Differences selleck may exist among various species with regard to the requirement for ftsQ/divIB under laboratory conditions. The lon gene was identified, using SCOTS, in the livers of ducks infected with Riemerella anatipestifer (Zhou et al., 2009). The Lon protein is involved mainly in the quantitative regulation of cellular proteins; the strain that lacked the lon gene showed impaired replication in the host cell and exhibited a very sensitive phenotype to hydrogen peroxide and acidic environments

(Tsilibaris et al., 2006). Meanwhile, the Lon protein has been associated with bacterial pathogenesis; it has been demonstrated that the Brucella abortus and Salmonella typhi lon homologue is required for wild-type virulence during the initial stage of infection in mice (Robertson et al., 2000; Takaya et al., 2003). Baltes and Gerlach identified the tufA gene of Actinobacillus pleuropneumoniae in necrotic porcine tissue using SCOTS (Baltes & Gerlach, 2004). Elongation factor Tu (EF-Tu) is encoded by tuf genes and carries aminoacyl-tRNA Dasatinib to the ribosome during protein synthesis. The tuf mutation caused the ribosome to pause following the action of RNA polymerase and exposed unshielded nascent message to RNase E cleavage (Hammarlof & Hughes,

2008). In addition, scs-L7 and scs-L20, which encode peptide chain ID-8 release factor 1 and HemK protein, were identified by SCOTS analysis. In E. coli, the genes were present in the hemA-prfA-hemK

operon; PrfA belongs to the cAMP receptor protein/fumarate nitrate reductase regulator family of bacterial transcription factors, and HemK plays a role in the termination of translation (Dincbas-Renqvist et al., 2000). A hemK knockout strain of E. coli not only suffered severe growth defects, but also showed a global shift in gene expression to anaerobic respiration, as determined by microarray analysis, and this shift may have led to the abrogation of photosensitivity by reducing oxidative stress (Nakahigashi et al., 2002). PrfA is a key regulator of pathogenesis in Listeria monocytogenes and is post-translationally regulated such that the protein becomes activated upon bacterial entry into the cell cytosol; prfA mutants that are constitutively activated show impaired motility (Xayarath et al., 2011). Lastly, many scl-L clones were found to be homologous to specific genes of P. multocida that may be involved in virulence, for example, infB, secD, glpT, and tadG. The infA and infB genes encode translation initiation factors 1 and 2 and are essential for the initiation of protein synthesis in prokaryotes (Laalami et al., 1991). The infB gene was picked out in this study, and infA was expressed within macrophages by S. typhi, as identified by SCOTS (Faucher et al., 2005).

In a symbiotic host system, collagen degradation could benefit the bacteria, but would be harmful for the eukaryotic host. Using a polyphasic approach, we investigated the presence of

collagenolytic activity in the bacterial community hosted by the marine sponge Cymbastela concentrica. Functional screening for collagenase activity using metagenomic library clones (227 Mbp) and cultured isolates of sponge’s bacterial community, as well as bioinformatic analysis of metagenomic shotgun-sequencing data (106 679 predicted genes) were used. The results show that the abundant members of the bacterial community contain very few genes encoding for collagenolytic enzymes, while some low-abundance Dabrafenib clinical trial sponge isolates possess collagenolytic activities. These findings indicate that collagen is not a preferred nutrient source for the majority of the members of the bacterial community associated with healthy C. concentrica, and that some low-abundance bacteria have collagenase activities that have the potential to harm the sponge through tissue degradation. Collagen is the major component of extracellular matrices of all metazoan life and represents an important protein conferring integrity and the physical form of eukaryotic organisms

(Harrington, 1996; Exposito et al., 2008). Sponges are among the oldest Metazoa and often contain collagen, which is either dispersed as GSK2126458 in vivo thin fibrils or organized as bundles, termed spongin, in the intercellular matrix (Simpson, 1984; Brusca & Brusca, 1990). The expression of collagen is known to be essential for the development and structural integrity of sponges (Garrone et al., 1975; Shimizu & Yochizato, 1993; Krasko et al., 2000). Sponges harbour specific bacterial communities in different

cellular compartments, often for an extended period of time, and hence close associations between the microorganisms and the sponge host have been established (Taylor et al., 2007). Collagen is an essential and abundant part of the internal mesohyl structure of most sponges AZD9291 (and in particular the Demospongia), where many microorganisms reside. As a rich source of nitrogen and carbon, collagen could provide nutrients for the sponge-associated microorganisms, and this may potentially have implications for the structural integrity of the host. A few cases of sponge diseases have been attributed to the presence of bacterial pathogens (Gaino & Pronzato, 1989; Webster et al., 2002; Mukherjee et al., 2009) and collagenolytic enzymes have been speculated to lead to tissue necrosis in sponges. Generally, bacterial collagenases, including the well-characterized enzymes from Clostridium sp. (Matsushita et al., 1994) and Vibrio sp. (Yu & Lee, 1999; Vaitkevicius et al., 2008), have been linked to pathogenicity and are regarded as virulence factors in human disease.

