After 8 h at 40 °C, MVeGFP formulated in formulations C and H suffered <1.0 log loss while the commercial measles vaccines, Attenuvax® and M-VAC™, decreased KRX-0401 supplier by 1.4 logs (1.35–1.53) and 1.9 logs (1.67–2.19), respectively. Assessment

of the formulations by the traditional plaque assay closely correlated with the results of the MVeGFP accelerated degradation assay (Fig. 4b). Overall, the rank order of formulation stability is identical for both methods, supporting the validity of the HT screening strategy. MVeGFP was used as a surrogate for the HT screens because fluorescence is an easily quantifiable endpoint. The most promising formulations were validated using the same non-recombinant measles strains used in commercial vaccines, Edmonston-Zagreb (EZ, used in M-VAC™ from Serum Institute of India) and Moraten (used in Attenuvax® from Merck). Attenuvax and formulated Moraten were thermally challenged at 40 °C for up to 8 h, and infection was quantified following Cellomics data acquisition using the existing MVeGFP algorithm via

an immunofluorescence assay utilizing a FITC-conjugated anti-measles antibody (Fig. 4c). Attenuvax loses 1.0 log (90% counts) of activity after 8 h while formulations A and C only experience Birinapant solubility dmso a ∼0.6 log loss. The tricine-based formulation H exhibited the greatest thermostability, losing only 0.35 log, similar to the results seen with MVeGFP. Interestingly, MVeGFP appears to be less thermally stable than Moraten in the other common formulations. Finally, the most promising formulations were combined with EZ vaccine strain virus, challenged at 40 °C for 4 h, and titered using a plaque assay (Fig. 4d). Non-challenged, formulated virus was

used as a control to calculate log loss and the plaque assay data again supports the HT screening data. The lead candidate formulations are highly stabilizing with no significant loss in activity, whereas the commercial M-VAC™ vaccine suffers >1 log loss. These infectivity data suggest that the two vaccine strains, Moraten and EZ, have differential inherent thermal stability (e.g. formulation C in Fig. 4c vs. d) as has been suggested previously [37] and [38] which Terminal deoxynucleotidyl transferase may result in slightly different behaviors in the same formulation. It is also important to note that while vaccine-strain virus has been used to validate candidate formulations, manufacturing conditions for the commercial vaccines may affect viral stability. For example, it has been reported that the level of cytopathic effect during viral harvest can affect the thermal stability of virus [37]. As proof of concept of broad transferability of the formulation stability screening platform to non-related viruses, the screening process was applied to adenovirus expressing eGFP (Ad-eGFP). A linear response to increasing viral titer was seen with RSDs of 10–20% (Fig. 4e) showing that the assay has similar performance characteristics using either measles or adenovirus.

In addition, the strategy of control programmes based on screening, treatment and contact tracing is extremely costly and requires substantial societal infrastructure. This makes this approach impractical for the developing world, where the burden of disease is the greatest. Thus, development of a safe and effective vaccine is the ultimate goal in the control of Chlamydia. The relative uptake of a vaccine versus screening is difficult to quantify at present, but it is likely that a vaccine would be more widely accepted as evidenced by uptake of the HPV vaccine in settings where it is available and supported [33] and [34]. Costing of a Chlamydia vaccine is not possible at this stage.

However, based on experience from other vaccines, prices could be negotiated to levels that are cost-effective. The most important issue of all is whether Rapamycin research buy a vaccine actually works, that is, has high efficacy and prevents acquisition of infection, transmitting infection or developing disease. This can only be ascertained through clinical research after the development of suitable vaccine candidate(s). With no other long-term strategy available, investment in Chlamydia vaccine design, development and evaluation is the most appropriate way forward. Our objectives in this review are to discuss infections

and diseases LBH589 of the genital tract caused by C. trachomatis with a focus on the complexities and challenges of chlamydial vaccine development. These include considerations such as how to; (i) better understand the range of immunological responses elicited by/to this organism, and therefore to subsequently define effective vaccine antigens and suitable biomarkers of protection, (ii) interpret the results

