Η Heparin affin regulatory peptide (HARP) είναι ένας αυξητικός παράγοντας με μοριακό βάρος 18 kDa, που έχει μεγάλη συγγένεια με την ηπαρίνη. Είναι συντηρημένη μεταξύ διαφόρων ειδών και παρουσιάζει 50% ομολογία με τη Midkine και την RI-HBP. Οι πρωτεΐνες αυτές συγκροτούν μια σχετικά νέα οικογένεια αυξητικών παραγόντων που έχουν συγγένεια με την ηπαρίνη. Η HARP απομονώθηκε για πρώτη φορά από τον εγκέφαλο νεογέννητου βοός ως ένα μόριο που μπορεί να επάγει την προέκταση των νευρικών κυττάρων. Επίσης, εκφράζεται στη μήτρα, στους χόνδρους και στα οστά. Αρκετές αναφορές αποδεικνύουν ότι υπάρχει μεγάλη συσχέτιση μεταξύ της έκφρασης της HARP και της ανάπτυξης καρκινικού όγκου και της αγγειογένεσης. Η HARP αποτελεί μιτογόνο παράγοντα για διάφορους τύπους ενδοθηλιακών κυττάρων, ενώ μπορεί να επάγει την αγγειογένεση in vivo και in vitro. Ασκεί τη βιολογική της δράση μετά από αλληλεπίδραση με πρωτεογλυκάνες της επιφάνειας του κυττάρου, όπως η N-συνδεκάνη, ή μετά από δέσμευση σε πιο ειδικούς υποδοχείς. Η RPTPβ/ζ, η εκκρινόμενη μορφή της (φωσφακάνη), αλλά και η κινάση ALK, έχει αναφερθεί ότι μπορούν να δεσμεύουν τη HARP και να συμμετέχουν στη μεταγωγή του σήματός της. Ο κύριος σκοπός αυτής της διατριβής είναι η μελέτη της δομής και της δράσης της HARP, χρησιμοποιώντας μικρότερα τμήματα της HARP τα οποία τα έχουμε βιοτινυλιώσει για να διερευνήσουμε τις περαιτέρω δράσεις της, τους διάφορους μηχανισμούς και με ποιους υποδοχείς αλληλεπιδρά. Τα βιοτινυλιωμένα-σεσημασμένα πεπτίδια αποτελούν χρήσιμα εργαλεία για την βιοτεχνολογία, σε διάφορες εφαρμογές στερεής φάσης ανοσολογικών δοκιμών καθώς και τον εντοπισμό των υποδοχέων. Τα συνθετικά πεπτίδια της HARP συντέθηκαν με μεθοδολογία Fmoc/t-Bu σε στερεά φάση χρησιμοποιώντας ως στερεό υπόστρωμα τη 2-χλωροτρίτυλ-ρητίνη για την παραλαβή μιας ελεύθερης αμινοομάδας (ΝΗ2) και C-τελικού καρβοξυλικού οξέος αντίστοιχα. Για την παραγωγή βιοτινυλιωμένων πεπτιδίων, περιλαμβάνει τον σχηματισμό ενός δεσμού: της ελεύθερη αμινοομάδα με την βιοτίνη στην διάρκεια της πεπτιδικής σύνθεσης στερεής φάσης. Επειδή η βιοτίνη έχει μικρή διαλυτότητα στα διαλύματα της πεπτιδικής σύνθεσης χρησιμοποιούμε τους προσχηματισμένους ενεργοποιημένους εστέρες όπως την βιοτίνη- ONp (biotin p-nitrophenyl ester ). Τα συνθετικά πεπτίδια P(13-39) και Ρ (65-97), που αντιστοιχούσαν στην Ν-CTR-I και C-TSR-I περιοχή της HARP μελετήθηκαν ως προς την επίδρασή τους στον πολλαπλασιασμό και στην μετανάστευση των καρκινικών κυττάρων του προστάτη (PC3). / Heparin affin regulatory peptide (HARP) is an 18 kDa growth factor that has a high affinity for heparin. HARP is highly conserved among species and shares 50% homology with Midkine and RI-HBP. The above proteins constitute a relatively new family of growth factors with high affinity for heparin. HARP has been originally purified from perinatal rat and bovine brain as a molecule that induces neurite outgrowth. HARP is also expressed in uterus, cartilage and bone extracts. Several reports have established a strong correlation between HARP expression and tumour growth and angiogenesis. HARP has been reported to be mitogenic for different types of endothelial cells and angiogenic in vivo and in vitro. HARP exerts its biological activity through interactions with cell surface proteoglycans, such as N-syndecan, or binding to more specific cell surface receptors. Receptor-type protein tyrosine-phosphatase β/ζ (RPTPβ/ζ) and its secreted variant phosphacan, as well as ALK are implicated in HARP signalling. The main target of this study is to investigate the structure and the biological activities of HARP, using smaller fragments of HARP which are biotinylated in order to study its further activities, the mechanisms and the interactions with the receptor. Biotin-labelled peptides are extremely useful tools for biochemistry, with applications in solid-phase immunoassays, affinity purification and receptor localization. The synthetic peptides of HARP were synthesized by Fmoc/t-Bu solid phase methodology utilizing a 2-chlorotrityl-chloride resin to provide a free NH2- amino-group and carboxyl acid, respectively. The simplest approach for the production of biotin-labelled peptides involves capping of a resin-bound free amino group with biotin, prior to cleavage of the peptide from the resin and side-chain deprotection. Because of the poor solubility of biotin in peptide synthesis solvents, biotin is most frequently introduced using a pre-formed active ester, such as biotin p-nitrophenyl ester. The synthetic peptides, P(13-39) and P(65-97) with amino acids sequence corresponding to the N- and C-TSR-I domains of HARP, were studied for their impact to the proliferation and chemotactic (migration) on prostate cancer cells (PC3).

