scholarly journals Identification of a Spiroplasma citri hydrophilic protein associated with insect transmissibility

Microbiology ◽  
2006 ◽  
Vol 152 (4) ◽  
pp. 1221-1230 ◽  
Author(s):  
Nabil Killiny ◽  
Brigitte Batailler ◽  
Xavier Foissac ◽  
Colette Saillard
Keyword(s):  
Close Contact ◽  
Genome Project ◽  
Insect Vector ◽  
Electron Microscopic ◽  
Protein Database ◽  
Two Dimensional Gel Electrophoresis ◽  
Ph Gradients ◽  
Spiroplasma Citri ◽  
Flight Mass Spectrometry ◽  
Pcr Products

With the aim of identifying Spiroplasma citri proteins involved in transmission by the leafhopper Circulifer haematoceps, protein maps of four transmissible and four non-transmissible strains were compared. Total cell lysates of strains were analysed by two-dimensional gel electrophoresis using commercially available immobilized pH gradients (IPGs) covering a pH range of 4–7. Approximately 530 protein spots were visualized by silver staining and the resulting protein spot patterns for the eight strains were found to be highly similar. However, comparison using PDQuest 2-D analysis software revealed two trains of protein spots that were present only in the four transmissible strains. Using MALDI-TOF (matrix-assisted laser desorption/ionization time-of-flight) mass spectrometry and a nearly complete S. citri protein database, established during the still-ongoing S. citri GII-3-3X genome project, the sequences of both proteins were deduced. One of these proteins was identified in the general databases as adhesion-related protein (P89) involved in the attachment of S. citri to gut cells of the insect vector. The second protein, with an apparent molecular mass of 32 kDa deduced from the electrophoretic mobility, could not be assigned to a known protein and was named P32. The P32-encoding gene (714 bp) was carried by a large plasmid of 35·3 kbp present in transmissible strains and missing in non-transmissible strains. PCR products with primers designed from the p32 gene were obtained only with genomic DNA isolated from transmissible strains. Therefore, P32 has a putative role in the transmission process and it could be considered as a marker for S. citri leafhopper transmissibility. Functional complementation of a non-transmissible strain with the p32 gene did not restore the transmissible phenotype, despite the expression of P32 in the complemented strain. Electron microscopic observations of salivary glands of leafhoppers infected with the complemented strain revealed a close contact between spiroplasmas and the plasmalemma of the insect cells. This further suggests that P32 protein contributes to the association of S. citri with host membranes.

A scanning electron microscopic study of the eggs of Culicoides variipennis (Diptera: Ceratopogonidae)

1987 ◽  
Vol 45 ◽  
pp. 884-885 ◽  
Author(s):  
R. A. Nunamaker ◽  
C. E. Nunamaker ◽  
B. C. Wick
Keyword(s):  
Distinct Species ◽  
Hemorrhagic Disease ◽  
Insect Vector ◽  
Electron Microscopic ◽  
Important Species ◽  
Scanning Electron Microscopic Study ◽  
Laboratory Colony ◽  
Culicoides Variipennis ◽  
Morphological Differences ◽  
Scanning Electron

Culicoides variipennis (Coquillett) is probably the most economically important species of biting midge in the U.S. due to its involvement in the transmission of bluetongue (BT) disease of sheep, cattle and ruminant wildlife, and epizootic hemorrhagic disease (EHD) of deer. Proposals have been made to recognize the eastern and western populations of this insect vector as distinct species. Others recommend use of the term “variipennis complex” until such time that the necessary biosystematic studies have been made to determine the genetic nature and/or minute morphological differences within the population structure over the entire geographic range of the species. Increasingly, students of ootaxonomy are relying on scanning electron microscopy (SEM) to assess chorionic features. This study was undertaken to provide comparative chorionic data for the C. variipennis complex.Culicoides variipennis eggs were collected from a laboratory colony maintained in Laramie, Wyoming.

Identification of Sulfur Activation Relevant Protein Genes of Extremely Thermophilic Acidianus manzaensis

2017 ◽  
Vol 262 ◽  
pp. 461-465 ◽  
Author(s):  
Hong Chang Liu ◽  
Jin Lan Xia ◽  
Zhen Yuan Nie ◽  
Ya Long Ma ◽  
Yun Yang ◽  
...  
Keyword(s):  
Gel Electrophoresis ◽  
Elemental Sulfur ◽  
Extracellular Proteins ◽  
Laser Desorption Ionization ◽  
Two Dimensional Gel Electrophoresis ◽  
Ionization Time ◽  
Real Time Quantitative Pcr ◽  
Flight Mass Spectrometry ◽  
Genes Encoding ◽  
Acidianus Manzaensis

