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Sensory and methodological aspects in biomechanical research of postural control and clinical fields of applicationSchmidt, Daniel 23 February 2022 (has links)
The human senses constitute a highly complex system based on various sensory organs, afferent pathways, and central processing locations, which allow us to
interact with the environment, but also with ourselves. A further domain is important to achieve this interaction: the motor system, which allows linguistic communication
and locomotion, for example. It becomes evident that sensory receptors work as a source of information to initiate, optimize, or cease motor activity. One generic term
for such sensory sources is the somatosensory system, which is mainly based on receptors located in muscles, tendons, and the skin (cutaneous sensitivity). In this
regard, it has been shown that cutaneous sensitivity contributes to human balance regulation. However, there are still debates concerning the exact role of plantar (foot
sole) receptor inputs in particular, and how their isolated contribution to, e.g., balance regulation may be assessed accordingly.
To investigate the interaction between plantar cutaneous sensitivity and human balance capabilities, several aspects need to be considered which are still
controversial and inconclusive in the scientific community. For example, when assessing cutaneous vibration sensitivity, it is well-known that increasing vertical
forces of the contactor toward the skin usually result in improved sensitivity.
However, it has not been profoundly investigated whether assessing plantar vibratory sensitivity differs when comparing a standing or sitting posture, which obviously involves different contactor forces. In addition, many studies implementing cutaneous sensitivity show certain limitations with respect to adequate data analyses. A similar aspect also applies when assessing balance performance: devices allowing an investigation of dynamic balance performance (induced by unexpected platform perturbations while standing, for example) have only been partially investigated with regard to their biomechanical quality criteria, such as reliability.
With these considerations in mind, the present doctoral thesis is based on five published studies. Study 1 investigates if plantar sensitivity is influenced by different
body positions when collecting data. Study 2 asks how to appropriatly analze plantar sensitivity data. Study 3 examines the reliability of dynamic balance responses using the so-called Posturomed device, and Study 4 identifies the isolated role of plantar inputs on balance responses, when an acute sensory manipulation is induced that exclusively affects plantar aspects. Ultimately, clinical fields of application (based on the previous four studies) are highlighted in Study 5.
The main findings of the first four studies can be summarized as follows. First, higher contact forces when standing compared to sitting did not influence plantar sensitivity. This is an important finding, as plantar sensitivity tests (often performed during sitting) may, hence, be brought into context with balance tests usually performed during standing. Second, plantar sensitivity data are shown to exhibit heteroscedasticity, meaning that the measurement error increases as the values increase. In Study 2, we provided an easy-to-follow example for how to account for
heteroscedasticity by logarithmizing the raw data, and how to control whether this approach was successful in eliminating heteroscedasticity. Third, dynamic balance
responses assessed via the Posturomed device exhibit an overall good reliability.
Occasional significant differences were shown to be clinically non-relevant, identified by root mean square error calculations. Fourth, a permanent plantar sensory
manipulation (hypothermia) was successfully achieved and maintained throughout data collection. Study 4 showed that the reduced plantar sensory input due to the
hypothermic manipulation was compensated during more unchallenging balance conditions (standing still). There was no full compensation during more challenging
balance conditions (unexpected platform perturbations during standing), however, with the body reacting with cautious motor behavior. This became evident by
decreased outcome measures following hypothermic plantar sensory manipulation.
These four studies shed further light onto investigations combining sensory and motor tests, especially with regard to physiological and methodological aspects that
should be considered when analyzing and interpreting associated data.
Finally, this doctoral thesis also provides an example of identifying clinical fields of application concerning sensory-focused research. In Study 5, we highlight the role of sensory research in the (early) diagnosis of diseases associated with cognitive decline. For this purpose, various instruments such as sensory tests or coordinative motor tests are implemented. Preliminary results suggest that not only classical cognitive parameters and questionnaires should be used to identify and better understand cognitive decline. / Die menschlichen Sinne stellen ein sehr komplexes System dar, welches auf verschiedenen sensorischen Organen, afferenten Leitungsbahnen und zentralen Verarbeitungsstellen basiert und es uns ermöglicht, mit der Umwelt, aber auch mit uns selbst, zu interagieren. Dahingehend ist eine weitere wichtige Domäne wichtig, um diese Interaktion zu bewerkstelligen: das motorische System, welches etwa eine sprachliche Kommunikation oder auch die Fortbewegung ermöglicht. Es wird somit offensichtlich, dass sensorische Rezeptoren eine Informationsquelle darstellen, um motorische Aktivität zu initiieren, zu optimieren oder zu beenden. Ein grundlegender Terminus für solch sensorische Quellen ist das somatosensorische System, welches überwiegend auf Rezeptoren in Muskulatur, Sehnen und der Haut (kutane Sensibilität) beruht. Diesbezüglich wurde bereits aufgezeigt, dass die kutane Sensibilität einen Beitrag bei der menschlichen Gleichgewichtsregulation leistet.
Allerdings existieren dabei nachwievor Diskussionen in Bezug auf die genaue Bedeutung plantarer (die Fußsohle betreffend) Rezeptor-Inputs und inwieweit deren
isolierte Bedeutung bei der Gleichgewichtsregulation entsprechend ermittelt werden kann.
