Isolation of transcripts overexpressed in the human pathogenTrichophyton rubrumgrown in lipid as carbon source

2011 ◽  
Vol 57 (4) ◽  
pp. 333-338 ◽  
Author(s):  
Fernanda C.A. Maranhão ◽  
Henrique C.S. Silveira ◽  
Antonio Rossi ◽  
Nilce M. Martinez-Rossi
Keyword(s):  
Carbon Source ◽  
Trichophyton Rubrum ◽  
Cdna Array ◽  
Subtractive Hybridization ◽  
Cellular Responses ◽  
Cdna Clones ◽  
Medical Importance ◽  
Insight Into ◽  
Dot Blotting ◽  
Host Invasion

Trichophyton rubrum is the most common etiological agent of human dermatophytosis. Despite the incidence and medical importance of this dermatophyte, little is known about the mechanisms of host invasion and pathogenicity. Host invasion depends on the adaptive cellular responses of the pathogen that allow it to penetrate the skin layers, which are mainly composed of proteins and lipids. In this study, we used suppression subtractive hybridization to identify transcripts overexpressed in T. rubrum cultured in lipid as carbon source. Among the subtractive cDNA clones isolated, 85 clones were positively screened by cDNA array dot blotting and were sequenced. The putative proteins encoded by the isolated transcripts showed similarities to fungal proteins involved in metabolism, signaling, defense, and virulence, such as the MDR/ABC transporter, glucan 1,3-β-glucosidase, chitin synthase B, copper-sulfate-regulated protein, and serine/threonine phosphatase (calcineurin A). These results provide the first molecular insight into the genes differentially expressed during the adaptation of T. rubrum to a lipidic carbon source.

scholarly journals Distinct Roles of Hemocytes at Different Stages of Infection by Dengue and Zika Viruses in Aedes aegypti Mosquitoes

2021 ◽  
Vol 12 ◽  
Author(s):  
Thiago H. J. F. Leite ◽  
Álvaro G. A. Ferreira ◽  
Jean-Luc Imler ◽  
João T. Marques
Keyword(s):  
Aedes Aegypti ◽  
Cellular Responses ◽  
Innate Immune ◽  
Mosquito Vector ◽  
Phagocytic Function ◽  
Systemic Dissemination ◽  
Latex Beads ◽  
Medical Importance ◽  
Immune Pathways

Aedes aegypti mosquitoes are vectors for arboviruses of medical importance such as dengue (DENV) and Zika (ZIKV) viruses. Different innate immune pathways contribute to the control of arboviruses in the mosquito vector including RNA interference, Toll and Jak-STAT pathways. However, the role of cellular responses mediated by circulating macrophage-like cells known as hemocytes remains unclear. Here we show that hemocytes are recruited to the midgut of Ae. aegypti mosquitoes in response to DENV or ZIKV. Blockade of the phagocytic function of hemocytes using latex beads induced increased accumulation of hemocytes in the midgut and a reduction in virus infection levels in this organ. In contrast, inhibition of phagocytosis by hemocytes led to increased systemic dissemination and replication of DENV and ZIKV. Hence, our work reveals a dual role for hemocytes in Ae. aegypti mosquitoes, whereby phagocytosis is not required to control viral infection in the midgut but is essential to restrict systemic dissemination. Further understanding of the mechanism behind this duality could help the design of vector-based strategies to prevent transmission of arboviruses.

scholarly journals Diverse continuum of CD4+ T-cell states is determined by hierarchical additive integration of cytokine signals

2017 ◽  
Vol 114 (31) ◽  
pp. E6447-E6456 ◽  
Author(s):  
Inbal Eizenberg-Magar ◽  
Jacob Rimer ◽  
Irina Zaretsky ◽  
David Lara-Astiaso ◽  
Shlomit Reich-Zeliger ◽  
...  
Keyword(s):  
Cell Population ◽  
Cellular Responses ◽  
Cd4 T Cell ◽  
Complex Signal ◽  
Signal Integration ◽  
Additional Insight ◽  
Cell Fates ◽  
Linear Approach ◽  
Hierarchical Groups ◽  
Insight Into

During cell differentiation, progenitor cells integrate signals from their environment that guide their development into specialized phenotypes. The ways by which cells respond to complex signal combinations remain difficult to analyze and model. To gain additional insight into signal integration, we systematically mapped the response of CD4+ T cells to a large number of input cytokine combinations that drive their differentiation. We find that, in response to varied input combinations, cells differentiate into a continuum of cell fates as opposed to a limited number of discrete phenotypes. Input cytokines hierarchically influence the cell population, with TGFβ being most dominant followed by IL-6 and IL-4. Mathematical modeling explains these results using additive signal integration within hierarchical groups of input cytokine combinations and correctly predicts cell population response to new input conditions. These findings suggest that complex cellular responses can be effectively described using a segmented linear approach, providing a framework for prediction of cellular responses to new cytokine combinations and doses, with implications to fine-tuned immunotherapies.