4). On some occasions the monkeys would have numerous ‘eye-closed’ periods of short duration or only a few eye-closed epochs of extended CYC202 in vitro duration. It was notable that the vast majority of the Type 1 neurons described here had regular firing patterns during sleep, as illustrated for a typical single neuron in Fig. 8. The same property was described by Rolls et al. (2003) for single neurons in the subgenual cingulate cortex BA25 during periods of eye-closure. However, of note is that a few

Type 1 cells showed minor variations in the fine temporal patterning of neuronal firing during some ‘eyes-closed’ epochs, with some exhibiting ‘burst-like’ responses. The quantitative areal distribution of cell Types 1, 2 and 3 neurons in mPFC are given in Table 2 (see also Fig. 1C–E). Finally, it was not possible to ascertain unequivocally whether the neurons being studied electrophysiologically were excitatory projection pyramidal cells or local circuit inhibitory neurones. However, the likelihood is that most of the recorded cells were pyramidal projection neurons

as the spike durations were typically greater than 1.2 ms, which is highly characteristic www.selleckchem.com/products/ganetespib-sta-9090.html of cortical pyramids (Rolls et al., 2003). The principal results of this study indicate that there are two populations of neurons throughout the monkey mPFC that significantly altered their firing rates when the subjects ‘closed’ or ‘opened’ their eyes. Type 1 cells (8.4% of all cells recorded) significantly increased their firing rate when the monkey became drowsy or closed its eyes, whilst Type 2 cells (1.8%) significantly decreased

their firing rate on eye-closure. Together these electrophysiological cell types represent a modest population (10.2%) of all the mPFC neurons screened in this study. Histological reconstructions confirmed that the cells studied electrophysiologically were in BAs 9, 10, 13 m,14c, 24b (dorsal anterior cingulate cortex) and 32 (pregenual cingulate cortex in primates), (-)-p-Bromotetramisole Oxalate with many of the recorded cells being located in the deep layers of the cortex (see Fig. 1C–E). A previous paper from our laboratory reported that neurons in BA25 (subgenual cingulate cortex) of the macaque mPFC also significantly increased their firing rates when monkeys went to sleep (Rolls et al., 2003). Of note is that comparable to the neurons reported here, the cells studied by Rolls et al. (2003) did not respond to gustatory, olfactory and most visual stimuli. Rolls and colleagues also presented evidence of four neurons in the orbitofrontal cortex (BA13) responding in a similar manner. The present study thus confirms and extends to further areas of mPFC the observations of the earlier companion paper. Taken together these two studies indicate that there are distributed populations of neurons throughout the mPFC of monkeys that selectively respond to being either ‘asleep’ or ‘awake’.

Carbon sources were provided as 6 mM [succinate, TSA, TCA, p-sulfobenzoate (PSB) and terephthalate (TER)]

or 3 mM [protocatechuate (PCA)]. Vitamin supplements for minimal media were prepared as described elsewhere (Pfennig, 1978). Soy broth was purchased from Sigma-Aldrich. The fatty acid composition of cell membranes was determined under contract by the German Collection of Microorganisms (DSMZ). 16S-rRNA gene sequences of about 1500 bp were generated following standard procedures. PCR and sequencing were performed using bacterial GSI-IX 16S primers, 27f and 1492r (27f: AGR GTT TGA TCM TGG CTC AG; 1492r: CGG KTA CCT TGT TAC GAC TT) (Weisburg et al., 1991). PCR amplification was performed using the Taq PCR Master Mix Kit (Qiagen) in a total volume of 50 μL, containing 0.5 U μL−1 of Taq DNA polymerase, 1 × Qiagen PCR buffer, 1.5 mM MgCl2 selleck compound and 200 μM

