This grounded theory study explored the influences that surrounded decision making for people with a minimal requirement for care, who entered a care home with registered nurses (RN’s) when their needs could be met in an alternative environment. The study was undertaken in a geographical area in England, which has in excess of 3,500 care home beds staffed with RNs. In May 2011, there were reported to be 183 residents with minimal care needs, resident in a care home with RNs. The study utilized a grounded theory methodology as described by Glaser and Strauss with influences from Charmaz. The initial sampling was purposive progressing to theoretical. Interviews were conducted with twelve care home residents responsible for funding their own care home fees, assessed as not requiring care by RNs. Data analysis was in accordance with the principles of grounded theory and identified two main categories. The first category, entitled “choosing the path” focused on the decision making process which described a perceived paucity of help and support received by participants and their families in identifying and exploring the available options. The second category termed “settling in” related to the process of adaptation to the new environment. It appeared that participants with greater control over the decision making process found it easier to settle into the home, conversely those with the least control found it more difficult to adapt. These two categories linked to form the emerging theory of “crossing the bridge” from independent living to becoming a care home resident. It is important health and social care practitioners use appropriate evidence based knowledge when assisting older people and their families to consider available options. The findings of the study add to the body of evidence concerning the factors that influence people when considering their future care and the importance of informed decision-making.

Learning can be explicit or implicit. Explicit learning takes place intentionally, in the presence of factual task-relevant knowledge; whereas implicit learning takes place unintentionally, without concurrent acquisition of knowledge about task performance. The relative benefits of implicit learning have been well investigated within healthy populations. Research consistently demonstrates that skills learnt implicitly are more likely to be retained, and are more robust under secondary task load. However, study protocols tend to involve laboratory based activities, which do not take into account the complexities of motor learning in natural settings. Direct transferability of the findings to stroke rehabilitation is therefore questionable. Two factors in explicit and implicit learning are the concepts of attentional capacity and attentional focus. Attentional capacity refers to the ability to attend to and process incoming information, whereas attentional focus refers to the location of attention in relation to specific aspects of the task being performed. Theories propose that focussing on specific movements (internal focus) may actually constrain or interfere with automatic control processes that would normally regulate movement, whereas if attention is focussed towards the movement effect (external focus) the motor system is able to more naturally self-organize, resulting in more effective performance, and learning. An internal focus of attention is therefore allied to explicit learning; whilst an external focus of attention is allied to implicit learning. This research aimed to improve understanding of explicit and implicit learning within early gait rehabilitation post stroke; primarily through the development and testing of explicit and implicit models of learning interventions. It has comprised three phases; a review of the literature; an observational study to gain insight into current practice; and a feasibility study to test the ability of therapists to deliver interventions with a bias towards either an explicit or implicit approach. Therapists were found to favour the use of explicit techniques; internally focussed instructions and feedback statements were used in high quantities. Practice therefore appeared to be at odds with current evidence; albeit primarily from healthy populations. Guidance for the delivery of explicit and implicit learning models in clinical practice was developed, and then tested in a feasibility study. Therapists demonstrated the ability to change their practice to bias either explicit or implicit learning; both approaches were found to be acceptable to patients and therapists. Recommendations are made on the content and evaluation of explicit and implicit learning models in future research, and specifically, in a Phase II pilot study.

