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1.
J Laryngol Otol ; 137(7): 749-756, 2023 Jul.
Article in English | MEDLINE | ID: mdl-35916274

ABSTRACT

OBJECTIVE: For low-grade intraepithelial neoplasia cases, pharyngolaryngeal lesions equal to or less than 5 mm in size do not generally progress to invasive carcinoma. However, micro-superficial lesions equal to or less than 5 mm that showed rapid growth have been recently encountered. This study aimed to identify the characteristics of preferential progression of lesions equal to or less than 5 mm in size. METHOD: Gross findings, endoscopic findings and pathological results of 55 lesions measuring equal to or less than 5 mm in diameter were retrospectively reviewed to identify factors that distinguish squamous cell carcinoma or high-grade intraepithelial neoplasia from low-grade intraepithelial neoplasia or non-atypia lesions. RESULTS: The overall sensitivity, specificity, accuracy, and positive and negative predictive value of background colouration and intrapapillary capillary loop pattern in differentiation of squamous cell carcinoma or high-grade intraepithelial neoplasia from low-grade intraepithelial neoplasia or non-atypia lesions were all 100 per cent. CONCLUSION: Diagnosis based on background colouration and the intrapapillary capillary loop pattern on narrow-band imaging facilitates the pathological examination of lesions measuring equal to or less than 5 mm.


Subject(s)
Carcinoma in Situ , Carcinoma, Squamous Cell , Humans , Retrospective Studies , Carcinoma, Squamous Cell/diagnosis , Carcinoma, Squamous Cell/pathology , Narrow Band Imaging/methods , Predictive Value of Tests , Carcinoma in Situ/diagnostic imaging , Carcinoma in Situ/pathology
2.
Ultrasound Obstet Gynecol ; 58(6): 864-874, 2021 Dec.
Article in English | MEDLINE | ID: mdl-33942916

ABSTRACT

OBJECTIVES: To describe the prenatal neuroimaging spectrum of rhombencephalosynapsis (RES) and criteria for its classification according to the severity of vermian anomaly. METHODS: In this multicenter retrospective study of fetuses with RES between 2002 and 2020, the medical records and brain ultrasound and magnetic resonance images were evaluated comprehensively to determine the severity of the vermian anomaly and the presence of associated brain findings. RES was classified, according to the pattern of vermian agenesis and the extent of the fusion of the hemispheres, as complete RES (complete absence of the vermis) or partial RES (further classified according to the part of the vermis that was missing and, consequently, the region of hemispheric fusion, as anterior, posterior, severe or mixed RES). Findings were compared between cases with complete and those with partial RES. RESULTS: Included in the study were 62 fetuses with a gestational age ranging between 12 and 37 weeks. Most had complete absence of the vermis (complete RES, 77.4% of cases), a 'round-shaped' cerebellum on axial views (72.6%) and a transverse cerebellar diameter (TCD) < 3rd centile (87.1%). Among the 22.6% of cases with partial RES, 6.5% were classified as severe partial, 6.5% as partial anterior, 8.1% as partial mixed and 1.6% as partial posterior. Half of these cases presented with normal or nearly normal cerebellar morphology and 28.5% had a TCD within the normal limits. Infratentorially, the fourth ventricle was abnormal in 88.7% of cases overall, and anomalies of the midbrain and pons were frequent (93.5% and 77.4%, respectively). Ventriculomegaly was observed in 80.6% of all cases, being more severe in cases with complete RES than in those with partial RES, with high rates of parenchymal and septal disruption. CONCLUSIONS: This study provides prenatal neuroimaging criteria for the diagnosis and classification of RES, and identification of related features, using ultrasound and magnetic resonance imaging. According to our findings, a diagnosis of RES should be considered in fetuses with a small TCD (severe cerebellar hypoplasia) and/or a round-shaped cerebellum on axial views, during the second or third trimester, especially when associated with ventriculomegaly. Partial RES is more common than previously thought, but presents an extreme diagnostic challenge, especially in cases with normal or nearly-normal cerebellar morphobiometric features. © 2021 International Society of Ultrasound in Obstetrics and Gynecology.


Subject(s)
Abnormalities, Multiple/diagnostic imaging , Cerebellar Vermis/abnormalities , Cerebellum/abnormalities , Eye Abnormalities/diagnostic imaging , Kidney Diseases, Cystic/diagnostic imaging , Nervous System Malformations/diagnostic imaging , Neuroimaging , Prenatal Diagnosis/methods , Retina/abnormalities , Rhombencephalon/abnormalities , Abnormalities, Multiple/embryology , Adult , Cerebellar Vermis/diagnostic imaging , Cerebellar Vermis/embryology , Cerebellum/diagnostic imaging , Cerebellum/embryology , Eye Abnormalities/embryology , Female , Gestational Age , Humans , Kidney Diseases, Cystic/embryology , Magnetic Resonance Imaging , Multimodal Imaging , Nervous System Malformations/embryology , Pregnancy , Retina/diagnostic imaging , Retina/embryology , Retrospective Studies , Rhombencephalon/diagnostic imaging , Rhombencephalon/embryology , Severity of Illness Index , Ultrasonography, Prenatal
3.
J Laryngol Otol ; 135(9): 773-778, 2021 Sep.
Article in English | MEDLINE | ID: mdl-33928889

