Candida dubliniensis: prevalência na cavidade bucal de pacientes diabéticos, com câncer de mama e hansenianos e patogenicidade experimental

O objetivo deste estudo foi avaliar a presença de C. dubliniensis dentre isolados bucais de pacientes com Diabetes mellitus do tipo I (n=39) e II (n=36), hansenianos (n=38) e sob quimioterapia para o câncer de mama (n=30) e de respectivos indivíduos controle pareados quanto à idade, gênero e condições bucais. Um total de 479 isolados previamente identificados por testes fenotípicos (formação de tubo germinativo, produção de hifas/pseudohifas/clamidoconídeos, fermentação e assimilação de carboidratos) e identificadas como C. albicans/C. dubliniensis foram incluídos no estudo. A existência de C. dubliniensis dentre os isolados foi analisada usando protocolo validado de PCR multiplex. Foi também realizado estudo de patogenicidade experimental utilizando camundongos em modelo de infecção sistêmica, objetivando comparar a virulência e cinética de infecção de C. dubliniensis com outras espécies do gênero Candida. Um isolado (0.002%) de C. dubliniensis foi detectado entre os isolados do grupo controle. Esta espécie não foi encontrada dentre os isolados dos outros grupos de pacientes. C. dubliniensis foi menos virulenta para camundongos em relação a C. albicans e C. tropicalis e mais virulenta do que C. krusei. O estudo da cinética de infecção mostrou infecção persistente no rim e no fígado mesmo após 21 dias da inoculação de C. dubliniensis. Conclui-se que a detecção de C. dubliniensis dentre os isolados clínicos foi baixa e observada apenas no grupo controle. C. dubliniensis foi menos virulenta para camundongos que C. albicans e C. tropicalis e causou infecção prolongada no rim e no fígado

Evaluation of caries-associated virulence of biofilms from Candida albicans isolated from saliva of pediatric patients with sickle-cell anemia

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Avaliação audiológica comportamental e eletrofisiológica no transtorno do espectro do autismo

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Energy requirements for maintenance and growth of Caninde goat kids

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Dispositivos não convencionais para o rastreamento do corpo humano

Due to the popularity and need to learn sign language, there are tools that help in this type of communication. In Brazil, for example, the government offers professional Sign Language to assist communities, however, if think the hypothesis that people live in places difficult access, there is a difficult in movement of professionals to such places, besides bringing government overspending. Thus, a tool that simulates the movements of professional, fully exploiting the technological resources (cost-benefit) to enrich and provide a tool for communities, it becomes possible order to fill this gap. The system provides resources for editing movements, and these, later, are retrieved by tool for the visualization and representation of signals in a Virtual Environment. The resources used, provide two unconventional devices for tracking body, the Wiimote controllers and P5 glove.

Variabilidade química infraespecífica e dinâmica de metabólitos secundários fenólicos e diterpênicos em Casearia sylvestris Sw. (Salicaceae): correlação metabolômica e proteômica utilizando cromatografia, espectrometria de massas, ressonância magnética nuclear e quimiometria

Pós-graduação em Química - IQ

Documentação química de espécies do gênero Casearia (Salicaceae) de importância ecológica, econômica e medicinal

The Casearia genus (Salicaceae) is well known because of the medicinal use of its species. Among them, a noteworthy one is the C. sylvestris specie because it already has studies concerning its antiproliferative and/or cytotoxic activity in tumor cells. Furthermore, this specie is popularly used against snake bites, in gastric ulcers treatment and as anti-inflammatory. As well as this, there are other species from this same genus which have been poorly studied, such as the following species: C. decandra, C. grandiflora, C. javitensis, C. arborea, C. lasiophylla and C. ulmifolia. However, several biological activities have been reported for them. In this context, the aim of this project, besides of contributing to the Casearia genus studies, is to study those six species through the analysis and documentation of their leaves' chemical composition (aqueous, ethanolic and hexanic extracts), using analytical separation techniques coupled with spectroscopic techniques, such as UHPLC-DAD, GC-MS and NMR 1H, which will assist the identification of new secondary metabolites in this genus. Moreover, another goal of this present work is aiming the bioprospection of substances with medicinal and economical potential and finally promote the systematic study of some biological activities, such as antimicrobial and cytotoxicity bioassays. A wide variety of metabolites was identified in those three types of extracts, being most of them detected for the first time in Casearia genus, highlighting C. lasiophylla and C. decandra for featuring antimicrobial activity against Staphylococcus aureus.

