Allergen microarray: comparison of microarray using recombinant allergens with conventional diagnostic methods to detect allergen-specific serum immunoglobulin E

BACKGROUND: The availability of recombinant allergens and recent advances in biochip technology led to the development of a novel test system for the detection of allergen-specific IgE. OBJECTIVE: To test the performance of this allergen microarray in a serological analytical study. METHODS: Standard allergens contained in grass pollen (Phl p 1, Phl p 2, Phl p 5 and Phl p 6) and tree pollen (Bet v 1 and Bet v 2) were used as a model system. The detection of allergen-specific serum IgE using microarrays was compared with standard test systems: CAP/RAST and an in-house ELISA. In order to test the analytical sensitivity of the assays, geometric dilutions of a serum pool containing high levels of pollen-specific IgE from allergic individuals were tested in each system. To assess the analytical specificity, the sera of 51 patients with presumptive allergic symptoms were collected before diagnosis. Thereafter, the results for grass/tree-pollen-specific IgE were compared. RESULTS: The microarray has a good dynamic range similar to the CAP/RAST system. Microarray and ELISA showed comparable analytical sensitivity exceeding the CAP/RAST system. With respect to the analytical specificity, no significant cross-reactivity of the allergens was observed. For two of the allergens tested, weak positive signals were detected in the microarray test system, whereas they were not detectable by CAP/RAST. CONCLUSION: A good correlation of presently used methods to detect serum IgE and the novel microarray test system was observed. As a next step, a careful validation of this method for a multitude of allergens and a thorough clinical evaluation has to be provided. Microarray testing of allergen-specific IgE can be presumed to be the method of choice for a prospective component-resolved diagnosis of Type I allergy, and the basis for the design and monitoring of a patient-tailored specific immunotherapy in the future.     

The presence of messenger RNA for HLA class I in human platelets and its capability for protein biosynthesis

Summary. In order to determine whether platelets contain specific messenger RNA encoding for HLA class I molecules, polymerase chain reaction (PCR) was performed with RNA from different platelet donors. Two amplified 300 bp and 279 bp cDNA fragments were obtained which encompassed sequences from 321 to 620 and from 795 to 1073. The 300 bp fragment encodes exon 2 and exon 3, the 279 bp encodes a portion of exon 4, exon 5, exon 6 and a portion of exon 7. A 300 bp nested PCR product from one donor, that encoded for the highly polymorphic region α2, was cloned and sequenced. The resulting nucleotide sequences fitted to the expected sequence for HLA B*3801 of this donor. Sequence analysis of the 279 bp PCR product demonstrated the presence of exon 5 encoding for the 117 bp transmembrane domain.
In addition, de novo protein biosynthesis was studied by radioimmunoprecipitation of HLA class I molecules from ³⁵S-methionine metabolically labelled platelet lysates with a monoclonal antibody (mab) w6/32 specific for a monomorphic epitope on the heavy chain of HLA class I antigens. Analysis of the immunoprecipitates on SDS polyacrylamide gel electrophoresis showed a specific band with apparent molecular weight (M_r) of 44 kD corresponding to integral membrane HLA protein.
On the basis of these results, we conclude that platelets contain specific messenger RNA encoding for HLA class I molecules and have the capability to synthesize the integral HLA membrane protein.     

Bone-ligament transfer of coracoacromial ligament for acromioclavicular dislocation: A new fixation method used in 6 cases

We treated 6 grade III acromioclavicular injuries with a new fixation method using a bone-ligament transfer of the coracoacromial ligament into a clavicular tunnel. After an average of 16 months, function and cosmesis were excellent in all patients.     

Energy spectra of poly(arylenevinylene)s containing different aromatic units

The energy gaps of poly(p-arylenevinylene)s with phenylene, naphthylene and anthrylene subunits are determined by extrapolating the UV/VIS spectroscopic data of the oligomers. The band structure of the three polymers was theoretically investigated. The topology-, geometry- and correlation-factors were determined upon which the energy spectra of this class of one-dimensional π-electron systems depend.     

