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Biomedical subjects

A Mackiewicz

Publications and source records attributed to A Mackiewicz.

At least 91 records · Page 5Linked to original sources
PubMed1988

Regulation of rabbit acute phase protein biosynthesis by monokines.

We defined the acute phase behaviour of a number of rabbit plasma proteins in studies (in vivo) and studied the effects of monokine preparations on their synthesis by rabbit primary hepatocyte cultures. Following turpentine injection, increased serum levels of C-reactive protein, serum amyloid A protein, haptoglobin, ceruloplasmin, and decreased concentrations of albumin were observed. In contrast to what is observed in man, concentrations of alpha 2-macroglobulin and transferrin were increased. Co-culture of primary hepatocyte cultures with lipopolysaccharide-activated human peripheral blood monocytes or incubation with conditioned medium prepared from lipopolysaccharide-activated human or rabbit monocytes resulted in dose-dependent induction of serum amyloid A, haptoglobin, ceruloplasmin and transferrin and depression of albumin synthesis, while C-reactive protein synthesis and mRNA levels remained unchanged. A variety of interleukin-1 preparations induced dose-dependent increases in the synthesis and secretion of serum amyloid A, haptoglobin, ceruloplasmin and transferrin and decreased albumin synthesis. Human recombinant tumour necrosis factor (cachectin) induced a dose-dependent increase in synthesis of haptoglobin and ceruloplasmin. In general, human interleukin-1 was more potent than mouse interleukin-1 and tumour necrosis factor. None of the monokines we studied had an effect on C-reactive protein synthesis or mRNA levels. These data confirm that C-reactive protein, serum amyloid A, haptoglobin and ceruloplasmin display acute phase behaviour in the rabbit, and demonstrate that, in contrast to their behaviour in man, alpha 2M and transferrin are positive acute phase proteins in this species. While both interleukin-1 and tumour necrosis factor regulate biosynthesis of a number of these acute phase proteins in rabbit primary hepatocyte cultures, neither of these monokines induced C-reactive protein synthesis. Comparison of these findings with those in human hepatoma cell lines, in which interleukin-1 does not induce serum amyloid A synthesis, suggests that the effect of interleukin-1 on serum amyloid A synthesis may be indirect.

Acute-Phase Proteins↗
PubMed1988

Heterogeneous nature of the acute phase response. Differential regulation of human serum amyloid A, C-reactive protein, and other acute phase proteins by cytokines in Hep 3B cells.

Because a number of different cytokines have been reported to regulate the synthesis of human, murine, and rat acute phase proteins (APP), we studied the effect of cytokines on production of several major human APP in a single system, the human hepatoma cell line Hep 3B. Conditioned medium (CM) prepared from human blood monocytes activated with LPS in the presence of dexamethasone led to substantial induction of serum amyloid A (SAA) and C-reactive protein (CRP) synthesis whereas the defined cytokines IL-1 beta, TNF alpha, and medium from a human keratinocyte cell line (COLO-16), containing hepatocyte-stimulating factor activity, failed to induce these two major APP. Induction of SAA and CRP was accompanied by an increase in concentration of their specific mRNA. Size fractionation of CM from activated monocytes by fast protein liquid chromatography indicated that SAA- and CRP-inducing activity eluted as a single peak with a Mr of approximately 18 kDa. alpha 1-Antitrypsin, which also failed to respond to IL-1 beta or TNF alpha, was induced by both CM and medium from COLO-16 cells. The induction of AT by CM was accompanied by an increase in specific mRNA. Induction of ceruloplasmin and alpha 1-antichymotrypsin and decrease in the synthesis of albumin was achieved by both CM and IL-1 beta. Ceruloplasmin and albumin responded in a comparable fashion to both TNF alpha and medium from COLO-16 cells; the response of ACT to these cytokines was not evaluated. These results indicate that human SAA and CRP are induced in Hep 3B cells by products of activated monocytes but not by IL-1 beta, TNF-alpha, or some hepatocyte-stimulating factor preparations and that a group of heterogeneous mechanisms are involved in the induction of the various human APP.

Acute-Phase Proteins↗
PubMed1987

Monokines regulate glycosylation of acute-phase proteins.

