A measurement of the proton elastic form factors for 1 ��� Q^2 ��� 3 (GeV/c)^2

We report measurements of the proton form factors, G^p_E and G^p_M, extracted from elastic electron scattering in the range 1 ��� Q^2 ��� 3 (GeV/c)^2 with uncertainties of <15% in G^p_E and <3% in G^p_M. The results for G^p_E are somewhat larger than indicated by most theoretical parameterizations. The ratio of Pauli and Dirac form factors, Q^2(F^p_2/F^p_1), is lower in value and demonstrates less Q^2 dependence than these parameterizations have indicated. Comparisons are made to theoretical models, including those based on perturbative QCD, vector-meson dominance, QCD sum rules, and diquark constituents to the proton. A global extraction of the form factors, including previous elastic scattering measurements, is also presented.

Sequence specific complexation of b DNA at sites containing g,c base pairs

<p>A series of eight related analogs of distamycin A has been synthesized. Footprinting and affinity cleaving reveal that only two of the analogs, pyridine-2- car box amide-netropsin (2-Py N) and 1-methylimidazole-2-carboxamide-netrops in (2-ImN), bind to DNA with a specificity different from that of the parent compound. A new class of sites, represented by a TGACT sequence, is a strong site for 2-PyN binding, and the major recognition site for 2-ImN on DNA. Both compounds recognize the G���C bp specifically, although A's and T's in the site may be interchanged without penalty. Additional A���T bp outside the binding site increase the binding affinity. The compounds bind in the minor groove of the DNA sequence, but protect both grooves from dimethylsulfate. The binding evidence suggests that 2-PyN or 2-ImN binding induces a DNA conformational change.</p> <p>In order to understand this sequence specific complexation better, the Ackers quantitative footprinting method for measuring individual site affinity constants has been extended to small molecules. MPE���Fe(II) cleavage reactions over a 10^5 range of free ligand concentrations are analyzed by gel electrophoresis. The decrease in cleavage is calculated by densitometry of a gel autoradiogram. The apparent fraction of DNA bound is then calculated from the amount of cleavage protection. The data is fitted to a theoretical curve using non-linear least squares techniques. Affinity constants at four individual sites are determined simultaneously. The distamycin A analog binds solely at A���T rich sites. Affinities range from 10^(6)- 10^(7)M^(-1) The data for parent compound D fit closely to a monomeric binding curve. 2-PyN binds both A���T sites and the TGTCA site with an apparent affinity constant of 10^(5) M^(-1). 2-ImN binds A���T sites with affinities less than 5 x 10^(4) M^(-1). The affinity of 2-ImN for the TGTCA site does not change significantly from the 2-PyN value. At the TGTCA site, the experimental data fit a dimeric binding curve better than a monomeric curve. Both 2-PyN and 2-ImN have substantially lower DNA affinities than closely related compounds.</p> <p>In order to probe the requirements of this new binding site, fourteen other derivatives have been synthesized and tested. All compounds that recognize the TGTCA site have a heterocyclic aromatic nitrogen ortho to the N or C-terminal amide of the netropsin subunit. Specificity is strongly affected by the overall length of the small molecule. Only compounds that consist of at least three aromatic rings linked by amides exhibit TGTCA site binding. Specificity is only weakly altered by substitution on the pyridine ring, which correlates best with steric factors. A model is proposed for TGTCA site binding that has as its key feature hydrogen bonding to both G's by the small molecule. The specificity is determined by the sequence dependence of the distance between G's.</p> <p>One derivative of 2-PyN exhibits pH dependent sequence specificity. At low pH, 4-dimethylaminopyridine-2-carboxamide-netropsin binds tightly to A���T sites. At high pH, 4-Me_(2)NPyN binds most tightly to the TGTCA site. In aqueous solution, this compound protonates at the pyridine nitrogen at pH 6. Thus presence of the protonated form correlates with A���T specificity.</p> <p>The binding site of a class of eukaryotic transcriptional activators typified by yeast protein GCN4 and the mammalian oncogene Jun contains a strong 2-ImN binding site. Specificity requirements for the protein and small molecule are similar. GCN4 and 2-lmN bind simultaneously to the same binding site. GCN4 alters the cleavage pattern of 2-ImN-EDTA derivative at only one of its binding sites. The details of the interaction suggest that GCN4 alters the conformation of an AAAAAAA sequence adjacent to its binding site. The presence of a yeast counterpart to Jun partially blocks 2-lmN binding. The differences do not appear to be caused by direct interactions between 2-lmN and the proteins, but by induced conformational changes in the DNA protein complex. It is likely that the observed differences in complexation are involved in the varying sequence specificity of these proteins.</p>

