Rol del sistema kallicreína kinina y su interrelación con sistemas vasoactivos durante la preñez / Role of the kallikrein kinin system and its interrelationship with vasoactive systems in pregnancy

La hiperfiltración glomerular y el aumento de la reabsorción de sodio son factores fundamentales para el desarrollo de la unidad feto placentaria. Dichos factores resultan de adaptaciones hemodinámicas y renales en las que participan sistemas vasoactivos. Se pudo demostrar en ratas que la activación del sistema kallicreína kinina (SKK) precede a la instalación de la hiperfiltración glomerular, dado que su inhibición por aprotinina previene el aumento del filtrado glomerular. Además, la inhibición individual o asociada de los efectores específicos del SKK, las prostaglandinas (PGs) y el óxido nítrico (ON), confirman la dependencia del filtrado glomerular del SKK durante la preñez. Encontramos también que el sistema renina angiotensina (SRA) participa en la generación de la hiperfiltración glomerular dado que ésta es afectada por la administración de bloqueantes del SRA. La inhibición máxima sobre el pico de hiperfiltración se obtuvo con el bloqueo de ambos sistemas (SKK y SRA). Además, estrategias para alterar la hiperfiltración glomerular y la reabsorción de sodio de la preñez evidenciaron alteraciones en el desarrollo de la unidad feto placentaria, menor número de crías, mayor cantidad de reabsorciones intrauterinas y retardo en el crecimiento. El tratamiento combinado de inhibidores del SKK asociados a bloqueantes del SRA o de óxido nítrico mostraron los mayores efectos. En consecuencia, demostramos que el SKK juega un rol central en los fenómenos de adaptación que acompañan la preñez normal. La interrelación del SKK con varios sistemas vasoactivos parecería formar una red que participa en las adaptaciones hemodinámicas para un adecuado desarrollo de la gestación y de la unidad feto-placentaria.

Glomerular hyperfiltration and increased sodium reabsorption are key factors for the development of the fetus and placenta in pregnancy. These adjustments result from hemodynamic and renal factors involving vasoactive systems. It was demonstrated in rats that activation of KKS precedes the installation of glomerular hyperfiltration as aprotinin prevents the increase in glomerular filtration. In addition, individual or associated inhibition of specific kallikrein kinin system effectors, prostaglandins (PGs) and nitric oxide (NO), confirm the glomerular filtration rate dependence of KKS during pregnancy. It was also found that the renin-angiotensin system (RAS) contributes to glomerular hyperfiltration as this is affected by the administration of RAS blockers. The peak of hyperfiltration maximum inhibition was obtained by the blockade of both systems (KKS and RAS). In addition, strategies used to alter the glomerular hyperfiltration and increased sodium reabsorption during pregnancy, showed abnormalities in the development of the fetus and placenta, fewer offspring, more fetus resorptions and intrauterine growth retardation. KKS inhibitors associated with RAS or nitric oxide blockers showed the greatest impact. As a consequence, it was demonstrated that KKS plays a central role in the adaptation phenomenom that accompanies normal pregnancy. The interplay of KKS with several vasoactive systems, seem to arrange a network involved in the hemodynamic adaptations to allow the proper development of pregnancy and the fetus and placenta.

Avaliação da aprotinina na redução da resposta inflamatória sistêmica em crianças operadas com circulação extracorpórea / Assessment of aprotinin in the reduction of inflammatory systemic response in children undergoing surgery with cardiopulmonary bypass

