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1.
Endoscopy ; 45(3): 195-201, 2013.
Article in English | MEDLINE | ID: mdl-23299524

ABSTRACT

BACKGROUND AND STUDY AIMS: The aim of the current study was to assess the detection rate of the right adrenal gland and the diagnostic ability of endoscopic ultrasound (EUS) and fine-needle aspiration (FNA) for the diagnosis of adrenal metastasis in potentially resectable lung cancer. PATIENTS AND METHODS: This retrospective cohort study included a consecutive series of 150 patients undergoing EUS/EUS - FNA for staging of lung cancer. The detection rate of the right adrenal gland by EUS and the diagnostic accuracies of computed tomography (CT), positron emission tomography-CT (PET-CT), and EUS/EUS - FNA for the diagnosis of adrenal metastasis were evaluated. RESULTS: The right adrenal gland was visualized by EUS in 131 patients (87.3 %); the left adrenal gland was visualized in all patients. Findings suggestive of metastasis in either one of the adrenal glands or in both were observed in 6 patients (4.0 %) by CT, in 5 patients (3.3 %) by PET-CT, and in 11 patients (7.3 %) by EUS. EUS - FNA was performed simultaneously in the 11 patients, and in 4 patients the diagnosis of metastasis was established. The accuracy for the diagnosis of adrenal metastasis was 100 % for EUS/EUS - FNA, 96.0 % for CT, and 97.0 % for PET-CT (P = 0.1146). CONCLUSIONS: As well as the left adrenal gland, the right adrenal gland was also usually visible by EUS. EUS/EUS - FNA provided an accurate diagnosis of adrenal metastasis, although the prevalence of adrenal metastasis was relatively low in these patients with potentially resectable lung cancer.


Subject(s)
Adenocarcinoma/diagnosis , Adrenal Gland Neoplasms/diagnosis , Adrenal Glands/pathology , Endoscopic Ultrasound-Guided Fine Needle Aspiration , Lung Neoplasms/pathology , Small Cell Lung Carcinoma/diagnosis , Adenocarcinoma/secondary , Adrenal Gland Neoplasms/secondary , Adrenal Glands/diagnostic imaging , Adult , Aged , Aged, 80 and over , Endosonography , Female , Humans , Lung Neoplasms/surgery , Male , Middle Aged , Multimodal Imaging , Positron-Emission Tomography , Preoperative Care , Retrospective Studies , Small Cell Lung Carcinoma/secondary , Tomography, X-Ray Computed
2.
Endoscopy ; 45(5): 362-9, 2013.
Article in English | MEDLINE | ID: mdl-23616126

ABSTRACT

BACKGROUND AND STUDY AIMS: No prospective comparison of endoscopic ultrasonography-guided direct celiac ganglia neurolysis (EUS - CGN) vs. EUS-guided celiac plexus neurolysis (EUS - CPN) has been reported. The aim of the current study was to compare the effectiveness of EUS - CGN and EUS - CPN in providing pain relief from upper abdominal cancer pain in a multicenter randomized controlled trial. PATIENTS AND METHODS: Patients with upper abdominal cancer pain were randomly assigned to treatment using either EUS - CGN or EUS - CPN. Evaluation was performed at Day 7 postoperatively using a pain scale of 0 to 10. Patients for whom pain decreased to ≤ 3 were considered to have a positive response, and those experiencing a decrease in pain to ≤ 1 were considered to be completely responsive. Comparison between the two groups was performed using intention-to-treat analysis. The primary endpoint was the difference in treatment response rates between EUS - CGN and EUS - CPN at postoperative Day 7. Secondary endpoints included differences in complete response rates, pain scores, duration of pain relief, and incidence of adverse effects. RESULTS: A total of 34 patients were assigned to each group. Visualization of ganglia was possible in 30 cases (88 %) in the EUS - CGN group. The positive response rate was significantly higher in the EUS - CGN group (73.5 %) than in the EUS - CPN group (45.5 %; P = 0.026). The complete response rate was also significantly higher in the EUS - CGN group (50.0 %) than in the EUS - CPN group (18.2 %; P = 0.010). There was no difference in adverse events or duration of pain relief between the two groups. CONCLUSIONS: EUS - CGN is significantly superior to conventional EUS - CPN in cancer pain relief. CLINICAL TRIAL REGISTRATION: http://www.umin.ac.jp/ctr/index.htm (ID: UMIN-000002536).