Structurally engineered cytochromes c with novel ligand-binding properties

<p>Semisynthesis of horse heart cytochrome c and site-directed mutagenesis of Saccharomyces cerevisiae (S. c.) iso-1-cytochrome c have been utilized to substitute Ala for the cytochrome c heme axial ligand Met80 to yield ligand-binding proteins (horse heart Ala80cyt c and S.c. Ala80cyt c) with spectroscopic properties remarkably similar to those of myoglobin. Both species of Fe(II)Ala80cyt c form exceptionally stable dioxygen complexes with autoxidation rates 10-30x smaller and O₂ binding constants ~ 3x greater than those of myoglobin. The resistance of O₂-Fe(II)Ala80cyt c to autoxidation is attributed in part to protection of the heme site from solvent as exhibited by the exceptionally slow rate of CO binding to the heme as well as the low quantum yield of CO photodissociation.</p> <p>UV/vis, EPR, and paramagnetic NMR spectroscopy indicate that at pH 7 the Fe(III)Ala80cyt c heme is low-spin with axial His-OH^- coordination and that below pH ~6.5, Fe(III)Ala80cyt cis high-spin with His-H₂O heme ligation. Significant differences in the pH dependence of the ¹H NMR spectra of S.c. Fe(III)Ala80cyt c compared to wild-type demonstrate that the axial ligands influence the conformational energetics of cytochrome c.</p> <p>¹H NMR spectroscopy has been utilized to determine the solution structure of the cyanide derivative of S.c. Fe(III)Ala80cyt c. 82% of the resonances in the ¹H NMR spectrum of S.c. CN-Fe(III)Ala80cyt c have been assigned through 1D and 2D experiments. The RMSD values after restrained energy minimization of the family of 17 structures obtained from distance geometry calculations are 0.68 �� 0.11 �� for the backbone and 1.32 �� 0.14 �� for all heavy atoms. The solution structure indicates that a tyrosine in the "distal" pocket of CN-Fe(III)Ala80cyt c forms a hydrogen bond with the Fe(III)-CN unit, suggesting that it may play a role analogous to that of the distal histidine in myoglobin in stabilizing the dioxygen adduct.</p>

Charge symmetry in ^(13)N and ^(13)C a coupled-channel model

A set of coupled-channel differential equations based on a rotationally distorted optical potential is used to calculate the wave functions required to evaluate the gamma ray transition rate from the first excited state to the ground state in ^(13)C and ^(13)N. The bremsstrahlung differential cross section of low energy protons is also calculated and compared with existing data. The marked similarity between the potentials determined at each resonance level in both nuclei supports the hypothesis of the charge symmetry of nuclear forces by explaining the deviation of the ratios of the experimental E1 transition strengths from unity.

Comparative studies of Vulva development in C. briggsae

<p>As evolution progresses, developmental changes occur. Genes lose and gain molecular partners, regulatory sequences, and new functions. As a consequence, tissues evolve alternative methods to develop similar structures, more or less robust. How this occurs is a major question in biology. One method of addressing this question is by examining the developmental and genetic differences between similar species. Several studies of nematodes Pristionchus pacificus and Oscheius CEW1 have revealed various differences in vulval development from the well-studied C. elegans (e.g. gonad induction, competence group specification, and gene function.)</p> <p>I approached the question of developmental change in a similar manner by using Caenorhabditis briggsae, a close relative of C. elegans. C. briggsae allows the use of transgenic approaches to determine developmental changes between species. We determined subtle changes in the competence group, in 1�� cell specification, and vulval lineage.</p> <p>We also analyzed the let-60 gene in four nematode species. We found conservation in the codon identity and exon-intron boundaries, but lack of an extended 3' untranslated region in Caenorhabditis briggsae.</p>

Signal transduction, regulation, and developmental logic of EGFR signalling in C. elegans

