This truncated TSOL16A cDNA (herein referred to as TSOL16 with re

This truncated TSOL16A cDNA (herein referred to as TSOL16 with respect to the cDNA and encoded protein) was cloned directionally into the EcoRI and XhoI sites of pGEX-1TEX and transformed into E. coli JM109 strain by electroporation. Use of the pGEX plasmid allowed

expression and purification of TSOL16 as a fusion with glutathione S-transferase (GST) [15]. The truncated TSOL16 cDNA was excised from pGEX-1 by digestion with EcoRI and XhoI, HKI-272 order and cloned into EcoRI/SalI-digested pMAL-C2. The pMAL-C2 plasmid allowed expression and purification of TSOL16 as a fusion with maltose binding protein (MBP) [16]. The plasmid construct was transformed into E. coli JM109. The TSOL45-1A protein was cloned into the pGEX and pMAL-C2 plasmids, and expressed in E. coli as a fusion protein with GST and MBP as described in [4]. The TSOL45-1A fusion proteins lacked 16 N-terminal amino acids that encoded a predicted secretory signal. The TSOL45-1B

cDNA was originally cloned from T. solium oncosphere mRNA as described in [7]. TSOL45-1B lacked exon II of the TSOL45-1 gene. PCR amplification was used to produce a cDNA construct that encoded a protein also lacking the 16 N-terminal amino acids of the secretory signal. The following PCR primers were used to amplify TSOL45-1B for cloning into pGEX and pMAL as described above: 5′CCG GAA TTC GGA AAC CAC AAG GCA ACA TC3′; 5′CCG CTC GAG GGA AAT GGG CAT TGA CCG3′. E. coli SAR405838 molecular weight cultures expressing TSOL16, TSOL45-1A and TSOL45-1B were prepared and recombinant fusion proteins were purified as detailed in [14]. Freeze-dried aliquots of antigens were prepared by the addition of Quil A adjuvant (1 mg per dose) and a much sixfold (w/w) amount of maltose as a stabilizing agent for transport to Lima, Peru, where

the vaccine trial was conducted. Aliquots of GST and MBP, for use as negative controls, were also prepared for the vaccine trial. The antigens were reconstituted in sterile de-ionized water immediately prior to vaccination of pigs. The purified GST and MBP fusions of TSOL16, TSOL45-1A and TSOL45-1B were tested in a pig vaccine trial against challenge infection with T. solium. The study was reviewed and approved by the Animal Ethics Committee of the School of Veterinary Medicine, Universidad de San Marcos, Lima, Peru. Twenty 8-week old piglets were obtained from a cysticercosis free farm located in Huaral, Lima. Animals were divided into four groups of 5 pigs each. All animals were vaccinated against Classical Swine Fever prior to the start of the trial. Each pig received 200 μg of antigen and 1 mg Quil A (Brenntag Biosector, Denmark) per immunization in a 1 ml dose. Immunizations were given intramuscularly in the right hind-quarter via a 0.9 mm × 38 mm needle and 1 ml syringe (Becton Dickinson, U.K.). Piglets received their first immunization with recombinant antigen prepared as a GST fusion.

1A) (P < 0 0001), and greater with the 97 day interval than the 5

1A) (P < 0.0001), and greater with the 97 day interval than the 57 day interval (P = 0.0006). The antibody response induced by protein–protein (P–P) vaccination was markedly variable with three mice mounting high responses comparable to those receiving A–P immunization, and three very weakly responding mice ( Fig. 1A and B). There was no significant difference Crenolanib between median antibody responses following protein–protein, adenovirus–MVA and adenovirus–protein regimes after a 57 day dose interval (P = 0.37 by Kruskal–Wallis test), but there was a clear increase in the variance of the

response after two shot protein regimes compared to viral-vector containing regimes. In contrast with the antibody results, greater

percentages of IFNγ+ CD8+ T cells were detected by ICS 14 days after A–M immunization than A–P, and the 57 day dose interval was superior (P < 0.0001 for both comparisons) ( Fig. 1A and B). Clear boosting of CD8+ T cell responses by MVA was evident at both dose intervals. As expected, given the lack of the CD8+ T cell epitope in the MSP119 protein sequence in BALB/c mice [5], CD8+ T cell responses were not detectable following P–P vaccination. Additional experiments in C57BL/6 mice (in which a CD8+ T cell epitope is present in the MSP119 protein [5]) confirmed that, in contrast to the A–M regime, P–P Talazoparib vaccination did not induce a CD8+ T cell response detectable by IFNγ splenic ELISPOT or peripheral blood ICS, and that CD8+ T cell responses were unaltered by A–P immunization as compared to adenovirus priming alone ( Fig. 1C and D). CD8+ T cell responses after A–P immunization of either mouse strain thus presumably represent the contracting or effector memory CD8+ T cell response induced Tryptophan synthase by the adenovirus. We subsequently compared the immunogenicity of three-component sequential adenovirus–MVA–protein (A–M–P) and adenovirus–protein–MVA (A–P–M) regimes to two-component regimes (Fig. 2 and Fig. 3). The kinetics of the responses induced by these regimes were markedly different. We found that addition of

