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3.
J Hosp Infect ; 84(1): 66-70, 2013 May.
Article in English | MEDLINE | ID: mdl-23561425

ABSTRACT

BACKGROUND: The neonatal intensive care unit (NICU) is a high-risk setting for transmission of meticillin-resistant Staphylococcus aureus (MRSA). Very few studies have investigated the impact of pre-emptive contact precautions applied to outborn neonates transferred to an NICU on the incidence of healthcare-associated (HA)-MRSA transmission. AIM: To assess the efficacy of pre-emptive contact precautions for outborn neonates implemented in an NICU. METHODS: A before-and-after intervention study was conducted in the NICU of Kobe University Hospital. Pre-emptive contact precautions for outborn neonates were introduced in September 2008. The period before the introduction of pre-emptive contact precautions (January 2007-August 2008) was compared with the period after the introduction of pre-emptive contact precautions (September 2008-December 2010). Data for all admitted neonates, neonates who stayed in the NICU for more than three days, length of NICU stay, incidence of MRSA-positive outborn neonates on admission, hand hygiene compliance and incidence of HA-MRSA transmission were compared between the two periods. FINDINGS: There were no significant differences in the percentage of outborn patients admitted to the NICU, percentage of patients who stayed in the NICU for more than three days, length of NICU stay, and incidence of MRSA-positive outborn patients at NICU admission between the groups enrolled before and after the introduction of pre-emptive contact precautions. However, hand hygiene compliance increased, and the incidence of HA-MRSA transmission reduced significantly from 3.5/1000 to 1.3/1000 patient-days after the introduction of pre-emptive contact precautions (P < 0.0001). CONCLUSION: Pre-emptive contact precautions for outborn neonates were effective in reducing the incidence of HA-MRSA transmission in a Japanese NICU.


Subject(s)
Cross Infection/prevention & control , Cross Infection/transmission , Infection Control/methods , Intensive Care Units, Neonatal/standards , Methicillin-Resistant Staphylococcus aureus/isolation & purification , Staphylococcal Infections/prevention & control , Staphylococcal Infections/transmission , Cross Infection/epidemiology , Hand Hygiene , Hospitals , Humans , Incidence , Infant, Newborn , Japan/epidemiology , Length of Stay , Staphylococcal Infections/epidemiology
4.
J Perinatol ; 32(7): 514-9, 2012 Jul.
Article in English | MEDLINE | ID: mdl-21941232

ABSTRACT

OBJECTIVE: To assess renin, aldosterone, human atrial natriuretic peptide (hANP) and brain natriuretic peptide (BNP) levels in cord blood from monochorionic diamniotic (MD) twins with a birthweight (BW) discordance that do not satisfy the criteria of twin-to-twin transfusion syndrome (TTTS). STUDY DESIGN: Cord blood samples were obtained from 28 MD twins without TTTS. They were divided into two groups on the basis of BW discordance as follows: large (>7.5%) and small (7.5%). Cord blood renin, aldosterone, hANP and BNP levels were measured. RESULT: Renin levels in MD twins with a large BW discordance were significantly higher than those in MD twins with a small BW discordance, with no significant differences in aldosterone, hANP and BNP levels. A significant correlation was found between renin levels and BW discordance. CONCLUSION: Renin is activated in MD twins with a BW discordance of >7.5%, even in non-TTTS.


Subject(s)
Fetal Blood/chemistry , Renin/blood , Twins , Aldosterone/blood , Amnion , Atrial Natriuretic Factor/blood , Birth Weight , Chorion , Female , Fetofetal Transfusion/blood , Humans , Infant, Newborn , Male , Natriuretic Peptide, Brain/blood , Pregnancy , Twins, Monozygotic
5.
Neuroscience ; 201: 67-84, 2012 Jan 10.
Article in English | MEDLINE | ID: mdl-22119643