<p>RTKs-mediated signaling systems and the pathways with which they interact (e.g., those initiated by G protein-mediated signaling) involve a highly cooperative network that sense a large number of cellular inputs and then integrate, amplify, and process this information to orchestrate an appropriate set of cellular responses. The responses include virtually all aspects of cell function, from the most fundamental (proliferation, differentiation) to the most specialized (movement, metabolism, chemosensation). The basic tenets of RTK signaling system seem rather well established. Yet, new pathways and even new molecular players continue to be discovered. Although we believe that many of the essential modules of RTK signaling system are rather well understood, we have relatively little knowledge of the extent of interaction among these modules and their overall quantitative importance.</p> <p>My research has encompassed the study of both positive and negative signaling by RTKs in C. elegans. I identified the C. elegans S0S-1 gene and showed that it is necessary for multiple RAS-mediated developmental signals. In addition, I demonstrated that there is a SOS-1-independent signaling during RAS-mediated vulval differentiation. By assessing signal outputs from various triple mutants, I have concluded that this SOS-1-independent signaling is not mediated by PTP-2/SHP-2 or the removal of inhibition by GAP-1/ RasGAP and it is not under regulation by SLI-1/Cb1. I speculate that there is either another exchange factor for RASor an as yet unidentified signaling pathway operating during RAS-mediated vulval induction in C. elegans.</p> <p>In an attempt to uncover the molecular mechanisms of negative regulation of EGFR signaling by SLI-1/Cb1, I and two other colleagues codiscovered that RING finger domain of SLI-1 is partially dispensable for activity. This structure-function analysis shows that there is an ubiquitin protein ligase-independent activity for SLI-1 in regulating EGFR signaling. Further, we identified an inhibitory tyrosine of LET-23/ EGFR requiring sli-1(+)for its effects: removal of this tyrosine closely mimics loss of sli-1 but not loss of other negative regulator function.</p> <p>By comparative analysis of two RTK pathways with similar signaling mechanisms, I have found that clr-1, a previously identified negative regulator of egl-15 mediated FGFR signaling, is also involved in let-23 EGFR signaling. The success of this approach promises a similar reciprocal test and could potentially extend to the study of other signaling pathways with similar signaling logic.</p> <p>Finally, by correlating the developmental expression of lin-3 EGF to let-23 EGFR signaling activity, I demonstrated the existence of reciprocal EGF signaling in coordinating the morphogenesis of epithelia. This developmental logic of EGF signaling could provide a basis to understand a universal mechanism for organogenesis.</p>

Utilidad de la biblioteca escolar como recurso al servicio del proyecto educativo

RESUMEN: En este trabajo se plasma la transcendencia que tiene la lectura en el desarrollo integral de las personas. Del mismo modo, consideramos fundamental que se fomente la lectura en la escuela desde la etapa de infantil. El supuesto de nuestro trabajo es que la escuela cuenta con un espacio que contiene todo lo necesario para fomentar el h��bito lector: la biblioteca escolar. Para nuestra investigaci��n nos hemos acercado a un centro escolar de Amorebieta (Bizkaia) para conocer si el funcionamiento de las bibliotecas escolares se realiza siguiendo los patrones que se consideran adecuados desde la literatura especializada en el tema.

Ancillary ligand effects : from zirconium(IV)-catalyzed homogeneous propylene polymerization to platinum(II)-mediated C-H bond activation