of each dNTP, 10 μM of each primer and 1–10 ng of template DNA. PCR was carried out with an initial denaturation at 95 °C for 3 min, followed by 30 cycles of amplification (denaturation at 95 °C for 1 min, annealing at 55 °C for 2 min and extension at 68 °C for 3 min), with no final extension. Amplified DNA was purified using the QIAquick PCR Purification Kit (Qiagen) and sent to the NERC Biomolecular Analysis Facility (Edinburgh) for sequencing. The alignment of each gene sequence was refined by eye using se-al v2.0a11 (Sequencing Alignment Editor Version 2.0 alpha 11; software available at http://tree.bio.ed.ac.uk/software/seal/). The primer pairs and conditions for PCR mapping of tsa genes were described elsewhere (Tralau et al., 2001, 2003a, b; Mampel et al., 2004). Sequence data were analyzed using standard software (chromas™ from Technelysium

and dna-star™ package from Lasergene). Under the light microscope, cultures of ‘strain TA12’ appeared to be homogeneous, motile rods (2 μm long and 1 μm wide). Selective plates (TSA-salts medium) supported the growth of small (<0.5 mm), homogeneous colonies whose surface was opalescent ochre. However, attempts to sequence the 16S rRNA gene led to reads of poor quality, and the analyses of fatty acids indicated no Cytidine deaminase logical identification; hence, a mixed culture was suspected. Colonies picked from one passage on complex medium required a month to grow to the stationary phase in selective medium, and colonies picked after several passages on complex agar no longer grew in selective medium. Moreover, long incubation on plates of complex medium yielded colonies with different types of edges. Nonetheless, sequencing still indicated mixed cultures. With the assistance of the DSMZ, these mixtures were separated by alternate cultivation on soy agar and in selective medium supplemented with a vitamin solution. As colonies on soy agar looked alike, colonies were picked at random and transferred to a full soy broth and selective medium with vitamins, respectively.

If the live vaccines are administered non-simultaneously and within 4 weeks, it is recommended that the second vaccine administered should be repeated. We report the successful vaccination and generation of a protective immune response to yellow fever (YF) vaccine that was administered to an adult traveler 21 days after receiving another live viral vaccine. A 60-year-old female was seen at the Adult Immunization and Travel Clinic of the San Francisco Department of Public Health 6 days prior to departing on a 2-week visit to western Uganda. She was born and resided in the United States, was in good health, and had no selleck screening library history of

prior flavivirus infection, receipt of YF or Japanese encephalitis vaccinations, or travel to a YF endemic area. The CDC recommends that all travelers ≥9 months of age visiting Uganda be vaccinated against YF.2 Furthermore, at the time of consultation there was even greater concern about the risk of natural infection because of an outbreak of YF occurring in the northern part of the country.3 The client reported receiving an injection of zoster vaccine (Zostavax, Merck Sharp & Dohme, Whitehouse Station, NJ, USA), a live-attenuated viral vaccine, at a pharmacy 21 days earlier. We informed

her that the live zoster vaccine could affect her response to YF vaccine, and that she could be at increased risk of an adverse reaction to YF vaccine due to her age.4 Despite these considerations, and in light of the ongoing outbreak, she agreed with our recommendation in favor of vaccination against YF. We administered YF vaccine (YF-Vax; Sanofi click here Pasteur, Swiftwater,

PA, USA) as well as inactivated vaccines against typhoid, meningococcal infection, and polio (Typhim Vi, Menactra, and IPOL; Sanofi Pasteur). We also prescribed a regimen of daily malaria chemoprophylaxis with atovaquone–proguanil, and instructed her to use prevention measures to reduce her mosquito exposure. She returned to our clinic 5 weeks later, in preparation for a 6-month trip to the same region in Uganda. According to published CDC recommendations, she should have been given a second dose of YF vaccine. However, because her age Nintedanib (BIBF 1120) was a precaution to initial vaccination, and since there was sufficient time to do so, we opted to check her immunity to YF before administering a second dose of the vaccine. A serum specimen was obtained and analyzed at the CDC Division of Vector-Borne Diseases in Fort Collins, Colorado, for neutralizing antibodies against YF virus. At CDC, a 90% endpoint plaque reduction neutralization test (PRNT90) titer of ≥20 is considered protective against YF virus infection.4 Our client had a titer of 1,280 in her serum obtained 35 days after vaccination. Infection with YF virus, a mosquito-borne flavivirus, most commonly is asymptomatic or causes mild febrile illness.

