Stato delle conoscenze e degli studi sull'etiologia dell'afta epizootica, proprietà del virus, profilassi vaccinale ecc., alla costituzione della commissione : rassegna bibliografica.

At head of title: Ministero dell'interno, Direzione generale della sanità pubblica, Commissione per gli studi e gli esperimenti di immunizzazione contro l'afta epizootica.

Etiologia da doença periodontal

A doença periodontal é caracterizada como um conjunto de condições inflamatórias, de carater crônico ou agudo, e de origem bacteriana, que começa por afetar o tecido gengival e pode levar, com o tempo, à perda dos tecidos de suporte dos dentes. As reações inflamatórias e imunológicas à placa bacteriana representam as características predominantes da gengivite e da periodontite. A reação inflamatória é visível, microscópica e clinicamente, no periodonto afetado e representa a reação do hospedeiro à microbiota da placa e seus produtos. O processo de infecção no sulco periodontal leva, inicialmente, a formação de uma mucosite periodontal, que pode ser definida como uma inflamação dos tecidos moles periodontáis, sem ocasionar perda óssea, sendo reversível, se o seu diagnóstico for atempado. Os processos inflamatórios e imunológicos atuam nos tecidos gengivais para proteger contra o agressãoes microbianas, impedindo os microrganismos de se disseminarem ou invadirem os tecidos. Em alguns casos, essas reações de defesa do hospedeiro podem ser prejudiciais porque também são passíveis de danificar as células e estruturas vizinhas do tecido conjuntivo. Além disso, as reações inflamatórias e imunológicas cuja extensão alcança níveis mais profundos do tecido conjuntivo, além da base do sulco, podem envolver o osso alveolar nesse processo destrutivo. Assim, tais processos defensivos podem, paradoxalmente, ser os responsáveis pela maior parte da lesão tecidual observada na gengivite e na periodontite. O objectivo desse trabalho é fazer uma revisão de literatura específica sobre a etiologia da doença periodontal respectivamente. Serão descritos os principais agentes microbianos que estão relacionados com a doença periodontal e a forma como influenciam o desenvolvimento da doença, procurando desta forma contribuir para a procura de tratamentos mais eficientes.

Uveítes felinas : etiologia e abordagem clínica

Dissertação de Mestrado Integrado em Medicina Veterinária

Etiologia da recessão gengival

Introdução: A Periodontologia é um ramo da Medicina Dentária que tem como objetivo manter o periodonto saudável. A recessão gengival tem vindo a ser estudada, tanto em populações com pobre controlo de placa bacteriana, quanto naquelas com boa Higiene Oral. Os médicos dentistas desconhecem ainda muitos dos aspetos da etiologia da recessão gengival e como tal, este assunto foi objeto de muitas conjeturas, nomeadamente a causa da mesma, sendo ainda mais importante, o controle deste problema. Sendo assim, persiste a confusão levantada por várias opiniões e pontos de vista contraditórios, sendo alvo desta dissertação. Objetivos: Esta dissertação tem como objetivo analisar, e verificar a Etiologia da Recessão Gengival, mais precisamente, a sua origem e os fatores que predispõem a mesma. Tendo sido assim, realizada uma revisão bibliográfica, de modo a verificar: quais as causas desta patologia e as suas limitações. Materiais e Métodos: Para a obtenção da informação necessária na realização da presente dissertação, foi efetuada uma pesquisa bibliográfica nas bases de dados da Pubmed, Scielo, o livro Tratado de Peridontia Clinica e Implantologia Oral, o Jornal da Associação Dentária Americana, o livro Peridontia Clinica, o livro de Histologia Básica e o livro Anatomia, Embriologia e Histologia Oral. Para tal, foi realizada a investigação através das seguintes palavras-chaves: “Etiology”, Gingival Recession”, “Periodontitis”, “Dental plaque”, e “Prevalence”. Conclusão: No trabalho realizado, é possível concluir que a Etiologia da recessão é multifatorial e raramente leva à perda do elemento dentário, embora, cause muitos danos por provocar a sensibilidade dentária, devido a perda e retração da gengiva. Esta etiologia leva a uma maior incidência de cáries radiculares, sacrificando o aspecto estético do paciente, e consequentemente, leva a um desconforto psicológico. Subsequentemente a recessão gengival e as suas múltiplas causas, existem atualmente métodos e técnicas, que nos permitem a resolução de alguns dos danos provocados por esta, com o objetivo de criar uma maior probabilidade de eliminação dos fatores causais da mesma.

