The pathogenetic role of impaired fatty acid trapping by adipocytes in generating the pleiotropic features of hyperapoB.
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Adipocytes↗
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Systemic blood activation with open and closed venous reservoirs.
In 20 patients undergoing coronary artery bypass grafting, we studied prospectively systemic blood activation, blood loss, and the need for donor blood when using an extracorporeal circuit equipped at random with one of two different venous reservoirs. In 10 patients we used an open venous reservoir system (ORS) consisting of a hard shell venous reservoir with an integral cardiotomy filter, and in 10 patients we used a closed reservoir system consisting of a collapsible venous reservoir and separate cardiotomy reservoir. Concentrations of complement 3a, elastase, thromboxane B2, and fibrin degradation products showed a biphasic course, especially in ORS patients. During bypass, we observed a first peak of levels of complement 3a, thromboxane B2, fibrin degradation products, and elastase, which was higher in ORS patients than in patients with the closed system, because their blood continuously contacted the foreign materials of the filter and air in the open reservoir, which was avoided in the closed reservoir. Intensive blood-foreign material contact also caused the highest (p < 0.05) hemolysis in ORS patients. The larger amount of hemolytic products in ORS patients theoretically resulted in a temporary decrease in capacity of their Kupffer cells to clear endotoxin released after aortic declamping. This theory might explain the significantly (p < 0.01) higher second peak of activated products after declamping that was observed in ORS patients. Due to increased blood activation, the largest (p < 0.001) amount of shed blood loss, greatest (p < 0.05) need for colloid-crystalloid infusion, and largest (not significant) need for donor blood were found in ORS patients (0.8 +/- 0.4 versus 0.2 +/- 0.2 units of packed cells).(ABSTRACT TRUNCATED AT 250 WORDS)
Aged↗
The effect of ASP on the adipocyte of the morbidly obese.
The control of triglyceride synthesis within the adipocyte is not fully understood. Insulin is considered to be the most potent stimulant of triglyceride synthesis. In this paper, we report on the effect of a small (14000 Da), basic (pI 9.0) protein isolated from human serum. This protein has been called acylation stimulating protein (ASP). It is a potent stimulant of triglyceride synthesis in adipocytes from both normal weight and morbidly obese subjects. Its stimulatory effect on adipocytes is both rapid, occurring between 15-30 min after the start of incubation, and prolonged, lasting for up to 3 hr. Compared to insulin, it is sixfold more potent in its effect on triglyceride synthesis. As well as acting on isolated cells, ASP also has a fourfold stimulatory effect on triglyceride synthesis in human adipose microsomes at a concentration of 25 micrograms/ml. This study indicates that ASP is a potent stimulant of triglyceride synthesis and therefore may play a role in the pathogenesis of morbid obesity.
Adipose Tissue↗
Interleukin-1 and osteoarthritis.
Il-1, a multifunctional monokine, can stimulate both synoviocytes and articular chondrocytes to release neutral proteases and prostaglandin E2. It is also capable of promoting bone resorption. Therefore, this molecule (or family of molecules) is likely to play an important role in the mechanism of articular cartilage destruction that occurs in degenerative arthropathies. The synovial tissue itself can produce Il-1 (Catabolin) in some conditions, such as a slight traumatism, so that the presence of local inflammation is not necessary for "Il-1-cartilage" interaction to occur. Fundamental macromolecules of cartilage (collagens, proteoglycans) exert a stimulatory effect on Il-1 production, either as such or in the form of immune complexes. Some activated complement fractions (C3a and C5a) may also be actively involved. Studies on the mechanisms which regulate Il-1 synthesis and release, as well as investigations on the response of target cells to Il-1, are presently fascinating goals that could lead to new strategies in therapeutic research.
Animals↗
Three-year treatment of familial heterozygous hypercholesterolemia by extracorporeal low-density lipoprotein immunoadsorption with polyclonal apolipoprotein B antibodies.