Η HARP (Heparin Affin Regulatory Peptide) είναι ένας αυξητικός παράγοντας ο οποίος εμφανίζει πλειάδα βιολογικών δράσεων εμπλεκόμενος στη διαφοροποίηση, τον πολλαπλασιασμό και τη μετανάστευση πολλών τύπων κυττάρων, καθώς και στην αγγειογένεση και την ανάπτυξη όγκων. Η HARP έχει χρονοειδικό και ιστοειδικό πρότυπο έκφρασης, υπερεκφράζεται όμως σε καρκινικές κυτταρικές σειρές, σε ανθρώπινους καρκινικούς όγκους και βρίσκεται σε υψηλή συγκέντρωση στον ορό του αίματος ασθενών με διάφορες μορφές καρκίνου. Η HARP ασκεί τις βιολογικές της δράσεις μετά από δέσμευση στους διαμεμβρανικούς υποδοχείς, SDC3, ALK και RPTPβ/ζ. Οι βιολογικές της δράσεις προσδιορίζονται από τη συνισταμένη των δράσεων που έχει κάθε υποδοχέας της, αντικατοπτρίζοντας τον περίπλοκο μηχανισμό δράσης της. Στη συγκεκριμένη εργασία μελετήσαμε τον τρόπο με τον οποίο η SDC3 διαμεσολαβεί τις βιολογικές δράσεις της HARP σε κύτταρα DU145 και PC3, κυτταρικές σειρές από καρκίνο ανθρώπινου προστάτη. Χρησιμοποιώντας την RNAi τεχνολογία, διαμολύναμε παροδικά τα κύτταρα με siRNA ειδικά σχεδιασμένο έναντι της SDC3, μειώνοντας τα επίπεδα έκφρασης της. Καλλιεργήσαμε κύτταρα, φυσιολογικά και μετασχηματισμένα, επιδράσαμε εξωγενώς με HARP και τα αποτελέσματα έδειξαν ότι και στις δύο καρκινικές κυτταρικές σειρές, η SDC3 είναι θετικός ρυθμιστής της επαγόμενης από τη HARP κυτταρικής προσκόλλησης και μετανάστευσης. Παράλληλα, μελετήσαμε την ενεργοποίηση μορίων που εμπλέκονται στο μονοπάτι μεταγωγής σήματος της SDC3 όπως της Src, Fak, Akt, Pten και Erk1/2. Τα αποτελέσματα έδειξαν ότι η ενεργοποίησή της SDC3 από τη HARP οδηγεί στην αύξηση των επιπέδων των pSrc, pFak, pAkt και p Erk1/2, ενώ, βρέθηκε ότι μειώνεται η φωσφορυλίωση της Pten. Συμπερασματικά, στην παρούσα εργασία μελετήθηκε η συμμετοχή της συνδεκάνης 3 στις βιολογικές δράσεις της HARP, καθώς και μόρια του σηματοδοτικού μονοπατιού μεταγωγής σήματος του υποδοχέα αυτού. Βρέθηκε ότι η HARP προσδενόμενη στον υποδοχέα αυτό, επάγει την προσκόλληση και μετανάστευση των κυττάρων, δράσεις που συνδέονται με την ανάπτυξη όγκων και τη μετάσταση καρκινικών κυττάρων. / HARP (Heparin Affin Regulatory Peptide), also known as Pleiotrophin, is a growth factor involved in several biological actions such as induction of cellular proliferation, migration and angiogenesis. Elevated concentrations of this growth factor are found in many tumours, as well as in the plasma of patients with different types of cancer. HARP exerts its actions after binding to the transmembrane receptors RPTP β/ζ, ALK and N-Syndecan (SDC3). In the present work, we studied the role of the transmembrane receptor SDC3 in the biological actions of HARP. We used DU145 and PC3 transiently transfected with specific siRNA to downregulate the accumulation of SDC3. Our results show that HARP binds to SDC3 and induces the cell adhesion and migration of DU145 and PC3 cells. We also studied the signal transduction through SDC3 receptor and the activation of signaling molecules such as Src, Fak, Akt, Pten and Erk 1/2. Our results revealed that HARP induces the phosphorylation of Src kinase, Fak and Erk1/2 after binding to SDC3 in both DU145 and PC3 cells. Also, HARP increases Akt signaling cascade in PC3 cells, while it suppresses the signaling cascade induced by PTEN in DU145 cells. Consequently, HARP interaction with SDC3, results in the activation of SDC3, which in turn triggers a signal transduction pathway that leads to specific biological cell responses activates other cytoplasmic effectors. Therefore, there starts a signaling cascade that targets specific genes and cell response. In conclusion, our results indicate that SDC3 contributes, as a positive regulator, to HARP-dependent cell adhesion and migration in both DU145 and PC3 cells.

