Longitudinal changes in dynamic characteristics of neonatal external and middle ears.

OBJECTIVES: Neonates have smaller and less mature ears than adults. Developmental changes in structure and function continually occur after birth and may affect the diagnostic results obtained by audiometric assessment instrumentation, such as tympanometry and otoacoustic emission. In the present study, we investigated longitudinal changes in external and middle ear dynamic characteristics by performing sweep frequency impedance (SFI) tests. METHODS: SFI tests were longitudinally performed on healthy Japanese neonates (1 female and 1 male) from birth to 3 and 5 months, respectively. A sound of sweeping sinusoidal frequency, ranging from 0.1 kHz to 2 kHz, was presented to the ear canal at 50-daPa intervals of static pressure from +200 to -200 daPa. Test results were expressed a curve showing the sound pressure level (SPL) relative to probe tone frequency, called SPL curve. RESULTS: The first fluctuation in resonance frequency (RF1) and SPL (ΔSPL1), related to the external ear, showed significant developmental changes as chronological age increased; that is, RF1 and ΔSPL1 were respectively increased and decreased and thereafter became unmeasurable by 5 months of age. In contrast, the second fluctuation in resonance frequency (RF2) and SPL (ΔSPL2), related to the middle ear, did not show significant changes over the measurement period. CONCLUSIONS: The present results suggest that the dynamic characteristics of the external ear canal wall changed with increases in chronological age; the resonance of the wall at about 0.3 kHz at birth tended to increase to about 0.7 kHz and to be unmeasurable by 5 months of age, while those of the middle ear did not significantly changed. These results showing how neonatal-ear dynamics changes with chronological age may be an important key in further hearing research and the development of hearing devices and diagnostic tools suitable for neonates.

The expression of 11 beta hydroxysteroid dehydrogenase in severe allergic rhinitis.

OBJECTIVE: To clarify the roles of 11 beta-HSD in resistance to glucocorticoid therapy for allergic rhinitis, a case series study was conducted. METHODS: The patient group consisted of 20 subjects with allergic rhinitis, aged from 21 to 46 years (mean age 26.5), who showed persistent GC resistance necessitating surgical removal of the inferior turbinate after 6 months' GC treatment. The patients with poor response to GC treatment for 6 months' were defined as GC resistance. The control group consisted of 10 subjects aged from 16 to 39 years (mean age 24.5) who underwent maxillofacial surgery, from whom nasal tissues were taken and who did not receive GC treatment. Nasal mucosal tissues from patients and cntorol subjects were examined immunohistochemically. The sections were washed with 0.01 M phosphate-buffered saline (PBS; pH 7.2) containing 0.15 M NaCl and 0.01% Triton X-100, and incubated for 2 h with rabbit polyclonal anti-11 beta HSD1 and 11 beta-HSD2 antibody (Santa Cruz Biotechnology, Inc., Santa Cruz, CA, USA), each diluted 1:200 in PBS containing 0.1% bovine serum albumin. Immunostained sections were assessed under an Olympus microscope with an eyepiece reticule at 200 X magnification. Cell counts are expressed as means per high-power field (0.202 mm2). Control group means (arithmetic mean ± SD) were compared with patient group means by Mann-Whitney U-test at P = 0.05. RESULTS: Although 11 beta-HSD1 was expressed to a similar extent in patients and controls, 11 beta-HSD2 was expressed significantly more in patients with severe allergic rhinitis, resulting in a increased HSD-1/HSD-2 ratio. The significantly increased expression of 11 beta-HSD2 in the nasal epithelium and submucosal inflammatory cells of patients with severe nasal allergy were observed in the present study. CONCLUSION: Our findings suggest that 11 beta-HSD2 plays an important role in resistance to glucocorticoid therapy for allergic rhinitis, and its expression might be used as an additional parameter indicating steroid resistance in allergic rhinitis.

Possible interaction between the bacterial transcription factor ArtA and the eukaryotic RNA polymerase III promoter.

