Propranolol in the therapeutic strategy of infantile laryngotracheal hemangioma: A preliminary retrospective study of French experience

Objective

Preliminary assessment of the efficacy of propranolol on subglottic hemangioma in children on a nation-wide scale.

Methods

Multicentric, retrospective study of clinical files of 14 children; pre- and post-treatment endoscopies.

Results

Mean age at diagnosis was 2.3 (0.7-4) months. Mean percentage of airway obstruction was 68% (15-90) before propranolol introduction. Propranolol was started at 5.2 (0.7-16) months of age. This treatment was effective in all cases with a mean regression of the stenosis to 22% after 2 weeks and 12% after 4 weeks. Other medical treatments (steroids) could be stopped. In one patient, a side effect of propranolol motivated the switch to another β-blocker. In four patients, treatment was stopped after 5.2 (1-10) months with a relapse in 2 (50%) cases. One of these two patients developed a resistance to propranolol and required a surgical procedure by external approach.

Conclusion

This preliminary nation-wide survey confirms propranolol high effectiveness against airways’ localization of infantile hemangiomas. Propranolol also allows alleviation or cut-off of previous medical treatments. However, recurrences are possible after early treatment interruption.     

Mouse model of Aspergillus and Alternaria induced rhinosinusitis

Objective

Fungi are known to induce the production of chemical mediators from respiratory epithelial cells and have been increasingly recognized as important pathogens in sinusitis. However, the exact role of fungi in the pathogenesis of rhinosinusitis has not been clearly established. This study was performed to improve our understanding of the role of fungi in the pathogenesis of rhinosinusitis by developing an animal model of fungus induced rhinosinusitis.

Methods

Fifty mice (C57BL6/J) were divided into five groups. Sham-operated group was the first group. In the second group, Aspergillus versicolor (Group IIa) and Alternaria alternata (Group IIb) (10⁶ spores/ml) were inoculated into the nasal cavity. In the third group, fungi were inoculated into the nasal cavity in the presence of mucosal scratch (Group IIIa,b) and the fourth group was a nasal mucosal scratch only (Group IV). The fifth was a negative control (Group V). The fungi were inoculated once a week on six occasions and then the animals were sacrificed at 7 weeks. The histological sections were examined in a blind manner for the appearance of neutrophil clusters and epithelial thickness with hematoxylin–eosin stain, and mucus secreting glands using the Alcian blue/periodic acid Schiff stain.

Results

Non-invasive fungal sinusitis had been induced with increased numbers of neutrophil clusters after Aspergillus and Alternaria exposure. The mice with the mucosal scratch wounds had significantly more inflammatory cell infiltration and epithelial thickening; but eosinophils were not commonly found. The mice with fungal sinusitis had goblet cell hyperplasia and increased mucus secretion in the sinonasal cavity.

Conclusions

Inoculation of fungi in the nasal cavity induced rhinosinusitis in C57BL6/J mice. This mouse model may be used for better understanding of the role of fungi in the pathogenesis of rhinosinusitis.     

Chlamydia pneumoniae and typical bacteria occurrence in adenoid in children qualified for adenoidectomy

Objective

The ability to occur in a persistent form and to inhibit apoptosis in host cells are the features of Chlamydia pneumoniae which may be associated with adenoid hypertrophy.This study aimed to (1) demonstrate the presence of C. pneumoniae DNA in the adenoid in children qualified for adenoidectomy, (2) evaluate the correlation between the presence of C. pneumoniae in the adenoid and the child's age, the size of adenoid, the incidence of recurrent respiratory tract infections, (3) demonstrate the co-occurrence of C. pneumoniae and typical bacteria in the adenoid tissue.

Methods

A group of 200 children aged from 2 to 16 years (mean age 6.4) undergoing adenoidectomy from February 2010 to May 2011 were enrolled to the study. Adenoid tissue removed during planned adenoidectomy was analyzed for the presence of C. pneumoniae by real-time PCR and for the presence of typical bacteria by bacteriological culture. Statistical analysis was conducted using SPSS PASW Statistics 18 software.