OBJETIVO: Avaliar se a aprotinina em altas doses hemostáticas pode reduzir o processo inflamatório após circulação extracorpórea (CEC) em crianças. MÉTODOS: Estudo prospectivo randomizado em crianças de 30 dias a 4 anos de idade, submetidas à correção de cardiopatia congênita acianogênica, com CEC e divididas em dois grupos, um denominado Controle (n=9) e o outro, Aprotinina (n=10). Neste, o fármaco foi administrado antes e durante a CEC. A resposta inflamatória sistêmica e disfunções hemostática e multiorgânicas foram analisadas por marcadores clínicos e bioquímicos. Foram consideradas significantes as diferenças com P<0,05. RESULTADOS: Os grupos foram semelhantes quanto às variáveis demográficas e intra-operatórias, exceto por maior hemodiluição no Grupo Aprotinina. Não houve benefício quanto aos tempos de ventilação pulmonar mecânica, permanência no CTIP e hospitalar, nem quanto ao uso de inotrópicos e função renal. A relação PaO2/FiO2 (pressão parcial de oxigênio arterial/fração inspirada de oxigênio) apresentou queda significativa com 24 h pós-operatório, no Grupo Controle. As perdas sanguíneas foram semelhantes nos dois grupos. No grupo Aprotinina surgiu leucopenia significativa, em CEC, seguida de leucocitose. Fator de necrose tumoral alfa (TNF-α), Interleucinas (IL)-6, IL-8, IL-10, proporção IL-6/IL-10 não apresentaram diferenças marcantes intergrupos. A proporção IL-6/IL-10 PO aumentou no grupo Controle. Não houve complicações com o uso da aprotinina. CONCLUSÃO: Nesta casuística, a Aprotinina em altas doses hemostáticas não minimizou as manifestações clínicas e os marcadores séricos de resposta inflamatória sistêmica.

OBJECTIVE: To evaluate if the hemostatic high-dose aprotinin seems to reduce the inflammatory process after extracorporeal circulation (ECC) in children. METHODS: A prospective randomized study was conducted on children aged 30 days to 4 years submitted to correction of acyanogenic congenital heart disease with ECC and divided into two groups: Control (n=9) and Aprotinin (n=10). In the Aprotinin Group the drug was administered before and during ECC and the systemic inflammatory response and hemostatic and multiorgan dysfunctions were analyzed on the basis of clinical and biochemical markers. Differences were considered to be significant when P<0.05. RESULTS: The groups were similar regarding demographic and intraoperative variables, except for a greater hemodilution in the Aprotinin Group. The drug had no benefit regarding time of mechanical pulmonary ventilation, permanence in the postoperative ICU and length of CONCLUSION: In this series, hemostatic high-dose aprotinin did not minimize the clinical manifestations or serum markers of the inflammatory systemic response.

The Comparison of Parathyroid Hormone Degradation Effect by Various Protease Inhibitors in Blood Specimen / 대한진단검사의학회지

BACKGROUND: The objective of this study was to evaluate the role of proteases on the degradation of parathyroid hormone (PTH) in blood samples. METHODS: Protease inhibitors with specificity against serine proteases (aprotinin), cysteine proteases (E-64), serine and cysteine proteases (leupeptin), metalloproteases (EDTA), or a protease inhibitor cocktail with a broad spectrum of inhibitory activity were added to blood samples. After storage at room temperature (0-48 hr), PTH levels were measured. RESULTS: PTH levels in samples with the protease inhibitor cocktail did not change significantly after 48 hr of storage at room temperature, but the average PTH levels decreased by 40.7% and 20.1%, in samples stored at room temperature and stored at 4degrees C without protease inhibitors, respectively. PTH levels in samples with leupeptin were stable for up to 24 hr. After 48 hr, the mean PTH levels decreased by 17.1%, 16.0%, 26.2%, and 32.1%, with 500 KIU/mL aprotinin, 100 micro mol/L leupeptin, 10 micro mol/L E-64, and 10 micro mol/L EDTA, respectively, in the samples stored at room temperature. CONCLUSIONS: The decrease in PTH levels in blood samples seemed to be due to the degradation of PTH by proteases. Various proteases, including especially serine proteases, would act together to degrade PTH in blood specimen. The PTH degradation may be inhibited in blood specimen with protease inhibitor cocktail.

Purification of elastase-like chymotrypsin from cardamom shoot and Capsule borer [corrected]

An elastase-like chymotrypsin was purified by aprotinin-agarose affinity chromatography from the midgut extract of cardamom shoot and capsule borer, Conogethes punctiferalis. The purified enzyme had a Vmax of 687.6 +/- 22.1 nmole pNA released/min/mg protein, Km of 0.168 +/- 0.012 mM with SAAPLpNA as substrate and gave a single band on SDS-PAGE with a molecular mass of 72.1 kDa. Casein zymogram revealed one clear zone of proteolytic activity, which corresponded to the band obtained with SDS-PAGE indicating that this could be a single-polypeptide enzyme.