[19] Therefore, information on stool consistency alone was also calculated (a higher number indicated a looser stool). (3) Upper respiratory symptoms: recorded using a modified Jackson system, which detailed the severity of seven items (malaise, chilliness, sneezing, sore throat, runny nose, blocked nose, and cough) each on a 0 to 3 Likert scale (the headache score was removed).[20] As there are no sufficiently

sensitive and specific clinical definitions of presence or absence of upper respiratory infections,[20, 21] the Niemen method of defining presence of upper respiratory symptoms as any score learn more above 1 was used.[22] (4) Anxiety: recorded using the short form state-trait anxiety scale, which recorded the severity of six items on a 0 to 3 Likert scale (adapted from the usual 1–4 scale to ensure consistency with the other self-report measures).[23] Alpha coefficients for the anxiety scale in the present study ranged from 0.83 to 0.92 (above the recommended value for psychological measures of 0.70). (5) Fluid intake: determined daily by using drink bottles of known volume and bead counters to record refills. Total fluid intake was also determined from 24-hour food and fluid

diaries on day 3 (1,100 m) and day 13 (4,700 m), with food and fluid composition determined by computer software (Dietmaster; Z-VAD-FMK datasheet Lifestyles Technologies Inc, Phoenix, AZ, USA). Arterial oxygen saturation and resting heart rate: by finger tip pulse oximeter (9500, Onyx; Nonin, Plymouth, MN, USA), recorded when participants were sheltered from the wind, after wearing gloves and blinded to their results. The lowest and highest values observed over a 1-minute period were recorded and the mean calculated. To achieve the study’s first aim, for each illness, the individual symptom score and the total symptom score were calculated to provide daily expedition mean scores. Statistical Clomifene differences between days were determined by repeated measures analysis of variance. Significant differences

were followed up by Holm–Bonferroni procedures[24] using 1,435 m as the baseline for comparison (the last day of the baseline period that exhibited normal arterial oxygen saturations). Also, for each illness, the expedition’s daily sum of symptom scores (a marker of expedition symptom burden), daily and total expedition incidence (the number of individuals achieving criteria, when available, for clinical diagnosis), and event rates (expressed per 100 person days) were calculated. Participants with missing data were removed from these analyses. To achieve the study’s second aim, longitudinal linear regression analyses were performed using generalized estimation equations.[25] The predictor variables were day of expedition, height gain, upper respiratory symptoms, stool consistency, anxiety symptoms, arterial oxygen saturation, heart rate, and fluid intake.

Three out of four consumers (n=134, 76%) announced that they would value educational material with an integral magnifying Z-VAD-FMK order glass to help them read and understand food labels. There were no significant differences in the findings attributable to the location of interview. It was concluded that the majority of consumers try to lead a healthy lifestyle and eat a healthy diet but find food labels confusing and too small to read. Educational material with an integral magnifying glass may assist consumers in making healthier food choices. Copyright © 2011 John Wiley &

Sons. “
“The global incidence of pregestational diabetes mellitus (PGDM) is on the increase. Pregnancy outcome in these women is much worse compared to those without diabetes, from higher rates of miscarriage, congenital malformations and perinatal mortality. This small audit is a retrospective case note analysis of women with PGDM birthing over ZD1839 price 12 months at a health facility in Australia serving a high-risk and migrant multicultural population. The local prevalence of PGDM was high (0.63%). A large number (56.5%) of the 23 women whose case notes were analysed were older (>30 years) and, of these, 77% were non-Caucasians. Six women were pregnant for the first time. Many (69%) were on preconception folate supplementation. Data on satisfactory pre-pregnancy

glycaemic control (HbA1c > 6.1% [43mmol/mol]) were found in two women and, very though HbA1c was >7.1% (54mmol/mol)- in some, HbA1c readings in all three trimesters were not identified for each woman. Nine women used metformin and insulin was prescribed in the vast majority (82.6%).

