Immunoreactive Proteins in the Esophageal Gland Cells of Anisakis Simplex Sensu Stricto Detected by MALDI-TOF/TOF Analysis

In plant and animal nematode parasites, proteins derived from esophageal gland cells have been shown to be important in the host-nematodes relationship but little is known about the allergenic potential of these proteins in the genus Anisakis. Taking into account the increase of anisakiasis and allergies related to these nematodes, immunoreactive properties of gland cell proteins were investigated. Two hundred ventricles were manually dissected from L3 stage larvae of Aniskakis simplex s.s. to allow direct protein analysis. Denaturing gel electrophoresis followed by monochromatic silver staining which revealed the presence of differential (enriched) proteins when compared to total nematode extracts. Such comparison was performed by means of 1D and 2D electrophoresis. Pooled antisera from Anisakis spp.-allergic patients were used in western blots revealing the presence of 13 immunoreactive bands in the ventricular extracts in 1D, with 82 spots revealed in 2D. The corresponding protein bands and spots were excised from the silver-stained gel and protein assignation was made by MALDI-TOF/TOF. A total of 13 (including proteoforms) were unambiguously identified. The majority of these proteins are known to be secreted by nematodes into the external environment, of which three are described as being major allergens in other organisms with different phylogenetic origin and one is an Anisakis simplex allergen.

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In Planta Localization of a β-1,4-Endoglucanase Secreted by Heterodera glycines

Molecular Plant-Microbe Interactions ◽

10.1094/mpmi.1999.12.1.64 ◽

1999 ◽

Vol 12 (1) ◽

pp. 64-67 ◽

Cited By ~ 85

Author(s):

Xiaohong Wang ◽

Diane Meyers ◽

Yitang Yan ◽

Thomas Baum ◽

Geert Smant ◽

...

Keyword(s):

Cross Sections ◽

Soybean Cyst Nematode ◽

Heterodera Glycines ◽

In Planta ◽

Root Cortex ◽

Western Blots ◽

Soybean Root ◽

Second Stage ◽

Gland Cells ◽

Esophageal Gland

Polyclonal sera specific to β-1,4-endoglucanases (cellulases) synthesized in the subventral esophageal gland cells of the soybean cyst nematode, Heterodera glycines, were used to provide the first identification of a nematode esophageal gland protein that is secreted into host plant tissue. Sera generated to proteins encoded by Hg-eng-1 and Hg-eng-2 (endoglucanases) did not cross-react with soybean root proteins on Western blots (immunoblots) or in immunofluorescence microscopy of noninoculated (control) soybean root sections. In cross sections of soybean roots at 24 h after inoculation of roots with second-stage juveniles of H. glycines, HG-ENG-1 was localized within the nematode's subventral gland cells and was not detected in root tissue. HG-ENG-2 was localized within the subventral gland cells and was secreted from the juvenile's cortical tissue at 24 h after inoculation of roots with second-stage juveniles of H. glycines. HG-ENG-2 was localized along the juvenile's migratory path through the root cortex.

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A case series of medically managed Candida parapsilosis complex prosthetic valve endocarditis

Annals of Clinical Microbiology and Antimicrobials ◽

10.1186/s12941-020-00409-4 ◽

2021 ◽

Vol 20 (1) ◽

Author(s):

Penghao Guo ◽

Yuting He ◽

Rui Fan ◽

Zhongwen Wu ◽

Yili Chen ◽

...

Keyword(s):

Prosthetic Valve ◽

Candida Parapsilosis ◽

Case Series ◽

Prosthetic Valve Endocarditis ◽

Sensu Stricto ◽

Maldi Tof Ms ◽

Maldi Tof ◽

Candida Metapsilosis ◽

Candida Orthopsilosis ◽

Tof Ms

Abstract Background In recent years, Candida parapsilosis is recognized as a species complex and is composed of Candida parapsilosis sensu stricto, Candida orthopsilosis and Candida metapsilosis. Candida parapsilosis complex prosthetic valve endocarditis (PVE) is rare and the survival rate is still low despite of optimal therapeutic strategies. In our report, it is novel to report cases as Candida parapsilosis complex PVE at species and identify Candida parapsilosis using MALDI-TOF MS. Case presentation A series of 4 cases of Candida parapsilosis complex PVE from our institution was reported. Three were infected by Candida parapsilosis sensu stricto and one was infected by Candida metapsilosis. The condition of two cases got better and the other died. Conclusions More attention should be paid to Candida parapsilosis complex PVE and early diagnosis and prompt antibiotic therapy may play a role in the treatment for Candida parapsilosis complex PVE. It is recommended to identify Candida parapsilosis complex at species level and MALDI-TOF MS as an easy, fast and efficient identification method is worth promoting in clinical microbiology