Antifibrinolíticos e cirurgia cardíaca com circulação extracorpórea / Antifibrinolytics and cardiac surgery with cardiopulmonary bypass

JUSTIFICATIVA E OBJETIVOS: A cirurgia cardíaca é a especialidade cirúrgica que com mais freqüência está associada a sangramento, coagulopatia e necessidade de derivados de sangue. Os agentes farmacológicos aprotinina, ácido epsilon-aminocapróico e ácido tranexâmico são os mais utilizados para auxiliar na hemostasia dos pacientes submetidos à circulação extracorpórea. O objetivo deste trabalho foi apresentar a fisiopatologia do sangramento em cirurgia cardíaca e a atual situação dos antifibrinolíticos quanto à sua eficácia e complicações quando usados nesses procedimentos, dando maior ênfase ao ácido tranexâmico e à aprotinina. CONTEUDO: São discutidos os mecanismos pelos quais a circulação extracorpórea provoca alteração na hemostasia e de que maneira os antifibrinolíticos agem para diminuir o sangramento e o uso de sangue alogênico em cirurgia cardíaca. É dada ênfase ao problema do tromboembolismo que pode ocorrer com o emprego desses antifibrinolíticos, com revisão da literatura. CONCLUSÃO: A fibrinólise é um dos principais fatores relacionados com o aumento do sangramento em cirurgia cardíaca com circulação extracorpórea. A inibição da fibrinólise, juntamente com a preservação da função plaquetária, é, provavelmente, o mecanismo pelo qual os antifibrinolíticos diminuem o sangramento. O emprego desses fármacos reduz o sangramento em cirurgia cardíaca com circulação extracorpórea num percentual que pode chegar a 50 por cento. Com relação à preocupação com o tromboembolismo, os ácidos tranexâmico e epsilon-aminocapróico são opções que oferecem maior segurança que a aprotinina.

BACKGROUND AND OBJECTIVES: Cardiac surgery is the surgical subspecialty most often associated with bleeding, bleeding disorders, and the need of blood products. Agents such as aprotinin, episilon-aminocaproic acid, and tranexamic acid are frequently used to aid the hemostasis of patients undergoing cardiopulmonary bypass. The objective of this report is to present the physiopathology of bleeding during cardiac surgeries and the current role of antifibrinolytics regarding their efficacy and complications when used in those procedures, with emphasis on tranexamic acid and aprotinin. CONTENTS: The mechanisms of changes in hemostasis caused by cardiopulmonary bypass, how antifibrinolytics decrease bleeding, and the use of alogenic blood in cardiac surgery are discussed. A review of the literature emphasizes the thromboembolism secondary to the use of those antifibrinolytics. CONCLUSION: Fibrinolysis is one of the main factors related with increased bleeding during cardiac surgery with cardiopulmonary bypass. Inhibition of fibrinolysis associated with the preservation of platelet function is, probably, the mechanism by which anti-fibrinolytics decrease bleeding. Those agents reduce bleeding in up to 50 percent in cardiac surgeries with cardiopulmonary bypass. Tranexamic acid and episilon-aminocaproic acid are safer than aprotinin in the prevention of thromboembolism.

