Search

Background: Migraine is a complex multifactorial disorder and its pathogenesis still remains unclear. Evidence suggests the involvement of the activated trigeminovascular pathway, in which BDNF seems to play an important role. Therefore, BDNF polymorphisms are promising candidate susceptibility factors.Aim: BDNF rs6265 functional polymorphism was analyzed in order to determine its possible association with pediatric headache and migraine risk.Methods: The research included 120 consecutive pediatric patients who were diagnosed with headache and 120 healthy controls. The diagnosis was in compliance with the International Classification of Headache Disorders. Blood samples were collected from all participants and genotyped for rs6265.Results: BDNF rs6265 was significantly associated with decreased headache risk, particularly in the dominant model [Odds Ratio, OR (95% confidence interval, C.I.): 0.47 (0.26-0.85), p = 0.011] and the log-additive model [OR (95% C.I.): 0.48 (0.28-0.82), p = 0.0053]. During the sensitivity analysis, the associations were also maintained among patients with migraine.Conclusions: This is the first study to reveal a significant association of this BDNF variant with headache risk. Additionally, Val66Met was also for the first time related to decreased childhood migraine risk.

Purpose: To identify changes in the gait kinematics and temporospatial parameters of children with bilateral Cerebral Palsy (CP) at 8 months after the onset of independent walking and identify differences to Typical Development (TD) children at the onset of independent walking and at 8 months follow up.Method: Sixteen children with bilateral CP, GMFCS levels I and II, and 15 TD children were recruited. Gait kinematics and temporospatial parameters were recorded using a 3-D gait analysis system; the sagittal plane of the lower limb joints was analyzed. Baseline measurements were recorded at the individual's onset of independent walking and follow up was after 8 months.Results: Compared to baseline, children with bilateral CP demonstrated increased (mean difference ± SE) plantar flexion (11.79 ± 2.96), single support (0.04 ± 0.01), step length (0.2 ± 0.05) and stride length (0.4 ± 0.09), at follow up; all p < .05. Compared to TD children, they also had lower gait speed (0.16 ± 0.05), higher single support (0.02 ± 0.01) and lower maximum knee extension (9.14 ± 4.49) during the swing phase, at baseline and follow up (0.1 ± 0.04, 0.05 ± 0.01, 23.04 ± 4.17, respectively); all p < .05.Conclusion: There are changes in the sagittal plane kinematics and temporospatial parameters of the gait during the first 8 months of independent walking. These indicate gait maturation changes and highlight the impact of walking experience on the gait characteristics of children with bilateral CP.

Breastfeeding rates remain extremely low in Greece and women with gestational diabetes mellitus and hypothyroidism may experience additional difficulties. The aim of the study was to investigate the effect of a structured individualized lactation educational intervention by a midwife on increasing breastfeeding rates in women with endocrine disorders and low-risk women compared to women receiving standard care, 24 months after delivery. Two-hundred women made up the study population. Half of them were experiencing endocrine pregnancy disorders and 100 women constituted the low-risk pregnancy standard care control group. Women who were breastfeeding exclusively were significantly higher in the midwifery intervention group with endocrine disorders, namely breastfeeding continued at four months (breastfeeding: 20% vs. 12%, exclusive breastfeeding: 50% vs. 26%, p = 0.0228), and at six months after childbirth (breastfeeding: 54% vs. 28%, exclusive breastfeeding: 32% vs. 12%, p = 0.0011), compared to the standard care control group with endocrine disorder. The low-risk midwifery intervention group breastfed at four months (22% vs. 14%, p = 0.0428) and at six months (52% vs. 26%, p = 0.0018) at higher rates compared to the standard care control group. In addition, exclusive breastfeeding was significantly higher in the low-risk midwifery intervention group at four months (46% vs. 20%, p = 0.0102) and six months (38% vs. 4%, p &lt, 0.0001) compared to the standard care control group. This study was the first attempt of a structured midwifery breastfeeding education in Greece and its major contribution reflects a significant positive impact on breastfeeding rates in terms of duration and exclusivity in women with gestational endocrine disorders as well as in low-risk women, and could possibly be applied and instituted in everyday clinical practice to increase the low breastfeeding rates in Greece.

Purpose: To determine the efficacy of scleral buckling in eyes with stage 4A and 4B retinopathy of prematurity (ROP).Methods: Seven eyes of five premature infants underwent scleral buckling for stage 4 ROP in zone II. Five eyes had stage 4A ROP, and two eyes had stage 4B ROP. Six eyes had previous diode laser photocoagulation, and one eye had received an intravitreal ranibizumab injection. Scleral buckling was the procedure of choice due to lack of access to specialized pediatric vitrectomy instrumentation. Average age at surgery was 3.4 months. Postoperative anatomic retinal status, visual acuity outcome and refractive error were assessed. Results: The scleral buckle was removed on average 8 months after surgery. Retinal reattachment was achieved in all seven eyes. At final follow-up one eye had macular ectopia and disc dragging, one eye had a macular traction fold and two eyes had optic disc pallor. Average myopic error after buckle removal was -7.5 D.Conclusion: Scleral buckling can be performed safely and effectively in 4A and 4B stage ROP in critically-ill infants, when access to specialized pediatric vitrectomy instrumentation is limited. This surgical technique may provide adequate relief of vitreoretinal traction with improved visual potential.

Backround The impact of the consistent implementation of “aggressive” nutrition by means of intensive early neonatal nutritional support up to 40–44 weeks postmenstrual age and the use of nasal continuous positive airway pressure (nCPAP) within the first hour of life on the respiratory function of very premature neonates (VPN) at school age is unclear. Method Respiratory function was evaluated in 108 VPN and 70 term controls. Growth, frequency of lower respiratory tract infections, re-hospitalization, and spirometry were recorded up to 8–10 years of age. Comparison was carried out between the two study groups. Results There was no significant difference in forced expiratory volume in 1 s and forced vital capacity at 8 years of age, and also in lower respiratory tract infections and re-hospitalization due to them, up to 8 years of age between preterm and term neonates. No significant difference was found in spirometry measurements neither between premature neonates with and without BPD nor between the two subgroups of preterms and term neonates. Conclusion “Aggressive” nutrition, persistent nCPAP use, and their impact on early postnatal growth probably positively affect the respiratory function of our study population. These very encouraging results need to be confirmed by larger studies.