Articular cartilage plays a vital role in load-bearing joints, providing an almost frictionless surface to articulating bones. However, the avascular nature and low cell density of the tissue means that following injury, there is limited potential for regeneration and repair. With the ageing population, the prevalence and economic burden associated with osteoarthritis (OA) is increasing rapidly, but as of yet there are no fully effective ways to treat the condition. Research into novel therapies has therefore become a popular avenue of investigation, and human mesenchymal stem/stromal cells (hMSCs) have been highlighted as particularly promising targets. However, current, methods for inducing the chondrogenic differentiation of hMSCs, which typically employ the use of transforming growth factor beta 1 or 3 (TGFβ1/3), result in the production of hypertrophic rather than hyaline tissue, hampering translational progress. Growth differentiation factor 5 (GDF5) belongs to the TGFβ superfamily of proteins and is vital for skeletal formation, however its use in cartilage tissue engineering (TE) strategies has been somewhat neglected. Here we demonstrate that GDF5 significantly increases aggrecan gene expression (a marker of articular cartilage), without affecting collagen type X expression (a marker of chondrocyte hypertrophy), in chondrocyte pellet cultures derived from hMSCs, making it a promising target for the formation of permanent articular cartilage. The therapeutic application of growth factors is, at present, limited due to their expense, susceptibility to proteolytic degradation, and rapid clearance, leading to large quantities being required to get anywhere near the desired outcome. The highly sulfated glycosaminoglycan (GAG), heparin, is already extensively used in the clinic as an anticoagulant, and is also able to bind and potentiate the activity of a wide range of growth factors. As such, researchers are now using it to enhance stem cell expansion/ differentiation protocols, as well as to improve the delivery/ activity of growth factors in TE strategies. Here, we identify GDF5 as a novel heparin/heparan sulfate (HS)-binding protein, and show that endogenous HS proteoglycans (HSPGs) are vital for localizing GDF5 to the cell surface, but are not required for its signalling activity. Importantly, we report that clinically relevant doses of heparin (≥ 10 nM), but not equivalent concentrations of HS, inhibit GDF5’s biological activity, in both hMSC-derived chondrocyte pellet cultures, and in the skeletal cell line ATDC5. We demonstrate that these inhibitory effects are due to heparin (but not HS) inhibiting both GDF5 binding to endogenous HSPGs and GDF5-induced induction of Smad 1/5/8 signalling. This study may therefore explain the variable (and disappointing) results seen with heparin-loaded biomaterials for skeletal TE, and the adverse skeletal effects, such as osteoporosis, that have been reported in the clinic following long-term heparin treatment. Together, our results caution the use of heparin in the clinic and in TE applications, and prompt the transition to using more specific GAGs (e.g. HS derivatives or synthetics), with better-defined structures and fewer off-target effects, if optimal therapy is to be achieved. In the case of GDF5, we have used a variety of developed techniques to begin uncovering important structural and functional information regarding the HS-GDF5 interaction, which are hoped to ultimately pave the way towards achieving this aim. Although further analysis is necessary, our data indicate that relatively long HS sequences are required for binding, and that both ionic and non-ionic interactions play a role in the interaction. In addition we suggest that low- rather than high-affinity HS variants may be key to potentiating the activity of this growth factor.

