Clinical profile of orofacial infections : an experience from two primary care dental practices

Objectives: Orofacial infections are common reasons for dental consultations worldwide. However, there is scarcity of data on clinico-epidemiological profiles reported from primary care dental practices. To address this issue, a study was done to characterize the clinical pattern, age groups affected and sex predilection of orofacial infections in the primary care dental practice. Study design: Clinical data was evaluated from random electronic files of patients for whom antimicrobials were prescribed at two Dental Practices in UK between January 2009 and December 2010. Results: 200 case records were studied. 104 (52%) cases were females. Mean age was 37.2 (+/-15.1) years. 107 (53.5%) cases belonged to age group 21-40 years. Posterior teeth were involved in 112 (56%) cases. Types of disease were as follows: dentoalveolar abscess 63(31.5%), pulpitis 27(13.5%), apical periodontitis 21(10.5%), pericoronitis 21(10.5%), dry socket 13(6.5%), periodontitis 9(4.5%) infected root stump 5(2.5%), facial swelling 5(2.5%) and infections unspecified 36(18%) cases. Conclusions: Orofacial infections affect both sexes equally. 21-40 years is the commonest age-group affected. Dentoalveolar abscess is the commonest infection followed by unspecified infections and pulpiti

A New Inhibitor of Apoptosis from Vaccinia Virus and Eukaryotes

A new apoptosis inhibitor is described from vaccinia virus, camelpox virus, and eukaryotic cells. The inhibitor is a hydrophobic, multiple transmembrane protein that is resident in the Golgi and is named GAAP (Golgi anti-apoptotic protein). Stable expression of both viral GAAP (v-GAAP) and human GAAP (h-GAAP), which is expressed in all human tissues tested, inhibited apoptosis induced by intrinsic and extrinsic apoptotic stimuli. Conversely, knockout of h-GAAP by siRNA induced cell death by apoptosis. v-GAAP and h-GAAP display overlapping functions as shown by the ability of v-GAAP to complement for the loss of h-GAAP. Lastly, deletion of the v-GAAP gene from vaccinia virus did not affect virus replication in cell culture, but affected virus virulence in a murine infection model. This study identifies a new regulator of cell death that is highly conserved in evolution from plants to insects, amphibians, mammals, and poxviruses

Trophoblast organoids as a model for maternal-fetal interactions during human placentation.

The placenta is the extraembryonic organ that supports the fetus during intrauterine life. Although placental dysfunction results in major disorders of pregnancy with immediate and lifelong consequences for the mother and child, our knowledge of the human placenta is limited owing to a lack of functional experimental models1. After implantation, the trophectoderm of the blastocyst rapidly proliferates and generates the trophoblast, the unique cell type of the placenta. In vivo, proliferative villous cytotrophoblast cells differentiate into two main sub-populations: syncytiotrophoblast, the multinucleated epithelium of the villi responsible for nutrient exchange and hormone production, and extravillous trophoblast cells, which anchor the placenta to the maternal decidua and transform the maternal spiral arteries2. Here we describe the generation of long-term, genetically stable organoid cultures of trophoblast that can differentiate into both syncytiotrophoblast and extravillous trophoblast. We used human leukocyte antigen (HLA) typing to confirm that the organoids were derived from the fetus, and verified their identities against four trophoblast-specific criteria3. The cultures organize into villous-like structures, and we detected the secretion of placental-specific peptides and hormones, including human chorionic gonadotropin (hCG), growth differentiation factor 15 (GDF15) and pregnancy-specific glycoprotein (PSG) by mass spectrometry. The organoids also differentiate into HLA-G+ extravillous trophoblast cells, which vigorously invade in three-dimensional cultures. Analysis of the methylome reveals that the organoids closely resemble normal first trimester placentas. This organoid model will be transformative for studying human placental development and for investigating trophoblast interactions with the local and systemic maternal environment.Centre for Trophoblast Reearch Royal Society Dorothy Hodgkin Fellowship Marie Curie Intra-European Fellowshi

