ETS-1 and ETS-2 are upregulated in a transgenic mouse model of pigmented ocular neoplasm

Purpose: Choroidal melanoma is the most common primary malignant ocular tumor in human adults. Relevant mouse models of human uveal melanoma still remain to be developed. We have studied the transgenic mouse strain, Tyrp-1-TAg, to try to gain insight into possible molecular mechanisms common to pigmented ocular neoplasms occurring spontaneously in the eyes of these mice and human choroidal melanoma. The role of two members of the ETS (E26 avian leukemia oncogene) family of transcription factors, ETS-1 and ETS-2, has been investigated in many cancers but has not yet been studied in ocular tumors. Methods: This is the first study describing the production and distribution of ETS-1 and ETS-2 mRNAs and proteins using in situ hybridization and immunohistochemistry in murine ocular tissue sections of normal control eyes and tumoral eyes from mice of the same age. Using semi-quantitative reverse-transcription polymerase chain reaction (RT–PCR) and western blots experiments, we compared changes in ETS-1 and ETS-2 expression, their protein levels, and the regulation of some of their target gene expressions at different stages of the ocular tumoral progression in the transgenic mouse model, Tyrp-1-TAg, with those in normal eyes from control mice of the same age. Results: In normal control adult mouse eyes, ETS-1 was mostly present in the nuclei of all neuroretinal layers whereas ETS-2 was mostly localized in the cytosol of the cell bodies of these layers with a smaller amount present in the nuclei. Both were found in the retinal pigmentary epithelium (RPE). ETS-1 and ETS-2 mRNA and protein levels were much higher in the ocular tissues of Tyrp-1-TAg mice than in control ocular tissues from wild-type mice. This upregulation was correlated with tumor progression. We also demonstrated upregulation of ETS-1 and ETS-2 target expressions in Tyrp-1-TAg mice when comparing with the same target expressions in control mice. Conclusions: Our findings suggest that ETS-1 and ETS-2 are upregulated in ocular tumors derived from the retinal epithelium and may be involved in one or several signaling pathways that activate the expression of a set of genes involved in ocular tumor progression such as those encoding ICAM-1 (intercellular adhesion molecule-1), PAI-1 (Plasminogen activator inhibitor-1), MCP-1 (monocyte chemoattractant protein-1) and p16 (Cyclin dependent kinase inhibitor 2A)

Changes in Fluoroquinolone-Resistant Streptococcus pneumonia after 7-Valent Conjugate Vaccination, Spain

Four new genotypes appeared in 2006 after childhood vaccination was begun

Skeletal Muscle–Specific Deletion of Lipoprotein Lipase Enhances Insulin Signaling in Skeletal Muscle but Causes Insulin Resistance in Liver and Other Tissues

OBJECTIVE—Skeletal muscle–specific LPL knockout mouse (SMLPL−/−) were created to study the systemic impact of reduced lipoprotein lipid delivery in skeletal muscle on insulin sensitivity, body weight, and composition

The Extended Cleavage Specificity of Human Thrombin

Thrombin is one of the most extensively studied of all proteases. Its central role in the coagulation cascade as well as several other areas has been thoroughly documented. Despite this, its consensus cleavage site has never been determined in detail. Here we have determined its extended substrate recognition profile using phage-display technology. The consensus recognition sequence was identified as, P2-Pro, P1-Arg, P1′-Ser/Ala/Gly/Thr, P2′-not acidic and P3′-Arg. Our analysis also identifies an important role for a P3′-arginine in thrombin substrates lacking a P2-proline. In order to study kinetics of this cooperative or additive effect we developed a system for insertion of various pre-selected cleavable sequences in a linker region between two thioredoxin molecules. Using this system we show that mutations of P2-Pro and P3′-Arg lead to an approximate 20-fold and 14-fold reduction, respectively in the rate of cleavage. Mutating both Pro and Arg results in a drop in cleavage of 200–400 times, which highlights the importance of these two positions for maximal substrate cleavage. Interestingly, no natural substrates display the obtained consensus sequence but represent sequences that show only 1–30% of the optimal cleavage rate for thrombin. This clearly indicates that maximal cleavage, excluding the help of exosite interactions, is not always desired, which may instead cause problems with dysregulated coagulation. It is likely exosite cooperativity has a central role in determining the specificity and rate of cleavage of many of these in vivo substrates. Major effects on cleavage efficiency were also observed for residues as far away as 4 amino acids from the cleavage site. Insertion of an aspartic acid in position P4 resulted in a drop in cleavage by a factor of almost 20 times