The acute-phase response to inflammatory stimuli, characterized by increased synthesis of acute-phase proteins (APP), is often accompanied by changes in the glycosylation patterns of some of these proteins. While expression of APP genes in hepatocytes is regulated by monokines, mechanisms governing changes in glycosylation are not known. Exposure of human hepatoma cell line Hep 3B to conditioned medium from LPS-activated human monocytes and to medium from the keratocarcinoma cell line COLO-16 led to increased synthesis of alpha 1 proteinase-inhibitor and ceruloplasmin and to alterations of their glycosylation patterns similar to those seen in human serum in various inflammatory states. IL-1, tumor necrosis factor, and hepatocyte stimulating factor I increased synthesis of ceruloplasmin without alterations in the pattern of its glycosylation. These findings demonstrate that altered glycosylation seen in plasma in some inflammatory states can be explained by the effects of monokines on glycosylation in hepatocytes and that gene expression and glycosylation of some APP during the acute-phase response may be regulated by different mechanisms.

Acute-Phase Proteins↗
PubMed1987

Microheterogeneity forms of alpha 1-acid glycoprotein as indicators of rheumatoid arthritis activity.

The microheterogeneity of alpha 1-acid glycoprotein (AGP) has been studied in the sera of 48 patients with rheumatoid arthritis and of 12 healthy individuals. For each rheumatoid patient the disease activity has been assessed and each patient has been assigned to one of four activity grades: I, inactive; II, mildly active; III, moderately active; and IV, severe. Concanavalin A-affinity electrophoresis revealed three microheterogeneity variants of AGP: non-reactive with Con A, weakly reactive with Con A and strongly reactive with Con A. The relative amounts of AGP-variants observed in the healthy donors were similar to those observed in the patients with activity grade I, but differed significantly from patients with grades II, III and IV. The differences between the AGP-patterns of patients with activity grades II, III and IV were also statistically significant.

Arthritis, Rheumatoid↗
PubMed1987

Microheterogeneity of alpha 1-acid glycoprotein in the detection of intercurrent infection in systemic lupus erythematosus.

We evaluated the clinical usefulness of determinations of alpha 1-acid glycoprotein microheterogeneity patterns in distinguishing patients who have active systemic lupus erythematosus (SLE) from those who have SLE with intercurrent infection. We used agarose affinity electrophoresis with concanavalin A (Con A) as a ligand. Results were expressed as reactivity coefficients (RC), which are the ratios of variants reactive with Con A to the variants not reactive with Con A. No significant differences were found between the mean RC (+/- SD) in healthy individuals (1.35 +/- 0.26) and that in patients with various degrees of SLE activity. In contrast, a significantly higher mean RC was found in sera from patients with intercurrent infection (2.70 +/- 0.76) compared with each of the other groups studied (P less than 0.001). An RC greater than 2.25 was found in none of 42 sera from patients without infection and in 15 of 18 sera from patients with infection (sensitivity 83%, specificity 100%). C-reactive protein (CRP) levels were also significantly higher in SLE patients with intercurrent infection than in patients with very active disease (P less than 0.05). Levels of CRP greater than 60 mg/liter were found in 3 of 42 SLE patients without infection and in 8 of 18 patients with infection (sensitivity 39%, specificity 93%). The results show that in SLE patients, the finding of a relative increase in Con A-bound serum alpha 1-acid glycoprotein is a more sensitive indicator of intercurrent infection than is the finding of increased levels of CRP.

C-Reactive Protein↗
PubMed1986

Determination of lectin-sugar dissociation constants by agarose affinity electrophoresis.

Agarose crossed affinity electrophoresis (aff-EP) was employed for the determination of lectin-sugar dissociation constants (Ki). In the first dimension of the aff-EP increasing amounts of sugar (alpha-methyl-D-mannoside) were added to a given concentration of lectin (concanavalin A). Then the electrophoresis was run with alpha 1-acid glycoprotein, alpha 1-antitrypsin and alpha-fetoprotein as markers of lectin-sugar interactions. Mathematical equations for determination of the mechanisms and constants of lectin-sugar-glycoprotein interactions were developed. The mean value of the concanavalin A-alpha-methyl-D-mannoside dissociation constant calculated according to the introduced equations was 0.28 mM. In this system it was also possible to determine lectin-glycoprotein dissociation constants (K). The observed influence of the sugar on lectin-glycoprotein binding might be due to hydrophobic interactions since the addition of nonionic detergent caused reversal of this phenomenon.

Carbohydrates↗
PubMed1986

Studies on microheterogeneity of acute-phase proteins in rheumatoid arthritis by using crossed affinoimmuno-electrophoresis with free concanavalin A.

