Search

The article focuses on the application of MALDI-TOF Mass Spectrometry in molecular biology diagnostics, particularly for the sensitive detection of single nucleotide variations (SNPs) in a multiplex approach. Topics include the principle of MALDI-TOF, its versatility in detecting genomic DNA or cDNA mutations, its role in thrombophilia diagnostics, testing before fluorouracil-based therapy, and its use in SARS-CoV-2 variant typing.

Introduction: Paenibacillus stellifer is widely distributed in nature, but its pathogenicity has not been reported since it was first identified in 2003. Objectives: This work aimed to identify Gram-variable rods isolated from cases of health care-associated infections in a hospital in a mountainous region of Rio de Janeiro, Brazil, between September/2015 and August/2016, and analyze their sensitivity to antibiotics commonly used in clinical practice. Methods: Initially, microorganisms were identified with conventional methods and confirmed by the Matrix-Assisted Laser Desorption/Ionization - Time of Flight (MALDI-TOF) technique. The sensitivity to antimicrobials test was performed according to recommendations from the Clinical and Laboratory Standards Institute. Results: We analyzed 105 samples: 59 surgical wound secretions and 46 blood cultures. Gram-variable rods were identified in two surgical wound secretion samples (3.39%) and two blood cultures (4.35%). Paenibacillus stellifer was the microorganism isolated from the four samples, showing sensitivity to all tested drugs. Conclusion: P. stellifer is a microorganism that is not in the make-up of human microbiota and has an environmental origin. According to current knowledge, this is the first identification of P. stellifer as the etiological agent of surgical wound infections in the world, and bacteremia in Brazil. Lastly, we highlight that microorganisms normally found in the environment are able to cause infections in a hospital. [ABSTRACT FROM AUTHOR]

Recently, the proteomic analysis has become an ideal tool to study the structure and function of platelets. We proposed a nano-liquid chromatography (nano-LC) technique coupled off-line with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF-MS) for rat platelet proteome analysis. In this study, we attempted to analyze the rat platelet proteome in two different subcellular fractions: cytosol and membrane. Platelet-rich plasma was collected from healthy rats. The platelet samples were extracted with Subcellular Proteome Extraction Kit to collect subcellular compartments. For further investigations, platelet lysate, cytosol and membrane fractions were used. Enzymatic digestion of proteins was performed using Filter Aided Sample Preparation method with trypsin as a proteolytic enzyme. Tryptic peptides were analyzed using nano-LC-MALDI-TOF/TOF-MS. Platelet proteins identification was performed using the Mascot engine. We identified 238 proteins in the platelet lysate, 210 in the cytosol, and 148 in the membrane fraction. Among them, 45 were unique for platelet lysate, 55 for cytosol, and 34 for the membrane fraction. The gene ontology analysis showed that there were differences in the proteome of cytosol and membrane fractions related to the molecular functions, i.e. coagulative activity. Our results may suggest that the membrane or cytosol location of the proteins with coagulative activity may be responsible for the acute or delayed platelet response to an agonist. The nano-LC-MALDI-TOF/TOF-MS method can be used for identifying proteins of subcellular fraction in rat platelets. [ABSTRACT FROM AUTHOR]

In this work, SiO 2 -IDA microspheres were obtained via a previous method by grafting iminodiacetic acid (IDA) groups onto silica microspheres, then the SiO 2 -IDA microspheres were packed in capillary columns to prepare SiO 2 -IDA capillary trap columns for the first time. Based on the trap columns, a novel on-column enrichment method toward glycopeptides was developed as a sample pretreatment step prior to matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF MS) or nano-LC-MS/MS analysis. The preparation procedures of the trap columns are well reproducible. Thanks to outstanding hydrophilicity brought by the grafted zwitterionic IDA groups, the trap columns showed excellent on-column enrichment performance toward glycopeptides by hydrophilic interaction liquid chromatography (HILIC). Experimental results showed that the on-column enrichment method could reach a low detection limit (50 fmol HRP digest) and a high selectivity (molar ratio of HRP digest and BSA digest 1:100). Finally, it was applied to on-column enrichment of glycopeptides from digests of human serum followed by nano-LC-MS/MS analysis. In total, 207 glycopeptides assigned to 78 glycoproteins were identified from 2 μL human serum. [ABSTRACT FROM AUTHOR]

The Bruker MALDI Biotyper® method utilizes matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) MS for the rapid and accurate identification and confirmation of Gram-negative bacteria from select media types. The alternative method was evaluated using nonselective and selective agars to identify Cronobacter spp., Salmonella spp., and select Gram-negative bacteria. Results obtained by the Bruker MALDI Biotyper were compared to the traditional biochemical methods as prescribed in the appropriate reference methods. Two collaborative studies were organized, one in the United States focusing on Cronobacter spp. and other Gram-negative bacteria, and one in Europe focusing on Salmonella spp. and other Gramnegative bacteria. Fourteen collaborators from seven laboratories located within the United States participated in the first collaborative study for Cronobacter spp. Fifteen collaborators from 15 service laboratories located within Europe participated in the second collaborative study for Salmonella spp. For each target organism (either Salmonella spp. or Cronobacter spp.), a total of 24 blind-coded isolates were evaluated. In each set of 24 organisms, there were 16 inclusivity organisms (Cronobacter spp. or Salmonella spp.) and 8 exclusivity organisms (closely related non-Cronobacter spp. and non-Salmonella spp. Gram-negative organisms). After testing was completed, the total percentage of correct identifications from each agar type for each strain was determined at a percentage of 100.0% to the genus level for the Cronobacter study and a percentage of 100.0% to the genus level for the Salmonella study. For both non-Cronobacter and non-Salmonella organisms, a percentage of 100.0% was correctly identified. The results indicated that the alternative method produced equivalent results when compared to the confirmatory procedures specified by each reference method. [ABSTRACT FROM AUTHOR]

Infectious bovine keratoconjunctivitis (IBK) is an economically significant disease caused by Moraxella bovis. Moraxella bovoculi, although not reported to cause IBK, has been isolated from the eyes of cattle diagnosed with IBK. Identification of M. bovis and M. bovoculi can be performed using biochemical or DNA-based approaches, both of which may be time consuming and inconsistent between laboratories. We conducted a comparative evaluation of M. bovoculi and M. bovis identification using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) with a database provided by Bruker Daltonics (termed the BDAL database), the BDAL database supplemented with spectra generated in our study (termed the UNLVDC database), and with PCR–restriction-fragment length polymorphism (PCR-RFLP) typing. M. bovoculi (n = 250) and M. bovis (n = 18) isolates from cattle with or without IBK were used. MALDI-TOF MS using the UNLVDC database correctly identified 250 of 250 (100%) of M. bovoculi and 17 of 18 (94%) of M. bovis isolates. With the BDAL database, MALDI-TOF MS correctly identified 249 of 250 (99%) of M. bovoculi and 7 of 18 (39%) of M. bovis isolates. In comparison, the PCR-RFLP test correctly identified 210 of 250 (84%) of M. bovoculi and 12 of 18 (66%) of M. bovis isolates. Thus, MALDI-TOF MS with the UNLVDC database was the most effective identification methodology for M. bovis and M. bovoculi isolates from cattle. [ABSTRACT FROM AUTHOR]

