The activity of this concentrate was analyzed

by the antimicrobial-activity plate assay using serial dilutions. A growth inhibition zone appeared when microcin N was present. The producer strain E. coli MC4100 pGOB18 was grown in M63 medium for 8 h and the culture supernatant was loaded on the previously activated Sep-Pak C18 column (Waters, Milford, MA). The microcin was eluted with increasing concentrations of methanol/water solutions (0–95%), collecting 1-mL fractions. The purification of SB431542 datasheet microcin N was performed by HPLC. One milliliter of semi-purified microcin obtained from the Sep-Pak C18 column was dried in a SpeedVac. Microcin N was resuspended in 50 μL of acetonitrile solution (40%, v/v) and loaded in an HPLC Beckman Gold System (Beckman Coulter Inc., Brea, CA). The sample was chromatographed in an isocratic condition using 40% v/v acetonitrile as the mobile phase at a flow rate of 1 mL min−1 on a Beckman ODS column (5 μm × 4.6 mm × 25 cm) (Beckman Coulter Inc.). Proteins were detected at 215 nm using a Beckman System Gold 166 Detector. The fraction corresponding to microcin N was PD-1/PD-L1 targets identified using sensitive plate assay. The mass of microcin N purified by HPLC was determined on a Microflex MALDI-TOF (Bruker Daltonics Inc., MA). The spectra were performed under the positive ion mode, averaging 10 spectra obtained by 40 laser shots each. Fluorescence labeling of microcin N was achieved according to the method

described by Ragland et al. (1974). Briefly, 300 μL of purified microcin N was concentrated to 10 μL by evaporation. This concentrate was then mixed with 4 μL of

0.4 M borate (pH 9.0) and 8 μL of fluorescamine solution (2 mg mL−1 in dimethyl sulfoxide). After 1 min of reaction at room temperature, 7 μL of loading buffer was added. Samples oxyclozanide were denatured by boiling for 5 min and then analyzed by Tricine–sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) (Schägger & von Jagow, 1987) to separate polypeptides from 1 to 100 kDa. The DNA sequences reported in this paper are deposited in GenBank under accession number FJ895580. To confirm the accuracy of the previously reported sequence of the genetic system of microcin 24 (now microcin N), we sequenced the entire fragment cloned into pGOB18. The previously reported sequence (GenBank accession number U47048) has one deletion and three insertions with respect to the sequence reported in this work. These differences resulted in important changes in the putative regulator gene and in the structural gene for microcin N. Because of the differences with the previously reported sequences, we decided to rename these genes in order to use the commonly accepted microcin nomenclature as shown in Fig. 1. The mcnR gene encodes for a putative regulator of 144 amino acids and shares 99% (143/144) identity with proteins ACA51174 from S. enterica ssp. enterica serovar Dublin and ABZ89587 from the E. coli conjugative plasmid pOLA52 (Norman et al., 2008).

It has been strongly suggested that physical and/or MP of a movement sequence improves performance and induces plasticity in the cerebellum (Jenkins et al., 1994; Toni et al., 1998; Lacourse et al., 2004). Strangely, anodal tDCS over the right cerebellar hemisphere impaired the motor performance. Similarly, a former study using anodal tDCS over the cerebellum showed that anodal tDCS impaired the practice-dependent proficiency increase in working memory (Ferrucci et al., 2008). Galea et al. (2009) found that anodal tDCS over the right cerebellar cortex Selleck Osimertinib can increase the inhibitory tone that the cerebellum exerts over the M1. The inhibition of the M1 after

cerebellar tDCS could be one explanation for the impairment of handwriting legibility observed in our study. Potential Selleckchem Palbociclib limiting aspects of the study should be mentioned. (i) In principle, motor practice alone of the handwriting task with the non-dominant hand over six experimental sessions could have had an impact on motor performance and it might have somewhat compromised the interpretation of the results. However, this is improbable in our opinion as the experimental session order was counterbalanced among subjects and baseline writing performance on the experimental first day did not differ from that on the last day, (ii) It cannot

