Molecular epidemiology reveals genetic diversity among 363 isolates of the Cryptococcus neoformans and Cryptococcus gattii species complex in 61 Ivorian HIV-positive patients.

Cryptococcal meningitis is a severe opportunistic infection in HIV-infected patients. In Ivory Coast, despite the availability of antiretroviral treatment (ART), this infection is still prevalent. The study investigates the genetic diversity of 363 clinical isolates of Cryptococcus from 61 Ivorian HIV-positive patients, the occurrence of mixed infections and the in vitro antifungal susceptibility of the isolates. Serotyping was performed via LAC1 and CAP64 gene amplification. Genotyping was performed using the phage M13 core (GACA)4 and (GTG)5 primers and restriction fragment length polymorphism analysis of the URA5 gene. By PCR fingerprinting, the presence of the three serotypes were demonstrated among the 363 isolates in the population studied: A (n=318; 87.6%), AD (n=40; 11%) and B (n=4; 1.1%). Using PCR fingerprinting with primers M13 (GACA)4 and (GTG)5 , we grouped the isolates into 56 molecular subtypes. We observed a high frequency (39.3%) of mixed infections, with up to two different genotypes per sample. None of the isolates were resistant to amphotericin B. Only 0.3% and 1.1% of the isolates were resistant to fluconazole and flucytosine respectively. This study revealed the high genetic diversity among Cryptococcus isolates, the occurrence of mixed infections and a high antifungal susceptibility for the majority of Ivorian cryptococcal isolates.

[Recommendations and management in Home Blood Transfusion: Levers, brakes and prospects]. / Recommandations et prise en charge transfusionnelle en HAD : leviers, freins et perspectives.

In our roles as regional coordinator for haemovigilance and transfusion safety (RCHTS - CRHST), Correspondant for haemovigilance in a medical and surgical obstetrics/hospital home care centre (MSO/HHCC-MCO/HAD) and a doctor responsible for haemovigilance in HHC, we shared our interdisciplinary viewpoints and compared our practices in an attempt to gain a better understanding of the factors that hinder and facilitate the development of blood transfusion in HHC. Despite the revaluation of tariffs for blood transfusion in HHC (effective since March 2018), and the widespread circulation of recommendations for good practice in the transfusion of packed red blood cells in HHC (April 2018), this sector is still struggling to develop. On completing our analysis, we were able to demonstrate that while our institutional and regional work continues to be essential, the power to move things forward now lies in the hands of a national authority. In our opinion, using this power would give strong, national recognition of the vital role which HHC played at a nationwide level throughout the recent health crisis.

[Conditions for blood components return, after storage in a depot]. / Conditions de reprise des produits sanguins labiles (PSL) conservés dans un dépôt de sang.

Recent changes in the regulation of blood components depots has allowed the Etablissement français du sang (EFS) to re-introduce blood components in the stock when they come back from the health establishment. This process is the responsibility of both EFS and health establishment. So collaboration between them is necessary to set an organisation to guarantee that blood components are transported and stored at optimum conditions. To achieve this, a checking procedure for the return process of blood components and a method that tracks the conditions they are exposed to during transportation are necessary. There are many temperature sensitive indicators available to track each unit of blood component. This article presents an organisation using a follow-up by electronic temperature monitors to record the temperature during the transportation and storage processes.

Comparative typing analyses of clinical and environmental strains of the Cryptococcus neoformans/Cryptococcus gattii species complex from Ivory Coast.

PURPOSE: The aim of this study was to assess the biotope of the Cryptococcus neoformans/Cryptococcus gattii species complex from Ivory Coast, and clarify the possible epidemiological relationship between environmental and clinical strains. METHODOLOGY: Samples from Eucalyptus camaldulensis (n=136), Mangifera indica (n=13) and pigeon droppings (n=518) were collected from different sites close to the living environment of Ivorian HIV patients with cryptococcosis (n=10, 50 clinical strains). Clinical and environmental strains were characterized by molecular serotyping and genotyping [RFLP analysis of the URA5 gene, (GACA)4, (GTG)5 and M13 PCR fingerprinting] and compared.Results/Key findings. Environmental strains were recovered only from the pigeon droppings. In vitro susceptibility profiles showed that all strains were susceptible to fluconazole, flucytosine and amphotericin B. All environmental strains consisted of C. neoformans (A, AFLP1/VNI), whereas clinical strains included C. neoformans (A, AFLP1/VNI), C. neoformans x Cryptococcus deneoformans hybrids (AD, AFLP3/VNIII) and Cryptococcus deuterogattii (B, AFLP6/VGII). Two patients were co-infected with both C. neoformans and C. neoformans x C. deneoformans hybrids. We noticed a low genetic diversity among the environmental samples compared to the high diversity of the clinical samples. Some clinical strains were genetically more similar to environmental strains than to other clinical strains, including those from the same patient. CONCLUSION: These results provide new information on the ecology and epidemiology of the C. neoformans/C. gattii species complex in Ivory Coast.

