Both aversive and positive interactions are relevant features of

Both aversive and positive interactions are relevant features of the social environment. Widely used models of social stress in rodents include social subordination, crowding, isolation,

and social instability (Fig. 1, left side). While most studies have been conducted in mice and rats, prairie voles and other social rodent species provide an opportunity to study the role of identity of the social partner, and how separation from a mate differs from isolation from a same-sex peer. In humans, social rejection is used as a potent experimental this website stressor (Kirschbaum et al., 1993), and decades of work in humans and non-human primates have demonstrated that an individual’s position in the social hierarchy has profound implications for

health and well-being (Adler et al., 1994 and Sapolsky, 2005). In rodents, the most prominent selleck chemicals llc model of stressful social interaction is social defeat. Social defeat is typically induced by a version of the resident-intruder test in which a test subject is paired with a dominant resident in its home cage. Dominance may be assured by size, prior history of winning, strain of the resident, and/or prior housing differences (Martinez et al., 1998). Defeat may be acute or repeated, with many possible variations on the method. Social defeat is typically used as a stressor in male rodents, for whom dominance is easier to quantify and aggressive interactions related to home territory are presumed more salient. A few studies report effects of social

defeat on females, particularly in Syrian hamsters in which females are highly aggressive and dominant to males (Payne and Swanson, 1970). In rats and mice, females do not always show a significant response to this task and the effect in males is far greater (Palanza, 2001 and Huhman et al., 2003). Thus, other stress paradigms such as social instability are more widely used with females (Haller et al., 1999). Social defeat can have a more substantial impact on male rodent physiology and behavior than widely used stressors such as restraint, electric shock, and chronic Tryptophan synthase variable mild stress (Koolhaas et al., 1996, Blanchard et al., 1998 and Sgoifo et al., 2014). In the short-term, social defeat produces changes in heart rate, hormone secretion, and body temperature, with longer-term impacts on a wide variety of additional outcomes including activity, social behavior, drug preference, disease susceptibility and others (Martinez et al., 1998, Sgoifo et al., 1999 and Peters et al., 2011). Unlike physical stressors such as restraint, social defeat does not appear to be susceptible to habituation or sensitization (Tornatzky and Miczek, 1993 and Sgoifo et al., 2002), and can be used in groups housed with a single dominant individual (Nyuyki et al., 2012).


these studies are not likely to be a primary strateg


these studies are not likely to be a primary strategy to detect the impact of PCVs and when undertaken are at risk of being confounded by changes in pneumonia burden or mortality trends unrelated to pneumococcal disease (e.g. respiratory viral epidemics, malaria). The assessment of carriage of vaccine type and non-vaccine type pneumococci is a direct, pathogen-specific ZD1839 cost measure of PCV impact that is an indicator of the success or failure of a PCV rollout program [129]. Cross sectional studies of carriage in the target age group of PCV, as well as in older children and adults, will give a measure of herd protection. Detection of important serotypes in developing countries (such as type 1) may still be done in carriage studies if the subjects are carefully chosen, by including the detection of carriage in subjects with pneumonia on arrival at health care facilities. Detection of such rarely carried types in pneumonia patients may reflect an etiological role of those types in pneumonia [137]. Carriage studies focused on young children with respiratory illness will identify the group at risk for pneumococcal disease but also provide access to older siblings who are often transmitters of the pathogen, and mothers who may be key to measurement

of herd protection in adults. Cross sectional studies may detect changes BIBF 1120 in vitro in the distribution of vaccine type carriage as soon as a year post PCV introduction if sample size is sufficient, with detection of profound changes in distribution and herd protection, if present, by 3–4 years post PCV [138]. While carriage studies will not likely be a direct measure of reduction in disease burden due to PCV, they offer a direct measure of program effectiveness and the nature of replacing pathogens, including an assessment of the impact of PCV on the NP microbiome. There are emerging data suggesting that quantitative detection of carriage using microbiological methods,

but also more easily by quantitative PCR, may be diagnostic of pneumonia in adults [139]. These methods may also reflect co-infection with respiratory viruses in children [140] which may be a significant risk for pneumonia hospitalization [141]. The antimicrobial susceptibility profile of carried pneumococci may be used to inform treatment algorithms for pneumococcal disease Unoprostone in developing countries [142]. Quantitative molecular methods may increase the sensitivity of detection of pneumococcal carriage, and may also detect more easily than culture an impact of PCV on density of carriage. The detection of serotypes in carriage can be used together with the global distribution of those types in IPD [143] to develop an invasiveness index that may be predictive of the likelihood of invasive disease replacement due to emerging types detected in carriage. There are advances in work linking the NP and IPD post-PCV impact results, thereby providing a means to predict IPD impact using NP carriage [147].

