Soluble L-selectin increases in the cerebrospinal fluid prior to meningeal involvement in children with acute lymphoblastic leukemia

We have treated 19 B-chronic lymphocytic leukaemia (B-CLL) patients with CDA (Leustat, Janssen-Cilag). Four patients developed severe autoimmune haemolytic anaemia, and 2 of these had severe reticulocytopenia due to red cell aplasia/hypoplasia. Two patients died as a complication of the haemolysis one during the primary episode, with a clinical course suggestive of transfusion associated graft-versus-host disease (taGVHD), and one following a relapse of haemolysis. The onset of haemolysis occurs within 4 cycles of CDA therapy and is temporally related to the T-lymphocyte nadir induced by CDA. The presence of a positive DAT prior to therapy in 3 of 4 patients developing haemolysis suggests that the CDA induced T-lymphocytopenia may exacerbate the tendency of certain CLL patients to autoimmune haemolysis.     

Follow-up study on 24 patients with nomifensine-induced immune haemolytic anaemia

There is as yet only scarce information regarding the natural history and long-term clinical sequelae of patients who survive the acute complications of severe drug-related immune haemolysis, the effect of the mode of drug administration for sensitization and possible alterations of serological characteristics of drug-dependent antibodies during the postsensitization period. We therefore followed 24 patients with nomifensine-induced haemolytic anaemia for up to 6 years after the haemolytic attack. All patients had suffered from a severe haemolytic episode. None of the patients showed any abnormal findings upon reinvestigation (complete history and physical examination; extended biochemical and haematological laboratory status), particularly with respect to renal function. Drug-dependent antibodies remained detectable in 19 of 21 sera, while drug-independent autoantibodies, demonstrable in six patients during the acute phase of haemolysis, could no longer be detected. With regard to the mode of drug administration, the majority of patients (15 out of 24) had developed the haemolysis at the beginning (immediately after a single dose) or during a second course of drug therapy (n = 8 and 7, respectively). The immune response did not appear to be dose- or time-dependent. This study confirms the benign long-term prognosis of patients who survive life-threatening complications in drug-induced immune haemolytic anaemia. In addition, it indicates that drug-dependent antibodies may remain detectable over long periods of time, and that irregular drug administration might be associated with a higher risk of drug sensitization.     

Microangiopathic hemolytic anemia associated with cancers. Symptomatic treatment by plasma exchange

OBJECTIVE: To determine the interest of plasmapheresis in the management of cancer-associated microangiopathic haemolytic anaemia (MHA) not due to cancer treatment. MATERIAL [corrected] AND METHODS: National retrospective study using the file of the French Hemapheresis Society. RESULTS: We isolated 6 patients (5 men and a woman aged 32 to 69-year-old) who had prostatic (4 cases) or breast carcinomas or Hodgkin's disease. Diagnosis of cancer preceeded MHA in 3 patients (from 2 to 4 years). Bone or bone marrow metastases were demonstrated in 5 patients. The clinical presentation included anuria (4 cases), bleeding (4 cases), and laboratory findings were consistent with microangiopathic haemolysis and thrombopenia in all cases and intravascular coagulation (2 cases) and/or renal failure (4 cases). MHA symptomatic treatment included 4 to 10 courses of plasmapheresis, extra-renal epuration (4 patients), anticoagulation (4 patients) and/or antiagregant (3 cases), haemodialysis (4 cases) and vincristine (2 cases). Cancer treatment consisted of antitumoural chemotherapy (2 cases) and/or hormonotherapy (5 cases). In all cases, haemolysis, thrombopenia and intravascular coagulation were controlled within 30 days. MHA treatment was effective alone in 3 patients. No relapse were observed in 3 patients whereas the course of cancer continued in 2 patients. Two patients relapsed and died from MHA after 4 and 36 months. Two patients relapsed and died from MHA within a few months and 1 was lost to follow-up. CONCLUSION: Symptomatic treatment of cancer-associated MHA including plasmapheresis may be useful while waiting for an aetiologic management of the tumour.     