Introduction: The effects of “aggressive” neonatal feeding policies of very preterm neonates (VPN) and the risk of metabolic syndrome later in life remain questionable. We aimed to evaluate the effect of our “aggressive” nutrition policies of VPN during hospitalisation on body mass index (BMI) at ages 2 and 8 years. Materials and Methods: Eighty four VPN, who received “aggressive” nutrition during hospitalisation in an effort to minimise postnatal growth restriction (PGR) (group A), and 62 term neonates, as controls (group B), were enrolled in the study. Group A was further divided in four subgroups depending on the type (A1: fortified expressed breast milk and preterm formula, A2: exclusively preterm formula) and quantity of milk received (A3: maximum feeds 180–210 mL/kg/day, A4: maximum feeds 210 and up to 260 mL/kg/day). BMI was calculated at ages 2 and 8 years and plotted on the centile charts. Results: There was no significant difference in BMI between groups A and B at 2 and 8 years, respectively, in both absolute BMI values and their centile chart distribution. There was no significant difference in BMI at 2 and 8 years either between subgroups A1 and A2 or between subgroups A3 and A4. Conclusions: “Aggressive” and individualised feeding policy for VPN did not affect the BMI and obesity rates at ages of 2 and 8 years in our study population. The type and quantity of milk feeds had no impact on their BMI at school age. Further larger studies are needed to confirm our results.

Background: We aimed to evaluate gastric volume changes during intermittent milk feeds (IMF) and continuous milk feeds (CMF) in very premature neonates (VPN), with gastric residual volume (GRV) based on antral cross-sectional area (ACSA) measurements and to examine if there were differences in GRV between the two feeding methods. Methods: A randomized prospective clinical trial with crossover design was conducted in 31 preterm neonates (gestational age &lt, 30 weeks). Gastric volume was assessed twice in each neonate (during IMF and CMF feeding), at 7 specific time points during a 2-h observation period by measuring ACSA changes via the ultrasound (U/S) method. Results: There was a significantly different pattern of gastric volume changes between the two feeding methods. GRV, expressed as the median percentage of ACSA measurement at 120 min relative to the higher ACSA measurement during IMF, was found to be 3% (range 0–25%) for IMF and 50% (range 15–80%) for CMF. Neonates fed with IMF had a shorter mean gastric emptying time compared to those fed with CMF (p = 0.0032). No signs of feeding intolerance were recorded in either group during the period of observation. Conclusions: Our results showed that gastric volume changes and gastric emptying time in VPN, based on ACSA measurement changes, depend on the milk feeding method. No gastrointestinal complications/adverse events were noted with GRV up to 80% with CMF.

As most children infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) present with mild symptoms or they are asymptomatic, the optimal strategy for molecular testing it is not well defined. The aim of the study was to determine the extent and aetiology of molecular testing for SARS-CoV-2 in Greek paediatric departments during the first phase of the pandemic and identify possible differences in incidence, depending on the age group and geographical area. We conducted a nationwide study of molecular testing for SARS-CoV-2 of children in paediatric departments between March and June 2020. A total of 65 paediatric departments participated in the study, representing 4901 children who were tested for SARS-CoV-2 and 90 (1.8%) were positive. Most paediatric cases were associated with topical outbreaks. Adolescents 11–16 years had the highest positivity rate (3.6%) followed by children 6–10 years (1.9%). However, since the testing rate significantly differed between age groups, the modified incidence of SARS-CoV-2 infection per age group was highest in infants 5 population). Most children tested presented with fever (70.9%), respiratory (50.1%) or gastrointestinal symptoms (28.1%). Significant differences were detected between public and private hospitals regarding the positivity rate (2.34% vs. 0.39%, P-value

Background: Caffeine has been commonly used for prevention and treatment of apnea-related symptoms in premature infants. However, its side effects have not been thoroughly studied. We investigated whether caffeine affects gastric motility in very-preterm (VP) neonates. Methods: The study is a randomized crossover clinical trial. Twenty-two neonates with mean birth weight (BW) (standard deviation&mdash, SD) 1077 (229) g and mean gestational age (GA) (SD) 28.6 (2.1) weeks were recruited. Each neonate had its gastric emptying time checked twice with ultrasound assessment of changes in antral cross sectional area (ACSA). All neonates were sequentially allocated to the caffeine group (A) and the control group (B). Complications from the gastrointestinal tract were documented throughout the study. Results: Statistically significant difference was found with regards to the gastric emptying time [median, (range)] between caffeine and control group (p = 0.040). Additionally, in the neonates with BW 1000&ndash, 1500 g and GA &ge, 28 weeks, the gastric emptying time (minutes) was significantly longer during caffeine treatment [44.5 (36&ndash, 68.2)] and [40 (34.5&ndash, 66.5)] respectively, as compared to the gastric emptying time during no caffeine treatment [27 (24.2&ndash, 30)] (p = 0.002) and [27 (24.5&ndash, 30)] (p = 0.001). The incidence of gastrointestinal (GI) complications was significantly greater in neonates receiving caffeine [6 (27.%)] as compared with those without caffeine treatment [1 (4.6%)] (p = 0.039). Conclusions: During caffeine treatment, a significantly delayed gastric emptying time was noted in all study neonates, especially in these with BW 1000&ndash, 1500 g and those with GA &ge, 28 weeks. Further larger studies are necessary in order to confirm this interesting finding.

Background The surveillance of emm types and macrolide susceptibility of group A streptococcus (GAS) in various areas and time periods enhances the understanding of the epidemiology of GAS infections and may guide treatment strategies and the formulation of type-specific vaccines. Greece has emerged as a country with high macrolide use. However, studies suggest a gradual reduction in macrolide consumption after 2007. Methods During a 7-year period (2011–2017), 604 GAS isolates were recovered from consecutive children presenting with pharyngeal or nonpharyngeal infections in Central Greece; 517 viable isolates underwent molecular analysis, including emm typing. Results Isolates belonged to 20 different emm types (in decreasing order of prevalence: 1, 89, 4, 12, 28, 3, 75 and 6, accounting for 88.2% of total isolates). The emm types comprised 10 emm clusters (five most common clusters: E4, A-C3, E1, A-C4 and A-C5). The emm89 isolates were acapsular (‘new clade‘). Overall macrolide resistance rate was 15.4%, and cMLSB emerged as the predominant resistance phenotype (56.4%). The lowest annual resistance rates occurred in 2014 (13.1%), 2016 (5.5%) and 2017(8.0%) (P for trend = 0.002). Consumption of macrolide/lincosamide/streptogramin B declined by 22.6% during 2011–2017. Macrolide resistance and emm28 and emm77 types were associated (both P

