Fieldable genotyping of Bacillus anthracis and Yersinia pestis based on 25-loci Multi Locus VNTR Analysis

<p>Abstract</p> <p>Background</p> <p>Anthrax and plague are diseases caused by <it>Bacillus anthracis </it>and <it>Yersinia pestis </it>respectively. These bacteria are etiological agents for worldwide zoonotic diseases and are considered among the most feared potential bioterror agents. Strain differentiation is difficult for these microorganisms because of their high intraspecies genome homogeneity. Moreover, fast strain identification and comparison with known genotypes may be crucial for naturally occurring outbreaks versus bioterrorist events discrimination.</p> <p>Results</p> <p>Thirty-nine <it>B. anthracis </it>and ten <it>Y. pestis </it>strains, representative of the species genetic diversity, were genotyped by Agilent 2100 Bioanalyzer using previously described Multiple Locus VNTR Analysis assays (MLVA). Results were compared to previous data obtained by standard genotyping system (capillary electrophoresis on automatic sequencer) and, when necessary, direct amplicon sequencing. A reference comparison table containing actual fragment sizes, sequencer sizes and Agilent sizes was produced.</p> <p>Conclusion</p> <p>In this report an automated DNA electrophoresis apparatus which provides a cheaper alternative compared to capillary electrophoresis approaches was applied for genotyping of <it>B. anthracis </it>and <it>Y. pesti</it>s. This equipment, uses pre-cast gels and provides easy transportation, low maintenance and overall general logistic requirements and costs, is easy to set up and provides rapid analysis. This platform is a candidate for on-site MLVA genotyping of biothreat agents as well as other bacterial pathogens. It is an alternative to the more expensive and demanding capillary electrophoresis methods, and to the less expensive but more time-consuming classical gel electrophoresis approach.</p

Passive immunotherapy for N-truncated tau ameliorates the cognitive deficits in two mouse Alzheimer's disease models

Abstract Clinical and neuropathological studies have shown that tau pathology better correlates with the severity of dementia than amyloid plaque burden, making tau an attractive target for the cure of Alzheimer's disease. We have explored whether passive immunization with the 12A12 monoclonal antibody (26–36aa of tau protein) could improve the Alzheimer's disease phenotype of two well-established mouse models, Tg2576 and 3xTg mice. 12A12 is a cleavage-specific monoclonal antibody which selectively binds the pathologically relevant neurotoxic NH226-230 fragment (i.e. NH2htau) of tau protein without cross-reacting with its full-length physiological form(s). We found out that intravenous administration of 12A12 monoclonal antibody into symptomatic (6 months old) animals: (i) reaches the hippocampus in its biologically active (antigen-binding competent) form and successfully neutralizes its target; (ii) reduces both pathological tau and amyloid precursor protein/amyloidβ metabolisms involved in early disease-associated synaptic deterioration; (iii) improves episodic-like type of learning/memory skills in hippocampal-based novel object recognition and object place recognition behavioural tasks; (iv) restores the specific up-regulation of the activity-regulated cytoskeleton-associated protein involved in consolidation of experience-dependent synaptic plasticity; (v) relieves the loss of dendritic spine connectivity in pyramidal hippocampal CA1 neurons; (vi) rescues the Alzheimer's disease-related electrophysiological deficits in hippocampal long-term potentiation at the CA3-CA1 synapses; and (vii) mitigates the neuroinflammatory response (reactive gliosis). These findings indicate that the 20–22 kDa NH2-terminal tau fragment is crucial target for Alzheimer's disease therapy and prospect immunotherapy with 12A12 monoclonal antibody as safe (normal tau-preserving), beneficial approach in contrasting the early Amyloidβ-dependent and independent neuropathological and cognitive alterations in affected subjects

Cyclosporine A in the long-term management of systemic lupus erythematosus

To retrospectively evaluate safety and efficacy of long-term treatment with Cyclosporine A (CSA) in patients with systemic lupus erythematosus (SLE) poorly responsive to treatment with corticosteroids (CCS) and/or conventional disease-modifying anti-rheumatic drugs (DMARDs), SLE patients who had received CSA-based induction and maintenance regimens according to disease activity were recorded. Efficacy was assessed using the SLE Disease Activity Index (SLEDAI) and laboratory analyses. Forty SLE patients [including 18 with lupus nephritis, 11 with neurological involvement and 7 with overlap syndromes (4 Sjögren's syndrome, 2 myasthenia gravis and 1 Behçet's disease)] were recorded. According to baseline SLEDAI, 30 patients had severe and 10 moderate SLE. Mean SLEDAI scores and relevant laboratory values significantly reduced from baseline (22±10 vs 5±6; P < 0.002) during the follow-up period (8±2 years; range 1-15). Twenty-three (57.5%) patients achieved excellent (improvement in the range 70-100%) response to treatment (10 of whom were subsequently maintained on CSA monotherapy), 14 (35%) had good/fair (improvement in the range 25-69%) response and 3 (7.5%) had to interrupt therapy (including CSA) for disease worsening. Mild and transient adverse events occurred in 15 (37%) patients, including hypertrichosis (17.5%), gum hypertrophy (17.5%) hypertension (12.5%), abdominal pain (7.5%), and dyslipidemia (5%), but treatment interruption was not required. Low-dose CSA together with other drugs is effective to induce, or as monotherapy to maintain, long-term (at least 2 years) remission, and is generally well tolerated in patients with moderate or severe SLE poorly responsive to CCS and/or conventional DMARDs. Furthermore, the favourable effect of CSA treatment may allow to spare more cytotoxic drugs. Copyright © by BIOLIFE, s.a.s