Characterization of colorectal-cancer-related cDNA clones obtained by subtractive hybridization screening

1997 ◽  
Vol 123 (8) ◽  
pp. 447-451 ◽  
Author(s):  
Jiang Cao ◽  
Xinhan Cai ◽  
Lei Zheng ◽  
Liyi Geng ◽  
Zhengzheng Shi ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Subtractive Hybridization ◽  
Cdna Clones

scholarly journals Monitoring gene expression changes in bovine oviduct epithelial cells during the oestrous cycle

2004 ◽  
Vol 32 (2) ◽  
pp. 449-466 ◽  
Author(s):  
S Bauersachs ◽  
S Rehfeld ◽  
SE Ulbrich ◽  
S Mallok ◽  
K Prelle ◽  
...  
Keyword(s):  
Gene Expression ◽  
Epithelial Cells ◽  
Expression Profiles ◽  
Cdna Array ◽  
Oestrous Cycle ◽  
Cdna Clones ◽  
Rt Pcr ◽  
Expression Levels ◽  
Starting Point ◽  
Oviduct Epithelial Cells

The oviduct epithelium undergoes marked morphological and functional changes during the oestrous cycle. To study these changes at the level of the transcriptome we did a systematic gene expression analysis of bovine oviduct epithelial cells at oestrus and dioestrus using a combination of subtracted cDNA libraries and cDNA array hybridisation. A total of 3072 cDNA clones of two subtracted libraries were analysed by array hybridisation with cDNA probes derived from six cyclic heifers, three of them slaughtered at oestrus and three at dioestrus. Sequencing of cDNAs showing significant differences in their expression levels revealed 77 different cDNAs. Thirty-seven were expressed at a higher level at oestrus, for the other 40 genes expression levels were higher at dioestrus. The identified genes represented a variety of functional classes. During oestrus especially genes involved in the regulation of protein secretion and protein modification, and mRNAs of secreted proteins, were up-regulated, whereas during dioestrus particularly transcripts of genes involved in transcription regulation showed a slight up-regulation. The concentrations of seven selected transcripts were quantified by real-time RT-PCR to validate the cDNA array hybridisation data. For all seven transcripts, RT-PCR results were in excellent correlation (r>0.92) with the results obtained by array hybridisation. Our study is the first to analyse changes in gene expression profiles of bovine oviduct epithelial cells during different stages of the oestrous cycle, providing a starting point for the clarification of the key transcriptome changes in these cells.

scholarly journals Microarray screening of suppression subtractive hybridization-PCR cDNA libraries identifies novel RNAs regulated by dehydration in the rat supraoptic nucleus

2006 ◽  
Vol 24 (2) ◽  
pp. 163-172 ◽  
Author(s):  
Mohamed T. Ghorbel ◽  
Greig Sharman ◽  
Charles Hindmarch ◽  
Kevin G. Becker ◽  
Tanya Barrett ◽  
...  
Keyword(s):  
General Circulation ◽  
Supraoptic Nucleus ◽  
Subtractive Hybridization ◽  
Biologically Active ◽  
Differentially Expressed ◽  
Cdna Libraries ◽  
Suppressive Subtractive Hybridization ◽  
Cdna Clones ◽  
Microarray Screening ◽  
Mechanistic Basis

The magnocellular neurons (MCNs) of the supraoptic nucleus (SON) and paraventricular nucleus (PVN) of the hypothalamus are the principal site of biosynthesis of prepropeptide precursor of the antidiuretic hormone vasopressin (VP). This precursor is processed during anterograde axonal transportation to terminals in the posterior pituitary gland, where biologically active VP is stored until release into the general circulation in response to physiological activation of the SON by osmotic cues. By binding to V2-type receptors located in the kidney, VP decreases the amount of water lost in urine. Osmotic activation of the SON is accompanied by a dramatic morphological and functional remodeling. We have sought to understand the mechanistic basis of this plasticity in terms of the differential expression of genes. To identify such genes, we adopted an unbiased global approach based on suppressive subtractive hybridization-polymerase chain reaction (SSH-PCR) Using this method, we generated libraries of clones putatively differentially expressed in control vs. dehydrated SON. To rapidly screen these libraries, 1,152 clones were subjected to microarray analysis, resulting in the identification of 459 differentially expressed transcripts. cDNA clones corresponding to 56 of these RNAs were sequenced, revealing many of them to be novel expressed sequence tags (ESTs). Four transcripts were shown by in situ hybridization (ISH) to be significantly up- or downregulated in the SON after dehydration. These genes may represent novel effectors or mediators of SON physiological remodeling.

