Abbott GmbH Research Articles

Alteration of the Serum N-glycome of Mice Locally Exposed to High Doses of Ionizing Radiation

Exposure of the skin to ionizing radiation leads to characteristic reactions that will often turn into a pathophysiological process called the cutaneous radiation syndrome. The study of this disorder is crucial to finding diagnostic and prognostic bioindicators of local radiation exposure or radiation effects. It is known that irradiation alters the serum proteome content and potentially post-translationally modifies serum proteins. In this study, we investigated whether localized irradiation of the skin alters the serum glycome. Two-dimensional differential in-gel electrophoresis of serum proteins from a man and from mice exposed to ionizing radiation showed that potential post-translational modification changes occurred following irradiation. Using a large-scale quantitative mass-spectrometry-based glycomic approach, we performed a global analysis of glycan structures of serum proteins from non-irradiated and locally irradiated mice exposed to high doses of γ-rays (20, 40, and 80 Gy). Non-supervised descriptive statistical analyses (principal component analysis) using quantitative glycan structure data allowed us to discriminate between uninjured/slightly injured animals and animals that developed severe lesions. Decisional statistics showed that several glycan families were down-regulated whereas others increased, and that particular structures were statistically significantly changed in the serum of locally irradiated mice. The observed increases in multiantennary N-glycans and in outer branch fucosylation and sialylation were associated with the up-regulation of genes involved in glycosylation in the liver, which is the main producer of serum proteins, and with an increase in the key proinflammatory serum cytokines IL-1β, IL-6, and TNFα, which can regulate the expression of glycosylation genes. Our results suggest for the first time a role of serum protein glycosylation in response to irradiation. These protein-associated glycan structure changes might signal radiation exposure or effects.

Immunotherapy in Inflammatory Bowel Diseases

Inflammatory bowel diseases are chronic, relapsing, inflammatory disorders of the intestine. Whereas ulcerative colitis is restricted to the colon, Crohnʼs disease can manifest itself anywhere in the alimentary tract. Ulcerative colitis is characterized by a continuous inflammation of the mucosa and submucosa in the colon. In contrast, Crohnʼs disease frequently affects all layers of the bowel wall and is associated with segmental, discontinuous inflammation. Studies in recent years have highlighted a key role of immunotherapy in inflammatory bowel diseases including treatment with immunosuppressive and biological agents. In particular, treatment with the vav1/Rac1 blocker azathioprine is an established immunosuppressive approach for the treatment of inflammatory bowel disease. This treatment is thought to result in T cell apoptosis with subsequent resolution of gut inflammation in subgroups of patients with inflammatory bowel disease. Furthermore, antibodies against the proinflammatory cytokine TNF have been shown to be effective to induce and maintain remission in patients with these diseases. Adalimumab, infliximab and certolizumab pegol have proven to be particularly effective. Additional findings in recent years have highlighted the role of additional immune mechanisms in the pathogenesis of inflammatory bowel disease. Among these pathways, the IL-6 pathway appears to play a predominant role by controlling epithelial cell proliferation and augmenting T cell resistance against apoptosis. It appears that IL-6 perpetuates chronic intestinal inflammation in patients with inflammatory bowel disease. In addition, IL-6 may boost cancer growth in inflammatory bowel disease, as it directly activates the transcription factor STAT3 in intestinal epithelial cells with subsequent cell proliferation. Several additional immune mechanisms have been identified that appear to contribute to the pathogenesis of inflammatory bowel disease. Among these mechanisms, Th17 cells and their cytokines are potential targets for therapy, although a recent study did not show any benefit of anti-IL-17A therapy in Crohnʼs disease. Furthermore, IL-21 and IL-23 appear to mediate proinflammatory functions, while IL-22 and IL35 seem to suppress chronic intestinal inflammation. Based on the above findings, it is likely that we will see additional new anti-inflammatory and immunosuppressive approaches for the therapy of inflammatory bowel disease in the near future. Conflict of Interest: Lectures and Chairs: Abbott GmbH & Co. KG/Wiesbaden, Falk Foundation/Freiburg, Ferring Arzneimittel/Kiel, Giuliani Pharma SP./Mailand, A. Pentax Europe GmbH/Hamburg Advisor for: Boston Scientific GmbH/Ratingen, Gilead Sciences/Martinsried, GlaxoSmithKline/El Paso, MSD Sharp & Dohme/Haar, Pentax Europe GmbH/Hamburg, ProtAb Ltd./Israel

