The resulting plasmids were designated as WT1 − 17AA/− KTS −, + 17AA/− KTS −, − 17AA/+ KTS −, or + 17AA/+ KTS-pHR-SIN-CSGW dlNotI. We deleted the eGFP sequence in pHR-SIN-CSGW dlNotI and used this modified vector as a control vector. Preparation of infectious particles was performed according to established protocols [29]. click here All of the procedures involving animals in this study were approved by the animal care committee of Yamagata University in accordance with institutional and Japanese government guidelines for animal experiments. WT1 splice variants (− 17AA/− KTS, + 17AA/− KTS, − 17AA/+ KTS, and + 17AA/+ KTS) or control lentivirus vectors were co-transfected with packaging plasmids (pVSV-G

and pGag/pol) into 293 T cells to generate lentiviral particles. These particles were then used to transduce SKOV3ip1 cells, and cells stably expressing the vectors were selected with 1 μg/mL puromycin. SKOV3ip1 cells (2 × 106) expressing WT1 variants (− 17AA/− KTS [n = 13], + 17AA/− KTS [n = 13], − 17AA/+ KTS [n = 8], or + 17AA/+ KTS [n = 10]) or control vector (n = 8) were suspended in 250 μL PBS and inoculated by intraperitoneal injection (i.p.) into Roxadustat concentration 5- to 7-week-old female BALB/CA nu/nu mice. Abdominal circumference and body weight were measured twice weekly. Mice injected with WT1 − 17AA/− KTS-expressing cells were euthanized with CO2 after 36 days and mice injected with cells expressing

control vector or the other variants were euthanized after 40 days to assess tumorigenecity by measuring volume of ascites, extent of dissemination, and weight of tumors. We used data from mice that were Protein kinase N1 euthanized precisely after 36 or 40 days. In a second experiment, 2 × 106 SKOV3ip1 cells expressing the control vector or each WT1 variant were injected i.p. into 5- to 7-week-old female BALB/CA nu/nu mice (n = 30, with 6 mice per group of cells), and survival was evaluated from the first

day of inoculation until death. In a third experiment, SKOV3ip1 cells (2 × 106) expressing WT1 − 17AA/− KTS were implanted by i.p. into 5- to 7-week-old nu/nu nude mice (n = 10). After two week after inoculation, one group of mice (n = 5) was treated i.p with PBS twice weekly for 3 weeks. A second group of mice (n = 5) was treated ip with bevacizumab (5 mg/kg) twice weekly for 3 weeks. Bevacizumab was kindly provided by Chugai Pharmaceutical Co., Ltd. (Tokyo, Japan). Bevacizumab were diluted in 200 μl of PBS. Abdominal circumference and body weight were measured once a week. At 3 weeks after initiating treatment, mice were euthanized with CO2 to assess antitumor efficacy of bevacizumab by measuring volume of ascites and weight of tumors. Briefly, tumors were homogenized in RLT buffer. Total RNA was isolated and purified with an RNeasy− Mini kit (Qiagen, Valencia, CA, USA). cDNA was generated form 0.4 μg RNA using a QuantiTect Reverse Transcription kit (Invitrogen). cDNA (1.

prolixus and T. brasiliensis remains puzzling. One Trametinib price possibility for this contradiction might be the differing phylogenetic origin and

biology of these two triatomine species and thus divergent gene expression and physiology ( Araújo et al., 2009). As we showed in the present study, several cathepsin L isoforms are expressed in the triatomine midgut. It is also possible that other isoforms assume the role of this specific R. prolixus cathepsin L, but are not detectable by a highly specific methodology like RT-PCR. However, since we analyzed the cathepsin L transcript abundance in a more detailed way – using more tissues – the cathepsin L expression pattern became clearer. The transcript abundance pattern indicates a major role of the respective enzymes predominantly in the small intestine of fifth instar nymphs and adult insects. Intestinal pH is one important physiological parameter which affects the efficiency of digestive enzymes (Terra et al., 1996). Activity maxima of proteolytic enzymes, evaluated in various studies, emphasize the acid character of

