SPECT Imaging of P. aeruginosa Infection in Mice Using 123I-BMIPP.

Pseudomonas aeruginosa infection is an infectious disease that must be controlled because it becomes chronic and difficult to treat, owing to its unique system of toxin production/injection and elimination of other bacteria. Here, we noninvasively monitored P. aeruginosa using single-photon emission computed tomography (SPECT) imaging. Determining the amount and localization of the P. aeruginosa will enable making faster clinical diagnoses and selecting the most appropriate therapeutic agents and methods. Nonclinically, this information can be used for imaging in combination with biofilms and toxin probes and will be useful for discovering drugs targeting P. aeruginosa. To study P. aeruginosa accumulation, we conducted in vitro and in vivo studies using iodine-123 ß-methyl-p-iodophenyl-pentadecanoic acid (123I-BMIPP), which we previously reported using for Escherichia coli. In vitro, 123I-BMIPP accumulated in P. aeruginosa by being taken up into the bacteria and adsorbing to the bacterial surface. In vivo, 123I-BMIPP accumulated significantly more in infected sites than in noninfected sites and could be quantified by SPECT. These results suggest that 123I-BMIPP can be used as a probe for P. aeruginosa for SPECT. Establishing a noninvasive monitoring method using SPECT will allow further progress in studying P. aeruginosa.

The error patterns of phonemes in children with prelingual hearing loss: A comparison between hearing aid and cochlear implant users.

OBJECTIVE: To reveal differences in error pattern of phonemes and articulation between children using cochlear implants (CIs) and those using hearing aids (HAs) due to prelingual hearing disorder and help the education of children with prelingual hearing loss. METHOD: Children with prelingual hearing loss who were receiving auditory-verbal preschool education at an auditory center for hearing-impaired children (Fujimidai Auditory Center, Tokyo, Japan) from 2010 to 2020 were analyzed retrospectively. All participants underwent pure tone audiometry and monosyllabic intelligibility tests. The error answers were categorized into five patterns which was characterized by the substitution, addition, omission, failure, and no response according to consonant errors. In addition, the consonant errors classified into the manner of articulation and the differences of error patterns were analyzed between the HA and the CI group descriptively. RESULTS: A total of 43 children with bilateral HAs and 46 children with bimodal CIs or bilateral CIs were enrolled. No significant between-group differences in median phoneme intelligibility were found. The most common error pattern was substitution in both HA and CI groups. The error number of addition pattern in the HA group was smaller than in the CI group. In both groups, the most common errors of articulation were flap errors, and the most common error patterns were flaps to nasals, nasals to nasals, plosives to plosives. In the HA group, plosives and nasals tended not to be recognized and in the CI group plosives were prone to be added to vowels. CONCLUSIONS: There were some different error patterns of articulation and consonant substitution between groups. Clarifying differences of phoneme that are difficult to hear and tend to be misheard would help for creating an effective approach to auditory training for children with hearing loss.

Evaluation of L-Alanine Metabolism in Bacteria and Whole-Body Distribution with Bacterial Infection Model Mice.

The World Health Organization has cautioned that antimicrobial resistance (AMR) will be responsible for an estimated 10 million deaths annually by 2050. To facilitate prompt and accurate diagnosis and treatment of infectious disease, we investigated the potential of amino acids for use as indicators of bacterial growth activity by clarifying which amino acids are taken up by bacteria during the various growth phases. In addition, we examined the amino acid transport mechanisms that are employed by bacteria based on the accumulation of labeled amino acids, Na+ dependence, and inhibitory effects using a specific inhibitor of system A. We found that 3H-L-Ala accurately reflects the proliferative activity of Escherichia coli K-12 and pathogenic EC-14 in vitro. This accumulation in E. coli could be attributed to the amino acid transport systems being different from those found in human tumor cells. Moreover, biological distribution assessed in infection model mice with EC-14 using 3H-L-Ala showed that the ratio of 3H-L-Ala accumulated in infected muscle to that in control muscle was 1.20. By detecting the growth activity of bacteria in the body that occurs during the early stages of infection by nuclear imaging, such detection methods may result in expeditious diagnostic treatments for infectious diseases.

Three cases of otitis media caused by Mycobacterium abscessus subsp. abscessus: Importance of medical treatment and efficacy of surgery.