Microheterogeneity of two acute-phase proteins: orosomucoid (alpha1-acid glycoprotein, AGP) and alpha 1-antichymotrypsin (ACHT) were studied in the sera of 48 patients with rheumatoid arthritis and 12 healthy individuals. Each rheumatoid patient was assigned to one of four activity grades. Cross affinoimmunoelectrophoresis (aff-EP) with free Concanavalin A (Con A) revealed three microheterogeneity variants of AGP and four microheterogeneity forms of ACHT. The relative amounts of AGP-variants and ACHT-variants observed in the healthy donors were similar to those observed in the patients with activity grade I, but they changed with the increase in the grade of rheumatoid activity. The differences were most significant for AGP. The comparison of serum C-reactive protein (CRP) levels with AGP-variant-O-non reactive with Con A showed significant correlation in respective rheumatoid activity grades.

Acute-Phase Proteins↗
PubMed1985

Lectin inhibition system for determination of concanavalin A glycoprotein complexes dissociation constants in agarose affinity electrophoresis.

A new system for lectin-glycoprotein complexes dissociation constants (K) determination is presented. The system is based on agarose affinity electrophoresis where equal lectin (Con A) concentrations are inhibited by variable specific sugar (alpha-methyl-mannoside) amounts. Moreover, the system allows lectin-sugar inhibition constants (Ki) studies. For determination of mechanisms as well as K and Ki values calculations mathematical equations are developed. Values of K for two variants of alpha 1 acid-glycoprotein, two variants of alpha 1-antitrypsin, one variant of alphafetoprotein and Ki for Con A-alpha-MM are calculated according to the introduced equations and compared in a computed system. Moreover, the influence of sugar on lectin-glycoproteins interaction is demonstrated and discussed.

Arthritis, Rheumatoid↗
PubMed1985

Comparison of three immunoassays for C-reactive protein determination.

Three widely used immunoassays for C-reactive protein (CRP) determination (RID. RIEP and LN--Behring) were compared. According to the CRP calcium dependent structural changes or ligand binding properties two buffer systems, Ca2+ rich and Ca2+ depleted, were used in all methods. In RID and RIEP mean values of studied samples were higher when Ca2+ depleted buffer was used (r = 0.94 and r = 0.88, respectively). However, in LN mean values of studied samples were higher when Ca2+ rich buffer was employed (r = 0.923). Comparison of RID and RIEP showed better correlation when Ca2+ depleted buffer was used with r = 0.913 and regression equation y = 0.92 chi + 2.76. Comparison of RID with LN gave similar results as in the case of RIEP and LN. In both cases mean values of studied samples were much higher in LN. Best correlation between RID and LN was obtained when RID was run in Ca2+ depleted buffer and LN performed in Ca2+ rich buffer (r = 0.845, y = 1.25 chi + 18.4). When RIEP and LN were compared highest correlation was obtained when Ca2+ rich buffer in both methods had been used (r = 0.84, y = 1.45 chi + 33.7). However, comparison of both methods run in Ca2+ depleted buffer yielded better regression equation: y = 1.05 chi + 26.1 with r = 0.795.

Arthritis, Rheumatoid↗
PubMed1985

The effect of C-reactive protein on the PHA-induced proliferation of human peripheral blood lymphocytes.

The effect of CRP purified from ascites fluid originated from cancer patients on human peripheral blood lymphocytes was investigated. CRP was purified using 4-step procedure including absorption on Sepharose 4B, phosphocholine-Sepharose, DE-52 ion exchange chromatography and gel filtration on Sephacryl S-200. CRP was able either to inhibit or to enhance PHA-induced lymphocyte proliferation in vitro. The effects of added CRP depended on the time of its addition, dose of CRP and lymphocyte donor.

C-Reactive Protein↗
PubMed1981

Microheterogeneity of alpha-fetoprotein in patient serum as demonstrated by lectin affino-electrophoresis.

Microheterogeneity of alpha-fetoprotein (AFP) present in the sera of 76 patients was studied by lectin affino-immunoelectrophoresis. Seventeen patients had benign liver disorders and the remaining 59 patients were treated for primary or secondary liver cancer or yolk sac tumour. By means of Con A, AFP was divided into two variants, while lentil lectin (LCA) made it possible to separate AFP in three variants. In some patients the relative concentrations of Con A and LCA AFP variants were similar; these patients were believed to produce AFP of the same 'profile'. Fourteen AFP profiles were observed by estimation of the area enclosed by precipitates corresponding to respective AFP variants. It was also possible to estimate the AFP profile on the basis of a simple visual analysis of the electrophoretic plates. The obtained results indicate that the AFP profiles of patients with cancer were variable. In spite of variations of the AFP profile in cancer patients, in most cases it was possible to differentiate primary liver cancer from yolk sac tumour and from liver metastases of cancer. In addition, in two-thirds of hepatoma patients the AFP profile was different from the profile observed in patients with benign liver disorders.