The ability to learn and work in a multidisciplinary team is crucial to the delivery of healthcare that is ‘fit for purpose’ in the twenty-first century. Consequently, interprofessional education (IPE) is a mandatory requirement for pre-registration training in health and social care. However, the evidence base for the effectiveness of such programmes remains limited. This evaluative study aims to explore the potential influence of IPE at pre-registration level, using podiatry as an exemplar. In Phase One, key informant interviews were held with IPE curriculum developers from higher education institutions (HEIs) (n=8) in the United Kingdom. Verbatim transcripts of these interviews were analysed using elements of grounded theory. In Phase Two, using the qualitative results as the anchor, a survey was constructed and administered to pre-registration health and social work students in a number of parallel IPE initiatives (Sample 1: n=1151; Sample 2: n=1060), involving 6 and 5 HEIs respectively. The findings of the key informant interviews and SPSS survey analysis were subsequently combined to inform the results of the research. The findings revealed an absence of educational theory underpinning the IPE curriculum development process. In addition there were issues concerning institutional commitment to IPE, attitudes towards IPE, and inconsistencies in the approach to curriculum delivery, all of which militated against successful implementation. The results illuminated that students learn about other health and social care professions that are represented in their IPE group. An appropriate time to introduce IPE into pre-registration programmes was found to be the latter part of the academic year for students studying first year modules. Students’ readiness for interprofessional learning was found to be enhanced if they were inducted to a coherent IPE initiative with consistent approaches to learning, teaching, assessment and evaluation across programmes. However, attitude differences between professional groups emerged where students with a strong sense of professional identity and roles were less ready to engage in interprofessional learning. This study indicates that the current method of implementing IPE in pre-registration health and social care programmes is less than ideal. The development and delivery of the IPE curriculum appears to be flawed. Explanations for the findings are explored and the implications for practice and future research are considered.

SOUZA, Isabel Cristina da Cósta. Caracterização bioquímica de calos de espécies laticíferas em resposta ao estresse salino e análise da transcrição de osmotinas. 2015. 107 f. Dissertação (mestrado em bioquímica)- Universidade Federal do Ceará, Fortaleza-CE, 2015. / Submitted by Elineudson Ribeiro (elineudsonr@gmail.com) on 2016-03-22T17:57:48Z No. of bitstreams: 1 2016_dis_iccsouza.pdf: 2040395 bytes, checksum: f08a9aac46cd6e718bf109abee257b2f (MD5) / Approved for entry into archive by José Jairo Viana de Sousa (jairo@ufc.br) on 2016-04-28T22:22:53Z (GMT) No. of bitstreams: 1 2016_dis_iccsouza.pdf: 2040395 bytes, checksum: f08a9aac46cd6e718bf109abee257b2f (MD5) / Made available in DSpace on 2016-04-28T22:22:53Z (GMT). No. of bitstreams: 1 2016_dis_iccsouza.pdf: 2040395 bytes, checksum: f08a9aac46cd6e718bf109abee257b2f (MD5) Previous issue date: 2015 / Calotropis procera e Cryptostegia grandiflora are laticiferous plants. It was found osmotin protein. The literature shows that the osmotinas are associated to plant defence mechanisms in situations of biotic or abiotic stress. However, there are still several inconsistencies in this hypothesis. In this context, it was used in vitro tissue culture techniques as model to assist in the understanding of how the C. procera and C. grandiflora callus cells respond to salt stress in biochemical terms, and whether the transcripts level for osmotin has raised in response to exposure to NaCl. It was added NaCl to the culture medium of Murashige e Skoog (MS) in increasing concentrations (0, 20, 40, 60 e 80 mM). The results show that callus treated with 80 mM NaCl have reduced the growth and the humidity percentage of respectively 33% and 10% in C. procera and 83% and 39% in C. grandiflora compared to the control treatment callus. Under the same conditions, it was seen an increase in ions concentrations of Na+ and Cl-, 98.9% and 98% in C. procera and 98.8% and 96% in C. grandiflora respectively. It was also seen a reduction in K+ level in callus treated with 80 mM NaCl, 43% in C. procera and 18% in C. Grandiflora, when compared to the control. The callus treated with 80 mM NaCl, showed a tendency of the proline accumulation and soluble sugars, increasing 26% and 37% in C. procera callus and 55.4% and 45% in C. grandiflora callus, respectively, when compared to control conditions. The