Several components of plasma hemostasis in patients with suppurative processes in the abdominal cavity]

Blood and urine concentrations of antithrombin-III, plasminogen, alpha 2-macroglobulin, alpha 1-antitrypsin, degradation products of fibrinogen-fibrin were studied in patients with abdominal suppuration. Noticeable deviations from the normal values especially marked in the severe process indicated the development of DIC syndrome and renal failure. Heavy combined treatment promoted normalization of the hemostasis shifts and eliminated pyo-inflammatory processes in the abdominal cavity.     

alpha 2-Macroglobulin in patients with obstructive lung disease, with and without alpha 1-antitrypsin deficiency

Serum alpha 2-macroglobulin concentrations were measured in 178 patients with emphysema and 115 control subjects of similar age and sex distribution. The study group included 59 PI type Z patients with alpha 1-antitrypsin deficiency, five with the rare alpha 1-antitrypsin null genotype (PI Q0 or --), and seven with alpha 1-antitrypsin deficiency of the rare PI types MmaltonZ or MduarteZ. Individuals with all types of alpha 1-antitrypsin deficiency were found to have significantly increased serum concentrations of alpha 2M (p less than 0.001). These increased concentrations were associated with all types of alpha 1-antitrypsin deficiency, not only with the PI type Z. The highest alpha 2-macroglobulin concentrations were found in the PI Q0 patients (5 with emphysema, 2 with no lung disease), and these patients had almost no circulating alpha 1-antitrypsin. Raised concentrations of serum alpha 2-macroglobulin were not due to emphysema: 86 patients with emphysema, of PI type M, and the normal control subjects had similar average concentrations of alpha 2-macroglobulin. One control subject with an average alpha 2-macroglobulin concentration of only 41% of normal was found.     

Aberrant forms of alpha(2)-macroglobulin purified from patients with multiple sclerosis

The biochemical properties of alpha(2)-macroglobulin were investigated in four patients with multiple sclerosis and compared to alpha(2)-macroglobulin from healthy controls. An impaired stability of alpha(2)-macroglobulin from the multiple sclerosis patients was demonstrated as a spontaneous conversion to an electrophoretic"fast" form of alpha(2)-macroglobulin upon purification and storage, with a concomitant decrease in functional capacity to inhibit proteinases. The ability to form complexes with proteinases was significantly reduced in alpha(2)-macroglobulin purified from the multiple sclerosis patients. The aberrant molecular arrangements of the protein were not due to proteinase cleavages in the bait regions of alpha(2)-macroglobulin, as demonstrated by gel electrophoresis and protein sequencing. The number of functional thiol esters, however, was reduced in alpha(2)-macroglobulin purified from the multiple sclerosis patients, an observation compatible with the impaired proteinase binding property. Furthermore, differences in isoelectric points were observed between alpha(2)-macroglobulin from the multiple sclerosis patients and alpha(2)-macroglobulin from healthy controls. The results suggest that aberrant forms of alpha(2)-macroglobulin may be present in patients with multiple sclerosis.     

Cationic proteins of human granulocytes. V. Interaction with plasma protease inhibitors.

Several cationic proteins of human granulocytes possess chymotrypsin-like and bactericidal activities. The heat-labile chymotrypsin-like activity is inhibited by serum, owing to complex formation with alpha2-macroglobulin and alpha1-antitrypsin. The molar affinity of the cationic proteins for alpha2-macroglobulin is much higher than that for alpha1-antitrypsin. The results indicate that the molar combining ratios are 1:1 for cationic protein to alpha1-antitrypsin and 2:1 for cationic protein to alpha2-macroglobulin. The proteolytic activity against fibrinogen and casein is inhibited by both alpha2-macroglobulin and alpha1-antitrypsin, whereas the activity against small molecular synthetic substrates is inhibited by alpha1-antitrypsin but not alpha2-macroglobulin. The heat-stable bactericidal action of the cationic proteins against Staphylococcus was also inhibited by serum, probably owing to complex formation with alpha2-macroglobulin and alpha1-antitrypsin.     