Molecular analysis of different types of thumb duplication and identification of new suspected gene mutations. In a series of patients operated for polydactyly, DNA was extracted from blood samples collected preoperatively. Among these, the samples of two patients with thumb duplication (Wassel types III and IV) were initially selected for molecular analysis. The method of Clinical Exome Solution was used for the study of the phenotype-involved genes. Next-generation sequencing (NGS) was performed on a NextSeq-500 Platform (Illumina), and Sophia DDM® SaaS algorithms were used for the bioinformatics analysis of the data. In total, 8—including 4 new—mutations were detected in CEP290 (1 mutation), RPGRIP1 (2 mutations), TMEM216 (2 mutations), FBN1 (1 mutation), CEP164 (1 mutation), and MEGF8 (1 mutation) genes. NGS revealed 3 mutated genes in the patient with Wassel III thumb duplication and 5 mutated genes in the patient with Wassel IV duplication. The molecular analysis revealed that the patients had 2 mutated genes in common, but they only shared one common mutation. The new detected mutations are most probably associated with thumb duplication, as they belong to genes with already described mutations causing ciliopathies, often including polydactyly in their phenotype. Recognition of these mutations will be helpful to prenatal diagnosis, operative treatment strategy prediction, and possible future experimental applications in gene therapy.

Purpose The purpose of this study was to prospectively investigate the association between retinopathy of prematurity (ROP) and ocular growth in premature infants during the earliest weeks of life. Methods Premature infants in the national ROP screening program were recruited and examined at 1- or 2-week intervals between 30 and 38 weeks of postmenstrual age. One hundred infants with gestational age (GA) between 24 and 35 weeks (30.04 + 2.13), and birth weight (BW) between 550 and 2060 g (1251.45 + 317.19) were included in the study. At each examination, the presence, stage, and zone of ROP were recorded along with axial length (AL), central corneal thickness (CCT), and weight gain. Biometric parameters were measured by A-scan biometry. Study variables included GA, BW, AL, CCT, weight gain, relative weight (RW), and dif_AL, dif_CCT, and dif_weight, which are the differences between two consecutive recordings of the same infant. Multiple regression analysis models were used to determine the association between the study variables and ROP. Results dif_AL, dif_CCT, and RW were the most appropriate variables to detect the optimal threshold points that discriminate ROP: weekly increase of AL < 0.095 mm, weekly reduction of CCT < 0.5 μm, or weekly weight gain < 7% is associated with ROP development. Conclusions ROP is associated with delayed ocular development, as eyes of premature infants with ROP have shorter axial lengths and thicker corneas than eyes of premature infants without ROP. The association of AL, CCT, and weight gain with ROP could be of value for future development of predictive models for ROP.

We have read the article entitled “Applying Methods for Postnatal Growth Assessment in the Clinical Setting: Evaluation in a Longitudinal Cohort of Very Preterm infants” by Montserrat Izquierdo Renau et al [...]

Background We evaluated the long-term impact of full PCV13 vaccination in a 3 + 1 schedule on pneumococcal colonization patterns of children in order to clarify PCV13 serotype persistence/enhancement and re-colonization. Methods From January 18 to August 29, 2017, consecutive children who had received the 4-dose course of PCV13, as per the National Immunization Program recommendations, were prospectively enrolled through 45 general pediatric practice facilities in 30 municipalities in Greece. A single oropharyngeal sample was obtained from each subject in a standardized manner (questionnaire, procedure). Based on the time interval since the fourth dose of PCV13, the children sampled were grouped for analysis in 6 groups: 26 days to 11 months; 12–23 months; 24–35 months; 36–47 months; 48–59 months, and 60–71 months. Carriage and distribution of Streptococcus pneumoniae serotypes was detected by RT–PCR. Results A total of 1212 children aged 14–83 months were investigated. S. pneumoniae was identified in the pharyngeal swab of 617 children (50.9%); 172/617 (27.9%) children carried > 1 pneumococcal serotypes. As a consequence of co-colonization, a total number of 718 S. pneumoniae (belonging to 28 serotypes) was identified. The carriage rate of non-PCV13 serotypes escalated within 3 years after the fourth dose and plateaued during the fourth and fifth year. The carriage rate of PCV13 serotypes escalated during the 4 years after the fourth dose and declined thereafter. 22/305 children (7.2%) carried one or more PCV13 serotypes in the first year after the fourth vaccine dose, 27/201 (13.4%) in the second year, 34/207 (16.4%) in the third year, 48/224 (21.4%) in the fourth year, 40/191 (20.9%) in the fifth year and 13/84 (15.5%) in the sixth year (P < 0.0001) (Figure 1). The colonization frequency of serotypes 3 and 19A increased with the rise of the vaccination time interval (Figure 2). Changes in the frequency of other PCV13 serotypes were not significant. Serotypes 7F, 14 and 23F were not recovered. Conclusion Our study suggests that S. pneumoniae is present in the pharynx of children 26 days to 71 months after the completion of PCV13 vaccination, and that non-PCV13 serotypes predominate throughout this period. The carriage rate of PCV13 serotypes 3 and 19A increases significantly as the time interval from the fourth dose of PCV13 increases. Disclosures All authors: No reported disclosures.

In Greece, there are high rates of methicillin (40%-60%) and clindamycin (15%-25%) resistance among community-acquired Staphylococcus aureus isolates. Therefore, we sought to identify other antimicrobial treatment options such as daptomycin.We studied retrospectively all pediatric infections treated with daptomycin at the University General Hospital of Larissa, Greece, from January 1, 2007, to June 16, 2016.Of a total of 128 patients (median age: 2.8 years; range: 8 days to 14.5 years; 76.6%7 years) treated with daptomycin, 45 (35.2%) had invasive infection, most frequently musculoskeletal, and 83 (64.8%) had noninvasive infection, that is, complicated skin and soft tissue infection. S. aureus was the most commonly recovered pathogen (n = 61) (63.9% methicillin-resistant isolates, 21.3% clindamycin-resistant). The average daily dose of daptomycin was 10 mg/kg qd, and the median duration of therapy was 10 days. Daptomycin was administered alone (n = 61) or in combination therapy (n = 67), most frequently with rifampin (n = 40) and/or a β-lactam antibiotic (n = 33). Open or closed drainage was performed in 86 (67.2%) of the total number of patients. Of 128 treated patients, 123 (96.1%) achieved clinical success, 114 (89.1%) had complete remission, and 9 (7%) had improvement of their disease. There were no failures with daptomycin therapy. The adverse events were of no clinical significance.Daptomycin administered alone or in combination with other antimicrobial agents to children was efficacious and well tolerated in the treatment of complicated infections of suspected or proven staphylococcal etiology.

