Submitted by PPG Odontologia (odontologia-pg@pucrs.br) on 2018-03-26T12:53:16Z No. of bitstreams: 1 LEONARDO_MATOS_SANTOLIM_ZANETTINI_DIS.pdf: 2920184 bytes, checksum: f65b15c1056e1a9173b56c1789377ba8 (MD5) / Approved for entry into archive by Tatiana Lopes (tatiana.lopes@pucrs.br) on 2018-04-06T16:34:38Z (GMT) No. of bitstreams: 1 LEONARDO_MATOS_SANTOLIM_ZANETTINI_DIS.pdf: 2920184 bytes, checksum: f65b15c1056e1a9173b56c1789377ba8 (MD5) / Made available in DSpace on 2018-05-18T14:04:05Z (GMT). No. of bitstreams: 1 LEONARDO_MATOS_SANTOLIM_ZANETTINI_DIS.pdf: 2920184 bytes, checksum: f65b15c1056e1a9173b56c1789377ba8 (MD5) Previous issue date: 2018-01-10 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / . / Recombinant Human Growth Hormone (RhGh) has been studied in cases of dental implants, TMJ pathologies and bone fractures. It is able to stimulate bone growth in a dose-dependent manner by direct stimulation of chondrocytes. It stimulates proliferation and differentiation of chondroprogenitor cells, also acts directly on osteoblasts, increasing rates of remodeling and bone formation. In this review of the literature and case report, RhGh will be used in alveolar ridge preservation procedures, with the purpose of evaluating the benefits of this technique in maintaining the postextraction bone site viable, aiming a future rehabilitation with dental implants and prostheses.

Growth hormone-releasing hormone (GRF) and pituitary adenylate cyclase-activating polypeptide (PACAP) are members of the PACAP/Glucagon superfamily. The family is proposed to have developed from an ancestral PACAP-like molecule in invertebrates. Through successive exon, gene and genome duplications the family has grown to include seven other members. In mammals GRF and PACAP are located on different genes, but in fish, amphibians and birds they are located on the same gene. The main function of GRF is the release of growth hormone (GH) from the pituitary. Also, during development GRF influences the fetal pituitary and stimulates GH release during late gestation. In contrast, the functions of PACAP are extremely varied. PACAP is the newest member of the superfamily and there is still much work to be done before its actions are well understood. Like GRF, PACAP is a releasing hormone acting on the pituitary and in addition, the adrenal gland, pancreas and heart, as well as other organs. Also, PACAP regulates smooth muscle in the vascular system, gut, respiratory tract and reproductive tract During development PACAP affects proliferation, differentiation and apoptosis. GRF and PACAP are expressed throughout development in fish, beginning during the blastula period in rainbow trout and at the end of gastrulation in zebrafish (earliest stage examined). In rainbow trout the grf/pacap gene is expressed as two transcripts, a short and a long transcript. The short transcript is produced by alternative splicing of the gene and does not include the fourth exon which codes for GRF. The long transcript includes the coding regions for both GRF and PACAP. By this means PACAP can be regulated separately from GRF. With the extensive role PACAP appears to play in development, separate regulation of the hormone may be necessary. Expression of the grf/pacap gene in zebrafish is widespread early in development and gradually becomes localized. Of particular interest is the expression of the grf/pacap transcript in regions associated with the prechordal plate, an important organizing center in development. Although it is not yet confirmed, there is evidence to suggest GRF and PACAP are expressed in the prechordal plate and its derivatives in the gut and hatching gland. In addition, expression of the grf/pacap transcript is observed in the neuroectoderm (eye, brain and spinal cord) and the developing heart. Considering the expression pattern of GRF and PACAP, I propose that one of both of these hormones may be involved in patterning during vertebrate embryogenesis. The evolution of gene families is thought to occur through successive exon, gene and genome duplications. Duplicate exons or genes become differentiated and eventually gain new functions or become functionless. During evolution of the grf/pacap lineage, several duplication events have occurred. Analysis of rainbow trout leads me to think that this fish and other salmonids possess two copies of the grf/pacap gene. This is not unexpected considering the tetraploid nature of salmonids. Present day mammals encode GRF and PACAP on separate genes. At some point during the evolution of this lineage a duplication event has occurred, possibly in early mammals or prior to the divergence of birds. The study of multigene families is a useful way to understand evolutionary processes. To this end I examined three members of multigene families from sockeye salmon. Therefore, in addition to the evolutionary mechanisms and pathways that directed grf/pacap gene evolution, I examined the ferritin-H subunit, the alpha-tubulin subunit and the beta-globin subunit. These cDNA sequences are similar to their counterparts in other teleost. The evolution of the ferritin gene family is particularly interesting because it involves the addition or deletion of DNA sequences that affect regulation and cytosolic location. / Graduate

