Sometimes WHO representatives may also participate in the working

Sometimes WHO representatives may also participate in the working

groups. After assessing all available data, the committee will reach consensus and recommendations will be made. If consensus proves impossible, the matter will be sent to the MoH, to make the final decision. Agreed Target Selective Inhibitor Library recommendations are forwarded to the ultimate decision-makers within the MoH and then widely circulated via circulars and newsletters. It should be noted that to date the committee has always followed official WHO recommendations for vaccine use. Formal contact between the committee members and similar NITAGs in the Gulf Cooperation Council (GCC) countries is facilitated through an annual inter-country Selleckchem Dolutegravir meeting on communicable diseases that includes all the countries of the GCC. This comprises Bahrain, Kuwait, Oman, Qatar, Saudi Arabia, the United Arab

Emirates and Yemen. Half of the meeting each year is devoted to discussing issues concerning the Expanded Programme on Immunization (EPI), including the introduction of new vaccines and immunization issues. In 2007, it was recommended that GCC countries would have a common EPI schedule, a decision validated by all NITAGs in GCC countries and then approved by the relevant Ministers. As of 1 January 2008, the decision was implemented. The cost of vaccines, as well as that of the overall immunization program, is considered when the committee decides on its recommendations. Formal economic evaluations are made (cost-effectiveness, cost-benefit and cost-utility) and both affordability and sustainability are assessed. Subcommittees, with the assistance of health economic experts from within the MoH, assist in making these evaluations—for example, an economic evaluation of rotavirus vaccine disease burden was undertaken. They are currently assessing the human papillomavirus (HPV) disease burden from an economic

perspective. Additionally, assessments made regionally are taken into account, particularly when provided by WHO’s Eastern Mediterranean Regional Office (EMRO) or from other GCC countries, such as in the case of cost-effectiveness studies on HPV. Recommendations are circulated to all members to receive their comments, after which they are sent those to decision-makers for final approval. The Government is obliged to implement committee recommendations. The Ministry of Finance and other government departments play no part in decision-making. A good example of how decisions are made can be found in the case of the introduction of PCV-7 into the EPI schedule in Oman. At the time, there was very strong demand from the vaccine committee members and paediatricians to introduce the vaccine. As a result, the committee recommended forming a task force to study the disease burden and the vaccine’s cost-effectiveness.

1a) Because of this porosity, higher amounts of biochar in the t

1a). Because of this porosity, higher amounts of biochar in the treated soil increased the habitat for microbes to grow. Joseph et al. (2010) indicated that most of biochar has a high concentration of macro-pores that extends from the surface to the interior, and Selleckchem Androgen Receptor Antagonist minerals and small organic particles might accumulate in these pores. Few studies have been published

on the influences of biochar on the physical properties of soils (Atkinson et al., 2010). In addition to improved chemical properties of the soils, our results indicated a particularly significant improvement in the physical properties of the highly weathered soil. The results indicated a significant decrease in Bd, and an increase in porosity, Ksat, and the MWD of soil aggregates in the biochar-amended soils, even at the low application rate (2.5%) after incubation of 105 d (Table 2). During the incubation duration, the values of Bd kept higher in the biochar-amended soils 3-MA nmr than in the control after 21 d. Before 21 d, the rapid increase

in the control’s Bd might be caused by gradual infilling of clays into pores of the soil, which reflected that the incubated soils are stable and approached field condition after 21 d. For the biochar-amended soils, physical dilution effects might have caused reduced Bd levels, which agreed with Busscher et al. (2011) who indicated that increasing total organic carbon by the addition of organic amendments in soils could significantly decrease Bd. Furthermore, the decrease in Bd of the biochar-amended soils appears to have also been the result of alteration of soil aggregate sizes, as shown by Tejada and Gonzalez (2007) who amended the following soils by using organic Idoxuridine amendments in Spain. In our study, micromorphological observations of the amended soils indicated the flocculation of soil microaggregates after the addition of biochar (Fig. 4a; b). The porosity could also be effectively improved by application of the biochar and hydraulic conductivity as well.