Eukaryotic RNA polymerase III (RNAP III) transcribes tRNA genes and short interspersed elements that have internal promoters consisting of A- and B-blocks. The B-block binding subunit of the transcription initiation factor TFIIIC binds to the B-block. The mobile bacterial insertion sequence (IS) 1 contains a RNAP III promoter-like sequence, which stimulates bacterial transcription along with the bacterial ArtA protein. Here, the DNA-binding ability of ArtA was examined in vitro using a simple, newly developed method. Various DNA fragments, including RNAP III promoter fragments, were separately incubated with purified ArtA, and then loaded onto a polyacrylamide gel. Since DNAs bound by ArtA remain in the gel wells during electrophoresis, SDS was added into the wells at the electrophoresis halfway point. It was hypothesized that SDS would dissociate the DNA-ArtA complexes in the wells, and then the DNAs would begin to migrate. In fact, new bands appeared in all of the lanes at similar intensities, indicating that ArtA binds nonspecifically to DNA. Therefore, labeled wild-type RNAP III promoter fragments were incubated with either the unlabeled wild-type or mutant fragments and ArtA, and electrophoresed. The B-block(-like) sequences of IS1, a human Alu element, and an anuran tRNA gene were important for binding to ArtA. Additionally, in silico analyses revealed the presence of the RNAP III promoter-like structures in the IS1 isoforms and the IS3 family elements. These results suggest the presence of parts of the RNAP III transcription machinery in bacteria, and might imply that its prototype existed in the common ancestor.

Evolution of the B-Block Binding Subunit of TFIIIC That Binds to the Internal Promoter for RNA Polymerase III.

Eukaryotic RNA polymerase III transcribes tRNA genes, and this requires the transcription factor TFIIIC. Promoters are within genes, with which the B-block binding subunit of TFIIIC associates to initiate transcription. The binding subunits are more than 1000 amino acids in length in various eukaryotic species. There are four regions with conserved sequence similarities in the subunits. The helix-turn-helix motif is included in one of these regions and has been characterized as the B-block_TFIIIC family in the Pfam database. In the NCBI and EMBL translated protein databases, there are archaeal proteins (approximately 100 amino acids in length) referred to as B-block binding subunits. Most of them contain a B-block_TFIIIC motif. DELTA-BLAST searches using these archaeal proteins as queries showed significant multiple blast hits for many eukaryotic B-block binding subunits on the same proteins. This result suggests that eukaryotic B-block binding subunits were constituted by repeating a small unit of B-block_TFIIIC over a long evolutionary period. Bacterial proteins have also been annotated as B-block binding subunits in the databases. Here, some of them were confirmed to have significant similarities to B-block_TFIIIC. These results may imply that part of the RNAP III transcription machinery existed in the common ancestry of prokaryotes and eukaryotes.

Unusual parapharyngeal extension of peritonsillar abscess to the masticator space: successfully drained by extraoral and intraoral endoscopic approaches.

OBJECTIVE: The object was to describe 2 novel cases of peritonsillar abscess showing peculiar extension to the masticator space. METHODS: The methods included clinical case records, including computed tomography and surgical approaches. RESULTS: Both patients we encountered were suffering from systematic diseases, with case 1 involving a 75-year-old man with diabetes mellitus and case 2 involving a 90-year-old woman taking immunosuppressive medications. The abscesses were peritonsillar in origin, extending primarily to the parapharyngeal space, with unusual secondary extension to the masticator space. Extraoral drainage conducted in case 1 was useful for assessing the masticator space and surrounding spaces, but endoscopy-assisted intraoral drainage in case 2 was less invasive, obviating the need for identifying the facial nerve. CONCLUSIONS: It is important to bear in mind that patients suffering from systemic diseases may display unusual extension of deep head and neck infections, and enhanced computed tomography is a useful modality for evaluating such extensions.

Dynamic characteristics of the middle ear in neonates.

OBJECTIVE: Early diagnosis and treatment of hearing disorders in neonates is highly effective for realization of linguistic competence and intellectual development. To objectively and quickly evaluate the dynamic characteristics of the middle ear, a sweep frequency impedance (SFI) meter was developed, which allowed the diagnosis of middle-ear dysfunctions in adults and children. However, this SFI meter was not applicable to neonates since the size of the measurement probe was too large. In the present study, therefore, the SFI meter was improved, i.e., the diameter of the probe was reduced to that of the neonatal external ear canal. By using this newly designed SFI meter, SFI tests were performed in healthy neonates. METHODS: A sound of the sweeping sinusoidal frequency between 0.1 kHz and 2.0 kHz in 0.02-kHz step intervals is presented to the ear canal by an SFI probe while the static pressure of the ear canal is kept constant. During this procedure, the sound pressure level (SPL) is measured. The measurements are performed at 50-daPa intervals of static pressure from 200 daPa to -200 daPa. RESULTS: Measurements were conducted in 10 ears of 9 neonates. The SPL showed two variations at 0.26 ± 0.03 kHz and 1.13 ± 0.12 kHz. Since the SPL is known to show a variation at frequencies from 1.0 kHz to 1.6 kHz due to the resonance of the middle ear in adults and children with normal hearing, the second variation is probably related to such resonance in neonates. The measurement of gel models, which mimics the neonatal external ear canal, showed a variation in SPL at around 0.5 kHz. This implies that the source of the first variation may possibly be related to the resonance of the external ear canal wall. CONCLUSIONS: SFI tests revealed that there were two variations in the SPL curve in neonates, one at 0.26 ± 0.03 kHz and the other at 1.13 ± 0.12 kHz, the former and the latter being possibly related to the resonance of the external ear canal wall and that of the middle ear, respectively. This result suggests that the dynamic characteristics of the middle ear in neonates are different from those in adults.