Subject(s)
Abdominal Pain/therapy , Autonomic Nerve Block/methods , Celiac Plexus , Ganglia, Sympathetic , Pain Management/methods , Pancreatic Neoplasms/complications , Abdominal Pain/etiology , Aged , Aged, 80 and over , Anesthetics, Local , Bupivacaine , Endosonography , Ethanol/therapeutic use , Female , Humans , Intention to Treat Analysis , Male , Middle Aged , Ultrasonography, Interventional
3.
Endoscopy ; 45(8): 627-34, 2013 Aug.
Article in English | MEDLINE | ID: mdl-23807806

ABSTRACT

BACKGROUND AND STUDY AIMS: Only a few large cohort studies have evaluated the efficacy and safety of endoscopic necrosectomy for infected walled-off pancreatic necrosis (WOPN). Therefore, a multicenter, large cohort study was conducted to evaluate the efficacy and safety of endoscopic necrosectomy and to examine the procedural details and follow-up after successful endoscopic necrosectomy. PATIENTS AND METHODS: A retrospective review was conducted in 16 leading Japanese institutions for patients who underwent endoscopic necrosectomy for infected WOPN between August 2005 and July 2011. The follow-up data were also reviewed to determine the long-term outcomes of the procedures. RESULTS: Of 57 patients, 43 (75 %) experienced successful resolution after a median of 5 sessions of endoscopic necrosectomy and 21 days of treatment. Complications occurred in 19 patients (33 %) during the treatment period. Six patients died (11 %): two due to multiple organ failure and one patient each from air embolism, splenic aneurysm, hemorrhage from a Mallory - Weiss tear, and an unknown cause. Of 43 patients with successful endoscopic necrosectomy, recurrent cavity formation was observed in three patients during a median follow-up period of 27 months. CONCLUSIONS: Endoscopic necrosectomy can be an effective technique for infected WOPN and requires a relatively short treatment period. However, serious complications can arise, including death. Therefore, patients should be carefully selected, and knowledgeable, skilled, and experienced operators should perform the procedure. Further research into safer technologies is required in order to reduce the associated morbidity and mortality.


Subject(s)
Endoscopy, Digestive System , Pancreas/pathology , Pancreas/surgery , Pancreatic Diseases/surgery , Adult , Aged , Aged, 80 and over , Drainage , Endoscopy, Digestive System/adverse effects , Female , Humans , Japan , Male , Middle Aged , Necrosis/microbiology , Necrosis/surgery , Recurrence , Retrospective Studies , Stents , Therapeutic Irrigation , Young Adult
4.
Endoscopy ; 43(4): 369-72, 2011 Apr.
Article in English | MEDLINE | ID: mdl-21360425

ABSTRACT

Severe bleeding following endoscopic biliary sphincterotomy (EBS) can sometimes be difficult to manage, resulting in the need for an invasive intervention. The aim of this study was to retrospectively evaluate the feasibility and efficacy of endoscopic hemostasis using covered self-expandable metallic stents (SEMSs) for severe post- EBS bleeding. Eleven patients with bile duct stones underwent standard EBS using a standard sphincterotome-based technique at 4 endoscopic units of a university-affiliated hospital and a general hospital. Monotherapy or combined therapy were used to achieve hemostasis with either balloon tamponade, hypertonic saline epinephrine injection, or endoclip placement. When active bleeding could not be controlled, covered SEMSs were placed across the major papilla. Emergency endoscopy was performed on the day of admission or the subsequent day (ranging from 6 to 35 h after admission). Bleeding was classified as mild in 6 cases (54.5 %) and moderate in 5 (45.5 %). A covered SEMS 10mm in diameter and 6 cm long was placed across the papilla. After placement, complete hemostasis was achieved. The mean duration of stent placement was 8.2 days (range 5­10 days), and the SEMS was successfully removed in all cases. Although the present study has the limitations of a small sample size and lack of control patients, covered SEMS placement for endoscopic hemostasis may be useful in selected patients with uncontrolled post-EBS bleeding.


Subject(s)
Hemorrhage/therapy , Hemostasis, Endoscopic , Sphincterotomy, Endoscopic/adverse effects , Stents , Aged , Aged, 80 and over , Coated Materials, Biocompatible , Emergencies , Female , Hemorrhage/diagnosis , Hemorrhage/etiology , Humans , Male , Middle Aged
5.
Endoscopy ; 43(12): 1070-5, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21971925