protein to adenovirus–MVA (A–M–P) was able to boost antibody but not CD8+ T cell responses (again as would be predicted due to lack of the T cell epitope in this protein) (Fig. 2A), while addition of MVA to adenovirus–protein (A–P–M) boosted CD8+ T cell responses but not antibody titer (Fig. 2B). Total IgG responses to A–M–P and A–P–M were significantly higher than those to A–M (P < 0.05 by ANOVA with Bonferroni post-test), with no significant differences between the responses to A–M–P, A–P–M and A–P (P > 0.05, Fig. 3A). There were no statistically significant differences in CD8+ T cell responses between A–M–P, A–P–M and A–M regimes (P > 0.05 by ANOVA with Bonferroni post-test, Fig. 3B). In general, any two- or three-component regime including AdCh63 and MVA induced maximal CD8+ T cell responses as measured in the blood.

This burden is also similar to earlier studies on rotavirus burde

This burden is also similar to earlier studies on rotavirus burden in hospitalized AGE cases [5] and [6]. We found G1 and G2 as the most common G types, P[4] and P[8] as the most common P types and G1P[8] and G2P[4] as common GP types. Some rotavirus samples could not be typed for selleck G and/or P type. The most common G/P/GP types found in this study are similar to other Indian studies (including IRSN) conducted in children hospitalized with RVGE [2], [3], [4],

[5] and [6]. Our results show that G12 comprised 6.4% of rotavirus strains: a finding in concordance with IRSN [4] and [6]. G12 strain was first detected in India in 2001 and over the decade has been increasingly reported in recent Indian studies [4], [6], [17] and [18]. More than 75% of the children enrolled in the study were in the age group of less than 2 years. This reflects the age profile of diarrhea burden in India, where majority of the diarrhea episodes in children under 5 years of age are reported to occur in children of age less than 3 years [19] and [20]. In our study, mean age of RV positive

subjects was lower compared to RV negative subjects and majority of RVGE (85%) cases occurred in children ≤24 months of age. The difference between rotavirus and non-rotavirus groups was significant w.r.t. age distribution – result similar to previous observations of the epidemiologic profile of rotavirus infection in India [4] and [5]. In IRSN, it was observed that the mean age of RV positive children was significantly lower than RV negative children. In addition to younger Cabozantinib mouse age of RVGE subjects, our results also indicate that RV positive subjects experience severe and multiple AGE symptoms. We found that more than half of the RVGE cases were severe by Vesikari scale (77.2%) while a few were severe by Clark scale (3.9%). Similar distribution was seen in non-RVGE cases. Higher proportion of severe cases in our study may be due to late referral of the subjects to OPDs after disease

onset. A 10 district survey in India by UNICEF titled “Management Practices of Childhood Diarrhea in India” has reported that in India in rural as well as urban areas, there is delay of at least 1 day between onset of diarrhea and time of seeking medical care outside home. The report also mentions that parents Dipeptidyl peptidase took the child outside home for managing diarrhea when child had too many stools, appeared very weak, did not eat anything, and diarrhea continued for too long [20]. It is likely therefore that majority of parents take their child to health care setting when diarrhea becomes severe. We used Clark and Vesikari scale for categorizing acute gastroenteritis into different severity levels. This categorization is dependent on multiple factors like study methodology such as where, how and when data is collected, active or passive method surveillance and frequency, timing, method of assessment in active studies.

