Renal transplantation under prophylactic eculizumab in atypical hemolytic uremic syndrome with CFH/CFHR1 hybrid protein.

We report the first observation of successful kidney transplantation under pre-emptive eculizumab treatment in a 7-year-old boy with atypical hemolytic uremic syndrome (aHUS) and a known hybrid CFH/CFHR1 gene, who was dependent on plasma therapy during the 3-year dialysis period. The hybrid CFH/CFHR1 protein has an altered C3b/C3d binding, is incapable to protect cells from complement attack and is directly implicated in aHUS pathogenesis. There was no evidence of recurrence during the first 16-month follow-up period. We conclude that eculizumab alone, without plasma therapy (plasma infusion and/or plasma exchange), is sufficient to prevent recurrence of aHUS and to maintain long-term graft function.

Eculizumab for atypical hemolytic uremic syndrome recurrence in renal transplantation.

Eculizumab (anti-C5) has been sporadically reported as an efficient therapy for atypical hemolytic uremic syndrome (aHUS). However, the lack of series precludes any firm conclusion about the optimal use of anti-C5 for preventing or treating aHUS posttransplant aHUS recurrence. We thoroughly studied 22 renal transplant recipients with aHUS who received off-label therapy with anti-C5, including 12 cases, which have not been reported yet. Nine patients, all carrying a complement genetic abnormality associated with a high risk of aHUS recurrence, received prophylactic anti-C5 therapy to prevent posttransplant recurrence. Eight of them had a successful recurrence-free posttransplant course and achieved a satisfactory graft function, while the remaining patient experienced early arterial thrombosis of the graft. Thirteen renal transplant recipients were given anti-C5 for posttransplant aHUS recurrence. A complete reversal of aHUS activity was obtained in all of them. Importantly, the delay of anti-C5 initiation after the onset of the aHUS episode inversely correlated with the degree of renal function improvement. Three patients in whom anti-C5 was subsequently stopped experienced a relapse. Altogether these data suggest that long-term eculizumab is highly effective for preventing and treating posttransplant aHUS recurrence. Our study also indicates that anti-C5 should be promptly started if a recurrence occurs.

First Report of Fruit Rot of Olives Caused by Botryosphaeria dothidea in Tunisia.

During the summer of 2010, unfamiliar symptoms of fruit rot were frequently observed on different Tunisian olive (Olea europaea) cultivars. These symptoms appeared to be associated with the damage caused by the olive fruit fly (Bactrocera oleae). At first, infected olives showed a brown color and then fruits begin to depress until they become completely mummified and fall immaturely. This problem was more pronounced on table olive cultivars (Ascolana, Meski, and Picholine) in the northern Tunisian regions (Nabeul), with an infection incidence of 65%. Infected Ascolana olives were disinfected with 70% ethanol for 2 min, rinsed in sterile distilled water, and air dried. Several pieces were cut and placed on acidified (2.5 ml of a 25% [vol/vol] solution of lactic acid per liter of medium) potato dextrose agar medium (PDA). All plates were incubated at 25°C for 4 days under continuous fluorescent light. A fast-growing fungus with an abundant, aerial mycelium, which was gradually veering from white to dark gray, was constantly isolated. On the reverse side of the colonies, an olive green coloration spread to the edge and became darker from the center until the underside was completely black. Conidia produced on the PDA plate were hyaline, single or double cell, ellipsoid, with a subobtuse apex and a truncate base, and averaged 22.70 × 5.32 µm. Conidiophores were hyaline, cylindrical, smooth, branched at the base, with an average of 14 to 24 × 2 to 3 µm. Pathogenicity of an isolate was conducted by dipping 20 olives wounded by a sterilized scalpel in a conidial suspension (105 conidia/ml), covering inoculated olives with moisture filter paper, and incubating them in a polyethylene bag under darkness at 25°C. Controls however, were wounded and dipped in sterile distillated water. Seven days after the inoculation, olives showed a brown color covering half of the fruit. Later (15 days after), this browning was accentuated and several black pycnidia were observed. Forty days after inoculation, fruits were completely dried out and the kernel was already appearing. Controls, however, remained totally healthy. Koch's postulates was then verified and showed that pure cultures were obtained after reisolations from inoculated olives, whereas the controls were free of the fungus. BLAST analysis of the internal transcribed spacer region (ITS) of rDNA of one isolate showed 99% identity with the ITS sequence of Botryosphaeria dothidea (GenBank Accession No. FM955381.1). Species of the family of Botryosphaeriaceae are common pathogens causing fruit rot and dieback of many woody plants (3). Drupe rot problem caused by B. dothidea was reported on olives in Greece (4) and southern Italy (2). It was reported that the fungus invades the drupes through the wounds caused by the olive fruit fly and may even be transmitted by it (1), and recently Moral et al. (3) suggested that the olive fruit fly is essential for the initiation of the disease on the fruit. To our knowledge, this is the first report of fruit rot of olives caused by B. dothidea in Tunisia. References: (1) N. González et al. Bol. San. Veg. Plagas 32:709, 2006. (2) C. Lazzizera et al. Plant Pathol. 57:948, 2008. (3) J. Moral et al. Phytopathology 100:1340, 2010. (4) A. J. L Phillips et al. Mycopathology 159:433, 2005.

