Biomedical subjects
D C Jackson
Publications and source records attributed to D C Jackson.
Synthesis of a new template with a built-in adjuvant and its use in constructing peptide vaccine candidates through polyoxime chemistry.
Synthetic lipopeptides are showing promise as vaccine candidates, but until now it has been very difficult to prepare them in homogeneous form. We describe the synthesis and characterization of a new water-soluble, four-branched template with a built-in lipophilic adjuvant (Pam3Cys). Through the use of oxime chemistry, we attached four copies of an unprotected influenza virus peptide and characterized the product (13 kDa) by reversed-phase HPLC and electrospray ionization mass spectrometry. Several other such constructions were made using the new template and different peptides. We seem to have a general method for making synthetic lipopeptides in homogeneous form.
Acid-base state and intermittent breathing in the torpid bat, Eptesicus fuscus.
The effects of intermittent breathing on acid-base state and blood gases were characterized in the torpid bat, Eptesicus fuscus, during steady-state torpor between body temperatures (Tb) of 5 and 37 degrees C. Arterial blood samples were taken from indwelling catheters without disturbing the torpid state. Arterial pH (pHa) of samples taken without knowledge of ventilatory state rose by 0.15 units from 37 to 5 degrees C with a delta pHa/delta Tb slope over this range of -0.0055 U/degrees C. However, at and below Tb = 20 degrees C, Eptesicus fuscus breathes intermittently with typical apneic periods of 40-150 min and 4-12 min at 10 and 20 degrees C, respectively. Samples taken at the end of a ventilatory bout and near the end of an apneic period at Tb = 20 degrees C revealed cyclic changes in pH (from 7.49 +/- 0.02 to 7.34 +/- 0.01), PO2 (from 96.6 +/- 3.4 to 30.8 +/- 3.9 Torr), and PCO2 (28.2 +/- 1.4 to 45.9 +/- 1.5 Torr). Between 10 and 37 degrees C, end-ventilatory pHa varied inversely with temperature with a delta pHa/delta T slope of -0.011 U/degrees C. Because intermittent breathing is common to many animals during hibernation, these results demonstrate the importance of coordinating blood sampling with ventilatory state for a reliable interpretation of acid-base regulation under these conditions.
Ventilatory response to hypoxia and hypercapnia in the torpid bat, Eptesicus fuscus.
Ventilatory pattern and ventilatory responses to hypercapnia and hypoxia were investigated in torpid big brown bats at body temperatures of 5, 10, 20, 30 and 37 degrees C. The pattern of breathing at temperatures below 30 degrees C was intermittent, consisting of rhythmic breathing bouts separated by apneic periods with occasional sporadic, non-rhythmic breathing episodes. Overall ventilation (Ve) was matched consistently to overall oxygen consumption (MO2) over the entire range of temperatures with a mean air convection requirement (Ve/MO2) of 1.28 L/mmol. However, calculating the air convection requirement using only oxygen uptake acquired during ventilation yielded an ectotherm-like temperature relationship. Ventilation was stimulated at all temperatures by either increased inspired CO2 or decreased inspired O2. At 20 degrees C, graded hypercapnic stimulation increased the duration of the rhythmic bouts and decreased the duration of apneas until at high CO2 (greater than 3%) breathing was continuous. Hypoxic stimulation below about 7% O2 increased ventilation by selectively increasing the non-rhythmic ventilations and decreasing rhythmic bouts.
Analysis of the interaction between a synthetic peptide of influenza virus hemagglutinin and monoclonal antibodies using an optical biosensor.
The interaction between two monoclonal antibodies and their corresponding Fab' fragments with a synthetic peptide, corresponding to the C-terminal 23 residues of the HA1 chain of influenza virus hemagglutinin against which they were generated, has been examined using an optical biosensor employing the detection principal of surface plasmon resonance (Pharmacia BIAcore). The data obtained has been analysed in detail by linear transformation of the primary data and nonlinear regression analysis, as well as by analysis of equilibrium binding data. The 2/1 antibodies and their Fab' fragments displayed higher affinity than the corresponding 1/1 proteins. The IgGs were found to have equilibrium association constants (KA) 10-20-fold higher than the corresponding Fab' fragments. This appears largely to be due to differences in the dissociation rate constant (kd) and probably reflects increased avidity due to bivalent binding.
The physiology of hibernation among painted turtles: the Eastern painted turtle Chrysemys picta picta.
Eastern painted turtles (Chrysemys picta picta) from Connecticut were submerged at 3 degrees C in normoxic and anoxic water to simulate potential respiratory environments within their hibernacula. Those in normoxic water could survive submergence for at least 150 d, while those in anoxic water could survive for a maximum of about 125 d. Turtles in normoxic water developed a slight metabolic acidosis as plasma lactate accumulated to about 50 mM in 150 d, while anoxic turtles developed a severe lactic acidosis as plasma lactate reached about 200 mM in 125 d; there was no respiratory acidosis in either group. Plasma [Na+] changed little in either group, [Cl-] fell by about one-third in both, and [K+] increased by about fourfold in anoxic turtles but only slightly in those in normoxic water. Total plasma magnesium and calcium increased profoundly in anoxic turtles but moderately in those in normoxic water. Consideration of charge balance indicates that all major ions were measured in both groups. Plasma glucose remained unchanged in anoxic turtles until after about 75 d of submergence, when it increased and continued to increase with the duration of anoxia, with much variation among individuals; glucose remained unchanged throughout in turtles in normoxic water. Hematocrit doubled in 150 d in turtles in normoxic water; in anoxic turtles, an initial increase was no longer significant by day 100. Plasma osmolality increased markedly in anoxic turtles, largely because of accumulation of lactate, but anoxic turtles only gained about half the mass of turtles in normoxic water, who showed no increase in osmolality. The higher weight gain in the latter group is attributed to selective perfusion and ventilation of extrapulmonary gas exchange surfaces, resulting in a greater osmotic influx of water. The physiologic responses to simulated hibernation of C. picta picta are intermediate between those of Chrysemys picta bellii and Chrysemys picta dorsalis, which correlates with the severity of the winter each subspecies would be expected to encounter.
