BACKGROUND: The aim of this study was to evaluate the effect of 4-aminopyridine (4-AP) combined with anticholiesterase (antiChE) in antagonizing MgSO4-rocuronium-induced neuromuscualr blockade using a rat hemidiaphragm. METHODS: A hemidiaphragm with phrenic nerve was dissected and was mounted in a bath containing oxygenated Krebs solution. The phrenic nerve was stimulated supramaximally and the twitch response (0.1 Hz) was stabilized for at least 30 minutes. After maximal twitch inhibition by IC95 (concentration of 95% twitch inhibition) of rocuronium and MgSO4 20 mg was achieved, antagonistic effects of 1.6, 16 microgram/ml of edrophonium, 0.1, 1.0 microgram/ml of neostigmine, 0.5, 5.0 microgram/ml of pyridostigmine, and 0.8 microgram/ml of 4-AP combined with each of the above mentioned antiChEs were investigated. RESULTS: Whereas antiChE alone at low concentration partially recovered only the twitch response, 4-AP combined with antiChE recovered both the twitch and train-of-four responses significantly. CONCLUSIONS: 4-AP enhances antagonism of a magnesium-rocuronium induced neuromuscular blockade by edrophonium, neostigmine or pyridostigmine in vitro.
4-Aminopyridine* ; Animals ; Baths ; Cholinesterase Inhibitors* ; Edrophonium ; Neostigmine ; Neuromuscular Blockade* ; Oxygen ; Phrenic Nerve ; Pyridostigmine Bromide ; Rats

BACKGROUND: Overexpression of bcl-2 protein is observed both in follicular lymphoma, in which bcl-2 has usually undergone a translocation t (14;18). The experimental findings that transfection of bcl-2 in to murine lymphoid cells confers resistance to nitrogen mustard and camptothecin by inhibiting apoptosis suggests that bcl-2 overexpression may confer clinical drug resistance in lymphomas. In contrast to bcl-2, p53 arrests cells exposed to DNA-damaging agents in G1 to allow DNA repair or if essential repairs are not possible, promotes apoptosis. Experimentally, loss of p53 function produces cellular resistance to alkylating and topoisomerase-II drug classes, suggesting that loss of p53 function in lymphomas may cause drug resistance. These observations led to the hypothesis that bcl-2 and p53 play a role in the development of drug resistance in lymphoma. Although several studies assessed the association between bcl-2 expression and disease-free survival, they reached conflicting conclusions. METHODS: We analyzed tumor tissue from 42 patients with advanced NHL for p53 and bcl-2 expression and correlation with multiple clinical characteristics, response to therapy and overall survival. Among 42 tumors, 8 (19.0%) tumors had bcl-2 expression and 19 (45.2%) had a p53 overexpression. RESULTS: A significant correlation was found between bcl-2 expression and poor performance, advanced stage (stage III and IV) at diagnosis, and bone marrow involvement in a univariate analysis (P<0.05). A multivariate analysis showed that tumors with bcl-2 expression (>50%) were more likely to be poor prognosis than tumors with negative or week expression (<50%) and to have a shorter long-term survival (28.6% vs 75.5%; P<0.05). However, the expression of p53 did not correlate with any clinical characteristics and overall survival was not influenced by p53 protein expression. CONCLUSION: These results suggest that bcl-2 protein expression in patients with malignant lymphoma appears to be predictive of shorter long-term survival and it might be considered as a strong independent prognostic factor.
Apoptosis ; Bone Marrow ; Camptothecin ; Diagnosis ; Disease-Free Survival ; DNA Repair ; Drug Resistance ; Humans ; Lymphocytes ; Lymphoma* ; Lymphoma, Follicular ; Lymphoma, Non-Hodgkin ; Mechlorethamine ; Multivariate Analysis ; Prognosis ; Transfection