ABSTRACT

OBJECTIVE: Severe acute respiratory syndrome coronavirus-2 uses angiotensin-converting enzyme-2 as a primary receptor for invasion. This study investigated angiotensin-converting enzyme-2 expression in the sinonasal mucosa of patients with chronic rhinosinusitis, as this could be linked to a susceptibility to severe acute respiratory syndrome coronavirus-2 infection. METHODS: Ethmoid sinus specimens were obtained from 27 patients with eosinophilic chronic rhinosinusitis, 18 with non-eosinophilic chronic rhinosinusitis and 18 controls. The angiotensin-converting enzyme-2 and other inflammatory cytokine and chemokine messenger RNA levels were assessed by quantitative reverse transcription polymerase chain reaction. Angiotensin-converting enzyme-2 positive cells were examined immunohistologically. RESULTS: The eosinophilic chronic rhinosinusitis patients showed a significant decrease in angiotensin-converting enzyme-2 messenger RNA expression. In the chronic rhinosinusitis patients, angiotensin-converting enzyme-2 messenger RNA levels were positively correlated with tumour necrosis factor-α and interleukin-1ß (r = 0.4971 and r = 0.3082, respectively), and negatively correlated with eotaxin-3 (r = -0.2938). Angiotensin-converting enzyme-2 immunoreactivity was mainly localised in the ciliated epithelial cells. CONCLUSION: Eosinophilic chronic rhinosinusitis patients with type 2 inflammation showed decreased angiotensin-converting enzyme-2 expression in their sinus mucosa. Angiotensin-converting enzyme-2 regulation was positively related to pro-inflammatory cytokines, especially tumour necrosis factor-α production, in chronic rhinosinusitis patients.


Subject(s)
Angiotensin-Converting Enzyme 2/metabolism , Nasal Mucosa/enzymology , Rhinitis/enzymology , Sinusitis/enzymology , Adult , COVID-19/etiology , Chronic Disease , Cross-Sectional Studies , Female , Humans , Male , Middle Aged , Nasal Mucosa/metabolism , Nasal Mucosa/virology , Reverse Transcriptase Polymerase Chain Reaction , Rhinitis/complications , Rhinitis/metabolism , SARS-CoV-2/metabolism , Sinusitis/complications , Sinusitis/metabolism
4.
J Hosp Infect ; 103(4): 395-403, 2019 Dec.
Article in English | MEDLINE | ID: mdl-31425718

ABSTRACT

BACKGROUND: Outbreaks of vancomycin-resistant enterococcus (VRE) are a serious problem in hospitals. Inferring the transmission route is an important factor to institute appropriate infection control measures; however, the methodology has not been fully established. AIM: To reconstruct and evaluate the transmission model using sequence variants extracted from whole genome sequencing (WGS) data and epidemiological information from patients involved in a VRE outbreak. METHODS: During a VRE outbreak in our hospital, 23 samples were collected from patients and environmental surfaces and analysed using WGS. By combining genome alignment information with patient epidemiological data, the VRE transmission route was reconstructed using a Bayesian approach. With the transmission model, evaluation and further analyses were performed to identify risk factors that contributed to the outbreak. FINDINGS: All VREs were identified as Enterococcus faecium belonging to sequence type 17, which consisted of two VRE genotypes: vanA (N = 8, including one environmental sample) and vanB (N = 15). The reconstruction model using the Bayesian approach showed the transmission direction with posterior probability and revealed transmission through an environmental surface. In addition, some cases acting as VRE spreaders were identified, which can interfere with appropriate infection control. Vancomycin administration was identified as a significant risk factor for spreaders. CONCLUSION: A Bayesian approach for transmission route reconstruction using epidemiologic data and genomic variants from WGS can be applied in actual VRE outbreaks. This may contribute to the design and implementation of effective infection control measures.


Subject(s)
Disease Transmission, Infectious , Enterococcus faecium/isolation & purification , Gram-Positive Bacterial Infections/epidemiology , Gram-Positive Bacterial Infections/transmission , Molecular Epidemiology , Vancomycin-Resistant Enterococci/isolation & purification , Whole Genome Sequencing , Adult , Aged , Aged, 80 and over , Bayes Theorem , Cross Infection/epidemiology , Cross Infection/transmission , Disease Outbreaks , Enterococcus faecium/classification , Enterococcus faecium/genetics , Female , Genotype , Humans , Male , Middle Aged , Molecular Typing , Risk Factors , Vancomycin-Resistant Enterococci/classification , Vancomycin-Resistant Enterococci/genetics
5.
J Clin Pharm Ther ; 43(6): 799-806, 2018 Dec.
Article in English | MEDLINE | ID: mdl-29770467

