Myocardial protection with cold blood potassium cardioplegia during prolonged aortic cross-clamping. Laboratory evaluation.
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Between September 1989 and May 1994, 3 patients with aortic dissection and one with atherosclerotic total aortic aneurysm associated with annuloaortic ectasia underwent successful staged operation for aneurysm of the entire aorta and aortic regurgitation. A composite graft was used for total aortic root replacement. Carbrol and Piehler techniques, Carrel patch and saphenous vein grafting were employed for coronary artery reconstruction. En bloc arch reconstruction was performed in one patient and three vessels graft replacement in 3 patients under hypothermic separate cerebral perfusion. Combined antegrade with retrograde oxygenated crystalloid cardioplegia and terminal warm blood cardioplegia were used for myocardial protection during prolonged aortic cross clamping in a simultaneous total aortic root and arch replacement. Elephant trunk was used at the distal arch anastomosis in 3 patients and useful for following thoracoabdominal surgery. In 3 patients, separate perfusion of upper and lower body technique with moderate hypothermia was employed and seemed to be useful in the patients who require extensive thoracoabdominal replacement to prevent spinal cord injury. All patients had no major complications and have been well.
Heterotrimeric guanine nucleotide-binding proteins are important signaling molecules composed of an alpha, beta, and gamma subunit. The beta subunits must form dimers with gamma subunits to function. Several subtypes of beta and gamma have been identified, but not all combinations of beta and gamma subtypes can form dimers. For example, the gamma 2 subunit can form dimers with beta 1 and beta 2, but gamma 1 forms dimers only with beta 1, not with beta 2. Selective dimerization may play a role in the regulation of beta gamma dimer-mediated signal transduction. In order to identify the region of gamma responsible for selective dimer formation, a series of gamma 1/gamma 2 chimeras was constructed, transcribed, and translated in vitro. The ability of these gamma chimeras to form dimers with beta 1 and beta 2 was assayed by trypsin protection and chemical cross-linking. When amino acids 36-49 of gamma 1 were substituted for 33-46 of gamma 2, the chimera behaved like gamma 1 and dimerized only with beta 1; the reciprocal chimera, in which 14 residues from gamma 2 were substituted for the corresponding amino acids of gamma 1, behaved like gamma 2 and interacted with both beta 1 and beta 2. This 14-amino acid region was sufficient for gamma 1 to discriminate between the beta subunits. All gamma chimeras were functional because they were able to interact with beta 1, which is capable of forming dimers with both gamma 1 and gamma 2. All dimers of chimeric gamma subunits plus beta 1 were able to interact with purified alpha o subunit, indicating that beta gamma dimers containing chimeric gamma molecules were capable of interacting with an appropriate third molecule. This lays the foundation for using these gamma chimeras to study selective dimer interactions with various effectors and receptors.
Groups of 3-week-old specific pathogen-free chickens immunized with a commercial live-virus quail pox vaccine (Bio-Pox Q) were not protected against challenge with "variant" poxviruses isolated from chickens that were previously vaccinated with commercial fowl pox vaccine. The percentages of vaccinated chickens resistant to challenge with each of the five variant field isolates were 0%, 20%, 0%, 20%, and 10%, respectively. However, when immunity engendered by the variant field isolates was challenged with the commercial quail pox vaccine virus, 80%, 70%, 80%, 50%, and 60% of the vaccinates, respectively, were protected. Results from cross-immunity studies indicate that the commercial quail pox vaccine does share some immunologic relationship with these variant poxvirus field isolates, but not enough to be used in the control of some outbreaks of pox caused by variant poxviruses.
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6-Methylsalicylic acid synthase, the multifunctional enzyme complex that catalyzes the biosynthesis of the tetraketide 6-methylsalicylic acid, was modified by thiol-specific inhibitors and cross-linking reagents. Treatment with 1,3-dibromopropan-2-one caused rapid enzyme inactivation and formation of cross-linked dimers. Analysis by SDS-PAGE, density gradient ultracentrifugation, and secondary modification with [14C]iodoacetamide showed that two types of cross-linked dimers were formed. Peptides derived from native and 1,3-dibromopropan[2-14C]one-treated enzyme were isolated by SDS-PAGE and N-terminally sequenced. The sequences of the two N-termini from cross-linked peptides were located in the nucleotide-derived amino acid sequence and found to arise from the beta-ketoacyl synthase and acyl carrier protein components of the 6-methylsalicylic acid synthase subunit. Acetyl-CoA protected the enzyme from both inactivation and cross-linking by binding to the reactive cysteine of the beta-ketoacyl synthase component. Malonyl-CoA protected against cross-linking by binding to the thiol moiety of the 4'-phosphopantetheine prosthetic group of the acyl carrier protein. Formation of a mixed disulfide on treatment with 5,5'-dithiobis(2-nitrobenzoic acid) indicated that these two types of thiol residue are positioned close to each other in the active enzyme. From these studies, it was concluded that two pairs of functional dimers are present in the 6-methylsalicylic acid synthase tetramer and that, within each dimer, the beta-ketoacyl synthase and acyl carrier protein components are juxtaposed to allow the respective cysteine (residue 204) and 4'-phosphopantetheine thiols to interact during condensation. This spatial arrangement of thiols at the condensing active site is analogous to that found in type I vertebrate fatty acid synthases and other polyketide synthases.