Results

C. pneumoniae was detected in the adenoid in 5.5% of the 200 children. Positive results were obtained most frequently (24.1%, 7/29) in the eldest group (10–16 years). In PCR positive children the degree of choanal narrowing by an adenoid tended to be larger than in PCR negative children. The occurrence of C. pneumoniae in the adenoid was not associated with a presence of a specific typical bacterium.

Conclusions

The presence of C. pneumoniae in an adenoid was most frequently detected in the children of 10–16 years qualified for adenoidectomy.     

Haemophilus influenzae and Haemophilus haemolyticus in tonsillar cultures of adults with acute pharyngotonsillitis

Objective

The aim of this study was to evaluate the clinical implication of Haemophilus haemolyticus, one of the closest relative of Haemophilus influenzae, on acute pharyngotonsillitis.

Methods

We applied polymerase chain reaction (PCR) for 16S ribosomal DNA (rDNA) and IgA protease gene (iga) to distinguish H. haemolyticus and H. influenzae.

Results

Among the 199 Haemophilus spp. isolated from 214 patients with acute pharyngotonsillitis, 52 (24.3%) H. influenzae strains and 23 (10.7%) H. haemolyticus strains were identified by polymerase chain reaction (PCR) for 16S rDNA and IgA protease gene (iga). All H. haemolyticus strains showed hemolysis on horse blood agar and there were no other Haemophilus spp., nonhemolytic H. haemolyticus and H. influenzae variant strains that had absent iga gene. H. hemolyticus showed close genetic relationship with H. influenzae evaluated by pulsed field gel electrophoresis (PFGE). The cases of acute pharyngotonsillitis showing WBC = 7000/mm³ or CRP = 8 mg/dl were frequently found among cases with H. influenzae rather than cases with H. haemolyticus.

Conclusion

H. haemolyticus is a pharyngeal commensal that is isolated frequently from adults with acute pharyngotonsillitis.     

Postcricoid infantile hemangioma: Management of a case in the propranolol era

We describe a rare case of a postcricoid infantile hemangioma (IH) and treatment with propranolol. A newborn female was found to have a large, postcricoid submucosal mass. The patient was brought to the operating theater for endoscopy and biopsy. The histopathology and GLUT-1 immunoreactivity confirmed the diagnosis of IH. The lesion was treated with direct injection of triamcinolone (40 mg/ml) and systemic propranolol therapy. The patient responded well, and her symptoms resolved quickly. Propanolol provides a promising therapy as a first line treatment to avoid surgery, tracheostomy, and prolonged use of steroids.     

De novo dominant mutation of SOX10 gene in a Chinese family with Waardenburg syndrome type II

Objective

Waardenburg syndrome is a rare genetic disorder, inherited as an autosomal dominant trait. The condition is characterized by sensorineural hearing loss and pigment disturbances of the hair, skin, and iris. The de novo mutation in the SOX10 gene, responsible for Waardenburg syndrome type II, is rarely seen. The present study aimed to identify the genetic causes of Waardenburg syndrome type II in a Chinese family.

Methods

Clinical and molecular evaluations were conducted in a Chinese family with Waardenburg syndrome type II.

Results

A novel SOX10 heterozygous c.259-260delCT mutation was identified. Heterozygosity was not observed in the parents and sister of the proband, indicating that the mutation has arisen de novo. The novel frameshift mutation, located in exon 3 of the SOX10 gene, disrupted normal amino acid coding from Leu87, leading to premature termination at nucleotide 396 (TGA). The high mobility group domain of SOX10 was inferred to be partially impaired.

Conclusion

The novel heterozygous c.259-260delCT mutation in the SOX10 gene was considered to be the cause of Waardenburg syndrome in the proband. The clinical and genetic characterization of this family would help elucidate the genetic heterogeneity of SOX10 in Waardenburg syndrome type II. Moreover, the de novo pattern expanded the mutation data of SOX10.     

