Proteomic profiling of k-11706 responsive proteins.

Erythropoietin promotes the production of red blood cells. Recombinant human erythropoietin is illicitly used to improve performance in endurance sports. Expression of the ERYTHROPOIETIN gene is negatively controlled by the transcription factor GATA-binding protein (GATA). Specific GATA inhibitors have recently been developed as novel drugs for the management of anemia. These drugs could, therefore, be illicitly used like recombinant human erythropoietin to improve performance in sports. To examine alterations in levels of plasma protein after administration of GATA inhibitors, proteomic analyses were conducted on mouse plasma samples treated with the potent GATA inhibitor K-11706. The analysis based on gel electrophoresis identified 41 protein spots differentially expressed when compared with normal plasma. Each spot was identified with liquid chromatography coupled to tandem mass spectrometry and 2 of them, fetuin-B and prothrombin, were verified by Western blotting. The results showed that the expression of fetuin-B in mice plasma was increased by K-11706, but not by recombinant human erythropoietin or hypoxia. These results suggest the potential of proteomic-based approaches as tools to identify biomarkers for the illegal use of novel drugs (e.g., GATA inhibitors). Also, fetuin-B could be a sensitive marker for the detection of abuse of GATA inhibitors.

Caffeine and taurine enhance endurance performance.

Caffeine enhances endurance performance; however, its effect on accumulated lactate remains unclear. Conversely, taurine, which also enhances endurance performance, decreases accumulated lactate. In this study, the effect of combination of caffeine and taurine on endurance performance was assessed. Mice ran on a treadmill, and the accumulated lactate was measured. In addition, muscle fibers from the gastrocnemius muscle of the mice were stained with ATPase and analyzed. The use of caffeine and taurine over a 2 week period enhanced endurance performance. Moreover, taurine significantly decreased the accumulated concentration of lactate over long running distances. However, the diameter of the cross-sections and ratios of Types I, IIA, and IIB muscle fibers were not affected.

Hydrolysis rate constants at 10-25 °C can be more than doubled by a short anaerobic pre-hydrolysis at 35 °C.

Hydrolysis is the first step of the anaerobic digestion of complex wastewater and considered as the rate limiting step especially at low temperature. Low temperature (10-25 °C) hydrolysis was investigated with and without application of a short pre-hydrolysis at 35 °C. Batch experiments were executed using cellulose and tributyrin as model substrates for carbohydrates and lipids. The results showed that the low temperature anaerobic hydrolysis rate constants increased by a factor of 1.5-10, when the short anaerobic pre-hydrolysis at 35 °C was applied. After the pre-hydrolysis phase at 35 °C and decreasing the temperature, no lag phase was observed in any case. Without the pre-hydrolysis, the lag phase for cellulose hydrolysis at 35-10 °C was 4-30 days. Tributyrin hydrolysis showed no lag phase at any temperature. The hydrolysis efficiency of cellulose increased from 40 to 62%, and from 9.6 to 40% after 9.1 days at 15 and 10 °C, respectively, when the pre-hydrolysis at 35 °C was applied. The hydrolysis efficiency of tributyrin at low temperatures with the pre-hydrolysis at 35 °C was similar to those without the pre-hydrolysis. The hydrolytic activity of the supernatant collected from the digestate after batch digestion of cellulose and tributyrin at 35 °C was higher than that of the supernatants collected from the low temperature (≤25 °C) digestates.

Characterization of two forms of hemagglutinin/protease produced by Vibrio cholerae non-O1.

Two forms (34 kDa and 32 kDa) of hemagglutinin/protease produced by Vibrio cholerae non-O1 were characterized. The hemagglutinin/protease purified by immunoaffinity column chromatography using a monoclonal antibody was essentially a 34-kDa form. By incubation of the purified 34-kDa form at 37 degrees C, it was processed (autodigested) to the 32-kDa form. The N-terminal 20 amino acid sequences of both the 34- and 32-kDa forms were identical, suggesting that proteolytic processing at the C-terminal region of the 34-kDa hemagglutinin/protease resulted in the 32-kDa form. With this shift, protease activity increased, but hemagglutinating activity decreased, suggesting that the C-terminal region of the hemagglutinin/protease is related to hemagglutinating activity.

Production of monoclonal antibodies against a hemagglutinin/protease of Vibrio cholerae non-01.