Zebrafish as a model of Parkinson's disease

Parkinson’s disease (PD) is the second most common neurodegenerative disease among the elderly. Its etiology is unknown and no disease-modifying drugs are available. Thus, more information concerning its pathogenesis is needed. Among other genes, mutated PTEN-induced kinase 1 (PINK1) has been linked to early-onset and sporadic PD, but its mode of action is poorly understood. Most animal models of PD are based on the use of the neurotoxin 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP). MPTP is metabolized to MPP+ by monoamine oxidase B (MAO B) and causes cell death of dopaminergic neurons in the substantia nigra in mammals. Zebrafish has been a widely used model organism in developmental biology, but is now emerging as a model for human diseases due to its ideal combination of properties. Zebrafish are inexpensive and easy to maintain, develop rapidly, breed in large quantities producing transparent embryos, and are readily manipulated by various methods, particularly genetic ones. In addition, zebrafish are vertebrate animals and results derived from zebrafish may be more applicable to mammals than results from invertebrate genetic models such as Drosophila melanogaster and Caenorhabditis elegans. However, the similarity cannot be taken for granted. The aim of this study was to establish and test a PD model using larval zebrafish. The developing monoaminergic neuronal systems of larval zebrafish were investigated. We identified and classified 17 catecholaminergic and 9 serotonergic neuron populations in the zebrafish brain. A 3-dimensional atlas was created to facilitate future research. Only one gene encoding MAO was found in the zebrafish genome. Zebrafish MAO showed MAO A-type substrate specificity, but non-A-non-B inhibitor specificity. Distribution of MAO in larval and adult zebrafish brains was both diffuse and distinctly cellular. Inhibition of MAO during larval development led to markedly elevated 5-hydroxytryptamine (serotonin, 5-HT) levels, which decreased the locomotion of the fish. MPTP exposure caused a transient loss of cells in specific aminergic cell populations and decreased locomotion. MPTP-induced changes could be rescued by the MAO B inhibitor deprenyl, suggesting a role for MAO in MPTP toxicity. MPP+ affected only one catecholaminergic cell population; thus, the action of MPP+ was more selective than that of MPTP. The zebrafish PINK1 gene was cloned in zebrafish, and morpholino oligonucleotides were used to suppress its expression in larval zebrafish. The functional domains and expression pattern of zebrafish PINK1 resembled those of other vertebrates, suggesting that zebrafish is a feasible model for studying PINK1. Translation inhibition resulted in cell loss of the same catecholaminergic cell populations as MPTP and MPP+. Inactivation of PINK1 sensitized larval zebrafish to subefficacious doses of MPTP, causing a decrease in locomotion and cell loss in one dopaminergic cell population. Zebrafish appears to be a feasible model for studying PD, since its aminergic systems, mode of action of MPTP, and functions of PINK1 resemble those of mammalians. However, the functions of zebrafish MAO differ from the two forms of MAO found in mammals. Future studies using zebrafish PD models should utilize the advantages specific to zebrafish, such as the ability to execute large-scale genetic or drug screens.

Socioeconomic Status, Occupational Factors and Lifestyle as Predictors of Hospitalisation for Back Disorders

The aim of this dissertation was to examine the determinants of severe back disorders leading to hospital admission in Finland. First, back-related hospitalisations were considered from the perspective of socioeconomic status, occupation, and industry. Secondly, the significance of psychosocial factors at work, sleep disturbances, and such lifestyle factors as smoking and overweight was studied as predictors of hospitalisation due to back disorders. Two sets of data were used: 1) the population-based data comprised all occupationally active Finns aged 25-64, and included hospitalisations due to back disorders in 1996 and 2) a cohort of employees followed up from 1973 to 2000 having been hospitalised due to back disorders. The results of the population-based study showed that people in physically strenuous industries and occupations, such as agriculture and manufacturing, were at an increased risk of being hospitalised for back disorders. The lowest hospitalisation rates were found in sedentary occupations. Occupational class and the level of formal education were independently associated with hospitalisation for back disorders. This stratification was fairly consistent across age-groups and genders. Men had a slightly higher risk of becoming hospitalised compared with women, and the risk increased with age among both genders. The results of the prospective cohort study showed that psychosocial factors at work such as low job control and low supervisor support predicted subsequent hospitalisation for back disorders even when adjustments were made for occupational class and physical workload history. However, psychosocial factors did not predict hospital admissions due to intervertebral disc disorders; only admissions due to other back disorders. Smoking and overweight predicted, instead, only hospitalisation for intervertebral disc disorders. These results suggest that the etiological factors of disc disorders and other back disorders differ from each other. The study concerning the association of sleep disturbances and other distress symptoms with hospitalisation for back disorders revealed that sleep disturbances predicted subsequent hospitalisation for all back disorders after adjustment for chronic back disorders and recurrent back symptoms at baseline, as well as for work-related load and lifestyle factors. Other distress symptoms were not predictive of hospitalisation.

