Empiema pleural por Parvimonas micra en un paciente inmunocompetente: reporte de un caso / Pleural empyema by Parvimonas micra in an immunocompetent patient: a case report

Se presenta el caso de un paciente joven inmunocompetente, con antecedente de tuberculosis pulmonar, que acude al hospital por un cuadro clínico subagudo de fiebre persistente, baja de peso, disnea y abolición del murmullo vesicular. La tomografía de tórax mostró un extenso empiema en hemitórax izquierdo. Se le toman muestras para detección de gérmenes comunes y se le colocan un tubo de drenaje torácico y se inicia antibioticoterapia. La prueba de MALDI-TOF MS identificó a Parvimonas micra, una bacteria anaerobia, comensal de la flora oral, asociado a periodontitis severa, escasamente reportado en empiema pleural, especialmente, en personas inmunocompetentes. En la evaluación odontológica se realizó el diagnóstico de gingivitis y pericoronaritis de la tercera molar. El paciente evolucionó favorablemente. Se sugiere que, en casos de empiemas pleurales subagudos o crónicos, se debe considerar, además de las micobacterias, como agente etiológico al Parvimonas micra, y optar por exámenes como MALDI-TOF MS o secuenciamiento del 16S rRNA, colocación de tubo de tórax, cobertura antibiótica empírica y evaluación odontológica.

We present the case of a young immunocompetent patient, with a history of pulmonary tuberculosis, who attended the hospital with a subacute clinical picture of persistent fever, weight loss, dyspnea and abolition of vesicular murmur. Chest CT scan showed an extensive empyema in the left hemithorax. Samples were taken for detection of common germs. Then, a chest drainage tube was placed and antibiotic therapy started. The MALDI-TOF MS test identified Parvimonas micra, an anaerobic bacterium, commensal to the oral flora, associated with severe periodontitis, but rarely reported in cases of pleural empyema, especially in immunocompetent patients. Gingivitis and pericoronaritis of the third molar were diagnosed during oral evaluation. The patient progressed favorably. Parvimonas micra should be considered as a possible etiological agent in cases of subacute or chronic pleural empyema, in addition to mycobacteria. Tests such as MALDI-TOF MS or 16S rRNA sequencing, chest tube placement, empirical antibiotic coverage and an adequate oral evaluation should be considered in these cases.

Aggregatibacter actinomycetemcomitans Endocarditis in a Patient with Chronic Periodontitis: A Case Report

@#The HACEK organisms consist of the non-influenzae Haemophilus sp., Aggregatibacter sp., Cardiobacterium sp., Eikenella corrodens and Kingella sp. are responsible for a sizable percentage of infective endocarditis cases worldwide with the mortality rate of 18%. Amongst them, Aggregatibacter actinomycetemcomitans is the most common pathogen strongly associated with infective endocarditis. A. actinomycetemcomitans forms part of the oral microbiota and is also the etiological agent of periodontitis. Here, we present a case of a 37-year-old man with underlying obstructive uropathy, that sought treatment for postural hypotension and symptomatic anaemia with fever. Later, he had developed decompensated congestive cardiac failure with aortic regurgitation. A cardiac echocardiogram revealed the presence of vegetation on the aortic valve. Blood culture grew A. actinomycetemcomitans, and he was treated with furosemide and ceftriaxone. A further dental examination showed the patient is having chronic periodontitis, which could be the possible source of A. actinomycetemcomitans causing infective endocarditis. The patient was then transferred to the National Heart Centre for the first time for further management after completion of 4 weeks of intravenous antibiotics. As the pathogen is fastidious, rapid and newer technology like MALDI-TOF mass spectrometry provides rapid and accurate identification for appropriate patient clinical management.

Application of Matrix-assisted Laser Desorption Ionization Mass Spectrometry in Analysis of Traditional Chinese Medicine：A Comprehensive Review / 中国实验方剂学杂志

Traditional Chinese medicine （TCM）， which owns abundant chemical components and complex action pathways， has been widely recognized in the prevention and treatment of diseases. Some analysis methods have been emerged in order to ensure the quality of TCM and to develop new TCM drugs. Matrix-assisted laser desorption ionization mass spectrometry （MALDI-MS） is a soft ionization mass spectrometric technique with the advantages of high throughput， high sensitivity， low cost and so on. It provides technical support for the molecular level study on TCM. At present， this technique has been used in the field of composition analysis and metabonomics research of TCM， and plays an important role in the identification of Chinese herbal medicines， real-time molecular screening and the construction of metabolic network pathway of active ingredients. Among them， the selection of appropriate matrix and sample preparation technology is the key to ensure the detection effect of MALDI-MS. With the development and optimization of new matrix， the continuous improvement of sample preparation technology and the combination of MALDI-MS with various analytical methods will greatly improve the detection effect. Based on this， this paper discusses the application of MALDI-MS in TCM， including high-throughput detection of active ingredients in TCM， monitoring of the original medicines and their metabolites in vivo， and in situ visualization and characterization of tissue distribution information of active ingredients in TCM. It also discusses the application prospect and existing problems of MALDI-MS in TCM， so as to provide technical support for the identification of active ingredients in TCM， drug utilization and metabolism.

