ABSTRACT
We recently reported a Philyra pisum lectin (PPL) that exerts mitogenic effects on human lymphocytes, and its molecular characterization. The present study provides a more detailed characterization of PPL based on the results from a monosaccharide analysis indicating that PPL is a glycoprotein, and circular dichroism spectra revealing its estimated alpha-helix, beta-sheet, beta-turn, and random coil contents to be 14.0%, 39.6%, 15.8%, and 30.6%, respectively. These contents are quite similar to those of deglycosylated PPL, indicating that glycans do not affect its intact structure. The binding properties to different pathogen-associated molecular patterns were investigated with hemagglutination inhibition assays using lipoteichoic acid from Gram-positive bacteria, lipopolysaccharide from Gram-negative bacteria, and both mannan and beta-1,3-glucan from fungi. PPL binds to lipoteichoic acids and mannan, but not to lipopolysaccharides or beta-1,3-glucan. PPL exerted no significant antiproliferative effects against human breast or bladder cancer cells. These results indicate that PPL is a glycoprotein with a lipoteichoic acid or mannan-binding specificity and which contains low and high proportions of alpha-helix and beta-structures, respectively. These properties are inherent to the innate immune system of P. pisum and indicate that PPL could be involved in signal transmission into Gram-positive bacteria or fungi.
Subject(s)
Humans , beta-Glucans , Breast , Circular Dichroism , Fungi , Glycoproteins , Gram-Negative Bacteria , Gram-Positive Bacteria , Hemagglutination , Immune System , Lipopolysaccharides , Lymphocytes , Mannans , Polysaccharides , Sensitivity and Specificity , Teichoic Acids , Urinary Bladder NeoplasmsABSTRACT
A lectin from the hemolymph of purse crab, Philyra pisum, was found to have anti-proliferative activity on human lung cancer cells by our laboratory. In this study, P. pisum lectin (PPL) was molecularly characterized including molecular mass, amino acid sequences, amino acid composition, and the effects of metal ions, temperature, and pH on the activity. We found that PPL showed mitogenic activity on human lymphocytes and BALB/c mouse splenocytes. The mitogenic activity (maximum stimulation index, SI=9.57+/-0.59) of PPL on human lymphocytes was higher than that of a standard well-known plant mitogen, concanavalin A (maximum SI=8.80+/-0.59). The mitogenic activity mediated by PPL is required for optimum dosing, and higher or lower concentrations caused decreases in mitogenic response. PPL also induced mitogenic activity on mouse splenocytes, however, the maximum SI (1.77+/-0.09) on mouse splenocytes of PPL was lower than that (2.14+/-0.15) of concanavalin A. In conclusion, PPL is a metal ion-dependent monomer lectin with mitogenic activity, and could be used as a lymphocyte or splenocyte stimulator.
Subject(s)
Animals , Humans , Mice , Amino Acid Sequence , Concanavalin A , Hemolymph , Hydrogen-Ion Concentration , Ions , Lung Neoplasms , Lymphocytes , PlantsABSTRACT
The mushroom Cordyceps militaris has been used for a long time in eastern Asia as a nutraceutical and in traditional Chinese medicine as a treatment for cancer patients. In the present study, a cytotoxic antifungal protease was purified from the dried fruiting bodies of C. militaris using anion-exchange chromatography on a DEAE-Sepharose column. Electrophoretic analyses indicated that this protein, designated C. militaris protein (CMP), has a molecular mass of 12 kDa and a pI of 5.1. The optimum conditions for protease activity were a temperature of 37degrees C and pH of 7.0~9.0. The enzyme activity was specifically inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride. Amino acid composition of intact CMP and amino acid sequences of three major peptides from a tryptic digest of CMP were determined. CMP exerted strong antifungal effect against the growth of the fungus Fusarium oxysporum, and exhibited cytotoxicity against human breast and bladder cancer cells. These results indicate that C. militaris represents a source of a novel protein that might be applied in diverse biological and medicinal applications.
Subject(s)
Humans , Agaricales , Amino Acid Sequence , Breast , Chromatography , Cordyceps , Dietary Supplements , Asia, Eastern , Fruit , Fungi , Fusarium , Hydrogen-Ion Concentration , Medicine, Chinese Traditional , Peptides , Phenylmethylsulfonyl Fluoride , Serine Proteases , Urinary Bladder NeoplasmsABSTRACT
Multiple-antibiotic-resistance (Mar) mutants were prepared from the wild-type Escherichia coli K-12 MC4100 or W3110, were analyzed for the antibiotic resistance by minimal inhibitory concentration (MIC), and compared with their isogenic parent strains and E. coli clinical isolates. Mutation frequency was approximately 10-7 to 10-9 MIC levels of chloramphenicol (Cm), ampicillin (Ap), tetracycline (Tc), or ciprofloxacin (Cp) antibiotics against E. coli Mar mutants were 50 ug/ml, 300 ug/ml, 200 ug/ml, or 30 ug/ml, respectively, at their highest MIC levels. Cross-resistances of each Mar mutants against Cm, Ap, Tc, Cp, erythromycin (Em), nalidixic acid (Na), and rifampin (Rf) were evaluated. Almost all of Mar mutants showed approximately 9 to 140-fold increase of MIC with contrast to the MIC of isogenic parent strains and E. coli clinical isolates, except Rf antibiotics. The induction multiplicity of Mar mutants by salicylate (SAL) was approximately the same, 2 to 10-fold, and 2 to 25-fold increase of MIC, in case of E. coli clinical isolates, Mar mutants derived from MC4100, and W3110, respectively.
