ABSTRACT
The neurotoxin 6-hydroxydopamine (6-OHDA) is widely used to induce models of Parkinson's disease (PD). We now know that the model induced by 6-OHDA does not include all PD symptoms, although it does reproduce the main cellular processes involved in PD, such as oxidative stress, neurodegeneration, neuroinflammation, and neuronal death by apoptosis. In this review we analyse the factors affecting the vulnerability of dopaminergic neurons as well as the close relationships between neuroinflammation, neurodegeneration, and apoptosis in the 6-OHDA model. Knowledge of the mechanisms involved in neurodegeneration and cell death in this model is the key to identifying potential therapeutic targets for PD.
Subject(s)
Adrenergic Agents/adverse effects , Oxidopamine/adverse effects , Parkinson Disease/physiopathology , Animals , Cells, Cultured , Disease Models, Animal , Dopaminergic Neurons/drug effects , Humans , Nervous System/drug effects , Oxidative Stress/drug effects , Substantia Nigra/drug effectsABSTRACT
AIMS: To create and provide a strain of the food-grade bacterium Lactococcus lactis able to efficiently secrete a modified form of the E7 protein from the human papilloma virus (HPV) type-16. METHODS AND RESULTS: We cloned the coding sequence of a modified E7 (E7m) from the HPV-16 in a plasmid regulated by the strong expression promoter p59. Secretion of the E7m was made by the signal peptide of the usp45 gene. The E7m was detected by Western blot in the cell-free-medium fraction, showing no degradation or aberrant forms. CONCLUSIONS: We constructed a strain of L. lactis able to secrete efficiently a HPV-16 E7 modified protein with diminished transforming activity. SIGNIFICANCE AND IMPACT OF THE STUDY: Human papilloma virus infection is associated with more than 99% of cervical cancers. Immunotherapy targeting E7 to treat HPV-associated cervical malignancies has been demonstrated to be highly efficient. However, native E7 maintains transforming activity. We present this new strain of a food-grade bacterium able to efficiently secrete a HPV-16 E7-modified protein with diminished transforming activity. This new strain could be used as a live vaccine to deliver E7 at a mucosal level and generate antitumour immune responses against HPV-associated tumours.
Subject(s)
Antineoplastic Agents/metabolism , Human papillomavirus 16/metabolism , Lactococcus lactis/metabolism , Papillomavirus E7 Proteins/metabolism , Blotting, Western , Female , Gene Expression Regulation , Human papillomavirus 16/genetics , Humans , Lactococcus lactis/genetics , Papillomavirus E7 Proteins/genetics , Plasmids/genetics , Promoter Regions, Genetic , Protein Sorting SignalsABSTRACT
BACKGROUND: Circulating tumour cells (CTCs) offer a non-invasive approach to obtain and characterise metastatic tumour cells, but their usefulness has been limited by low CTC yields from conventional isolation methods. METHODS: To improve CTC yields and facilitate their molecular characterisation we compared the Food and Drug Administration-approved CellSearch Epithelial Kit (CEK) to a simplified CTC capture method, CellSearch Profile Kit (CPK), on paired blood samples from patients with metastatic breast (n=75) and lung (n=71) cancer. Molecular markers including Human Epidermal growth factor Receptor 2 (HER2) were evaluated on CTCs by fluorescence in situ hybridisation (FISH) and compared to patients' primary and metastatic cancer. RESULTS: The median cell count from patients with breast cancer using the CPK was 117 vs 4 for CEK (P<0.0001). Lung cancer samples were similar; CPK: 145 cells vs CEK:4 cells (P<0.0001). Recovered CTCs were relatively pure (60-70%) and were evaluable by FISH and immunofluorescence. A total of 10 of 30 (33%) breast cancer patients with HER2-negative primary and metastatic tissue had HER2-amplified CTCs. CONCLUSION: The CPK method provides a high yield of relatively pure CTCs, facilitating their molecular characterisation. Circulating tumour cells obtained using CPK technology demonstrate that significant discordance exists between HER2 amplification of a patient's CTCs and that of the primary and metastatic tumour.
Subject(s)
Breast Neoplasms/genetics , Genes, erbB-2/genetics , Immunomagnetic Separation/methods , Neoplastic Cells, Circulating , Breast Neoplasms/blood , Female , Fluorescent Antibody Technique , Gene Amplification , Humans , In Situ Hybridization, FluorescenceSubject(s)
Humans , Male , Female , Emergency Medicine/organization & administration , Nursing/methods , Nursing Care/organization & administration , Nursing Care/standards , Emergency Nursing/methods , Emergency Nursing/standards , Emergency Nursing/trends , Critical Care , Critical Care/methods , Nurse Clinicians/organization & administration , Emergency Medicine , Emergency Nursing/instrumentation , Emergency Nursing/organization & administrationABSTRACT
OBJECTIVE: The aim of the study was evaluate cardiac troponin I (cTnI) determination in patients with chronic renal failure (CRF) and compare with creatine kinase-MB isoenzyme (CK-MB and CK-MB/CK). METHODS: We performed a retrospective study on patients with CRF with MDRD (modification of diet in renal disease study group) < 60 mL/min admitted with suspected myocardial injury by history, physical examination and electrocardiography. cTnI measurement was assessed at admission with the ACCESS analyzer (Beckman). RESULTS: Acute myocardial injury (AMI) was diagnosed in 10% (47/467) patients with cTnI determination > 0.05 ng/mL, while the diagnostic was angina in 9% (41/467) and in 81% (379/467) we finded other diagnostics. In the AMI group, 49% had chest discomfort, 43% diabetes and the mortality was 45%, while in the angina group were 41%, 32% and 7%, respectively. The sensitivity for cTnI with cut-off value > 0.5 ng/mL was 70% and specificity 92%. The number of false positives was 31% (20 patients). DISCUSSION: cTnI is the preferred biomarker for myocardial damage in patients with CRF. Other cut-off value could enhance the sensitivity for AMI.