Ischemic Stroke in Young Adults

Stroke is the second leading cause of death and the leading cause of disability worldwide. Of all strokes, up to 80% to 85% are ischemic, and of these, less than 10% occur in young individuals. Stroke in young adults—most often defined as stroke occurring under the age of 45 or 50—can be particularly devastating due to long expected life-span ahead and marked socio-economic consequences. Current basic knowledge on ischemic stroke in this age group originates mostly from rather small and imprecise patient series. Regarding emergency treatment, systematic data on use of intravenous thrombolysis are absent. For this Thesis project, we collected detailed clinical and radiological data on all consecutive patients aged 15 to 49 with first-ever ischemic stroke between 1994 and 2007 treated at the Helsinki University Central Hospital. The aims of the study were to define demographic characteristics, risk factors, imaging features, etiology, and long-term mortality and its predictors in this patient population. We additionally sought to investigate, whether intravenous thrombolysis is safe and beneficial for the treatment of acute ischemic stroke in the young. Of our 1008 patients, most were males (ratio 1.7:1), who clearly outnumbered females after the age of 44, but females were preponderant among those aged <30. Occurrence increased exponentially. The most frequent risk factors were dyslipidemia (60%), smoking (44%), and hypertension (39%). Risk factors accumulated in males and along aging. Cardioembolism (20%) and cervicocerebral artery dissection (15%) were the most frequent etiologic subgroups, followed by small-vessel disease (14%), and large-artery atherosclerosis (8%). A total of 33% had undetermined etiology. Left hemisphere strokes were more common in general. Posterior circulation infarcts were more common among those aged <45. Multiple brain infarcts were present in 23% of our patients, 13% had silent infarcts, and 5% had leukoaraiosis. Of those with silent brain infarcts, majority (54%) had only a single lesion, and most of the silent strokes were located in basal ganglia (39%) and subcortical regions (21%). In a logistic regression analysis, type 1 diabetes mellitus in particular predicted the presence of both silent brain infarcts (odds ratio 5.78, 95% confidence interval 2.37-14.10) and leukoaraiosis (9.75; 3.39-28.04). We identified 48 young patients with hemispheric ischemic stroke treated with intravenous tissue plasminogen activator, alteplase. For comparisons, we searched 96 untreated control patients matched by age, gender, and admission stroke severity, as well as 96 alteplase-treated older controls aged 50 to 79 matched by gender and stroke severity. Alteplase-treated young patients recovered more often completely (27% versus 10%, P=0.010) or had only mild residual symptoms (40% versus 22%, P=0.025) compared to age-matched controls. None of the alteplase-treated young patients had symptomatic intracerebral hemorrhage or died within 3-month follow-up. Overall long-term mortality was low in our patient population. Cumulative mortality risks were 2.7% (95% confidence interval 1.5-3.9%) at 1 month, 4.7% (3.1-6.3%) at 1 year, and 10.7% (9.9-11.5%) at 5 years. Among the 30-day survivors who died during the 5-year follow-up, more than half died due to vascular causes. Malignancy, heart failure, heavy drinking, preceding infection, type 1 diabetes, increasing age, and large-artery atherosclerosis causing the index stroke independently predicted 5-year mortality when adjusted for age, gender, relevant risk factors, stroke severity, and etiologic subtype. In sum, young adults with ischemic stroke have distinct demographic patterns and they frequently harbor traditional vascular risk factors. Etiology in the young is extremely diverse, but in as many as one-third the exact cause remains unknown. Silent brain infarcts and leukoaraiosis are not uncommon brain imaging findings in these patients and should not be overlooked due to their potential prognostic relevance. Outcomes in young adults with hemispheric ischemic stroke can safely be improved with intravenous thrombolysis. Furthermore, despite their overall low risk of death after ischemic stroke, several easily recognizable factors—of which most are modifiable—predict higher mortality in the long term in young adults.