ABSTRACT

BACKGROUND AND STUDY AIMS: Recent studies have evaluated the efficacy of peroral cholangioscopy (POCS) for diagnosis of biliary diseases. In order to obtain clear images with POCS, saline irrigation, which is performed to replace yellow bile, is carried out for an extended duration. The aim of this study was to evaluate the feasibility of replacing saline irrigation with CO2 insufflation during POCS. PATIENTS AND METHODS: A total of 36 patients who had bile duct lesions and were due to undergo POCS were enrolled in the study. Of these patients, 18 underwent POCS using saline irrigation followed by CO2 insufflation, and 18 patients underwent the reverse approach. The two methods were compared with regard to the time required to obtain a clear endoscopic image and the quality of the images. RESULTS: The median time required to obtain a clear endoscopic image using CO2 insufflation (5.0 min) was significantly shorter than that required for saline irrigation (22.5 min; P < 0.001). The quality of the endoscopic images obtained was similar in 27 cases. However, CO2 insufflation provided better images in four cases that showed an abundance of mucin or biliary sludge, and saline irrigation was superior to CO2 insufflation in five cases that showed severe stricture with bleeding and tall papillary lesions. CONCLUSIONS: CO2 insufflation during POCS can reduce procedure time and simplify cholangioscopy. The overall image quality was similar to that obtained with conventional saline irrigation.


Subject(s)
Bile Duct Diseases/diagnosis , Bile Ducts , Carbon Dioxide/administration & dosage , Endoscopy, Digestive System , Sodium Chloride/administration & dosage , Video Recording , Aged , Female , Humans , Insufflation , Male , Middle Aged , Therapeutic Irrigation
6.
Endoscopy ; 43(12): 1082-9, 2011 Dec.
Article in English | MEDLINE | ID: mdl-21971924

ABSTRACT

BACKGROUND: Recently, transesophageal endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) has been evaluated for mediastinal nodal staging (N staging) of lung cancer, as this technique is less invasive than mediastinoscopy and possibly more accurate than 18F-fluorodeoxyglucose positron emission tomography with computed tomography (PET-CT). However, EUS-FNA does not provide access to pretracheal and hilar lymph nodes. More recently, endobronchial ultrasound-guided transbronchial needle aspiration (EBUS-TBNA) has been introduced as a novel technique for accessing pretracheal and hilar lymph nodes. Although the combined endoscopic approach of EUS-FNA and EBUS-TBNA is presumably more accurate than PET-CT, only a few reports have quantitatively evaluated its diagnostic ability. Therefore, we prospectively assessed the diagnostic yield of this combined endoscopic approach for mediastinal N staging of lung cancer. METHODS: A consecutive series of 120 patients with suspected resectable lung cancer on CT findings underwent PET-CT and combined EUS-FNA/EBUS-TBNA. The accuracy and other diagnostic indices of the combined approach in mediastinal N staging were compared with those of PET-CT. RESULTS: Among the enrolled patients, a final pathological N stage was established in 110 patients. The accuracy of the combined approach using EUS-FNA and EBUS-TBNA was significantly higher than that of PET-CT (90.0 % vs. 73.6 %; P < 0.0001). The sensitivity, specificity, and positive and negative predictive values were respectively 71.8 %, 100 %, 100 %, and 86.6 % for the combined approach vs. 47.4 %, 87.5 %, 66.7 %, and 75.9 % for PET-CT. CONCLUSIONS: The combined endoscopic approach using EUS-FNA and EBUS-TBNA provided excellent diagnostic performance. Therefore, this approach is strongly recommended before surgery or mediastinoscopy to avoid futile thoracotomy and surgical intervention.


Subject(s)
Biopsy, Fine-Needle , Bronchoscopy , Endosonography , Lung Neoplasms/pathology , Lymph Nodes/pathology , Ultrasonography, Interventional , Adult , Aged , Aged, 80 and over , Biopsy, Fine-Needle/methods , Female , Humans , Lung Neoplasms/diagnostic imaging , Lymphatic Metastasis , Male , Mediastinum , Middle Aged , Multimodal Imaging , Neoplasm Staging , Positron-Emission Tomography , Sensitivity and Specificity , Tomography, X-Ray Computed
7.
Endoscopy ; 41(2): 179-82, 2009 Feb.
Article in English | MEDLINE | ID: mdl-19214901

ABSTRACT

Splenic tumors are occasionally found in clinical practice but the diagnosis is often difficult if only serologic and imaging tests are used. Therefore, pathologic sampling is required in such cases. Endoscopic ultrasonography (EUS) provides a good image of the spleen through the gastric wall, and a transgastric EUS-guided fine needle aspiration (EUS-FNA) biopsy may be easier than the percutaneous approach. Furthermore, a large-gauge needle may raise the capability of EUS-FNA for the histopathologic diagnosis. The aim of this study was to evaluate the yield of EUS-FNA using a large-gauge needle for a splenic tumor. Five patients with splenic tumor were subjected to EUS-FNA with a 19-gauge needle to obtain histopathologic materials. A pathologic sample was obtained in all cases, and the diagnoses were lymphoma (n = 2), sarcoidosis (n = 2), and inflammatory pseudotumor (n = 1). EUS-FNA using a 19-gauge needle is safe and useful for the diagnosis of splenic tumors.