increase in the activity of enzymes that break H2O2 has been observed in C. grandiflora callus under the salt stress, suggesting a possible oxidative damage, this increase was 73% in the ascorbate peroxidase activity and 62% in guaiacol peroxidase activity when compared to the activity of enzymes of the control callus with the treaty in 80 mM NaCl. None connection was seen between changes in the activity of the enzymes of the oxidative system and the salt stress in C. procera callus. It was evaluated the behaviour of osmotin in the osmotin transcription at 0, 2, 12, 24, 48 hours and at 4, 7, 14, 28 days of callus contact under stress. The osmotin transcripts were observed from 12 hours of contact of callus in 80mM NaCl in both species. However, it was not found osmotin by electrophoresis assays, dot blotting and mass spectrometry in the protein extracts of C. procera and C. grandiflora callus grown in control conditions and in 80 mM NaCl. Thus, the salt stress evaluation using in vitro cell model study was effective, providing cellular behaviour information for these two laticifers plants species showing their physiological, biochemical and molecular changes. The results suggest that the induced salt stress has favoured the increase of osmotin gene expression in both cases and suggests a possible relationship between osmotin of these species with the protection to salinity conditions. The failure to detect the proteins corresponding to genes provides the conception of several new hypotheses to be validated. / Calotropis procera e Cryptostegia grandiflora são plantas laticíferas. Em seus fluidos laticíferos, foram encontradas proteínas do tipo osmotina. A literatura reporta que osmotinas são proteínas relacionadas com mecanismos de defesa vegetal em situações de estresse biótico e/ou abiótico. Entretanto, ainda há várias inconsistências nessa afirmação. Nesse contexto, técnicas in vitro de cultura de tecidos vegetais foram aplicadas como modelo para auxiliar na compreensão de como as células de calos de C. procera e C. grandiflora respondem ao estresse salino, em termos bioquímicos, e se o nível de transcritos para a osmotina seria aumentado em resposta à exposição a NaCl. Para indução desse estresse, NaCl foi adicionado à formulação nutritiva de Murashige e Skoog (MS), em concentrações crescentes (0, 20, 40, 60 e 80 mM). Os resultados mostram que os calos cultivados com NaCl a 80 mM tiveram o crescimento e o teor de umidade reduzidos, respectivamente, em 33% e 10%, em C. procera e de 83% e 39%, em C. grandiflora, em comparação ao seu tratamento controle. Nessas mesmas condições, foi observado um aumento nas concentrações dos íons Na+ e Cl- de, respectivamente, 98,9% e 98%, em C. procera, e de 98,8% e 96%, em C. grandiflora. Foi também observada diminuição no teor de K+ nos calos tratados com NaCl a 80 mM. Essa redução foi de 43%, em C. procera, e de 18% em C. grandiflora, quando comparado ao tratamento controle. Os calos tratados com NaCl a 80 mM, apresentaram uma tendência de acúmulo de prolina e açúcares solúveis, alcançando, respectivamente valores, 26% e 37% maiores em calos de C. procera, e 55,4% e 45% maiores, em calos de C. grandiflora, que aqueles em condições controle. O aumento na atividade das enzimas que degradam H2O2 foi observado em calos de C. grandiflora submetidos a estresse salino, sugerindo um possível dano oxidativo. Esse aumento foi de 73%, para a ascorbato peroxidase, e de 62% para a peroxidase do guaiacol, nos calos tratados com NaCl a 80 mM, em relação ao controle. Não foi observada qualquer alteração significante na atividade das enzimas do sistema antioxidativo em razão do estresse salino em calos de C. procera. Em relação à transcrição da osmotina, foi avaliado o perfil de seus transcritos nos intervalos de tempo de 0, 2, 12, 24, 48 horas e de 4, 7, 14 e 28 dias sob estresse. Os transcritos de osmotina foram observados a partir de 12 horas de contato dos calos com NaCl a 80 mM, em ambas as espécies. Contudo, nos extratos proteicos dos calos de C. procera e C. grandiflora cultivados em condições controle e de 80 mM de NaCl, não foi detectada a presença da proteína osmotina quando avaliado pelos ensaios de eletroforese, Dot blotting e espectrometria de massas. Assim, a avaliação do estresse salino utilizando como modelo de estudo células in vitro foi eficiente, fornecendo informações do comportamento celular de duas espécies laticíferas, mostrando suas alterações fisiológicas, bioquímicas e moleculares. Os resultados sugerem que o estresse salino favoreceu o aumento da transcrição do gene da osmotina em calos das duas espécies em estudo e permite propor uma possível relação das osmotinas dessas espécies com a tolerância à salinidade. A falha em detectar as proteínas correspondentes aos genes propicia a concepção de várias novas hipóteses a serem validadas.