BACKGROUND: Kaposi's sarcoma-associated herpesvirus (KSHV) been shown to be associated with human diseases including Kaposi's sarcoma, pleural effusion lymphoma, multicentric Castleman's disease. The IL-6 may both stimulate myeloma growth and prevent apoptosis of malignant plasma cells. Interestingly, viral IL-6(vIL-6), homolog to human interleukin-6(IL-6) in KSHV genome retains biologic activity. Thus, oncogenic role of the KSHV has been proposed as a pathogenesis of the multiple myeloma. We used ISH to determine the frequency of patients with multiple myeloma and plasmacytosis associated with KSHV-infected BM cells in fresh core biopsies and to determine the correlation between KSHV infection and clinical characteristics. METHODS: Bone marrow(BM) biopsy samples from 16 cases of multiple myeloma, 2 cases of monoclonal gammopathy of undetermined significance(MGUS) were obtained from the pathology division of Soon Chun Hyang University Hospital, Seoul, Korea. Biopsy sample of Kaposi's sarcoma for positive control and BM biopsy samples of myelodysplastic syndrome(MDS) and malignant lymphoma for negative control were obtained. Bitinylated probe to KSHV were prepared with the following sequences: 5' to 3' TGCAGCAGCTGTTGGTGTACCACATATCT. and in situ hybridization (ISH) was performed. RESULTS: Among the 18 patients. Two patients were MGUS and among 16 patients with multiple myeloma, 1 in stage IB disease, 1 stage IIB disease, 8 stage IIIA disease, 4 stage IIIB diseases and 2 in variant of multiple myeloma, extramedullary plasmacytoma. Strong positive signal was detected in nuclei and cytoplasm of the malignant cells of biopsy sample from 1 cases of Kaposi's sarcoma by ISH(positive control). Signal was not detected in BM biopsy samples of 7 cases from MDS and malignant lymphoma(negative control). Among 16 patients with multiple myeloma, 15 demonstrated viral positive cells and 2 cases with MGUS also showed viral positive cells by ISH. Signal was detected in nuclei and cytoplasm of stromal cells. Signal was strongly detected in MGUS than multiple myeloma. Positivity of the KSHV was not related with stage of the patients with multiple myeloma. One patients with multiple myeloma was studied at diagnosis and after chemotherapy. After chemotherapy KSHV was not detected. CONCLUSION: In MGUS and multiple myeloma, KSHV infects the stromal cells of BM rather than malignant plasma cells. On the basis of these data, we have supposed KSHV to play a role in transformation from MGUS to multiple myeloma. Particularly, due to the fact that signal of ISH was strongly detected in MGUS and was not detected in one case with multiple myeloma, it was presumed that KSHV was not major role in already advanced multiple myeloma but statistic significance was not demonstrated because of small numbers of cases. Further studies to reveal the correlation of KSHV and pathogenesis of multiple myeloma are needed.
Apoptosis ; Biopsy ; Cytoplasm ; Diagnosis ; Drug Therapy ; Genome ; Giant Lymph Node Hyperplasia ; Herpesvirus 8, Human ; Humans ; In Situ Hybridization ; Interleukin-6 ; Korea ; Lymphoma ; Multiple Myeloma* ; Paraproteinemias ; Pathology ; Plasma Cells ; Plasmacytoma ; Pleural Effusion ; Sarcoma, Kaposi ; Seoul ; Stromal Cells

PURPOSE: In locally advanced head and neck cancer, radiation therapy is currently unsatisfactory because the end result is often limited regional disease control and survival. A clinical study was carried out to compare the effectiveness between the radiation therapy and the combined chemotherapy and radiation therapy. MATERIALS AND METHOD: Thirty-six patients with previously untreated, locally advanced squamous cell carcinoma of the head and neck were treated with radiotherapy alone and combined chemo-radiotherapy. Induction chemotherapy was administered 2~3 cycles, consisting of intravenous cisplatin (100 mg/m2 on day 1) and 5-fluorouracil (1000 mg/m2/day for 5 days as a continuous infusion) every 4 weeks followed by 7~8 weeks of radiation therapy for a total dose of 60~75 Gy. RESULTS: 1) Among 36 locally advanced head and neck cancer, 17 patients received radiation therapy alone and 19 patients received combined chemo-radiotherapy, respectively. 2) Response rate was 47% (complete response 29%, and partial response 18%) in radiation therapy group and 79% (complete response 37%, and partial response 42%) in combined chemo-radiotherapy group (p<0.05). 3) In median survival, radiation therapy group was 13 months and combined chemo- radiotherapy group was 15 months. Both groups were not significantly different (p>0.05). 4) Treatment related mortality was not noted, but the toxic effects were seen on the half cases of the both groups. Grade II toxicities were similar between the two arms. CONCLUSION: Combined chemotherapy and radiation therapy was more effective in local control but not superior in survival than radiation therapy alone. Continuous evaluation and identification of proper sequence for the therapeutic modality is supposed to prolong the survival of patients.
Arm ; Carcinoma, Squamous Cell ; Cisplatin ; Drug Therapy* ; Fluorouracil ; Head and Neck Neoplasms* ; Head* ; Humans ; Induction Chemotherapy ; Mortality ; Neck ; Radiotherapy