The measurement of the extracellular L-Glu concentration in the m

The measurement of the extracellular L-Glu concentration in the medium was performed according to the methods previously

described (8). Real-Time Quantitative RT-PCR, Western Pictilisib chemical structure blotting, immunocytochemistry were also performed according to the methods previously described (8). The microglia culture was treated with LPS for 24 h in the presence or absence of antidepressants and the concentration of L-Glu in the medium was measured. All sets of the experiments were repeated in triplicate. All procedures described above were in accordance with institutional guidelines. In the previous report, we showed that the expression level of astrocytic L-Glu transporters was decreased Lumacaftor cell line in the astrocyte-microglia-neuron mixed culture in LPS (10 ng/ml, 72 h)-induced inflammation model without cell death (8). We first compared the effects of various groups of antidepressants, i.e., selective serotonin reuptake inhibitors (SSRIs) (paroxetine, fluvoxamine, and sertraline), serotonin–norepinephrine

reuptake inhibitor (SNRI) (milnacipran), and tricyclic antidepressant (TCA) (amitriptyline), on the decrease in the astrocytic L-Glu transporter function in this inflammation model. To quantify L-Glu transport activity, we measured the concentration of L-Glu remaining 30 min after changing the medium to the one containing 100 μM of L-Glu. In each set of experiment, LPS-induced decrease in the L-Glu transport activity was stably reproduced (Fig. 1A–E). Among antidepressants, only paroxetine prevented the LPS-induced decrease in L-Glu transport activity (Fig. 1A). The effect was concentration-dependent and reached significant at 1 μM. The other antidepressants had no effects (Fig. 1B–E). Typical image of the astrocyte-microglia-neuron mixed culture was shown in Fig. 1F. We have clarified that LPS-induced medroxyprogesterone decrease in L-Glu transport activity was caused by the decrease in the expression level of GLAST, a predominant L-Glu transporter in the mixed culture, in both of mRNA and protein levels (8). In this study, LPS-induced decreases in the

expression of GLAST, were reproduced at both of mRNA (28.8 ± 4.7% of the control) and protein (69.5 ± 4.7% of the control) levels (Fig. 1G, H). We then examined the effects of paroxetine on the LPS-induced decrease in the L-Glu transporter expression. Paroxetine significantly prevented the decreases at both of mRNA (28.8 ± 4.7 to 49.6 ± 3.3%; n = 10) and protein (from 69.5 ± 4.7% to 91.0 ± 5.1%; n = 5) levels ( Fig. 1G, H). As is shown in Fig. 1, fluvoxamine and sertraline, the other SSRIs in this study, did not affect the decrease in L-Glu transport activity, suggesting that paroxetine revealed the effects through the mechanisms independent of its inhibitory effect on serotonin selective transporter.

S ) (Ogden et al , 2012) Public health authorities are beginning

S.) (Ogden et al., 2012). Public health authorities are beginning to look for cost-effective ways to reduce this epidemic. Increased physical activity is a candidate strategy because of its numerous health benefits, including the potential to attenuate cardiovascular disease and diabetes risk ( Kahn et al., 2002, Norman et al., 2006 and Task Force on Community Preventive Services (USTFCPS), 2001).

Research has shown that there is a positive association between proximity to parks/recreational facilities and increased physical activity levels ( Roemmich et al., 2006 and Sallis et al., 2011). Programming INK 128 datasheet and group activities, for example, have been found to be related to increased usage of school facilities and improved levels of moderate-to-vigorous physical activity ( Lafleur et al., 2013). Having convenient, reliable access to Apoptosis Compound Library ic50 open space/recreational areas or programing that encourages physical activity, however, can be challenging, especially for under-resourced communities ( Marie, 2007, Powell et al., 2006 and Spengler et al., 2007). Shared-use agreements (SUAs) where school property (i.e., the grounds, facilities, or both) and programming are shared between schools and

community-based entities represent a strategy to address this public health problem. A shared-use agreement outlines an agreement between two or more parties that details and enumerates each party’s responsibilities in the partnership. Shared-use encompasses a diverse array of agreement types, including joint-use agreements (JUA) and Memoranda of Understanding (MOUs). These contractual documents may be legally binding or non-binding; but whether or not they are legally binding does not diminish their potential benefits. A formal agreement adds value to each partnership by laying out the expectations of the entering parties, reducing the odds that the relationship would dissolve prematurely. School grounds offer clean, protected, and often underutilized space that community members can use for physical activity

(Maddock et al., 2008). Communities that seek to promote physical activity and improve access to recreational space can partner with school districts. Non-profit organizations are also important CYTH4 partners as they often receive outside funding to provide programming (Lafleur et al., 2013). SUAs offer the opportunity for both parties to clarify their intent and roles in the partnership, as well as to identify their individual interests. Even when state laws generally provide schools strong protection against liability for injuries to recreational users of school properties (California Tort Claims Act, 2012), the perceived threat of tort liability remains an important deterrent to schools’ decisions to participate (Spengler et al., 2007 and Zimmerman et al., 2013).