ABSTRACT

WHAT IS KNOWN AND OBJECTIVE: Anticholinergic drugs are associated with risks of falls, confusion and cognitive dysfunction. However, the effect of anticholinergic drug use on rehabilitation outcomes after a stroke is poorly documented. We therefore aimed to establish whether the anticholinergic load was associated with functional recovery among geriatric patients convalescing after stroke. METHOD: Consecutive geriatric stroke patients admitted and discharged from a convalescence rehabilitation ward between 2010 and 2016 were included in this retrospective cohort study. Anticholinergic load was assessed by the Anticholinergic Risk Scale (ARS), and functional recovery was assessed by the Functional Independence Measure (FIM). The primary outcome was cognitive FIM (FIM-C) gain, but we also assessed the interaction of other putative factors identified from univariate analysis. Multivariate analyses were performed, adjusting for confounding factors. RESULTS AND DISCUSSION: We included 418 participants (171 males, 247 females) with a median age of 78 years (interquartile range, 72-84 years). Multiple regression analysis revealed that ARS change, length of stay, and epilepsy were independently and negatively correlated with cognitive FIM gain. Multiple logistic regression analysis indicated that the "Comprehension" and "Memory" items of the cognitive FIM gain were independently and negatively associated with anticholinergic load. WHAT IS NEW AND CONCLUSION: A causal relationship cannot be established, but increased ARS scores during hospitalization may predict limited cognitive functional improvement in geriatric patients after stroke. Alternatively, cognitive impairment may lead to increased use of anticholinergic drugs.


Subject(s)
Activities of Daily Living/psychology , Cholinergic Antagonists/administration & dosage , Cognition/drug effects , Stroke/therapy , Aged , Aged, 80 and over , Cholinergic Antagonists/adverse effects , Cognition/physiology , Cohort Studies , Female , Hospitalization , Humans , Length of Stay , Male , Multivariate Analysis , Recovery of Function/drug effects , Regression Analysis , Retrospective Studies , Stroke/psychology , Stroke Rehabilitation/methods , Treatment Outcome
6.
Rhinology ; 47(3): 242-247, 2009 09.
Article in English | MEDLINE | ID: mdl-19839244

ABSTRACT

We investigated the expression of transient receptor potential vanilloid (TRPV)-1-4 in the mouse olfactory epithelium (OE) in comparison to its expression in respiratory epithelium. CBA/J mice were used. The localization of TRPV-1, -2, -3 and -4 in the nasal mucosa was investigated using immunohistochemistry and a double staining study for TRPV-1 and -2 and SP was also carried out. TRPV-1-4, were expressed variably in the OE with a diffuse pattern in lamina propria, and were expressed in respiratory epithelium with strong positive expression in glandular cells of lamina propria. The double-staining study revealed coexpression of TRPV-1 and -2 and substance P (SP) in the trigeminal nerve fibers of the OE. Coexpression of TRPV-1 and SP was marked around the blood vessels and seromucinous gland of respiratory epithelium while TRPV-2 showed no co-localization. TRPV-1-4 were found to be localized in the mouse OE and respiratory epithelium. Our results suggest that TRPVs may play multiple roles in the OE, contributing to olfactory adaptation, olfactory/trigeminal interactions in nasal chemoreception and OE homeostasis; they may also be involved in olfactory transduction as well as olfactory dysfunction secondary to sinonasal inflammatory disease. TRPVs in respiratory mucosa may play a significant role in nasal nociception, ciliary movement and the regulation of mucous secretion.


Subject(s)
Olfactory Mucosa/metabolism , TRPV Cation Channels/metabolism , Animals , Calcium Channels/metabolism , Immunohistochemistry , Mice , Mice, Inbred CBA , Nasal Mucosa/metabolism , Respiratory Mucosa/metabolism
7.
Res Vet Sci ; 87(3): 349-54, 2009 Dec.
Article in English | MEDLINE | ID: mdl-19535119

ABSTRACT

Cyclooxygenase (COX)-2 is an inducible isoform of COX and is expressed under abnormal health conditions. This study elucidated the cutaneous induction of COX-2 during the wound healing processes in dog skin. Dog skin was sutured after punch biopsy and investigated histologically and immunohistochemically on days 0 (normal), 1, 3, 5, 7, 10, and 14 after injury. Histological changes, including infiltration of inflammatory cells and proliferation of fibroblast-like cells, were observed as predicted, and there was a close and significant correlation between these 2 events. COX-2-positive cells were detected in the epidermis between days 1 and 7, and bimodal peaks were observed in the case of the percentage of COX-2-positive cells. In inflammatory cells, COX-positive signals were detected on day 3 only. Here, we clarified the localization and pattern of the induced COX-2 expression during wound healing in dog skin.


Subject(s)
Cyclooxygenase 2/metabolism , Dogs/injuries , Dogs/physiology , Gene Expression Regulation, Enzymologic/physiology , Skin/enzymology , Wound Healing/physiology , Animals , Cyclooxygenase 2/genetics , Immunohistochemistry/veterinary , Protein Transport , Time Factors
8.
J Phys Chem A ; 113(13): 3143-54, 2009 Apr 02.
Article in English | MEDLINE | ID: mdl-19320516

