combination effect of hypertonic disease with chronic pancreatitis on the processes maintain homeostasis.

OBJECTIVE: Introduction: Abnormalities comorbidity - a frequent phenomenon in medical practice. This determines the relevance of research processes maintaining homeostasis with a combination of various diseases. The aim of this study was to examine and compare the character of vegetative, antioxidant, kallikrein-kinin system and parameters of endogenous intoxication disorders in the patients with isolated essential hypertension and with combination of hypertonic disease and chronic pancreatitis. PATIENTS AND METHODS: Materials and Methods: Cardiointervalography was used in the research with definition of standard statistical and spectral heart rate variability. Determination of superoxide dismutase, glutathione, catalase, middle molecular peptides, total proteolytic activity of plasma by the hydrolysis of protamine sulfate, prekallikrein, kallikrein, α1 -proteinase inhibitor, α2 -macroglobulin and kininase II was conducted by laboratory methods. RESULTS: Results: Sympathicotonia with the moderate tension of adaptation processes, violation of antioxidant protection, kallikrein-kinin system and displays of endogenous intoxication were found in the patients with isolated hypertension. Reduction of sympathicotonia, reducing total power spectrum, increasing the share of humoral-metabolic effects on heart rate, tendency to asympathicotonia autonomic reactivity, lower levels of superoxide dismutase, glutathione, prekallikrein, α2 -macroglobulin, kininase II, higher levels of catalase, middle molecular peptides, total proteolytic activity of plasma kallikrein were observed upon accession the concomitant chronic pancreatitis. CONCLUSION: Conclusions: The signs of compensatory mechanisms disruption and increased autonomic nervous system imbalance with a decrease in ductility autonomous processes in the load were determined upon accession the concomitant chronic pancreatitis. The combination of pathologies also accompanied by more severe manifestations of endogenous intoxication, significant violations of antioxidant and kallikrein-kinin systems.

von Willebrand Factor and Prekallikrein in Plasma Are Associated With Thrombus Volume in Abdominal Aortic Aneurysms.

OBJECTIVES: Disruption of the endothelial lining may be one of the events linking intraluminal thrombus and abdominal aortic aneurysm growth. In the present study, we examined whether von Willebrand factor activity in plasma, contact proteins of blood coagulation, and inflammatory biomarkers may be associated with intraluminal thrombus volume in search of a biochemical marker of endothelial damage and thrombus size. DESIGN: Prospective study, correlating potential endothelial biomarkers and intraluminal thrombus volume acquired by computed tomography angiography. MATERIALS AND METHODS: Plasma was consecutively obtained from 38 patients with asymptomatic infrarenal abdominal aortic aneurysm. von Willebrand factor activity, thrombin generation time, factor XII, and prekallikrein concentration were measured in plasma on automated and in-house platforms. In total, 8 patients were excluded due to ongoing anticoagulant therapy, renal impairment, or nonappearance, thus leaving 30 patients for further analysis. All patients had computed tomography angiography, and intraluminal volume was quantified off-line by OsiriX 6.5. RESULTS: Median intraluminal thrombus volume was 42.7 mL. Spearman correlation analysis revealed a positive correlation between thrombus volume, von Willebrand factor activity (ρ = 0.56, P = .0013), and prekallikrein concentration in plasma (ρ = 0.54, P = .002). CONCLUSION: von Willebrand factor activity and concentration of prekallikrein may both be of importance regarding the evolution of thrombus in abdominal aortic aneurysm and possible biomarkers for aneurysm growth.

[Prekallikrein (Fletcher Factor) Deficiency and Prolongation of APTT Reaction]. / Präkallikrein-(Fletcher-Faktor-)Defekt als Ursache einer präoperativen APTT-Verlängerung.