ABSTRACT

Pyramidal neurons of the neocortex are produced from progenitor cells located in the neocortical ventricular zone (VZ) and subventricular zone (SVZ) during embryogenesis. RP58 is a transcriptional repressor that is strongly expressed in the developing brain and plays an essential role in corticogenesis. The expression of RP58 is strictly regulated in a time-dependent and spatially restricted manner. It is maximally expressed in E15-16 embryonic cerebral cortex, localized specifically to the cortical plate and SVZ of the neocortex, hippocampus, and parts of amygdala during brain development, and found in glutamatergic but not GABAergic neurons. Identification of the promoter activity underlying specific expression patterns provides important clues to their mechanisms of action. Here, we show that the RP58 gene promoter is activated prominently in multipolar migrating cells, the first in vivo analysis of RP58 promoter activity in the brain. The 5.3 kb 5'-flanking genomic DNA of the RP58 coding region demonstrates promoter activity in neurons both in vitro and in vivo. This promoter is highly responsive to the transcription factor neurogenin2 (Ngn2), which is a direct upstream activator of RP58 expression. Using in utero electroporation, we demonstrate that RP58 gene promoter activity is first detected in a subpopulation of pin-like VZ cells, then prominently activated in migrating multipolar cells in the multipolar cell accumulation zone (MAZ) located just above the VZ. In dissociated primary cultured cortical neurons, RP58 promoter activity mimics in vivo expression patterns from a molecular standpoint that RP58 is expressed in a fraction of Sox2-positive progenitor cells, Ngn2-positive neuronal committed cells, and Tuj1-positive young neurons, but not in Dlx2-positive GABAergic neurons. Finally, we show that Cre recombinase expression under the control of the RP58 gene promoter is a feasible tool for conditional gene switching in post-mitotic multipolar migrating young neurons in the developing cerebral cortex.


Subject(s)
5' Flanking Region/genetics , Cerebral Ventricles/cytology , Neurogenesis/genetics , Repressor Proteins/genetics , Stem Cells/physiology , Animals , Basic Helix-Loop-Helix Transcription Factors/deficiency , Cell Differentiation , Cell Movement , Cells, Cultured , Cerebral Cortex/cytology , Chick Embryo , Chlorocebus aethiops , Electroporation/methods , Embryo, Mammalian , Gene Expression Regulation, Developmental/genetics , Homeodomain Proteins/metabolism , Luminescent Proteins/genetics , Mice , Mice, Knockout , Nerve Tissue Proteins/deficiency , Nerve Tissue Proteins/genetics , Nerve Tissue Proteins/metabolism , RNA, Messenger , SOXB1 Transcription Factors/metabolism , Transcription Factors/metabolism , Transfection , Tubulin/metabolism
6.
Br J Cancer ; 102(5): 844-51, 2010 Mar 02.
Article in English | MEDLINE | ID: mdl-20145621

ABSTRACT

BACKGROUND: Gastric cancer cells frequently metastasise, partly because of their highly invasive nature. Transforming growth factor-beta (TGF-beta) receptor signalling is closely associated with the invasion of cancer cells. The aim of this study was to clarify the effect of a TGF-beta receptor (TbetaR) phosphorylation inhibitor on the invasiveness of gastric cancer cells. METHODS: Four gastric cancer cell lines, including two scirrhous-type cell lines and two non-scirrhous-type cell lines, were used. A TbetaR type I (TbetaR-I) kinase inhibitor, Ki26894, inhibits the phosphorylation of Smad2 at an ATP-binding site of TbetaR-I. We investigated the expression levels of TbetaR and phospho-Smad2, and the effects of TGF-beta in the presence or absence of Ki26894 on Smad2 phosphorylation, invasion, migration, epithelial-to-mesenchymal transition (EMT), Ras homologue gene family member A (RhoA), ZO-2, myosin, and E-cadherin expression of gastric cancer cells. RESULTS: TbetaR-I, TbetaR-II, and phospho-Smad2 expressions were found in scirrhous gastric cancer cells, but not in non-scirrhous gastric cancer cells. Ki26894 decreased Smad2 phosphorylation induced by TGF-beta1 in scirrhous gastric cancer cells. Transforming growth factor-beta1 upregulated the invasion, migration, and EMT ability of scirrhous gastric cancer cells. Transforming growth factor-beta1 significantly upregulated the activity of RhoA and myosin phosphorylation, whereas TGF-beta1 decreased ZO-2 and E-cadherin expression in scirrhous gastric cancer cells. Interestingly, Ki26894 inhibited these characteristics in scirrhous gastric cancer cells. In contrast, non-scirrhous gastric cancer cells were not affected by TGF-beta1 or Ki26894 treatment. CONCLUSION: A TbetaR-I kinase inhibitor decreases the invasiveness and EMT of scirrhous gastric cancer cells. Ki26894 is therefore considered to be a promising therapeutic compound for the metastasis of scirrhous gastric carcinoma.