Screening of SLC26A4, FOXI1 and KCNJ10 genes in unilateral hearing impairment with ipsilateral enlarged vestibular aqueduct

Objective

To investigate the implication of SLC26A4, FOXI and KCNJ10 genes in unilateral hearing impairment associated with ipsilateral inner ear malformation (Enlargement of the vestibular aqueduct and/or Mondini dysplasia).

Methods

We have gathered 25 patients presenting unilateral hearing impairment and ipsilateral enlarged vestibular aqueduct. For each of the patients, we have analyzed SLC26A4, FOXI1 and KCNJ10 genes sequences.

Results

The analysis of SLC26A4 revealed only eight heterozygous SLC26A4 sequence variants, three of them being novel (p.Met147Ile, p.Asn538Asn and p.Leu627Arg). None of the patients carried a second mutation on the other allele. Moreover, the SLC26A4 locus was excluded by segregation analysis in two families. No mutations were present in FOXI1 and KCNJ10 genes.

Conclusions

Together, these data suggest that SLC26A4, FOXI1 and KCNJ10 are not major determinants in unilateral deafness and enlarged vestibular aqueduct compared with their implication in Pendred syndrome and non-syndromic bilateral enlarged vestibular aqueduct.     

婴幼儿先天性声门下血管瘤的诊断和治疗

目的：探讨婴幼儿先天性声门下血管瘤的诊断和治疗。方法：9例患儿,反复喉喘鸣、喂哺困难及喉梗阻发作,电子支气管镜检查确诊为先天性声门下血管瘤。7例为声门下单侧病变,2例为双侧病变。其中8例采用气管切开后血管瘤内平阳霉素注射法进行治疗,术后患儿带金属气管套管出院,2周后门诊随访电子支气管镜复查,如果肿瘤未消失,再次行平阳霉素注射,直至肿瘤完全消失后进行堵管及拔管;1例单侧声门下血管瘤行瘤体吸割术,未行气管切开。结果：平阳霉素治疗的6例声门下单侧病变患者经过1次治疗后血管瘤完全消失,2例双侧病变患者经过2次治疗后血管瘤完全消失,8例均顺利堵管及拔管。1例行声门下血管瘤吸割术患者,术后2周复查黏膜光滑,完全修复。结论：对于体积较大,阻塞声门下腔范围较广的血管瘤,采用气管切开后血管瘤内平阳霉素注射法,能迅速解除气道梗阻,缩短自然病程,且戴管时间短,无术后声门下狭窄,是一种安全有效的治疗措施。对于体积较小的声门下血管瘤,在麻醉配合良好的情况下,行吸割术并给予良好止血,可避免气管切开,起到微创效果。     

Lack of Chlamydophila pneumoniae and predominance of Alloiococcus otitidis in middle ear fluids of children with otitis media with effusion

Objective

To investigate the presence of Chlamydophila pneumoniae and other bacterial pathogens in middle ear effusion samples obtained from children with otitis media with effusion (OME).

Materials and methods

Twenty-eight children (mean age 7.03; standard deviation 2.18) with OME unresponsive to medical therapy were included in the study. All of the children underwent ventilation tube insertion under general anesthesia. Eighteen patients were bilaterally affected whereas 10 children had unilateral disease. The middle ear fluids (46 samples in total) were collected during ventilation tube insertion, and were evaluated subsequently for the presence of C. pneumoniae and other bacterial pathogens using polymerase chain reaction (PCR).

Results

Although all samples were negative for C. pneumoniae, bacterial DNA was detected in 21 of 46 samples. Overall 40% of the patients (4/10) with unilateral involvement, and 61% of the patients (11/18) with bilateral involvement were positive for bacterial DNA. In 6 patients with bilateral OME bilateral samples were positive, whereas 5 patients with bilateral OME showed only unilateral positivity. According to the results of DNA sequencing analysis, all of the positive samples harbored only one bacterial species. In 12 of 46 samples Alloiococcus otitidis DNA (26%), in 7 Haemophilus influenzae DNA (15%), in one Streptoccoccus pneumoniae DNA (2%) and in one Moraxella catarrhalis DNA (2%) were present.