Peripheral facial palsy : Grading, Etiology, and Melkersson-Rosenthal Syndrome

The objective of this study was to assess the utility of two subjective facial grading systems, to evaluate the etiologic role of human herpesviruses in peripheral facial palsy (FP), and to explore characteristics of Melkersson-Rosenthal syndrome (MRS). Intrarater repeatability and interrater agreement were assessed for Sunnybrook (SFGS) and House-Brackmann facial grading systems (H-B FGS). Eight video-recorded FP patients were graded in two sittings by 26 doctors. Repeatability for SFGS was from good to excellent and agreement between doctors from moderate to excellent by intraclass correlation coefficient and coefficient of repeatability. For H-B FGS, repeatability was from fair to good and agreement from poor to fair by agreement percentage and kappa coefficients. Because SFGS was at least as good in repeatability as H-B FGS and showed more reliable results in agreement between doctors, we encourage the use of SFGS over H-B FGS. Etiologic role of human herpesviruses in peripheral FP was studied by searching DNA of herpes simplex virus (HSV) -1 and -2, varicella-zoster virus (VZV), human herpesvirus (HHV) -6A, -6B, and -7, Epstein-Barr virus (EBV), and cytomegalovirus (CMV) by PCR/microarray methods in cerebrospinal fluid (CSF) of 33 peripheral FP patients and 36 controls. Three patients and five controls had HHV-6 or -7 DNA in CSF. No DNA of HSV-1 or -2, VZV, EBV, or CMV was found. Detecting HHV-7 and dual HHV-6A and -6B DNA in CSF of FP patients is intriguing, but does not allow etiologic conclusions as such. These DNA findings in association with FP and the other diseases that they accompanied require further exploration. MRS is classically defined as a triad of recurrent labial or oro-facial edema, recurrent peripheral FP, and plicated tongue. All three signs are present in the minority of patients. Edema-dominated forms are more common in the literature, while MRS with FP has received little attention. The etiology and true incidence of MRS are unknown. Characteristics of MRS were evaluated at the Departments of Otorhinolaryngology and Dermatology focusing on patients with FP. There were 35 MRS patients, 20 with FP and they were mailed a questionnaire (17 answered) and were clinically examined (14 patients). At the Department of Otorhinolaryngology, every MRS patient had FP and half had the triad form of MRS. Two patients, whose tissue biopsies were taken during an acute edema episode, revealed nonnecrotizing granulomatous findings typical for MRS, the other without persisting edema and with symptoms for less than a year. A peripheral blood DNA was searched for gene mutations leading to UNC-93B protein deficiency predisposing to HSV-1 infections; no gene mutations were found. Edema in most MRS FP patients did not dominate the clinical picture, and no progression of the disease was observed, contrary to existing knowledge. At the Department of Dermatology, two patients had triad MRS and 15 had monosymptomatic granulomatous cheilitis with frequent or persistent edema and typical MRS tissue histology. The clinical picture of MRS varied according to the department where the patient was treated. More studies from otorhinolaryngology departments and on patients with FP would clarify the actual incidence and clinical picture of the syndrome.

Written Cancer Narratives : An Ethnomedical Study of Cancer Patients' Thoughts, Emotions and Experiences

Since the second half of the 20th century, cancer has become a dominant disease in Western countries, endangering people regardless of age, gender, race or social status. Every year almost eight million people die of cancer worldwide. In Finland every fourth person is expected to fall ill with cancer at some stage of his or her life. During the 20th century, along with rapid changes in the medical system, people s awareness of cancer has increased a great deal. This has also influenced the image of cancer in popular discourse over the past decades. However, from the scientific point of view there is still much that is unclear about the disease. This thesis shows that this is a big problem for ordinary people, as, according to culture-bound illness ideology, people need an explanation about the origin of their illness in order to help them cope. The main aim of this thesis is to examine the process of being ill with cancer from the patient s point of view, in order to analyse attitudes and behaviour towards cancer and its significance and culture-bound images. This narrative-based study concentrates on patients voicings , which are important in understanding the cancer experience and when attempting to make it more open within current cultural and societal settings. The Kun sairastuin syöpään ( when I fell ill with cancer ) writing competition organised by Suomen Syöpäpotilaat ry (the Finnish Cancer Patients Association), Suomen Syöpäyhdistys ry (the Finnish Cancer Union), and Suomalaisen Kirjallisuuden Seuran kansanrunousarkisto (the Finnish Literary Society Folklore Archive) was announced on the 1st of May 1994 and lasted until the 30th of September 1994. As a result, a total of 672 cancer narratives, totalling 6384 pages, were received, filled with experiences relating to cancer. Written cancer narratives form a body of empirical data that is suitable for content or textual analysis. In this thesis, content analysis is adopted in order to become familiar with the texts and to preselect the themes and analytical units for further examination. I use multiple perspectives in order to interpret cancer patients ideas and reasoning. The ethnomedical approach unites popular health beliefs that originated in Finnish folk medicine, as well as connecting alternative medicine, which patients make use of, with biomedicine, the dominant form of medicine today. In addition to this, patients narratives, which are composed of various structural segments, are approached from the folklorist s perspective. In this way they can be seen as short pathographies, reconstructions of self-negotiation and individual decision making during the illness process. Above all, cancer patients writing describe their feelings, thoughts and experiences. Factors that appear insignificant to modern medicine, overwhelmed as it is by medical technologies that concentrate on dysfunctional tissue within diseased bodies. Ethnomedical study of cancer patients writings gives access to the human side of cancer discourse, and combines both medical, and popular, knowledge of cancer. In my view, the natural world and glimpses of tradition are bound together with one general aim within cancer narratives: to tackle the illness and mediate its meanings. Furthermore, the narrative approach reveals that participants write with the hope of offering a different interpretation of the cancer experience, and thus of confronting culturally pre-defined images and ideologies.