This study aimed to assess the clinical presentation, antibiotic therapy, surgery, and outcomes in patients with otitis media caused by Mycobacterium abscessus subsp. abscessus and discuss the efficacy of surgery. This is a retrospective case review of three patients diagnosed with otomastoiditis caused by M. abscessus subsp. abscessus. All patients had refractory otorrhea. One patient had granulation tissue in the tympanic membrane. They received medical treatment and underwent surgery. Otorrhea was resolved several months after the initiation of long-term multiantibiotic therapy in all cases. The timing of surgery varied among patients. Before initiating antibiotic therapy, mastoidectomy was performed to achieve definitive diagnosis in two patients, and wound dehiscence developed in these patients. Two patients underwent debridement after the initiation of multiantibiotic therapy. After antibiotic administration, tympanoplasty was performed to improve hearing in one patient. All patients achieved culture negativity after treatment, and no recurrences have been noted. From three cases, it is suggested that the mainstay of treatment for M. abscessus subsp. abscessus is long-term multiantibiotic therapy, and surgery itself may have little effect on achieving ear dryness. Thus, in most patients, drug therapy should be prioritized. Considering postoperative complications, surgery before achieving ear dryness should be avoided, except in emergency cases. In addition, if the diagnosis is not confirmed by repeated bacteriological tests, mastoidectomy should be performed to collect specimens. Tympanoplasty for hearing loss or eardrum perforation is recommended after discontinuation of medications.

Assessment of speech perception in deaf or hard of hearing children who received auditory-verbal therapy with hearing aids or cochlear implants.

OBJECTIVES: To reframe the criteria for pediatric cochlear implants (CIs) in Japan, we investigated monosyllabic and word speech perception at the time of school entry in deaf or hard of hearing (D/HH) children who underwent auditory-verbal therapy (AVT) with hearing aids (HAs) or CIs in early childhood. METHODS: D/HH children who started AVT at Fujimidai Auditory Center for Hearing-Impaired Children before the age of 1 year and who underwent auditory assessment in the previous year of schooling (at the age of 5 or 6) from 2010 to 2020 were enrolled in this study. The results of hearing level tests with or without amplification and monosyllabic and word speech discrimination were assessed. RESULTS: The children with profound hearing impairment (>90 dB HL) had worse speech perception than children with CIs. The children with severe hearing impairment (>70-90 dB HL) did not show any significant differences in speech perception compared with the children with CIs. The children with HAs with a wearing threshold <40 dB HL did not show any significant differences in speech perception from the children with CIs. CONCLUSION: D/HH children with audiologic thresholds greater than 90 dB HL or aided thresholds greater than 40 dB HL should be considered for CIs to ensure development of appropriate listening and spoken language.

A highly sensitive and specific method to evaluate viable fungal burden of Aspergillus fumigatus in mice by RT-qPCR for 18S ribosomal RNA.

Potent fungicidal activity is one of the key factors of antifungals to overcome invasive pulmonary aspergillosis (IPA). To date, quantification of Aspergillus DNA in the lungs and galactomannan (GM) in serum or bronchoalveolar lavage fluid have been developed as general methods for measuring fungal burden in IPA animal models. However, GM quantification is not supposed to be a suitable method for precise evaluation of the fungicidal effects of antifungals, because killed Aspergillus hyphae can release GM for a certain period until they are eliminated by the host. Therefore, in terms of detecting viable fungal burden of Aspergillus, quantification of Aspergillus DNA has been thought to be a suitable method. Here, to obtain a method with much higher sensitivity, we applied reverse transcription quantitative PCR (RT-qPCR) for A. fumigatus 18S ribosomal RNA to measure the viable fungal burden in murine IPA models. Prior to in vivo tests, we confirmed that the sensitivity of 18S rRNA was nearly 50-fold higher than that of 18S ribosomal DNA in vitro. This highly sensitive method made it possible to evaluate the fungicidal effects of antifungals in a low-inoculation murine IPA model. In this model, single administrations of higher doses of voriconazole and posaconazole, which have fungicidal activity, were able to display fungicidal effects with ≥1 log10 reductions by 18S rRNA quantification, whereas significant reductions in serum GM were not observed. These results suggest that 18S rRNA quantification is a powerful tool for screening novel antifungals with potent fungicidal activity only after a single administration.

A case with acquired lacrimal fistula due to Sjögren's syndrome.

