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Effect of α-α diaspirin crosslinked hemoglobin (DCLHb™) on the potency of sodium nitroprusside and nitroglycerine to decrease blood pressure in rats:: A dose-response study

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ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY 2000年第5期28卷 385-396页

作者： Erhart, SM Cole, DJ Patel, PM Drummond, JC Burhop, KE Loma Linda Univ Dept Anesthesiol Loma Linda CA 92350 USA Univ Calif San Diego San Diego CA 92103 USA VA Med Ctr San Diego CA USA Baxter Healthcare Corp Round Lake IL 60073 USA

The nitrovasodilators, sodium nitroprusside and nitroglycerine, effect a dose-dependent decrease in mean arterial blood pressure (MABP) by liberating nitric oxide. alpha-alpha diaspirin crosslinked hemoglobin (DCLHb(TM)) is known to bind nitric oxide. We studied the effect of DCLHb on the potency of sodium nitroprusside (n=36) and nitroglycerine (n=36) to decrease MABP in rats which received 1, 10, 100, 1,000, or 10,000 mg/kg of the DCLHb, or normal saline as the Control. Six doses of sodium nitroprusside or nitroglycerine were given to each rat in a systematically varied sequence. For both drugs, in rats given 1, 10, or 100 mg/kg of DCLHb there were no between groups differences in the change in MABP compared to the Control group. For rats that received 1,000 or 10,000 mg/kg of DCLHb, the potency of nitroglycerine and sodium nitroprusside to decrease MABP was less (p<0.05) than the other groups. These data support the hypothesis that small doses of DCLHb effect a minimal change in the potency of nitrovasodilators to reduce blood pressure. However, they suggest that clinically relevant doses of DCLHb attenuate the vasodilatory ability of sodium nitroprusside and nitroglycerine.

关键词：阿司匹林/类似物和衍生物阿司匹林/药理学血压/药物作用血液代用品/药理学剂量效应关系药物药物评价临床前血红蛋白类/药理学硝酸甘油/药理学硝普钠/药理学大鼠 Sprague-Dawley 血管舒张药动物男(雄)性大鼠

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Inhaled nitric oxide attenuates increased pulmonary artery pressure following diaspirin crosslinked hemoglobin (DCLHb) administration

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ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY 1999年第3期27卷 203-213页

作者： Sefton, W Pudimat, P Bina, S Lojeski, E Mongan, P Muldoon, S Uniformed Serv Univ Hlth Sci Dept Anesthesiol Bethesda MD 20814 USA

Intravenous administration of diaspirin crosslinked hemoglobin (DCLHb) can result in elevated pulmonary and systemic arterial pressures in some mammalian species. This study was designed to evaluate the ability of inhaled nitric oxide (INO) to attenuate elevations in pulmonary artery pressure in a closed-chested swine model. Yorkshire pigs received escalating doses of INO followed by escalating doses of DCLHb or a single dose of DCLHb followed by escalating doses of INO. Systemic and pulmonary arterial pressures were monitored continuously. Significant elevations occurred in systemic and pulmonary arterial pressure following a cumulative dose of 0.1 gm/kg DCLHb. A single dose of 0.3 gm/kg also resulted in elevations of pulmonary and systemic arterial pressure. Inhaled nitric oxide partially reversed the changes in pulmonary but not systemic pressure. These results indicate that INO might be a therapeutic option for humans who may experience increased pulmonary artery pressure following administration of DCLHb.

关键词：投药吸入阿司匹林/类似物和衍生物阿司匹林/药理学血气分析血压/药物作用血液代用品/药理学血红蛋白类/药理学高铁血红蛋白血症/血液一氧化氮/药理学肺动脉/药物作用肺动脉/生理学猪血管舒张药/药理学动物女(雌)性男(雄)性

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Surface modification of diaspirin cross-linked hemoglobin (DCLHb) with chondroitin-4-sulfate derivatives. Part 1

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BIOCONJUGATE CHEMISTRY 2000年第5期11卷 705-713页

作者： Hai, TT Pereira, DE Nelson, DJ Catarello, J Srnak, A Baxter Healthcare Corp Corp Res & Tech Serv Round Lake IL 60073 USA