Documentação química de espécies do gênero Casearia (Salicaceae) de importância ecológica, econômica e medicinal

The Casearia genus (Salicaceae) is well known because of the medicinal use of its species. Among them, a noteworthy one is the C. sylvestris specie because it already has studies concerning its antiproliferative and/or cytotoxic activity in tumor cells. Furthermore, this specie is popularly used against snake bites, in gastric ulcers treatment and as anti-inflammatory. As well as this, there are other species from this same genus which have been poorly studied, such as the following species: C. decandra, C. grandiflora, C. javitensis, C. arborea, C. lasiophylla and C. ulmifolia. However, several biological activities have been reported for them. In this context, the aim of this project, besides of contributing to the Casearia genus studies, is to study those six species through the analysis and documentation of their leaves' chemical composition (aqueous, ethanolic and hexanic extracts), using analytical separation techniques coupled with spectroscopic techniques, such as UHPLC-DAD, GC-MS and NMR 1H, which will assist the identification of new secondary metabolites in this genus. Moreover, another goal of this present work is aiming the bioprospection of substances with medicinal and economical potential and finally promote the systematic study of some biological activities, such as antimicrobial and cytotoxicity bioassays. A wide variety of metabolites was identified in those three types of extracts, being most of them detected for the first time in Casearia genus, highlighting C. lasiophylla and C. decandra for featuring antimicrobial activity against Staphylococcus aureus.

Expanding the Clinical and Genetic Spectrum of Human CD40L Deficiency: The Occurrence of Paracoccidioidomycosis and Other Unusual Infections in Brazilian Patients

CD40 ligand (CD40L) deficiency or X-linked hyper-IgM syndrome (X-HIGM) is a well-described primary immunodeficiency in which Pneumocystis jiroveci pneumonia is a common clinical feature. We have identified an unusual high incidence of fungal infections and other not yet described infections in a cohort of 11 X-HIGM patients from nine unrelated Brazilian families. Among these, we describe the first case of paracoccidioidomycosis (PCM) in X-HIGM. The molecular genetic analysis of CD40L was performed by gene sequencing and evaluation of CD40L protein expression. Nine of these 11 patients (82%) had fungal infections. These included fungal species common to CD40L deficiency (P. jiroveci and Candida albicans) as well as Paracoccidioides brasiliensis. One patient presented with PCM at age 11 years and is now doing well at 18 years of age. Additionally, one patient presented with a simultaneous infection with Klebsiella and Acinetobacter, and one with condyloma caused by human papilloma virus. Molecular analysis revealed four previously described CD40L mutations, two novel missense mutations (c.433 T>G and c.476 G>C) resulting in the absence of CD40L protein expression by activated CD4(+) cells and one novel insertion (c.484_485insAA) within the TNFH domain leading to a frame shift and premature stop codon. These observations demonstrated that the susceptibility to fungal infections in X-HIGM extends beyond those typically associated with X-HIGM (P. jiroveci and C. albicans) and that these patients need to be monitored for those pathogens.

Riabilitazione e valutazione di pazienti con deficit neurologici mediante l'utilizzo di realtà virtuale, feedback aumentati, sensori inerziali e tecnologie video a basso costo.

