ABSTRACT
The aim of the present study was to describe the findings of postmortem serum and pericardial fluid (PF) cardiac troponin T (cTnT) in various causes of death with regard to the postmortem interval (PMI) and comorbid cardiovascular disease, using 101 autopsy cases with PMI of 8-141 h divided into 9 groups: cardiovascular disease (CVD), other diseases (OD), poisoning (P), asphyxia (A), drowning (D), hypothermia (H), thoracic trauma (TT), other trauma (OT) and fire fatalities (F). The results suggest that cTnT levels may help to differentiate cardiovascular death from poisoning and non-thoracic trauma, as well as to differentiate cardiovascular and other diseases as cause of death from drowning and hypothermia. However, the effect of PMI, unlike comorbid cardiovascular disease, has to be taken into account.
Subject(s)
Pericardium/metabolism , Postmortem Changes , Troponin T/metabolism , Adult , Cardiovascular Diseases/metabolism , Female , Fires , Forensic Pathology , Humans , Hypothermia/metabolism , Male , Middle Aged , Poisoning/metabolism , Wounds and Injuries/metabolismABSTRACT
An experimental study was made of gun-shot wounds inflicted by shots from a modern stub-barrelled pistol 9mm Glock 19. The study has ascertain characteristics differentiating injuries inflicted by shots from this pistol: specific recoil injury, specific topography, intensity and distribution of begrime, gunpowder and metal particles and relevant lesions. It was found that depth of penetration under the skin depends both on shooting distance and the position of the particle from the inlet.
Subject(s)
Forensic Ballistics , Forensic Pathology , Wounds, Gunshot/pathology , Firearms , Humans , Skin/injuriesABSTRACT
Small DNA tumor viruses like human papillomaviruses, simian virus 40, and adenoviruses modulate the activity of cellular tumor suppressor proteins p53 and/or pRB. These viruses replicate as nuclear multicopy extrachromosomal elements during the S phase of the cell cycle, and it has been suggested that inactivation of p53 and pRb is necessary for directing the cells to the S phase. Mouse polyomavirus (Py), however, modulates only the pRB protein activity without any obvious interference with the action of p53. We show here that Py replication was not suppressed by the p53 protein indeed in all tested different mouse cell lines. In addition, E1- and E2-dependent papillomavirus origin replication was insensitive to the action of p53 in mouse cells. We show that in hamster (Chinese hamster ovary) or human (osteosarcoma 143) cell lines the replication of both Py and papillomavirus origins was efficiently blocked by p53. The block of Py replication in human and hamster cells is not caused by the downregulation of large T-antigen expression. The deletion analysis of the p53 protein shows that the RPA binding, proline-rich regulatory, DNA-binding, and oligomerization domains are necessary for p53 action in both replication systems. These results indicate that in mouse cells the p53 protein could be inactive for the suppression of papovavirus replication.
Subject(s)
Bovine papillomavirus 1/physiology , Gene Expression Regulation, Viral , Polyomavirus/physiology , Tumor Suppressor Protein p53/physiology , Virus Replication , Animals , Bovine papillomavirus 1/genetics , CHO Cells , Cell Line , Cricetinae , Fibroblasts , Humans , Immunoblotting , Mice , Plasmids/genetics , Polyomavirus/genetics , Polyomavirus Infections/virology , Replication Origin/genetics , Transfection , Tumor Cells, Cultured , Tumor Suppressor Protein p53/genetics , Tumor Virus Infections/virologyABSTRACT
We describe here the use of two newly mapped bovine papillomavirus type 1 (BPV-1) E2 protein epitopes as tags. We constructed several vector plasmids for overexpression as well as for moderate expression of single- or double-tagged proteins in either Escherichia coli or eukaryotic cells. The new tags were fused to several proteins, and the activity of the tagged proteins was tested in different assays. The tags were shown not to interfere with the function of these proteins in vivo and in vitro. Interaction of the monoclonal antibodies 3F12 and 1E2 with their respective epitopes was specific and had high affinity in a variety of conditions. We have demonstrated that the 3F12 antibody-epitope interaction tolerates high salt concentrations up to 2 M. This permits immunoprecipitation and immunopurification of the tagged proteins in high-salt buffers and reduction of the nonspecific binding of the contaminating proteins. We also provide a protocol for DNA binding and DNase I footprinting assays using the tagged, resin-bound DNA-binding proteins. The BPV-1 E2-derived tags can be recommended as useful tools for detection and purification of proteins.
Subject(s)
DNA-Binding Proteins/immunology , Epitopes , Proteins/isolation & purification , Viral Proteins/immunology , Animals , Antibodies, Monoclonal/immunology , Cattle , DNA/metabolism , DNA, Recombinant , Proteins/analysisABSTRACT
p53 protein was able to block human and bovine papillomavirus DNA amplificational replication while not interfering with Epstein-Barr virus oriP once-per-cell cycle replication. Oligomerization, intact DNA-binding, replication protein A-binding, and proline-rich domains of the p53 protein were essential for efficient inhibition, while the N-terminal transcriptional activation and C-terminal regulatory domains were dispensable for the suppressor activity of the p53 protein. The inhibition of replication was caused neither by the downregulation of expression of the E1 and E2 proteins nor by cell cycle block or apoptosis. Our data suggest that the intrinsic activity of p53 to suppress amplificational replication of the papillomavirus origin may have an important role in the virus life cycle and in virus-cell interactions.
