Iodine content of rat thyroglobulin affects its antigenicity in inducing lymphocytic thyroiditis in the BB/Wor rat. The BB/Wor rat develops spontaneous insulin dependent diabetes mellitus (DM) and lymphocytic thyroiditis (LT). We have recently demonstrated that immunization of BB/Wor rats with allogeneic thyroglobulin (Tg) induces LT at an early age. The incidence of spontaneous and Tg induced LT is extremely variable among different BB/Wor sublines. It has been shown that high iodine diet significantly increases the incidence of spontaneous lymphocytic thyroiditis (LT) and low iodine diet significantly decreases the incidence of LT in genetically predisposed BB/Wor rats. Recent studies on thyroglobulin (Tg) induced LT in chicken and mouse have shown that iodine rich Tg is far more antigenic than Tg with a low iodine content, suggesting that a high iodine diet increases the immunogenicity of Tg molecule. In order to determine whether the extent of Tg iodination would affect its immunogenicity in the BB/Wor rats, the current study was carried out. Normal iodine Tg (NTg) or low iodine Tg (LTg) was obtained from thyroids of rats that were placed on regular diet or regular diet plus 0.5% methimazole, respectively. 120 rats from the NB (highly susceptible) and BB (low susceptible) sublines were randomized in three groups. Immunization was carried out with a 1:1 emulsion of complete Freund's adjuvant (CFA) and LTg, NTg (0.6 mg/rat) or saline at 30 and 37 days of age. Since spontaneous LT rarely occurs before age 75 days, rats were sacrificed at age 65 days to specifically study Tg induced LT. Immunization with NTg induced LT in 31% of the NB rats, but not in the BB subline. LTg did not induce LT in either subline.(ABSTRACT TRUNCATED AT 250 WORDS)


Autoantibodies to c-myc protein: elevated levels in patients with African Burkitt's lymphoma and normal Ghanians. Sera from U.S. patients with SLE, RA, and various malignancies, clinically normal individuals with sero-activity to HIV, AIDS, and from pregnant women were tested for the presence of anti-c-myc antibodies. In an ELISA using recombinant human c-myc protein as the antigen, no difference in mean antibody titer was generally detected in these sera when compared to normal controls. Only three malignancy sera (two myeloid leukemia and only one lymphoma) and two patients with AIDS-related lymphoma exhibited exceedingly higher levels of anti-c-myc antibody. However, significantly elevated anti-c-myc antibody levels were found among 20 patients with African Burkitt's lymphoma (Ghana) and 20 normal Ghanians, thus apparently reflecting an autoimmune phenomenon prevalent in the endemic region. These findings indicated that elevated levels of anti-c-myc antibodies are not a general characteristic of patients with diseases that have been associated with increased expression of c-myc.


The nuclear autoimmune antigen Ku is also present on the cell surface. Polyclonal antibodies were raised against the individual 85 and 70 kDa subunits of the Ku complex purified from nuclear extract prepared from the T cell line MLA144. They specifically recognise the appropriate subunits of the Ku complex from whole cell extract of HeLa cells using Western blot analysis. They are also able to identify the Ku proteins present in the cell membrane using FACS analysis.


Probe-less genomic typing of Arg52 (type 1 diabetes-associated) and non-Arg52 (non-type 1 diabetes-associated) HLA-DQA1 alleles. According to recent evidence, the presence of an Arg residue at position 52 in the HLA-DQ alpha chain may confer susceptibility to Type 1 diabetes and thus be possibly used to define quantitatively the genetic risk of this disease. Arg52 and non-Arg52 DQA1 alleles cannot be typed by the conventional cytotoxicity test and they must be distinguished at the genomic level. We describe a simple procedure which discriminates the DQA1 alleles based on the differential electrophoretic migration of the DNA heteroduplexes they form with a reference DNA fragment. A major advantage of this procedure is the fact that no hybridization probe is required. Practically, this typing procedure consists of an electrophoretic run of the products of a selective PCR in polyacrylamide gel.


Reversal of the abnormal development of T cell subpopulations in the thymus of autoimmune MRL-lpr/lpr mice by a polyamine biosynthesis inhibitor. Polyamines--putrescine, spermidine, and spermine--are a group of positively charged organic molecules that are present in all living cells. They are important regulators of cell growth and differentiation, but the precise mechanism of their action is not known. Ornithine decarboxylase (ODC) is a key enzyme in the biosynthesis of polyamines. Recent studies demonstrated that down-regulation of polyamine biosynthesis by irreversible inhibition of ODC with difluoromethylornithine (DFMO0 is a novel therapeutic approach for the treatment of murine lupus in autoimmune MRL-lpr/lpr mice. Since murine lupus in this strain is associated with a major alteration in thymic T cell subopulations, we questioned whether abnormal polyamine biosynthesis contributes to aberrant T cell maturation in the thymus of MRL-lpr/lpr mice. Thymocytes were analyzed for cell surface markers, CD4 and CD8 by 2-color flow cytometry using their respective monoclonal antibodies. The proportion of thymocyte subsets in disease-free mice (8-10 week of age) was approximately 72% double positive (DP; CD4+CD8+) cells, 5-7% double negative (DN; CD4-CD8-) cells, 11-16% CD4+ cells and 7-8% CD8+ cells. At 14 weeks of age, a stage of clinical disease expression, thymocytes were marked by the presence of approximately 40% DN cells and approximately 25% DP cells.(ABSTRACT TRUNCATED AT 250 WORDS)


Does mitogen-induced antibody production by normal blood cells mimic spontaneous production in lupus? Blood cells from patients with systemic lupus erythematosus (SLE) showed a raised level of spontaneous IgG production that included antibodies to DNA and to common environmental antigens (influenza virus haemagglutinin, adenovirus hexon and mannan from Candida albicans). In contrast, no IgG antibody was produced against an antigen not normally encountered in the UK (egg antigen from Schistosoma mansoni) or a self-antigen not generally associated with SLE (thyroglobulin). IgM production was raised to a lesser extent and only antibodies to DNA were detected. When normal cells were stimulated with pokeweed mitogen or S. aureus organisms, the specificity pattern of IgG production was similar to that described above for SLE with the major exception of the absence of IgG anti-DNA. IgM antibodies to DNA and all the other antigens were detected, but the specificity of the IgM ELISA assays for the protein antigens needs further clarification. The activity of IgM anti-DNA relative to total IgM was far greater in the SLE system. These results provide further evidence that a response to self-antigen is required for production of pathogenic IgG autoantibodies in SLE.


Rabbits produce SLE-like anti-RNA polymerase I and anti-DNA autoantibodies in responses to immunization with either human or murine SLE anti-DNA antibodies. Anti-DNA and anti-DNA polymerase I (RPI) autoantibody responses are symptoms of systemic lupus erythematosus (SLE). To investigate the relationship between these antibodies (Ab), rabbits were immunized with one of the following preparations: human SLE anti-DNA Ab; human SLE anti-DNA IgG; normal human anti-DNA Ab; human Grave's disease anti-DNA Ab; murine SLE anti-DNA Ab or anti-DNA IgG Fab; various normal human, murine, or rabbit IgG preparations; or complete Freund's adjuvant (CFA), alone. All of the animals immunized with anti-DNA Ab (n = 14) generated Ab reactive in radioimmunoassay with: ssDNA, dsDNA, RPI, the soluble fraction of rabbit liver crude nuclear extract, and the immunogen. Induced rabbit anti-DNA Ab in turn induced these responses in a different rabbit: a rabbit immunized with rabbit anti-DNA IgG Ab which had been previously induced by immunization with human anti-DNA Ab, produced Ab reactive with ssDNA, dsDNA, RPI, and the soluble fraction of rabbit liver nuclear extract. Although an individual animal's antisera reacted consistently over the course of immunization with the same individual RPI subunit(s), antisera from different animals reacted with different subunits of the 9-subunit RPI complex in Western blot analyses: 190 kD (n = 6); 120 kD (n = 1); 62 kD (n = 4); 45 kD (n = 2); and, no reactivity (n = 2). In contrast, animals immunized with normal IgG or CFA produced responses only against the immunogen. Together, these data suggest that anti-DNA and anti-RPI responses are connected through an autoimmune network in SLE.


Immunopathology of experimental autoimmune uveoretinitis in primates. The eyes and pineal glands from 10 monkeys immunized with S-antigen were studied using routine histopathological and immunohistochemical techniques. Seven out of 10 animals developed uveitis between 19 and 33 days after the initial immunization. Histopathology of the eyes harvested 70 days after immunization showed moderate to marked uveoretinitis, subretinal fibrosis, retinal necrosis and gliosis. The pineal glands demonstrated chronic pinealitis. The infiltrating cells were both CD3 and CD19/CD22 lymphocytes with a ratio of 1.4 in the eye and 2.2 in the pineal gland. The ratio of CD4 to CD8 lymphocytes was 1.5:1. MHC Class II antigens and adhesion molecule (ICAM-1) were observed on resident cells. The influx of B lymphocytes and the formation of subretinal fibrosis differentiate the disease in the monkey from that in the rat and mouse. These findings are similar to Vogt-Koyanagi-Harada syndrome and subretinal fibrosis with uveitis syndrome in human.


Cellular immune mechanisms in chronic autoimmune thrombocytopenic purpura (ATP). Chronic autoimmune thrombocytopenic purpura (ATP) is a common autoimmune-mediated bleeding disease in which autoantibodies are directed against platelets, resulting in their enhanced Fc-mediated destruction by macrophages in the spleen. While there has been extensive studies relating to the autoantibodies in this autoimmune disorder, relatively few have dealt with cell-mediated immunoregulation of the anti-platelet autoantibody response. Nonetheless, there is accumulating evidence that suggests the production of these anti-platelet autoantibodies is under the influence of several abnormal lymphocyte-mediated mechanisms, i.e. enhanced anti-platelet T helper cell activity with concomitant reduced T suppressor cell activity. This review focuses on these cellular events and presents a working model which attempts to explain their close interrelationships.


Mediators of joint swelling and damage in rheumatoid arthritis and pristane induced arthritis. Joint swelling and tenderness in rheumatoid arthritis (RA) probably result from IgG aggregates activating complement with the consequent attraction of polymorphonuclear leucocytes (PMNs) and the liberation of their granule enzymes such as kininogenases. By contrast IL-1 and TNF are the major stimulants of cartilage and bone loss although other agents contribute. The fundamental drive for the production of these mediators is unknown but a role for heat shock proteins is suggested from work on pristane induced arthritis.


