggesting that the usual procedure of grand averaging might not be considered a generally adequate approach. Hence, signal processing statistical techniques should be applied in neurolinguistic ERP studies allowing waveform analysis with low signal-to-noise ratio.

A carbohydrate-based mechanism of species recognition in sea urchin fertilization

In the present review, we describe a systematic study of the sulfated polysaccharides from marine invertebrates, which led to the discovery of a carbohydrate-based mechanism of sperm-egg recognition during sea urchin fertilization. We have described unique polymers present in these organisms, especially sulfated fucose-rich compounds found in the egg jelly coat of sea urchins. The polysaccharides have simple, linear structures consisting of repeating units of oligosaccharides. They differ among the various species of sea urchins in specific patterns of sulfation and/or position of the glycosidic linkage within their repeating units. These polysaccharides show species specificity in inducing the acrosome reaction in sea urchin sperm, providing a clear-cut example of a signal transduction event regulated by sulfated polysaccharides. This distinct carbohydrate-mediated mechanism of sperm-egg recognition coexists with the bindin-protein system. Possibly, the genes involved in the biosynthesis of these sulfated fucans did not evolve in concordance with evolutionary distance but underwent a dramatic change near the tip of the Strongylocentrotid tree. Overall, we established a direct causal link between the molecular structure of a sulfated polysaccharide and a cellular physiological event - the induction of the sperm acrosome reaction in sea urchins. Small structural changes modulate an entire system of sperm-egg recognition and species-specific fertilization in sea urchins. We demonstrated that sulfated polysaccharides - in addition to their known function in cell proliferation, development, coagulation, and viral infection - mediate fertilization, and respond to evolutionary mechanisms that lead to species diversity.

Serotonergic modulation of face-emotion recognition

Facial expressions of basic emotions have been widely used to investigate the neural substrates of emotion processing, but little is known about the exact meaning of subjective changes provoked by perceiving facial expressions. Our assumption was that fearful faces would be related to the processing of potential threats, whereas angry faces would be related to the processing of proximal threats. Experimental studies have suggested that serotonin modulates the brain processes underlying defensive responses to environmental threats, facilitating risk assessment behavior elicited by potential threats and inhibiting fight or flight responses to proximal threats. In order to test these predictions about the relationship between fearful and angry faces and defensive behaviors, we carried out a review of the literature about the effects of pharmacological probes that affect 5-HT-mediated neurotransmission on the perception of emotional faces. The hypothesis that angry faces would be processed as a proximal threat and that, as a consequence, their recognition would be impaired by an increase in 5-HT function was not supported by the results reviewed. In contrast, most of the studies that evaluated the behavioral effects of serotonin challenges showed that increased 5-HT neurotransmission facilitates the recognition of fearful faces, whereas its decrease impairs the same performance. These results agree with the hypothesis that fearful faces are processed as potential threats and that 5-HT enhances this brain processing.

A novel polar-based human face recognition computational model

Motivated by a recently proposed biologically inspired face recognition approach, we investigated the relation between human behavior and a computational model based on Fourier-Bessel (FB) spatial patterns. We measured human recognition performance of FB filtered face images using an 8-alternative forced-choice method. Test stimuli were generated by converting the images from the spatial to the FB domain, filtering the resulting coefficients with a band-pass filter, and finally taking the inverse FB transformation of the filtered coefficients. The performance of the computational models was tested using a simulation of the psychophysical experiment. In the FB model, face images were first filtered by simulated V1- type neurons and later analyzed globally for their content of FB components. In general, there was a higher human contrast sensitivity to radially than to angularly filtered images, but both functions peaked at the 11.3-16 frequency interval. The FB-based model presented similar behavior with regard to peak position and relative sensitivity, but had a wider frequency band width and a narrower response range. The response pattern of two alternative models, based on local FB analysis and on raw luminance, strongly diverged from the human behavior patterns. These results suggest that human performance can be constrained by the type of information conveyed by polar patterns, and consequently that humans might use FB-like spatial patterns in face processing.