Subject(s)
Biopsy, Fine-Needle/methods , Endosonography , Granuloma, Plasma Cell/pathology , Lymphoma/pathology , Sarcoidosis/pathology , Splenic Diseases/pathology , Aged , Female , Follow-Up Studies , Granuloma, Plasma Cell/diagnostic imaging , Humans , Lymphoma/diagnostic imaging , Male , Middle Aged , Needles , Pilot Projects , Predictive Value of Tests , Prospective Studies , Sarcoidosis/diagnostic imaging , Splenic Diseases/diagnostic imaging
8.
Endoscopy ; 40(5): 400-5, 2008 May.
Article in English | MEDLINE | ID: mdl-18494134

ABSTRACT

BACKGROUND AND STUDY AIM: Sarcoidosis is a systemic disorder of unknown cause that is characterized by a pathological hallmark, noncaseating granuloma. Bilateral hilar lymphadenopathy (BHL) is a major clinical feature, but it is sometimes difficult to exclude other diseases, especially in cases where there are no pulmonary abnormalities (stage I). Bronchoscopic transbronchial biopsy is currently a popular method by which to obtain pathological material, but its diagnostic power is insignificant. Endoscopic ultrasound-guided fine needle aspiration (EUS-FNA), also attempted recently, makes the sampling of pathological material easier and better, but the diagnoses are still based on cytological findings. Our study aimed to evaluate the yield of transesophageal EUS-FNA for histological confirmation of stage I sarcoidosis. METHODS: The study was a prospective comparative study to investigate the diagnostic sensitivities of FNA cytology and FNA histology. Subjects were consecutive patients with BHL without lung lesions on chest radiographs or chest CT who were referred to our hospitals between December 2003 and April 2006. Transesophageal EUS-FNA was performed with 19-gauge needles instead of the conventional 22-gauge needles. RESULTS: Forty-one patients were included in this study, and both histological and cytological materials were obtained successfully by EUS-FNA in all patients. Histopathological examination of the FNA sample showed noncaseating granuloma in 34 (94.4%) of the 36 patients with a final diagnosis of sarcoidosis. In contrast, only 28 of the 36 (77.8%) were diagnosed as having sarcoidosis on the basis of cytological findings. The difference was statistically significant (P = 0.0444). CONCLUSION: FNA histology is better suited than FNA cytology to establishing the diagnosis of stage I sarcoidosis, and EUS-FNA with a 19-gauge needle plays a important role in this process.


Subject(s)
Biopsy, Fine-Needle/methods , Endosonography , Sarcoidosis, Pulmonary/diagnostic imaging , Sarcoidosis, Pulmonary/pathology , Surgery, Computer-Assisted , Adult , Aged , Aged, 80 and over , Biopsy, Fine-Needle/instrumentation , Cohort Studies , Female , Humans , Male , Middle Aged , Needles , Predictive Value of Tests , Severity of Illness Index
9.
Aliment Pharmacol Ther ; 48(2): 138-151, 2018 07.
Article in English | MEDLINE | ID: mdl-29876948

ABSTRACT

BACKGROUND: Pre-operative tissue diagnosis for suspected malignant biliary strictures remains challenging. AIM: To develop evidence-based consensus statements on endoscopic tissue acquisition for biliary strictures. METHODS: The initial draft of statements was prepared following a systematic literature review. A committee of 20 experts from Asia-Pacific region then reviewed, discussed, and modified the statements. Two rounds of independent voting were conducted to reach a final version. Consensus was considered to be achieved when 80% or more of voting members voted "agree completely" or "agree with some reservation." RESULTS: Eleven statements achieved consensus. The choice of tissue sampling modalities for biliary strictures depends on the clinical setting, the location of lesion, and availability of expertise. Detailed radiological and endoscopic evaluation is useful to guide the selection of appropriate tissue acquisition technique. Standard intraductal biliary brushing and/or forceps biopsy is the first option when endoscopic biliary drainage is required with an overall (range) sensitivity and specificity of 45% (26%-72%) and 99% (98%-100%), and 48% (15%-100%) and 99% (97%-100%), respectively, in diagnosing malignant biliary strictures. Probe-based confocal laser endomicroscopy and fluorescence in situ hybridisation using 4 fluorescent-labelled probes targeting chromosomes 3, 7, 17 and 9p21 locus may be added to improve the diagnostic yield. Cholangioscopy-guided biopsy and EUS-guided tissue acquisition can be considered after prior negative conventional tissue sampling with an overall (range) sensitivity and specificity of 60% (38%-88%) and 98% (83%-100%), and 80% (46%-100%) and 97% (92%-100%), respectively, in diagnosing malignant biliary strictures. CONCLUSION: These consensus statements provide evidence-based recommendations for endoscopic tissue acquisition of biliary strictures.