be ruled out that additional cortical areas may have been influenced by tDCS due to the relatively poor spatial resolution of the technique (Nitsche et al., 2008; Datta et al., 2009). Although we cannot Reverse transcriptase completely rule out this possibility, it should be noted that other studies using tDCS successfully modulated close cortical areas in different ways (Nitsche & Paulus, 2000; Nitsche et al., 2003b; Vollmann et al., 2012). (iii) Some studies have reported gender differences in responses to tDCS (Knops et al., 2006; Boggio et al., 2008; Chaieb et al., 2008). In the present study, as the most of subjects were women, it is possible that sex hormones somewhat influenced the results of our study. It is necessary to replicate the study using male participants in future research to investigate

a potential gender influence on the results. In conclusion, our results suggest that MP-induced effects in improving motor performance can be successfully consolidated by excitatory non-invasive brain stimulation on the M1 and left DLPFC. Although this finding is novel, further investigation is needed to understand how motor performance improvement is consolidated after mental training and whether it can be extended to other populations such as patients with neurological pathologies. If so, tDCS could be effectively used as a complementary method to increase the mental training effects. Moreover, our findings may help to improve to understanding about the specific role of each area involved in the MP effects on motor learning. However, a better understanding of the action mechanisms is essential for MP to be used effectively as a therapeutic tool.

Our results indicate

that L. fermentum NTD are distributed not only in the cytoplasm but also on selleck compound the cell wall surface, and further studies showed that surface-attached NTD can be released into the culture broth and conventional buffers. Lactobacilli can be divided into two groups depending on whether or not they require deoxyribonucleosides for growth (Kaminski, 2002). Most lactobacilli that utilize the salvage pathway degrade exogenous nucleosides to the nucleobase and pentose sugar via a nucleoside phosphorylase. Others possess a special salvage system based on a nucleoside deoxyribosyltransferase and require a deoxynucleoside in combination with purine and pyrimidine bases for their DNA synthesis (Kilstrup

et al., 2005). N-deoxyribosyltransferases (EC 2.4.2.6), also called trans-N-deoxyribosylases, catalyze the transfer of a 2′-deoxyribosyl group from Metformin a donor deoxynucleoside to an acceptor nucleobase (Anand et al., 2004). This enzyme was initially described for lactobacilli and has also been found in certain species of Streptococcus (Chawdhri et al., 1991) and in some protozoans such as Crithidia luciliae (Steenkamp, 1991). Two types of N-deoxyribosyltransferase have been described in lactobacilli: type I is purine deoxyribosyltransferase (PTD), specific for the transfer of deoxyribose between two purines; type II is nucleoside 2′-deoxyribosyltransferase (NTD), which catalyzes the transfer of deoxyribose between either purines

or pyrimidines (Holguin & Cardinaud, 1975; Miyamoto et al., 2007). Several dozen reports on lactobacilli N-deoxyribosyltransferase have been Thalidomide published since the initial study by Macnutt (Macnutt, 1950). The three-dimensional structure of these enzymes has been solved, and their kinetic mechanisms as well as their catalytic and substrate binding sites have been well characterized (Armstrong et al., 1996; Anand et al., 2004). The transfer reactions, catalyzed by either PTD or NTD, proceed following a ping-pong bi-bi mechanism by formation of a covalent deoxyribosyl enzyme intermediate (Danzin & Cardinau, 1974; Danzin & Cardinaud, 1976). As NTD has broader substrate specificity than PTD, it has attracted more attention. NTD also has a hydrolase function such that, in the absence of an acceptor base, the nucleoside is converted to its base and deoxyribose (Smar et al., 1991). Most antiviral or anticancer drugs are analogues of naturally occurring nucleosides. The use of purified enzyme or intact bacterial cells containing NTD enables a one-pot transglycosylation reaction at high yields, providing an interesting alternative to traditional multistep chemical methods (Fernandez-Lucas et al., 2010). Stereospecific reactions and high tolerance for various modifications in the bases also make NTD ideally suited to serve as biocatalyst for the production of nucleosides and nucleoside analogues (Okuyama et al.

