Objetivo: Verificar se existe concordância entre examinadores ao avaliar o músculo orbicular superior da boca por meio de palpação, em indivíduos com e sem fissura labiopalatina e verificar se existe correlação dos níveis de atividade eletromiográfica obtidos durante a protrusão labial e durante a emissão da sílaba /pa/, com o grau de tonicidade muscular atribuído pelos examinadores. Modelo: Análise prospectiva comparando um grupo de indivíduos com fissura labiopalatina reparada (GF) e indivíduos sem fissura labiopalatina (GC), a um nível de significância de 5%. Local de Execução: Laboratório de Fisiologia, HRAC-USP. Participantes: 34 indivíduos com fissura labiopalatina transforame unilateral reparada, e 35 indivíduos não fissurados, todos com idade entre 18 a 30 anos. Resultados: os níveis de concordância entre os examinadores ao avaliarem por meio de palpação o músculo orbicular superior são próximos de zero, tanto no GF (E1xE2=0,19; E1xE3= -0,04; E2xE3=0,09) quanto no GC (E1xE2=-0,18; E1xE3=-0,05; E2xE3=-0,07). A correlação entre níveis de atividade eletromiográfica e palpação muscular pelos examinadores por meio de palpação do músculo orbicular superior obtidos durante a protrusão labial (R=-0,002; p=0,983) e durante a emissão da sílaba /pa/ (R=0,120; p=0,325), é negativa. Conclusões: Não houve concordância entre os examinadores ao avaliarem, por meio de palpação, o músculo orbicular superior, tanto para os indivíduos do grupo controle (GC) quanto para os indivíduos do grupo experimental (GF); Houve correlação negativa dos níveis de atividade eletromiográfica obtidos durante a protrusão labial e durante a emissão da sílaba /pa/, com o grau de tonicidade muscular atribuído pelos examinadores. / Objective: To verify if there is concordance between examinators when analyzing the superior orbicularis oris through palpation in individuals with and without cleft lip and palate, and verify if there is correlation of the electromyographic activity levels obtained during lips under protrusion and during /pa/ syllable emission, with muscle degree tonicity ascribed by the examinators. Model: A prospective analysis comparing a group of individuals with cleft lip and surgically repaired palate (FG) and individuals without cleft lip and palate (CG), at a significance level of 5%. Setting: Laboratory of Physiology, HRAC-USP. Participants: 34 individuals with unilateral cleft lip and surgically repaired palate and 35 normal individuals, between the ages of 18 and 30 years old. Results: The concordance levels between examinators when analyzing superior orbicularis oris through palpation are close to zero, for GF (E1xE2=0,19; E1xE3=-0,04; E2xE3=0,09) and for GC (E1xE2=-0,18; E1xE3=-0,05; E2xE3=-0,07). The correlation between electromyographic activity level and muscular palpation of the superior orbicularis oris by the examinators, obtained during lips under protrusion (R=-0,002; p=0,983) and during /pa/ syllable emission (R=0,120; p=0,325), is negative. Conclusion: There was no concordance between the examinators when analyzing, through palpation, the superior orbicular oris for both individuals from control group (GC) and from the experimental group (GF). There was negative correlation of the electromyographic activity levels obtained during lips under protrusion and during /pa/ syllable emission, with muscle degree tonicity ascribed by the examinators.