PURPOSE: To evaluate the effect of coenzyme Q10 in reducing the skeletal muscle reperfusion injury following clamping and declamping the abdominal aorta. METHODS: 30 patients undergoing elective vascular surgery for abdominal aortic aneurysm or obstructive aorto-iliac disease were randomly divided into two groups: patients in group I were treated with coenzyme Q10 (150 mg/day) for seven days before operation, and those in group II received a placebo. We studied the hemodynamic profile in each patient during clamping and declamping of the abdominal aorta. The plasma concentrations of thiobarbituric acid reactive substances (malondialdhehyde), conjugated dienes, creatine kinase and lactate dehydrogenase were measured in samples from both arterial and inferior vena cava sites. Serial sampling was performed after induction of anesthesia, 5 and 30 minutes after abdominal aortic cross clamping, 5 and 30 minutes after aortic cross-clamp removal. RESULTS: The concentrations of malondialdehyde, conjugated dienes, creatine kinase and lactate dehydrogenase in patients who received CoQ10 were significantly lower than in the placebo group. Decrease of plasma malondialdehyde concentrations correlated positively (p < 0.01) with decrease of both creatine kinase and lactate dehydrogenase release in samples from the inferior vena cava. The hemodynamic profile during clamping and declamping the abdominal aorta was similar in both groups. CONCLUSIONS: Our findings suggest that pre-treatment with coenzyme Q10 may play a protective role during routine vascular procedures requiring abdominal aortic cross clamping by attenuating the degree of peroxidative damage.
Understanding how mammals satisfy their need for space in fragmenting ecosystems is crucial for ecosystem conservation. Using state-of-the-art global positioning system (GPS) technology we tracked 11 focal African elephants (Loxodonta africana) in Kenya at 3-hourly fix intervals and collected between 34 and 406 days per individual. Our recordings gave a high spatio-temporal resolution compared to previous studies and allowed novel insights into range use. The actual ranges of the tracked elephants are smaller than usually represented. Moreover, the ranges in our sample were complex and not confined to officially designated protected areas, except where fenced. All the unfenced elephants in our sample had distinct 'home sectors' linked by 'travel' corridors. Within each home sector the elephants concentrated in favourite 'core zones'. Such core zones tended to lie in protected areas whereas corridors typically crossed unprotected range. Elephants moved significantly faster along corridors than elsewhere in their range, which suggests awareness of danger outside the protected area. We conclude that understanding the complex use of an animal's range is crucial for conservation planning aiming to balance animal interests with those of human beings that co-habit in their range.
Pasteurellosis was induced in rabbits by conjunctival inoculation with 2 strains of Pasteurella multocida. The LD50 of strain P1062 (a bovine isolate) was 10(5.1) colony-forming units and that of strain P1059 (a turkey isolate) was 10(5.5) colony-forming units. Pasteurella-free rabbits were vaccinated IV or mucosally with boiled cells of P multocida or a cross-reactive uridine diphosphogalactose epimerase-deficient mutant of Escherichia coli J5. In rabbits challenge exposed with P multocida strain 1062 or 1059, homologous P multocida strain gave the best protection against fatal bacteremia. Partial protection was provided by J5; mucosal routes of vaccination (aerosol or conjunctival) gave better protection than did the IV route. Serum antibody titers were lower in rabbits vaccinated by mucosal routes than in those vaccinated IV. Cross-reactive IgG and IgM titers to P multocida were demonstrated when rabbits were vaccinated with J5. On the basis of bacteriologic examination of nasal secretions, rabbits that died were considered culture positive sooner than were those that survived. On the basis of bacteriologic examination of blood, rabbits that died were considered culture positive, and those that survived were considered culture negative. Seemingly, heat-stable antigens were protective, the cross-reactive E coli J5 mutant (with only core lipopolysaccharide) provided partial protection against pasteurellosis, and the mucosal route was somewhat useful for cross-protective immunization.