Matrix-assisted laser desorption ionization mass spectrometry based quantitative analysis of cordycepin from Cordyceps militaris / 药物分析学报

Cordycepin,which has great immunomodulatory activities such as anticancer,antifungal,antivirus,antileukemia and lipid-lowering ones,is the secondary metabolite of Cordyceps militaris (C.militaris).Liquid submerged fermentation is the common cultivation process to produce cordycepin.To optimize the fermentation process and improve production,monitoring the cordycepin secretion in the fermen-tation is essential.The measurement based on chromatography-mass spectrometry methods is generally involved in the complex sample pretreatments and time-consuming separation,so more rapid and convenient methods are required.Matrix-assisted laser desorption ionization mass spectrometry(MALDI-MS) is more attractive for faster and direct detection.Therefore,MALDI-MS detection combined with isotope-labeled internal standard was applied to the measurement of cordycepin content in the fermentation broth and mycelium.This method made accurate quantification of cordycepin in the range of 5-400 μg/mL with a relative standard deviation of 5.6％.The recovery rates of fermentation samples after the 1,13,and 25 days were 90.15％,94.27％,and 95.06％,respectively.The contents of cordycepin in the mycelium and fermentation broth were 136 mg/g and 148.39 mg/mL on the 20th culture day,respectively.The cordycepin secretion curve of the liquid fermentation of C.militaris was real-time traced over 25 days.

Fast saccharide mapping method for quality consistency evaluation of commercial xylooligosaccharides collected in China / 药物分析学报

Due to the extensive use of xylooligosaccharides (XOS) as functional food ingredients,many inferior goods and even adulterants are generally found in the market,which may pose a health hazard to certain populations.Chromatography method such as high-performance liquid chromatography (HPLC) and high-performance thin-layer chromatography (HPTLC) is traditionally applied for the quality analysis of XOS.However,it is time consuming due to the prolonged separation and pre-or post-derivatization procedure.In this study,a fast saccharide mapping method based on matrix-assisted laser desorption/time-of-flight mass spectrometry (MALDI-TOF-MS) was developed for the quality consistency analysis of 22 batches of XOS collected from different manufacturers in China.The time needed for saccharides analysis using MALDI-MS was less than 30 min for one plate,at least 6 times faster than that by the traditional HPTLC chromatography method.In addition,MALDI-MS possessed higher resolution for XOS with DP4-DP7 based on the difference of m/z,which is hardly separated using HPTLC.The results showed that XOS were present only in samples XY01-XY11,samples XY12-XY14 only consisted of hex oligo-saccharides,and samples XY15-XY22 were free of oligosaccharides.These indicate that the quality consistency of XOS products in the China market was poor,which should be carefully investigated.

Candida auris infection in the central catheter of a patient without sepsis symptoms / Infección por Candida auris en el catéter central de un paciente sin síntomas de sepsis

Abstract Background: Candida auris is an emerging yeast frequently reported as resistant to multiple antifungal drugs commonly used to treat Candida infections. This specie can colonize the patient's skin and has great ability for producing outbreaks in hospitals. C. auris is phylogenetically related to other Candida species, can be misidentified using conventional biochemical or commercial methods and requires specific technology for its identification. Case report: We report the first isolate of C. auris in Cali, Colombia, from a central venous catheter in a 37-year-old patient with rheumatoid arthritis and endocarditis who did not have symptoms of sepsis. The yeast was initially misidentified as C. haemulonii using the Phoenix system and subsequently identified as C. auris by matrix-assisted laser desorption/ionization-time-of-flight mass spectrometry (MALDI-TOF MS). The broth microdilution method was used to determine the minimum inhibitory concentration; the isolate was susceptible to fluconazole, itraconazole, voriconazole and amphotericin B. Conclusions: This report contributes to knowledge of the epidemiology of C. auris infections in individuals with underlying disease and describes an isolate with a behavior different from what is usually reported.