Familial hypercholesterolemia is a disorder of lipid metabolism associated with a highly increased risk for cardiovascular disease. Since in such patients even combined drug therapy often fails to decrease low-density lipoprotein (LDL) cholesterol levels sufficiently, extracorporeal LDL elimination has been developed. We treated eight adult patients with LDL immunoadsorption using antibodies against apolipoprotein B without additional lipid-lowering drug therapy for 3 years; this procedure was performed at weekly intervals. By one treatment session, LDL cholesterol and lipoprotein(a) levels were decreased by 55%. Under regular treatment, mean LDL cholesterol levels of 165 mg/dL between two consecutive treatment sessions could be reached, compared with 522 +/- 24 mg/dL before any treatment. As high-density lipoprotein (HDL) cholesterol levels increased under regular treatment, the LDL/HDL cholesterol ratio decreased from 13.4 to 3.4. Positive influences on plasma and whole-blood viscosity as well as on erythrocyte aggregation also seem to be beneficial with regard to retarding atherosclerosis. Very-low-density lipoprotein (VLDL) levels were reduced by approximately 50% after treatment, accompanied by a marked increase of lipoprotein lipase (LPL) and hepatic triglyceride lipase (HTGL) activity. The effects of LDL apheresis on hemostasis, complement activation transport proteins, and hematological parameters were found to be small. In addition, no side effects amounting to any major clinical relevance occurred in any of the patients. After 3 years of LDL apheresis, a decrease in the frequency of anginal chest pain and ST segment depression on exercise testing and a marked reduction of tendon xanthoma size were observed.
Adult↗
Coordinate regulation of triacylglycerol synthesis and glucose transport by acylation-stimulating protein.
Acylation-stimulating protein (ASP) is the most potent recognized stimulant of triacylglycerol synthesis in human adipocytes. However, its mechanism(s) of action have not yet been elucidated in detail. The present study examines the effects of ASP on membrane transport of glucose and fatty acids in cultured human skin fibroblasts. The data demonstrate that ASP stimulated carrier-mediated glucose transport in a time- and concentration-dependent manner, an effect that was greater than and independent of that observed with insulin. ASP increased the Vmax for glucose transport with no change in the transport Km, suggesting that ASP might result in increased sugar transporters in the plasma membrane. This finding was supported by quantitative Western blot analyses using a monoclonal antibody (G3) that demonstrated an increase in the plasma membrane content of the glucose transporter (Glut 1) with a concomitant decrease in the glucose transporter content of an intracellular membrane fraction. By contrast, ASP had no effect on specific membrane transport of fatty acids. Maximal effects of ASP on triacylglycerol synthesis were demonstrated at saturating levels of both glucose and oleate. Comparable stimulation by ASP in the absence of glucose (with or without pyruvate) was also demonstrated, although the absolute rates of triacylglycerol synthesis were substantially lower. Finally, it was shown that ASP increased the apparent Vmax for triglyceride synthesis without changing its Km. Since ASP will act in the absence of extracellular glucose, ASP must have additional actions, independent of its effect on specific membrane transport of glucose by which it accelerates intracellular triglyceride synthesis.(ABSTRACT TRUNCATED AT 250 WORDS)
Biological Transport↗
Relationship between reduction of complement activation by polysaccharide surfaces bearing diethylaminoethyl groups and their degree of substitution.
The relationships between substitution of hydroxyl (OH) by diethylaminoethyl (DEAE) groups on cellulose membranes and the resulting reduction of complement activation are not clear. As a model, Sephadex has been randomly substituted with DEAE groups. Modification of 10% of the glucose units, i.e. about 3% of the groups, had no noticeable effect on the complement activating capacity of the polymer surface, whereas 20% substitution dramatically reduced it. This result could be explained in part by the fact that Sephadex-specific antibodies are not adsorbed by this modified surface. Thus, they cannot enhance complement activation as they do on Sephadex or cellulose. Comparison of our results with those published on Hemophan led us to the conclusion that a similar effect could occur. However, the degree of substitution of the membrane surface is probably greater than that of the membrane core due to the preparation process. Comparison with Sephadex substituted with other groups, or with other polymers bearing a variable density of OH groups, led us to the conclusion that, as hypothesized by Chenoweth (Artif Organs 1984; 8: 281-287), reduction of the availability of OH groups decreases complement activation, but the relationship is probably not linear. Moreover, replacement by other groups results in additional modulations due to (i) interactions between the groups and proteins responsible for formation or decay of the amplifying C3 convertase and/or (ii) decrease in recognition of the modified surface by antibodies which enhance complement activation on polysaccharides.