Lactic acid buffering by bone and shell in anoxic softshell and painted turtles.
We tested two hypotheses: first, that the inferior anoxia tolerance of the softshell turtle, Apalone spinifera, compared to the western painted turtle, Chrysemys picta bellii, is related to its less mineralized shell, and second, that turtle bone, like its shell, stores lactate during prolonged anoxia. Lactate concentrations of blood, hindlimb bone, and shell were measured on normoxic Apalone and Chrysemys and after anoxic submergence at 10 degrees C for 2 and 9 d, respectively. Blood and shell concentrations of Ca(2+), Mg(2+), Na(+), K(+), and inorganic phosphate (P(i); for shell only) were also measured. Because a preliminary study indicated lactate distribution in Chrysemys throughout its skeleton during anoxia at 20 degrees C, we used hindlimb bones as representative skeletal samples. Apalone shell, though a similar percentage of body mass as Chrysemys shell, had higher water content (76.9% vs. 27.9%) and only 20%-25% as much Ca(2+), Mg(2+), CO(2), and P(i). When incubated at constant pH of 6.0 or 6.5, Apalone shell powder released only 25% as much buffer per gram wet weight as Chrysemys shell. In addition, plasma [Ca(2+)] and [Mg(2+)] increased less in Apalone during anoxia at an equivalent plasma lactate concentration. Lactate concentrations increased in the shell and skeletal bone in both species. Despite less mineralization, Apalone shell took up lactate comparably to Chrysemys. In conclusion, a weaker compensatory response to lactic acidosis in Apalone correlates with lower shell mineralization and buffer release and may partially account for the poorer anoxia tolerance of this species.
Transcatheter electrocoagulation: a therapeutic angiographic technique for vessel occlusion.
Previous work has shown that localized electrocoagulation of blood vessels using direct current applied by catheter placed electrodes is feasible. In vitro and in vivo experiments showed that clot size and thrombosis of vessels are directly related to the product of the amount and duration of the current. Surgically created splenic hemorrhage was well controlled in 9 of 10 animals. The vessels remained permanently occluded after electrocoagulation in 8 of 9 animals. Pathological studies of all the vessels showed intimal damage at the sites of occlusion. These experiments indicate that clot formation may be a product of both intimal damage and platelet attraction to the positive electrode. The technique has advantages over embolization since no foreign material is injected and a localized clot is produced at the tip of the guidewire. There are certain disadvantages and further refinements must be developed to establish transcatheter electrocoagulation as a useful clinical technique for vessel occlusion.
Transcatheter electrocoagulation: experimental evaluation of the anode.
Transcatheter electrocoagulation using direct current has been shown to be an effective method for vessel occlusion. To date, the stainless steel anode has not been entirely satisfactory. Thus, an evaluation of bipolar platinum electrodes and a comparison of monopolar platinum and stainless steel electrodes have been performed. Experimental results have shown that both monopolar and bipolar platinum electrodes caused perforation in most cases. At the present time, we have not found a satisfactory replacement for the unipolar stainless steel anode.
The aortogram and gastrointestinal bleeding.
Three cases of gastrointestinal hemorrhage are reported which illustrate that the bleeding site may be missed angiographically if the study is limited to celiac and mesenteric angiograms. An abdominal angiogram must be performed as part of the radiological investigation of gastrointestinal bleeding.
Aortic and iliac graft fistulae.
Sixteen patients with aortic and iliac graft fistulae were diagnosed from a series of over 886 abdominal aortic reconstructive operations. A preoperative diagnosis was established in 12, eleven by radiographic studies. Angiography and barium intestinal studies were the most helpful. The two most important features in diagnosis are a high index of suspicion and an aggressive radiographic approach.
A synthetic peptide of influenza virus hemagglutinin as a model antigen and immunogen.
This paper reviews our studies on a 24-residue synthetic peptide representing part of the amino acid sequence of the influenza virus hemagglutinin. We have used this peptide as a model antigen to define short sequences and individual amino acid residues involved in and critical for interaction with antibody and with T cells. These studies provide insight into the way in which an immunogen is viewed by the immune system and also the minimum requirements necessary for the expression of immunogenic and antigenic activity. This information is helping us in exploiting synthetic peptides in the construction of designer immunogens which have potential as candidate vaccines.
Equilibrium analysis of the interaction between a synthetic peptide of influenza virus hemagglutinin and monoclonal antibodies.
The affinity of interaction between two monoclonal antibodies and a synthetic peptide representing the C-terminal 23 residues of the heavy chain (HA1) of influenza virus hemagglutinin were determined using an air-driven ultracentrifuge. The technique makes use of common laboratory equipment and is based on sound theoretical principles. Because the method does not rely on the solid-phase immobilisation of one of the interacting species, it circumvents problems associated with ELISA-like assays, which, in the case of peptides, may involve the immobilisation of ligand through association of amino acid residues necessary for recognition by antibody. The technique should be applicable to the study of a wide range of ligand-acceptor systems. Because only one of the reagents needs to be pure to allow labelling, prior purification of the biological receptor is not necessary. The method also lends itself to inhibition experiments in which the effects of various homologs on the binding event can be examined in a way which permits an evaluation of potential agonists and antagonists.