Since its introduction in the 1980s, matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS) has gained a prominent role in the analysis of high molecular weight biomolecules such as proteins, peptides, oligonucleotides, and polysaccharides. Its application to low molecular weight compounds has remained for long time challenging due to the spectral interferences produced by conventional organic matrices in the low m/z window. To overcome this problem, specific sample preparation such as analyte/matrix derivatization, addition of dopants, or sophisticated deposition technique especially useful for imaging experiments, have been proposed. Alternative approaches based on second generation (rationally designed) organic matrices, ionic liquids, and inorganic matrices, including metallic nanoparticles, have been the object of intense and continuous research efforts. Definite evidences are now provided that MALDI MS represents a powerful and invaluable analytical tool also for small molecules, including their quantification, thus opening new, exciting applications in metabolomics and imaging mass spectrometry. This review is intended to offer a concise critical overview of the most recent achievements about MALDI matrices capable of specifically address the challenging issue of small molecules analysis.An ideal Book of matrices for MALDI MS of small molecules [ABSTRACT FROM AUTHOR]

LC-MS/MS is widely utilized today for quantification of vitamin D in biological fluids. Mass spectrometric assays for vitamin D require very careful method optimization for precise and interference-free, accurate analyses however. Here, we explore chemical derivatization and matrix-assisted laser desorption/ionization (MALDI) as a rapid alternative for quantitative measurement of 25-hydroxyvitamin D3 in human serum, and compare it to results from LC-MS/MS. The method implemented an automated imaging step of each MALDI spot, to locate areas of high intensity, avoid sweet spot phenomena, and thus improve precision. There was no statistically significant difference in vitamin D quantification between the MALDI-MS/MS and LC-MS/MS: mean ± standard deviation for MALDI-MS—29.4 ± 10.3 ng/mL—versus LC-MS/MS—30.3 ± 11.2 ng/mL (P = 0.128)—for the sum of the 25-hydroxyvitamin D epimers. The MALDI-based assay avoided time-consuming chromatographic separation steps and was thus much faster than the LC-MS/MS assay. It also consumed less sample, required no organic solvents, and was readily automated. In this proof-of-concept study, MALDI-MS readily demonstrated its potential for mass spectrometric quantification of vitamin D compounds in biological fluids.ᅟ [ABSTRACT FROM AUTHOR]

Matrix-assisted laser/desorption ionization (MALDI) mass spectrometry imaging (MSI) is widely used as a unique tool to record the distribution of a large range of biomolecules in tissues. 2,6-Dihydroxyacetophenone (DHA) matrix has been shown to provide efficient ionization of lipids, especially gangliosides. The major drawback for DHA as it applies to MS imaging is that it sublimes under vacuum (low pressure) at the extended time necessary to complete both high spatial and mass resolution MSI studies of whole organs. To overcome the problem of sublimation, we used an atmospheric pressure (AP)-MALDI source to obtain high spatial resolution images of lipids in the brain using a high mass resolution mass spectrometer. Additionally, the advantages of atmospheric pressure and DHA for imaging gangliosides are highlighted. The imaging of [M-H]− and [M-H2O-H]− mass peaks for GD1 gangliosides showed different distribution, most likely reflecting the different spatial distribution of GD1a and GD1b species in the brain.ᅟ [ABSTRACT FROM AUTHOR]

Currently, the capability of identification for Acinetobacter species using MALDI-TOF MS still remains unclear in clinical laboratories due to certain elusory phenomena. Thus, we conducted this research to evaluate this technique and reveal the causes of misidentification. Briefly, a total of 788 Acinetobacter strains were collected and confirmed at the species level by 16S rDNA and rpoB sequencing, and subsequently compared to the identification by MALDI-TOF MS using direct smear and bacterial extraction pretreatments. Cluster analysis was performed based on the mass spectra and 16S rDNA to reflect the diversity among different species. Eventually, 19 Acinetobacter species were confirmed, including 6 species unavailable in Biotyper 3.0 database. Another novel species was observed, temporarily named A. corallinus. The accuracy of identification for Acinetobacter species using MALDI-TOF MS was 97.08% (765/788), regardless of which pretreatment was applied. The misidentification only occurred on 3 A. parvus strains and 20 strains of species unavailable in the database. The proportions of strains with identification score ≥ 2.000 using direct smear and bacterial extraction pretreatments were 86.04% (678/788) and 95.43% (752/788), χ2 = 41.336, P < 0.001. The species similar in 16 rDNA were discriminative from the mass spectra, such as A. baumannii & A. junii, A. pittii & A. calcoaceticus, and A. nosocomialis & A. seifertii. Therefore, using MALDI-TOF MS to identify Acinetobacter strains isolated from clinical samples was deemed reliable. Misidentification occurred occasionally due to the insufficiency of the database rather than sample extraction failure. We suggest gene sequencing should be performed when the identification score is under 2.000 even when using bacterial extraction pretreatment.ᅟ [ABSTRACT FROM AUTHOR]

We demonstrate that a donut-shaped surface pattern consisting of a central hydrophobic region and a surrounding hydrophilic region simultaneously concentrates and desalts a solution of neuropeptides with a high salt content. Our approach greatly simplifies the sample preparation process for MALDI mass spectrometry. [ABSTRACT FROM AUTHOR]

Whole cell MALDI is regularly used for the identification of bacteria to species level in clinical Microbiology laboratories. However, there remains a need to rapidly characterize and differentiate isolates below the species level to support outbreak management. We describe the implementation of a modified preparative approach for MALDI-MS combined with a custom analytical computational pipeline as a rapid procedure for subtyping Shigatoxigenic E. coli (STEC) and accurately identifying strain-specifying biomarkers. The technique was able to differentiate E. coli O157:H7 from other STEC. Within O157 serotype O157:H7 isolates were readily distinguishable from Sorbitol Fermenting O157 isolates. Overall, nine homogeneous groups of isolates were distinguished, each exhibiting distinct profiles of defining mass spectra features. This offers a robust analytical tool useable in reference/diagnostic public health scenarios. [ABSTRACT FROM AUTHOR]

