Distribution of (125)I-labeled crotamine in mice tissues.

Crotamine is a strong basic polypeptide from Crotalus durissus terrificus (Cdt) venom composed of 42 amino acid residues tightly bound by three disulfide bonds. It causes skeletal muscle spasms leading to spastic paralysis of hind limbs in mice. The objective of this paper was to study the distribution of crotamine injected intraperitoneally (ip) in mice. Crotamine was purified from Cdt venom by gel filtration followed by ion exchange chromatography, using a fast-performance liquid chromatography (FPLC) system. Purified crotamine was irradiated at 2 kGy in order to detoxify. Both native and irradiated proteins were labeled with (125)I using chloramine T method, and separated by gel filtration. Male Swiss mice were injected ip with 0.1 mL (2 x 10(6)cpm/mouse) of (125)I native or irradiated crotamine. At various time intervals, the animals were killed by ether inhalation and blood, spleen, liver, kidneys, brain, lungs, heart, and skeletal muscle were collected in order to determine the radioactivity content. The highest levels of radioactivity were found in the kidneys and the liver, and the lowest in the brain.

New view on crotamine, a small basic polypeptide myotoxin from South American rattlesnake venom.

Crotamine is a toxin from the Crotalus durissus terrificus venom, composed of 42 amino acid residues and three disulfide bridges. It belongs to a toxin family previously called Small Basic Polypeptide Myotoxins (SBPM) whose members are widely distributed through the Crotalus snake venoms. Comparison of SBPM amino acid sequences shows high similarities. Crotamine induces skeletal muscle spasms, leading to spastic paralysis of the hind limbs of mice, by interacting with sodium channels on muscle cells. The crotamine gene with 1.8 kbp is organized into three exons, which are separated by a long phase-1 and short phase-2 introns and mapped to chromosome 2. The three-dimensional structure of crotamine was recently solved and shares a structural topology with other three disulfide bond-containing peptide similar to human beta-defensins and scorpion Na+ channel toxin. Novel biological activities have been reported, such as the capacity to penetrate undifferentiated cells, to localize in the nucleus, and to serve as a marker of actively proliferating living cells.

Aggressive non-Hodgkin's lymphoma after successful eradication of Helicobacter pylori and regression of gastric lymphoma of mucosa-associated lymphoid tissue.

A 57-year-old woman presented to our clinic with low-grade gastric lymphoma of mucosa-associated lymphoid tissue (stage IE) and Helicobacter pylori infection. She received a 2-week course of omeprazole and clarithromycin, resulting in eradication of H. pylori and histological disappearance of the lymphoma. However, 9 months later (May 1996), multiple mass lesions were found around the pancreas and hepato-duodenal ligament on abdominal computed tomography. Inguinal lymph node biopsy revealed aggressive nodal type B-cell non-Hodgkin's lymphoma, diffuse large cell type. She received chemotherapy with cyclophosphamide, adriamycin, vincristine, and prednisolone, but failed to achieve remission and died in December 1996. There was no evidence of recurrent gastric lymphoma. This case emphasizes the importance of performing follow-up examinations to detect other neoplasms in patients with gastric lymphoma of mucosa-associated lymphoid tissue.

Effects of (60)Co gamma radiation on crotamine.

Ionizing radiation can change the molecular structure and affect the biological properties of biomolecules. This has been employed to attenuate animal toxins. Crotamine is a strongly basic polypeptide (pI 10.3) from Crotalus durissus terrificus venom composed of 42 amino acid residues. It induces skeletal muscle spasms leading to a spastic paralysis of hind limbs in mice. The objective of the present study was to carry out a biochemical study and a toxic activity assay on native and irradiated crotamine. Crotamine was purified from C.d. terrificus venom by Sephadex G-100 gel filtration followed by ion-exchange chromatography, and irradiated at 2 mg/ml in 0.15 M NaCl with 2.0 kGy gamma radiation emitted by a 60Co source. The native and irradiated toxins were evaluated in terms of structure and toxic activity (LD50). Irradiation did not change the protein concentration, the electrophoretic profile or the primary structure of the protein although differences were shown by spectroscopic techniques. Gamma radiation reduced crotamine toxicity by 48.3%, but did not eliminate it.

A single-step purification of bothropstoxin-1.

Bothrops venoms are complex mixtures of components with a wide range of biological activities. Among these substances, myotoxins have been investigated by several groups. Bothropstoxin-1 (Bthtx-1) is a phospholipase A2-like basic myotoxin from Bothrops jararacussu. The purification of this component involves two chromatographic steps. Although providing a pure material, the association of these two steps is time consuming and a single-step method using high performance chromatography media would be useful. In the present study, we describe a single-step purification method for Bthtx-1. Bothrops jararacussu venom was dissolved in 1 ml buffer. After centrifugation, the supernatant was injected into a Resource-S cation exchange column connected to an FPLC system and eluted with a linear salt gradient. The complete procedure took 20 min, representing a considerable time gain when compared to a previously described method (Homsi-Brandenburgo MI et al. (1988) Toxicon, 26: 615-627). Bthtx-1 purity and identity, assessed by SDS-PAGE and N-terminal sequencing, resulted in a single band with a molecular mass of about 14 kDa and the expected sequence of the first 5 residues, S-L-F-E-L. Although the amount of protein purified after each run is lower than in the previously described method, we believe that this method may be useful for small-scale purifications.

Heterotopic intestinal membrane in a retroperitoneal tumor.

Plain abdominal X-rays of a 13-year-old girl with chief complaints of back pain revealed calcification in the upper left abdomen. A calcified tumor was confirmed at the dorsal side of the pancreatic tail upon admission. A completely formed colic membrane free of all other germ layers was discovered within the tumor, leading to a diagnosis of heterotopic colonic membrane. To our knowledge, there have been no other cases of heterotopic intestinal tissue of this type, so we consider this an extremely rare case worth reporting.