Postpartum thyroiditis: an organ specific syndrome which is not associated with a postpartum polyclonal B-cell activation. Recent reports have detailed the presence of autoantibodies characteristic of non-organ specific autoimmune diseases in the serum of patients with autoimmune postpartum thyroiditis (PPT). These observations suggest that PPT could be part of a polyclonal B-cell activation postpartum. We have measured 4 non-organ specific autoantibodies (anti-DNA, anti-cardiolipin, anti-nuclear factor (ANF) and antibodies against extractable nuclear antigens (ENA)) together with autoantibodies against thyroglobulin and thyroid peroxidase in a group of PPT women at 4 time points during the first year postpartum (early, time of hyperthyroidism, time of hypothyroidism, late). Whilst 18/18 patients showed thyroid specific autoantibody changes there was only a low frequency of occurrence of the non-organ specific autoantibodies (ENA 2/18; ANF 1/18; anti-DNA 2/18; anti-dsDNA 0/18; anti-cardiolipin 0/18) and, when positive, the response was poor. We conclude that PPT is not associated with a polyclonal b-cell activation but is a postpartum rebound of a thyroid specific autoimmune phenomenon.


Conformational properties of oxytocin in dimethyl sulfoxide solution: NMR and restrained molecular dynamics studies. The conformation of oxytocin, the neurohypophyseal nonapeptide hormone, in solution in deuterated dimethyl sulfoxide has been determined by 1H-nmr. The structural determination is based on the experimental data set of nuclear Overhauser effect restraints. Obtained after the restrained molecular dynamics simulation on an initial structure of extended conformation, five resultant structures satisfy the experimental restraints well. These structures resemble that of the crystal structure of deamino-oxytocin, an analogue of oxytocin, in terms of a close correlation observed both at two beta-turn regions of the 20-membered tocin ring and at the tripeptide tail end. Based on this comparison and analysis of restrained molecular dynamics trajectories, we found that, although the turns are stabilized by the formation of hydrogen bonds, the oxytocin molecule possesses a slight twist in DMSO solution relative to the orientation of deamino-oxytocin in the crystalline state. Analyses of oxytocin conformation indicate that the tripeptide tail is more flexible than the tocin ring.


Amino acid side-chain populations in aqueous and saline solution: bis-penicillamine enkephalin. The potentials of mean force (pmfs) for rotation around the chi 1 aromatic side-chain dihedrals of the zwitterionic bis-penicillamine enkephalin pentapeptide have been determined in both aqueous and saline solution. These side chains are known to be associated with the pharmacophore and their conformational populations are thought to be critical for activity. It is found that the association between chloride ions and the peptide in saline solution simulations has profound effects on the relative energies of the g-, g+, and t conformations, and also the barriers between them. Using the pmfs we have also calculated the respective Boltzmann-weighted 3J alpha beta vicinal coupling constants. The agreement between the calculated and experimentally determined coupling constants is poor for the pmf in pure water, but substantially improved for the pmf determined in saline solution. Reasons for these differences appear to be related to the experimental conditions.


A study of the quadrupolar NMR splittings of 7Li+, 23Na+, and 133Cs+ counterions in macroscopically oriented DNA fibers. The hydration and temperature dependencies of the 23Na+, 133Cs+, and 7Li+ quadrupolar splitting have been determined in hydrated, macroscopically oriented DNA fibers. At low water contents the quadrupolar splitting is found to decrease as the water content increases, regardless of counterion, while at high water contents the hydration dependence is reversed. The 23Na+ and 133Cs+ quadrupolar splittings decrease as the temperature increases, while the 7Li+ splitting shows the opposite behavior. At high water contents the 23Na+ and 133Cs+ splittings decrease, and then, after passing zero splitting, increase as the temperature increases. The interpretation of the temperature dependence is discussed in terms of a two-site model (free and bound ions) and a three-site model (free ions and specifically or nonspecifically bound ions). It is suggested that a three-site model is more consistent with the data for the present system. At high water contents, the temperature dependence of the 7Li+ splitting vanishes, indicating counterion condensation. The behavior of the 7Li+ splitting is confirmed by measurements on DNA fibers in equilibrium with a C2H5OD-D2O-LiCl solution. The salt dependence in this system is weak. The counterion quadrupolar splitting is seen to be very sensitive to structural transitions in double-helical DNA.


Viscoelastic behavior of fractionated ovine submaxillary mucins. Solution properties of fractionated ovine submaxillary mucin (OSM) and asialo OSM (aOSM) in aqueous guanidine hydrochloride have been investigated using light scattering and rheological methods. For the first time we present viscometric evidence in both dilute and concentrated solution that the molecular structure of OSM is that of a wormlike chain. The intrinsic viscosity shows molecular weight dependence consistent with the linear extended chain conformation observed by light scattering measurements. The viscoelastic behavior of the OSM fractions in aqueous guanidine hydrochloride was further examined above the overlap concentration as a function of molecular weight and temperature. Under these solvent conditions in which the role of nonbonding intermolecular interactions is minimized, OSM shows predominantly fluid like behavior. However, high molecular weight OSM shows evidence of the existence of an entanglement network at high concentration. The frequency-dependent shear storage and loss moduli at all concentrations and molecular weights can be scaled to yield a master curve by incorporating typical viscoelastic shift parameters. The entanglement molecular weight and concentration are consistent with literature data for extended, semiflexible wormlike chains. The behavior of aOSM is similar to that of intact OSM at comparable degrees of coil overlap, indicating that the terminal sialic acid residue on the carbohydrate side chain has no effect on the rheology of concentrated OSM solutions beyond that due to an increase in the hydrodynamic volume.


Alpha-helix to random coil transitions: determination of peptide concentration from the CD at the isodichroic point. A method is presented for determining the concentrations of peptides and proteins having isodichroic points near 203 nm. The existence of an isodichroic point for a given substance indicates a local two-state (alpha-helix, random coil) population. The mean residue ellipticity at the isodichroic point, [theta lambda i], is, of course, independent of helix content. For a wide variety of synthetic and natural peptides, including both single helices and coiled coils, it is shown that [theta lambda i] is also essentially independent of substance and of whether the transition is induced by temperature, ionic strength, pH, chain length changes, amino acid substitution, or solvent perturbation. Averaging [theta lambda i] values culled from various laboratories gives -151 +/- 16 (SD, 7 sources) deg.cm2.mmol-1. In our laboratory, nonpolymerizable rabbit alpha-tropomyosin and two alpha-tropomyosin subsequences yield -135 +/- 10 (SD, 190 values) deg.cm2.mmol-1. Thus, given [theta lambda i] for a peptide of known concentration, it is possible to estimate the concentration of any other peptide provided that it has an isodichroic point at which the ellipticity is accurately measurable. It is then possible to calculate [theta lambda] at any other wavelength for which theta is known. It is advisable to determine [theta lambda i] for the best known peptide in one's own laboratory, since it depends on absolute instrument and cell calibrations and an absolute concentration determination.


How many numbers are required to specify sequence-dependent properties of polynucleotides? There are 10 unique dinucleotides of double-stranded DNA, but only 8 independent nearest-neighbor energies that occur in circular DNA, as shown by D. M. Gray and I. Tinoco [(1970) Biopolymers 9, 223-244]. We extend that analysis to include end effects, and show that the number of unique dinucleotide pairs (including ends) is 14, but there are only 12 independent energies. We discuss how these 12 energies (or spectra or any other pairwise additive property) can be measured and displayed, and how they should and should not be compared between experimenters. As an example, we analyzed the recently reported melting curves [M.J. Doktycz et al. (1992) Biopolymers, 32, 849-864.] of 16 DNA dumbbells in two different Na+ environments. This analysis reveals a new means for evaluating end effects and the emergence of longer than nearest-neighbor interactions at low salt concentration.


Helix propagation in trifluoroethanol solutions. Helix propagation of the S-peptide sequence (residues 1-19 of ribonuclease A) in 2,2,2-trifluoroethanol (TFE) solutions has been investigated with CD and nmr Overhauser effect spectroscopies. In this study, the S-peptide helix is covalently initiated at the N-terminus through disulfide bonds to a helix scaffold derived from the N-terminal sequence of the bee venom peptide apamin. The entire S-peptide sequence of this hybrid sequence peptide becomes helical at high proportions of TFE. Residues 14-19 of the S-peptide are not helical in the free peptide in TFE, nor are they helical in ribonuclease A. The "helix stop" signal encoded by the S-peptide sequence near residue 13 does not persist at high TFE with this hybrid sequence peptide. The helix-stabilizing effects of TFE are due at least in part to facilitated propagation of an extant helix. This stabilizing effect appears to be a general solvation effect and not due to specific interaction of the helical peptide with TFE. Specifically these data support the idea that TFE destabilizes the coil state by less effective hydrogen bonding of the peptide amide to the solvent.


Hexaamminecobalt(III) binding environments on double-helical DNA. Previous cation nmr evidence suggests that univalent cations such as Na+ bind to DNA in a diffuse, nonspecific manner, whereas di- and trivalent cations show distinct binding heterogeneity. Here are reported 59Co- and 23Na-nmr measurements of the %GC dependence of the DNA binding behavior of the trivalent hexaamminecobalt(III) cation. When Co(NH3)6Cl3 titrations are performed on one mammalian and three bacterial DNAs, evidence is found for at least three distinct classes of bound Co(NH3)6(3+). A comparison of titration curves for all four DNAs demonstrates that an increase in GC content correlates with an increase in the fraction of specific Co(NH3)6(3+). binding sites. For M. lysodeikticus DNA (72% GC), a slowly exchanging class of bound 59Co(NH3)6(3+) is apparent. This class of sites is saturated at very low binding densities (between 0.02 and 0.03 cobalt cations per DNA phosphate). At higher binding densities (greater than 0.03), the signal due to slowly exchanging 59Co(NH3)6(3+) disappears into the noise, and a single 59Co(NH3)6(3+) signal is observed. Within the sensitivity limitations of these measurements, no evidence for slowly exchanging bound 59Co(NH3)6(3+) could be found for any of the other DNAs, for which a single, rapidly exchanging 59Co(NH3)6(3+) signal is observed at all binding densities. For this rapidly exchanging signal, for all four DNAs, the measured 59Co(NH3)6(3+) nmr parameters depend significantly on (a) binding density and (b) GC content of the DNA.(ABSTRACT TRUNCATED AT 250 WORDS)


Solution conformations of two flexible cyclic pentapeptides: cyclo(Gly-Pro-D-Phe-Gly-Ala) and cyclo(Gly-Pro-D-Phe-Gly-Val). In an effort to explore the residue preferences in three-residue reverse turns (so-called gamma-turns), two cyclic pentapeptides--cyclo(Gly1-Pro2-D-Phe3-Gly4-Ala5) (I) and cyclo(Gly1-Pro2-D-Phe3-Gly4-Val5) (II)--have been synthesized and analyzed by nmr. It was anticipated that the Gly-Pro-D-Phe-Gly portions of these molecules would favor a beta-turn conformation, leaving the remainder of the molecule to adopt a gamma turn, as seen in several previously studied model cyclic pentapeptides. The nmr data for both peptides in CDCl3 (5% DMSO-d6) and in neat DMSO-d6 indicate that the most populated conformation contains a distorted beta turn around Pro2-D-Phe3, which includes a gamma turn around D-Phe3. The distortion in the beta turn does not impede the formation of an inverse gamma turn around residue 5, and indeed, this conformation is observed in both peptides. Both the alanine and the bulkier valine residues are therefore found to be compatible with an inverse gamma turn. Molecular dynamics simulations on the title peptides are reported in the following paper. These simulations indicate that there is conformational flexibility around the D-Phe3-Gly4 peptide bond, which enables the formation of the gamma turn around D-Phe3. The third paper in this series explores the impact of a micellar environment on conformational equilibria in II.


