Exploring the Coordination of Plutonium and Mixed Plutonyl-Uranyl Complexes of Imidodiphosphinates.

The coordination chemistry of plutonium(IV) and plutonium(VI) with the complexing agents tetraphenyl and tetra-isopropyl imidodiphosphinate (TPIP- and TIPIP-) is reported. Treatment of sodium tetraphenylimidodiphosphinate (NaTPIP) and its related counterpart with peripheral isopropyl groups (NaTIPIP) with [NBu4]2[PuIV(NO3)6] yields the respective PuIV complexes [Pu(TPIP)3(NO3)] and [Pu(TIPIP)2(NO3)2] + [PuIV(TIPIP)3(NO3)]. Similarly, the reactions of NaTPIP and NaTIPIP with a Pu(VI) nitrate solution lead to the formation of [PuO2(HTIPIP)2(H2O)][NO3]2, which incorporates a protonated bidentate TIPIP- ligand, and [PuO2(TPIP)(HTPIP)(NO3)], where the protonated HTPIP ligand is bound in a monodentate fashion. Finally, a mixed U(VI)/Pu(VI) compound, [(UO2/PuO2)(TPIP)(HTPIP)(NO3)], is reported. All these actinyl complexes remain in the +VI oxidation state in solution over several weeks. The resultant complexes have been characterized using a combination of X-ray structural studies, NMR, optical, vibrational spectroscopies, and electrospray ionization mass spectrometry. The influence of the R-group (R = phenyl or iPr) on the nature of the complex is discussed with the help of DFT studies.

Safety behaviours among Alaskan Native and American Indian people living in Alaska.

OBJECTIVE: To examine self-reported safety behaviours among 3828 Alaskan Native and American Indian people enrolled in the Alaska Education and Research Towards Health (EARTH) Study, 2004-2006. DESIGN: A cross-sectional analysis of baseline data from a cohort study. A non-random sample of participants (2322 women and 1506 men) aged > or =18 years from three regions of Alaska completed questions on safety behaviours as part of a comprehensive health and lifestyle computer-assisted self-administered questionnaire. RESULTS: Most participants reported never driving after drinking (94.1%) or riding with a driver who had been drinking (91.3%). Fewer (74.1%) participants reported using a seatbelt always or almost always when riding in a vehicle. Only about half (55%) always kept to the speed limit when driving or used a personal flotation device when boating (51.5%). Even fewer (20.5%) reported using a helmet when riding on off-road vehicles, including four-wheelers and snowmobiles. Factors identified among those least likely to use safety devices and practise good safety behaviours are: younger age, lower household income and education, non-married, speaking only English at home, and a self-reported health status of poor to fair (p<0.05). CONCLUSIONS: Recommendations for future injury prevention efforts in this population are to increase use of personal flotation devices while boating and address the underuse of helmets with off-road vehicles. Limited prevention resources should be used to target those who engage in risky behaviours to maximise programme impact.

Cadaveric allograft microbiology.

This study aims to determine the contamination rate of cadaveric bone allograft and blood cultures retrieved from 119 donors within Leicester between 1990 and 2003. A contamination rate of 27% was present, with 120 of 437 bone allografts culturing positive at the time of retrieval. Similarly, a contamination rate of 37% was present, with 40 of 107 blood samples culturing positive. The time interval between death and procurement did not influence blood contamination. Coagulase-negative Staphylococcus was the commonest organism isolated in both blood and bone cultures. One donor had Clostridium grown in their blood culture. The available evidence confirms similar contamination rates with other studies. The majority of organisms isolated were skin commensals with a low rate of contamination of highly pathogenic organisms such as Clostridium.

A murine monoclonal anti-idiotype to anti-ribosomal P antibodies: production, characterization, and use in systemic lupus erythematosus.