Subject(s)
Activin Receptors, Type I/antagonists & inhibitors , Adenocarcinoma, Scirrhous/drug therapy , Protein Serine-Threonine Kinases/antagonists & inhibitors , Receptors, Transforming Growth Factor beta/antagonists & inhibitors , Stomach Neoplasms/drug therapy , Activin Receptors, Type I/pharmacology , Adenocarcinoma, Scirrhous/pathology , Animals , Blotting, Western , Cell Movement/drug effects , Female , Mice , Mice, Inbred BALB C , Mice, Nude , Neoplasm Invasiveness , Phosphorylation/drug effects , Protein Serine-Threonine Kinases/metabolism , Receptor, Transforming Growth Factor-beta Type I , Receptor, Transforming Growth Factor-beta Type II , Receptors, Transforming Growth Factor beta/metabolism , Signal Transduction/drug effects , Smad2 Protein/metabolism , Smad3 Protein/metabolism , Stomach Neoplasms/pathology , Transforming Growth Factor beta/pharmacology , Wound Healing/drug effects
7.
Br J Cancer ; 101(7): 1100-6, 2009 Oct 06.
Article in English | MEDLINE | ID: mdl-19738610

ABSTRACT

BACKGROUND: Vascular endothelial growth factor receptor-3 (VEGFR-3) signalling mediates lymphangiogenesis and lymphatic invasion; however, the effect of VEGFR-3 inhibition on the lymph node (LN) metastasis remains unclear. The aim of this study is to clarify the benefit of a VEGFR-3 inhibitor Ki23057 for LN metastasis. METHODS: Ki23057 was administered orally to gastric cancer models created by orthotopic inoculation of diffuse-type gastric cancer cells, OCUM-2MLN. The effects of Ki23057 on lymphatic vessel invasion, lymphatic vessel density, and VEGFR-3 phosphorylation were examined by immunostaining or immunoblotting. RESULTS: Ki23057 inhibited the autophosphorylation of VEGFR-3, with IC50 values of 4.3 nM in the cell-free kinase assay. Murine gastric cancer models created by the orthotopic inoculation of OCUM-2MLN cells showed the diffusely infiltrating growth and frequently developed LN metastasis. The oral administration of Ki23057 significantly (P<0.01) reduced the size of orthotopic tumours and the number of the metastatic LN in gastric cancer models. The degree of lymphatic invasion and lymphangiogenesis was significantly (P<0.05) lower in the gastric tumours treated by Ki23057. Ki23057 inhibited the phosphorylation of VEGFR-3 of lymphatic endothelial cells in gastric tumours. CONCLUSION: The inhibition of lymphangiogenesis targeting VEGFR-3 phosphorylation is a therapeutic strategy for inhibiting LN metastasis of diffuse-type gastric cancer.


Subject(s)
Lymphatic Metastasis/prevention & control , Quinolines/therapeutic use , Stomach Neoplasms/drug therapy , Vascular Endothelial Growth Factor Receptor-3/antagonists & inhibitors , Animals , Cell Line, Tumor , Female , Glycoproteins/analysis , Humans , Lymphangiogenesis/drug effects , Membrane Transport Proteins , Mice , Mice, Inbred BALB C , Phosphorylation , Quinolines/pharmacology , Stomach Neoplasms/blood supply , Stomach Neoplasms/pathology , Vascular Endothelial Growth Factor C/analysis , Vascular Endothelial Growth Factor Receptor-3/metabolism
8.
J Hazard Mater ; 167(1-3): 224-9, 2009 Aug 15.
Article in English | MEDLINE | ID: mdl-19185994

ABSTRACT

This paper presents the results concerning the degradation of the pesticide carbaryl comparing two methods: electrochemical (EC) and photo-assisted electrochemical (PAEC). The experimental variables of applied current density, electrolyte flow-rate and initial carbaryl concentration were investigated. The results demonstrate that the electrochemical degradation of carbaryl was greatly enhanced when simultaneous UV light was applied. The greatest difference between the PAEC and EC method was apparent when lower current densities were applied. The extent of COD removal was much enhanced for the combined method, independent of the applied current density. It should be noted that the complete removal of carbaryl was achieved with out the need to add NaCl to the reaction mixture, avoiding the risk of chlorinated organic species formation.


