Parkinson's disease (PD) is a common age-related neurodegenerative disorder characterized mainly by motor dysfunction resulting in bradykinesia, rigidity, tremor, gait impairment, and postural instability. The classic pathogenic feature of PD is preferential loss of dopaminergic neurons in the substantia nigra. Downregulation of rRNA transcription is one of major mechanisms to maintain cellular homeostasis under stress conditions. Nucleolar stress has emerged as a component of the degenerative process caused by impaired rRNA transcription and altered nucleolar integrity. Recent study has indicated that the response to stress conditions and quality control mechanisms are impaired in PD, and that metabolic stress may be a trigger mechanism for PD. This review aims to present evidence for a role of nucleolar stress in PD and has summarized mechanisms by which nucleolar stress may play a role in the progression of PD.
Cell Nucleolus ; physiology ; Humans ; Parkinson Disease ; etiology ; physiopathology ; RNA, Ribosomal ; genetics ; Signal Transduction ; Stress, Physiological ; TOR Serine-Threonine Kinases ; physiology

OBJECTIVETo identify the putative specific localization signal sequence of tumor metastasis suppressor gene-1 (TMSG-1) and to explore the mechanism of subcellular localization of TMSG-1 protein.

METHODSVectors expressing green fluorescence protein (GFP) tagged different TMSG-1 fragments were generated and transfected into human embryo kidney 293 (HEK293) cells. The expression of those fusion proteins was detected by Western blotting and their subcellular localizations were observed by laser confocal microscope.

RESULTSGFP was fused with the native TMSG-1(aa1-380) or different fragments including T1 (aa1-70), T2 (aa1-128), T3 (aa129-380), T4 (aa71-128), T5 (aa71-179) and T6 (aa71-380). Anti-GFP Western blotting showed that these fusion proteins were successfully expressed. Under laser confocal microscope, GFP fused with fragment T4 (aa71-128) localized mainly in the nucleolus; GFP fused with fragment T6 (aa71-380) localized diffusely in the nucleus; while other fusion proteins with TMSG-1 (aa1-380) or fragment T1 (aa1-70), T2 (aa1-128), T3 (aa129-380) and T5 (aa71-179) localized in the cytoplasm. Fragment T4(Δ119-128) was generated from T4 with deletion of 10 amino acid of the C terminal. GFP fused with fragment T4(Δ119-128) remained in the nucleus, but no longer in the nucleolus.

CONCLUSIONSThere is a nucleolar localization signal (aa119-128 RRRRNQDRPS) within TMSG-1. This finding may have laid the foundation for further investigations into subcellular localization and function of TMSG-1.

Amino Acid Sequence ; Blotting, Western ; Cell Nucleolus ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Green Fluorescent Proteins ; metabolism ; HEK293 Cells ; Humans ; Membrane Proteins ; genetics ; metabolism ; Microscopy, Confocal ; Nuclear Localization Signals ; Plasmids ; Recombinant Fusion Proteins ; metabolism ; Sphingosine N-Acyltransferase ; genetics ; metabolism ; Transfection ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVETo investigate the relationship between human embryo implantation rates and the zygote morphology and establish zygote morphologic indices that indicate the embryo implantation potential after in vitro fertilization and embryo transfer (IVF-ET).

METHODSSixty-two patients with IVF-ET were enrolled in this study, who were below 35 years of age with endometrium thickness no greater than 8 mm on the day of HCG injection. Embryo transfer was performed on day 3 after oocyte retrieval, and 30 patients with successful implantation of all the embryos transferred (implantation rate of 100%) were allocated into the implantation group, and the other 32 patients with implantation failure (implantation rate of 0) served as the control group. The zygote morphologic characteristics were analyzed for the pronuclei, nucleolar precursor bodies (NPB), polar body, cytoplasmic halo, color and granulation of the cytoplasm, and the results were compared between the two groups.

RESULTSThe implantation rate was significantly higher for embryos with the two pronuclei in close vicinity, central position of the pronuclei in the cytoplasm and comparable size of the two pronuclei. Embryos developed from zygotes with linear arrangement of 3 to 7 NPB in moderate sizes tented to have a higher implantation rate (P<0.05). The implantation rate could be obviously lowered when the cytoplasm contained excessive cytoplasmic granularity (P<0.05). The other morphologic characteristics of the embryos such as the polar bodies, color of the cytoplasm, cytoplasm halo, or vacuoles in the cytoplasm did not significantly impact on the implantation rate.

CONCLUSIONThe morphology of the two pronuclei reflects the quality of the zygote and may help predict the developmental potential of the embryo chosen for transfer on day 3 in IVF.

