Yusuke Minato

OBJECTIVE: Mycobacterium lentiflavum is a rare, non-tuberculous mycobacterium (NTM) which is implicated in some cases of active, pulmonary non-tuberculous mycobacterial disease. The outbreak of NTM in nosocomial settings occasionally occurs and outbreak investigation with implementation of concurrent countermeasure is essential. DESIGN: Outbreak investigation. SETTING: A tertiary care medical center. PATIENTS AND PARTICIPANTS: Hospitalized patients during the outbreak period. RESULTS: In April 2024, a cluster of patients with Mycobacterium lentiflavum-positive sputum cultures, presumed to be due to nosocomial transmission, was identified at the study center. A retrospective review of cases dating back to February 2023 revealed 27 patients with M. lentiflavum infection whose isolates had initially not been speciated. According to the American Thoracic Society (ATS) criteria for diagnosing pulmonary non-tuberculous mycobacterial (NTM) disease, two of these patients met the criteria for active disease. Multi-locus sequence typing of 12 isolates demonstrated 100% clonality, indicating a common source. A concurrent outbreak investigation identified contaminated faucet aerators in hospital wards as the likely source of transmission. All faucet aerators and caps were manually cleaned and disinfected using liquid sodium hypochlorite, after which no further cases were detected. CONCLUSION: The present study described a nosocomial cluster of M. lentiflavum colonization and infections at a tertiary care center, with contaminated faucet aerators identified as the likely source. Prompt identification of such NTM clusters in healthcare settings is essential to initiate timely treatment and prevent further transmission.

Mycobacterium intracellulare is a major etiological agent of the recently expanding Mycobacterium avium-intracellulare complex pulmonary disease (MAC-PD). Therapeutic regimens that include a combination of macrolides and antituberculous drugs have been implemented with limited success. To identify novel targets for drug development that accommodate the genomic diversity of M. avium-intracellulare, we subjected eight clinical MAC-PD isolates and the type strain ATCC13950 to genome-wide profiling to comprehensively identify universally essential functions by transposon sequencing (TnSeq). Among these strains, we identified 131 shared essential or growth-defect-associated genes by TnSeq. Unlike the type strain, the clinical strains showed increased requirements for genes involved in gluconeogenesis and the type VII secretion system under standard growth conditions, the same genes required for hypoxic pellicle-type biofilm formation in ATCC13950. Consistent with the central role of hypoxia in the evolution of M. intracellulare, the clinical MAC-PD strains showed more rapid adaptation to hypoxic growth than the type strain. Importantly, the increased requirements of hypoxic fitness genes were confirmed in a mouse lung infection model. These findings confirm the concordant genetic requirements under hypoxic conditions in vitro and hypoxia-related conditions in vivo and highlight the importance of using clinical strains and host-relevant growth conditions to identify high-value targets for drug development.

BACKGROUND: Flomoxef and cefmetazole have been reported to be effective against broad-spectrum β-lactamase-producing bacteria and have gained attention as a potential alternative to carbapenems. This study aimed to compare the efficacy of these two drugs in treating urinary tract infection (UTI) by integrating in vitro data and two real-world databases. METHODS: The susceptibility rates of third-generation cephalosporin-resistant Escherichia coli and Klebsiella pneumoniae to flomoxef and cefmetazole were compared using comprehensive national antimicrobial resistance surveillance data. Combined antimicrobial activities against an extended-spectrum beta-lactamase (ESBL)-producing multidrug-resistant bacterial strain were tested by diagonal measurement of n-way drug interactions. The effectiveness of the two drugs in treating UTIs was compared using hospital stay duration data obtained from the Japan Medical Data Center (JMDC) Claims Database. RESULTS: Third-generation cephalosporin-resistant E. coli and K. pneumoniae, including ESBL-producing strains, were similarly susceptible to flomoxef and cefmetazole. In vitro assessment against an ESBL-producing multidrug-resistant strain revealed similar antimicrobial interaction patterns. JMDC Claims data analysis showed that the median hospital stay was 11 (95% confidence interval [CI]: 11-11) and 4 (95% CI: 3-5) days in the cefmetazole and flomoxef groups, respectively (log-rank test, P < 0.001). Moreover, the flomoxef group demonstrated a significantly lower frequency of adverse events such as C. difficile infection and renal failure. CONCLUSIONS: The effectiveness of flomoxef is comparable to that of cefmetazole in treating UTIs without major complications. Thus, flomoxef is a viable treatment option for UTIs in locales with a high prevalence of ESBL-producing strains.

OBJECTIVE: To examine the international trends for nontuberculous mycobacterial-associated mortality rates, as nontuberculous mycobacterial infections are becoming increasingly prevalent and pose a significant public health challenge, especially in older populations. METHODS: This retrospective observational study used data from the World Health Organization mortality database, which included patients with nontuberculous mycobacterial infection in 83 countries. We stratified the data by sex, age, and geographic region and calculated crude and age-standardized mortality rates to estimate long-term mortality trends. RESULTS: In total, 42,182 nontuberculous mycobacterial infection-associated deaths (58.1% in women) were reported in 83 countries between 2000 and 2022. The locally weighted regression model estimation for the nontuberculous mycobacterial infection-associated mortality rate more than doubled-from 0.36 deaths per 1,000,000 individuals in 2000 to 0.77 deaths per 1,000,000 individuals in 2022. Eighty-six percent of nontuberculous mycobacterial infection-associated deaths occurred in people aged ≥65 years. The mortality rate was the highest in the Western Pacific Region. CONCLUSIONS: This study highlights the impact of emerging nontuberculous mycobacterial diseases and the importance of targeted interventions for managing and reducing mortality, particularly in vulnerable older populations. Further studies are warranted to determine the factors contributing to geographical disparity and treatment options.

This study highlights the novel potential of molecular aggregates as inhibitors of a disease-related protein. Enzyme inhibitors have been studied and developed as molecularly targeted drugs and have been applied for cancer, autoimmune diseases, and infections. In many cases, enzyme inhibitors that are used for therapeutic applications interact directly with enzymes in a molecule-to-molecule manner. We found that the aggregates of a small compound, Mn007, inhibited bovine pancreatic DNase I. Once Mn007 molecules formed aggregates, they exhibited inhibitory effects specific to DNases that require divalent metal ions. A DNase secreted by Streptococcus pyogenes causes streptococcal toxic shock syndrome (STSS). STSS is a severe infectious disease with a fatality rate exceeding 30% in patients, even in this century. S. pyogenes disrupts the human barrier system against microbial infections through the secreted DNase. Until now, the discovery/development of a DNase inhibitor has been challenging. Mn007 aggregates were found to inhibit the DNase secreted by S. pyogenes, which led to the successful suppression of S. pyogenes growth in human whole blood. To date, molecular aggregation has been outside the scope of drug discovery. The present study suggests that molecular aggregation is a vast area to be explored for drug discovery and development because aggregates of small-molecule compounds can inhibit disease-related enzymes.