Immunological characterization of Asp f 2, a major allergen from Aspergillus fumigatus associated with allergic bronchopulmonary aspergillosis

The 37-kDa recombinant protein Asp f 2, encoding an allergen of Aspergillus fumigatus, was expressed in a prokaryotic expression system and immunologically evaluated for its functional and structural properties. The open reading frame for a 310-amino-acid-long protein was shown to encode a signal peptide of 31 amino acids. A native 37-kDa culture filtrate protein and a 55-kDa mycelial glycoprotein (gp55) exhibited complete N-terminal sequence homology to Asp f 2. A GenBank search for homologous proteins revealed 60 and 44% sequence homologies to the cytosolic protein ASPND1 from Aspergillus nidulans and fibrinogen binding protein from Candida albicans, respectively. The glycosylation sites and cysteine molecules are conserved in all the three proteins. The extracellular matrix protein laminin showed a dose-dependent interaction with Asp f 2. This protein, expressed as a major cell-associated protein within 24 h of in vitro fungal culture, comprises 20 to 40% of total fungal protein. Furthermore, both native and recombinant Asp f 2 exhibited specific immunoglobulin (IgE) binding with allergic bronchopulmonary aspergillosis (ABPA) and cystic fibrosis-ABPA patients, whereas A. fumigatus-sensitized allergic asthma and normal control subjects failed to show IgE binding with Asp f 2. These results indicate that Asp f 2 is a major allergen of A. fumigatus exhibiting IgE antibody binding with sera from patients with ABPA. The antigen should be explored further for its potential role in the differential diagnosis of A. fumigatus-associated allergic diseases.     

FISH-based analysis of stable translocations in a Techa River population

Bone morphogenetic proteins (BMPs) and their receptors (BMPRs) are thought to play an important role in bone morphogenesis. The purpose of this study was to determine the locations of BMP-2/-4, osteogenic protein-1 (OP-1, also termed BMP-7), and BMP type II receptor (BMPR-II) during rat fracture healing by immunostaining, and thereby elucidate the possible roles of the BMPs and BMPR-II in intramembranous ossification and endochondral ossification. In the early stage of fracture repair, the expression of BMP-2/-4 and OP-1 was strongly induced in the thickened periosteum near the fracture ends, and coincided with an enhanced expression of BMPR-II. On day 7 after fracture, staining for BMP-2/-4 and OP-1 immunostaining was increased in various types of chondrocytes, and was strong in fibroblast-like spindle cells and proliferating chondrocytes in endochondral bone. On day 14 after fracture, staining with OP-1 antibody disappeared in proliferating and mature chondrocytes, while BMP-2/-4 staining continued in various types of chondrocytes until the late stage. In the newly formed trabecular bone, BMP-2/-4 and OP-1 were present at various levels. BMPR-II was actively expressed in both intramembranous ossification and endochondral ossification. Additionally, immunostaining for BMP-2/-4 and OP-1 was observed in multinucleated osteoclast-like cells on the newly formed trabecular bone, along with BMPR-II. In reference to our previous study of BMP type I receptors (BMPR-IA and BMPR-IB), BMPR-II was found to be co-localized with BMPR-IA and BMPR-IB. BMP-2/-4 and OP-1 antibodies exhibited distinct and overlapping immunostaining patterns during fracture repair. OP-1 may act predominantly in the initial phase of endochondral ossification, while BMP-2/-4 acts throughout this process. Thus, these findings suggested that BMPs acting through their BMP receptors may play major roles in modulating the sequential events leading to bone formation.     

Mechanism of ovalbumin proteolysis

Papain, thermitase and pepsin at pH 4.0 hydrolyze ovalbumin to low molecular weight peptides. Simultaneously a stepwise formation of several short fragments resulting in the appearance of a group of high molecular weight fragments and a subsequent splitting of the latter into two fragments of intermediate molecular mass takes place. The fragments are retained in the molecule by noncovalent bonds. The fragments formed under the action of different enzymes are similar in size, which is suggestive of splitting in the same narrow segments of the ovalbumin polypeptide chain. The results obtained indicate that during proteolysis of the native ovalbumin two extreme types of the proteolysis--the one-by-one and zipper types--occur in parallel, while the course of the latter is largely determined by the ovalbumin structure.     