Kantasolusiirrännäiset hevosen kroonisen nivelrikon hoidossa

Nivelrikko aiheuttaa suuria taloudellisia tappioita ja vaikuttaa sosiaaliseen kanssakäymiseen ja hyvinvointiin. Hevosilla se on yleisin ontuman aiheuttaja, ja ontumat ovat yleisin syy hevosten poistamiseen harjoitus- ja kilpailukäytöstä. Nivelruston uusiutuminen on hyvin rajallista, ja sen vauriot johtavatkin usein nivelrikkoon. Nivelrikon etiologia ei ole vielä täysin selvillä, mutta sen kehittymiseen vaikuttavat ainakin nivelten kehityshäiriöt, niveliin kohdistuvat vammat, nivelruston alla olevan luun muutokset sekä nivelten löysyys. Nivelrikon kehittyessä ruston ulkonäkö ja koostumus muuttuvat. Rustovaurion seurauksena nivelnesteeseen vapautuu ruston hajoamistuotteita, jotka todennäköisesti saavat aikaan sytokiinien erittymisen. Sytokiineistä IL-1 (interleukiini-1) ja TNF-α (tumor necrosis factor-α) saavat aikaan NO (typpimonoksidi) –tuotannon rustosoluissa. NO vähentää rustosolujen kykyä syntetisoida soluväliaineen proteoglykaaneita. NO aiheuttaa PGE2 (prostaglandiini-E2) erityksen, mikä vähentää rustosolujen jakautumista. Alueilla, joilla on korkeat NO-pitoisuudet, on rustossa runsaasti apoptoottisia soluja. IL-1 aktivoi myös metalloproteinaaseja, jotka kiihdyttävät ruston proteoglykaanien katoa. Vaurioitunut rustokudos korvautuu pääosin tiiviillä sidekudoksella. Tämän vuoksi ruston alla olevan luun kuormitus muuttuu ja nivelruston reunoille muodostuu uudisluuta. Nivelrikon hoidon tavoitteena on hillitä tulehdusreaktiota nivelessä ja nivelkapselissa, jolloin myös nivelruston kato vähenee. Konservatiivisesti tähän tavoitteeseen pyritään akuutissa vaiheessa levon, sekä sitä seuraavan fysioterapian ja kontrolloidun liikunnan, tulehduskipulääkityksen, sekä tulehdusta hillitsevien ja nivelrustoa tukevien lääkkeiden ja ravintolisien avulla. Hevosilta kantasoluja voidaan eristää luuytimestä, rasvakudoksesta, aikuisen verestä, napanuoranverestä syntymän yhteydessä, sekä alkioista. Kantasolut saadaan in vitro kasvatusolosuhteita muokkaamalla erilaistumaan rustoksi. Kantasoluja on kokeiltu ihmisillä ja eläimillä (hiiri, kani, koira, sika, lammas) nivelrikon hoitoon. Tulokset ovat olleet ristiriitaisia. Yksittäisen potilasselostuksen mukaan ihmisen polven nivelrikko-oireet helpottivat kantasolusiirrännäisen jälkeen. Toisessa ihmistutkimuksessa potilaiden oireet eivät lievittyneet. Hiirillä ihmisestä peräisin olevien kantasolujen todettiin hillitsevän reumaattisen nivelrikon aiheuttamaa tulehdusreaktiota. Koiratutkimuksessa potilaiden oireilu helpotti. Näissä tutkimuksissa ei selvitetty kantasolujen vaikutusta ruston rakenteeseen. Kaneilla uudiskudoksen todettiin alkavan muistuttaa ajan kuluessa ruston sijaan enemmän sidekudosta. Sioilla ja lampailla kantasolusiirrännäisten seurauksena muodostunut uudiskudos muistutti koostumukseltaan nivelrustoa. Kahdessa julkaistussa hevostutkimuksessa kantasolujen ei todettu parantavan uudiskudoksen rakennetta, eikä uudiskudos ollut rustoa. Kantasolut nopeuttivat alkuvaiheen paranemista. Helsingin Yliopistollisessa eläinsairaalassa on kantasoluilla hoidettu yksi nivelrikkoa sairastanut hevospotilas. Kantasolusiirrosta ei ollut apua tämän potilaan kohdalla, vaan toivottoman ennusteen vuoksi omistajat päätyivät eläinlääkäreiden suosituksesta eutanasiaan. Potilaasta ei saatu rustonäytteitä tutkittavaksi.