Subject(s)
Carbaryl/chemistry , Electrochemical Techniques/methods , Environmental Restoration and Remediation/methods , Cholinesterase Inhibitors , Decontamination , Electrodes , Environmental Pollutants/chemistry , Insecticides , Photochemical Processes , Ultraviolet Rays
9.
Kyobu Geka ; 61(12): 1035-8, 2008 Nov.
Article in Japanese | MEDLINE | ID: mdl-19048903

ABSTRACT

Myxomas are account for approximately half of primary cardiac tumors, and 75% cases originate in left atrium. We report our experience of a right atrial myxoma. A 68-year-old woman was referred to us due to anorexia, general fatigue and facial edema. Echocardiogram, computed tomography (CT), magnetic resonance imaging (MRI), and catheter angiocardiogram revealed a huge tumor in right atrium. The tumor was resected completely with the attached right atrial free wall under cardiopulmonary bypass. Pathological examination showed myxomatous tissue. Postoperative course was uneventful. She discharged the hospital on the 37th day after the operation, and is now doing well without any symptoms.


Subject(s)
Heart Neoplasms/surgery , Myxoma/surgery , Aged , Cardiopulmonary Bypass , Female , Heart Atria , Humans
10.
Environ Sci Technol ; 41(20): 7120-5, 2007 Oct 15.
Article in English | MEDLINE | ID: mdl-17993157

ABSTRACT

This paper presents the study of the degradation of the pesticide atrazine, employing photoassisted electrochemical methods at a dimensionally stable anode of nominal composition Ti/Ru0.3Ti0.7O2. All experiments were performed in a flow cell, and the effects of current density, electrolyte flow rate, as well the use of different supporting electrolytes are reported. The results indicate that the energy per order (E(EO)/kW h m(-3) order(-1)) is substantially reduced when the photochemical and electrochemical techniques are combined. It is observed that complete atrazine removal is achieved at low current densities when the combined method is employed, thus reducing the energy required to operate the electrochemical system. The study of the electrolyte flow rate through the cell indicates that the photoassisted removal of atrazine is controlled by mass transfer. The degradation of commercial atrazine solutions is also presented, and the results show that the efficiency of atrazine removal is reduced compared with that of simulated solutions, due to the presence of the additional components present in the commercial formula.


Subject(s)
Atrazine/chemistry , Electrochemistry , Electrodes , Photochemistry , Ruthenium/chemistry , Titanium/chemistry , Oxidation-Reduction
11.
Int J Gynecol Cancer ; 16(3): 1309-13, 2006.
Article in English | MEDLINE | ID: mdl-16803522

ABSTRACT

Prognoses in cases of uterine cervical squamous cell carcinoma treated with radiotherapy were investigated in association with immunohistochemical expression of an angiogenic factor, thymidine phosphorylase (TP). Forty-six cases of uterine cervical squamous cell carcinoma mainly treated with radiotherapy during 1992-2001 at our clinic were studied. All were diagnosed as stages IIB to IVA. Paraffin-embedded biopsy specimens excised before radiotherapy were stained immunohistochemically using anti-TP monoclonal antibody. The extent of staining in both tumor and interstitial cells was graded as (-), (+/-), (+), and (2+). Specimens with TP expression levels of (2+) and (+) were regarded as positively stained and those with TP expression levels of (+/-) and (-) as negatively stained. The efficacy of radiotherapy in both groups was analyzed by the Kaplan-Meier method. With tumor cells, 5-year survival rates for the positive (n= 38) and negative (n= 8) staining groups were 73.9% and 42.9%, respectively; the rate being significantly higher for the TP-positive group (log rank, P= 0.0096). Contrarily, with staining for interstitial cells, the 5-year survival rates for the positive (n= 20) and negative (n= 26) staining groups were 74.1% and 64.6%, respectively, with no significant difference (log rank, P= 0.406). The efficacy of radiotherapy in the group with positive staining of tumor cells was significantly better than in the negative staining group. Immunohistochemical expression of TP in tumor cells is suggested as a useful prognostic factor for uterine cervical squamous cell carcinomas treated with radiotherapy. Choosing therapy for individual cases by referring to factors including TP expression should contribute to an improved prognosis.