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Microaspiration of Esophageal Gland Cells and cDNA Library Construction for Identifying Parasitism Genes of Plant-Parasitic Nematodes

Methods in Molecular Biology - Plant Immunity ◽

10.1007/978-1-61737-998-7_9 ◽

2011 ◽

pp. 89-107 ◽

Cited By ~ 6

Author(s):

Richard S. Hussey ◽

Guozhong Huang ◽

Rex Allen

Keyword(s):

Cdna Library ◽

Parasitic Nematodes ◽

Plant Parasitic Nematodes ◽

Cdna Library Construction ◽

Library Construction ◽

Gland Cells ◽

Esophageal Gland ◽

Parasitism Genes ◽

Plant Parasitic

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The Selection of Reliable Reference Genes for RT-qPCR Analysis of Anisakis simplex Sensu Stricto Gene Expression from Different Developmental Stages

Acta Parasitologica ◽

10.2478/s11686-020-00220-3 ◽

2020 ◽

Vol 65 (4) ◽

pp. 837-842 ◽

Cited By ~ 1

Author(s):

Elżbieta Łopieńska-Biernat ◽

Robert Stryiński ◽

Łukasz Paukszto ◽

Jan Paweł Jastrzębski ◽

Karol Makowczenko

Keyword(s):

Gene Expression ◽

Reference Genes ◽

Parasitic Nematode ◽

Developmental Stages ◽

Receptor Gene ◽

Elongation Factor ◽

Sensu Stricto ◽

Nuclear Hormone Receptor ◽

Anisakis Simplex ◽

Complex Life Cycle

Abstract Background Anisakis simplex s. s. is a parasitic nematode with a complex life cycle in which humans can become accidental hosts by consuming raw or not fully cooked fish containing L3 larvae. The growing popularity of raw fish dishes has contributed to an increase in the incidence of anisakiasis, which has spurred scientific efforts to develop new methods for diagnosing and treating the disease and also to investigate the gene expression at different developmental stages of this parasite. The identification of reference genes suitable for the normalization of RT-qPCR data has not been studied with respect to A. simplex s. s. Methods In the present study, eight candidate reference genes were analyzed in A. simplex s. s. at two different developmental stages: L3 and L4. The expression stability of these genes was assessed by geNorm and NormFinder softwares. Results In general, our results identified translation elongation factor 1α (ef-1α) and peptidyl-prolyl isomerase 12 (ppi12) as the most stable genes in L3 and L4 developmental stages of A. simplex s. s. Validation of the selected reference genes was performed by profiling the expression of the nuclear hormone receptor gene (nhr 48) in different developmental stages. Conclusions This first analysis selecting suitable reference genes for RT-qPCR in A. simplex s. s. will facilitate future functional analyses and deep mining of genetic resources in this parasitic nematode.

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Comparative Transcriptomics Reveals Clues for Differences in Pathogenicity between Hysterothylacium aduncum, Anisakis simplex sensu stricto and Anisakis pegreffii

Genes ◽

10.3390/genes11030321 ◽

2020 ◽

Vol 11 (3) ◽

pp. 321 ◽

Cited By ~ 4

Author(s):

Serena Cavallero ◽

Fabrizio Lombardo ◽

Marco Salvemini ◽

Antonella Pizzarelli ◽

Cinzia Cantacessi ◽

...

Keyword(s):

Ribosomal Proteins ◽

De Novo ◽

Proteolytic Enzymes ◽

Sensu Stricto ◽

Anisakis Simplex ◽

Anisakis Pegreffii ◽

Primary Hemostasis ◽

Differential Gene ◽

Immunomodulatory Peptides ◽

Hysterothylacium Aduncum

Ascaridoid nematodes are widespread in marine fishes. Despite their major socioeconomic importance, mechanisms associated to the fish-borne zoonotic disease anisakiasis are still obscure. RNA-Seq and de-novo assembly were herein applied to RNA extracted from larvae and dissected pharynx of Hysterothylacium aduncum (HA), a non-pathogenic nematode. Assembled transcripts in HA were annotated and compared to the transcriptomes of the zoonotic species Anisakis simplex sensu stricto (AS) and Anisakis pegreffii (AP). Approximately 60,000,000 single-end reads were generated for HA, AS and AP. Transcripts in HA encoded for 30,254 putative peptides while AS and AP encoded for 20,574 and 20,840 putative peptides, respectively. Differential gene expression analyses yielded 471, 612 and 526 transcripts up regulated in the pharynx of HA, AS and AP. The transcriptomes of larvae and pharynx of HA were enriched in transcripts encoding collagen, peptidases, ribosomal proteins and in heat-shock motifs. Transcripts encoding proteolytic enzymes, anesthetics, inhibitors of primary hemostasis and virulence factors, anticoagulants and immunomodulatory peptides were up-regulated in AS and AP pharynx. This study represents the first transcriptomic characterization of a marine parasitic nematode commonly recovered in fish and probably of negligible concern for public health.