Iron-oxide nanoparticles (NPs) generated by environmental events are likely to represent health problems. α-Fe 2 O 3 NPs were synthesized, characterized and tested in a model for toxicity utilizing human whole blood without added anticoagulant. MALDI-TOF of the corona was performed and activation markers for plasma cascade systems (complement, contact and coagulation systems), platelet consumption and release of growth factors, MPO, and chemokine/cytokines from blood cells were analyzed. The coronas formed on the pristine α-Fe 2 O 3 NPs contained contact system proteins and they induced massive activation of the contact (kinin/kallikrein) system, as well as thrombin generation, platelet activation, and release of two pro-angiogeneic growth factors: platelet-derived growth factor and vascular endothelial growth factor, whereas complement activation was unaffected. The α-Fe 2 O 3 NPs exhibited a noticeable toxicity, with kinin/kallikrein activation, which may be associated with hypotension and long-term angiogenesis in vivo, with implications for cancer, arteriosclerosis and pulmonary disease. [ABSTRACT FROM AUTHOR]

Ancient Egyptian painted artworks are usually understudied from an analytical point of view, due to their extremely fragile nature. Attention typically focuses on pigments since identification is possible with non-invasive techniques, while limited information is available in the literature regarding the organic binding media. Here successful determination of the binder of a Roman Egyptian painted mummy shroud (2nd–3rd century A.D.) achieved through the application of enzymatic digestion followed by matrix-assisted laser desorption ionization mass spectrometry (MALDI MS) is reported. The high specificity and sensitivity of this analytical strategy not only allowed the identification of the binding medium as a mixture of two different plant gums but also allowed the discrimination of the different species sources, even though the organic material was present in very small amounts and subject to degradation. The results of this study represent the first analytical identification of the earliest use of locust bean gum as a paint binder material as well as the use of gum arabic from an Acacia species different from the well-known Acacia senegal . The precise identification of the organic binder is a great step forward in the understanding of the painting materials and techniques used in Roman Egypt, of which little is known. The result of this research opens new avenues of art historical and conservation investigation into the specific plant sources and types selected by artists and it has implications for future conservation treatment options. [ABSTRACT FROM AUTHOR]

Copyright of Revista Española de Quimioterapia is the property of Sociedad Espanola de Quimioterapia and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

In this paper, we evaluated matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) as a cultivation-independent, routinely applicable approach to identify microbial fractions in bioaerosol emission samples. We developed a streamlined protocol in line with the German state-of-the-art impingement sampling guideline. Following isokinetic sampling, a fast and reliable pre-treatment methodology involving a series of cascade filtration steps was implemented, which produced fractions for spectrometric measurement devoid of interfering substances. We sampled the exhaust air from eight pig fattening farms around western Germany, which yielded two sets of samples for both method development and validation. For method development, in total 65 bacterial isolates were produced directly from the exhaust air samples, taxonomically classified by 16S rRNA-Gene sequencing, and subjected to MALDI-TOF analysis. In this way, we could assign fingerprint biomarkers to classified bacterial genera or even species to build up a preliminary reference database. For verification of the novel methodology and application of the reference database, we subjected the second set of exhaust air samples to the developed protocol. Here, 18 out of 21 bacterial species deposited in the database were successfully retrieved, including organisms classified in risk group 2, which might be used to evaluate the pathogenic potential of sampled exhaust air. Overall, this study pursues an entirely new approach to rapidly analyze airborne microbial fractions. [ABSTRACT FROM AUTHOR]

A new strategy to obtain IRMPD spectra of metal cationized ions by combining MALDI and IRMPD methods is presented here, in which graphene is selected to be the matrix to generate the metal cationized ions. [Arg + Rb] + were studied here as the sample ions. Based on the experiential IRMPD spectrum and theoretical calculations, it is suggested that the ions of [Arg + Rb] + generated here had more internal energies than those generated by ESI method reported previously by Williams et al. (J. Phys. Chem. A 2007, 111, 11759–11770). The method is also combined with the H/D exchange to identify the chromophores for the observed IR peak. The method provides a new way to obtain structural information of metal cationized ions, and also makes the identification of isomeric compounds feasible. [ABSTRACT FROM AUTHOR]

The analytical performances of the RAPIDEC CARBA NP® (bioMérieux) and the β-CARBA® (Bio-Rad) tests were evaluated for the rapid detection of any known type of carbapenemases in Enterobacteriaceae . An international collection of 149 enterobacterial isolates comprising 111 Carbapenemase-Producing Enterobacteriaceae (CPE) and 38 non-carbapenemase producers was used. CPE included 32 carbapenemase producers of class A (18 KPC-2, 1 FRI-1, 5 SME and 8 IMI), 33 of class B (13 NDM, 11 VIM and 9 IMP) and 46 of class D (15 OXA-48, 14 OXA-181, 10 OXA-204, 3 OXA-232 and 4 OXA-162). The RAPIDEC CARBA NP® and the β-CARBA® tests were performed in strict accordance to the manufacturer's instructions and results were read within 2 h and 30 min, respectively. RAPIDEC CARBA NP® detected 104/111 CPE isolates compared to 72/111 for the β-CARBA® test. The overall sensitivity and specificity were 93.7% and 100%, respectively, for the RAPIDEC CARBA NP® test, and 64.9% and 90%, respectively, for the β-CARBA® test. The β-CARBA® test failed to detect all the non-KPC class A carbapenemases (14/14) and most (24/31) of the OXA-48-like producers (OXA-162, OXA-181, OXA-204 and OXA-232), and detected 1/1 OXA-163 and 1/1 OXA-405 as carbapenemase producers whereas these enzymes are rather defined as non carbapenemases. RAPIDEC CARBA NP® test exhibited better performances than those of the β-CARBA® test and confirmed to be a reliable tool for the detection of CPE, especially for OXA-48-like producers. [ABSTRACT FROM AUTHOR]

The goat milk is considered as nutraceutical due to its several beneficial compounds that increase its value. The purpose of this study was to investigate the variation of the goat milk's lactic profile during winter and spring (2015). Several tests were realised. First, the isolation of the lactic acid bacteria contained in goat milk was realised by three specific media: the MRS, the M17, and the MSE. Second, a screening by the study of the proteolysis at two different concentration of casein (2%, 5%) and the study of aroma produc- tion were made to highlight performing isolates. These performing isolates were identified by using a precision spectrometry method which is called the Matrix Assisted Laser Desorption Ionisation Time-of-flight (MALDI-TOF). The identification with mass spectrometry «MALDI-TOF» technique revealed that those seasons had a more quantitative effect than a qualitative one. As a result, an average of 48% of lactic acid bacteria was identified for both seasons with a decrease of 6% observed in winter. In addition, it was revealed that Enterococcus was the most dominant genus in the Algerian goat milk. Therefore, lactic strains of this milk were proved to be technologically competent in terms of proteolysis and aroma production. [ABSTRACT FROM AUTHOR]