Combined use of molecular dynamics simulations and NMR to explore peptide bond isomerization and multiple intramolecular hydrogen-bonding possibilities in a cyclic pentapeptide, cyclo(Gly-Pro-D-Phe-Gly-Val). The conformational behavior of a model cyclic pentapeptide--cyclo(Gly-L-Pro-D-Phe-Gly-L-Val)--has been explored through the combined use of in vacuo molecular dynamics simulations and a range of nmr experiments (preceding paper). The molecular dynamics analysis suggests that, despite the conformational constraints imposed by formation of the pentapeptide cycle, this pentapeptide undergoes conformational transitions between various hydrogen-bonded conformations, characterized by low energy barriers. An inverse gamma turn with Pro in position i + 1 and a gamma turn with D-Phe in position i + 1 are two alternatives occurring frequently. Like other DLDDL cyclic pentapeptides, cyclo(Gly-Pro-D-Phe-Gly-Val) is also stabilized by an inverse gamma-turn structure with the beta-branched Val residue in position i + 1, and this hydrogen bond is retained in the different conformational families. The gamma-turn around D-Phe3 and the inverse gamma turn around Val5 are consistent with the nmr observations. 3JNH-CH alpha coupling constants of the all-trans forms were calculated from one of the molecular dynamics trajectories and are comparable to nmr experimental data, suggesting that the conformational states visited during the simulation are representative of the conformational distribution in solution. In addition to the equilibrium among various hydrogen-bonded all-trans conformers, the observation in nmr spectra of two sets of resonances for all peptide protons indicated a slow conformational interconversion of the Gly-Pro peptide bond between trans and cis isomers. The activation energy between these two conformers was determined experimentally by magnetization transfer and was calculated by high temperature constrained molecular dynamics simulation. Both methods yield a free energy of activation of ca. 20 kcal/mol. Furthermore, the free energy of activation is dependent on the direction of rotation of the Gly-Pro peptide bond.


Impact of a micellar environment on the conformations of two cyclic pentapeptides. In an effort to explore the influence of interfacial environments on reverse turns, we have performed a detailed analysis by nmr of the solution conformations of two cyclic pentapeptides in sodium dodecyl sulfate (SDS) micelles. The first peptide, cyclo (D-Phe1-Pro2-Gly3-D-Ala4-Pro5), adopts a single rigid conformation in solution (either chloroform or dimethylsulfoxide) and in crystals, whereas the second, cyclo (Gly1-Pro2-D-Phe3-Gly4-Val5), is much more flexible and adopts different conformations in the crystal and in solution. Both of these peptides are solubilized by SDS micelles, and nmr relaxation rates indicate that they are both partially immobilized by interaction with the micelles. Furthermore, some amide protons in both peptides participate in hydrogen bonds with water. In the presence of micelles, the former peptide retains a conformation essentially the same as that found in crystals and in solution, which consists of a beta turn and an inverse gamma turn. However, the micellar environment has a significant effect on the latter peptide. In particular, the population of a conformer containing a cis Gly-Pro peptide bond is increased significantly. The most likely conformation of the cis isomer, determined by a combination of nmr and restrained molecular dynamics, contains a Gly1-Pro2 delta turn and a gamma turn about D-Phe3. The nmr data on the trans isomer indicate that this isomer is averaging between two conformations that differ mainly in the orientation of the D-Phe3-Gly4 peptide bond.


Conformational analysis of Met-enkephalin in both aqueous solution and in the presence of sodium dodecyl sulfate micelles using multidimensional NMR and molecular modeling. Proton and 13C chemical shift assignments are reported for the neuropeptide Met-enkephalin (ME) in both aqueous solution and in the presence of 50 mM sodium dodecyl sulfate (SDS). Rotating frame nuclear Overhauser enhancement spectroscopy was used to qualitatively describe interproton distances. These distances were then used as restraints in the distance geometry based molecular modeling program Dspace, developed by Hare Research to generate sets of conformations of ME. The resulting aqueous solution conformations of ME were determined to exhibit characteristic of an extended random-coil polypeptide with no distinguishable secondary structure. The resulting set of solution conformations of ME in the presence of 50 mM SDS exhibited characteristics of an amphiphilic type IV beta turn that are stabilized by hydrophobic aromatic-aromatic interactions between the side chains of Tyr1 and Phe4.


QRS complex changes in the V5 ECG lead during cardiac surgery. The QRS complex in lead V5 was studied during cardiac surgery. R wave amplitude decreased after induction of anesthesia to approximately 50% to 60% of the preanesthetic level before the institution of CPB (P < 0.001). An rS complex appeared immediately after cardioversion and changed in configuration to an Rs complex 15 to 30 minutes after aortic declamping. The R wave continued to recover toward the preanesthetic level at sternal closure. Patients with coronary artery disease had a poorer recovery of the R wave (P < 0.05) than patients with valvular heart disease; the former recovered to only 50% of the preanesthetic level at sternal closure. Nonsurvivors had much smaller R waves (26.1 +/- 20.5%) than survivors (P < 0.001). The R wave peaked 30 to 40 ms after initiation of the QRS complex, which indicates recovery of conductivity and the activation sequence of the left ventricular (LV) free wall, which is easily disturbed by hypothermia, cardioplegia, and ischemia during aortic cross-clamping. Monitoring QRS complex changes in lead V5 appears to be important on weaning from cardiopulmonary bypass to detect regional ischemia, and also to observe electrophysiologic recovery of the LV free wall.


Temporary transmyocardial pacing using epicardial pacing wires and pacing pulmonary artery catheters. This study investigated the feasibility of transmyocardially pacing the heart using one temporary epicardial pacing lead and one endocardial lead of a pacing pulmonary artery catheter. Twenty patients undergoing cardiopulmonary bypass with cardioplegic arrest were studied 10 to 45 minutes and 18 to 30 hours after discontinuation of cardiopulmonary bypass. The Swan-Ganz Flow-Directed Pacing TD Catheter (Baxter Healthcare Corporation) was inserted in one group of 10 patients, and the Swan-Ganz Thermodilution A-V Paceport Catheter (Baxter Healthcare Corporation, Irvine, CA) was used in another group of 10 patients. Using the Pacing TD Catheter, transmyocardial atrial (TMA) pacing was successful in 14 of 16 attempts (87.5%), and transmyocardial ventricular (TMV) pacing was successful in 15 of 16 attempts (93.8%). With the AV Paceport Catheter, TMA pacing was successful in 16 of 18 attempts (88.9%), and TMV pacing was successful in 17 of 19 attempts (89.5%). Transmyocardial atrial-ventricular sequential pacing was achieved in all cases when both TMA and TMV pacing were independently successful. There were no significant differences between catheters in the success rates of either TMA or TMV. It is concluded that transmyocardial pacing is feasible using one temporary epicardial pacing lead and one endocardial lead of a pacing pulmonary artery catheter.


Continuous intra-arterial oximetry, pulse oximetry, and co-oximetry during cardiac surgery. This study evaluated arterial catheter oximetry versus pulse oximetry in eight patients (ASA III-IV) who underwent cardiac surgery. Co-oximeter saturation values served as the standard. Arterial oxygen saturation was determined simultaneously with these three methods at 162 prospectively defined points of measurement before, during, and after cardiopulmonary bypass (CPB). At the same times before and after CPB, arterial, pulmonary arterial, and central venous pressures, and cardiac output determinations were recorded. Saturation readings were obtained in more than 99% of measurements with catheter oximetry and in only 59% to 84% of measurements with pulse oximetry. Failure of pulse oximetry correlated with low mean arterial pressures and low cardiac outputs, but not with high systemic vascular resistance. The mean saturation values determined by catheter oximetry as well as by pulse oximetry differed from the mean values obtained by co-oximetry by less than 1% (= bias). The standard deviations of the individual differences between readings of catheter or pulse oximetry and readings of co-oximetry (= precision) were +/- 0.5% to +/- 1.0% for catheter oximetry and +/- 1.0% to +/- 1.2% for pulse oximetry. In summary, catheter oximetry was superior to pulse oximetry with regard to obtaining readings and to reliability of the obtained readings. Invasiveness and high costs influence the decision as to whether to use catheter oximetry, but if reliable and precise measurements of saturation are important at any time during surgery, pulse oximetry is an insufficient method and co-oximetry is a time-consuming method of analysis, whereas catheter oximetry is quick, reliable, and precise.


Repeated dose administration of desmopressin acetate in uncomplicated cardiac surgery: a prospective, blinded, randomized study. The effects of single or repeated doses of desmopressin on blood loss were examined in uncomplicated cardiac surgery, while assessing the potential for thrombogenic side effects. Seventy patients undergoing elective coronary artery bypass grafting (CABG) were studied. Patients were randomized into three blinded groups: Group I received DDAVP (0.3 micrograms/kg), IV, after cardiopulmonary bypass (CPB) and 12 hours later in the Intensive Care Unit (ICU); Group II, DDAVP (0.3 micrograms/kg), IV, after termination of CPB and saline (placebo) 12 hours later in the ICU; Group III, saline (placebo) IV after CPB and 12 hours later in the ICU. Blood loss and bleeding time decreased for Group I at 24 hours (P < 0.04) when compared to Group III; however, blood product replacement, as well as intraoperative and total blood loss at 36 hours, were not different among treatment and control groups. There were four myocardial infarctions recorded in Group I, two in Group II, and one in Group III. These differences were not found to be statistically significant. It is concluded that in routine CABG the prophylactic use of single or repeat dose DDAVP does not effectively decrease blood loss or blood product replacement.


Modest doses of nitroglycerin do not interfere with beef lung heparin anticoagulation in patients taking nitrates. The results of a prior clinical report suggested that nitroglycerin may interfere with the anticoagulant effect of heparin. Therefore, 30 adult patients undergoing cardiac surgery were studied in a controlled, prospective fashion. Thirteen patients on chronic nitrate therapy received an intraoperative nitroglycerin infusion at 1 micrograms/kg/min intravenously. Seventeen patients received no preoperative or intraoperative nitrates (control group). Heparin, 300 units/kg, was administered to all patients in three consecutive doses: 40 units/kg, 80 units/kg, and 180 units/kg. The activated coagulation time and activated partial thromboplastin time were measured prior to heparin, and 5 minutes after each heparin dose. There were no differences in automated activated coagulation times or in activated partial thromboplastin times between the groups at any measurement period. The study is limited in that only patients on chronic nitrates were included in the treatment group and that only a modest dose of nitroglycerin was used. However, it is concluded that a modest dose of intravenous nitroglycerin does not interfere with the anticoagulant effect of boluses of beef lung heparin in patients undergoing cardiac surgery.