JUSTIFICATIVA Y OBJETIVOS: La cirugía cardiaca es la especialidad quirúrgica que más frecuentemente está asociada al sangramiento, cuagulopatía y con necesidad de derivados de sangre. Los agentes farmacológicos aprotinina, ácido epsilon-aminocapróico y el ácido tranexámico son los más utilizados para auxiliar en la hemostasia de los pacientes sometidos a la circulación extracorpórea. El objetivo de este trabajo fue presentar la fisiopatología del sangramiento en cirugía cardiaca y la actual situación de los antifibrinolíticos en cuanto a su eficacia y complicaciones cuando usados en estos procedimientos dando más énfasis al ácido tranexámico y a la aprotinina. CONTENIDO: Son discutidos los mecanismos por los cuales la circulación extracorpórea provoca alteración en la hemostasia y de que manera los antifibrinolíticos actúan para disminuir el sangramiento y el uso de sangre alogénica en cirugía cardiaca. Se le da énfasis al problema del trombo embolismo que puede ocurrir con el uso de esos antifibrinolíticos, con revisión de la literatura. CONCLUSIONES: La fibrinólisis es uno de los principales factores relacionados con el aumento del sangramiento en cirugía cardiaca con circulación extracorpórea. La inhibición de la fibrinólisis, conjuntamente con la preservación de la función plaquetaria es probablemente el mecanismo por el cual los antifibrinolíticos disminuyen el sangramiento. El uso de esos fármacos reduce el sangramiento en cirugía cardiaca con circulación extracorpórea en un porcentaje que puede alcanzar el 50 por ciento. Con relación a la preocupación con el trombo embolismo, el ácido tranexámico y el ácido epsilon-aminocapróico son opciones que ofrecen una mayor seguridad que la aprotinina.

Aprotinin Attenuates the Elevation of Pulmonary Vascular Resistance After Cardiopulmonary Bypass

Pulmonary vascular resistance (PVR) is generally believed to be elevated after cardiopulmonary bypass (CPB) due to whole body inflammation. Aprotinin has an antiinflammatory action, and it was hypothesized that aprotinin would attenuate the PVR increase induced by CPB. Ten mongrel dogs were placed under moderately hypothermic CPB for 2 hr. The experimental animals were divided into a control group (n=5, group I) and an aprotinin group (n=5, group II). In group II, aprotinin was administered during pre-bypass (50,000 KIU/kg) and post-bypass (10,000 KIU/kg) periods. Additional aprotinin (50,000 KIU/kg) was mixed in CPB priming solution. PVRs at pre-bypass and post-bypass 0, 1, 2, 3 hr were calculated, and lung tissue was obtained after the experiment. Post-bypass PVRs were significantly higher than prebypass levels in all animals (n=10, p<0.001). PVR elevation in group II was less than in group I at 3 hr post-bypass (p=0.0047). Water content of the lung was lower in group II (74+/-9.4%) compared to that of group I (83+/-9.5%), but the difference did not reach significance (p=0.076). Pathological examination showed a near normal lung structure in group II, whereas various inflammatory reactions were observed in group I. We concluded that aprotinin may attenuate CPB-induced PVR elevation through its anti-inflammatory effect.

Effect of aprotinin on schistosoma mansoni miracidial penetration into biomphalaria alexandrina

The effect of aprotinin on Schistosoma mansoni miracidial penetration process, by its direct topical application on Biomphalaria alexandrina snails, was studied. The snails were exposed to S. mansoni miracidial suspension which was mixed with aprotinin at concentrations ranging from 0.02 to 20 Kallikrein inactivator units [KIU]/ml of applied solution. Results showed that aprotinin had marked inhibitory effect on miracidial penetration of the snails. The concentration that resulted in 50% inhibition of the miracidial penetration into B. alexandrina snails [LD50] was 2.4 KIU/ml, while [LD96] was 20 KIU/ml of the applied solution. Thus, aprotinin may be one of the important methods in the control of schistosomiasis

Kinetic mechanism of the inhibition of human urinary kallikrein by basic pancreatic trypsin inhibitor

Hydrolysis of D-valvyl-L-leucyl-L-arginine p-nitroanilide (D-Val-Leu-Arg-Nan) at five different concentration (10-20µM) by human urinary kallikrein was studied in the absence and in the presence of increasing concentrations of basic pancreatic trypsin inhibitor (BPTI) (1.35-9.15nM). The data indicate that the inhbition of human urinary kallikrein by BPTI is not a simple competitive inhibition as reported by others, but that it is a competitive inhibition of the parabolic type, with two inhibitor molecules binding to one enzyme molecule, with the formetion of a ternary enzymatic complex. Statistical analysis of the experimental data supports the kinetic model proposed. The calculated values of the constants Ki and Kii were 16.20 nM and 1.10 nM, repectively. It is noteworthy that the Kii < Ki, i.e., the second BPTI molecule binds to the enzyme with a larger affinity suggesting that this second binding site was probably created or positively modulated as a consequence of the binding of the first BPTI molecule