Thrombin Inhibition in discordant xenograft rejection

Abstract: Microvascular thrombosis and the associated platelet and endothelial cell activation are prominent observations in xenograft rejection. This pathological picture could be related to the excessive generation of thrombin in the context of either inflammation or putative inter-species molecular incompatibilities between activated coagulation factors and their natural anticoagulants. Relatively selective thrombin Inhibition with the serine protease inhibitor SDZ MTH 958 (MTH-958) are independent of heparinoids and anti-thrombin III. MTH-958 has been shown to significantly prolong porcine cardiac function during perfusion with human blood in an ex vivo model. The aim of this study was to validate the role of thrombin generation in a rodent model of discordant xenograft rejection in vivo. The effect of thrombin inhibition with MTH-958 was tested in both hyperacute rejection (HAR) and delayed xenograft rejection (DXR) after decomplementation with cobra venom factor (CVF) in normal Lewis (Lew) rats and Intrinsic C6 deficiency In PVG (C6-/PVG) recipient rats. Recipient rats received heterotopic guinea pig cardiac xenografts and were treated with titrated doses of MTH-958 until the time of graft rejection. Plasma samples at selected time points were examined to confirm effective thrombin inhibition, and rejected grafts were analyzed by immunohistology. MTH-958 significantly improved graft survival in HAR albeit the extent of prolongation was not marked, but the agent failed to prolong survival In CVF-treated Lew rats. In C6-/PVG rats receiving MTH-958, a significantly reduced graft survival time was observed when compared with C6-/PVG controls. The grafts from MTH-958-treated animals showed dense deposits of C3, IgM, and IgG with fibrin levels similar to controls. The thrombin antagonist tested could prolong xenograft survival during HAR but had no benefit in DXR. The relative non-specificity of the serine protease inhibitor MTH-958 with the potential activation of alternative pathway of complement via the inhibition of factor I could account for the failure to prolong xenograft survival in DXR. The pathogenetic significance of thrombin generation in this situation remains to be determined by the use of more selective and pharmacologically acceptable I anti-thrombin agents.     

Esophagus atresia after cervical spine surgery: case report and literature review

We report on a case of esophageal atresia following cervical spine surgery. A swallowing examination was performed using fibre-optic endoscopy and videofluoroscopy. There was scar tissue fixation of the larynx and esophagus to the cervical spine. Operative mobilization of the larynx and esophagus and formation of a sliding layer using a platysma-fascia flap was done. The PEG and tracheal cannula were removed; oral nutrition was initiated after 3 months. Swallowing disorders following operations on the upper cervical spine should be investigated. Careful preparation that preserves the layers should be carried out. Fixation of tissues as a result of scarring should be treated with a sliding layer.     

Aktuelle und effektive bildgebende Diagnostik von Sprunggelenk und Fuß

Projection radiographic techniques have been used in a standardized manner for decades for the diagnosis of conditions of the foot and ankle; the indications for them and the pattern of findings useful in the initial diagnosis and monitoring of the course are generally known to clinical radiology staff. Computed tomography has been introduced as an extension of the basic procedures performed for diagnosis in the ankle and the hindfoot. In the case of complex fractures, however, specialist experience is essential. CT is an easily accessible investigative procedure and is meanwhile economical and very powerful. Magnetic resonance imaging supplements the range of investigations in special cases when there are particular problems. Microfractures, findings indicative of inflammation and/or dystrophy and, in particular questions concerning the musculoligamentous support system are the situations in which such investigations are needed. Ultrasound has an additive value vis-à-vis magnetic resonance imaging and is an easily accessible and highly effective examination procedure both for the primary diagnosis and for serial monitoring. Financial constraints in the healthcare sector are increasingly limiting purely medical indications. Regardless of this development, the retention of projection radiography, computer tomography, and magnetic resonance imaging must be demanded for the initial diagnosis in the care of foot and ankle injuries. The earliest possible implementation of all these techniques for the initial diagnosis is the first step towards effective treatment. Cafrefully chosen and, if appropriate, complementary strategies are needed for monitoring of the course and for assessment.     

