We thank Mari Koivisto, Department of Biostatistics, University o

We thank Mari Koivisto, Department of Biostatistics, University of Turku, Finland for help with the statistical analyses. Conflict of interest statement: AK has participated as a member in advisory boards of Pfizer, GlaxoSmithKline and Novartis and received honorarium from these. She has acted as a consultant to Crucell on vaccination immunology and been reimbursed for giving lectures by GPCR Compound Library concentration Crucell, GSK and Bayer. SHP and JMK declare no conflicts of interest. “
“Meningitidis and sepsis caused by serogroup B meningococcus are two severe diseases that

continue to cause significant mortality [1] and [2]. Five major pathogenic serogroups have been identified on the basis of the chemical composition of the bacterial capsule (A, B, C, Y and W135) [3], [4] and [5]. selleck chemical However, the capsular vaccine approach is not suitable for strains of serogroup B since that polysaccharide capsule

has a structural homology to human embryonic neural tissue [6]. Thus, outer membrane proteins or outer membrane vesicles (OMV)-based vaccines were tested extensively in clinical trials [7]. An alternative approach to vaccine development is based on surface-exposed proteins contained in outer membrane vesicles [4], [8] and [9]. OMV are released from the outer membrane of Gram negative bacteria. They consist of a phospholipid (PL) bilayer containing outer membrane proteins, lipopolysacchharide

(LPS) and periplasmic constituents [10]. These vesicles are made up of five major proteins. Besides, there is the protein NadA and, depending on the conditions of cultivation, the iron regulated proteins (IRP) [11], [12] and [13]. Furthermore, it is worth mentioning that OMV are also employed as carriers of polysaccharides in conjugated vaccines against Haemophilus influenzae and in vaccines against pneumonia [14] and [15]. A common antimeningococcal vaccine project against meningitis B and C had proposed a vaccine containing outer membrane vesicles (OMV) from Neisseria meningitidis B expressing iron regulated proteins (IRP) from a strain with high incidence in Brazil (N 44/89). The lipooligosaccharide (LOS endotoxin) of OMV is high others toxic. However residual LOS amounts are needed to maintain vesicle structure and adjuvate the immune response. Many studies have been carried out previously on other aspects of vaccine development, such as: the production process of N. meningitidis C [16], [17] and [18]; the evaluation of the importance of a second serogroup B strain as vaccine component [19]; the obtainment of vesicles with appropriate characteristics (with IRP expression and with low level of LOS) [20] and [21]; and the conjugation process of N. meningitidis C polysaccharide with N. meningitidis B OMV [22] and [23]. The objective of this study was to investigate the N.

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Sílvio Sanches Veiga for the Alexa 488 anti-mouse


Sílvio Sanches Veiga for the Alexa 488 anti-mouse

immunoglobulin G. This work was supported by MEK inhibitor Fapemig and Fundação Araucária/PPSUS (11403/192). Conflict of interest statement: The authors declare that this research was conducted in the absence of any commercial relationship that could create a potential conflict of interest. “
“Many earlier studies have demonstrated that rotaviruses, like any other enteric viruses are shed in stools and primarily transmitted through fecal-oral route, person-to-person contact and fomites [1] and [2]. There has been evidence that rotaviruses may also be transmitted to individuals through respiratory droplets [2], [3] and [4]. The human rotavirus vaccine strain, HRV mimics natural rotavirus infection, replicates in the intestine of the vaccinated infants and provides protection against future rotavirus infections [5]. Studies with the human rotavirus vaccine have demonstrated that the vaccine virus is shed in the stools of vaccinated infants, with the peak shedding observed on Day 7 after first dose (76–80% of infants after Dose 1 and 18–29% of infants after Dose 2) [6]. Due

to the shedding of infectious vaccine virus in stools, there is a theoretical possibility for vaccine virus to be transmitted to unvaccinated or naive infants—a process similar to that observed in natural wild-type rotavirus infection ISRIB research buy [7]. Such transmissions are possibly expected from any live attenuated vaccines such as oral polio vaccine [8]. The phenomenon of transmission of the rotavirus vaccine strain to unvaccinated individuals raises questions about