Subject(s)
Carcinoma, Squamous Cell/enzymology , Thymidine Phosphorylase/metabolism , Uterine Cervical Neoplasms/enzymology , Aged , Aged, 80 and over , Biomarkers, Tumor/analysis , Carcinoma, Squamous Cell/radiotherapy , Chemotherapy, Adjuvant/methods , Cisplatin/therapeutic use , Combined Modality Therapy , Female , Gene Expression , Humans , Immunohistochemistry/methods , Middle Aged , Neoplasm Staging , Prognosis , Survival Analysis , Uterine Cervical Neoplasms/radiotherapy
12.
Oncol Rep ; 15(4): 791-5, 2006 Apr.
Article in English | MEDLINE | ID: mdl-16525660

ABSTRACT

To ascertain the implications of loss of imprinting (LOI) of the insulin-like growth factor II gene (IGF2) for carcinogenesis, the precise frequency of LOI in colorectal carcinoma was examined using a laser capture microdissection method, and compared to the matched normal colorectal mucosa. LOI was examined by PCR-restriction fragment length polymorphism in combination with direct sequencing. The status was assigned as imprinting when PCR-RFLP showed only one band or sequence with a single peak, otherwise cases were classified as LOI. LOI was found in 13 of 24 informative cases of carcinoma (54%), which was higher than the ratios reported previously. LOI was also found in the normal colorectal mucosae in 14 cases (58%). The LOIs in carcinomas and in the normal mucosae were closely correlated: 10 of 13 LOI-positive carcinomas showed LOI in the matched normal mucosae. These results suggest that LOI of IGF2 in colorectal carcinoma and LOI in the background mucosa play important roles in carcinogenesis.


Subject(s)
Colorectal Neoplasms/genetics , Genomic Imprinting/genetics , Insulin-Like Growth Factor II/genetics , Microdissection/methods , Binding Sites/genetics , Colorectal Neoplasms/pathology , DNA, Neoplasm/genetics , DNA, Neoplasm/metabolism , Deoxyribonucleases, Type II Site-Specific/metabolism , Humans , Intestinal Mucosa/metabolism , Intestinal Mucosa/pathology , Lasers , Polymorphism, Genetic , Polymorphism, Restriction Fragment Length
13.
Leukemia ; 19(6): 930-5, 2005 Jun.
Article in English | MEDLINE | ID: mdl-15815726

ABSTRACT

Constitutively active internal tandem duplication (ITD) in the juxtamembrane domain of Fms-like tyrosine kinase 3 (FLT3), a type III receptor tyrosine kinase, is the most common molecular defect associated with acute myeloid leukemia. Its presence confers a poor outcome in patients with acute myeloid leukemia who receive conventional chemotherapy. FLT3-ITD has therefore been considered to be an attractive molecular target for a novel therapeutic modality. We describe here the identification and characterization of Ki23819 as a novel FLT3 inhibitor. Ki23819 suppressed proliferation and induced apoptosis of FLT3-ITD-expressing human leukemia cell lines. The growth-inhibitory effect of Ki23819 on MV4-11 cells was superior to that of SU11248, another FLT3 inhibitor (IC(50)<1 vs 3-10 nM). Ki23819 inhibited the autophosphorylation of FLT3-ITD more efficiently than that of wild-type FLT3. FLT3-ITD-dependent activation of the downstream signaling proteins ERK and STAT5 was also inhibited within similar concentration ranges. Thus, Ki23819 is a potent in vitro inhibitor of FLT3.