Changes in plasma atrial natriuretic peptide concentration during heart transplantation. Examination of changes in plasma atrial natriuretic peptide (ANP) concentrations during heart transplantation may provide important information about factors influencing plasma ANP in patients with severe heart failure. Serial changes in plasma ANP during heart transplantation, and atrial content of ANP in native and donor atria, were measured in 12 patients. Preoperative plasma ANP was elevated in all patients (387 +/- 77 pg/mL), whereas atrial content of ANP in native atria was reduced (0.36 +/- 0.082 micrograms/mg protein). Preoperative plasma ANP did not correlate with hemodynamics, but was negatively correlated with creatinine clearance (r = -0.76, P < .01). Intraoperative plasma ANP prior to transplantation was strongly correlated with intraoperative plasma ANP after transplantation (r = 0.84, P < .001). Although postoperative plasma ANP was reduced from preoperative plasma ANP by 75%, these two measurements were also significantly correlated (r = 0.70, P < .02). Postoperative plasma ANP was not correlated with hemodynamics, but was negatively correlated with both creatinine clearance (r = -0.65, P < .05) and content of ANP in the native atria (r = -0.75, P < .01). Multiple linear regression analysis suggested that up to 85% of the variability of early postoperative plasma ANP could be accounted for by the variability in these latter two parameters. The decrease in native atrial ANP content, in the context of elevated plasma ANP concentration, is consistent with prior animal studies suggesting that severe heart failure induces cellular adaptations favoring accelerated ANP synthesis and secretion (with resultant reduction in tissue content).(ABSTRACT TRUNCATED AT 250 WORDS)


Variations of blood PAF-acether levels during coronary artery surgery. Extracorporeal circulation (ECC) is associated with thrombocytopenia and transient leukopenia. After ECC and coronary artery bypass graft (CABG) surgery, some patients can develop pulmonary and cardiac dysfunction, which might be related to the release of various mediators such as thromboxane A2, C5a, and C3a anaphylatoxins. The involvement of PAF-acether (PAF), a potent vasoactive thrombocytopenic and leukoneutropenic agent, has not been determined. Therefore, 10 patients were studied during and after CABG. The release of PAF, lipo PAF (PAF bound to blood lipoproteins), and lyso PAF (PAF precursor and metabolite) was measured in blood from the left atrium, radial artery, and pulmonary artery before and after CABG. PAF, lipo PAF, and lyso PAF were also determined during ECC at the entry and exit points of the oxygenator. Hemodynamic parameters, platelet, and leukocyte counts in the pulmonary artery were measured simultaneously. PAF did not increase significantly during ECC; it showed a transitory six-fold increase immediately after CABG in the radial artery (0.18 +/- 0.13 v 1.09 +/- 0.36 ng/mL, P < 0.05), but not in the pulmonary artery (0.10 +/- 0.03 v 0.56 +/- 0.21 ng/mL, P > 0.05). Blood PAF amounts in the radial artery were significantly higher than in the left atrium following ECC (1.09 +/- 0.36 v 0.06 +/- 0.04, P < 0.05), probably indicating PAF production in the heart. No variation of blood lipo PAF and lyso PAF was observed. No correlation was seen between PAF amounts and blood cell count.(ABSTRACT TRUNCATED AT 250 WORDS)


Should the gas outlet port on membrane oxygenators be routinely scavenged during cardiopulmonary bypass? Elimination of a volatile anesthetic agent administered prior to the start of bypass through the oxygenator has not been previously described. The purpose of this study was to determine the contamination risk from enflurane used before but not during cardiopulmonary bypass. Enflurane concentration was measured from the gas outlet port of a membrane oxygenator using infrared gas analysis in 11 cardiac surgical patients. The mean peak concentration at the gas outlet port in patients who had a final end-tidal concentration of < or = 0.3% was 0.08 +/- 0.04%, compared to 0.18 +/- 0.05% in patients with a final end-tidal concentration of more than 0.3%, P < 0.001. The elapsed time to reach peak concentration was 8.3 +/- 8.4 minutes, whereas the elapsed time from the peak concentration to 50% of the peak level was 63.1 +/- 25.1 minutes. At the time of peak enflurane release, a concentration of less than 2 ppm was measured at distances of 10 cm or less from the oxygenator gas outlet port in patients with an end-tidal enflurane of < or = 0.3%. In one patient with a final end-tidal enflurane of 1.1%, a contaminant level of 2 ppm could be measured at 95 cm from the oxygenator gas outlet port. This demonstrates that there is a potential risk of contamination from volatile anesthetics used immediately prior to extracorporeal circulation. Minimizing this risk may necessitate routine scavenging of the oxygenator, or simply avoiding increased concentrations of inhalation anesthesia before initiating cardiopulmonary bypass.


Isoflurane and hypothermic cardiopulmonary bypass: vasodilation without metabolic effects. During cardiopulmonary bypass, isoflurane may have beneficial effects on systemic oxygen uptake and vascular resistance. For this reason, the effects of isoflurane during low-flow (1.6 L/min/m2), hypothermic (27 degrees to 29 degrees C) cardiopulmonary bypass on systemic hemodynamics and oxygen uptake were studied in 20 patients in a cross-over experiment. Mean arterial and central venous pressures were measured during two consecutive periods of 10 minutes' duration. Blood samples were aspirated at the end of each period from the arterial and venous lines and analyzed for oxygen content. The concentration of isoflurane in the arterial samples was also determined. Systemic oxygen uptake and vascular resistance were calculated. Isoflurane had no significant effect on systemic oxygen uptake. Significant inverse relationships between blood isoflurane concentration and both mean arterial pressure and systemic vascular resistance were found. It is concluded that isoflurane is a vasodilator under the abnormal conditions of hypothermic cardiopulmonary bypass, but has no effect on systemic oxygen uptake.


The utility of a double-lumen tube for one-lung ventilation in a variety of noncardiac thoracic surgical procedures. To determine the utility of one-lung ventilation (OLV) in a variety of noncardiac thoracic surgical procedures, 200 patients were studied to document the ease of double-lumen tube (DLT) placement, associated complications, intraoperative respiratory changes, and methods for managing hypoxic events. Most tubes could be placed, repositioned when necessary, and secured within 12 minutes. By defining tube position with fiberoptic bronchoscopy, auscultatory assessment of placement was found to be incorrect in 38.0% of patients. The tip occluded the respective upper lobe orifice in 40.5% of this subgroup, the endobronchial cuff was at or above the carina in 38.7%, and in the wrong mainstem bronchus in 20.8%. During OLV, PaO2 initially fell to approximately 200 mmHg in most patients but gradually rose during the balance of the operation. Hypoxia (PaO2 less than 80 mmHg) during OLV developed in 28.5% of patients. Preoperative spirometry and arterial blood gases had no predictive value for this complication. Pulse oximetry values between 95% and 100% reliably reflected systemic arterial oxygen saturation. Hypoxia occurring during OLV was successfully reversed in 40.0% of instances by positive end-expiratory pressure (PEEP) to the ventilated lung. The addition of continuous positive airway pressure (CPAP) to the nonventilated lung reversed persistent hypoxia in virtually all cases. There was no difference in oxygenation, carbon dioxide elimination, airway pressures, or intraoperative complications noted between right and left double-lumen tubes. In conclusion, a DLT for OLV can expeditiously and safely be placed. Because auscultation for tube position is unreliable, bronchoscopic assessment of final position should be performed in every instance. Hypoxia during OLV can be detected reliably by pulse oximetry.(ABSTRACT TRUNCATED AT 250 WORDS)


Safety of a guidewire technique for replacement of pulmonary artery catheters. The purpose of this study was to determine if a guidewire change from a pulmonary artery catheter (PAC) to a central venous catheter (CVC) poses a significant infection risk. A total of 128 consecutive cardiac surgical patients with PACs inserted in the operating room were entered into this study. Postoperatively, patients were randomly allocated to receive a double-lumen CVC, either at the initial introducer insertion site over a guidewire, or at a new site with de novo catheterization. The tips of all introducers, PACs, and CVCs were cut off, cultured, and semi-quantitatively analyzed. The results show that insertion of CVCs over a guidewire within 48 hours after initial venapuncture is no more likely to be associated with catheter colonization than is de novo percutaneous insertion at a different site. From 48 hours up to 72 hours following initial insertion of the PAC, an incidence of catheter-related infection of 35.3% was observed in the guidewire group, as opposed to 12.5% in the de novo group. It is recommended that the use of a guidewire technique for catheter replacement (PAC to CVC) is a safe alternative to de novo insertion of a CVC within 48 hours after initial insertion of the PAC. In order to minimize the potential risk of catheter-related infection and bacteremia in cardiac surgical patients, de novo catheterization beyond 48 hours after initial venapuncture is suggested.


Con: whole blood transfusions are not useful in patients undergoing cardiac surgery. Data supporting fresh whole blood transfusion or fresh component therapy are nonblinded, and although both are conceptually attractive, neither can be considered proven. Recent blinded studies reflect fresh blood ineffectiveness. Larger, blinded, randomized trials will need to be performed. Proven methods of blood conservation as well as standardized criteria for transfusion of blood components will more effectively decrease homologous blood transfusion. Transfusion of fresh or banked whole blood, or its components, has yet to be shown to decrease the usage of homologous blood products.


[Fiber endoscopy in the extraction of foreign bodies from the gastrointestinal tract]. 23 patients that they swallowed foreign bodies, were treated. There were 16 male and 7 female, between 3 and 85 years old; 6 children and 17 adults. Of all, 7 were psychiatric patients. 15 foreign bodies were removed by upper digestive fiberendoscopy and 2, by procto/colonoscopy, without early nor late complications. 8 foreign bodies were eliminated spontaneously through the rectum/anus. Fiberendoscopy is an excellent method for treatment in these clinical problems.