the safety of the vaccine and the possibility of conferring indirect protection particularly in developing country settings where the vaccine coverage might be incomplete as compared to the developed countries [9]. The current study was the first of its kind that explored the possibility of horizontal transmission of the HRV rotavirus vaccine strain from one twin who received HRV vaccine to the other twin who received placebo Vasopressin Receptor living under the same household. The immunogenicity and safety of the rotavirus vaccine in transmission cases was also assessed. This phase IIIb, randomized (1:1), placebo-controlled, double-blind study conducted at one urban site in Santo Domingo, Dominican Republic (106260/NCT00396630). Baseline data from all major pediatric hospitals and nurseries was obtained in advance. Parents were informed of the study by presentations at maternity centers, distribution of brochures in health centers and by providing information to pregnant women and new parents visiting maternity centers and vaccination sites. Pairs of healthy twins living in the same household, aged 6–14 weeks at the time of enrolment, born after a gestational period of ≥32 weeks attending local primary healthcare centers, were referred to the site and recruited by the participating physicians.

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, 2013) Comprehensive smoke-free policies have high levels of pu

, 2013). Comprehensive smoke-free policies have high levels of public support and have been associated with substantial health benefits (Fong et al., 2006, International Agency for Research on Cancer, 2009 and Tang et al., 2003). These include reduced tobacco consumption and increased quit attempts, the virtual elimination of SHS from workplaces, lower hospital admission rates for myocardial infarction and stroke, lower admissions BGJ398 for acute respiratory illness in both children and adults (Millett et al.,

2013 and Tan and Glantz, 2012), and lower rates of small for gestational age births (Kabir et al., 2013). However, these health benefits are not equitably distributed as only 16% of the world’s population are covered by comprehensive smoke-free policies (World Health Organization, 2013b). Research evidence suggests that smoke-free workplace policies may change social norms about exposing others to SHS in the home (Berg et al., 2012, Cheng et al., 2011, Fong et al., 2006 and St. Claire et al., 2012). These findings indicate that early concerns that smoke-free workplace policies would lead to behavioural compensation

through an increase in smoking at home have not materialized; rather, results from richer countries ( Berg et al., 2012, Cheng et al., 2011 and St. Claire et al., 2012) and India ( Lee et al., 2013) have consistently found that people employed in a smoke-free workplace are more likely to live in a smoke-free home. Replication of this finding in other LMICs would indicate that implementation of Protein Tyrosine Kinase inhibitor before smoke-free policies in these settings will likely result in substantial reductions in tobacco related harm

globally. This study examines whether there is an association between being employed in a smoke-free workplace and living in a smoke-free home in 15 LMICs participating in GATS between 2008 and 2011. This study involved secondary analysis of GATS data from 15 LMICs. GATS is a nationally representative cross-sectional household survey of non-institutionalized adults aged 15 years and over (World Health Organization, 2013c). It is considered to be the global standard for monitoring adult tobacco use and key tobacco control indicators. GATS employs standardized survey methodology with a few country-specific variations in the questionnaire, and is designed to collect household as well as individual level data. Multi-stage cluster sampling design is employed in GATS to select a nationally representative study sample. Between 2008 and 2011, the first round of GATS was implemented in 17 LMICs in five WHO regions (Centers for Disease Control and Prevention, 2013a). Country-specific, anonymous GATS data for 15 of the 17 LMICs (all but Indonesia and Malaysia) was freely available from the CDC GTSS Data website, which was used for secondary data analysis.

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The literature suggests that health professionals need


The literature suggests that health professionals need

to undertake cross-cultural communication training to improve their interpersonal skills for interacting with Indigenous people, to encourage greater respect towards Indigenous culture and to help understand the dissonant world views of health and illness between Indigenous people and mainstream society.8, 12 and 16 Whilst this type of training may be useful to some extent, it is unlikely to result in entirely competent health practitioners who appreciate the diversity of Indigenous people and their culture, and who are able to interact with all Indigenous people in an appropriate and respectful manner. The heterogeneity of Indigenous Australians means there is not one set-recipe for communicating

with Indigenous people10 and cross-cultural practice requires more than just an understanding and awareness of different cultures NSC 683864 in vitro and health perspectives. The authors’ therefore argue for a more nuanced approach – one that places greater click here focus on the reflexive skills of the practitioner and that encourages health professionals to consider each individual’s world view of health and illness and the factors that conceptualise people’s health experiences.10 The Australian Physiotherapy Council states the need for critical self-reflection by physiotherapists to acknowledge their own cultural beliefs and values,