Two hybridoma cell lines producing monoclonal antibodies (MAbs) against a hemagglutinin/protease (HA/P) from Vibrio cholerae non-01 were produced and characterized. The two MAbs contained the kappa light chain and were IgG1 type. They similarly neutralized HA/P protease activity derived from both V. cholerae non-01 and V. cholerae 01, whereas they were unable to neutralize the hemagglutinating activity of HA/P, suggesting that the epitopes for protease and hemagglutination activities are different. Western blotting analysis and the cross-neutralization test with the two MAbs confirmed the identity of HA/P produced by V. cholerae non-01 and 01. This study also suggests that HA/P of V. cholerae and a protease of V. parahaemolyticus are immunologically unrelated.

Vibrio cholerae O139 produces a protease which is indistinguishable from the haemagglutinin/protease of Vibrio cholerae O1 and non-O1.

Haemaglutinin/protease (HA/P) is one of the virulence factors of Vibrio cholerae O1 and pathogenic strains of V. cholerae non-O1. In this study, we examined protease activity of a new serogroup of Vibrio cholerae recently designated as O139 synonym Bengal. The protease activity was produced by all eight isolates of V. cholerae O139 from Bangladeshi patients. Purification and partial characterization of the protease from V. cholerae O139 demonstrated the purified protease (O139-P) was indistinguishable from that previously reported for HA/P of V. cholerae non-O1 (NAG-HA/P) and V. cholerae O1 (Vc-HA/P). These results prove that V. cholerae O139 produces a protease belonging to HA/P, and suggest that the protease is another virulence factor found in newly emerged V. cholerae O139, as in V. cholerae O1.

Nicking sites in a subunit of cholera toxin and Escherichia coli heat-labile enterotoxin for Vibrio cholerae hemagglutinin/protease.

We analyzed the nicking site of the A subunit of Escherichia coli heat-labile enterotoxin for hemagglutinin/protease produced by Vibrio cholerae non-O1 (NAG-HA/P). The determined nicking site was the Thr193-Ile194 junction, which was distinct from that for a protease of V. cholerae (Ichinose et al., European Journal of Epidemiology 8, 743-747, 1992). We further analyzed proteolytic cleavage by NAG-HA/P of a synthetic peptide corresponding to the nicking region of cholera toxin A subunit and determined the cleavage site to be preferentially between Ser194 and Met195, and in addition between Ser193 and Ser194.

[Pitfall in 201Tl-99mTc subtraction scintigraphy: a case of 99mTc-pertechnetate uptake in a parathyroid hyperplasia].

201Tl-99mTc subtraction scintigraphy has been recognized as a useful procedure in the preoperative localization of hyperfunctioning parathyroid glands. We experienced a case which showed 99mTc-pertechnetate uptake in a parathyroid hyperplasia. This case warned us to focus a lot of attention on the detection for preoperative localization. There has been no such case reported in the previous Japanese literatures. Hypervascularity and thick fibrous capsule presumed explanation for a rare case of marked pertechnetate uptake into a parathyroid hyperplasia.

[Inflammatory pseudotumor of the ureter: a case report].

A 57-year-old male was admitted because of the right flank pain. The image examinations, retrogradeurography, abdominal CT and MRI, showed a mass located at the upper right ureter. Although the tumor was not typical as ureteral cancer, we could not make a diagnosis of a benign tumor by image examinations. Therefore nephroureterectomy that was surgical method for ureteral cancer was performed. The tumor was diagnosed as inflammatory pseudotumor of the ureter by histological findings. Inflammatory pseudotumor is extremely rare for ulogeital organs. And this lesion is difficult to distinguish from malignancy only by image examinations. Therefore, the surgical resection and pathological studies are necessary.

[Chronic hepatitis--clinicopathological studies of 687 cases (author's transl)].

Chronic hepatitis is one of liver diseases with arguments from the clinical and histopathological aspects. Histopathological examinations were made on 687 biopsy cases clinically diagnosed as chronic hepatitis. Histopathological classification was based on our own criteria by referring to discussions in the series of Inuyama symposia on hepattis and others. The correlation between histological diagnosis and clinical data was also examined. Histopathological diagnoses made of the 687 cases were classified as follows; normal liver or liver with no pathognomonic changes of 77 cases (11.2%), non-specific reactive hepatitis of 56 cases (8.0%), viral hepatitis of 488 cases (71.0%), alcoholic hepatitis of 25 cases (3.6%), fatty liver of 23 cases (3.3%), massive liver necrosis of 3 cases, liver fibrosis of 2 cases, congestive liver of 1 case, and unclassified 12 cases due to inadequate specimens or other reasons. Among 488 viral hepatitis cases, histological stages were as follows; acute hepatitis (38 cases, 7.8%), persistent hepatitis (23 cases, 4.7%), chronic inactive hepatitis (142 cases, 29.1%), chronic active hepatitis (165 cases, 33.8%), chronic hepatitis with subloblar necrosis (33 cases, 6.8%), precirrhosis (51 cases, 10.5%), cirrhosis (27 cases, 5.5%). The relationship between histological aspects and clinical features was discussed by sex, age, and others. Of 41 follow up cases, significant values of histological type, presence of HB ag., or alcoholic were discussed as for the causative factors evolving liver cirrhosis.