Dentre as oito espécies do gênero Listeria, a única transmitida por alimentos, patogênica para humanos, é a Listeria monocytogenes. De grande preocupação para a indústria alimentícia, este microrganismo pode ser encontrado em diversas superfícies de plantas processadoras de alimentos, sendo capaz de se aderir e formar persistentes biofilmes. No presente trabalho, foi estudada a influência da concentração de glicose e do tempo de incubação na formação de biofilme por uma cepa de L. monocytogenes, sorotipo 1/2a, cultivada em três diferentes temperaturas, 7°C, 25°C e 37°C, através de planejamento experimental utilizando a Metodologia de Superfície de Resposta. As duas variáveis testadas, para os intervalos estudados, foram significativas (p<0,05) na formação de biofilme somente na temperatura de 37°C e, tanto concentrações de glicose tendendo a zero combinadas com tempos de incubação próximos a 26 h, como maiores concentrações de glicose combinadas com menores tempos de incubação, dentro da faixa de estudo aplicada, representaram boas condições para a formação de biofilme. Foi também realizada a análise transcricional de genes relacionados à formação de biofilme e virulência em duas cepas de L. monocytogenes, sorotipos 1/2a e 4b, cultivadas a 7°C e 37°C (condição controle), pela técnica de PCR quantitativo em tempo real (RT-qPCR). Para ambas as cepas, os genes sigB, prfA, luxS, sufS, sufU, ltrC e flaA apresentaram transcrição significativamente aumentada (p<0,05) a 7°C, em comparação aos resultados para a condição de cultivo a 37°C, enquanto que o gene hly não variou significativamente a sua transcrição entre as duas temperaturas. O gene actA foi mais transcrito a 7°C para a cepa denominada 55, do sorotipo 1/2a, enquanto que não variou significativamente a sua transcrição entre as duas condições de crescimento para a cepa denominada 47, do sorotipo 4b. No entanto, para a cepa 55, o gene degU não demonstrou diferença estatisticamente significativa entre seus níveis de transcrição nas duas temperaturas estudadas, mas para a cepa 47, apresentou transcrição significativamente aumentada a 7°C. Interessantemente, a presença do gene agrA não foi detectada na cepa 47, e os seus níveis de transcrição foram menores a 7°C para a cepa 55. Estes resultados demonstram que os dois sorotipos estudados, responsáveis por muitos dos casos humanos de listeriose, apresentam mecanismos moleculares diferentes, nas temperaturas de 7°C e 37°C. / Among the eight species of genus Listeria, the only transmitted by food, pathogenic for humans, is Listeria monocytogenes. Of great concern to the food industry, this microorganism can be found in various areas of food processing plants, being able to adhere and form persistent biofilms. In the present work, the influence of glucose concentration and incubation time on biofilm formation by a strain of L. monocytogenes, serotype 1/2a, grown in three different temperatures, 7°C, 25°C and 37°C, have been studied, through an experimental design using the Response Surface Methodology. The two variables tested, for the studied intervals, were significant (p<0,05) in the biofilm formation only at a temperature of 37°C and, both glucose concentrations tending to zero combined with incubation times near to 26 h, and higher glucose concentrations combined with lower incubation times, within the applied range study, represented good conditions for biofilm formation. Transcriptional analysis of genes related to biofilm formation and virulence in two strains of L. monocytogenes, serotypes 1/2a and 4b, grown at 7°C and 37°C (control condition), was also performed, by real-time quantitative PCR (RT-qPCR). For both strains, sigB, prfA, luxS, sufS, sufU, ltrC and flaA genes showed significantly increased transcription (p<0,05) at 7°C, in comparison with results for the growth condition at 37°C, whereas hly gene did not varied significantly its transcription between the two temperatures. The actA gene was more transcribed at 7°C for the 55 strain, serotype 1/2a, while did not varied significantly its transcription between the two growth conditions for the 47 strain, serotype 4b. However, for 55 strain, the degU gene did not showed statistically significant difference for its transcription levels between the two studied temperatures, but for 47 strain, presented significantly increased transcription at 7°C. Interestingly, the presence of agrA gene was not detected in 47 strain, and its transcription levels were lower at 7°C for 55 strain. These results demonstrate that the two studied serotypes, responsible for many of the human listeriosis cases, have different molecular mechanisms, at temperatures of 7 and 37°C.