Adult ; Cell Nucleolus ; metabolism ; Cytoplasmic Granules ; metabolism ; Embryo Implantation ; Embryo Transfer ; Female ; Fertilization in Vitro ; Humans ; Zygote ; cytology

Nucleophosmin (NPM) is a protein that shuttles between the nucleus, nucleoplasm and cytoplasm. NPM gene mutations and aberrant cytoplasmic NPM localization have been recently described in acute myelogenous leukemia (AML) with normal karyotype and in a few myelodysplastic syndromes. Expression of NPM mutant reduces the ability of Arf to initiate a p53 response and to induce cell cycle arrest. Clinical research has revealed that NPM mutations are relative to prognosis and can be used to monitor and quantify minimal residual disease (MRD) in AML patients with normal karyotype, therefore, these findings indicate that nucleophosmin mutations might contribute to illustration of myeloid leukemogenesis. In this paper, the research progress of nucleophosmin mutations in haematological malignancies was reviewed.
Cell Nucleolus ; metabolism ; Hematologic Neoplasms ; genetics ; pathology ; Humans ; Mutation ; Nuclear Proteins ; genetics ; fms-Like Tyrosine Kinase 3 ; genetics

OBJECTIVE: To investigate the effect of oxidative stress on the accumulation of heat shock protein 70 (HSP70) within C2C12 myogenic cells. METHODS: Heat shock response (42 degrees C for 1 h and recovery for 12 h at 37 degrees C) was used to induce the expression of heat shock protein 70. We constructed a recombinant plasmid of HSP70 with enhanced green fluorescent protein (EGFP). After being transfected transiently into C2C12 cells, immunoblotting was used to detect the expression of HSP70 induced by heat shock response and transfection. Immunocytochemistry, fluorescent microscopy and immunoblotting were used to detect the translocation of HSP70. RESULTS: Immunoblotting showed that the overexpression of HSP70 was induced by heat shock response and transient transfenction. HSP70 localized within the cytoplasm of the normal cells, but HSP70 translocated from the cytoplasm to the nucleus and the nucleolus at 1 h after the treatment of oxidative stress (0.5 mmol/L H2O2) by using immunocytochemistry, fluorescent microscopy and immunoblotting for cellular partial proteins. CONCLUSION Oxidative stress may induce the accumulation of heat shock protein 70 within the nucleolus.
Cell Nucleolus ; metabolism ; Cells, Cultured ; HSP70 Heat-Shock Proteins ; metabolism ; Humans ; Myoblasts ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; metabolism ; Oxidative Stress ; physiology

OBJECTIVETo evaluate the value of the invasive front grading (IFG), the proliferative activity of cells at the invasive tumor front (ITF) of squamous cell carcinoma (SCC) of tongue.

METHODSAll the tumors were graded using IFG grading. The expression of PCNA, Ki67 and staining and measurement of AgNOR number were evaluated at the ITF and the other parts of the same tumor.

RESULTSThe expression of PCNA (P < 0.001), Ki67 (P < 0.001), AgNOR number (P < 0.001) at the ITF were higher than those of other parts. The IFG score was significantly correlated with the expression of PCNA (P < 0.05), AgNOR number (P < 0.01) at the ITF. The complete IFG score (P < 0.01) was significant in both univariate and multivariate cox analysis; the AgNOR number at the ITF (P < 0.05) was only significant in univariate analysis (P < 0.05).

CONCLUSIONSThe ITF of SCCs of tongue differentiate poorer than the other parts and is active in proliferation. The complete IFG score and the AgNOR number at the ITF could be a valuable indication to predict the prognosis of SCCs of tongue.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Squamous Cell ; pathology ; Cell Division ; Female ; Humans ; Ki-67 Antigen ; analysis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Nucleolus Organizer Region ; pathology ; Proliferating Cell Nuclear Antigen ; analysis ; Tongue Neoplasms ; pathology

Aneuploidy ; Biomarkers, Tumor ; metabolism ; DNA, Neoplasm ; genetics ; Humans ; Laryngeal Neoplasms ; metabolism ; pathology ; Lymphatic Metastasis ; Nucleolus Organizer Region ; metabolism ; Proliferating Cell Nuclear Antigen ; metabolism ; S Phase