We also isolated and characterized the filament–hook–basal body of the polar flagellum. The proteins in this structure were analyzed by MS. Eight internal

sequences matched with known flagellar proteins. The comparison of these sequences with the protein database from the complete genome of V. shilonii allows us to conclude that some components of the polar flagellum are encoded in two different clusters of flagellar genes, suggesting that this bacterium has a complex flagellar system, more complex possibly than other Vibrio species reported so far. Motility provides a survival advantage under a wide variety of environments, allowing bacteria to compete successfully for nutrients. Hence, microorganisms have developed a multiplicity of motility systems that allow them to move about in liquid or viscous media and over OSI-906 chemical structure surfaces. Bacteria and Archaea use flagella for locomotion. These are highly complex and efficient structures that not only propel the cell but also play an important role in biofilm formation, adhesion to surfaces and contribute to the virulence process in pathogenic species (for a review, see Kirov, 2003). The bacterial flagellum is formed by a helical filament, which is attached to the cell through a flexible joint known as the hook. The hook 17-AAG datasheet is connected to a complex structure known as the basal body that

spans the inner membrane, the cell wall and the outer membrane (for a review, see Berg, 2003). A limited number of bacteria possess dual flagellar systems, a polar flagellum for swimming in liquid medium and lateral flagella for swarming that involves translocation on solid surfaces. In various species of Gram-negative marine Vibrio such as Vibrio parahaemolyticus, Vibrio alginolyticus and Vibrio harveyi, the single-sheathed polar flagellum is constitutive whereas lateral flagella are inducible. However, this is not a general trait for the genus because Vibrio vulnificus, Vibrio anguillarum, Vibrio fisheri and Vibrio 3-oxoacyl-(acyl-carrier-protein) reductase cholerae do not possess a lateral flagellar system (for a review, see McCarter, 2001, 2004; Merino et al.,

2006). Induction of lateral flagella occurs in response to growth on surfaces or highly viscous media; this process is mediated apparently by the sodium-driven polar flagellar motor, which acts as a mechanosensor (Belas et al., 1986; McCarter et al., 1988; Kawagishi et al., 1996; Merino et al., 2006). Upon an increase in viscosity or contact with a surface, rotation of the polar flagellum is hindered and cells differentiate into swarmer cells. In some species, swarmer cells are elongated, multinucleated and hyperflagellated, such as V. parahaemolyticus, Proteus mirabilis and Serratia liquefaciens (Harshey, 1994; Eberl et al., 1999). In contrast, Aeromonas spp. and Azospirillum spp. do not show cell elongation (Merino et al., 2006). Rotation of the flagellar motor is powered by transmembrane ion gradients in V.

In this era of financial austerity, we do

not believe that the 75-fold cost differential (based on a 14-day course for a 70-kg adult at NHS list price including VAT) between AmBd at 1 mg/kg/day (£4.66/50 mg vial, 2 vials/day × 14 = £130.37) Vorinostat and AmBisome at 4 mg/kg/day (£116.03/50 mg vial, 6 vials/day × 14 = £9746.52) is justifiable for HIV-infected patients with normal baseline renal function and no other nephrotoxic drugs. Even use of AmBd in the first week, before switching to AmBisome, would incur a cost saving of £4808 per patient treated. Pharmacy departments can stock both preparations and support their safe prescribing by brand name. As an oral alternative to AmBd, UK guidelines are again at odds with IDSA and WHO in recommending fluconazole at the low dose of 400 mg/day, combined with 5FC. Fluconazole is a fungistatic drug associated with worse outcomes when used in initial treatment of CM [9]. Phase II trials have shown improved cryptococcal clearance

and good tolerance using doses of fluconazole up to 1200 mg/day, without or including 5FC [10-12], a combination endorsed by WHO for areas where AmBd cannot be safely administered [3]. Lastly, in the management of raised intracranial pressure (ICP), we agree with recommendations regarding CSF manometry and repeat lumbar punctures, but, given the usual resolution, with appropriate management, of high ICP within the first weeks of induction therapy, would favour use of temporary lumbar drains over shunts in situations of high ICP unresponsive to daily lumbar punctures Romidepsin [13]. In light of these arguments, we would urge the panel to reconsider their recommendations for these aspects of management of patients with CM in the UK. “
“The risk of mother-to-child transmission of HIV can be significantly reduced by giving antiretroviral drugs to both mother and child,

by an appropriate mode of delivery, and by avoidance of breast feeding [1]. However, despite routine antenatal HIV screening and high uptake of interventions to reduce mother-to-child Baricitinib transmission in the UK, potentially preventable mother-to-child transmission of HIV still occurs [2]. To try to avoid potentially preventable infection, a review of local guidelines for managing infants born to HIV-positive women was performed in the North West Perinatal and Paediatric HIV Network. Information on which maternity units in the North West of England and North Wales had delivered HIV-infected women during the years 2006–2009 (296 deliveries; two infants HIV-infected) was obtained from the National Study for HIV in Pregnancy and Childhood (NSHPC) [3]. A questionnaire was sent to each of these units, requesting a copy of their local guidelines. Local guidelines were then compared with the British HIV Association/Children’s HIV Association (BHIVA/CHIVA) guidelines for the management of HIV infection in pregnant women [1].