The barrier properties of the skin membrane depend on the molecul

The barrier properties of the skin membrane depend on the molecular organization of the SC components. Considering this, we employed SAXD and WAXD to investigate the effect of glycerol and urea on both the organization of the SC extracellular lipid lamellae and on the soft keratin

structures. The results from the SAXD and WAXD measurements at 32 °C are presented in Fig. 2A and B, respectively. We start by concluding that the results obtained for the SC sample without glycerol or urea are in good agreement with previous SAXD and WAXD studies on hydrated pig SC (Bouwstra et al., 1995). Further, it is shown that the JAK activation SC pretreated in glycerol or urea formulations give rise to similar diffraction curves as the SC pretreated in neat PBS solution. All SAXD curves in Fig. 2A have one broad peak centered around Q = 1.0 nm−1 (6.3 nm in d-spacing). The strong diffraction at low Q is attributed to protein structures of the SC ( Bouwstra et al., 1995 and Garson et al., 1991), which obscures the diffraction pattern of any lipid structures in this region. However, centered around Q = 0.5 nm−1 (12.6 nm in d-spacing) a shoulder is present in the descending diffraction curves, which implies that the peak around 6.3 nm in d-spacing is a learn more 2nd order peak of

a lamellar phase with approx. 12.6 nm in d-spacing. When the SC sample has been pretreated in the formulation that contain urea (bottom curve), the shoulder around Q = 0.5 nm−1 is nearly absent, and the intensity of the peak around Q = 1.0 nm−1 is weaker compared to the other samples. A weak shoulder centered around Q = 1.4 nm−1 (4.5 nm in d-spacing) is present in all diffraction curves in Fig. 2A. In the literature, the same peak at 4.5 nm has been interpreted as the 2nd order of a 9 nm periodicity lamellar phase ( Bouwstra et al., 1995). However, no signs of a 1st

order peak of this 9 nm lamellar phase was observed here. Considering that all reflections are diffuse and broad it cannot be ruled out that all of the above peaks/shoulders belong to the same lamellar found phase with repeat distance of approx. 12.6 nm. Finally, a peak centered around roughly Q = 1.8 nm−1 (3.4 nm in d-spacing) is observed in all diffraction curves, which is attributed to phase separated crystalline cholesterol ( Bouwstra et al., 1995). Fig. 2B shows WAXD data for the corresponding conditions as in Fig. 2A. A distinct peak at approx. Q = 15.2 nm−1 (0.41 nm in d-spacing) is present in all diffraction curves, irrespective of pretreatment formulation. This peak corresponds to hexagonal packed lipid carbon chains. No signs of orthorhombic packing was observed under any conditions (i.e., no peak was present at approx. Q = 17 nm−1 or 0.37 nm in d-spacing), which is in agreement with previous studies on pig SC ( Bouwstra et al., 1995 and Caussin et al., 2008).

Our findings suggest that clinicians may not always find retinal

Our findings suggest that clinicians may not always find retinal hemorrhages in abused children. Moreover, our study perhaps underestimated the incidence

of such findings since we focused on injuries found to be severe enough to cause death. The survivors may have had subdural hemorrhages detectable by magnetic resonance imaging (MRI). The MRI can be a vital tool, with great sensitivity and specificity, for identifying those infants who have brain subdural hemorrhage but lack retinal hemorrhages and who would otherwise be overlooked for abusive MK-8776 mouse head trauma.23 Retinal hemorrhages in our study were also found to be proportionately more frequent in children younger than 16 months of age compared to infants older than 16 months. Our study is similar to one in which children younger than 1 year were found more likely to have retinal hemorrhages.24 This same study also demonstrated a “dome-shaped hemorrhagic lesion” in the macula “that elevates the internal limiting membrane,” essentially describing the perimacular ridge. This is similar in appearance to cherry hemorrhages typically

located peripherally. To Ivacaftor chemical structure our knowledge, the cherry hemorrhage has not been previously described. Found in 40% of our abusive head trauma eyes and demonstrated using gross, histopathologic, and TEM examinations (Figure 4), the cherry hemorrhage is a distinct hemorrhagic lesion often confined to the equatorial retina that can be seen by indirect ophthalmoscopy. Microscopically, it is similar to the perimacular

