Transdermal delivery (Williams, 2003 and Barry, 2001) offers one

Transdermal delivery (Williams, 2003 and Barry, 2001) offers one potential means of overcoming many of the problems associated with systemic delivery of bacteriophages. Clearly bacteriophages, being viruses rather than small relatively lipophilic drug molecules, do not satisfy the criteria for efficient Anti-diabetic Compound Library manufacturer transdermal absoprtion. Nevertheless, the transdermal delivery of these potent therapeutic agents is of particular interest, as it may overcome many of the problems associated with conventional

delivery methods. To date, transdermal delivery of bacteriophages has not been considered. However, novel microneedle technologies, developed by our Group and others, have now made this a possibility, particularly for thermolabile biomolecules and biological entities (Donnelly et al., 2010a, Donnelly et al., 2010b, Donnelly et al., 2011, Mikolajewska et al., 2010, Migalska et al., 2011, Prausnitz, 2004 and Garland et al., 2011). In this paper, we report for the first time, design and evaluation of a novel hollow polymeric microneedle device for transdermal bacteriophage delivery. T4 bacteriophage ATCC® B11303 and host strain Escherichia coli 11303 ATCC® 11303 were purchased from LGC standards, Middlesex, UK. Luria Bertani (LB) agar was purchased from Sigma–Aldrich,

Dorset, UK. Stock phage solutions were stored at 4 °C and protected from light. E. coli almost was frozen with cryoprotectant beads and glycerol and stored at −60 °C. Isoflurane inhalation anaesthetic was obtained from Abbott Laboratories Ltd., Kent, selleck inhibitor UK. All other chemicals used were of analytical reagent grade. Microneedles (MNs) were manufactured using a prototype micromoulding process. Mould cavities and inserts were micro-machined from brass and inset pins were machined from H-13 tool steel using a specialized Electric Discharge Machining (EDM) process. The moulds were run

on an Arburg 221 KS Allrounder moulding machine. MNs were manufactured from PC. The prototype array of MNs consisted of seven needles at 3 mm centers on a 21 mm × 21 mm base. The MNs were 1 mm in height with a 100 μm off-centre through-hole. The aspect ratio was 1.6:1. The tip sharpness of the prototype needles was approximately 25 μm in radius. The MN array was ultrasonically welded to a reservoir array of the same material as the MN array consisting of a 5 μl reservoir well for each MN. A silicone sealing gasket was used in-between the MN array and reservoir array. To observe MN morphology, images of the MNs were taken using a Leica DC150 digital microscope (Leica, Wetzlar, Germany). MNs were attached to aluminium stubs using double-sided adhesive and coated at 2.5 kV, 18 mA with gold for 45 s (POLARON E5150, Gold Sputter Coater, Quorum Technologies, East Sussex, UK).

However, this does not explain why family size was not related to

However, this does not explain why family size was not related to MMR: there was a wider range of family sizes in the MMR group (parents with a maximum of six children in the family) than in the dTaP/IPV group (with a maximum of only

four children in the family). For MMR, it is possible that apprehensive parents may want to make separate decisions for each child based on information available SB431542 at the time; as found in some of the interviews with parents of preschool children [4]. Alternatively, there may be a critical number of children beyond which any additional children make no difference so that the different family sizes in the two groups gave an artificial result. Clearly, however, a larger study would be needed to investigate these assumptions further. Although subjective norm was found to correlate with intention, it was interesting that this did not predict parents’ intentions to immunise with either MMR or dTaP/IPV. This suggests that friends, family and healthcare professionals did not directly influence parents’ immunisation intentions, even click here though in the interviews most parents said that they would attend for immunisation

because it was ‘the norm’ [3] and [4]. Thus, although parents may discuss their vaccine decisions with these ‘significant others’, these discussions may not directly influence their child’s immunisation status. Indeed, Bennett and Smith [9] found that there was no difference in the families’ or friends’ perceptions of the value of immunisation between those caregivers who had fully vaccinated a child against

pertussis, partially completed the course or refused pertussis vaccination. This finding is also supported by earlier TPB-based research which found that subjective norms were unrelated to immunisation status [13] and [14]. Moreover, across studies and across a range of health behaviours, attitude and perceived control generally emerge as stronger predictors of intention Rolziracetam [19]. Hence, the findings of the present study suggest that, when it comes to preschool immunisation, other factors are more salient than the views of ‘significant others’. Overall, the predictors identified in the regression analyses accounted for 48.0–64.4% of the variance in parents’ intentions to immunise with a second MMR and 52.1–69.5% of the variance in parents’ intentions to immunise with dTaP/IPV. This is consistent with the finding that attitude, subjective norm and perceived control generally account for 40–50% of the variance across studies and health behaviours [19]. Prediction rates were impressive, with 84.0% and 83.7% of parents correctly classified for MMR and dTaP/IPV, respectively. Therefore, by including attitudes and beliefs identified in interviews with parents, the IBIM generated highly predictive models of uptake.