Um die Interaktion zwischen der kutanen Sensorik der Fußsohle und der menschlichen Gleichgewichtsfähigkeit zu erforschen, sollten verschiedene Aspekte
berücksichtigt werden, welche nachwievor kontrovers und nicht eindeutig in der Wissenschaft diskutiert werden. Bei Erhebungen der kutanen Vibrationssensibilität,
als Beispiel, ist bereits bekannt, dass erhöhte Vertikalkräfte, mit denen der Vibrationsstößel gegen die Haut appliziert ist, generell zu einer verbesserten Sensibilität/Sensorik führen. Allerdings wurde noch nicht klar erforscht, ob sich die plantare Vibrationssensibilität zwischen einer stehenden und sitzenden Haltung der Probanden/innen unterscheidet, wobei hier natürlich unterschiedliche Vertikalkräfte der Stößel wahrscheinlich sind. Darüber hinaus zeigen viele Studien, welche die Hautsensibilität untersuchen, gewisse Limitierungen in Bezug auf eine adäquate Datenanalyse. Ein sehr ähnlicher Aspekt trifft auch auf die Evaluierung der Gleichgewichtsfähigkeit zu: Messgeräte, welche dabei eine Erfassung der dynamischen Gleichgewichtsfähigkeit zulassen (z.B. eingeleitet durch unerwartete Plattform-Perturbationen während des Stehens), wurden bisher nur teilweise auf die biomechanischen Gütekriterien hin untersucht, wie etwa die Reliabilität.
Aufgrund dieser Überlegungen basiert die vorliegende Dissertation auf fünf publizierten Studien, welche folgende Aspekte untersuchten: Wird die plantare Sensibilität durch verschiedene Körperpositionen während der Datenaufnahme beeinflusst (Studie 1)? Wie können plantare Sensibilitätsdaten angemessen analysiert werden (Studie 2)? Darüber hinaus wurde ebenso untersucht, inwiefern das sogenannte 'Posturomed'-Messgerät bei der Beurteilung dynamischer Gleichgewichtsantworten reliable Messwerte liefert (Studie 3). Ferner wurde in Studie
4 untersucht, inwiefern isoliert plantare Inputsignale bei Gleichgewichtsantworten relevant sind (anhand einer akuten sensorischen Manipulation, welche ausschließlich
die Fußsohle betrifft). In Studie 5 werden konkrete klinische Anwendungsbeispiele aufgrund der vier hier vorgestellten Studien aufgezeigt.
Die Hauptergebnisse der ersten vier Studien können wie folgt zusammengefasst werden: Erstens, höhere vertikale Kontaktkräfte während des Stehens verglichen mit sitzenden Positionen führten zu keinen Unterschieden bzgl. der plantaren Sensibilität. Dies ist eine wichtige Erkenntnis, da plantare Sensorikmessungen (oft während des Sitzens durchgeführt) dadurch in Kontext mit Gleichgewichtstests gebracht werden können, welche normalerweise im Stehen erfolgen. Zweitens, Daten der plantaren Sensorik zeigten Heteroskedastizität, was bedeutet, dass sich der Messfehler mit Größenzunahme der Messwerte ebenso erhöht. Wir konnten in Studie 2 ein leicht zu erschließendes Beispiel aufzeigen, wie das Problem der Heteroskedastizität durch eine Logarithmierung der Rohdaten behandelt werden konnte und wie kontrolliert werden konnte, ob diese Behandlung erfolgreich war. Drittens, die dynamischen Gleichgewichtsantworten, welche
mittels des 'Posturomed' ermittelt wurden, zeigen insgesamt eine gute Reliabilität. Gelegentlich auftretende signifikante Unterschiede wurden anhand von Berechnungen der Wurzel
der mittleren Fehlerquadratsumme (root mean square error, RMSE) als klinisch nicht relevant eingestuft. Viertens, eine anhaltende plantar-sensorische Manipulation (Hypothermie) wurde erfolgreich eingeleitet und während der Datenerhebung aufrecht erhalten. Studie 4 zeigte ferner, dass die hypothermisch eingeleiteten reduzierten plantaren Sensorik-Inputs während der eher nicht herausfordernden quasi-statischen Gleichgewichtsbedingungen (einfaches aufrechtes Stehen) kompensiert werden konnten. Während der herausfordernden Gleichgewichtskonditionen (unerwartete Perturbationen der Plattform während des Stehens) hingegen wurde keine vollständige Kompensation erreicht. Allerdings reagierten die Probanden mit einem vorsichtigen motorischen Verhalten. Dies wurde durch die reduzierten Ergebnisparameter infolge der plantaren hypothermischen Manipulation ersichtlich. Die vier hier genannten Studien zeigen weitere Erkenntnisse in Bezug auf Forschungsaktivitäten, welche sensorische und motorische Tests vereinen. Dies trifft speziell in Hinblick auf physiologische und methodologische
Aspekte zu, welche bei der Analyse und Interpretation derartiger Daten in Betracht gezogen werden sollten. Zuletzt bietet diese Arbeit auch ein Beispiel dafür, welche klinischen Anwendungsfelder im Bereich der sensorisch-fokussierten Forschung identifiziert werden können. In Studie 5 wird dafür die Bedeutung sensorischer Forschung bei der (Früh-) Diagnose von Erkrankungen aufgezeigt, welche mit kognitiven Einschränkungen in Verbindung gebracht werden. Für diesen Zweck werden verschiedene Instrumente eingebracht, wie etwa sensorische oder koordinativ-motorische Tests. Vorläufige Ergebnisse deuten dabei bereits an, dass nicht nur die klassischen kognitiven Parameter und Fragebögen bei der Identifizierung oder zum Zwecke des besseren Verstehens kognitiven Verfalls einbezogen werden sollten.