ABSTRACT

Raman spectra of the C[triple bond]N stretching vibration of p-aminobenzonitrile (ABN) have been investigated in water, methanol, and cyclohexane under sub- and supercritical conditions, and in acetonitrile under subcritical condition. In all solvent fluids covering the supercritical region, the vibrational frequency of the C[triple bond]N stretching mode decreased with increasing solvent density from the gaseous region to the medium density region rho(r) approximately = 2, where rho(r) is the reduced density by the critical density of the solvent. However, from the medium density region to the higher density region, the vibrational frequency turned to increase with the solvent density. The temperature-induced low frequency shift of the C[triple bond]N stretching Raman band was also ascertained by the measurement of the temperature dependence of Raman spectrum of ABN vapor above 543 K. The electronic absorption spectra in the UV region of ABN were also measured under the same experimental conditions. The absorption peak energies decreased with an increase of the solvent density, except in water above rho(r) = 2.8. The vibrational frequency shift in cyclohexane was explained by a sum of contributions of the repulsive interaction, the mean field attractive interaction, and the pure temperature effect probably due to the hot-band contribution. The residual frequency shift after the subtraction of the repulsive and temperature effects in water and methanol showed the low frequency shift with increasing solvent density from rho(r) congruent with 0 to 2.8. However, above rho(r) congruent with 2.8 in water, the residual shift showed a high frequency shift with increasing solvent density. The electronic state calculations based on the PCM model using the density functional theory (DFT) indicated that the solvent polarity change caused the low frequency shift of the C[triple bond]N stretching mode, which was also correlated with the shift of the electronic absorption spectrum. The results of the DFT calculations on the cluster of ABN with water molecules and the molecular dynamics simulations indicated that the high frequency shift of the C[triple bond]N stretching mode in water above rho(r) congruent with 2.8 could be due to the hydrogen bonding between water and ABN.

9.
J Thromb Haemost ; 7(1): 111-20, 2009 Jan.
Article in English | MEDLINE | ID: mdl-19017259

ABSTRACT

BACKGROUND: Tissue factor pathway inhibitor (TFPI) is a physiological protease inhibitor that inhibits the initial reactions of the extrinsic blood coagulation pathway. Most TFPI in human plasma is associated with lipoproteins; however, the most functionally active form is thought to be the free, full-length form (f-pTFPI). Cell culture derived TFPI and recombinant TFPI (rTFPI) exhibit variations in their respective anticoagulant activity, which may be caused by post-translational modifications, such as the frequent differences in sugar chain structures among recombinant proteins. Sugar chain structures in rTFPI expressed in Chinese hamster ovary (CHO) cells have been reported previously, but those of plasma TFPI have not been. OBJECTIVES: To purify f-pTFPI and analyze the sugar chain structures. RESULTS AND CONCLUSION: f-pTFPI was purified to homogeneity from blood plasma using a combination of anion-exchange, heparin affinity, immunoaffinity, and reversed-phase chromatographies, resulting in a yield of 76%. f-pTFPI showed a partially phosphorylated glycoprotein comprising a total of 276 amino acids by peptide mapping. The sugar chain structures were analyzed by two-dimensional sugar mapping combined with exoglycosidase digestion of the pyridylamino sugar chains and the following results were obtained. (Sialyl) Galbeta1-3GalNAc was linked to Thr(175), partially to Thr(14) and Ser(174); sialyl complex-type sugar chains to Asn(117) and Asn(167), whereas Asn(228) was not glycosylated. Neuraminidase-resistant acidic sugar chains including sulfated sugar chains were not observed significantly. The protease inhibitory activities of f-pTFPI towards activated factor (F) X and tissue factor-activated FVII complex were identical to those of full-length rTFPI expressed in CHO cells.


Subject(s)
Carbohydrates/analysis , Lipoproteins/chemistry , Protein Processing, Post-Translational , Chromatography , Glycoproteins/analysis , Humans , Lipoproteins/blood , Lipoproteins/isolation & purification , Phosphorylation , Protease Inhibitors
10.
Clin Exp Immunol ; 150(3): 397-406, 2007 Dec.
Article in English | MEDLINE | ID: mdl-17900307

ABSTRACT

Intravenous immunoglobulin (IVIg) preparations are reportedly effective in inhibiting the relapse of multiple sclerosis (MS), but few reports have investigated the effect of IVIg on dendritic cells (DCs), which are thought to be involved in such relapses. In the system that uses monokines to differentiate DCs from peripheral blood monocytes (Mo-DCs), we investigated the effect of immunoglobulin G (IgG) on these antigen-presenting cells. Using monocytes derived from healthy volunteers, IgG partially inhibited the expression of CD1a, a marker of immature DCs (imDCs), and CD40 and CD80, which are markers associated with T cell activation. In contrast, IgG enhanced the expression of CD83, a marker of mature DCs (mDCs). Furthermore, IgG markedly inhibited the expression of CD49d [very late activation antigen (VLA)-4 alpha4-integrin], the adhesion molecule required for mDCs to cross the blood-brain barrier. We obtained similar results on all the aforementioned cell surface molecules investigated in both healthy controls and MS patients. In addition, IgG treatment of cells from both healthy controls and MS patients inhibited the production of interleukin (IL)-12, a cytokine associated with mDC differentiation, but did not inhibit the production of IL-10. These results suggested the possibility that IgG treatment, apart from its known ability to regulate inflammation, may help to prevent relapses of MS by controlling DC maturation, consequently inhibiting invasion of immune cells into the central nervous system and affecting the cytokine profile.