Overt anti-ribosomal P (anti-P) autoantibodies are restricted to a subset of systemic lupus erythematosus (SLE) patients, and are potentially pathogenic. Covert anti-P are detected in all other individuals. An idiotype (Id) network is nonoperational in those with overt anti-P, whereas it is functional in all others. The aim of this study was to produce a murine monoclonal (mAb) anti-Id to characterize the anti-P Id network in SLE. BALB/c mice were immunized with F(ab')(2) fragments of IgG anti-P from a patient with a broadly cross-reactive Id. One mAb was chosen (mAb41) that reacted preferentially to the immunogen. This IgG1 mAb bound comparably in ELISAs to affinity-purified anti-P from 11 SLE patients with overt anti-P. This binding was partially inhibited with ribosomal P antigen. In contrast, it did not bind to affinity-purified control autoantibodies, nor to normal human IgG. mAb41 inhibited anti-P binding to ribosomal P antigen in immunoassays and on Jurkat cells. No change was detected in patients' anti-P antibodies over time when mAb41 was used in Id-specific ELISAs. We conclude that mAb41 is an anti-Id that recognizes a public idiotope within the antigen-combining site of anti-P antibodies. Thus, it is analogous to its human counterparts, and potentially, would modulate the pathogenicity of anti-P autoantibodies in vivo.

Autoantibodies to the 20-kDa ribosomal proteins: identification, characterization, and new aspects on prevalence in systemic Lupus erythematosus.

Autoantibodies to the 20-kDa ribosomal proteins (L12/S10) are not well studied, especially in juveniles with systemic lupus erythematosus (SLE). Randomly selected sera from American juveniles and adults with SLE were screened for antibodies to either 20-kDa protein and P proteins and then assayed for anti-L12 and anti-S10 by immunoblot assays. In a pilot study of patients with anti-P (Cohort 1), IgG antibodies to either 20-kDa protein and, specifically, to L12 were observed in 72 and 42% of juveniles and adults, respectively. IgG antibodies to S10 were detected less frequently. In Cohort 2 patients who were chosen irrespective of autoantibody status, twice as many juveniles as adults had IgG antibodies to either 20-kDa protein. Prevalences of IgG anti-L12 and IgG anti-S10 antibodies in the juveniles were 28 and 16% and in the adults were 13 and 12%, respectively. Anti-L12 were strongly but not invariably associated with anti-P, and usually arose temporally to these antibodies. Anti-S10 activity was due to anti-Sm antibodies. We conclude that IgG anti-L12 are more prevalent in SLE than previously reported, and are responsible for the majority of activity toward the 20-kDa ribosomal proteins, especially in juveniles.

Aortic valve replacement for endocarditis: determinants of early and late outcome.

BACKGROUND AND AIM OF THE STUDY: The study aim was to determine risk factors for operative mortality, recurrent infection, reoperation and long-term survival following aortic valve replacement (AVR) for infective endocarditis. METHODS: Between 1973 and 1997, 109 patients (91 male, 18 female, mean age 52.6 years) underwent isolated AVR for infective endocarditis in our unit. Native valve endocarditis was present in 89 (81.6%) and prosthetic valve endocarditis in 20 (18.4%). Active culture-positive endocarditis was present in 53 (48.6%). Preoperatively, 99 patients (90.8%) were in NYHA classes III and IV. Indications for surgery included cardiac failure in 41 patients, valvular dysfunction in 38, vegetations in 18, sepsis in seven, abscess in six and embolism in four. Mechanical valves were implanted in 69 patients (63.3%) and bioprostheses in 40 (36.7%), including a homograft in 19 (17.4%). Follow up was complete (mean 5.8 years; range: 0-23.8 years; total 633.5 patient-years). RESULTS: The operative mortality was 10.1% (11 deaths). At ten years, freedom from recurrent infection was 94.2%, and freedom from reoperation 83.6%. Biological valve and younger age were significant adverse parameters for freedom from reoperation (p = 0.01 and p = 0.01). There have been 21 late deaths, 15 due to cardiac causes. Kaplan-Meier survival, including operative mortality, at five and ten years was 77.4% and 68.0%, respectively. On Cox proportional hazards regression, Staphylococcus aureus infection (p = 0.008) and older age (p = 0.04) were independent adverse predictors of survival. CONCLUSION: AVR for endocarditis carries a relatively high operative mortality, but can result in a satisfactory freedom from recurrent infection, reoperation and long-term survival. Analysis of our series demonstrates that implantation of a biological valve limits the freedom from reoperation and that infection by Staph. aureus reduces the probability of long-term survival.