The Morris water maze (MWM) spatial learning task has been demonstrated to involve a cognitive switch of action control to serve the transition from an early towards a late learning phase. However, the molecular mechanisms governing this switch are largely unknown. We employed MALDI MS imaging (MSI) to screen for changes in expression of small proteins in brain structures implicated in the different learning phases. We compared mice trained for 3 days and 30 days in the MWM, reflecting an early and a late learning phase in relation to the acquisition of a spatial learning task. An ion with m/z of 6724, identified as PEP-19/pcp4 by top-down tandem MS, was detected at higher intensity in the dorsal striatum of the late learning phase group compared with the early learning phase group. In addition, mass spectrometric analysis of synaptosomes confirmed the presence of PEP-19/pcp4 at the synapse. PEP-19/pcp4 has previously been identified as a critical determinant of synaptic plasticity in locomotor learning. Our findings extend PEP-19/pcp4 function to spatial learning in the forebrain and put MSI forward as a valid and unbiased research strategy for the discovery and identification of the molecular machinery involved in learning, memory and synaptic plasticity. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann. [ABSTRACT FROM AUTHOR]

Matrix-assisted laser desorption/ionization (MALDI)-mass spectrometry imaging (MSI) permits label-free in situ analysis of chemical compounds directly from the surface of two-dimensional biological tissue slices. It links qualitative molecular information of compounds to their spatial coordinates and distribution within the investigated tissue. MALDI-MSI can also provide the quantitative amounts of target compounds in the tissue, if proper calibration techniques are performed. Obviously, as the target molecules are embedded within the biological tissue environment and analysis must be performed at their precise locations, there is no possibility for extensive sample clean-up routines or chromatographic separations as usually performed with homogenized biological materials; ion suppression phenomena therefore become a critical side effect of MALDI-MSI. Absolute quantification by MALDI-MSI should provide an accurate value of the concentration/amount of the compound of interest in relatively small, well-defined region of interest of the examined tissue, ideally in a single pixel. This goal is extremely challenging and will not only depend on the technical possibilities and limitations of the MSI instrument hardware, but equally on the chosen calibration/standardization strategy. These strategies are the main focus of this article and are discussed and contrasted in detail in this tutorial review. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann. [ABSTRACT FROM AUTHOR]

In this work, we combined the use of two MALDI matrices (quercetin and 9-aminoacridine), a recently developed new matrix coating technique – matrix coating assisted by an electric field (MCAEF), and matrix-assisted laser desorption/ionization – Fourier transform ion cyclotron resonance mass spectrometry (MALDI-FTICRMS) to detect and image endogenous compounds in the cancerous and non-cancerous regions of three human prostate cancer (stage II) tissue specimens. After three rounds of imaging data acquisitions (i.e., quercetin for positive and negative ion detection and 9-aminoacridine for negative ion detection), and metabolite identification, a total of 1091 metabolites including 1032 lipids and 59 other metabolites were routinely detected and successfully localized. Of these compounds, 250 and 217 were only detected in either the cancerous or the non-cancerous regions respectively, although we cannot rule out the presence of these metabolites at concentrations below the detection limit. In addition, 152 of the other 624 metabolites showed differential distributions ( p < 0.05, t -test) between the two regions of the tissues. Further studies on a larger number of clinical specimens will need to be carried out to confirm this large number of apparently cancer-related metabolites. The successful determination of the spatial locations and abundances of these endogenous biomolecules indicated significant metabolism abnormalities – e.g., increased energy charge and under-expression of neutral acyl glycerides, in the prostate cancer samples. To our knowledge, this work has resulted in MALDI-MS imaging of the largest group of metabolites in prostate cancer thus far and demonstrated the importance of using complementary matrices for comprehensive metabolomic imaging by MALDI-MS. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann. [ABSTRACT FROM AUTHOR]

An integrated diagnosis using molecular features is recommended in the 2016 World Health Organization (WHO) classification. Our aim was to explore non-targeted molecular classification using MALDI mass spectrometry imaging (MALDI MSI) associated to microproteomics in order to classify anaplastic glioma by integration of clinical data. We used fresh-frozen tissue sections to perform MALDI MSI of proteins based on their digestion peptides after in-situ trypsin digestion of the tissue sections and matrix deposition by micro-spraying. The generated 70 μm spatial resolution image datasets were further processed by individual or global segmentation in order to cluster the tissues according to their molecular protein signature. The clustering gives 3 main distinct groups. Within the tissues the ROIs (regions of interest) defined by these groups were used for microproteomics by micro-extraction of the tryptic peptides after on-tissue enzymatic digestion. More than 2500 proteins including 22 alternative proteins (AltProt) are identified by the Shotgun microproteomics. Statistical analysis on the basis of the label free quantification of the proteins shows a similar classification to the MALDI MSI segmentation into 3 groups. Functional analysis performed on each group reveals sub-networks related to neoplasia for group 1, glioma with inflammation for group 2 and neurogenesis for group 3. This demonstrates the interest on these new non-targeted large molecular data combining both MALDI MSI and microproteomics data, for tumor classification. This analysis provides new insights into grade III glioma organization. This specific information could allow a more accurate classification of the biopsies according to the prognosis and the identification of potential new targeted therapeutic options. This article is part of a Special Issue entitled: MALDI Imaging, edited by Dr. Corinna Henkel and Prof. Peter Hoffmann. [ABSTRACT FROM AUTHOR]

The disulfide bond is one of the most common post-translational modifications in proteins, of which determination is essential to the comprehensive understanding of protein structures. Disulfide bond analysis has gone through great improvement due to the development of matrix-assisted laser desorption/ionization mass spectrometry (MALDI MS), especially in terms of speed and sensitivity. In general, the characterization of disulfide-containing peptides is achieved by the reduction of disulfide bonds followed by alkylation. The identification of disulfide/cysteine-containing peptides in digests of proteins is essential to structure elucidation of a protein. In order to present a deep understanding of some phenomena occurring in MALDI MS of disulfide/cysteine-containing proteins, the current work systematically investigated effects of co-crystal size and matrix on MALDI-In Source Decay (ISD) fragmentation of disulfide-containing proteins. Imaging experiments were performed to evaluate the influence of laser shot location on the fragmentation of human insulin and insulin glargine, which were selected as model compounds. The spectrum that was recorded on very finely distributed crystals of FA spot does not exhibit fragments. While a characteristic ‘triplet’ ions with a mass separation of 32 u generated by both symmetric and nonsymmetric cleavages of the disulfide bonds was observed on large crystals. Probably for thermodynamic reasons, the tryptic peptide was subjected to the cleavage of even number of chemical bonds gave rise to radical recombination without reductive ISD. Among several matrices tested including ferulic acid (FA), α-cyano-4-hydroxycinnamic acid (CHCA), sinapinic acid (SA), 2,5-dihydroxybenzoic acid (DHB), 3-aminoquinolin (AQ), and 2,4,6-trihydroxy acetophenone (THAP), FA was shown to be a versatile matrix allowing one to induce or prevent ISD according to the location of laser shots. CHCA and SA were found to promote ISD of disulfide-containing proteins, in a location dependent manner. However, unlike in CHCA (or SA), ISD was not systematically observed on all crystals for FA, suggesting differences between the crystallization processes of CHCA (or SA) and FA. Minor fragments were observed when using DHB and AQ as matrices. As for THAP, no fragmentation was observed probably because its three OH-groups in meta-position relative to each other resulted in the nonoccurrence of redox reaction. The studies provide insights into the experimental conditions required for determination of disulfide-containing protein by MALDI MS and are helpful for mass spectrum interpretation, opening the way to more rational studies of disulfide/cysteine-containing proteins by MALDI mass spectrometry. [ABSTRACT FROM AUTHOR]