Adsorption↗
Splendore-Hoeppli phenomenon in the conjunctiva: immunohistochemical analysis.
PURPOSE: To present two cases of conjunctival lesions exhibiting the Splendore-Hoeppli phenomenon, each with different immunohistochemical findings. DESIGN: Interventional case reports. METHODS: Two young males with conjunctival lesions underwent biopsy. Demographic data and timing of biopsy were extracted from the charts. The biopsy specimens were formalin fixed and paraffin embedded for light microscopy. Immunohistochemical staining using the peroxidase method was carried out on each for IgG, IgM, IgA, the C3 component of complement, and eosinophilic major basic protein. MAIN OUTCOME MEASURES: Presence of positive or negative staining for the various antigens. RESULTS: Both biopsy specimens exhibited the Splendore-Hoeppli phenomenon, a morphologically unique process consisting of an amorphous, eosinophilic material surrounded by epithelioid histiocytes, multinucleated giant cells, lymphocytes, and eosinophils. Two staining patterns were seen. One revealed predominately immunoglobulin deposition, whereas the other revealed primarily eosinophilic major basic protein. This is the first instance we are aware of in which eosinophilic major basic protein was the predominate finding in an ocular specimen. CONCLUSION: The composition of Splendore-Hoeppli phenomenon material varies and may be related to various factors, including timing of biopsy and prior treatment.
Adult↗
Randomized comparison between normothermic and hypothermic cardiopulmonary bypass in pediatric open-heart surgery.
BACKGROUND: The purpose of this study is to investigate the effect of cardiopulmonary bypass (CPB) temperature on myocardial reperfusion injury, oxidative stress, and inflammatory response in pediatric open heart surgery. METHODS: Fifty-nine children (median age 78 months; interquartile range, 39-130) undergoing correction of simple congenital heart defects were randomized to receive either hypothermic (28 degrees C) or normothermic (35-37 degrees C) CPB. Troponin I and 8-isoprostane, complement activation C3a, interleukin (IL) -6, -8, and -10, were measured preoperatively, on removal of the aortic cross clamp, 30 minutes, 6, and 24 hours postoperatively. RESULTS: Troponin I and 8-isoprostane were significantly raised, compared to baseline, in both groups, and remained high at 24 hours. Overall, troponin I and 8-isoprostane levels were 37% and 84% higher in the hypothermic than in the normothermic group, respectively (ratio 1.37, 95% CI 1.00 to 1.88, p = 0.053 and 1.84, 95% CI 1.22 to 2.78, p = 0.0045, respectively), and there was no evidence to suggest the treatment effect changed significantly over the time points measured (p = 0.63). Adjusting for aortic cross-clamp time reduced the effect of hypothermia on troponin (p = 0.18) but not on 8-isoprostane levels (p = 0.0028). The C3a, IL-6, and IL-8 release was similar in the two groups. The IL-10 release between the groups changed over time (p = 0.059) and examining differences at individual time points highlighted a statistically significant difference at the end of the cross-clamp time (p = 0.0079). CONCLUSIONS: Normothermic CPB is associated with reduced oxidative stress compared with hypothermic CPB, and similar myocardial reperfusion injury and whole body inflammatory response, in children undergoing open heart surgery. A larger study with clinical outcomes as primary end points is now warranted.
Biomarkers↗
Impact of the off-pump Fontan procedure on complement activation and cytokine generation.
BACKGROUND: To investigate whether the Fontan procedure in an off-pump fashion is less invasive in terms of the systemic inflammatory reaction. METHODS: Plasma levels of complement and cytokines were measured during and after the Fontan procedure in consecutive 38 patients. Of these, 16 underwent the extracardiac method without use of cardiopulmonary bypass (off-pump group), while the machine was used in the remaining 22 because of intracardiac maneuvers concomitantly needed (CPB group). RESULTS: There was no difference, between these two groups, in any value of plasma complement or cytokines after anesthetic induction. Immediately after commencement of the Fontan circulation, however, plasma concentration was significantly lower in the off-pump group for activated complement 3 (C3a), interleukin-6, interleukin-8, and polymorphonuclear elastase. The C3a value was also lower at 2 hours later in the off-pump group than in the CPB group. Furthermore, the values even stayed within the normal ranges, in the off-pump group, for tumor necrosis factor alpha (TNFalpha) and thrombomodulin. A pulmonary venous oxygen tension divided by an inspired oxygen tension (PpvO2/FiO2) ratio immediately after commencement of the Fontan circulation was 528 +/- 93 mm Hg (410 to 580 mm Hg) in the off-pump group, and 258 +/- 167 mm Hg (86 to 540 mm Hg) in the CPB group (p = 0.01). Duration of drainage for fluid sequestration was shorter in the former group (6.3 +/- 0.7 days versus 13.9 +/- 2.5 days, p = 0.02). CONCLUSIONS: Inflammatory reactions were attenuated when the Fontan procedure was employed in an off-pump fashion compared with the usual procedure on bypass.