PURPOSE: To report a case with lacrimal fistula presumably associated with Sjögren's syndrome. OBSERVATIONS: A patient with Sjögren's syndrome was referred to our hospital for fistula from the inferior lacrimal canaliculus to the palpebral conjunctiva of her left lower eyelid. She also revealed severe dry eye in both eyes. She had no history of trauma or congenital lacrimal fistula. She was administering 0.3% purified sodium hyaluronate and 3% diquafosol sodium solution 6 times a day for dry eye. The Schirmer-1 test indicated that tear secretion was 0 mm for 5 min for both eyes. She was diagnosed with Sjögren's syndrome based on increased levels of blood Sjögren's-syndrome-related antigen A and Sjögren's-syndrome-related antigen B antibodies, decreased saliva volume, and lip biopsy. We performed silicone tube intubation and patched the fistula with conjunctiva. We observed the lacrimal sac and nasolacrimal duct under lacrimal micro-endoscopy; there was no bacterial concretion, obstruction, and inflammation of lacrimal mucosa. After the operation, her symptoms improved and lacrimal perforation healed after the removal of the silicone tube. CONCLUSIONS AND IMPORTANCE: Sjögren's syndrome can cause not only corneal perforation but also mucosal perforation, which may lead to a lacrimal fistula. Sjögren's syndrome patients with severe dry eye should be managed carefully.

Macular Structure Recovery after Surgery for Optic Disc Pit Maculopathy.

The present report aimed to describe the macular structure's recovery process in a case of optic disc pit maculopathy (ODP-M) with outer layer hole following pars plana vitrectomy (PPV) with juxtapapillary laser treatment (JPL). We performed repeated optical coherence tomography (OCT) examinations to evaluate the macular structural changes. An 80-year-old man presented with distorted vision and decreased visual acuity (VA) in his left eye, experienced for 1 year, prior to presentation. Fundus examination and OCT showed intraretinal fluid (IRF) in the inner and outer retinal layers. Serous retinal detachment (SRD) with an outer layer hole in the macula was also evident. The IRF was connected to the optic disc; however, the SRD was isolated. Best-corrected VA was 20/100. PPV combined with JPL was performed. Posterior vitreous detachment creation and tamponade with sulfur hexafluoride was performed. Postoperatively, the inner retinal IRF at the fovea disappeared. The outer layer hole gradually closed and had completely disappeared 1 month postoperatively. After resolution of the outer layer hole, SRD reduced gradually and disappeared 8 months postoperatively, although the macular outer retinal IRF remained. The outer retinal IRF had partially resolved by the 18th postoperative month. Macular structure was completely recovered 31 months postoperatively, with an improved VA of 20/20. In conclusion, SRD might be associated with outer retinal IRF and outer layer holes. In cases of ODP-M, outer layer holes might induce optic disc-isolated SRD.

Generation, characterization and therapeutic potential of anti-feline TNF-alpha MAbs for feline infectious peritonitis.

Feline infectious peritonitis (FIP) is a lethal infectious disease affecting domestic and wild cats. Several reports suggested that TNF-alpha is related to the progression of FIP. Thus, the administration of a feline TNF-alpha-neutralizing antibody to cats with FIP may reduce the disease progression. In this study, we have prepared nine monoclonal antibodies (MAbs) that recognize feline TNF-alpha. All MAbs neutralized recombinant TNF-alpha. The 50% inhibitory concentrations (IC50) of the MAbs for the cytotoxicity of recombinant TNF-alpha were 5-684 ng/ml. MAb 2-4 exhibited high neutralizing activity against natural TNF-alpha derived from FIPV-infected macrophages, and was confirmed to inhibit the following feline TNF-alpha-induced conditions in vitro: (i) an increase in the survival rate of neutrophils from cats with FIP, (ii) aminopeptidase N (APN) mRNA expression in macrophages, and (iii) apoptosis of a feline T-lymphocyte cell line.

Defective glycosylation of α-dystroglycan contributes to podocyte flattening.

In addition to skeletal muscle and the nervous system, α-dystroglycan is found in the podocyte basal membrane, stabilizing these cells on the glomerular basement membrane. Fukutin, named after the gene responsible for Fukuyama-type congenital muscular dystrophy, is a putative glycosyltransferase required for the post-translational modification of α-dystroglycan. Chimeric mice targeted for both alleles of fukutin develop severe muscular dystrophy; however, these mice do not have proteinuria. Despite the lack of a functional renal defect, we evaluated glomerular structure and found minor abnormalities in the chimeric mice by light microscopy. Electron microscopy revealed flattening of podocyte foot processes, the number of which was significantly lower in the chimeric compared to wild-type mice. A monoclonal antibody against the laminin-binding carbohydrate residues of α-dystroglycan did not detect α-dystroglycan glycosylation in the glomeruli by immunoblotting or immunohistochemistry. In contrast, expression of the core α-dystroglycan protein was preserved. There was no statistical difference in dystroglycan mRNA expression or in the amount of nephrin and α3-integrin protein in the chimeric compared to the wild-type mice as judged by immunohistochemistry and real-time RT-PCR. Thus, our results indicate that appropriate glycosylation of α-dystroglycan has an important role in the maintenance of podocyte architecture.