Impact of periodontal treatment in combination with tonsillectomy plus methylprednisolone pulse therapy and angiotensin blockade for pediatric IgA nephropathy.

BACKGROUND: We previously reported the efficacy of extensive eradication of infectious foci in oral and ENT lesions, combined with tonsillectomy plus methylprednisolone (MP) pulse therapy, for curing pediatric Henoch-Schönlein purpura (HSP) and HSP nephritis. In the present study, we used this therapy in patients with pediatric IgA nephropathy (IgAN) to assess whether similar results could be obtained. PATIENTS AND METHODS: In 11 pediatric patients newly diagnosed with IgAN, exploration for infectious foci showed severe oral infection, including dental caries and apical periodontitis, in many. The overall decayed, missing and filled teeth score was elevated to 5.91. Two patients had rhinosinusitis. After extensive treatment of infectious foci, patients underwent tonsillectomy plus MP pulse therapy with angiotensin II receptor blockade. RESULTS: Clinical remission was achieved in all patients with pediatric IgAN (various histologic grades). Remission was achieved by 7.2 ± 5.7 months after initiation of steroid therapy, and disappearance of proteinuria by 3.3 ± 3.0 months. The mean duration of oral steroid administration was 9.5 ± 3.6 months. No relapse has occurred during follow-up of 4.3 ± 2.4 y. CONCLUSIONS: Careful examination and thorough elimination of infectious foci in oral and ENT lesions can optimize the effect of tonsillectomy plus MP pulse therapy, promoting recovery from IgAN.

Periodontal and ENT therapy in the treatment of pediatric Henoch-Schönlein purpura and IgA nephropathy.

Henoch-Schönlein purpura (HSP) and IgA nephropathy (IgAN) are both IgA1-related vasculitis caused by vascular deposition of IgA1-containing immune complexes. A pathological role of the tonsils in the development of HSP and IgAN has been suggested. Tonsils are a mucosaassociated lymphoid organ. Since oral and sinonasal cavities are anatomically directly connected to the tonsils, delivering exogenous antigens into the tonsils to induce local and systemic antibody responses, we examined the infectious status of these cavities when we treated HSP and IgAN. In 40 HSP children (6.7±2.5 years), apical periodontitis, rhinosinusitis, and otitis media were identified in 21 (53%, 4.9±2.8 affected teeth), 19 (48%), and 4 (10%) of them, whereas in 11 IgAN children (10.4±2.5 years), these diseases were observed in 6 (55%, 5.8±4.6 affected teeth), 2 (18%), and 0 (0%) of them, respectively. We first treated the patients with extensive eradiation of infectious foci, including antimicrobial treatment and root canal therapy. In 31 HSP patients, such dental and/or ENT therapy resulted in a complete cure without development of nephritis or recurrent attacks. For the remaining 9 HSP and 11 IgAN patients, we further performed tonsillectomy plus methylprednisolone (MP) pulse therapy to control their intractable symptoms, including aggravated purpura, recurrent HSP attacks or nephritis. Using this therapeutic strategy, all of the HSP patients attained clinical remission. All of the IgAN patients with various histological grades also achieved normalization of urinalysis by 7.2±5.7 months after the start of steroid therapy. No relapses were observed in both diseases during followup for 2-10 years. In pediatric HSP and IgAN, apical periodontitis and rhinosinusitis may be involved in abnormal immune responses in both the tonsils and whole body. We conclude that extensive elimination of these infectious foci is beneficial to optimize the effect of tonsillectomy plus MP pulse therapy.

Diagnostic criteria of eosinophilic otitis media, a newly recognized middle ear disease.