the small intestine content in triatomines (Houseman and Downe, 1980, Houseman and Downe, 1981, Houseman and Downe, 1982 and Houseman and Downe, 1983). Using a microelectrode, the pH measured in the stomach of T. brasiliensis has been between 7.02 and 7.16 ( Barros et al., 2009). However, mixing contents of different midgut regions – e.g. anterior and posterior midgut or ecto- and endo-peritrophic (extra cellular membrane layer in Hemiptera) regions – with contrasting pH values will certainly give inaccurate results ( Terra and Ferreira, 1994). Thus determination

of http://www.selleckchem.com/products/VX-809.html the pH in the whole midgut might reflect the intestinal conditions more precise. The ingested blood surely contributes to the neutral or rather slightly alkaline environment in the stomach, PD184352 (CI-1040) but also guts of non-fed bugs show a pH within the range of 7.0. It remains unclear whether or not cathepsin L is secreted into the lumen of the stomach because, (i) at the neutral pH value present in this midgut region their activity would be very low and (ii) consequently also low propeptide cleavage and enzyme activation by autocatalysis will occur in this environment. Hence only the small intestine lumen with its acid pH offers proper conditions for reasonable cathepsin activity. So far, intestinal proteolytic activities of triatomines have been analyzed by photometric assays. By using specific substrates (e.g. BAPNA, BANA, LPNA and Z-Phe-Arg-pNA), the luminal activity of cathepsin B, D and L, carboxypeptidase A and B and an aminopeptidase has been shown (Houseman and Downe, 1981, Houseman and Downe, 1982, Houseman and Downe, 1983, Kollien et al., 2004 and Borges et al., 2006). Using a biotinyl affinity assay, several putative cysteine proteinases in the range of 30–35 kDa has been shown in the small intestine of T. infestans at 5 daf ( Kollien et al., 2004).

“
“Dietary composition impacts myocardial structure and function. Intake of a “Western (WES)” diet rich in saturated fatty acids and simple carbohydrates is associated with left ventricular hypertrophy (LVH) and diastolic dysfunction [1], [2] and [3]. In contrast, consumption of n-3 polyunsaturated

fatty acids (PUFA) is associated with antihypertrophic effects [4], [5], [6] and [7]. We were therefore surprised to observe that rats fed a WES diet supplemented with docosahexaenoic acid (DHA) for 3 months had similar thickening of the cranial left ventricular (LV) wall compared with rats fed a WES diet alone and had increased caudal LV wall thickness compared with control (CON) animals [3]. These findings led us to check details consider whether the underlying genotype of the myocardial SD-208 order tissue differed despite a similar gross phenotype, that is, whether WES diet consumption promoted a pathologic or maladaptive gene expression profile, whereas DHA treatment was associated with physiologic or adaptive gene expression. Transcriptome profiling in rats has distinguished unique expression patterns in physiologic (adaptive) and pathologic (maladaptive) myocardial hypertrophy, specifically in relation to apoptosis, carbohydrate

metabolism, and protein synthesis [8]. In addition, distinct myocardial expression patterns are associated with diet in mice [9] and specifically with n-3 PUFA, evident in a study of neonatal rat cardiomyocytes [10]. Incubation of these cells with n-3 PUFA revealed differential expression of genes associated with lipid handling, inflammation, cell survival and

proliferation, extracellular matrix remodeling, calcium handling, and oxidative stress [10]. Effects of one-time oral administration of single fatty acids on the myocardial transcriptome in vivo have been documented [11]. To further develop our knowledge using a system relevant to humans, we examined outcomes in response to prolonged oral intake of a combination of fatty acids reflecting that consumed by people, using a normal (without genetic aberrancy or induced pathology) rat model. Our objective was to determine whether long-term DHA supplementation of a WES diet alters gene expression in the rat left ventricle Rutecarpine and whether the expression patterns reflect a physiologic or pathologic response. To answer this question, microarray transcriptome profiling was used to uncover changes in gene expression associated with dietary treatments, followed by quantitative real-time polymerase chain reaction (qRT-PCR), and immunoblotting to validate and further pursue relevant gene pathways. We hypothesized that WES diet consumption would be associated with a pathologic or maladaptive gene expression profile, whereas DHA treatment would favor a physiologic or adaptive expression pattern.

