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Cloninger’s Psychobiological Theory of Personality proposes four temperament dimensions, each underpinned by a different neurotransmitter system. The serotonergic system is purportedly linked to Harm Avoidance (HA). The aim of this study was to explore the relationship between HA and serotonin in migraine without aura (MO). A second aim was to explore the personality profile of MO patients. Sixty-six participants completed an online questionnaire and donated blood samples. Results indicated no significant association between HA and serotonin and a significant relationship between MO and HA. This study indicates that both Cloninger’s Psychobiological Theory of Personality and the Tridimensional Personality Questionnaire used for its assessment have value in South African personality research. In addition, the findings of the study reveal support for personality influences on the processes involved in migraine. This not only produces worthwhile avenues of research but also an alternative perspective for clinical practice. / Psychology / M.A. (Psychology (Research Consultation))

O objetivo deste estudo foi avaliar a contribuição de linfoproliferação (1 parâmetro) em associação a provas sorológicas de alto desempenho (4 parâmetros), estudo epidemiológico (1) e parasitológico (1) em indivíduos com sorologia convencional inconclusiva para a doença de Chagas. Mostramos que o diagnóstico de doença de Chagas é provável quando 3 ou mais desses parâmetros são positivos em 7 (15/73). A combinação: TESA-blot e linfoproliferação positivos revelou-se útil diante de antecedentes epidemiológicos positivos / This study aims to evaluate the contribution of lymphoproliferation (1 parameter) in association with high performance serological tests (4), epidemiological data (1) and parasitological tests (1) for Chagas disease in patients with inconclusive conventional serological tests. We showed that this diagnosis is probable in individuals presenting > three positive of these 7 parameters (15 of 73 individuals). The combination of TESA-blot, lymphoproliferation was useful when epidemiological data were positive

Neste estudo avaliamos a resposta imune humoral de indivíduos naturalmente expostos à malária em áreas endêmicas no Brasil. Os anticorpos IgG, IgG1, IgG2, IgG3, IgG4, IgM, IgE e IgA anti-formas eritrocitárias de Plasmodium falciparum foram determinadas por ELISA. Anticorpos IgG, IgG1, IgG2 de alta avidez e IgG3 de baixa avidez predominaram nos indivíduos sem complicações de malária ou assintomáticos, enquanto anticorpos IgG4, IgE e IgM predominaram nos indivíduos com complicações clínicas por malária. Os resultados mostram que mesmo em regiões com transmissão instável de malária pode ser observado o desenvolvimento de imunidade protetora quando anticorpos apropriados são produzidos / In this study, we have evaluated the humoral immune response of individuals naturally exposed to malaria living in endemic areas of Brazil. We determined IgG, IgG1, IgG2, IgG3, IgG4, IgM, IgE and IgA antibodies against Plasmodium falciparum blood stages by ELISA. We observed that the level of high avidity IgG, IgG1 and IgG2 and low avidity IgG3 antibodies were higher in asymptomatic individuals or with uncomplicated malaria, while IgG4, IgE and IgM antibodies were higher in individuals with complicated malaria. Taken together the results showed that even in unstable malaria regions it can be observed the development of protective immunity against malaria when appropriate antibodies are produced