Long-term social recognition memory in adult male rats: factor analysis of the social and non-social behaviors

A modified version of the intruder-resident paradigm was used to investigate if social recognition memory lasts at least 24 h. One hundred and forty-six adult male Wistar rats were used. Independent groups of rats were exposed to an intruder for 0.083, 0.5, 2, 24, or 168 h and tested 24 h after the first encounter with the familiar or a different conspecific. Factor analysis was employed to identify associations between behaviors and treatments. Resident rats exhibited a 24-h social recognition memory, as indicated by a 3- to 5-fold decrease in social behaviors in the second encounter with the same conspecific compared to those observed for a different conspecific, when the duration of the first encounter was 2 h or longer. It was possible to distinguish between two different categories of social behaviors and their expression depended on the duration of the first encounter. Sniffing the anogenital area (49.9% of the social behaviors), sniffing the body (17.9%), sniffing the head (3%), and following the conspecific (3.1%), exhibited mostly by resident rats, characterized social investigation and revealed long-term social recognition memory. However, dominance (23.8%) and mild aggression (2.3%), exhibited by both resident and intruders, characterized social agonistic behaviors and were not affected by memory. Differently, sniffing the environment (76.8% of the non-social behaviors) and rearing (14.3%), both exhibited mostly by adult intruder rats, characterized non-social behaviors. Together, these results show that social recognition memory in rats may last at least 24 h after a 2-h or longer exposure to the conspecific.

Effect of metabolic syndrome and of its individual components on renal function of patients with type 2 diabetes mellitus

The objective of this study was to evaluate the effect of metabolic syndrome (MetS) and its individual components on the renal function of patients with type 2 diabetes mellitus (DM). A cross-sectional study was performed in 842 type 2 DM patients. A clinical and laboratory evaluation, including estimated glomerular filtration rate (eGFR) calculated by the modification of diet in renal disease formula, was performed. MetS was defined according to National Cholesterol Education Program - Adult Treatment Panel III criteria. Mean patient age was 57.9 ± 10.1 years and 313 (37.2%) patients were males. MetS was detected in 662 (78.6%) patients. A progressive reduction in eGFR was observed as the number of individual MetS components increased (one: 98.2 ± 30.8; two: 92.9 ± 28.1; three: 84.0 ± 25.1; four: 83.8 ± 28.5, and five: 79.0 ± 23.0; P < 0.001). MetS increased the risk for low eGFR (<60 mL·min-1·1.73 (m²)-1) 2.82-fold (95%CI = 1.55-5.12, P < 0.001). Hypertension (OR = 2.2, 95%CI = 1.39-3.49, P = 0.001) and hypertriglyceridemia (OR = 1.62, 95%CI = 1.19-2.20, P = 0.002) were the individual components with the strongest associations with low eGFR. In conclusion, there is an association between MetS and the reduction of eGFR in patients with type 2 DM, with hypertension and hypertriglyceridemia being the most important contributors in this sample. Interventional studies should be conducted to determine if treatment of MetS can prevent renal failure in type 2 DM patients.

Interaction between affordance and handedness recognition: a chronometric study

The visualization of tools and manipulable objects activates motor-related areas in the cortex, facilitating possible actions toward them. This pattern of activity may underlie the phenomenon of object affordance. Some cortical motor neurons are also covertly activated during the recognition of body parts such as hands. One hypothesis is that different subpopulations of motor neurons in the frontal cortex are activated in each motor program; for example, canonical neurons in the premotor cortex are responsible for the affordance of visual objects, while mirror neurons support motor imagery triggered during handedness recognition. However, the question remains whether these subpopulations work independently. This hypothesis can be tested with a manual reaction time (MRT) task with a priming paradigm to evaluate whether the view of a manipulable object interferes with the motor imagery of the subject's hand. The MRT provides a measure of the course of information processing in the brain and allows indirect evaluation of cognitive processes. Our results suggest that canonical and mirror neurons work together to create a motor plan involving hand movements to facilitate successful object manipulation.