Genomic alterations in Myeloproliferative Neoplasms and Myelodysplastic Syndromes

Myeloproliferative neoplasms (MPN) and myelodysplastic syndromes (MDS) are a heterogeneous group of clonal hematopoietic disorders whose etiology and molecular pathogenesis are poorly understood. During the past decade, enormous developments in microarray technology and bioinformatics methods have made it possible to mine novel molecular alterations in a large number of malignancies, including MPN and MDS, which has facilitated the detection of new prognostic, predictive and therapeutic biomarkers for disease stratification. By applying novel microarray techniques, we profiled copy number alterations and microRNA (miRNA) expression changes in bone marrow aspirate and blood samples. In addition, we set up and validated an miRNA expression test for bone marrow core biopsies in order to utilize the large archive material available in many laboratories. We also tested JAK2 mutation status and compare it with the in vitro growth pattern of hematologic progenitors cells. In the study focusing on 100 MPN cases, we detected a Janus kinase 2 (JAK2) mutation in 71 cases. We observed spontaneous erythroid colony growth in all mutation-positive cases in addition to nine mutation negative cases. Interestingly, seven JAK2V167F negative ET cases showed spontaneous megakaryocyte colony formation, one case of which also harbored a myeloproliferative leukemia virus oncogene (MPL) mutation. We studied copy number alterations in 35 MPN and 37 MDS cases by using oligonucleotide-based array comparative hybridization (array CGH). Only one essential thrombocythemia (ET) case presented copy number alterations in chromosomes 1q and 13q. In contrast, MDS cases were characterized by numerous novel cryptic chromosomal aberrations with the most common copy number losses at 5q21.3q33.1 and 7q22.1q33, while the most common copy number gain was trisomy 8. As for the study of the bone marrow core biopsy samples, we showed that even though these samples were embedded in paraffin and underwent decalcification, they were reliable sources of miRNA and suitable for array expression analysis. Further, when studying the miRNA expression profiles of the 19 MDS cases, we found that, compared to controls, two miRNAs (one human Epstein-Barr virus (miR-BART13) miRNA and one human (has-miR-671-5p) miRNA) were downregulated, whereas two other miRNAs (hsa-miR-720 and hsa-miR-21) were upregulated. However, we could find no correlation between copy number alterations and microRNA expression when integrating these two data. This thesis brings to light new information about genomic changes implicated in the development of MPN and MDS, and also underlines the power of applying genome-wide array screening techniques in neoplasias. Rapid advances in molecular techniques and the integration of different genomic data will enable the discovery of the biological contexts of many complex disorders, including myeloid neoplasias.