Raktiniai žodžiai: laikysena, skoliozė, raumenų tonusas, kineziterapija. Tyrimo objektas: 10-16 metų (5,6,7,8 klasių) vaikų, sergančių cerebriniu paralyžiumi laikysenos sutrikimai ir korekcija. Tyrimo problema: Netaisyklinga laikysena, ydinga laikysena, skoliozė – vis dažniau skambančios frazės ugdymo įstaigose šiomis dienomis. Šiame kompiuterių ir technikos amžiuje laikysenos sutrikimai yra labai dažnas reiškinys mokinių tarpe. Vaikams, sergantiems cerebriniu paralyžiumi, ši problema yra ypač aktuali, nes jų kūno laikyseną labai įtakoja jų specifinis raumenų tonusas, sąnarių kontraktūros, jau nekalbant apie nepritaikytus mokyklinius suolus ir laiką, praleistą prie kompiuterio. Jų kūno laikyseną reikia nuolat prižiūrėti, intensyvinti mankštas, taikyti įvairius metodus, naujoves, kad pasiekti pačią optimaliausia formą. Darbo tikslas: Išsiaiškinti, ar pakito tiriamųjų laikysena bei raumenų jėga ir tonusas, pritaikius judesių korekcijos užsiėmimų programą. Tyrimo uždaviniai: 1. Apibendrinti literatūrą apie vaikų laikysenos sutrikimus, vaikų, sergančių cerebriniu paralyžiumi raumenų tonuso ypatumus. 2. Įvertinti cerebriniu paralyžiumi sergančių vaikų laikyseną, sąnarių amplitudes, raumenų tonusą bei jėgą prieš judesių korekcijos užsiėmimų programos sudarymą. 3. Ištyrus sudaryti judesių korekcijos užsiėmimų bendrą programą (bendrą ir jei reikia individualią); 4. Įvertinti cerebriniu paralyžiumi sergančių vaikų laikyseną, raumenų tonusą bei jėgą judesių korekcijos užsiėmimų... [toliau žr. visą tekstą] / Key words: posture, scoliosis, muscular tonicity, kinezitherapy. The object of research: posture disorder and correction of 10-16 year (5th, 6th, 7th and 8th forms) children having cerebral paralysis. The problem of research: Incorrect posture, perverse posture and scoliosis are phrases heard and pronounced more and more often at educational institutions nowadays. In the computer and machine age posture disorder is a very common phenomenon among schoolchildren. The problem is especially relevant for children having cerebral paralysis as their body posture is extremely influenced by specific muscular tonicity and articular contractor not to mention unfitted school desks and time spent in front of the computer. In order to achieve the most optimum physical form these children’s body posture must always be under observation, exercises should be intensified and various methods and innovations applied. The purpose of research: To ascertain if children’s in the study posture and muscular power and tonicity have changed since the application of the programme of activities of movement correction. The objectives of research: 1. To summarize literature on the disorder of children’s posture and peculiarities of muscular tonicity of children having cerebral paralysis. 2. To evaluate posture, articular amplitude, muscular tonicity and power of children having cerebral paralysis before formation of the programme of activities of movement correction. 3. To form a general programme of... [to full text]

Objetivo: Verificar se existe concordância entre examinadores ao avaliar o músculo orbicular superior da boca por meio de palpação, em indivíduos com e sem fissura labiopalatina e verificar se existe correlação dos níveis de atividade eletromiográfica obtidos durante a protrusão labial e durante a emissão da sílaba /pa/, com o grau de tonicidade muscular atribuído pelos examinadores. Modelo: Análise prospectiva comparando um grupo de indivíduos com fissura labiopalatina reparada (GF) e indivíduos sem fissura labiopalatina (GC), a um nível de significância de 5%. Local de Execução: Laboratório de Fisiologia, HRAC-USP. Participantes: 34 indivíduos com fissura labiopalatina transforame unilateral reparada, e 35 indivíduos não fissurados, todos com idade entre 18 a 30 anos. Resultados: os níveis de concordância entre os examinadores ao avaliarem por meio de palpação o músculo orbicular superior são próximos de zero, tanto no GF (E1xE2=0,19; E1xE3= -0,04; E2xE3=0,09) quanto no GC (E1xE2=-0,18; E1xE3=-0,05; E2xE3=-0,07). A correlação entre níveis de atividade eletromiográfica e palpação muscular pelos examinadores por meio de palpação do músculo orbicular superior obtidos durante a protrusão labial (R=-0,002; p=0,983) e durante a emissão da sílaba /pa/ (R=0,120; p=0,325), é negativa. Conclusões: Não houve concordância entre os examinadores ao avaliarem, por meio de palpação, o músculo orbicular superior, tanto para os indivíduos do grupo controle (GC) quanto para os indivíduos do grupo experimental (GF); Houve correlação negativa dos níveis de atividade eletromiográfica obtidos durante a protrusão labial e durante a emissão da sílaba /pa/, com o grau de