ridge with a dome of torn ILM over a large hemorrhage. Furthermore, this lesion was found only in eyes that had a torn ILM and concurrent retinal hemorrhages extending to the ora serrata. The threshold of acceleration–deceleration forces necessary to produce bleeding throughout the retina (ora-extended) is likely lower than that for creating the cherry hemorrhage. Neither a cherry hemorrhage nor an ora-extended hemorrhage was found in control eyes. Thus, the cherry hemorrhage is one more robust criterion for identifying tuclazepam abusive head trauma. Our findings most strongly corroborate the role of vitreoretinal traction. Other, less-substantiated hypotheses include increased intrathoracic pressure, increased intracranial pressure, and retinal hypoxia.22 Indeed, animal models have determined a limited role for retinal hypoxia in the presence of retinal hemorrhages.25 This finding parallels the absence of retinal hemorrhages found clinically in hypoxic children.22 Laterality of findings is an important consideration when faced with a diagnosis of abusive head trauma. All eyes in our series were proportionately more likely to have bilateral than unilateral pathology. However, at least 1 unilateral presentation for each finding, except subdural hemorrhage, was found in all cases.

Participants were enrolled sequentially in three steps preceded b

Participants were enrolled sequentially in three steps preceded by a safety review (Fig. 1). They were randomized find more (1:2:2:2:2:2:2, block size 4 [step 1], 7 [step 2] and 5 [step 3]) using a central internet randomization system (SBIR) to receive a two-dose primary vaccination series with one of six investigational vaccine formulations (GlaxoSmithKline Vaccines) or a single dose of the 23-valent pneumococcal polysaccharide vaccine (23PPV; Pneumovax23™, Sanofi Pasteur

MSD) followed by placebo (150 mM NaCl) ( Fig. 1; supplementary methods). All vaccines and the placebo were administered intramuscularly into the deltoid region of the non-dominant arm. Two investigational vaccines contained 10 or 30 μg of dPly alone (dPly-10 and dPly-30, respectively). Two other formulations contained Lumacaftor in vitro both dPly and PhtD, each at a dose of 10 μg (dPly/PhtD-10) or 30 μg (dPly/PhtD-30). The remaining two formulations contained the 10 PHiD-CV PS-conjugates (serotypes 1, 4, 5, 6B, 7F, 9V, 14, 18C, 19F and 23F) [18], in combination with 10 or 30 μg of both dPly and PhtD (PHiD-CV/dPly/PhtD-10 and PHiD-CV/dPly/PhtD-30).

Production of PhtD and dPly is described in supplementary methods. The control group received one dose of 23PPV, containing 25 μg of each capsular polysaccharide for pneumococcal serotypes 1, 2, 3, 4, 5, 6B, 7F, 8, 9N, 9V, 10A, 11A, 12F, 14, 15B, 17F, 18C, 19A, 19F, 20, 22F, 23F and 33F, and placebo (150 mM NaCl) as a second dose. Participants from the dPly/PhtD-10 and dPly/PhtD-30 groups were invited to participate in the booster vaccination study, to receive a booster dose 5–9 months after completion of the two-dose primary series. Solicited local and general symptoms were recorded during the 7-day post-vaccination period and unsolicited adverse events (AEs) during the 31-day post-vaccination period. Symptom intensity was graded on a scale of 1 (mild) to 3 (severe). Grade 3 symptoms were defined as follows: for redness or swelling, a diameter >50 mm; for fever, oral temperature >39.5 °C; and for all

other events, preventing normal activity. Serious adverse events (SAEs) were recorded throughout the duration of each study, and were defined as any medical occurrence that resulted in death, disability or incapacity, was life-threatening, required Electron transport chain hospitalization, or any congenital anomaly or birth defect in the descendants of a study participant. Blood samples for immunogenicity assays were collected before primary and booster vaccination, and 1 month after each dose. Serum samples were stored at −20 °C until analysis at GlaxoSmithKline’s laboratory, Rixensart, Belgium and SGS laboratory, Wavre, Belgium. Antibodies were quantified using an in-house multiplex assay coated with protein D, Ply (non-detoxified) and PhtD (supplementary methods), with assay cut-offs of 112 LU/mL for anti-PD, 599 LU/mL for anti-Ply and 391 LU/mL for anti-PhtD.