Identification of collagenolytic MMP networks as potential biomarkers in progressive chronic periodontitis subjects and MMP-8 null allele model

Chronic periodontitis results from a complex aetiology, including the formation of a subgingival biofilm and the elicitation of the host s immune and inflammatory response. The hallmark of chronic periodontitis is alveolar bone loss and soft periodontal tissue destruction. Evidence supports that periodontitis progresses in dynamic states of exacerbation and remission or quiescence. The major clinical approach to identify disease progression is the tolerance method, based on sequential probing. Collagen degradation is one of the key events in periodontal destructive lesions. Matrix metalloproteinase (MMP)-8 and MMP-13 are the primary collagenolytic MMPs that are associated with the severity of periodontal inflammation and disease, either by a direct breakdown of the collagenised matrix or by the processing of non-matrix bioactive substrates. Despite the numerous host mediators that have been proposed as potential biomarkers for chronic periodontitis, they reflect inflammation rather than the loss of periodontal attachment. The aim of the present study was to determine the key molecular MMP-8 and -13 interactions in gingival crevicular fluid (GCF) and gingival tissue from progressive periodontitis lesions and MMP-8 null allele mouse model. In study (I), GCF and gingival biopsies from active and inactive sites of chronic periodontitis patients, which were determined clinically by the tolerance method, and healthy GCF were analysed for MMP-13 and tissue inhibitor of matrix metalloproteinases (TIMP)-1. Chronic periodontitis was characterised by increased MMP-13 levels and the active sites showed a tendency of decreased TIMP-1 levels associated with increments of MMP-13 and total protein concentration compared to inactive sites. In study (II), we investigated whether MMP-13 activity was associated with TIMP-1, bone collagen breakdown through ICTP levels, as well as the activation rate of MMP-9 in destructive lesions. The active sites demonstrated increased GCF ICTP levels as well as lowered TIMP-1 detection along with elevated MMP-13 activity. MMP-9 activation rate was enhanced by MMP-13 in diseased gingival tissue. In study (III), we analysed the potential association between the levels, molecular forms, isoenzyme distribution and degree of activation of MMP-8, MMP-14, MPO and the inhibitor TIMP-1 in GCF from periodontitis progressive patients at baseline and after periodontal therapy. A positive correlation was found for MPO/MMP-8 and their levels associated with progression episodes and treatment response. Because MMP-8 is activated by hypochlorous acid in vitro, our results suggested an interaction between the MPO oxidative pathway and MMP-8 activation in GCF. Finally, in study (IV), on the basis of the previous finding that MMP-8-deficient mice showed impaired neutrophil responses and severe alveolar bone loss, we aimed to characterise the detection patterns of LIX/CXCL5, SDF-1/CXCL12 and RANKL in P. gingivalis-induced experimental periodontitis and in the MMP-8-/- murine model. The detection of neutrophil-chemoattractant LIX/CXCL5 was restricted to the oral-periodontal interface and its levels were reduced in infected MMP-8 null mice vs. wild type mice, whereas the detection of SDF-1/CXCL12 and RANKL in periodontal tissues increased in experimentally-induced periodontitis, irrespectively from the genotype. Accordingly, MMP-8 might regulate LIX/CXCL5 levels by undetermined mechanisms, and SDF-1/CXCL12 and RANKL might promote the development and/or progression of periodontitis.

Avaliação nutricional do paciente com doença hepática crônica

Além de muito freqüente, a desnutrição associa-se a morbi/mortalidade em pacientes com doenças hepáticas crônicas. A avaliação do estado nutricional em hepatopatas é difícil pela sobrecarga hídrica e pela alteração na síntese protéica, fatores que alteram os parâmetros tradicionalmente usados na avaliação nutricional. Os objetivos são:a)avaliar o estado nutricional, através da AGS, antropometria, do escore de Mendenhall e da combinação de todos os instrumentos, em pacientes com doença hepática crônica; b)correlacionar o estado nutricional com a gravidade de doença hepática crônica; c)determinar a contribuição da dinamometria do aperto de mão para a avaliação do estado nutricional. Foram incluídos 305 pacientes portadores de doenças hepáticas crônicas, com idade de 18-80 anos, atendidos no ambulatório de doenças hepatobiliares do Hospital Universitátio Pedro Ernesto. A gravidade da doença hepática foi avaliada pela classificação de Child-Pugh e escore de Meld. Foram aferidos parâmetros antropométricos (peso, altura, índice de massa corporal, prega cutânea triciptal, circunferência do braço, circunferência muscular do braço), parâmetros bioquímicos (albumina e contagem total de linfócitos), Avaliação Global Subjetiva, escore de Mendenhall e força do aperto de mão pela dinamometria. Os valores da porcentagem de adequação dos parâmetros foram utilizados para a classificação da desnutrição. Consideramos todos os pacientes com porcentagens de adequação abaixo de 90% como desnutridos. Foi criado o escore risco de desnutrição que se caracterizou pela alteração em qualquer um dos parâmetros da avaliação nutricional. Cerca de 53% dos pacientes eram do sexo masculino, 43% portadores de cirrose hepática, 80% com etiologia viral e média de idade de 54 12 anos. Houve relação estatisticamente significativa entre a classificação funcional da doença hepática e a AGS, o escore de Mendenhall e o de risco de desnutrição. A avaliação isolada da antropometria não se correlacionou com a classificação funcional. Segundo a AGS, a prevalência de desnutrição foi de 10% na hepatopatia não cirrótica, 16% na cirrose compensada e 94% na cirrose descompensada. Segundo o escore de Mendenhall, as cifras foram de 31%, 38% e 56%, respectivamente. Segundo o novo escore, as cifras foram de 52%, 60% e 96%, respectivamente. Embora tenha havido uma redução estatisticamente significativa da força muscular com o agravamento do estado nutricional, não foi possível estabelecer um ponto de corte para os valores da dinamometria. A análise do desempenho do percentual de adequação da força muscular como critério diagnóstico de pacientes sob risco de desnutrição revelou provavelmente 56% de falso-positivos e 24% de falso-negativos. A grande variação na prevalência de desnutrição em pacientes com doença hepática depende do instrumento de avaliação nutricional usado e da classificação funcional da doença hepática. Não surpreendentemente, os escores combinados detectaram as maiores taxas de prevalência de desnutrição. Houve associação significativa entre o estado nutricional e a gravidade da doença hepática. O aumento das taxas de prevalência de desnutrição trazido pela dinamometria ocorreu às custas de resultados falso-positivos.