Erratum to: Impact of human papillomavirus on head and neck squamous cell cancers in Gabon.

[This corrects the article DOI: 10.1186/s13027-015-0036-7.].

First case of mixed infection with Cryptococcus deuterogattii and Cryptococcus neoformans VNI in an Ivorian HIV-positive patient.

INTRODUCTION: Cryptococcal meningitis (CM) may be caused by several species of Cryptococcus. CASE PRESENTATION: We describe a fatal case of CM in a HIV-positive patient from Ivory Coast infected by Cryptococcus neoformans VNI and Cryptococcusdeuterogattii. Isolates were recovered from cerebrospinal fluid (CSF) prior to systemic antifungal treatment. Six isolates were studied (the entire culture plus five isolated colonies from it). Serotyping was performed via LAC 1 and CAP 64 gene amplification. Genotyping was performed using restriction fragment length polymorphism (RFLP) analysis of the URA5 gene, (GACA)4, (GTG)5 and M13 PCR fingerprinting. URA5-RFLP analysis identified the original culture with two different molecular type combinations. However, URA5-RFLP profiles of the five colonies isolated from the original sample revealed two different species. Four colonies were identified as C.deuterogattii and the last isolate as C.neoformans VNI. The in vitro susceptibility profile was determined using the standard method according to the CLSI M27-A3 protocol. The isolates were susceptible to the tested antifungals (fluconazole, flucytosine and amphotericin B). Treatment with fluconazole (1200 mg day-1) was initiated; however, the patient died 17 days after the onset of antifungal therapy. CONCLUSION: This is the first reported case of mixed infection with C. neoformans and C.deuterogattii in a HIV-positive patient.

[What to do in case of adverse event in recipient at the French National Blood Service?]. / Conduite à tenir en cas d'effet indésirable receveur à l'établissement français du sang.

One of the main goals of haemovigilance is to gather and analyze adverse events in recipients of blood products in order to improve blood safety. The French National Blood Service has a specific role in the management of immediate adverse events: to alert to quarantine the potentially dangerous blood products from the same donation(s), to provide blood testing for the etiologic assessment and to give transfusion advice to patients. The updating of the recipient's computer file allows a better monitoring for both immediate and delayed adverse events. Finally, the French National Blood Service's correspondent of haemovigilance is responsible for donor's inquiries, especially in cases of transfusion related to bacterial contamination, severe allergy, suspicion of transfusion acute related lung injury and viral seroconversion. The management effectiveness for adverse events requires a strong collaboration between all members of the haemovigilance network.

[Use of platelet concentrates: a regional experience]. / Utilisation des concentrés plaquettaires : expérience d'une région.

PURPOSE OF THE STUDY: The Établissement français du sang (EFS) distributes two types of platelet concentrates: using a single donor in aphaeresis platelet concentrate (SDAP), versus pooled platelet concentrates (PPC). A retrospective study performed by the Blood Derivatives Group at Observatory for Drugs, Medical Devices and Therapeutic Innovations (OMEDIT), in collaboration with EFS and haemovigilance correspondents from eight regional health care establishments, has analyzed platelet concentrates prescriptions and the position of the prescribers concerning PPC supply. MATERIAL AND METHODS: Between the 2nd and 6th June 2008, 151 platelet concentrates were supplied by ESF. Data were collected for 144 platelet concentrates and in 83 transfused patients with an average age of 50years. During this study, 33 PPC (23%) and 111 SDAP (77%) were supplied. RESULTS: With regards to the 111 SDAP, the supply of PPC was refused in 47 cases (42%), accepted in 18 cases (16%) and unknown for 46 cases (42%). A total of 51 PPC could be supplied during this period, which represented 35% of platelet concentrates prescriptions. The rate of platelets before transfusion was known for 121 platelet concentrates, the median was 32G·L(-1) for SDAP and 44G·L(-1) for PPC. CONCLUSION: More frequent PPC use, with comparable therapeutic efficacy, could be interesting in a context of increasing platelet concentrates consumption in health care establishments. Moreover, prescribers did not seem to be against the idea. An information pamphlet on platelet concentrates was drafted and distributed to prescribers in order to promote the prescription of PPC. A second assessment is planned to measure the impact of this communication.