Subject(s)
Cholangiography/standards , Cholestasis/pathology , Endoscopy, Gastrointestinal/standards , Practice Guidelines as Topic , Asia/epidemiology , Bile Duct Neoplasms/diagnosis , Bile Duct Neoplasms/pathology , Biopsy/methods , Biopsy/standards , Cholangiography/methods , Cholangiopancreatography, Endoscopic Retrograde/methods , Cholangiopancreatography, Endoscopic Retrograde/standards , Cholangiopancreatography, Endoscopic Retrograde/statistics & numerical data , Cholestasis/diagnosis , Consensus , Constriction, Pathologic/diagnosis , Constriction, Pathologic/pathology , Endoscopy, Gastrointestinal/methods , Humans , Image-Guided Biopsy/methods , Image-Guided Biopsy/standards , Pacific Islands/epidemiology , Sensitivity and Specificity
11.
Clin Cancer Res ; 3(5): 727-31, 1997 May.
Article in English | MEDLINE | ID: mdl-9815742

ABSTRACT

A goal of cancer chemoprevention is the deletion of latent premalignant or malignant clones before they expand to a clinically detectable tumor. However, such clonal deletion has not been demonstrated in clinical studies. We have evaluated serum levels of lectin-reactive alpha-fetoprotein (AFP-L3), which suggests the presence of latent hepatoma cells, in a randomized controlled trial that used acyclic retinoid to prevent second primary hepatomas in patients who had received treatments that cured initial hepatomas. The trial involved 21 patients in each acyclic retinoid (600 mg daily) and placebo group and consisted of a 12-month period of drug administration and a subsequent follow-up period. Serum AFP-L3 was determined at entry and at the end of the 12-month treatment period using lectin-affinity electrophoresis and antibody-affinity blotting. Although neither treatment affected serum levels of total AFP, acyclic retinoid significantly reduced AFP-L3 levels after a 12-month administration (P < 0.01). Acyclic retinoid not only deleted AFP-L3 from patients who had been positive for AFP-L3 at entry but also prevented the appearance of AFP-L3 in patients who had been negative at entry (P < 0.01). In contrast, placebo significantly raised the incidence of AFP-L3-positive patients after a 12-month administration from that at entry (P < 0.05). Patients positive for AFP-L3 after a 12-month treatment had a significantly higher risk of second primary hepatomas in the subsequent follow-up period (P = 0.03). Acyclic retinoid may have deleted a clone of latent hepatoma cells producing AFP-L3 and thereby inhibited second primary hepatomas. Serum AFP-L3 may be a useful intermediate biomarker in the chemoprevention of second primary hepatomas by acyclic retinoid.


Subject(s)
Antineoplastic Agents/therapeutic use , Carcinoma, Hepatocellular/prevention & control , Liver Neoplasms/prevention & control , Neoplasms, Second Primary/prevention & control , Tretinoin/analogs & derivatives , alpha-Fetoproteins/analysis , Carcinoma, Hepatocellular/blood , Carcinoma, Hepatocellular/drug therapy , Carcinoma, Hepatocellular/surgery , Disease-Free Survival , Female , Follow-Up Studies , Humans , Lectins , Liver Neoplasms/blood , Liver Neoplasms/drug therapy , Liver Neoplasms/surgery , Male , Middle Aged , Neoplasms, Second Primary/blood , Placebos , Time Factors , Tretinoin/therapeutic use
12.
FEBS Lett ; 294(3): 267-70, 1991 Dec 09.
Article in English | MEDLINE | ID: mdl-1836769

ABSTRACT

A small fraction (approximately 5%) of protein kinase C (PKC) in the adult rat brain synaptosomes is tightly associated with Triton X-100-insoluble components (most likely membrane-skeleton elements), and is solubilized only after denaturation with sodium dodecyl sulfate. The kinase domain of this PKC can be released as a soluble form after limited proteolysis with calpain, whereas the regulatory domain which binds phorbol ester remains insoluble. The PKC in this fraction was identified as the beta II-subspecies or its related molecule. Presumably, this enzyme subspecies is responsible for the phosphorylation of a major PKC substrate protein, growth-associated protein-43, which is located in nerve endings as well as in growth cones in association with the membrane-skeleton elements.