High levels of protection can be attained against Haemonchus contortus challenge infection in sheep using native antigens isolated from the gut of the adult parasite. However, vaccination with recombinant forms of these antigens, or components thereof, has disappointingly failed to generate similar levels of protection, suggesting that appropriate nematode glycosylation may be a prerequisite for protection. The free-living nematode, Caenorhabditis elegans is closely related to H. contortus and has been shown to share similar glycan moieties. In order to investigate the potentially protective role of these glycan moieties, a complex set of glycoproteins was isolated from C. elegans using ConA-lectin chromatography and their efficacy as immunogens against H. contortus challenge infection evaluated in sheep. Despite the generation of a high titre systemic IgG antibody response to the C. elegans glycoproteins and the ability of these antibodies to bind to the microvillar surface of the gut of H. contortus, no protection against challenge infection was observed. Serum antibodies to the C. elegans glycoproteins cross-reacted with the H. contortus host-protective antigen, H-gal-GP, by ELISA, although the level of cross-reactivity was not of a magnitude considered protective. Qualitative differences were also determined between the glycan epitopes of the C. elegans ConA-binding proteins and those of H-gal-GP, suggesting the presence of H. contortus-specific patterns of glycosylation.
Oxidative modification of the eye lens proteins, the crystallins, is known to cause protein cross-links and aggregates which lead to lens opacification or cataracts. We focus attention here on oxidative damage occurring in crystallins and some "control" proteins upon reaction with the hydroxyl radical (.OH) which, in the lens, is generated by photosensitization or by the Fenton reaction. In the present study, we have synthesized and used the bishydroperoxide I as a "photo-Fenton" reagent, in order to photolytically generate pure .OH, free of other oxyradicals. Our findings are the following: (i) Trp residues are oxidized by .OH to N-formylkynurenine and related compounds, but this in itself does not lead to covalent aggregation of the protein. (ii) Tyr residues react with .OH, but apparently do not produce dihydroxyphenylalanine or bityrosine. Nor do protein cross-links occur as a result. (iii) Oxidation of His residues appears to be obligatory for protein cross-linking. Histidine-free proteins do not form high molecular weight products upon reaction with .OH. Protection of His residues by adduct formation in other proteins inhibits cross-linking. (iv) Lys residues seem to participate in the cross-linking reaction. Protection of the Lys residues by maleylation of the protein inhibits cross-linking. (v) The oxidized protein is more acidic in nature than the parent, and it might have altered conformational features.
The potential usefulness of somatosensory evoked potential monitoring during aortic cross-clamping is slowly being realized. In addition, the protection of endangered spinal nervous tissue during aortic cross-clamping has not been sufficiently evaluated. To test the pharmacologic protective efficacy of various agents, we recorded spinal evoked somatosensory potentials (bipolar epidural catheter) in dogs under controlled conditions (N2O/O2-enflurane anesthesia) following clamping of the aorta for 1 hour. There were 5 groups of animals: those treated with different medications, such as prostaglandin E1 (PGE1), prostacyclin (PGI2), superoxide dismutase (SOD), and PGE1 plus SOD for pharmacologic protection during ischemia, and the controls. The time to recovery of evoked potentials during the reperfusion period was 36 minutes in the controls, 15.9 minutes in the SOD group (p < 0.01), 12.5 minutes in the PGE1 group (p < 0.001), 10.8 minutes in the PGI2 group (p < 0.001), and 3.8 minutes in the combination group (p < 0.001). In addition, treatment resulted in a better neurologic outcome on the seventh postoperative day when compared with the control group. While in the control group only 1 animal could walk (9%), 7 of 12 in the PGE1 group (58%), 4 of 12 in the SOD group (33.8%), 8 of 12 in the PGI2 group (66.7%), and all animals in the combination group (100%) could walk. We computed an exponential correlation that related the mean time of potential recovery during reperfusion with Tarlov scoring (grade 0 = paraplegia; grade 1 = paraplegia with little movements; grade 2 = paraparesis; grade 3 = paraparesis with some problems; grade 4 = normal motor function) in the various groups.(ABSTRACT TRUNCATED AT 250 WORDS)
The ciliate Ichthyophthirius multifiliis is an economically important parasite in aquaculture as well as in the ornamental fish trade. The parasite has been well studied and details of life cycle, host response and pathogenesis have been characterized. Parasite development within host epithelial tissues initiates localized leukocytic infiltrations, although the relationship between these responses and host resistance is uncertain, and whether or not leukocyte responses play a role in protective immunity is unclear. Here Frank Cross outlines the character of localized cellular responses during I. multifiliis infection and discusses the contribution of such responses to protective immunity and pathogenesis.