First Report of a Branch Dieback of Olive Trees in Tunisia Caused by a Phoma sp.

From 2007 to 2008, a new dieback of branches of olive trees was observed in several orchards in central and southern Tunisia. The appearance of this new syndrome coincided with warm temperatures and frequent rainfall from February to April 2007. Affected trees were observed in seven commercial orchards; disease incidence ranged from 1 to 9% and affected trees were randomly distributed in each orchard. Symptoms included abundant dead branches and wilted leaves remained attached. Distinct brown areas appeared on the bark of current-year shoots as well as on larger branches. Cankers on branches that were >2 years old were difficult to detect but were conspicuous in current-year shoots. To determine the etiology of this new syndrome, a study was carried out on samples of affected branches collected from 2007 to 2008 from different areas of the country. Unidentified species of Chaetomium and Phoma were isolated from the margins of the cankers. Koch's postulates were performed with one isolate each of a Chaetomium sp. and a Phoma sp on 2-year-old olive trees, cv. Chemlali, grown in 13-cm-diameter pots containing a sand/lime/peat mixture. Stems were inoculated by placing 10 µl of conidial suspension (106 conidia/ml) on 1-cm-long longitudinal stem wounds that had been made with a sterile scalpel. Control plants were wounded, but inoculum was replaced with sterile distilled water. Three sets of 10 plants each were wound inoculated with each of the fungi on the same day. Inoculated plants were covered with a polyethylene plastic bag to retain moisture and incubated for 2 months at 30°C with a 12-h photoperiod. After 45 days, only branches inoculated with the Phoma isolate showed brown discoloration areas at the inoculation sites. A Phoma sp. was recovered from necrotic bark from each of the 10 inoculated plants. Conidia were hyaline, unicellular, slightly ellipsoidal, and 4.8 to 6.3 × 1.8 to 2.2 µm. To confirm the identification, DNA extraction was done with hyphae collected from a 7-day-old culture on PDA after incubation at 25°C (1). Fungal primers ITS1 and ITS4 (3) were used for amplification. Purified amplicons were directly sequenced using the ITS1 and ITS4 primers for the internal transcribed spacer region of rDNA. A BLAST search of the GenBank database revealed 96% homology with Phoma sp. isolate (AJ972865.1) and 98% homology with Phoma medicaginis isolate (DQ026014.1). P. incompta has been reported as responsible for branch dieback of olive tree in Italy (2). To our knowledge, this is the first report of a canker disease of olive caused by a Phoma sp. in Tunisia. References: (1) S. R. Tendulkar et al. Biotechnol. Lett. 22:1941, 2003. (2) L. Tosi and A. Zazzerini. Petria 4:161, 1994. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Delineation of Pseudomonas savastanoi pv. savastanoi strains isolated in Tunisia by random-amplified polymorphic DNA analysis.

