Founder effect is responsible for the p.Leu131Phe heparin-binding-site antithrombin mutation common in Hungary: phenotype analysis in a large cohort.

BACKGROUND: Antithrombin (AT) is a key regulator of the coagulation. In type II deficiency, the heparin-binding-site defect (type II HBS) is considered to be relatively low thrombosis risk. OBJECTIVES: Our aims were to search for SERPINC1 mutation(s) and to describe the clinical and laboratory phenotype of a large number of AT Budapest3 (ATBp3, p.Leu131Phe) carriers and confirm the presence of a founder effect. PATIENTS/METHODS: AT-deficient patients were recruited and carriers of ATBp3, n = 102 (63 families) were selected. To investigate the founder effect, eight intragenic single nucleotide polymorphisms, a 5'-length dimorphism, and five microsatellite markers were detected. Clinical and laboratory data of the patients were collected and analyzed. RESULTS: In AT deficiency, 16 different causative mutations were found, and the great majority of patients were of type II HBS subtype. Most of them (n = 102/118, 86.5%) carried the ATBp3 mutation. The ATBp3 mutant allele was associated with one single haplotype, while different haplotypes were detected in the case of normal allele. The anti-factor Xa-based AT activity assay that we used could detect all ATBp3 patients with high sensitivity in our cohort. ATBp3 homozygosity (n = 26) was associated with thrombosis at a young age and conferred a high thrombotic risk. Half of the heterozygotes (n = 41/76, 53.9%) also had venous and/or arterial thrombosis, and pregnancy complications were also recorded. CONCLUSION: In Hungary, the founder mutation, ATBp3, is the most common AT deficiency. Our study is the first in which the clinical characterization of ATBp3 mutation was executed in a large population.

Alloantibody developed in a factor XIII A subunit deficient patient during substitution therapy; characterization of the antibody.

INTRODUCTION: In factor XIII A subunit (FXIIIA) deficiency, the development of alloantibodies is extremely rare. Only four reports have been published and the antibodies were not characterized. AIM: The aim of this study was to describe the clinical course and the laboratory diagnosis of a FXIII-A deficient patient who developed alloantibodies. METHODS: FXIII activity was assessed with an ammonia release assay. FXIII-A, FXIII B subunit (FXIII-B) and the complex plasma FXIII (FXIII-A2 B2 ) antigens were determined by ELISA. The causative mutation was detected by fluorescent DNA sequencing. The binding of alloantibody to FXIII-A2 and FXIII-A2 B2 was studied by surface plasmon resonance. The cleavage of FXIII-A by thrombin and Ca2+ -induced activation of thrombin-cleaved FXIII were followed by western blotting and activity measurement, respectively. RESULTS: FXIII activity, FXIII-A2 B2 and FXIII-A antigens were below the limit of detection in the patient's plasma. The severe FXIII-A deficiency was due to a novel homozygous mutation resulting in early stop codon (c.127C>T, p.Gln42STOP). The alloantibody bound to FXIII-A2 and FXIII-A2 B2 with equally high affinity (Kd ~10-8 ). It accelerated the elimination of administered FXIII concentrate from the circulation, interfered with thrombin and Ca2+ -induced activation and inhibited FXIII activity. Attempts to eliminate the alloantibody resulted only in transient improvement. Patient developed intracerebral haemorrhage after a minor trauma and died in spite of aggressive replacement therapy with FXIII concentrate. CONCLUSION: The anti-FXIII-A alloantibody caused an unmanageable bleeding complication. The antibody was of combined subtype which accelerated the elimination of FXIII and exerted a multiple inhibitory effect on FXIII activation/activity.

Factor XIII deficiency: complete phenotypic characterization of two cases with novel causative mutations.

Coagulation factor XIII (FXIII) exists as heterotetramer (FXIII-A2B2) in the plasma and as dimer (FXIII-A2) in cells. Activated FXIII mechanically stabilizes fibrin and protects it from fibrinolysis by cross-linking fibrin chains and α2-plasmin inhibitor to fibrin. FXIII is essential to maintaining haemostasis, and its deficiency causes severe bleeding diathesis. Due to improper laboratory practices, FXIII deficiency is considered the most under-diagnosed bleeding disorder. The aim of this study was to demonstrate in two cases how FXIII deficiency is properly diagnosed and classified, and to compare results of laboratory analysis and clinical symptoms. FXIII activity from plasma and platelets was measured by a modified ammonia release assay, while FXIII-A2B2, FXIII-A and FXIII-B antigens were determined by ELISA. The exon-intron boundaries and the promoter region of F13A1 gene were amplified by PCR and the amplified products were analysed by direct fluorescent sequencing. FXIII-A mRNA in platelets was determined by RT-qPCR. Two children with severe bleeding symptoms were investigated. In both cases FXIII activity and FXIII-A antigen were undetectable in the plasma and platelet lysate. In the plasma no FXIII-A2B2 antigen was found, while FXIII-B antigen was >30% in both cases. Proband1 was a compound heterozygote possessing a known missense mutation (c.980G>A, p.Arg326Gln) and a novel splice-site mutation (c.1112+2T>C). Proband2 was homozygote for a novel single nucleotide deletion (c.212delA) leading to early stop codon. The discovered mutations explain the severity of clinical symptoms and the laboratory data. Methods precise in the low activity/antigen range are required to draw valid conclusion on phenotype-genotype relationship.

