Recominant Expression and Immunological Characterization of House Dust Mite Allergen Der P 1

The cysteine protease Der p1 from dust mite of the genus Dermatophagoides pteronyssinus is a major type I allergen. About 80% of house dust mite (HDM) allergic individuals are reactive to this protease in standard assays for detection of IgE. A curative treatment for atopic allergy is immunotherapy (IT) with HDM extracts which are complex mixtures occasionally resulting in anaphylactic reactions. Novozymes focuses on developing a recombinant variant of Der p1 which exhibit lowered risk of IgE-mediated allergic reactions, while maintaining its ability to trigger proper Th-cell responses. This may provide a safer alternative for specific IT of HDM allergy. A secreted recombinant form of pro-Der p 1 expressed by Saccharamyces cerevisiae was obtained by fusion of the pro-enzyme to a fungal signal peptide. The N -glycosylation site of Der p1 was mutated resulting in a deglycosylated pro-enzyme with a molecular mass of 35 kDa. Protein purification procedure was developed to obtain nearly pure Der p1 protein followed by determination of concentration by active-site-titration with the cysteine protease inhibitor E64. The deglycosylated recombinant pro-Der p 1 revealed immunologic similarity to the native Der p 1 molecule when compared in basophile histamine release, IgE-binding assays and T-cell proliferation assays. By in silico epitope mapping of a modelled 3-dimensional structure of Der p1, five putative IgG and IgE epitopes were predicted. By protein engineering, the predicted epitopes were removed one by one in Der p1 and screening for hypoallergenic variants was performed.     

Fetal Plasma Concentrations after Intraamniotic Sufentanil in Chronically Instrumented Pregnant Sheep

Background: Rapid progress is being made in fetal surgery. Because the fetus is capable of pain perception after the 26th week of gestation, adequate postoperative fetal pain management is essential. The preferred approach would provide fetal analgesia without affecting the mother. Intraamniotically administered sufentanil may be an interesting option if it achieves therapeutic plasma concentrations (PCs) in the fetus but not the mother.

Methods: After approval of the study, 25 or 50 [mu]g sufentanil was administered intraamniotically in 10 chronically instrumented pregnant ewes. Maternal and fetal vital signs, arterial blood gases, and uterine blood flow were recorded over 120 min. Sufentanil PCs were determined before and 1, 3, 5, 10, 15, 30, 45, 60, 90, and 120 min after injection. Statistical analysis was performed using one- or two-way analysis of variance followed by Dunnett or Tukey test, as appropriate (P < 0.05; data presented as median [95% confidence interval]).

Results: After 25 [mu]g sufentanil, fetal PC stabilized at 134 +/- 89 pg/ml (after 10 min), and maternal PCs stabilized at 44 +/- 11 pg/ml (after 15 min). After 50 [mu]g sufentanil, fetal PCs stabilized at 134 +/- 35 pg/ml (after 15 min), and maternal PCs reached 80 +/- 25 pg/ml (at 30 min). Injection of 25 [mu]g sufentanil intraamniotically did not affect maternal or fetal hemodynamics, uterine blood flow, or arterial blood gases. Fetal heart rate increased after administration of 50 [mu]g sufentanil (maximum change at 10 min: +16 +/- 12%).     

Visual neglect in right-sided apoplectic lesions

Fifty-five patients with right-sided apoplectic brain lesions prospectively and consecutively collected, were analyzed as to the frequency of visual neglect. The syndrome was present in 18 patients, 10 of whom had left hemi- or quadrantanopia. In addition, two patients had anopia with Anton's syndrome.
Only the visual neglect without visual field defects was closely related to denial of hemiparesis.
The two components of the syndrome of visual neglect, defects in registering visual information from the left side and defective appreciation of own condition are described and analyzed.
Psychological theories of the processes behind the syndrome of neglect have difficulties in joining the cardinal features of the syndrome satisfactorily. Theories on the specific role of the right hemisphere in connection with attention/arousal and complex gnostic functions are referred to in the discussion of the syndrome.     

Prevalence of Retinopathy and Proteinuria in Type 1 Diabetics in Iceland

ABSTRACT. The prevalence of retinopathy and proteinuria was assessed in 212 and 230 type 1 diabetics in Iceland respectively. They represent 78% and 84% of all such patients identified in the country. Retinopathy was present in 33.5%, background lesions only in 27.4% and more severe retinopathy in 6.1%. After 5–9 years of diabetes (mean 6.9) the prevalence of retinopathy was 18.8%, including 2.1% proliferative, and rose to 76.7% after 20 years or more of diabetes (mean 26.7), including 16.2% in the proliferative stage. Blindness was found in 2.4% of the patients. Diabetics diagnosed at the age of 0–19 years had a lower prevalence of retinopathy during their first 20 years of diabetes than those diagnosed later in life (p<0.05). Proteinuria was present in 14%, intermittent in 10% and continuous in 4%. After 10 years or more of diabetes (mean 19.3) the prevalence of intermittent and continuous proteinuria was 13.0% and 8.3%, respectively.     

