Complement activation in thrombotic thrombocytopenic purpura.

BACKGROUND: Ultra-large von Willebrand factor and deficiency of its cleaving protease are important factors in the events leading to thrombotic microangiopathy; however, the mechanisms involved are only partly understood. Whereas pathological activation of the alternative complement pathway is linked to atypical hemolytic uremic syndrome, the role of complement activation in thrombotic thrombocytopenic purpura (TTP) is unknown. The aim of this study was to investigate whether signs of complement activation are characteristic of TTP. PATIENTS AND METHODS: Twenty-three patients with TTP (18 women, median age 38 years) and 17 healthy controls (13 women, median age 38 years) were included. Complement parameters (C3, Factors H, I, B and total alternative pathway activity) together with complement activation fragments (C3a) or complexes (C1rs-INH, C3bBbP, sC5b9) were measured by ELISA or RID. ADAMTS13 activity and anti-ADAMTS13 inhibitory antibodies were measured by the VWF-FRET73 assay. RESULTS: Increased levels of C3a, and SC5b9 were observed in TTP during acute episodes, as compared with healthy controls. Decreased complement C3 levels indicative of complement consumption occurred in 15% of acute TTP patients. Significant decrease of complement activation products C3a and SC5b9 was observed during plasma exchange (PEX). The sustained presence of anti-ADAMTS13 inhibitory antibodies in complete remission was associated with increased complement activation. CONCLUSION: These data document in an observational study the presence of complement activation in TTP. Further investigation is needed to determine its potential pathogenetic significance.

Diagnosis and classification of hemolytic uremic syndrome: the Hungarian experience.

BACKGROUND: Hemolytic uremic syndrome (HUS) is a rare disease with various etiologies, making the identification of the specific forms and appropriate treatment difficult. Therefore, clinical and laboratory data from these patients need to be analyzed in national and international registries. Herein we have described 47 Hungarian HUS patients with detailed laboratory and clinical data obtained between 2008 and 2010. METHODS: Blood samples and clinical data of 47 patients with HUS diagnosed according to characteristic clinical signs were submitted for diagnostic evaluation, including complement protein and genetic analysis, measurement of ADAMTS13 activity and antibody analysis against O157LPS and factor H. RESULTS: There were 8 patients with typical diarrhea-positive HUS; 13 with atypical HUS (aHUS) and 26 with secondary HUS/thrombotic thrombocytopenic purpura group characterized by signs of complement consumption and decreased ADAMTS13 activity. Thus, decreased total alternative pathway activity is a promising diagnostic parameter with good sensitivity for aHUS. CONCLUSIONS: These observations highlight the requirement for multiple diagnostic tests together with clinical data to identify the specific cause of HUS. Because the long-term prognosis of aHUS, eg, graft survival after renal transplantation, may vary according to the molecular etiology, it is important for all affected patients to undergo a detailed molecular diagnosis of the disease. There is a clear clinical need for the development and application of novel assay in this field to allow more rapid efficient diagnosis of patients who undergo a first episode of HUS.

Synthesis and bioimmunological efficiency of poly(2-oxazolines) containing a free amino group.

Novel amphiphilic copolymers on the basis of 2-oxazolines containing a free amino group were prepared. The copolymers were synthesized by the living cationic polymerization of 2-ethyl-2-oxazoline (ETOX) and 2-(4-aminophenyl)-2-oxazoline (APOX). The main goal of this work was the synthesis of water soluble polymer material with the defined number of functional groups necessary for the attachment of proteins and polysaccharides. A high concentration of free amino groups allows immobilization of various biosubstances, e.g. drugs, proteins or polysaccharides. Thermal properties have been studied with respect to the composition of the copolymers. Cytotoxicity and the bioimmunological efficiency of the selected copolymer were studied.

Transvesical placement of one limb of an aortobifemoral bypass graft as a complication of aortic bypass surgery.