v-GAAP Is an Immunomodulator

<div><p>Groups of four BALB/c mice aged between 6 and 8 wk were mock-infected with PBS (•) or infected with 10⁷ pfu of v-GAAP WT (♦), v-ΔGAAP (), or v-GAAP Rev (▴).</p><p>(A) Mice were weighed daily, and results are the mean percentage weight change of each group ± standard error of the mean (SEM) compared with the weight on the day of infection.</p><p>(B) Animals from (A) were monitored daily for signs of illness, which was scored as described [<a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0030017#ppat-0030017-b027" target="_blank">27</a>]. Data are expressed as the mean ± SEM. <i>p</i>-Values were determined by using Student's <i>t</i>-test and indicate mean weight loss (A) or mean signs of illness. *, <i>p</i>-values of mice infected with v-ΔGAAP that were significantly different from those of mice infected with v-GAAP WT and v-GAAP Rev (B).</p><p>(C) Mice infected intranasally with 10⁷ pfu of v-GAAP WT, v-ΔGAAP, or v-GAAP Rev were sacrificed at various days p.i. as indicated. BAL cells were recovered and counted. Columns represent the mean cell yield per mouse ± SEM. Those marked with an asterisk indicate mean cell numbers recovered with v-ΔGAAP that were significantly different (<i>p</i> < 0.05) from those recovered from v-GAAP WT and v-GAAP Rev (three independent experiments with groups of four to five mice).</p></div

v-GAAP Complements for Loss of h-GAAP

<div><p>(A) U2OS-v-GAAP or U2OS-h-GAAP cells were transfected with siRNA oligonucleotides directed against h-GAAP (siRNAs 1–3) or v-GAAP (siRNA1) for 36 h. v-GAAP and h-GAAP expression was assessed by immunofluorescence using an α-HA mAb, followed by secondary Ab conjugated to FITC. The upper and third rows of panels show the fluorescent images, and the second and bottom rows show merged images of the fluorescent panel (immediately above) and an image of the same cells taken by differential interference contrast microscopy. Images shown are representative for at least three independent experiments. Scale bars, 20 μm.</p><p>(B) Cell morphology of U2OS-h-GAAP or U2OS-v-GAAP cells 56 h after transfection with GAAP siRNAs 1–3 was assessed as for <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0030017#ppat-0030017-g004" target="_blank">Figure 4</a>B.</p></div

Downregulation of h-GAAP Causes Cell Death by Appotosis

<div><p>HeLa cells (A–D) or U2OS-h-GAAP cells (E and F) were transfected with siRNA oligos directed to h-GAAP (siRNAs 1–3) or GFP (GFP-siRNA) for 36 h (A and E), or 56 h (B–D and F).</p><p>(A) mRNA level of h-GAAP in HeLa cells was assessed by semi-quantitative RT-PCR (35 cycles) using h-GAAP- or actin-specific primers, respectively. −RT indicates negative control.</p><p>(B) HeLa cell morphology was recorded using a Zeiss Axiovert 200M Live Imaging System with an AxioCam HRc CCD. Images shown are representative of at least three independent experiments.</p><p>(C) HeLa cells were loaded with the fluorescent dye TMRE and loss of the inner mitochondrial membrane potential was assessed by TMRE fluorescence two-color flow cytometry. The graph shows the percentage of cells with decreased TMRE uptake compared to mock-transfected cells. Data are mean percentages from at least three independent experiments (one of which is shown in <a href="http://www.plospathogens.org/article/info:doi/10.1371/journal.ppat.0030017#ppat-0030017-sg002" target="_blank">Figure S2</a>). *, <i>p</i> < 0.05; ** <i>p</i> < 0.005.</p><p>(D) HeLa cells were treated with GFP-siRNA or h-GAAP siRNA1 or were mock-treated and assessed for cleavage of PARP by immunoblotting.</p><p>(E and F) h-GAAP protein expression in U2OS-h-GAAP cells was assessed by immunofluorescence (E) or immunoblotting (F) using an α-HA mAb. In (E), the upper row of panels show the fluorescent images, and the bottom row shows a merged image of the top panels and an image of the same cells taken by differential interference contrast microscopy. Data are representative of three independent experiments. Scale bars, 20 μm.</p></div