Subject(s)
Humans , Ampicillin , Anti-Bacterial Agents , Chloramphenicol , Ciprofloxacin , Drug Resistance, Microbial , Erythromycin , Escherichia coli , Escherichia , Mutation Rate , Nalidixic Acid , Parents , Rifampin , TetracyclineABSTRACT
The survival or growth rate of multiple-antibiotic-resistance (Mar) mutants of Escherichia coli K-12 MC4100 or W3110, influenced by many stresses (4% ethanol, 3mM HzO, 0.5M NaC1, pH 2, pH 5, and heat-shock (42C)), and the effects of salicylate (SAL) induction were evaluated by growth rate. Under the stress conditions of pH 5 and heat-shock, growth rates of Mar mutants derived from MC4100 or W3110 induced by salicylate (SAL) were higher than those of wild type Escherichia coli K-12 and E. coli clinical isolates. Under the SAL induction, growth rates of wild type MC4100 or W3110 were higher than those of Mar mutants in case of 0.5M NaC1 or pH 2 stress, respectively. Without SAL induction, growth rates of wild type E. coli were higher than those of Mar mutants, in case of MC4100 grown with 4% ethanol, HO, or heat-shock stresses, in case of W3110 grown with pH 2, or heat-shock stress. There were sorne instances of cytotoxic effect of SAL to the Mar mutants, wild type E. coli, or E. coli clinical isolates under some stress conditions.
Subject(s)
Escherichia coli , Escherichia , Ethanol , Hydrogen-Ion ConcentrationABSTRACT
No abstract available.
ABSTRACT
This is an autopsy case of a 6 month old girl who suddenly died of respiratory distress during sleep. She had suffered from mild but frequent episodes of common cold and was treated for eczema for several days. At autopsy, the heart was enlarged and weighed 100 gm. A firm and gray-white tumor, measuring 4.5 x 3.8 x 2.8 cm, was located in the interventricular septum and encroached upon the wall of left ventricle. The mass was well demarcated but was not encapsulated. Neither necrosis nor calcification was present. Microscopically the tumor was composed of haphazardly arranged bundles of collagen fibers and fibroblasts. Myocardial cells are intermingled with the fibroblasts at the margin of the tumor. Massive edema of the lung and congestion of the liver and spleen were pronounced.
ABSTRACT
Cervical intraepithelial neoplasia in human consist of dysplasia of various developmental stages and squamous cell carcinoma in situ of various types. These lesions can be diagnosed cytologically on cervico-vaginal smears, although the diagnostic reproducibility is limited. To obtain the objects morphologic distinction between normal squamous epithelial cell in different maturation, different stages of dysplastic cells and varieties of in situ carcinoma cells, Kontron IBAS-1 imaging analyzer was applied for the measurement of nuclear and cytoplasmic areas of each categorised cells. The followings are results obtained: 1) Nuclear and cytoplasmic areas of superfical (36.9 micrometer2, 2319.9 micrometer2) intermedicate (45.7 micrometer2, 2989.7 micrometer2) and parabasal cells (50.8 micrometer2, 432.7 micrometer2) of normal squamous epithelium origin are mostly distinctive between cell types. However, cytoplasmic areas of both superficial and intermediate cells and nuclear areas of both intermediate and parabasal cells are not significantly different. 2) Normal squamous cells and various dysplastic cells show obvious difference in their nuclear and cytoplasmic areas, while difference between cytoplasmic areas of both parabasal (432.7 micrometer2) and severe dysplastic cells (409.7 micrometer2) are not statistically significant. 3) No statistical difference is observed in between nuclear areas of both moderate dysplastic (112.3 micrometer2) and severe dysplastic cell (117.6 micrometer2). 4) Varieties of carcinoma in situ cells and severe dysplastic cells are in difference in their nuclear and cytoplastic areas, whereas nuclear areas from both in situ carcinoma cells (95.3 micrometer2) of large cell type and severe dysplasia (117.6 micrometer2) are not distinctive. The results lead the author to consider that the morphometric analysis for various parameters of cell constituents are of value in making objective distinction between cells from cervical intraepithelial neoplasia in human.
Subject(s)
HumansABSTRACT
The authors reviewed surgical materials from 20 patients with carcinoma of the extrahepatic biliary system, and a correlation between macroscopic appearance of the tumors with various clinical features and histopathologic findings was made. Microscopically, the tumors were classified into four types; Four (21%) patients had polypoid tumors, six (32%) had nodular growths, five (26%) were scirrhous constricting in type, and four (21%) had diffusely infiltrating type. Histologically all the differentiation in two cases. The degree of differentiation of the tumors was classified into 3 types: 11 (55%) patients were well differentiated, 3(15%) were moderately well and 6(30%) were poorly differentiated. All polypoid tumors were well differentiated and had low stage. No correlation in the degree of differentiation of the tumor with the stage was present. No correlation in clinical symptoms, duration of symptoms, laboratory findings with morphologic findings of the tumors was noted.