Surgery for active culture-positive endocarditis: determinants of early and late outcome.

BACKGROUND: The purpose of this study was to describe a single unit experience in the surgical treatment of active culture-positive endocarditis and identify determinants of early and late outcome. PATIENTS AND METHODS: One hundred eighteen consecutive patients with positive blood culture up to 3 weeks before operation (or positive valve culture) and macroscopic evidence of lesions typical for endocarditis, undergoing operation between January 1973 and December 1996 in Southampton, were evaluated. The aortic valve was infected in 53 (48.9%), the mitral in 46 (39%), both aortic and mitral in 12 (10.1%), the tricuspid in 4 (3.9%), and the pulmonary valve in 3 (2.5%). Native valve endocarditis was present in 83 (70.3%) and prosthetic valve endocarditis in 35 (29.7%). Streptococci and staphylococci were the most common pathogens. Mean follow-up was 5.6 years (range, 0 to 25 years). RESULTS: Operative mortality was 7.6% (9 patients). Endocarditis recurred in 8 (6.7%). A reoperation was required in 12 (10.2%). There was 24 late deaths, 17 of them cardiac. Actuarial freedom from recurrent endocarditis, reoperation, late cardiac death, and long-term survival at 10 years were 85.9%, 87.2%, 85.2%, and 73.1%, respectively. On multiple regression analysis the following were independent adverse predictors: pulmonary edema (p = 0.007) and impaired left ventricular function (p = 0.02) for operative mortality; prosthetic valve endocarditis (p = 0.01) for recurrent infection; myocardial invasion by the infection (p = 0.01) and reoperation (p = 0.04) for late cardiac death; and coagulase-negative staphylococcus (p = 0.02), annular abscess (p = 0.02), and longer intensive care unit stay (p = 0.02) for long-term survival. CONCLUSIONS: Operation for active culture-positive endocarditis carries an acceptable mortality. Freedom from recurrent infection, reoperation, and long-term survival are satisfactory. In our data, patients' hemodynamic status at operation was the major determinant of operative mortality. Prosthetic valve endocarditis, coagulase-negative staphylococcus, and annular or myocardial infectious invasion were the critical adverse determinants of late outcome.

Surgical treatment of infective mitral valve endocarditis: predictors of early and late outcome.

BACKGROUND AND AIMS OF THE STUDY: The study aim was to review our experience in surgical treatment of infective mitral valve endocarditis, and to identify predictors of early and late outcome. METHODS: Ninety-one consecutive patients (52 males, 39 females, mean age 55.6 years) underwent surgery between 1973 and 1997 for endocarditis of isolated mitral (n = 65, 71%), mitral and aortic (n = 25, 28%) and mitral, aortic and tricuspid valves (n = 1, 1%). Native valve endocarditis (NVE) was present in 60 patients (66%) and prosthetic valve endocarditis (PVE) in 31 (34%). The main indications for surgery were heart failure in 32 patients, valve dysfunction in 23, vegetations in 21, and persistent sepsis in 11. Eighty-six patients (95%) were in NYHA classes III-IV, and 58 (64%) had active culture-positive endocarditis at surgery. Mechanical valves were implanted in 73 patients and bioprosthetic valves in 13; valves were repaired in five patients. The impact of 46 parameters on early and late outcome was defined by means of univariate and multivariate statistical analysis. Follow up was complete (mean 5.5 years; range: 0-23.1 years; total 507.3 patient-years). RESULTS: Operative mortality rate was 11% (n = 10). Recurrent infection was recorded in five patients (6%), and reoperation was required in eight (9%). Freedom from recurrent infection and reoperation at 10 years was 89.1% and 87.8% respectively. There were 22 late deaths, 15 from cardiac causes. Actuarial survival rates for all patients at 5, 10 and 15 years were 73.0%, 62.7% and 58.7% (for hospital survivors, the corresponding rates were 81.9%, 69.7% and 66.0%). On multiple logistic regression and Cox proportional hazards models, the following were independent predictors: preoperative pulmonary edema (p = 0.01) for operative mortality; PVE (p = 0.02) for recurrence; younger age (p = 0.02) and PVE (p = 0.02) for reoperation; male gender (p = 0.004) and longer ITU stay for survival (if all patients were included); male gender (p = 0.01) and myocardial invasion by infection (p = 0.02) for survival (if only the hospital survivors were analyzed). CONCLUSION: Surgery for infective mitral valve endocarditis carries a relatively high, though acceptable, risk but provides satisfactory freedom from recurrent infection, reoperation and improved long-term survival. Analysis of these data demonstrated that the preoperative hemodynamic status was the major predictor of in-hospital outcome, PVE increased the risk for recurrent infection and reoperation, whereas male gender and myocardial invasion by the infective process critically reduced the probability of long-term survival. The type of offending pathogen, the activity of infection and the involvement of more than one valve did not appear to influence early and/or late outcome.