The present work demonstrates that yeasts belonging to the Schizosaccharomyces genus release a high quantity of polysaccharides of cell wall origin starting from the onset of the alcoholic fermentation. By the end of the alcoholic fermentation, all of the Schizosaccharomyces yeast strains released a quantity of polysaccharides approximately 3–7 times higher than that released by a commercial Saccharomyces cerevisiae yeast strain under the same fermentative conditions of synthetic juice. A higher content of polysaccharide was found in media fermented by Schizosaccharomyces japonicus with respect to that of Schizosaccharomyces pombe . Some of the strains evaluated were also able to produce high levels of pyruvic acid, which has been shown to be an important compound for color stability of wine. The presence of strains with different malic acid consumption patterns along with high polysaccharide release would enable production of naturally modified wines with enhanced mouth feel and reduced acidity. The chemical analysis of the released polysaccharides demonstrated divergence between the two yeast species S. pombe and S. japonicus . A different mannose/galactose ratio and a different percentage of proteins was observed on the polysaccharides released by S. pombe as compared to S. japonicus . Analysis of the proteins released in the media revealed the presence of a glycoprotein with a molecular size around 32–33 kDa only for the species S. japonicus . Mass spectrometry analysis of carbohydrate moieties showed similar proportions among the N -glycan chains released in the media by both yeast species but differences between the two species were also observed. These observations suggest a possible role of rapid MALDI-TOF screening of N -glycans compositional fingerprint as a taxonomic tool for this genus. Polysaccharides release in the media, in particular galactomannoproteins in significant amounts, could make these yeasts particularly interesting also for the industrial production of exogenous polysaccharide preparations. [ABSTRACT FROM AUTHOR]

Salinity stress causes adverse physiological and biochemical changes in the growth and productivity of a plant. Azolla, a symbiotic pteridophyte and potent candidate for biofertilizer due to its nitrogen fixation ability, shows reduced growth and nitrogen fixation during saline stress. To better understand regulatory components involved in salinity-induced physiological changes, in the present study, Azolla microphylla plants were exposed to NaCl (6.74 and 8.61 ds/m) and growth, photochemical reactions of photosynthesis, ion accumulation, and changes in cellular proteome were studied. Maximum dry weight was accumulated in control and untreated plant while a substantial decrease in dry weight was observed in the plants exposed to salinity. Exposure of the organism to different concentrations of salt in hydroponic conditions resulted in differential level of Na and K ion accumulation. Comparative analysis of salinity-induced proteome changes in A. microphylla revealed 58 salt responsive proteins which were differentially expressed during the salt exposure. Moreover, 42 % spots among differentially expressed proteins were involved in different signaling events. The identified proteins are involved in photosynthesis, energy metabolism, amino acid biosynthesis, protein synthesis, and defense. Downregulation of these key metabolic proteins appears to inhibit the growth of A. microphylla in response to salinity. Altogether, the study revealed that in Azolla, increased salinity primarily affected signaling and photosynthesis that in turn leads to reduced biomass. [ABSTRACT FROM AUTHOR]

To improve the lateral resolution in matrix-assisted laser desorption/ionization mass spectrometry imaging (MALDI-MSI) beyond the dimensions of the focal laser spot oversampling techniques are employed. However, few data are available on the effect of the laser spot size and its focal beam profile on the ion signals recorded in oversampling mode. To investigate these dependencies, we produced 2 times six spots with dimensions between ~30 and 200 μm. By optional use of a fundamental beam shaper, square flat-top and Gaussian beam profiles were compared. MALDI-MSI data were collected using a fixed pixel size of 20 μm and both pixel-by-pixel and continuous raster oversampling modes on a QSTAR mass spectrometer. Coronal mouse brain sections coated with 2,5-dihydroxybenzoic acid matrix were used as primary test systems. Sizably higher phospholipid ion signals were produced with laser spots exceeding a dimension of ~100 μm, although the same amount of material was essentially ablated from the 20 μm-wide oversampling pixel at all spot size settings. Only on white matter areas of the brain these effects were less apparent to absent. Scanning electron microscopy images showed that these findings can presumably be attributed to different matrix morphologies depending on tissue type. We propose that a transition in the material ejection mechanisms from a molecular desorption at large to ablation at smaller spot sizes and a concomitant reduction in ion yields may be responsible for the observed spot size effects. The combined results indicate a complex interplay between tissue type, matrix crystallization, and laser-derived desorption/ablation and finally analyte ionization. [ABSTRACT FROM AUTHOR]

Matrix-assisted laser desorption/ionization imaging of biofilms cultured on agar plates is challenging because of problems related tomatrix deposition onto agar.We describe a one-step, spray-based application of a 2,5-dihydroxybenzoic acid solution for direct matrix-assisted laser desorption/ionization imaging of hydrated Bacillus subtilis biofilms on agar. Using both an optimized airbrush and a home-built automatic sprayer, region-specific distributions of signaling metabolites and cannibalistic factors were visualized from B. subtilis cells cultivated on biofilm-promoting medium. The approach provides a homogeneous, relatively dry coating on hydrated samples, improving spot to spot signal repeatability compared with sieved matrix application, and is easily adapted for imaging a range of agar-based biofilms. [ABSTRACT FROM AUTHOR]

1. The aim of this study was to evaluate the occurrence ofCampylobacterspp. in domestic and free-living pigeons and to evaluate the antibiotic resistance profiles. 2. The material consisted of cloacal swabs obtained from 108 homing pigeons and fresh faeces from 72 wild birds from Lublin and its vicinity. The identification of strains isolated on differential/selective media forCampylobacterspp. was carried out by MALDI-TOF and PCR. The susceptibility to antibiotics was evaluated by minimum inhibitory concentration (MIC) in Mueller-Hinton broth. 3. A total of 35 strains ofCampylobacterspp. were isolated; 27 were identified asCampylobacter jejuniand 8 asCampylobacter coli. Over half of the isolates were resistant to erythromycin and streptomycin, 40% of strains were resistant to tetracycline and ampicillin and 37% isolates were resistant to amoxicillin. Resistance to two or more antibiotics was observed in all strains tested. 4. The results indicate that both domestic and free-living pigeons are reservoirs for bacteria of the genusCampylobacter, which are characterised by varied and growing resistance to commonly used antibiotics. [ABSTRACT FROM PUBLISHER]