The prevalence of chronic Chlamydia pneumoniae infection as detected by polymerase chain reaction in pharyngeal samples from patients with ischaemic heart disease

AIMS: Cross-sectional serological studies have suggested an association between ischaemic heart disease and infections from Chlamydia pneumoniae and Helicobacter pylori. We therefore sought to find out if patients with ischaemic heart disease had an increased prevalence of C. pneumoniae in the pharynx. As the course of the C. pneumoniae infection remains unclear, both acute and follow-up samples were taken and compared with antibody levels. METHODS AND RESULTS: We studied 282 patients with ischaemic heart disease. One hundred and two subjects without history or symptoms of ischaemic heart disease served as controls. Pharyngeal specimens for polymerase chain reaction detection of C. pneumoniae, and blood samples for C. pneumoniae and H. pylori antibody detection, were collected. In patients with positive polymerase chain reaction or C. pneumoniae IgA titres > or = 32, indicating current infection, convalescent samples were taken at least 6 weeks later. An immunofluorescent antigen detection test was used to confirm the presence of C. pneumoniae elementary bodies in specimens found to be polymerase chain reaction positive. The prevalence of positive polymerase chain reaction tests was 36% among patients and 22% among controls (P<0.05). Forty-seven percent of patients with positive polymerase chain reaction remained positive in the convalescent test. Elevated C. pneumoniae IgG titres > or = 512 were found in 39% of patients and 26% of the controls (P<0.05). IgA titres > or = 32 were found in 46% of the patients and 44% of the controls (ns). Antibody titres remained largely unchanged at convalescent testing. Two patients and none of the controls had IgM titres > 16. There was no link between positive H. pylori serology and positive C. pneumoniae polymerase chain reaction tests. CONCLUSIONS: The high prevalence and persistence of positive pharyngeal C. pneumoniae polymerase chain reaction and elevated antibody titres in patients with ischaemic heart disease indicate a chronic infection. The pharyngeal presence of C. pneumoniae might contribute to a low grade inflammatory activation or be a source for further spread of the bacteria to atherosclerotic vessels.     

Neosporosis and bovine abortion in Scotland

Serum samples were collected over a two-year period from aborting cows and their fetuses from throughout Scotland; 465 maternal sera were examined using an immunofluorescent antibody test for IgG antibody to Neospora caninum and 547 fetal sera were examined for IgM and IgG antibody to the parasite; 355 of the maternal sera were paired with their respective fetal samples. The maternal titres ranged from 1/8 to 1/16,384, with 59 per cent (275) having titres of 1/128 or 1/256. Of the 547 fetal sera, 9.9 per cent had IgM titres equal to or greater than 1/64, 11.2 per cent had IgG titres equal to or greater than 1/64 and 15.9 per cent had IgM and/or IgG titres equal to or greater than 1/64. It is concluded that a fetal IgM or IgG titre of 1/64 or more is evidence of fetal exposure to N caninum but that the examination of maternal sera is less reliable on an individual basis, although valuable for indicating the degree of infection in a herd. Although fetal seropositivity does not necessarily provide proof that N caninum was the cause of death in a given case, the observation that 15.9 per cent of aborted fetuses had antibody to the parasite shows that vertical transmission from dam to fetus is relatively common and may be an important cause of fetal loss in cattle in Scotland.     

Combination therapy with interferon alpha and ribavirin for chronic hepatitis C virus infection in thalassaemic patients

Hepatitis C virus (HCV) infection is common in multi-transfused thalassaemic patients, and, in combination with transfusional iron overload, can result in progressive liver disease. Therapy with interferon-alpha causes a sustained loss of HCV in only 15-25% of patients, and there is as yet no established effective therapy for those who fail to respond. We have conducted a pilot study of combination anti-viral therapy for patients who failed to respond, or relapsed after an initial response to single-agent interferon-alpha. Patients were treated for 6 months with interferon-alpha 2b, given subcutaneously three mega units thrice weekly, together with ribavirin, orally 1 g daily. 11 patients were enrolled, their median age was 24.9 years. 8/10 evaluable patients had cirrhosis on biopsy, five were infected with HCV type 1 and all but one had initial HCV RNA titres > 10(6) genomes/ml. Five patients (45.5%) had a sustained virological response with loss of serum HCV RNA for > 6 months after finishing therapy. There was no clear association between response to therapy and age, histology, HCV genotype, or HCV RNA titre. Transfusion requirements were significantly increased during the treatment phase, probably due to ribavirin-induced haemolysis, and this necessitated intensification of iron chelation therapy. Serum ferritin levels decreased significantly in those who responded. These results suggest that combination therapy is potent in clearing HCV infection, and may provide effective second-line therapy for thalassaemic patients who have failed to respond to interferon-alpha monotherapy.     