HMb, metabólito da leucina, utilizado para aumentar a síntese protéica. Investigamos o efeito do HMb sobre o eixo somatotrófico, bem como o mRNA de IGF-I e miostatina muscular. Ratos tratados com HMb (320 mg/Kg de peso corporal /mL de salina-0,9%), ou salina (controle), gavagem, 4 semanas, decapitados, sangue para avaliação sérica: insulina (RIE), glicose (colorimetria) e IGF-I (RIE). Extração de RNA total, para avaliação do mRNA de IGF-I e miostatina (Fígado, músculo extensor digital longo, Sóleo), avaliação da expressão do mRNA do GH, por Northern Blot, e expressão do GH ,Western blotting (hipófise). Dados analisados pelo teste-T de Student (P<0,05). Tratamento aumentou o conteúdo de mRNA de GH (> 60%), da proteína GH (>20%), do mRNA do IGF-I (~24%), da concentração sérica de IGF-I (p<0,05), indicando uma ativação do eixo somatotrófico pelo HMb, sem alterações no mRNA de miostatina e IGF-I muscular, ainda um aumento da insulina (~2x), sem alterações na glicose sérica, resultado do efeito hiperglicemiante do GH, ou um efeito direto do HMb na secreção de insulina. / HMb, metabolite of leucine, used to increase protein synthesis. Evaluate the effect HMb on the somatotrophic axis activity, as well as muscle mRNA IGF-I and myostatin. Rats treated with HMb (320 mg / kg body weight / mL of saline-0, 9%) or saline (control), gavage, 4 weeks, decapitated, blood for evaluation of serum: insulin (RIA), glucose (colorimetric) and IGF-I (RIA). Extraction of RNA total, for evaluation the mRNA IGF-I and myostatin (liver, muscle extensor digitalis longus (EDL) and soleus), evaluation of the GH mRNA expression of by Northern blot, and GH content, western blotting (pituitaries). Data analyzed by Student t-test (P <0.05). HMb treatment increased the content of GH mRNA (> 60%), GH (> 20%), IGF-I mRNA (~ 24%), IGF-I (p <0.05), indicates that the somatotrophic axis activity is increased by the HMb, without changes in mRNA of myostatin and muscle IGF-I, insulin also increased (~ 2x), without changes in serum glucose, hyperglycemiant result of the effect of GH or a direct effect of HMb in the secretion of insulin.

Made available in DSpace on 2014-06-11T19:27:18Z (GMT). No. of bitstreams: 0 Previous issue date: 2011-02-07Bitstream added on 2014-06-13T18:31:17Z : No. of bitstreams: 1 batista_eos_me_araca.pdf: 385042 bytes, checksum: a09da05bf31ae74120f8fbc231868087 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Foi avaliada a variação na concentração de GH em resposta a agonistas ou antagonistas de neurotransmissores durante o período pré-púbere em 10 novilhas da raça Nelore. A administração de um antagonista gabaérgico (picrotoxina, 0,18 mg/kg, I.V., amostras a cada 15 min por 10 h) aumentou a concentração de GH aos 10 meses de idade, mas não aos 14 e 17 meses. Tanto no grupo Picrotoxina, como no Controle a concentração média de GH foi maior aos 10 meses do que aos 17 meses de idade. O grupo picrotoxina apresentou aos 17 meses maior área de concentração de GH que o controle. O grupo picrotoxina apresentou aos 14 meses maior área total de picos que o controle. Aos 10 e 14 meses no grupo Picrotoxina houve maior amplitude de picos do que aos 17 meses. Bloqueando o receptor (D2) dopaminérgico (sulpiride, 0,59 mg/kg, SC, amostras a cada 15 min por 10 h) houve aumentou na concentração de GH aos 8 e 12 meses, mas não aos 16 meses de idade, comparado ao grupo controle. No grupo Sulpiride a concentração média de GH foi maior aos 8 e 12, comparado com os 16 meses de idade. Em relação à área total de secreção, área total de picos, e amplitude máxima de picos de secreção de GH não houve diferença entre os grupos. Aos 8 meses o grupo Sulpiride apresentou menor tempo, porém aos 12 meses o tempo foi maior para o primeiro pico em relação ao controle. O tempo para o primeiro pico foi menor aos 8 e 16 meses que aos 12 meses no grupo Sulpiride. No grupo controle aos 12 meses o tempo para o primeiro pico foi menor que aos 8 e 16 meses. Aos 16 meses o grupo Sulpiride apresentou maior freqüência de picos que o Controle. A aplicação de um estimulador alfa-adrenérgico (clonidina, 10 µg/kg, I.V., amostras a cada 15 min por 4 h) aumentou a concentração média de GH aos 8 e 15 meses em relação ao grupo Controle e em alguns intervalos aos 12 meses... / This study evaluated the GH concentration after neurotransmitter agonist or antagonists injection in prepubertal Nelore heifers at different ages. A gabaergic antagonist (picrotoxin, 0.18 mg/kg,I.V., samples every 15 min for 10h) promoted an increase on GH concentration at 10 months of age but not at 14 and 17 mo. In both Picrotoxin and Control groups GH average concentration was higher at 10 mo than at 17 month of age. At 17 mo the picrotoxin group had a larger GH concentration area than control. There was no difference among the groups on the time for the first GH surge, but at 14 mo the Picrotoxin group had a higher total peak area than control. At 10 and 14 mo the Picrotoxin group had an higher peak amplitude than at 17 months. Blocking the D2-dopaminergic receptor (Sulpiride 0.59 mg/kg, SC, samples every 15 min for 10 h) there was an increase on GH secretion at 8 and 12 months, but not at 16 mo of age, compared to control group. Inside Sulpiride group the average GH concentration was higher at 8 and 12 mo than at 16 month of age. For both total area, peaks total area, time for first peak, and peak maximal amplitude there was no difference among groups. The time for the first GH peak was lower for Sulpiride compared to Control group at 8 mo, but at 12 mo the time was higher than in Control. Inside Sulpiride group time was lower at 8 and 16 month compared to 12. In the control group time for the first peak was lower at 12 than 8 and 16 months. At 16 month the Sulpiride group had higher peak frequency than Control. An alpha adrenergic stimulation (Clonidine, 10 µg/kg, IV, samples every 15 min for 4 h) increased mean GH concentration at 8 and 15 months compared to Control group and at some intervals at 12 months. At 8 months Clonidine group had higher total secretion area, total peak area, maximum peak amplitude and lowe time for the first GH secretion peak, compared... (Complete abstract click electronic access below)

Orientador: Guilherme de Paula Nogueira / Banca: Cláudia Maria Bertan Membrive / Banca: Ciro Moraes Barros / Resumo: Foi avaliada a variação na concentração de GH em resposta a agonistas ou antagonistas de neurotransmissores durante o período pré-púbere em 10 novilhas da raça Nelore. A administração de um antagonista gabaérgico (picrotoxina, 0,18 mg/kg, I.V., amostras a cada 15 min por 10 h) aumentou a concentração de GH aos 10 meses de idade, mas não aos 14 e 17 meses. Tanto no grupo Picrotoxina, como no Controle a concentração média de GH foi maior aos 10 meses do que aos 17 meses de idade. O grupo picrotoxina apresentou aos 17 meses maior área de concentração de GH que o controle. O grupo picrotoxina apresentou aos 14 meses maior área total de picos que o controle. Aos 10 e 14 meses no grupo Picrotoxina houve maior amplitude de picos do que aos 17 meses. Bloqueando o receptor (D2) dopaminérgico (sulpiride, 0,59 mg/kg, SC, amostras a cada 15 min por 10 h) houve aumentou na concentração de GH aos 8 e 12 meses, mas não aos 16 meses de idade, comparado ao grupo controle. No grupo Sulpiride a concentração média de GH foi maior aos 8 e 12, comparado com os 16 meses de idade. Em relação à área total de secreção, área total de picos, e amplitude máxima de picos de secreção de GH não houve diferença entre os grupos. Aos 8 meses o grupo Sulpiride apresentou menor tempo, porém aos 12 meses o tempo foi maior para o primeiro pico em relação ao controle. O tempo para o primeiro pico foi menor aos 8 e 16 meses que aos 12 meses no grupo Sulpiride. No grupo controle aos 12 meses o tempo para o primeiro pico foi menor que aos 8 e 16 meses. Aos 16 meses o grupo Sulpiride apresentou maior freqüência de picos que o Controle. A aplicação de um estimulador alfa-adrenérgico (clonidina, 10 µg/kg, I.V., amostras a cada 15 min por 4 h) aumentou a concentração média de GH aos 8 e 15 meses em relação ao grupo Controle e em alguns intervalos aos 12 meses... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: This study evaluated the GH concentration after neurotransmitter agonist or antagonists injection in prepubertal Nelore heifers at different ages. A gabaergic antagonist (picrotoxin, 0.18 mg/kg,I.V., samples every 15 min for 10h) promoted an increase on GH concentration at 10 months of age but not at 14 and 17 mo. In both Picrotoxin and Control groups GH average concentration was higher at 10 mo than at 17 month of age. At 17 mo the picrotoxin group had a larger GH concentration area than control. There was no difference among the groups on the time for the first GH surge, but at 14 mo the Picrotoxin group had a higher total peak area than control. At 10 and 14 mo the Picrotoxin group had an higher peak amplitude than at 17 months. Blocking the D2-dopaminergic receptor (Sulpiride 0.59 mg/kg, SC, samples every 15 min for 10 h) there was an increase on GH secretion at 8 and 12 months, but not at 16 mo of age, compared to control group. Inside Sulpiride group the average GH concentration was higher at 8 and 12 mo than at 16 month of age. For both total area, peaks total area, time for first peak, and peak maximal amplitude there was no difference among groups. The time for the first GH peak was lower for Sulpiride compared to Control group at 8 mo, but at 12 mo the time was higher than in Control. Inside Sulpiride group time was lower at 8 and 16 month compared to 12. In the control group time for the first peak was lower at 12 than 8 and 16 months. At 16 month the Sulpiride group had higher peak frequency than Control. An alpha adrenergic stimulation (Clonidine, 10 µg/kg, IV, samples every 15 min for 4 h) increased mean GH concentration at 8 and 15 months compared to Control group and at some intervals at 12 months. At 8 months Clonidine group had higher total secretion area, total peak area, maximum peak amplitude and lowe time for the first GH secretion peak, compared... (Complete abstract click electronic access below) / Mestre

nÃo hà / IntroduÃÃo: MutaÃÃes nos genes GNAS e AIP estÃo presentes em 35% e 3%, respectivamente, dos somatotrofinomas esporÃdicos. Recentemente, observa-se importÃncia biolÃgica crescente dos microRNAs (miRNAs) na tumorigÃnese hipofisÃria. Entretanto, mecanismos moleculares envolvidos na patogÃnese de 60% desses tumores permanecem nÃo elucidados. Objetivos: Identificar a prevalÃncia de mutaÃÃes nos genes GNAS e AIP em um grupo de somatotrofinomas esporÃdicos. Comparar parÃmetros clÃnicos e bioquÃmicos ao diagnÃstico como idade, tamanho tumoral e agressividade (escore Hardy), hormÃnio do crescimento (GH), prolactina (PRL) e Fator de Crescimento Insulin-Like I (IGF-1) e resposta as terapÃuticas entre os grupos com (gsp+) e sem (gsp-) mutaÃÃo no GNAS. Analisar a expressÃo dos genes GNAS e BTG2 e miRNAs entre somatotrofinomas e hipÃfises normais (HN) e a associaÃÃo entre a expressÃo com agressividade, a resposta à cirurgia e a todas as terapÃuticas adjuvantes disponÃveis. Material e MÃtodos: 26 pacientes com diagnÃstico de acromegalia. Tamanho tumoral foi avaliado por RNM/CT e o grau de invasibilidade pelo escore de Hardy (I a IV). GH basal &#8804;2.5&#956;g/L ou nadir de GH apÃs o GTT&#8804;1&#956;g/L e IGF-1 normal para idade e sexo foram utilizados como critÃrio de cura apÃs cirurgia transesfenoidal (CTE). Como controle com o anÃlogo da somatostatina (AS), adotamos a normalizaÃÃo dos nÃveis de IGF-1 para idade e sexo. As amostras tumorais (n=26) foram obtidas durante a CTE, realizado histopatolÃgico e armazenadas a -70 ÂC, para estudos moleculares. HN (07) foram obtidas durante autÃpsias. RNA e DNA total foram extraÃdos pelo TRIzolÂ. Os cÃdons 201 e 227 do gene GNAS e o AIP completo foram sequenciados. ExpressÃo relativa dos genes GNAS e BTG2 e dos miRNAs let-7a, miR-16a, miR-21, miR-141, miR-143, miR-15a, miR-145, miR-23a, miR-23b e miR-24-2 foi avaliada por qPCR (sondas TaqMan), pelo mÃtodo 2-&#916;&#916;Ct. Resultados: A frequÃncia de mutaÃÃes no GNAS foi de 35% e no AIP 3,8%. NÃo houve diferenÃa entre as mÃdias de idade (39,0Â11,5 vs 43,6Â9,0 anos; p=0,32), nas concentraÃÃes plasmÃticas basais de GH (62,4Â128,1 vs 39,9Â48,3Âg/L; p=0,39), IGF-1 (435,5Â230,8 vs 556,9 238,3 %ULNR; p=0,32), PRL (25,7Â29,8 vs 30,9Â32,8 ng/L; p=0,69) e agressividade tumoral entre os gsp+ e gsp-(p=1,00). Ao analisar o uso do AS como terapÃutica adjuvante à CTE, observamos que 04/05 (80%) dos indivÃduos com somatotrofinoma gsp+ obtiveram controle da doenÃa, enquanto que no grupo gsp- 02/06 (33%) obtiveram controle (p=0,08). Quando associamos ao AS, os agonistas dopaminÃrgicos e/ou radioterapia externa, observamos que 05/05 (100%) dos pacientes gsp+ tiveram critÃrio de controle da doenÃa, contra (04/09) 44% no grupo gsp- (p=0,09). NÃo houve diferenÃa na expressÃo de GNAS entre os somatotrofinomas e as HN (1,07Â0,55 vs 0,98Â0,28; p=0,97), e entre os gsp+ e gsp- (1,04Â0,59 vs 1,10Â0,55; p=0,97, respectivamente). Os tumores Hardy I / II apresentaram maior expressÃo do GNAS do que os tumores classificados como III / IV (p=0,02). NÃo houve associaÃÃo entre a expressÃo do GNAS e o controle da doenÃa com cirurgia isolada ou com o uso de todas as terapÃuticas adjuvantes. Observamos hipoexpressÃo do BTG2 e dos miR-16a e miR-141 em somatotrofinomas quando