Resumen Antecedentes: Candida auris es una levadura emergente, informada con frecuencia como resistente a diversos antifúngicos usados comúnmente para tratar infecciones por Candida. Esta especie puede colonizar la piel y tiene gran capacidad de producir brotes en ambientes hospitalarios. Está filogenéticamente relacionada con otras especies de Candida, es mal identificada por los métodos bioquímicos o comerciales, y requiere tecnología específica para su identificación. Reporte de caso: Se informa el primer aislamiento de C. auris en Cali, Colombia en un paciente de 37 años con artritis reumatoide y endocarditis, sin síntomas de sepsis, a partir de la punta de catéter venoso central. La levadura inicialmente se identificó como C. haemulonii por el sistema Phoenix® y posteriormente como C. auris por espectrometría de masas desorción/ionización láser asistida por una matriz con detección de masas por tiempo de vuelo (MALDI-TOF MS). Se determinó la concentración inhibitoria mínima por el método de microdilución en caldo que mostró un aislamiento sensible a fluconazol, itraconazol, voriconazol y anfotericina B. Conclusión: Este informe contribuye al conocimiento de la epidemiología de las infecciones por C. auris en individuos con enfermedad subyacente y describe un aislamiento con un comportamiento diferente a lo indicado en otros estudios.

Analytical capabilities of matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) and its potential applications in medicinal plants / 药学学报

Matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI), as a label-free imaging technique with high coverage and sensitivity is widely used for visualizing the spatial distribution of proteins, peptides and small metabolites in tissues. With the development of MALDI technique, MALDI-MSI is also employed to monitor the spatial distribution of phytochemical constituents of medicinal plants. In this review, we first briefly introduce MALDI-MSI technique, and we focus on its application in the spatial distribution and accumulation of secondary metabolites in medicinal plants. The ultimate advantage of using MALDI-MSI for spatial distribution analysis at the molecular level, offers crucial evidence of synthesis, transfer and accumulation of bioactive molecules in medicinal plants.

Rhein:a novel matrix for profiling and imaging of endogenous metabolites by matrix-assisted laser desorption/ionization-mass spectrometry / 中国药科大学学报

@#MALDI-MS is an emerging technique that enables rapid profiling of endogenous substances in a high throughput. Nevertheless, its application to metabolite analysis is often hindered by the presence of massive matrix background peaks using conventional matrices. Herein rhein was introduced as a novel matrix for MALDI-MS analysis of cation metabolites. Several strong and weak basic metabolites were measured using rhein. Compared to the previously reported ionless matrix NSA and conventional matrix CHCA, clean MALDI-MS spectra were obtained devoid of matrix signals. Subsequently, we applied this matrix to profile a biologically complex sample, mice intestinal contents. In addition, we have validated the applicability of rhein to MALDI-MS imaging, which allows for simultaneous mapping of hundreds of metabolites from single mouse ileum section. In summary, the discovery of rhein as a novel MALDI matrix enables efficient and reliable profiling and imaging of endogenous metabolites from biological samples with no matrix interferences, which will greatly promote the utility of MALDI-MS-based platform for metabolome studies.

Lipid structural information from a single equine embryo by MALDI-MS / Informação estrutural de lipídios a partir de um único embrião equino pela técnica de MALDI-MS

It was reported the potential of MALDI-MS for the characterization of lipid species present in a single equine embryo, and studied some lipid structures detected by collision induced dissociation (CID) experiments. In the positive ion mode spectrum, it were observed mostly protonated and sodiated species of sphingomyelins (SM), phosphatidylcholines (PC) and triacylglycerols (TAG). In the negative ion mode, it were observed phosphatidylethanolamines (PE) and phosphatidylinositols (PI). MS/MS spectrum of most intense lipid ions was performed to show MALDI-MS/MS structural information potential. MS/MS spectrum in the positive mode of m/z 760.6 (attributed as PC34:1) depicted characteristic PC fragments of m/z 184.1 (choline polar head), and the neutral loss (NL) of 183 (phosphorylcholine). For the ion of m/z 766.6 (attributed as PE 38:5), we observed the NL of 140, characteristic of PE. For the ion of m/z 808.7 (attributed as PC 38.5), besides the fragment at m/z 184.1 at the NL of 183, it was possible to observe the loss of trimethylamine (ion of m/z 749.6), and the cyclophosphane (ion of m/z 147.0). Finally, for the negative ion mode, we isolated and fragmented the ion at m/z 863.6, which was attributed as PI 36:1 due to the presence of m/z 153 (glycerol phosphate – H2 O-H), 223 (phospho inositol – 2H2 O-H), 241 (phospho inositol – H2 O-H), 281 (oleic acid), and 581.3 (lysophosphoinositol – H2 O-H). It was concluded that MALDI-MS allowed the detection of a broad range of PC, SM, PE, PI and TAG lipid species, as well as a fast and confident characterization of lipid structures from a single equine embryo.