Parapneumonic effusions in children are usually associated with pneumococcal infections. In Greece, the 7-valent pneumococcal conjugate vaccine was replaced by higher-valent pneumococcal conjugate vaccines (PCVs); 10-valent was introduced in May 2009 and 13-valent (PCV13) in June 2010. Since July 2010, PCV13 has been the most commonly used PCV. In a study conducted at the University General Hospital of Larissa, Central Greece, from January 2012 to January 2016, 85.7% of children born after the implementation of PCV13 and aged 24-59 months had received the complete series (3 + 1 immunization schedule) of PCV13.We studied all paediatric community-acquired pneumonia cases with empyema hospitalized at the University General Hospital of Larissa from January 2008 to January 2016.There were 30 cases of parapneumonic empyema. Among 27 empyema cases of known aetiology, 19 (70.4%) were due to Streptococcus pneumoniae (identifiable serotypes 3, 19A, 7F, and 9N/L). After September 2011, no more cases caused by serotypes 7F and 19A were observed, whereas serotype 3 emerged as the predominant pathogen of pneumococcal empyema (9 of 11 cases). Serotype 3 continued to cause empyema despite vaccination with PCV13 either fully with a 3 + 1 schedule (n = 3) or with one booster dose at the age of 21 months (n = 1).In Central Greece during the first five years of high coverage with PCV13, serotype 3 was the only PCV13 serotype that clearly persisted in children with empyema.

In order to study whether the use of the heptavalent pneumococcal conjugate vaccine (PCV7) led to a shift in the Streptococcus pneumoniae serotypes distribution and whether it modified the resistance to antibiotics, 2649 nasopharyngeal samples were obtained between 2005 and 2009, from children attending day-care centers in Central Greece. The percentage of attendees vaccinated with ≥1 dose of PCV7 increased from 12.9% (2005) to 95.5% (2009). Non-PCV7 serotypes replaced those belonging to PCV7. In 2009, 19F was virtually the only PCV7 serotype that continued to circulate. A significant increase in the frequency of penicillin-intermediate (oral penicillin V breakpoints) isolates coincided with a marked reduction in isolates with high resistance to penicillin. Several non-PCV7 serotypes colonized the children, but their frequency varied substantially from year to year. Each one of 14 specific non-PCV7 serotypes, i.e. 6A, 11A, 15B, 23A, 10A, 16F, 38, 22F, 15C, 19A, 35F, 24F, 6C, and 7F, accounted for ≥2% of pneumococcal isolates in at least 2 annual surveillances. An increase in non-PCV7 serotypes with antibiotic resistance, beyond 6A and 19A, occurred. Intermediate resistance to penicillin was observed in serotype 23B, 15B, 15C, 15A, 35F, 6C, and 24F pneumococci. Their exact role in invasive and non-invasive disease remains to be seen in the years ahead.

Rotavirus is the leading cause of acute gastroenteritis among young children worldwide. A prospective multi-center study was conducted (2007-2008) in five Pediatric Hospitals to determine the prevalence, the clinical characteristics, and genotype distribution of rotavirus infection in Greece. Faecal samples were examined for the presence of group A rotavirus antigen by immunochromatography. Rotavirus strains were subjected to G and P genotyping by reverse-transcriptase polymerase chain reaction (PCR) and sequencing. A total of 393 children (216 boys) of median age 23 months, participated in the study. Rotavirus was the cause of acute gastroenteritis in 166 children, 42.3% (CI 95%, 37.4-47.1%) of non-hospitalized and 47.8% (CI 95%, 41.7-53.9%) of hospitalized patients. Rotavirus gastroenteritis occurred between December and April in 78.6% of the cases. Most children with RVG (77.8%) were between 3 months and 3 years old. The mean value of Clark severity score was 12.9 +/- 5.1 for RVG and 10.5 +/- 4.9 for non-RVG (P < 0.01). Genotypes were determined in 117 strains and their distribution was as following: G1P[8], 49%; G2P[4], 31%; G4P[8], 10%; G9P[8], 9%; and G8P[14], 1%. In conclusion, rotavirus is a frequent cause of acute gastroenteritis in Greece. The genotypes circulating are similar with those of other European countries. J Med Virol

The purpose of this study was to assess the complex interaction of the heptavalent pneumococcal conjugate vaccine (PCV7) and individual classes of antimicrobial agents on the nasopharyngeal carriage of erythromycin-resistant pneumococci within the day-care center population. Between February 2005 and May 2007, nasopharyngeal cultures for Streptococcus pneumoniae were obtained from 1,829 day-care center attendees in Central Greece. Thirty-one percent of the pneumococci were erythromycin-resistant; 85.2% of these isolates were also penicillin-nonsusceptible. PCV7 immunization was associated with decreased carriage of erythromycin-resistant PCV7 serotypes but not with an overall decrease in erythromycin-resistant S. pneumoniae colonization. The largest decline in the carriage of erythromycin-resistant pneumococci, particularly of PCV7 serotypes, was observed among vaccinated attendees who had not been exposed to antimicrobials within the preceding 3 months. Exposure to macrolides, 90% clarithromycin, significantly correlated with erythromycin resistance (adjusted odds ratio [AOR] = 2.08, 95% confidence interval [CI] = 1.38-3.12). There was a trend toward an association between the use of oral cephalosporins, other than cefprozil and cefaclor, and colonization with erythromycin-resistant pneumococci (AOR = 1.91, 95% CI = 0.92-3.96). Penicillins had a nonsignificant correlation with the carriage of erythromycin-resistant pneumococci (AOR = 1.17, 95% CI = 0.80-1.71). Despite the widespread PCV7 immunization, the antibiotic pressure, particularly of macrolides, continues to cause dissemination of erythromycin-resistant, commonly multidrug-resistant, pneumococci within the day-care center population.

Short course antimicrobial therapy is suggested for group A streptococcal tonsillopharyngitis.The bacteriologic and clinical efficacies of clarithromycin [30 or 15 mg/kg/day twice daily (b.i.d.)] or amoxicillin/clavulanate (43.8/6.2 mg/kg/day b.i.d.) for 5 days or penicillin V (30 mg/kg/day 3 times a day) for 10 days were compared. In a randomized, open label, parallel group, multicenter study, 626 children (2-16 years old) with tonsillopharyngitis were enrolled; 537 were evaluable for efficacy. Follow-up evaluations were performed at 4-8 and 21-28 days after therapy.At enrollment, 26% of the Streptococcus pyogenes isolates were clarithromycin-nonsusceptible. All regimens had an apparently similar clinical efficacy. The long term S. pyogenes eradication rates were 102 of 123 (83%) with amoxicillin/clavulanate and 88 of 114 (77%) with penicillin V. In the 30- and 15-mg/kg/day clarithromycin groups, eradication occurred in 71 of 86 (83%) and 59 of 80 (74%) of the clarithromycin-susceptible isolates (P = 0.33), and in 4 of 28 (14%) and 5 of 26 (19%) of the clarithromycin-resistant isolates, respectively (clarithromycin-susceptible versus -resistant, P0.0001). Both clarithromycin dosages were well-tolerated.In group A streptococcal tonsillopharyngitis, 5 days of clarithromycin or amoxicillin/clavulanate treatment had clinical efficacy comparable with that of 10 days of penicillin V treatment; however, amoxicillin/clavulanate and penicillin V were bacteriologically more effective than clarithromycin because of its failure to eradicate the clarithromycin-resistant S. pyogenes isolates. The 5-day clarithromycin regimens are not recommended for treatment of streptococcal tonsillopharyngitis in areas where in vitro resistance of group A streptococci to clarithromycin is common.