Moving beyond the quantitation of information, key qualitative questions remain about how ‘meaning’ is transferred along with information. This is not merely an abstract question; synthetic biology can engineer reliable information transfer, but how would such systems encode or process higher order meaning, such as the difference between to ‘I must’ and ‘I want to’? Simple IF-THEN logic does not suffice. To harness essential features of biology, synthetic biologists

Cabozantinib chemical structure somehow need to wire components to encode choice and reward, perhaps by including feedbacks in system resource allocation. We still do not know how to engineer higher order meaning, such as desire or fear. While information theory clearly has a part to play in increasing our engineering capability, we also need to develop a functional philosophy of meaning. Papers of particular interest, published within the period of review, have been highlighted as: • of special interest DB and VRS are both Akt inhibitor funded by La Caixa PhD Fellowships. MI is supported by EC FP7-610730 EVOPROG, and Wellcome Trust UK New Investigator Award WT102944.

We thank Jesper Ferkinghoff-Borg for providing us with original images of information channels inside a single protein. “
“The IC50 value of 3 has been reported as 18(5) μM in MCF-7. It actually is 185(5) μM for MCF-7 and hence the corrected Table 3 is as follows: “
“Indazoles are rare in nature, and so far only three natural products based on an indazole ring have been isolated [1]. These are the indazole alkaloids nigellicine [2], nigeglanine [3], and nigellidine [4]. The total syntheses of nigellicine and

nigeglanine are also well documented [5] and [6]. The indazole ring system is of much current interest as partial structure of a large number of biologically active compounds. Different aspects of pharmaceutical and other useful applications of indazoles Histamine H2 receptor have been reviewed [7] and [8]. Some substituted indazoles exhibit relevant biological properties for development as anticancer drugs [9], [10], [11], [12], [13], [14] and [15]. One of the tetrasubstituted indazoles, namely, CI-958, entered clinical trials for prostate cancer treatment about a decade ago [16]. From the unsubstituted indazole derivatives the most prominent example is the ruthenium(III) compound (H2ind)[trans-RuIIICl4(Hind)2] (KP1019, Hind = 1H-indazole), which is now in clinical trials as an anticancer agent against metastatic solid tumors [17] and [18]. Of potential interest are also complexes closely related to (H2im)[trans-RuIIICl4(DMSO)(Him)] (NAMI-A, Him = imidazole) [19], an investigational drug which is currently evaluated in a clinical phase II trial for its capacity of inhibiting the process of metastasis, namely (H2ind)[trans-RuIIICl4(DMSO)(Hind)] [20] and its osmium counterpart [21].

Journal of Coastal Research 21, 421–429. should TSA HDAC have been presented as Walton Jr., T.L., 2005. Short term storm surge forecasting. Journal of Coastal Research 21 (3) 421–429. Further, the

corresponding citations of Todd and Walton (2005) in the text should have been cited as Walton (2005). “
“Winter cooling and sea ice formation forms large amounts of brine-enriched shelf water over the vast shelves in the Arctic Ocean. Plumes of dense shelf water eventually spill over the continental shelf edge and flow down the slopes as dense water cascades (see e.g. Ivanov et al., 2004, for an overview of known cascading locations in the Arctic and other oceans). During their descent the cascading plumes entrain the ambient water, lose their initial density gradient and eventually this website disperse laterally into the ambient stratification (e.g. Aagaard et al., 1985, Jungclaus et al., 1995 and Shapiro et al., 2003). Dense water formation is particularly intense in coastal polynyas, which are estimated to produce a total of 0.7–1.2 Sv ( 1Sv≡106m3s-1) of dense water

over the entire Arctic Ocean (Cavalieri and Martin, 1994), making this process of deep water formation comparable to open ocean convection in the Greenland Sea (Smethie et al., 1986). The dense waters formed on the shelves thus significantly influence the heat and salt balance of the entire Arctic Ocean (Aagaard et al., 1985). Cascading also contributes to

the maintenance of the cold halocline layer (Aagaard et al., 1981) and the replenishment of intermediate and deep Arctic 4-Aminobutyrate aminotransferase waters (Rudels and Quadfasel, 1991 and Rudels et al., 1994). A well-known site of dense water formation and subsequent cascading is the Storfjorden, located between 76°30”–78°30” N and 17°–22° W in the south of the Svalbard archipelago (Fig. 1). Each winter, intense sea ice production and brine-rejection in a recurring latent-heat polynya in Storfjorden forms significant amounts of dense water (Schauer, 1995, Haarpaintner et al., 2001 and Skogseth et al., 2005b) which eventually spill over the sill located at approx. 77°N and 19°E at a depth of 115 m (Skogseth et al., 2005a and Geyer et al., 2009). Near the sill the overflow plume encounters the relatively fresh and cold East Spitsbergen Water (ESW) which mainly reduces its salinity (Fer et al., 2003). The flow is then channelled through the Storfjordrenna on a westwards path, before it bends northwards to follow the continental slope of western Spitsbergen (see Fig. 1, Quadfasel et al., 1988, Fer and Ådlandsvik, 2008 and Akimova et al., 2011). The lighter fractions of the overflow water remain within the depth range of the Atlantic Water (approx.