The location and nature of general anesthetic binding sites on the active conformation of firefly luciferase; a time resolved photolabeling study.

Firefly luciferase is one of the few soluble proteins that is acted upon by a wide variety of general anesthetics and alcohols; they inhibit the ATP-driven production of light. We have used time-resolved photolabeling to locate the binding sites of alcohols during the initial light output, some 200 ms after adding ATP. The photolabel 3-azioctanol inhibited the initial light output with an IC50 of 200 µM, close to its general anesthetic potency. Photoincorporation of [(3)H]3-azioctanol into luciferase was saturable but weak. It was enhanced 200 ms after adding ATP but was negligible minutes later. Sequencing of tryptic digests by HPLC-MSMS revealed a similar conformation-dependence for photoincorporation of 3-azioctanol into Glu-313, a residue that lines the bottom of a deep cleft (vestibule) whose outer end binds luciferin. An aromatic diazirine analog of benzyl alcohol with broader side chain reactivity reported two sites. First, it photolabeled two residues in the vestibule, Ser-286 and Ile-288, both of which are implicated with Glu-313 in the conformation change accompanying activation. Second, it photolabeled two residues that contact luciferin, Ser-316 and Ser-349. Thus, time resolved photolabeling supports two mechanisms of action. First, an allosteric one, in which anesthetics bind in the vestibule displacing water molecules that are thought to be involved in light output. Second, a competitive one, in which anesthetics bind isosterically with luciferin. This work provides structural evidence that supports the competitive and allosteric actions previously characterized by kinetic studies.

[The management of post donation information: a fundamental element of blood safety]. / La gestion des informations post-don: un élément fondamental de la sécurité transfusionnelle.

Post donation information management is a fundamental axis of haemovigilance in terms of blood safety. It requires an organization ensuring a permanent reactivity, a good sensitization of French National Blood Service professionals and needs also a strong awareness of blood donors. Previous identification of stakeholders to warn during these kinds of alerts is essential to avoid the use of any blood product presenting a potential risk. The recent implementation of a consensual internal document aims to target the reinforcement of a homogeneous decision-making process, combining blood product self-sufficiency and above all recipient safety.

[Suitability of red blood cell transfusion: a multicenter study]. / Évaluation multicentrique de la pertinence des prescriptions de concentrés de globules rouges.

The purpose of this retrospective observational multicenter study was to assess appropriateness of red blood cell (RBC) transfusion, according to the French national guidelines (Agence française de sécurité sanitaire des produits de santé) published in 2002. Six hundred and thirty-nine RBC transfusions from nine institutions have been randomly selected and analysed. The data collected are issued from different specialities. Patients' characteristics, occurrences of transfusion, admission, pre-transfusion, post-transfusion and discharge haemoglobin concentrations have been collected. Two physicians (who are in charge) must evaluate the appropriateness of pre-transfusion, discharged haemoglobin concentrations, quantity and quality of transfused RBC. The mean pre-transfusion haemoglobin concentration was 7.89 ± 1.24, the median number of transfused RBC was two (extremes: 1-16), the mean discharge haemoglobin concentration was 10.14 ± 1.30 (-5 days after the end of transfusion). The pre-transfusion and discharge haemoglobin concentrations were higher if the patient presented a co-morbidity factor. Ninety-three percent of pre-transfusion and 79% of discharge haemoglobin concentrations are in accordance with the guidelines. According to the physicians, the RBC transfusions are too "precocious" when pre-transfusion haemoglobin concentration is above nine and the anaemia is asymptomatic. 50% of RBC transfusion with discharge haemoglobin concentration above 10 is not excessive. In case of acute anaemia, the pre-transfusion and discharge haemoglobin concentrations are higher and RBC transfusion excessive. In this study, the trigger haemoglobin concentration is "restrictive", but the target haemoglobin concentration is "liberal" with a high-discharge haemoglobin concentration. Inappropriate RBC transfusions are mainly due to over-transfusion.