Malignant melanoma in pregnancy. A population-based evaluation

BACKGROUND: For many years, there has been controversy in the medical community regarding the correlation of female hormonal factors with the outcome of women with malignant melanoma. There have been multiple reports that women with high hormone states, such as pregnancy, had thicker tumors and/or a worse prognosis compared with a group of control women. METHODS: The authors used a database that contained maternal and neonatal discharge records from the entire state of California from 1991 to 1999 and linked those records to the California Cancer Registry, which maintains legally mandated records of all cancers reported in California during the same time period. Four hundred twelve women with malignant melanoma diagnosed during or within 1 year after pregnancy were identified (145 antepartum, 4 at delivery, and 263 postpartum) and were compared with a group of age-matched, nonpregnant women with melanoma (controls). The database captured only pregnancies at > or = 20 weeks of gestation. RESULTS: When comparing women who had pregnancy-associated melanoma with the control group, the authors found no difference in the distribution of disease stage (82.0% of pregnant and postpartum women had localized melanoma vs. 81.9% of control women) or the tumor thickness (mean: 0.77 mm for pregnant women, 0.90 mm for postpartum women, and 0.81 mm for the control group). In a multiple regression model that controlled for age, race, stage, and tumor thickness, pregnancy had no impact on survival in women with melanoma. Lymph node assessment and positivity of lymph nodes also were equivalent between the two groups. Maternal and neonatal outcomes did not differ between pregnant women with melanoma and control women who were pregnant and had no history of malignancy. Small numbers of women with advanced melanoma and the inability to capture melanoma that occurred in pregnancies that were lost or were terminated prior to 20 weeks limited the conclusions primarily to women with localized melanoma. CONCLUSIONS: In this large, population-based study of pregnant women in California from 1991 to 1999 with malignant melanoma, there were no data found to support a more advanced stage, thicker tumors, increased metastases to lymph nodes, or a worsened survival. The outcome for women with localized melanoma associated with pregnancy was excellent. Maternal and neonatal outcomes also were equivalent to those of pregnant women without melanoma.     

Reorganization and orientation of regenerating nerve fibres,perineurium, and epineurium in preformed mesothelial tubes – an experimental study on the sciatic nerve of rats

Regeneration of severed peripheral nerves is unfortunately often incomplete, due to loss of nerve fibers and neuroma formation. A new approach is presented with the intention of improving the conditions for nerve repair. In the first of the two stages, a pseudosynovial tube is formed around a silicone rubber rod, surrounded by a stainless steel spiral, which was placed in the backs of rats. This tube, in the second stage, is used as a free “tube graft” to bridge gaps of about 10–12-mm lengths in the severed sciatic nerve. The tube was kept open by the metal spiral. Regenerating nerve fibers with their sprouts grew into the initially open space in the tube. A new nerve trunk was formed, comprised of closely packed myelinated and unmyelinated axons, organized into fascicles. Demonstration by electron microscopy and by EMG recording of reinnervation of foot muscles supported successful long-term results. The fascicles were delimited by perineurial and epineurial sheaths and, furthermore, showed signs of maturation. It was also demonstrated that the nerve-fiber regeneration ceased after a few weeks if there was no distal nerve inserted into the tube. The importance of optimizing the interaction between local factors and regenerating nerve fibers for reestablishment of functionally valuable motor units is discussed.     

Torture sequelae located to the skin

Torture sequelae and associated diagnostic problems within the field of dermatology are reported. Patient 1 claimed to have been subjected to torture in 1973 in South America and reported that he had a thick rubber-like rope tied around each thigh just above the knees. Six years later a linear zone of alopecia was observed extending circularly around each thigh, probably representing cicatricial alopecia. Patient 2 claimed to have been subjected to torture in November 1978 in Iraq and reported that he was burned on several areas of the skin with a metal rod of the size and shape of a cigarette. One year later, several circular or serpiginous scars with a markedly atrophic centre and a narrow hypertrophic, distinctly demarcated peripheral zone were observed on the skin. They probably represent the sequelae to deep third-degree burns produced by an electrically heated, cylindrical metal rod. Patient 3 claimed to have been subjected to torture in July and August 1974 in India. She reported to have been burned on several areas of the skin with a cigar as well as lifted by her hair. A few days after the torture she developed transient features typical of traction alopecia in the scalp, including the appearance of pimples. The examination 6 years later revealed on the extremities two irregularly shaped and indistinctly demarcated scars, one of them slightly atrophic, which might be sequelae to burns produced by an ember from a cigar.