Streptococcus pyogenes isolates with the same pulsed-field patterns were recovered from the throat cultures of a child with tonsillopharyngitis before and after treatment with miocamycin, a 16-membered macrolide. The initial isolate was macrolide-susceptible, but the isolates after the treatment were resistant to 14 and 15-membered macrolides and had two amino acid (65WR66) deletions in ribosomal protein L4.

A total of 128 Streptococcus pneumoniae isolates that were susceptible to penicillin but resistant to non-β-lactam agents were isolated from young carriers in Greece and analyzed by antibiotic susceptibility testing, serotyping, restriction fragment end labeling (RFEL), and antibiotic resistance genotyping. The serotypes 6A/B (49%), 14 (14%), 19A/F (11%), 11A (9%), 23A/F (4%), 15B/C (2%), and 21 (2%) were most prevalent in this collection. Of the isolates, 65% were erythromycin resistant, while the remaining isolates were tetracycline and/or trimethoprim-sulfamethoxazole resistant. Fifty-nine distinct RFEL types were identified. Twenty different RFEL clusters, harboring 2 to 19 strains each, accounted for 76% of all strains. Confirmatory multilocus sequence typing analysis of the genetic clusters showed the presence of three international clones (Tennessee 23F -4, England 14 -9, and Greece 6B -22) representing 30% of the isolates. The erm (B) gene was present in 70% of the erythromycin-resistant isolates, whereas 18 and 8% contained the mef (A) and mef (E) genes, respectively. The pneumococci representing erm (B), erm (A), and mef genes belonged to distinct genetic clusters. In total, 45% of all isolates were tetracycline resistant. Ninety-six percent of these isolates contained the tet (M) gene. In conclusion, penicillin-susceptible pneumococci resistant to non-β-lactams are a genetically heterogeneous group displaying a variety of genotypes, resistance markers, and serotypes. This suggests that multiple genetic events lead to non-β-lactam-resistant pneumococci in Greece. Importantly, most of these genotypes are capable of disseminating within the community.

We describe the first case of Kingella kingae arthritis in a 16-month-old girl in Greece, which has been diagnosed by novel molecular techniques. A joint aspiration of her knee was performed before the initiation of antibiotics, as well as on the 5th and 14th day of empiric antimicrobial therapy. The synovial fluid white blood cell count decreased from 65,000 to 1500 cells/mm, but the percentage of neutrophils remained 90% in all 3 specimens. Molecular analysis of the synovial fluid specimens by real-time polymerase chain reaction and multilocus sequence typing enabled us to reveal the presence of K. kingae belonging to the international sequence type-complex 14, which persisted up to the fifth day of antibiotic therapy.

Background In Greece recently, higher-valent pneumococcal conjugate vaccines (PCVs) replaced the 7-valent (PCV7); the 10-valent (PCV10) became available in May 2009 and the 13-valent (PCV13) in June 2010. Methods We investigated the nasopharyngeal colonization with Streptococcus pneumoniae in day-care center attendees in Athens and the prefecture of Viotia. Between December 2010 and June 2011, nasopharyngeal cultures were obtained 4 times, at enrollment and then every 6 to 8 weeks. Results Among the 233 children, 225 (96.6%) had been vaccinated with ≥1 dose of PCV7. One tenth of the PCV7 vaccinated attendees had also received ≥1 dose of PCV13 or PCV10. During the 4 samplings, 358 isolates were recovered from a total of 874 samples. Of the 233 children, 183 (78.5%) were found to carry S. pneumoniae at least once. The overall serotype distribution among carriers was similar regardless of the time lapsed since the last PCV7 dose. A high frequency of 19A (17.1%) coincided with a low frequency of 19F (1.4%). Non-PCV13 serotypes accounted for 73.1% of the isolates; 23B, 15B/C, 16F, 21, 11A, 15A, 6C, 10A, 22F and 23A were the most common. Among attendees aged 24–59 months (median age 42 months), prolonged carriage of a non-PCV13 serotype was relatively common, mainly for 21 and 16F. One out of 4 cases of colonization with the prevalent non-PCV13 serotypes was followed by persistent carriage for 5 to 14 weeks. Conclusions During this period of transition to the higher-valent PCVs in the day-care center setting, non-PCV13 serotypes dominated and exhibited prolonged colonization. The frequency and the duration of prolonged carriage tends to be increased, if sampling frequency increases and the carriage time before and after positive cultures is taken into consideration. Further studies regarding the fitness of the colonizing non-PCV13 serotypes will likely to be seen in the future.

BACKGROUND Macrolide resistance among Streptococcus pyogenes strains is increasing in many European countries. Greece was not considered a country with high prevalence of macrolide-resistant S. pyogenes strains, and until now the genetic mechanism of resistance was unknown. METHODS During the 25-month period from December, 1998, to December, 2000, pharyngeal cultures for S. pyogenes were performed on 743 Greek children with the clinical diagnosis of pharyngitis. The children were 1 to 16 years old (median age, 7 years) and were living in Central and Southern Greece. S. pyogenes isolates were tested for their susceptibility to erythromycin, clarithromycin, azithromycin, clindamycin, penicillin G, amoxicillin/clavulanate and cefprozil. The erythromycin-resistant isolates were further studied for their genetic mechanism of resistance by means of PCR. RESULTS Of a total of 275 S. pyogenes isolates recovered, 105 (38%) were erythromycin-resistant (MIC > or = 1 microgram/ml) [corrected], with 54, 45 and 1% of them carrying mef(A), erm(A) [subclass erm(TR)] and erm(B) gene, respectively. The prevalence of erythromycin-resistant strains was 29 and 42% during the time periods December, 1998, to December, 1999, and January, 2000, to December, 2000, respectively. All erythromycin-resistant isolates were also resistant to clarithromycin and azithromycin. The isolates carrying the erm(A) gene were inducibly resistant to clindamycin. The 275 S. pyogenes isolates had ceprozil MICs < or = 0.032 microgram/ml. CONCLUSIONS The current high (38%) prevalence of erythromycin-resistant S. pyogenes in Central and Southern Greece requires continuous surveillance and careful antibiotic policy.