5% biochar-amended soil presented unobvious changes throughout th

5% biochar-amended soil presented unobvious changes throughout the duration, and a gradual decrease in porosity appeared in the 5% biochar-amended soil. Fig. 2g indicates that MWD of soil aggregation find more was consistently higher for the biochar-amended soils than the control after incubation of 21 d; however, significant differences between the amended soils and the control were found after incubation of 84 d. An obvious peak that occurred at 21 d was found

for all treated soils. Furthermore, applying biochar to the soil caused a significant increase in the saturated hydraulic conductivity (Ksat). At the end of the incubation, the Ksat values of the amended soils were twice as high as the control soils (Table 2), although there were great variances found at the beginning of the incubation, especially for

the 5% biochar amended DNA Damage inhibitor soil (Fig. 2h). After incubation of 21 d, the Ksat stabilized gradually and kept higher consistently for the biochar-amended soils to the end of the incubation. To understand the changes of soil microbial activity after biochar application, the microbial biomass carbon (MBC) contents were determined at 0 d, 21 d, 63 d, and 105 d of incubation. Results indicate that the biochar application significantly increased the MBC at the beginning of incubation, 63 d and 105 d (only in 5% application rate). The differences were statistically significant (p < 0.05), except for the analytical results at 21 d ( Fig. 3). In addition, the highest contents of MBC were found at 21 d for each treated soil, which were 3200 mg kg− 1 for 5% biochar-amended

soil, 1145 mg kg− 1 for 2.5% biochar-amended soil and 1759 mg kg− 1 for the control, respectively. Table 2 shows the soil loss rate under a simulated rainfall intensity of 80 mm h− 1. The highest soil loss rate (1458 ± 50.0 g m− 2) Rolziracetam occurred in the control soil, and the lowest (532 ± 106 g m− 2) occurred in the amended soil with the highest application rate (5%). The soil loss rate significantly decreased as the biochar application rate increased, indicating that biochar largely ameliorated soil erosion potential in highly weathered soils. The results of this study confirmed the effectiveness of wood biochar in improving the physical and chemical properties of soil that is highly weathered. The results indicated that the improvements in soil characteristics varied with variations in the amount of biochar added to the soil. Incubation results indicated that soil pH, CEC, and BS increased significantly after the addition of biochar, particularly at the application rate of 5%. The high liming potential of the biochar (pH > 9.0) raised the pH of the highly weathered soil. Our results further showed that pH increased significantly with increasing application rates of biochar, reflecting the fact that the liming potential increased with increasing application rates of biochar.

The first year following vaccination, the predicted seroprotectio

The first year following vaccination, the predicted seroprotection rate is high but decreases quite rapidly (−2.3% between day 28 and year 1). The seroprotection rate declines at a slower rate during the second year than during the first (−0.4%) but then accelerates from this point onwards. This can be seen by a steeper curve after year 5. In particular, at year 5 the predicted seroprotection is 94.7% (95% CI: 90.9–97.9) which is comparable

to the observed value of 93.3% (95% CI: 82.1–98.6). At 10 years the predicted seroprotection level still remains high at 85.5% (95% CI: 72.7–94.9). We calculated the percentiles for duration AZD8055 mw of protection in our study population, or equivalently, the percentage of individuals having at least the given duration of protection selleck inhibitor by maintaining antibody titres above the accepted threshold. The maximum, median and minimum duration

of protection were calculated to be respectively 38.1 years, 21.3 years and less than 28 days. Excluding the 2 subjects who were not seroprotected at 28 days (vaccine non responders), all subjects had at least 3.4 years of protection and 90% of subjects had at least 11.2 years of protection. Table 3 gives the percentiles for duration of protection in our study population excluding the 2 non-responders. The change point for antibody decay refers to the time when the initial period of rapid decline in titre ends and the second period of slow decline begins. The average individual change point, as estimated by the 2-period piecewise-linear

aminophylline model, was 0.267 years (5th to 95th percentile range: 0.11–0.61). This means that antibody titres after a single dose of JE-CV would continue to decline rapidly from their peak value observed around day 28 until 3.2 months after vaccination on average (5th to 95th percentile range: 1.4–7.3). After this initial period of rapid antibody decline, titres continue to decline but at a much slower rate (about 50 times slower). Our analyses of the persistence of antibodies predict that the seroprotection rate after a single dose of JE-CV in adults remains high for at least 10 years. This conclusion is based on a median antibody titre at 10 years of 38, which exceeds the seroprotective threshold of 10 accepted by regulatory authorities as a surrogate marker of protection [9]. Overall, we predicted that 85.5% of subjects will maintain antibody titres above the threshold value 10 years after vaccination. The median duration of seroprotection exceeded 20 years, and 90% of responding subjects had at least 11.2 years of protection. We also inferred from our analyses that there is an early, short period of rapid antibody decline ending during the 4th month after vaccination (3.2 months on average), after which a second period of much slower antibody decay ensues for many years.