OBJECTIVE: Eosinophilic otitis media (EOM) is a newly recognized intractable middle ear disease, characterised by the accumulation of eosinophils in middle ear effusion and middle ear mucosa. Since EOM patients show gradual or sudden deterioration of hearing, it is important to properly diagnose EOM and to start adequate treatment for EOM. We aimed to investigate the clinical risk factors of EOM and to establish the diagnostic criteria of EOM. PATIENTS AND METHODS: We reviewed 138 patients with EOM and 134 age-matched patients with the common type of otitis media with effusion or chronic otitis media as controls. We analyzed the incidence of the following clinical variables in both groups: bilaterality of otitis media, viscosity of middle ear effusion, formation of granulation tissue in the middle ear, response to the treatment for otitis media, deterioration of bone conduction hearing level, and association with other diseases such as bronchial asthma, chronic rhinosinusitis, nasal polyposis, and allergic rhinitis. RESULTS: A high odds ratio was obtained from an association with bronchial asthma (584.5), resistance to conventional treatment for otitis media (232.2), viscous middle ear effusion (201.6), association with nasal polyposis (42.17), association with chronic rhinosinusitis (26.49), bilaterality (12.93), and granulation tissue formation (12.62). The percentage of patients with EOM who were positive for two or more among the highest four items was 98.55%. CONCLUSION: A patient who shows otitis media with effusion or chronic otitis media with eosinophil-dominant effusion (major criterion) and with two or more among the highest four items (minor criteria), can be diagnosed as having EOM. Patients with ear symptoms should have the proper diagnosis of EOM using the proposed diagnostic criteria, and then can receive adequate treatment, resulting in prevention of deterioration of hearing and quality of life.

Efficacy of early dental and ENT therapy in preventing nephropathy in pediatric Henoch-Schönlein purpura.

In a previous study, we demonstrated the benefit of tonsillectomy for early recovery from Henoch-Schönlein purpura (HSP) nephritis (HSPN), suggesting the pathological role of tonsils in HSP (Inoue et al., Clin Nephrol 67:298-305, 2007). In this study, we evaluated the efficacy of extensive eradication of infectious foci directly connected to the tonsils, including those involved in oral as well as ear, nose, and throat (ENT) diseases, in reducing the nephropathy in HSP. For this purpose, we examined the focal points of infection in 40 newly diagnosed HSP patients. After these focal points of infection had been identified, they were extensively eradicated; when the clinical course was intractable, we also considered tonsillectomy. After administering such therapy to HSP patients, we prospectively followed them up for 0.6 to 8 years. The identified focal infections included dental caries in 28 (70%), apical periodontitis in 21 (53%), rhinosinusitis in 19 (48%), tonsillitis in five (13%), and otitis media in four (10%) of the 40 patients. Seventeen patients (43%) had more than two simultaneous infectious foci, whereas, in five (13%), no infectious focus was found. In 32 patients, antimicrobial treatment with concurrent dental and/or ENT therapy resulted in a complete cure without development of HSPN or recurrent attacks. In eight patients, we performed tonsillectomy-adenotonsillectomy to treat their clinical symptoms, including aggravated purpura and recurrent attacks of HSP or HSPN. All patients were completely cured. The overall incidence of HSPN was only three out of the 40 patients (8%). Oral and ENT diseases were found with high percentages in HSP patients. Early and extensive treatment for these lesions and tonsillectomy-adenotonsillectomy for intractable cases may prevent the complication of HSPN, contributing to the early curing of HSP.

Possible presence and role of the promoter sequence for eukaryotic RNA polymerase III in bacteria.

The bacterial repetitive sequence IS1, is a translocatable DNA segment. The internal region of IS1 acts as a cis-element to stimulate RNA synthesis from the upstream promoter. The product of the bacterial artA gene works with this cis-element to stimulate transcription. Eukaryotic genes for small RNAs and short interspersed repetitive elements (SINEs) have internal promoters, transcribed by RNA polymerase III (RNAP III). RNAP III requires the multisubunit protein factor TFIIIC in transcription initiation. TFIIIC contains the B-block binding subunit which recognizes the internal promoter. Here, I report that the eukaryotic RNAP III promoter-like sequence was found in the cis-element of bacterial IS1. Mutations in the cis-element which affect transcription were present in the RNAP III promoter-like sequence. The RNAP III promoter sequence of Alu, which is a human SINE, was cloned into Escherichia coli, and was shown to stimulate bacterial transcription like the cis-element of IS1. Furthermore, the primary structures of ArtA protein and B-block binding subunits were compared. The amino acid sequence of ArtA appeared to be similar to the N- and C-terminal regions conserved in many B-block binding subunits. Prokaryotes and eukaryotes have been thought to have inherent transcription machineries. The results shown here, however, suggest a new aspect of the evolution of the RNAP III transcription machinery.

[Treatment of Henoch-Schönlein purpura nephritis in children with tonsillectomy].