We report about a rare complication after aortobifemoral bypass surgery. In the present case the left limb of an aortobifemoral bypass graft was placed straight through the cavity of the urinary bladder. The diagnosis was found and ascertained by colour duplex ultrasound examination, cystoskopy and angiography. In consequence, we successfully performed open bladder surgery to replace the misplaced graft.

Analysis of lower extremity embolic material after total knee arthroplasty in a canine model.

Emboli assumed to consist of fat, marrow, cement, and bone are generated during total knee arthroplasty. This study collected and identified the constituents of the embolic debris and assessed the effects of intramedullary cutting guides in a canine model. During cemented knee arthroplasties with or without the use of intramedullary guides, all embolic debris was collected in a single lobe of lung. In dogs using intramedullary guides, transesophageal echocardiography revealed a sustained embolic shower, and histologically marked amounts of emboli obstructed the pulmonary vasculature. The emboli were composed of fat cells and hematopoietic cells (marrow elements) interspersed with free fat globules. Marrow elements are the primary constituents of the emboli generated during total knee arthroplasty. Intramedullary guides increased debris generation.

Analysis of hopanoids in bacteria involved in food technology and food contamination.

Hopanoids are pentacyclic triterpenoids which are believed to act as reinforcers of membranes in certain prokaryotic microorganisms. A rapid and sensitive method for their screening in bacteria was elaborated, involving extraction of nonsaponifiable lipids and the analysis by gas chromatography-mass spectrometry, selectively monitoring the ion of m/z = 191. Using the method, hopanoids were detected in strains of Acetobacter pasteurianus, but were found to be absent in lactic acid bacteria (Lactobacillus spp., Lactococcus spp.) and in food-contaminating bacteria (Salmonella spp., Listeria spp., Yersinia spp. and others).

Inhibition of sterol 4-demethylation in Candida albicans by 6-amino-2-n-pentylthiobenzothiazole, a novel mechanism of action for an antifungal agent.

The effects of 6-amino-2-n-pentylthiobenzothiazole (APB), a new antifungal agent, on ergosterol biosynthesis in Candida albicans and Saccharomyces cerevisiae were studied, using [14C]acetate incorporation. In C. albicans, the inhibition of growth was accompanied by a marked inhibition of acetate incorporation in 4-desmethylsterols, with a significant portion of the radiolabel being incorporated in 4,4-dimethylsterols, lanosterol, and 4,4-dimethylzymosterol and minor amounts being incorporated in 4-methylsterols and squalene. The data are interpreted as evidence of a block of the ergosterol biosynthesis pathway at the level of 4-demethylation of 4,4-dimethylzymosterol, with partial inhibition of lanosterol 14-dimethylation and squalene epoxidation also being possible. In 6-amino-2-n-pentylthiobenzothiazole-treated S. cerevisiae, a significant amount of the radiolabel was incorporated also in 4-methylsterols, 4-methylzymosterol, and 4-methylfecosterol, indicating that in this microorganism there are different sensitivities of the two 4-demethylations and that the pathway is blocked at the level of 4-demethylation of 4-methylsterols.

Egg dispersion in codling moth: Influence of egg extract and of its fatty acid constituents.

Codling moth females (Cydia pomonella, Lepidoptera: Tortricidae) (CM) usually lay single eggs and have a tendency to disperse. In a first experiment we observed that single females exposed to 20 apples distribute their eggs regularly among apples, suggesting a dispersive oviposition behavior. In a dual-choice situation, isolated females avoided oviposition on areas of cardboard treated with a methylene dichloride egg extract at the dose of ca. 1.0 egg equivalent/cm(2). A strong avoidance was obtained in response to a 20-fold dose, which was accompanied by a significant reduction of total oviposition. Seven major compounds found by GC analyses in the methylene dichloride extract of 2 to 3-day-old eggs were saturated or unsaturated C14-C18 straight-chain fatty acids: myristic acid, palmitic acid, palmitoleic acid, stearic acid, oleic acid, linoleic acid, and linolenic acid. This was confirmed by GC-MS analysis of an extract made by ethyl ether. Smaller amounts of three methyl esters were also identified as methyl myristate, methyl palmitate, and methyl stearate. A blend of the seven fatty acids (FA) mimicked rather well the avoidance provoked by the extract, and this avoidance was confirmed by choice between treated and untreated fruits by single females. The treatment of apples with the 7FA mixture induced an aggregative distribution of the eggs among apples. We also found that the amounts of fatty acids harvested in the extract depend on the egg age. Amounts of fatty acids increased until eggs were 4 days old and then decreased before hatching. In this paper we discuss the possible role of simple molecules, such as fatty acids and their esters, as semiochemicals indicative of juvenile tissues.