The sulfur activation by extracellular proteins is considered as the crucial stage during biooxidation of elemental sulfur (S0). In order to study genes encoding sulfur-activation related extracellular proteins of extremely thermophilic Acidianus manzaensis, the extracellular proteins with higher abundance for the strain grown on S0 allotropes than that on Fe2+ were first screened by two-dimensional gel electrophoresis, and then identified by matrix-assisted laser desorption ionization time-of-flight mass spectrometry. Nine genes amplified with PCR were satisfactory according to their agarose gel electrophoresis. The differential expression of these nine genes when the strain grown on S0 allotropes and Fe2+ were analyzed with real-time quantitative PCR (RT-qPCR). Results showed that seven of them were higher expressed when the strain grown on S0 allotropes than on Fe2+, indicating they may be related with sulfur activation by A. manzaensis.

scholarly journals First Report of an Aster Yellows Phytoplasma Associated with Cabbage in Southern Texas

Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 447-447 ◽  
Author(s):  
I.-M. Lee ◽  
R. A. Dane ◽  
M. C. Black ◽  
Noel Troxclair
Keyword(s):  
16S Rdna ◽  
Nested Pcr ◽  
Restriction Enzymes ◽  
Diagnostic Procedures ◽  
Acid Extraction ◽  
Insect Vector ◽  
Aster Yellows ◽  
First Report ◽  
Pcr Products ◽  
Aster Yellows Phytoplasma

In early spring 2000 carrot crops in southwestern Texas were severely infected by an outbreak of phyllody associated with aster yellows phytoplasma. Cabbage crops that had been planted adjacent to these carrot fields began to display previously unobserved symptoms characteristic of phytoplasma infection. Symptoms included purple discoloration in leaf veins and at the outer edges of leaves on cabbage heads. Proliferation of sprouts also occurred at the base of the stem and between leaf layers of some plants, and sprouts sometimes continued to proliferate on extended stems. About 5% of cabbage plants in the field exhibited these symptoms. Two symptomless and four symptomatic cabbage heads were collected in early April from one cabbage field. Veinal tissues were stripped from each sample and used for total nucleic acid extraction. To obtain specific and sufficient amount of PCR products for analysis, nested PCR was performed by using primer pairs (first with P1/P7 followed by R16F2n/R16R2) (1,2) universal for phytoplasma detection. A specific 16S rDNA fragment (about 1.2 kb) was strongly amplified from the four symptomatic but not from the two asymptomatic samples. The nested PCR products obtained from the four symptomatic samples were then analyzed by restriction fragment length polymorphism (RFLP) using the restriction enzymes MseI, HhaI, and HpaII, and the RFLP patterns were compared to the published patterns of known phytoplasmas (1). The resulting RFLP patterns were identical to those of a phytoplasma belonging to subgroup B of the aster yellows phytoplasma group (16SrI). These RFLP patterns were also evident in putative restriction sites observed in a 1.5 kbp nucleotide sequence of the 16S rDNA. This is the first report of aster yellows phytoplasma associated disease symptoms in cabbage in Texas. The occurrence of cabbage proliferation coincided with the presence of high populations of the insect vector, aster leafhopper. References: (1) I.-M. Lee et al. Int. J. Syst. Bacteriol. 48:1153, 1998. (2) B. Schneider et al. 1995. Molecular and Diagnostic Procedures in Mycoplasmology, Vol. I. Academic Press, San Diego, CA.

scholarly journals Preliminary study of coronavirus disease 2019 on pets in pandemic in Denpasar, Bali, Indonesia

2021 ◽  
pp. 2979-2983
Author(s):  
Hamong Suharsono ◽  
Ali Ghufron Mukti ◽  
Ketut Suryana ◽  
I. Wayan Masa Tenaya ◽  
Dilasdita Kartika Pradana ◽  
...  
Keyword(s):  
Close Contact ◽  
Genetic Material ◽  
Rna Isolation ◽  
World Health ◽  
Intermediate Hosts ◽  
Rt Pcr ◽  
Pcr Products ◽  
Preliminary Study ◽  
Health Organization ◽  
Respiratory Droplets