[Surgical mortality and long-term recurrence of ileocecal volvulus]. Acute ileocecal volvulus is a condition of the abdomen which requires emergency treatment and is currently managed by several methods with a mortality rate of 25%. Surgical options include detorsion, cecostomy for nongangrenous volvulus or resection when gangrene is present. We studied the mortality and recurrence rate of ileocecal volvulus. This retrospective analysis evaluated 16 patients who underwent operation at Belen Hospital, Trujillo, Peru, between January 1966 and August 1992. As of August 1992, median follow-up was 160 months for 13 surviving patients (range, 6 to 307 months). Eleven men and 5 women with a median age of 52.3 + 21.1 years (range, 9 months to 83 years) formed the study population. Sixty eight percent of cases were 41 to 80 years of age and all women were than 40 years of age. Most of them were from the Peruvian Andes (81.3%), were from Indian and Spanish extraction (93.7%), and farmers (50%). Twelve patients (75%) developed a chronic ileocecal volvulus and 4 (25%) had an acute presentation. The clinical picture presented as large bowel obstruction in all patients. An ileocecal volvulus was accurately diagnosed by plain films of the abdomen in only 2 patients (16%) pre-operatively. A variety of procedures were used: cecopexy (9 cases), right hemicolectomy (3 cases) simple detorsion (2 cases), Mickulics exteriorization-resection (1 case) and cecostomy (1 case). The operative mortality rate was 18.7%. There were no recurrences after cecopexy. The most frequent post-operative complications were bronchopneumonia (18.7%), and wound infection (18.7%).(ABSTRACT TRUNCATED AT 250 WORDS)


[Malignant neoplasms of the gallbladder and extrahepatic bile ducts. A clinicopathological review of 81 cases]. We study the clinic, surgical pathologic aspects of 81 patients with cancer of the gallbladder and extrahepatic bile ducts, dividing them in 4 groups by localization. The majority of cases were women, the average in age was 57.05 years with the greatest incidence between the 5th and 6th decade of life. We can conclude that the symptoms and signs are not useful as well as the laboratory and radiologic test. Adenocarcinoma was the most common type of cancer and the outcome of the patients was very poor because cancer has grown beyond the limits of resectability before it is clinically apparent. Its important to be aware that 80% of the cancers of the gallbladder had lithiasis.


[Predisposing factors, clinical picture and mortality in volvulus of the small intestine]. This retrospective study evaluated predisposing factors, clinical picture and the methods of treatment related to morbidity and mortality of 19 small bowel volvulus (SBV) who underwent operation at Belen Hospital (Trujillo-Peru) during the last 26 years (1966-1992). The SBV was 1.6% of all cases of intestinal obstruction in this period and 10.8% of all intestinal volvulus. The median age was of 43 +/- 20.5 years (range, 6 to 78 years) and the majority of them were between 41 and 60 years. Sixteen cases (84.2%) were men from Indian and Spanish extraction and most of them were farmers and came from the Sierra of the Department of La Libertad. Two cases (10.5%) had non-related antecedents previous surgery. In six patients (31.6%) the volvulus was less than seven day's duration and in thirty (68.4%) it was more eight day's duration with previous attacks of obstruction (median: 19.3 days, range: 17 hours to 94 days). Pain, vomiting and distention were present in almost all of these cases. The most frequent abdominal finding was distention. The location of the volvulus was: ileum, 12 cases (63.2%), root of mesentery, 4 cases (21%) and jejunum, 3 cases (15.8%). Gangrenous bowel was present in six patients (31.5) and gangrenous intestine with perforation in two cases (10.5%) who underwent resection of the involved segment with primary anastomosis. In this group one patient (5.2%) died of sepsis and the wound infection rate was of 37.5%. There was no statistically significant correlation with the duration of illness and the presence of gangrenous loops or the mortality rate (p > 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)


Detection and prevention of colon cancer by colonoscopy. It is widely accepted that most carcinomas of the colon and rectum develop through the adenoma-to-carcinoma sequence. A recent review characterizes the various relationships between colorectal polyps and carcinoma. Tierney and Associates list several studies that show the frequent coexistence of adenomatous polyps with carcinoma of the colon or rectum. The incidence of polyps in patients with carcinoma of the colon and rectum ranges from 12.9% to 62%. Retrospective reports show a lower incidence of polyps in patients with carcinoma of the colon or rectum (13.7% to 23%). However, colon segments resected for carcinoma of the colon have a significant incidence of associated adenomatous polyps (15% to 28%). In patients with carcinoma of the colon or rectum undergoing preoperative fiberoptic colonoscopy there is a percentage of polyps larger than 5mm in diameter ranging between 36% and 62%. There is also a higher rate of both synchronous and metachronous carcinomas in patients in whom polyps coexist with the primary malignant lesion. The rate of metachronous carcinomas in patients with associated polyps at the time the first tumor was discovered is reported to be twice as great as for patients who did not have polyps (2.6% versus 1.14%, respectively). Synchronous malignant lesions were detected in 11% of patients with polyps but in only 0.7% of patients without polyps. In patients with multiple polyps coexistent with the original carcinoma, synchronous malignant lesions are found in 14.6% of patients and metachronous malignant lesions developed in 12.4%.(ABSTRACT TRUNCATED AT 250 WORDS)


Effect of triiodothyronine on rat liver polysome profiles and translational activity of mRNA after partial hepatectomy. The administration of 3,5,3'-L-triiodothyronine (T3) (200 micrograms/100 g) to rats immediately after partial hepatectomy led to an increase in total polysome content of residual hepatocytes (as measured 18 h postoperatively) which exceeded that observed after partial hepatectomy in the absence of hormone. The percentage of free ribosome pool remained the same after partial hepatectomy irrespectively on the absence or presence of T3. After partial hepatectomy the sedimentation pattern of total polysomal fraction (isolated 18 h postoperatively) shifted towards smaller aggregates as compared with that isolated from sham operated animals. T3 did not affect this pattern. The translational capacity of mRNA isolated from regenerating liver after 18 h was higher than that of mRNA isolated from sham operated animals. T3 increased the translational capacity of mRNA isolated from regenerating liver if administered immediately after surgery. Two dimensional electrophoresis of translated products showed that one of the typical proteins recently characterized with Mr 25,000 pI 6.9 appeared after partial hepatectomy and was significantly increased when T3 was administered immediately after surgery. These results show that thyroid hormone can enhance some of the characteristics features of liver regeneration involved in the synthesis of proteins typical for the process of liver regeneration.


N-methyl-D-aspartic acid injected peripherally stimulates oxytocin and vasopressin release. The effects of N-methyl-D-aspartic acid (NMDA), injected s.c. or i.p. in the dose range of 2.5-10 mg/kg, on oxytocin and vasopressin levels were assessed in conscious rats. NMDA administration was found to induce a dose-related increase in oxytocin concentration with a peak response at 7.5 min. Plasma vasopressin was elevated only after injection of the highest dose used (10 mg/kg). Thus, though at different thresholds, the release of both posterior pituitary hormones was stimulated after NMDA administration.


On the blood-brain barrier to peptides: effects of immobilization stress on regional blood supply and accumulation of labelled peptides in the rat brain. Tritiated arginine-vasopressin (AVP), desglycinamide-vasopressin (DGAVP), chicken gonadotropin releasing hormone (GnRH) or carbetocin were injected intracarotidally into rats exposed to a restraint stress for 60 min. The peptide accumulations were determined in 9-13 brain regions and anterior pituitary. In separate experiments the cerebral blood flow was measured. The blood supply to the brain was decreased in stressed animals as indicated by: 1. significant decrease (17-50%) of cerebral blood flow; 2. diminished accumulation of tritiated AVP in the regions lacking a blood-brain barrier (BBB). Consequently, the values of peptide accumulation were corrected for the changed blood supply. Compared with control animals, restraint stress induced a higher accumulation of AVP (+41%), DGAVP (+60%), carbetocin (+81%) and GnRH (+104%).


Neonatal erythropoiesis. I. Peripheral blood erythropoietic parameters: data suggest erythropoietin transfer via maternal milk. In this study peripheral blood erythropoietic parameters in 9 to 12-day-old neonatal rats suckled by experimentally-induced anemic mothers were examined. Stimulation of erythropoiesis in these pups was judged by increases in hematocrit, hemoglobin, mean corpuscular hemoglobin and mean corpuscular hemoglobin concentration. Failure to observe increases in reticulocytes may be, in part, the result of decreased maturation time, as indicated by reticulocyte Heilmeyer maturation indices in peripheral blood. The reticulocyte maturation curve was shifted to the left in neonates nursing from anemic mothers. These results suggest that erythropoietin (Ep) is transmitted to suckling rats via maternal milk, and by escaping inactivation in their gastrointestinal tract stimulates their erythropoiesis. These findings, in agreement with our previous work, are supportive of studies by others and further indicate a maternal-neonatal erythropoietic relation in rats.


Luteinization of ovaries and gonadotrophin and prolactin secretion in rats with posterior hypothalamic lesions. Posterior hypothalamic lesions restricted to the mammillary body in newborn rats evoked significantly elevated serum prolactin concentrations (P < 0.05) in adult females in the afternoon of proestrous (16.00 h), while at the same time serum LH values appeared significantly depressed (P < 0.05) as compared to controls. FSH concentrations were not affected. Parallel to changes in hormonal pattern, the ovaries of the lesioned animals grew to excessive dimensions due to the accumulation and persistence of numerous corpora lutea (CL) (syndrome of hyperluteinized ovaries). The results suggest that the posterior hypothalamus can regulate prolactin and LH secretion and that the fate of CL is associated with a quantitative ratio in the circulation of at least two hormones, prolactin and LH.


PCR detection of structurally abnormal Y chromosomes. Three probes each detecting a locus on the proximal long arm of the Y chromosome were partially sequenced. Thus, 3 sets of novel primers were developed which enable PCR detection of these 3 loci. Five previously reported primer sets, 3 on the short arm and each one on the centromere and the distal long arm, were mapped along with the novel three using a mapping panel consisted of 8 patients each with different structural abnormality of the Y chromosome. Now, PCR detection of these 8 loci covering an entire length of the Y chromosome has become possible enabling rapid screening of patients with Y chromosome aberrations.


Social identity and the International Classification of Handicaps: an evaluation of the consequences of facioscapulohumeral muscular dystrophy. The objective of this study was to evaluate the consequences of facioscapulohumeral muscular dystrophy (FSH MD) using two different but complementary procedures: an analysis based on the three different dimensions of disablement developed by the WHO--impairment, disability, and handicap--and a study of the psychological repercussions on social identity. Sixty-eight individuals with FSH MD, with 68 members of a control group, responded to a battery of psychosocial questions. Individuals with muscular dystrophy were also studied with reference to the dimensions of impairment, disability and handicap. The results showed that there are close correlations among measures of the three dimensions of disablement. Evaluations made by people with muscular dystrophy of the seriousness of their own disablement are strongly linked to objective measures of impairment. Furthermore, we found that having muscular dystrophy does have certain consequences for an individual's self-identity, although the degree to which one's self-image is validated is to some extent independent of the seriousness of the illness.


Rapid preparation of diagnostic probes for the fragile X syndrome by direct PCR amplification of human chromosomal DNA. The fragile X syndrome is a common familial form of mental retardation and is associated with a rare fragile site at Xq27.3 (FRAXA). This disorder has recently been reported to correlate with length variations of restriction genomic DNA fragments which may due to the amplification of (CCG)n trinucleotide repeats located at the FRAXA locus. We described here a rapid preparation method of diagnostic DNA probes for the fragile X syndrome by direct enzymatic amplification of human chromosomal DNA. The PstI-assay, which is Southern blot analysis of DNA samples probed by PCR products, was shown to be sensitive method for diagnostic purposes to detect the size variations specific in the fragile X syndrome.