Red cell metabolic and membrane features in haemolytic anaemia of alcoholic liver disease (Zieve's syndrome)

Zieve's syndrome (ZS), which consists of transient haemolytic anaemia, jaundice, hyperlipoproteinaemia, and alcohol-induced liver disease, was studied in male patients during the acute (n = 20) and the remittent (n = 10) phase. Chronic alcoholics (n = 10) without haemolysis and healthy male persons (n = 10) served as controls. Erythrocytes were separated into old and young cells by means of density-layer centrifugation. Those fractions which contained older red cells disclosed a pyruvate-kinase instability which resulted in impaired metabolism. Changes in membrane lipid composition as indicated by increased cholesterol and polyunsaturated fatty acids (PUFA) were also detected in patients during the acute phase of ZS. Alcohol-induced red-cell vitamin-E deficiency with a decrease in PUFA levels may provoke an oxidation of reduced red-cell glutathione which in turn results in the enzyme instability. This study lends further support to the hypothesis that the putative role of the red-cell metabolic injury in the origin of haemolysis in ZS cannot be envisaged without introducing membrane-linked and extracellular cofactors.     

Association of Chlamydia pneumoniae IgA antibodies with recently symptomatic asthma

To determine whether recently diagnosed adult-onset asthma is associated with serologic evidence of chronic Chlamydia pneumoniae infection, we performed a case-control study in a primary care clinic of cases with asthma (25 adults reporting first symptoms of asthma within 2 years of enrollment) and 45 concurrently enrolled sex and age (+/- 10 years) matched non-asthmatic controls with normal pulmonary function. C. pneumoniae-specific IgA, IgG and IgG4 antibodies, and circulating immune complexes (CIC) were measured by microimmunofluorescence testing. Results showed that frequencies of IgG titres > or = 16 (92%), IgG4 titres > or = 16 (20%) and CIC > or = 4 (60%) in asthma patients were not significantly different from those of controls. However, asthmatics had a significantly higher prevalence of C. pneumoniae-specific IgA titres > or = 10 (72% of cases vs 44% of controls, P < 0.05). After adjustment for the effects of age, sex and smoking, the odds ratio for an association of IgA and asthma was 3.7 (95% confidence interval 1.2-11.5). We conclude that recently symptomatic reversible airway obstruction in adults is associated with the presence of C. pneumoniae-specific IgA antibodies, a proposed indicator of chronic respiratory C. pneumoniae infection.     

Interaction of per 3,6-anhydro-alpha cyclodextrins (alpha 36CD) and lead-alpha 36CD complex with biological systems

The interactions of per (3,6 anhydro) alpha cyclodextrin (alpha 36CD) and of lead-alpha 36CD complex with biological systems were tested by NMR, ESR and electronic microscopy using erythrocytes and model membranes. It was found that the haemolytic activity of alpha 36CD alone was seven fold lower than that of natural alpha cyclodextrin (evaluated by the concentration inducing 50% haemolysis, DH50 = 35 mM). Conversely, the formation of the complex resulted in an increase of haemolytic properties, with DH50 of 1 mM. The mechanism proposed was an increased membrane diffusion by endocytosis of the complex, leading to higher amounts of intracellular lead.     

Induction of lupus-associated autoantibodies in BALB/c mice by intraperitoneal injection of pristane