O objeto deste estudo são os eventos hemorrágicos em pacientes críticos que utilizam infusão contínua de heparina sódica. Tem como objetivo geral propor cuidados de enfermagem para pacientes que recebem infusão contínua de heparina, a fim de aumentar a segurança do paciente e reduzir a ocorrência de hemorragia, com base nos fatores de risco. Esta pesquisa procura contribuir com a farmacovigilância da heparina e com a qualidade da assistência de enfermagem. Trata-se de um estudo de coorte retrospectivo, com análise em prontuário, desenvolvido em unidade intensiva e semi-intensiva de um hospital público do Rio de Janeiro. Foram investigados 867 prontuários de 2010 a 2011, encontrando-se uma população de 79 pacientes que fizeram uso de heparina sódica em infusão contínua. As variáveis do estudo foram submetidas a tratamentos estatísticos não paramétricos e a medidas de associação. Os resultados apontam entre os pacientes três diagnósticos: fibrilação atrial, trombose venosa profunda e síndrome coronariana; percebe-se ainda predomínio do sexo feminino (58,23%) e de idosos (md=65 anos). A taxa de eventos hemorrágicos foi de 21,52% e se mostrou mais elevada quando comparada a outros estudos. Evidencia-se que pacientes com TTPa maior do que 100s tem um risco 9,29 vezes maior de apresentar eventos hemorrágicos. Todos os fatores de risco idade maior do que sessenta anos, hipertensão arterial sistêmica, TTPa maior do que 100s, uso prévio de anticoagulante e insuficiência renal apresentam associação positiva com a presença de evento hemorrágico. Entre os pacientes com eventos hemorrágicos, 94,16% apresentam um ou mais fatores de risco para sangramento. Os eventos hemorrágicos foram identificados na pele (47,37%), em sítio de punção, nas vias aéreas, no sistema geniturinário (15,79%) e no sistema gastrointestinal (10,53%). A maioria (55%) dos eventos hemorrágicos foi classificada com tipo 2 de BARC. Na associação entre o dispositivo invasivo utilizado e o tipo de sangramento, 100% dos pacientes com sangramento de via aérea ou do sistema gastrointestinal utilizavam sonda nasoentérica. Paciente com cateter vesical de demora (CVD) tem sete vezes mais risco de hematúria quando comparados com pacientes sem CVD; já pacientes com acesso venoso periférico tem menos risco de sangramento de sítio de punção quando comparados ao pacientes com acesso venoso central (RR= 0,74; 1,29). Essas associações norteiam a assistência de enfermagem e sugerem que o enfermeiro seja cauteloso ao realizar esses procedimentos nos pacientes com heparina sódica. Frente às variações no TTPa dosado, analisou-se o seguimento do protocolo e detectou-se que, nos pacientes com eventos hemorrágicos, a taxa de erro no ajuste da infusão foi maior (76,24%) quando comparada com os pacientes sem eventos hemorrágicos (39,05%). Ao se associar a taxa de erro da infusão com a presença de evento hemorrágico, evidencia-se que, quando a heparina não é ajustada segundo o protocolo, aumenta-se em 3,3 vezes o risco de evento hemorrágico. Portanto, para garantir o uso seguro na infusão de heparina, descrevem-se alguns cuidados específicos de enfermagem baseados nos fatores de risco e na indicação clínica de cada paciente. / The objective of this study are hemorrhagic events in critical patients using continuous infusion of sodium heparin. It aims to propose general nursing care for patients who receive continuous infusion of heparin, in order to increase patient safety and reduce the occurrence of bleeding, based on risk factors. This research seeks to contribute to the pharmacovigilance of heparin and with the quality of nursing care. It is a retrospective cohort study, with chart analysis, developed in intensive and semi-intensive unit of a public hospital in Rio de Janeiro. 867 records of 2010 to 2011 were investigated and a population of 79 patients who made use of sodium heparin in continuous infusion were found. Study variables were subjected to nonparametric statistical treatments and measures of association. The results among patients show three diagnoses: atrial fibrillation, deep vein thrombosis and coronary syndrome; it is also observed female predominance (58.23%) and elderly (average = 65 years old). The rate of hemorrhagic events was 21.52% and was higher when compared to other studies. It was evidenced that patients with APTT (Activated partial thromboplastin time) greater than 100s has a risk of 9.29 times bigger to hemorrhagic events. All risk factors age greater than 60 years old, hypertension, APTT greater than 100s, previous use of anticoagulant and renal insufficiency present positive association with the presence of hemorrhagic event. Among patients with hemorrhagic events, 94.16% have one or more risk factors for bleeding. Bleeding events were identified in skin (47.37%) in puncture site, in the airways, the genitourinary system (15.79%) and in the gastrointestinal system (10.53%). Most of them (55%) of hemorrhagic events was classified with 2 type of BARC. In the association between the invasive device used and the type of bleeding, 100% of patients with bleeding of airways or of the gastrointestinal system using nasoenteric probe. Patient with bladder catheter indwelling (CVD) has seven times more risk of hematuria when compared with patients without CVD; patients with peripheral venous access already has less risk of bleeding from the puncture site when compared to patients with central venous access (RR = 0.74; 1.29). These associations guide nursing care and suggest the nurse to be cautious when performing these procedures in patients with sodium heparin. In variations in dosed APTT, the follow-up Protocol was analyzed and detected that, in patients with hemorrhagic events, the error rate in adjusting the infusion was larger (76.24%) when compared to patients without hemorrhagic events (39.05%). When associating the error rate of infusion with the presence of hemorrhagic event, it shows that when heparin is not adjusted according to the Protocol, increased 3.3 times the risk of hemorrhagic event. Therefore, to ensure safe use on heparin infusion, some specific nursing cares are decribed based on risk factors and on clinical indication of each patient.