Asai et al. (2009) indicated that the incorporation of biochar into rice-growing soils changed the pore-size distribution, which increased water permeability. Regarding the porosity and hydraulic conductivity of the amended soils, we considered the redistribution of the proportion of soil aggregate sizes to be a critical factor in influencing the physical and chemical properties of the soil (Table 2). The incorporated biochar could function as a binding agent that connects soil microaggregates to form macroaggregates. The oxidized biochar surface, which included hydroxyl groups and carboxylic groups, could adsorb soil particles and clays (Fig. 4c) to form macroaggregates under acidic environments. Our incubation study showed that the biochar-amended soils seemed to have larger soil aggregates than the control after 21 d although significant difference of MWD was just found after 63 d between the amended soils and the control.

The friability of

tablets was measured in Roche friabilat

The friability of

tablets was measured in Roche friabilator. Twenty tablets were dedusted at 25 rpm for 4 min and weighed again.8 Percentage friability was calculated from loss in weight as given in equation below. The weight loss should not be more than 1%. %Friability=[(Initialweight−Finalweight)/Initialweight]×100 The test was carried out on 6 tablets Ku-0059436 manufacturer using digital tablet disintegration test apparatus (Microprocess based-Electrolab). Distilled water at 37 °C ± 2 °C was used as a disintegration media and time in second taken for complete disintegration of tablet with no residue remaining in apparatus.10 Percent drug release of venlafaxine hydrochloride mouth dissolving tablets was determined by USP dissolution test apparatus (Lab India − 2000) using paddle method. The dissolution test was performed using 900 ml of phosphate buffer pH 6.8 at 37 °C ± 0.5 °C at 50 rpm. A sample of 5 ml solution was withdrawn from dissolution apparatus at regular interval of 30 s. The same quantity of sample was replaced with fresh dissolution medium. The samples were filtered through 0.45 μm membrane filter.11 and 12 Absorbance of these samples was analyzed at λmax 225 nm using UV–visible spectrophotometer. Wetting time of tablets can be measured using simple procedure. Six circular tissue papers of 10 cm diameter were placed in a petridish.

10 ml of water containing amaranth dye was added to petridish. A tablet was carefully placed on the surface of tissue paper.10 Time required for water to reach upper surface of the tablet was noted as wetting time. The porosity of tablets was calculated from the weight of tablet (W), tablet volume (V), and true density of powder (ρ) using following equation, 13 %Porosity=[1−weightoftablet(W)/Volume(V)×Density(ρ)] The true density of powder was determined by a pycnometer. Photomicroscope (Olympus cx31) was used for pore analysis. FTIR spectra of all formulations were obtained on IR-spectrophotometer (Prestige-21-shimadzu). The samples were prepared in KBr dish (2 mg of sample in 200 mg KBr). The sample scanning range was 500–4000 cm−1. The

surface morphology of optimized formulation before and after Adenosine sublimation of camphor was studied using (GEOL Ltd. Japan-JSM-6360). The tablet surface was sputter coated for 10 min with gold by using fine coat ion sputter and examined under SEM. The stability of optimized formulation F3 was tested according to ICH guideline, at 40 °C ± 2 °C/75%RH ± 5% condition in stability camber (HMG, India) for 3 months.14 Tablets were tested for drug content for 30, 60, and 90 days. The 32 factorial design was used for the optimization of mouth dissolving tablets of venlafaxine hydrochloride (Design Expert The two independent factors, concentration of Indion-234 (X1) and concentration of camphor (X2), were set to three different levels and experimental trials were performed for all nine possible combinations.

03, 95% CI 0 58 to 1 84) This randomised controlled trial examin

03, 95% CI 0.58 to 1.84). This randomised controlled trial examined the benefits and harms of neural tissue management as an intervention for nerve-related neck and arm pain. Low NNTs and moderate standardised mean differences show that neural tissue management produced clinically important benefits for participant-reported improvement, pain intensity, and activity limitations at short-term follow-up when compared to advice to remain active. There was no evidence to suggest that neural tissue management was harmful. The prevalence of worsening was similar for the experimental and control groups, and

no participants had to stop neural tissue management early because of an exacerbation that they and the physiotherapist related phosphatase inhibitor library to treatment. Although several participants experienced adverse events that they related to neural tissue management, these events would be categorised as ‘mild’ because they did not require additional treatment, usually lasted < 24 hours, had minimal impact on daily activities, and did not reduce a participant's chance of improving with neural tissue management (Carlesso et al 2011, Carnes et al 2010). The proportion of participants assigned to neural tissue management