Asked which vaccines they would most like to see licensed for CTC

Asked which vaccines they would most like to see licensed for CTC use, most vaccinators and supervisors cited other vaccines used in campaigns, with polio (44%), measles (40% and yellow

buy Panobinostat fever (29%) the most commonly cited. Over the course of the campaign, 155,000 people were vaccinated with MenAfriVac in a CTC. This marks the first time since the establishment of EPI that a campaign was conducted using a vaccine with on-label guidance for use beyond the 2–8 °C standard cold chain range. As per the coverage rates attained, the campaign was successful in reaching the target population. The 2013 disease surveillance across Benin—including in the CTC area—supports this, with no cases of Meningitis A reported in the vaccinated population [9]. Cold chain has been a limiting factor since the inception of the EPI. The need to keep vaccines between 2 and 8 °C at all times currently drives the way immunization strategies are developed and implemented. This study provides a first example of the types

of benefits that could be seen from removing that constraint, especially for immunization campaigns and other outreach based strategies. While the rigorous regulatory reviews provided assurance as to the efficacy of the vaccine, the pilot provides critical validation of the acceptability of the practice by health care workers. In addition to the survey results which indicated a strong preference for CTC when feasible, the CTC approach also has the potential to have a positive impact on the provision of

other primary health care initiatives, freeing up health care worker time and resources to learn more keep other regular primary care services operational (often cancelled during Non-specific serine/threonine protein kinase campaigns) [10], rather than managing cold chain and ice pack production logistics [11]. In addition, while the original six EPI vaccines were very sensitive to heat, many new vaccines—including the MenAfriVac diluent—are damaged by exposures to freezing temperatures while remaining stable at higher temperatures for longer periods of time. Studies have shown that freezing is a particular risk during transport and outreach [12]. The CTC practice removes the risk of freezing during these activities at the ‘last mile’. As with any new practice, there were several challenges noted with the CTC implementation. The biggest of these was the need to discard unused vials after four days in a CTC, rather than having the ability to return them to the fridge. This required close supervision by health care workers and district health staff, and if staff are not well trained, could lead to increases in vaccine wastage. Once trained, vaccinators found the peak threshold temperature cards easy to use. However the need to ensure the vaccines are always kept with an indicator provides an additional difficulty, and vial level peak threshold indicators should be considered. Caution must be exercised around storage of the indicator cards prior to use.

The broad peaks at 19 and 38 kDa probably represent monomeric and

The broad peaks at 19 and 38 kDa probably represent monomeric and disulfide-linked forms, respectively, of the M8 VHH coupled to the RS100 array. We observed this artefact more often (results not shown) although not always (Fig. 3). To develop SELDI-TOF-MS analysis of FMDV

antigens we first compared the mass of the spectral peaks found with the predicted mass based on translations of RNA sequences of three FMDV strains. Since the individual MS-275 mw FMDV proteins are cleaved from a polyprotein by the FMDV 3C protease their exact C-termini cannot be deduced from stop codons. We therefore defined VP1-4 termini as done in previous database entries. There is however some controversy about the location of the C-terminus of VP1. Most literature describes VP1 of O1 strains as a protein of 213 amino acids ending with amino see more acid sequence KQLL or KQTL without relying on experimental data. Examples are Refs. [14] and [17]. However, such cleavage is not consistent with cleavage of the peptide APAKQLLDFDLLK by 3C protease after a glutamine (Q) residue [18], nor with identification of the 2A peptide located C-terminal from VP1 as LLNFDLLKLAGDVESNPG [19]. Thus, we defined the VP1 C-terminus as ending at KQ, resulting in a protein 2 amino acids shorter than previous definitions. We will now discuss the identification of the different peaks

