Harmonization of strategies for the validation of quantitative analytical procedures. A SFSTP proposal--part II

As reported in a previous paper [1], the main objective of the new commission of the Société Française des Sciences et Techniques Pharmaceutiques (SFSTP) was the harmonisation of approaches for the validation of quantitative analytical procedures. In a series of meetings, members of this Commission have first tried to review the objectives of analytical methods and the objectives of validation methods and to recommend the use of two-sided β-expectation tolerance intervals for total error of validation samples (accuracy profile) in the acceptance/rejection of analytical method in validation phase.     

Anatomie morphologique des nerfs crâniens dans leur portion cisternale (du III au XII)

One hundred brains (first injected in cerebral arteries and veins with latex neoprene or India ink and studied under optic magnification) illustrate this anatomic chapter concerning the microsurgical anatomy of the cisternal segment, the neurovascular relationships, and the blood supply of the IIIrd to the XIIth cranial nerves.     

Reverse Iontophoresis of Amino Acids: Identification and Separation of Stratum Corneum and Subdermal Sources <Emphasis Type="BoldItalic">In Vitro</Emphasis>

Purpose  

To differentiate the stratum corneum (SC) and subdermal sources of amino acids (AAs) extracted by reverse iontophoresis.     

Anatomical basis of an original pedal approach to the great saphenous vein for surgery

Varicose vein repair often necessitates a distal approach to the great saphenous vein (GSV). The classic method involves a medial pre-malleolar approach. We propose a more distal approach via the medial surface of the foot where there is a cutaneous landmark, which can be used to locate the GSV in a simple, reliable and reproducible fashion. In 20 cadaveric feet, we dissected out the GSV exploiting the above-mentioned cutaneous landmark. We then extended the dissection up as far as the medial pre-malleolar region (from where the GSV is classically approached) in order to be able to compare the diameter of the vein at the two different points (in the foot and in the pre-malleolar region). We also compared the cutaneous landmark with ultrasonographic location of the GSV in 22 feet of healthy subjects. Both dissections and ultrasound examinations demonstrated the reliability of the cutaneous landmark. Moreover, no significant difference was detected in the diameter of the GSV at the two different approach points. This original distal approach to the GSV requires only minor modifications of current practice for varicose veins surgical treatment.     

Dialyzer fiber bundle volume during hemodialysis using large-surface dialyzers

Our aim was to measure and compare fiber bundle volume (FBV) of three large-surface dialyzers during chronic hemodialysis; FBV was evaluated in patients without heparin (n = 6), with tight heparinization (n = 6), and with large doses of heparin (n = 6). Each patient was treated consecutively with three dialyzers: Nephral ST500, Tricea210, and Optiflux200NR. FBV was measured hourly by ultrasound dilution in 54 sessions (n = 270). For all patients (without heparin, tight heparin, and large heparin doses) FBV did not vary significantly from 0 to 4 hours for the three types of dialyzers. There was no significant difference in aspect of filters/tubings at the end of dialysis and the presence of clots in the circuit assessed hourly. In addition, 6 other patients on hemodialysis without heparin were treated with Nephral ST500 alternatively with and without saline flushes; thus, 12 more sessions were monitored hourly for FBV (n = 60). There was no change in FBV from 0 to 4 hours between the two approaches, and the appearance of dialyzers/tubings was similar. We conclude that the FBV of large-surface dialyzers is well maintained during 4-hour chronic high-flux hemodialysis and that the loss of dialyzer surface does not explain the difference between prescribed and delivered dialysis dose in such circumstances. The membrane AN69ST does not appear more thrombogenic nor require saline flushes for heparin-free hemodialysis.     