Pertussis: infection, vaccination and gene polymorphisms

Pertussis or whooping cough is a human respiratory tract infection and a vaccine-preventable disease that is caused by Bordetella pertussis bacteria. Pertussis vaccination has been part of the Finnish national vaccine program since 1952. Despite extensive vaccinations, the incidence of pertussis has increased in many countries during the last decades. Large epidemics have been observed also in countries with high vaccine coverage. Inter-individual variation in immune responses is always encountered after vaccination. Low vaccine responses may cause vulnerability to pertussis even straight after vaccination. Reasons for low responses are not fully understood. The innate immune system is responsible for the initial recognition of pathogens and vaccine antigens. The role of innate immunity on pertussis immunity has not been thoroughly investigated. Mannose-binding lectin (MBL) and toll-like receptor 4 (TLR4) are important molecules of the innate immune system and in the recognition of pathogens. Cytokines form a signaling network that have a notable role in immune responses after infections as well as after vaccinations. Single nucleotide polymorphism (SNP) is common in genes encoding these molecules and the polymorphisms have been reported to affect vaccine response after viral and bacterial vaccines. This study investigated the gene polymorphisms of MBL2, TLR4 and interleukin (IL)-10 promoter and their association with vaccine responses after acellular pertussis (aP) vaccination in Finnish adolescents and infants. Cell-mediated immune responses were investigated ten years after the previous pertussis vaccinations in young adults. In addition, the role of MBL deficiency in pertussis infection susceptibility was evaluated. The results of this study show that subjects with TLR4 polymorphism had lower antibody production and persistence after aP vaccination compared with normal allele. A specific SNP in the TLR4 gene was associated with decreased antibody responses and persistence in adolescents after aP booster vaccination. Cell-mediated immune responses were partly detected ten years after the previous vaccination; booster vaccine clearly enhanced the responses. In addition, subjects with IL-10 polymorphism had altered cell-mediated immune responses. MBL deficiency was found to be more frequent in pertussis patients than healthy controls but the polymorphism of MBL2 was not associated with antibody responses after acellular pertussis vaccination. The novel finding of this study was that genetic variation in the innate immune system seems to play a role in altered pertussis vaccine responses as well as in pertussis infection. These new findings enlighten the mechanisms behind the low responses after pertussis vaccination and help to predict risk factors related to this phenomenon.

Comportamento individual de plantas originadas de sementes com diferentes níveis de qualidade fisiológica em populações de arroz irrigado

Com o objetivo de avaliar o comportamento individual de plantas originadas de sementes de alto e baixo vigor em populações de arroz irrigado (Oryza sativa L.), constituídas de diferentes combinações de distribuição ao longo da linha de semeadura, foi conduzido este estudo. O experimento constou de diferentes combinações de distribuição de sementes de arroz, cv. BR-IRGA 410, oriundas de lotes de alto e baixo vigor, semeadas em ambiente controlado e, após, transplantadas ao campo para garantir o estande de plantas e o arranjo da semeadura. Para tal, foram avaliados altura, matéria seca da planta e número de perfilhos aos 60, 102 e 134 dias após transplante ao campo e número de panículas por planta, rendimento de grãos e índice de colheita, ao final do ciclo. As plantas originadas de sementes de alto vigor apresentaram desempenho superior para todas as características avaliadas, independentemente do sistema de distribuição das plantas ao longo da linha de semeadura. As plantas originadas de sementes de alto vigor apresentaram rendimento médio de grãos superior ao das plantas originadas de sementes de baixo vigor. Plantas de arroz originadas de sementes de alto vigor não apresentam desempenho dominante sobre as plantas adjacentes de baixo vigor na linha de semeadura.

Desempenho individual e de populações de plantas de arroz híbrido em função da qualidade fisiológica das sementes

O trabalho teve por objetivo avaliar o comportamento individual e de populações de plantas de arroz híbrido originadas de sementes de alto e de baixo vigor, dentro de populações que foram constituídas de diferentes combinações de distribuição dessas plantas ao longo da linha de semeadura. O experimento foi conduzido em condições de campo na Universidade Federal de Pelotas, Capão do Leão/RS, constando de cinco tratamentos (proporções de 0, 25, 50, 75 e 100% de plantas originadas de sementes de alto vigor na população). Para tanto, utilizou-se sementes de arroz do híbrido Avaxi da empresa RiceTec®, originadas de dois lotes devidamente caracterizados como alto e baixo vigor. A semeadura de cada lote foi realizada em bandejas distintas com solo como substrato. Após a emergência das plântulas foi realizado um desbaste nas bandejas, retirando as plantas mais precoces no lote de baixo vigor e as plântulas mais tardias no lote de alto vigor, com o propósito de utilizar as plântulas emergidas no dia de maior freqüência de emergência em cada um dos níveis de vigor. O transplante das plantas foi realizado aos 20 dias após a semeadura. Utilizou-se o delineamento experimental de blocos ao acaso com 3 repetições. Avaliou-se: a estatura das plantas aos 60 Dias Após Transplante (DAT), área foliar, número de perfilhos aos 60 DAT, número de panículas por planta aos 115 DAT, número de grãos por panícula, número de grãos por planta, e rendimento de grãos por planta. As plantas originadas de sementes de alto vigor apresentaram desempenho superior às plantas provenientes de sementes de baixo vigor para as características avaliadas, independente do sistema de distribuição das plantas ao longo da linha de semeadura. O uso de sementes de alto vigor no estabelecimento de comunidades de arroz híbrido proporciona acréscimos superiores a 30% no rendimento de grãos.