obtained from animal models of infection, (iii) optimally choose, combine, and present vaccine antigens (surface and/or internal antigens, mucosal adjuvants) and, (iv) interpret mathematical models to define effective vaccine goals for preventing acquisition of infection, interrupting transmission, and/or preventing tubal disease. C. trachomatis is a small (0.5 μm) bacterium that elicits inflammatory cytokine responses following infections of epithelial cells and macrophages. The complex, two-stage developmental cycle of Chlamydia is described Adenylyl cyclase in Fig. 1(a). The extracellular infectious elementary bodies (EB) avoid lysosomal fusion to survive and differentiate into metabolically active reticulate bodies (RB) [35] and [36] and reviewed in [37]). The chlamydial RBs then replicate by around 500-fold, and subsequently re-differentiate into EBs inside a membrane-bound parasitophorous vacuole (“inclusion”) eventually being released by extrusion and/or cytolysis after 40–72 h to infect new cells or hosts [38]. Chlamydia can also enter a persistent growth state if exposed to molecular and cellular stresses such as inadequate antibiotic treatment or host cytokines, particularly IFN-g.

graphpad.com). The data were not normally distributed and hence statistical significance was tested using the Kruskal–Wallis test. When the results were significant, differences among the individual medians were examined using the Mann–Whitney test. Significant effects were declared when P < 0.05. The incorporation efficiency of PTd in the MPs was estimated to be around 78% for PTd and 95%

for CpG and HDP. Previous studies showed that particles less than 10 μm are preferentially taken up by APC [12], [15] and [16]. As such, SEM of MPs that comprised of PCEP with CpG ODN, and IDR-1002 was performed to ensure that the resulting size of the particles was compatible with uptake into APC to ensure that an effective dosage of antigen would be processed. Our previous studies of encapsulated CpG ODN using the same methodology selleck compound not only showed that the MPs generated were less than 10 μm, but also revealed 99% uptake into murine macrophages [12] and [15]. Indeed, the addition of IDR-1002 into the MP was consistent with these previous findings revealing particles ranging in size from 0.5 to 5 μm in diameter (Fig. 1A and B). At higher magnification (20,000×), a close inspection of the surface of these MP revealed that it was not smooth; instead, the surface of these MP seem to be composed

of smaller nanoparticle structures (Fig. 1C). To assess the efficacy of MP formulation, we compared the levels of the pro-inflammatory cytokines selleck chemicals llc TNFα, IL-6 and IL-12p40 in murine J774 macrophages treated with CpG ODN-IDR (AQ), PCEP-CpG ODN-IDR (SOL) and MP co-encapsulating PCEP-CpG ODN-IDR. Other than measuring pro-inflammatory responses, we also looked for the chemokine MCP-1, a chemotactic agent for monocytes/macrophages, T cells, NK cells, and neutrophils, since

it was previously shown that both CpG ODN and the IDR-HH2 alone enhanced MCP-1 mafosfamide production [17], while their complexes demonstrated a synergistic increase in production [11]. The induction of MCP-1 was strongest with the SOL formulation compared to the MP formulation (Fig. 2A) co-encapsulating CpG ODN-IDR complexes or CpG ODN and HDP delivered in uncomplexed MP. The release of pro-inflammatory cytokines TNF-α and IL-6 was significantly higher in MP treated macrophages than AQ or SOL formulation treated groups (Fig. 2B and D). The IL-12p40 levels were two-fold higher in the MP than SOL or AQ formulation treated groups (Fig. 2C). LPS was used as a positive control to demonstrate the viability of the cells. Based on these results, we conclude that the MP delivery induced higher levels of pro-inflammatory cytokines in mouse macrophages.

It seems surprising that physicians thought parents would most likely forego pneumococcal vaccination if MenB vaccination were introduced, since this is a disease at least as severe as MenB IMD with a higher pre-vaccination incidence [17], but maybe less in the focus of privately practicing than of hospital-based pediatricians [18]. However, the other three vaccines named in this context either protect against diseases that are perceived as less severe (rotavirus, varicella) or with a lower risk of infection than MenB IMD (MenC). Age, sex, region and years spent in pediatric practice had a Pictilisib purchase significant effect on some of the responses (Table 2). As age of pediatrician and years

in practice were highly correlated (Pearson’s correlation coefficient = 0.83, p < 0.005), we present results only for the latter. Female physicians and physicians in practice ≥10 years were less likely to fear refusal of other recommended