Subject(s)
Leukemia, Myeloid/drug therapy , Protein Kinase Inhibitors/pharmacology , Proto-Oncogene Proteins/antagonists & inhibitors , Proto-Oncogene Proteins/genetics , Quinolines/pharmacology , Receptor Protein-Tyrosine Kinases/antagonists & inhibitors , Receptor Protein-Tyrosine Kinases/genetics , Urea/analogs & derivatives , Urea/pharmacology , Acute Disease , Apoptosis/drug effects , Cell Division/drug effects , DNA-Binding Proteins/metabolism , Extracellular Signal-Regulated MAP Kinases/metabolism , HL-60 Cells , Humans , K562 Cells , Leukemia, Myeloid/genetics , Leukemia, Myeloid/metabolism , Milk Proteins/metabolism , Mutation , Phosphorylation/drug effects , Protein Kinase Inhibitors/chemistry , Proto-Oncogene Proteins/metabolism , Quinolines/chemistry , Receptor Protein-Tyrosine Kinases/metabolism , STAT5 Transcription Factor , Trans-Activators/metabolism , Urea/chemistry , fms-Like Tyrosine Kinase 3
14.
Clin Lab Haematol ; 24(1): 33-6, 2002 Feb.
Article in English | MEDLINE | ID: mdl-11843896

ABSTRACT

Flow cytometry (FC) is widely utilized in the diagnosis of lymphoma and the light chain ratio (LCR) is especially useful in the diagnosis of B-cell malignancy. In this study we analysed, retrospectively, the predictive value of the LCR in the diagnosis of B-cell lymphoma in 105 consecutive patients with persistent lymph node enlargement or extranodal masses who underwent biopsy. We used a receiver-operating characteristic curve to establish a LCR threshold value of 2.0. The specificity, sensitivity, positive and negative predictive values were 92.3%, 73.1%, 90% and 77%, respectively. We concluded that determination of LCR is a useful adjunct to pathological diagnosis.


Subject(s)
Immunoglobulin Light Chains/analysis , Lymphoma, B-Cell/diagnosis , Adolescent , Adult , Aged , Aged, 80 and over , Biopsy , Child , Child, Preschool , Female , Flow Cytometry , Humans , Immunoglobulin Light Chains/classification , Immunoglobulin kappa-Chains/analysis , Immunoglobulin lambda-Chains/analysis , Male , Middle Aged , Predictive Value of Tests , ROC Curve , Retrospective Studies , Sensitivity and Specificity
15.
Ann Hematol ; 81(1): 33-6, 2002 Jan.
Article in English | MEDLINE | ID: mdl-11807633

ABSTRACT

The range of survival duration in myeloma patients is wide and several percent of patients live longer than 10 years. Therefore, a precise prediction of survival for the individual patient is required to decide treatment. We evaluated possible prognostic factors at diagnosis for 116 Japanese patients with multiple myeloma. Twelve parameters reported to affect survival were analyzed using a log rank test and stepwise Cox proportional hazards regression. Factors identified as adversely affecting survival were age over 60 years, male sex, blood hemoglobin less than 8.5 g/dl, platelets less than 100 x 10(9)/l, serum creatinine level more than 2.0 mg/dl, serum C-reactive protein (CRP) level more than 6.0 mg/l, and serum beta2-microglobulin level more than 6.0 mg/l. Among them, only high age and high serum CRP level were independently prognostic for poor survival. In conclusion, we have established a simple prognostic model for Japanese myeloma patients only, using factors that can be determined in routine examinations without the need of subjective information.


Subject(s)
Multiple Myeloma , Adult , Aged , Aged, 80 and over , C-Reactive Protein/analysis , Creatinine/blood , Female , Hemoglobins/analysis , Humans , Male , Middle Aged , Multiple Myeloma/blood , Multiple Myeloma/mortality , Multiple Myeloma/physiopathology , Multivariate Analysis , Platelet Count , Prognosis , Retrospective Studies , Survival Analysis , beta 2-Microglobulin/blood
16.
Biochim Biophys Acta ; 1550(1): 70-80, 2001 Nov 26.
Article in English | MEDLINE | ID: mdl-11738089