Individual Crimes or a Sociological Phenomenon: A Critical discourse analysis of the 2014 Royal Canadian Mounted Police National Operational Overview on missing and murdered Indigenous women in Canada

Pro gradu -tutkielma käsittelee Kanadan poliisivoimien, Royal Canadian Mounted Police (RCMP), vuoden 2014 operationaalista raporttia Kanadan kadonneista ja murhatuista alkuperäiskansojen naisista. Heitä katoaa ja murhataan Kanadassa suhteessa huomattavasti enemmän kuin muita naisia. Aihe on hyvin ajankohtainen sekä kriittinen ja siihen on paikallisten organisaatioiden lisäksi kiinnittänyt huomiota mm. Amnesty International ja Yhdistyneet Kansakunnat. Tilannetta pidetään jo ihmisoikeuskriisinä ja sen puolesta puhuminen voidaan nähdä osana laajempaa kansannousua alkuperäiskansojen oikeuksien puolesta. Kanadan hallitusta sekä poliisivoimia on syytetty välinpitämättömyydestä ja rasismista, poliiseja jopa väkivallasta alkuperäiskansojen naisia kohtaan. Kanadan hallitus ei myönnä ongelman olevan sosiologinen ilmiö, vaan pääministerin mukaan naisten katoamiset ja murhat ovat yksittäisiä rikoksia. Tilanteen taustalla on laajempi ongelma, joka juontaa juurensa kolonialismista, alkuperäiskansojen pakkokoulutuksesta ja heidän kulttuurinsa sekä identiteettinsä tuhoamisesta. Ennakkoluulot ja stereotypiat elävät yhä vahvana luoden heikon aseman alkuperäiskansojen edustajille ja tässä tapauksessa erityisesti naisille, joita myös usein syytetään omista ongelmistaan. Tutkielma selvittää, onko tämä operationaalinen raportti eriarvoisuutta, stereotypioita sekä ennakkoluuloja ylläpitävä ja tukeeko se näkemystä sosiologisesta ilmiöstä. Poliisivoimat ovat yhteiskunnassa vaikutusvaltaisessa asemassa ja voivat edesauttaa, jatkaa ja voimistaa syrjivää ajattelua, käytöstä ja viestintää. Vaikka he myös ajavat alkuperäiskansojen etuja, tulee heidän toimintaansa ja täten tätä kyseistä raporttia tarkastella kriittisesti. Tutkielma keskittyy kriittiseen diskurssianalyysiin (Fairclough 1995, Wodak & Meyer 2009). Raportin teksti ei ole avoimesti diskriminoivaa ja keskittyy tilastojen ja lukujen raportointiin sekä vertailuun, luoden samalla kuvan asianomaisista sekä tilanteesta yleisesti. Makroanalyysi (van Dijk 1977, 1980) mahdollistaa korkeamman tason ajatusten ja ideoiden esille tuonnin paljastaen ns. rivien välistä löytyvän viestinnän. Näistä makroanalyysin tuloksista etsitään erilaisia diskurssistrategioita (Reisigl & Wodak 2001), joiden avulla luodaan mm. kuva positiivisesta itsestä ja negatiivisesta toisesta. Näitä tuloksia tuetaan lyhyellä multimodaalisella analyysillä, joka keskittyy lähinnä raportin kahteen kuvaan (Kress & van Leeuwen 2006). Lopuksi tuloksista keskustellaan suhteessa yhteiskunnan valtasuhteisiin (Foucault 1976), rasismiin ja seksismiin. Analyysin perusteella raportti on selkeästi diskriminoiva. Se tuo esiin vanhoja stereotypioita ja ennakkoluuloja, esittää tilanteen alkuperäiskansojen sisäisenä ongelmana, jättää mainitsematta tilanteen taustat ja luo hyvin positiivisen kuvan poliisivoimista (RCMP). Tällä tavoin se ylläpitää yhteiskunnan epätasapainoisia valtasuhteita ja alkuperäiskansojen naisten heikompaa ja marginalisoitua asemaa. Raportti todistaa kuinka syvällä Kanadan yhteiskunnassa jopa institutionaalisella tasolla nämä ajatukset piilevät, sillä diskriminaatio ulottuu tapausten syistä niiden käsittelyyn ja raportointiin. Tilanne on huolestuttava ja voidaan nähdä sosiologisena ilmiönä. Poliisivoimien tulisi lisätä sisäistä koulutusta asian suhteen sekä hallituksen tutkia tätä ilmiönä ja paneutua sen selvittämiseen laaja-alaisesti ja läpinäkyvästi.