<p>A series of C_s- and C₁-symmetric doubly-linked ansa-metallocenes of the general formula {1,1'-SiMe₂-2,2'-E-('��⁵-C₅H₂-4-R¹)-(��⁵-C₅H-3',5'-(CHMe₂)₂)}ZrC₂ (E = SiMe₂ (1), SiPh₂ (2), SiMe₂ -SiMe₂ (3); R¹ = H, CHMe₂, C₅H₉, C₆H₁₁, C₆H₅) has been prepared. When activated by methylaluminoxane, these are active propylene polymerization catalysts. 1 and 2 produce syndiotactic polypropylenes, and 3 produces isotactic polypropylenes. Site epimerization is the major pathway for stereoerror formation for 1 and 2. In addition, the polymer chain has slightly stronger steric interaction with the diphenylsilylene linker than with the dimethylsilylene linker. This results in more frequent site epimerization and reduced syndiospecificity for 2 compared to 1. </p> <p>C₁-Symmetric ansa-zirconocenes [1,1 '-SiMe₂-(C₅H₄)-(3-R-C₅H₃)]ZrCl₂ (4), [1,1 '-SiMe₂-(C₅H₄)-(2,4-R₂-C₅H₂)]ZrCl₂ (5) and [1,1 '-SiMe₂-2,2 '-(SiMe₂-SiMe₂)-(C₅H₃)-( 4-R-C₅H₂)]ZrCl₂ (6) have been prepared to probe the origin of isospecificity in 3. While 4 and 3 produce polymers with similar isospecificity, 5 and 6 give mostly hemi-isotactic-like polymers. It is proposed that the facile site epimerization via an associative pathway allows rapid equilibration of the polymer chain between the isospecific and aspecific insertion sites. This results in more frequent insertion from the isospecific site, which has a lower kinetic barrier for chain propagation. On the other hand, site epimerization for 5 and 6 is slow. This leads to mostly alternating insertion from the isospecific and aspecific sites, and consequently, a hemi-isotactic-like polymers. In comparison, site epimerization is even slower for 3, but enchainment from the aspecific site has an extremely high kinetic barrier for monomer coordination. Therefore, enchainment occurs preferentially from the isospecific site to produce isotactic polymers. </p> <p>A series of cationic complexes [(ArN=CR-CR=NAr)PtMe(L)]⁺[BF₄]⁺ (Ar = aryl; R = H, CH₃; L = water, trifluoroethanol) has been prepared. They react smoothly with benzene at approximately room temperature in trifluoroethanol solvent to yield methane and the corresponding phenyl Pt(II) cations, via Pt(IV)-methyl-phenyl-hydride intermediates. The reaction products of methyl-substituted benzenes suggest an inherent reactivity preference for aromatic over benzylic C-H bond activation, which can however be overridden by steric effects. For the reaction of benzene with cationic Pt(II) complexes, in which the diimine ligands bear 3,5-disubstituted aryl groups at the nitrogen atoms, the rate-determining step is C-H bond activation. For the more sterically crowded analogs with 2,6-dimethyl-substituted aryl groups, benzene coordination becomes rate-determining. The more electron-rich the ligand, as reflected by the CO stretching frequency in the IR spectrum of the corresponding cationic carbonyl complex, the faster the rate of C-H bond activation. This finding, however, does not reflect the actual C-H bond activation process, but rather reflects only the relative ease of solvent molecules displacing water molecules to initiate the reaction. That is, the change in rates is mostly due to a ground state effect. Several lines of evidence suggest that associative substitution pathways operate to get the hydrocarbon substrate into, and out of, the coordination sphere; i.e., that benzene substitution proceeds by a solvent- (TFE-) assisted associative pathway. </p>

Modelagem do potencial el��trico atrav��s da membrana do neur��nio ganglionar e c��lulas de neuroblastoma: efeitos das cargas superficiais.

O objetivo do presente trabalho �� comparar, do ponto de vista el��trico, a membrana do neur��nio ganglionar com a da c��lula de neuroblastoma, analisando os efeitos das cargas fixas sobre o potencial el��trico nas superf��cies da bicamada lip��dica e tamb��m sobre o comportamento do perfil de potencial atrav��s da membrana, considerando as condi����esf��sico-qu��micas do estado de repouso e do estado de potencial de a����o. As condi����es para a ocorr��ncia dos referidos estados foram baseadas em valores num��ricos de par��metros el��tricos e qu��micos, caracter��sticos dessas c��lulas, obtidos na literatura. O neur��nio ganglionar exemplifica um neur��nio sadio, e a c��lula de neuroblastoma, que �� uma c��lula tumoral, exemplifica um neur��nio patol��gico, alterado por esta condi����o. O neuroblastoma �� um tumor que se origina das c��lulas da crista neural (neuroblastos), que �� uma estrutura embrion��ria que d�� origem a muitas partes do sistema nervoso, podendo surgir em diversos locais do organismo, desde a regi��o do cr��nio at�� a ��rea mais inferior da coluna. O modelo adotado para simular a membrana de neur��nio inclui: (a) as distribui����es espaciais de cargas el��tricas fixas no glicoc��lix e na rede de prote��nas citoplasm��ticas; (b) as distribui����es de cargas na solu����o eletrol��tica dos meios externo e interno; e (c) as cargas superficiais da bicamada lip��dica. Os resultados que obtivemos mostraram que, nos estados de repouso e de a����o, os potenciais superficiais da bicamada interno (��Sbc) e externo (��Sgb) da c��lula de neuroblastoma n��o sofrem altera����o mensur��vel, quando a densidade de carga na superf��cie interna (QSbc) torna-se 50 vezes mais negativa, tanto para uma densidade de carga na superf��cie externa da bicamada nula (QSgb = 0), como para um valor de QSgb 6= 0. Por��m, no estado de repouso, uma leve queda em ��Sbc do neur^onio ganglionar pode ser observada com este n��vel de varia����o de carga, sendo que ��Sgb do neur��nio ganglionar �� mais negativo quando QSgb = 1=1100 e/A2. No estado de a����o, para QSgb = 0, o aumento da negatividade de QSbc n��o provoca altera����o detect��vel de ��Sbc e ��Sgb para os dois neur��nios. Quando consideramos QSgb = 1=1100 e/A2, ��Sgb do neur��nio ganglionar se torna mais negativo, n��o se observando varia����es detect��veis nos potenciais superficiais da c��lula de neuroblastoma. Tanto no repouso quanto no estado de a����o, ��Sgb das duas c��lulas n��o sofre varia����o sens��vel com o aumento da negatividade da carga fixa distribu��da espacialmente no citoplasma. J�� a ��Sbc sofre uma queda gradativa nos dois tipos celulares; por��m, no estado de a����o, esta queda �� mais r��pida. Descobrimos diferen��as importantes nos perfis de potencial das duas c��lulas, especialmente na regi��o do glicoc��lix.