Prevalence of autoantibodies to ribosomal P proteins in juvenile-onset systemic lupus erythematosus compared with the adult disease.

OBJECTIVE: To determine the prevalence of anti-ribosomal P (anti-P) proteins in several groups of patients with juvenile-onset systemic lupus erythematosus (SLE) in comparison with the prevalence in adult SLE. METHODS: Serum samples were pooled together from 3 cohorts of patients with juvenile-onset SLE in 3 different medical centers and from a miscellaneous group of juvenile-onset SLE patients whose samples were sent by regional physicians. Sera were studied for the presence of anti-P using 2 assays: Western blot with ribosomes as antigen, and an enzyme-linked immunosorbent assay with the COOH-terminal 22 amino acids of the ribosomal P protein in a multiantigenic peptide format as antigen. Sera found positive by both tests were considered positive for anti-P antibodies. Findings from similar studies involving a large cohort of patients with adult-onset SLE from Oklahoma City were used for comparison. RESULTS: The prevalence of anti-P antibodies in the pooled sample of juvenile-onset SLE sera was 45 of 108, or 42%, while in the adult cohort from Oklahoma City, 20 of 260, or 7.7%, were positive for anti-P (odds ratio [OR] 9.6, P < 10(-8) by Fisher's exact test). In addition, it was shown that 12 of 13 patients with both anti-P and anti-double-stranded DNA (anti-dsDNA) in the juvenile SLE cohort had nephritis, while only 8 of 22 patients without both antibodies were nephritic (OR 21.0, P < 10(-8)). It was also shown that in 9 illustrative cases, the levels of anti-P and anti-dsDNA antibodies usually varied together and in concordance with the clinical activity as measured by the SLE Disease Activity Index (SLEDAI). Finally, anti-P-positive and anti-P-negative patients had a similar prevalence of anti-dsDNA, anti-Ro/SSA, and anti-La/SSB antibodies, but patients with anti-P had a higher prevalence of anti-U1 RNP and anti-Sm (P = 0.041 and P = 0.0385, respectively, by Fisher's exact test). CONCLUSION: Antibodies to ribosomal P protein are more prevalent in juvenile-onset SLE than in adult-onset SLE. Levels of antibodies to ribosomal P protein vary with the clinical disease activity as measured by the SLEDAI, often in concordance with the levels of anti-dsDNA. The presence of both anti-P and anti-dsDNA antibodies was powerfully associated with nephritis in the cohort of patients for whom comprehensive clinical and serologic data were available.

Public reaction to Victoria's "2 Fruit 'n' 5 Veg Every Day" campaign and reported consumption of fruit and vegetables.

BACKGROUND: The Victorian "2 Fruit 'n' 5 Veg Every Day" campaign was aimed at increasing awareness of the need to eat more fruit and vegetables and encouraging increased consumption of these foods in the Australian state of Victoria. The demand-side component of the campaign, which had television advertising as a centerpiece, ran from 1992 to 1995. METHODS: Annual postcampaign telephone surveys of approximately 500 Victorians ages 20 and over were conducted with the aim of examining public awareness of the campaign, beliefs about desirable eating habits for fruit and vegetables, and reported consumption of these foods. RESULTS: Over the years, patterns in the level of public awareness, reported consumption, and beliefs about appropriate levels of consumption have tended to parallel changes in the level of mass media investment. During the campaign's most intense period of promotional activity, significant increases in all of these variables occurred. CONCLUSIONS: The results suggest that significant achievements can be made with relatively small-budget mass media promotion of dietary recommendations, especially when part of a more comprehensive program. However, campaigns may need to be adequately resourced for several years if sustained change is to be achieved.