tonicidade muscular atribuído pelos examinadores. / Objective: To verify if there is concordance between examinators when analyzing the superior orbicularis oris through palpation in individuals with and without cleft lip and palate, and verify if there is correlation of the electromyographic activity levels obtained during lips under protrusion and during /pa/ syllable emission, with muscle degree tonicity ascribed by the examinators. Model: A prospective analysis comparing a group of individuals with cleft lip and surgically repaired palate (FG) and individuals without cleft lip and palate (CG), at a significance level of 5%. Setting: Laboratory of Physiology, HRAC-USP. Participants: 34 individuals with unilateral cleft lip and surgically repaired palate and 35 normal individuals, between the ages of 18 and 30 years old. Results: The concordance levels between examinators when analyzing superior orbicularis oris through palpation are close to zero, for GF (E1xE2=0,19; E1xE3=-0,04; E2xE3=0,09) and for GC (E1xE2=-0,18; E1xE3=-0,05; E2xE3=-0,07). The correlation between electromyographic activity level and muscular palpation of the superior orbicularis oris by the examinators, obtained during lips under protrusion (R=-0,002; p=0,983) and during /pa/ syllable emission (R=0,120; p=0,325), is negative. Conclusion: There was no concordance between the examinators when analyzing, through palpation, the superior orbicular oris for both individuals from control group (GC) and from the experimental group (GF). There was negative correlation of the electromyographic activity levels obtained during lips under protrusion and during /pa/ syllable emission, with muscle degree tonicity ascribed by the examinators.

While machine perfusion of explanted kidneys is theoretically superior to standard cold storage, it may damage potential transplants unless machine-associated swelling is controlled. This thesis presents the effects of perfusate tonicity on renal swelling during hypothermic machine perfusion. Phosphate buffered solution (PBS) and PBS supplemented with 5% w/v mannitol were used as isotonic (289 mOsm/kg) and hypertonic (568 mOsm/kg) perfusates, respectively. Porcine kidney pairs were procured then flushed and machine perfused; the right and left kidneys were assigned opposite perfusates. An experimental methodology was developed to image porcine kidneys undergoing hypothermic machine perfusion (5 deg C) for 15 minutes followed by 120 minutes without perfusion to quantify surface displacement (renal swelling) with digital image correlation (DIC). Surface displacement and size (thickness) were compared between the right and left kidneys of each pair. In addition, discharged renal fluids (i.e., filtrate and venous outflow) and biopsies were collected. On average, kidneys perfused with the mannitol solution were smaller in size than the kidneys perfused with PBS (p < 0.05) at the start and end of each experiment; however, there was no significant difference between the renal sizes at the end of the 15 minute perfusion interval (p > 0.05). Thus, hypertonic and isotonic perfusates yielded different renal swelling outcomes (i.e., physical size and surface displacement), which suggests that perfusate tonicity influences renal swelling. These experiments are the first time ex vivo renal surface displacement measurements have been collected during machine perfusion. / Master of Science

Cytochrome P450 3A isozymes (CYP3A) metabolize approximately 50% of therapeutic drugs. It has recently been discovered that human CYP3A mRNA levels can be induced by hypertonicity; a physiological state not previously linked to its regulation. The osmosensitive transcription factor, Nuclear Factor of Activated T-Cells 5 (NFAT5), regulates multiple genes that restore osmolyte homeostasis and promote cell protection during osmotic stress. In silico examinations and in vitro experiments using reporters, knockdown and binding assays in the human intestinal cell line C2bbe1 have revealed an active tonicity-responsive enhancer (TonE) within CYP3A7 intron (+5417/+5427 from CYP3A7 transcriptional start site) that is responsible for NFAT5 binding and NFAT5-dependent regulation of CYP3A isoforms. In addition, hypertonicity-mediated CYP3A induction is also observed in both hepatic and intestinal cell lines. Effects of tonicity changes on in vivo CYP3A expression and function were examined in a humanized CYP3A transgenic mouse with similar tissue expression in humans. More specifically, intervention with prolonged dehydration involving alternating between 24-hour cycles of water-deprivation and water ad lib for 1 week (cyclic water-deprivation; four 24-hour water-deprivation