S , M S , S W ); Analysis and interpretation of data (E S , U W ,

S., M.S., S.W.); Analysis and interpretation of data (E.S., U.W., C.F., G.Q., M.S., S.W.); Preparation of manuscript this website (E.S., M.S., S.W.); Critical revision of manuscript (E.S., C.F., G.Q., M.S., S.W.); Final approval of manuscript (E.S., F.D., M.M., P.K., U.W., C.F., D.G., G.Q., M.S., S.W.); Data collection (U.W., C.F., S.W.); Provision of materials, patients, or resources (E.S., F.D., M.M., P.K., U.W., C.F., D.G., G.Q.). The authors thank Fiona Powell, Facilitate Ltd, Brighton, UK, for editorial assistance in the production of this manuscript. “
“Figure options Download full-size image Download high-quality image (1025 K) Download

as PowerPoint slideS. Arthur Boruchoff, MD died May 28, 2013 of cardiac complications. He was born in 1925 in Boston, Massachusetts. He went to high school at Boston Latin School and later received the AB degree from Harvard

College (1945), an MD (AOA) from Boston University (1951), and an MSc (in Ophthalmology) from New York University (1956). Arthur served in the US Navy and was stationed in Japan. His residency was at New York Eye and Ear Infirmary (1951-1956), on the service of Conrad Berens, MD, during which time he spent a year with the US Public Health Service, studying the vitreous body, supervised by Sir Stewart Duke-Elder and Professor Norman Ashton at the Institute buy C646 of Ophthalmology, London (1954-1955). Returning to Boston, he began a clinical practice primarily concentrating in cornea and external diseases. In 1958, he became an Assistant Professor in Ophthalmology at Harvard and rose in rank through the years, becoming an Associate Professor in 1990 at Harvard and finally a Professor at Boston University. Paralleling this, Arthur served on the staff

at Massachusetts Eye and Ear Infirmary (MEEI) and became a Surgeon Oxalosuccinic acid in Ophthalmology in 1974. He served on the Committee on Continuing Ophthalmic Education, 1986-1989, and was on the Board of Surgeons at MEEI (1978-1984), and was Director of and President of the Medical Staff (1987-1988). Dr Boruchoff joined the first Corneal Service and Fellowship in the United States that was founded at MEEI in1960 by Claes Dohlman, MD, PhD with Edward Sweebe, MD. Arthur and Dr Dohlman performed all of the corneal transplants on the Corneal Service. Thus began a 35-year-long association with MEEI and with Claes Dohlman. Dr Boruchoff authored some 76 publications, primarily in the area of corneal diseases and surgery, with special interest in the corneal dystrophies, most co-authored with Dr Dohlman and the MEEI Fellows. The topic of his Castroviejo Medal presentation before the American Academy of Ophthalmology (AAO), from which he received both Honor and Senior Honor Awards, was “Unusual Aspects of the Corneal Dystrophies” (1988). He presented several named lectures, including the John McCullough Lecture at the University of Texas, Galveston (1979) and the Albert C Snell Lectureship, Rochester, New York (1994).

, USA) were coated with 100 μL of washed bacteria (both MAP and M

, USA) were coated with 100 μL of washed bacteria (both MAP and MAA; 1 × 108 cfu/mL), diluted in sodium bicarbonate buffer

pH 9.6 for 60 min at room temperature, while shaking at 300 rpm on a electronic DAPT MTS shaker (IKA Werke, Germany). All subsequent incubations were performed for 30 min shaking at room temperature. After each incubation step, plates were washed three times with PBS containing 0.01% Tween 20. The secondary antibody was goat anti-Mouse (GAM)-PO (Roche, the Netherlands) 1:2000. Peptide ELISA was used for the initial epitope mapping of the monoclonal antibodies generated against MAP Hsp70. The peptide ELISA using cys-linked peptides has been described previously [23]. The different cys-linked peptides were diluted in 0.1 M Tris–HCl, pH 8.0 at a concentration of 15 μg/mL, and 100 μL was added at each well. To study Lenvatinib whether monoclonal antibodies bind to intact bacteria, indicative of the presence of MAP Hsp70 in the bacterial cell wall, suspensions of MAA strain D4 and MAP strain 316F (generous gifts from D. Bakker, CVI) were prepared from log phase liquid cultures. Suspensions of MAA and MAP (both 1010 bacteria/mL in PBS) were diluted 1:100, washed three times by centrifugation (1 min at 14,000 RPM in an