Overall, vaginal birth rate was 43% which was even higher (58.8%) among those who attempted vaginal birth, seemingly higher than national figures. Mean gestation at delivery was 37 weeks with four macrosomic (>4.5kg) babies. There was one stillbirth and the neonatal morbidities were in keeping with average. Breastfeeding rates were compatible with the baby-friendly status of the hospital. Following this audit, the provision of antenatal care for this high-risk pregnancy group has been changed in order to improve the quality of care. This is due for re-audit in due course. Copyright © 2012 John Wiley & Sons. “
“In 2010, Leicester City Primary Care Trust commissioned an Intermediate Care Diabetes Service. One aspect of the service plan was to work with the local ambulance trust to gather data around patients using ambulance services for hypoglycaemia, and to provide an advisory service for individuals post ambulance call-out. This audit identified 388 diabetic emergency ambulance call-outs locally (for the period 1 September 2010 to 31 March 2011) including those for hypoglycaemia in the Leicester City area. The new service commissioned by Leicester City included diabetes specialist nurse assessment within two working days for all hypoglycaemic individuals accessing ambulance services.

We have also demonstrated that PamI is responsible for the stable maintenance of pAMI7 and it is also able to stabilize a heterologous replicon. The stabilization effect is most probably caused by a decrease in the level of MTase in the plasmid-less cells after numerous MG-132 manufacturer rounds of cell division. In such a scenario, the remaining MTase becomes insufficient to protect all of the recognition sites on the newly replicated chromosome, which results in cleavage of its DNA by the remaining REase and ultimately the death of the cell (Handa & Kobayashi, 1999). R-M systems can therefore act at the postsegregational

level, which is also a typical feature of plasmid-encoded TA stabilization systems. However, the TA systems, in contrast to the R-M modules, function through the differential stability of the toxin and antitoxin (Dziewit et al., 2007). Bioinformatic analyses revealed that plasmid pAMI7 contains a TA system, whose components show significant similarity to members of the relBE/parDE superfamily. Intriguingly, the results of our previous study strongly suggested that this system is not functional (Dziewit et al., 2011). Therefore, selleck products it is highly probable that the loss of the activity of the

TA system is compensated by the presence of the functional PamI ‘stabilizing’ system. This form of ‘symbiosis’ between an R-M system and its host plasmid may promote the spread, and therefore, the long-term persistence of R-M complexes in a wide range of bacteria (Takahashi et al., 2011). We acknowledge J. Baj for critical reading of the manuscript. This work was supported

by the Ministry of Science and Higher Education, Poland (grants PBZ-MNiSW-04/I/2007 and IP2010 008670). “
“Phosphate metabolism regulates most of the life processes of microorganisms. In the present work Rebamipide we obtained and studied a Streptomyces lividans ppk/pstS double mutant, which lacks polyphosphate kinase (PPK) and the high-affinity phosphate-binding protein (PstS), impairing at the same time the intracellular storage of polyphosphate and the intake of new inorganic phosphate from a phosphate-limited medium, respectively. In some of the aspects analyzed, the ppk/pstS double mutant was more similar to the wt strain than was the single pstS mutant. The double mutant was thus able to grow in phosphate-limited media, whereas the pstS mutant required the addition of 1 mM phosphate under the assay conditions used. The double mutant was able to incorporate more than one fourth of the inorganic phosphate incorporated by the wt strain, whereas phosphate incorporation was almost completely impaired in the pstS mutant.

1A). Thus, presynaptic terminals of granule Sirolimus cells probably express NRX isoforms that could bind to both NL1(−) and LRRTM2. Interestingly, when HA-Cbln1 was applied to HEK293 cells expressing NL1(−),

synaptogenesis was significantly inhibited (Fig. 1A). In contrast, HA-Cbln1 did not affect synaptogenesis observed in HEK293 cells expressing LRRTM2 (Fig. 1A). HA-Cbln1 did not directly bind to HEK293 cells expressing NL1(−) or LRRTM2 (data not shown). LRRTM2 is reported to bind to NRXβ(S4−), which lacks a splice site 4 insert, whereas NL1(−) binds to both NRXβ(S4−) and NRXβ(S4+) (Boucard et al., 2005; Ko et al., 2009). Indeed, presynaptic terminals of cbln1-null granule cells accumulated on HEK293 cells expressing LRRTM2 were preferentially inhibited by NRX1β(S4−)-Fc. In contrast, synaptogenesis induced by NL1(−)cells was preferentially inhibited by NRX1β(S4+)-Fc (Supporting Information Fig. S1). Therefore, we hypothesized that Cbln1 may interact with NRXβ(S4+) expressed at presynaptic sites in granule cells and, thus, specifically interfere with NL1(−)-induced synaptogenesis. To examine this hypothesis, we next expressed GFP-tagged NL1(−) in HEK293 cells and examined whether HA-Cbln1 affected the binding between

