The ABCB6 mutation p.Arg192Trp is a recessive mutation causing the Lan- blood type.

BACKGROUND AND OBJECTIVE: The membrane transporter ABCB6 has recently been shown to carry the high-frequency red-blood-cell (RBC) antigen Lan. All the Lan- individuals genotyped so far have inherited two recessive null mutations in ABCB6. The finding of a family with the Lan- blood type occurring in two successive generations prompted this study. METHODS: Mutations in ABCB6 were searched by Sanger sequencing of exons and flanking intronic regions. Expression analysis of the Lan antigen was carried out by serology and flow cytometry. PCR-RFLP genotyping and Western blot analysis were also applied. RESULTS: All the Lan- members of this family were homozygous for c.574C>T, p.Arg192Trp in ABCB6 while the Lan+ members were heterozygous for this missense mutation encoded by the SNP rs149202834. Homozygosity for p.Arg192Trp was associated not only with absence of the Lan antigen, but also of the ABCB6 transporter in RBC membrane. The complete absence of Lan expression resulting from p.Arg192Trp homozygosity was confirmed by the subsequent identification of five unrelated Lan- individuals who were homozygous for this mutation and who developed an anti-Lan. We also provide evidence that three other single amino acid mutations in ABCB6 (c.826C >T, p.Arg276Trp; c.85_87delTTC, p.Phe29del; c.1762G >A, p.Gly588Ser) may also define ABCB6 null alleles. CONCLUSION: p.Arg192Trp is the first ABCB6 missense mutation causing the Lan- blood type and appears to be a relatively frequent cause of this rare blood type. Like the previously reported frameshift, nonsense and essential splice-site mutations in ABCB6, this missense mutation is recessive and defines an ABCB6 null allele. Other single amino acid mutations in ABCB6 may also cause the Lan- blood type.

A new AQP1 null allele identified in a Gypsy woman who developed an anti-CO3 during her first pregnancy.

BACKGROUND AND OBJECTIVES: The Colton blood group antigens are carried by the AQP1 water channel. AQP1(-/-) individuals, also known as Colton-null since they express no Colton antigens, do not suffer any apparent clinical consequence but may develop a clinically significant alloantibody (anti-CO3) induced by transfusion or pregnancy. Identification and transfusion support of Colton-null patients are highly challenging, not only due to the extreme rarity of this phenotype, the lack of appropriate reagents in most laboratories, as well as the possibility of confusing it with the recently described CO:-1,-2,3,-4 phenotype where AQP1 is present. This study investigated a new Colton-null case and evaluated three commercially available anti-AQP1s to identify Colton-null red blood cell samples. METHODS: The Colton-null phenotype was investigated by standard serological techniques, AQP1 sequencing, immunoblot and flow cytometry analyses. RESULTS: We identified and characterized the Colton-null phenotype in a Gypsy woman who developed an anti-CO3 during her first pregnancy. After developing a simple and robust method to sequence AQP1, we showed that she was apparently homozygous for a new AQP1 null allele, AQP1 601delG, whose product is not expressed in her red blood cells. We also established the Colton specificity of three commercially available anti-AQP1s in immunoblot and/or flow cytometry analyses. CONCLUSION: This Gypsy woman represents the sixth Colton-null case characterized at the serological, genetic and biochemical levels. The validation here of new reagents and methods should facilitate the identification of Colton-null individuals.

Effect of spatial heterogeneities of water fluxes and application pattern on cadusafos fate on banana-cultivated andosols.

In tropical humid environments under intensive banana production, pesticide transfer in waters can be of particular concern due to heavy rainfall, steep slopes, and soils with high infiltration capacities. The transfer in percolation and runoff waters of the nematicide cadusafos was investigated during a three month field experiment. The spatial heterogeneity of the banana plantation was taken into account by measuring percolation fluxes both under the banana plants and in the interrows with a specially designed lysimeter device installed at 60 cm depth. At the field scale, 0.34% of the pesticide applied was transferred in percolation, 0.13% in runoff. Forty-nine percent of cadusafos losses occurred by percolation under the banana plants, 23% by interrow percolation, and 28% by runoff. Losses were highest during the three weeks following cadusafos application, and this is also when dissipation in the soil was highest (calculated half-life in the soil: 7d). After this period, losses of cadusafos were low, both in soil and waters. Under the banana plant, saturated fluxes carried most of the pesticide, despite total percolation fluxes being at least five-times higher than saturated ones. Although overall pesticide transfer in water was low (0.5% of applied), it was not negligible due to the frequency of pesticide application in these areas.