, 1996). In the genus Flavobacterium, several new species have been described with a rather high 16S rRNA gene sequence similarity, for example Venetoclax the type strains of Flavobacterium weaverense and Flavobacterium segetis share 98.9% 16S rRNA gene sequence similarity, and yet, they have a DDH value of only 34% (Yi & Chun, 2006). Because protein-encoding genes are generally less conserved (Ochman & Wilson, 1987), they may be more appropriate for phylogenetic analysis of closely related species.

Several protein-encoding genes such as glnA, recA and hsp60 have been used for typing and taxonomical purposes within genera in the Bacteroidetes (Gutacker et al., 2002; Sakamoto et al., 2010). In this study, the gyrB gene, encoding for the B subunit of the DNA gyrase, was selected because it was previously used successfully to distinguish between closely related taxa affiliated with the genus Flavobacterium Sunitinib (Suzuki et al., 1999, 2001). Izumi et al. (2003) reported on the use of gyrB primers in a PCR-restriction fragment length polymorphism analysis for the genotyping of F. psychrophilum, and Suzuki et al. (1999) designed gyrB primers to study the phylogenetic relationship for the genus Marinilabilia (Bacteroidetes) and related taxa. We tested all primers reported in these studies in silico on the gyrB sequences available from related genera and from the complete genome of F. johnsoniae DSM 2064

and found considerable mismatches with all groups included in the comparison. Therefore, more general primers were designed based on the available sequence Phospholipase D1 information. As expected

for a more variable housekeeping gene, the distance between the Flavobacterium groups and the type strains is significantly higher in the gyrB gene dendrogram (Figs 2 and S2) in comparison with the 16S rRNA gene dendrogram (Figs 1, S1 and Table 3). The threshold for species definition has been suggested to be 98.7–99.0% 16S rRNA gene sequence similarity byStackebrandt & Ebers (2006), hereas for the gyrB phylogeny, this is less well documented. Suzuki et al. (2001) reported that the proposed limit for species identity, the 70% DNA reassociation value, corresponds to 88.8%gyrB sequence similarity in the subset of the Bacteroidetes they studied, whereas several other studies revealed a wide range of interspecies similarity values [60.0–89.0%gyrB gene sequence similarity within the genus Helicobacter (Epsilonproteobacteria) (Hannula & Hanninen, 2007), 75.4–95.0% within the genus Bacillus (Firmicutes) (Wang et al., 2007), 85.0–97.5% within the genus Aeromonas (Gammaproteobacteria) (Yanez et al., 2003), 77.5–97.6% within the genus Gordonia (Actinobacteria) (Kang et al., 2009), 89.5–98.2% within the genus Kribbella (Actinobacteria) (Kirby et al., 2010) and 70.1–98.7% within the genus Streptococcus (Firmicutes) (Itoh et al., 2006)].

We found that vpsL mRNA levels were approximately 15-fold higher in the strain with increased NspC levels (Table S1). These results indicate that increased NspC levels affect biofilms through a vps-dependent mechanism. Moreover, because the measurements in these 17-AAG assays are normalized to cell density and an internal standard, respectively, they support the conclusion that the effect of NspC on biofilms is in addition to and independent of its effect on growth. In most cases, biofilm formation and motility are inversely regulated

such that signals or mutations that lead to increases in biofilm formation lead to decreases in motility (Watnick et al., 2001; Moorthy & Watnick, 2005). To determine the effect of increased NspC levels on motility, check details we performed motility assays using semisolid agar plates. Increased NspC levels led to a twofold decrease in the swarm diameter (Fig. 1d), indicating that increased NspC levels affect biofilms and motility in an inverse manner. Because