Made available in DSpace on 2014-06-11T19:25:37Z (GMT). No. of bitstreams: 0 Previous issue date: 2010-10-29Bitstream added on 2014-06-13T20:33:23Z : No. of bitstreams: 1 silva_ds_me_araca.pdf: 824748 bytes, checksum: a5191c74f89f8870aa31a3483beae50a (MD5) / O tumor venéreo transmissível canino (TVTC) é uma neoplasia transmitida entre cães saudáveis pelo contato direto de pele e/ou mucosas lesionadas. Face aos escassos estudos relacionados aos eventos celulares envolvidos nas fases de crescimento do TVTC, o presente estudo teve por objetivo identificar a presença do gene TP53 e o RNAm referente a proteína codificada, além de detectar a expressão das proteínas p53, Bcl-2 e p63 em cortes histológicos de 13 amostras de TVTC. Com relação à evolução da neoplasia, 46% das amostras foram consideradas em fase de progressão e 54% no estágio de regressão. Foram utilizadas as técnicas de hibridização in situ (ISH) e RT-PCR in situ, que demonstrou a presença do DNA homólogo ao TP53 e seu respectivo RNAm em 92,30% das amostras. A expressão das proteínas p53, p63 e Bcl-2 foram detectadas em 50%, 70% e 100% das amostras, respectivamente. A p63 foi expressa de forma evidente nas amostras em regressão, porém a p53 e a Bcl-2 não apresentaram relação com o estágio evolutivo do tumor e provavelmente não podem ser analisados como fatores de prognóstico do TVTC. Observou-se, nesse estudo que, através das técnicas de ISH e RT-PCR in situ foi possível detectar o DNA do TP53 e seus transcritos, porém esse fato não significou a transcrição da p53, devido aos baixos níveis de expressão nas análises quantitativa e qualitativa nas amostras de TVTC / The canine transmissible venereal tumor (CTVT) is transmitted by direct contact of skin or mucosal presenting lesions. In fact, few reports have been found describing the cellular immune response related to the evolution of the tumor. The objective of this study was to identify the TP53 gen and its transcription in CTVT in different stages of evolution, collected from dogs (N=13) examined at veterinary school, UNESP, Aracatuba, SP, Brasil. In addition, it was also evaluated the expression of p53, p63 and Bcl-2 in histological sections by the use of immunohistochemystry assay. The p53, p63 and Bcl-2 were evident in 50, 70 and 100% of analyzed samples. Regarding to tumor evolution, 6 out of 13 were considered in a progressive stage (46%), was list 7 out of 13 were classified in a regressive stage (54%). The use of in situ hybridization and reverse transcriptase polymerase chain reaction in situ (RT-PCR), revealed that TP53 was present in all samples and P63 was more expressed than p53. Take all results together, the real role of those marker are extremely important to understand the biological behavior and to improve therapeutic procedures for CTVT

Objective The aim of this study was to examine the effects of selected routine nursing interventions - endotracheal suctioning and manual ventilation (ETSMV), log-rolling, eye care, mouth care and washing - on the intracranial pressure (ICP) in children with traumatic brain injury. Design Prospective observational study over three years. Setting Single tertiary paediatric intensive care unit in the North West of England. Patients Twenty five children with moderate to severe closed traumatic brain injury and intraparenchymal intracranial pressure monitoring in intensive care (2 -17 years of age). Interventions Routine nursing care interventions. Measurements and main results ICP measured one minute before the procedure, at the maximal value during the procedure and five minutes after the procedure was recorded for the purpose of this study. Time to recovery was also recorded, in