A novel assay to measure B cell responses to keyhole limpet haemocyanin vaccination in healthy volunteers and subjects with systemic lupus erythematosus.

The aim of the study was to characterize performance of a complementary set of assays to measure antigen-specific immune responses in subjects immunized with a neoantigen. Healthy volunteers (HV) (n = 8) and patients with systemic lupus erythematosus (SLE) (n = 6) were immunized with keyhole limpet haemocyanin (KLH) on days 1 and 29. Serum antibodies were detected using a flow cytometric bead array (CBA) that multiplexed the KLH response alongside pre-existing anti-tetanus antibodies. Peripheral blood mononuclear cells were studied by B cell ELISPOT. These assays were built upon precedent assay development in cynomolgus monkeys, which pointed towards their utility in humans. Primary anti-KLH IgG responses rose to a mean of 65-93-fold above baseline for HV and SLE patients, respectively, and secondary responses rose to a mean of 260-170-fold above baseline. High levels of anti-tetanus IgG were detected in pre-immunization samples and their levels did not change over the course of study. Anti-KLH IgG1-4 subclasses were characterized by a predominant IgG1 response, with no significant differences in subclass magnitude or distribution between HV and SLE subjects. Anti-KLH IgM levels were detectable, although the overall response was lower. IgM was not detected in two SLE subjects whodid generate an IgG response. All subjects responded to KLH by B cell ELISPOT, with no significant differences observed between HV and SLE subjects. The CBA and B cell ELISPOT assays reliably measured anti-KLH B cell responses, supporting use of this approach and these assays to assess the pharmacodynamic and potential safety impact of marketed/investigational immune-therapeutics.

A retrospective case-control cohort analysis of comorbidity and health expenditure in hospitalized adults diagnosed with obesity utilizing ICD-10 diagnostic coding.

The cost and comorbidity of obesity in hospitalized inpatients, is less known. A retrospective study of patients presenting to a large district hospital in Western Sydney (April 2016-February 2017) using clinical, pathological as well as diagnostic coding data for obesity as per ICD-10. Of 43 212 consecutive hospital presentations, 390 had an obesity-coded diagnosis (Ob, 0.90%), of which 244 were gender and age-matched to a non-obesity coded cohort (NOb). Weight and BMI were higher in the Ob vs NOb group (126 ± 37 vs 82 ± 25 kg; BMI 46 ± 12 vs 29 ± 8 kg/m2 , P < .001) with a medical record documentation rate of 62% for obesity among Ob. The Ob cohort had 2-5× higher rates of cardiopulmonary and metabolic complications (P < .001), greater pharmacologic burden, length of stay (LOS, 225 vs 89 hours, P < .001) and stay in intensive care but no differences in the prevalence of mental disorders. Compared with BMI <35 kg/m2 , inpatients with BMI >35 kg/m2 were 5× more likely to require intensive care (OR 5.08 [1.43-27.3, 95% CI], P = .0047). The initiation of obesity-specific interventions by clinical teams was very low. People with obesity who are admitted to hospital carry significant cost and complications, yet obesity is seldom recognized as a clinical entity or contributor.

Henoch-Schonlein Purpura with thrombocythaemia: an abnormality in Smad4 expression?

We discuss a patient with Henoch-Schonlein Purpura (HSP) and a thrombocythaemia which was diagnosed as a coincidental Essential Thrombocythaemia. We suggest that deficiencies in Smad4 expression may allow for escape thrombocythaemia under the influence of the high levels of TGF-beta found in HSP. With normal Smad4 expression TGF-beta provides inhibition of thrombocyte proliferation. While this needs further elucidation, it could lead to a new approach to classification and management of HSP.

Partial purification of an intercalated disc-containing cardiac plasma membrane fraction.