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Contribution à l’évaluation de la toxicocinétique humaine du bisphénol SKhmiri, Imen 10 1900 (has links)
La mesure du bisphénol-S (BPS) et de son glucurono-conjugué (BPSG) dans l’urine peut être utilisée pour la biosurveillance de l’exposition dans les populations. Cependant, cela nécessite une connaissance approfondie de la toxicocinétique de ces composés alors qu’à ce jour, il existe peu de données à cet effet chez l’humain. L’évolution dans le temps du BPS et du BPSG a été évaluée dans des matrices biologiques accessibles et représentatives comme l`urine et le sang de volontaires exposés par voie orale et cutanée. Suite à l’approbation du comité d’éthique de la recherche de l’Université de Montréal, six volontaires ont été exposés par voie orale à une dose deutérée de BPS-d8 de 0,1 mg/kg de poids corporel. Un mois plus tard, 1 mg/kg pc de BPS-d8 ont été appliqués sur 40 cm2 de l’avant-bras puis lavés 6 h après l’application. Des échantillons de sang ont été prélevés avant le dosage et à des intervalles de temps fixes sur une période de 48 h après traitement ; des collectes urinaires complètes ont été recueillies avant l’exposition et à des intervalles préétablis sur 72 h après dosage.
Après exposition par voie orale, les profils temporels des concentrations plasmatiques de BPS-d8 et de BPSG-d8 évoluaient en parallèle et ont montré une apparition et une élimination rapides. Les valeurs maximales de BPS-d8 et BPSG-d8 dans le plasma ont été atteintes en moyenne (± écart-type [ET]) à 0,7 ± 0,1 et 1,1 ± 0,4 h après le dosage et les demi-vies d’élimination apparentes (moyenne ± ET) (t ½) de 7,9 ± 1,1 et 9,3 ± 7,0 h ont été calculées à partir de la phase terminale, respectivement. La fraction de BPS-d8 atteignant la circulation systémique inchangée (c’est-à-dire la biodisponibilité) a en outre été estimée à 62 ± 5 % en moyenne (± ET) et la clairance plasmatique systémique à 0,57 ± 0,07 L/kg pc/h. Toujours après exposition orale, les profils temporels des taux d’excrétion urinaire évoluaient aussi de manière parallèle aux concentrations plasmatiques et étaient similaires pour tant pour le composé parent que le métabolite. Le pourcentage moyen (± ET) de la dose administrée récupérée dans l’urine sous forme de BPS-d8 et BPSG-d8 au cours de la période de 72 h après le dosage était de 1,72 ± 1,3 et 54 ± 10 %.
Après application cutanée, les niveaux plasmatiques étaient inférieurs à la limite inférieure de quantification (LLOQ) à la plupart des points dans le temps. Cependant, les valeurs maximales étaient atteintes entre 5 et 8 h selon les individus, suggérant un taux d’absorption plus lent par rapport à l’exposition orale. De même, des quantités limitées de BPS-d8 et de son conjugué, estimées en pourcentage de dose, de l’ordre 0,004 ± 0,003 et 0,09 ± 0,07 %.
En somme, cette étude a fourni une plus grande précision sur la cinétique du BPS chez l’humain. Ces données seront utiles pour développer un modèle toxicocinétique pour une meilleure interprétation des données de biosurveillance. Pour la voie orale, le taux d’absorption apparent similaire du BPS-d8 et BPSG-d8 après une exposition par voie orale suggère que les formes libres et conjuguées du BPS atteignent la circulation sanguine systémique à peu près dans le même intervalle de temps. Ceci indique un effet de premier passage hépatique, c’est-à-dire une conjugaison du BPS-d8 dans le foie avant d’atteindre la circulation systémique. Néanmoins, malgré l’effet de premier passage, la biodisponibilité du BPS-d8 et donc la forme active non conjuguée du BPS dans le sang est relativement élevée. Celle-ci est en fait largement plus importante que celle du BPA. Le BPS libre s’est aussi avéré avoir a un temps de résidence plasmatique plus long que le BPA. Pour la voie cutanée, les données ont montré que le BPS atteignait rapidement la circulation sanguine systémique et était donc rapidement absorbé par peau. Par contre, son élimination du corps semble être plus lente après exposition cutanée comparativement à son élimination après exposition orale. Par ailleurs, malgré le fait que le BPS serait rapidement absorbé au niveau de la peau, la fraction d’absorption cutanée était très faible par rapport à la fraction d’absorption orale. / The measurement of bisphenol-S (BPS) and its glucurono-conjugate (BPSG) in urine may be used for the biomonitoring of exposure in populations. However, this requires a thorough knowledge of their toxicokinetics. The time courses of BPS and BPSG were assessed in accessible biological matrices of orally and dermally exposed volunteers. Under the approval of the Research Ethics Committee of the University of Montreal, six volunteers were orally exposed to a BPS-d8 deuterated dose of 0.1 mg/kg body weight (bw). One month later, 1 mg/kg bw of BPS-d8 were applied on 40 cm2 of the forearm and then washed 6 h after application. Blood samples were taken prior to dosing and at fixed time periods over 48 h after treatment; complete urine voids were collected pre-exposure and at pre-established intervals over 72 h postdosing.