Subject(s)
Dendritic Cells/immunology , Immunoglobulin G/immunology , Multiple Sclerosis, Relapsing-Remitting/immunology , Adult , Antigens, CD/metabolism , Cell Differentiation/immunology , Cells, Cultured , Dendritic Cells/cytology , Female , Humans , Immunoglobulins, Intravenous/immunology , Integrin alpha4/metabolism , Interleukin-10/biosynthesis , Interleukin-12/biosynthesis , Male , Middle Aged , Monocytes/cytology , Monocytes/immunology
11.
Int J Hyperthermia ; 23(4): 363-70, 2007 Jun.
Article in English | MEDLINE | ID: mdl-17558735

ABSTRACT

Augmented expression of members of the heat shock protein 70 (Hsp70) family are frequently observed in various human cancers. In this study, we examined applicability of laser scanning cytometer (LSC) to evaluate the level of Hsp72, which is the member constitutively expressed and significantly induced after heat shock, in human tumour cell lines. The relative nuclear content of Hsp72 measured by LSC correlated well with the relative intracellular content determined by Western blotting (R = 0.906). Furthermore, there was a close relationship between the relative nuclear content of Hsp72 measured by LSC and the colony-forming ability in soft agar, one of the malignant characteristics of tumour cells (R = 0.880). These results indicate that LSC measurement is useful for predicting the degree of malignancy of cancer cells, as it is reliable, faster than Western blotting and more objective and quantitative than visual measurements.


Subject(s)
HSP72 Heat-Shock Proteins/metabolism , HSP72 Heat-Shock Proteins/radiation effects , Hot Temperature , Neoplasms/metabolism , Phenotype , Blotting, Western , Cell Line, Tumor , Humans , Hyperthermia, Induced , Laser Scanning Cytometry , Predictive Value of Tests
12.
Inflammopharmacology ; 15(1): 1-4, 2007 Feb.
Article in English | MEDLINE | ID: mdl-17323186

ABSTRACT

COX (cyclooxygenase) is one of the key enzymes involved in the synthesis of a variety of prostaglandins (PGs), some of which have been strongly linked to inflammation. One of its two well-known isoforms, COX-2, is an inducible enzyme whose induction and expression is dynamically regulated by growth factors, mitogens, and tumor promoters. Several animal and clinical studies have reported the chemopreventive effect of celecoxib, a selective COX-2 inhibitor; and in particular, a few studies have shown that celecoxib prevents the development of gastric cancer. Administration of celecoxib also showed increases in cardiovascular risk and disruption of renal physiology. Therefore, studies hoping to clarify how selective COX-2 inhibitors modulate gastric cancer must keep in mind that coxibs have also been linked to serious cardiovascular events and disruption of renal physiology.


Subject(s)
Cyclooxygenase 2 Inhibitors/adverse effects , Cyclooxygenase 2 Inhibitors/therapeutic use , Pyrazoles/adverse effects , Pyrazoles/therapeutic use , Stomach Neoplasms/prevention & control , Sulfonamides/adverse effects , Sulfonamides/therapeutic use , Animals , Cardiovascular Diseases/chemically induced , Celecoxib , Cyclooxygenase 2/drug effects , Cyclooxygenase 2 Inhibitors/pharmacology , Gastric Mucosa/drug effects , Helicobacter pylori/drug effects , Humans , Kidney/drug effects , Kidney/physiology , Metaplasia , Pyrazoles/pharmacology , Stomach Neoplasms/pathology , Sulfonamides/pharmacology
13.
Amino Acids ; 32(1): 79-85, 2007 Jan.
Article in English | MEDLINE | ID: mdl-17021656

ABSTRACT

The accumulation of D-isomers of aspartic acid (D-Asp) in proteins during aging has been implicated in the pathogenesis of Alzheimer's disease (AD), cataracts and arteriosclerosis. Here, we identified a specific lactacystin-sensitive endopeptidase that cleaves the D-Asp-containing protein and named it D-aspartyl endopeptidase (DAEP). DAEP has a multi-complex structure (MW: 600 kDa) and is localized in the inner mitochondrial membrane. However, DAEP activity was not detected in E. coli, S. cerevisiae, and C. elegans. A specific inhibitor for DAEP, i-DAEP: (benzoyl-L-Arg-L-His-[D-Asp]-CH(2)Cl; MW: 563.01), was newly synthesized and inhibited DAEP activity (IC(50), 3 microM), a factor of ten greater than lactacystin on DAEP. On the other hand, i-DAEP did not inhibit either the 20S or 26S proteasome. And we identified succinate dehydrogenase and glutamate dehydrogenase 1 as components of DAEP by affinity label using biotinylated i-DAEP. In the long life span of mammals, DAEP may serve as a scavenger against accumulation of racemized proteins in aging. Insights into DAEP will provide the foundation for developing treatments of diseases, such as AD, in which accumulation of D-Asp-containing proteins are implicated.