Amidolytic and immuno-nephelometric determination of alpha 1-proteinase inhibitor and alpha 2-macroglobulin in serum with calculation of specific inhibitor activities in health and disease

In sera of healthy persons (n = 50) and patients with a variety of diseases (n = 197) the two major proteinase inhibitors, alpha 1-proteinase inhibitor (alpha 1-antitrypsin) and alpha 2-macroglobulin, were measured by two methods: a chromogenic (amidolytic) substrate assay to assess the functional activities, and a laser nephelometric method to determine the immunoreactive concentrations of the respective proteins. The specific proteinase inhibitor activities defined as the number of inhibitor units per g inhibitor protein were calculated. The precision and accuracy of both assays were found to be similar, showing a satisfactory correlation of results for the sera of healthy persons (r = 0.916 for alpha 2-macroglobulin and 0.988 for alpha 1-proteinase inhibitor). In diseased individuals the correlation was lower than in normal persons (0.862 for alpha 2-macroglobulin and 0.907 for alpha 1-proteinase inhibitor). A poor correlation was obtained in patients with liver diseases (r = 0.586 for alpha 1-proteinase inhibitor and 0.852 for alpha 2-macroglobulin). Reference ranges were established for functional and immunological concentrations and for specific inhibitor activities, respectively. Normal values followed a Gaussian distribution. In patients with various diseases including those with acute phase response, the specific inhibitor activities of alpha 1-proteinase inhibitor are reduced significantly; this is because inhibitor activity shows a smaller relative increase than immunoreactivity. Among the various diseases, no significant differences were noted. The specific inhibitor activity of alpha 2-macroglobulin changed significantly only in patients with carcinoma, liver diseases and trauma. Follow up of some patients shows also intraindividual variation of specific proteinase inhibitor activities.     

Concentrations of protease and anti-protease in serum of patients with pancreatic cancer

We measured the concentrations of trypsin, elastase, and three anti-proteases-alpha 1-macroglobulin, alpha 1-antitrypsin, and alpha 1-antichymotrypsin-in serum from 10 patients with pancreatic carcinoma. All 10 showed increased elastase and decreased alpha 2-macroglobulin concentrations, nine had increased alpha 1-antichymotrypsin, and eight had increases in alpha 1-antitrypsin and trypsin. Serial studies during chemotherapy of one patient showed that the protease concentrations decreased during treatment but the concentrations of the anti-proteases remained abnormal.     

Howie's antithrombin activity and thrombin inhibitors in the nephrotic syndrome

A prospective study was carried out on a group of 28 patients affected by nephrotic syndrome in order to compare the antithrombin activity, measured by the technique of Howie, the antithrombin III, measured with chromogenic substrate and by radial immunodiffusion, the alpha 2-macroglobulin and the alpha 1-antitrypsin. An increased level of alpha 2-macroglobulin and of the antithrombin activity, measured by the technique of Howie and a reduction of the level of antithrombin III and of alpha 1-antitrypsin was observed. It is suggested that the increased antithrombin activity is related to the increase of alpha 1-antitrypsin was observed. It is suggested that the increased antithrombin activity is related to the increase of alpha 2-macroglobulin concentration in spite of the simultaneous decrease of antithrombin III.     

Human alpha2-macroglobulin and its antitrypsic and antithrombin activities in serum and plasma.

alpha2-Macroglobulin level, trypsin protein esterase and progressive antithrombin activities were measured in normal and nephrotic sera and plasma. Trypsin protein esterase activity was proportional to the alpha2-macroglobulin concentration in serum and plasma from both normal and nephrotic patients. The results were different, however, with progressive antithrombin activity: in normal plasma, antithrombin III is the main thrombin inhibitor, then alpha2-macroglobulin and alpha1-antitrypsin, whereas in nephrotic syndrome patients, alpha2-macroglobulin is the main thrombin inhibitor.     

Characteristics of certain components of the systemic inflammatory reaction in patients with diffusive peritonitis

The clinical, immunobiochemical and hemostasiological parameters of the systemic inflammatory response (SIR) were studied in 51 patients with diffusive peritonitis. The study showed that lactoferrin, plasminogen/plasmin and alpha 2-macroglobulin can be used, in a set, as informative indices of SIR. A reduced content of lactoferrin, a smaller concentration of proteases inhibitor alpha 2-macroglobulin and a higher relation between plasminogen and plasmin correlate with an unfavorable outcome of the disease beginning from the 1st postoperative day. An activity of Willebrandt's factor is a diagnostic and prognostic marker, which determines a lesion to the endothelium system.     