Following oral exposure, the plasma concentration–time courses of BPS-d8 and BPSG-d8 over 48 h evolved in parallel and showed a rapid appearance and elimination. Average peak values (±SD) were reached at 0.7 ± 0.1 and 1.1 ± 0.4 h postdosing and mean (±SD) apparent elimination half-lives (t½) of 7.9 ± 1.1 and 9.3 ± 7.0 h were calculated from the terminal phase of BPS-d8 and BPSG-d8 in plasma, respectively. The fraction of BPS-d8 reaching the systemic circulation unchanged (i.e. bioavailability) was further estimated at 62 ± 5% on average (±SD) and the systemic plasma clearance at 0.57 ± 0.07 L/kg bw/h. Plasma concentration–time courses and urinary excretion rate profiles roughly evolved in parallel for both substances, as expected. The average percent (±SD) of the administered dose recovered in urine as BPS-d8 and BPSG-d8 over the 0–72 h period postdosing was 1.72 ± 1.3 and 54 ± 10%.
Following dermal application, plasma levels were under the lower limit of quantification (LLOQ) at most time points. However, peak values were reached between 5 and 8 h depending on individuals, suggesting a slower absorption rate compared to oral exposure. Similarly, limited amounts of BPS-d8 and its conjugate were recovered in urine and peak excretion rates were reached between 5 and 11 h postdosing. The average percent (±SD) of the administered dose recovered in urine as BPS-d8 and BPSG-d8 was about 0.004 ± 0.003 and 0.09 ± 0.07%, respectively.
This study provided greater precision on the kinetics of this contaminant in humans and, in particular, evidenced major differences between BPA and BPS kinetics with much higher systemic levels of active BPS than BPA, an observation explained by a higher oral bioavailability of BPS than BPA. These data should also be useful in developing a toxicokinetic model for a better interpretation of biomonitoring data.
Overall, this study provided greater precision on the kinetics of this contaminant in humans. These data will be useful in developing a toxicokinetic model for a better interpretation of biomonitoring data. For the oral route of exposure, the apparent similar absorption rate of BPS-d8 and BPSG-d8 after oral exposure suggests that free and conjugated forms of BPS reach the systemic blood circulation at about the same time interval. This indicates a first-pass effect in the liver, i.e. a conjugation of BPS-d8 in the liver before reaching the systemic circulation. Nevertheless, despite the first-pass effect, the bioavailability of BPS-d8 and therefore the proportion of unconjugated active form of BPS reaching the systemic bloodstream is relatively high. It is actually much higher than that of BPA. Free BPS was also found to have a longer plasma residence time than BPA. For the dermal route of exposure, data show that BPS quickly reaches systemic blood circulation and is therefore rapidly absorbed by skin. On the other hand, its elimination from the body appears to be slower after dermal exposure compared to its elimination after oral exposure. Furthermore, despite the fact that BPS is rapidly absorbed through the skin, the dermal absorption fraction was very small compared to the oral absorption fraction.
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In Vitro Percutaneous Absorption Studies of Cannabidiol Using Human Skin: Exploring the Effect of Drug Concentration, Chemical Enhancers, and Essential OilsJunaid, Mohammad S., Tijani, Akeemat O., Puri, Ashana, Banga, Ajay K. 25 March 2022 (has links)
Cannabidiol, a non-psychoactive constituent of cannabis, has garnered much attention after United States Food and Drug Administration approved Epidiolex® for oral use. Although therapeutic effect of cannabidiol after systemic absorption has been investigated extensively, its therapeutic potential in treating skin disorders after local delivery still needs further exploration. Our study has investigated the effect of cannabidiol concentration, chemical enhancers, and essential oils on percutaneous absorption of cannabidiol. In vitro permeation tests were conducted on human skin. The 24 h study results suggest no significant difference in amount of drug absorbed into skin, between 5% (242.41 ± 12.17 µg/cm) and 10% (232.79 ± 20.82 cm) cannabidiol solutions. However, 1% delivered (23.02 ± 4.74 µg/cm) significantly lower amount of drug into skin than 5% and 10%. Transcutol and isopropyl myristate did not enhance delivery of cannabidiol. However, oleic acid was found to be useful as chemical enhancer. Oleic acid (43.07 ± 10.11 µg/cm) had significantly higher cannabidiol delivery into skin than the group without oleic acid (10.98 ± 3.40 µg/cm) after a 4 h in vitro permeation study. Essential oils at concentrations tested had lower total cannabidiol delivery when compared to control. This study's findings will help guide future research on the pharmacological effect of percutaneously delivered cannabidiol on inflammatory skin disorders.