É relatado o potencial da técnica de MALDI-MS para caracterizar espécies de lipídios presentes em um único embrião equino e estudadas algumas estruturas lipídicas detectadas por dissociação induzida por colisão (CID). No espectro de modo íon positivo, foram observadas espécies, principalmente, protonadas e sodiadas de esfingomielinas (SM), fosfatidileolinas (PC) e triacilgliceróis (TAG). No modo negativo, foram observadas fosfatidiletanolaminas (PE) e fosfatidilinositos (PI). Espectros de íons de lípidos com maior intensidade foram utilizados para demonstrar o potencial da informação estrutural por MALDI-MS/MS. O espectro no modo positivo de m/z (massa sobre carga) 760,6 (atribuída como PC34:1) apresentou características de fragmentos PC de m/z 184,1 (denominada cabeça polar de colina), além de perda neutral (NL) de m/z 183 (fosforilcolina). Para o íon de m/z 766,6 (atribuída como PE38:5), observou-se a NL de 140, característica do PE. Para o íon de m/z 808,7 (38,5 atribuído como PC), além do fragmento m/z 184,1 na NL de 183, foi possível observar a perda de trimetilamina (íon de m/z 749,6) e o ciclofosfano (íon de m/z 147,0). Finalmente, para o modo de íon negativo, foram isolados e fragmentados o íon de m/z 863,6 que foi atribuído como PI36:1, devido à presença de m/z 153 (fosfato de glicerol – H2 O-H ), 223 (inositol fosfo - 2H2 O-H) , 241 (fosfoinositol – H2 O-H), 281 (ácido oleico) e 581,3 (lisofosfoinositol – H2 O+H). Foi concluído que a MALDI - MS permite a detecção de uma ampla gama de espécies de PC, SM, PE, PI e TAG lipídicas, bem como a caracterização rápida e confiante de estruturas lipídicas a partir de um único embrião equino.

Plasma proteomic analysis of patients with squamous cell carcinoma of the uterine cervix / 부인종양

OBJECTIVE: To compare plasma protein expression between patients with squamous cell carcinoma (SCC) of the cervix and normal controls. METHODS: Plasma samples from patients with benign gynecological disease (normal cervix, n=6) and cervical cancer (SCC, n=6) were subjected to plasma proteomic analysis using two dimensional gel electrophoresis (2-DE) and matrix-assisted laser desorption/ionization mass spectroscopy (MALDI-MS). Western blotting and immunoturbidimetric assay were performed to validate the results of 2-DE. RESULTS: Eight proteins showed differential expression between controls and SCC patients; six (ceruloplasmin, complement C3, afamin precursor, alpha-1-B-glycoprotein, transferrin, alpha-fibrinogen precursor) were up-regulated, while two (chain A, crystal structure of antithrombin and apolipoprotein A-IV precursor) were down-regulated in the plasma of SCC patients. Western blotting analysis revealed significant elevation of ceruloplasmin, complement C3, afamin, and alpha-1-B-glycoprotein in the plasma of SCC patients in comparison to controls. Immunoturbidimetric assay of a larger group confirmed the results of 2-DE and Western blotting, and showed that ceruloplasmin and complement C3 were significantly elevated in the plasma of SCC patients in comparison with controls and patients with carcinoma in situ (CIS) of the uterine cervix. CONCLUSION: Plasma protein expression determined using 2-DE and MALDI-MS will give a chance to identify tumor-specific biomarkers for SCC of the cervix.

Analysis of the Cell Lysate Proteome of a Korean Mycobacterium tuberculosis Isolate K01 with H37Rv and H37Ra Strains

Despite recent economic prosperity, Korea still has high prevalence of tuberculosis. Molecular biologic characterization of Korean Mycobacterium tuberculosis strains might provide a deeper understanding of the forces contributing to the spread of tuberculosis in Korea. Therefore, we analyzed the cell lysate proteome of a representative Korean Mycobacterium tuberculosis isolate (K01) in comparison with laboratory reference strains H37Rv and H37Ra. Seven spots were strongly expressed only in K01 strain compared with M. tuberculosis H37Rv and H37Ra. Through continuous MALDI-MS analysis, these spots were identified as hypothetical protein Rv3849, secreted immunogenic protein Mpt64, Acetyl/propionyl-CoA Carbpxylase (AccD1), alkyl hydroperoxide reductase C (AhpC), N-acetylmuramyl-L-alanine amidase, a putative UDP glucose epimerase, and a transposase. A deeper study of these proteins may provide a clue in the development of effective new anti-tuberculosis vaccines against Korean M. tuberculosis isolates.