of strain O1 Manisa separately. Since VP4 is myristoylated [15] the peak at 9.0 kDa must represent myristoylated VP4. The identification of this peak as VP4 is confirmed by its absence in SELDI-TOF-MS experiments where 12S particles generated by acid treatment were captured by M8 (results not shown) since 12S particles are known to lack VP4 [2]. The VP4 peak is also observed in SELDI-TOF-MS experiments where untreated FMDV antigen Unoprostone was captured by the M8 or M23 VHHs,

but not using the M3 VHH. This indicates that M3 specifically binds 12S particles. This interpretation is consistent with the previous observation that M8 and M23 do but M3 does not neutralize FMDV in vitro [13]. A closer view showed that VP4 actually consists of 8 peaks with a 14–17 Da difference. This could represent different degrees of oxidation of VP4, which results in 16-Da differences. Oxidation is a modification of Met, Tyr, Trp, His and Cys residues that occurs easily during protein storage [20]. Surprisingly such heterogeneity is only observed with VP4 but not at all with VP1-3. Since only VP4 contains a myristoyl group this could indicate involvement of this group with the observed heterogeneity, possibly due to oxidation of this group. VP1 occurred as two variants of 23.3 and 23.5 kDa. Their identification as VP1 is confirmed by their abolishment by trypsin treatment which cleaves only VP1 without dissociation of 146S particles [17] and [21]. The origin of the VP1 heterogeneity is unclear.

The Clark scale is a 24-point scale based on duration and frequen

The Clark scale is a 24-point scale based on duration and frequency of diarrhea and vomiting, degree and duration of fever measured by rectal temperature, and description and duration of behavioral symptoms. Axillary temperature measurements were used instead of rectal measurements. Conversion of axillary temperature to rectal temperature was performed using following formula [7]: rectal temperature (°C) = 0.98 × axillary temperature (°C) + 0.8 (°C). The Clark scale is divided into three ranges: mild <9, moderate 9–16, and severe >16. The Vesikari scale is a 20-point scale based on duration and peak frequency of diarrhea and vomiting, degree

of temperature, severity of dehydration, and treatment provided to the patient (i.e., rehydration or hospitalization). This scale is divided into three ranges: mild <7, moderate 7–10, and severe ≥11 [9] and [10]. Stool sample (1.5–5 g) was collected for each subject, preferably at enrollment, or later Crizotinib order but within 14 days of the onset of AGE symptoms. The stool samples were stored at 2–8 °C. Samples were shipped to The Wellcome Trust Research Laboratory

(Department of Gastrointestinal Sciences, Christian Medical College, Vellore, Tamil Nadu), which was the central laboratory for this study. The samples were shipped in batches and laboratory testing occurred after the 14 days follows up of individual subject was over. Thus, the investigators or the site staff was not aware if subject was suffering from RVGE or non-RVGE when AGE related data was collected progestogen antagonist Dipeptidyl peptidase and severity scoring was done. Stool samples were first tested for the presence of rotavirus antigen by enzyme immune assay (EIA) using Prospect™ Rotavirus EIA. The samples that were positive by EIA were genotyped for their respective G and P types by RT-PCR. For RT-PCR, viral DNA was extracted from stool specimens and reverse transcribed using random primers to generate complementary DNA (cDNA). The cDNA was used as a template for genotyping in hemi-nested multiplex PCRs for VP7 and VP4 genes using published primers and protocols [10], [11], [12], [13] and [14]. The primers

could amplify VP7 genotypes: G1, G2, G3, G4, G8, G9, G10, and G12; and VP4 genotypes: P[4], P[6], P[8], P[9], P[10], and P[11]. The study was conducted in accordance with the ethical principles enshrined in the Declaration of Helsinki, International Conference on Harmonization (ICH) – Guideline for Good Clinical Practice (GCP), and all applicable local regulatory requirements. The study protocol was approved by the Ethics Committees for respective sites. Per protocol (PP) population was used to analyze the study data. Subjects who had a total data of 14 days, EIA results available, and completed the study as per protocol were included in the PP population. The proportion of RVGE among AGE was calculated for regions and overall (with 95% CI). Data were summarized using number and percentages, mean, median and other statistics as appropriate.