and any assumptions that they bring to the clinical interaction.11 The physiotherapy profession has constructed its own identity, incorporating values and interpretations of what are believed to be good practice.19 However, it is important to reflect on these values and acknowledge personal biases and ethnocentricity Rutecarpine – the unconscious belief that these interpretations and assumptions are correct – and how this may impact on clinical interaction.19 This includes recognising the influence of the dominant culture and how conscious and sub-conscious use of power may impact on relationships with clients and on clinical decisions.20 Critical self-reflection is paramount to avoid essentialising Indigenous culture and to ensure that physiotherapists communicate and interact with Indigenous people appropriately and effectively. As with other population groups, there is growing recognition of the importance of adopting a person-centred approach in Indigenous healthcare and to acquire a broader understanding of the Indigenous health experience from the person’s perspective.21 The person-centred approach, which is supported by the Australian Physiotherapy Council,11 was advocated by Enid Balint over 40 years ago to better understand the whole person, including their social world and individual needs, rather than merely fitting them into predetermined criteria based on illness.

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MRI demonstrated a lack of recurrent tumor up to 1 year following

MRI demonstrated a lack of recurrent tumor up to 1 year following surgery (Fig. 1A). Neurological side effects of this therapy were moderate and resolved within 3 months. The treated dog experienced transient focal neurologic signs that became more severe with each subsequent vaccine. Specifically, focal seizures, left hemiparesis, and acute blindness as assessed by lack of menace

response in the left eye were documented after the fourth and fifth vaccinations. Left hemiparesis and left-sided blindness became apparent 3 days after the forth and fifth vaccination, and resolved within a week in each instance. In addition, during therapy, the dog exhibited circulating lymphopenia, peripheral lymphadenopathy around the vaccination site, and elevation of gamma glutamyltransferase

(GGT) and alanine aminotransferase (ALT) that were not Venetoclax nmr present prior to treatment (Table 1). Although selleck chemicals llc the elevation of such liver enzymes may have been induced by anti-seizure medication (see Section 4), the timing of the other neurological symptoms 3 days after the fourth and fifth vaccination indicate a possible relationship with the vaccines. In order to detect IgG responses specific to the dog’s own tumor, tumor cell lysates from the autologous cells grown in culture were subjected to SDS-PAGE. Autologous serum harvested before or after vaccinations was used as a primary antibody and specific IgG was detected using anti-canine IgG antibody. Western blot analysis revealed that this dog did not have an appreciable pre-existing tumor-reactive IgG response, and there was no signal from normal canine serum used as a control. By 2 weeks after the first vaccination, IgG reactive to two proteins approximately 50–65 kDa in weight was detected (Fig. 2A). This response remained unchanged 2 weeks later at day 65, but increased in breadth of antigens recognized as subsequent vaccinations were administered. A memory IgG response was induced to three separate tumor antigens,

as revealed by three bands on Western Blot using serum harvested over 100 days following the last vaccination others (Fig. 2A). Upon recognition of their cognate antigen, CTLs elaborate IFNγ and release cytotoxic granules; the proteins CD107a, CD107b, and CD63 within the granule mobilize to the cell surface during degranulation (reviewed in [26]). Accordingly, CD107 cell surface mobilization measured by flow cytometry demonstrates a linear correlation with tumor cell lysis [27], and CD107 has been used to monitor CTL responses in melanoma patients treated by vaccination [28]. Because the number of peripheral blood mononuclear cells (PBMCs) obtained from this dog was limited, we elected to employ this flow cytometry-based assay to detect CTL degranulation and IFNγ production. PBMCs frozen at various time points before and after surgery were thawed and analyzed identically and simultaneously to eliminate interexperiment variability.