ABSTRACT

A novel antimicrobial peptide, anoplin, was purified from the venom of the solitary wasp Anoplius samariensis. The sequence was mostly analyzed by mass spectrometry, which was corroborated by solid-phase synthesis. Anoplin, composed of 10 amino acid residues, Gly-Leu-Leu-Lys-Arg-Ile-Lys-Thr-Leu-Leu-NH2, has a high homology to crabrolin and mastoparan-X, the mast cell degranulating peptides from social wasp venoms, and, therefore, can be predicted to adopt an amphipathic alpha-helix secondary structure. In fact, the circular dichroism (CD) spectra of anoplin in the presence of trifluoroethanol or sodium dodecyl sulfate showed a high content, up to 55%, of the alpha-helical conformation. A modeling study of anoplin based on its homology to mastoparan-X supported the CD results. Biological evaluation using the synthetic peptide revealed that this peptide exhibited potent activity in stimulating degranulation from rat peritoneal mast cells and broad-spectrum antimicrobial activity against both Gram-positive and Gram-negative bacteria. Therefore, this is the first antimicrobial component to be found in the solitary wasp venom and it may play a key role in preventing potential infection by microorganisms during prey consumption by their larvae. Moreover, this peptide is the smallest among the linear alpha-helical antimicrobial peptides hitherto found in nature, which is advantageous for chemical manipulation and medical application.


Subject(s)
Anti-Bacterial Agents/isolation & purification , Oligopeptides/chemistry , Oligopeptides/isolation & purification , Wasp Venoms/chemistry , Wasp Venoms/isolation & purification , Amino Acid Sequence , Animals , Anti-Bacterial Agents/chemistry , Anti-Bacterial Agents/pharmacology , Antimicrobial Cationic Peptides , Cell Degranulation , Chromatography, High Pressure Liquid , Circular Dichroism , Female , Mast Cells/drug effects , Mast Cells/physiology , Microbial Sensitivity Tests , Models, Molecular , Oligopeptides/pharmacology , Rats , Sequence Alignment , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Wasp Venoms/pharmacology , Wasps
18.
Photochem Photobiol ; 74(4): 579-86, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11683038

ABSTRACT

Bcl-2 is a member of the large Bcl-2 family and protects cells from apoptosis. Ultraviolet B (UVB) irradiation induces apoptosis of keratinocytes that is known as "sunburn cells." Previously we reported that UVB irradiation induces apoptosis accompanied by sequential activation of caspase 8, 3 and 1 in keratinocytes, and that the process is inhibited by various caspase inhibitors. Using bcl-2-expressing adenovirus vector we investigated the effect of Bcl-2 on UVB-induced apoptosis. Adenovirus vector efficiently introduced bcl-2 gene in cultured normal mouse keratinocytes (NMK cells); almost all NMK cells (1 x 10(6)) were transfected at 1 x 10(8) plaque-forming unit (PFU)/mL. Bcl-2-transfected NMK cells were significantly resistant to UVB-induced apoptosis with the suppressive effect dependent on the Bcl-2 expression level. Following UVB irradiation caspase 8, 3 and 9 activities were stimulated in NMK cells, whereas in bcl-2-transfected cells only caspase 8, but not caspase 3 or 9, activity was stimulated. In order to investigate the effect of Bcl-2 in vivo topical application of Ad-bcl-2 on tape-stripped mouse skin was performed. Following the application Bcl-2 was efficiently overexpressed in almost all viable keratinocytes. The expression was transient with the maximal expression of Bcl-2 on the first day following the application of 1 x 10(9) PFU in 200 microL. The introduced Bcl-2 remained at least for 6 days. UVB irradiation (1250 J/m2) induced apoptosis within 12 h and the maximal effect was observed at 24 h in control mouse skin. Both bcl-2-transfected and topical caspase 3 inhibitor-treated mice skin were resistant to UVB-induced apoptosis. The suppressive effect of Bcl-2 was more potent than that of caspase 3 inhibitor application. Topical application of empty adenovirus vector alone had no effect on Bcl-2 expression or UVB-induced apoptosis. These results indicate that adenovirus vector is an efficient gene delivery system into keratinocytes and that Bcl-2 is a potent inhibitor of UVB-induced apoptosis both in vitro and in vivo.