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Manipulation of space and time in the tactile universeDeep, Akash 23 November 2018 (has links)
The study of tactile illusions like visual illusions can reveal the brain's processing strategies. A famous tactile illusion is the cutaneous rabbit illusion. Fundamental to this illusion is the perceptual length contraction phenomenon: two taps that occur in rapid succession on the forearm are perceived as occurring closer together than they were physically placed. Our lab previously proposed a Bayesian probabilistic model that views perception as a compromise between expectation (prior experience) and sensation (likelihood of sensorineural data given hypothesized tap locations). The model proposes a low-speed prior, an expectation based on experience that objects tend to be stationary or to move slowly on the skin. When the sensation of space is unclear (e.g., taps are weak), the model predicts that expectation will strongly influence perception. Consistent with this prediction, our lab previously showed that the use of weaker taps causes more pronounced perceptual length contraction. Here we report psychophysical tests on 64 participants, which confirmed this finding. Our study also used stimulus sequences consisting of a weak and a strong tap, for which the Bayesian model predicts an asymmetric perceptual length contraction, such that the weaker tap location will be perceived to shift more than the stronger tap. The experimental results confirmed this prediction, providing further support for our Bayesian probabilistic model as an explanation for perceptual length contraction. However, our results revealed a discrepancy in the data at the smaller SOAs, which showed less length contraction than predicted. We hypothesized that participants might overestimate the smaller SOAs, an effect our lab defines as time dilation. Accordingly, in a second study we investigated the effects of varying SOA and lengths on perceived SOA. The model predicts more pronounced time dilation at smaller SOAs and larger lengths. The psychophysical data from 37 participants confirmed the trends predicted by the model. / Thesis / Master of Science (MSc)
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Características sociodemográficas y epidemiológicas de pacientes con cáncer de piel diagnosticados en un Hospital Nivel III-1 de región Lambayeque 2016-2019Rufasto Ñañez, Claudia Estefany January 2024 (has links)
Objetivo: Identificar las características sociodemográficas y epidemiológicas del paciente con cáncer de piel diagnosticados en el servicio de anatomía patológica del Hospital Regional Lambayeque durante periodo enero del 2016 - diciembre del 2019. Métodos: La metodología empleada durante esta investigación estuvo basada en el diseño no experimental, descriptivo, de carácter retrospectivo, trasversal y observacional. Se incluyeron un total de 429 pacientes mayores de 18 años, diagnosticados con carcinoma cutáneo de tipo no melanoma (NPNM) y melanoma, mediante estudios anatomopatológicos de la lesión atendidos en Hospital Regional Lambayeque. La elección de la muestra fue mediante un muestreo no probabilístico de tipo censal, por la adaptabilidad al estudio. Resultados: De un total de 429 pacientes, 256(59,1%) tenían carcinoma basocelular (CBC), 146 (33,7%) carcinoma epidermoide (CsCC) y 31(7,2%) melanoma maligno cutáneo (MM). Siendo la edad promedio de aparición de 71 años en los NPNM y 62 años en el Melanoma Maligno Cutáneo, con predominio por el sexo femenino en el CBC y masculino en CsCC y MM. La ubicación anatómica más comprometida fue de la
cabeza en los NPNM y miembros inferiores en MM, los cuales fueron identificadas mayormente por el servicio de Dermatología, seguido por Cirugía de Cabeza y Cuello del hospital. Los años con mayor número de carcinomas cutáneos fueron el 2019 para CBC y 2018 para los dos restantes. Conclusiones: La población general presenta más riesgo de presentar carcinomas no melanómico y en menor número el melanoma maligno, el cual predomina en áreas fotoexpuestas del cuerpo. / Objective: To identify the sociodemographic and epidemiological characteristics of the patient with skin cancer diagnosed in the pathological anatomy service of the Lambayeque Regional Hospital during the period January 2016 - December 2019. Methods: The methodology used during this investigation was based on the non-experimental design, descriptive, retrospective, cross-sectional and observational. A total of 429 patients over 18 years of age were included, diagnosed with non-melanoma skin carcinoma (NPNM) and melanoma, through anatomopathological studies of the lesion treated at Hospital Regional
Lambayeque. The selection of the sample was by means of a non-probabilistic sampling of the census type, due to the adaptability to the study. Results: Of a total of 429 patients, 256 (59.1%) had basal cell carcinoma (BCC), 146 (33.7%) squamous cell carcinoma (SCC) and 31 (7.2%) cutaneous malignant melanoma (MM). Being the average age of appearance of 71 years in NPNM and 62 years in Cutaneous Malignant Melanoma, with a predominance of females in CBC and male in CsCC and MM. The most compromised anatomical location was the head in NPNM and lower limbs in MM, which were mostly identified by the Dermatology service, followed by Head and Neck Surgery at the hospital. The years with the highest number of skin carcinomas were 2019 for CBC and 2018 for the remaining two. Conclusions: The general population presents a higher risk of presenting non-melanoma carcinomas and a smaller number of malignant melanoma, which predominates in photo-exposed areas of the body.