Abbott GmbH & Co., KG; Abbott Bioresearch Center, Inc.; and Abbott Biotechnology Ltd. v. Centocor Ortho Biotech, Inc., and Centocor Biologics, Inc. [excerpts](United States District Court - District of Massachusetts)Amended Memorandum and Order on Cross-Motions for Summary Judgment

<i>Abbott GmbH &amp; Co., KG; Abbott Bioresearch Center, Inc.; and Abbott Biotechnology Ltd. v. Centocor Ortho Biotech, Inc., and Centocor Biologics, Inc.</i> [excerpts](United States District Court - District of Massachusetts)Amended Memorandum and Order on Cross-Motions for Summary Judgment

A 2-year post-authorization safety study of high-strength pancreatic enzyme replacement therapy (pancreatin 40,000) in cystic fibrosis

Objective: At the request of the Medicines and Healthcare Regulatory Agency and in agreement with the appropriate authorities, an observational, multi-center, non-interventional, post-authorization safety study of high-strength pancreatic enzymes was conducted.Research design and methods: Patients with exocrine pancreatic insufficiency due to cystic fibrosis (CF) who had previously taken high doses of pancreatic enzymes received pancreatin 40,000 capsules (Creon 40,000 Minimicrospheres, Abbott GmbH, Hanover, Germany) as part of their normal treatment for up to 2 years. Initial doses were calculated to match previous established doses in lipase units, with adjustment if required.Main outcome measures: Safety focused on serious suspected adverse drug reactions. Maldigestion symptoms and body weight were also monitored. Patients were managed according to general guidelines common to all major CF units in the UK, although minor variations were expected. The coefficient of fat absorption was not assessed as this was a safety rather than an efficacy study.Results: Sixty-four patients were enrolled at nine UK centers. Two deaths occurred during the study, which were considered unrelated to therapy by investigators. There were no further serious suspected adverse drug reactions related to pancreatin 40,000 and no cases of fibrosing colonopathy. Daily lipase doses were reduced by 11% after switching to pancreatin 40,000. Maldigestion symptoms improved and mean body weight increased from baseline to last observation (mean + 6.1 kg in patients < 18 years old).Conclusions: No safety concerns were identified with pancreatin 40,000 therapy for up to 2 years. Daily lipase doses were not increased when switching to pancreatin 40,000.

An unusual intestinal infection causing intractable diarrhoea of infancy

An eight week-oldmale infant was referred to our hospital with twodayhistory of non-bloodydiarrhoea andaweight lost of 600g ithin two days (more than 10% of body weight). He was the first hild of healthy non-consanguineous parents born spontaneously t termwithaweightof3720ganda lengthof54 cm.Pregnancyand eonatal period were uneventful. Physical examination revealed a atigue infant with a substantial dehydration, reduced skin turgo, o fever, bodyweightof5000g (12thpercentile,Germanreferences rand & Reinken), length of 59.5 cm (82nd percentile) and a head ircumference of 40 cm (57th percentile). Clinical examination neiher revealed organomegaly nor rashes and there was no evidence fneurological disorders onneurological examination.Aparenteral ehydration therapy was started using 0.9% saline solution. The atient developed chronic diarrhoea with 5–14 bowel movements er day. Aiming to restore his homeostasis and replace the masive fluid loss he received between 150 and 390ml/kg/day via aso-gastric tube and parenteral nutrition (PN). Racecadotril 10mg hree times daily (Tiorfan®, Abbott GmbH, Wiesbaden, Germany) nd Lactobacillus GG (Infectodiarrstop® LGG, Infectopharm, Hep-

Rapid remission of psoriasis with reversible leucopenia after two injections with adalimumab

Conflict of interest: none declared. About 20% of patients with psoriasis have severe disease that requires systemic treatment. New forms of treatment such as tumour necrosis factor (TNF)‐α blockers provide a highly targeted approach, thus fewer systemic side‐effects are expected during the course of treatment. However, due to common systemic premedication, laboratory changes may deteriorate during the course of treatment. We report a 34‐year‐old man who developed leucopenia after treatment with adalimumab for severe plaque psoriasis and psoriatic arthritis (Fig. 1a). Leucopenia occurred after two injections of adamimumab 40 mg (Humira®; Abbott Gmbh, Wiesbaden, Germany) given with an interval of 1 week. The patient’s leucocyte level dropped from 3.68 × 109/L to 2.38 × 109/L (normal range 4–11 × 109/L), and neutropenia developed (0.89 × 109/L; normal range 1.5–7 × 109/L). Mild leucopenia (3–4 × 109/L) had been seen previously and was probably related to the long list of previous systemic medications the patient had been taking, which included fumaric acid, leflunomide, methotrexate, ciclosporin and etanercept. Either these treatments had failed to improve his skin or the patient had been unable to tolerate them. The most recent treatment (etanercept) had been discontinued about 9 months before starting adalimumab. Leucocyte levels had been stable for about 12 months at a level of 3–4 × 109/L.