Objective parameters in magnetic resonance imaging (MRI) for neuromuscular diagnosis: preliminary findings. T1 relaxation time (T1 time), T2 relaxation time (T2 time), and proton (rho) density in the thigh muscles of 20 normal healthy volunteers and three patients with muscle atrophy in the lower extremity were measured in order to select the useful MRI parameters for neuromuscular diagnosis. Since the standard deviation (SD) of both T1 and T2 times in each muscle was found to be within a relatively small range, these values were expected to be useful MRI parameters for neuromuscular diagnosis. On the contrary, rho density was not a valid parameter for diagnosis, as it was demonstrated to have large SDs in the muscles. The differences of these parameters in the three patients also supported the fact that MRI was useful for discriminating between the various types of muscular abnormalities based on relaxation times. The longer T1 and T2 times in women suggested that the content of water in skeletal muscle was higher in women than in men. Moreover, the T1 time in the dominant limbs was found to be shorter than in the non-dominant limbs for rectus femoris only in both men and women, while there were no differences in T2 time in these muscles.


A regional survey of the housing circumstances of families with children experiencing intellectual and motor disabilities. Housing is an important aspect in promoting the home care of disabled children, but also one which is too often forgotten by professionals. The purpose of this study was to evaluate the housing conditions and the need for modifications, of 204 families with disabled children using technical aids. The housing condition of the families was good compared to average Finnish families with children. One hundred and sixty families had made no housing modifications, but 48 families had need for them, most often to bathrooms and toilets. The need increased with the severity of the children's motor and intellectual disabilities and the need to provide care in line with the number of technical aids in use. It is concluded that families with disabled children need more information on the possibilities for carrying out housing modifications, and that home visits by a case manager are important to evaluate environmental factors affecting the care.


Researching disability: the way forward. This paper critically examines the individualistic and medicalized assumptions underlying much research on disability. The OPCS surveys of disability, in particular, are examined. Based on the perceptions of disabled people and their organizations, and drawing on a series of seminars on disability research which took place in 1991, an alternative social view of disability is presented and applied to research. It is concluded that the assumptions underlying much disability research, especially when they are translated into practice, are oppressive to disabled people, and that participatory and emancipatory research needs to be developed in order to assist disabled people in their struggle for empowerment.


Expressivity of a common fragile site, fra(3)(p14.2), in patients with cancer and other diseases. A population survey of a common folate-sensitive fragile site, fra(3)(p14.2), has been carried out on PHA-stimulated peripheral lymphocytes of patients with cancer and other diseases, under both culture conditions of folate deprivation and aphidicolin treatment. Overall findings regarding variability of expressivity due to age and sex were very similar to those obtained in a healthy population. The expression of fra(3)(p14.2) by folate deficient condition appeared hardly influenced by such exogenous factors as tobacco smoking habit, past histories of radiotherapy and chemotherapy, while it was associated with the unfavorable prognosis of cancers. Furthermore, proportion of those with higher expression was slightly but significantly larger in both lung and breast cancer patients. These findings suggest that some factors relevant to the expression of fra(3)(p14.2) may be associated with development and progression of certain kinds of cancer.


Chromosome abnormalities and rare fragile sites detected in azoospermia patients. We have examined constitutional chromosome abnormalities and fragile sites in 40 patients with azoospermia. Chromosome abnormalities were found in four cases. Three cases showed a deletion of the long arm of the Y chromosome 46,X,del(Yq) and the other case had a ring of G group chromosome 46,XY,r(G). In a rare fragile sites test, four fragile site carriers were detected and three rare autosomal fragile sites were identified; fra(8)(q24.1), fra(11)(p15.1), and fra(17)(p12). The expression of these fragile sites were induced specifically by AT-specific DNA ligands, such as distamycin A and Hoechst 33258. In addition, one patient was found to be the case of double ascertainment of fragile sites, fra(8)(q24.1) and fra(17)(p12). The overall frequency of distamycin A-inducible fragile sites in azoospermia patients appeared to be higher than those reported for Japanese healthy subjects and cancer patients. However, no significant relation among fragile sites, clinical and histological findings has been detected so far.


The control of genu recurvatum by combining the Swedish knee-cage and an ankle-foot brace. Genu recurvatum is a consequence of a poor control over the knee joint due to muscle weakness, impaired tonus and deficit in joint proprioception. Uncontrolled locking of the knee during ambulation causes recurrent microtrauma which leads to degenerative changes and instability. Known methods to control hyperextension of the knee during ambulation often fail. We have used the Swedish knee-cage attached to ankle-foot brace in an attempt to improve control over the knee in three patients. Gait analysis was performed to evaluate the effectiveness of this combined orthosis. Quality of gait improved, as demonstrated by a decrease in stance time asymmetry in the order of 45%; stride increased by 29% and speed of ambulation increased by 72%.


Physical parameters in Japanese newborns. A total of 19 physical parameters of the head, face, chest, and the fingers were examined in Japanese 50 male and 50 female newborns, measured 8 to 64 hr after birth. Exceptional values were excluded referring to the estimated mean and standard deviations. Normal values are presented as mean +/- 2 S.D. for each sex. As compared with Caucasian newborns, the Japanese newborns showed longer inner canthal and shorter outer canthal distances, shorter ear lengths, and longer palm and middle finger lengths.


Interstitial deletion of the long arm of chromosome 11: report of a case and review of the literature. A 12-month-old female infant with developmental delay, growth retardation, and dysmorphic features including dolichocephaly, telecanthus, ptosis, flat nasal bridge, anteverted nares, high-arched palate, carp-shaped mouth, micro-retrognathia, and low-set and posteriorly rotated ears was found to have an interstitial deletion of chromosome 11 involving bands q14-q22. Immunoblot analysis of her fibroblasts revealed a normal amount of mitochondrial acetoacetyl-coenzyme A thiolase, of which gene locus has been assigned to chromosome 11q22.3-q23.1. This result suggested that the region around the boundary of 11q22.3-q23.1 was intact in this patient.


A case of atypical Duchenne type muscular dystrophy with fragile X. The patient was a 9-year-old boy. He began to walk at the age 1 year and 8 months and began to speak at the age of 2 years, suggesting retarded mental and motor development. A diagnosis of DMD was made when he was 7 years old. On admission, the patient exhibited a peculiar thin and long face, large auricles, narrow palate, malalignment of the teeth, epicanthus, saddle nose, and simian lines in addition to symptoms consistent with DMD. Chromosome analysis showed fragile X at Xq27 at a frequency of 20%. His mother also showed fragile X at the same position. Since the atypical features of this DMD patient are all explained as fragile X syndrome, this case was considered to be a very rare instance of DMD whose clinical pictures were modified by fragile X syndrome.


Neurospora crassa blue-light-inducible gene bli-7 encodes a short hydrophobic protein. Blue light induces a number of physiological reactions in Neurospora crassa. We have cloned and sequenced the gene bli-7, which is inducible by blue light, and both glucose and nitrogen starvation. This gene is strongly expressed at the RNA-level and contributes up to 0.2% of N. crassa total RNA when fully induced. The deduced amino acid sequence reveals a short (108 amino acids), hydrophobic protein which has homology to the protein SC-3, encoded by a gene of Schizophyllum commune which was isolated as a gene abundantly expressed at the time of fruiting. The amino-terminus of BLI-7 protein shows resemblance to a number of transit peptides. The comparison of the nucleotide sequence of the promoter region with various N. crassa promoters reveals a striking similarity to the promoter of grg-1, a glucose repressible gene.


The replication origin of the Methylomonas clara plasmid pBE-2. The Methylomonas clara narrow host range plasmids pBE-2 and pBE-3 belong to the class of plasmids encoding a trans acting replication initiation factor. Characteristically for such plasmids, the sequence of the origin of pBE-2 and pBE-3 contains a number of large direct repeats (8 and a half iterons of 19 bp), which by analogy are putative binding sites of the trans acting replication factor. Several additional features typical for the majority of E. coli plasmids were found in the M. clara origin: These include sequences homologous to the E. coli DnaA-box, sequences resembling E. coli IHF binding-sites, an AT-rich region with short repeats (similar to those repeats of E. coli origins responsible for an initial DNA duplex opening), and an AT-rich bent DNA region containing inverted repeats which have homology to small repeated sequences found in several plasmid origins. In addition, in the M. clara plasmid origin, large potential hairpin structures are present and the sequence of one of these participates in site specific recombination.


Molecular cloning of mouse alcohol dehydrogenase-B2 cDNA: nucleotide sequences of the class III ADH genes evolve slowly even for silent substitutions. We have cloned and sequenced a cDNA encoding the mouse class III alcohol dehydrogenase, Adh-B2. Adh-B2 mRNA is detectable in all the mouse tissues tested. Class III ADHs are highly conserved: the deduced amino acid sequence of the mouse Adh-B2 is 91 to 97% identical to the human, horse and rat liver enzymes. The mouse Adh-B2 cDNA is 87% identical in nucleotide sequence to the human chi-ADH cDNA. Previously, a slower rate of evolutionary divergence of the amino acid sequences of class III ADH proteins was detected and ascribed to functional constraints upon the protein. Our analysis of the nucleotide sequences demonstrates that this cannot be the entire explanation, since the rate of silent (synonymous) nucleotide substitutions is also lower in the class III ADHs than in the class I ADHs.


The HIT protein family: a new family of proteins present in prokaryotes, yeast and mammals. By comparing the sequence of a putative translation product from a Saccharomyces cerevisiae split gene with several data-bases, I have uncovered a new protein family. Members of this family are found in prokaryotes as well as in lower and higher eukaryotes. The function of these proteins is unknown but they share a characteristic histidine triad that may be involved in zinc binding. This group of protein has been named the HIT protein family.


Cloning and sequencing of European sea bass (Dicentrarchus labrax L.) growth hormone cDNA using polymerase chain reaction and degenerate oligonucleotides. To clone and sequence the cDNA of the growth hormone of European sea bass (Dicentrarchus labrax L.) (sbGH), total pituitary RNA was reverse transcribed and amplified using polymerase chain reaction (PCR). Degenerate oligonucleotides, designed by comparing available GH cDNA sequences from related teleost species, were used as primers to amplify the 5' end and the core region of sbGH cDNA, while the 3' end was amplified according to the Rapid Amplification of cDNA ends (RACE) method. SbGH cDNA contains an open reading frame encoding a preprotein of 204 amino acids. The deduced amino acid sequence shows a putative signal peptide of 17 amino acids, suggesting that the mature hormone consists of 187 amino acids. Sequence comparison indicates a high degree of conservation of GH cDNAs within the Percoidei infraorder. Our procedure based on degenerate oligonucleotides and PCR provides a straightforward approach to clone GH cDNAs from other related bony fishes.