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"Avaliação da reação em cadeia de polimerase (PCR) no diagnóstico da leishmaniose cutânea no Estado do Espírito Santo, Brasil" / Evaluation of polymerase chain reaction (PCR) in the diagnosis of cutaneous leishmaniosis in the Espírito Santo state, BrazilPassos, Luciana Neves 09 October 2003 (has links)
A identificação de espécie de Leishmania é crucial no diagnóstico de leishmaniose cutânea (LC), para terapia e prognóstico, em áreas com diferentes espécies endêmicas. A reação em cadeia de polimerase (PCR) foi usada em amostras de biópsias estocadas em parafina e formol de pacientes com LC, do Espírito Santo, Brasil. Usando sequências de mini-exon do gênero Leishmania, 63,2% (36/57) das amostras em parafina e 46,1% (6/13) das amostras em formol foram positivas. Numa abordagem usando sequências de kDNA e RFLP, 100% (58/58) das amostras de parafina testadas foram identificadas como Leishmania (V.) braziliensis. Estas reações podem ser usadas tanto para diagnóstico como para definição da espécie infectante / The identification of Leishmania species, a crucial step in cutaneous leishmaniasis, had therapeutics and prognostics implications, specially at when several agent species are endemic. Polymerase chain reaction (PCR) was used for analysis of paraffin-embedded and formalin stored skin biopsies from patients with parasitologic confirmed cutaneous formalin stored skin biopsies from patients with parasitologic confirmed cutaneous leishmaniasis, from the Espirito Santo State, Brazil. Tested by PCR targeted to mini-exon genus specific primer, 63,2% (36/57) of parafin and 46,1% (6/13) of formalin stored samples were positive, but using a PCR for kDNA primer followed by RFLP analysis, only Leishamnia (V.) braziliensis was identified in 100% (58/58) of parafin biopsies studied. Those reactions allow either diagnosis or species identification in cutaneous leismaniasis
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Molecular approaches to direct diagnosis and characterization of Leishmania donovani in clinical isolatesTai, Nahla Omer Ahmed El 06 March 2003 (has links)
Die vorliegende Studie wurde in einer Gruppe von Dörfern im Ostsudan, Gedaref State, durchgeführt. Bei 100 Patienten mit der Verdachtsdiagnose Kala Azar- oder Post-Kala Azar- Leishmaniose war der Erregernachweis mit der PCR direkt in klinischen Proben, die auf Filterpapier aufgebracht worden waren, ohne vorherige Kultivierung erfolgreich. In dieser PCR wurden die ribosomalen, internal transcribed spacer (ITS1 & ITS2) amplifiziert, weil sie sehr variabel sind, eine klare Speziesidentifizierung gestatten und bei weiterführenden Analysen der PCR-Produkte auch der Nachweis stammspezifischer Unterschiede erwartet werden konnte. Für die Analyse der Diversität von Leishmania donovani-Isolaten aus dem Sudan wurden 4 verschiedene PCR-basierte Methoden eingesetzt: das PCR-Fingerprinting mit unspezifischen Einzelprimern, die RFLP- Analyse des amplifizierten ITS-Locus, ,,single strand conformation polymorphism (SSCP)- Analysen der amplifizierten ITS-Region, des Gens, welches für die Hauptoberflächenprotease (gp63) kodiert, und anonymer DNA- Fragmente sowie Sequenzanalysen der entsprechenden Zielregionen. Das PCR-Fingerprinting und die Restriktionsanalyse der ITS-Region lieferten weitgehend übereinstimmende Fragmentmuster für alle untersuchten L.donovani-Stämme. 12 unterschiedliche Profile wurden bei der SSCP-Analyse des ITS1-Locus für 86 Isolate aus dem Sudan erhalten, während der ITS2-Locus bei diesen Stämmen hochkonserviert war und nur ein Stamm ein unterschiedliches SSCP-Muster aufwies. L. Donovani -Stämme anderer geographischer Herkunft hatten unterschiedliche ITS2-Profile in der SSCP. Für den gp63 - Locus waren 3 polymorphe SSCP-Muster bei 31 untersuchten sudanesischen Isolaten nachweisbar. Für die meisten der anonymen DNA-Fragmente, L510, L413, LK413, L0308 UND L0114, konnten leider nur von 8 kultivierten Stämmen gute PCR-Produkte erhalten werden. Lediglich das Fragment L0110 konnte erfolgreich von 31 auf Filterpapier aufgebrachten Proben direkt amplifiziert werden. Die Suche nach Polymorphismen mit der SSCP ergab keine Unterschiede in diesen anonymen DNA-Regionen, mit Ausnahme des Fragments L0114, das zwei verschiedene Muster aufwies. Die Ergebnisse der SSCP-Analysen und der DNA- Sequenzierung stimmten gut überein, wodurch bestätigt wurde, dass die SSCP genetische Unterschiede auf dem Niveau einzelner Basenaustausche nachweisen kann. Die SSCP- Technik hat Vorteile gegenüber den anderen Methoden, die für die Untersuchung von Sequenzvariationen innerhalb der Spezies L. donovani angewandt wurden. Es konnten keine Korrelationen zwischen der Form der klinischen Manifestation und den Ergebnissen des PCR- Fingerprinting, der ITS-RFLP- und ITS-SSCP- Analysen festgestellt warden. Diese Studie ist von besonderem Nutzen in epidemiologischen Feldstudien, bei denen die Kultivierung der Erreger besonders in Entwicklungsländern extrem schwierig sein kann. / This study was carried out in clusters of villages that represent an endemic focus of visceral leishmaniasis (VL). These villages were located in Gedaref state, eastern Sudan. For diagnostic purposes polymerase chain reaction (PCR) was performed successfully, directly from clinical samples spotted on filter papers with no prior cultivation from 100 patients suspected of having kala-azar or post kala-azar dermal leishmaniasis. Mainly the ribosomal internal transcribed spacer (ITS1 & ITS2) were targeted in PCR because this region is more variable and allows clear species identification and also strain differences could be expected by further analysis of these PCR products. PCR was found to be more sensitive compared to the gold standard microscopic method. Four PCR based approaches were used to analyse diversity within Sudanese isolates of Leishmania donovani. Methods compared were fingerprinting with single non-specific primers, restriction analysis of the amplified ITS locus (RFLP), single-stranded conformation polymorphism (SSCP) of the ITS region, major surface protease (gp63) gene, anonymous DNA fragments and sequencing of these targeted regions. When PCR fingerprinting and restriction analysis of ITS region were applied, highly similar fragment patterns were observed for all strains of L. donovani studied. The ITS1 locus gave 12 different SSCP profiles among the 86 Sudanese isolates, where as the ITS2 locus was highly conserved among the 86 samples with the exception of 1 isolate. Strains of L. donovani derived from other geographical areas were found to have different ITS2 patterns. The gp63 locus gave 3 polymorphic patterns among 31 Sudanese isolates. Concerning most of the anonymous DNA fragments namely, L510, L413, LK413, L0308 and L0114 unfortunately, we succeeded to get good PCR products only from DNA extracted 8 successful cultures. Only for the fragment L0110 we were able to get good PCR products from 31 samples spotted on filter papers. When these PCR products were investigated for polymorphisms using SSCP no differences were observed with exception of L0114 region, which showed 2 patterns. SSCP analysis correlates well with results of DNA sequencing and confirmed that SSCP was able to detect genetic diversity at the level of a single nucleotide. SSCP had advantages over the other methods employed for investigating of sequence variation within the species L. donovani. There was no correlation between the form of clinical manifestation of the disease and the PCR fingerprinting, ITS-RFLP, or ITS-SSCP characteristics. This study is beneficial particularly in epidemiological studies based on field-work where obtaining cultures can be extremely difficult especially in developing countries.