Conclusions

Our findings support that C. pneumoniae does not seem to have a role in OME in children whereas A. otitidis was found to be more frequent than the other common pathogens. Further studies are required to elucidate the exact pathogenetic role of these microorganisms in OME.     

Identification of both MT-RNR1 m.1555A>G and bi-allelic GJB2 mutations in probands with non-syndromic hearing loss

Objectives

Mutations in GJB2 and MT-RNR1 are common causes for non-syndromic sensorineural hearing loss (NSHL). In this study, we investigated the co-existence of both MT-RNR1 and bi-allelic GJB2 mutations in a large number of simplex and multiplex probands with NSHL.

Methods

485 simplex and 134 multiplex probands with NSHL were recruited for mutation screening of GJB2 and MT-RNR1 by bidirectional sequencing. Clinical features of probands with both MT-RNR1 and bi-allelic GJB2 mutations were further analyzed in comparison with extended family members.

Results

MT-RNR1m.1555A>G and bi-allelic GJB2 mutations were both detected in one (0.21%) simplex probands and two (1.49%) multiplex probands. Variable hearing phenotypes were found in a pair of siblings with both MT-RNR1 and bi-allelic GJB2 mutations.

Conclusion

MT-RNR1 and bi-allelic GJB2 mutations may co-exist not only in multiplex probands but also in simplex probands. The variable hearing phenotypes in closely-related family members may reflect the co-existence of different molecular causes and prompt extended genetic tests.     

Preliminary study of proinflammatory cytokines and chemokines in the middle ear of acute otitis media due to Alloiococcus otitidis

Alloiococcus otitidis is a newly discovered organism frequently detected in otitis media. However, the association of the organism with the development of otitis media has not been disclosed in detail yet. In the middle ear, proinflammatory cytokines and chemokines are released in association with infection by pathogens, and these cytokines contribute to the induction of an inflammatory reaction. To investigate the profile of inflammation-related cytokines in the acute phase of A. otitidis infection, we analyzed the release of proinflammatory cytokines and chemokines in middle ear effusions of acute otitis media due to A. otitidis, in comparison with acute otitis media due to the well-known Gram-positive middle ear pathogen Streptococcus pneumoniae. The amounts of proinflammatory cytokines (IL-8, IL-1beta, IL-6, TNF-alpha) and CXC chemokines (IP-10, I-TAC) were significantly increased in the A. otitidis group as well as in the S. pneumoniae group. Various inflammation-related cytokines/chemokines were induced in the A. otitidis-infected middle ear, and the profile of cytokines was very similar to that in S. pneumoniae infection. This preliminary study suggests that A. otitidis has the potential to induce these cytokines, contributing to the development of an inflammatory reaction in the middle ear cavity in a similar manner to S. pneumoniae.     

Identification of novel variants in the TMIE gene of patients with nonsyndromic hearing loss

Objective

To determine whether variants of the TMIE gene are causes of nonsyndromic deafness in Taiwan.

Methods

A genetic survey was made from 370 individuals, with 250 nonsyndromic hearing loss and 120 normal hearing individuals. Genomic DNA was extracted from peripheral blood leukocytes and then subjected to PCR to amplify selected exons and flanking introns of the TMIE gene; the amplified products were screened for base variants by autosequence. Data from the two groups were then compared using Fisher's two-tailed exact test and Armitage's trend test.

Results

The analysis revealed 7 novel variants in the TMIE gene. Of the 7 variants, 5 variants were found in both nonsyndromic hearing loss and normal hearing group. Both allelic and genotype frequencies of these sequence changes did not differ significantly between patients and controls (P > 0.05). However, a missense variant (c.257G > A) and one promoter variant (g.1-219A > T) were found in two patients with nonsyndromic hearing loss. Family study and microsatellite analysis found that c.257G > A variant is not inherited from his parents. The c.257G > A variant encodes a protein with glutamine at position 86 instead of arginine (p.R86Q), a residue that is conserved in mammals but different in fish, and predicted to be extracellular.

Conclusions

Despite the fact that the frequency of TMIE variants in our study subjects was low, we suggested that c.257G > A (p.R86Q) variant is a de novo and may be as a risk factor for the development of hearing loss in Taiwanese.     