The nucleotide sequence of a putative membrane transport gene from Clostridium perfringens. A gene from Clostridium perfringens encoding a highly hydrophobic polypeptide of M(r) = 30,698 was cloned and sequenced. The polypeptide showed similarities with the inner membrane proteins of binding protein-dependent transport systems of the gram negative enterobacteria. The sequence homology to the UgpE and MalG proteins of Escherichia coli was 27.6%, or 60.4 and 52.2%, respectively, taking into account the conservative amino acid changes. The polypeptide contained the EAA---G---------(I/V)-LP motif of the gram negative transport proteins. This motif may thus be very old, as clostridia and E. coli separated some 1.5 x 10(9) years ago. The similarities suggest a common evolutionary origin of these genes.


The primary structure of Providencia rettgeri penicillin G amidase gene and its relationship to other gram negative amidases. The nucleotide sequence of Penicillin G amidase (PA,E.C.3.5.1.11) of Providencia rettgeri was determined. We aligned our P. rettgeri PA with other known Gram negative periplasmically located beta-lactam amidases. The analysis revealed a high homology with other Enterobacteric amidases (60%-65%), while with similar Pseudomonas sp. amidases the homology exceeded 25%. These homologies indicate their common ancestry.


Disease development in ethylene-insensitive Arabidopsis thaliana infected with virulent and avirulent Pseudomonas and Xanthomonas pathogens. The plant hormone ethylene has been hypothesized to play roles both in disease resistance and in disease susceptibility. These processes were examined by using isogenic virulent and avirulent bacterial pathogens and mutants of Arabidopsis thaliana that were altered in ethylene physiology. Ethylene-insensitive ein1 and ein2 mutants of Arabidopsis were resistant to Pseudomonas syringae pv. tomato made avirulent by the addition of the cloned avirulence genes avrRpt2, avrRpm1, or avrB; this suggests that ethylene is not required for active resistance against avirulent bacteria. In a second set of experiments, susceptibility was monitored with virulent P. s. pv. tomato, P. s. pv. maculicola, or Xanthomonas campestris pv. campestris strains. Wild-type Arabidopsis and ein1 mutants were susceptible to these strains, but ein2 mutants developed only minimal disease symptoms. Despite these reduced symptoms, virulent P. s. pv. tomato grew extensively within ein2 leaves. The Pseudomonas phytotoxin coronatine induces ethylene biosynthesis and diseaselike symptoms on many plant species, but the reduced symptomology of ein2 mutants could not be attributed to insensitivity to coronatine. The enhanced disease tolerance of ein2 plants suggests that ethylene may mediate pathogen-induced damage, but the absence of tolerance in ein1 mutants has yet to be explained.


Replication and movement of a coat protein mutant of cymbidium ringspot tombusvirus. The spread of cymbidium ringspot tombusvirus (CyRSV) in host tissue was studied by using a coat protein gene mutant with a six-nucleotide deletion; the deletion removes two amino acids from the shell domain (S) of the capsid protein. Mutated protein subunits were synthesized in infected cells but could not assemble into virus particles. Virions were formed, however, with inoculation of mutated RNA in transgenic plants expressing normal CyRSV coat protein. The mutant is restricted in long-distance movement in Nicotiana clevelandii, whereas it spreads systemically in N. benthamiana. These results suggest that tombusviruses may spread either as complete virions or in a nonvirion form, depending on the host plant species.


Determinants of pathogenicity in Xanthomonas campestris pv. vesicatoria are related to proteins involved in secretion in bacterial pathogens of animals. One of the model systems investigated for studying plant bacterial pathogenesis is Xanthomonas campestris pv vesicatoria, the causal agent of bacterial spot disease of pepper and tomato. Genes necessary for both basic pathogenicity and the induction of the hypersensitive response in resistant plants (hrp genes) were previously isolated from X. c. pv. vesicatoria and characterized genetically. As a first step toward functional analysis, part of the hrp gene cluster, making up several loci, was sequenced. Here, we report the first indications of the function of hrp genes. Striking similarities to proteins from the mammalian pathogens Shigella flexneri, Yersinia enterocolitica, Y. pestis, and other bacteria were discovered. Proteins encoded by genes within the X. c. pv. vesicatoria loci hrpA, hrpB, and hrpC are similar to ATPases and to Yersinia Ysc and LcrD proteins, which are involved in secretion of Yop proteins, a particular class of essential pathogenicity factors produced by Yersinia species. This finding indicates, for the first time, that the fundamental determinants of pathogenicity may be conserved among bacterial pathogens of plants and animals. We hypothesize that hrp genes are involved in the secretion of molecules essential for the interaction of X. c. pv. vesicatoria with the plant.


hrp genes of Pseudomonas solanacearum are homologous to pathogenicity determinants of animal pathogenic bacteria and are conserved among plant pathogenic bacteria. The majority of bacterial plant diseases are caused by members of three bacterial genera, Pseudomonas, Xanthomonas, and Erwinia. The identification and characterization of mutants that have lost the abilities to provoke disease symptoms on a compatible host and to induce a defensive hypersensitive reaction (HR) on an incompatible host have led to the discovery of clusters of hrp genes (hypersensitive reaction and pathogenicity) in phytopathogenic bacteria from each of these genera. Here, we report that predicted protein sequences of three hrp genes from Pseudomonas solanacearum show remarkable sequence similarity to key virulence determinants of animal pathogenic bacteria of the genus Yersinia. We also demonstrate DNA homologies between P. solanacearum hrp genes and hrp gene clusters of P. syringae pv. phaseolicola, Xanthomonas campestris pv. campestris, and Erwinia amylovora. By comparing the role of the Yersinia determinants in the control of the extracellular production of proteins required for pathogenicity, we propose that hrp genes code for an export system that might be conserved among many diverse bacterial pathogens of plants and animals but that is distinct from the general export pathway.


Epidemiology of bovine anaplasmosis and babesiosis in Latin America and the Caribbean. The various parameters which interact in the epizootiology of babesiosis and anaplasmosis and which it is necessary to obtain in a survey are analysed and the two diseases compared. Where no data are available from Latin America and the Caribbean, the references from Australia and the United States of America are discussed. The two principal objectives of a study on the epidemiology of babesiosis and anaplasmosis are: to determine the risk of occurrence to understand the relevant factors leading to outbreaks. The first objective is relatively simple to achieve for both diseases by calculating the animal inoculation rate (h) at a determined age and the critical value of h for enzootic stability. The second objective requires exhaustive and complex studies. For babesiosis epidemiology, studies of the tick infection rate, cattle infestation rate, genetic composition of cattle, acaricides, stocking rate, climatic data and other factors are necessary. Anaplasmosis epidemiology cannot be satisfactorily explained by considering it purely as a tick-borne disease. Difficulties to be overcome include the great diversity of haematophagous Diptera in Latin America, and the scarcity of data on the capacity of these insects as anaplasmosis vectors and their feed preferences on cattle.


Identification of a new inducible nodulation gene in Azorhizobium caulinodans. The narrow host range bacterial strain Azorhizobium caulinodans ORS571 induces the formation of nitrogen-fixing nodules on the root and stem of the tropical legume Sesbania rostrata. Here, a new flavonoid-inducible locus of ORS571 is described, locus 4. The locus was identified and isolated via the occurrence of particular sequences, the gamma and delta elements. These elements are reiterated in the ORS571 genome, linked to symbiotic loci. Sequencing of locus 4 showed the presence of an open reading frame (ORF6) that is flanked downstream by a gamma element and upstream by a delta element. The gamma element is approximately 180 bp in size, and shows homology to the insertion element ISRm3, an insertion sequence belonging to a distinct class of IS elements. The delta element is about 300 bp in size and has homology with repeated sequences found in other Rhizobiaceae. The ORF6 gene product shows a low, but significant homology to the mouse mastocytoma antigen P35B (Szikora et al., EMBO J. 9: 1041-1050, 1990) and to a class of NAD/NADP-binding sugar epimerase/dehydrogenases (Pissowotzki et al., Mol. Gen. Genet. 231: 113-213, 1991). Immediately upstream from ORF6, a nod box-related sequence is present, the arrangement of which is fully consistent with a recently presented model for the nod box structure (Goethals et al., Proc. Natl. Acad. Sci. USA 89: 1646-1650, 1992). Insertional inactivation of ORF6 did not affect the nodulation and fixation performance on S. rostrata. However, on S. formosa roots the nodulation kinetics of such a mutant was clearly affected (about 5 days delay). We propose to call this new symbiotic gene nolK.


Sequential expression of two late nodulin genes in the infected cells of alfalfa root nodules. The Nms-22 and leghemoglobin (Lb) genes are expressed exclusively in the infected cells of alfalfa root nodules. Expression of these two late nodulin genes originated at distinct cellular boundaries within the symbiotic region of the nodule. The Nms-22 gene was expressed in all infected cells, including those just adjacent to the meristematic region. Lb gene expression was induced in older infected cells and was most prominent in the mature region of the nodule. Despite this temporal separation of gene expression, both the Nms-22 and Lb genes were expressed in nodules elicited by bacA mutants in which bacteroid development has been blocked just after release from the infection thread.


Practical significance of rabies antibodies in cats and dogs. Doubt has sometimes been cast upon the protective effect of rabies antibodies in serum. Animals and humans suffering from fatal rabies often produce high antibody titres, while rabies cases are also observed in vaccinated animals. Cellular immunity is also largely involved in protection. Nevertheless, a large number of laboratory experiments and field observations clearly demonstrate that cats and dogs which develop antibodies after vaccination and before challenge have a very high probability of surviving any challenge, no matter how strong the dose and which virus strain was used. Rabies antibody titration can, therefore, afford a strong additional guarantee to the vaccination certificates accompanying domestic carnivores during transportation between countries. Quarantine rules should also be adapted to the epidemiological features in the exporting country, e.g. statistics of vaccination failure in cats and dogs and host-virus adaptation of the rabies strains circulating in these countries.


Observations on rinderpest in Kenya, 1986-1989. Rinderpest was confirmed in Kenya in 1986, 1987, 1988 and 1989. Three epidemiologically distinct events appear to have occurred: repeated outbreaks in West Pokot district related to cross-border movement of stock, an outbreak in Marsabit district in 1987 (thought to have been caused by illegal movement of cattle, possibly in vehicles, from countries further north) and a series of related outbreaks in and near Nairobi between 1988 and 1989 due to the unauthorized movement from abattoirs and holding grounds of slaughter stock possibly introduced from West Pokot or Marsabit. In West Pokot the disease affected unvaccinated calves and yearlings. In Marsabit cattle of all ages were affected. In August 1988, a major outbreak was confirmed in Kiambu and Kajiado districts in central Kenya, near Nairobi. At the same time a provisional diagnosis of rinderpest was made in a herd of cattle at a slaughterhouse in Nairobi. Rinderpest virus was isolated from sick cattle in all the outbreaks. Experimental infection of susceptible cattle with the Kiambu isolate demonstrated this to be of low virulence. Emergency vaccination and quarantine measures instituted immediately after confirmation eliminated clinical disease within three to four weeks in West Pokot, Kiambu and Nairobi. In Kajiado, however, the disease persisted for at least nine months, during which time a series of virus isolates was recovered. There was no evidence of infection in susceptible wildlife. This increase in the incidence of rinderpest in Kenya in recent years serves to highlight the problems of control and the need for concerted efforts to eradicate the threat of the disease from East Africa.