Role of antibody-secreting cells as early biomarkers of immune response to influenza vaccination in rheumatoid arthritis patients treated with anti-TNF agents

Evaluation of: Kobie JJ, Zheng B, Bryk P et al. Decreased influenza-specific B cell responses in rheumatoid arthritis patients treated with anti-tumor necrosis factor. Arthritis Res. Ther. 13, R209 (2011). Specific anti-influenza antibodies, early (days 5-7) effector and late (1-6 month) memory B cells have been found to be reduced in immunized rheumatoid arthritis (RA) patients undergoing treatment with TNF-α blocking agents compared with nontreated RA patients and healthy controls. On a per-patient basis, a direct correlation has also been demonstrated between the short-term antibody-secreting effector cells and the 1-month memory B cells, and influenza-specific antibody titer. This study has demonstrated for the first time the reduced specific B-cell and antibody response to influenza vaccine antigens in RA patients undergoing treatment with TNF-α-blocking agents and vaccinated against influenza. This methodological approach, which may allow the identification of early biomarkers, should be followed in order to improve knowledge of immune responses in selected categories of immunosuppressed patients and to modulate immune stimulation intensity and conditions to the level of defective immune response. © 2012 Future Medicine Ltd

Imaging progression despite clinical remission in early rheumatoid arthritis patients after etanercept interruption

The aim of this preliminary study is to evaluate clinical and imaging response in twenty patients with early Rheumatoid Arthritits (eRA) treated with Etanercept (Etn) + Methotrexate (Mtx) and to investigate whether clinical and MRI remission may be maintained after biological therapy interruption. Assessment included: radiography, Visser score and anti-CCP antibodies at baseline; disease activity score in 44 joints (DAS44), rheumatoid factor (RF), Magnetic Resonance Imaging (MRI) of hands and wrists at baseline (T0), 12 (T1), and 24 months (T2). MRI was scored for synovitis, bone oedema and erosions (OMERACT study); patients who reached clinical and imaging remission at T1 were considered eligible for interrupting Etn. At T1 8/20 (40%) patients showed a total remission [DAS44 from 5 (T0) to 1.4 (T1); p&lt;0.02], whereas the other 12/20 (60%) showed an improvement, without complete remission [DAS44 from 4.8 (T0) to 2.8 (T1); p&lt;0.05]. Etn was therefore interrupted in the first group of patients (group A), whereas it was continued in the other group (group B). At T2, group A maintained clinical remission and group B showed further not significant DAS44 reduction from T1. At T1, a significant reduction in synovitis, bone oedema and total score (p&lt;0.01) was observed both in group A and in group B. At T2, group A showed an increase in all the MRI scores that was significant for the synovitis and total score, whereas group B exhibited a further not significant reduction. This preliminary study reports an excellent clinical and imaging response in eRA patients treated with Etn with total remission in 40% of them after a 1-year therapy period. However, it indicates that joint damage may progress, despite a sustained clinical remission, after Etn suspension Copyright © by Biolife, s.a.s

Two-year follow-up in an early rheumatoid arthritis patient treated with etanercept: Radiological involvement despite clinical remission

This case report describes an excellent clinical and radiological response in an eRA patient treated with Etanercept; however, it indicates that joint damage may progress, despite a sustained clinical remission, after Etanercept suspension. Copyright © by BIOLIFE, s.a.s

Health-related quality of life and disability in patients with rheumatoid, early rheumatoid and early psoriatic arthritis treated with etanercept

To assess health-related quality of life (HR-QoL) in patients with Rheumatoid arthritis (RA), early RA and early psoriatic arthritis (PsA), and to evaluate the efficacy of etanercept in reducing disability. Twenty healthy volunteers, 40 RA, 20 early RA and 20 early PsA patients were recruited. All patients received etanercept plus methotrexate. Assessments at baseline and after 2 years' therapy included Disease Activity Scores on 44 joints [DAS(44)], Health Assessment Questionnaire (HAQ) scores and Short Form-36 (SF-36) scores. HAQ and SF-36 scores were significantly worse in patients with RA, early RA or early PsA than in healthy volunteers. The HAQ score at baseline was significantly higher in RA patients than in patients with early RA or early PsA, whereas the scores were similar in patients with early RA and early PsA. Patients with early RA had greater impairment than patients with early PsA in several areas of disability. After 2 years' treatment, HAQ scores and SF-36 summary and subscale scores improved significantly in the three patient groups. This study suggests that early PsA is a less disabling disease than RA or early RA. It confirms the efficacy of etanercept in reducing disease severity and improving HR-QoL and suggests that early therapeutic intervention may lead to greater improvement in the mental and emotional components of these diseases

Efficacy of ivig therapy in inflammatory myositis

The aim of this retrospective study is to evaluate the efficacy and safety of intravenous immune globulin (IVIg) as immunomodulatory treatment in patients with inflammatory myopathies. We analyzed six patients, observed in a single-center from 2004 to 2012, affected by polymyositis (PM), dermatomyositis (DM) or inclusion body myositis (IBM) and treated with IVIg. IVIg has been successfully used as rescue therapy in all patients with a periodical dose of 1-2 g/Kg administered in 5 daysAll patients showed an improvement of muscle strength and a significant reduction of muscle enzyme levels (p<0.01 and p<0.05, respectively). Adverse events were mild and tolerable. Although IVIg has been demonstrated to be effective as steroid-sparing agent when other immunosuppressive drugs had failed, in all patients, however, a progressive reduction of clinical response after a variable number of IVIg cycles was observed. IVIg may safely be used and represents an effective rescue therapy in autoimmune myositis. Larger controlled studies are however needed to establish the IVIg actual efficacy in inflammatory myopathies, define standard therapeutic guidelines and detect predictive factors of respons