BACKGROUND: Prekallikrein (Fletcher factor) is a protein of the contact phase of the blood coagulation system. A deficiency is a very rare event. The clinical meaning is still unknown. CASE REPORT: In a 24-year-old patient of Croatian nationality, who has never suffered from any kind of proneness to hemorrhage, a considerable prolongation of activated partial thromboplastin time (APTT) was diagnosed preoperatively. The prothrombin time (PT) was normal. The patient had to be subjected to horseshoe-kidney surgery. CLOTTING TESTS: A very seldom Fletcher factor (prekallikrein) deficiency type II was detected. The activity was < 1%, but the prekallikrein concentration was normal, comparable to that in normal plasma. THERAPY AND COURSE: Complications were neither noticed during renal surgery nor postoperatively. The patient did not show clinical symptoms. This fact corroborates reports about similar cases published earlier. CONCLUSION: Nevertheless, attention has to be paid to these exceptionally rare symptoms, as in some publications, thromboembolic or even bleeding complications were reported.

Elevated levels of plasma prekallikrein, high molecular weight kininogen and factor XI in coronary heart disease.

Increased levels of hemostatic factors may play a role in the pathogenesis of myocardial infarction by triggering thrombin formation. We measured factor XII (FXII), factor XI (FXI), plasma prekallikrein (PK) and high-molecular-weight kininogen (HK) in 200 patients having survived myocardial infarction for at least 2 months, and in 100 healthy controls. We found significantly elevated levels of FXI clotting activity (FXI:C), HK:C and of the amidolytic activity of PK (PK:Am) among the patients as compared to the controls. Plasma levels of FXI:C, HK:C and PK:Am in the highest quartile were associated with an odds ratio of 1.9 (95% CI: 1.0-3.8), 2.0 (95% CI: 1.0-4.0) and 5.4 (95% CI: 2.6-11.2), respectively, compared to the respective plasma levels in the lowest quartile. After correction for established clinical and laboratory risk factors, the association between PK:Am plasma levels and myocardial infarction remained significant (P=0.0007). Combination of high PK:Am plasma levels and smoking or arterial hypertension, respectively, resulted in a more than additive relative risk for myocardial infarction.

Effect of C1-esterase-inhibitor on capillary leak and inflammatory response syndrome during arterial switch operations in neonates.

OBJECTIVE: To determine if prophylactic administration of C1-esterase-inhibitor would have a beneficial effect on postoperative weight gain and the inflammatory response in neonates undergoing cardiac surgery with cardiopulmonary bypass (CPB). DESIGN: Randomized, double-blinded study. SETTING: University-affiliated heart center. PARTICIPANTS: Twenty-four neonates with transposition of the great arteries. INTERVENTIONS: In group inhibitor (INH) patients (n = 12), 100 IU/kg of C1-esterase-inhibitor (Berinert) was given 30 minutes before CPB. In group placebo (P) patients (n = 12), placebo was administered instead. Interleukin (IL)-6, C3a anaphylatoxin, C1 activity, prekallikrein, Hageman factor, D-dimers, and clinical parameters were measured 6 times perioperatively. MEASUREMENTS AND MAIN RESULTS: All 24 patients had an uneventful clinical course. Mean arterial pressure and pulmonary oxygenation after CPB were superior in group INH patients. The weight gain on postoperative days 1 to 4 was significantly less in group INH patients compared with group P (55 +/- 59 g vs. 340 +/- 121 g, day 1). The concentration of IL-6 (76 +/- 17 pg/mL vs. 262 +/- 95 pg/mL during CPB) was significantly lower in group INH patients compared with group P patients. In contrast, no influence on C3a anaphylatoxin and coagulation factors was found. CONCLUSION: Prophylactic application of C1-esterase-inhibitor in neonates undergoing arterial switch operations produces less inflammatory response compared with placebo. This difference may have contributed to improved clinical parameters, including less weight gain postoperatively.

Removal of IgG from normal plasma and plasma from untreated patients with active Crohn's disease--effect on levels of contact factors.