In January 1996 the emergence of penicillin-susceptible, multiresistant serotype 6B Streptococcus pneumoniae isolates resistant to chloramphenicol, tetracycline, erythromycin, clindamycin and trimethoprim-sulfamethoxazole was observed in young carriers in the city of Patras, located in the southwestern region of Greece. Later, a significant spread of pneumococci with this unusual phenotype was noted in carriers living in various other areas of the country. Using restriction fragment length polymorphism of the ribosomal RNA genes, clonal relationships were found between these Greek strains and serotype 6B penicillin-susceptible, multiresistant pneumococci isolated in France between January 1992 and September 1996. The French and Greek isolates appear to have a common ancestry.

A total of 145 penicillin-nonsusceptible Streptococcus pneumoniae strains were isolated from young carriers in Greece and analyzed by antibiotic susceptibility testing, serotyping, restriction fragment end labeling (RFEL), and penicillin-binding protein (PBP) genotyping. The serotypes 23A and 23F (54%), 19A and 19F (25%), 9V (5%), 15A, 15B, and 15C (4%), 6A and 6B (4%), and 21 (4%) were most prevalent in this collection. Fifty-three distinct RFEL types were identified. Sixteen different RFEL clusters, harboring 2 to 32 strains each, accounted for 82% of all strains. Eight of these genetic clusters representing 60% of the strains were previously identified in other countries. A predominant lineage of 66 strains (46%) harboring five RFEL types and the serotypes 19F and 23F was closely related to the pandemic clone Spain 23F -1 (genetic relatedness of ≥85%). Another lineage, representing 11 strains, showed close genetic relatedness to the pandemic clone France 9V -3. Another lineage of 8 serotype 21 strains was Greece specific since the RFEL types were not observed in an international collection of 193 genotypes from 16 different countries. Characterization of the PBP genes pbp1a , pbp2b , and pbp2x revealed 20 distinct PBP genotypes of which PBP type 1-1-1, initially observed in the pandemic clones 23F and 9V, was predominantly present in 11 RFEL types in this Greek collection of penicillin-nonsusceptible strains (55%). Sixteen PBP types covering 52 strains (36%) were Greece specific. This study underlines the strong contribution of penicillin-resistant international clones to the prevalence and spread of penicillin-nonsusceptible pneumococci among young children in Greece.

In 1996, 19 isolates of serotype 6B Streptococcus pneumoniae with a unique resistance pattern were found in carriers attending daycare centres in Patras, Southwestern Greece. These isolates were penicillin susceptible but resistant to chloramphenicol, tetracycline, erythromycin, clindamycin and trimethoprim-sulphamethoxazole. Subsequently, isolates with the same characteristics were found in 23 additional carriers in central and southern Greece in 1997–98 as well as in 19 carriers in central Italy in 1997, and in seven carriers in southern Israel in 1998. Carriers were all children under 6 years of age, attending daycare centres or outpatient hospital visits. The relatedness of the isolates was determined on representative isolates from the three countries by pulsed-field gel electrophoresis of Sma I digests of chromosomal DNA. Most Greek isolates were identical to each other, while isolates from Italy and Israel showed one to three band differences, with all isolates being closely related to each other as well as to the isolates from Greece. We have therefore documented the presence of this unique clone of S. pneumoniae in these three countries and have named this the ‘Mediterranean' clone. While isolates appear to have a common origin, their source and direction of spread are unknown. However, isolates from Italy showed the most diversity, suggesting that this clone had been present in that country for a longer period than it had been in Greece.

We evaluated nasopharyngeal colonization with erythromycin-resistant Streptococcus pneumoniae during the first 2 years of life in central and southern Greece. Of 2448 children studied from February 1997 to February 1999, 766 (31%) carried 781 pneumococcal isolates. Ninety-five (3.9%) of the children attended day care centers. Eighteen percent of the pneumococci were resistant to erythromycin (minimal inhibitory concentration 1 to >128 microg/mL), with 67.9% of them carrying the erm(B) gene and 29.2% mef(A) gene products. Four strains possessed neither the erm(B) nor the mef(A) gene. Multidrug resistance occurred in 97% and 40% of isolates carrying the erm(B) and mef(A) gene, respectively. An association was found between the erm(B) gene and serotypes 6B and 23F and between the mef(A) gene and serotypes 14 and 19F. A significant relationship existed between carriage of erythromycin-resistant pneumococci and use of macrolides or beta-lactams in the previous 3 months; the association was strongest when macrolide therapy was administered during the last month (odds ratio, 5.92; P=.0001). The findings indicate the necessity of a judicious use of both macrolides and beta-lactams in young children to reduce the colonization with erythromycin-resistant pneumococci and the subsequent spread of such strains to the community.

The prevalence, resistance patterns and serotypes of antibiotic-resistant Streptococcus pneumoniae strains recovered from Greek carriers under 24 months of age were studied. From February 1997 to April 1998, nasopharyngeal cultures were performed in 1,269 children (ages 2-23 months, median 11 months) living in various areas of central and southern Greece. Resistance (including both intermediate and resistant isolates) to one or more antimicrobial agents was found in 132 of the 421 (31%) Streptococcus pneumoniae isolates, as follows: penicillin, 9% intermediate, 7.6% resistant; cefotaxime, 5.2% intermediate, 0.5% resistant; erythromycin, 0.7% intermediate, 18.1% resistant; clindamycin, 0.2% intermediate, 12.4% resistant; tetracycline, 0.7% intermediate, 16.4% resistant; chloramphenicol, 12.4% resistant; and trimethoprim-sulfamethoxazole, 3.8% intermediate, 14.3% resistant. The MICs of penicillin for 66% of the penicillin-nonsusceptible pneumococci were 1-4 microg/ml. Multidrug resistance was found in 64% of penicillin-nonsusceptible and 37% of penicillin-susceptible strains. Sixty-two percent of the penicillin-susceptible, multidrug-resistant strains belonged to serotype 6B and were resistant to all five non-beta-lactam agents tested. This notable serotype 6B resistance pattern was described for the first time in a previous study performed from December 1995 to February 1996 in the city of Patras, southwestern Greece. Seventy-two percent of antibiotic-resistant isolates belonged to serotypes 6B, 9V, 14, 18C, 19F and 23F. These results document the spread of resistant pneumococcal strains in central and southern Greece, many of which are multidrug resistant.

