The role of the human papillomavirus (HPV) in cervical cancer : a review about HPV-induced carcinogenesis and its epidemiology, diagnosis, management and prevention

The human papillomavirus (HPV) was the first virus known to induce carcinogenesis and is associated with cancers of the uterine cervix, anogenital tumors and malignancies of the head and neck. This paper reviews the structure and basic genomic characteristics of the virus and outlines the clinical involvement of the main HPV serotypes, focusing on the carcinogenic role of HPV-16 and 18. The mecha¬nisms that occur in the development of cervical neoplasia due to the oncogenic proteins E6 and E7 which interfere with the regulation of the cell cycle through their interaction with p53 and retinoblastoma protein are described. Epidemiological factors, diagnostic tools and the management of the disease are also reviewed, along with the available vaccines to prevent the viral infection. Insights on current research on involvement of oxidative stress and micro-RNAs in cervical carcinogenesis are also explored as they may unlock new means of diagnosis and treatment in the future.peer-reviewe

Portuguese history storyboard

This paper intends to present relevant facts about the Portuguese culture and history, so as to enable a better understanding of who the Portuguese are and provide an overall perspective of the course of history in this westernmost part of Europe. Although the choice of historical facts was subjective by nature, it is believed it achieves the aim of presenting information in a critical but blithesome way, with a view to also deconstructing national stereotypes, such as that Portuguese people are always late or are crazy about football. Finally, it focuses on some information about the Portuguese language mainly to serve as a term of comparison with other European languages

The Morphological Changes of the IIGP1 Accumulations at the PV Are Accompanied by Loss of <i>T. gondii</i> GRA7 from the PV and its Dissemination throughout the Cytoplasm

<p>(A and B) IFN-γ-induced astrocytes were infected with <i>T. gondii</i> for 2 h (A) or 6 h (B) and stained for IIGP1 (green) and GRA7 (red) (filled arrowheads: IIGP1-negative PVs, open arrowheads: IIGP1-positive PVs). Nuclei were stained with DAPI. (C) Uninduced (left) and IFN-γ-induced (right) astrocytes were infected with <i>T. gondii</i> and stained for GRA7 at 4 h post-infection. Exposure conditions for the two images were the same.</p

The Vacuolar Accumulations of IIGP1 Do Not Reflect Host Cell ER Recruitment by the Parasite

<div><p>Astrocytes were induced with IFN-γ or left untreated and infected with <i>T. gondii</i> 24 h later for 2 h. Cells were fixed and stained for the indicated proteins. (A) Shows a cell that was stained for IIGP1 (red) and calnexin (green). The vacuolar calnexin signal is markedly less concentrated at the PV than IIGP1. (A') shows the same cell as in (A) but with an electronically enhanced IIGP1 signal to reveal its non-vacuolar ER localization. Note the dramatic difference in the ratio of the ER versus PV signal between IIGP1 and calnexin.</p><p>(B and C) Shows astrocytes stained for IIGP1- and the ER-localized PDI. No PDI accumulation at the PV was detected.</p><p>(D) Astrocytes were treated as above but stained for IIGP1 (red) and ERP60 (green). Nuclei were stained with DAPI.</p></div

IFN-γ-Mediated Growth Inhibition and Intracellular Killing of <i>T. gondii</i> Are Accompanied by Accumulation of p47 GTPases at the PV

<div><p>(A) Astrocytes were induced with the indicated concentrations of IFN-γ and infected with <i>T. gondii</i> 24 h later for 68 h. The growth of intracellular parasites was monitored by uracil incorporation assay. (Inset) Lysates of astrocytes induced with the indicated concentrations of IFN-γ for 24 h were probed for IIGP1 protein by Western blotting.</p><p>(B) Untreated or IFN-γ induced astrocytes were infected with <i>T. gondii.</i> After 2 h, extracellular parasites were washed away and cells were either fixed or incubated further for a total of 8 h or 24 h. Shown are the mean values of three independent counts representing a total number of 650–997 cells per time point.</p></div

IIGP1 Associates Directly with the PVM; the Morphology of the Vacuolar IIGP1 Accumulation Changes in a Time-Dependent Manner