decreases in intracellular norspermidine levels lead to a decrease in biofilm formation in V. cholerae O1 El Tor, we hypothesized that the increase in biofilm formation may be a consequence of increased levels of norspermidine in these cells. To test this hypothesis, we extracted and quantified the cellular polyamines from shaking cultures grown to log phase (Fig. 2a and b). Norspermidine levels did not increase in the strain overexpressing nspC. Under the conditions of our experiment, V. cholerae Galactosylceramidase also contains significant amounts of putrescine, diaminopropane, spermidine, and a small amount of cadaverine. The levels of these polyamines were also not different between the two strains. Next, we quantified polyamines in the planktonic cells and biofilm-associated cells of static biofilm cultures. Again, we observed no differences in the levels of the various polyamines in the two strains (Fig. 2c and d). However, cadaverine levels were increased in both the planktonic and biofilm-associated cells as compared

to shaking cultures. These results indicate that the increased biofilm levels seen in this strain did not result from increased levels of norspermidine or changes to levels of other polyamines in the cells. We next hypothesized that cells could indeed produce increased amounts of norspermidine as a result of increased NspC levels; however, the excess norspermidine might be exported to maintain norspermidine homeostasis in the cell. To test this hypothesis, we quantified the polyamines in the spent media of these strains as well as sterile LB medium. We did not detect any norspermidine in LB or the spent media. In addition, we found that the spent medium of these strains contained putrescine, diaminopropane, cadaverine, and spermidine; however, only putrescine levels were higher in the spent media of either of the strains as compared to LB, in shaking cultures (Fig. 3).

1 °C s−1 to 95 °C. Fluorescence was monitored at regular intervals during the extension step and continuously during the melting. The experiment was

completed in approximately 45 min. The target sequence is detected when the fluorescence curve MK-1775 concentration turns abruptly upward above the threshold. Each DNA sample is characterized by this point of the curve, called the crossing point (Cp). The specificity of the primers tested on type strains was then validated using DNA extracted from a set of 11 Aspergillus section Flavi strains, two other Aspergillus species and six fungal genera commonly found in the environment (Table 1). Within the section Flavi, PCR results were compared with the identification data obtained by means of the calmodulin gene sequencing as described previously (O’Donnell et al., 2000). Three RAPD analyses were performed as described by Yuan et al. (1995) with the primers OPA-04, OPB-10, OPR-01, and sequences AATCGGGCTG, CTGCTGGGAC and TGCGGGTCCT, respectively. DNA amplification was carried out in a final volume of 25 μL containing 100 ng of template DNA, 5 pmol of primer (Sigma-Aldrich), 1 U of Taq DNA polymerase (Sigma-Aldrich), Daporinad cost 1 × of Taq DNA buffer (Sigma-Aldrich), 100 μM of dNTPs and 1.5 mM MgCl2. Amplification was performed

in a thermocycler (Biometra, Tgradient, Göttingen, Germany) and the amplified products were separated by gel electrophoresis according to Yuan et al. (1995), except that the gel was stained with GelRed™ (Biotium Inc., Hayward, CA). One microgram of DNA was digested with SmaI (Klich & Mullaney, 1987) under the following conditions: overnight incubation at 25 °C in a final

volume of 25 μL containing 1 U of SmaI (Roche Diagnostics GmbH) and 1 × of buffer. Restriction was fractionated by electrophoresis on a 0.7% agarose gel stained with GelRed™ (Biotium Inc.). Two primers, Afaflt-F (forward) and Afaflt-R (reverse), were designed Silibinin on a region of the aflT gene presenting a low level of homology between A. flavus, A. oryzae and other four species of the section Flavi for which the gene sequences were available in GenBank (Fig. 1a). A second primer set, Anits-F (forward) and Anits-R (reverse), was designed on a region of the A. nomius ITS1–ITS2 region unique to this species (Fig. 1b). Before PCR amplification, the theoretical specificity stringency of the primers designed for species of the Aspergillus section Flavi was evaluated using the basic local alignment search tool (blast, NCBI). For each set, no fungal species other than the target Aspergillus species were proposed, i.e. A. oryzae and A. flavus for Afaflt-F/Afaflt-R and A. nomius for Anits-F/Anits-R. Different times and annealing temperatures were tested to define the optimal conditions required for each primer set specificity.