minutes. A total of 25 measurements (the first one in each child) in the first 36 hours of the child's PICU admission were analysed. Both ETSMV and log-rolling were associated with clinically and statistically significant changes in ICP from baseline to maximal ICP (p=0.005) and maximal to 5-minute post ICP (p=0.001) for ETSMV and (p < 0.001) baseline to maximal ICP and (p=0.002) for maximal to post-procedure ICP for log-rolling. During ETSMV and logrolling 70% of children exceeded the 20mmHg clinical treatment threshold during the interventions. During both ETSMV and log-rolling children with higher baseline ICPs ( > 15mmHg) showed higher maximal ICPs (but not ICP rise), suggesting a linear relationship between baseline and maximal ICP, although this was more pronounced during turning. One third of the children had not returned to their baseline ICP by 5 minutes after ETSMV, compared with 60% children after log-rolling. Neither eye care nor mouth care showed any clinically significant effects on ICP in these children, suggesting these procedures are not noxious and are tolerated very well. However, there was only a small number of washing episodes reported in this study therefore the observations are not conclusive. Conclusions Endotracheal suctioning and log-rolling in moderate to severe traumatic brain injured children can cause significant intracranial instability and should only be performed as required and with careful planning and execution. Eye and mouth care and washing appear to be well tolerated interventions and could be performed when necessary.

Background: Sedation withdrawal is one of the terms used to describe the behavioural response to stopping or reducing sedative drugs in physically dependent patients. Withdrawal behaviours differ according to the drug involved and may be unpleasant and interfere with recovery. Recognition of sedation withdrawal is challenging due to differences in patient presentation and may be further complicated by the patient’s condition and concomitant drug therapy. Overall Aim of the full thesis: To improve the accuracy of sedation withdrawal assessment in critically ill children. Objectives and Methods: A mixed methods interactive approach comprising six studies. Study 1 evaluates the psychometric properties of the Sedation Withdrawal Score, Studies 2 and 3 examine the complexities/challenges of withdrawal assessment by critiquing existing tool validation studies, A further three studies examine the nurse and parent perspectives of sedation withdrawal assessment in critically ill children. Study 4 investigates how nurses use a sedation withdrawal tool, Studies 5 and 6 investigate what behavioural signs parents recognise and ascertain parents’ willingness to participate in withdrawal assessments. Key findings: Nurses found withdrawal behaviours difficult to interpret in critically ill children and there were differences in how these behaviours were construed. Parents identified a broader range of behaviours than included in existing tools. Most parents were eager to participate in the assessment. The elusive theoretical basis for the existing approach to withdrawal assessment may account for the lack of a standardisation and poor accuracy of the current tools. A model of the causal relationship between dependence and withdrawal is proposed. Recommendations: The model identifies the diagnostic criteria upon which a definition for Pediatric Withdrawal Syndrome may be based. These criteria also provide a novel framework for withdrawal assessment. Focussing on the shared diagnostic criteria and including the parent perspective of the child’s behaviours may aid the assessment and support decision-making.