vaccines if MenB vaccination were introduced, but were more likely to object to simultaneous administration of three vaccines or concomitant MenB vaccination and other vaccines. Correspondingly, female physicians were less likely to prefer Option 1 than their male colleagues, especially females in practice >10 years (see Appendix). Compared to pediatricians from Northern states, pediatricians from Western and Eastern states were more likely and pediatricians from Southern states less likely to believe that parents would be acceptant of MenB vaccination. Southern pediatricians were also more likely to fear refusal of other vaccines if MenB vaccination were recommended, CB-839 particularly if in practice <10 years, while those in Eastern states were less likely enough to fear this, particularly those in practice ≥20 years (Appendix). This corresponds with a lower uptake of standard vaccines in Southern states than in other parts of Germany [19], possibly explained by a higher percentage of anthroposophists/vaccine-sceptics in their population [20] and [21] and a less positive physicians’ attitude towards

vaccination [14]. In contrast, uptake of standard vaccines is highest in Eastern Germany [19], where pediatricians, particularly female pediatricians (Appendix), were most likely to recommend MenB vaccination. Nonetheless, Eastern pediatricians were also more likely to object to simultaneous administration of 3 vaccines and prefer Option 2. Regional differences among German physicians were also seen in a previous study regarding attitude towards pertussis and measles vaccination [14]. As physicians play a crucial role in the implementation and acceptance of new vaccines, assessment of their views is essential. So far, results from only one other study, conducted in 2012 in France, are available on the attitude of pediatricians and general practitioners towards MenB vaccination [22].

S. Centers for Disease Control and Prevention for helpful comments on the manuscript; Ana Rita de Cássia L. Vasconcelos and, most of all, the immunization program team of the Municipal Health Department of Salvador, Brazil. This study was supported by grants from the Bahia State Foundation for the support of research (PP-SUS0001/2009) and National Program of Post-doctoral (CAPES-PNPD 1472/2008). “
“Human papillomavirus (HPV) vaccines have the potential to significantly reduce the incidence of cervical cancer, the leading cause of cancer mortality among women in sub-Saharan Africa [1] and [2]. Two HPV vaccines have

BIBW2992 manufacturer now been approved for use in many countries. These provide a high degree of protection against HPV 16/18 infections and associated cervical lesions [3], [4] and [5]. The World Health Organisation recommends offering HPV vaccine to girls at ages 9–14, prior to sexual debut, since the vaccine has highest efficacy if girls have not already acquired

HPV [6]. Many high-income countries and some middle-income countries have started national HPV vaccination programs, either school-based or on-demand programs, with vaccine coverage (completion of the 3-dose regimen) ranging from 9% (Greece) to 32% (US) and 76% (UK) [7], [8], [9] and [10]. EX 527 mouse In sub-Saharan Africa, two vaccine demonstration projects have been completed [11] and [12]; Rwanda has embarked on a national HPV vaccination programme [13] and [14], and Tanzania plans to start a similar programme in 2012. Research in Africa on HPV vaccine acceptability and delivery is needed to understand how best to deliver this vaccine to adolescent girls among populations who have little or no knowledge about cervical Etomidate cancer, and may be suspicious of vaccines that target young women or a sexually transmitted infection (STI) [15], [16], [17], [18], [19], [20], [21] and [22]. Between August 2010 and June 2011, in preparation for a national HPV immunisation program, a phase IV cluster-randomised trial (NCT01173900) in schoolgirls in Mwanza Region, Tanzania, was conducted to measure the

feasibility, uptake, and acceptability of two school-based HPV vaccine delivery strategies: age-based (all girls born in 1998) or class-based (all girls in Year 6 of primary school in 2010) [12]. We present findings from a qualitative sub-study conducted before the actual HPV vaccination started in August 2010. The sub-study’s objectives were to learn what people knew about cervical cancer and HPV vaccination, whether they would find HPV vaccination acceptable, and how they viewed vaccine delivery and consent procedures. These findings were used to improve sensitisation and vaccination procedures within the trial and to assist preparations for a national HPV vaccination program. The qualitative sub-study study took place in the two districts of Mwanza city and a neighbouring rural district (Misungwi), between March and August 2010.

Kobashigawa Over the last 4 decades, cardiac transplantation has become the preferred therapy for select patients with end-stage heart disease. check details Heart transplantation is indicated in patients with heart failure despite optimal medical and device therapy, manifesting as intractable angina, refractory heart failure, or intractable ventricular

arrhythmias. This article provides an overview of heart transplantation in the current era, focusing on the evaluation process for heart transplantation, the physiology of the transplanted heart, immunosuppressive regimens, and early and long-term complications. David A. Baran and Abhishek Jaiswal From humble beginnings in 1963 with a single desperately ill patient, mechanical circulatory support has expanded exponentially to where it is a viable alternative for advanced heart failure patients. Some of these patients are awaiting transplant but others will have a mechanical heart pump as their ultimate