Amongst military personal an association has been found between contracting malaria16 and failure to complete post-travel courses, and in a survey of backpackers 30% were found to have stopped selleck kinase inhibitor medication prematurely.17 Travelers and prescribers agreed that effectiveness concerns about side effects, previous experience, and convenience of doses were the most important reasons for the choice of antimalarial. HCPs are recommended to take these factors into consideration when discussing appropriate malaria chemoprophylaxis with travelers to improve overall adherence. Travelers chose their antimalarial chemoprophylaxis

as part of their usual consultations, and this study was not designed to look at any particular interventions to influence choice or to identify why a particular antimalarial

was chosen. A study of 1,073 Swiss travelers demonstrated the value of detailed written information on informing choice and that adverse effect profiles, previous use, and cost were the most important factors.18 There did not seem to be any characteristic of the traveler, such as length of travel and reason for traveling, determining choice of antimalarial other than those receiving Dxy tended to be younger. This may be related to the cost, where younger backpackers may self-select for the somewhat cheaper see more Dxy regimens. These observation are only related to the decisions made by those traveling <28 days and may differ for those traveling longer term. This study supports the assumption that the 1 week antimalaria post-exposure course using At+Pro could be preferable to a 4-week course with Dxy to encourage 5-Fluoracil datasheet adherence to the prescribed regimen. Further work is required to identify the variety of factors that determine adherence to antimalarials. We would like to acknowledge Professor Robert Horne for his help and advice on this project. We would also like

to thank the study staff at MASTA, NOMAD travel clinics, and the Royal Free Hospital. The study was commissioned and paid for by GlaxoSmithKline. L. R. and A. M. are employees of GlaxoSmithKline. L L G. is the Superintendent Pharmacist and the Director of Nomad Travelstore Ltd. “
“Background. Malaria continues to be a serious, world-wide infection. Atovaquone-proguanil is one of the prophylactic agents recommended for travelers to endemic regions. However, little information is available regarding adherence with this medication. A large proportion of malaria cases reported from travelers is due to non-adherence to prescribed regimens. This study was undertaken to analyze adherence with atovaquone-proguanil prophylaxis and specific factors contributing to non-adherence. Methods. Men and non-pregnant women ≥18 years of age were eligible for inclusion. Enrolled travelers received a prescription for atovaquone-proguanil prophylaxis and were contacted by telephone within 3 weeks of return to the United States.

There was also a weak association of low maternal CD4 cell count and cigarette smoking with an increased risk of prematurity, both in the univariable and in the multivariable analyses. However, confidence intervals on the estimates were wide for both cART and CD4 cell count, and also for nicotine use. Maternal age, ethnicity, history of drug use

and viral load were not associated with the risk of prematurity (Table 2). There is controversy as to whether exposure to cART in HIV-1-infected pregnant women selleck inhibitor increases the rate of premature birth. It has specifically been argued that heterogeneity in outcomes of previous studies [1–3,8] may have been caused by uncontrolled confounding from maternal risk factors in studies indicating elevated risk of prematurity, which included the initial study based on data from the Swiss MoCHiV cohort [1]. Here we reanalysed the updated Swiss data, which provided more complete and precise information on potential risk factors for prematurity, the exact duration and composition of ART, and maternal lifestyle characteristics following the integration of the MoCHiV in the SHCS. In agreement with our earlier combined KU-57788 mouse study with the ECS [2], we found a positive association between intensity of ART regimen (increasing in intensity from no ART to mono/dual ART regimen

to cART) and the risk of premature birth in a crude analysis of all available pregnancies (analysis 1). Prematurity rates increased with calendar year in HIV-1-infected pregnant women, as shown in other studies [9]. This trend was in accordance with the increasing intensity of ART regimen

with calendar year, but not with changes in key maternal risk factors for prematurity. Consistent with intensified ART, pregnant women showed an increase in the median CD4 cell count and a decline in the proportion of women with detectable viral load, while the prevalence of tobacco use and IDU declined. Further analyses (analyses 2 and 3) that included women exclusively on cART and with precise information about the time-point of initiation of treatment indicated higher prematurity rates in children whose mothers started cART before pregnancy as compared with Niclosamide mothers starting in the third trimester, similar to the findings of our previous ECS/MoCHiV study [2]. However, the risk of prematurity was not different between mothers starting cART before and during pregnancy, and there was also no association between the total duration of cART before delivery and the duration of pregnancy. As a consequence of incompleteness of data in the early MoCHiV database, our crude time trend analysis of potentially confounding risk factors for prematurity in all pregnancies (Fig. 2) has to be interpreted with caution. For instance, available information on maternal smoking was imperfect, because no indication of tobacco use may also signify that no information about smoking behaviour was available.