Anti-idiotypic antibodies prevent the serologic detection of antiribosomal P autoantibodies in healthy adults.

A subset of SLE patients has serologically detectable autoantibodies to the ribosomal P proteins (anti-P). We reported the discovery of covert anti-P antibodies and their masking IgG-inhibitory antibodies in the sera of healthy adults. The aim of this study was to determine if these IgG-inhibitory antibodies are anti-idiotypic antibodies (anti-Ids). IgG and IgG-depleted fractions of plasma from two healthy adults were assayed for inhibition of anti-P F(ab')2 binding to the ribosomal P proteins in immunoblot. Anti-P antibody activity was completely inhibited by plasma IgG, whereas there was no inhibition by IgG-depleted plasma. IgG-inhibitory antibodies recognized a cross-reactive epitope among anti-P from different SLE patients. Plasma IgG from one healthy adult was depleted of pepsin agglutinators and generic anti-F(ab')2 antibodies by adsorption with an affinity column prepared with normal IgG F(ab')2. Unretained IgG bound exclusively to anti-P F(ab')2 in ELISA. Using four affinity columns, we isolated IgG anti-Ids to anti-P antibodies from four healthy adults. These purified anti-Ids bound to anti-P F(ab')2 from a healthy adult and SLE patients. They did not bind to F(ab')2 fragments prepared from normal IgG or anti-dsDNA. Ribosomal antigens blocked this anti-Id-Id interaction. Purified anti-Ids inhibited the binding of anti-P F(ab')2 from patients to ribosomal P proteins. SLE patients without overt anti-P antibodies also possessed IgG anti-Ids to anti-P antibodies. We conclude that IgG-inhibitory antibodies are anti-Ids to anti-P antibodies, and are directed to public idiotopes on anti-P antibodies. These anti-Ids may be part of an Id network that regulates anti-P antibody expression, and perhaps pathogenicity.

Development of the anti-ribosomal P autoantibody response.

OBJECTIVE: Autoantibodies to the ribosomal P phosphoproteins (anti-P) appear almost exclusively in SLE. The mechanism by which these autoantibodies appear is unknown. The aim of this study was to determine if IgM and IgG isotype switching occurs during the development of the anti-P autoantibody response. METHODS: Patients who acquired IgG anti-P during clinical observation had their serial serum samples from this period screened for IgM and IgG anti-P by immunoblots and ELISAs. Rabbits were immunized with the immunodominant peptide of the ribosomal P proteins ("P-peptide") cross-linked to bovine serum albumin. Their serial samples were also analyzed for IgM and IgG anti-P before and after repeated immunizations. RESULTS: Two patients were identified who developed transient low titer IgM anti-P that preceded the development of their IgG anti-P. Both patients were clinically well during their peak of IgM anti-P, and developed flares of disease coincident with their high titered IgG anti-P response. The "P-peptide"-immunized rabbits developed an IgM anti-P response that preceded IgG anti-P, and that decreased over time. CONCLUSION: We conclude that IgM to IgG isotype switching occurs in the anti-P autoimmune response. These findings suggest that the autoimmune response to the ribosomal P proteins may be antigen-driven.

The presence of masked antiribosomal P autoantibodies in healthy children.