Black hairy tongue (BHT) is an unusual condition in adults, and is characterised by marked accumulation of keratin on the filiform papillae of the dorsum of the tongue resulting in a hairlike appearance. The colour of the papillae may vary from brown to black. We describe a case of BHT in a 2-month-old infant. An extended review of the literature suggests that our case is the youngest ever reported. In conclusion, although BHT is considered benign, clinical, haematological and histological, evaluation is recommended to exclude several entities which can present as pigmented lesions of the oral mucosa.

The resistance to beta-lactam and non-beta-lactam antibiotics of 133 nasopharyngeal isolates of Streptococcus pneumoniae recovered from December 1995 to February 1996 from children attending seven day-care centers in southwestern Greece was studied. Reduced susceptibility to one or more anti-microbial agents was found in 70 isolates (53%), as follows: penicillin, 17% intermediate, 12% resistant; cefotaxime, 10.5% intermediate, 1.5% resistant; trimethoprim-sulfamethoxazole, 8% intermediate, 35% resistant; chloramphenicol, 27% resistant; tetracycline, 29% resistant; and erythromycin/clindamycin, 19% resistant. Eighty-seven percent of penicillin-intermediate or -resistant strains belonged to serogroups/serotypes 19, 21, and 23. Fifty-six percent of the antibiotic-resistant pneumococci were multiply resistant, including serogroup 6 strains that were penicillin-susceptible but resistant to all non-beta-lactam drugs tested, as well as serogroup 23 strains resistant to penicillin, chloramphenicol, tetracycline, and trimethoprim-sulfamethoxazole. The high incidence of antibiotic-resistant pneumococci and the divergent and unique resistance patterns found in this study underline the need for global surveillance of S. pneumoniae to document the evolution and spread of resistant strains and to guide therapy.

We studied the attitudes with regard to occupational vaccines and vaccination coverage among healthcare workers in pediatric departments. Completed vaccination rates were 33%, 33%, 41.7%, 3%, 5.8%, 69.2% and 36.3% against measles, mumps, rubella, varicella, hepatitis A, hepatitis B and tetanus-diphtheria, respectively. Susceptibility rates were 14.2%, 15.7%, 14.6%, 7.6%, 87.4%, 22.6% and 61.8% for measles, mumps, rubella, varicella, hepatitis A, hepatitis B and tetanus-diphtheria, respectively. Mandatory vaccinations were supported by 70.6% of healthcare workers, with considerable differences by target disease.

Introduction In Greece, fusidic acid and clindamycin are commonly used for the empiric therapy of suspected staphylococcal infections. Methods The medical records of children examined at the outpatient clinics or admitted to the pediatric wards of the University General Hospital of Larissa, Central Greece, with community-associated staphylococcal infections from January 2003 to December 2009 were reviewed. Results Of 309 children (0-14 years old), 21 (6.8%) had invasive infections and 288 (93.2%) skin and soft tissue infections (SSTIs). Thirty-five patients were ≤30 days of age. The proportion of staphylococcal infections caused by a community-associated methicillin-resistant Staphylococcus aureus (CA-MRSA) isolate increased from 51.5% (69 of 134) in 2003-2006 to 63.4% (111 of 175) in 2007-2009 (P = 0.037). Among the CA-MRSA isolates, 88.9% were resistant to fusidic acid, 77.6% to tetracycline, and 21.1% to clindamycin. Clindamycin resistance increased from 0% (2003) to 31.2% (2009) among the CA-MRSA isolates (P = 0.011). Over the 7-year period, an increase in multidrug-resistant CA-MRSA isolates was observed (P = 0.004). One hundred and thirty-one (93.6%) of the 140 tested MRSA isolates were Panton-Valentine leukocidin-positive. Multilocus sequence typing of 72 CA-MRSA isolates revealed that they belonged to ST80 (n = 61), ST30 (n = 6), ST377 (n = 3), ST22 (n = 1), and ST152 (n = 1). Resistance to fusidic acid was observed in ST80 (58/61), ST30 (1/6), and ST22 (1/1) isolates. Conclusion In areas with high rate of infections caused by multidrug-resistant CA-MRSA isolates, predominantly belonging to the European ST80 clone, fusidic acid and clindamycin should be used cautiously as empiric therapy in patients with suspected severe staphylococcal infections.

Objectives: Immature gut motility in very low birth weight infants causes feeding intolerance. We evaluated the effect of domperidone, a prokinetic agent, on gastric emptying in very low birth weight infants. Methods: The study was conducted in a crossover design. Gastric emptying was assessed ultrasonographically by measuring the time it took the antral cross sectional area of the stomach to reach half of the value of the 1st measurement on 2 occasions: (1) upon administration of domperidone (0.3 mg/kg/8 h p.o.) for at least 2 days and (2) while patients received an equal quantity of sterile water. 11 infants were randomly assigned to receive domperidone before the 1st measurement and the remaining afterwards. There was a 3–5 day interval between the 2 measurements. At the time when both measurements were performed, every neonate had been receiving the same quantity and quality of milk. 12 infants were formula-fed while 10 were fed their own mother’s supplemented milk. Results: 22 infants with a mean (SD) birth weight of 1,377 g (319) and a mean (SD) gestational age of 30.2 weeks (2.1) were studied. The mean (SD) antral cross sectional area half-value time (in minutes) was 47.6 (23.9) in the domperidone group and 68.2 (25.5) in the control group (p = 0.008). There was no significant difference between formula-fed neonates and neonates fed their own mother’s milk in either group. Conclusions: Domperidone significantly reduces gastric emptying in preterm neonates, and this may account for its effect in cases of disturbances related to gut motility.

In Greece, the heptavalent pneumococcal conjugate vaccine (PCV7) became available in October 2004 and it was incorporated into the national immunization schedule in January 2006.In February 2005, a yearly surveillance of the nasopharyngeal colonization with Streptococcus pneumoniae in children attending day-care centers in Central Greece began.Between February 2005 and May 2007, nasopharyngeal cultures were obtained from 1829 children aged 13-76 months (median age, 47 months). The proportion of attendees vaccinated withor =1 doses of PCV7 increased from 13% (2005) to 33% (2006) and to 70% (2007); 98% had been immunized on toddler catch-up schedules. Among vaccinated carriers, the proportion of PCV7 serotypes decreased from 33% (2005) to 29% (2006) and to 8.6% (2007) (chi for trend, P0.001), the proportion of PCV7-related serotypes increased from 13% (2005) to 26% (2006) and to 28% (2007) (P = 0.16), whereas the proportion of non-PCV7 serotypes was 48% in 2005, 31% in 2006, and 55% in 2007 (P = 0.17). The proportion of PCV7 serotypes declined also among unvaccinated carriers. The carriage of serotype 19A did not increase. Among vaccinated carriers, the rate of highly penicillin-resistant isolates decreased from year 1 to year 3, respectively, 11%, 7.7%, and 0.6% (P = 0.001), whereas the proportion of penicillin-intermediate pneumococci was 13% in 2005, 23% in 2006, and 26% in 2007 (P = 0.22).In Central Greece, widespread PCV vaccination was followed by a significant reduction of carriage of highly penicillin-resistant pneumococci. The frequency of penicillin-intermediate isolates did not change significantly among vaccinated carriers.