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The SPADI has since been used in both primary care on mixed diagn

The SPADI has since been used in both primary care on mixed diagnosis (Beaton et al 1996, MacDermaid et al 2006) and surgical patient populations including rotator cuff disease (Ekeberg et al 2008), osteoarthritis, and rheumatoid arthritis (Christie et al 2010), adhesive capsulitis (Staples et al 2010, Tveita et al 2008), joint replacement surgery (Angst et al 2007), and in a large population-based study of shoulder symptoms (Hill et al 2011). The SPADI is available free of charge at several sites, eg, www.workcover.com/public/download.aspx?id=799. Instructions to the client and scoring: In the original version the patient was instructed SB431542 to place a mark on the VAS for each item

that best represented their experience of their shoulder problem over the last week (Roach et al 1991). Each subscale is summed and transformed to a score out of 100. A mean is taken of the two subscales to give a total score out of 100, higher score indicating greater impairment or disability. In the NRS version (Williams et al 1995) the VAS is replaced by a 0–10 scale and the patient is asked to circle the number that best describes the pain or disability. The total score is derived in exactly the same manner as the VAS version. In each subscale patients may mark one item only as not applicable Selleck GDC-0199 and the item is omitted from the total score. If a patient

marks more than two items as non applicable, no score is calculated (Roach et al 1991). Reliability and validity: Reproducibility of the SPADI in the original description was poor, with an intraclass correlation coefficient (ICC) of 0.66. A more recent systematic review has found reliability coefficients of ICC ≥ 0.89 in a variety of patient populations (Roy et al 2009). Internal

consistency is high with Cronbach α typically exceeding 0.90 (Roy et al 2009, Hill et al 2011). The SPADI demonstrates good construct validity, correlating well with other region-specific shoulder questionnaires (Paul et al 2004, Bot et al 2004, Roy et al 2009). It has been GBA3 shown to be responsive to change over time, in a variety of patient populations and is able to discriminate adequately between patients with improving and deteriorating conditions (Beaton et al 1996, Williams et al 1995, Roy et al 2009). No large floor or ceiling effects for the SPADI have been observed (Bot et al 2004, Roy et al 2009). The minimal clinically important difference has been reported to be 8 points; this represents the smallest detectable change that is important to the patient (Paul et al 2004). When the SPADI is used more than once on the same subject, eg, at initial consultation and then at discharge, the minimal detectible change (MDC 95%) is 18 points (Angst et al 2008, Schmitt et al 2004). Thus some caution is advised with regard to repeated use of the instrument on the same patient.

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Cerebral ischaemia is a powerful inducer of the UPR [37], and sub

Cerebral ischaemia is a powerful inducer of the UPR [37], and subjecting JEG-3 cells to hypoxia-reoxygenation causes phosphorylation of eIF2α

[25]. This situation may be made worse by changes in posture, which in the bipedal human can influence uterine blood flow [38], or heightened uterine contractility, as maternal placental blood is reduced during a contraction [39]. The intervening steps in vivo are unclear at present, but various possibilities exist. Episodes of ischaemia will deplete intracellular concentrations of glucose, which may restrict normal glycosylation within the ER, activating the UPR. Alternatively, ischaemia will reduce intracellular levels of ATP, compromising the functioning of the GRP chaperone proteins, buy Trametinib and possibly also the Ca2+-ATPase ionic pumps within the ER membrane. Ischaemia may also have a more direct effect on calcium release from the ER by altering the redox balance within the cell, affecting thiol groups on the calcium channel proteins [40]. Calcium imbalance may further result from competitive binding of GRP78 to misfolded proteins, for under normal conditions GRP78 serves to plug unoccupied translocons, preventing leakage. Loss

of calcium from the ER lumen will compound the situation by compromising the protein folding machinery, and by activating calcium dependent signalling pathways within the cytsol. Ultimately, these could lead to opening of the mitochondrial membrane transition pore, with subsequent loss of mitochondrial function and generation of ROS. We have previously demonstrated that hypoxia-reoxygenation of villous Lumacaftor mw explants leads to opening of the pore, and activation of apoptosis within the syncytiotrophoblast [41]. Further work is required to tease apart these various possibilities, but the complex interactions between oxidative and ER stress mean that once one is initiated the other is likely to follow soon after through

feed-forward mechanisms. In many isothipendyl instances pathological activation of the UPR is a one-off event, following for example stroke or myocardial infarction. As mentioned earlier, phosphorylation of eIF2α and inhibition of protein synthesis are usually transient events, for activation of ATF4 leads to upregulation of the phosphatase GADD34. However, the precipitating vascular insult to the placenta in pre-eclampsia is likely to be of a lower grade than that in stroke, and also of a repetitive nature. To mimic this in vitro we have exposed JEG-3 cells to repetitive cycles of hypoxia-reoxygenation and observed sustained phosphorylation of eIF2α and activation of the UPR. We predict therefore that the ER stress is of a chronic nature, dating most likely from the time of onset of the maternal circulation at the end of the first trimester. The consequences for placental function are manifold, and are just beginning to be explored [42].