Carcinoma, Hepatocellular↗
PubMed1980

The in vitro production of alpha--fetoprotein variants by human fetal organs.

We have demonstrated the immunological identity of alpha-fetoprotein (AFP) produced in vitro by various fetal organs such as the yolk sac, liver, intestine and kidney. AFP produced in vitro by these organs, present in fetal serum and in samples of amniotic fluid was, however, heterogeneous when studied by lectin affinity immunoelectrophoresis. Results obtained with Concanavalin A were comparable to those obtained with Lentil agglutinin, but Concanavalin A separated AFP into two variants only, while Lentil agglutinin recognised three AFP variants. AFP produced by the yolk sac, intestine and kidney was similar in their proportions of variants and differed clearly from liver AFP. The ratios of AFP variants present in fetal serum were dissimilar to those of yolk sac AFP and of liver AFP. Relative amounts of AFP variants in fetal serum were similar in the first and last trimester of gestation. The proportions of AFP variants present in amniotic fluid in the first trimester of gestation were different from fetal serum, but, at the end of pregnancy, they become identical to those observed in fetal serum. Application of AFP variants for prenatal and cancer diagnosis is discussed.

Amino Acids↗
PubMedDate not supplied

Affinity electrophoresis for diagnosis of cancer and inflammatory conditions.

Affinity electrophoresis, with concanavalin A and Lens culinaris agglutinin as ligands, was applied to study the microheterogeneity of serum proteins, with special emphasis on alpha 1-fetoprotein and alpha 1-acid glycoprotein, two proteins of potential clinical value. A total of 602 samples of serum from patients with various neoplastic and inflammatory conditions were evaluated. Affinity electrophoresis provided useful information for differential diagnosis of cancers of various origins including hepatomas, metastatic liver cancers and yolk sac tumors. The method also proved indispensable for the detection of intercurrent infection in patients with rheumatoid arthritis, systemic lupus erythematosus and scaled burns.

Adult↗
PubMedDate not supplied

Microheterogeneity forms of alpha-fetoprotein present in amniotic fluid.

The microheterogeneity forms of human alpha-fetoprotein present in amniotic fluid were studied in affinity immunoelectrophoresis using two lectins: concanavalin A and Lens culinaris agglutinin. Ninety-seven samples of amniotic fluid collected throughout gestation were obtained from pregnancies in which there was no fetal malformation. Two samples of amniotic fluid from pregnancies complicated by anencephaly and one sample obtained from an empty gestational sac were also studied. The changes in relative concentrations of AFP microheterogeneity forms during gestation are described. It is suggested that concanavalin A is suitable for the demonstration of malformations of the central nervous system in the first and second trimester of pregnancy while in the third trimester it would be of no value. The results of this study suggest that Lens culinaris agglutinin might be suitable for this purpose in the third trimester of gestation, but this needs to be confirmed in a larger study. The optimal concentrations of both lectins were investigated. In addition, a simple method of rocket affinity electrophoresis is also described. The possible mechanisms responsible for the changes in relative concentrations of microheterogeneity forms of alpha-fetoprotein during gestation and in pathological conditions are discussed.

Amniotic Fluid↗
PubMedDate not supplied

Butyric acid enhances in vivo expression of hTNF-alpha in transduced melanoma cell line.

Butyric acid (NaBut) and its derivatives are well-known agents eliciting tumor cell differentiation and apoptosis. In experimental models, NaBut is also used to enhance the efficacy of viral vectors. With the use of B78 murine melanoma cells transduced with the retroviral vector containing human tumor necrosis factor alpha (hTNF-alpha) gene, we investigated the ability of NaBut to increase the cytokine expression. We observed an increase in hTNF-alpha expression in vitro after incubation with NaBut. We also describe that the NaBut pro-drug tributyrin is able to increase hTNF-alpha expression in transduced B78 cells in a tumor vaccination model in mice. This observation strongly suggests a novel potential role for NaBut and its derivatives in tumor therapy. It could be used not only as a therapeutic directly acting on tumor cells but, in parallel, as a genetic vaccine "enhancer".

Animals↗