PPV long-distance movement is occasionally permitted in resistant apricot hosts

The interactions between Plum pox virus (PPV), a member of the Potyvirus genus, and Prunus host plants are, up to now, poorly understood. In the current paper, fluorescence stereomicroscopy, in situ hybridisation and immunogold detection were performed in order to evaluate the virus transport and cellular distribution. The behavior of PPV in several susceptible (cv. ‘Moniqui’ and ‘Screara’) and resistant apricot genotypes (cv. ‘Harlayne’, ‘Henderson’, ‘Harcot’, ‘Goldrich’, ‘Stella’ and ‘Stark Early Orange’) were compared. Viral RNA was detected by in situ hybridisation in stem tissues close to the inoculation point, irrespective of the resistance status of the variety. Systemic infection was evidenced by virus immunodetection and by fluorescence detection of a GFP-tagged PPV in distant leaf sections. The signal obtained by in situ hybridisation colocalised with the fluorescence produced by GFP-tagged PPV in the same plant material but did not colocalise with the signal obtained by immunostaining. Intensity of the PPV infection in susceptible apricot cultivars varied depending on genotypes. The behavior of PPV in systemic leaves was clearly distinct between susceptible and resistant cultivars. While PPV was spreading widely around the major and minor veins in susceptible leaves, in the resistant apricot genotypes it was restricted to isolated spots consisting of few cells embedded in the mesophyll tissue. In summary, differences in the ability of PPV to systemically infect susceptible and resistant apricot cultivars were evident but nevertheless, long-distance transport of PPV occured in resistant apricot scions.     

Geosmithia argillacea: an Emerging Pathogen in Patients with Cystic Fibrosis

We report eight cases of airway colonization by Geosmithia argillacea in patients with cystic fibrosis. This filamentous fungus, resembling members of the genera Penicillium and Paecilomyces, was identified by molecular analysis. All patients carried a mutation on each CFTR (cystic fibrosis transmembrane conductance regulator) allele, with at least one copy of the F508del mutation. The first isolation of this fungus occurred from F508del-homozygous patients at a younger age than in F508del-heterozygous patients. Before recovery of G. argillacea, all patients were treated with itraconazole; two of them had also received voriconazole for an Aspergillus fumigatus infection. However, antifungal susceptibility patterns showed high MICs of voriconazole for all isolates, and high MICs of amphotericin B and itraconazole for the majority of them, but mostly low minimum effective concentrations (MECs) of caspofungin. The appearance and persistence of G. argillacea in the airways were not associated with exacerbation of the disease. However, the clinical implications of G. argillacea, particularly in immunocompromised patients, remain a concern, particularly given recent observations suggesting that this fungus may also cause disseminated infections.With a frequency of about 1/2,500 births in France, cystic fibrosis (CF) is the most common genetic inherited disease in the European Caucasian population (3). The disease is caused by mutations in the CFTR (cystic fibrosis transmembrane conductance regulator) gene, which encodes a chloride channel in the plasma membranes of various epithelial cell types. Several organs are affected, but the prognosis of CF essentially depends on the severity of lesions in the lungs (4). The defect in the chloride channel leads to thickening of the bronchial mucus, facilitating the entrapment of the inhaled bacteria and fungal conidia and providing a suitable environment for the growth of microorganisms. Nevertheless, airway colonization by filamentous fungi in CF is rarely found in young children and usually follows episodes of bacterial infection (mainly due to Staphylococcus aureus or Pseudomonas aeruginosa). Thus, previous bronchoalveolar epithelial lesions related to these infections may be required for the establishment of fungi in the respiratory tract. The clinical significance of isolating filamentous fungi from respiratory secretions remains a matter of debate. Recent studies, however, clearly show that they cause increased morbidity and an increased number of hospital admissions (1).Aspergillus fumigatus, Scedosporium apiospermum, and Exophiala dermatitidis are the most common clinically relevant fungi and are usually responsible for chronic airway colonization (13). Other filamentous fungi that are frequently, but only transiently, present in respiratory secretions include Paecilomyces variotii and some species belonging to the genus Alternaria, Cladosporium, or Penicillium. Some thermophilic filamentous fungi have also been described in humans, almost exclusively in CF patients. Indeed, we have previously described cases of chronic colonization by Penicillium emersonii (the anamorph state of Talaromyces emersonii) (6) and Acrophialophora fusispora (7) in CF patients.Here we report colonization by Geosmithia argillacea in eight CF patients attending three different French hospitals (Angers, Giens, and Rouen) between 1999 and 2009. All the isolates were identified by sequencing the ribosomal DNA (rDNA) genes. Additionally, the in vitro antifungal susceptibilities of the isolates were compared to the evolution of airway colonization during antifungal treatment.