Background and Aim: Coronavirus disease 2019 (COVID-19) is an acute infectious respiratory disease caused by severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) and has spread rapidly globally, resulting in a pandemic. In humans, the main routes of transmission are respiratory droplets and close contact with infected individuals or through contact with an object infected with the virus, followed by touching mouth, nose, or eyes. It is assumed that SARS-CoV-2 was originated in wild animals and was then transmitted to humans. Although some wildlife and domestic animals can be naturally or experimentally infected with the virus, the intermediate hosts that transmitted it to humans are still unknown. Understanding the dynamics of SARS-CoV-2 associated with possible zoonotic transmission of intermediate hosts is considered critical. Reportedly, cats or dogs living with COVID-19-positive humans tested positive for the disease, suggesting that the virus was transmitted to the animals from humans. Information regarding the epidemiological investigation and comprehensive studies is limited. Therefore, it is still unclear how high is the correlation of infection in humans and pet animals, especially those living together. The aim of this study was to investigate the possibility of SARS-CoV-2 infection in the pets of patients with COVID-19 who were hospitalized at the Wangaya hospital, Denpasar, Bali, Indonesia. Materials and Methods: A total of seven clinically asymptomatic pets (six dogs of different races and sexes and a cat [age, 360-2920 days]) were included in this study. These animals belonged to patients with confirmed SARS-CoV-2 infection from August to November 2020. Nasal swab and nasopharyngeal samples were collected from the pets individually under anesthetic condition and were collected 6-12 days after confirmed SARS-CoV-2 infection in owners and hospitalization at the Wangaya Hospital. The swab samples were then processed for RNA isolation and tested using reverse transcription-polymerase chain reaction (RT-PCR) for SARS-CoV-2, in accordance with the World Health Organization manual 2020. Results: RT-PCR results for all seven RNA samples, prepared from the swab samples, were negative. For the samples, all PCR products were below the threshold limit, suggesting no genetic material belonging to the samples tested. Conclusion: This was the first preliminary study of COVID-19 on pets in pandemic using RT-PCR. The study tested a very limited quantity of samples, and all of them were negative. However, the way in which the samples were prepared was considered appropriate. Therefore, in further studies, testing of more samples of pets of more individuals with confirmed SARS-CoV-2 infection is required.

scholarly journals Host cell adhesion to Schistosoma mansoni larvae in the peritoneal cavity of naive mice: histological and scanning electron microscopic studies

1993 ◽  
Vol 35 (1) ◽  
pp. 17-22 ◽  
Author(s):  
Alan Lane de Melo ◽  
Conceição Ribeiro da Silva Machado ◽  
Leógenes Horácio Pereira
Keyword(s):  
Cell Adhesion ◽  
Schistosoma Mansoni ◽  
Host Cell ◽  
Peritoneal Cavity ◽  
Mononuclear Cells ◽  
Close Contact ◽  
Histological Study ◽  
Electron Microscopic ◽  
Microscopic Studies ◽  
Scanning Electron

Cercariae of Schistosoma mansoni inoculated into the peritoneal cavity of naive mice induced host cell adhesion to their surface, but after 90 minutes the number of adherent cells sharply decreased. The cell detachment is progressive and simultaneous to the cercaria-schistosomule transformation. The histological study showed mainly neutrophils in close contact with the larvae. Mononuclear cells and some eosinophils were occasionally seen surrounding the adherent neutrophils. The scanning electron microscopy showed cells displaying twisted microvilli and several microplicae contacting or spreading over the larval surface, and larvae completely surrounded by clusters of cells. These results suggest that the neutrophils recognize molecules on the cercarial surface which induce their spreading

scholarly journals Synovial Fluid Cell Proteomic Analysis Identifies Upregulation of Alpha-Taxilin Proteins in Rheumatoid Arthritis: A Potential Prognostic Marker

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Ashish Sarkar ◽  
Shivani Sharma ◽  
Prachi Agnihotri ◽  
Tanmoy Sarkar ◽  
Pooja Kumari ◽  
...  
Keyword(s):  
Rheumatoid Arthritis ◽  
Synovial Fluid ◽  
Prognostic Marker ◽  
Surrounding Tissue ◽  
Enzyme Linked Immunosorbent Assay ◽  
Two Dimensional Gel Electrophoresis ◽  
Flight Mass Spectrometry ◽  
Tissue Identification ◽  
Proteomic Approach ◽  
In Silico Studies

Rheumatoid arthritis (RA) is a chronic autoimmune inflammatory disease affecting the joints and surrounding tissue. Identification of novel proteins associated with the progression of a disease is a prerequisite for understanding the pathogenesis of RA. The present study was undertaken to identify the potential biomarkers from a less explored biological sample such as synovial fluid (SF) cells which is specific for RA and to analyze their functional aspects using proteomic approach. Two-dimensional gel electrophoresis (2-DE) was performed using synovial fluid cells of RA and osteoarthritis (OA) patients, and 7 differentially expressed proteins were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS/MS). Αlpha-Taxilin (α-Taxilin) has been found as one of the novel, significantly up regulated protein in RA. It has been validated in the synovium, synovial fluid (SF), SF cells, and plasma samples by Western blot, enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting (FACS), immunohistochemistry (IHC), and real-time PCR. The identification of autoantibody against α-Taxilin and in silico studies has further helped us to understand its involvement in disease mechanism. The present study will therefore provide knowledge towards the etiology of RA that pave the way for suitable prognostic marker identification along with other clinical parameters.

