Changing clinical manifestations of Gaucher disease in Taiwan.

BACKGROUND: Gaucher disease (GD) is a lysosomal storage disorder characterized by deficient glucocerebrosidase activity that results from biallelic mutations in the GBA1 gene. Its phenotypic variability allows GD to be classified into 3 subtypes based on the presence and extent of neurological manifestations. Enzyme replacement therapy (ERT) has been available for all patients with GD in Taiwan since 1998. Newborn screening (NBS) for GD has been available since 2015. This study attempted to unveil the clinical features of patients diagnosed with GD during different eras in Taiwan. MATERIALS AND METHODS: Data from the health records of two tertiary hospitals responsible for two-thirds of the patients with GD in Taiwan were used. The study population included all patients identified as having GD between 1998, and April 2022, in these two hospitals for review. A total of 42 individuals were included, six of whom were diagnosed by NBS. RESULTS: Our cohort presented a higher proportion of GD3 individuals, both by clinical suspicion and by NBS diagnosis, than that reported worldwide. The major subtypes that were recognized following NBS diagnosis were GD2 and GD3. The majority of GD patients carry at least one p.Leu483Pro variant. The 5-year survival rates were 0% for GD2 patients and 100% for patients with other subtypes. Patients diagnosed during the post-NBS era were free of symptoms on initial presentation, except for those with the GD2 subtype. For those diagnosed earlier, ERT was shown to be effective in terms of improved hemograms and prevented bone crises. However, the neurological symptoms in GD3 patients progressed despite ERT intervention. CONCLUSION: ERT is essential in reversing the hematological presentations and preventing the skeletal complications of GD. Timely diagnosis of GD with NBS allows for early intervention with ERT to prevent disease progression and complications. However, the need for effective intervention for neurological dysfunction remains unmet.

Rapid and sensitive detection of carbapenemase activity in Acinetobacter baumannii using superficially porous liquid chromatography-tandem mass spectrometry.

BACKGROUND: The emergence and spread of carbapenem-resistant Acinetobacter baumannii poses a challenge for optimizing antibiotic therapies and preventing outbreaks. Traditional phenotypic assays such as the modified Hodge test (MHT) or polymerase chain reaction (PCR)-based detection of the carbapenemase genes are time-consuming and complicated. Therefore, new approaches for the efficient detection of carbapenemase-producing A. baumannii are urgently required. METHODS: In this study, we used the superficially porous liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay to measure carbapenem hydrolysis in a solution spiked with test strains of A. baumannii. The rate of carbapenem hydrolysis during incubation was expressed as the ratio of the carbapenem peak area of the test A. baumannii strains to the noncarbapenemase-producing A. baumannii ATCC 17978. This method can accurately measure the carbapenem hydrolysis rate and, therefore, can effectively identify carbapenemase-producing strains within 75 minutes. RESULTS: A total of 112 A. baumannii strains were used in this study, including 103 clinical isolates with 68 carbapenem-resistant strains and 35 carbapenem-susceptible strains, seven ATCC strains and two selected mutants. The results of the superficially porous LC-MS/MS assay showed higher detection sensitivity compared to the results of the MHT. CONCLUSION: Our results demonstrate the ability of the former method to routinely detect carbapenemase-producing A. baumannii.

Improved identification of multiple drugs of abuse and relative metabolites in urine samples using liquid chromatography/triple quadrupole mass spectrometry coupled with a library search.

RATIONALE: Although two multiple reaction monitoring (MRM) transitions per compound are used for identification performed using liquid chromatography/triple quadrupole mass spectrometry (LC/QqQ-MS/MS), differences in identification criteria among several regulations may lead to misidentification. We demonstrated that the use of two MRM transitions and product ion spectra improves compound identification. METHODS: The scan cycle time was reduced using time-scheduled MRM (tMRM), data-dependent product ion scanning, and dynamic exclusion. The quantification and identification performance for 13 drugs of abuse and their metabolites were evaluated. RESULTS: Deuterated internal standards compensated for ion suppression. All analytes exhibited intra- and interday precision <12.11%, accuracy of -10.31% to +10.10%, and no carryover. The LC/QqQ-MS/MS and reference gas chromatography/MS methods were equally precise, accurate, and specific. Several regulatory organizations include two MRM transitions, their ratio, and retention time as identification criteria. In 28 samples, the relative ion ratio variation was >10% and product ion spectral matches with >94% probabilities improved drug and metabolite identification. CONCLUSIONS: The LC/QqQ-MS/MS method is a comprehensive assay in which tMRM and the product ion scan are combined in a single run by using a QqQ mass analyzer to simultaneously quantify amphetamine, ketamine, morphine, and their relative metabolites in urine. The proposed method can be applied in forensic science.