Subject(s)
Brain/enzymology , Protein Kinase C/metabolism , Synaptosomes/enzymology , Amino Acid Sequence , Animals , Calpain/metabolism , GAP-43 Protein , Immunoblotting , Male , Membrane Glycoproteins/metabolism , Molecular Sequence Data , Nerve Tissue Proteins/metabolism , Phosphorylation , Protein Kinase C/chemistry , Rats , Rats, Inbred Strains , Regulatory Sequences, Nucleic Acid , Solubility
13.
FEBS Lett ; 263(1): 31-4, 1990 Apr 09.
Article in English | MEDLINE | ID: mdl-2332050

ABSTRACT

Protein kinase C (PKC) from human promyelocytic leukemia HL-60 cells can be resolved into three fractions (peak, a, b and c) by hydroxyapatite column chromatography. Peak a and c enzymes are indistinguishable from the brain type II PKC having beta (beta I and beta II)-sequence and type III having alpha-sequence, respectively. Peak b enzyme is a previously unidentified PKC subspecies that has enzymological properties subtly different from type I (having gamma-sequence), type II and type III PKC. Upon treatment of HL-60 cells with 1 microM retinoic acid, this peak b enzyme is decreased dramatically within 24 h, whilst peak a enzyme (beta-PKC) is increased, and peak c (alpha-PKC) enzyme is slightly decreased within 48 h. The result implies that the PKC subspecies in HL-60 cells have distinct functions during cell differentiation.


Subject(s)
Cell Differentiation , Isoenzymes/metabolism , Protein Kinase C/metabolism , Tumor Cells, Cultured/cytology , Amino Acid Sequence , Antibodies , Cell Line , Enzyme Activation , Humans , Isoenzymes/genetics , Isoenzymes/isolation & purification , Kinetics , Leukemia, Promyelocytic, Acute , Molecular Sequence Data , Protein Kinase C/genetics , Protein Kinase C/isolation & purification , Tretinoin/pharmacology , Tumor Cells, Cultured/drug effects , Tumor Cells, Cultured/enzymology
14.
Biochem Pharmacol ; 44(2): 373-8, 1992 Jul 22.
Article in English | MEDLINE | ID: mdl-1322667

ABSTRACT

beta-Eudesmol, a major component of the crude drug "So-jutsu" (Atractylodis Lanceae Rhizoma), inhibited Na+, K(+)-ATPase activity most strongly among the various kinds of phosphatases examined. It also inhibited Ca(2+)-ATPase and H+, K(+)-ATPase, but to a lesser extent. Its effect on Mg(2+)-ATPase was minute. No effects on H(+)-ATPase or alkaline and acid phosphatase activities were observed. The effects of beta-eudesmol on horse kidney Na+, K(+)-ATPase were studied in detail, and the following results were obtained: (1) beta-eudesmol inhibited the Na+, K(+)-ATPase activity with an I50 value of 1.6 x 10(-4) M. The mode of its inhibition was uncompetitive with respect to ATP; (2) it prevented the stimulation of enzyme activity by Na+. The inhibition gradually increased in accord with the increase of Na+ concentration, and it was constant when Na+ was higher than 6.3 mM; (3) it did not alter the K+ concentration necessary for half-maximal activation (K0.5 for K+); and (4) it inhibited the enzyme activity with a mode of action different from ouabain. Phosphorylation of enzyme with [gamma-32P]ATP was inhibited by beta-eudesmol with an I50 of 1.4 x 10(-4) M. The inhibition was greater in 1 M NaCl than in 0.1 M NaCl. It had no effects on dephosphorylation steps, i.e. none of the non-specific, the ADP-sensitive (Na.E1-P----Na.E1) and the K(+)-dependent (E2-P----K.E2) dephosphorylation processes were affected. These results suggest that beta-eudesmol, a relatively specific inhibitor of Na+, K(+)-ATPase, interacts with the enzyme in the Na.E1 form and inhibits the reaction step Na.E1----Na.E1-P.


Subject(s)
Kidney/drug effects , Sesquiterpenes, Eudesmane , Sodium-Potassium-Exchanging ATPase/drug effects , Terpenes/pharmacology , Adenosine Triphosphatases/antagonists & inhibitors , Animals , Calcium-Transporting ATPases/antagonists & inhibitors , H(+)-K(+)-Exchanging ATPase , Horses , Kidney/enzymology , Phosphoric Monoester Hydrolases/antagonists & inhibitors , Potassium Chloride/pharmacology , Rats , Sodium Chloride/pharmacology , Stomach/drug effects , Terpenes/isolation & purification
15.
Biochem Pharmacol ; 53(4): 611-4, 1997 Feb 21.
Article in English | MEDLINE | ID: mdl-9105414