treatment. The history of MCS devices is reviewed, along with the 4 trials that define the modern era of circulatory support. The practical aspects of life with an MCS device are reviewed and common problems encountered with MCS devices. Future trends including miniaturization and development of completely contained MCS systems are reviewed. Heath E. Saltzman Atrial fibrillation and ventricular tachyarrhythmias are frequently seen in patients with heart failure, and complicate the management of such patients. Talazoparib price Both types of arrhythmia lead to increased morbidity and mortality, and often prove to be challenging issues to manage. The many randomized studies that have been performed in patients with these conditions and concomitant heart failure new have helped in designing optimal treatment strategies. Liviu Klein and Henry Hsia Sudden cardiac deaths account for 350,000 to 380,000 deaths in the United States annually. Implantable cardioverter-defibrillators have improved sudden death outcomes in patients with heart failure, but only a minority of patients with defibrillators receives appropriate therapy for ventricular arrhythmias. The risk prediction for sudden death and selection of patients

for defibrillators is based largely on left ventricular ejection fraction and heart failure symptoms because there are no other risk stratification tools that can determine the individual patients who will derive the greatest benefit. There are several other pharmacologic strategies designed to prevent sudden death in patients with heart failure. Daniel F. Pauly Acute decompensated heart failure may occur de novo, but it most often occurs as an exacerbation of underlying chronic heart failure. Hospitalization for heart failure is usually a harbinger of a chronic disease that will require long-term, ongoing medical management. Leaders in the field generally agree that repeated inpatient admissions for treatment reflect a failure of the health care delivery system to manage the disease optimally.

14%; mp 214 °C; IR (KBr) vmax 2967, 1540, 1390, 1170, 1180, 756 cm−1; 1H NMR (CDCl3) δ ppm; 7.32–8.10 (m, 11H, Ar–H), 2.99 (s, 3H, SCH3); 13C NMR (CDCl3) δ ppm; 158.2, 148.2, 144.2, 141.3, 1139.2, 138.3, 134.2, 133.4, 130.2, 130.0, 129.9, 129.2, 128.3, 128.0, 127.5, 127.1, 125.1, 123.4, 15.3; HRMS (EI) m/z calcd for C22H13 Cl N3 O2 S2: 451.0216; selleck kinase inhibitor found: 451.0212. This compound was prepared as per the above mentioned procedure purified and isolated as yellowish solid: yield 91.3% mp 207 °C; IR (KBr) vmax 2956,1545, 1417, 1320, cm−1; 1H NMR (CDCl3) δ ppm; 7.08–8.01 (m, 11H, Ar–H), 3.87 (s, 6H, OCH3); 13C NMR (CDCl3) δ ppm; 162.3, 158.2,

149.3, 144.2, 139.2, 138.6, 132.6, 131.6, 128.6, 127.4, 125.2, 125.0, 123.7, 115.3, 56.3; HRMS (EI) m/z calcd for C23H17N3O4S: 431.4638; EPZ5676 ic50 found: 431.4634. The compound was prepared

as per the general procedure mentioned above purified and isolated as yellow solid; yield 88.23%; mp 203 °C; IR (KBr) vmax 2920, 1534, 1320, 1170, 712, cm−1; 1H NMR (CDCl3) δ ppm; 7.40–7.68 (m, 10H, Ar–H), 2.22 (s, 3H, CH3); 13C NMR (CDCl3) δ ppm; 158.2, 149.3, 145.6, 140.2, 139.5, 138.6, 137.5, 134.6, 130.3, 130.1, 129.4, 129.1, 127.3, 127.0, 126.3, 126.0, 123.4; HRMS (EI) m/z calcd for C22H13Cl2N3O2S: 453.0106; found: 453.0102. The compound was prepared as per the general procedure mentioned above purified and isolated as colorless solid; yield 73.02%; mp 214 °C; IR (KBr) vmax 2954, 1545, 1390, 1270, 757 cm−1; 1H NMR (CDCl3) δ ppm; 7.34–8.10 (m, 10H, Ar–H), 2.54 (s, 3H, SCH3); 13C NMR (CDCl3) δ ppm; 157.3, 149.7, 145.8, 142.4, 139.8, 138.7, 137.5, 135.7, 132.4, 132.4, 131.4, 131.5, 130.4, 129.4, 129.1, 128.7, 127.4, 127.2, 127.0, 126.8, 124.5, 121.4; HRMS (EI) m/z calcd for C22H14Cl2N3O2S2: 484.9826; next found: 484.9821. This compound was prepared as per the above mentioned procedure purified and isolated as yellowish solid: yield 53.05% mp 198 °C; IR (KBr)