Synthetic methodology was developed for the preparation of chondrotin-4-sulfate reagents that could be specifically attached to the surface of diaspirin cross-linked hemoglobin (DCLHb), a chemically stabilized human hemoglobin. The surface-modified hemoglobin solutions had a significantly higher colloidal osmotic pressures (COP) than DCLHb. The P-50 of the modified DCLHb was dependent upon the reactive end group of the chondrotin-4-sulfate reagents that was used for the protein modification. Modification of DCLHb with the chondroitin-4-sulfate derivatives containing the maleimide end group 23 provided a hemoglobin with a P50 value of 23 mmHg, while the P-50 of hemoglobins prepared from chondroitin-4-sulfate derivatives containing the aldehyde end group 13 and 18 remained unchanged from that of DCLHb.

关键词：阿司匹林/类似物和衍生物阿司匹林/化学阿司匹林/药理学血液代用品/化学血液代用品/药理学碳水化合物序列软骨素硫酸盐类/化学软骨素硫酸盐类/药理学红细胞聚集/药物作用血红蛋白类/化学血红蛋白类/药理学分子序列数据构效关系表面特性人类

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Complement-mediated acute effects of liposome-encapsulated hemoglobin

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ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY 1999年第1期27卷 23-41页

作者： Szebeni, J Alving, CR Walter Reed Army Inst Res Dept Membrane Biochem Washington DC 20307 USA

Recent studies an liposome-encapsulated hemoglobin (LEH) have indicated that this potential blood substitute can activate the complement (C) system of rats, pigs and man. The reaction can involve both the classical and the alternative pathways, and is mediated, in part, by the binding of natural anti-lipid antibodies to the lipid membrane of liposomes. The significance of these discoveries lies in the fact that C activation appears to be the primary cause of the acute physiological, hematological and laboratory changes that have been observed previously in rats and pigs following the administration of LEH or liposomes, which changes include pulmonary vasoconstriction with decreased cardiac output. In light of the proposed use of LEH as an emergency blood substitute, the latter impairment of cardiopulmonary function may warrant particular circumspection as it could aggravate the clinical state of trauma patients who are prone to develop respiratory distress partly as a consequence of C activation by the injury. Our studies on rats and pigs suggest that the above acute side effects of LEH, including the cardiopulmonary distress, can be efficiently inhibited with soluble complement receptor type I, a specific inhibitor of C activation.

关键词：血液代用品/药理学心排血量/药物作用补体激活/药物作用药物载体血红蛋白类/药理学脂质体/药理学受体补体/代谢血管收缩/药物作用动物人类大鼠

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Antimicrobial activity of a bovine hemoglobin fragment in the tick Boophilus microplus

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JOURNAL OF BIOLOGICAL CHEMISTRY 1999年第36期274卷 25330-25334页

作者： Fogaça, AC da Silva, PI Miranda, MTM Bianchi, AG Miranda, A Ribolla, PEM Daffre, S Univ Sao Paulo Inst Ciencias Biomed Dept Parasitol BR-05508900 Sao Paulo Brazil Inst Butantan Lab Artropodes BR-05503900 Sao Paulo Brazil Univ Sao Paulo Inst Quim Dept Bioquim BR-05508900 Sao Paulo Brazil Univ Sao Paulo Dept Biofis BR-04044020 Sao Paulo Brazil

Antifungal and antibacterial activities were detected in the hemolymph and gut contents of the cattle tick, Boophilus microplus. A peptide with antibacterial activity from the tick gut contents was purified to homogeneity by reversed-phase chromatography. The molecular mass of the purified peptide was 3,205.7 Da, measured by matrix-assisted laser desorption/ionization mass spectrometry. The amino acid sequence was obtained by Edman degradation and showed that the peptide was identical to a fragment of the bovine cu-hemoglobin. A synthetic peptide based on the sequence obtained showed characterization data identical to those of the isolated material, confirming its structure. The synthetic peptide was active in micromolar concentrations against Gram-positive bacteria and fungi. These data led us to conclude that the antibacterial activity detected in tick gut contents is the result of enzymatic processing of a host protein, hemoglobin. This activity may be used by ticks as a defense against microorganisms.