Ketorolac tromethamine in the maintenance of intraoperative mydriasis

BACKGROUND AND OBJECTIVE: Mydriatics and antiprostaglandins are routinely applied preoperatively to facilitate the aspiration of cortical material during extracapsular cataract surgery and to prevent intraoperative miosis. Unfortunately, the ideal antiprostaglandin has yet to be introduced. This study represents the first prospective, case-controlled, randomized, masked, clinical trial comparing the efficacy of oral ketorolac tromethamine (KT) with the efficacy of topical flurbiprofen sodium (FS) and a placebo (normal saline) in maintaining intraoperative mydriasis without adrenaline in the irrigation solution. PATIENTS AND METHODS: Sixty-three patients, undergoing extracapsular cataract extraction with posterior chamber intraocular lens implantation, were randomly divided into three equal groups. Group 1 received 20 mg of oral KT 2 hours prior to surgery, group 2 received 0.03% FS topical solution at the recommended dosage, and group 3 received topical normal saline in a regimen identical to that of group 2. Identical mydriatics were used in both groups. The patients' pupils were measured at five stages of surgery. RESULTS: The mean pupillary diameters at each surgical stage were greater in group 1 than in group 2; however, the difference was not statistically significant. The pupils in group 3 were significantly smaller than those of group 1 or group 2 (P < .05). No systemic toxicity to KT was observed. Moreover, the need for additional postoperative analgesics was significantly greater in groups 2 and 3. CONCLUSIONS: Oral KT is safe, convenient, and as effective as topical FS, and has the added advantage of reducing postoperative pain.     

Myocardial revascularisation in elderly patients with refractory or unstable angina and advanced coronary disease

BACKGROUND: Elderly patients with ischaemic heart disease are often treated more conservatively and for longer than younger patients, but this strategy may result in subsequent invasive intervention of more advanced and higher risk coronary disease. METHODS: We performed a retrospective analysis of 109 patients aged > or = 70 years (mean age 74 years, 66% men), who presented with angina refractory to maximal medical treatment or unstable angina over a 2-year period (1988-1990), to compare the relative risks and benefits of myocardial revascularisation [coronary artery bypass grafting (CABG) and percutaneous transluminal coronary angioplasty (PTCA)] in this higher-risk age group. RESULTS: Sixty patients underwent CABG and 49 patients PTCA. There were eight periprocedural deaths in total (six in the CABG group, and two in the PTCA group, P = 0.29). Six patients in the CABG group suffered a cerebrovascular accident (two fatal). Acute Q-wave myocardial infarction occurred in one patient in the CABG group and in two patients in the PTCA group. The length of hospital stay was longer for the CABG group (CABG group 11.4 +/- 5.4 days, range 7-30 days, PTCA group 7.4 +/- 7.6 days, range 1-39 days, P = 0.01). Outcome was assessed using the major cardiac event rate (MACE; i.e. the rate of death, myocardial infarction, repeat CABG or PTCA). The cumulative event-free survival in the CABG group in 1, 2 and 3 years was 87, 85 and 85%, respectively. In contrast, in the PTCA group it was 55, 48 and 48% (P = 0.0001). Age, sex, number of diseased vessels, degree of revascularisation and left ventricular function were not predictive of the recurrence of angina in both groups. Actuarial survival (total mortality, including perioperative mortality) was lower at 1 year in the CABG group due to the higher perioperative mortality, but similar in both groups after the second year (P = 0.62). CONCLUSIONS: Elderly patients with refractory or unstable angina who are revascularised surgically have a better long-term outcome (less frequent event rate of the composite end-point--myocardial infarction, revascularisation procedures and death) compared with those who are revascularised with PTCA. This benefit is been realised after the second year. Total mortality is similar in both groups after the second year. Therefore elderly patients who are fit for surgery should not be denied the benefits of CABG. PTCA may be regarded as a complementary and satisfactory treatment, especially for those whose life expectancy is limited to less than 2 years. The use of stents may improve outcome in the PTCA group and this needs to be evaluated.     

Central nervous system cryptococcosis: parenchymal calcification and large gelatinous pseudocysts

In an 11-year-old immunocompetent girl with protracted cryptococcal infection of the central nervous system, CT showed multiple areas of parenchymal calcification. MR imaging showed large gelatinous pseudocysts around the brain stem. These imaging features and the child's age are unusual for intracranial cryptococcosis.     

Ceftriaxone in treatment of streptococcal endocarditis

The authors studied efficacy and safety of ceftriaxone given intravenously for 3-4 weeks in a dose 2-4 g/day to patients with streptococcal endocarditis (SE). Of them, 8, 8 and 1 patients were infected with Streptococcus viridans, Streptococcus pyogenes, Pneumococcus, respectively. Hospital trial and follow-up lasted up to 2 years. Cure without surgical correction occurred in 7 patients, valvular replacement was needed in 4 patients, 1 patient died, 4 patients developed recurrences 3 and more months after therapy with ceftriaxone. The drug was safe and well tolerated. 33% of the patients were discharged from hospital with obvious improvement after 2-3 week treatment to continue it outpatiently for 1-2 weeks.