Incidence of class A extended-spectrum beta-lactamases in Champagne-Ardenne (France): a 1 year prospective study.

OBJECTIVES: To assess the frequency and diversity of extended spectrum beta-lactamases (ESBLs) in the Champagne-Ardenne region France, and to identify genetic elements associated with the bla(CTX-M) genes. METHODS: During 2004, all the non-duplicate isolates of Pseudomonas aeruginosa and Acinetobacter baumannii resistant to ceftazidime and of Enterobacteriaceae intermediate or resistant to ceftazidime and/or cefotaxime, screening samples excluded, were collected in 10 public hospitals and 3 private clinics. bla genes were sequenced and bla(CTX-M) environment characterized by PCR mapping. RESULTS: In Enterobacteriaceae (138/21 861; 0.6%), ESBLs were predominantly TEM-24 (n = 52; 37.7%) and CTX-M-15 (n = 37; 26.8%). Three new enzymes were identified, CTX-M-61 (CTX-M-1 group), TEM- and SHV-type. A. baumannii (n = 5) produced VEB-1 and P. aeruginosa (n = 2) SHV-2a. ISEcp1 was detected in 22/27 strains, disrupted in 7 of them. The IS903-like element was downstream of bla(CTX-M-14) and bla(CTX-M-16). ISCR1 was found upstream of bla(CTX-M-2) and bla(CTX-M-9), and ISCR1 and bla(CTX-M-2) were located on a sul1-type class 1 integron. In comparison with 2001-02, ESBL distribution among Enterobacteriaceae showed an increase in CTX-M-type (44.9% vs 3.7% P < 10(-7)) due to Escherichia coli CTX-M-15 and to the almost total disappearance of TEM-3 (0.9% vs 51.2%). E. coli was the most frequent species (50.0% vs 5.1% in 1998) despite a similar prevalence to that in 1998 (0.5% vs 0.2%). CONCLUSIONS: A careful detection of bla(CTX-M)-type spread to other species would help to anticipate clonal endemics such as those observed in Enterobacter aerogenes TEM-24.

Septicemia due to Yersinia enterocolitica in a hemodialyzed, iron-depleted patient receiving omeprazole and oral iron supplementation.

Septicemia occurred in a long-term hemodialysis patient on oral iron supplementation who had been treated for esophageal ulcer by omeprazole, an ulcer-healing drug. Yersinia enterocolitica serotype 0:3 was recovered from blood cultures. A raised intraintestinal pH and an increased intraluminal iron load may have been contributing factors for the enhanced proliferation and generalized infection of Y enterocolitica.

[In vitro effect of combinations of netilmicin and amikacin with fosfomycin and pefloxacin on 31 strains of heterogeneous methicillin- resistant Staphylococcus aureus]. / Activité in vitro des associations de la nétilmicine et de l'amikacine avec la fosfomycine et la péfloxacine sur trente et une souches de Staphylococcus aureus méticilline-résistantes hétérogènes.

The Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) of 31 strains of Staphylococcus aureus resistant to methicillin and gentamicin were determined towards aminoglycosides: netilmicin (N) and amikacin (A). The results of the study showed a large discordance between the technic using dilution in Mueller-Hinton broth and the technic using agar diffusion with antibiotic discs. The level of sensitivity appears to decrease by dilution. The problem of using these aminoglycosides in combination with other antibiotics as fosfomycin (F) and pefloxacin (P) was faced. In vitro, antibacterial activity of combinations [N-F], [N-P] and [A-F], [A-P], was studied by microtiter checkerboard method. The antibacterial effects of these combinations were evaluated by determination of Fractional Bactericidal Indices (FBC-indices). The combination netilmicin or amikacin with pefloxacin has an additive bactericidal effect in most cases, without discrimination between the different aminoglycosides (FBC #0.82). Netilmicin or amikacin in combination with fosfomycin are found to be additive or moderately synergistic (FBC #0.53). No occurrence of antagonism was observed.