Protein G columns were used to remove IgG from human plasma, and the effect on levels of factor XII, factor XI and prekallikrein was studied in functional tests. IgG was detected in PAGE immunoblot experiments with Fc-specific antibodies. Removal of the bulk of IgG in a procedure based on a low plasma dilution (1+2.5) allowed the passage of an IgG fraction along with the contact factors. This fraction was found to be present in higher amounts in plasma from patients with Crohn's disease (n=5) than in control plasma (n=12). In a previous study, PAGE immunoblot experiments showed that part of the prekallikrein was removed along with IgG when a higher plasma dilution (1+10.8) was used (Scand J Clin Lab Invest 1999; 59: 55-64). This observation was supported by results in the present work based on parallel assays with the peptide substrates S-2302 and Bz-Pro-Phe-Arg-pNA. The prekallikrein fraction removed was present in a functional state differing from the main part of prekallikrein by yielding kallikrein with a significantly increased activity against the substrate S-2366. This prekallikrein fraction was present in higher amounts in patient plasma than in control plasma. Part of the corresponding amidase activity was blocked by lima bean trypsin inhibitor, suggesting its presence in association with factor XI. The results also indicated that prekallikrein activator activity was connected with this fraction. With the high dilution procedure an extensive removal of IgG from the patient plasma was obtained compared to the control plasma.

Contact activation factors in plasma from women on estrogen replacement therapy after ovariohysterectomy.

The plasma levels of factor XII, prekallikrein, factor XI, and high molecular weight kininogen were studied in women with bilateral oophorectomy and hysterectomy who received hormone replacement therapy with a 2 mg daily dose of estradiol valerate. Also plasminogen activator activity was investigated. The observations made provide support for the assumption that the low doses of estrogen used in hormone replacement therapy do not significantly affect the levels of contact activation or fibrinolytic factors in plasma. Plasma obtained from young, healthy women was used as a standard reference material. Significantly higher levels of factor XII and prekallikrein were registered in functional tests in the ectomized women than in the reference material, an increase not observed in the immunological assays. These observations are discussed in light of recently published data from our laboratory on an increase in the measured level of factor XII obtained upon the removal of IgG before assay. Also a marked increase in urokinase activity was registered in the ectomized women. The high levels of factor XII, prekallikrein, and urokinase, as compared with the reference material, seemed to be age dependent, being also observed in a group of naturally postmenopausal women.

The precursor form of the human kallikrein 2, a kallikrein homologous to prostate-specific antigen, is present in human sera and is increased in prostate cancer and benign prostatic hyperplasia.

Prostate-specific antigen (PSA, hK3) is a diagnostic marker for prostatic cancer but lacks the specificity to sufficiently distinguish between prostatic cancer and benign prostatic hyperplasia (BPH). Human glandular kallikrein 2 (hK2) has been proposed as a potential diagnostic marker for prostate cancer that could complement the current PSA test. Recently we demonstrated that proPSA is present in prostate cancer sera. This study examines the expression of prohK2 in prostate cells and its presence in human sera. Western blot analysis was used to assess prohK2 expression in the human carcinoma cell line, LNCaP. A highly specific and sensitive dual monoclonal immunoassay for prohK2 was developed and used to assess the presence of prohK2 in human sera. prohK2 was detected in the spent media of LNCaP cells. Furthermore, prohK2 was present at immunodetectable concentrations in human sera, and its concentration was increased in prostatic cancer and BPH. These results indicate for the first time that prohK2 is secreted by human prostate cells and is a major component of uncomplexed (free) hK2 in human sera. In addition, prohK2 in human sera is associated with prostate disease and thus may be a useful marker for prostatic cancer and BPH.

[Hemostatic disorders as a complication of autoimmune hemolytic anemia in dogs]. / Hämostasestörungen als Komplikation der autoimmunhämolytischen Anämie beim Hund.

In eleven of 15 investigated dogs suffering from autoimmune haemolytic anaemia an increased concentration of soluble fibrin (p < 0.001) and in all of these 15 dogs an increased concentration of fibrin(ogen) degradation products was observed. The increased turnover indicated by these markers of intravascular coagulation and hyperfibrinolysis was decompensated in approximately the half of the patients which was shown by a decrease of platelet count, activity of different coagulation factors as well as of the inhibitor antithrombin III. The consumption induced decrease of activity was especially related to prekallikrein and high molecular weight kininogen reflecting an activation mainly of the initial intrinsic system by the haemolytic anaemia and was also indicated by the activated partial thromboplastin time which was distinctly prolonged in some cases. The results of this study demonstrate the necessity of an anticoagulant therapy in dogs suffering from autoimmune haemolytic anaemia.