Henoch-Schönlein purpura nephritis (HSPN) is an important complication of Henoch-Sch6nlein purpura (HSP). Pathological findings in the kidney are similar to those for IgA nephritis, which is characterized by the deposition of IgA immune complex in the glomerular mesangium. Since a tonsillectomy is useful for IgA nephritis, this procedure holds great promise for the treatment HSPN. In the present study, we assessed the effectiveness of a tonsillectomy in children with HSPN whose conditions could not be controlled by medication, including predonisone and cyclophosphamide. Seven patients (four boys and three girls) with histologically well-defined HSPN who had undergone a tonsillectomy between 1998 and 2000 and who had been followed for more than 6 postoperative months were retrospectively examined. The age of the patients ranged between 3 and 13 years (average, 7.6 +/- 3.2 years) at the time of operation. Postoperative changes in urinary data were assessed according in the severity of the pathological findings in the kidney and the patient's clinical condition. The severity of the pathological findings in the kidney was determined based on the classification of the International Study of Kidney Disease (ISKDC) and the Index of Glomeruler Lesion (IGL). All patients examined in the present study had an ISKDC classification of over grade II. One patient had a grade IV classification, 3 had a grade III classification, and 3 had a grade II classification. The patients were placed in one of five clinical groups: (1) nephritic-nephrotic syndrome, (2) acute nephritic syndrome, (3) nephritic syndrome, (4) over 1 g/day of proteinuria without hypoalbuminemia or oedema, or 5) below 1 g/day of proteinuria with or without hematuria. One patient was classified in group 1, 1 was group classified in 2, 2 were classified in group 3, 2 were classified in group 4 and 1 was classified in group 5. The mean observation period was 74 +/- 6.4 months. The hematuria and proteinuria resolved in all patients, regardless of their preoperative pathological or clinical severity, within 12 postoperative months. During the observation period, no relapse was observed. Moreover, all medication, including steroid use, was stopped within the observation period. Therefore, a tonsillectomy was considered to be effective for the treatment of children with HSPN whose conditions cannot be controlled using medication.

Links between repeated sequences.

L1 and Alu elements are long and short interspersed retrotransposable elements (LINEs and SINEs) in humans, respectively. Proteins encoded in the autonomous L1 mediate retrotransposition of the nonautonomous Alu and cellular mRNAs. Alu is the only active SINE in the human genome and is derived from 7SL RNA of signal recognition particle. In the other eukaryotic genomes, various tRNA- and 5S rRNA-derived SINEs are found. Some of the tRNA- and 5S rRNA-derived SINEs have partner LINEs of which 3' sequences are similar to those of the SINEs. One of the tRNA-derived SINEs is shown to be mobilized by its partner LINE. Many copies of tRNA and 5S rRNA pseudogenes are present in the human genome. These pseudogenes may have been generated via the retrotransposition process using L1 proteins. Although there are no sequence similarities between L1 and Alu, L1 functionally links with Alu and even cellular genes, impacting on our genome shaping.

Similarities in transcription factor IIIC subunits that bind to the posterior regions of internal promoters for RNA polymerase III.

BACKGROUND: In eukaryotes, RNA polymerase III (RNAP III) transcribes the genes for small RNAs like tRNAs, 5S rRNA, and several viral RNAs, and short interspersed repetitive elements (SINEs). The genes for these RNAs and SINEs have internal promoters that consist of two regions. These two regions are called the A and B blocks. The multisubunit transcription factor TFIIIC is required for transcription initiation of RNAP III; in transcription of tRNAs, the B-block binding subunit of TFIIIC recognizes a promoter. Although internal promoter sequences are conserved in eukaryotes, no evidence of homology between the B-block binding subunits of vertebrates and yeasts has been reported previously. RESULTS: Here, I reported the results of PSI-BLAST searches using the B-block binding subunits of human and Shizosacchromyces pombe as queries, showing that the same Arabidopsis proteins were hit with low E-values in both searches. Comparison of the convergent iterative alignments obtained by these PSI-BLAST searches revealed that the vertebrate, yeast, and Arabidopsis proteins have similarities in their N-terminal one-third regions. In these regions, there were three domains with conserved sequence similarities, one located in the N-terminal end region. The N-terminal end region of the B-block binding subunit of Saccharomyces cerevisiae is tentatively identified as a HMG box, which is the DNA binding motif. Although I compared the alignment of the N-terminal end regions of the B-block binding subunits, and their homologs, with that of the HMG boxes, it is not clear whether they are related. CONCLUSION: Molecular phylogenetic analyses using the small subunit rRNA and ubiquitous proteins like actin and alpha-tubulin, show that fungi are more closely related to animals than either is to plants. Interestingly, the results obtained in this study show that, with respect to the B-block binding subunits of TFIIICs, animals appear to be evolutionarily closer to plants than to fungi.