Subject(s)
Apoptosis/radiation effects , Caspase 1/metabolism , Caspases/metabolism , Cells, Cultured/radiation effects , Enzyme Inhibitors/metabolism , Gene Expression Regulation/radiation effects , Keratinocytes , Proto-Oncogene Proteins c-bcl-2 , Skin , Ultraviolet Rays/adverse effects , Adenoviridae , Animals , Blotting, Western , Caspase 3 , Caspase 8 , Caspase 9 , Caspase Inhibitors , Cell Survival/radiation effects , Cells, Cultured/metabolism , Dose-Response Relationship, Radiation , Enzyme Activation/radiation effects , Enzyme Inhibitors/radiation effects , Epidermis/metabolism , Epidermis/radiation effects , Genes, bcl-2 , Genetic Vectors , Histocytochemistry , Humans , In Vitro Techniques , Keratinocytes/enzymology , Keratinocytes/metabolism , Keratinocytes/radiation effects , Mice , Mice, Inbred BALB C , Proto-Oncogene Proteins c-bcl-2/genetics , Proto-Oncogene Proteins c-bcl-2/metabolism , Skin/cytology , Skin/enzymology , Skin/metabolism , Skin/radiation effects , Transfection
19.
J Neurosci ; 21(16): 6105-14, 2001 Aug 15.
Article in English | MEDLINE | ID: mdl-11487634

ABSTRACT

Hippocampal pyramidal neurons in culture showed a developmental shift in synapse distribution from dendritic shafts to spines. Using dual wavelength time-lapse fluorescence microscopy, we analyzed the morphogenesis of three synaptic components: dendritic spines, postsynaptic densities (PSDs), and presynaptic vesicles. Local assembly of a major PSD protein, PSD-95, was spatially and temporally correlated with spine morphogenesis. Clustering of postsynaptic PSD-95 and that of a predominant synaptic vesicle protein, synaptophysin, were also correlated. In contrast, pre-existing PSD-95 clusters in dendritic shafts were preferentially eliminated without promoting spine formation. The local and stepwise assembly of synaptic components at the contact sites between dendritic protrusions and axons explains the developmental remodeling of excitatory synapses.


Subject(s)
Cell Surface Extensions/metabolism , Nerve Tissue Proteins/metabolism , Pyramidal Cells/metabolism , Synapses/metabolism , Synaptic Vesicles/metabolism , Adenoviridae/genetics , Aging/metabolism , Animals , Axons/metabolism , Axons/ultrastructure , Cell Differentiation , Cell Surface Extensions/ultrastructure , Cells, Cultured , Dendrites/metabolism , Dendrites/ultrastructure , Disks Large Homolog 4 Protein , Genes, Reporter , Guanylate Kinases , Hippocampus , Immunohistochemistry , Intracellular Signaling Peptides and Proteins , Luminescent Proteins/genetics , Membrane Proteins , Mice , Microscopy, Fluorescence , Nerve Tissue Proteins/genetics , Protein Transport/physiology , Pseudopodia/metabolism , Pseudopodia/ultrastructure , Pyramidal Cells/ultrastructure , Rats , Recombinant Fusion Proteins/genetics , Recombinant Fusion Proteins/metabolism , Synapses/ultrastructure , Synaptic Vesicles/ultrastructure , Synaptophysin/metabolism , Transfection
20.
Leuk Lymphoma ; 40(3-4): 351-6, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11426557

ABSTRACT

Multiple myeloma is characterized by the accumulation of malignant plasma cells in the bone marrow and rarely cured by chemotherapy. Villunger et al. showed that the neoplastic plasma cells express Fas ligand (FasL), which transmits a signal of apoptosis upon ligation to Fas, and suggested that the FasL suppresses the T-cells activated against malignant cells, resulting in escape from tumour immunity. We examined serum soluble FasL (sFasL) levels in 35 multiple myeloma patients to evaluate the correlation between sFasL levels and clinical characteristics. The serum sFasL levels were not affected by the disease status, serum monoclonal protein levels, or other prognostic factors. We could not determine whether the expression of FasL is involved in the poor clinical course of the disease.


Subject(s)
Membrane Glycoproteins/blood , Multiple Myeloma/blood , Adult , Aged , Aged, 80 and over , Blood Proteins , Fas Ligand Protein , Female , Humans , Immunoglobulins/blood , Lymphocyte Count , Male , Middle Aged , Multiple Myeloma/diagnosis , Myeloma Proteins/analysis , Myeloma Proteins/metabolism , Retrospective Studies , Severity of Illness Index , Solubility , Statistics, Nonparametric
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