First Report of Oidium longipes as the Causal Agent of Petunia Powdery Mildew in the United Kingdom.

In autumn 2009, during a survey of powdery mildews of solanaceous plants in the United Kingdom, petunia (Petunia × hybrida) plants showing typical symptoms of powdery mildew infections were repeatedly collected in East Malling, Rochester, and Sandringham, UK. Leaves, stems, and petals of the collected plants, grown as outdoor ornamentals, were covered by dense, sporulating, white mycelium. Conidia were ellipsoid-cylindrical, measured 20 to 30 × 10 to 15 µm, and were produced in chains. Germ tubes arose from the ends of conidia and terminated in simple, unlobed apices. Some of the conidiophores were extremely long, up to 250 µm, because the second or third cell, or sometimes the foot cell, was up to 105 to 170 µm long. Other conidiophores were shorter, with no exceptionally long cells, but all of them exhibited a few characteristics in common: their width increased from base to top, sometimes enlarging considerably at a particular point of the foot cell, and basal septa were usually located 7 to 30 µm from the point of branching. Hyphal appressoria were nipple shaped. The teleomorph stage was not found. On the basis of these characteristics, the fungus was identified as Oidium longipes, a recently described (4) and little known pathogen of petunia and other solanaceous plants (1,3). To support the identification of this fungus, DNA was extracted from conidia collected with sterile brushes from single leaves collected in Sandringham, East Malling, and Rochester with a Qiagen DNeasy Plant Kit (Qiagen, Hilden, Germany), and the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA was amplified and determined as described in Jankovics et al. (2). The three identical ITS sequences, deposited in GenBank under Accession Nos. HM156495, HM156496, and HM156497, were identical to several ITS sequences of O. longipes, such as AF250777, EU327324, and EU327325. This has also supported that the disease was caused by this species. Herbarium specimens were deposited under the Accession Nos. HAL 2373F, HAL 2374F, and HAL 2375F at the Herbarium of Martin Luther University, Halle, Germany. To our knowledge, this is the first report of O. longipes in the UK. References: (1) A. Bolay. Cryptogam. Helv. 20:1, 2005. (2) T. Jankovics et al. Phytopathology 98:529, 2008. (3) L. Kiss et al. Plant Disease 92:818, 2008. (4) M. E. Noordeloos and W. M. Loerakker. Persoonia 14:51, 1989.

The involvement of blood coagulation factor XIII in fibrinolysis and thrombosis.

It has been known for a long time that blood coagulation factor XIII (FXIII) is essential for maintaining haemostasis, its deficiency leads to severe bleeding complication. Biochemical studies have revealed that FXIII is a key regulator of fibrinolysis and, in addition to its role in haemostasis, it has also been implicated in the pathology of arterial and venous thrombosis. Most recently, the polymorphisms in the FXIII subunit genes and their influence on the risk of thrombotic diseases have stirred a lot of interest. This review, besides including the basic biochemistry of FXIII, mainly concentrates on the biochemical and clinical aspects of the involvement of FXIII in fibrinolysis and thrombosis. Biochemical aspects: Basics on the structure and activation of plasma and cellular FXIII. The enzymological features of activated FXIII and its main substrates. The interaction of FXIIIa with fibrinogen/fibrin and with components of the fibrinolytic system. The impact of cross-linked fibrin clot formation on the fibrinolytic processes. The down-regulation of FXIIIa within the fibrin clot. FXIII polymorphisms and their biochemical consequences. Clinical Aspects: FXIII level and the risk of arterial thrombosis (coronary artery disease, peripheral artery disease, ischemic stroke). The effect of FXIII subunit polymorphisms on the risk of arterial thrombotic diseases. The interplay between FXIII polymorphisms and other factors influencing the risk of arterial thrombosis. FXIII and venous thromboembolism.

Metabolic risk factors in patients with renal stones in KwaZulu Natal: an inter-racial study (Asian and Whites).

OBJECTIVE: To identify any differences between Whites and Indians in KwaZulu Natal province, South Africa, in the metabolic risk factors which predispose them to urinary stone formation. PATIENTS AND METHODS: Urinary stone disease is often a manifestation of an underlying metabolic disorder in most patients. Intrinsic and extrinsic factors affect the susceptibility of an individual to develop urinary stones. Although South African-born Indians and Whites in KwaZulu Natal share some of the same extrinsic factors, diet and genetic factors differ between the groups. In a study from April 1999 until April 2001, 140 patients were included who had a radiological diagnosis of renal calculi; they were evaluated metabolically using previously recommended methods. RESULTS: All the patients had at least one identifiable metabolic risk factor; the prevalence of the common metabolic risk factors was similar in the two groups. The prevalence of complete renal tubular acidosis (type 1) was significantly higher in the Indian patients. The most common metabolic abnormalities were hypomagnesuria and hypocitraturia, followed by low urinary volume. Hypercalciuria was not significant in this population. While Indians had lower urine volumes than Whites, Whites had significantly higher urinary calcium excretion than Indians. CONCLUSION: There were a few variations in the metabolic risk factors between Indians and Whites, and the differences could be attributed to genetic or dietary habits. The high incidence of renal tubular acidosis in Indian patients could explain the higher prevalence of urinary stone disease in this group than in other racial groups.