Regula����o end��crina de curto prazo de horm��nios relacionados �� fome em mulheres obesas que apresentam epis��dios de compuls��o alimentar

A compuls��o alimentar est�� associada a diversas doen��as, entre elas, a obesidade.Com o intuito de pesquisar a diferen��a hormonal ligada ao controle da fome e da saciedade associada ao epis��dio de compuls��o alimentar (ECA), avaliou-se a concentra����o s��rica dos horm��nios que regulam este processo em mulheres adultas. M��todos: O estudo experimental foi composto de 3 grupos (n=23), sendo: grupo Eutr��fico (GE;n=8), grupo obeso sem ECA (GO;n=7) e obesas com ECA (ECA;n=8). Todas as mulheres que participaram do estudo freq��entavam os servi��os de sa��de da Policl��nica Piquet Carneiro. Foram dosados os horm��nios: Grelina Total, Glucagon, Adiponectina, Amilina, Pept��deo C, GLP-1, Insulina e Leptina s��ricos nos tempos: jejum, 15 e 60 minutos ap��s a ingest��o da refei����o fornecida. As refei����es ingeridas foram controladas em energia, 55% carboidratos, 15% prote��nas, 30% lip��dios. Os dados foram analisados como valores m��dios por grupo em software SAS, considerando p<0,05. Resultados: A idade das mulheres estudadas variou de 32 a 50 anos. A concentra����o de adiponectina encontrada, que �� inversamente proporcional a adiposidade, foi significativamente menor no grupo ECA em rela����o aos demais grupos (p=0,01). Em rela����o �� leptina, o grupo GO apresentou concentra����o maior em rela����o aos demais grupos (p<0,0001). J��, a concentra����o de grelina encontrada foi significativamente menor no grupo ECA em rela����o aos demais grupos (p=0,02). Foram encontradas concentra����es significativamente maiores de insulina no grupo GO em rela����o aos demais grupos (p=0,04). A concentra����o de amilina encontrada foi significativamente maior no grupo GO em rela����o aos outros grupos (p=0,01). A concentra����o de GLP-1 encontrada no grupo GO foi maior em m��dia, por��m esta diferen��a n��o foi estatisticamente significativa entre os grupos (p=0,25). A concentra����o de Pept��deo C encontrada no grupo GO foi maior em rela����o aos outros grupos (p=0,003). Apesar da concentra����o de Glucagon no grupo ECA ser maior em rela����o aos demais grupos, estes valores n��o eram diferentes estatisticamente (p=0,13). Nossos achados mostraram que obesas ECA tem perfil hormonal diferente de obesas sem ECA. A baixa concentra����o de grelina do grupo de obesas ECA e a alta concentra����o de insulina, pept��deo C e amilina nas obesas com e sem ECA pode estar relacionado com o aumento da ingest��o alimentar e com o desequil��brio energ��tico.

Development of a new modelling framework to estimate the C footprint from Basque dairy farms

19 p.