and three 24-hour water ad lib periods), increased expression of NFAT5 target genes Slc6a12 in the liver and kidney (2.5 ± 0.6-fold over water ad lib, n = 14, p = 0.04; and 3.1 ± 0.6-fold, n = 10, p = 0.02, respectively), Akr1b3 in the liver, and Slc5a3 in the kidney. Immunofluorescent microscopy revealed an increase of nuclear-distributed mouse NFAT5 in cyclic water-deprived animals, consistent with NFAT5 activation. Most importantly, CYP3A4 mRNA levels were noted to be elevated in the liver and kidney (11.8 ± 4.8-fold over water ad lib, n = 14, p = 0.04 and 2.2 ± 0.4-fold, n = 9, p = 0.02, respectively), with concurrent CYP3A protein and activity increase. Localized hypertonic environment in the gut was simulated by providing animals with a week-long high-salt diet. The effects of high-salt diet in the gut were similar to those of cyclic water-deprivation in the liver and kidney; where NFAT5 showed nuclear distribution and NFAT5 target gene expression (Slc6a12; 20.5 ± 6.7-fold over a week-long low-salt diet, n = 8, p = 0.02 and Slc6a6; 3.2 ± 0.7-fold, n = 10, p < 0.01, in the duodenum). Furthermore, an increase of CYP3A4 mRNA was observed (2.6 ± 0.5-fold over a week-long low-salt diet, n = 14, p = 0.03), with a corresponding rise in protein expression and activity levels. In summary, increased expression of in vitro and in vivo human CYP3A was achieved using a hypertonic stimulus; concurrent NFAT5 activation and NFAT5 target gene expression were observed. These results suggested a possible binding of activated NFAT5 to CYP3A TonE situated within the intronic region of CYP3A7. It could be further concluded that NFAT5 may be responsible for the hypertonic induction of human CYP3A.

Cytochrome P450 3A isozymes (CYP3A) metabolize approximately 50% of therapeutic drugs. It has recently been discovered that human CYP3A mRNA levels can be induced by hypertonicity; a physiological state not previously linked to its regulation. The osmosensitive transcription factor, Nuclear Factor of Activated T-Cells 5 (NFAT5), regulates multiple genes that restore osmolyte homeostasis and promote cell protection during osmotic stress. In silico examinations and in vitro experiments using reporters, knockdown and binding assays in the human intestinal cell line C2bbe1 have revealed an active tonicity-responsive enhancer (TonE) within CYP3A7 intron (+5417/+5427 from CYP3A7 transcriptional start site) that is responsible for NFAT5 binding and NFAT5-dependent regulation of CYP3A isoforms. In addition, hypertonicity-mediated CYP3A induction is also observed in both hepatic and intestinal cell lines. Effects of tonicity changes on in vivo CYP3A expression and function were examined in a humanized CYP3A transgenic mouse with similar tissue expression in humans. More specifically, intervention with prolonged dehydration involving alternating between 24-hour cycles of water-deprivation and water ad lib for 1 week (cyclic water-deprivation; four 24-hour water-deprivation and three 24-hour water ad lib periods), increased expression of NFAT5 target genes Slc6a12 in the liver and kidney (2.5 ± 0.6-fold over water ad lib, n = 14, p = 0.04; and 3.1 ± 0.6-fold, n = 10, p = 0.02, respectively), Akr1b3 in the liver, and Slc5a3 in the kidney. Immunofluorescent microscopy revealed an increase of nuclear-distributed mouse NFAT5 in cyclic water-deprived animals, consistent with NFAT5 activation. Most importantly, CYP3A4 mRNA levels were noted to be elevated in the liver and kidney (11.8 ± 4.8-fold over water ad lib, n = 14, p = 0.04 and 2.2 ± 0.4-fold, n = 9, p = 0.02, respectively), with concurrent CYP3A protein and activity increase. Localized hypertonic environment in the gut was simulated by providing animals with a week-long high-salt diet. The effects of high-salt diet in the gut were similar to those of cyclic water-deprivation in the liver and kidney; where NFAT5 showed nuclear distribution and NFAT5 target gene expression (Slc6a12; 20.5 ± 6.7-fold over a week-long low-salt diet, n = 8, p = 0.02 and Slc6a6; 3.2 ± 0.7-fold, n = 10, p < 0.01, in the duodenum). Furthermore, an increase of CYP3A4 mRNA was observed (2.6 ± 0.5-fold over a week-long low-salt diet, n = 14, p = 0.03), with a corresponding rise in protein expression and activity levels. In summary, increased expression of in vitro and in vivo human CYP3A was achieved using a hypertonic stimulus; concurrent NFAT5 activation and NFAT5 target gene expression were observed. These results suggested a possible binding of activated NFAT5 to CYP3A TonE situated within the intronic region of CYP3A7. It could be further concluded that NFAT5 may be responsible for the hypertonic induction of human CYP3A.

