Early resuscitation with polymerized bovine hemoglobin reverses acidosis, but not peripheral tissue oxygenation, in a severe hamster shock model

Awake hamsters equipped with the dorsal window chamber preparation were subjected to hemorrhage of 50% of the estimated blood volume. Initial resuscitation (25% of estimated blood volume) with polymerized bovine hemoglobin (PBH) or 10% hydroxyethyl starch (HES) occurred in concert with an equivolumetric bleeding to simulate the early, prehospital setting (exchange transfusion). Resuscitation (25% of estimated blood volume) without bleeding was performed with PBH, HES, or autologous red blood cells (HES-RBCs). Peripheral microcirculation, tissue oxygenation, and systemic hemodynamic and blood gas parameters were assessed. After exchange transfusion, base deficit was -8.6 +/- 3.7 mmol/L (PBH) and -5.1 +/- 5.3 mmol/L (HES) (not significant). Functional capillary density was 17% +/- 6% of baseline (PBH) and 31% +/- 11% (HES) (P < 0.05) and arteriolar diameter 73% +/- 3% of baseline (PBH) and 90% + 5% (HES) (P < 0.01). At the end, hemoglobin levels were 3.7 +/- 0.3 g/dL with HES, 8.2 +/- 0.6 g/dL with PBH, and 10.4 +/- 0.8 g/dL with HES-RBCs (P < 0.01 HES vs. PBH and HES-RBCs, P < 0.05 PBH vs. HES-RBCs). Base excess was restored to baseline with PBH and HES-RBCs, but not with HES (P < 0.05). Functional capillary density was 46% +/- 5% of baseline (PBH), 62% + 20% (HES-RBCs), and 36% +/- 19% (HES) (P < 0.01 HES-RBCs vs. HES). Peripheral oxygen delivery and consumption was highest with HES-RBCs, followed by PBH (P < 0.05 HES-RBCs vs. PBH, P < 0.01 HES-RBCs and PBH vs. HES). In conclusion, the PBH led to a correction of base deficit comparable to blood transfusion. However, oxygenation of the peripheral tissue was inferior with PBH. This was attributed to its negative impact on the peripheral microcirculation caused by arteriolar vasoconstriction.

Evidence of native starch degradation with human small intestinal maltase-glucoamylase (recombinant)

Action of human small intestinal brush border carbohydrate digesting enzymes is thought to involve only final hydrolysis reactions of oligosaccharides to monosaccharides. In vitro starch digestibility assays use fungal amyloglucosidase to provide this function. In this study, recombinant N-terminal subunit enzyme of human small intestinal maltase-glucoamylase (rhMGAM-N) was used to explore digestion of native starches from different botanical sources. The susceptibilities to enzyme hydrolysis varied among the starches. The rate and extent of hydrolysis of amylomaize-5 and amylomaize-7 into glucose were greater than for other starches. Such was not observed with fungal amyloglucosidase or pancreatic alpha-amylase. The degradation of native starch granules showed a surface furrowed pattern in random, radial, or tree-like arrangements that differed substantially from the erosion patterns of amyloglucosidase or alpha-amylase. The evidence of raw starch granule degradation with rhMGAM-N indicates that pancreatic alpha-amylase hydrolysis is not a requirement for native starch digestion in the human small intestine.

Ligand binding induces Cbl-dependent EphB1 receptor degradation through the lysosomal pathway

Eph receptor tyrosine kinases play a critical role in embryonic patterning and angiogenesis. In the adult, they are involved in carcinogenesis and pathological neovascularization. However, the mechanisms underlying their role in tumor formation and metastasis remain to be defined. Here, we demonstrated that stimulation of EphB1 with ephrinB1/Fc led to a marked downregulation of EphB1 protein, a process blocked by the lysosomal inhibitor bafilomycin. Following ephrinB1 stimulation, the ubiquitin ligase Cbl was recruited by EphB1 and then phosphorylated. Both Cbl phosphorylation and EphB1 ubiquitination were blocked by the Src inhibitor PP2. Overexpression of wild-type Cbl, but not of 70Z mutant lacking ligase activity, enhanced EphB1 ubiquitination and degradation. This negative regulation required the tyrosine kinase activity of EphB1 as kinase-dead EphB1-K652R was resistant to Cbl. Glutathione S-transferase binding experiments showed that Cbl bound to EphB1 through its tyrosine kinase-binding domain. In aggregate, we demonstrated that Cbl induces the ubiquitination and lysosomal degradation of activated EphB1, a process requiring EphB1 and Src kinase activity. To our knowledge, this is the first study dissecting the molecular mechanisms leading to EphB1 downregulation, thus paving the way to new means of modulating their angiogenic and tumorigenic properties.

Agreement between HbA1c measured by DCA 2000 and by HPLC: effects of fetal hemoglobin concentrations

In subjects with type 1 diabetes, persisting elevations of fetal hemoglobin (HbF) have been demonstrated. This study evaluated whether HbF levels typically seen in type 1 diabetes (up to 3%) interfere with glycohemoglobin determinations using a common immunologic method (DCA 2000).

Secretion and degradation of glucagon by HIT cells

HIT cells have been widely used to study synthesis and secretion of insulin. It has been assumed that this cell line secretes no other islet hormones. To ascertain whether HIT cells synthesize, secrete, and degrade glucagon, we examined cell extracts for this peptide and compared secretion and degradation of glucagon and insulin during stimulation of the cells by arginine. Glucagon levels in acid extracts of HIT cells were found to be 0.72 +/- 0.15 pmol/mg protein. Both glucagon and insulin were maximally stimulated in a glucagon/insulin molar ratio of 0.029 by arginine concentrations of 25-50 nM, and the concentration of arginine that provided half-maximum responses for both hormones was approximately 3 mM. Diminution of arginine-induced glucagon secretion was caused by somatostatin, a physiological inhibitor of pancreatic islet alpha-cell function. HPLC was used to authenticate the glucagon levels stimulated by arginine for 60 min and measured by RIA. Thirty-six percent of immunoreactive glucagon was found in the fractions representing authentic glucagon, whereas the remaining 64% eluted earlier. Experiments examining the fate of radiolabeled glucagon exposed to HIT cells revealed time-dependent degradation of the radioisotope to earlier eluting forms, which accounted for approximately 50% of the radioactivity by 60 min and was complete by 18 h, indicating that the early peak detected by RIA represented a metabolite of glucagon. Radioisotopic insulin was degraded more slowly with an apparent half-life of approximately 36 h. We conclude that HIT cells are not only able to synthesize, secrete, and degrade insulin, but also much smaller amounts of glucagon.

Hemoglobin Vesicles to Treat Critical Ischemia

The initital purpose for developing artificial oxygen carriers was to replace blood transfusions in order to avoid their adverse effects such as immunologic reactions, transmission of infectious diseases, limited availability and restricted storage conditions. With the advent of new generations of artifical oxygen carriers, a shift of paradigm evolved that considers the artificial oxygen carriers as oxygen therapeutics re-distributing oxygen delivery in the favor of tissues in need. This function may find a particular application in tissues rendered hypoxic due to arterial occlusive diseases. This review, based on a large series of intravital microscopy studies in a hamster skin flap model, outlines the optimal design of hemoglobin vesicles?HbVs?given for the above intention. In summary, the HbV should be of a large diameter, and oxygen affinity, colloid osmotic pressure and viscosity of the HbV solution should be high.