Eppendorf centrifuge (Eppendorf, Germany)) and resuspended in PBS. These suspensions were diluted 1:100 in PBS supplemented with 1% BSA and 0.01% sodium azide (both from Sigma Aldrich, USA) and divided in volumes of 100 μL. The Hsp70 specific monoclonal antibodies were added in a concentration of 5 μg/mL. After incubation for 25 min at room temperature (RT) and three washes with PBS supplemented with 1% BSA and 0.01% sodium azide (FACS buffer), FITC-labelled Goat anti-mouse antibodies (Becton-Dickinson, USA) were added and incubated for 25 min at RT. After three more washes, 10,000 bacterial cells were used for analysis by FACScan (Becton-Dickinson,

USA). Multiplex peptide specific antibody measurements were performed using biotinylated peptides linked to avidin coated fluorescent microspheres (LumAv, Luminex, USA) on a Luminex 100 platform according to instructions Edoxaban provided by the manufacturer (Luminex). A total of 2.5 × 105 beads (100 μL) per uniquely labelled beadset were washed twice with PBS, and subsequently incubated with 10 μmol biotinylated peptide for 10 min at 20 °C. After two washes with PBS, the beads were resuspended in their original volume (100 μL) using PBS supplemented with 1% bovine serum albumine (Sigma Aldrich, USA) and 0.01% sodium azide, and stored in the dark at 4 °C until further use. For multiplex analysis 20 μL of resuspended coated beads of each of up to 20 unique beadsets were pooled in an eppendorf container. To the final volume of beads, the same volume of PBS was added, and mixed. In a round bottom 96 well microtiter plate, 10 μL of the mixed beads was added per well. Subsequently, 100 μL of goat or calf serum per well was added.

A p value ≤ 0 05 was deemed to be statistically significant A

A p value ≤ 0.05 was deemed to be statistically significant. A

paired t-test with Bonferroni correction was used (with p = 0.05/6 = 0.0083) for the pair-wise comparison in muscle activity and marker displacement in the frontal and sagittal planes for the two feedback conditions. Nineteen participants were recruited from the Department of Physical Therapy, Yonsei University, ABT 199 Korea. The characteristics of the participants are presented in Table 1. All participants completed all aspects of the testing procedure according to the random allocation of testing conditions. For the upper trapezius muscle, the main effects were significant for shoulder flexion angle (p < 0.001) and feedback (p = 0.017), as was the interaction effect (p = 0.003). Visual feedback increased activation of the upper trapezius at both 60°

and 90° of shoulder flexion ( Table 2). After Bonferroni correction, however, the effect of visual feedback was significant only at the 60° shoulder flexion angle (p = 0.008). For the lower trapezius muscle, the main effect for shoulder flexion angle was significant (p = 0.001), but neither the main http://www.selleckchem.com/products/DAPT-GSI-IX.html effect for the visual-feedback condition (p = 0.152) nor the interaction effect (p = 0.150) was significant. The data are presented in Table 2. For the serratus anterior muscle, the main effects were significant for shoulder flexion angle (p < 0.001) and feedback

(p < 0.001), as was the interaction effect (p = 0.045). Visual feedback significantly increased activation of serratus anterior at both 60° and 90° of shoulder flexion ( Table 2). After Bonferroni correction, the effect of visual feedback remained significant at both 60° and 90° of shoulder flexion (p < 0.001). Measurement of displacement of the acromial marker in the frontal plane showed that the average movement was superior for all combinations of flexion angle and feedback. The main effects were significant for shoulder flexion angle (p < 0.001) and feedback and (p < 0.001), as was the interaction effect (p = 0.001). Visual feedback significantly increased the superior displacement of the acromion ( Table 3). After Bonferroni correction, the effect of feedback remained significant only at 60° of shoulder flexion (p < 0.001). Measurement of displacement of the acromial marker in the sagittal plane showed that the average movement was anterior with feedback and posterior without feedback. The main effect was significant for the visual feedback (p = 0.000), but neither the main effect for shoulder flexion angle (p = 0.100) nor the interaction (p = 0.268) was significant. After Bonferroni correction, the effect of visual feedback on anterior movement of the acromion during shoulder flexion remained significant at both 60° and 90° of shoulder flexion (p < 0.001).