Intraperitoneal injection of pristane (2,6,10,14 tetramethylpentadecane) is a standard technique for obtaining monoclonal antibody-enriched ascitic fluid. However, pristane also induces plasmacytomas and an erosive arthritis resembling rheumatoid arthritis in BALB/c mice, probably as a consequence of enhanced interleukin 6 production. We report here that the production of autoantibodies characteristic of systemic lupus erythematosus (SLE) is a further consequence of injecting pristane in BALB/c mice. Anti-Su antibodies appeared as early as 1-2 mo after a single injection of 0.5 ml pristane, followed by anti-U1RNP and anti-Sm antibodies after 2-4 mo. Within 6 mo of pristane injection, 9 of 11 BALB/c mice had developed anti-Su, anti-U1RNP, anti-U2RNP, anti-Sm, and possibly anti-U5RNP antibodies. Autoantibodies were not produced by 20 BALB/c mice of the same age and sex that were not injected with pristane. Thus, autoantibodies characteristic of lupus were induced in mice that are not usually considered to be genetically susceptible to the disease. The induction of autoantibodies associated with SLE by pristane may be relevant to understanding the role of abnormal cytokine production in autoantibody production and the pathogenesis of autoimmune disease. Furthermore, the induction of high titer autoantibodies by pristane dictates caution in the use of ascitic fluid as a source of monoclonal antibodies, since the polyclonal antibodies induced by pristane may copurify with the monoclonal antibody secreted by an injected hybridoma.     

Early effects of ganciclovir therapy on the quantity of cytomegalovirus DNA in leukocytes of immunocompromised patients

The management of patients with autoimmune haemolytic anaemia of warm type (AIHA) is often problematic. Recently, pulsed high-dose dexamethasone (HDD) has been shown to be effective in the treatment of autoimmune thrombocytopenic purpura (AITP). In this study we treated seven patients with AIHA with HDD. The regimen recommended for treatment of refractory AITP (40 mg dexamethasone for 4 d at the beginning of each 28 d cycle) was employed in almost all cases. Prior to dexamethasone administration, haemolysis was decompensated in all seven patients. HDD was well tolerated and led to an improvement of haemolysis in all cases.     

Unclassified haemolytic anaemia with splenomegaly and erythrocyte cation abnormalities--a disease of the spleen?

An unclassified case of haemolytic anaemia with voluminous splenomegaly is reported. This anaemia was normocytic without any specific morphologic aspect of red blood cells (RBC); Coombs test was negative; the osmotic fragility was normal; the increased autohaemolysis was not affected by the presence of glucose; Hb studies were normal; no RBC enzyme deficiency was found; RBC lipids and membrane proteins were normal; there was a marked reduction in RBC survival with exclusive splenic uptake of erythrocytes. Before splenectomy, RBC cations and water content were abnormal: 1) the RBC water was decreased moderately; 2) the RBC sodium was about twice the normal mean with an increased 22Na turn-over; 3) the RBC potassium was markedly reduced and 42K influx was twice the normal mean; 4) the RBC calcium content was increased. Splenectomy was followed by rapid disappearance of haemolysis and RBC water and cation disturbances. Because of this extremely rapid disappearance after splenectomy the authors suggest this case of haemolytic anaemia could be a primary disease of the spleen.     

Chlamydia pneumoniae and acute myocardial infarction in Jerusalem

BACKGROUND: A number of studies in European and North American populations have reported associations between Chlamydia pneumoniae seropositivity and coronary heart disease (CHD). Our objective was to assess the association of IgG and IgA antibodies to C. pneumoniae with acute myocardial infarction (MI) in a population-based case-control study in a Middle Eastern country. METHODS: Eligible cases aged 25-64 with a first acute MI were ascertained through an active surveillance system in Jerusalem hospitals between 1987 and 1989 (85% response). Controls were Jerusalem residents aged 25-64 sampled from the national population registry (83% response). Data on sociodemographic variables, CHD history and risk factors were collected by interview. Chlamydia serology, available for 93% of eligible participants, was performed by microimmunofluorescence on frozen stored samples using the TWAR antigen. Altogether, 251 male and 51 female cases, and 324 male and 162 female controls were analysed. RESULTS: Overall, high IgG titres (> or = 128) were not associated with increased risk of acute MI (Odds ratio [OR] = 0.74 for men [95% confidence interval (CI): 0.47-1.17] and 0.91 for women [95% CI: 0.43-1.94]); neither were high IgA titres (> or = 80) (OR = 1.11 for men [95% CI: 0.71-1.73] and 1.15 for women [95% CI: 0.33-4.0]). At IgG and IgA titres of > 32 and > 20 respectively there was also no relation. CONCLUSIONS: An association of C. pneumoniae seropositivity with acute MI was not confirmed in this population with a very high IgG seropositivity prevalence of 84% in males and 69% in females. However, we cannot exclude the possibility that a postulated recent outbreak obscured an association with chronic C. pneumoniae infection.     