关键词：氨基酸序列抗菌药抗感染药/化学抗感染药/分离和提纯抗感染药/药理学真菌/药物作用革兰氏阳性菌/药物作用血红蛋白类/化学血红蛋白类/分离和提纯血红蛋白类/药理学分子序列数据肽碎片/化学肽碎片/分离和提纯肽碎片/药理学蜱/代谢蜱/微生物学动物牛

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Haemoglobin (Erythrogen™)-enhanced post-thaw growth of cryopreserved cells

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ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY 1999年第2期27卷 163-169页

作者： Azhakanandam, K Anthony, P Lowe, KC Davey, MR Power, JB Univ Nottingham Dept Life Sci Nottingham NG7 2RD England

Supplementation of semi-solid R2 culture medium with a commercial bovine haemoglobin (Hb) solution (Erythrogen(TM)) at 1:50-1:500 (v:v), had beneficial effects on the growth, following cryopreservation, of cells of the Indica rice, Oryza sativa cv. Pusa Basmati 1. The mean absorbance, as assessed by triphenyl tetrazolium chloride reduction, of rice cells at 8 d post-thawing, was increased by up to 60% (P < 0.05), compared to cells recovered in the absence of Hb. Erythrogen(TM) (1:50-1:500 v:v) promoted an increase in biomass, of up to 25% over control (P < 0.05), at 24 d post-thawing. Cell suspensions, re-established by transfer to liquid medium of cells initially thawed and cultured with Erythrogen(TM) for 24 d, exhibited increased (up to 2-fold) growth rates over a subsequent 20-d period, compared to cells recovered without Hb.

关键词：细胞培养的低温保存/方法血红蛋白类/药理学水稻/细胞学水稻/生长和发育

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Synthesis and physicochemical characterization of a series of hemoglobin-based oxygen carriers: Objective comparison between cellular and acellular types

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BIOCONJUGATE CHEMISTRY 2000年第1期11卷 56-64页

作者： Sakai, H Yuasa, M Onuma, H Takeoka, S Tsuchida, E Waseda Univ Adv Res Inst Sci & Engn Dept Polymer Chem Tokyo 1698555 Japan

A series of hemoglobin (Hb)-based O-2 carriers, acellular and cellular types, were synthesized and their physicochemical characteristics were compared. The acellular type includes intramolecularly cross-linked Hb (XLHb), polyoxyethylene (POE)-conjugated pyridoxalated Hb (POE-PLP-Hb), hydroxyethylstarch-conjugated Hb (HES-XLHb), and glutaraldehyde-polymerized XLHb (Poly-XLHb). The cellular type is Hb-vesicles (HbV) of which the surface is modified with POE (POE-HbV). Their particle diameters are 7 +/- 2, 22 +/- 2, 47 +/- 17, 68 +/- 24, and 224 +/- 76 nm, respectively, thus all the materials penetrate across membrane filters with 0.4 mu m pore size, though only the POE-HbV cannot penetrate across the filter with 0.2 mu m pore size. These characteristics of permeability are important to consider-an optimal particle size in microcirculation in vivo. POE-PLP-Hb ([Hb] = 5 g/dL) showed viscosity of 6.1 cP at 332 s(-1) and colloid osmotic pressure (COP) of 70.2 Torr, which are beyond the physiological conditions (human blood, viscosity = 3-4 cP, COP = ca. 25 Torr). XLHb and Poly-XLHb showed viscosities of 1.0 and 1.5 cp, respectively, which are significantly lower than that of blood. COP of POE-HbV is regulated to 20 Torr in 5% human serum albumin (HSA). HES-XLHb and POE-HbV/HSA showed comparable viscosity with human blood. Microscopic observation of human red blood cells (RBC) after mixing blood with POE-PLP-Hb or HES-XLHb disclosed aggregates of RBC, a kind of sludge, indicating a strong interaction with RBC, which is anticipated to modify peripheral blood flow in vivo. On the other hand, XLHb and POE-HbV showed no rouleaux or aggregates of RBC. The acellular Hbs (P-50 = 14-32 Torr) have their specific O-2 affinities determined by their structures, while that;of the cellular POE-HbV is regulated by coencapsulating an appropriate amount of an allosteric effector (e.g., P-50 = 18, 32 Torr). These differences in physicochemical characteristics between the acellular and cellu