Made available in DSpace on 2014-12-17T14:03:38Z (GMT). No. of bitstreams: 1 DayseSA_DISSERT.pdf: 846190 bytes, checksum: eef38ab14de25ca6c9fac44eeac25ae1 (MD5) Previous issue date: 2011-08-05 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / In the last years, heparin has become target of many studies related to inflammation due its ability of biding to proteins involved on immune response. Recently, it was demonstrated, at our laboratory, using a thIoglycollate-induced peritonitis model, heparin s capacity of reduce cellular influx into the peritoneal cavity, 3 hours after the inflammatory stimulus. Once neutrophilic infiltration is highest around 8 hours after the inflammatory stimulus, at the present work, using the same peritonitis model, it was assessed heparin s ability of keeping the interference on leukocyte infiltration, 8 hours after inflammation induction. Moreover, using cellular differential count, it was evaluated how the cellular populations involved in the inflammatory process would be affected by the treatment. Eight hours after the inflammatory stimulus, only heparin dosage of 1 &#956;g/Kg was able to reduce the cellular influx to peritoneum, 62.8% of reduction when compared to positive control (p < 0.001). Furthermore, heparin dosage of 15 &#956;g/Kg presented a pro-inflammatory effect in whole blood verified by the increase of 60.9% (p < 0.001) and 117.8% (p < 0.001) on neutrophils and monocytes proportion, respectively, when compared to positive control. In addition, this dosage also presented a neutrophilic proportion on peritoneal fluid 27.3% higher than positive control (p < 0.05). This duality between anti- and pro-inflammatory effects at different times corroborates studies that attribute a pleiotropic immunomodulator role to heparin. / Nos ?ltimos anos, a heparina vem sendo alvo de v?rios estudos relacionados ? inflama??o, devido sua capacidade de se ligar a diversas prote?nas envolvidas com a resposta imune. Recentemente, em nosso laborat?rio, foi demonstrada, em modelo de peritonite induzida por tioglicolato, a capacidade da heparina em reduzir o influxo celular ? cavidade peritoneal, 3 horas ap?s o est?mulo inflamat?rio. No presente trabalho, utilizando o mesmo modelo, foi avaliada a capacidade da heparina em interferir na infiltra??o leucocit?ria ap?s 8 horas da indu??o da inflama??o, momento em que a infiltra??o neutrof?lica ? m?xima. Utilizando contagem diferencial celular, avaliou-se a influ?ncia da heparina sobre as popula??es celulares envolvidas na inflama??o. Oito horas ap?s o est?mulo inflamat?rio, apenas a dosagem de heparina de 1 &#956;g/Kg, manteve a capacidade de interferir na migra??o leucocit?ria, apresentando 62,8% de diminui??o (p < 0,001) no influxo celular, quando comparada ao controle positivo. Enquanto para a dosagem de heparina de 15 &#956;g/Kg foi observado efeito pr?-inflamat?rio no sangue total verificado pelos aumentos de 60,9% (p < 0,001) e 117,8% (p < 0,001) na propor??o de neutr?filos e mon?citos, respectivamente, em compara??o ao controle positivo. Al?m disso, essa dosagem tamb?m apresentou propor??o neutrof?lica no l?quido peritoneal 27,3% maior que o controle positivo (p < 0,05). Essa dualidade entre efeitos anti- e pr?-inflamat?rio nos diferentes tempos e doses analisados corroboram dados da literatura que atribuem ? heparina um papel como imunomodulador pleiotr?pico.

Orientador: Carlos Alberto Hussni / Banca: Hamilton Almeida Rollo / Banca: Regina Kiomi Takahira / Resumo: A tromboflebite jugular é uma das principais doenças vasculares em eqüinos, podendo levar a edema de cabeça, diminuição do desempenho atlético e até causar o óbito. A heparina é um dos medicamentos utilizados para o tratamento dessa enfermidade com o objetivo de minimizar a evolução do trombo. O objetivo desse experimento foi avaliar, por exames clínicos, laboratoriais, ultra-sonográficos e venográficos as alterações provocadas pela administração de heparina no tratamento da tromboflebite experimental em eqüinos. Dez eqüinos foram submetidos à indução de tromboflebite jugular unilateral. Os animais foram divididos em grupo controle e grupo tratado, composto por cinco animais que receberam durante 10 dias a administração de heparina. Previamente à indução da tromboflebite e diariamente após o estabelecimento da mesma foram realizados exames clínicos e ultra-sonográficos até o 18º dia. Os exames laboratoriais tempo de tromboplastina parcial ativada (TTPA), tempo de protrombina (TP) e contagem total de plaquetas foram determinados no dia anterior à indução da tromboflebite, durante o período de indução e mensurados diariamente, até o 12º dia pós-indução. Hemogramas foram realizados no período pré-indução, imediatamente após a cirurgia para a indução do trombo e no 3°, 7° e 10° dia após a tromboflebite induzida. Medidas diárias do comprimento dos trombos foram submetidas à comparação estatística pelo método de análise de variância. Venografias foram realizadas nos momentos pré-indução, imediatamente à indução, após a indução e a cada seis dias até o 18° dia. No grupo controle, os trombos cresceram até o 9° dia pós-indução, quando foi possível observar diminuição do comprimento dos trombos e aumento de fluxo lateral. No grupo tratado com heparina observou-se a interrupção no crescimento do trombo após ...(Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Jugular thrombophlebitis is a major vascular disease in horses which can lead to head edema, decreased athletic performance and even death. Heparin is used for the treatment of this disease in order to minimize the development of thrombus. The aim of this experiment was to evaluate clinical, laboratory, ultrasound and venographic changes caused by administration of heparin in the treatment of experimental thrombophlebitis in horses. Ten horses were subjected to induction of unilateral jugular thrombophlebitis. The animals were divided into control group and the treated group, composed of five animals each that received during 10 days the administration of heparin. Prior to the induction of thrombophlebitis and daily after the establishment of the same physical examinations and ultrasound were performed until the 18th day. The laboratory tests activated partial thromboplastin (APTT), prothrombin time (PT) and total count platelet were determined the day before the induction of thrombophlebitis, during the induction period and measured daily until the 12th day post-induction. Red blood cells counts were performed in the pre-induction, immediately after surgery for the induction of thrombus in the 3rd, 7th and 10th day after induced thrombophlebitis. Daily measurements of the length of the thrombi were subjected to statistical comparison with analysis of variance. Venographs were performed in the pre-induction, immediately before induction, after induction and every six days until the 18th day. In the control group, the thrombi to grow until the 9th day post-induction, when we observed a shortening of the thrombi and increased lateral flow. In the group treated with heparin there was a break in the growth of the thrombus after the start of administration of the drug and decrease in length of the thrombus with the appearance of flow down the sides of the cranial portion of the thrombus ...(Complete abstract click electronic access below) / Mestre

O objeto deste estudo são os eventos hemorrágicos em pacientes críticos que utilizam infusão contínua de heparina sódica. Tem como objetivo geral propor cuidados de enfermagem para pacientes que recebem infusão contínua de heparina, a fim de aumentar a segurança do paciente e reduzir a ocorrência de hemorragia, com base nos fatores de risco. Esta pesquisa procura contribuir com a farmacovigilância da heparina e com a qualidade da assistência de enfermagem. Trata-se de um estudo de coorte retrospectivo, com análise em prontuário, desenvolvido em unidade intensiva e semi-intensiva de um hospital público do Rio de Janeiro. Foram investigados 867 prontuários de 2010 a 2011, encontrando-se uma população de 79 pacientes que fizeram uso de heparina sódica em infusão contínua. As variáveis do estudo foram submetidas a tratamentos estatísticos não paramétricos e a medidas de associação. Os resultados apontam entre os pacientes três diagnósticos: fibrilação atrial, trombose venosa profunda e síndrome coronariana; percebe-se ainda predomínio do sexo feminino (58,23%) e de idosos (md=65 anos). A taxa de eventos hemorrágicos foi de 21,52% e se mostrou mais elevada quando comparada a outros estudos. Evidencia-se que pacientes com TTPa maior do que 100s tem um risco 9,29 vezes maior de apresentar eventos hemorrágicos. Todos os fatores de risco idade maior do que sessenta anos, hipertensão arterial sistêmica, TTPa maior do que 100s, uso prévio de anticoagulante e insuficiência renal apresentam associação positiva com a presença de evento hemorrágico. Entre os pacientes com eventos hemorrágicos, 94,16% apresentam um ou mais fatores de risco para sangramento. Os eventos hemorrágicos foram identificados na pele (47,37%), em sítio de punção, nas vias aéreas, no sistema geniturinário (15,79%) e no sistema gastrointestinal (10,53%). A maioria (55%) dos eventos hemorrágicos foi classificada com tipo 2 de BARC. Na associação entre o dispositivo invasivo utilizado e o tipo de sangramento, 100% dos pacientes com sangramento de via aérea ou do sistema gastrointestinal utilizavam sonda nasoentérica. Paciente com cateter vesical de demora (CVD) tem sete vezes mais risco de hematúria quando comparados com pacientes sem CVD; já pacientes com acesso venoso periférico tem menos risco de sangramento de sítio de punção quando comparados ao pacientes com acesso venoso central (RR= 0,74; 1,29). Essas associações norteiam a assistência de enfermagem e sugerem que o enfermeiro seja cauteloso ao realizar esses procedimentos nos pacientes com heparina sódica. Frente às variações no TTPa dosado, analisou-se o seguimento do protocolo e detectou-se que, nos pacientes com eventos hemorrágicos, a taxa de erro no ajuste da infusão foi maior (76,24%) quando comparada com os pacientes sem eventos hemorrágicos (39,05%). Ao se associar a taxa de erro da infusão com a presença de evento hemorrágico, evidencia-se que, quando a heparina não é ajustada segundo o protocolo, aumenta-se em 3,3 vezes o risco de evento hemorrágico. Portanto, para garantir o uso seguro na infusão de heparina, descrevem-se alguns cuidados específicos de enfermagem baseados nos fatores de risco e na indicação clínica de cada paciente. / The objective of this study are hemorrhagic events in critical patients using continuous infusion of sodium heparin. It aims to propose general nursing care for patients who receive continuous infusion of heparin, in order to increase patient safety and reduce the occurrence of bleeding, based on risk factors. This research seeks to contribute to the pharmacovigilance of heparin and with the quality of nursing care. It is a retrospective cohort study, with chart analysis, developed in intensive and semi-intensive unit of a public hospital in Rio de Janeiro. 