who experienced an adverse event and the characteristics of these events are similar to those reported previously for manual therapy for patients with neck pain Selleck Ruxolitinib (Hurwitz et al 2004). The results of this trial enable physiotherapists to have informed discussions with patients about the short-term benefits and harms of neural tissue management for nerve-related neck and arm pain. Standardised mean differences for pain were similar to results from the trial by Allison and colleagues (2002) (≥ 0.7 versus 0.71), while those for activity limitations were larger (≥ 0.6 versus 0.34) (Gross et al 2004). The consistently favourable results for neural tissue management support the hypothesis that the lack of statistical significance in this previous trial was due to the

small sample.limitations of our study. Time constraints The size and source of the sample, comparison to advice to remain active, and short-term second follow-up are potential limitations of our study. Time constraints prevented enrolment of the a priori sample of 84 participants. Although we anticipated that approximately 10% of volunteers would enter the trial, the response to each recruitment advertisement was lower than expected. Enrolment stopped at 60 participants because data collection could not extend beyond two years. The concern with early stoppage of a trial is that any treatment effect may reflect a ‘random high’ in the data rather than the ‘true’ effect ( Moher et al 2010).

0 1, 0 25 and 0 5 mA/cm2 current densities were used as variable

0.1, 0.25 and 0.5 mA/cm2 current densities were used as variable condition in Iontophoresis while keeping Selleckchem Panobinostat current pattern as continuous DC current. DTAB micellar solution containing Lovastatin in phosphate buffer pH 7.4 was charged in donor compartment of modified Glickfeld diffusion cell. In one experiment, 0.5 mA/cm2 DC current source was kept in continuous mode and in the other

experiment it was kept in 10 s on/off (pulsed) mode. Ten to twelve week old male albino rats (250 g) were sacrificed by excess of ether inhalation. After removing hairs, full-thickness of rat abdomen skin was surgically removed. The rat epidermis was isolated by a heat separation technique and carefully cleaned with normal saline. Finally fat tissue adhered to skin removed by wiping it with cotton swab soaked in isopropyl alcohol and dried under the vacuum followed by storing in desiccators.7, 8 and 9 Skin samples were used within three days of isolation.

Protocols for the use of animal for the above experiment was previously approved from the Institutional animal ethics committee, Noble Group of Institutions, Junagadh. Iontophoresis experiments were carried out at 37 ± 2 °C. All analytical works for quantification Gemcitabine supplier of Lovastatin were done by validated RP-HPLC analytical method by using 0.1% phosphoric acid solution and acetonitrile (65:35 v/v) as mobile phase. Selected composition was charged for stability

study under accelerated stability study condition as per ICH guideline. Selected composition was studied for Zeta potential determination, pH and assay of Lovastatin and in-vitro permeation rate. DTAB was selected as a surfactant for composition for Iontophoresis experiments because single surfactant micelle possesses best solubilizing power than mixers of surfactants specially in context of micellar solubilization of drugs.10 Solubility of Lovastatin was found to be 0.1 mg in 3.7 × 10−3 mol/L of DTAB which is more than 230 folds generally observed in purified water. Fig. 1 show CMC of DTAB in 0.1 mg Lovastatin containing solution under various temperature conditions and it was evidenced that the maximum shift of CMC was up to 3.87 × 10−3 mol/L at ADAMTS5 40 °C. So, use of 3.87 × 10−3 mol/L DTAB in composition can keep Lovastatin in soluble form in core of liquid crystals formed by micelles of DTAB. Passive diffusion of Lovastatin allowed 3.63 ± 0.10 μg/cm2/h Lovastatin permeation rate after 12 h Iontophoresis with 44.36 ± 4.02 μg/cm2 cumulative permeation of drug. Phosphate buffer pH 7.4 as vehicle system provided highest drug permeation with Permeation Enhancement Ratio (E.R.) 1.80 in comparison of passive diffusion (Table 2) (Fig. 2). Lesser E.R. was observed in case of NaCl containing solution may be due to counter ion effect produced by Cl− of NaCl on DTAB micelles.