[Clinicopathological study on 100 Japanese patients with peritoneal mesothelioma in Japan].

We report a clinicopathological study on 100 japanese patients with peritoneal mesothelioma encountered between 1911 and 1982. Fifty-six were males and 44 were females, they ranged in age from 10 months to 83 years. As the inducement materials, asbestos was reported in 5 and thorium in 2 patients. Clinical symptoms were abdominal distension in 56 and abdominal pain in 50 patients. Thrombocytosis and hypoglycemia were observed in 7 and 3 patients, respectively. Macroscopically, the peritoneal mesothelioma was localized in 7, diffuse in 86 and not described in 7 patients. The pathological classification was benign in 5 and malignant in 77 cases. According to Stout's classification, the peritoneal mesothelioma was tubular in 43 mixed in 25 and fibrous type in 11 cases. Based on our findings we suggest that peritoneal mesotheliomas are on the increase in Japan.

A simple purification method of Vibrio cholerae non-O1 hemagglutinin/protease by immunoaffinity column chromatography using a monoclonal antibody.

A new simple purification method (I) for Vibrio cholerae non-O1 hemagglutinin/protease (NAG-HA/P) was developed. The method (I) requires only an immunoaffinity column chromatography using a monoclonal antibody against NAG-HA/P. The method (I) is much simpler than previously reported purification method (II) (Honda, T. et al, Infection and Immunity 57: 2799-2803, 1989) which required four or more complicated chromatographic procedures. Method (I) also gave an improved recovery rate (about 27%) compared with (II). The molecular weight of NAG-HA/P purified by method (I) was mainly 34 kilodaltons (kDa) with a little of 32 kDa, whereas that of NAG-HA/P purified by (II) was usually 32 kDa. Immunological analysis by the Ouchterlony double gel diffusion test and Western blotting test using polyclonal antibody against 32 kDa protein revealed that the 34 and 32 kDa proteins are immunologically indistinguishable and thus it is supposed that 34 K protein is an isoform or a preform of the 32 K protein.

Ganglioneuroblastoma associated with malignant mesenchymoma.

A case of ganglioneuroblastoma associated with malignant mesenchymoma is reported. The tumor originated from the retroperitoneum and was comprised not only of neuroblastoma and ganglioneuroma, but also of typical rhabdomyosarcoma, liposarcoma, undifferentiated mesenchymoma, as well as cartilaginous tissue. No distinct border existed between thses different components. The histogenesis of the tumor is discussed. We suggest that the tumor derived from a remnant of neural crest (ectomesenchyme).

Culture of isolated glomeruli from normal and nephritic rabbits. I. Characterization of outgrowing cells.

The existence of four types of cells in the glomerular outgrowths from both normal and nephritic rabbits was confirmed, and their characteristics by phase contrast, scanning electron, and transmission electron microscopy are described. Type I (arborized), Type II (fusiform), Type III (round), and Type IV (paving stone-like) cells presumably originate from visceral epithelial cells, mesangial cells, monocytes, and parietal epithelial cells of Bowman's capsule, respectively, though conclusive evidence is still lacking. The Type III cells, unlike the other types of cells, have a remarkable phagocytic capacity as well as Fc and C3 receptors.

Inflammatory fibroid polyp of the stomach.

Fourteen cases of inflammatory fibroid polyp of the stomach were studied in terms of immunohistochemistry and ultrastructure. They occurred as polypoid lesions in the antrum, except for two found in the body of the stomach. Out of the 14 cases, two were found to be multiple and the remainder solitary. In all but one lesion, the mucosal layer was involved and six lesions were entirely localized within the mucosal layer, suggesting that inflammatory fibroid polyps of the stomach originate in the mucosal layer. Neither S100 protein, factor VIII-related antigen, alpha 1-antitrypsin nor lysozymes were found in the cytoplasm of the proliferating cells. The ultrastructures of the proliferating cells were fibroblastic rather than neurogenic, angiogenic, or myofibroblastic. These findings suggest that the cells are fibroblasts, though the possibility that they are facultative fibroblasts remains. An interesting observation made with electron microscope was the infection of micro-organisms similar to mycoplasma. This seems to deserve further investigation as a possible etiologic factor of the disease.