La neuroriabilitazione è un processo attraverso cui individui affetti da patologie neurologiche mirano al conseguimento di un recupero completo o alla realizzazione del loro potenziale ottimale benessere fisico, mentale e sociale. Elementi essenziali per una riabilitazione efficace sono: una valutazione clinica da parte di un team multidisciplinare, un programma riabilitativo mirato e la valutazione dei risultati conseguiti mediante misure scientifiche e clinicamente appropriate. Obiettivo principale di questa tesi è stato sviluppare metodi e strumenti quantitativi per il trattamento e la valutazione motoria di pazienti neurologici. I trattamenti riabilitativi convenzionali richiedono a pazienti neurologici l’esecuzione di esercizi ripetitivi, diminuendo la loro motivazione. La realtà virtuale e i feedback sono in grado di coinvolgerli nel trattamento, permettendo ripetibilità e standardizzazione dei protocolli. È stato sviluppato e valutato uno strumento basato su feedback aumentati per il controllo del tronco. Inoltre, la realtà virtuale permette l’individualizzare il trattamento in base alle esigenze del paziente. Un’applicazione virtuale per la riabilitazione del cammino è stata sviluppata e testata durante un training su pazienti di sclerosi multipla, valutandone fattibilità e accettazione e dimostrando l'efficacia del trattamento. La valutazione quantitativa delle capacità motorie dei pazienti viene effettuata utilizzando sistemi di motion capture. Essendo il loro uso nella pratica clinica limitato, una metodologia per valutare l’oscillazione delle braccia in soggetti parkinsoniani basata su sensori inerziali è stata proposta. Questi sono piccoli, accurati e flessibili ma accumulano errori durante lunghe misurazioni. È stato affrontato questo problema e i risultati suggeriscono che, se il sensore è sul piede e le accelerazioni sono integrate iniziando dalla fase di mid stance, l’errore e le sue conseguenze nella determinazione dei parametri spaziali sono contenuti. Infine, è stata presentata una validazione del Kinect per il tracking del cammino in ambiente virtuale. Risultati preliminari consentono di definire il campo di utilizzo del sensore in riabilitazione.

Metabolismus von Psychopharmaka durch das Cytochrom P450-Enzymsystem im Alter

Altern geht mit einer Reihe physiologischer Veränderungen einher. Da in höherem Lebensalter überdurchschnittlich viele Arzneistoffe eingenommen werden und häufig mehrere Erkrankungen gleichzeitig vorliegen, können Auffälligkeiten in den Arzneimittelkonzentrationen im Blut nicht nur altersbedingt, sondern auch krankheitsbedingt oder durch Arzneimittelwechselwirkungen verursacht sein.rnrnDie vorliegende Arbeit untersucht die Fragestellung, ob der Arzneimittelmetabolismus bei Alterspatenten generell, oder nur bei Patienten mit Multimorbidität und –medikation verändert ist, und in welchem Lebensalter diese Veränderungen einsetzen. Im Mittelpunkt stand dabei die Frage, ob die Aktivitäten distinkter Arzneimittel-abbauender Enzyme der Cytochrom P450-Enzym-Familie (CYP) verändert sind. Da viele Psychopharmaka nur bei Patienten im Alter zwischen 18 und 65 Jahren zugelassen sind, wurde die Hypothese geprüft, dass sich Patienten im Alter über und unter 65 Jahren in ihren Medikamentenspiegeln unterscheiden.rnrnFür die Untersuchungen wurde eine Datenbank aus Blutspiegelmessungen erstellt, die im Rahmen des pharmakotherapiebegleitenden TDM erhoben worden waren. Die Blutspiegel stammten von insgesamt 4197 Patienten, die mit Amisulprid, Aripiprazol, Citalopram, Clozapin, Donepezil, Escitalopram, Mirtazapin, Quetiapin, Risperidon, Sertralin, Venlafaxin oder