All statistical calculations were performed using Stata version 8

All statistical calculations were performed using Stata version 8.0 (College Station, Texas, Stata Corporation, 2003). Of the original sample of 1670 physicians, 120 were ineligible because they were retired or no longer in clinical practice. The final sample size included 1550 physicians, of which 1079 responded (overall response rate: 69.6%). Responders and non-responders were comparable in terms of demographic characteristics (location, gender, and age; p > 0.05). Most responding physicians were from Rome (73.8% of responders vs. 76.9% of non-responders) and male (56.2% of responders vs. 58.9% of non-responders), with a mean age of 50.7 (± 11.5) years (50.0 years Afatinib for non-responders).

The demographic characteristics of the sample were similar to those of all KPT-330 mw Italian physicians, as 60.6% of the members of the National Board of Physicians are male and have a similar age distribution ( ENPAM, 2012). Other demographics,

professional and personal characteristics of the responding physicians are listed in Table 1. Italian physicians’ knowledge of predictive genetic testing for cancer appeared adequate in terms of BRCA1/BRCA2 testing, although knowledge of APC testing was lacking [ Table 2(A)]. Almost half of the sample (42.8%) answered all three questions about BRCA1/2 testing correctly. This knowledge was improved if physicians were exposed to cancer genetic testing during graduate or postgraduate training, and with the increase in the amount of time dedicated to continuing medical education. Metalloexopeptidase Female physicians were more likely to have adequate knowledge about BRCA1/2 testing, and this knowledge increased if genetic testing laboratories were located in the same geographical area as the physicians’ workplace (Model 1 in Table 3). Only 16.9% of physicians provided correct answers to all three questions about APC testing. This knowledge, as in the previous case, increased with exposure to cancer genetic testing during graduate and post-graduate training and with the amount of time dedicated to

continuing medical education (Model 2 in Table 3). Physicians’ knowledge was satisfactory on the penetrance of BRCA1/BRCA2 mutations, but not regarding the prevalence of hereditary breast cancer. Most physicians knew that the absolute risk of developing breast cancer in the presence of BRCA1/BRCA2 mutations is 40–80%, but less than one third recognized that the percentage of breast cancer cases associated with BRCA1/BRCA2 mutations is 1–10% [ Table 2(B)]. By contrast, knowledge concerning inherited forms of colorectal cancer was inadequate, as none of the surveyed physicians knew that the percentage of colorectal cancer cases associated with APC mutations is less than 5%, and only a small proportion of physicians recognized that the absolute risk of developing cancer in the presence of APC mutations is 100% [ Table 2(B)]. Attitudes toward predictive genetic testing for breast and colorectal cancer were quite heterogeneous (Table 4).

The lymphoma risk, especially the primary CNS lymphomas, has sign

The lymphoma risk, especially the primary CNS lymphomas, has significantly decreased, in the era of combination anti-retroviral therapy (cART) [7]. An

over-risk for non-Hodgkin lymphomas (NHL) still remains in HIV-infected patients [8] and [9]. Defective T-cell immunity in patients has previously been shown to result in an abnormally high number of EBV-infected B cells in blood, e.g. in chronic active EBV infection, post-transplant patients and in HIV-infected patients MAPK Inhibitor Library high throughput [10] and [11]. Vaccination including adjuvant may affect the EBV-host balance, especially in immunocompromised individuals, e.g. those with HIV-1 infection as it affects HIV-1-, EBV-, CMV- and/or HCV-specific CD4 T-cells [12] and [13]. This vaccination effect on specific CD4 T-cells might in turn also affect the B-cell compartment [14] and [15]. Trichostatin A A history of symptomatic primary HIV-1 infection (PHI) is also known to affect the composition of the B-lymphocyte pool [16]. In this paper we analyse subgroups of non- or

insufficiently antiretroviral treated HIV-infected patients and their EBV-host relation measured by EBV genome load. Vaccination with recombinant HIV-1 gp160 (rgp160)/adjuvant and symptomatic primary HIV-infection (PHI) both affects B-cell function. We show an increased EBV-load in blood B-cells after therapeutic vaccination and a further enhancement of EBV-DNA in patients with a history of PHI. Sixty HIV-1 positive patients from the outpatient clinic at Huddinge hospital and/or South Hospital, Stockholm, including 42 participants in vaccine trials were randomly selected for this study (Table 1 and Table 2). After informed consent 20 mL of blood was collected. HIV-1 negative controls (not matched for age, sex or risk group) were selected among healthy laboratory personnel. Permission for the study was obtained from the regional Ethical Committee at the Karolinska Institute (#51/97). Of the 42 immunised individuals, 32 participated for two years in a double blind placebo controlled phase III vaccine trial with r160 (rgp160)/”placebo” [17]. Both