Il semble donc qu’il faille globaliser l’ensemble des nouveaux an

Il semble donc qu’il faille globaliser l’ensemble des nouveaux anticoagulants oraux (dabigatran, rivaroxaban, apixaban et bientôt edoxaban), pour simplifier selleck products leur gestion péri-opératoire et adopter une seule politique commune. En chirurgie réglée, une interruption des traitements 5 jours avant la procédure semble suffisante au vu de la pharmacocinétique de ces produits. Le dabigatran, dont l’élimination est essentiellement rénale et la demi-vie de 17 heures, n’est (le plus souvent…) plus présent dans la circulation plasmatique au-delà des 4 jours. Pour le rivaroxaban, dont la demi-vie varie entre 7 et 13 heures selon

l’âge et le statut clinique, le délai est un peu plus court. L’apixaban a, quant à lui, une demi-vie de 10 à 15 heures [26]. Cinq jours d’interruption paraissent donc un délai de sécurité suffisant, sauf peut-être chez les patients insuffisants rénaux modérés (clairance de la créatinine entre 30 et 50 mL/min) traités par dabigatran. NVP-AUY922 clinical trial Les patients pourraient être gérés en adoptant une stratégie mimant les recommandations de la Haute Autorité de santé française sur

les AVK [27]. La même stratification pourrait être proposée mettant d’un côté des patients à risque thrombotique élevé qui vont bénéficier d’un relais par HBPM (deux injections sous-cutanées par jour…) et les autres. Il s’agit des patients en arythmie complète avec un score de CHADS ≥ 2 ou des patients traités récemment pour un événement thrombo-embolique veineux. Les patients porteurs d’une valve mécanique sont exclus de cette approche car les NACO ne sont pas autorisés whatever ici. Pour les autres patients, traités pour un risque thrombotique moins important, l’arrêt

simple du traitement anticoagulant oral pendant 5 jours semble suffisant, sans relais par HBPM (figure 1). Enfin, un certain nombre de procédures actuellement réalisées sans interruption des AVK, comme la chirurgie bucco-dentaire ou la plupart des endoscopies digestives, doivent très probablement pouvoir aussi être réalisées sous NACO, ou après une interruption de 24 heures. Le GIHP propose la reprise à dose prophylactique le soir suivant l’intervention uniquement pour la prothèse totale de hanche et de genou (AMM). Dans les autres cas, une HBPM sera utilisée à dose préventive, jusqu’à ce que l’hémostase chirurgicale soit stabilisée et/ou que le cathéter d’anesthésie locorégionale soit enlevé. Puis, le traitement par NACO à dose curative est ensuite repris, le plus souvent à la 72e heure. De nombreuses questions demeurent, dont celle de l’arrivée en urgence d’un patient traité à dose efficace (dose thérapeutique) avec un nouvel anticoagulant oral. Le dabigatran est dialysable ; ce n’est pas le cas du rivaroxaban et pour l’instant aucun antidote n’est disponible.

This is consistent with a prospective

study on the outcom

This is consistent with a prospective

study on the outcomes of 120 community-dwelling women after hip fracture (Williams et al 1994a, Williams et al 1994b). In this study, ZD1839 ic50 mobility recovery continued during the first 14 weeks after fracture with the most rapid change occurring between two and eight weeks. A physiotherapist should have reviewed participants’ mobility over this period, and certainly beyond the first six weeks after discharge. Yet, nearly 94% of participants reported that no review date had been scheduled and, as it currently stands in South Australia, most rehabilitation ceases within six weeks post fracture, which is short of what would appear to be the optimum mobility review period. Some limitations of this study are acknowledged. The study participants were enrolled in a randomised trial and therefore may not have been a representative sample of hip fracture patients. Autophagy signaling pathway inhibitor However, it is likely that we recruited patients with sufficient cognitive ability and social supports to allow participation in a clinical trial. Therefore, our results are likely to underestimate the misuse of walking aids by patients discharged

from hospitals after hip fracture. Further underestimation may have occurred due to the exclusion of non-English speaking people. They are potentially at greater risk of not receiving clear instructions regarding walking aid prescription and use, due to communication barriers between patients and therapists. Another limitation is that the findings around whether goals had been established or if education on walking aid use had been provided relied heavily on recall by the participant. Possibly physiotherapists did put