The sarcolemma of cardiac muscle cells contains a specialised junctional region, the intercalated disc which includes three types of intercellular junction, the macula and fascia adherens and the nexus or gap junction. To facilitate the isolation of these junctions a procedure for the partial purification from mouse hearts of a subcellular fraction containing the intercalated disc region of the sarcolemma was developed. This involved investigating methods of tissue disruption that preserve the integrity of the intercalated disc and minimise myofibrillar entrapment of organelles. Examination of the distribution of marker enzymes showed that relative to the homogenate the intercalated disc fraction prepared by sucrose density centrifugation was only enriched 1.5- to 3-fold in 5'-nucleotidase and (Na+ + K+)-ATPase activities, whereas mitochondrial and sarcoplasmic reticulum marker enzymes were low. The properties of the intercalated disc-containing fraction were compared with the vesicular sarcolemmal fractions devoid of junctional complexes prepared by other methods.

The relationship of anti-DNA antibody idiotypes and anti-cardiolipin antibodies to disease activity in systemic lupus erythematosus.

The detection of anti-cardiolipin antibodies and anti-DNA antibody idiotypes has shown utility in a prospective assessment of 42 lupus patients over a 1-year study period. However, so broad is the range of clinical and serological features included in the diagnostic category of SLE that even a combination of tests will often inadequately reflect disease activity at a given time. For the foreseeable future the value of laboratory investigations will probably lie in supporting clinical judgment of the nature of a patient's illness and the severity of the target organ's dysfunction.

HLA frequency and haplotype analysis in a family study of adult onset rheumatoid arthritis.

Twenty-five families with probands who have rheumatoid arthritis (RA) were studied for clinical evidence of disease and for HLA status. This confirmed an association between RA and DR4 in 19/25 probands (76 per cent, p = 0.008). These 19 probands carried 24 haplotypes which contained DR4. There was no significant increase of DR4 haplotypes bearing B15(Bw62) or B44 when compared with published control haplotype data. The rare complement allele C4 B3 was detected as part of the extended haplotype A2 Cw3 B15(Bw62) DR4 C4 A*3B*3 in three probands with severe RA. Further studies to examine disease severity and autoantibody expression are in progress.

Glycation: a pathological modification in neuropathies?: a hypothesis.

A common feature of a number of neuropathies is the formation of characteristic histopathological lesions of neural amyloid. Although the major components of many of these lesions have been identified, the nature of the modifications of these normal cellular proteins that lead to amyloidogenesis remains elusive. The purpose of this article is to introduce the hypothesis that protein glycation might account for the modifications of normal cellular proteins leading to amyloid formation neuropathogenesis.

Antithrombin III in systemic lupus erythematosus.

Plasma antithrombin III (AT III) was studied in 39 patients with systemic lupus erythematosus (SLE) and in 12 patients with other connective tissue disorders. AT III was measured immunologically by the Mancini method as well as by functional assay using thrombin and the chromogenic substrate, chromozyn TH (Boehringer). Reduced AT III activity was found in 17 patients; 8 had thrombosis. In 6 patients low AT III correlated with disease exacerbations and 2 had systemic vasculitis. No significant correlation could be demonstrated between low AT III levels and thromboembolic disease. A marked variation of functional AT III activity was observed in 30 patients in whom the presence of the lupus anticoagulant was demonstrated. The significance of this association is discussed.

Enhancement of cryoprecipitation from hypotonic serum: a comparative study in normals and in patients with connective tissue diseases.

In a study of 17 normals and 26 patients we have established a normal range for cryoglobulins in a standard and a simple hypotonic system. In a comparison of standard and hypotonic cryoprecipitates in patients with connective tissue diseases we showed a significant increase in protein content (p less than 0.001), IgM (p less than 0.01) and IgM rheumatoid factor (p less than 0.01) in the hypotonic samples. The ratios of IgM RF/IgM in hypotonic cryoglobulins compared to standard cryoglobulins were significantly increased as shown by a chi 2 analysis in both normals (p less than 0.05) and patients (p less than 0.01). Estimation of cryoglobulins in a hypotonic system is a useful simple test which may detect distinct groups of proteins and rheumatoid factors. Precipitates were demonstrated in patients who had previously been considered serologically normal.

Repair of chemical carcinogen-induced damage in DNA of human lymphocytes and lymphoid cell lines--studies of the kinetics of removal of O6-methylguanine and 3-methyladenine.