in the rgp160 vaccine and placebo arm alum was included as adjuvant. In this early vaccine trial patients received at least eight vaccinations with alum/rgp160 Non-specific serine/threonine protein kinase at regular intervals for 21 months. Placebo was given according to the same time schedules. The other 10 patients were during more than three years included in an on-going open phase II clinical study with the same vaccine. These patients got 12–16 vaccinations during three years. The patients were treated during the pre-HAART/cART era with one or two nucleoside analogues or foscarnet. We designate this treatment regimen as insufficient antiretroviral therapy, as indicated both by CD4 counts and breakthroughs of HIV RNA levels. Sex, age, patient origin, route of transmission, CD4+ and CD8+ cell counts are summarised in Table 1.

For the third experiment (experiment 3) carried out at Anses Plou

For the third experiment (experiment 3) carried out at Anses Ploufragan, France, Large White pigs were obtained from a local high health status farm and the average weight at the

first immunisation was 11 kg. All pigs were maintained at high security facilities throughout the experiment. The first experiment at Pirbright was performed under Home Office licence PPL 70-6369. Experiments at Ploufragan were performed according to the animal welfare experimentation agreement given by the Direction des Services Vétérinaires des Côtes d’Armor (AFSSA registration number B-22-745-1), under the responsibility of Marie-Frédérique NVP-BKM120 price Le Potier (agreement number 22-17). Briefly, pigs were intramuscularly inoculated

with 104 TCID50 of non-virulent ASFV isolate OURT88/3 and boosted intramuscularly 3 weeks later with 104 HAD50 of virulent ASFV Anti-infection Compound Library isolate of OURT88/1. Pigs were then challenged 3 weeks later with 104 HAD50 of either Benin 97/1 or virulent Uganda 1965 intramuscularly. ASFV-inoculated pigs were monitored for body temperature and other clinical symptoms and these were recorded and scored according to the clinical scoring system shown in Supplementary Table 1. Weight gain was also recorded in the experiments carried out at Ploufragan. All pigs were examined post-mortem either when the pigs died or at the termination of the experiments. Tissues were collected for further analysis. Peripheral blood was analysed at different days post-immunisation for the presence of ASFV by quantitative PCR (qPCR) as described previously [22]. Samples which tested positive by qPCR were further analysed by cytopathic and/or haemadsoption assay (HAD) using standard pig bone marrow cells in 96 well plate [23] and [24]. Spleen, tonsil, retropharyngeal and ileocaesal 17-DMAG (Alvespimycin) HCl lymph nodes from post-mortem tissues were also analysed for the presence of ASFV by qPCR and HAD. Virus detected from tissue samples

by qPCR was expressed as copy number per mg tissue and by HAD as HAD50. Development of T cell immune responses to ASFV after immunisation was analysed by IFN-γ ELISPOT and proliferation assays as described previously [25]. All ASFV isolates used as antigens for T cell assays were prepared by culture in porcine bone marrow cells, and ASFV titres were determined by qPCR [22] and adjusted to give the equivalent of 105 HAD50/ml. Uninfected porcine bone marrow culture supernatants were used as negative control antigen. The development of ASFV specific antibodies was analysed using a competition ASF ELISA kit (INGENASA PPA3 COMPPAC), and the antibody titre was expressed as log 2 dilution of end point which gives 50% competition.

4 per 1000 child-years (95% CI, 87 2, 97 9) The use of these bro

4 per 1000 child-years (95% CI, 87.2, 97.9). The use of these broad criteria for active surveillance resulted in many children with non-specific illness being screened at a hospital and undergoing an ultrasound examination. The screening protocol resulted in only 1.6% of the possible cases being classified as PI3K inhibitor ultrasound-evidenced intussusception and 0.8% Brighton level 1 confirmed intussusception. Based on this study, the broad screening approach met the safety criterion of protecting children participating in the trial by ensuring that every case was detected and managed quickly. However, this required intense effort

from the study teams, and resulted in identification of a large proportion of transient cases, illustrating the difficulties in diagnosing cases that could have resulted in a need for intervention in routine practice versus incident cases of any severity. This suggests that criteria employed in the trial are inefficient for any form of routine surveillance for intussusception, and future trials may rely MG-132 in vitro on the passive surveillance employed for previous large safety studies. The incidence rate of ultrasound-diagnosed intussusception of 140/100,000 child-years