foram comparados com as HN (p=0,002, fold=-6,63; p=0,01, fold=-10,00; p=0,0003, fold=-50,00, respectivamente) sem diferenÃas entre os gsp+ e gsp-. Houve hiperexpressÃo do miR-21 (p=0,02;fold=10,18) em somatotrofinomas (20,16Â18,48) quando comparado com as HN (2,52 Â3,56), sem diferenÃa entre os gsp + e gsp-. NÃo houve diferenÃa na expressÃo entre os grupos gsp+ e gsp- para os miRNAs let-7a, miR-21, miR-143, miR-15a, miR-23a e miR-24-2. Entretanto, miR-145 e miR-23b foram mais hipoexpressos no grupo gsp+ quando comparados ao gsp- (p=0,03, fold=-4,83 e p=0,02, fold=-2,77, respectivamente). NÃo houve associaÃÃo entre a expressÃo do BTG2 e o painel de miRNAs com agressividade e com o controle da doenÃa. ConclusÃo: Na presente sÃrie de somatotrofinomas, assumidos como esporÃdicos, a frequÃncia de mutaÃÃes nos genes GNAS (35%) e AIP (3,8%) foram semelhantes aos relatados na literatura. NÃo houve diferenÃas nas caracterÃsticas clÃnicas e bioquÃmicas, agressividade, resposta Ãs terapÃuticas, e na expressÃo diferencial do GNAS entre os pacientes com tumores gsp+ e gsp-. HipoexpressÃo de BTG2 (gene supressor tumoral relacionado Ãs vias de sinalizaÃÃo do p53 e do Rb), baixa expressÃo de miRNAs (supressores tumorais) e alta expressÃo de oncomirs em somatotrofinomas sugerem um papel desses na tumorigÃnese somatotrÃfica. / Introduction: Mutations in GNAS and AIP genes are present in 35% and 3%, respectively, of the sporadic somatotropinomas. Recently, increased biological importance of microRNAs (miRNAs) has been observed in pituitary tumorigenesis. However, the molecular mechanisms involved in the pathogenesis of 60% of these tumors remain to be elucidated. Objectives: To identify the prevalence of mutations in GNAS and AIP genes in a series of sporadic somatotropinomas. Compare clinical, bioquimical parametrer at diagnosis as age, tumor size and theirs aggressiveness, pre-operative growth hormone (GH), prolactin (PRL) and insulin-like growth factor-I (IGF-1) levels and treatment responsiveness between somatotropinomas with (gsp+) and without (gsp-) GNAS mutation.To analyze the expression of GNAS and BTG2 genes and a panel of miRNAs between somatotrofinomas and normal pituitaries (NP) and the association between the expression of these genes and miRNAs with aggressiveness, as well as disease control with surgery or control with all adjuvant therapeutic approaches. Material and Methods: 26 patients with acromegaly. GH basal &#8804;2.5&#956;g/L or nadir after OGTT &#8804;1&#956;g/L and normal IGF-I matched for age and sex were used as diagnosis and for cure criteria after transsphenoidal surgery (TS). As control after somatostatin analogues (SA), we adopted the normalization of IGF-I matched for age and sex. Tumor size was evaluated by MRI/CT and the degree of invasiveness by Hardy score (I to IV).Tumor samples (26) were obtained during TS, processed for histopathology and stored at -70ÂC for molecular studies. NP (07) were obtained during autopsy. Total DNA and RNA were extracted by TRIzolÂ. Codons 201 and 227 of the GNAS gene and the whole AIP gene were sequenced. Relative expression of BTG2 and GNAS genes and miRNAs let-7a, miR-16a, miR-21, miR-141, miR-143, miR-15a, miR-145, miR-23a, miR-23b, and miR-24-2 was measured by qPCR (TaqMan probes) using 2-&#916;&#916;Ct method. Results: Frequencies of GNAS and AIP mutations were 35% and 3.8%, respectively. There was no difference between the mean age (39.0  11.5 vs 43.6  9.0 years, p=0.32), basal GH (62.4Â128.1 vs 39.9  48.3 &#956;g/L; p=0.39), IGF-I (435.5  230.8 vs. 556.9  238.3; p=0.32) and PRL (25.7  29.8 vs. 30.9  32.8 ng/L, p=0.69) in plasma concentration, and tumor aggressiveness (p=1.00) between (gsp+) and (gsp-) groups. We observed that 80% (04/05) of gsp+ whereas 33% (02/06) of the gsp- achieved control (p=0.07) after SA therapy adjuvant to TS. When SA, dopamine agonists and/or external radiotherapy were associated 100% (05/05) of gsp+ group and 44% (04/09) of gsp- group (p=0.08) showed disease control.There was no difference in GNAS expression between somatotropinomas and NP (1.07  0.55 vs 0.98  0.28, p=0.97) as well as between somatotropinomasgsp+ and gsp- (1.04  0.59 vs 1.10  0.55, p=0.97, respectively). Hardy I/II tumors showed higher GNAS expression than Hardy III/IV (p=0.02), but there was no association between GNAS expression and disease control with surgery alone or associated with other adjuvant therapies. We observed hypoexpression of BTG2 and miR-16a and miR-141 in somatotropinomas compared with NP (-6.6 fold, p=0.002; -10.0 fold, p=0.01; and -50.0 fold, p=0.0003, respectively) with no difference between gsp+ and gsp- somatotropinomas. There was miR-21 overexpression