Molecular pathology of ovarian carcinomas

Two patients with lysinuric protein intolerance (LPI) had near-fatal generalized varicella infection with severe interstitial pneumonitis, hepatitis, decreased platelet count, bleeding and hypoalbuminaemia. Active haemolysis resulted in anaemia and massive haemoglobinuria. Serum lactate dehydrogenase activity and ferritin concentration, which in patients with LPI in normal circumstances exceed the upper reference values 3-folds to 10-fold, increased to > 10,000 U/L and > 10,000 micrograms/L, respectively. The patients were treated with fresh frozen plasma, red-cell transfusions and intravenous acyclovir for 14 days, and recovered clinically in a month. Retrospectively, 3 of the 32 other known Finnish patients with LPI had had varicella infection that had been more severe than that in the other children in the family or in subjects in the neighbourhood and had led to hospital admission. Varicella antibodies were measured in 24 patients; 5 had no antibodies and 5 had very low antibody titres. Primary vaccination of three patients with living varicella vaccine increased antibody titres measurably in one patient. We suggest that patients with LPI who have no varicella zoster antibodies should be treated with acyclovir if exposed to varicella and should be (re)vaccinated against chickenpox.     

Serum thyroid peroxidase autoantibodies, thyroid volume, and outcome in breast carcinoma

The prevalence of thyroid peroxidase autoantibodies (TPO.Ab) was assessed in patients with either breast carcinoma or benign breast disease, and its association with disease outcome in breast carcinoma was studied. TPO.Ab were detected by direct RIA in serum from 121/356 (34.0%) of patients with breast carcinoma, compared with 36/194 (18.5%) of controls (P < 0.001); and in 31/108 (28.7%) with benign breast disease, compared with 12/88 (13.6%) of controls (P < 0.05). Survival analysis in a group of 142 women with breast carcinoma demonstrated that TPO.Ab titres > or = 0.3 U/mL were associated with a significantly better disease-free [relative risk (RR) = 1.84, P < 0.05] and overall survival (RR = 3.46, P < 0.02), compared with those who were TPO.Ab-negative. Better outcome associated with higher TPO.Ab titres was confined to those who had thyroid volumes within the intermediate range (10.1-18.8 mL) and did not further enhance the good outcome recorded when volumes were < or = 10.0 mL or > 18.8 mL. Multivariate survival analysis showed that both TPO.Ab and thyroid volume were independently associated with prognosis in breast carcinoma and that RRs for disease-free survival were of a similar order of magnitude to well-established prognostic indices such as axillary nodal status or tumor size. These findings supply evidence that manifestations of thyroid autoimmunity are associated with a beneficial effect on disease outcome in breast carcinoma and provide the strongest evidence to date of a biological link between breast carcinoma and thyroid disease.     

Primary alcohols and phosphatidylcholine metabolism in rat brain synaptosomal membranes via phospholipase D

The prognostic value of maternal serum triple analyte screening with AFP, hCG and uE3 (unconjugated estriol) was studied early in the second trimester of pregnancy. In this case-control study of 38 women and 76 matched controls derived from a consecutive screened population of 28,897, case selection was based upon elevated MSAFP and MShCG (> or = 2 MOM) and low MSuE3 (< or = 0.6 MOM). Adverse pregnancy outcome was found in 65.8% of cases and 2.6% of controls (RR 25, 95% CI 6.3-100.0). When increased odds (> or = 1 in 270) for Down's syndrome were considered with the abnormal analyte screen, fetal/congenital defects, fetal neonatal loss or low birth weight were noted in 17/26 cases (65.4%). Elevated MSAFP and MShCG with low values for estriol, with or without increased odds for Down's syndrome, imply an unfavorable prognosis for both the fetus and the child.     