Adolescent↗
Markers for sepsis diagnosis: what is useful?
Timely diagnosis of the different severities of septic inflammation is potentially lifesaving because therapies that have been shown to lower mortality should be initiated early. Sepsis and severe sepsis are accompanied by clinical and laboratory signs of systemic inflammation but patients with inflammation caused by noninfectious causes may present with similar signs and symptoms. It is important to identify markers for an early diagnosis of sepsis and organ dysfunction. This article presents currently interesting sepsis biomarkers. Other novel markers and their potential role are discussed.
Acute-Phase Proteins↗
Hemodynamic effects of cardiotomy suction blood.
OBJECTIVE: Cardiac surgery induces a systemic inflammatory activation, which in severe cases is associated with peripheral vasodilation and hypotension. Cardiotomy suction blood contains high levels of inflammatory mediators, but the effect of cardiotomy suction blood on the vasculture is unknown. We investigated the effect of cardiotomy suction blood on systemic vascular resistance in vivo and whether cell-saver processing of suction blood affects the vascular response. METHODS: Twenty-five patients undergoing coronary surgery (mean age, 68 +/- 2 years; 80% men) were included in a prospective randomized study. The patients were randomized to retransfusion of cell-saver processed (n = 13) or cell-saver unprocessed (n = 12) suction blood during full cardiopulmonary bypass. Mean arterial blood pressure was continuously registered during retransfusion, and systemic vascular resistance was calculated. Plasma concentrations of tumor necrosis factor alpha, interleukin 6, and complement factor C3a were measured in suction blood. RESULTS: Retransfusion of cardiotomy suction blood induced a transient reduction in systemic vascular resistance in all patients. The peak reduction was significantly less pronounced in the group receiving cell-saver processed blood (-12% +/- 2% vs -28% +/- 3%, P = .001). There was a significant correlation between tumor necrosis factor alpha concentration in retransfused cardiotomy suction blood and peak reduction of systemic vascular resistance (r = 0.60, P = .002). CONCLUSIONS: The results suggest cardiotomy suction blood is vasoactive and might influence vascular resistance and blood pressure during cardiac surgery. The observed vasodilation is proportional to the inflammatory activation of suction blood and can be reduced by processing suction blood with a cell-saving device before retransfusion.
Aged↗
Platelet storage lesion: an update on the impact of various leukoreduction processes on the biological response modifiers.
Currently, platelet concentrates are produced either from pooled buffy which are leukoreduced during processing, by various types of WBC removal filters and several apheresis technologies, which are leukocyte-reduced during collection with or without filtration. It is therefore important to define the impact of various leukocyte-removal processes on the acceleration of platelet storage of lesion and cellular apoptosis/necrosis. This overview briefly highlights the effects of exposure to artificial surfaces, during apheresis leukoreduction processes and platelet storage bags on the development of the platelet storage lesion that may contribute to transfusion reactions. The results obtained from three plateletpheresis technologies are compared with data from a "like with like" study on buffy-coat-derived platelet concentrates, using three types of platelet filter/pack assemblies. Emphasis is placed on the combined preparative methods and storage bags on generation/removal of: kallikrein/kinin; activated complement, leukocytes and platelet-derived cytokines and the development of cellular injuries, measured by the release of Annexin V. There was no systematic evidence of significant cellular fragmentation caused by filtration, but the combined preparative methods and storage bags appears to have some impact on the rate of release of soluble HLA. Moreover, large variability was observed between and within groups, in terms of various laboratory markers of biocompatibility and major biological response modifiers, indicating that much still remains to be done on various aspects of quality improvement to fully abrogate platelet concentrates associated transfusion reactions.