Streptococci are common causes of infective endocarditis (IE) and matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has provided a practical tool for their species determination. We aimed to investigate if particular groups of non-beta heamolytic streptococci were associated with IE or to specific presentations thereof. The Swedish Registry of Infective Endocarditis was used to identify cases of IE caused by streptococci and a local database to identify cases of streptococcal bacteremia. The bacteria were grouped using MALDI-TOF MS and the clinical characteristics of IE caused by different groups were compared. We identified a group of 201 streptococcal IE isolates: 18 isolates belonged to the anginosus, 19 to the bovis, 140 to the mitis, 17 to the mutans, and seven to the salivarius groups. The mitis and mutans groups were significantly more common and the anginosus group less common among IE cases as compared to all cause bacteremia. Patients infected with the bovis group isolates were older, had more cardiac devices, and had more commonly prosthetic valve IE compared to IE caused by streptococci of the other groups. Twenty-one percent of patients needed surgery, and in-hospital mortality was 8% with no significant differences between the groups. Grouping of non-beta haemolytic streptococci using MALDI-TOF MS can provide a basis for decision-making in streptococcal bacteremia. IE caused by bovis group isolates have clinical characteristics distinguishing them from IE caused by other groups of Streptococcus. [ABSTRACT FROM AUTHOR]

Using cross-linking coupled to matrix-assisted laser desorption/ionization mass spectrometry and CLIP-Seq sequencing, we determined the peptide and oligonucleotide sequences at the interfaces between the capsid proteins and the genomic RNA of bacteriophage MS2. The results suggest that the same coat protein (CP)–RNA and maturation protein (MP)–RNA interfaces are used in every viral particle. The portions of the viral RNA in contact with CP subunits span the genome, consistent with a large number of discrete and similar contacts within each particle. Many of these sites match previous predictions of the locations of multiple, dispersed and degenerate RNA sites with cognate CP affinity termed packaging signals (PSs). Chemical RNA footprinting was used to compare the secondary structures of protein-free genomic fragments and the RNA in the virion. Some PSs are partially present in protein-free RNA but others would need to refold from their dominant solution conformations to form the contacts identified in the virion. The RNA-binding peptides within the MP map to two sections of the N-terminal half of the protein. Comparison of MP sequences from related phages suggests a similar arrangement of RNA-binding sites, although these N-terminal regions have only limited sequence conservation. In contrast, the sequences of the C-termini are highly conserved, consistent with them encompassing pilin-binding domains required for initial contact with host cells. These results provide independent and unambiguous support for the assembly of MS2 virions via a PS-mediated mechanism involving a series of induced-fit viral protein interactions with RNA. [ABSTRACT FROM AUTHOR]

Surveillance of Streptococcus pneumoniae serotypes is important for the successful implementation of vaccination strategies to prevent the spread of invasive pneumococcal diseases. The standard method of serotyping of pneumococcal isolates is the phenotypic Neufeld test, which is cost- and labor-intensive. Recently, matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) has been implemented as a rapid, simple and inexpensive method for identifying species. We evaluated the performance of MALDI-TOF MS for serotyping ten major serotypes of S. pneumoniae in Japan (serotypes 3, 6B, 15A, 15C, 19A, 19 F, 23A, 24 F, 35B and 38) using the Biotyper and ClinProTools. After optimizing the settings, we validated their serotyping performance for serotypes 3, 15A and 19A using a separate set of isolates that were not used in the creation of the classification algorithms. A total of 574 isolates of S. pneumoniae collected from Japanese nationwide surveillance studies were included. Of these, 407 isolates belonged to the ten major serotypes. Biotyper and ClinProTools correctly identified 77.9 % and 84.0 %, respectively, of the ten major serotype isolates. The validation analysis included a total of 113 isolates of the serotypes 3, 15A and 19A isolates. Biotyper and ClinProTools correctly identified 85.0 % and 69.9 % of the validation cohort isolates, respectively. MALDI-TOF MS has the potential to discriminate the ten major S. pneumoniae serotypes prevalent in Japan. [ABSTRACT FROM AUTHOR]

Background: Pasteurella multocida meningitis in an immunocompetent patient is rare and commonly occurs after animal bite. To our knowledge, only 48 cases have been reported in the literature since 1989. P. multocida meningitis is commonly linked to animal contagion. Here we report on a new case of P. multocida meningitis in an immunocompetent patient who is a dog owner without a dog bite. We used the matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry to investigate the clonal lineage between animal and human isolates. Case presentation: In our case, a 25-year-old immunocompetent French Caucasian woman with nothing notable in her medical history was admitted for meningitis caused by P. multocida. Clonal lineage of P. multocida strains from cerebrospinal fluid and blood culture and her dog's oral cavity has been recognized by MALDI-TOF mass spectrometry dendrograms and clustering of the 21 P. multocida isolates in our centres. She was treated by a combination of intravenous ceftriaxone (2 g/day) and oral levofloxacin (1 g/day). She was discharged on the 6th day of admission. The antimicrobial therapy was conducted for 15 days. The dog was treated by clavulanic-acid amoxicillin for 3 weeks by the veterinarian. The evolution of the patient at the 5th month after the end of the antimicrobial therapy was normal without any neurological after-effects. Conclusion: The meningitis caused by P. multocida could be considered a cause of human meningitis in dog lovers without an animal bite. MALDI-TOF mass spectrometry should be considered as it is an accurate tool to identify clonal lineage between animal and human isolates. [ABSTRACT FROM AUTHOR]

The article is a study on the use of an achromatic objective lens as a versatile imaging interface for high-lateral-resolution atmospheric pressure (AP) matrix assisted laser desorption ionization (MALDI) mass spectrometric imaging. It mentions the issues faced while using chromatic lenses and also discusses the various properties of achromatic objective lens which makes it ideal for MALDI Mass Spectrometric Imaging, used for bio marker discovery and disease diagnosis in bio clinical research.