AIMS: To investigate the genetic diversity of Pseudomonas savastanoi pv. savastanoi strains and to look whether these strains were distributed to geographical location. METHODS AND RESULTS: Random amplification of polymorphic DNA (RAPD) was used to discriminate between 58 Tunisian strains and 21 strains from various other countries of P. savastanoi pv. savastanoi, the causal agent of olive knot disease. Isolates were separated into three groups by cluster analysis and principal coordinate analysis of RAPD fingerprint data obtained with three primers (OPR-12, OPX-7 and OPX-14). Group 1 contained isolates from the southeast of Tunisia and European strains. Group 2 comprised strains isolated from the north of Tunisia exclusively while group 3 encompassed the majority of isolates obtained from five orchards located in the centre of Tunisia. CONCLUSIONS: The results indicated that isolates of P. savastanoi pv. savastanoi were genetically distinct according to geographic regions. RAPD grouped isolates derived from the same orchard as identical. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first application of RAPD in the delineation of P. savastanoi pv. savastanoi strains.

[Antiglomerular basement disease in children: Literature review and therapeutic options]. / Syndrome de Goodpasture et maladie des anticorps anti-membrane basale chez l'enfant : revue de la littérature.

Antiglomerular basement membrane glomerulonephritis is a rare autoimmune disease characterized by rapidly progressive glomerulonephritis that may be associated with pulmonary hemorrhage (Goodpasture syndrome). The disease is caused by autoantibodies (classically IgGs) directed against the α3 subunit of type IV collagen. This is a rare disease in the adult population and extremely rare in children, with a reported cumulative annual incidence at 1/106 people/year. Among scarce reported pediatric cases (n=31), most are girls (M/F sex ratio, 1:4), and the mean age at diagnoses is 9.2±4.6 years. A medical diagnosis is an emergency and is based on the identification of specific antibodies in the serum, and pathognomonic linear fixation of IgGs along the glomerular basement membrane. Without appropriate treatment, the disease is generally fulminant, and patient and kidney survival is poor. Indeed, glomerular function strongly correlates with histological lesions. The current guidelines recommend the use of plasma exchanges and immunosuppressive drugs. For the past few years, alternative therapeutics such as specific anti-B-cell antibodies (rituximab) or specific extrarenal cleansing such as immunoadsorption have been successfully used in adults. Immunoadsorptions (IAs) can remove pathogenic IgGs from the circulation and do not require plasma infusions, contrary to plasma exchanges. In this review, we discuss the key points of antiglomerular basement membrane glomerulonephritis diagnosis and conventional or alternative therapeutics.

[Systemic infantile mastocytosis: about a case with respiratory and digestive involvement]. / Mastocytose systémique du nourrisson: à propos d'un cas avec atteinte respiratoire et digestive.

UNLABELLED: Systemic mastocytosis is rare in children and is characterized by an abnormal proliferation and infiltration of mast cells in different tissues. CASE REPORT: We report a case of systemic mastocytosis presenting cutaneous symptoms during the neonatal period. Later evolution was characterized by systemic manifestations consisting of recurrent respiratory infections with wheezing and a digestive involvement that included abdominal pain, hepatosplenomegaly and a nodular, hemorrhagic infiltrate in a low esophagus. The diagnosis was confirmed by histology and biology, notably increased histamine concentrations in blood and urines. Improvement of the respiratory and digestive symptoms was obtained with treatment by histamine H1 and H2 receptors antagonists. CONCLUSION: Respiratory manifestations and nodular infiltration of the digestive tract are rare in systemic mastocytosis. The prognosis is conditioned by complications such as malignancy and the persistence of the disease till the adult age.