<div><p>(A) IFN-γ-induced astrocytes were infected with <i>T. gondii</i> for 6 h, fixed, and subjected to ultra-thin cryosectioning. Sections were labeled for IIGP1 using the 165 antiserum and 10 nm gold particles coupled to protein A. The right side is an enlarged view of the boxed region showing that the IIGP1 label was found in close proximity to the PVM (black arrowhead: PVM; white arrowhead: <i>T. gondii</i> plasma membrane; open arrowhead: <i>T. gondii</i> inner membrane complex; bars 200 nm and 100 nm [inset]).</p><p>(B) IFN-γ-induced astrocytes were fixed at the indicated times post-infection (MOI of 10) and 110–160 IIGP1-positive vacuoles were counted per time point. Shown is the percentage of smooth (white), rough (hatched), and disrupted vacuoles (black).</p><p>(C) IFN-γ-induced astrocytes were infected with <i>T. gondii,</i> fixed 2 h later, and stained for IIGP1 with the 10D7 monoclonal antibody (left) or the 165 antiserum (right). The images show maximum projections of 3D deconvoluted Z-series.</p></div

The Accumulation of p47 GTPases at the PV Is Dependent on Active Invasion by <i>T. gondii</i>

<div><p>(A–D) IFN-γ-induced astrocytes were infected with <i>T. gondii</i> for 2 h, fixed, and stained for IGTP (A), GTPI (B), TGTP1 (C), or IRG-47 (D).</p><p>(E) IFN-γ-induced astrocytes were infected with <i>T. gondii</i> for 2 h, fixed, and stained for IIGP1 (red) and <i>T. gondii</i> (green).</p><p>(F) IFN-γ-induced cells were infected with <i>T. gondii,</i> fixed 24 h later, and stained for IIGP1. White arrowheads point to PVs containing replicating parasites.</p><p>(G and H) IFN-γ-induced cells were infected with heat-killed (G) or live (H) parasites, fixed 2 h later, and stained for IIGP1 (green) and LAMP1 (red). White arrowheads in (G) point to parasites residing in a LAMP1-positive but IIGP1-negative compartment. (H) Shows single sections of a 3D deconvoluted Z-series. Nuclei of host cells and parasites were stained with DAPI.</p></div

Induction of Autophagosomes in Vicinity of Disrupted Vacuoles

<p>Astrocytes were transfected with pEGFP-C3-LC3 and induced with IFN-γ<i>.</i> Cells were infected with <i>T. gondi</i> 24 h later and fixed after 2 h (A) or 6 h (B, C, and D). In cells containing only smooth IIGP1 vacuoles GFP-LC3 remained diffusely distributed throughout the cytoplasm (A). In cells containing disrupted IIGP1 PVs GFP-LC3 localizes to vesicular and filamentous structures that are in close proximity to, but do not engulf the IIGP1-positive PVs (B and C). The arrowheads point to IIGP1-positive PVs. The images shown in (A) were processed by 2D deconvolution. (D) Shows maximum projections of 3D deconvoluted Z-series.</p

IIGP1-Positive Vesicular Structures Are Located at Sites Where the PVM Is Disrupted

<p>Astrocytes were induced with IFN-γ<i>,</i> infected with <i>T. gondii</i> and fixed 6 h later. (A–C) Ultra- thin cryosections were labeled for IIGP1 using the 165 antiserum and 10 nm gold coupled to protein A. The insets in (A) show enlarged views of the boxed regions. The black arrows in the bottom left inset point to IIGP1-labeled vesicular profiles with an apparent electron-dense coat. (D and E) Astrocytes were labeled with the anti-GRA7 mAb and 10 nm gold coupled to protein A. (open white arrowhead: <i>T. gondii</i> inner membrane complex [IMC]; filled white arrowhead: <i>T. gondii</i> plasma membrane; black arrowhead: PVM). Bars: 250 nm.</p

Sensitive Detection of MMP9 Enzymatic Activities in Single Cell-Encapsulated Microdroplets as an Assay of Cancer Cell Invasiveness

Matrix metalloproteinases (MMPs) are typically up-regulated in cancer cells, and play a critical role in assisting metastasis by the breakdown of the extracellular matrix. Here we report an effective strategy for cell invasiveness assay by integrating MMP9 functional activity analysis with single cell-encapsulated microdroplets. A flow focusing capillary microfluidic device has been assembled using “off-the-shelf” fluidic components for high-throughput generation of microdroplets. Tumor cells, MMP9 specific peptides, and other cofactors can be loaded into the device and encapsulated into individual droplets as dynamic microreactors for proteolytic cleavage of the substrate. This design allows for rapid and robust detection of MMP9 enzymatic activities by fluorescent signals in a few minutes. It represents the first demonstration of quantifying MMP9 enzymatic activities at the single cell level with a high throughput performance. This new technique promises functional evaluation of cancer cell invasiveness for important diagnostic or prognostic applications