These are definitively exciting times for membrane lipid researchers. Once considered just as the cell membrane building blocks, the important role these lipids play is steadily being acknowledged. The improvement occurred in mass spectrometry techniques (MS) allows the establishment of the precise lipid composition of biological extracts. However, to fully understand the biological function of each individual lipid species, we need to know its spatial distribution and dynamics. In the past 10 years, the field has experienced a profound revolution thanks to the development of MS-based techniques allowing lipid imaging (MSI). Images reveal and verify what many lipid researchers had already shown by different means, but none as convincing as an image: each cell type presents a specific lipid composition, which is highly sensitive to its physiological and pathological state. While these techniques will help to place membrane lipids in the position they deserve, they also open the black box containing all the unknown regulatory mechanisms accounting for such tailored lipid composition. Thus, these results urges to different disciplines to redefine their paradigm of study by including the complexity revealed by the MSI techniques. [ABSTRACT FROM AUTHOR]

Arabinoxylans (AX) and (1→3),(1→4)-β-glucans (BG) are the major components of wheat grain cell walls. Although incompletely described at the molecular level, it is known that the chemical and distributional heterogeneity of these compounds impacts the quality and use of wheat. In this work, an emerging technique based on MALDI mass spectrometry imaging (MSI) was employed to map variations in the quantity, localization, and structure of these polysaccharides in the endosperm during wheat maturation. MALDI MSI couples detailed structural information with the spatial localization observed at the micrometer scale. The enzymic hydrolysis of AX and BG was performed directly on the grain sections, resulting in the efficient formation of smaller oligosaccharides that are easily measurable through MS, with no relocation across the grain. The relative quantification of the generated oligosaccharides was achieved. The method was validated by confirming data previously obtained using other analytical techniques. Furthermore, in situ analysis of grain cell walls through MSI revealed previously undetectable intense acetylation of AX in young compared to mature grains, together with findings concerning the feruloylation of AX and different structural features of BG. These results provide new insights into the physiological roles of these polysaccharides in cell walls and the specificity of the hydrolytic enzymes involved. [ABSTRACT FROM PUBLISHER]

Riverine tsetse ( Glossina) as Glossina palpalis gambiensis Vanderplank 1949 and Glossina tachinoides Westwood 1850 are the main vectors for African animal trypanosomoses in Burkina Faso. Vector control has been proven efficient in disease containment, but its success is endangered by the reinvasion of tsetse from neighbouring areas. Thus, identifying relic populations can enhance the success rate of vector control efforts. This is currently carried out through microsatellite analysis which is time-consuming and costly. Recently, matrix-assisted laser desorption/ionisation time-of-flight mass spectrometry-based analysis has become a routine method in microbial species identification. Owing to the rapidness and cost-effectiveness, this approach has been extended towards species identification of higher organisms such as tsetse. Following the recent experiences in distinguishing two genotypes of Prototheca spp., it is of interest to explore the validity of mass spectrometry for tsetse population differentiation. As a preliminary test, we submitted male and female G. palpalis gambiensis and G. tachinoides from Sideradougou and Folonzo, Burkina Faso (distance 60 km) to matrix-assisted laser desorption/ionisation analysis. The wing samples were utilized for protein extraction and mass spectra in a broad mass to charge ratio (2,000-20,000 kDa) were obtained. Specific peaks appeared to represent species, sex and location. Then, a peak list was extracted, containing the peaks in mass-to-charge ratio by revealing their intensities as well. These lists were used to compute a spectral dendrogram and a principle component analysis which displayed the differences among the samples from the two different regions. The results indicate that this technique can be extended with additional tsetse species, ideally with supporting genomic data, to later assist in designing rational vector control strategies. [ABSTRACT FROM AUTHOR]

Activation of sphingomyelinase (SMase) by extracellular stimuli is the major pathway for cellular production of ceramide, a bioactive lipid mediator acting through sphingomyelin (SM) hydrolysis. Previously, we reported the existence of six forms of neutral pH–optimum and Mg2+-dependent SMase (N-SMase) in the membrane fractions of bovine brain. Here, we focus on N-SMase ε from salt-extracted membranes. After extensive purification by 12,780-fold with a yield of 1.3%, this enzyme was eventually characterized as N-SMase2. The major single band of 60-kDa molecular mass in the active fractions of the final purification step was identified as heat shock protein 60 (Hsp60) by matrix-assisted laser desorption/ionization time-of-flight mass spectrometric analysis. Proximity ligation assay and immunoprecipitation study showed that Hsp60 interacted with N-SMase2, prompting us to examine the effect of Hsp60 on N-SMase2 and ceramide production. Interestingly, Hsp60 siRNA treatment significantly increased the protein level of N-SMase2 in N-SMase2-overexpressed HEK293 cells. Furthermore, transfection of Hsp60 siRNA into PC12 cells effectively increased both N-SMase activity and ceramide production and increased dopamine re-uptake with paralleled increase. Taken together, these results show that Hsp60 may serve as a negative regulator in N-SMase2-induced dopamine re-uptake by decreasing the protein level of N-SMase2. [ABSTRACT FROM AUTHOR]

In this study, we developed a platform that can be used to rapidly enrich polyhistidine(His)-tagged proteins/peptides from complex samples selectively using the Fe3O4@Al2O3 magnetic nanoparticles (MNPs) as the affinity probes. At pH 7, the dissociation constant between poly-His, i.e., His6, and the Fe3O4@Al2O3 MNPs was 10–5 M and the trapping capacity was 100 nmol/mg for His6. Enrichment was achieved by vigorously mixing the sample solution (<2 μL) and the MNPs (1–3 μg) by pipetting directly onto a matrix-assisted laser desorption/ionization (MALDI) plate for 10 s. The time for the enrichment and the sample volume required for analysis are therefore greatly reduced. After enrichment, the MNP–target species conjugates were promptly isolated by positioning a magnet on the edge of the sample well to aggregate the conjugates into a small spot within 5 s so that the nontarget species could be easily removed. Additionally, the problem of finding “sweet spots” on the target species during the MALDI mass spectrometry (MS) analysis was greatly reduced by magnetically isolating the target species on the MALDI plate. The limit of detection for His6 was, therefore, as low as 400 amol. His6 and AHHAHHAAD AHHAHHAAD spiked in a protein digest and in human plasma, respectively, were used as the samples to demonstrate the practicability of this approach in selective enrichment of His-rich peptides from complex samples. We also characterized His6-tagged proteins enriched on-plate by the Fe3O4@Al2O3 MNPs followed by on-plate tryptic digestion, selective enrichment, and MALDI-MS analysis. This approach can be used to determine quickly whether His6-tagged species are present in a sample. In addition, cell lysates containing recombinant Shiga-like toxins tagged with His6 were used as the samples to further demonstrate that the feasibility of this approach in analyzing very complex samples. The entire analysis process, including the on-plate enrichment and enzymatic digestion followed by MALDI-MS analysis, can be completed within 10 min. [ABSTRACT FROM AUTHOR]