Foot and mouth disease in gayals (Bos gaurus frontalis) in Calcutta Zoo. A brief description is given of an outbreak of foot and mouth disease in gayals (Bos gaurus frontalis) which occurred in Calcutta Zoo in October 1990. Four of the five affected animals died within ten days of onset of the disease. The disease did not spread to other animals in the Zoo.


Homology of Rhizobium meliloti NodC to polysaccharide polymerizing enzymes. Rhizobium bacteria form nitrogen-fixing nodules on legume roots. As part of the nodulation process, they secrete Nod factors that are beta-1,4-linked oligomers of N-acetylglucosamine. These factors depend on nodulation (nod) genes, but most aspects of factor synthesis are not yet known. We show here that one gene, nodC, shows striking similarity to genes encoding proteins known to be involved in polysaccharide synthesis in yeast and bacteria, specifically chitin and cellulose synthases, as well as a protein with unknown function in Xenopus embryos, DG42. This similarity is consistent with a role for the NodC protein in the formation of the beta-1,4-linkage in Nod factors.


The Rhizobium, Bradyrhizobium, and Azorhizobium NodC proteins are homologous to yeast chitin synthases. The nodABC genes of rhizobia are essential for the synthesis of lipo-oligosaccharidic (N-acylated chitin oligomers) nodulation signals. nodC gene products from Rhizobium, Bradyrhizobium, and Azorhizobium exhibit extensive homology with chitin synthases, suggesting that the NodC proteins are involved in the synthesis of the chitin oligomer backbone by catalyzing the beta-1,4-linkage between N-acetyl-D-glucosamine residues.


Resurgence of contagious bovine pleuropneumonia in Nigeria. Epidemiological reports and post-mortem examination of slaughtered cattle indicate that prevalence of contagious bovine pleuropneumonia (CBPP) in Nigeria is on the increase, despite annual mass vaccinations and other protection measures. The number of reported outbreaks increased from 20 in 1981 to 64 in 1988, 114 in 1989, 82 in 1990 and 52 in 1991 (January to June). Post-mortem examinations conducted between January 1988 and June 1991 at the abattoir in Maiduguri, capital of Borno State (Nigeria) revealed that of 122,567 slaughtered cattle, 6,008 (4.9%) had pneumonia and 777 (0.6%) had gross lesions suggestive of CBPP. Only 22 sporadic outbreaks were reported over this period within the catchment area of Borno State. Probable reasons for the deteriorating situation of CBPP in Nigeria are discussed.


[Prevalence of Newcastle disease in traditional breeding facilities for chickens in Cameroon]. Although 65% of domesticated birds in the Cameroon are kept in traditional chicken farms, very little attention is given to this important sector by the administration as well as by the farmers themselves. This could be detrimental to the modern poultry sector where a lot of investments have been made during recent years. In order to estimate the danger of neglecting traditionally-raised chickens and the health threat they represent for the poultry population of the country, a study was conducted to evaluate the impact of Newcastle disease in traditional farms. A total of 180 chicken blood serum samples from three provinces of the Cameroon were submitted to the haemagglutination inhibition test; 51.66%, 48.33% and 46.66% of birds respectively from the East, West and North provinces reacted positively to the test. The consequences of such a situation are emphasised.


Detection of haemagglutination inhibition antibodies against Newcastle disease virus in unvaccinated indigenous chickens in Maiduguri, Borno State, Nigeria. An examination of 200 serum samples from unvaccinated indigenous (local) chickens in Maiduguri, Borno State (Nigeria) using the haemagglutination inhibition (HI) test showed 73 sera to be positive and 127 to be negative for antibodies against Newcastle disease virus. The highest antibody titre observed was 1:128. The prevalence rate was higher (46.9%) in adult chickens than in young chickens of less than 12 weeks (23%). Presence of HI antibodies in unvaccinated indigenous chickens indicates that these birds had contracted infection and recovered thereafter.


[Experimental infection of sheep with a rabies virus of canine origin: study of the pathogenicity for that species]. The effects of the inoculation of a canine strain of rabies virus in sheep were studied using ten animals which received different amounts of this virus. Two subjects, inoculated with 10(5.4) mouse intracerebral lethal doses 50% (MICLD50), died from rabies after 19 and 40 days of incubation. Clinical signs were anorexia, emaciation, nervous reactions and prostration before death. The virus was recovered from different parts of the central nervous system and salivary glands with high titres. Only three animals showed an antibody response, at very low levels.


Serological and bacteriological study of brucellosis in camels in central Saudi Arabia. Sera from 2,630 apparently normal adult camels (Camelus dromedarius) raised in central Saudi Arabia (Riyadh and Al-Kharj cities) were examined serologically by the Rose Bengal and standard United States of America Brucella plate agglutination tests. The overall seroprevalence of brucellosis in the restricted populations of tested camels was 8%. The seroprevalence of brucellosis among camels raised in small numbers in the backyards of 24 houses in Riyadh and those intensively raised on one large camel farm near Al-Kharj were 4.3% and 8.6% respectively. Fresh milk samples from 100 brucellosis seropositive camels from Riyadh and Al-Kharj were cultured on Brucella-selective media. Brucella melitensis biovars 1 and 2 were isolated and identified from 26 camels. Epidemiologically, brucellosis in camels in central Saudi Arabia appeared to be connected with B. melitensis infection of sheep and goats, and also represents a serious public health risk.


Treatment of Brucella melitensis infection in sheep and goats with oxytetracycline combined with streptomycin. Six treatment regimens using oxytetracycline (OTC) combined with streptomycin (ST) were evaluated for eliminating Brucella melitensis from 480 naturally-infected sheep and goats. Cessation of shedding Brucella from udder secretions and absence of Brucella in selected tissues at autopsy were considered criteria for successful treatment. Four regimens were equally effective in eliminating Brucella in the treated groups of sheep and goats regardless of the source of antibiotics used. These were regimen A (OTC 20 mg/kg intravenously daily for 6 weeks, combined with ST 20 mg/kg intramuscularly [i.m.] daily for 3 weeks), regimen B (long-acting [LA]-OTC 20 mg/kg i.m. every 3 days for 6 weeks, with ST 20 mg/kg i.m. every 3 days for 3 weeks), regimens D and E (LA-OTC 28 mg/kg i.m. every 3 days for 6 weeks, with ST 20 mg/kg i.m. every 3 days for 3 weeks). However, regimen C (LA-OTC 20 mg/kg i.m. every 3 days for 6 weeks, with ST 20 mg/kg i.m. every 3 days for 3 weeks) eliminated Brucella in only 75 of 80 (94%) sheep and goats. Regimen F (LA-OTC 25 mg/kg i.m. every 2 days for 4 weeks, combined with ST 20 mg/kg i.m. every 2 days for 2 weeks) was the most practical, effective and least expensive regimen for eliminating Brucella in the 80 treated sheep and goats. Brucella melitensis biovar 2 was repeatedly isolated from the mammary secretions of all sheep and goats before treatment. It was also isolated repeatedly from the udder secretions of all non-treated control animals and from selected tissue specimens collected from the controls at necropsy.


[Diagnosis of caprine contagious pleuropneumonia: recent improvements]. Due to the difficulty in isolating Mycoplasma sp. type F38, two techniques for rapid detection are proposed in order to identify the presence of mycoplasmas of the mycoides group in samples of pleural fluid. A modification in the composition of isolation media promotes the growth of type F38 mycoplasmas and inhibits the growth of M. ovipneumoniae strains.


Bovine virus diarrhoea-mucosal disease and border disease in Egypt. In Egypt, bovine virus diarrhoea-mucosal disease (BVD-MD) was initially detected as the result of a serological survey of cattle and sheep, using the serum neutralisation test. In 1970, the causal pestivirus was isolated from bovine calves and buffalo calves with pneumonia and enteritis. Mixed viral infections were also prevalent. Cases of immune tolerance were identified. The Egyptian industry had complained of unthrifty cattle and high death rates of bovine calves and buffalo calves. A rinderpest outbreak in 1982 involved some cattle and buffalo which had been vaccinated against the disease, and such animals were positive to the gel diffusion test for BVD-MD pestivirus. Extensive immunosuppression due to BVD MD virus was suspected, because the attenuated cell-culture vaccine against rinderpest, issued in Egypt since 1965, had not been tested for freedom from non-cytopathic BVD-MD virus. Experimental infection of susceptible sheep with BVD-MD virus resulted in symptoms of Border disease. Innocuity of the attenuated C24V bovine pestivirus vaccine for animal tissues and the immune system of calves was confirmed.


Pestivirus infections in ruminants in Norway. Serological surveys in Norway have demonstrated neutralising antibodies against bovine virus diarrhoea (BVD) virus in cattle, sheep and goats. The prevalences were 18.5%, 4.5% and 3.6%, respectively. Occurrence of pestivirus-induced disease in Norway is described. Outbreaks of reproductive failure and mucosal disease have been reported, and the number of persistently-infected animals detected has increased considerably in recent years. Acute BVD occurs rarely. Border disease (BD) in sheep, first diagnosed in 1981, has subsequently been demonstrated sporadically. In goats, typical BD was diagnosed in 1982, with three later occurrences of reproductive failure. Experimental infections in pregnant goats induced a high rate of severe foetopathogenic effect. Signs and lesions in offspring were comparable to ovine BD. Similar findings were demonstrated in goats given a pestivirus-contaminated vaccine. In newborn kids, experimental infection had an adverse influence on growth and health. Persistent infection in goats is probably rare.


Tissue concentrations of a long-acting oxytetracycline formulation after intramuscular administration in cattle. The tissue distribution of a long-acting oxytetracycline formulation after intramuscular administration to calves at a dose rate of 20 mg/kg was studied. Oxytetracycline concentrations were determined in lung, bone marrow, mammary gland, uterus, uterine horn, ovary, liver, synovial fluid, joint tissue, kidney, spleen, brain, muscle, fat, urine, bile, saliva, ruminal content and serum. Observed concentrations were higher than the minimal inhibitory concentration for the majority of pathogens in all of the analysed tissues for at least 72 h post-injection. The shortest theoretical permanency time was 1.88 days (in saliva) and the longest was 19.06 days (in kidney). Taking the maximal permitted residue level as 0.1 microgram/g in muscle, 0.3 microgram/g in liver and 0.6 microgram/g in kidney, the calculated withdrawal time was 20 days.