OBJECTIVE: To determine if antiribosomal P (anti-P) autoantibodies are present in healthy children. METHODS: Sera from healthy children were screened for anti-P by conventional enzyme-linked immunosorbent assay and immunoblot techniques. Sera were also treated with immobilized ribosomal P antigens on nitrocellulose strips; affinity-purified fractions were tested for anti-P by high-sensitivity immunoblot. The relative binding affinities were compared for affinity-purified anti-P antibodies from healthy children and adults, and patients with systemic lupus erythematosus. IgG fractions of anti-P-depleted sera from healthy children were assessed for inhibition of autologous anti-P activity. RESULTS: Conventional serologic screening showed no IgG nor IgM anti-P in 88 untreated sera. IgG anti-P were unmasked in all 79 sera treated by the membrane batch affinity technique. IgM anti-P were identified in 27 of the treated sera; the percentage of positive sera decreased with increasing age (chi(2) for linear trend P = 0.00081). Affinity-purified anti-P from children had relative binding affinities similar to those of anti-P from other groups. Sera from healthy children contained inhibitory IgG antibodies to anti-P. CONCLUSION: These results show that anti-P autoantibodies are present in all healthy children. The majority of these autoantibodies are masked by IgG antibodies, suggesting concordant development of a regulatory network.

Acute infectious arthritis.

Infectious arthritis is one of the few rheumatologic emergencies. Significant morbidity and mortality can occur if treatment is delayed. A high index of suspicion is required when a patient presents with an acute arthritis of one or several joints. Treatment, which is multifaceted, can prevent permanent joint dysfunction.

Anti-ribosomal and 'P-peptide'-specific autoantibodies bind to T lymphocytes.

Patients with systemic lupus erythematosus (SLE) frequently have anti-lymphocyte autoantibodies, some of which also bind to surfaces of neurons. Since anti-ribosomal P protein autoantibodies (anti-P) from SLE patients also bind to surfaces of neurons, we hypothesized that anti-P are anti-lymphocyte antibodies. A panel of human T lymphocytes was evaluated for anti-P binding by indirect immunofluorescence. Affinity-purified anti-ribosomal antibodies were used as a source of anti-P. These autoantibodies bound to the surfaces of all transformed T cell lines tested. This binding was not mediated by Fc receptors. It was inhibitable by ribosomes. Anti-P bound to circulating T lymphocytes from healthy adults and children. They also bound to thymocytes and cord blood T cells from normal neonates. Circulating T cells from SLE patients with anti-P bound less anti-P than cells from healthy controls. Two patients were studied on multiple occasions. The capacity of their T cells to bind anti-P correlated inversely with titres of anti-ribosomal antibodies. Anti-ribosomal antibodies, other than anti-P, also appear to bind to T cells. The surface of T cells contains a protein with the size and antigenicity of the ribosomal P protein, P0. We conclude that anti-ribosomal antibodies are a subset of anti-lymphocyte autoantibodies. Their possible role in the pathogenesis of lymphopenia or lymphocyte dysfunction in SLE has to be defined in further studies.

Development of the anti-Ro autoantibody response in a patient with systemic lupus erythematosus.

OBJECTIVE: To characterize the initial events in anti-Ro production by a patient with systemic lupus erythematosus, in whom this autoantibody is developing. METHODS: The immune response to the Ro ribonucleoprotein and other autoantigens were studied by enzyme-linked immunosorbent assay for IgG and IgM, by isoelectric focusing, and by inhibition studies to determine apparent avidity. RESULTS: The patient's sera showed an oligoclonal response to Ro that increased in complexity and affinity with time. IgM anti-Ro appeared shortly before IgG anti-Ro, and disappeared as IgG anti-Ro increased in titer and affinity. IgG antiribosomal P autoantibodies also appeared during the patient's course, but in contrast to anti-Ro, were not preceded by IgM antiribosomal P. CONCLUSION: These data are consistent with the Ro autoantigen being presented and processed in a manner similar to heterologous antigen, and with differences in the mechanisms that lead to the production of IgG anti-Ro autoantibodies as opposed to antiribosomal P autoantibodies.

Recombinant ribosomal P2 protein can unmask anti-ribosomal P autoantibodies from healthy adults.