“
“Functional constipation is a common gastrointestinal problem in children. The estimated worldwide prevalence varies from 1% to 30% [1] and [2]. Currently, the diagnosis of functional constipation is based on the Rome III criteria and includes two or more of the following: ≤2 defecations in the toilet per week; at least one episode of fecal incontinence per week; history of retentive posturing or excessive volitional stool retention; history of painful or

hard bowel movements; presence of a large fecal mass in the rectum; and a history of large-diameter stools which may obstruct the toilet [3]. The criteria are fulfilled selleck chemical when their defining symptoms appear at least once per week for at least 2 months prior to diagnosis [3]. Evidence-based guidelines from the North American Society of Pediatric Gastroenterology, Hepatology and Nutrition (NASPGHAN) [4], as well as the National Institute for Health and Clinical Excellence (NICE) guidelines [5], consistently

recommend disimpaction, if present, followed by a maintenance therapy. Available therapeutic measures include toilet training and the use of oral osmotic laxatives (e.g., lactulose, polyethylene glycol), stimulant laxatives (e.g., bisacodyl), or mineral oil [4], [5] and [6]. However, none of these measures offers long-lasting effects, hence, interest in alternative therapies. Previously, we evaluated the effect of gut microbiota modification with prebiotics or probiotics in children with functional constipation in 2 randomized controlled trials [7] and [8]. The rationale for the use of prebiotics/probiotics Epacadostat supplier in the treatment of functional constipation was based on data demonstrating differences in the intestinal microbiota between healthy individuals and patients with chronic constipation [9] and [10]. In these studies, the rate of treatment success ranged from 57% [8] to 67% [9], but there was no difference between the groups in any of the studies. Constipation unfavorably influences the quality of life of affected children [11] and [12]. While the goal of treatment http://www.selleck.co.jp/products/AP24534.html of functional

constipation is to restore a regular defecation pattern and to prevent relapses, the persistence of symptoms of constipation was reported in 30–52% of children followed up for at least 5 years [13] and [14]. This indicates that functional constipation is not a transient, mild disorder. Data from Poland are limited. The aim of the current study was to assess long-term outcomes in children with functional constipation who had participated in those 2 previous trials [8] and [9]. The current trial was a follow-up study of children who had participated in 2 previously published, randomized controlled trials carried out at our center. The designs of these studies have been described elsewhere [8] and [9]. Briefly, in the first trial (n = 80) [8], children aged 3–16 years with functional constipation according to the Rome III criteria were randomly assigned to receive glucomannan (GNN), 2.

S1). The PCR products for each variable region were pooled according to the natural distribution as described on V-Base. The light chain variable regions were cloned first using restriction digest with SfiI and AvrII for Vλ and SfiI and BsiWI for Vκ and transformed into electrocompetent TG1 cells (48 μg DNA in 48 200 μL Trametinib transformations for Vκ and 65 μg DNA in 65 200 μL transformations for Vλ). Transformations were spread on 2xYT medium with 2% glucose and 100 μg/mL carbenicillin, which were incubated overnight at 30 °C. The following morning the bacteria were scraped from the plates, combined and plasmid DNA purified

with the GenElute™ HP Maxiprep Kit (Sigma-Aldrich). The resulting DNA was prepared for cloning VH with NcoI-HF and NheI-HF. The ligated DNA was cleaned with the Wizard® SV Gel and PCR Clean-up system (Promega) and transformed into electrocompetent TG1 cells (66 μg DNA in 66 200 μL transformations for Vκ and 100 μg DNA in 100 200 μL transformations for Vλ). Transformations were spread on 2xYT medium Epacadostat solubility dmso with 2% glucose and 100 μg/mL carbenicillin, which were incubated overnight at 30 °C. The following morning the bacteria were scraped from the plates, combined, and stored in 15% glycerol 2xYT at − 80 °C. The scFv library was constructed similarly to the above described Fab library with the following changes. Primer sequences are listed in Table S3 and Table S4. cDNA from 20 PBMC samples, 8 bone marrow