plans in place and explained to participants how to progress their walking aids, but participants could not recall this having occurred. Regardless, this highlights the need for follow up, because even if participants did receive the information during their admission, this study shows that they are unlikely to retain this information after discharge. Also, it cannot be ignored Farnesyltransferase that half of the observed participants in this study were receiving an additional intense exercise intervention as part of a clinical trial. Although reviewing and progressing the walking aids of individual participants was not the primary aim of the research physiotherapist, it is possible that the physiotherapist was more proactive with the intervention group than the control group in providing advice and education regarding walking aid use. This could have influenced the length of time until a participant changed their walking aid, or the appropriateness of walking aid use. However, this would be expected to have had a positive effect on walking aid use. In conclusion, follow up by physiotherapists of walking aid use in the early recovery phase of hip fracture is limited and walking aid misuse is common in the first six months of recovery.

To generate the final vaccine strain, we deleted lpxL1 and engine

To generate the final vaccine strain, we deleted lpxL1 and engineered the mutant to over-express fHbp v.1, designated ‘Triple KO, OE fHbp’. We also prepared two isogenic group W control strains: one with deleted lpxL1 and gna33, over-expressed fHbp v.1 with the capsule still expressed (‘Double KO, OE fHbp’), and

another with deleted lpxL1, capsule and gna33, but no fHbp over-expression (‘Triple KO’) ( Table 2). SDS–PAGE and Coomassie Blue staining of the proteins revealed a similar protein pattern in the three GMMA preparations. Densitometry indicated that in all three GMMA BMS-354825 cell line preparations, the relative amount of PorA to total protein is 5%. By silver stain, the GMMA contained similar levels of lipooligosaccharide. By capture ELISA, with recombinant fHbp as standard, approximately 3% of the total protein in Dabrafenib nmr GMMA from the Triple KO, OE fHbp was fHbp, and by Western blot, the two GMMA over-expressing fHbp had similar fHbp levels. To assess the endotoxic activity of the GMMA, we measured the release of

IL-6 by human PBMC after stimulation with different concentrations of GMMA from the Triple KO, OE fHbp mutant and the parent serogroup W wild type strain (Fig. 1C). Approximately 50-fold higher concentrations of GMMA from the mutant strain were required to stimulate the release of 200 pg/mL IL-6, confirming the decrease in endotoxic activity. We measured the ability of the GMMA to stimulate human TLR-4 in transfected HEK293 cells (Fig. 1D). Low concentrations of GMMA from the wild type bacteria stimulated TLR-4, as measured by increased NF-κB expression. Approximately 1000-fold higher concentrations of GMMA from the Triple KO, OE fHbp mutant were required for equivalent TLR-4 stimulation. These results are consistent with a strongly decreased ability of the LOS in GMMA from the serogroup W mutant to activate TLR-4 compared with GMMA from the non-detoxified parent wild type strain. We measured anti-fHbp v.1 antibody responses in individual serum samples by ELISA. GMMA from all mutants with

over-expressed fHbp elicited high anti-fHbp antibody responses, even at Sodium butyrate the lowest dose of 0.2 μg (Fig. 2). 5 μg Triple KO, OE fHbp GMMA induced significantly higher geometric mean titres than 5 μg Double KO, OE fHbp GMMA (P = 0.03) or 5 μg of recombinant fHbp v.1 (P < 0.001). GMMA from the Triple KO mutant without fHbp over-expression induced no measurable anti-fHbp antibody responses. The three serogroup W test strains were isolated in Ghana, Mali and Burkina Faso and expressed PorA subtype P1.5,2, which is identical to that expressed by the GMMA vaccine strains. Strain BF2/11 expressed fHbp v.1 (ID9) and the two other strains expressed fHbp v.2 (ID23). The seven group A strains tested were collected in Ghana, Burkina Faso, Sudan and Mali. They expressed a heterologous PorA compared to that in the GMMA, and fHbp v.1 (ID5).