Abstract Endometrial carcinoma is the most common gynegologic malignancy in developed countries. Due to early symptoms, including abnormal uterine bleeding, endometrial cancer is often diagnosed at an early stage and in that case usually has a good prognosis and high cure rates. However, the nature of the disease is heterogeneous. During the last decades, the improvement in survival rates among endometrial cancer patients has not been significant, suggesting that the traditional clinicopathological factors may be inadequate to identify patients with high-risk disease. Furthermore, aggressive adjuvant treatments can be costly and very toxic. Therefore, better prognostic markers associated with biological aggressiveness of endometrial carcinoma are needed to identify the patients with high-risk disease, and to be able to select the treatment more individually. Gelatinases (MMP-2 and MMP-9) and their tissue inhibitors (TIMP-1 and TIMP-2) have been found to play a role in tumor progression. In the present work, the expression and prognostic value of MMP-2, MMP-9, TIMP-1 and TIMP-2 were assessed in endometrial carcinoma. The patient material consisted of a total of 266 women diagnosed with primary endometrial carcinoma. The tissue expression of immunoreactive proteins was examined in paraffin-embedded tumor sections by immunohistochemical staining using specific antibodies, and the pretreatment serum levels of the proteins were quantitatively measured by ELISA. Tissue MMP-2 expression associated with a worsened prognosis, whereas tissue TIMP-2 overexpression was an indicator of a favorable outcome. Furthermore, we observed a combination of strong MMP-2 and weak TIMP-2 tissue expression to identify a group of women at high risk of adverse outcome in endometrial carcinoma. Patients with negative MMP-2 immunostaining had the best prognosis, regardless of TIMP-2 staining result. In serum measurements, high preoperative TIMP-1 concentration was a prognostic indicator of unfavorable outcome. These results indicate that tissue MMP-2 and TIMP-2 as well as circulating TIMP-1 may be prognostic markers in endometrial carcinoma. Of these, tissue MMP-2 seems to be the most potent prognostic marker. Studies with larger patient materials are needed to further explore the value of these enzymes in clinical practice in endometrial cancer. / Tiivistelmä Kohdunrungon syöpä on yleisin gynekologinen maligniteetti kehittyneissä maissa. Varhaisten oireiden, kuten poikkeavan verisen vuodon, vuoksi kohdunrungon syöpä havaitaan usein varhaisessa vaiheessa, jolloin sen ennuste on hyvä. Taudin käyttäytyminen voi kuitenkin olla moninaista. Viime vuosikymmenten aikana kohdunrungon syöpään sairastuneiden ennuste ei ole merkittävästi parantunut. Vaikuttaisi siltä, että perinteiset ennustetekijät eivät ole riittävän tarkkoja ennustamaan syövän taudinkulkua. Lisäksi liitännäishoidot voivat olla kalliita, ja niihin voi liittyä vakavia haittavaikutuksia. Uusien biologisten ennustetekijöiden löytäminen olisi tärkeää, jotta aggressiivista syöpätyyppiä sairastavat potilaat pystyttäisiin tunnistamaan entistä paremmin, ja hoito kyettäisiin räätälöimään yksilöllisemmin taudinkuvaa vastaavasti. Gelatinaasien (MMP-2 ja MMP-9) sekä niiden kudosinhibiittoreiden (TIMP-1 ja TIMP-2) on havaittu osallistuvan syövän etenemiseen. Tässä tutkimuksessa tarkasteltiin MMP-2:n ja MMP-9:n sekä niiden kudosinhibiittoreiden TIMP-1:n ja TIMP-2:n ilmentymistä ja ennusteellista merkitystä kohdunrungon syövässä. Aineisto käsitti yhteensä 266 primaariseen kohdunrungon syöpään sairastunutta naista. Määritysmenetelminä käytettiin sekä immunohistokemiallista värjäystä parafiiniin valettujen kudosnäytteiden osalta että ELISA-määrityksiä ennen hoitoa otettujen seeruminäytteiden osalta. Syöpäkudoksen runsas MMP-2 -proteiinin ilmentyminen liittyi epäsuotuisaan ennusteeseen, kun taas kasvainkudoksen voimakas TIMP-2 -proteiinin ilmentyminen oli hyvän ennusteen merkki. Lisäksi kasvainkudoksen voimakkaan MMP-2- ja heikon TIMP-2 -proteiinien ilmentymisen yhdistelmän havaittiin liittyvän suurempaan syövästä johtuvaan kuolleisuuteen. MMP-2 -negatiivisten potilaiden eloonjäämisennuste oli paras, TIMP-2 -värjäystuloksesta riippumatta. Seerumin korkea TIMP-1 -pitoisuus oli merkittävä huonontuneen ennusteen merkki. Tutkimuksen tulokset viittaavat siihen, että kasvainkudoksessa esiintyvät MMP-2- ja TIMP-2 -proteiinit samoin kuin seerumin TIMP-1 -pitoisuus voivat ennustaa kohdunrungon syövän kliinistä käyttäytymistä. Kasvainkudoksessa esiintyvä MMP-2 -proteiini vaikuttaisi olevan merkittävin ennusteellinen tekijä, mutta tulosten varmistamiseksi tarvitaan lisää tutkimuksia suuremmilla potilasaineistoilla.