867 records of 2010 to 2011 were investigated and a population of 79 patients who made use of sodium heparin in continuous infusion were found. Study variables were subjected to nonparametric statistical treatments and measures of association. The results among patients show three diagnoses: atrial fibrillation, deep vein thrombosis and coronary syndrome; it is also observed female predominance (58.23%) and elderly (average = 65 years old). The rate of hemorrhagic events was 21.52% and was higher when compared to other studies. It was evidenced that patients with APTT (Activated partial thromboplastin time) greater than 100s has a risk of 9.29 times bigger to hemorrhagic events. All risk factors age greater than 60 years old, hypertension, APTT greater than 100s, previous use of anticoagulant and renal insufficiency present positive association with the presence of hemorrhagic event. Among patients with hemorrhagic events, 94.16% have one or more risk factors for bleeding. Bleeding events were identified in skin (47.37%) in puncture site, in the airways, the genitourinary system (15.79%) and in the gastrointestinal system (10.53%). Most of them (55%) of hemorrhagic events was classified with 2 type of BARC. In the association between the invasive device used and the type of bleeding, 100% of patients with bleeding of airways or of the gastrointestinal system using nasoenteric probe. Patient with bladder catheter indwelling (CVD) has seven times more risk of hematuria when compared with patients without CVD; patients with peripheral venous access already has less risk of bleeding from the puncture site when compared to patients with central venous access (RR = 0.74; 1.29). These associations guide nursing care and suggest the nurse to be cautious when performing these procedures in patients with sodium heparin. In variations in dosed APTT, the follow-up Protocol was analyzed and detected that, in patients with hemorrhagic events, the error rate in adjusting the infusion was larger (76.24%) when compared to patients without hemorrhagic events (39.05%). When associating the error rate of infusion with the presence of hemorrhagic event, it shows that when heparin is not adjusted according to the Protocol, increased 3.3 times the risk of hemorrhagic event. Therefore, to ensure safe use on heparin infusion, some specific nursing cares are decribed based on risk factors and on clinical indication of each patient.

O uso do cateter venoso periférico intermitente em pacientes hospitalizados pode lhes trazer grandes benefícios, proporcionando maior liberdade com a higiene pessoal, alimentação, deambulação, além melhorias para a equipe de enfermagem com redução das rotinas diárias. No entanto, obstrução do cateter é algo que vem na contramão de uma assistência adequada, causando desconforto para o paciente e aumento do trabalho de enfermagem. Para evitar tal evento e garantir a perviedade do cateter intravascular, deve-se realizar o flushing com solução salina ou solução de heparina, após a administração de medicamentos ou em horários programados de acordo com o protocolo interno das instituições de saúde. Nesse contexto, surge o questionamento de qual é a melhor solução para manter a perviedade do cateter venoso periférico intermitente. Este estudo trata-se de uma revisão sistemática da literatura, com o objetivo de avaliar a efetividade do uso da solução salina comparada à solução de heparina na manutenção da perviedade dos cateteres venosos periféricos. Foi realizada a busca nas bases de dados eletrônicas Cinahl, Cochrane Central, Embase, Lilacs, Pubmed e Web of Science. Das 372 referências identificadas, 12 estudos foram selecionados após a análise de título e resumo para serem lidos na íntegra, sendo que 04 constituíram a amostra final. A extração e análise dos dados foram realizadas por dois revisores independentes e as divergências entre eles foram solucionadas por um terceiro revisor. Os 04 estudos foram publicados no período de 1995 a 2015. Ao todo, foram investigados 975 dispositivos endovenosos mantidos com uma das soluções estudadas em 420 participantes. Os estudos testaram a solução de heparina em dosagens que variaram de 10UI/mL a 100UI/mL versus a solução salina e investigaram as complicações relacionadas ao cateter venoso periférico em uso intermitente, sendo uma delas a obstrução do cateter a qual denominamos como ausência e/ou diminuição da perviedade do cateter venoso, que pode ser evitada com o flushing de forma adequada. As intervenções foram testadas em diferentes populações e não foram concordantes em suas conclusões; dois estudos recomendaram a solução de heparina, um estudo, a solução salina, e um recomenda o uso das duas soluções. Os quatro estudos discutem que as duas soluções são seguras e eficazes para a manutenção da perviedade do cateter venoso periférico. Porém, os autores afirmam que o tamanho da amostra não é suficiente para generalizar os resultados para todos os pacientes hospitalizados. Assim, essa revisão sistemática sugere a necessidade de que novos ensaios clínicos randomizados sejam realizados e que as instituições de saúde devem fomentar as investigações clínicas para validar uma assistência de enfermagem alicerçada em práticas seguras e de qualidade / The using of intermittent peripheral venous catheter in a hospitalized patient can bring to them important benefits, providing more freedom concerning to the personal hygiene, feeding, ambulation, and a lot of another improvements for the nursing team, reducing the daily routines. However, catheter obstruction it is something that disturb an appropriate assistence, provoking discomfort to the patient and increasing the need of job of the nursing team. To avoid this fact and to ensure the patency of the intervascular catheter, must to do the flushing process with saline solution or hepamin, after the application of drugs or in pre established time according to the protocol of procedure of the health institutions. In this regard, it emerges the questioning about what is the