Subject(s)
Alzheimer Disease/enzymology , Aspartic Acid Endopeptidases/chemistry , D-Aspartic Acid/chemistry , Mitochondrial Membranes/enzymology , Mitochondrial Proteins/chemistry , Multienzyme Complexes/chemistry , Oligopeptides/chemistry , Protease Inhibitors/chemistry , Acetylcysteine/analogs & derivatives , Acetylcysteine/chemistry , Aging/metabolism , Alzheimer Disease/drug therapy , Animals , Aspartic Acid Endopeptidases/antagonists & inhibitors , Aspartic Acid Endopeptidases/isolation & purification , Caenorhabditis elegans/enzymology , D-Aspartic Acid/metabolism , Escherichia coli/enzymology , Glutamate Dehydrogenase/antagonists & inhibitors , Glutamate Dehydrogenase/chemistry , Glutamate Dehydrogenase/isolation & purification , Mice , Mitochondrial Proteins/antagonists & inhibitors , Mitochondrial Proteins/isolation & purification , Multienzyme Complexes/antagonists & inhibitors , Multienzyme Complexes/isolation & purification , Oligopeptides/chemical synthesis , Oligopeptides/therapeutic use , Protease Inhibitors/chemical synthesis , Protease Inhibitors/therapeutic use , Proteasome Endopeptidase Complex/chemistry , Proteasome Inhibitors , Rabbits , Saccharomyces cerevisiae/enzymology , Succinate Dehydrogenase/antagonists & inhibitors , Succinate Dehydrogenase/chemistry , Succinate Dehydrogenase/isolation & purification
14.
Dig Dis Sci ; 50(12): 2323-9, 2005 Dec.
Article in English | MEDLINE | ID: mdl-16416182

ABSTRACT

Rebamipide has a broad spectrum of pharmacological actions that include suppression of neutrophil functions and stimulation of mucosal epithelial cell regeneration by increasing the expression of epithelial growth factor (EGF) and the EGF receptor. Sixteen patients with active ulcerative colitis (UC; mild in 1 patient, moderate in 11, and severe in 4) were recruited. Enemas containing 150 mg rebamipide per dosing were administered during the daytime after passage of stool, twice a day for 4 weeks. UC disease activity index (UC-DAI), endoscopic activity index (EAI), and Floren's grading (FG) of mucosal biopsy specimens were measured at entry and at 4 weeks. Five of 16 patients did not complete the study, and therefore, final efficacy assessment was done on 11 patients who completed the 4 weeks of treatment. Improvements were observed in UC-DAI (P = 0.0049), EAI (P = 0.0043), and FG (P = 0.0084). There was no serious rebamipide-related side effect in any of the 16 patients. In conclusion, rebamipide topical therapy appears to be effective for the treatment of mildly to moderately active distal UC. Further controlled studies are warranted for this promising drug.


Subject(s)
Anti-Ulcer Agents/administration & dosage , Colitis, Ulcerative/drug therapy , Enema , Proctitis/drug therapy , Adolescent , Adult , Aged , Colitis, Ulcerative/diagnosis , Dose-Response Relationship, Drug , Endoscopy, Gastrointestinal , Female , Follow-Up Studies , Humans , Intestinal Mucosa/drug effects , Intestinal Mucosa/pathology , Male , Middle Aged , Patient Compliance , Probability , Proctitis/diagnosis , Prospective Studies , Risk Assessment , Severity of Illness Index , Statistics, Nonparametric , Treatment Outcome
15.
Biosci Biotechnol Biochem ; 65(8): 1736-40, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11577711

ABSTRACT

A novel method for synthesizing CMP-NeuAc was established. We first confirmed that the putative neuA gene of Haemophilus influenzae, identified by its whole genome sequence project, indeed encodes CMP-NeuAc synthetase (EC 2.7.7.43). The enzyme requires CTP as a cytidylyl donor for cytidylylation of NeuAc. The enzyme was coupled with an enzymatic CTP-generating system from CMP and inorganic polyphosphate as a sole phospho-donor driven by the combination of polyphosphate kinase and CMP kinase, where phosphorylation of CMP is done by the combined activity expressed by both enzymes, and subsequent phosphorylation of CDP by polyphosphate kinase itself occurred efficiently. When CMP-NeuAc synthetase of H. influenzae, polyphosphate kinase, and CMP kinase were added to the reaction mixture containing equimolar concentrations (15 mM) of CMP and NeuAc, and polyphosphate (150 mM in terms of phosphate), CMP-NeuAc was synthesized up to 10 mM in 67% yield.


Subject(s)
Cytidine Monophosphate/analogs & derivatives , Cytidine Monophosphate/chemical synthesis , N-Acylneuraminate Cytidylyltransferase/chemical synthesis , Sialic Acids/chemical synthesis , Cytidine Triphosphate/chemical synthesis , Escherichia coli/genetics , Escherichia coli/metabolism , Haemophilus influenzae/enzymology , Haemophilus influenzae/genetics , Phosphotransferases (Phosphate Group Acceptor)/chemistry , Phosphotransferases (Phosphate Group Acceptor)/genetics , Plasmids/genetics , Recombinant Proteins/chemical synthesis
16.
J Biochem ; 130(4): 475-80, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11574066

ABSTRACT

We identified a novel metalloprotease, which could be responsible for cleaving the Tyr842-Met843 peptide bond of von Willebrand factor (vWF). This metalloprotease was purified from Cohn Fraction-I precipitate of human pooled plasma by the combination of gel filtration, DEAE chromatography, and preparative polyacrylamide gel electrophoresis in the presence of SDS. The NH2-terminal amino acid sequence of the isolated protein was: AAGGILHLELLVAVGPDVFQAHQEDTRRY. Based on this sequence, we searched human genomic and EST databases, and identified compatible nucleotide sequences. These results suggested that this protein is a novel metalloprotease, a member of the family of a disintegrin and metalloprotease with thrombospondin type-1 motifs (ADAMTS), and its genomic DNA was mapped to human chromosome 9q34. Multiple human tissue northern blotting analysis indicated that the mRNA encoding this protease spanned approximately 5 kilobases and was uniquely expressed in the liver. Furthermore, we determined the cDNA sequence encoding this protease, and found that this protease was comprised of a signal peptide, a proregion followed by the putative furin cleavage site, a reprolysin-type zinc-metalloprotease domain, a disintegrin-like domain, a thrombospondin type-1 (TSP1) motif, a cysteine-rich region, a spacer domain, and COOH-terminal TSP1 motif repeats.


