Subconfluent ARPE-19 Cells Display Mesenchymal Cell-State Characteristics and Behave like Fibroblasts, Rather Than Epithelial Cells, in Experimental HCMV Infection Studies.

Human cytomegalovirus (HCMV) has a broad cellular tropism and epithelial cells are important physiological targets during infection. The retinal pigment epithelial cell line ARPE-19 has been used to model HCMV infection in epithelial cells for decades and remains a commonly used cell type for studying viral entry, replication, and the cellular response to infection. We previously found that ARPE-19 cells, despite being derived from an epithelial cell explant, express extremely low levels of canonical epithelial proteins, such as E-cadherin and EpCAM.

Accuracy of sonography for non-invasive detection of ovarian and deep endometriosis using #Enzian classification: prospective multicenter diagnostic accuracy study.

Objective: To compare the preoperative detection of endometriosis using transvaginal sonography (TVS) supplemented by transabdominal sonography (TAS) with surgical assessment of disease, using the #Enzian classification for endometriosis.

Methods: This was a prospective multicenter diagnostic accuracy study of women undergoing TVS/TAS and radical surgery for deep endometriosis (DE) at different tertiary referral centers. The localization and grade of severity of the endometriotic lesions and adhesions were described according to the criteria of the #Enzian classification, both at preoperative ultrasound examination and during surgery.

Engineered receptors for human cytomegalovirus that are orthogonal to normal human biology.

A trimeric glycoprotein complex on the surface of human cytomegalovirus (HCMV) binds to platelet-derived growth factor (PDGF) receptor α (PDGFRα) to mediate host cell recognition and fusion of the viral and cellular membranes. Soluble PDGFRα potently neutralizes HCMV in tissue culture, and its potential use as an antiviral therapeutic has the benefit that any escape mutants will likely be attenuated. However, PDGFRα binds multiple PDGF ligands in the human body as part of developmental programs in embryogenesis and continuing through adulthood.

A tumor-specific endogenous repetitive element is induced by herpesviruses.

Tandem satellite repeats account for 3% of the human genome. One of them, Human Satellite II (HSATII), is highly expressed in several epithelial cancers and cancer cell lines. Here we report an acute induction of HSATII RNA in human cells infected with two herpes viruses.

Role of PDGF receptor-α during human cytomegalovirus entry into fibroblasts.

Human CMV (HCMV) exhibits a broad cell tropism that depends on two virion glycoprotein complexes: a trimeric complex (gH/gL/gO) that facilitates viral infection primarily in fibroblasts and a pentameric complex (gH/gL/pUL128-pUL130-pUL131A) that mediates infection in epithelial and endothelial cells. We performed genome-wide CRISPR screens in which the PDGF receptor-α (PDGFRα) was identified as the most significant cellular gene product essential for infection by HCMV virions containing only trimeric complex (trimer-only virus). Trimer-only virus did not enter PDGFRα knockout fibroblasts.

Cellular responses to human cytomegalovirus infection: Induction of a mesenchymal-to-epithelial transition (MET) phenotype.

Human cytomegalovirus (HCMV) is the prototypical human β-herpes virus. Here we perform a systems analysis of the HCMV host-cell transcriptome, using gene set enrichment analysis (GSEA) as an engine to globally map the host-pathogen interaction across two cell types. Our analysis identified several previously unknown signatures of infection, such as induction of potassium channels and amino acid transporters, derepression of genes marked with histone H3 lysine 27 trimethylation (H3K27me3), and inhibition of genes related to epithelial-to-mesenchymal transition (EMT).

Human cytomegalovirus pUL97 kinase induces global changes in the infected cell phosphoproteome.

Replication of human cytomegalovirus (HCMV) is regulated in part by cellular kinases and the single viral Ser/Thr kinase, pUL97. The virus-coded kinase augments the replication of HCMV by enabling nuclear egress and altering cell cycle progression. These roles are accomplished through direct phosphorylation of nuclear lamins and the retinoblastoma protein, respectively.

Sirtuin 4 is a lipoamidase regulating pyruvate dehydrogenase complex activity.

Sirtuins (SIRTs) are critical enzymes that govern genome regulation, metabolism, and aging. Despite conserved deacetylase domains, mitochondrial SIRT4 and SIRT5 have little to no deacetylase activity, and a robust catalytic activity for SIRT4 has been elusive. Here, we establish SIRT4 as a cellular lipoamidase that regulates the pyruvate dehydrogenase complex (PDH).

Crystal structure of the Bcl-XL-Beclin 1 peptide complex: Beclin 1 is a novel BH3-only protein.

Bcl-2 family proteins are key regulators of apoptosis and have recently been shown to modulate autophagy. The tumor suppressor Beclin 1 has been proposed to coordinate both apoptosis and autophagy through direct interaction with anti-apoptotic family members Bcl-2 and/or Bcl-X(L). However, the molecular basis for this interaction remains enigmatic.

Structural analysis of a rhomboid family intramembrane protease reveals a gating mechanism for substrate entry.

Intramembrane proteolysis regulates diverse biological processes. Cleavage of substrate peptide bonds within the membrane bilayer is catalyzed by integral membrane proteases. Here we report the crystal structure of the transmembrane core domain of GlpG, a rhomboid-family intramembrane serine protease from Escherichia coli.

Site-specific transgenesis by Cre-mediated recombination in Drosophila.