Pitfalls with rapid HIV antibody testing in HIV-infected children in the Western Cape, South Africa

Background Rapid HIV antibody tests are commonly used for HIV diagnosis in the developing world. These tests are generally reported as sensitive, despite paucity of evaluations in paediatric populations. Objectives We tested specimens of paediatric patients, known to be HIV-infected, to detect any false negative tests and determine associations with such an outcome. Study design One hundred and fifty-three specimens, from 109 patients, recorded to be HIV-infected by standard testing, were tested on the Capillus™ HIV-1/HIV-2 test (Trinity Biotech, Ireland); 150 specimens also had sufficient volume to be tested on Abbott Determine™ HIV1/2 assay (Abbott GmbH, Wiesbaden, Germany). Treatment information, CD4 counts and HIV-1 viral load measurements were obtained from patient files and laboratory databases. Results Twenty-one of 153 specimens tested negative on the Capillus (sensitivity 86.3%). False negative results by Capillus were associated with antiretroviral treatment (ART) ( p = 0.0018) and lower HIV-1 viral load ( p = 0.013). Serial dilutions of some of the specimens indicated that both rapid tests, and the Capillus in particular, became negative at lower dilutions than an HIV enzyme immunoassay (EIA). Conclusions The Capillus test had an unexpectedly low sensitivity in a South African population of HIV-infected children that had access to antiretroviral treatment, posing a risk of false negative HIV testing.

Multicenter performance evaluation of a transcription‐mediated amplification assay for screening of human immunodeficiency virus‐1 RNA, hepatitis C virus RNA, and hepatitis B virus DNA in blood donations

The performance of the recently launched Procleix Ultrio (Chiron/Gen-Probe) human immunodeficiency virus-1 (HIV-1), hepatitis C virus (HCV), and hepatitis B virus (HBV) blood screening assay was evaluated in a European multicenter study. Serial dilutions of reference materials were tested to determine the detection limits. Robustness and specificity were assessed by testing alternating high-load HCV RNA-positive and -negative samples, and 2912 test pools of eight donations. The added value of minipool and single-donation HBV nucleic acid testing protocols was compared to the currently used Prism (Abbott GmbH & Co. KG) hepatitis B surface antigen (HBsAg) and Auszyme (Abbott GmbH & Co. KG) dynamic HBsAg tests in 15 HBV seroconversion panels. The 95 percent detection limits (and 95% confidence interval [CI]) on the WHO International Standards was 26 (16-58) IU per mL for HIV-1 RNA, 4.6 (3.7-6.5) IU per mL for HCV RNA, and 11 (7.3-22) IU per mL for HBV DNA. No cross-contamination was observed. Testing 2912 pools of eight donations revealed 16 initial reactive samples; 11 were confirmed. The specificity after initial testing and percentage of invalid results were 99.83 and 0.48 percent, respectively. The HBV window-period (WP) reductions relative to HBsAg seroconversion in Prism and Auszyme dynamic HBsAg were, respectively, 6 days (95% CI, 3-8) and 9 days (95% CI, 7-12) in 1:8 minipool (MP) testing. The performance characteristics of Procleix Ultrio assay and the Procleix HIV-1 and HCV assay are comparable. The sensitivity for HIV-1 and HCV met the directives of the Paul-Ehrlich Institute and the FDA. The assay can reduce the WP for HBV by 6 days to 2 weeks when used in small MP (<1:8) or single-donation screening protocols.

Randomized Clinical Trial of Itopride for the Treatment of Postoperative Ileus after Laparoscopic Cholecystectomy