BACKGROUND: Telomeres are repetitive DNA fragments at the termini of chromosomes functioning as stabilizing elements of the DNA. A ribonucleoprotein polymerase, called telomerase, is responsible for the synthesis of such telomeric repeats in embryo and germ cells. During ontogenesis of most normal human somatic cells, there exists a physiological telomerase repressing mechanism. In contrast, malignant cells are characterized by an unlimited progressive potential. Certain physiological agents, such as all-trans retinoic acid (ATRA), 13-cis retinoic acid (13-cisRA), 1alpha-25 dihydroxy vitamin D3 (VD3) and cytosine arabinoside (Ara-C), promote further differentiation of leukemic cells into mature granulocytes and monocytes and subsequently undergo apoptosis. METHODS: To determine if a potential linkage is present between telomerase regulation and the differentiation of malignant hematopoietic cells, the changes in telomerase activity during the maturation of HL-60 cells induced by ATRA, 13-cisRA, VD3 and Ara-C were investigated. RESULTS: Differentiating agents induce HL-60 cells to differentiate into CD11b+ granulocytes and monocyte/macrophages, respectively. Approximately 98% of HL-60 cells acquired the expression of CD11b+ antigen after ATRA, 13-cisRA or Ara-C treatment for 5 days. After 1 day treatment with differentiating agents, no significant difference in telomerase activity was shown between untreated and treated HL-60 cells. A dramatic inhibition of telomerase activity occurred at 3 days treatment of ATRA compared to untreated HL-60 cells. Longer treatment for 5 days with differentiating agents resulted in further decrease of telomerase activity. However, telomerase activity in HL-60 cells was decreased slightly by the VD3 or Ara-C treatment, even though for 5 days. No evidence of differentiation and slight decrease of telomerase activity were observed in ATRA-treated K-562 cells for 5 days. These decrease of telomerase activity were dependent on the incubation time and dose. CONCLUSION: These data clearly show the role of telomerase activity during the differentiation of HL-60 cells. This in vitro model can be useful for studies of the mechanisms controlling telomerase activity and in the search for physiological telomerase modulators.
Apoptosis ; Cholecalciferol ; Cytarabine ; DNA ; Embryonic Structures ; Germ Cells ; Granulocytes ; HL-60 Cells* ; Humans ; Monocytes ; Ribonucleoproteins ; Telomerase* ; Telomere ; Tretinoin

We report a case of recurrent subacute necrotizing lymphadenitis with pancytopenia in 21-years-old-woman. She was admitted to our hospital 4-years interval with fever and abdominal pain. Laboratory findings of the last admission showed pancytopenia, such as WBC 700/microliter, hemoglobin 6.0mmol/L (9.7g/dL), hematocrit 28.8%, and platelet 54,000/microliter. Abdominal CT showed hepatosplenomegaly, enlarged conglomerated lymph nodes in splenic hilum, lesser sac, celiac root, and paraaortic areas. Bone marrow biopsy showed hypocellular marrow (20%) with increased number of megakaryocyte, myeloid hyperplasia, and hemophagocytic histiocytes suggesting infectious process. We performed exploratory laparotomy, and pathologic finding revealed subacute necrotizing lymphadenitis-Kikuchi disease-. She was recovered on 26th hospital day with conservative treatment.
Abdominal Pain ; Biopsy ; Blood Platelets ; Bone Marrow ; Fever ; Hematocrit ; Histiocytes ; Histiocytic Necrotizing Lymphadenitis ; Hyperplasia ; Laparotomy ; Lymph Nodes ; Lymphadenitis* ; Megakaryocytes ; Pancytopenia* ; Peritoneal Cavity ; Tomography, X-Ray Computed

Citrullinemia is a rare inborn error of metabolism of the urea cycle, and was first reported by McMurray, et al. in 1962. It is inherited as an autosomal recessive trait. The normal synthesis of argininosuccinic acid is blocked in this disease due to a deficiency of argininosuccinic acid synthetase(AS), which has been demonstrated in liver cells and fibroblasts. The clinical symptoms are vomiting, lethargy or irritability, convulsion and mental retardation. The diagnosis is made by the finding of an increased plasma citrulline level. Every effort should be made to reduce the blood ammonia level as rapidly as possible before irreversible brain damage occurs. This report describes a case of citrullinemia that was diagnosed through organic acid analysis and amino acid analysis, and reviews the related literatures.
Ammonia ; Argininosuccinic Acid ; Brain ; Citrulline ; Citrullinemia* ; Diagnosis ; Fibroblasts ; Intellectual Disability ; Lethargy ; Liver ; Metabolism ; Plasma ; Seizures ; Urea ; Vomiting