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This is accomplished by increasing the concentration of acetylcho

This is accomplished by increasing the concentration of acetylcholine through reversible inhibition of its hydrolysis by acetylcholinesterase. The recommended

initial dose of donepezil is 5 mg taken once daily. Donepezil is well absorbed with a relative oral bioavailability of 100% and reaches peak plasma concentrations (Cmax) approximately 3–4 h S3I-201 purchase after dose administration. In humans, donepezil is metabolized mainly by the hepatic cytochrome P-450 2D6 and 3A4 isozymes. 2 Elimination of donepezil from the blood is characterized by a dose independent elimination half-life of about 70 h. 3 and 4 Because plasma donepezil concentrations are related linearly to acetylcholinesterase inhibition, 5 plasma donepezil concentration is a useful tool to predict donepezil efficacy. In the literature, methods have been reported for the quantification of donepezil in biological fluids. Methods are reported for the quantification of donepezil from biological

matrix using high-performance liquid chromatography (HPLC) equipped with an ultraviolet detector,2 and 3 fluorescence detector4 and mass spectrometric1, 6 and 7 detector. Methods are also reported for the quantification of enantiomers of donepezil from human plasma.8, 9 and 10 Other methods are reported with estimation of donepezil in plasma by capillary electrophoresis,11 hydrophilic interaction chromatography-tandem mass spectrometry,12 direct measurement,13 automated Florfenicol extraction.14 The HPLC methods used to determine donepezil in human plasma are insensitive because

of the lower limit of quantification (LOQ of >1.0 ng/ml). Some of the MAPK Inhibitor Library reported methods1, 4, 6, 10, 13 and 14 utilized analogue internal standards like diphenhydramine, lidocaine, pindolol, loratadine, escitalopram, etc. and are validated with different calibration curve ranges for the estimation of donepezil from rat plasma, human plasma and other biological fluids. Usage of labelled internal standards is recommended during the estimation of compounds from the biological matrices to minimize the matrix effects associated with the mass spectrometric detection. Bioequivalence and/or pharmacokinetic studies become an integral part of generic drug applications and a simple, sensitive, reproducible validated bioanalytical method should be used for the quantification of intended analyte. Bioequivalence studies for the donepezil needs to be performed with the dosage of 10 mg and 23 mg tablets to support the generic abbreviated new drug applications. For the pharmacokinetic and bioequivalence studies, quantification of donepezil was sufficient and quantification of its metabolites shall not be required. During the bioequivalence studies, appropriate lower limit of quantification needs to be used to appropriately characterize the concentration profile including the elimination phase.

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Western blots were probed using murine sera raised to recombinant

Western blots were probed using murine sera raised to recombinant proteins based on the individual MSP1 block 2 types [11] and [15]. Bound antibody was detected with horseradish peroxidase-conjugated rabbit anti-mouse secondary antibody

(DAKO), and bands visualized using 5 ml per blot of stabilized TMB (3,3′,5,5′-tetramethylbenzidine) substrate (Promega, UK). Groups of five CD-1 outbred mice were immunized (Northwick Park Institute for Medical Research, UK) with each antigen formulated in the ImjectAlum adjuvant (Perbio Science, Cheshire, UK). Each polyvalent hybrid protein was diluted with Selleck Luminespib phosphate-buffered saline (PBS) to a concentration of 1 mg ml−1, and 3 volumes of Imject Alum Z-VAD-FMK added and allowed to mix for 30 min at room temperature. Each antigen–adjuvant mixture was administered intra-peritoneally, each mouse receiving 50 μg protein per dose in a final volume of 200 μl. Three doses were administered