关键词：血液代用品/化学合成血液代用品/化学血液代用品/代谢血液代用品/药理学胶剂/化学红细胞聚集/药物作用红细胞/药物作用戊二醛/血液戊二醛/化学血红蛋白类/化学合成血红蛋白类/化学血红蛋白类/代谢血红蛋白类/药理学羟乙基淀粉/化学合成羟乙基淀粉/化学羟乙基淀粉/代谢分子量渗透压氧/投药和剂量氧/血液氧/化学粒子大小聚乙烯二醇类/化学合成聚乙烯二醇类/化学聚乙烯二醇类/代谢黏度人类

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Physiologic responses of cross-linked hemoglobin in endotoxin-treated rats

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ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY 1999年第1期27卷 11-21页

作者： Goff, JM Jackson, MR Taher, M Cutting, MA Alving, BM Krishnamurti, C Walter Reed Army Inst Res Dept Hematol & Vasc Biol Washington DC 20307 USA Walter Reed Army Med Ctr Dept Surg Washington DC 20307 USA

Purified human cross-linked hemoglobin (alpha alpha Hb) as well as recombinant human hemoglobin is undergoing clinical trials in the setting of acute brood loss and perioperative hemodilution. We have previously demonstrated that in rabbits with circulating plasma Hb, such as alpha alpha Hb, infusion of endotoxin (LPS) impairs myocardial contractility which results in hypotension, tissue hypoperfusion and increased mortality. The untoward cardiovascular effects occurring after the combined infusion of LPS and alpha alpha Hb in this model are similar to those reported for other agents that inhibit nitric oxide (NO) availability. To determine if the deleterious effects of alpha alpha Hb and LPS were species specific, we performed similar studies in rats. Anesthetized Sprague-Dawley rats received LPS (4 mg/kg or 40 mg/kg) alone or in combination with alpha alpha Hb (0.7 g/kg). Mean arterial blood pressures (MAP) increased in the group that received alpha alpha Hb alone (105+/-8 to 120+/-7 mm Hg, p=0.2) and a decrease was noted in the groups that received low dose LPS (4 mg/kg, p=0.5) and high dose LPS (40 mg/kg, p=0.016). MAP in rats treated with the LPS at either dose combined with alpha alpha Hb remained unchanged. Levels of urine nitrite, which was measured as a surrogate marker for plasma NO, were significantly decreased at 2 hr in groups that received the combination of alpha alpha Hb and LPS at 4 mg/kg (p=0.022) and 40 mg/kg (p=0.003). No significant decrease was observed in animals treated only with alpha alpha Hb (p=0.21) or LPS (4 mg/kg;p=0.78 and 40 mg/kg;p=0.65). Survival was evaluated during 72 hr in animals that were infused with high dose LPS (40 mg/kg) alone or in combination with alpha alpha Hb and then allowed to recover. The survival of rats treated with LPS alone or the combination was 29% at the end of 24 hr and was 100% for rats receiving only alpha alpha Hb. The data suggest that the toxicity of alpha alpha Hb appears to be a species specific phen

关键词：麻醉血压/药物作用药物协同作用内皮缩血管肽1/血液内毒素类/毒性血红蛋白类/代谢血红蛋白类/药理学低血压/化学诱导低血压/死亡率脂多糖类/药理学一氧化氮/血液亚硝酸盐类/尿大鼠 Sprague-Dawley 动物人类男(雄)性兔大鼠

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The effects of stroma-free and dextran-conjugated hemoglobin on hemodynamics and carotid blood flow in hemorrhaged guinea pigs

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ARTIFICIAL CELLS BLOOD SUBSTITUTES AND IMMOBILIZATION BIOTECHNOLOGY 1999年第1期27卷 49-64页

作者： Caron, A Menu, P Faivre-Fiorina, B Labrude, P Vigneron, C Univ Henri Poincare Fac Pharm Lab Hematol & Physiol F-54001 Nancy France