Along with proteins, a myriad of different molecular biomarkers, such as post-translational modifications and autoantibodies, could be used in an attempt to improve disease detection and progression. In this thesis, I build on several iterations of the proximity ligation assay to develop and apply new adaptable methods to facilitate detection of proteins, autoantibodies and post-translational modifications. In paper I, we present an adaptation of the solid-phase proximity ligation assay (SP-PLA) for the detection of post-translational modification of proteins (PTMs). The assay was adapted for the detection of two of the most commons PTMs present in proteins, glycosylation and phosphorylation, offering the encouraging prospect of using detection of PTMs in a diagnostic or prognostic capacity.  In paper II, we developed a variant of the proximity ligation assay using micro titer plate for detection and quantification of protein using optical density as readout in the fluorometer, termed PLARCA. With a detection limit considerably lower than ELISA, PLARCA detected femtomolar levels of these proteins in patient samples. In paper III, we aim to compare detection values of samples collected from earlobe capillary, venous plasma, as well as capillary plasma stored in dried plasma spots (DPS) assessed with a 92-plex inflammation panel using multiplex proximity extension assay (PEA). Despite the high variability in protein measurements between the three sample sources, we were able to conclude that earlobe capillary sampling is a suitable less invasive alternative, to venipuncture. In paper IV, we describe the application of PLARCA and proximity extension assay (PEA) for the detection of GAD65 autoantibodies (GADA). Thus, offering highly sensitive and specific autoimmunity detection.

Cardiovascular diseases are a broad group of diseases, such as heart attack and heart failureaffecting the cardiovascular system. The primary cause of cardiovascular diseases isatherosclerosis, and its progression is brought about by oxidative stress and a complex chronicinflammation reaction cascade. Of central importance are proinflammatory cytokines, regulatedby multiple factors, including interleukin (IL) 17A. This project aims to investigate the effectof IL-17A on the inflammatory response of human vascular endothelial cells by quantifyingchemokine C-X-C motif ligand-1 (CXCL1) release when exposed or not to otherproinflammatory mediators such as TNF-𝛼, IL-6 and IL-1β. To investigate this, humanumbilical cord endothelial cells were cultured and then stimulated with IL-17A alone or incombination with other cytokines, namely IL-6/sIL6R, IL-1β, or TNF-𝛼. After an appropriateincubation time following the stimulations, the supernatants of the cells were collected, and theamount of CXCL1 was analysed with ELISA or qPCR, respectively. At a lower concentration(10ng/ml), IL-17A failed to induce a significant level of CXCL1 release from endothelial cells.However, IL-17A + TNF-𝛼 (5ng/ml) greatly enhanced, higher than inductions from individualtreatments combined, level of CXCL1 release from endothelial cells. Furthermore, combiningIL-17A with IL-1β or IL-6 induced non-abundant and abundant upregulation in CXCL1 release,respectively. On transcription level, the amount of CXCL1 mRNA induced by IL-17A alonewas non-significant, but stimulation with TNF-𝛼 and IL-17A + TNF-𝛼 induced significantlyupregulated expression of CXCL1. In conclusion, we found that IL-17A induced synergeticrelease of CXCL1 in human vascular endothelial cells with TNF-𝛼. In addition, the synergisticimpact of IL-17A and TNF-𝛼 in terms of CXCL1 induction in vascular endothelial cells wasevident on a transcriptional level. Our data imply that combined blockage of IL-17A and TNF-𝛼 could have an enhanced therapeutic effect on vascular inflammation.