BACKGROUND: Clinical observations and laboratory studies have supported an immune basis for most acquired aplastic anemias, with the majority of patients responding to immunosuppressive therapy. Fas, a member of the tumor necrosis factor (TNF) receptor superfamily is a critical downregulator of cellular immune responses. Proinflammatory cytokines like interferon gamma (IFN-gamma) and TNF-alpha can induce Fas expression and render hematopoietic progenitor cells susceptible to Fas-induced growth suppression and apoptosis. METHODS: In order to investigate the involvement of apoptosis in the pathogenesis of aplastic anemia (AA), we measured the expression of Fas antigen and caspase-3 on bone marrow (BM) mononuclear cells (MNCs) of AA in the presence or absence of IFN-gamma, TNF-alpha, or macrophage inflammatory protein 1-alpha (MIP-1alpha). RESULTS: We confirmed that AA BM MNCs were more apoptotic and highly expressed Fas antigen than normal donors. Stimulation by IFN-gamma, TNF-alpha, or MIP-1alpha increased Fas antigen and caspase-3 expression in AA BM MNCs than BM MNCs of normal donors. Anti-Fas monoclonal antibody enhanced IFN-gamma, TNF-alpha, or MIP-1alpha mediated caspase-3 expression in BM MNCs of normal donors. Among these three cytokines, IFN-gamma enhanced apoptosis most strongly via Fas-caspase-3 pathway. CONCLUSION: These results suggest that Fas signal pathway may play a role in the pathophysiology of aplastic anemia and negative hematopoietic regulators like IFN-gamma can induce apoptosis of bone marrow progenitors in part by Fas induction.
Anemia, Aplastic* ; Antigens, CD95* ; Apoptosis* ; Bone Marrow Cells* ; Bone Marrow* ; Caspase 3 ; Chemokine CCL3 ; Cytokines ; Hematopoietic Stem Cells ; Humans ; Immunity, Cellular ; Interferons ; Signal Transduction ; Tissue Donors ; Tumor Necrosis Factor-alpha

We report a case of malignant proliferating trichilemmal tumor showing multiple distant metastases. The patient demonstrated a round mass in the right occipital area for 12 months and the lesion grew rapidly to assume 8x6.5x4cm in diameter, with areas of superficial erosion and crusting within the recent 3 months. The entire lesion was removed with a wide surgical excision. It recurred on the neck area 4 months after excision and the lesion was removed with surgical resection again. There was evidence of multiple metastases on CNS and mediastinal lymph nodes after 6 months. The patient was treated with cisplatin and etoposide combination chemotherapy and a partial response was achieved.
Adult ; Antineoplastic Agents, Combined/administration & dosage ; Biopsy, Needle ; Brain Neoplasms/therapy ; Brain Neoplasms/secondary* ; Brain Neoplasms/pathology ; Case Report ; Combined Modality Therapy ; Follow-Up Studies ; Head and Neck Neoplasms/therapy ; Head and Neck Neoplasms/surgery ; Head and Neck Neoplasms/pathology* ; Human ; Immunohistochemistry ; Lymphatic Metastasis ; Male ; Neoplasms, Basal Cell/therapy ; Neoplasms, Basal Cell/secondary* ; Neurosurgical Procedures/methods ; Reoperation ; Scalp* ; Skin Neoplasms/therapy ; Skin Neoplasms/surgery ; Skin Neoplasms/pathology* ; Tomography, X-Ray Computed

Synaptotagmin is a Ca2+ sensing protein, which triggers a fusion of synaptic vesicles in neuronal transmission. Little is known regarding the expression of Ca2+ - dependent synaptotagmin isoforms and their contribution to the release of secretory vesicles in mouse and rat parotid acinar cells. We investigated a type of Ca2+ - dependent synaptotagmin and Ca2+ signaling in both rat and mouse parotid acinar cells using RT-PCR, microfluorometry, and amylase assay. Mouse parotid acinar cells exhibited much more sensitive amylase release in response to muscarinic stimulation than did rat parotid acinar cells. However, transient [Ca2+]i increases and Ca2+ influx in response to muscarinic stimulation in both cells were identical, suggesting that the expression or activity of the Ca2+ sensing proteins is different. Seven Ca2+ - dependent synaptotagmins, from 1 to 7, were expressed in the mouse parotid acinar cells. However, in the rat parotid acinar cells, only synaptotagmins 1, 3, 4 and 7 were expressed. These results indicate that the expression of Ca2+ - dependent synaptotagmins may contribute to the release of secretory vesicles in parotid acinar cells.
Synaptotagmins/*metabolism ; Signal Transduction ; Rats ; Protein Isoforms/metabolism ; Parotid Gland/cytology/*metabolism ; Muscarinic Agonists/pharmacology ; Mice ; Exocytosis/drug effects/physiology ; Carbachol/pharmacology ; Calcium/metabolism/*physiology ; Animals ; Amylases/secretion