vmax 2974, 1477, 1275, 570 cm−1; 1H NMR (CDCl3) δ ppm; 7.16–8.0 (m, 11H, Ar–H), 3.94 (s, 6H, OCH3); 13C NMR (CDCl3) δ ppm; 162.3, 157.8, 139.8, 139.0, 138.2, 134.6, 131.6, 130.4, 128.9, 125.6, 124.7, 123.8, 117.8, 115.7, 56.3; HRMS (EI) m/z calcd for C23H17BrN2O2S: 464.0194; found: 464.0190. This compound was prepared as per the above mentioned procedure purified and isolated as slight yellowish solid: yield 66.89% mp 186 °C; IR (KBr) vmax 29782, 1320, 1120, 650, cm−1; 1H NMR (CDCl3) δ ppm; 7.38–8.10 (m, 11H, Ar–H), 3.86 (s, 3H, OCH3); 2.98 (s, 3H, SCH3); 13C NMR (CDCl3) δ ppm; 162.7, 158.3, 141.4, 139.8, 139.0, 138.4, 132.4, 131.5, 131.0, 128.4, 128.0, 127.6, 127.2, 124.3, 123.7, 116.3, 115.6, 56.2, 15.6; HRMS (EI) m/z calcd for C23H17BrN2OS2: 479.9966; found: 479.9961.

We collected information on personal characteristics (age, gender), mumps-related symptoms (using visual prompts), complications, possible previous mumps infections, contact with mumps cases, days absent from social activities, contact with health care providers and self-reported immunization status. We used a web-based questionnaire (Lime survey software, version 1.91). We sent MK-2206 datasheet invitations to the selected students on the 18th of March 2013, followed by a reminder one week later. We reviewed the medical files of the university medical service to obtain the documented immunization status of participants. We described mumps cases by time, place

and person. We calculated relative risks (RR) of mumps according to immunization status and a selection of risk factors along with 95% confidence intervals. We considered a p-value <0.05 as statistically significant. We extrapolated the incidence of self-reported parotitis to the complete student population of the KU Leuven. We calculated vaccine effectiveness (VE) as the difference in attack rate between those vaccinated twice and those vaccinated once over the attack rate in those

vaccinated once. We calculated the time in years since the second vaccination based on the documented vaccination data. We analyzed data using STATA 12.00 (STATA Corporation, College Station, TX, USA) and SAS 9.3 (SAS Institute Inc. 2011, Raf inhibitor TX, USA). Informed consent from all students who were included in the study was obtained. On December 14, 2012, the ethics committee of the hospital of KU Leuven approved the study protocol. Between June 16, 2012 and April 16, 2013, 4052 cases were reported from Flanders, of which 1187 were possible, 1294 were probable and 1540 were laboratory-confirmed (overall reported rates: 31.5/100,000 population). Calpain Reported cases of mumps peaked in December 2012 (Fig. 1). Most cases were reported in cities where universities are located, including

Ghent (n = 510), Leuven (n = 419), Kortrijk (n = 415) and Antwerp (n = 365) ( Fig. 2). Fifty-eight percent (n = 2364) of the cases were male and 58% (n = 2348) were between 15 and 25 years of age. Vaccination information was available for 1190 (29%) cases. Of these, 70% (n = 836) were vaccinated twice, 28% (n = 338) were vaccinated once and 2% (n = 16) were unvaccinated. Orchitis was reported in 11% (n = 145) of male cases for whom the status of complications was known. Other complications included meningitis (n = 8; 0.2%) and pancreatitis (n = 5; 0.1%). Between June 16, 2012 and April 16, 2013, 128 specimens were collected from Flanders and tested for mumps virus at the NRC. All specimens were tested by PCR; 53% were confirmed. Genotyping was performed in41 specimens.