Ziprasidon behandelt wurden. Die Messungen wurden ergänzt mit Angaben aus den TDM-Anforderungsscheinen bezüglich Tagesdosis, Begleitmedikamenten, Schweregrad der Erkrankung, Therapieerfolg und Verträglichkeit der Medikation. Zusätzlich wurden klinische Befunde der Leber- und Nierenfunktion einbezogen, sowie Angaben zur Berechnung des BMI. Die in vivo-CYP-Enzymaktivitäten wurden anhand von metabolischen Ratios (Serumkonzentrationen Metabolit/ Serumkonzentration Muttersubstanz) beurteilt.rnrnIm Mittel stieg der Schweregrad der Erkrankung mit dem Alter und der Therapieerfolg verschlechterte sich. Dies betraf im Einzelnen nur Patienten, die mit Amisulprid oder Clozapin behandelt worden waren. Ältere Patienten litten häufiger an Nebenwirkungen als jüngere.rnUnter Aripiprazol, Quetiapin, Sertralin und Venlafaxin erreichten Alterspatienten mit niedrigeren Tagesdosen gleiche Therapieerfolge wie jüngere Patienten.rnPatienten, die mit Clozapin oder Amisulprid behandelt wurden, zeigten im Alter schlechtere Behandlungserfolge bei gleicher (Clozapin) bzw. niedrigerer (Amisulprid) Tagesdosis.rnTherapieerfolg und mittlere Tagesdosis änderten sich bei Patienten, die Ziprasidon, Donepezil, Citalopram, Escitalopram und Mirtazapin einnahmen, nicht altersabhängig.rnrnAltersabhängige Unterschiede der Serumspiegel zeigten sich für Amisulprid, Aripiprazol, Donepezil, Mirtazapin, Desmethylmirtazapin, Quetiapin und DesmethylsertralinrnAllerdings lagen die Altersgrenzen außer bei Donepezil deutlich niedriger als die gängig angenommene, nämlich bei 35 Jahren (Aripiprazol), 70 Jahren (Donepezil), 55 Jahren (D-Sertralin), 41 Jahren (Amisulprid), 49 Jahren (Quetiapin) und 58 Jahren (Mirtazapin).rnEs bestand kein Zusammenhang zwischen dem Auftreten veränderter Serumspiegel im Alter und dem Verteilungsvolumen, der Plasmaproteinbindung oder der Eliminationshalbwertszeit der untersuchten Wirkstoffe.rnrnBei Patienten ohne Comedikation fand sich in keinem Fall eine altersabhängige Veränderung der Ratio. Es ergab sich daher kein Hinweis auf eine Veränderung der CYP-Aktivität im Alter. Die Einnahme von Comedikation nahm mit dem Alter zu, hierfür ließ sich eine Altersgrenze von 49 Jahren definieren. Unter Polytherapie wurden Veränderungen der CYP-Aktivität beobachtet.rnrnDer Einfluss veränderter Leber- oder Nierenfunktion auf die Biotransformation von Pharmaka wurde anhand von Serumspiegeln von Patienten, die mit Donepezil, Venlafaxin, Citalopram oder Escitalopram behandelt wurden, untersucht. rnBei keinem Wirkstoff wurden unter auffälligen Leber- oder Nierenparametern signifikant veränderte Serumspiegel gemessen.rnEine Abhängigkeit der Serumspiegel vom Körpergewicht wurde nur für Desmethylsertralin gefunden. Die Spiegel waren bei Patienten mit einem Body Mass Index unter 20 signifikant höher als bei Patienten mit einem Index über 20. Aufgrund der kleinen Fallgruppe und der Tatsache, dass der Serumspiegel der Muttersubstanz nicht stieg, konnte nicht zwingend von einem Alterseinfluss aufgrund der veränderten Körperzusammensetzung ausgegangen werden.rnInsgesamt ergaben sich aus den Untersuchungen Hinweise auf moderate altersabhängige Veränderungen der Pharmakokinetik. Es ließen sich allerdings keine allgemeinen Dosierempfehlungen für Alterspatienten ableiten. Es zeigte sich jedoch, dass mit altersabhängigen Veränderungen der Pharmakokinetik bereits nach dem 50. Lebensjahr zu rechnen ist. Weitere Untersuchungen sollten auch den Alterseffekt auf gastrointestinale Transporter einbeziehen, die die aktive Aufnahme von Arzneistoffen ins Blut bewerkstelligen. Unklar ist auch die Rolle des Alterns auf die Aktivität des P-Glykoproteins. rn