A method is proposed for the analysis of O-glycans as 9-fluorenylmethyl (Fmoc) derivatives. After releasing the O-glycans from the protein backbone in the presence of ammonia-based media, the glycosylamines thus formed are conveniently labeled with Fmoc-Cl and analyzed by HPLC and MALDI-TOF MS after easy purification. Fmoc labeled O-glycans showed 3.5 times higher sensitivities than those labeled with 2-aminobenzoic acid in fluorescent detection. Various types of O-glycans having sialic acids, fucose, and/or sulfate residues were successfully labeled with Fmoc and analyzed by HPLC and MALDI-TOF MS. The method was applied to the comprehensive analysis of O-glycans expressed on MKN45 cells (human gastric adenocarcinoma). In addition, Fmoc-derivatized O-glycans were easily converted to free hemiacetal or glycosylamine-form glycans that are available for fabrication of glycan array and neoglycoproteins. To demonstrate the availability of our methods, we fabricate the glycan array with Fmoc labeled glycans derived from mucin samples and cancer cells. The model studies using the glycan array showed clear interactions between immobilized glycans and some lectins. [ABSTRACT FROM AUTHOR]

We have developed targets precoated with matrix for imaging lipids in tissues using matrix-assisted laser desorption ionization mass spectrometry (MALDI MS). Thin tissue sections (rat kidney and mouse or rat brains) were placed onto 1,5-diaminonaphthalene precoated targets (prepared beforehand by a protocol utilizing sublimation) and were washed with ammonium formate solution. After a brief drying period, the target slides were imaged by MALDI MS. The resulting images from these sections were of equivalent quality to those obtained using the usual postcoating approach, such as sublimation and spraying, in terms of the sharpness of substructures in the images demonstrated by imaging at spatial resolutions of 100, 10, and 5 μm. Matrix precoated targets have a shelf life of more than 6 months when kept in a dark, nonhumid environment such as a nontransparent desiccator. [ABSTRACT FROM AUTHOR]

Mass spectrometric imaging (MSI) is an analytical technique used to determine the distribution of individual analytes within a given sample. A wide array of analytes and samples can be investigated by MSI, induding drug distribution in rats, lipid analysis from brain tissue, protein differentiation in tumors, and plant metabolite distributions. Matrix-assisted laser desorption/ionization (MALDI) is a soft ionization technique capable of desorbing and ionizing a large range of compounds, and it is the most common ionization source used in MSL MALDI mass spectrornetry (MS) is generally considered to be a qualitative analytical technique because of significant ion-signal variability. Consequently, MSI is also thought to be a qualitative technique because of the quantitative limitations of MALDI coupled with the homogeneity of tissue sections inherent in an MSI experiment. Thus, condusions based on MS images are often limited by the inability to correlate ion signal increases with actual concentration increases. Here, we report a quantitative MSI method for the analysis of cocaine (COC) from brain tissue using a deuterated internal standard (COC-d3) combined with wide-isolation MS/MS for analysis of the tissue extracts with scan-by-scan COC-to-COC-d3 normalization. This resulted in significant improvements in signal reproducibility and calibration curve linearity. Quantitative results from the MSI experiments were compared with quantitative results from liquid chromatography (LC)-MS/MS results from brain tissue extracts. Two different quantitative MSI techniques (standard addition and external calibration) produced quantitative results comparable to LC-MS/MS data. Tissue extracts were also analysed by MALDI wide- isolation MS/MS, and quantitative results were nearly identical to those from LC-MS/MS. These results dearly demonstrate the necessity for an internal standard for quantitative MSI experiments. [ABSTRACT FROM AUTHOR]

The review explains the fundamentals of the methods of mass spectrometry with matrix assisted laser desorption/ionization (MALDI) and mass spectrometry with surface-assisted laser desorption/ionization (SALDI). Basics of these mass spectrometric techniques and regularities of applications of matrixes of various types are discussed. Examples of using the MALDI and SALDI techniques are presented for obtaining spectra of low-molecular compounds and compounds with a high molecular mass in connection with studies of corrosion inhibitors. [ABSTRACT FROM AUTHOR]

The article focuses on a study regarding the impact of second matrix layer on matrix-assisted laser desorption-ionization (MALDI) time-of-flight (TOF) mass spectrometry unclassified target spots that suggests that mass spectra quality was improved by the layer.

The author reflects on a clinical approach to antibiotic treatment presented in a study that is antimicrobial de-escalation (ADE) to limit unnecessary antimicrobial exposure for reducing antibiotic resistance. He states that there is no evidence of lower emergence of resistance resulted by ADE. The author suggests that the impact of matrix-assisted laser desorption-ionization (MALDI-TOF) mass spectrometry (MS) can be analyzed in patients with bloodstream infections.

The article discusses a study to identify of two genes associated with Generalized Pustular Psoriasis (GPP) approved by the Ethics Committee of Fudan University in Shanghai, China. Topics discussed include identifying single-nucleotide polymorphisms of the linkage disequilibrium (LD) region using matrix-assisted laser desorption/ionization, analyzing mass spectrograms with MassARRAY TYPER software and detecting allelic by employing the Armitage trend test using Plink 1.07 software.

The authors comment on the matrix-assisted laser desorption/ionization time-of-flight mass spectrometry profiling (MALDI-TOF MS profiling) approach used for the accurate identification of microorganism species in the field of proteomics. Topics include the advances and applications of the approach, the use of MALDI-TOF MS profiling in plant proteomics, and the challenge of identifying the proteins detected in the MALDI-TOF MS profiles.

In bacteriophages related to T4, hydroxymethylcytosine (hmC) is incorporated into the genomic DNA during DNA replication and is then further modified to glucosyl-hmC by phage-encoded glucosyltransferases. Previous studies have shown that RB69 shares a core set of genes with T4 and relatives. However, unlike the other “RB” phages, RB69 is unable to recombine its DNA with T4 or with the other “RB” isolates. In addition, despite having homologs to the T4 enzymes used to synthesize hmC, RB69 has no identified homolog to known glucosyltransferase genes. In this study we sought to understand the basis for RB69’s behavior using high-pH anion exchange chromatography (HPAEC) and mass spectrometry. Our analyses identified a novel phage epigenetic DNA sugar modification in RB69 DNA, which we have designated arabinosyl-hmC (ara-hmC). We sought a putative glucosyltranserase responsible for this novel modification and determined that RB69 also has a novel transferase gene, ORF003c, that is likely responsible for the arabinosyl-specific modification. We propose that ara-hmC was responsible for RB69 being unable to participate in genetic exchange with other hmC-containing T-even phages, and for its described incipient speciation. The RB69 ara-hmC also likely protects its DNA from some anti-phage type-IV restriction endonucleases. Several T4-related phages, such as E. coli phage JS09 and Shigella phage Shf125875 have homologs to RB69 ORF003c, suggesting the ara-hmC modification may be relatively common in T4-related phages, highlighting the importance of further work to understand the role of this modification and the biochemical pathway responsible for its production. [ABSTRACT FROM AUTHOR]