Subject(s)
Liver/enzymology , Metalloendopeptidases/biosynthesis , von Willebrand Factor/metabolism , ADAM Proteins , ADAMTS13 Protein , Alternative Splicing , Amino Acid Motifs , Amino Acid Sequence , Base Sequence , Chromosome Mapping , Cloning, Molecular , Disintegrins/chemistry , Humans , Metalloendopeptidases/genetics , Metalloendopeptidases/metabolism , Molecular Sequence Data , Protein Isoforms/genetics , Protein Structure, Tertiary , RNA, Messenger/biosynthesis , Thrombospondin 1/chemistry , Tissue Distribution , Transcription, Genetic
17.
J Histochem Cytochem ; 49(9): 1123-31, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11511681

ABSTRACT

Two types of NADP-dependent isocitrate dehydrogenases (ICDs) have been reported: mitochondrial (ICD1) and cytosolic (ICD2). The C-terminal amino acid sequence of ICD2 has a tripeptide peroxisome targeting signal 1 sequence (PTS1). After differential centrifugation of the postnuclear fraction of rat liver homogenate, approximately 75% of ICD activity was found in the cytosolic fraction. To elucidate the true localization of ICD2 in rat hepatocytes, we analyzed the distribution of ICD activity and immunoreactivity in fractions isolated by Nycodenz gradient centrifugation and immunocytochemical localization of ICD2 antigenic sites in the cells. On Nycodenz gradient centrifugation of the light mitochondrial fraction, ICD2 activity was distributed in the fractions in which activity of catalase, a peroxisomal marker, was also detected, but a low level of activity was also detected in the fractions containing activity for succinate cytochrome C reductase (a mitochondrial marker) and acid phosphatase (a lysosomal marker). We have purified ICD2 from rat liver homogenate and raised a specific antibody to the enzyme. On SDS-PAGE, a single band with a molecular mass of 47 kD was observed, and on immunoblotting analysis of rat liver homogenate a single signal was detected. Double staining of catalase and ICD2 in rat liver revealed co-localization of both enzymes in the same cytoplasmic granules. Immunoelectron microscopy revealed gold particles with antigenic sites of ICD2 present mainly in peroxisomes. The results clearly indicated that ICD2 is a peroxisomal enzyme in rat hepatocytes. ICD2 has been regarded as a cytosolic enzyme, probably because the enzyme easily leaks out of peroxisomes during homogenization. (J Histochem Cytochem 49:1123-1131, 2001)


Subject(s)
Cytosol/enzymology , Isocitrate Dehydrogenase/metabolism , Liver/enzymology , NADP/metabolism , Peroxisomes/enzymology , Animals , Antibody Specificity , Catalase/metabolism , Cell Fractionation , Centrifugation, Density Gradient , Fluorescent Antibody Technique , Frozen Sections , Immunoblotting , Immunohistochemistry , Isocitrate Dehydrogenase/immunology , Liver/ultrastructure , Male , Microscopy, Electron/methods , Organelles/enzymology , Rats , Rats, Wistar
18.
Oncol Rep ; 8(5): 973-80, 2001.
Article in English | MEDLINE | ID: mdl-11496301

ABSTRACT

Nerve growth factor (NGF) is a member of the neurotrophin family and is essential for the differentiation and maintenance of neural cells. Recently, it has been reported that NGF is involved in the growth of breast cancer. On the other hand, two types of NGF receptors have been identified, a low-affinity receptor, p75NGFR, and a high-affinity receptor, TrkA. NGF-p75NGFR interaction is known to play an important role in apoptosis, whereas NGF-TrkA interaction is responsible for the survival of neural cells. We examined the relationship between clinicopathological factors, Ki-67 index, apoptotic index and the immunohistochemical expression of NGF, TrkA and p75NGFR in 71 invasive ductal breast carcinoma (IDBC) specimens. Our data indicate that positive Ki-67 expression (a labeling index exceeding 30%) correlates significantly with the positive expression of NGF (p=0.0091). Moreover, the apoptotic index was found to correlate with a strong expression of p75NGFR. Furthermore, patients who were NGF positive and p75NGFR negative had significantly poorer disease-free survival rates (p=0.0165). In contrast, those who were NGF negative and p75NGFR positive had significantly more favorable outcomes (p=0.0191). These findings suggest that a combined evaluation of NGF and p75NGFR expression is a predictive factor in the prognosis of IDBC patients.