best solution to keep the patency of the intermittent peripheral venous catheter. This research is a systematic review about the litetature on this subject, and it aims to evaluate the effectiveness of the using of the saline solution in comparison to the heparin in the maintenance of the patency of the peripheral venous catheter. It has been done a research in the electronic databases Cinahl, Cochrane Central, Embase, Lilacs, Pubmed and Web of Science. An amount of 372 references was identified, but just 12 research were selected after the analysis of their titles and abstracts to be fully read. In the end, 04 of them were picked as definitive samples.The data collect and the anlysis were done by two independent reviewers, and the divergences between them were solved by a third reviewer. The 04 research were publicized in the 1995 to 2015 period. Altogheter, were searched 975 intravenous devices, keeping them with one of the studied solutions in 420 patients. Th estudies tried the heparin in dosages that vary from 10UI/mL to 100UI/mL versus the saline solution and investigated the complications related to the peripheral venous catheter in a intermittent using. One of these complications was the catheter obstruction, wich were designated as lack or the decrease of the patency of the venous catheter, that can be avoided with an appropriate flushing process. The interventions were tried in different populations, and they were not concordant in their conclusions. Two research recommended the heparin, one, the saline solution, and one of them recommend to use both. The four research affirm that the heparin and the saline solution are safety and effective for the maintenance of the patency of the peripheral venous catheter. However, the authors say point out that the quantity of the simple is not enough to to generalize the results regarding to all hospitalized patients. Thus, this systematic review suggest the necessity of new randomized clinical testings. In addition, the health institutions must promote clinical research in order to validate a solid nursing assistance based on safety and appropriated practices

Abstract Chronic Chlamydia pneumoniae infections have been associated with cardiovascular diseases (CVD), but the treatment is difficult. Some drugs used for CVD have been found to have an inhibitory effect on the C. trachomatis infection, which is not considered to be associated with CVD. The purpose of this study was to investigate the effects of heparan sulfate-like glycosaminoglycans, COX inhibitors and rapamycin on the C. pneumoniae infection with cell culture methods. Almost any conceivable factors may affect the results of cell cultures. This study showed the complex interaction between temperature, time and medium during the pre-treatment before inoculation. The influences of these factors on the results overlapped and interlaced. The simple washing procedure could enhance the infectivity of C. pneumoniae although it is generally considered to cause the loss of chlamydial EBs and sequentially decrease the chlamydial infectivity. Although the detailed mechanisms were not studied, the results of this study showed that selective COX inhibitors and rapamycin can inhibit the infectivity of C. pneumoniae by inhibiting the growth and maturation, whereas heparan sulfate-like glycosaminoglycans perhaps inhibit the attachment of C. pneumoniae EBs onto the host cells. Recovery and repassage results showed that the growth can be only delayed by selective COX inhibitors, and it can recover to normal level once the drugs were removed. However, rapamycin inhibited the maturation of chlamydial EBs and therefore the infectivity fell down further even when the rapamycin was removed. This study also presented the variations of pathogenicity between different C. pneumoniae strains in vitro. This study is based on in vitro experiments with an acute infection model. Thus, any definite conclusions on the possible antichlamydial effects of the drugs tested in the treatment of cardiovascular diseases which are associated with chronic C. pneumoniae infections cannot be drawn on the basis of this study.

Relative changes are detectable in the blood of end-stage renal disease (ESRD) patients during hemodialysis (HD) treatment using optical spectroscopy. However, the potential impacts of several confounding factors that could affect the detection of these changes have not been evaluated. The objectives of this thesis were to: 1) investigate how the variations and/or changes in acid-base and oxygen parameters during HD treatment can affect the optical signature of whole blood of ESRD patients, 2) to investigate the effect of heparin on the optical properties of whole blood and its impact on our method. Blood samples were drawn from 23 ESRD patients at 5 time points during a 4 hour HD treatment and sent for blood gas and blood spectroscopy analyses. No significant correlations were found between the changes in the blood transmittance spectra and acid-base and oxygen parameters. This indicates that the perturbations in these parameters due to HD procedures do not confound the detection of changes in the blood transmittance spectra of ESRD patients during HD treatment. Additionally, the effect of heparin in modifying the optical properties of whole blood does not confound the detection of changes in the blood of ESRD patients due to HD treatment using whole blood-based optical spectroscopy. ANOVA revealed significant (P<0.05) measurable changes in the blood transmittance spectra of ESRD patients during HD treatment. Significant spectral differences (P<0.05) were found between ESRD patients. The lack of uniform spectral characteristics across patients is