DE INVLOED VAN TARIEFSWIJZIGINGEN OP DEN VERKEERSOMVANG BIJ HET AMSTERDAMSCHE TRAMBEDRIJF (Inhoud juli 1932 no. 7)

DE INVLOED VAN TARIEFSWIJZIGINGEN OP DEN VERKEERSOMVANG BIJ HET AMSTERDAMSCHE TRAMBEDRIJF (Inhoud juli 1932 no. 7

Mutation in the human homeobox gene NKX5-3 causes an oculo-auricular syndrome.

Several dysmorphic syndromes affect the development of both the eye and the ear, but only a few are restricted to the eye and the external ear. We describe a developmental defect affecting the eye and the external ear in three members of a consanguineous family. This syndrome is characterized by ophthalmic anomalies (microcornea, microphthalmia, anterior-segment dysgenesis, cataract, coloboma of various parts of the eye, abnormalities of the retinal pigment epithelium, and rod-cone dystrophy) and a particular cleft ear lobule. Linkage analysis and mutation screening revealed in the first exon of the NKX5-3 gene a homozygous 26 nucleotide deletion, generating a truncating protein that lacked the complete homeodomain. Morpholino knockdown expression of the zebrafish nkx5-3 induced microphthalmia and disorganization of the developing retina, thus confirming that this gene represents an additional member implicated in axial patterning of the retina

Dangers of Chemsex: an autopsy case report

Background: ''Chemsex'' is an emerging practice all over the world, sometimes fatal for its users. The victim, a 36-year-old martial arts athlete, was found lifeless lying on his back on the ground in the woods, with a rope tied around the neck, passing over the branch of a tree 2 m from the ground, the other end of the ligature being held in the dead body’s right hand. A third party (a male prostitute) was present at the time of death. Material and methods: A forensic autopsy was performed 48 h after death, including a complete histologic examination of most organs and toxicological analyses. Blood, hair, and organs were analysed for drugs and narcotics (LC-MS/MS and GC-MS), and alcohol was determined by GC-FID. Results and discussion: Autopsy findings showed moderate asphyxia syndrome without clear skin ligature marks, but there were deep cervical contusions. Other contusions were found including deep bruises in the dorsal area and superficial hemorrhagic infiltration of the medial surface of both wrists. Focal bruising of the tongue was found. Organs showed no significant pathology. There was a residual thymus, suggesting chronic drug addiction. Histological examination showed frank signs of hemodynamic shock. Foci of contusions found during autopsy were confirmed, showing no inflammatory reaction. Foci of granular cylinders were present in renal tubules. Heart showed focal non-specific endocardial fibrosis. Toxicological findings were the following ones: (1) peripheral blood: THC at 0.1 ng/mL, THC-COOH at 1 ng/mL, with no 11-OH-THC, indicating last use more than 12 h before death; 3-MMC, NPS-type cathinone at 960 ng/mL, a high, potentially lethal concentration; GHB (gamma-hydroxybutyric acid) at 24 mg/L, a medium concentration, but consistent with last use in the hours before death; ethanol at 0.73 g/L, a relatively low concentration, but which potentiates the toxic effects of the associated molecules. (2) Hair: 3-MMC at 1986 pg/mg, consistent with chronic use in the last few months. (3) Viscera: 3-MMC at 700 ng/mg in the brain, suggesting a cerebral distribution of this molecule. The cardio-femoral ratio of 7.6 was consistent with post mortem release from the heart and/or diffusion from the stomach and/or reservoir organs. Conclusion: We present a case of lethal mechanical asphyxia by atypical hanging, which occurred after recent consumption of 3-MMC and GHB, which contributed to the death in a context of chemsex, which is sometimes associated with sadomasochistic practices such as strangulation. As there was a third-party present at the time of death, according to the police investigation, manner of death was undetermined, as homicide possibly by strangulation was not formally ruled out. This autopsy case illustrates the need to carefully confront police investigations with complete forensic investigations, including exhaustive toxicological analyses