BACKGROUND: Numerous studies have suggested that Helicobacter pylori infection in asymptomatic subjects is transmitted from person to person. Its prevalence is higher in the institutionalized setting. If that is the case, persons involved in patient care should have a higher prevalence of the infection. METHODS: We estimated the prevalence of H pylori antibodies among groups of asymptomatic medical and nursing staff and compared them with volunteer blood donors of similar age and sex. RESULTS: One hundred fifty-eight nurses and aides, 59 residents, 46 senior medical students, and 22 senior nursing students were enrolled in this study. Serum samples were tested for IgG antibodies against H pylori by enzyme-linked immunosorbent assay. Sixty-two (39%) of 158 nurses were found to be positive for antibodies to H pylori compared with 114 (26%) of 441 specimens from the blood donor group. Within the youngest age group (20 to 34 years), 13 (25%) of 51 nurses were positive for H pylori antibodies compared with 19 (13%) of 143 age-matched serum samples from the blood donor group. Within the middle age group (35 to 49 years), 32 (39%) of 83 nurses were positive for H pylori antibodies vs 43 (26%) of 167 age-matched blood donors. In the oldest age group (> 50 years), 17 (71%) of 24 nurses were positive for H pylori antibodies compared with 52 (40%) of 131 age-matched blood donors. Twenty-three (27%) of 86 nurses with 1 to 15 years of occupational exposure were positive for H pylori antibodies compared with 40 (56%) of 72 nurses with more than 15 years of occupational exposure. CONCLUSIONS: Nurses have an increased prevalence of H pylori antibodies that is significantly higher than the comparable prevalence of volunteer blood donors and is evident in the youngest age group. In addition, the increased prevalence is related to a longer duration of patient exposure in the nursing group.

Introdução: Apesar dos avanços nas técnicas de diagnóstico e tratamento, o câncer de próstata avançado ainda é uma condição letal. Terapêuticas mais eficazes são necessárias para reduzir as taxas de morbi-mortalidade associadas à doença. A Proteína-78 regulada pela glicose (GRP78), uma proteína de choque térmico envolvida na apresentação de antígenos, foi recentemente descrita como sendo um possível marcador molecular para o câncer de próstata. Ainda mais, a resposta imune a essa proteína mostrou correlação com o desenvolvimento de doença hormônio-independente e com pior sobrevida para a doença. Objetivos: Neste estudo, avaliou-se a hipótese de que a GRP78 poderia ser usada como marcador molecular em câncer de próstata no desenvolvimento de um sistema de receptor-ligante, através do uso da tecnologia de apresentação de fagos. Casuística e métodos: Inicialmente, foram clonados dois peptídeos que apresentam afinidade à proteína regulada pela GRP78 (os peptídeos WIFPWIQL e WDLAWMFRLPVG) no vetor fUSE5, criando-se fagos com capacidade teórica de ligação à mesma proteína. Posteriormente foi testada a capacidade de ligação desses fagos à GRP78 na membrana de células prostáticas malignas em solução, em xeno-tumores in vivo e em metástases ósseas de câncer de próstata humano. Resultados: Demonstrou-se que ambos os fagos se ligam especificamente à GRP78 in vitro, em comparação à proteínas com seqüência semelhante (proteínas de choque térmico 70 e 90) e não semelhante (albumina sérica bovina). Em seguida, mostrou-se que esses fagos se ligam com afinidade pelo menos 30 vezes maior à células de câncer de próstata que o fago controle, e que os fagos são internalizados por essas células. Posteriormente, mostrou-se que os fagos rastrearam xeno-tumores prostáticos quando injetados in vivo num modelo animal de câncer de próstata. Finalmente, mostrou-se que os fagos ligam-se especificamente à GRP78 expressa em metástases ósseas de adenocarcinoma prostático humano. Conclusões: Os fagos criados apresentam capacidade de ligação específica à GRP78 in vitro, em células em suspensão e in vivo. A estratégia e o sistema de receptor-ligante definidos no presente estudo podem ter implicacões relevantes no desenvolvimento de terapias dirigidas para o tratamento do câncer de próstata. / Introduction: Despite the advances in diagnosis and treatment, advanced prostate cancer remains a lethal condition. Improved methods of therapy are needed to reduce the morbidity and mortality rates associated with this disease. The Glucose-regulated protein-78 (GRP78), a stress-responsive heat-shock protein involved in antigen presentation, was recently described as a possible molecular marker for prostate cancer. Moreover, immune response against this protein was shown to have correlation with the development of androgen-independent prostate cancer and shorter overall survival. Objectives: We hipothesized that GRP78 could be used as a molecular marker for prostate cancer in the development of a receptor-ligand system, by using phage display technology. Patients and methods: We initially cloned two GRP78-targeting peptides (WIFPWIQL and WDLAWMFRLPVG) into a fUSE5-based phage. We then tested binding capacity of the phage to GRP78 in vitro, to GRP78 expressed in intact prostate cancer cell membranes, to a prostate cancer xenograft and to human bone metastases. Results: We showed that both phage created bound specifically to GRP78 in vitro, in comparison to related (Heat-shock proteins 70 and 90) and unrelated control proteins (bovine serum albumin). Next, we showed that these phage bound at least 30 times more to prostate cancer cells than the control phage, and were also internalized into these cells. Both GRP78-binding phage showed a strong homing in vivo to a human prostate cancer xenograft in a mouse model. Finally, we showed that both phage bound specifically to GRP78 expressed in human prostate cancer bone metastases. Conclusions: Both phage are capable of binding specifically to GRP78 in vitro, in the context of intact prostate cancer cells and in vivo. The strategy and the ligand-receptor system we have defined in this study may have relevant implications in the development of targeted therapies for the treatment of prostate cancer.