Course and blood coagulation findings following systemic short-term fibrinolysis in acute myocardial infarct

A series of 16 consecutive patients with acute myocardial infarction was investigated with respect to changes in coagulatory parameters after intravenous short-term treatment with 1,500 000 IU streptokinase (SK) over a period of 90 minutes. Samples for coagulation assays (fibrinogen, thrombin, time activated partial thromboplastin time (aPTT), normotest, thrombin-coagulase time, platelets, antithrombin III, plasminogen and antiplasmin activity, alpha 2-macroglobulin, alpha 1-antitrypsin, factor X a were collected before and immediately after iv SK, and after 4, 8, 12, 24, 36, 48 and 72 hours. Platelets, antithrombin III, factor X a, alpha 1-antitrypsin and alpha 2-macroglobulin showed no changes over the observed period. The concentrations of fibrinogen and the activities of plasminogen and antiplasmin decreased clearly during the first 24 hours, reaching a minimum immediately after SK administration. Thrombin time and aPTT were prolonged for 36 hours, with a maximum in the first hours after lysis. Conclusions: Invasive diagnostic and/or therapeutic procedures during the first 24 hours after SK lysis should be carried out only for a definite, strict indication and under repeated control of the coagulatory status. After 24-36 hours there is a trend to normalisation of haemostasis. After 36 hours, surgery may be performed without fear of complications due to abnormal coagulability.     

Variability of plasma proteins according to molecular size. Long-term and short-term intra-individual variation

The intra-individual variations in serum concentrations of alpha 1-antitrypsin, albumin and alpha 2-macroglobulin were determined using high precision analytical methods. The long-term (3 months) variations were 8.2% for alpha 1-antitrypsin and 2.9% for alpha 2-macroglobulin in five males and five females. The coefficients of variation for albumin were 1.5 and 3.4% for males and females, respectively. In males the long-term variations of albumin and alpha 2-macroglobulin were highly correlated. The short-term (2 days) intra-individual variations in six males were 2.5, 3.8 and 3.4% for alpha 1-antitrypsin, albumin and alpha 2-macroglobulin respectively (coefficients of variation). A diurnal variation was found for albumin with maximal concentrations at 18.00 hours. At 6.00 and 10.00 hours the fractional concentrations of alpha 1-antitrypsin and albumin were lower than for alpha 2-macroglobulin. The variations of the three proteins were positively correlated.     

Use of a method of combined determination of the activity of the trypsin-like proteinases, alpha 1-antitrypsin and alpha 2-macroglobulin, in a gastroenterology clinic]

The authors describe a method for combined measurements of the plasma trypsin-like proteinases, alpha 1-antitrypsin and alpha 2-macroglobulin. The values of these parameters' activities in the plasma of normal subjects and patients with peritonitis and liver cirrhosis are presented. The suggested method may be used for identification of the type of disorders in the system of trypsin-like proteinases and endogenous inhibitors in other diseases.     

Effect of vaccine program on IgG antibody titers for measles,rubella, varicella,and mumps in young adults in Japan: Survey between 2018 and 2021

IntroductionImproved routine immunizations in Japan have led to a reduction in vaccine-preventable diseases. Due to changes in the vaccination program, current young adults received their second vaccination for measles and rubella at different times depending on their birth year, and most of them have not been vaccinated against varicella and mumps. This study investigated the effect of vaccine programs on the immunity of people in Japan.MethodsImmunoglobulin G antibody (IgG) titers against four viruses were determined by enzyme immunoassay in 795 students at a medical university. Titers for measles and rubella were compared according to the students’ birth dates (Group 1: April 2, 1990–April 1, 2000; Group 2: April 2, 2000–).ResultsThe titers of students that satisfied the standard IgG values against measles, rubella, varicella, and mumps were 24.3%, 56.9%, 87.4%, and 47.2%, respectively. Measles and rubella titers were lower in group 2 (estimated mean period from last vaccination, 7.0 years) than group 1 (13.5 years) (p = 0.023 measles, p = 0.037 rubella), indicating attenuation of titers over time. Varicella and mumps antibody prevalence indicated that these infections were endemic, whereas rates of negative titers were higher than those for measles and rubella.ConclusionsIgG titers against viruses were affected by vaccination programs. Declining titers after vaccination should be monitored when the diseases are almost eliminated and boosting is absent. Antibody testing is meaningful for recommending vaccinations and for surveillance of waning immunity. Continuous improvements of vaccination program should be considered to prevent and eliminate diseases.     