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"Avaliação da reação em cadeia de polimerase (PCR) no diagnóstico da leishmaniose cutânea no Estado do Espírito Santo, Brasil" / Evaluation of polymerase chain reaction (PCR) in the diagnosis of cutaneous leishmaniosis in the Espírito Santo state, BrazilLuciana Neves Passos 09 October 2003 (has links)
A identificação de espécie de Leishmania é crucial no diagnóstico de leishmaniose cutânea (LC), para terapia e prognóstico, em áreas com diferentes espécies endêmicas. A reação em cadeia de polimerase (PCR) foi usada em amostras de biópsias estocadas em parafina e formol de pacientes com LC, do Espírito Santo, Brasil. Usando sequências de mini-exon do gênero Leishmania, 63,2% (36/57) das amostras em parafina e 46,1% (6/13) das amostras em formol foram positivas. Numa abordagem usando sequências de kDNA e RFLP, 100% (58/58) das amostras de parafina testadas foram identificadas como Leishmania (V.) braziliensis. Estas reações podem ser usadas tanto para diagnóstico como para definição da espécie infectante / The identification of Leishmania species, a crucial step in cutaneous leishmaniasis, had therapeutics and prognostics implications, specially at when several agent species are endemic. Polymerase chain reaction (PCR) was used for analysis of paraffin-embedded and formalin stored skin biopsies from patients with parasitologic confirmed cutaneous formalin stored skin biopsies from patients with parasitologic confirmed cutaneous leishmaniasis, from the Espirito Santo State, Brazil. Tested by PCR targeted to mini-exon genus specific primer, 63,2% (36/57) of parafin and 46,1% (6/13) of formalin stored samples were positive, but using a PCR for kDNA primer followed by RFLP analysis, only Leishamnia (V.) braziliensis was identified in 100% (58/58) of parafin biopsies studied. Those reactions allow either diagnosis or species identification in cutaneous leismaniasis
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Caractérisation par microspectroscopie confocale Raman de la diffusion cutanée d'actif : optimisation des paramètres instrumentaux et méthodologiques en vue d'applications in vivo. / Characterization of actives diffusion through the skin by confocal Raman microspectroscopy : optimization of instrumental and methodological parameters for in vivo applications.Tfaili, Sana 14 February 2012 (has links)
La microspectroscopie Raman confocale apparait comme un outil à fort potentiel pour l'analyse in vivo de la peau, avec des applications innovantes en dermatologie et cosmétologie. Cette technique biophotonique permet d'accéder à des informations moléculaires très spécifiques d'un échantillon ; et ceci de façon totalement non destructive et sans aucun marquage ni préparation particulière. Les travaux référencés ont porté sur la caractérisation des constituants cutanés, l'évaluation du taux d'hydratation, le suivi de la perméation d'actif ou encore le diagnostic des lésions tumorales de la peau. Ces études ont été réalisées avec des configurations instrumentales très diverses. En vue de la conception d'une nouvelle micro-sonde Raman confocale, cette thèse a permis d'établir un cahier des charges précis ; les paramètres instrumentaux ont été définis et leurs effets sur la qualité des enregistrements Raman (fluorescence parasite, rapport signal sur bruit, atténuation du signal en profondeur) ont été évalués. En particulier, l'effet de la longueur d'onde d'excitation, paramètre clé en spectroscopie Raman, a été analysé. Dans nos expérimentations, la répétabilité et la variabilité du signal Raman ont également été prises en compte. Dans le but d'évaluer l'approche Raman pour l'analyse de la perméation cutanée d'actif, la diffusion cutanée de deux molécules (caféine et resvératrol) appliquées en faibles concentrations a été suivie sur un microspectromètre Raman confocal avec des paramètres d'acquisition similaires à ceux définis pour la micro-sonde. Nous avons montré la possibilité d'enregistrer des cinétiques de diffusion, et également mis en évidence des points limitatifs spécifiques de ces études expérimentales. / Confocal Raman microspectroscopy seems to be a tool with strong potential for in vivo skin analyses, and for innovative applications in dermatology and cosmetology fields. This biophotonic technique allows access to very specific molecular information non-destructively and without any particular skin labeling or sample preparation. The referenced work focused on the characterization of the cutaneous constituents, the evaluation of the rate of skin hydration, the monitoring of active ingredient permeation or diagnosis of tumoral skin lesions. These studies were performed with a variety of instrumental configurations. In view of the design of a new confocal Raman micro-probe, this thesis has established a registry of precise specifications; the instrumental parameters were defined, and their effects on the quality of the Raman records (parasite fluorescence, signal to noise ratio, signal attenuation in depth) were evaluated. In particular, the impact of the excitation wavelength, a key parameter in Raman spectroscopy, was analyzed. In our experiments, the reproducibility and the variability of the Raman signal were also considered. In order to evaluate the Raman approach for the analysis of cutaneous permeation of active ingredients, the cutaneous diffusion of two molecules (caffeine and resveratrol) applied in low concentrations were monitored by confocal Raman microspectroscopy using the acquisition parameters defined for the micro-probe. We have demonstrated the possibility of recording diffusion kinetics and at the same time unraveled specific limitations of these experimental studies.