Integrating disposable or planned replacement lenses into contact lens practice.

Disposable and planned replacement lenses have become increasingly popular options for contact lens wearers. Optometrists must devise management strategies to avoid the many problems that have emerged from the use of these lens modalities: inappropriate patient selection, reduced frequency of eye examination, oversupply of lenses, improper policies for the release of contact lens prescriptions, and inadequate documentation of service contracts. Suggestions are offered to avoid these common management problems.

Stress-induced changes of glucocorticoid receptor in rat liver.

The effect of corticosterone injection and of acute and repeated stress on rat liver cytosol glucocorticoid receptor was studied to ascertain whether corticosterone-induced glucocorticoid receptor (GR) regulation also takes place in intact animals as it does in adrenalectomized ones. Adult male rats were exposed to six different stressors (swimming, 10 mg/kg histamine i.p., 500 mU/kg vasopressin s.c., heat, immobilization and cold) acutely or three times daily for 18 days (repeated stress). Each of the stressors applied acutely provoked a pronounced increase of plasma corticosterone with subsequent induction of hepatic tyrosine aminotransferase activity. Depletion of cytosol receptor was however only noticed after swimming and histamine injection. On the other hand, sustained hypersecretion of corticosterone evoked by repeated stress significantly reduced the number of GR in rat liver cytosol without any change in Kd. It is concluded that in the presence of intact adrenal glands cytosol receptors are more resistant to corticosterone-induced depletion than in their absence. Further, repeated stress causes down-regulation of GR in the liver, most probably by sustained corticosterone secretion, yet the effect of other stress factors cannot be excluded.

Deleterious cardiac effects of captopril during acute myocardial ischaemia in the dog.

OBJECTIVE: The aim was to evaluate the effects of the angiotensin converting enzyme inhibitor captopril on acute myocardial ischaemia. METHODS: Seventeen anaesthetised open chest dogs were randomised to 3 minute angioplasty balloon occlusions of the left circumflex coronary artery before and after intravenous infusion of captopril (n = 8) or placebo (n = 9). RESULTS: There was apparent worsening of ischaemia during balloon inflation after captopril infusion, when compared with control inflation, as suggested by further ST segment elevation of 1.8 (SD 1.8) mm, p less than 0.03, and by further lowering of regional myocardial pH [-0.05(0.05), p = 0.06], and peak positive and peak negative dP/dt [-439(337)mm Hg.s-1, p less than 0.008; -470(316) mm Hg.s-1, p less than 0.004, respectively]. The increase in ischaemia occurred despite reduced double product after captopril administration. Regional myocardial blood flow in the ischaemic artery distribution was lower during post captopril balloon occlusion [-0.1(0.06) ml.min-1.g-1, p less than 0.005] than during control balloon inflation, while coronary vascular resistance increased by 161(172)% (range 45 to 497%, p less than 0.04). There were no significant differences in ST segments, pH, haemodynamic variables, or blood flow during balloon inflations before and after saline infusion. CONCLUSIONS: Despite lower myocardial metabolic demands, acute intravenous administration of captopril was associated with increased ischaemia during transient coronary artery occlusion.

Floral volatiles ofTanacetum vulgare L. attractive toLobesia botrana den. et schiff. females.