Serum elastase inhibitors in cardio-vascular diseases.

Serum elastase inhibiting capacity was measured in three groups: 150 control subjects, 38 hospitalized children without cardiovacular diseases and 202 hospitalized patients suffering from cardiovascular diseases. The values obtained were 53% in control adult subjects and 79% (range 45--90%) in the hospitalized patient groups. The highest levels were recorded at the acute phase of myocardial infarction. The levels of alpha 1-antitrypsin (alpha1-AT) and alpha 2-macroglobulin (alpha2-M) were determined by radial immunodiffusion technique for various levels of inhibitory power. No correlation was found between the inhibitory power levels and the alpha1-AT and alpha2-M levels. This study suggests that other proteins may intervene in the inhibition process of elastolysis.     

The behavior of alpha2-plasmin inhibitor in fibrinolytic states.

Human plasma alpha2-plasmin inhibitor in fibrinolytic states was studied using immunochemical methods and radioiodinated plasminogen. The concentration and activity of plasma alpha2-plasmin inhibitor decreased when urokinase was added to plasma in vitro or infused intravenously in man. The decrease was associated with the appearance of plasmin-alpha2-plasmin inhibitor complex which subsequently disappeared from the circulation in a short time. A decrease of other major inhibitors, such as alpha2-macroglobulin and alpha1-antitrypsin, was not observed when the amount of urokinase added or infused was relatively small, and conversion of plasminogen to plasmin was not extensive. The formation of plasmin-alpha2-macroglobulin complex was observed only when plasma plasminogen was activated with a larger amount of urokinase, and after most of the alpha2-plasmin inhibitor was consumed by forming complexes with plasmin. The formation of plasmin-alpha1-antitrypsin complex was not observed even in the highly activated plasma unless exogenous plasmin was added to the plasma. alpha2-Plasmin inhibitor was the only inhibitor of which the concentration in plasma was significantly decreased in patients with disseminated intravascular coagulation and fibrinolysis among the major plasmin inhibitors in plasma. The most reactive inhibitor for regulating plasma fibrinolysis very likely is alpha2-plasmin inhibitor.     

Neutral proteases of human granulocytes. IV. Interaction between human granulocyte collagenase and plasma protease inhibitors.

Purified human granulocyte collagenase was inactivated by serum through the formation of complexes with alpha 1-antitrypsin and alpha 2-macroglobulin. A molar combining ratio of 1:1 was observed for each inhibitor. The affinity of alpha 2-macroglobulin was about 10 times that of alpha 1-antitrypsin for granulocyte collagenase. The molar concentration of alpha 1-antitrypsin in the blood exceeds that of alpha 2-macroglobulin by about 12 times, so that the inhibitors may be equally important for defence against granulocyte collagenase.     

Diagnostic validity of multivariate combinations of biochemical analytes as markers for rejection and infection in the follow-up of patients with heart transplants

The diagnostic validity of multivariate combinations of alpha 1-antitrypsin, alpha 2-macroglobulin, C-reactive protein, complement C3, complement C4, neopterin in serum, and neopterin in urine as markers for acute cardiac allograft rejection and for differential diagnosis of rejection and infections was investigated in the follow-up of 37 patients with heart transplants. Rejection was diagnosed by endomyocardial biopsy. Infections were classified as 'no infection', 'viral infection', and 'bacterial, fungal or mixed infections'. Although there are significant differences between the mean levels of analytes, multivariate discriminant analysis does not provide an adequate discrimination of rejection and infection states. In separate rejection diagnosis, multivariate combinations of analytes cannot replace endomyocardial biopsy. However, a multivariate combination of alpha 1-antitrypsin, alpha 2-macroglobulin, C-reactive protein, C3, C4 in serum, and neopterin in urine can be used as a screening procedure to reduce the number of endomyocardial biopsies.     

Detection of serum antibody against arrestin from patients with acute disseminated encephalomyelitis.