Evaluation of change with time of glomerular morphology in membranoproliferative glomerulonephritis: a serial biopsy study of 33 cases

The evolution of renal glomerular lesions was examined in biopsies taken from 33 patients with membranoproliferative glomerulonephritis (MPGN). 25 patients had a diffuse form of MPGN in the first biopsy (group A). Twenty-four of them still showed diffuse MPGN in subsequent biopsies, but one patient improved clinically and histologically 19 years after the initial biopsy. Out of 6 patients with focal MPGN in the first biopsy (group B), 4 developed diffuse MPGN, one remained with focal MPGN in the repeat biopsy, and another one was found in remission, as determined by both histological and clinical features. Group C represents two patients who had no histological findings of MPGN on initial biopsy but later showed evidence of a diffuse form of MPGN on subsequent biopsies. Thus, the focal form of MPGN may be found either in the development of diffuse MPGN or in its healing stage, and the prognosis will vary accordingly.     

Marked increase of asymmetric dimethylarginine in patients with incipient primary chronic renal disease.

In patients with uremia, increased blood concentrations of the endogenous nitric oxide synthase inhibitor asymmetric dimethylarginine (ADMA) have been linked to the severity of atherosclerosis and to excess cardiovascular mortality. The ADMA levels and several traditional cardiovascular risk factors were assessed in 44 untreated nonsmoking patients with confirmed primary chronic renal disease at different stages of renal disease. True GFR was assessed by means of the inulin-clearance technique. For comparison, nonsmoking subjects matched with respect to age, gender, and body-mass index were examined. Mean plasma ADMA concentration was markedly higher (P < 0.0001) in all patients combined (4.2 +/- 0.9 micromol/L) than in control subjects (n = 16; age 45 +/- 10 yr; serum creatinine 1.0 +/- 0.1 mg/dl; ADMA 1.4 +/- 0.7 micromol/L). However, mean ADMA levels were similar in patients with normal renal function (n = 16; age 41 +/- 9 yr; serum creatinine 1.1 +/- 0.1 mg/dl; GFR 120 +/- 14 ml x min(-1) x 1.73 m2; ADMA 4.0 +/- 0.7 micromol/L), in patients with moderate renal failure (n = 15; 47 +/- 7 yr; 1.8 +/- 0.3 mg/dl; 65 +/- 10 ml x min(-1) x 1.73 m2; 3.8 +/- 0.6 micromol/L) and in patients with advanced renal failure (n = 13; 46 +/- 9 yr; 4.2 +/- 0.9 mg/dl; 25 +/- 4 ml x min(-1) x 1.73 m2; 4.7 +/- 1.2 micromol/L). Furthermore, ADMA levels were increased to the same extent in normotensive (n = 17; 4.0 +/- 0.8 micromol/L) and in hypertensive (n = 27; 4.2 +/- 0.9 micromol/L) patients. In contrast to ADMA, mean total plasma homocysteine concentration were similar in control subjects (10.6 +/- 2.9 micromol/L) and in patients with normal GFR (11.0 +/- 2.9 micromol/L), but were significantly higher in patients with moderate renal failure (17.7 +/- 4.1 micromol/L) and particularly in patients with advanced renal failure (28.2 +/- 10.6 micromol/L). Finally, mean total serum cholesterol concentrations were comparable in the control group and in the three groups of patients with renal disease. In contrast to several traditional cardiovascular risk factors, markedly increased blood concentrations of ADMA, a putative biochemical marker of atherosclerosis, are present even in nonsmoking patients without diabetes with incipient primary renal disease. Thus, the early increase of ADMA levels may be of relevance for the excess cardiovascular morbidity and mortality due to arterio- and atherosclerotic complications in patients with renal disease.