An��lise das altera����es decorrentes do envelhecimento na mec��nica respirat��ria pela t��cnica de oscila����es for��adas

O crescimento do percentual de idosos na popula����o ocorre mundialmente tornando necess��rio conhecer o impacto do processo de envelhecimento, neste contexto, do sistema respirat��rio. O desconhecimento do impacto do envelhecimento associado a diferentes graus de exposi����o a poluentes e a presen��a de comorbidade(s) dificulta a diagnose das pneumopatias acarretando aos idosos piora da qualidade de vida. S��o vantagens da T��cnica de Oscila����es For��adas (FOT): alto potencial de aplica����o em idosos, f��cil realiza����o, an��lise detalhada da mec��nica respirat��ria, desempenho de papel complementar, bem como de alternativa na impossibilidade de realiza����o dos exames tradicionais. Foi realizado um estudo experimental comparativo que objetivou investigar o impacto do envelhecimento no sistema respirat��rio pela FOT e pela espirometria entre grupos de diferentes faixas et��rias, sendo a idade a vari��vel independente e as vari��veis dependentes, os par��metros oscilom��tricos resist��ncia em regime cont��nuo (R0) e das vias a��reas centrais (Rm), inclina����o da resist��ncia (S), frequ��ncia de resson��ncia (fr), reat��ncia m��dia (Xm), complac��ncia din��mica (Cdin,sr) e os par��metros espirom��tricos (VEF1, CVF, VEF1/CVF e FEF/CVF). Foram realizados entrevista, exame cl��nico, radiografia tor��cica, avalia����o da mec��nica respirat��ria pela FOT e da fun����o pulmonar pela espirometria. 255 indiv��duos com idades entre 20 e 86 anos foram entrevistados. Destes, 175 foram exclu��dos, restando os 80 volunt��rios analisados, que foram divididos em 6 grupos de acordo com a faixa et��ria (A: 20 a 29 anos; B: 30 a 39 anos; C: 40 a 49 anos; D: 50 a 59 anos; E: 60 a 69 anos; F: 70 anos ou mais). Foram utilizados os testes de Shapiro-Wilkins, na avalia����o da normalidade dos dados biom��tricos em cada grupo, Oneway ANOVA, na compara����o entre os grupos, e Tukey HSD na compara����o entre as classes subjacentes. A an��lise da associa����o entre duas vari��veis foi realizada inicialmente pela regress��o univariada entre os par��metros oscilom��tricos, a idade e a altura. A regress��o m��ltipla entre os par��metros oscilom��tricos, idade e altura foi realizada em conjunto. Foi realizada a an��lise de confundimento ou modifica����o de efeito sobre o par��metro altura na rela����o entre a idade e os par��metros oscilom��tricos. A corre����o pelo fator altura foi realizada quando sua an��lise apresentava fator de confundimento. Quanto aos par��metros resistivos, n��o foram observadas altera����es em R0 e Rm com o envelhecimento enquanto que o decl��nio observado em S �� discreto e n��o-significativo. Em rela����o aos par��metros reativos, verificouse que Cdin,sr e Xm diminuem enquanto que fr aumenta com o processo de envelhecimento. Todas estas altera����es s��o significativas. Todavia, a diminui����o da Cdin,sr n��o apresenta rela����o com a idade e sim com a altura, que constituiu modifica����o do efeito. Nos demais par��metros oscilom��tricos, a altura constituiu fator de confundimento. Quanto �� espirometria, observou-se decl��nio significativo do VEF1, do VEF1/CVF e da CVF. O ��ndice FEF/CVF apresentou decl��nio n��osignificativo. Concluindo, a resist��ncia do sistema respirat��rio e a complac��ncia din��mica n��o se modificam enquanto a homogeneidade do sistema respirat��rio diminui com o processo de envelhecimento.

Avalia����o da fun����o endotelial microvascular, da rigidez arterial e da resposta inflamat��ria sist��mica em pacientes submetidos a cirurgia card��aca com circula����o extracorp��rea