INTRODUCTION: Carotid angioplasty with stenting (CAS) is becoming accepted as an effective and reliable treatment option for severe carotid artery stenosis. However, it is rarely applied for carotid occlusion, especially in its chronic stage. We report our experience of CAS for chronic internal carotid artery occlusion representing compromised cerebral blood flow using various protection methods. METHODS: A 77-year-old woman, who was already diagnosed with severe left internal carotid artery stenosis, suddenly had right hemiparesis and aphasia. At that time, she was treated conservatively because her neurological status was quite good, in spite of left carotid artery occlusion. Her symptoms improved in the short term, except slight aphasia, but deteriorated again 18 days from the onset, and magnetic resonance imaging (MRI) showed new ischemic lesions. CAS was then performed for the occluded carotid artery on the 23rd day from the first onset. RESULTS: Using the proximal protection technique, the occluded lesion was crossed carefully with a microguidewire. Stents were also placed successfully with the distal protection technique. The occluded carotid artery was completely recanalized without any unfavorable events or neurological deterioration. CONCLUSION: In this patient, CAS was successfully to treat chronic carotid artery occlusion. These procedures and techniques are reviewed and discussed.
RAST-inhibition studies were performed by use of whole venom sac extracts to inhibit binding to purified venom allergens from various vespid wasps. Immunodiffusion studies were also performed with rabbit antisera raised against purified venom proteins. Immunologic cross-reactivity was demonstrated among the hyaluronidases, phospholipases, and antigen 5 s from both subgenera of yellow jackets, white-faced hornets, and paper wasps. The paper wasp hyaluronidase and antigen 5 were less related to those of the other three species than those of yellow jackets and hornets. It appears that immunologic cross-reactivity is the major mechanism of multiple allergic sensitivity to vespid venoms. Therapy with only the primary venom should be sufficient to protect against reactions from cross-reacting venoms.
Much evidence of immunogenic heterogeneity among the LC strains of Mycoplasma mycoides ssp. mycoides emerged from cross-immunization and -hyper-immunization experiments in mice in which three LC strains (Vom/Plum Island, 74/2488, and Mankefår 2833) were used for challenge purposes. All heterologous LC-strain vaccines cross-immunized against the three challenge strains, but protection was usually only 'partial', i.e. significantly less than that given by homologous vaccine. Cross-hyperimmunization with all heterologous LC but not SC strains produced protection against challenge with Vom/Plum Island that was virtually 'complete', i.e. similar to that produced by homologous vaccine. Challenge with 74/2488 gave generally similar results; but against Mankefår 2833 six heterologous LC vaccines gave complete protection and six did not. Vaccines prepared from the Smith (1423) strain of M. mycoides ssp. capri gave some protection against Vom/Plum Island but none against 74/2488 or Mankefår 2833. The cross-immunizing ability of three further M. mycoides ssp. capri strains appeared to resemble that of Smith (1423). In a cross-hyperimmunization experiment, vaccines prepared from SC strains of M. mycoides ssp. mycoides varied greatly in their ability to protect against challenge with strains 74/2488 and Mankefår 2833.
The majority of protein antigens of the malaria parasite Plasmodium falciparum contain short sequences that are extensively repeated in tandem arrays. Some antigens contain a single block of repeats whereas in other antigens there may be two or more blocks of related repeats. The repetitive sequences in an individual antigen may be highly conserved but more usually there is some degeneracy which occasionally is extensive. The repetitive sequences encode immunodominant epitopes to which much of the antibody response in malaria is directed. Recently, we have found that there are extensive cross-reactions amongst the epitopes encoded by related repetitive sequences. These cross-reactions may involve different blocks of repeats in the one antigen or repetitive sequences in different antigens. It is proposed that these cross-reactions interfere with the normal maturation of a high affinity antibody response in malaria by causing an abnormally high proportion of somatically-mutated B cells to be preserved during clonal expansion.
Previous studies have shown that the amino-terminal regions of group A streptococcal M proteins contain primarily protective (opsonic) epitopes and not tissue-cross-reactive epitopes. Limited primary structures from multiple serotypes of M protein containing only protective epitopes could potentially be linked together to form a broadly protective vaccine. The present studies were undertaken to determine the protective immunogenicity of a recombinant, multivalent hybrid molecule containing amino-terminal subunits of types 24, 5, 6, and 19 M proteins. Polymerase chain reaction primers were designed to amplify emm gene fragments ranging from 35 to 113 codons. The PCR products were ligated in tandem and inserted into pKK223-3. The tetravalent M protein that was purified from extracts of Escherichia coli migrated as a single band on SDS-PAGE with an apparent m.w. of 31 kDa. In immunoblot analyses, the hybrid protein reacted with serotype-specific antisera indicating that it contained all four M protein subunits. Rabbits immunized with the purified tetravalent M protein developed significant antibody levels against all four serotypes of native M proteins represented in the hybrid protein. None of the antisera cross-reacted with human tissues. The immune sera also opsonized all four serotypes of group A streptococci. Our data show that a hybrid protein containing subunits from multiple M proteins can evoke broadly protective immune responses without tissue-cross-reactive antibodies.