MTHFR C677T and A1298C polymorphisms and risk of nonsyndromic orofacial clefts in a south Indian population

Objectives

Several lines of evidence suggest that the decrease in folate in periconceptional period or maternal use of folate antagonists has been associated with a higher risk of orofacial clefts (OFCs). MTHFR is a critical enzyme in folate metabolism that catalyzes the irreversible conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, thus playing a vital role in DNA synthesis and DNA methylation. The aim of our study was to determine whether there is any association between the susceptibility to Nonsyndromic cleft lip with or without cleft palate (NSCL/P) amongst the variations of MTHFR genotypes in South Indian population.

Methods

Our sample comprised 123 cases with NSCL/P and 141 controls without clefts or family history of clefting. The most common polymorphisms C677T (rs1801133) and A1298C (rs1801131) on the MTHFR gene were screened for the genotypes using PCR-RFLP.

Results

Both C677T and A1298C are polymorphic with minor allele frequencies of 0.131 and 0.429, respectively, for controls. Genotype data in control and cleft groups are following the Hardy Weinberg Equilibrium. There were no significant differences in genotypes of both polymorphisms between controls and NSCL/P. The pairwise LD values (D′ and r²) between C677T and A1298C are 1.0 and 0.096 respectively indicating no significant LD between these two SNPs. Haplotype phenotype analysis did not show the evidence for association. Gene–gene interaction showed the distribution of the observed combinations of the two MTHFR polymorphisms was not different between NSCL/P and controls (p = 0.887).

Conclusions

Our results do not support the hypothesis, that variants in the MTHFR gene confer a risk for NSCL/P in the South Indian population.     

Particular distribution of the GJB2/GJB6 gene mutations in Mexican population with hearing impairment

Background

Hereditary sensorineural hearing loss (SNHL) is a genetically heterogeneous disorder worldwide. Mutations in the GJB2 gene are a frequent cause of hereditary SNHL. There is a prevalence of certain mutations in various populations which suggests that specific mutations may be influenced by ethnic background.

Objective

To analyze the prevalence of GJB2, GJB6 mutations in several geographic areas of Mexico in patients with hereditary SNHL.

Materials and methods

One hundred and forty Mexican unrelated propositi with prelingual SNHL were included in the study. All patients had three previous generations born in Mexico and belonged to no specific ethnic group. Analyses of the GJB2 and GJB6 genes and mt.1555A < G were performed in all subjects.

Results

Twenty-three homozygous mutations, 57 heterozygous mutations, 1 double heterozygous (GJB2/GJB6) and 59 wild-type genotypes in the GJB2 gene were observed. Three patients had the homozygous c.del35 mutation whereas 26 patients were heterozygous for this gene defect. Only one patient with the GJB6 gene deletion was present (it includes the double heterozygous GJB2/GJB6). The mt.1555A > G mutation was not detected.

Conclusion

We found a great variety of mutations depending on the analyzed region in patients with SNHL; 57.86% of patients had affection in one or two alleles in GJB2 or GJB6 genes whereas 42.14% were wild-type. In some cases, allele distribution depended on region. Molecular studies of more genes involved in hereditary non-syndromic SNHL are required to completely confirm the molecular basis of hearing loss in Mexican population.     

Genetic similarity between adenoid tissue and middle ear fluid isolates of Streptococcus pneumoniae,Haemophilus influenzae and Moraxella catarrhalis from Iranian children with otitis media with effusion

Background

Otitis media with effusion (OME) is a common disease among children, in the pathogenesis of which bacterial infections play a critical role. It was suggested that adenoid tissue could serve as a reservoir for bacterial infection, the eustachian tubes being the migration routes of bacteria into the middle ear cavity. The aim of this study was to investigate the genetic similarity between isolates of Streptococcus pneumoniae, Haemophilus influenzae and Moraxella catarrhalis, obtained from adenoid tissue and middle ear fluid.