The European grapevine moth (EGVM),Lobesia botrana, is a major pest of grapes in Europe. Females are attracted to a nonhost plant: tansy (Tanacetum vulgare L.), which is a common weed in Slovakian vineyards. A steam distillate extract of tansy flowers was analyzed by means of a GC-EAG technique to screen constituents detected by the olfactory receptors of EGVM females. From more than 200 GC peaks, nine peaks corresponding to monoterpenoids released an EAG response in more than 70% of the females (N=15):p-cymene,d-limonene,α-thujene,α-thujone,ß-thujone, thujyl alcohol, terpinene-4-ol, (Z)-verbenol, and piperitone. The steam distillate of tansy as well as a synthetic blend of identified compounds released consistent attraction in a field cage. The use of nonhost plants and host plant odors in integrated pest management is discussed.

[Diagnosis and treatment of thromboembolic diseases of the right heart]. / A jobbszívfél thromboemboliás betegségeinek diagnózisa és kezelése.

Right heart thrombosis (RHT) was found by 2D-echocardiography in 8 cases. Clinical suspicion of RHT could be documented in only 3 patients, while in the other 5 cases syncope, low output syndrome, essential pulmonary hypertension, cerebral embolism or congestive heart failure was the clinical diagnosis on first presentation. Out of the 4 cases of mobile RHT of extracardiac origin 1 patient had an emergency operation, 2 patients died shortly after the 2D-echo diagnosis before treatment could have been started and 1 patient improved on anticoagulant treatment. RHT of intracardiac origin was due to a central line or a ventriculoatrial shunt in 3 cases and no source could be found in 1 patient. Complete recovery was achieved in 2 cases by medical, in one case by surgical management and in 1 patient medical and surgical treatment resulted in clinical improvement. In conclusion authors 1. consider 2D echocardiography necessary in the clinical setting of acute or chronic pulmonary embolism or "primary" pulmonary hypertension and 2. they recommend emergency operation in case of mobile large RHT detected by 2D-echocardiography.

Determination of acrylamide in sugar by capillary GLC with alkali flame-ionization detection.

A method for the determination of acrylamide traces as a residue of anti-incrustation agents in sugar was developed. Acrylamide was extracted into ethyl acetate after derivatization by bromination to 2,3-dibromopropionamide (2,3-DBPA). The extract was cleaned up on a silica gel column and analysed by gas-liquid chromatography with an alkali flame-ionization detector (AFID). A glass capillary column with OV-1 stationary phase was used for this analysis. The recovery of the method determined in model experiments was 70.0% and 77.1% at 100 micrograms kg-1 and 20 micrograms kg-1, respectively. The limit of detection of acrylamide in sugar is 1-10 micrograms kg-1. Parameters of the described method are compared with other published methods.

Acute porphyria in the emergency department.

Patients with acute hepatic porphyria present with abdominal pain and neurologic abnormalities. Although the disease is uncommon, the emergency physician will occasionally encounter a patient with porphyria. The relevant pathophysiology of acute hepatic porphyria and the treatment of the patient with acute hepatic porphyria are reviewed.

Analysis of triazine herbicides residues in butter and pasteurized milk.

A method has been developed for the determination of atrazine, cyanazine, prometryn, simazine, and terbutryn residues in butter. The residues were extracted from the matrix with a mixture of petroleum ether/methanol (3 + 1), and from the separated water-methanol phase extraction was carried out with chloroform. The extract was cleaned up on an alumina column. Capillary glass liquid chromatography using a 15 m x 0.32 mm glass capillary column coated with OV-1 and an alcali flame ionization detector were employed for the analysis of the residues. The analyses were evaluated by the internal standard method, using metribuzin as the internal standard. The recovery of the method was 68.7%-79.8% for the individual herbicides under study at the fortification level of 0.1 mg.kg-1 and 79.2%-91.9% at the fortification level of 0.02 mg.kg-1. The determination limit of the method was 0.005 mg.kg-1. When centrifuging full milk, residues of triazines were partitioned between the water and fat phases, whereby 17%-82% of the residues were transferred to the milk fat. Samples of commercial butter were analysed and found to contain 0.005-0.023 mg.kg-1 atrazine.