in somatotropinomas compared with NP (20.2  18.5 vs 2.5  3.6; 10.2 fold, p=0.02), with no difference between gsp+ and gsp- somatotropinomas. However, miR-145 and miR-23b were more hipoexpressed in gsp+ compared to gsp- (-4.8fold, p=0.03 and-2.7 fold, p=0.02). There was no association between the expression of BTG2 and a panel of miRNAs with aggressiveness or disease control. Conclusion: In this series of assumed sporadic somatotopinomas, the frequencies of mutations in GNAS (35%) and AIP (3.8%) were similar to the literature. There were no differences in clinical and biochemical characteristics, aggressiveness, response to therapy, and GNAS expression in patients with gsp+ and gsp- somatotropinomas. Hypoexpression of BTG2, a tumor suppressor gene related to p53 and Rb signaling pathways, low expression of tumor suppressor miRNAs and high expression of oncomirs in somatotropinomas suggest a role in the somatotrophic tumorigenesis.

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior / The voice on congenital isolated growth hormone deficiency (IGHD) is commonly refered high pitched voice . However, vocal and laryngeal data are rare within literature. In Itabaianinha County, Northeast Brazil, it had been described the most extend kindred with severe IGHD due to a GH-realising hormone receptor (GHRHR) gene mutation. The aim of this descriptive transversal study was to evaluate vocal and laryngeal affections in this group. 23 subjects with IGHD were studied, being 6 males and 17 females They were submitted to an otorhinolaryngological evaluation and strobovideolaryngoscopic exam. Besides, a speech evaluation like hearing perception analysis (GRBAS scale) and objective voice evaluation (maximum phonatory time, s/z ratio and acoustic analysis) were also used. The results were performed using qui-square, exact Fisher and unpaired t test (p<0,05). The main vocal complaints were vocal fatigue (36,36%), hoarsenesses (31,81%), sore throat (27,27%), neckache (27,27%), phonatory pain (22,72%) and aphonia (18,18%). No male presented any complaint. Based on findings, vocal abuse (72,72%), etilism (45,45%) and smoking (36,36%) were considered like nocive habits to normal voice. 72,7% of the subjects presented abnormal voice and 27,3% were the normal ones. Amongst the abnormal voice, the mild/moderate grade was prevalent, with the presence of roughness, breathiness and strain. The strobovideoalaryngoscopic exam in 21 subjects showed laryngopharyngeal reflux signs (66,66%), functional dysphonia (52,38%), vocal nodules (23,80%) and vocal cysts (9,52%). There was significant association between vocal complaints and grade of abnormal voice. There weren t association between abnormal voice and laryngeal diseases; nocive habits and abnormal voice; nocive habits and laryngeal diseases; laryngopharyngeal reflux signs and functional dysphonia; vocal complaint and laryngeal diseases. The maximum phonatory time was lower than 10s and s/z ratio was 1,19. In acoustic analysis, fundamental frequency was 221,06 Hz in females and 204,67 Hz in males. There wasn t statistical dependency between genders showing high pitch and dominance IGHD effect above gender. There wasn t statistical dependency between normal and abnormal voice in acoustic analysis neither. Conclusions: Voice presents high pitch in both genders and the most commonly abnormalities were roughness, breathiness and strain. The mild/moderate grade was prevalent. Laryngopharyngeal reflux signs and functional dysphonia were the most reached diagnostics using strobovideolaryngoscopic exam. / A voz na deficiência isolada do hormônio de crescimento (DIGH) de início na infância é comumente referida como voz de caráter agudo, porém dados laríngeos e vocais são escassos na literatura. Na cidade de Itabaianinha-SE, no nordeste brasileiro, foi descrito o maior agrupamento familiar com DIGH severa, devido a uma mutação no gene do receptor do hormônio liberador do hormônio do crescimento (GHRHR). O objetivo do presente estudo foi avaliar alterações laríngeas e vocais nessa população, realizando-se um estudo transversal descritivo. Foram, então, estudados 23 indivíduos com DIGH, sendo 6 do sexo masculino e 17 do sexo feminino. Eles foram submetidos à avaliação otorrinolaringológica geral por entrevista e exame videolaringoestroboscópico. Avaliou-se também o grau de alteração na qualidade vocal, através da análise perceptivo-auditiva (escala GRBAS) e avaliação objetiva da voz (tempo máximo de fonação, relação s/z e análise acústica). Para comparação entre os achados, utilizou-se teste do quiquadrado e exato de Fisher (p<0,05) e entre os sexos, na análise acústica, teste t para amostras independentes (p<0,05). As principais queixas vocais relatadas foram cansaço vocal (36,36%), rouquidão (31,81%), pigarro (27,27%), dor cervical (27,27%), dor à fonação (22,72%) e perda da voz (18,18%). Nenhum indivíduo do sexo masculino apresentou queixa. Os principais hábitos nocivos à saúde vocal foram abuso vocal (72,72%), etilismo (45,45%), tabagismo (36,36%). Foram detectadas 72,7% de vozes alteradas e 27,3% de vozes normais. Dentre as alteradas, o grau leve/moderado foi preponderante, com presença de rouquidão/aspereza, soprosidade e tensão. A videolaringoestroboscopia foi realizada em 21 sujeitos e evidenciou sinais sugestivos de refluxo laringo-faríngeo (66,66%), disfonia funcional (52,38%), nódulos (23,80%) e cisto (9,52%). Houve também associação significativa entre queixas vocais e grau de alteração vocal. Porém, não houve associação significativa entre tipo de voz e doenças detectadas; hábitos nocivos e tipo de voz; hábitos nocivos e doenças detectadas; sinais sugestivos de refluxo laringofaríngeo e disfonia funcional; queixa vocal e doença. Na avaliação objetiva da voz, o tempo máximo de fonação foi abaixo de 10s em todas as variáveis e relação s/z de 1,19 . Na análise acústica, a freqüência fundamental foi 221,06 Hz, em mulheres, e 204,67 Hz, nos homens, sem diferença estatística, caracterizando um patamar agudo e um efeito dominante da DIGH sobre o sexo, nesta variável. Também não houve diferença estatística entre tipo de voz alterada e normal, nas variáveis da análise acústica. Conclusões: a voz foi aguda em ambos os sexos, sendo as alterações vocais mais freqüentes rouquidão/aspereza, soprosidade e tensão, grau leve/moderado. Os sinais sugestivos de refluxo laringo-faríngeo e disfonia funcional foram os achados mais comuns na videolaringoestroboscopia.

Voice is produced by the vibration of the vocal folds expressed by its fundamental frequency (f0 Hz), while the formants (F) are f0 multiples, indicating amplification zones of the vowels in the vocal tract. The isolated growth hormone (GH) deficiency (IGHD) characterizes high pitch voice, with higher values of formants frequencies, maintaining a prepuberal acoustic prediction. We speculated that acoustic vocal parameters can be improved by SOVTT or choir training, and that their voice suits distribution resembles an infantile choir. Objectives: verify the effect of therapy with semi-fluid vocal tract exercise (SOVTT); evaluate the effect of coral training; and to analyze the classification of voice suits in the choir of subjects with DIGH. METHODS: prospective longitudinal study without a control group evaluated the effects of SOVTT therapy on LaxVox silicone tubing and coral training on 17 adults with congenital, untreated IGHD individuals over a 30-day period. The vocal sample of the vowel [é] was recorded for acoustic analysis and comparison at three moments of the effect before and after the SOVTT (pre- SOVTT and post- SOVTT) and post-training choir. In addition, classification of the voices of the choir voices of individuals with IGHD was performed. Results: The first formant (F1) was higher in moment post-training compared to the pre- SOVTT (p = 0.009). The second formant (F2) was higher in post-SOVTT in comparison to pre- SOVTT (p= 0.045). Shimmer was reduced in post-training in comparison to pre-SOVTT (p= 0.045). All ten IGHD women were all altos, and all seven IGHD men were tenors. Conclusions: An approach to speech therapy with SOVTT and choir training was able to improve acoustic parameters of the voice of individuals with congenital and untreated IGHD. In particular, this appears to be important in a scenario in which few patients undergo GH replacement therapy. It is concluded that the effect of the therapy with SOVTT is added to coral training favoring adjustments of the source in the adduction of the vocal folds and the filter in the form, length and constriction of the vocal tract. The classification of voices of the choir voices of adult individuals IGHD is typical of an infantile choir. / A voz é produzida pela vibração das pregas vocais, cujo número de ciclos por segundo (hertz, Hz) corresponde à frequência fundamental (f0) do sinal laríngeo, e os formantes (F) indicam as zonas de amplificação das vogais no trato vocal. A deficiência do hormônio de crescimento (GH) isolada (DIGH) caracteriza a voz de timbre alto e pitch agudo, com valores elevados de frequências de formantes, mantendo uma previsão acústica pré-puberal. Especula-se que as medidas acústicas vocais desses indivíduos podem ser melhoradas pelo exercício de trato vocal semiocluído e treinamento de coral. Objetivos: avaliar o efeito da terapia com exercício de trato vocal semiocluído (ETVSO); avaliar o efeito do treinamento de coral nesses indivíduos com DIGH. Métodos: trata-se de um estudo prospectivo longitudinal sem grupo controle. A análise acústica da vocal da vogal [é] foi realizada em 17 indivíduos adultos de DIGH antes e após do ETVSO (pré-ETVSO e pós-ETVSO) e pós-treinamento de coral, em um período de 30 dias. O coral DIGH adulto de ambos os sexos se assemelha a um coral infantil com classificação dos naipes de vozes graves e agudas. Resultados: O primeiro formante (F1) foi maior pós-treinamento de coral em comparação ao pré-ETVSO (p = 0,009). O segundo formante (F2) foi maior no pós-ETVSO em comparação ao pré-ETVSO (p = 0,045). Houve uma tendência a redução do shimmer pós-treinamento de coral em comparação ao pré-ETVSO (p = 0,051) e uma redução no treinamento pós-coro em comparação com pós- ETVSO (p = 0,047). Conclusão: a terapia com ETVSO foi relevante para F2 e treinamento de coral melhorou a F1 e reduziu o shimmer. Portanto, esta abordagem de terapia fonoaudiológica foi capaz de melhorar os parâmetros acústicos da voz de indivíduos com DIGH congênita e não tratada. Isso parece particularmente importante em um cenário em que poucos pacientes são submetidos à terapia de reposição de GH. / Aracaju, SE

Many genes demonstrate highly dynamic pulsatile expression, with characteristic bursts of activity. Dynamic expression of the human prolactin (hPrl) gene in pituitary cells has previously been investigated identifying key temporal characteristics, influenced by the process of chromatin remodelling. Earlier work on the related pituitary human growth hormone (hGH) proximal promoter (-496/+1bp) indicated that it displayed similar dynamic behaviour. The human GH gene contains an extensive long-distance regulatory sequence, including a locus control region (-14/-32kbp) that has been shown to regulate chromatin remodelling and confer tissue-specificity of hGH expression. In this work I aimed to study dynamic regulation of the hGH gene promoter in detail. Initially I investigated the efficiency of several methods to express the luciferase gene in a 180kb hGH genomic fragment using bacterial artificial chromosome recombineering, to allow the investigation of single cell transcription dynamics. Although a functional recombinant BAC was not finalised during the course of the work, I carried out detailed time course studies using shorter hGH-reporter constructs. Using quantitative microscopy to study live single cells, I compared the dynamic characteristics of a 5kb hPrl promoter fragment with those of -840/+1bp and -3348/+1bp hGH-luciferase promoter-reporter constructs. Whilst previous hPrl analysis utilised a binary mathematical model assuming a simplified two-state (ON/OFF) process of gene transcription, I validated and applied a novel stochastic switch model (SSM), assuming instead that transcription rate can switch between any variable states at any time. Through doing so I observed an asymmetry in transcription rate switching, suggesting an all-or-nothing activation of a single UP-switch, with a greater number of rate decreasing DOWN-switches. The -3348/+1bp construct produced double the number of DOWN-switches, whilst the -840/+1bp construct produced 1.5 DOWN-switches in a 48h period. The cycling of transcriptional activity seen by the shorter construct was modified through the addition of forskolin, activating cAMP signalling. However, significant modification of the transcriptionally inactive refractory period seen with the -3348/+1bp construct (reduced from 3h to 1.9h) required histone modification through application of trichostatin A, a HDAC inhibitor. In conclusion, different promoter elements confer different transcriptional timing and dynamics. A subtler transcriptional modelling, such as used here in the SSM, reveals new insights into the phenomena of transcriptional switching, but the mechanisms involved remain to be determined.

We used the GENECONV program, the Hsu et al. (2010) method and phylogenetic analyses to analyze the gene conversions which occurred in the growth hormone, folate receptor and trypsin gene families of six primate species. Significant positive correlations were found between sequence similarity and conversion length in all but the trypsin gene family. Converted regions, when compared to non-converted ones, also displayed a significantly higher GC-content in the growth hormone and folate receptor gene families. Finally, all detected gene conversions were found to be less frequent in conserved gene regions and towards functionally important genes. This suggests that purifying selection is eliminating all gene conversions having a negative functional impact.