Com o objetivo de avaliar o desempenho do teste rápido DPP® LVC comparando com os testes de ELISA e RIFI (Bio-Manguinhos, Br), assim como ELISA e RIFI in-house, empregando como antígeno formas promastigotas de L. (L.) infantum chagasi, com ênfase a reatividade cruzada com outros agentes infecciosos, soros de cães infectados por L. (L.) infantum chagasi, clinicamente sintomáticos (n=48) e assintomáticos (n=39), assim como soros de cães sadios e não infectados (n=18), e soros de cães infectados por Babesia canis (n=9), Dirofilaria immitis (n=4), Trypanosoma cruzi (n=6), Ehrlichia canis (n=17), Neospora caninum (n=6), Toxoplasma gondii (n=9), Neospora/Toxoplasma coinfecção (n=4) e Toxocara canis (n=9) foram avaliados pelas diferentes técnicas de diagnóstico. DPP e ELISA in-house mostraram alta sensitividade (90.81% e 94.25%) e especificidade (95.06% e 97.53%), respectivamente para o diagnóstico de LVC sintomática e assintomática, mas apresentaram reação cruzada com Babesia canis, 44% para DPP e 22% para ELISA in-house. Os dois testes mostraram uma excelente concordância de resultados (kappa=0.9405, p < 0.0001). ELISA Bio-Manguinhos assim como o RIFI Bio-Manguinhos e RIFI in-house mostraram boa sensitividade (90.81%, 96.47% e 89.41%), mas baixa especificidade (77.78%, 69.14% e 65.82%), respectivamente; e mostraram reação cruzada com soros de animais infectados com Babesia canis, Dirofilaria immitis, Trypanosoma cruzi, Ehrlichia canis, Neospora caninum, Toxoplasma gondii. Os resultados mostraram que o DPP® CVL apresentou um bom desempenho para o diagnóstico da leishmaniose visceral canina sintomática e assintomática / In order to investigate the performance of the DPP® CVL rapid test comparing with ELISA and IFA (Bio-Manguinhos, Br), as well as ELISA and IFAT in house using L. (L.) infantum chagasi promastigotes as antigen with emphasis in the cross-reactivity with others infectious agents, sera from clinically symptomatic (n=48) and asymptomatic (n=39) L. (L.) infantum chagasi infected dogs, as well as from healthy non-infected (n=18) dogs and from Babesia canis (n=9), Dirofilaria immitis (n=4), Trypanosoma cruzi (n=6), Ehrlichia canis (n=17), Neospora caninum (n=6), Toxoplasma gondii (n=9), Neospora/Toxoplasma co-infection (n=4) and Toxocara canis (n=9) infected dogs were tested for different diagnosis techniques. DPP and ELISA in-house showed high sensitivity (90.81% and 94.25%) and specificity (95.06% and 97.53%), respectively for symptomatic and asymptomatic CVL diagnosis, but presented cross-reactivity with Babesia canis, 44% for DPP and 22% for ELISA in-house. Both test showed an excellent agreement (kappa=0.9405, p < 0.0001). ELISA Bio-Manguinhos as well as IFA Bio-Manguinhos and IFA in-house showed good sensitivity 90.81%, 96.47% and 89.41%) but low specificity (77.78%, 69.14% and 65.82%), respectively; and showed cross-reactivity with sera from animals infected with Babesia canis, Dirofilaria immitis, Trypanosoma cruzi, Ehrlichia canis, Neospora caninum, Toxoplasma gondii. The results showed that DPP® CVL had a good performance for the diagnosis of of both symptomatic and asymptomatic canine visceral leishmaniasis