Background. Postoperative ileus is a distressing and frequent adverse effect after laparoscopic cholecystectomy (LCE). The goal of this prospective study was to evaluate if the new prokinetic drug itopride hydrochloride (Ganaton®, Abbott GmbH Co. KG, Wiesbaden, Germany) may ameliorate the gastric motility in the early post-operative period after LCE. Transcutaneous electrogastrography (EGG), a noninvasive diagnostic method allowing a monitoring of gastric myoelectric activity, was used to evaluate changes of gastric motility. Methods. 50 patients undergoing LCE were observed for 3 days beginning with the day of surgery. The patients were perorally treated with itopride hydrochloride (50 mg per day) or placebo (sacharose) in a randomized double-blind manner. EGG records were done 6, 24, and 48 h after surgery. EGG was performed in a fasting state and after stimulation with a liquid bolus. The EGG data were recorded by a Microdigitrapper device and analyzed using spectral analysis and Fourier transformation. Results. When comparing both groups (itopride and placebo), nausea was found significantly more frequently in the placebo group at the day of surgery as well as on postoperative days 1 and 2. The incidence of vomiting did not differ between both groups. Moreover, there are some differences in the EGG records. On the day of surgery, 40 and 56% of the patients in the placebo and the itopride group showed a physiological EGG curve, respectively. On the 1st postoperative day it was 56 and 68% and on the 2nd postoperative day 80 and 88%. However, these differences failed to reach a significance level of p < 0.05, most likely because of the relative small group sizes. Conclusion. The perioperative use of itopride accelerates the normalization of the EGG curve after LCE. Itopride was well tolerated, and we did not observe serious adverse effects during therapy. Our data suggests that itopride can be a useful accelerator of postoperative ileus restoration after LCE. However, other studies will be necessary to unequivocally prove the beneficial effect of itopride after LCE also in comparison with other commonly used drugs.

Evaluation of newly developed microparticle enzyme immunoassays for the detection of HCV antibodies

The newly developed anti-HCV assays AxSYM HCV version 3.0 and IMx HCV version 3.0 were evaluated with regard to their precision, sensitivity and specificity in comparison to the HCV EIA 3.0 (Abbott GmbH, Wiesbaden, Germany). Precision testing was undertaken using five positive controls with different anti-HCV levels for each assay. Specificity was estimated by testing 4383 blood donor specimens. The supplemental assay Matrix HCV 2.0 (Abbott GmbH, Wiesbaden, Germany) was used to confirm repeatedly reactive results. Samples which had been found to be positive or indeterminate by Matrix HCV 2.0 were tested by qualitative polymerase chain reaction after reverse transcription (RT-PCR, Amplicor™ HCV test, Roche Diagnostic Systems, Basel, Switzerland). To determine sensitivity, 20 commercially available seroconversion panels were tested. Based on supplemental testing, the apparent specificities of AxSYM HCV version 3.0, IMx HCV version 3.0 and HCV EIA 3.0 were estimated to be 99.84, 99.98 and 99.80%, respectively. In seroconversion panel testing, AxSYM and IMx HCV version 3.0 detected seroconversion in up to 12/20 panels earlier and in up to 1/20 cases later than the comparison EIA. The highest sensitivity was shown in AxSYM HCV version 3.0, followed by IMx HCV version 3.0 and HCV EIA 3.0. Based on the improved seroconversion sensitivity and specificity, the AxSYM and IMx HCV version 3.0 assays appear to be suitable for detecting HCV antibodies in blood donor testing and other routine laboratory assessments.

Feasibility and efficacy of routine PCR screening of blood donations for hepatitis C virus, hepatitis B virus, and HIV-1 in a blood-bank setting

Despite sensitive antibody-based blood-donor screening, a residual risk of transfusion-transmitted viral infections exists. Only direct monitoring by sensitive nucleic-acid tests would provide data accurately to measure the risk and to assess risk-reduction procedures. We investigated the feasibility and efficacy of routine screening of donors for hepatitis C virus (HCV), hepatitis B virus (HBV), and HIV-1 by PCR. For PCR testing, individual donor plasma samples were pooled (96x100 microL) overnight by two automatic pipetting machines. Viruses were concentrated by centrifugation and nucleic acids were extracted. HCV PCR was done on the Cobas Amplicor system (Hoffmann-La Roche, Mannheim, Germany). HBV and HIV-1 sequences were amplified by single (non-nested) in-house PCRs and detected by agarose-gel electrophoresis. Detection limits were 1000-5000 genome equivalents/mL in the donor blood. PCR testing was done in parallel to antibody screening with a maximum throughput of 3000 samples in 7-8 h. Positive samples were identified 1-2 days later. 111 of 373,423 donations (107 of 4500 pools) were PCR and antibody/antigen-confirmed positive. We found one HCV PCR-positive antibody-negative donation with normal alanine aminotransferase and one HCV PCR-positive donation with an elevated alanine aminotransferase (100 IU), which was negative in the AxSYM 2.0 and Matrix 1.0, but positive after control in the Abbott Prism test (Abbott GmbH, Wiesbaden, Germany). PCR is a suitable and fast blood-donor screening procedure and contributes to a reduction in viral transmission by transfusion of blood components. In our selected donor population, the yield of detected contaminated donations from donors in the time window in which they are highly infectious but do not have any symptoms or detectable antigen and antibody concentrations (diagnostic window), confirms theoretical estimates.