in the placebo arm is higher than most observational studies but consistent with recent data from Vietnam [18] and is likely attributable to the low threshold for ultrasound evaluation of a potential second case. In the 116E study, the earliest intussusception event in a vaccinated child was 112 days after the third dose. The lack of temporal association between vaccination and event among those vaccinated suggests a causal relationship is very unlikely for cases identified in this trial, but does not preclude a risk similar to that seen with available licensed vaccines. Rotavirus vaccines are recommended for global

use by the World Health Organization [19] and evidence from both developing and developed countries demonstrates the impact of these vaccines on disease reduction in young children [20], [21], [22] and [23]. Increased risk of intussusception has been detected in Australia, Mexico, Brazil and the USA, but the risks of intussusception outweigh the potential benefits of vaccination in disease and mortality reduction, particularly in areas where diarrheal disease continues to be a major killer of children. Nonetheless, monitoring safety will continue to be critical both pre-licensure and after introduction because vaccination safety at the level of the individual child and of programs is necessary to manage rare side effects and to prevent undue harm from newly developed vaccines.

[14] The NC-1 amino acid sequence corresponding to SKSSITITNKRLT

[14]. The NC-1 amino acid sequence corresponding to SKSSITITNKRLTRK [2] was analysed for sequence similarity to other sequences from Taeniidae specimens using the Basic Local Alignment Search Tool (BLAST) algorithm [17] on the National Center for Biotechnology Information public database ( In June 2011, each search was limited to just a single organism whose alignment had an E-value lower than 1.0. The following Taeniidae non-redundant (nr) sequence databases were accessed: T. crassiceps, T. solium, Taenia saginata, Taenia hydatigena,

Taenia multiceps, Taenia pisiformis and Taenia taeniaeformis. The theoretical isoelectric point (pI) and molecular weight (Mw) of Taenia sp proteins were obtained from the Compute pI/Mw Program [18] at Expasy ( In the first immunisation, mice were injected subcutaneously into the intra-scapular fold with one dose, i.e. Antidiabetic Compound Library research buy 20 μg of NC-1 peptide coupled to BSA (NC-1/BSA), TcCa, or BSA dissolved in 50 mM phosphate buffered saline, pH 7.4 (PBS) and emulsified with complete Freund’s adjuvant (1:1, this website volume ratio) in a total volume of 100 μL. Following the guidelines of the animal ethics committee, the boost immunisation using the same route was avoided due to lesions caused by the complete Freund’s adjuvant, and at 2-week intervals, animals received

new intra-peritoneal doses of immunogens emulsified with incomplete

Freund’s adjuvant. One week after the fourth and eighth immunisation, approximately 50 μL of blood was collected from the mice by retro-orbital bleeding to measure antibody reactivity with ELISA. Plates with 96 wells (Falcon Labware, Oxnard, CA) were coated during 16 h at 4 °C with 10 μg/mL of the 3 antigens (non-coupled NC-1 peptide, TcCa, and BSA) dissolved in 50 mM carbonate buffer pH 9.6. After blocking with 2% (w/v) casein diluted in 50 mM Cediranib (AZD2171) phosphate buffered saline, pH 7.4 (PBS) and 0.05% (v/v) Tween 20, the mouse sera against each antigen diluted 1:100 in incubation buffer (Tween 20, 0.25% (w/v) casein) was added to each well and incubated at 37 °C for 1 h. The binding antibody was quantified using goat anti-mouse IgG (whole molecule)-horseradish peroxidase (Sigma # A4416) diluted 1:4000. The reaction was revealed using orthophenylenediamine and H2O2 and stopped by adding 20 μL of 2 N sulfuric acid. Absorbance readings (A492 nm) were carried out in ELISA reader. Following the protocol described above, mice were given a booster 1 week after the second blood sample was obtained. One week later, animals were infected with an intra-peritoneal injection of 5 cysticerci of T. crassiceps resuspended in 100 μL of PBS. Four weeks after this challenge, the animals were euthanised, and peritoneal washing in phosphate-buffered saline (150 mM NaCl, 10 mM sodium phosphate buffer and pH 7.