A Cirurgia de Revasculariza����o do Mioc��rdio, realizada com o aux��lio da Circula����o Extracorp��rea, est�� associada a altera����es importantes na microcircula����o e na produ����o e circula����o de citocinas e marcadores inflamat��rios. No presente estudo, foram avaliados 23 pacientes com indica����o de Revasculariza����o do Mioc��rdio, no dia do procedimento e 7 e 28 dias ap��s a cirurgia. A microcircula����o cut��nea, enquanto reflexo da microcircula����o coronariana, foi estudada atrav��s da hiperemia t��rmica e/ ou reativa p��s oclusiva e da iontoforese de subst��ncias vasoativas por mecanismos dependentes e independentes do endot��lio. A rigidez arterial foi aferida atrav��s da an��lise da onda de pulso digital. Foi avaliado ainda o impacto da doen��a e do procedimento cir��rgico sobre a produ����o e circula����o s��rica de citocinas e marcadores inflamat��rios, tais como: PCR-HS, nitrito/ nitrato, IL-6, Il-7, IL-8, IL-10, IFN-&#947;, TNF-&#945; e G-CSF. Foi observada uma tend��ncia �� redu����o da vasodilata����o da microcircula����o cut��nea ap��s a administra����o de doses acumulativas de acetilcolina (endot��lio dependente) atrav��s da iontoforese de 7 e 28 dias ap��s o procedimento cir��rgico. A hiperemia t��rmica foi mais pronunciada na avalia����o basal do que aos 7 e 28 dias. A hiperemia reativa p��s oclusiva n��o demonstrou altera����es 7 dias ap��s o procedimento. Aos 28 dias, houve um aumento da condut��ncia microvascular cut��nea. Quando avaliada a vasodilata����o endot��lio-independente (nitroprussiato de s��dio), observamos aumento do fluxo microvascular cut��neo diretamente proporcional �� carga/ dose aplicada, sem diferen��as nos valores obtidos no basal e 7 e 28 dias ap��s o procedimento. A rigidez arterial n��o apresentou altera����es. A an��lise dos fatores inflamat��rios e das citocinas demonstrou aumento marcante da IL-6 e da IL-8 ap��s 7 dias do procedimento cir��rgico, com retorno parcial aos n��veis basais da IL-6 e total da IL-8 ap��s 28 dias. O IFN-&#947;, TNF-&#945; e G-CSF apenas apresentaram n��veis detect��veis na avalia����o basal e IL-7 e IL-10 n��o demonstraram altera����es significativas nos tempos avaliados. A PCR-HS demonstrou n��veis mais elevados ap��s 7 dias e retorno parcial aos n��veis basais ap��s 28 dias. O nitrito/ nitrato, ap��s 7 dias, apresentou leve queda em sua concentra����o plasm��tica. Conclu��mos que a pequena diferen��a entre o valores obtidos entre o basal e ap��s 7 dias do procedimento cir��rgico com a iontoforese de acetilcolina resulta em minimiza����o do impacto endotelial e um valor constante deste dado ap��s 28 dias, sugere recomposi����o fisiol��gica completa. Este resultado foi semelhante com a an��lise da hiperemia t��rmica e reativa p��s oclusiva. As interleucinas IL-6 e IL-8, bem como a PCR-HS apresentaram comportamento correlacion��vel, refletindo a cin��tica inflamat��ria. A rigidez arterial n��o demonstrou altera����es.

The C. elegans ALA neuron : its transcriptome and role in inducing sleep

A long-standing yet to be accomplished task in understanding behavior is to dissect the function of each gene involved in the development and function of a neuron. The C. elegans ALA neuron was chosen in this study for its known function in sleep, an ancient but less understood animal behavior. Single-cell transcriptome profiling identified 8,133 protein-coding genes in the ALA neuron, of which 57 are neuropeptide-coding genes. The most enriched genes are also neuropeptides. In combination with gain-of-function and loss-of-function assays, here I showed that the ALA-enriched FMRFamide neuropeptides, FLP-7, FLP-13, and FLP-24, are sufficient and necessary for inducing C. elegans sleep. These neuropeptides act as neuromodulators through GPCRs, NPR-7, and NPR-22. Further investigation in zebrafish indicates that FMRFamide neuropeptides are sleep-promoting molecules in animals. To correlate the behavioral outputs with genomic context, I constructed a gene regulatory network of the relevant genes controlling C. elegans sleep behavior through EGFR signaling in the ALA neuron. First, I identified an ALA cell-specific motif to conduct a genome-wide search for possible ALA-expressed genes. I then filtered out non ALA-expressed genes by comparing the motif-search genes with ALA transcriptomes from single-cell profiling. In corroborating with ChIP-seq data from modENCODE, I sorted out direct interaction of ALA-expressed transcription factors and differentiation genes in the EGFR sleep regulation pathway. This approach provides a network reference for the molecular regulation of C. elegans sleep behavior, and serves as an entry point for the understanding of functional genomics in animal behaviors.