Hemoglobin solutions are potential resuscitative fluids with volume expanding and oxygen delivery abilities developed to reduce the use of blood transfusion. Most hemoglobin solutions in clinical trials increase transiently arterial pressure by inhibiting nitric oxide-dependent vasodilation. Our objective was to compare the effects on central hemodynamics and carotid blood flow of two hemoglobin solutions after resuscitation from hemorrhage in anesthetized guinea pigs. After anesthesia and instrumentation, severe hemorrhage was induced by withdrawing 50% of the blood volume. Resuscitation was performed after 15 min of hypovolemia with 5% albumin,stroma-free hemoglobin, or hemoglobin conjugated to dextran-benzenetetracarboxylate (Dex-BTC-Hb). The mean arterial pressure (MAP), carotid blood flow (CBF), vascular resistance index and heart rate (HR) were monitored for 3 hours after resuscitation. After hemorrhage, MAP and CBF dropped to 57.6 +/- 4.4% and 58.9 +/- 3.7% of control values respectively. Albumin failed to maintain hemodynamics in the decompensatory phase of shock. Both hemoglobin solutions gave rise to a transient increase in MAP (35%);stroma-free hemoglobin increased the CBF (150%) and resistance index (24%) whereas Dex-BTC-Hb had no effect on CBF and vascular resistances. None of the solutions affected the HR. Modified hemoglobin has attenuated effects on CBF and resistance index compared to stroma-free hemoglobin. This may be due to a balance between the stimulation of nitric oxide synthesis by shear-stress and the inhibition of vasodilation by nitric oxide trapping.

关键词：血压/药物作用颈动脉体/血液供给颈动脉体/药物作用右旋糖酐类/药理学心率/药物作用血红蛋白类/药理学出血/病理生理学局部血流/药物作用间质细胞/生理学动物豚鼠男(雄)性

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O-raffinose cross-linking markedly reduces systemic and renal vasoconstrictor effects of unmodified human hemoglobin

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JOURNAL OF PHARMACOLOGY AND EXPERIMENTAL THERAPEUTICS 1999年第3期288卷 1278-1287页

作者： Lieberthal, W Fuhro, R Freedman, JE Toolan, G Loscalzo, J Valeri, CR Boston Univ Med Ctr Evans Dept Med Renal DivRenal Sect Boston MA 02118 USA Boston Univ Med Ctr Evans Dept Med Div Cardiol Boston MA 02118 USA Boston Univ Med Ctr Naval Blood Res Lab Boston MA USA

The hemodynamic effects of a 20% exchange-transfusion with different solutions of highly purified human hemoglobin A-zero (A(0)) were evaluated. We compared unmodified hemoglobin with hemoglobin cross-linked with O-raffinose. Unmodified hemoglobin increased systemic vascular resistance and mean arterial pressure more than the O-raffinose cross-linked hemoglobin solution (by similar to 45% and similar to 14%, respectively). Unmodified hemoglobin markedly reduced cardiac output (CO) by similar to 21%, whereas CO was unaffected by the O-raffinose crosslinked hemoglobin solution. Unmodified and O-raffinose crosslinked hemoglobin solutions increased mean arterial pressure to comparable extents (similar to 14% and similar to 9%, respectively). Unmodified hemoglobin increased renal vascular resistance 2-fold and reduced the glomerular filtration rate by 58%. In marked contrast, the O-raffinose cross-linked hemoglobin had no deleterious effect on the glomerular filtration rate, renal blood flow, or renal Vascular resistance. The extents to which unmodified and O-raffinose cross-linked hemoglobin solutions inactivated nitric oxide also were compared using three separate in vitro assays: platelet nitric oxide release, nitric oxide-stimulated platelet cGMP production, and endothelium-derived relaxing factor-mediated inhibition of platelet aggregation. Unmodified hemoglobin inactivated or oxidized nitric oxide to a greater extent than the O-raffinose cross-linked hemoglobin solutions in all three assays. In summary, O-raffinose cross-linking substantially reduced the systemic vasoconstriction and the decrease in CO induced by unmodified hemoglobin and eliminated the deleterious effects of unmodified hemoglobin on renal hemodynamics and function. We hypothesize that O-raffinose cross-linking reduces the degree of oxidation of nitric oxide and that this contributes to the reduced vasoactivity of this modified hemoglobin.

关键词：电解质/尿交换输血全血血细胞比容血红蛋白类/分析血红蛋白类/药理学肾/血液供给肾/药物作用一氧化氮/代谢棉子糖/类似物和衍生物棉子糖/分析棉子糖/药理学大鼠 Sprague-Dawley 血管收缩/药物作用动物人类男(雄)性大鼠

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