Since the late 1990s, the prevalence of erythromycin-resistant Streptococcus pyogenes has significantly increased in several European countries. Between January 1999 and December 2002, 1,577 isolates of S. pyogenes were recovered from children with tonsillopharyngitis living in various areas of Western Greece. Erythromycin resistance was observed in 379 (24%) of the 1,577 isolates. All erythromycin-resistant strains along with 153 randomly selected erythromycin-susceptible S. pyogenes isolates were tested for their antimicrobial susceptibility, resistance phenotypes, and genotypes. Representative isolates underwent emm gene sequence typing. Isolates with reduced susceptibility to telithromycin (MIC, ≥2 μg/ml) were studied for multilocus sequence type, L22, L4, and 23S rRNA mutations. Of the total 379 erythromycin-resistant isolates, 193 (50.9%) harbored the mef (A) gene, 163 (43%) erm (A), 1 (0.3%) mef (A) plus erm (A), and 22 (5.8%) the erm (B) gene. Among the erythromycin-susceptible isolates, emm 1 (25%), emm 2 (12.5%), and emm 77 (12.5%) predominated. Furthermore, among the erythromycin-resistant isolates, emm 4 (30.6%), emm 28 (22.2%), and emm 77 (12.5%) prevailed. Resistance to telithromycin was observed in 22 (5.8%) of the erythromycin-resistant isolates. Sixteen (72.7%) of the 22 isolates appeared to be clonally related, since all of them belonged to emm type 28 and multilocus sequence type 52. One of the well-known mutations (T2166C) in 23S rRNA, as well as a new one (T2136C), was detected in erythromycin- and telithromycin-resistant isolates. High incidence of macrolide resistance and clonal spread of telithromycin resistance were the characteristics of the Greek S. pyogenes isolates obtained from 1999 to 2002.

The outer membrane proteins UspA1 and UspA2 are candidate antigens for a Moraxella catarrhalis vaccine. We previously reported that 103 of 108 isolates (95%) from young children expressed UspA1 detected by reactivity with the monoclonal antibody mAb24B5. The aim of the present study was to investigate mechanisms controlling UspA1 expression by analysis of five mAb24B5 non-reactive isolates. Four of these strains were characterized by (i) decreased or absent transcription of uspA1 and uspA2 and (ii) clustered mutations and deletions in the promoter region of both uspA1 and uspA2. Antigenic or phase variation were not responsible for reduced levels of UspA1 expression. While mAb24B5-positive isolates expressing normal levels of uspA1 and uspA2 mRNA belonged to the previously described 16S rRNA type 1 phylogenetic group, these four mAb24B5-negative isolates were found to belong to the 16S rRNA gene types 2 or 3. The remaining mAb24B5-negative isolate (#610) belonged to 16S rRNA type 1 and exhibited a posttranscriptional defect of UspA1 expression defined by normal levels of uspA1 mRNA and both recombinant and in vitro expression of mAb24B5-reactive UspA1. In conclusion, M. catarrhalis clinical isolates exhibiting reduced expression of UspA1 and UspA2 belonged to a distinct phylogenetic subpopulation. A UspA-based vaccine is unlikely to be effective against such isolates.

Erythromycin-resistant Streptococcus pneumoniae isolates from young carriers were tested for their antimicrobial susceptibility; additionally, inducibility of macrolide and clindamycin resistance was investigated in pneumococci carrying erm (A), erm (B), or mef (A). Of 125 strains tested, 101 (81%) were multidrug resistant. Different levels of induction were observed with erythromycin, miocamycin, and clindamycin in erm (B) strains; however, in erm (A) strains only erythromycin was an inducer. Induction did not affect macrolide MICs in mef (A) strains.

UspA1 and UspA2 of Moraxella catarrhalis are vaccine candidates. The aims of this study were to determine: (1) the frequencies of occurrence and (2) the degrees of conservation of two surface-exposed epitopes of the uspA1 and uspA2 genes and their respective gene products in 108 nasopharyngeal isolates from young children. The uspA1 and uspA2 genes were detected in 107 (99%) and 108 (100%) isolates, respectively. Twenty-three of 108 uspA2 genes (21%) were identified as the variant gene uspA2H. One-hundred and five isolates (97%) expressed the mAb17C7-reactive epitope shared by UspA1 and UspA2, and 103 isolates (95%) reacted with the UspA1-specific mAb24B5. The only isolate which lacked a uspA1 gene demonstrated reduced adherence to HEp-2 cells and complement sensitivity. The data indicate that both uspA genes and the expression of at least two surface-exposed epitopes are virtually ubiquitous in isolates from a population at risk for otitis media. A vaccine capable of inducing a bactericidal immune response against the mAb17C7- and/or mAb24B5-reactive epitopes appears promising.

The serotype distribution of 781 nasopharyngeal pneumococcal isolates recovered from 2448 unselected children aged 2-23 months was studied. Only 3.9% of the children for whom cultures were performed attended day care centers. The proportions of pneumococcal isolates that belonged to serotypes related to the 7-, 9- and 11-valent conjugate pneumococcal vaccine were 65%, 66%, and 70%, respectively. The pneumococcal carriage rate among untreated children was 34%; the rates among children treated with antibiotics during the periods 1-30 or 31-60 days before the time of nasopharyngeal sampling were 25% and 36%, respectively. There was a significant positive association between antimicrobial use and carriage of antibiotic-resistant pneumococci, which belonged mainly to vaccine-related serotypes. The proportion of isolates that belonged to vaccine-related serotypes in untreated carriers was 72%; however, the proportions in carriers treated 1-30 days or 31-60 days before sampling were 66% and 56%, respectively. In the nasopharynx, antimicrobial use selects for antibiotic-resistant pneumococci, mainly of vaccine-related serotypes, whereas it may promote an increase in the frequency of colonization with nonvaccine serotypes.