scholarly journals Comparative Proteomic Analysis of Extracellular Proteins of Enterohemorrhagic and Enteropathogenic Escherichia coli Strains and Their ihf and ler Mutants

2004 ◽  
Vol 70 (9) ◽  
pp. 5274-5282 ◽  
Author(s):  
M. Li ◽  
I. Rosenshine ◽  
S. L. Tung ◽  
X. H. Wang ◽  
D. Friedberg ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Polyacrylamide Gel Electrophoresis ◽  
Integration Host Factor ◽  
Extracellular Proteins ◽  
Major Protein ◽  
Human Pathogens ◽  
Protein Database ◽  
Flight Mass Spectrometry ◽  
Proteomic Approach ◽  
In The Wild

ABSTRACT Enterohemorrhagic and enteropathogenic Escherichia coli (EHEC and EPEC, respectively) strains are closely related human pathogens that are responsible for food-borne epidemics in many countries. Integration host factor (IHF) and the locus of enterocyte effacement-encoded regulator (Ler) are needed for the expression of virulence genes in EHEC and EPEC, including the elicitation of actin rearrangements for attaching and effacing lesions. We applied a proteomic approach, using two-dimensional polyacrylamide gel electrophoresis in combination with matrix-assisted laser desorption ionization-time of flight mass spectrometry and a protein database search, to analyze the extracellular protein profiles of EHEC EDL933, EPEC E2348/69, and their ihf and ler mutants. Fifty-nine major protein spots from the extracellular proteomes were identified, including six proteins of unknown function. Twenty-six of them were conserved between EHEC EDL933 and EPEC E2348/69, while some of them were strain-specific proteins. Four common extracellular proteins (EspA, EspB, EspD, and Tir) were regulated by both IHF and Ler in EHEC EDL933 and EPEC E2348/69. TagA in EHEC EDL933 and EspC and EspF in EPEC E2348/69 were present in the wild-type strains but absent from their respective ler and ihf mutants, while FliC was overexpressed in the ihf mutant of EPEC E2348/69. Two dominant forms of EspB were found in EHEC EDL933 and EPEC E2348/69, but the significance of this is unknown. These results show that proteomics is a powerful platform technology for accelerating the understanding of EPEC and EHEC pathogenesis and identifying markers for laboratory diagnoses of these pathogens.

scholarly journals Influence of Acute High Glucose on Protein Abundance Changes in Murine Glomerular Mesangial Cells

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Michelle T. Barati ◽  
James C. Gould ◽  
Sarah A. Salyer ◽  
Susan Isaacs ◽  
Daniel W. Wilkey ◽  
...  
Keyword(s):  
Protein Expression ◽  
Posttranslational Modifications ◽  
High Glucose ◽  
Mesangial Cells ◽  
Expression Patterns ◽  
Growth Conditions ◽  
Glomerular Mesangial Cells ◽  
Two Dimensional Gel Electrophoresis ◽  
Glucose Levels ◽  
Flight Mass Spectrometry

The effects of acute exposure to high glucose levels as experienced by glomerular mesangial cells in postprandial conditions and states such as in prediabetes were investigated using proteomic methods. Two-dimensional gel electrophoresis and matrix assisted laser desorption ionization time of flight mass spectrometry methods were used to identify protein expression patterns in immortalized rat mesangial cells altered by 2 h high glucose (HG) growth conditions as compared to isoosmotic/normal glucose control (NG⁎) conditions. Unique protein expression changes at 2 h HG treatment were measured for 51 protein spots. These proteins could be broadly grouped into two categories: (1) proteins involved in cell survival/cell signaling and (2) proteins involved in stress response. Immunoblot experiments for a protein belonging to both categories, prohibitin (PHB), supported a trend for increased total expression as well as significant increases in an acidic PHB isoform. Additional studies confirmed the regulation of proteasomal subunit alpha-type 2 and the endoplasmic reticulum chaperone and oxidoreductase PDI (protein disulfide isomerase), suggesting altered ER protein folding capacity and proteasomal function in response to acute HG. We conclude that short term high glucose induces subtle changes in protein abundances suggesting posttranslational modifications and regulation of pathways involved in proteostasis.

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