Studies on the components of the contact phase system in patients with acute nonlymphoblastic leukemia

The objective of the present study was to evaluate factors of the plasma kallikrein system in patients with acute nonlymphoblastic leukemia (ANLL), and compare the results to a normal control group. A prospective study was performed in the Tertiary Health Care Institution, Hemocentro, Campinas State University, Campinas, Sao Paulo, Brazil. Thirty-five patients, diagnosed as ANLL between 1988 and 1991, were considered for participation. Eleven patients were not elegible, according to the exclusion criteria: infection/septicemia, previous treatment of blood transfusion. The study was performed with 24 ANLL patients, average age 34 years (16-69 years), 14 men and 10 women. Nineteen healthy volunteers, workers from the Hematology Center, average age 32 years (21-59 years), 11 men and 8 women, were the control group. Plasmatic prekallikrein, C1-inhibitor, alpha 2-macroglobulin, activated partial thromboplastin time, prothrombin time, factor XII, factor XI, factor V and prealbumin were measured. Plasmatic prekallikrein (p=0.02) and prealbumin (p=0.03) were significantly decreased, and prothrombin time increased (p=0.003) in the patient group when compared to the control. Significant correlation (r=0.49, critical value=0.43, p<0.05) between prekallikrein and prealbumin, and between prothrombin time and factor V (r=0.54, critical value=0.44, p<0.05) was demonstrated in the patient group. No correlation was found between parameters analysed and circulant blast count or leukemia subgroups. Statistical analysis was performed by the Willcoxon test. Correlation between the parameters was also verified. These results suggest activation of the contact system or impaired liver synthesis in patients with ANLL, and could contribute to disease complications.

Activation of the contact system by filtration of platelet concentrates with a negatively charged white cell-removal filter and measurement of venous blood bradykinin level in patients who received filtered platelets.

BACKGROUND: Several recent reports have described hypotensive transfusion reactions in patients receiving platelet concentrates (PCs) filtered through white cell-reduction filters. It is well known that a negatively charged surface activates the contact system, consisting of factor XII, prekallikrein, and high-molecular-weight kininogen. STUDY DESIGN AND METHODS: To clarify the mechanisms of these hypotensive reactions, the possibility that white cell-reduction filtration activates the contact system was examined. Venous blood plasma bradykinin levels were also measured in patients receiving PC transfusions through filters. RESULTS: None of the measured values were changed by filtration through a positively charged filter. However, filtration through a negatively charged filter resulted in a decrease in the amounts of prekallikrein and an increase in the amount of bradykinin generated, which indicated the activation of the contact system. The bradykinin level was inversely related to the activity of angiotensin-converting enzyme (ACE) in the PCs and was elevated by addition of an ACE inhibitor. Although the venous blood plasma bradykinin level did not change in two patients with a normal ACE activity during PC transfusion through the negatively charged filter, two patients who had decreased ACE activity, showed a significant increase in bradykinin during the transfusion. CONCLUSION: These results suggest that the generation of a large amount of bradykinin by filtration of PCs through a negatively charged filter might cause hypotensive reactions in patients with decreased ACE activity. The clinical significance of bradykinin generation requires further study.

Cleavage of plasma high molecular weight kininogen in surgical ICU patients.

OBJECTIVE: To characterize kininogens in plasma from surgical patients in the intensive care unit (ICU). DESIGN: Prospective study. SETTING: Surgical ICU. PATIENTS: 35 patients aged 19-79 years, divided into two groups: sepsis (defined by standard criteria) and nonsepsis. MEASUREMENTS AND RESULTS: Studies of proteolytic degradation of H-kininogen showed degradation in both patient groups compared to healthy controls. Functional quantification of prekallikrein showed a reduction of prekallikrein in plasma from both patients groups. Functional quantification of kininogens by a cysteine proteinase inhibitor assay showed no significant differences between the patients and the controls. Immunological levels of H-kininogen and total kininogen were not significantly different from normal plasma. No differences could be detected between the two patient groups in any of the parameters studied. CONCLUSIONS: This study showed that contact activation took place in surgical ICU patients: partial kinin release and consumption of prekallikrein took place in vivo.

High and low molecular weight kininogen and plasma prekallikrein/plasma kallikrein in villous capillaries of human term placenta.