56 μg/mL on both the cancer cell lines ( Tables 1 and 2) Figs 1

56 μg/mL on both the cancer cell lines ( Tables 1 and 2). Figs. 1 and 2 depict the morphological changes in A549 and MCF-7 cells treated with methanolic extracts of B. hispida and M. dioica, indicating the formation of spherical shaped cells which is the onset of apoptosis occurring in the concentration dependent manner. As the cell’s intrinsic cell death program, apoptosis plays a key role in growth control of cells and tissue homeostasis. Therefore, the induction and recovery of the apoptotic response in tumor cells are relevant steps in anticancer treatment. selleck The anticancer potential

of Rubiaceae plant species has been recorded, which includes the cell growth inhibiting effects of methanolic leaf extract of Oldenlandia diffusa (Rubiaceae), on different find more cancer cell lines. 13 The methanol extract of the flowers of Ixora coccinea L. (Rubiaceae), contain ursolic acid, which is known to posses antitumor activity. 14 The antitumor activity of methanol extract of aerial parts of Cucurbita maxima (Cucurbitacae), 15 and Momordica cymbalaria 16 was identified on Ehrlich Ascites Carcinoma (EAC) models in mice. Kiran et al reported that the anticancer effects of methanol extract of leaves of Argemone mexicana exhibited the inhibition of cell growth at the IC50 value of 1.35 μg/mL for MCF-7 cells. 17 According to the above mentioned discussions, it can be stated that methanolic extracts of plants may contain

potential compounds or active principles which can act as

effective sources of anticancer agents. The results of this study indicate that the methanolic seed 17-DMAG (Alvespimycin) HCl extract of B. hispida showed anticancer activity by causing 50% inhibition of A549 cells at IC50 value of 3.125 μg/mL and MCF-7 cells at IC50 value of 1.56 μg/mL. Inhibitory action was also observed with the treatment of methanolic seed extracts of M. dioica on A549 cells at the IC50 value of 12.5 μg/mL and on MCF-7 cells at the IC50 value of 3.125 μg/mL. In essence, the present work revealed that B. hispida and M. dioica, contains some important chemical constituents extracted using methanol as solvent, that can be used further in the management of cancer treatment. All authors have none to declare. “
“Diabetes has been estimated to affect 177 million people worldwide, and this figure is estimated to increase 300 million by 2025.1 Type 2 diabetes mellitus (non-insulin-dependent diabetes) is one of the most common chronic diseases and is associated with co-morbidities such as obesity, hypertension, hyperlipidemia and cardiovascular diseases.2 Currently a variety of therapeutic drugs are available for management of type 2 diabetes, such as acarbose, migitol and voglibose known to inhibit a wide range of glycosidases. But these drugs have certain adverse effects such as hyperglycemia at higher doses, liver problems, lactic acidosis, and diarrhea.

Such protocols can be adapted to consider developing epidemiologi

Such protocols can be adapted to consider developing epidemiological information and in consultation with advisory groups, thus limiting some of the crucial decision-making needed in the midst of an emergency. Vaccine procurement and access in the Americas was differential and not equitable. The first LAC countries to have access to pandemic vaccine were those with pre-existing agreements with manufacturers. Regional or sub-regional efforts should be undertaken to enhance and extend current transfer of technology agreements for vaccine production

in LAC. PAHO’s RF played a key role in providing countries and territories, especially small ones, RAD001 manufacturer access to the vaccine. The vaccine donation coordinated through WHO was an international diplomatic effort aiming to provide vaccine Proteasome inhibition to those countries with less economic resources. Unfortunately, the donation process proved to be lengthy, resulting in recipient countries being the last to receive vaccine in LAC. Efforts to streamline future donation processes are necessary to ensure the timeliness and equity of such endeavors. Many LAC countries successfully implemented pandemic vaccination

campaigns, making use of the current infrastructure of the national immunization programs. They reached, on average, 99% of their pre-defined high risk target populations. However, countries had to face multiple technical and logistical challenges, including multiple vaccine presentations, vaccine with and without adjuvant, multiple vaccine shipments due to ongoing production, and non-traditional target groups. Clear guidelines and training workshops conducted prior to vaccine arrival were critical for capacity building of health-care workers to help them manage said challenges. Pregnant

women had the lowest pandemic influenza (H1N1) vaccine coverage. In some countries health care professionals were reluctant to recommend the vaccine. This issue highlights the need to enhance health-care worker training, increase the participation of scientific societies of obstetricians and gynecologists, and strengthen social communication regarding the benefits of influenza vaccination for pregnant of women. These lessons can also be applied to annual seasonal influenza vaccination. Given the magnitude of vaccination activities in LAC and the commitment of countries to such an effort, it is important to assess the impact of this investment in the reduction of influenza disease burden. Estimations of the impact of vaccination are underway in selected countries and have presented a series of challenges, including the absence of serosurveys conducted prior to vaccine introduction, and a lack of surveillance data stratified by vaccine target groups.