Autoantibodies to ribosomal P proteins (anti-P) are detected almost exclusively in the serum samples from patients with systemic lupus erythematosus when conventional enzyme-linked immunosorbent assay and immunoblotting techniques are used. Anti-P are not detected in serum samples from healthy adults by these techniques. By treating serum from healthy adults with ribosome-coated beads, we unexpectedly unmasked anti-P in virtually all individuals. This unmasking of anti-P occurs by the displacement of an antibody inhibitor from anti-P. We wanted to determine whether anti-P from healthy adults could also be unmasked by treatment of their serum or plasma with isolated ribosomal P proteins. Recombinant human ribosomal P2 protein was produced in bacteria as a TrpE fusion protein, resolved by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, blotted onto nitrocellulose membranes, and isolated as strips of membranes corresponding to the size of the P2 fusion protein. Serum or plasma from six healthy adults and three patients with systemic lupus erythematosus were incubated with these strips overnight rather than for 2 hours, as is done in conventional immunoblots. Acid eluates were obtained from the strips and analyzed for antibody activity by immunoblot. Eluates from healthy adults and patients contained antibodies reactive with recombinant ribosomal P2 protein. They also reacted with the three ribosomal P proteins in purified rabbit ribosomes. Their anti-P activity was completely inhibited by a peptide corresponding to the immunodominant linear epitope of the ribosomal P proteins. The antibodies in the eluate were immunoglobulin G. We conclude that anti-P autoantibodies from healthy adults can be unmasked by affinity purification on denatured, recombinant ribosomal P proteins and that antigen excess is sufficient for inhibitor displacement.

Unmasking of anti-ribosomal P autoantibodies in healthy individuals.

Autoantibodies to the ribosomal P proteins (anti-P) are found almost exclusively in a subset of patients with systemic lupus erythematosus. To determine if anti-P autoantibodies are present in healthy individuals but in a masked state, sera from healthy adults were applied to affinity columns coated with ribosomes, and the affinity-purified fractions were analyzed for anti-P Abs by ELISA and immunoblot. Anti-P Abs were detectable in serum only after affinity chromatography, were predominantly of the IgG isotype, stained HEp-2 cells in the characteristic anti-P pattern, and demonstrated specificity for all three ribosomal P phosphoproteins. Using batch affinity chromatography, we identified anti-P autoantibodies in the affinity-purified serum fractions of 54 of 55 healthy individuals. Anti-P Abs from healthy adults generally bound less ribosomal P Ag per mg/ml IgG than anti-P from patients. Inhibition studies revealed that autologous serum contained an IgG inhibitor of anti-P Ab. We conclude that masked anti-P autoantibodies are present in the healthy population and hypothesize that detection of these autoantibodies in systemic lupus erythematosus represents disruption of these regulatory networks.

PIG-A, DAF and proto-oncogene expression in paroxysmal nocturnal haemoglobinuria-associated acute myelogenous leukaemia blasts.

Failed surface expression of the complement decay-accelerating factor (DAF) due to mutation of the PIG-A gene is a hallmark of affected paroxysmal nocturnal haemoglobinuria (PNH) blood elements. Previous findings that acute myelogenous leukaemia (AML) blasts evolving in a PNH patient differed from idiopathic AML blasts in that they exhibited DAF negativity suggested that the leukaemic blasts derived from an affected PNH cell. To investigate whether these cells differ from untransformed PNH cells in PIG-A genetic alterations or in DAF mRNA processing, or are distinguishable from conventional AML blasts in proto-oncogene activation or chromosomal structure, their DNA and RNA were examined using PIG-A, DAF and proto-oncogene probes and their karyotype was analysed. Analyses of the PIG-A genome revealed dual exchanges of A1110-->G and T1130-->A resulting in conversions of T370 to R and I377 to N in the coding region but no deletions or rearrangements. Investigations of DAF mRNA processing showed mRNA species differing in 3' UT regions from those in untransformed cells but similar to those in DAF-positive leukaemia cell lines. Studies of c-myb, c-myc, c-fos and c-fms showed no gross genetic alterations, amplifications or variations in mRNA transcripts deriving from these genes. Karyotypic analysis showed no alterations. The results indicate that in AML blasts evolving in PNH: (1) the PIG-A genome exhibits multiple point mutations but no gross genetic changes; (2) DAF mRNA transcripts exhibit differentiation-dependent variations that do not affect GPI-anchoring; (3) c-myb, c-myc, c-fos and c-fms activation show no differences from idiopathic AML; and (4) no karyotypic abnormalities are associated with AML transformation.