This study examined the expression and presence of components of the kallikrein-kinin system in human term placenta. Immunohistochemical studies localized H-kininogen and plasma prekallikrein/plasma kallikrein to endothelial cells of placental villous capillaries. In larger placental blood vessels and umbilical cord, neither kininogens nor kallikreins were detected. High (H) and low (L) molecular weight kininogen, plasma prekallikrein and plasma kallikrein were detected by Western blot analysis in human term placenta and in maternal and fetal blood, whereas tissue kallikrein was not. Furthermore, mRNA of plasma prekallikrein was detected by reverse transcriptase-polymerase chain reaction (RT-PCR) in placental homogenates, while mRNA of H-kininogen, L-kininogen and tissue kallikrein was not. Because H-kininogen and plasma prekallikrein circulate in a complexed form, we suggest that endothelial cells bind kininogen and plasma prekallikrein in which they are secreted by the fetal liver from fetal blood. The co-localization of kininogen and plasma prekallikrein/plasma kallikrein suggests that kinins could be generated locally in placental capillaries. When released, they may play a role in regulating placental blood flow and transplacental transport of substrates and metabolites.

[Plasma prekallikrein in chronic liver diseases]. / Prekalikreina osoczowa w przewleklych chorobach watroby.

Plasma prekallikrein (PPK) is a single-chain glycoprotein synthesized in the liver. The aim of our study was to evaluate a plasma prekallikrein as the marker reflecting liver protein synthesis in patients with chronic liver diseases. PPK levels have been measured by own modification of amidolytic micro-assay in 43 patients with chronic liver diseases and 37 healthy volunteers as control group. As compared to control group, PPK level was significantly decreased in patients with chronic active hepatitis and with decompensated liver cirrhosis and significantly increased in patients with liver cirrhosis complicated by hepatocellular carcinoma. There was no difference in plasma prekallikrein between patients with compensated liver cirrhosis and controls. The results suggest that PPK might be a useful index for the assessment of residual functional liver mass in patients with chronic liver diseases.

Patients undergoing reconstructive surgery versus unburned children as "controls" in studies of pediatric patients with burns.

A valid control group is an essential part of any patient study. We asked whether burned children returning for reconstructive surgery could be used as "controls" in a study of seven proteolytic elements in the circulation. Functional levels of elastase, plasminogen, prekallikrein, antithrombin, alpha 2-antiplasmin, alpha 2-macroglobulin, and total proteolytic activity in 30 healthy unburned children were compared with levels in 29 patients admitted for reconstructive surgery an average of 7.9 years after burn. The two groups were not statistically different in distributions of sex and race but differed in mean age. Levels of six of the seven parameters were equal in the two groups. However, even when correction for the age difference was done between the groups, alpha 2-macroglobulin in the patients undergoing reconstructive surgery still was significantly less (p < 0.021) than in the unburned group. Therefore caution may be needed when readily available patients undergoing reconstructive surgery are used as "controls" in studies of patients with acute burns.

Prognostic value of clotting and fibrinolytic systems in a follow-up of 165 liver cirrhotic patients. CALC Group.

One hundred sixty-five patients with cirrhosis diagnosed by needle liver biopsy were followed for 2 years to evaluate the relation between clotting factors and survival. Patients with spontaneous bacterial peritonitis, hepatic carcinoma, and cholestatic liver diseases were excluded. Patients were classified as A (n = 34), B (n = 75), or C (n = 56) according to Child-Pugh criteria. During the follow-up 45 patients died of liver failure or gastrointestinal hemorrhage. Nonsurvivor patients had significantly higher values of bilirubin and D-dimer, a marker of fibrinolysis in vivo, lower values of albumin, prothrombin activity, fibrinogen, prekallikrein, factor VII, and a more prolonged activated partial thromboplastin time than survivors. All these variables and Child-Pugh classification were significantly associated with survival in a univariate analysis. Multivariate analysis (Cox's model) showed that only prekallikrein and factor VII were independently predictors of survival. Ninety-three percent of patients with prekallikrein values < 32% died within 32 months of follow-up, whereas factor VII < 34% identified 93% of patients who died within 10 months of follow-up. This study suggests that factor VII is an early predictor of survival and may be a useful test to better identify cirrhotic patients who should be candidates for liver transplantation.