ABSTRACT

The inhibition of Na+,K(+)-ATPase activity by various constituents of Moutan Cortex and Paeoniae Radix was studied. 1,2,3,4,6-Penta-O-galloyl-beta-D-glucose (PGG), a major component of both crude drugs, strongly inhibited Na+,K(+)-ATPase activity (IC50 = 2.5 x 10(-6) M), whereas galloylpaeoniflorin, benzoic acid, and catechin were weakly inhibitory, and albiflorin, oxypaeoniflorin, paeoniflorin, paconol, and phenol were ineffective. The inhibition of Na+,K(+)-ATPase activity by PGG was decreased in the presence of BSA or phospholipids. The inhibition mode of PGG was noncompetitive with respect to ATP. The K0.5 value for Na+ was increased by the addition of PGG from 9.1 to 12.3 mM, whereas that for K+ was not altered. PGG also inhibited K(+)-dependent p-nitrophenyl phosphatase activity with an IC50 value of 5.3 x 10(-6) M, and the extent of the inhibition increased at higher concentrations of K+. The K0.5 value for K+ was decreased by the addition of PGG from 3.3 to 2.0 mM. These results suggested that the inhibition of Na+,K(+)-ATPase activity is caused by interaction of PGG with the enzyme in the E2 state. The inhibitory effect of Moutan Cortex or Paeoniae Radix is considered to be mainly attributable to PGG.


Subject(s)
Drugs, Chinese Herbal/pharmacology , Enzyme Inhibitors/pharmacology , Hydrolyzable Tannins , Sodium-Potassium-Exchanging ATPase/antagonists & inhibitors , Tannins/pharmacology , 4-Nitrophenylphosphatase/antagonists & inhibitors , Animals , Dose-Response Relationship, Drug , Horses
16.
J Biochem ; 130(3): 425-30, 2001 Sep.
Article in English | MEDLINE | ID: mdl-11530019

ABSTRACT

Previously we isolated and characterized a membrane-bound, arginine-specific serine protease from pig intestinal mucosa [J. Biol. Chem. 269, 32985-32991 (1994)]. For further characterization of this type of enzyme, we cloned a cDNA from rat intestinal mucosa encoding the precursor of a similar protease. The partial amino acid sequences determined for the pig enzyme were found to be shared almost completely by the rat enzyme. The serine protease domain of the rat enzyme, heterologously expressed in Escherichia coli, specifically cleaved Arg (or Lys)-X bonds with a marked preference for Arg-Arg or Arg-Lys, similar to the pig enzyme. The mRNA for the rat enzyme was shown to be distributed mainly in intestine, and the enzyme was detected in the duodenal mucosa as a 70 kDa protein. Immunohistochemical analysis of the small intestinal tissue showed that the enzyme is localized mainly on brushborder membranes.


Subject(s)
Arginine/metabolism , DNA, Complementary/isolation & purification , Intestinal Mucosa/enzymology , Serine Endopeptidases/genetics , Serine Endopeptidases/metabolism , Amino Acid Sequence , Animals , Cloning, Molecular , DNA, Complementary/genetics , DNA, Complementary/metabolism , Duodenum/enzymology , Enzyme Inhibitors/metabolism , Esophagus/enzymology , Humans , Male , Membrane Proteins , Mice , Molecular Sequence Data , Rats , Rats, Wistar , Sequence Homology , Species Specificity , Subcellular Fractions/enzymology , Substrate Specificity , Swine , Tissue Distribution , Trypsin/metabolism
17.
J Biochem ; 123(3): 546-54, 1998 Mar.
Article in English | MEDLINE | ID: mdl-9538241

ABSTRACT

As reported previously, caldecrin, a serum calcium-decreasing factor, from pancreas was found to be a serine protease, but the proteolytic activity was not necessary for its serum calcium decreasing activity. The caldecrin cDNA encoded a protease zymogen of the chymotrypsin/elastase superfamily consisting of a signal peptide, an activation peptide and a mature enzyme. On a homology search, we found that the sequence of rat caldecrin is almost identical to that of rat elastase IV (nucleotides: 99.3%175B amino acids: 90.3%) with the exception of the central region. However, it is not known whether or not elastase IV is transcribed and translated in vivo, and has proteolytic activity. In the present study, we constructed a rat elastase IV cDNA by means of combinatorial PCR, and compared the recombinant elastase IV with the recombinant caldecrin synthesized in a baculovirus expression system. The recombinant caldecrin protein was expressed in the cells and secreted mainly into the medium. In contrast, in the case of elastase IV, no recombinant protein was immunologically or enzymatically detected in the medium, while an immunoreactive protein with much lower protease activity was found in the cells in an amount comparable to that of the caldecrin protein. Using the RT-PCR method to discriminate caldecrin mRNA from elastase IV mRNA, we detected caldecrin mRNA expression in rat pancreas, but no elastase IV mRNA expression in any tissues examined. PCR analysis of rat genomic DNA revealed the presence of caldecrin and the absence of elastase IV sequences. These results indicate that caldecrin is expressed in the pancreas, but that elastase IV is an artifact produced during cloning. Furthermore, we investigated the protein-chemical and enzymological properties of the rat and human caldecrins using their recombinant proteins. Both recombinant proteins were secreted into the medium as proforms and showed protease activity after trypsin treatment. Some differences were found in the activation process and stability between human and rat caldecrins; human caldecrin was more easily activated by trypsin, but was much more labile than rat caldecrin. Although both caldecrins were found to be chymotrypsin-type proteases, on the basis of their substrate and inhibitor specificities, they were not inhibited by TPCK, suggesting that caldecrin is a novel type of serine protease.