samples, 1 lymph node sample, and 1 spleen sample Fenbendazole were used. The reverse secondary PCR primers for VH and forward secondary primers for Vκ and Vλ had complementary extensions for an AST(G4S)3 linker and the forward secondary PCR

primers for VH and reverse secondary primers for Vκ and Vλ had sequences to add flanking SfiI restriction sites. A tertiary PCR step was then done to assemble the full length scFv fragment, which was next cloned into pXHMV-scFv ( Fig. S1) using the SfiI sites. The ligated DNA was transformed into electrocompetent TG1 cells (147 μg DNA in 120 200 μL transformations for Vκ and 44 μg DNA in 40 200 μL transformations for Vλ). Transformations were spread on 2xYT medium with 2% glucose and 100 μg/mL carbenicillin, which were incubated overnight at 30 °C. The following morning the bacteria were scraped from the plates, combined, and stored in 15% glycerol 2xYT at − 80 °C. Both XFab1 and XscFv2 phage libraries were rescued using a modification of the standard protocol (Marks et al., 1991). XFab1 was rescued in four batches (two for XFab1λ and two for XFab1κ) each starting with 5-fold more bacteria than the sub-library size. XscFv2 was rescued in five batches (two for XscFv2λ and three for XscFv2κ) with XscFv2λ starting 5-fold more bacteria than the sub-library size and XscFv2κ starting with 3.33-fold more bacteria than the sub-library size. For all rescue batches, cultures were seeded at a starting density of 0.

We asked them to respond as quickly and accurately as possible. Participants received feedback on accuracy on each trial (750 msec). The aim of Experiment 2 was to examine the impact of spatial location in synaesthetic experience. We tested this by manipulating the on-screen position of targets. The spatial congruency was defined by where the target was positioned on the computer screen in relation to where synaesthetes positioned their drawing on the screen or paper. For each

synaesthete, we used the same set of four sound–image pairs as those in Experiment 1 such that the images were manifestly distinct from each other in colour, shape, and location. The design, procedure, and instructions of Experiment 2 selleck compound were identical to Experiment 1, with the exception that we manipulated the on-screen position of targets, while keeping one of the other visual features constant. In the colour task, the image colour and on-screen location were either Selleckchem ABT199 congruent or incongruent with the synaesthetic colour and location while

the synaesthetic shape induced by the sound was always consistent with the image shape. Conversely, in the shape task, shape and location were independently manipulated while synaesthetic colour was always consistent with image colour. As a result, two different versions of the stimuli were used in the colour and shape tasks. There were four conditions for each task: (1) both features congruent; (2) location incongruent; (3) colour or shape incongruent (in the colour / shape task, respectively); and (4) both Pregnenolone features incongruent. Although the reported experiences initially seem idiosyncratic and variable across synaesthetes, there is a systematic relationship between auditory pitch and visual features: in all seven synaesthetes, high-pitched sounds induce visual experiences that are brighter in colour, smaller in size, and higher in space, relative to low-pitched sounds. Fig. 3 illustrates the pattern of the synaesthetic experiences from two representative participants. Such a pattern bears similarity to previous research on

the way non-synaesthetes map auditory pitch to visual features (Spence, 2011), and is also consistent with Ward et al. (2006) who reported similarities between synaesthetes and non-synaesthetes in auditory–visual mappings. To quantify the phenomenological relationship between auditory pitch and the size, brightness, and location of synaesthetic objects, we performed correlation analyses: for each of the seven synaesthetes, we calculated the size (number of pixels) of the synaesthetic object and brightness of the selected colour (in Hue-Saturation-Brightness colour coordinates, ranging from 0 to 100) using Photoshop (hand-drawings were scanned and converted into JPG files). If multiple colours were present in an image, we used the colour that occupied the most area.