at monthly intervals, and blood was collected before immunization and 2 weeks after each dose (on days 14, 42, and 70). The purified polyvalent hybrid antigen (+)T-K1SR-R033-Wellcome (antigen 6, Fig. 1A) was used to immunize New Zealand white rabbits (Pettingill Technology Limited, UK). Five rabbits received 200 μg of purified protein intramuscularly enough at days 0, 14, 28, 42, 56 and 70 following a 77 day protocol with one rabbit receiving adjuvant with PBS only as a control (Freund’s complete adjuvant was used on day 0 immunization, Freund’s incomplete adjuvant for boosting immunizations). Test bleeds were taken on days 35, 49 and 63, final bleeds were collected on day 77. Ten P. falciparum isolates were

cultured, including 6 with K1-like MSP1 block 2 sequences (3D7, T9/96, T9/102, D6, K1, Palo Alto), 3 with MAD20-like block 2 sequences (Wellcome, MAD20, Dd2), and R033 representing the R033-like block 2 type that has minimal subtypic polymorphism. Each was identified and discriminated by sequencing of MSP1 block 2. Parasite cultures were grown under standard conditions to a parasitemia of 4–10% (predominantly schizont stage although asynchronous) and cells washed twice after centrifugation before resuspension in PBS/1% BSA, for preparation of IFA slides. Specific antibody reactivities to each of the parasite isolates were assessed following previously described methods [22]. Parasites were air-dried onto multiwell IFA slides (Hendley, Essex, UK), fixed with 4% formaldehyde and tested with serial doubling dilutions of murine sera (1/50 to 1/409,600) in PBS with 1% bovine serum albumin (15 μl/well) and incubated for 30 min. Biotinylated anti-mouse or anti-rabbit IgG (Vector Laboratories Inc.

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The study protocol was approved by the ethics committee of the He

The study protocol was approved by the ethics committee of the Helsinki University Central Hospital and the Finnish Medicines Agency. The study protocol was registered in the International Standard Randomised Controlled Trial Number Register (ISRCTN68125331). Written informed consent was obtained from all study subjects. The patients enrolled in this study were treated in the Division of Infectious Diseases, Helsinki University Central Hospital. Thirty healthy

Finnish born volunteers (18 females, 12 males, aged 18–62 years, mean age 32 years), four patients with typhoid fever (two females, two males, aged 22–29 years) and one with paratyphoid fever (female, 30 years) were enrolled. Of the patients with typhoid fever, two were Finnish born travelers to India and South-America, one was an applicant Kinase Inhibitor Library cell assay BGB324 order for asylum from Sri Lanka and one was an immigrant from Nepal who had visited relatives in his home country. The last patient was having an infection relapse one month after the first episode. The patient with paratyphoid A fever was an immigrant from India who had visited relatives in her home country. Typhoid and paratyphoid fever were diagnosed on the basis of blood cultures. None of the vaccinees had a previous history of receiving typhoid

vaccine or having enteric fever. They were given the oral Salmonella Typhi Ty21a vaccine containing ≥2 × 109 live bacteria/capsule (Vivotif®, Crucell, Leiden, The Netherlands, lot 3001777) administered one capsule per day on days 0, 2 and 4, as recommended by the manufacturer. Peripheral venous blood was drawn on days Amisulpride 0 and 7 after vaccination or 7–10 days after the onset of symptoms of the infection. To include as many antigenic structures as possible, whole bacteria of strains Salmonella Typhi (Vsa61), Salmonella

Paratyphi A (RHS6716), B (RHS6744), C (ATCC-13428) and Salmonella Egusi (RHS6854) were used as antigens in the ELISPOT assay. Salmonella Paratyphi C strain was from the American Type Culture Collection (ATCC, Manassas, VA, USA), while the other strains were from the National Institute for Health and Welfare, Helsinki, Finland. Bacteria were cultured on nutrient agar plates to determine their concentration in the suspension, and formalin-killed as described previously [20]. For ELISPOT assays, the concentration was adjusted to 109 bacteria/ml in PBS (phosphate buffered saline). PBMC were separated using Ficoll-plaque density gradient centrifugation as described previously [20]. The analyses of HR expressions were carried out for 15 vaccinees and for the four patients with enteric fever as a primary infection. Only one strain per person could be analyzed because of limited numbers of PBMC.

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