In our previous study, we found the presence of serum autoantibody against arrestin in patients with multiple sclerosis (MS), while such serum autoantibody was not detected from patients with other neurological diseases and control subjects. We suggested that serum arrestin antibody titers may be useful for the diagnosis and evaluation of the disease's course. In the present study we examined sera from 7 patients, who were initially diagnosed as having acute disseminated encephalomyelitis (ADEM), for the presence of serum antibody against arrestin, in order to study the specificity of the serum antibody among demyelinated diseases. High titers were detected from 2 patients out of 7. One of the patients, a 4 year-old girl, presented with an additional neurological attack during the 6 months after the initial attack, resulting in change of diagnosis to MS. During her disease course the serum titers against arrestin fluctuated in correspondence with the disease's activity. These observations suggest that the presence of serum autoantibody against arrestin may be specific to MS and be helpful for differential diagnosis of ADEM and MS.     

Alpha2-plasmin inhibitor and alpha2-macroglobulin-plasmin complexes in plasma. Quantitation by an enzyme-linked differential antibody immunosorbent assay.

An enzyme-linked differential antibody immunosorbent assay has been developed for the quantification of alpha2-plasmin inhibitor-plasmin and alpha2-macroglobulin-plasmin complexes. In this method the inhibitor-plasmin complex is bound to a surface by an inhibitor-specific antibody, and the plasmin bound to the inhibitor is quantified by a second antibody, rabbit antiplasminogen F(ab')2, labeled with alkaline phosphatase. The hydrolysis of p-nitrophenyl phosphate by the alkaline phosphatase is expressed in femtomoles of plasminogen per milliliter, by reference to a standard plasminogen curve. Inhibitor-enzyme complexes were generated in plasma by the addition of plasmin or of urokinase. The concentration of plasmin added was well below the plasma concentration of alpha2-plasmin inhibitor (1 microM) or of alpha2-macroglobulin (3.5 microM), so that neither inhibitor would be fully saturated with enzyme. Under these conditions increasing amounts of plasmin generated an increase in both alpha2-plasmin inhibitor-plasmin and alpha2-macroglobulin-plasmin complexes. Varying amounts of plasmin were incubated with each of the purified inhibitors in the concentration found in plasma, and the complexes. Varying amounts of plasmin were incubated with each of the purified inhibitors in the concentration found in plasma, and the complexes that formed were quantified by immunoassay. These studies made it possible to quantify the distribution of plasmin between the two inhibitors in plasmin or urokinase-treated plasma. In plasmin-treated plasma, 10% or less of the plasmin bound to both inhibitors was in complex with alpha2-macroglobulin. In contrast, between 19 and 51% of the plasmin generated in urokinase-activated plasma was bound to alpha2-macroglobulin. Thus, major changes in the distribution of plasma were observed, according to whether plasmin was added to plasma or whether plasminogen was activated endogenously. The pattern of inhibitor plasmin complexes generated in vivo by the therapeutic infusion of urokinase was similar to that found for urokinase-activated plasma. 23 normal individuals had low levels of alpha2-plasmin inhibitor-plasmin complexes, whereas six patients with laboratory evidence for disseminated intravascular coagulation demonstrated a 16- to 35-fold increase in he concentration of these complexes. These data indicated that a useful new probe for the study of the fibrinolytic enzyme system had been developed.     

Changes in concanavalin A-reactive proteins in neurological disorders.

Changes of glycosylation of cerebrospinal fluid proteins such as alpha2-macroglobulin, and prostaglandin D synthase were studied by lectin blotting, using concanavalinA, in multiple sclerosis (n = 42) and neuropathies (n = 20) in comparison to neurological controls (n = 22). The concanavalinA-reactivity of alpha2-macroglobulin, which was increased in the neuropathies but not in multiple sclerosis compared to controls, correlated with the total concanavalinA-reactivity in controls and neuropathies but not in multiple sclerosis, indicating that the protein could be abnormally glycosylated in the latter disease. Although the concentration and the concanavalinA-reactivity of prostaglandin D synthase were not significantly different in the three groups, the two parameters correlated only in neuropathies but not in controls or multiple sclerosis, probably due to the high heterogeneity of the protein. These changes deserve to be studied in further detail in view of their potential clinical applications.