[Study of patency of minor duodenal papilla--comparison between pigment method combined with pH sensor and pancreatographic method].

We investigated the patency of minor duodenal papilla in 25 cases that had presented normal pancreatogram by using the endoscopic retrograde pancreatographic (ERP) method and a pH sensor-combined indigo carmine pigment method. The pigment method allowed us to classify the function of minor papilla into three types according to pancreatic juice excretion pre- and post-secretin administration (Type I), excretion after secretin administration (Type II), and no excretion even after secretin administration (Type III). Twelve cases belonged to Type I, 4 cases to Type II and 9 cases to Type III. By ERP method, terminal shapes of the Santorini's duct were classified as stick type, cystic type and branch type. 19 cases were considered stick type, 2 cases cystic type and 4 cases branch type. Eighteen cases were identified as patent minor papilla under ERP method. There were two cases alleged to be patent under the ERP but without pancreatic juice excretion. Therefore, the necessity of pigment method was confirmed for investigation of the function of minor papilla.

[Examination of cholangiogram of obstructive jaundice using MRI--compared with PTC and ERCP].

PTC and ERCP are most often used in diagnosis of obstructive jaundice. We studied the possibility of clinical diagnosis using MRI in 33 cases of obstructive jaundice. A clinical diagnosis of malignant tumors could be given in 17 cases out of 20 (85%) using MRI if respiratory standstill was possible. An MRI cholangiogram was particularly effective in describing tearful parting bile ducts and was clearer than PTC in describing negative gallbladders. Choledochal stones could be diagnosed in 58% of cases, which was less than the rate for malignant tumors. MRI is not an invasive examination, can be used in diagnosis of obstructive jaundice, and helps in selecting treatment methods such as PTCD and ERBD.

[The utility of endoscopic ultrasonography (EUS) in endoscopic mucosal resection of early gastric cancer].

The utility of EUS was evaluated in 27 patients with early gastric cancer undergoing endoscopic mucosal resection over the past 2 years. Accuracy of the assessment of depth of cancerous invasion was studied in 16 patients undergoing EUS before endoscopic mucosal resection. Patients showing no changes in the submucosal (sm) layer or below on EUS included 15 with mucosal (m) cancer and one with sm cancer showing very slight infiltration. Seven patients with m cancer, a negative stump, and no ulcer in the cancer focus at endoscopic mucosal resection, were followed up for more than 1 year after endoscopic mucosal resection. On EUS, four patients showed Ul-IIs changes resembling benign ulcers, two showed Ul-IIIs changes and two showed no changes in the sm layer or below. All patients were negative for cancer in follow up biopsies. No lymphadenopathy was observed. EUS was effective in diagnosing the depth of cancerous invasion in patients undergoing endoscopic mucosal resection and also in clarifying changes in the sm and deeper layers during follow up.

[Imaging diagnosis of the insulinoma--the clinical usefulness of intraductal ultrasonography of the pancreas and color Doppler sonography].

5 cases of small-sized insulinoma of the pancreas were studied to evaluate the ability of diagnostic imaging techniques to determine tumor location. Techniques used were extracorporeal ultrasonography, endoscopic ultrasonography (EUS), angiography, percutaneous transhepatic portal catheterization and CT. EUS was superior to the other modalities in diagnosing the location of insulinomas. We employed Color Doppler (CD) ultrasonography in one case and obtained color expression in the tumor and a pulsatile wave in the FFT analysis. Intraductal sonography of the pancreas (IDSP) was also performed in this case resulting peripheral echo-lucent, central echogenic mass. From these results, we expect that CD and IDSP will be useful in clinical application for diagnosing various tumors.

[Examination of endoscopical ultrasonography (EUS) on carcinoid tumors of gastrointestinal tract].

In order to clarify the usefulness of EUS in the diagnosis of gastrointestinal carcinoid tumor, we examined 15 patients, who had carcinoid tumors of gastrointestinal tract (stomach: 5, duodenum: 2, rectum: 8), on the diagnosis in quality and depth by comparing endoscopical ultrasonography (EUS) with resected specimen. Carcinoid tumors of gastrointestinal tract were detected as homogenous hypoechoic tumors with sharp border in all site by EUS. Especially, it was characteristic that 14 of the 17 lesions (82%) which invaded into submucosa were mainly located in the third layer, and the second layer covered the tumor at the foot, and near the top, it touched the tumor and got indistinct. We decided the depth of invasion by comparing the hypoechoic tumor with normal structure of 5 layers. The accuracy rate was 88%. In conclusion, EUS was thought to be useful in the diagnosis and choice for treatment of carcinoid tumors of gastrointestinal tract.

[Value of MR cholangiography in the diagnosis of pancreatic diseases compared with endoscopic retrograde cholangiopancreatography].

PURPOSE: To compare magnetic resonance cholangiography (MRCP) with endoscopic retrograde cholangiography (ERCP) in the diagnosis of pancreatic diseases. METHODS: The authors retrospectively reviewed MRCP and ERCP images obtained in 56 patients with pancreatic diseases and 192 controls. Detectability of the pancreatic duct and its branches and diagnostic usefullness were evaluated. RESULTS: MRCP was successful in almost all the patients without ascitic fluid or ileus. Findings of the main pancreatic ducts (i.e., dilation, stenosis and obstruction) were similarly demonstrated by ERCP and MRCP. The degree of ductal narrowing tends to be overestimated with MRCP. CONCLUSION: MRCP appears to be less invasive than ERCP. MRCP could provide sufficient information and play a complementary role in the diagnosis of pancreatic diseases.