Dextrose solution was transfused continuously throughout the peri

Dextrose solution was transfused continuously throughout the period of study. Periodically, 1 ml of blood sample was taken by syringe containing 1 ml of heparin solution to prevent blood clotting. These blood samples were centrifuged at 2500 rpm for about 30 min. One milliliter of the supernatant was taken, and after suitable dilution, analyzed at 362 nm spectrophotometrically by the method described under in vitro analysis. The optimized formulations (AF4 and AT5) were selected and the stability studies were carried out at accelerated condition

of 40 ± 2 °C, 75 ± 5% RH conditions, stored in desiccators, the formulations were packed in amber color screw cap container and kept in above-said condition for period of 3 months. The formulations were analyzed periodically for their physical appearance, buccoadhesive

strength and in vitro drug release. The FTIR spectra of Amiloride hydrochloride, HPMC, MLN8237 cell line SCMC, Eudragit, Carbopol, Chitosan and PVP and the combination of drug and polymers showed no significant interaction between drug and polymer. The spectral data of pure drug and various drug-excipient mixtures are tested. The results indicate that there was no chemical incompatibility between drug and excipients used in the formulation. The surface pH of the formulations was determined in order to find out the possibility of any side effects in buccal environment. The observed surface pH of the formulations was found to be in the range of 5.82–6.52. The results shown that there also is no significant difference in the surface pH of all the formulations and the pH range lies within the range of salivary pH, i.e. 6.5–6.8, thereby not causing irritation in the SB203580 site of administration. Buccoadhesive strength of buccal films is shown in Fig. 1 and swelling index of buccal tablets is shown in

Fig. 2. The stability study of the optimized formulation was done in natural human saliva. The films did not exhibit any significant changes in their color, shape and had satisfactory physical stability. Carbopol, being an anionic polymer, gives the highest buccoadhesive force. The buccoadhesive strength exhibited by Amiloride hydrochloride buccal films was satisfactory for maintaining them in oral cavity. The combination of HPMC and CP shows good adhesion. Upon addition of PVP, the buccoadhesive strength increases which may be due to hydrogen bond formation and Vander Waals forces. Swelling of buccal tablets at different time intervals shown in Fig. 3. Data of in vitro release were fit into different equations and kinetic models to explain the release kinetics of Amiloride hydrochloride from the buccal tablets. The kinetic models used were a zero-order equation, Higuchi’s model and Peppa’s models. The obtained results in these formulations were plotted in various model treatments as cumulative percentage release of drug versus square root of time (Higuchi’s) and log cumulative percentage release versus log time (Peppas).

The full MERS-CoV genome isolated from a Qatari dromedary camel i

The full MERS-CoV genome isolated from a Qatari dromedary camel is highly similar to the human England/Qatar 1 virus isolated in 2012 and has efficiently been replicated in human cells using human DPP4 as entry receptor, providing further evidence for the

zoonotic potential of dromedary MERS-CoV [10]. Although, we cannot conclude whether the people were infected by camels or vice versa or if yet another source was responsible, increasing evidence indicates that camels I BET151 represent an important link in human infections with MERS-CoV. Intensive vaccine control and risk-reduction targeting dromedary camels might be effective in eliminating the virus from the human population. The coronavirus spike protein (S) is a class I fusion protein. Cellular entry of the virus has been demonstrated to be mediated by the S protein through the receptor binding domain (RBD) in the N-terminal subunit (S1) and the fusion peptide in the C-terminal subunit (S2) [11] and [12]. For betacoronaviruses, the S protein has been shown to be the main antigenic component responsible for inducing high titers of neutralizing antibodies and/or protective immunity against

infection in patients who had recovered from SARS [13] and [14] and response levels correlated well with disease outcomes [15] and [16]. The S protein has therefore been selected as an important target for vaccine development [17], [18], [19], [20] and [21]. Recent work shows that modified vaccinia virus find more Ankara expressing the S protein of MERS-CoV elicits high titers of S-specific neutralizing antibodies in mice [22]. Adenovirus 5 (Ad5)-vectored