Annexin A5↗
Near completely humanized liver in mice shows human-type metabolic responses to drugs.
Human hepatocytes were transplanted into urokinase-type plasminogen activator-transgenic SCID mice (uPA/SCID mice), which are immunodeficient and undergo liver failure. The transplanted cells were characterized in terms of their in vivo growth potential and functions. The human hepatocytes progressively repopulated the murine host liver. However, the recipients died when the replacement index (RI) of the human hepatocytes exceeded 50%. The hosts (chimeric mice) survived at RI >50% when treated with a drug that has anti-human complement factor activity, and these mice developed livers with RI values as high as 96%. In total, 36 chimeric mice were generated, and the rate of successful engraftment was as high as 92%. The yield of chimeric mice with RI >70% was 32%. The human hepatocytes in the murine host liver expressed mRNAs for a variety of human cytochrome P450 (hCYP) subtypes, in a manner that was similar to the donor liver. The mRNAs for hCYP3A4 and hCYP1A1/2 were induced in the liver in a CYP type-specific manner when the mice were treated with rifampicin and 3-methylcholanthrene, respectively. These results indicate that human hepatocytes that propagate in mice retain their normal pharmacological responses. We conclude that the chimeric mouse developed in the present study is a useful model for assessing the functions and pharmacological responses of human hepatocytes.
Adolescent↗
Clinical performance and biocompatibility of poly(2-methoxyethylacrylate)-coated extracorporeal circuits.
BACKGROUND: Poly(2-methoxyethylacrylate) is an amphiphilic organic polymer consisting of a hydrophobic backbone with pendant hydrophilic groups that has been reported to reduce protein and platelet adsorption in in vitro and ex vivo studies. METHODS: Sixty patients undergoing three-vessel coronary artery bypass grafting were divided into two equal groups. Group 1 had operation with Capiox poly(2-methoxyethylacrylate) coated SX18R oxygenators with noncoated circuits, and group 2 had operation with all noncoated circuits. Hemodynamic variables, blood and urine test results, hematologic variables, complement fractions, C3a and C4d, and interleukin-6 levels were documented preoperatively (T1), on cardiopulmonary bypass (T2), before cessation of cardiopulmonary bypass (T3), after protamine sulfate reversal (T4), and on the first postoperative day (T5). Protein electrophoresis was performed at T1 and T5. Blood cell adhesion and aggregation on fibers were analyzed with optical microscopy, and desorbed protein was evaluated quantitatively by a spectrophotometer using samples obtained when the oxygenators were dismantled after cardiopulmonary bypass. RESULTS: Platelet counts in group 1 demonstrated significant differences at T3, T4, and T5 (p < 0.05) versus group 2 and white blood cell counts in group 1 versus group 2, at counts T4 and T5. Albumin levels were significantly better preserved in group 1 at T4, and T5 and fibrinogen levels, at T3 and T5 (p < 0.05). On electrophoresis, the postoperative albumin level was 57.9% +/- 3% in group 1 versus 50.2% +/- 3% in group 2 (p < 0.05). Postoperative hemorrhage was 452 +/- 35 mL in group 1 and 612 +/- 35 mL in group 2 (p < 0.05). Duration of intubation was significantly lower (p < 0.05) in group 1, as was need of blood transfusion (p < 0.01). More platelet adhesion and aggregation were demonstrated on noncoated oxygenator fibers. The amount of desorbed protein was 0.13 +/- 0.01 mg/dL versus 0.012 +/- 0.001 mg/dL (p < 0.001) on noncoated versus coated fibers, respectively. CONCLUSIONS: Poly(2-methoxyethylacrylate)-coated oxygenators reduce platelet adhesion, platelet aggregation and protein adsorption. This surface provides a better perioperative clinical status through platelet-, albumin-, and fibrinogen-sparing effects.
Acrylates↗
Acylation-stimulating protein (ASP) regulates glucose transport in the rat L6 muscle cell line.