Methods

A total of 60 specimens of middle ear fluids (MEFs) and 45 specimens of adenoid tissue were obtained from 45 children with OME. All the samples were inoculated on culture media for bacterial isolation and identification. The genetic similarity between bacterial isolates was determined by pulsed-field gel electrophoresis (PFGE).

Results

The same bacterial species were simultaneously isolated from adenoid tissue and MEFs of 14 patients, among which, 6 pairs of M. catarrhalis, 5 pairs of S. pneumoniae and 3 pairs of H. influenzae were identified.

Conclusions

Based on the genetic similarities between isolate pairs, found by PFGE analysis, this study suggested that M. catarrhalis, S. pneumoniae and H. influenzae colonize the adenoid tissue, then migrate to the middle ear cavity and, hence, contribute to the total pathogenesis of OME.     

Epidemiology of nasopharyngeal colonization by S. pneumoniae in Indian infants in the first 2 years of life

Background

Streptococcus pneumoniae is a leading cause of invasive disease in developing countries like India. Although the 13 valent pneumococcal vaccine has already been introduced in the country, there is very little epidemiological data regarding S. pneumoniae colonization and antibiotic susceptibility in Indian infants.

Methods

We studied serogroup/serotype (SGT) distribution and antibiotic susceptibility pattern of S. pneumoniae in unvaccinated Indian infants by performing monthly nasopharyngeal swabbing of a birth cohort for 2 years.

Results

Colonization began soon after birth and was complete in the first year of life in the majority of those colonized. Carriage rates increased during winter (p < 0.01) and in those with upper respiratory infection (URI) (p < 0.01). The most frequently (76.1%) isolated SGT were 19, 6, 15, 23, 9, 35 and 10. Vaccine SGT accounted for 60.5% of all colonizers. Antibiotic resistance was maximum for cotrimoxazole (94.3%) and least for erythromycin (11.2%) with no penicillin resistance. Ten of the commonest SGT which cause invasive disease among Indian infants comprised 46.9% of the colonizers. Serogroups 1, 5, 45 and 12 which cause invasive disease in under-fives were not seen in this birth cohort in the first year.

Conclusions

S. pneumoniae colonization in Indian infants commences soon after birth and chiefly occurs in the first year of life. The 13 valent vaccine may protect against a little less than half the commonly seen invasive SGT of S. pneumoniae.     

Nasopharyngeal Streptococcus pneumoniae carriage in Japanese children attending day-care centers

Objectives

We conducted a prospective bacteriological survey to investigate antibiotic resistance-related genetic characteristics and the turnover of nasopharyngeal Streptococcus pneumoniae carriage in healthy children in day-care centers (DCCs).

Methods

A total of 363 nasopharyngeal mucus samples were collected from children aged 0 to 6 years attending two DCCs in the summer of 2004 (n = 181) and the following winter (n = 182). We obtained 157 S. pneumoniae isolates and analyzed them by antibiotic susceptibility testing, PCR assay for the penicillin-binding protein (PBP) genes and macrolide-resistance gene, and pulsed-field gel electrophoresis (PFGE).

Results

The overall carriage rate was 43.3% (157/363). The percentages of penicillin-intermediately resistant S. pneumoniae (PISP) strains, penicillin-resistant S. pneumoniae (PRSP) strains, erythromycin-intermediately resistant S. pneumoniae strains and erythromycin-resistant S. pneumoniae strains were 35.7% (56/157), 0.6% (1/157), 1.9% (3/157), and 69.4% (109/157), respectively. The percentages of S. pneumoniae strains with the pbp mutation(s) and mefA and/or ermB gene(s) were 92.4% (145/157) and 71.3% (112/157), respectively. Fifty strains with different PFGE patterns were obtained from among the 157 isolates. Thirteen strains were observed in both seasons, but only one of these strains was isolated from the same carrier. Twenty-one strains (42.0%) were isolated from two or more children, and 17 of these were each isolated from children attending the same DCC.

Conclusions

These results indicate the spread of S. pneumoniae, particularly those with antibiotic-resistance genes, and the vigorous genetic turnover and substantial horizontal transmission of this pathogen in healthy children attending DCCs in Japan.