Differences in haemolytic action of Hg2+ in relation to its concentration and ionic strength of incubating solutions

The haemolytic action of different concentrations of HgCl2 on rat red blood cells (RBC) was studied in vitro. The concentrations of HgCl2 in incubating media were 0.15, 0.25 and 0.50 mmol.l-1. The ionic strength of the media varied from 0 to 154 mmol.l-1 NaCl. Isotonicity of solutions was also compensated using isotonic glucose in different concentrations (287-0 mmol.l-1). Osmolarity of solutions varied from 287 to 308 mOsm. Besides these solutions the haemolysis in Krebs-Ringer solution was also studied. Haemolysis was characterized with two maxima in all concentrations of Hg2+. The first maximum was observed at low ionic strength and the second one at high ionic strength. In relation to the increased concentrations of Hg2+, the first maximum of haemolysis progressively declined towards the higher ionic strength. In the Krebs-Ringer solutions, the increased concentration of Hg2+ was followed by reduced haemolysis. The haemolytic concentration of 0.15 mmol.l-1 was found to be optimal.     

Predictive value of Toxoplasma gondii antibody titres on the occurrence of toxoplasmic encephalitis in HIV-infected patients. ANRS 005/ACTG 154 Trial Group

OBJECTIVE: To study the predictive value of anti-Toxoplasma gondii antibody titres for the occurrence of toxoplasmic encephalitis (TE) in HIV-infected patients. METHODS: Data from the placebo arm of a trial of primary prophylaxis for TE (ANRS 005/ACTG 154) were analysed. Patients included had CD4+ cell counts < 200 x 10(6)/l and a positive Toxoplasma serology. Immunoglobulin (Ig) G and IgM Toxoplasma antibody titres at entry were retrospectively determined by enzyme-linked immunosorbent assay and agglutination on serum samples in a single laboratory. Incidence of TE was estimated by Kaplan-Meier method and a Cox model was used to study the predictive value of antibody titres, adjusted for other covariates. RESULTS: All 164 patients studied were positive for IgG antibodies and one had IgM antibodies. After a mean follow-up of 16 months, 31 cases of TE were documented. One-year incidence of TE was significantly higher in patients with IgG titres > or = 150 IU/ml (23.7%) than in patients with titres < 150 IU/ml (7.7%; relative risk, 3.1; P < 0.003). IgG titres remained significantly associated with the occurrence of TE (relative risk, 3.3; P < 0.005) in the Cox model. Predictive value of IgG titres did not differ according to baseline CD4+ cell counts. CONCLUSIONS: In patients with CD4+ cell counts < 200 x 10(6)/l, IgG anti-Toxoplasma antibody titre is a prognostic factor of occurrence of TE, with a higher risk for titres > or = 150 IU/ml. This finding should reinforce the recommendation of specific prophylaxis in these patients.     

Blood trauma in plastic haemodialysis cannulae

To investigate the haemolysis in haemodialysis cannulae, an in-vitro set up is built, using a unipuncture dialysis system. This system is connected to a bag with fresh calf's blood, by the cannula under test, mounted in a large bloodline (5 mm diameter). The blood characteristics are kept constant by means of a bicarbonate dialysate in the dialyser. During a 6 h period, haematological parameters are regularly samples. Flow through the cannulae is recorded, which is about 500 mL/min. Four different cannulae are tested and compared to the results obtained without any cannula in the circuit. In all cases a linear increase in plasma free haemoglobin levels is found after 6 h. The cannulae can be ranked from 8F catheter over 13G, 14G to 16G cannula, the latter producing the highest degree of haemolysis. When using plastic cannulae at high blood flows, their haemolytic effect may not be neglected.     

Serological titres to Leptospira fainei serovar hurstbridge in human sera in Australia

A set of 723 diagnostic sera from human patients, submitted for the microscopic agglutination test (MAT) for antibodies to a group of 6 leptospiral serovars, was also tested by MAT for antibodies to the recently-discovered Leptospira fainei serovar hurstbridge. MAT titres of > or = 128 to serovar hurstbridge were detected in 13.4% of these sera, and titres of > or = 512 in 7.2%. In contrast, none of 62 sera obtained from a control population of laboratory staff gave titres of > or = 128. The difference between the number of titres of > or = 128 given by the two groups of sera was highly significant (P < 0.01). The titres observed may have been due to cross-reactions with other leptospiral serovars, but this could not be demonstrated. An alternative explanation is that serovar hurstbridge is present in the human population.