NL1(−) and NRX1β(S4+). The extracellular domains of NRX1β isoforms were attached to the Fc fragment of IgG. We confirmed that both NRX1β(S4+)-Fc and NRX1β(S4−)-Fc bound to HEK293 cells expressing find more NL1(−) (Fig. 1B). Application of HA-Cbln1 to the culture medium Ergoloid specifically and significantly reduced the interaction between NL1(−) and NRX1β(S4+)-Fc (Fig. 1B). These results indicate that Cbln1 interacts with NRX(S4+) and competes with the NL1(−)-NRX(S4+) pathway. To confirm that Cbln1 bound to NRX(S4+), we performed cell-based binding assays, which were previously used for the characterization of interaction between GluD2 and Cbln1 (Matsuda et al., 2010). GluD2 served as a positive control, and GluD2 lacking the NTD (GluD2ΔNTD), to which Cbln1 did not bind,

served as a negative control for the binding assays. At 2 days after transfection, cells were incubated with recombinant HA-Cbln1 for 4 h. Immunoblot analyses (Fig. 2A) showed that HA-Cbln1 bound to HEK293 cells expressing NRX1β(S4+) or GluD2, but not to cells expressing GluD2ΔNTD. Immunocytochemical analyses also showed that HA-Cbln1 bound to HEK293 cells expressing NRX1β(S4+), whereas HA-CS-Cbln1, a trimeric complex that did not possess synaptogenic activities (Matsuda et al., 2010), did not bind (Fig. 2B). Although LRRTMs interact with both NRXα(S4−) and NRXβ(S4−) (Ko et al., 2009; de Wit et al., 2009; Siddiqui et al., 2010), certain NL isoforms bind preferentially to β-isoforms of NRXs. Thus, we examined which isoforms of NRXs interacted with Cbln1 in the cell-based binding assays.

The objectives of our study were to evaluate the immunogenicity and the safety of a novel adjuvanted pandemic vaccine in this particular patient population. Answers in this regard are critical, in particular to help elaborate whether squalene-based adjuvants can be administered safely and should be integrated in future seasonal influenza vaccines [7]. Between 16 November and 23 December 2009, a total of 760 immunocompromised adult patients (including 121 individuals with HIV infection) and 138 healthy controls were enrolled in this prospective, open-label, single-centre, parallel-cohort study. Inclusion criteria for HIV-infected patients were a minimum age of 18 years, residence in the area surrounding

Geneva, Small molecule library regular follow-ups being carried out at the University Hospitals of Geneva and a CD4 T-cell count of either >500 cells/μL (group 1) or <350 cells/μL (group 2) to ensure the inclusion of patients with both a better preserved and a more limited T-cell reservoir. Healthy controls were recruited among family members, as immunization was also recommended for relatives of immunocompromised patients. The trial was approved by the institutional review board (ID: CER-09-234), registered on ClinicalTrials.gov prior to patient enrolment (ID: NCT01022905) and conducted in accordance with the principles of the Declaration

of Helsinki, OSI-906 in vitro the standards of Good Clinical Practice, and Swiss regulatory requirements. Written oxyclozanide informed consent was obtained from each subject prior to inclusion. A study

extension was granted for the renewed inclusion of HIV-infected patients during the following 2010/2011 influenza season. According to official Swiss recommendations, healthy subjects received one dose and HIV-infected patients two doses of AS03-adjuvanted split-virus influenza A/09/H1N1 vaccine (Pandemrix®; GlaxoSmithKline) at an interval of 3–4 weeks. Each dose of Pandemrix® contained H1N1 antigen (3.75 μg), squalene (10.69 mg), DL-α-tocopherol (11.86 mg) and polysorbate 80 (4.86 mg). A single vaccine lot was used and administered into the deltoid muscle with a 25-mm needle. During the following 2010/2011 season, influenza immunization consisted of one dose of nonadjuvanted trivalent split-virus influenza vaccine (Mutagrip®; Sanofi Pasteur, Lyon, France) containing the antigens of A/California/07/2009 (H1N1), A/Perth/16/2009 (H3N2) and B/Brisbane/60/2008 at a dose of 15 μg each. Medical information was obtained through a detailed medical history taken at the time of enrolment and completed using the patient’s medical record or via correspondence with the primary care physician. A paper-based case report form (CRF) was designed for automatic data processing and transfer to a virtual data base. Blood was collected on the day of the first immunization and 3 to 4 weeks after the last vaccine dose. Sera were prepared and stored at −20°C until they were used.