Ctenidins: antimicrobial glycine-rich peptides from the hemocytes of the spider Cupiennius salei

Three novel glycine-rich peptides, named ctenidin 1-3, with activity against the Gram-negative bacterium E. coli, were isolated and characterized from hemocytes of the spider Cupiennius salei. Ctenidins have a high glycine content (>70%), similarly to other glycine-rich peptides, the acanthoscurrins, from another spider, Acanthoscurria gomesiana. A combination of mass spectrometry, Edman degradation, and cDNA cloning revealed the presence of three isoforms of ctenidin, at least two of them originating from simple, intronless genes. The full-length sequences of the ctenidins consist of a 19 amino acid residues signal peptide followed by the mature peptides of 109, 119, or 120 amino acid residues. The mature peptides are post-translationally modified by the cleavage of one or two C-terminal cationic amino acid residue(s) and amidation of the newly created mature C-terminus. Tissue expression analysis revealed that ctenidins are constitutively expressed in hemocytes and to a small extent also in the subesophageal nerve mass.

Phenotype-genotype correlation in eight Polish patients with inherited Factor XIII deficiency: identification of three novel mutations

Inherited factor XIII (FXIII) deficiency is known as one of the most rare blood coagulation disorder in humans. In the present study, phenotype and genotype of eight FXIII deficient Polish patients from five unrelated families were compared. The patients presented with a severe phenotype demonstrated by a high incidence of intracerebral haemorrhages (seven of eight patients), haemarthrosis (six patients) and bleeding due to trauma (five patients). Introduction of regular substitution with FXIII concentrate prevented spontaneous bleeding in seven patients. In all patients, mutations within the F13A gene have been identified revealing four missense mutations (Arg77Cys, Arg260Cys, Ala378Pro, Gly420Ser), one nonsense mutation (Arg661X), one splice site mutation (IVS5-1 G>A) and one small deletion (c.499-512del). One homozygous large deletion involving exon 15 was detected by failure of PCR product. The corresponding mutations resulted in severely reduced FXIII activity and FXIII A-subunit antigen concentration, while FXIII B-subunit antigen remained normal or mildly decreased. Structural analysis demonstrated that the novel Ala378Pro mutation may cause a disruption of the FXIII catalytic triad leading to a non-functional protein which presumably undergoes premature degradation. In conclusion, the severe phenotype with high incidence of intracranial bleeding and haemarthrosis was in accordance with laboratory findings on FXIII and with severe molecular defects of the F13A gene.

Allocentric visual cues influence mental transformation of bodies

Identifying a human body stimulus involves mentally rotating an embodied spatial representation of one's body (motoric embodiment) and projecting it onto the stimulus (spatial embodiment). Interactions between these two processes (spatial and motoric embodiment) may thus reveal cues about the underlying reference frames. The allocentric visual reference frame, and hence the perceived orientation of the body relative to gravity, was modulated using the York Tumbling Room, a fully furnished cubic room with strong directional cues that can be rotated around a participant's roll axis. Sixteen participants were seated upright (relative to gravity) in the Tumbling Room and made judgments about body and hand stimuli that were presented in the frontal plane at orientations of 0°, 90°, 180° (upside down), or 270° relative to them. Body stimuli have an intrinsic visual polarity relative to the environment whereas hands do not. Simultaneously the room was oriented 0°, 90°, 180° (upside down), or 270° relative to gravity resulting in sixteen combinations of orientations. Body stimuli were more accurately identified when room and body stimuli were aligned. However, such congruency did not facilitate identifying hand stimuli. We conclude that static allocentric visual cues can affect embodiment and hence performance in an egocentric mental transformation task. Reaction times to identify either hands or bodies showed no dependence on room orientation.

Lobe specific Ca2+-calmodulin nano-domain in neuronal spines: a single molecule level analysis.

Calmodulin (CaM) is a ubiquitous Ca(2+) buffer and second messenger that affects cellular function as diverse as cardiac excitability, synaptic plasticity, and gene transcription. In CA1 pyramidal neurons, CaM regulates two opposing Ca(2+)-dependent processes that underlie memory formation: long-term potentiation (LTP) and long-term depression (LTD). Induction of LTP and LTD require activation of Ca(2+)-CaM-dependent enzymes: Ca(2+)/CaM-dependent kinase II (CaMKII) and calcineurin, respectively. Yet, it remains unclear as to how Ca(2+) and CaM produce these two opposing effects, LTP and LTD. CaM binds 4 Ca(2+) ions: two in its N-terminal lobe and two in its C-terminal lobe. Experimental studies have shown that the N- and C-terminal lobes of CaM have different binding kinetics toward Ca(2+) and its downstream targets. This may suggest that each lobe of CaM differentially responds to Ca(2+) signal patterns. Here, we use a novel event-driven particle-based Monte Carlo simulation and statistical point pattern analysis to explore the spatial and temporal dynamics of lobe-specific Ca(2+)-CaM interaction at the single molecule level. We show that the N-lobe of CaM, but not the C-lobe, exhibits a nano-scale domain of activation that is highly sensitive to the location of Ca(2+) channels, and to the microscopic injection rate of Ca(2+) ions. We also demonstrate that Ca(2+) saturation takes place via two different pathways depending on the Ca(2+) injection rate, one dominated by the N-terminal lobe, and the other one by the C-terminal lobe. Taken together, these results suggest that the two lobes of CaM function as distinct Ca(2+) sensors that can differentially transduce Ca(2+) influx to downstream targets. We discuss a possible role of the N-terminal lobe-specific Ca(2+)-CaM nano-domain in CaMKII activation required for the induction of synaptic plasticity.