Relação entre a doença de Parkinson e o gene LRRK2: um estudo na população brasileira

A doença de Parkinson (DP) é a segunda doença neurodegenerativa mais frequente depois da Doença de Alzheimer, afetando aproximadamente 1% da população com idade superior a 65 anos. Clinicamente, esta doença caracteriza-se pela presença de tremor em repouso, bradicinesia, rigidez muscular e instabilidade postural, os quais podem ser controlados com a administração do levodopa. As características patológicas da DP incluem a despigmentação da substância nigra devido à perda dos neurônios dopaminérgicos e a presença de inclusões proteicas denominadas corpos de Lewy nos neurônios sobreviventes. As vias moleculares envolvidas com esta patologia ainda são obscuras, porém a DP é uma doença complexa, resultante da interação entre fatores ambientais e causas genéticas. Mutações no gene leucine-rich repeat kinase 2 (LRRK2; OMIM 609007) constituem a forma mais comum de DP. Este gene codifica uma proteína, membro da família de proteínas ROCO, que possui, entre outros domínios, dois domínios funcionais GTPase (ROC) e quinase (MAPKKK). Neste estudo, os principais domínios do gene LRRK2 foram analisados em 204 pacientes brasileiros com DP por meio de sequenciamento dos produtos da PCR. Através da análise de 14 exons correspondentes aos domínios ROC, COR e MAPKKK foram identificadas 31 variantes. As alterações novas, p.C1770R e p.C2139S, possuem um potencial papel na etiologia da DP. Três alterações exônicas (p.R1398R, p.T1410M e p.Y2189C) e nove intrônicas (c.4317+16C>T, c.5317+59A>C, c.5509+20A>C, c.5509+52T>C, c.5509+122A>G, c.5657-46C>T, c.6382-36G>A, c.6382-37C>T e c.6576+44T>C) são potencialmente não patogênicas. Ao todo, dezessete variantes exônicas e intrônicas constituem polimorfismos já relatados na literatura (p.R1398H, p.K1423K, p.R1514Q, p.P1542S, c.4828-31T>C, p.G1624G, p.K1637K, p.M1646T, p.S1647T, c.5015+32A>G, c.5170+23T>A, c.5317+32C>T, p.G1819G, c.5948+48C>T, p.N2081D, p.E2108E e c.6381+30A>G). A frequência total de alterações potencialmente patogênicas ou patogênicas detectadas em nossa amostra foi de 3,4% (incluindo a mutação p.G2019S, anteriormente descrita em 2 artigos publicados por nosso grupo: Pimentel et al., 2008; Abdalla-Carvalho et al., 2010), sendo a frequência de mutações nos casos familiares (11,1%) cerca de seis vezes maior do que a encontrada nos casos isolados da DP (1,8%). Os resultados alcançados neste estudo revelam que mutações no gene LRRK2 desempenham um papel significativo como fator genético para o desenvolvimento da DP em pacientes brasileiros.