The removal of O6-methylguanine by human lymphoid cells corresponded, with certain assumptions, to a second-order chemical reaction in any given cell. There was a spectrum of proficiency in this respect for a considerable number of cells originating from different individuals and it was found that patients with diseases associated with autoimmunity tended to fall into the less proficient groups. E-B virus-induced lymphoid cell lines, derived from proficient, but not relatively deficient, peripheral blood lymphocytes, always (in 9/9 cases) reflected the level of proficiency of the donor lymphocytes with respect to removal of O6-methylguanine. Thus while proficient lymphocytes always produced proficient cell lines, deficient lymphocytes, in 3/8 cases, gave rise to more proficient cell lines. No evidence was found that groups of individuals exist who lack ability to remove 3-methyladenine from DNA, either from their blood lymphocytes or derived lymphoid cell lines.

An alternative strategy for therapy of AIDS; a cure?

Present therapeutic strategies in AIDS require continuous treatment. The recognition of the major contribution of an 'autoimmune' component to the pathology of the disease opens up the possibility of ablation of specific T-cell sets as a means of preventing and reversing the disease.

Preliminary study of the relationship between plasma and erythrocyte magnesium variations and some circulating pro-oxidant and antioxidant indices in a standardized physical effort.

The aim of this preliminary study was to determine the relationship between exercise, magnesium (Mg) status, oxidative stress, and antioxidant defence systems. Some corresponding indices have been evaluated: plasma Mg, ascorbate, uric acid, adrenaline, creatine kinase (CK), thiobarbiturate reactive substances, adrenochrome; and in erythrocytes (RBC): reduced and oxidized glutathione (GSH and GSSG) and Mg. Venous blood samples were withdrawn before and 3 min after completion of a 40 min run. Only two significant changes were observed after effort: plasma Mg decreased (P < 0.009) and plasma adrenaline increased (P < 0.005). There was a non-significant tendency for indices of oxidative stress and antioxidant capacity to increase. The significant correlations between plasma Mg and CK (r = -0.88) and between RBC Mg and plasma ascorbate (r = -0.76) disappeared after the effort. Further research is necessary, with a larger number of subjects and variables, to obtain a better understanding of these interactions.

Extraordinary stability of enzymes dried in trehalose: simplified molecular biology.

We show that extremely fragile biomolecules such as DNA restriction and modifying enzymes can be dried in vitro in the presence of trehalose with no loss of activity, even after prolonged storage. A remarkable and unexpected property of the dried enzyme preparations is their ability to withstand prolonged exposure to temperatures as high as +70 degrees C. This stability is unique to trehalose and is not found with other sugars irrespective of their physical or chemical properties. The immediate significance of these observations is the ability to convert enzymes used in molecular biology into stable reagents. The indefinite stability and high temperature tolerance of these dried enzymes should permit the design of convenient formats that may be of particular significance in the automation of genome mapping and sequencing projects. The stabilization of a wide range of biomolecules by trehalose also has practical implications for a number of areas ranging from basic science, through healthcare and agriculture, to bio-electronics.

Inflammatory posterior interosseous nerve palsy in a patient with psoriatic arthropathy.

Psoriasis is a chronic, relapsing, inflammatory skin disorder with a strong genetic basis. Five patterns of psoriatic arthritis have been identified: asymmetrical oligoarticular arthritis, symmetrical polyarthritis, distal interphalangeal arthropathy, arthritis mutilans and spondylitis with or without sacroiliitis. Extra-articular disease is uncommon. We report a rare case of an inflammatory posterior interosseus nerve palsy in a patient with known psoriatic arthropathy, where investigation warranted medical treatment over a surgical approach. The commonest cause of posterior interosseus nerve palsy is entrapment at the proximal forearm. Other possible aetiologies include extension of elbow synovitis as described in rheumatoid arthritis, trauma eg. Monteggia fractures, tumours and iatrogenic injuries. We discuss the diagnostic dilemma and the management issues for upper limb surgeons.