scholarly journals Rho Family of Ras-Like GTPases in Early-Branching Animals

Cells ◽  
2020 ◽  
Vol 9 (10) ◽  
pp. 2279
Author(s):  
Silvestar Beljan ◽  
Maja Herak Bosnar ◽  
Helena Ćetković
Keyword(s):  
Current Knowledge ◽  
Molecular Switches ◽  
Cellular Responses ◽  
Small Gtpases ◽  
Rho Family Gtpases ◽  
History Of ◽  
Rho Family ◽  
Major Branch ◽  
External Signals ◽  
Insight Into

Non-bilaterian animals consist of four phyla; Porifera, Cnidaria, Ctenophora, and Placozoa. These early-diverging animals are crucial for understanding the evolution of the entire animal lineage. The Rho family of proteins make up a major branch of the Ras superfamily of small GTPases, which function as key molecular switches that play important roles in converting and amplifying external signals into cellular responses. This review represents a compilation of the current knowledge on Rho-family GTPases in non-bilaterian animals, the available experimental data about their biochemical characteristics and functions, as well as original bioinformatics analysis, in order to gain a general insight into the evolutionary history of Rho-family GTPases in simple animals.

Characterization of colorectal-cancer-related cDNA clones obtained by subtractive hybridization screening

1997 ◽  
Vol 123 (8) ◽  
pp. 447-451 ◽  
Author(s):  
Jiang Cao ◽  
Xinhan Cai ◽  
Lei Zheng ◽  
Liyi Geng ◽  
Zhengzheng Shi ◽  
...  
Keyword(s):  
Colorectal Cancer ◽  
Subtractive Hybridization ◽  
Cdna Clones

Role of the Saccharomyces cerevisiae Rad9 protein in sensing and responding to DNA damage

2003 ◽  
Vol 31 (1) ◽  
pp. 242-246 ◽  
Author(s):  
G.W.-L. Toh ◽  
N.F. Lowndes
Keyword(s):  
Saccharomyces Cerevisiae ◽  
Dna Damage ◽  
Cellular Responses ◽  
Eukaryotic Cells ◽  
Genome Damage ◽  
Dna Damage Checkpoints ◽  
Repair Mechanisms ◽  
Sense And Respond ◽  
Insight Into

Eukaryotic cells have evolved surveillance mechanisms, known as DNA-damage checkpoints, that sense and respond to genome damage. DNA-damage checkpoint pathways ensure co-ordinated cellular responses to DNA damage, including cell cycle delays and activation of repair mechanisms. RAD9, from Saccharomyces cerevisiae, was the first damage checkpoint gene to be identified, although its biochemical function remained unknown until recently. This review examines briefly work that provides significant insight into how Rad9 activates the checkpoint signalling kinase Rad53.

scholarly journals Microfluidic measurement of cell motility in response to applied non-homogeneous DC electric fields

2016 ◽  
Vol 5 (2) ◽  
pp. 237-243
Author(s):  
Marisa Rio ◽  
Sharanya Bola ◽  
Richard H. W. Funk ◽  
Gerald Gerlach
Keyword(s):  
Electric Fields ◽  
Soft Lithography ◽  
Cellular Responses ◽  
Biological Processes ◽  
Biological Phenomena ◽  
Inverted Microscope ◽  
Directional Cell Migration ◽  
Insight Into ◽  
Dc Electric Fields

Abstract. Endogenous electric fields (EFs) play an important role in many biological processes. In order to gain an insight into these biological phenomena, externally applied electric fields are used to study cellular responses. In this work, we report the construction and fabrication of a direct current (DC)-electrically stimulated microfluidic biochip and its validation with murine photoreceptor-derived 661 W cells. The presented device has the particularity of offering a non-homogeneous EF environment that best resembles the endogenous electric fields in vitro. The fabrication process is relatively easy, namely by photolithography and soft lithography techniques and, furthermore, it enables live-cell imaging under an inverted microscope. First experimental results reveal cathodal directional cell migration upon applied DC EFs. In summary, the microfluidic biochip has proven biocompatibility and suitability for cellular electrotaxis experiments in non-homogeneous DC electric fields.

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