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Efeito do fator de crescimento insulina-símile-I em promastigota e amastigota intracelular de Leishmania (Viannia) braziliensis de pacientes com diferentes formas clínicas de leishmaniose tegumentar americana / Effect of Insulin-like growth factor-I on promastigotes and intracellular amastigotes of Leishmania (Viannia) braziliensis from patients with different clinical forms of American tegumentary leishmaniasisSouza, Luana Dias de 03 October 2012 (has links)
Leishmanioses são doenças causadas por protozoários do gênero Leishmania e se apresentam sob forma tegumentar ou visceral. No Brasil, a leishmaniose tegumentar americana (LTA) é causada, na sua maioria, por Leishmania (Viannia) braziliensis e conhecem-se principalmente as formas cutânea (LC), mucosa (LM) e disseminada (LD) da doença. Na LTA as formas clínicas tem sido atribuídas a diferenças na resposta imune do hospedeiro, mas recentemente vinculam-se também à variabilidade intraespecífica da L. (V.) braziliensis. Neste estudo avaliamos se haveria variabilidade biológica nos isolados de L. (V.) braziliensis, provenientes de pacientes com LC, LM e LD, principalmente em resposta a fator de crescimento insulina-símile-I (IGF-I). Os fatores de crescimento do hospedeiro tem sido alvo de estudos no desenvolvimento das leishmanioses, sendo IGF-I um deles. Havíamos demonstrado em estudos anteriores, utilizando Leishmania (Leishmania) amazonensis, que IGF-I induz proliferação, aumentando a atividade da arginase, com geração de poliaminas e diminuindo a síntese de óxido nítrico. No presente estudo analisamos o efeito de IGF-I em L.(V.) braziliensis, espécie prevalente no Brasil. Avaliamos inicialmente as características dos diferentes isolados enquanto promastigota e no prosseguimento enquanto amastigota em células de linhagem monocítica humana THP-1, com e sem estímulo de IGF-I. Nossos dados sugerem que há diferenças na atividade da arginase basal entre os isolados de L. (V.) braziliensis, sendo maior naqueles provenientes de pacientes com LM. IGF-I aumentou a atividade da arginase nos isolados de LC e LD, mas não de LM. Nos isolados em forma amastigota nas células de linhagem monocítica humana THP-1, o efeito de IGF-I foi de aumento do parasitismo nos isolados de LC e LM e de diminuição com os de LD. Nos isolados de LD a atividade da arginase basal foi menor que nos de LC. Por outro lado, a produção de óxido nítrico tendeu a ser maior em isolados de LD quando sob estímulo de IGF-I. Os dados sugerem que diferenças nas características biológicas dos parasitos podem contribuir na apresentação clínica dos casos da LTA. / Leishmaniasis are diseases caused by protozoa of the genus Leishmania that may manifest as cutaneous or visceral disease. In Brazil, American tegumentary leishmaniasis (ATL) is caused mostly by Leishmania (Viannia) braziliensis and cutaneous (CL), mucosal (ML) and disseminated (DL) forms of the disease are known.The diversity of clinical manifestations has been attributed to differences in the host immune response, but recently it has also been related to intraspecific variability of L. (V.) braziliensis. In the present study we evaluated whether there were biological variability in different isolates of L. (V.) braziliensis from patients with CL, ML, and DL, mainly in response to insulin-like growth factor-I (IGF-I). Growth factors of the host have been investigated in the development of leishmaniasis including IGF-I. In previous studies using Leishmania (Leishmania) amazonensis IGF-I was shown to induce proliferation, to increase the activity of arginase, generating polyamines and to decrease the synthesis of nitric oxide. In this study we analyzed the effect of IGF-I in L. (V.) braziliensis, a species prevalent in Brazil. Initially we evaluated the characteristics of individual isolates as promastigote and further as amastigote within human macrophage cell line THP-1 with and without IGF-I stimulation. Our data suggest that there are differences in the basal arginase activity amongst isolates of L. (V.) braziliensis, being higher in those from patients with ML. IGF-I increased the activity of arginase in the isolates of CL and DL, but not of ML. In isolates in the form of amastigotes within THP-1 cells, IGF-I induced the increase of parasitism of isolates from CL and ML, and decrease of those from DL. In isolates of DL the basal arginase activity was lower than in those of CL. Moreover, the production of nitric oxide tended to be higher with isolates of DL upon IGF-I stimulation. The data suggest that differences in the biological characteristics of parasites may contribute to the diversity of clinical presentation of ATL.
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