candidate vaccines induce potent and protective immune responses against several pathogens in humans and a variety of animals [18], [23], [24], [25], [26], [27], [28], [29], [30], [31], [32] and [33]. Although a trial of a candidate DNA/rAd5 HIV-1 preventive vaccine showed lack of efficacy [37] and the high prevalence of pre-existing anti-Ad5 immunity may have been a major limitation [38] in humans, replication-defective adenovirus vaccines are among the most attractive vectors for veterinary vaccine development, given the relative speed and low cost of development and production. Most adenoviruses infect their host through the airway epithelium and replicate in the mucosal tissues of the Linifanib (ABT-869) respiratory tracts [39]. Because of their ability of to elicit mucosal immune responses, adenoviruses could be an attractive vector for inducing MERS-CoV-specific immunity in dromedary camels, the putative animal reservoir. Interestingly, sera antibodies against adenovirus type 3 were detected in 1.3% of dromedaries in Nigeria [34] and in 43 of 120 camels in Egypt [35]. The occurrence of adenovirus type 3 respiratory infections in camels was studied in Sudan and a 90% seroprevalence was detected [36]. Here, we describe the development of recombinant type 5 adenoviral vector expressing, codon-optimized MERS-S and MERS-S1 (Ad5.

(n = 15), or an unknown reason (n = 34) Refusals were included a

(n = 15), or an unknown reason (n = 34). Refusals were included and coded as limited health literacy, as these people are likely to perform with limited health literacy skills in real-life settings (e.g. at the doctor’s office) because of their difficulties. Therefore, they were included to maintain the population-representativeness

of the sample and capture a more accurate range of the health literacy skills of the English population. The present analysis thus included 3087 men and women aged 60–75 years (Fig. 1). Health literacy was assessed using a four-item comprehension test based on a fictitious medicine label from the International Adult Literacy Survey (Thorn, Anticancer Compound Library cost 2009) (Appendix A). Health literacy was categorised as ‘adequate’ (4/4 questions answered correctly) or ‘limited’ (< 4/4 answered correctly) to capture the point at which adults begin to have difficulty with everyday health tasks. Although whether and how health literacy skills may change over time are uncertain, health literacy scores among our sample

are expected to be stable between data collection and the times of reported CRC screenings (within one year of wave 5 data collection for 59% of those reporting screening and within two years for 96%). Health literacy was also measured at ELSA wave 2 (2004–5) and the scores did not change between waves 2 and 5 within individuals who remained in the study for both waves. Health literacy scores measured Kinase Inhibitor Library purchase at wave 2 were not used for this analysis, as study attrition between waves was differential by health literacy score. Participants were asked if they had ever used a bowel testing kit (i.e. an FOBT kit) and whether the kit was part of the NHS Bowel Cancer Screening Programme. Only 49 out of the 1709 participants (< 3%) who reported having completed an FOBT kit responded that the kit was not part of the NHS programme

and 3 (< 1%) responded that they did not know whether it was part of the programme; hence for this analysis we assume that completion of a Oxymatrine FOBT kit equates with participation in the NHS programme. For convenience, the terms “completion of an FOBT kit” and “CRC screening” will hereupon be used synonymously. Sociodemographic covariates were: age, sex (male; female); educational attainment (no qualification; up to degree level; degree level or equivalent); net non-pension wealth (quintiles stratified at age 65 to account for changes in wealth following retirement) (Bostock and Steptoe, 2012); occupational class according to the 2010 National Statistics Socio-economic Classification (routine; intermediate; managerial or professional) (Office for National Statistics, 2010); and ethnic minority status (non-white; white).

55 (d, 1H, 3H, J = 1 8), 6 25 (s, 2H, 7 amino), 5 5 (s, 2H, -CH2-

55 (d, 1H, 3H, J = 1.8), 6.25 (s, 2H, 7 amino), 5.5 (s, 2H, -CH2-NCS), 4.48 (s, 2H, N-CH2). Solution of 35 mg (0.1 mmol) of DTPA dianhydride in 0.3 ml of DMSO obtained under heating to 60–80 °C was cooled down to room temperature and added to 20 mg (0.048 mmol) of compound III. The reaction was carried on for 15 min at 20 °C. The mixture was supplemented with 4 ml of water, left for 20 min at room temperature and pH was adjusted to 5.0 by LiOH. The product was purified by preparative C-18 HPLC column (20 × 250 mm) using linear gradient (0.5l) of acetonitrile in water (0–70%). The elution rate was 2 ml/min. The fractions containing desired product selleck inhibitor were combined and supplemented

with one equivalent of a lanthanide salt. The resulting solutions were concentrated Panobinostat mw in vacuo by co-evaporation with acetonitrile under gentle heating (25–30 °C) to final concentration 20 mM. The reaction cocktails (10–16 μl) were composed by mixing of 7 μl of avidin (20 mg/ml), 1 μl of 1 M sodium borate buffer pH 10.0, and 1–8 μl of a reactive light-emitting probe at concentrations specified in figure legends. After incubation for 4 h at 56 °C the mixtures were diluted to 100 μl by water and subjected to size-exclusion chromatography on Sephadex G-50 “medium” in