Acylation-stimulating protein (ASP), a human plasma protein, is a potent stimulator of triglyceride synthesis and glucose transport in both human adipocytes and fibroblasts. The purpose of the present in vitro study was to examine the effect of ASP on glucose transport in muscle cells. ASP stimulated 2-deoxy-glucose transport (2-DG) in differentiated rat L6 myotubes in a time (30 min to 24 h) and concentration dependent manner (97% increase). The magnitude of the ASP effect on glucose transport was comparable to the time- and concentration-dependent effects seen with insulin (125% increase), but was additive to insulin, pointing to involvement of differential signalling pathways. ASP stimulation was dependent on cell differentiation in that glucose transport increased by only 12% in myoblasts, comparable to the effect of insulin in myoblasts (15% increase) demonstrating selective responsiveness of the differentiated myotubes to ASP and insulin. The mechanism for the ASP induced increase in glucose transport was also examined. ASP increased the Vmax for 2-DG transport by 183% (4.02 vs. 1.42 nmol/mg cell protein/30 s; ASP vs. Control, respectively). This could be explained by an increased translocation of glucose transporters (GLUT 1, GLUT 4 and GLUT 3) to the plasma membrane surface as demonstrated by Western analysis (+43% P < 0.05, +30% P < 0.05, and +49% P < 0.05, respectively). The effects of ASP were equal to those of insulin (+47%, +26% and +53% for GLUT 1, GLUT 4 and GLUT 3, respectively) and in all cases were paralleled by comparable glucose transport increases under the same incubation conditions. After long-term stimulation (24 h), Western analysis indicated that ASP had a permissive effect on insulin stimulated increases in total GLUT3 and GLUT4 cellular transporter content. These results suggest that muscle is also responsive to ASP and that ASP may play a role in glucose metabolism in both muscle and adipose tissue.
Animals↗
Human acylation stimulating protein enhances triacylglycerol biosynthesis in plant microsomes.
Diacylglycerol acyltransferase has a universal role in catalyzing the acyl-CoA-dependent formation of triacylglycerol in microorganisms, animals and plants. Acylation stimulating protein, from human blood, is known to enhance diacylglycerol acyltransferase activity and triacylglycerol biosynthesis in human adipocytes. In the current study, acylation stimulating protein was also shown to enhance diacylglycerol acyltransferase activity in microsomes from cell suspension cultures of oilseed rape. Enzyme stimulation occurred over the pH range of 6-9 but the degree of stimulation decreased with increasing ionic strength at pH 7.4. Varying acyl-CoA concentration did not affect the degree of stimulation. Membranes from triacylglycerol producing cells in plants and humans may have similar binding sites for acylation stimulating protein which have been preserved during molecular evolution. The results suggest that human acylation stimulating protein may be useful in modifying lipid biosynthesis in plants.
Acyl Coenzyme A↗
ApoE phenotype influences plasma ASP in hyperapoB subjects.
Acylation stimulating protein (ASP) is increased in cardiovascular patients who often present with dyslipidemias. The aim of the present study was to examine the influence of apolipoprotein E (apoE) phenotype and lipids on ASP. Plasma ASP, lipids and apoE phenotype were measured in 407 subjects and separated according to the 75th percentile of apolipoprotein B (apoB) into a HyperapoB (HB) group (apoB=152+/-34 mg/dl, 117 men, 80 women) and a normal apoB (NB) group (apoB=88+/-19 mg/dl, 126 men, 84 women). Triglyceride (TG), cholesterol and LDL cholesterol were significantly increased in HB versus NB but there was no difference in age or body mass index (BMI). HB had increased ASP (42%>75th percentile, median=48.4 nM, P<0.001) versus NB (36.5 nM). There was no difference in ASP in NB with any apoE3 variant (E3/3=41 nM, n=98; E3/4=46 nM, n=55; E3/2=50 nM, n=41), but ASP was increased in E2/2 (126 nM, n=9), and E4/4 (186 nM, n=5, P<0.001 ANOVA). In HB, ASP was increased in three apoE phenotypes: E2/4 (209 nM, n=6), E2/2 (135 nM, n=6) and E4/4 (189 nM, n=26), P<0.001 ANOVA relative to the other apoE phenotypes (E3/3=50 nM, n=102; E3/4=41 nM, n=40; E3/2=87 nM, n=17) with a wide range of values. By stepwise regression analysis, the best model that predicted ASP was: [plasma non-esterified fatty acid (NEFA)+TG+cholesterol], where r=0.407, P=0.001. These data suggest that apoE phenotype may potentially influence ASP, although primarily in rare apoE phenotypes.
Apolipoproteins B↗