The survey also included an open text field for feedback. We identified our survey sample from the VHA Cardiac Assessment Reporting and Tracking — Catheterization Laboratory (CART-CL) system, a national, real-time database used in all VHA cardiac catheterization

laboratories to record cases [10]. Our sampling frame was all VHA interventional cardiologists registered in the CART-CL system as of December 13th, 2012 and we drew a 100% sample. The survey was fielded in February 2013 using Inquisite software (Allegiance Inc., Austin, TX), a Web-based survey tool. The survey link was e-mailed to participants up to 10 times over a 5 week period. Surveys were anonymous. We linked surveys to site-level data RO4929097 solubility dmso on the number of total PCIs and number of TRIs performed from CART, in order to report perceptions of the relative superiority

of TRI and barriers to TRI stratified by cath-lab TRI rates. We did not conduct statistical comparisons due to insufficient sample size. Radial proportion check details was the site-level number of TRI cases divided by TRI plus TFI cases for the 2013 calendar year. This study was reviewed and approved by the Central Institutional Review Board for the Department of Veterans Affairs, Research and Development Office, with a Waiver of Documentation of Informed Consent for the cath lab staff participating in the training and for the survey respondents (VHA Central IRB #12-10). Copies of the interview guide and survey are available from the authors upon

request. We received 78 completed surveys (32% response rate) from 48 of the 65 cath labs where interventional cardiologists were surveyed (survey data received from Calpain 73% of sites). The majority of respondents (Table 1) had been practicing for 6 or more years and reported using radial access for fewer than 25% of diagnostic or interventional cases. A plurality of respondents (41%) reported that 80% or more of their PCI cases were performed immediately after diagnostic angiography was completed (i.e., ad hoc) as opposed to scheduling the patient for a separate PCI at a later date (scheduled). In general, attitudes favored radial access (Table 2) with respondents rating radial access “somewhat better,” “better” or “much better” in terms of ease of monitoring patients following the procedure (70.8%), allowing patients to go home sooner (76.9%), fewer vascular access complications (83.1%), comfort for patients (84.6%), and fewer bleeding complications (93.8%). Conversely, overall, a minority of respondents rated radial access somewhat better, better or much better in terms of how fast they could complete the procedure (9.

Written and signed informed

consent was obtained from parents or guardians of participating children for vaccination and sampling procedures. PCV7 was provided by Wyeth Lederle Portugal (Farma), Lda. The vaccinated group was immunized with a single dose of the vaccine in May 2001. The intramuscular injection of 0.5 mL of vaccine was performed by a pediatric nurse in the deltoid muscle of the upper arm of each child. Pediatric nurses collected the nasopharyngeal specimens by use of calcium alginate swabs (BBL Culture Swab; Becton-Dickinson, Sparks, MD). Swabs were inserted through the child’s nostril until they touched the posterior nasopharynx, rotated 180°, removed, placed in transport media Galunisertib (Stuart medium) and transported at room temperature to the Laboratory of Molecular Genetics at Instituto de Tecnologia Química e Biológica.

Bacterial samples were processed within 4 h of collection [25]. Each child from the vaccinated and control groups was sampled in May and June 2001. In the vaccinated group, the first nasopharyngeal sample was collected immediately before immunization with a single PCV7 dose, in May 2001. Children carrying pneumococcal isolates expressing only check details one capsular type (serotype) were designated as single carriers and children carrying more than one serotype were designated as multiple carriers. Among the latter, the serotype found in the majority of the isolates (>50%) was designated as the dominant serotype and the remaining serotypes were named minor serotypes. The ecological mechanisms that could be identified in this study were defined as follows: (i) clearance (disappearance of a pneumococcal isolate of a given serotype); (ii) de novo acquisition (acquisition of a new pneumococcal isolate of a given serotype); (iii) unmasking (expansion of a minor serotype that becomes the dominant serotype); (iv) maintenance (maintenance of a given serotype) and (v) capsular switch (an isolate maintains its genotype/PFGE pattern, but

presents a different serotype). Each nasopharyngeal swab was ADP ribosylation factor streaked onto 5 μg/mL gentamicin-5% sheep blood triptic soy agar plate and incubated at 37 °C in 5% CO2 atmosphere. Whenever available, up to 10 pneumococcal colonies were picked from this primary plate. Colonies were chosen randomly and any morphologically distinct colony was also picked. Colonies were re-streaked and cultivated on 5% sheep blood triptic soy agar and frozen at −80 °C in Mueller-Hinton broth containing 15% glycerol (v/v). Phenotypic characteristics (optochin susceptibility, morphology, and α-hemolysis) were used for presumptive pneumococci identification. The bile solubility assay was performed on suspected pneumococcal cultures exhibiting decreased susceptibility to optochin. These purified cultures were used in the subsequent assays. All pneumococcal isolates were serotyped by the Quellung reaction using specific capsular antisera (Statens Seruminstitut, Copenhagen, Denmark) [26].