10 mM Hepes-HCl buffer pH 8.0 containing 50 mM NaCl. The fractions corresponding to modified avidin were collected by visual detection using UV monitor (365 nm light). LB broth (100 ml) was inoculated with suspension of 10 μl of E. coli cells (RL721 strain) and incubated in a 500 ml Erlenmeyer flask overnight at 37 °C. The cells were harvested by centrifugation (4000 rpm, 5 min), washed with PBS and re-suspended in the

same buffer containing 50% glycerol at a final density of 32 mg ml−1. Thirty microliters of this suspension containing ca. 1 mg of cells was washed 3 times with 1 ml of 0.1 M sodium borate buffer, pH 8.5, and each time collected by centrifugation. After the last wash, Calpain the cells were suspended in 50 μl of the same buffer and 4 μl of 100 mM DMSO solution of NHS-dPEG12-biotin was added. After incubation at room temperature for 30 min the cells were washed 4 times with 500 μl of PBS. After the final wash, cells were suspended in 15 μl of PBS buffer and supplemented with 15 μl of 5 μM avidin modified with one of the lanthanide labels [AV – Probe 4 -Tb3+ (n = 15) and AV – Probe 1-Eu3+ (n = 19)]. After 25 min of the incubation at room temperature cells were washed by PBS (4 × 500 μl) and suspended in 100 μl of the same buffer. CHO cells were grown in Dulbecco’s modified Eagle’s medium, supplemented with 10% fetal bovine serum, 200 mM l-glutamine and 100 g/ml penicillin/streptomycin solution. Once the cells reach 80–90% confluency, they were trypsinized and collected by centrifugation (1000 rpm for 5 min), washed with 0.1 M Na-borate buffer pH 8.5 (3 × 0.5 ml) and spun down at 3000 rpm for 30 s.

The same precautions still need to be taken, and all the surveill

The same precautions still need to be taken, and all the surveillance, but the number of people who are actually suffering from the disease will be very small. It is at this point that vaccine refusal is more likely to become a problem – as individuals may not unreasonably question whether they themselves stand to benefit from the vaccination. Where policymakers take the view that eradication should continue to be pursued only where the cost-per-QALY for each individual case remains within tolerable bounds, then they are likely to give up before the job has finished Dorsomorphin – meaning that there will be continued flare-ups of the disease, with the net result that the disease will never be eradicated

[21]. Third, and most difficult, there is a deep question about how to weigh even successful eradication campaigns in the balance against other uses of healthcare resources. Disease eradication brings its true benefits only over the long term, whilst healthcare spending tends to focus on short to medium-term benefits. If we assume that it is equally as important to save a life in fifty A-1210477 ic50 or a hundred years’ time as it is to

save one now, then it would seem that we should devote a very great proportion of our current healthcare resources to eradication campaigns. As Murray [22] put this point in setting out the initial framework for the Global Burden of disease report: if health benefits are not discounted, then we may conclude that 100% of resources should be invested in any disease eradication plans with finite costs as this over will eliminate infinite streams of DALYs which will outweigh all other health investments that do not result in eradication. Murray drew the conclusion that in order to avoid this paradox, future health benefits should be subject to a discount rate. This conclusion seems surprising: if the expected

total health benefits of eradicating a disease such as malaria really were vastly greater than, say improving control of diabetes, would not this be a strong argument in favour of eradication? Whilst the terrain here is complex, there seems to be no good reason to apply large discount rates to future health benefits, even if there are good reasons for significantly discounting other future goods [23]. It is standard in economics to apply a discount rate to commodities, because the price of most commodities falls over time relative to the return we could get on an investment at a bank. This discounting model assumes that the increased amount of commodities that could be bought in the future with the money invested has the same value for wellbeing as the smaller bundle we can buy now. However health gains and avoidance of death would seem to contribute a constant amount to wellbeing whenever they occur. So these reasons for discounting commodities do not imply that future health should be discounted [24]. Economists also argue in favour of a discount rate on the grounds of uncertainty.