Made available in DSpace on 2014-06-11T19:29:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2007-06-21Bitstream added on 2014-06-13T18:59:02Z : No. of bitstreams: 1 araujo_db_me_botfmvz.pdf: 249258 bytes, checksum: 539ad5602bdd6df582591ea9174f378e (MD5) / Fundação para o Desenvolvimento da UNESP (FUNDUNESP) / A utilização de métodos sensíveis e específicos para o diagnóstico da raiva constitui uma importante ferramenta para o controle e profilaxia dessa enfermidade. A Reação em Cadeia pela Polimerase através de Transcriptase-Reversa (RT-PCR) tem sido utilizada com bons resultados no diagnóstico do vírus rábico, mesmo quando as amostras estão em estágio de decomposição. Adicionalmente as técnicas moleculares têm sido utilizadas para estudos epidemiológicos possibilitando um melhor conhecimento da epidemiologia viral. O presente trabalho teve como objetivo avaliar as técnicas de RT-PCR e hnRT-PCR para a detecção do vírus rábico em estudos retrospectivos, para isso foram avaliadas 101 amostras cerebrais de diferentes espécies animais armazenadas por diferentes períodos, recém-descongeladas e mantidas em temperatura ambiente por 72 horas para decomposição. Os resultados das técnicas de RT-PCR e hnRT-PCR foram comparados com resultados prévios da Imunofluorescência Direta (IFD) e Inoculação Intracerebral em Camundongos (IC). Das 50 amostras positivas testadas, 26 (52%) apresentaram resultados positivos para a RT-PCR e 45 (90%) com a associação da hnRT-PCR quando realizadas em amostras recentemente descongeladas. Das amostras em decomposição foram analisadas 48 previamente positivas; onde 17 (34,3%) apresentaram resultado positivo para a RT-PCR e 36 (75%) com a associação da hnRT-PCR. Não foram encontrados resultados falso-positivos nas amostras negativas submetidas às técnicas de biologia molecular. A hnRT-PCR apresentou maior sensibilidade em relação à RT-PCR nas amostras recém-descongeladas e em decomposição. Os resultados sugerem a viabilidade de sua aplicação em estudos retrospectivos em materiais descongelados e decompostos. / The use of methods, both sensitive and specific, for rabies diagnosis are important tools for the control and prophylaxis of the disease. Reverse-Transcriptase Polymerse Chain Reaction (RT-PCR) has been used in rabies diagnosis with good results, even in decomposed materials. Additionally, molecular techniques have been used for epidemiological studies and better knowledge of viral epidemiology. The aim of this work was to evaluate the RT-PCR and hnRT-PCR in rabies virus detection in retrospectives studies. RT-PCR and hnRT-PCR were evaluated in 151 brain samples from different animal species, thawed and left at room temperature for 72 hours for decomposition. The RT-PCR and hnRT-PCR results were compared with preview results from Fluorescent Antibody Test and Mouse Inoculation Test. From the 50 positive fresh samples, 26 (52%) were positive for RT-PCR and 45 (90%) for hnRT-PCR. From the 48 positive decomposed samples, 17 (34, 3%) were positive for RT-PCR and 36 (75%) for hnRT-PCR. No false-positives results were found in the negatives samples submitted to the molecular techniques. These results show that the hnRT-PCR was more sensitive than RT-PCR, and both techniques presented lower sensibility in decomposed samples. The hnRT-PCR presented greater sensibility than the standards techniques (IFD e IC) in decomposed materials for rabies diagnosis. These results suggest the viability of the application of molecular techniques in thawed and decomposed materials for retrospective studies.