Transposons such as P elements are routinely used to stably transfer exogenous DNA (transgenes) into the Drosophila genome. Transgene insertion events, however, are essentially random and are subject to 'position effects' from nearby endogenous regulatory elements. Here we describe a microinjection-based system that uses Cre-mediated recombination to insert transgenes into precise genomic 'landing sites'.

The role of binding site cluster strength in Bicoid-dependent patterning in Drosophila.

The maternal morphogen Bicoid (Bcd) is distributed in an embryonic gradient that is critical for patterning the anterior-posterior (AP) body plan in Drosophila. Previous work identified several target genes that respond directly to Bcd-dependent activation. Positioning of these targets along the AP axis is thought to be controlled by cis-regulatory modules (CRMs) that contain clusters of Bcd-binding sites of different "strengths.

Groucho-dependent repression by sloppy-paired 1 differentially positions anterior pair-rule stripes in the Drosophila embryo.

The Drosophila body plan is composed of a linear array of cephalic, thoracic, and abdominal segments along the anterior posterior axis. The number and positions of individual segments are established by a transcriptional network comprised of maternal effect, gap, pair-rule, and segment polarity genes. The sloppy-paired (slp) locus contains two genes (slp1 and slp2) that are expressed in overlapping striped patterns in the presumptive thorax and abdomen.

Anterior repression of a Drosophila stripe enhancer requires three position-specific mechanisms.

The striped expression pattern of the pair-rule gene even skipped (eve) is established by five stripe-specific enhancers, each of which responds in a unique way to gradients of positional information in the early Drosophila embryo. The enhancer for eve stripe 2 (eve 2) is directly activated by the morphogens Bicoid (Bcd) and Hunchback (Hb). As these proteins are distributed throughout the anterior half of the embryo, formation of a single stripe requires that enhancer activation is prevented in all nuclei anterior to the stripe 2 position.

Age-related reference ranges for fetal foot length.

Aim: The aim of this study was to establish age-dependent reference ranges for fetal foot length with gestation as well as for the femur/foot length ratio, based on a mathematical growth model.

Method: In a prospective cross-sectional study of 610 pregnancies with sonographically confirmed gestational age (< 12 weeks' gestation), fetal biometry was performed in addition to measurements of fetal foot length between 12 and 42 completed weeks of gestation. Reference ranges for the fetal foot and for the femur/foot length ratio were constructed with an overall coverage of 90% of the measured data, using a previously established method of determining reference bands for growth parameters.

Preoperative estimation of screw fixation strength in vertebral bodies.

Study Design: Bone mineral density and bone cross-sectional area of human cadaveric vertebral bodies were investigated radiologically and histologically, respectively. After ventral instrumentation with ventral derotation spondylodesis screws, axial pullout force was measured and compared with radiologic and histologic data.

Objectives: To elucidate how well ventral derotation spondylodesis screw fixation strength can be estimated before surgery by specified applications of dual-energy x-ray absorptiometry, quantitative computed tomography, T2*-relaxation time in magnetic resonance imaging, and histomorphometry.

[Determination of bone quality before spinal instrumentation--value of different in vivo methods].

Objective: The present investigation should elucidate which assessment technique for bone quality is most appropriate to estimate preoperatively fixation strength of instrumental spine fusions.

Methods: VDS-screw fixation strength in 50 human cadaveric vertebral bodies was approximated by means of pullout force assessment. Bone quality was assessed by Dual Energy X-ray Absorptiometry (DEXA), Quantitative Computed Tomography (QCT), MRT and histomorphometry.

Non invasive evaluation of the inflammatory activity in sarcoidosis with high-resolution computed tomography.

Purpose: The value of high resolution computed tomography (HR-CT) in the recognition of pathologic changes of the lung parenchyma, especially in the diagnosis of sarcoidosis, is well established. The importance of these findings in regard to the inflammatory activity is not sufficiently documented, also because a direct histologic correlation is seldom possible.

Method: In a prospective study twenty-one patients with suspected or known sarcoidosis were evaluated.

[Triphasic spiral CT in the diagnosis of liver diseases: comparison with CT arteriography and CT arterio-portography].

Purpose: Assessment of the diagnostic efficacy of triphasic spiral-CT scanning (TPS-CT) for liver disease evaluation. CT arteriography (CTA) and CT arterio-portography (CTAP) were used as reference, methods which together have the highest sensitivity for detecting tumours and the perfusion conditions of the liver.

Material And Methods: 50 TPS-CT and CTA/CTPA were performed in 49 patients.

[Isolated fracture of the trochlea humeri following direct elbow trauma].

Isolated fractures of the Trochlea humeri are only rarely seen because in case of indirect trauma this portion of the elbow is only partially exposed to transmitted forces and in case of direct injury adjacent elements are also involved. On the lateral films a half-moon shaped fragment in front of the Humerus together with joint effusion are indicative of this type of fracture. Depending on the grade of dislocation of the fragment, which has to be shown radiologically, conservative or operative management are chosen.

[Jejunogastric invagination: plain radiologic, sonographic and computed tomographic findings].

Jejunogastric intussusception is a rare complication after gastric surgery and when its diagnosis is delayed a high mortality rate can be expected. This paper reports on the characteristic findings of this entity at ultrasound and CT examination. Both diagnostic procedures have a high sensitivity.

Preoperative staging of breast cancer by palpation, mammography and high-resolution ultrasound.

In breast cancer preoperative determination of the tumor size is important for planning breast-conserving operations. In 100 patients with breast cancer, the preoperative tumor size was measured using clinical, mammographic and sonographic examinations and correlated with the results of a subsequent histological examination. Using a high-resolution real-time system, 98 tumors were visible.