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Influence of Instant Controlled Pressure Drop (DIC) on Allergenic Potential of Tree Nuts

Molecules ◽

10.3390/molecules25071742 ◽

2020 ◽

Vol 25 (7) ◽

pp. 1742 ◽

Cited By ~ 2

Author(s):

Fatima Vicente ◽

Africa Sanchiz ◽

Rosa Rodríguez-Pérez ◽

Maria Pedrosa ◽

Santiago Quirce ◽

...

Keyword(s):

Pressure Drop ◽

Conformational Changes ◽

Food Processing ◽

Electrophoretic Pattern ◽

Temperature Treatment ◽

Western Blots ◽

Allergenic Potential ◽

Allergenic Proteins ◽

Instant Controlled Pressure Drop ◽

Suitable Technique

Pistachio and cashew contain allergenic proteins, which causes them to be removed from the diet of allergic people. Previous studies have demonstrated that food processing (thermal and non-thermal) can produce structural and/or conformational changes in proteins by altering their allergenic capacity. In this study, the influence of instant controlled pressure drop (DIC) on pistachio and cashew allergenic capacity has been studied. Western blot was carried out using IgG anti-11S and anti-2S and IgE antibodies from sera of patients sensitized to pistachio and cashew. DIC processing causes changes in the electrophoretic pattern, reducing the number and intensity of protein bands, as the pressure and temperature treatment increment, which results in a remarkable decrease in detection of potentially allergenic proteins. The harshest conditions of DIC (7 bar, 120 s) markedly reduce the immunodetection of allergenic proteins, not only by using IgG (anti 11S and anti 2S) but also when IgE sera from sensitized patients were used for Western blots. Such immunodetection is more affected in pistachio than in cashew nuts, but is not completely removed. Therefore, cashew proteins are possibly more resistant than pistachio proteins. According these findings, instant controlled pressure drop (DIC) can be considered a suitable technique in order to obtain hypoallergenic tree nut flour to be used in the food industry.

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Rapid Detection of Echinocandins Resistance by MALDI-TOF MS in Candida parapsilosis Complex

Microorganisms ◽

10.3390/microorganisms8010109 ◽

2020 ◽

Vol 8 (1) ◽

pp. 109

Author(s):

Ana Emília M. Roberto ◽

Danilo E. Xavier ◽

Esteban E. Vidal ◽

Cláudia Fernanda de L. Vidal ◽

Rejane P. Neves ◽

...

Keyword(s):

Mass Spectrometry ◽

Incubation Time ◽

Candida Parapsilosis ◽

Clinical Laboratory ◽

Antifungal Susceptibility ◽

Reference Method ◽

Sensu Stricto ◽

Maldi Tof ◽

Separation Parameter ◽

Candida Parapsilosis Complex

Mass spectrometry by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) was used to identify and differentiate the pattern of susceptibility of clinical isolates of Candida parapsilosis complex. 17 C. parapsilosis sensu stricto, 2 C. orthopsilosis, and 1 C. metapsilosis strains were obtained from blood cultures, and three different inocula (103, 105, and 107 CFU/mL) were evaluated against three echinocandins at concentrations ranging from 0.03 to 16 µg/mL after incubation of 1 h, 2 h, and 3 h. Drug-free control was used. The spectra obtained at these concentrations were applied to generate composite correlation index (CCI) matrices for each yeast individually. After cross correlations and autocorrelations of each spectra with null (zero) and maximal (16) concentrations, the CCI was used as separation parameter among spectra. Incubation time and inoculum were critical factors to reach higher precision and reliability of this trial. With an incubation time of 3 h and inoculum of 107 CFU/mL, it was possible to determine the breakpoint of the clinical yeasts by MALDI-TOF that presented high agreement with the clinical laboratory standard institute (CLSI) reference method. Herein, we show that mass spectrometry using the MALDI-TOF technique is powerful when it exploits antifungal susceptibility testing assays.

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