L’aldostérone et le Récepteur Minéralocorticoïde (MR) participent au contrôle de la balance hydrosodée et de la pression artérielle. Les altérations de l’expression du MR ou de la signalisation minéralocorticoïde sont associées à de nombreuses pathologies chez l’Homme. Dans ce travail, nous avons démontré, le rôle majeur de protéines de liaison à l’ARN, Tis11b et HuR, dans le contrôle post-transcriptionnel de l’expression du MR en réponse aux variations de tonicité extracellulaire dans un modèle de cellules principales rénales et chez la souris. L’hypertonicité (500 mOsmol/L) induit l’expression de la protéine Tis11b, qui lie la région 3’-non traduite du transcrit MR afin d’accélérer sa dégradation, diminuant ainsi l’expression rénale de la protéine MR et de la signalisation minéralocorticoïde. A l’opposé, l’hypotonicité (150 mOsmol/L) stimule la translocation nucléo-cytoplasmique de HuR, qui stabilise le transcrit MR, augmentant ainsi l’expression du MR et la sensibilité rénale à l’aldostérone. De plus, HuR est responsable de l’édition d’un nouveau variant d’épissage du MR, le variant MR Δ6, obtenu par l’exclusion de l’exon 6.Ce variant d’épissage exerce un effet dominant négatif sur la signalisation minéralocorticoïde. Enfin, l’identification de microARN modulés par l’hypertonicité suggère leur rôle potentiel dans le contrôle de la signalisation minéralocorticoïde rénale. La caractérisation de ces mécanismes inédits modulant l’action du MR améliore notre compréhension de la physiopathologie de la signalisation minéralocorticoïde, et pourrait aboutir, à terme, à de nouvelles stratégies thérapeutiques. / Aldosterone and the Mineralocorticoid Receptor (MR) participate to the control of salt and water balance and the arterial pressure. Alteration of renal MR expression or mineralocorticoid signaling pathway contributes to the development of numerous human disorders. In this work, we have demonstrated the major role played by the RNA-Binding Proteins, Tis11b and HuR, in the control of MR expression in response to variations of extracellular tonicity in a model of principal tubular cells and in vivo. Hypertonicity (500 mOsmol/L) increases the expression ofTis11b, which binds the 3’-untranslated region of MR transcript and accelerates the degradation of MR transcript, leading to the reduction of the mineralocorticoid signaling. Conversely, hypotonicity (150 mOsmol/L) stimulates nuclear-cytoplasmic shuttling of HuR protein, which stabilizes MR transcript increasing its expression and renal sensitivity to aldosterone action. Furthermore, HuR participates to the editing of the novel MR Δ6 splice variant, which lacks exon 6, and exerts a dominant negative effect on mineralocorticoid signaling. Finally, we have provided evidence that hypertonicity modulates expression of microRNA, which may control mineralocorticoid signaling pathway. Characterization of these original mechanisms modulating MR action is pivotal for a better understanding of mineralocorticoid-related pathophysiology, and should ultimately lead to the development of new therapeutic strategies.