Typhimurium ( MacLennan et al., 2010) and this warrants further investigation. Despite its probable importance, little is known about the natural immune response to LPS. The capacity to purify LPS-specific antibodies would, for example, be useful in analysing V region usage. Purification of Salmonella OAg-antibodies from polyclonal sera would allow further characterisation of both the functionality and specificity of these antibodies. This would facilitate the ongoing investigation of their potential protective and blocking effects in individuals immunised with OAg-based vaccines and in HIV-infected African adults. Monoclonal and polyclonal

antibodies are conventionally purified by affinity chromatography (Cuatrecasas, 1970, Jack, 1994 and Huse et al., 2002), using the highly-specific nature of the interaction between antigen and antibody. EPZ015666 in vitro The antigen is covalently attached to a solid support under conditions that retain antibody-binding capacity. Subsequently, when serum is passed over the antigen-bound column, only those molecules with specific affinity for the antigen are bound. After washing, the bound antibodies are eluted, thereby purifying them from the original sample. Although this method for recovering active antibodies is potentially selective, rapid and simple, allowing antibody purification

in a single chromatographic step, the recovery is typically low (Casey et al., 1995 and Cuatrecasas

and Anfinsen, 1971). C646 purchase Optimised conditions need to be determined to permit efficient purification of the desired antibodies without altering their native structure (Narhi et al., 1997a). Salmonella LPS consists of lipid A linked to the 3-deoxy-D-manno-octulosonic acid (KDO) terminus of the conserved core region, which in turn is linked to the serovar-specific OAg chain. The OAg chain is the immunodominant portion of the molecule and extends as a repeating aminophylline polymer from the end of the core region ( Whitfield et al., 2003). In S. Typhimurium, the OAg repeat (O:4,5) consists of a trisaccharide backbone, with a branch of abequose, usually O-acetylated on C-2, which confers serogroup specificity (factor 4,5) ( Fig. 1A) ( Hellerqvicst et al., 1969). LPS detoxification is usually performed by acetic acid hydrolysis or by hydrazinolysis (Konadu et al., 1996), with the former commonly preferred as it retains the O-acetyl groups along the OAg chain. Acid hydrolysis cleaves the labile linkage between Lipid A and KDO leaving the OAg chain attached to the core region (Fig. 1A). Many approaches have been used to bind LPS or detoxified OAg from various bacteria to resins for use in affinity purification and, despite the high toxicity, CNBr-activated resin has been the most commonly employed (Stiller and Nielsen, 1983 and Rodahl and Maeland, 1984).

Additional desirable features include the ability to engineer and deliver genetic adjuvants in tandem or parallel with the antigen, the potential to deliver multiple antigen genes in one construct or within other constructs that encode adjuvanting protein(s), and the ability to induce both cellular and humoral immune responses. Despite promising data in pre-clinical testing, DNA vaccine candidates have shown only limited success in clinical settings so far. One of the current

drawbacks of DNA learn more vaccines is the inefficiency of conventional delivery methods for the plasmid DNA; however, emerging proprietary particle-mediated delivery technology or electroporation technology seeks to check details improve this situation. With the electroporation method, brief electrical pulses are applied at the site of immunisation which causes a transient disruption of cell membranes. This results in an enhancement in uptake of the DNA vaccine between 10–100-fold. Examples of DNA candidate vaccines in clinical development are presented

in Table 6.5. Dendritic cell (DC) vaccines typically use monocytes harvested from the blood (in most cases from the individual who will receive the vaccine) to produce immature DCs in vitro. The monocytes are antigen-loaded and treated to induce their maturation into APCs and infused back into the

patient. The first Food and Drug Administration (FDA)-approved DC vaccine, designed for the treatment of prostate cancer, was licensed in 2010 (Sipuleucel-T); examples of other targets for DC vaccine therapy are presented in Table 6.6. DC vaccines offer an individualised approach to therapeutic vaccine development, but represent a specialised method of vaccination that is currently limited to aggressive cancers, and the treatment of serious, intractable infections. DC vaccines hold great Megestrol Acetate promise for the treatment of cancer, HIV and other chronic infections. Utilising the patient’s own DCs, this is truly an individualised biomedical intervention. A comparison between the strengths and weaknesses of selected new vaccine platforms is presented in Table 6.7. Developing administration techniques that place the vaccine directly at the site(s) where pathogens are most likely to initiate an infection (eg mucosal or respiratory sites) is likely to improve vaccine efficacy and safety. Traditional methods of vaccine administration can potentially pose a number of limitations with respect to reactogenicity, immunogenicity, convenience, efficacy, safety and cost-effectiveness.