Nucleolar organizer regions (NORs) are loops of DNA which occur in the nucleoli of cells which possess ribosomal RNA genes. The numbers and areas of NORs have been thought to be related to cellular activities. We aimed to investigate the direct relationship between the parameters of AgNORs and cellular proliferative activity using immunohistochemical method with the monoclonal antibody Ki-67, which demonstrates proliferating nuclei. The sequential technique for the simultaneous Ki-67 immunostaining and NOR staining was applied to the same slides of a series of non-Hodgkin's lymphomas (NHL) of the low, intermediate, and high grade type. The number of AgNOR per cell was counted and mean NOR percentage nuclear area (NPNA) was measured by morphometry in both the Ki-67 positive and Ki-67 negative nuclei. The increased immunoreactivity for Ki-67 was found in the high grade than in the low grade non-Hodgkin's lymphoma. This was reflected in the two areas of the palatine tonsils and lymph nodes, the positive cell counts being higher in the follicle center nuclei than in those in the interfollicular compartment. In general the numbers and NPNA of AgNORs were higher in the Ki-67 positive nuclei than in those lacking the antigen in malignant lymphomas as well as in control. The AgNORs numbers and NPNA in controls were the highest in the Ki-67 positive cells in the follicular area and the lowest in the Ki-67 negative cells in the perifollicular area. In malignant lymphomas the numbers and NPNA of AgNORs tended to increase in proportion to their grade in both the Ki-67 positive and negative cells. The numbers of AgNORs of the high grade and the NPNA of the intermediate and the high grade were significantly higher in Ki-67 positive cells than in Ki-67 negative ones. With this double staining method it was now possible to confirm that numbers and NPNA of AgNORs were directly related to the cellular proliferative activity. In maligant lymphoma, among the several parameters of the AgNORs, NPNA in Ki-67 positive cells is proposed to be the most useful marker in determining the prognosis of the patient.
Cell Count ; DNA ; Genes, rRNA ; Humans ; Lymph Nodes ; Lymphoma* ; Lymphoma, Non-Hodgkin ; Nucleolus Organizer Region* ; Palatine Tonsil ; Prognosis

Paraffin-embedded surgical specimens from 27 human astrocytic tumors(7 astrocytomas, 10 anaplastic astrocytomas and 10 glioblastomas) were analyzed immunohistochemically for the presence of p53 protein and proliferation markers of proliferating cell nuclear antigen(PCNA), Ki-67 and agyrophilic nucleolar organizer regions (AgNORs). Approximately 33% of total cases were p53-protein positive. The p53-protein positive nuclei were revealed in 5 cases(50%) of glioblastomas, 4 cases(40%) of anaplastic astocytomas. None of the astrocytomas including 2 pilocytic tumors was p53-protein positive. There were no differences between histological types and p53-protein expression(p=0.0593), however, the more malignant histological features appear to be reflected by a greater incidence of p53 accumulation. In comparative evaluation of p53-protein expression and proliferation indices of PCNA, Ki-67 and AgNORs statistical analysis revealed significant correlation between p53 protein expression and Ki-67 labeling indices only(p=0.0177). p53-protein positive astrocytic tumors showed higher Ki-67 labeling index(10.8+/-6.9%) compared to p53-protein negative tumors(5.3+/-4.3%). In conclusion the malignant histological feature of astrocytic tumors may be associated with p53-protein expression, and among proliferation indices Ki-67 labeling index is correlated with p53-protein expression.
Astrocytoma ; Glioblastoma ; Humans ; Incidence ; Nucleolus Organizer Region ; Proliferating Cell Nuclear Antigen

BACKGROUND: In acute lymphoblastic leukemia (ALL), the importance of argyrophilic nucleolar organizer regions (AgNOR) is not established. METHODS: NOR silver staining was carried out in 74 ALL patients. We analyzed the AgNOR parameters ; counting parameters including number of AgNOR per cell, percentage of cells with one cluster, and area parameters including mean AgNOR area, total AgNOR area, and its percentage of nuclear area by morphometry. Cyclin A index was evaluated by immunohistochemical stain. We compared the AgNOR parameters with cyclin A index and evaluated the differences of AgNOR parameters in accordance with FAB classification, immunophenotype, a new classification of ALL (ALL with maturation), and response to induction chemotherapy. RESULTS: A positive correlation was found between cyclin A index and AgNOR area parameters and a significant negative correlation was found between mean AgNOR area and number of AgNOR per cell (r=-0.433, P=0.000). AgNOR area parameters revealed the highest value in L3. The number of AgNOR per cell in T cell ALL was higher than non-T cell ALL (P=0.011), and the percentage of cells with one cluster was lower (P=0.002). The number of AgNOR per cell in ALLm was lower (P=0.004) and the percentage of cells with one cluster was higher than in typical ALL (P=0.002). In cases achieved complete remission (CR) after induction chemotherapy, the number of AgNOR per cell was higher (P=0.005) and the percentage of cells with one cluster was much lower than in cases failed to achieve CR (P=0.013). CONCLUSIONS: Our study indicates that the AgNOR area parameters are helpful to predict the proliferating activity of leukemic blasts in ALL. It is suggested that the number of AgNOR per cell and the percentage of cells with one cluster provide a valuable information to estimate the response to chemotherapy in ALL.
Classification ; Cyclin A ; Drug Therapy ; Humans ; Induction Chemotherapy ; Nucleolus Organizer Region* ; Precursor Cell Lymphoblastic Leukemia-Lymphoma* ; Silver Staining