Dentre as oito espécies do gênero Listeria, a única transmitida por alimentos, patogênica para humanos, é a Listeria monocytogenes. De grande preocupação para a indústria alimentícia, este microrganismo pode ser encontrado em diversas superfícies de plantas processadoras de alimentos, sendo capaz de se aderir e formar persistentes biofilmes. No presente trabalho, foi estudada a influência da concentração de glicose e do tempo de incubação na formação de biofilme por uma cepa de L. monocytogenes, sorotipo 1/2a, cultivada em três diferentes temperaturas, 7°C, 25°C e 37°C, através de planejamento experimental utilizando a Metodologia de Superfície de Resposta. As duas variáveis testadas, para os intervalos estudados, foram significativas (p<0,05) na formação de biofilme somente na temperatura de 37°C e, tanto concentrações de glicose tendendo a zero combinadas com tempos de incubação próximos a 26 h, como maiores concentrações de glicose combinadas com menores tempos de incubação, dentro da faixa de estudo aplicada, representaram boas condições para a formação de biofilme. Foi também realizada a análise transcricional de genes relacionados à formação de biofilme e virulência em duas cepas de L. monocytogenes, sorotipos 1/2a e 4b, cultivadas a 7°C e 37°C (condição controle), pela técnica de PCR quantitativo em tempo real (RT-qPCR). Para ambas as cepas, os genes sigB, prfA, luxS, sufS, sufU, ltrC e flaA apresentaram transcrição significativamente aumentada (p<0,05) a 7°C, em comparação aos resultados para a condição de cultivo a 37°C, enquanto que o gene hly não variou significativamente a sua transcrição entre as duas temperaturas. O gene actA foi mais transcrito a 7°C para a cepa denominada 55, do sorotipo 1/2a, enquanto que não variou significativamente a sua transcrição entre as duas condições de crescimento para a cepa denominada 47, do sorotipo 4b. No entanto, para a cepa 55, o gene degU não demonstrou diferença estatisticamente significativa entre seus níveis de transcrição nas duas temperaturas estudadas, mas para a cepa 47, apresentou transcrição significativamente aumentada a 7°C. Interessantemente, a presença do gene agrA não foi detectada na cepa 47, e os seus níveis de transcrição foram menores a 7°C para a cepa 55. Estes resultados demonstram que os dois sorotipos estudados, responsáveis por muitos dos casos humanos de listeriose, apresentam mecanismos moleculares diferentes, nas temperaturas de 7°C e 37°C. / Among the eight species of genus Listeria, the only transmitted by food, pathogenic for humans, is Listeria monocytogenes. Of great concern to the food industry, this microorganism can be found in various areas of food processing plants, being able to adhere and form persistent biofilms. In the present work, the influence of glucose concentration and incubation time on biofilm formation by a strain of L. monocytogenes, serotype 1/2a, grown in three different temperatures, 7°C, 25°C and 37°C, have been studied, through an experimental design using the Response Surface Methodology. The two variables tested, for the studied intervals, were significant (p<0,05) in the biofilm formation only at a temperature of 37°C and, both glucose concentrations tending to zero combined with incubation times near to 26 h, and higher glucose concentrations combined with lower incubation times, within the applied range study, represented good conditions for biofilm formation. Transcriptional analysis of genes related to biofilm formation and virulence in two strains of L. monocytogenes, serotypes 1/2a and 4b, grown at 7°C and 37°C (control condition), was also performed, by real-time quantitative PCR (RT-qPCR). For both strains, sigB, prfA, luxS, sufS, sufU, ltrC and flaA genes showed significantly increased transcription (p<0,05) at 7°C, in comparison with results for the growth condition at 37°C, whereas hly gene did not varied significantly its transcription between the two temperatures. The actA gene was more transcribed at 7°C for the 55 strain, serotype 1/2a, while did not varied significantly its transcription between the two growth conditions for the 47 strain, serotype 4b. However, for 55 strain, the degU gene did not showed statistically significant difference for its transcription levels between the two studied temperatures, but for 47 strain, presented significantly increased transcription at 7°C. Interestingly, the presence of agrA gene was not detected in 47 strain, and its transcription levels were lower at 7°C for 55 strain. These results demonstrate that the two studied serotypes, responsible for many of the human listeriosis cases, have different molecular mechanisms, at temperatures of 7 and 37°C.