Subject(s)
Pancreas/enzymology , Pancreatic Elastase/genetics , Pancreatic Elastase/metabolism , Serine Endopeptidases/genetics , Serine Endopeptidases/metabolism , Animals , Artifacts , Baculoviridae/genetics , Cloning, Molecular , Humans , Insecta , Polymerase Chain Reaction , Protease Inhibitors/pharmacology , RNA, Messenger/analysis , Rats , Recombinant Proteins/genetics , Recombinant Proteins/metabolism , Serine Endopeptidases/drug effects , Substrate Specificity , Tissue Distribution , Trypsin/metabolism
18.
Phytochemistry ; 37(2): 467-72, 1994 Sep.
Article in English | MEDLINE | ID: mdl-7765627

ABSTRACT

Investigation of the roots of Patrinia scabra afforded two new iridolactones, patriscabrol and isopatriscabrol. Their structures have been established by extensive analysis of their NMR spectra and an X-ray crystallographic analysis of patriscabrol. Four monoglucosides of each iridolactone were also isolated together with an apioside of glucosidic patriscabrol.


Subject(s)
Glycosides/chemistry , Lactones/chemistry , Plants, Medicinal/chemistry , Carbohydrate Sequence , Chromatography, High Pressure Liquid , Crystallography, X-Ray , Drugs, Chinese Herbal/chemistry , Glycosides/isolation & purification , Lactones/isolation & purification , Magnetic Resonance Spectroscopy , Molecular Sequence Data
19.
Phytochemistry ; 44(4): 735-8, 1997 Feb.
Article in English | MEDLINE | ID: mdl-9041720

ABSTRACT

Three alkaloids, neoharringtonine, homoneoharringtonine and 3'S-hydroxyneoharringtonine, were isolated from the leaves and stems of Cephalotaxus harringtonia var. drupacea. Their structures were established by spectroscopic methods, including two-dimensional NMR and CD spectra, and their antileukaemic activity was evaluated using P-388 leukaemia cells.


Subject(s)
Alkaloids/chemistry , Plants/chemistry , Alkaloids/isolation & purification , Animals , Chromatography, High Pressure Liquid , Circular Dichroism , Leukemia P388/pathology , Magnetic Resonance Spectroscopy , Molecular Conformation , Spectrometry, Mass, Fast Atom Bombardment , Tumor Cells, Cultured
20.
J Gastroenterol ; 32(5): 677-83, 1997 Oct.
Article in English | MEDLINE | ID: mdl-9349997

ABSTRACT

A 17-year-old woman was admitted because of a liver tumor found incidentally by ultrasonography. Liver function was normal and there were no markers of hepatitis viruses or malignancy. Abdominal ultrasonography, computed tomography (CT), and magnetic resonance imaging revealed a mass (2 cm in diameter) in the lateral segment of the left lobe of the liver. The lesion was not detected by hepatic arteriography. However, dynamic CT with fast scanning and dynamic CO2-enhanced ultrasonography demonstrated initial central enhancement of the mass followed by centrifugal spread of enhancement to the periphery. Color Doppler flow imaging detected a central color spot, shown to be an artery by a pulsed Doppler spectrum analysis. Fine-needle biopsy confirmed a diagnosis of focal nodular hyperplasia. Dynamic CT with fast scanning, dynamic CO2-enhanced ultrasonography, and color Doppler flow imaging were useful in detecting the vascular pattern specific to focal nodular hyperplasia. Investigation of further cases with these novel imaging modalities should help to establish a comprehensive diagnostic procedure and thus avoid unnecessary surgery for focal nodular hyperplasia, which is a completely benign lesion.


Subject(s)
Liver Diseases/diagnosis , Liver , Adolescent , Biopsy, Needle , Diagnosis, Differential , Female , Humans , Hyperplasia/diagnosis , Liver/blood supply , Liver/diagnostic imaging , Liver/pathology , Liver Neoplasms/blood supply , Liver Neoplasms/diagnosis , Magnetic Resonance Imaging , Tomography, X-Ray Computed , Ultrasonography, Doppler, Color
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