Subject(s)
Biomarkers, Tumor/metabolism , Breast Neoplasms/metabolism , Carcinoma, Ductal, Breast/metabolism , Nerve Growth Factors/metabolism , Receptors, Nerve Growth Factor/metabolism , Adult , Aged , Aged, 80 and over , Breast Neoplasms/diagnosis , Breast Neoplasms/mortality , Carcinoma, Ductal, Breast/diagnosis , Carcinoma, Ductal, Breast/mortality , Cell Division/physiology , Disease-Free Survival , Female , Humans , Immunoenzyme Techniques , Middle Aged , Mitotic Index , Multivariate Analysis , Prognosis , Receptor, Nerve Growth Factor , Receptor, trkA/metabolism , Survival Rate
19.
Pharmacology ; 62(1): 10-6, 2001 Jan.
Article in English | MEDLINE | ID: mdl-11150917

ABSTRACT

Nitric oxide synthase (NOS) catalyzes nitric oxide (NO) formation from L-arginine in the presence of molecular oxygen and NADPH. NO is involved in the regulation of microvasculature. Isosorbide dinitrate (ISDN) and glyceryl trinitrate (GTN) have been widely used as vasodilators to treat acute myocardial ischemia, their biological effects being due to the release of NO. In this investigation, the effects of ISDN and GTN on NOS activity in the presence or absence of oxyhemoglobin under hypoxia and normoxia were studied. The apparent K(m) values for molecular oxygen were 21.6 +/- 1.5 and 9.4 +/- 1.3 micromol/l for nNOS and eNOS, respectively. ISDN liberated NO in a concentration- and pH-dependent manner, but no differences between hypoxia and normoxia were observed. The NO release from ISDN was also measured directly by an electron spin resonance spectral method with N-(dithiocarboxy)sarcosine-Fe complex as a NO-trapping agent. ISDN increased nNOS and eNOS activities in the presence of 30 micromol/l oxyhemoglobin under hypoxia, while it did not affect nNOS and eNOS activities under normoxia. In the absence of oxyhemoglobin, ISDN inhibited nNOS and eNOS activities under both hypoxic and normoxic experimental conditions. The rate of oxygen release from oxyhemoglobin under hypoxia was increased 3 times in the presence of 1 mmol/l ISDN. In contrast to ISDN, GTN could not release NO spontaneously, and it also did not affect nNOS and eNOS activities in the absence or presence of 30 micromol/l oxyhemoglobin under both hypoxic and normoxic conditions. These results indicated that the NO release from ISDN is different from that of GTN, and the increase of NOS activity by ISDN in the presence of oxyhemoglobin under hypoxia is ascribed to the increase in molecular oxygen concentration.


Subject(s)
Isosorbide Dinitrate/pharmacology , Nitric Oxide Synthase/drug effects , Oxygen/pharmacology , Animals , Citrulline/drug effects , Citrulline/metabolism , Dose-Response Relationship, Drug , Hydrogen-Ion Concentration , Kinetics , Nitric Oxide/metabolism , Nitric Oxide Synthase/metabolism , Nitroglycerin/pharmacology , Oxyhemoglobins/pharmacology , Swine
20.
Biochem J ; 353(Pt 2): 325-31, 2001 Jan 15.
Article in English | MEDLINE | ID: mdl-11139397

ABSTRACT

The effect of two arginine-specific cysteine proteinases (gingipains R) from Porphyromonas gingivalis, an aetiological factor of adult periodontitis, on the activation of human factor IX was investigated in the presence of ethylene glycol, an activity enhancer of activated factor IX (factor IXa), with the use of a fluorogenic oligopeptide substrate. Each gingipain R rapidly activated factor IX but the 95 kDa proteinase complex (HRgpA) that contains both haemagglutinin/adhesion and catalytic domains was 2.4-fold more efficient than the single-chain 50 kDa gingipain R (RgpB), which has only a catalytic domain. SDS/PAGE and N-terminal sequence analysis of factor IX digestion fragments indicated that, like all endogenous activators, gingipains R also produce factor IXabeta via an IXa intermediate. Significantly, phospholipids augmented the activation of factor IX by HRgpA but not by RgpB in a Ca(2+)-dependent manner. In the presence of both cofactors the kinetic efficiency of HRgpA to activate factor IX (k(cat)/K(m)=1.9x10(6) M(-1).s(-1)) was 8.5-fold higher than that of RgpB (k(cat)/K(m)=2.3x10(5) M(-1).s(-1)) and double that of the factor VIIa-tissue factor complex, but 8-fold lower than that for factor XIa. A comparison of the relative activation rates of factor IX, factor X and prothrombin directly in plasma by HRgpA suggests a significant contribution for factor IX conversion in blood coagulation induced by gingipains R. Taken together, gingipains R are the first-reported activators of factor IX of bacterial origin. By this effect they could be involved in the production of thrombin as well as the subsequent generation of prostaglandins and interleukin 1, all of which have been found to be associated with the development and progression of periodontitis.


Subject(s)
Bacterial Proteins/pharmacology , Cysteine Endopeptidases/pharmacology , Factor IX/metabolism , Hemagglutinins/pharmacology , Porphyromonas gingivalis/enzymology , Adhesins, Bacterial , Cysteine Endopeptidases/isolation & purification , Factor IXa/metabolism , Factor X/metabolism , Gingipain Cysteine Endopeptidases , Hemagglutinins/isolation & purification , Humans , Isoenzymes/pharmacology , Kinetics , Phospholipids/pharmacology , Porphyromonas gingivalis/chemistry , Prothrombin/metabolism
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