![]() ![]() They cooperate with different proteins, including other tetraspanins, receptors or signaling proteins to compose functional complexes at the cell surface, designated tetraspanin-enriched microdomains (TEM). Tetraspanins share a comparable structural design, which consists of four hydrophobic transmembrane domains with cytoplasmic and extracellular loops. Tetraspanins, a wide family composed of 33 transmembrane proteins, are associated with different types of proteins through which they arbitrate important cellular processes such as fusion, adhesion, invasion, tissue differentiation and immunological responses. burnetii infection in humans and animals at the population and individual level. Here, we reviewed the current approaches used for the detection of C. The disease has remained largely under-reported, underdiagnosed and as a masked zoonosis and therefore, needs to be explored through well-planned scientific studies for knowing its true status and likely it impact in humans and animals by employing state-of-the-art diagnostics, identifying its diverse and new host range, as well as risk factors involved in different geo-climatic, behavioural and social settings as well as risk groups. burnetii DNA by PCR in various clinical samples have also been widely used. So far serology has been mostly used for testing for C. Therefore, indirect diagnostic tools have been mainly used for its diagnosis. However, it is still to be included among routinely isolated laboratory pathogen, accounting prolonged incubation period (~7 days) and requirement of specific oxygen concentration (2.5% O2). burnetii, using standard routine laboratory culture techniques was impossible until formulation of axenic-based medium. It is mainly transmitted through airborne route in humans and animals. The agent has been classified as a Group B bioterrorism agent by the Centre for Disease Control and Prevention (CDC), and the disease is included in the World Organisation for Animal Health (OIE) list of notifiable diseases. burnetii has been regarded as an obligate intracellular bacterial pathogen. As a causal agent of the one among the 13 global priority zoonoses, having the infectious dose as low as one bacterium, C. Q fever (coxiellosis), caused by Coxiella burnetii, is an emerging or re-emerging zoonotic disease of public health significance and with worldwide distribution. burnetii intensity per cell and to select those cells with large PV (yellow outlines). (The green channel intensity was increased for better visibility of Tf488.) CellProfiler was used to sort infected cells (cyan outlines) by C. burnetii serum and Alexa Fluor 594 conjugated anti-rabbit secondary antibody (red) and Hoescht (blue). Infected HeLa cells loaded with Tf488 (green) were stained with rabbit anti-C. (B) Representative fluorescence micrograph depicting segmentation and sorting of infected cells with large PV. In step 4, CellProfiler modules MeasureObjectIntensity and MeasureObjectShapeSize were used to quantitate multiple size, shape, and intensity parameters for each of the cell objects. In step 3, cell boundaries on the WCS channel were defined using the IdentifySecondaryObjects module to propagate from the host cell nuclei identified in step 2. burnetii objects to remove signal from bacterial nucleic acids, then the IdentifyPrimaryObjects module was applied to define host cell nuclei. In step 2, the Hoescht channel was masked with C. In step 1, the IdentifyPrimaryObjects module was used to define C. burnetii, and whole cell stain (WCS) channels were imported to CellProfiler as gray-scale Tiffs. | Summary of CellProfiler image segmentation and analysis pipeline. ![]()
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