Epiglottic diffuse B-cell malignant lymphoma: A case report.

A 55-year-old male patient was admitted to our department with complaints of dysphagia and throat soreness for 2 months. A tumor of the left epiglottis, with an irregular surface, was identified by video laryngoscopy. The diagnosis of malignant lymphoma was confirmed by biopsy during laryngomicrosurgery. The atypical diffuse lymphocytic lymphoma was positive for CD20 and Bcl-2, and negative for CD3, CD10 and Bcl-1. The diagnosis was diffuse large B-cell malignant lymphoma. The patient was treated with eight cycles of rituximab with cyclophosphamide + doxorubicin + vincristine + prednisolone (R-CHOP regimen). This is a rare case of extranodal non-Hodgkin lymphoma occurring in the epiglottis.

Pectinesterase inhibitor from jelly-fig (Ficus awkeotsang Makino) achenes reduces methanol content in carambola wine.

Crude pectinesterase (PE) inhibitor (PEI) extracted from jelly-fig achenes (JFA) (Ficus awakeosang Makino) was added to carambola (Averrhoa carambola L.) puree to determine the change in methanol production during fermentation. Addition of pectin or microbial pectic enzyme to puree increased dose-dependently the methanol content in fermented products. Decreasing ratio (from 1:0 to 1:19, v:v) of pectic enzyme to diluted crude PEI solution in the puree-enzyme mixture decreased the PE activity remarkably. Except for transmittance (%T), addition of crude PEI to puree did not affect apparently the physical and chemical properties of wine; however, it reduced methanol content in the control from 256 to 58 ppm. The degree of esterification (DE) of pectin in starting puree was approximately 70%. It decreased to approximately 27% in the control group and reduced slightly to approximately 67% in fermented puree with crude PEI added after 14 days of fermentation. This reveals that crude PEI solution was potent in inhibiting intrinsic carambola PE activity and appeared to be a potential alternative for methanol reduction in wines.

Novel cross-linked alcohol-insoluble solid (CL-AIS) affinity gel from pea pod for pectinesterase purification.

Alcohol-insoluble solids (AIS) from pea pod were cross-linked (CL-AIS) and used as an affinity gel matrix to isolate pectin esterases (PEs) from tendril shoots of chayote (TSC) and jelly fig achenes (JFA), and the results were compared with those isolated by ion-exchange chromatography with a commercial resin. CL-AIS gel matrix in a column displayed poor absorption and purification fold of PE; however, highly methoxylated CL-AIS (HM-CL-AIS), by exposing CL-AIS to methanolic sulfuric acid to increase the degree of esterification (DE) to 92%, facilitated the enzyme purification. The purified TSC PE and JFA PE by the HM-CL-AIS column were proofed as a single band on an SDS-PAGE gel, showing that the HM-CL-AIS column was a good matrix for purification of PE, either with alkaline isoelectric point (pI) (TSC PE) or with acidic pI (JFA PE).

Pectinesterase inhibitor from jelly fig (Ficus awkeotsang Makino) achene induces apoptosis of human leukemic U937 cells.

The antitumor activity of pectinesterase inhibitor (PEI), a group of cationic polypeptides, from jelly fig (Ficus awkeotsang Makino) achene was first examined as a treatment for leukemia in this study. PEI displayed strong growth inhibition against human leukemic U937 cells via induction of apoptosis in a dose- and time-dependent manner. At a level of 50 microg/mL, PEI inhibited 90% of cell growth, and the concentration of PEI required to induce 50% of cell viability (LC50) was about 180 microg/mL. Meanwhile, cell cycle arrest at G2/M phase was observed when cells were incubated with 100 microg PEI/mL for 24 h. PEI displayed a dose-dependent influence on mitochondria transmembrane potential (MTP, delta psi m) of cells when detected by a flow cytometry. MTP of more than 50% cells was reduced when cells were incubated with PEI at levels higher than 50 microg PEI/mL for 24 h. In addition, PEI upregulated caspase-3 activity. Taken together, PEI potently inhibited the proliferation of human leukemic U937 cells via cell cycle arrest and apoptosis in association with MTP reduction and caspase-3 activation, respectively, and showed therapy potential for U937 cells.

The antiproliferative and differentiating effects of human leukemic U937 cells are mediated by cytokines from activated mononuclear cells by dietary mushrooms.

Proliferation of human leukemic U937 cells was remarkably inhibited by conditioned medium (CM) of human peripheral blood mononuclear cells (MNC-CM) stimulated with cold-water extracts (CWE) (10-800 microg/mL of medium) of dietary mushrooms, Hypsizigus mamoreus (HM), Agrocybe aegerita (AA), Flammulina velutipes (FV), whereas insignificant results were observed when cells were cultured in the presence of CWE at the corresponding level. Water extracts from mushrooms were fractionated by Sephadex G-50 chromatography, and the pooled high molecular weight fraction (F1) (200 microg/mL) of HM (HM1) and AA (AA1) exhibited growth inhibitions >80% on U937 cells. Interestingly, the thus-cultured U937 cells showed high nitroblue tetrazolium (NBT) positive (>68%) and nonspecific esterase (NSE) positive (>47%) percentages, revealing the remarkable differentiation into monocytes/macrophages upon incubation with HM1- and AA1-stimulated MNC-CM. In addition, assays for the expressions of monocyte-associated antigens, CD11b, CD14, and CD68, also evidenced the remarkable differentiation of U937 cells into monocytes/macrophages by presenting high CD maker positive percentages (>50%). Tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta in CM of HM1-stimulated MNC for 1 day (MNC-CM-1) were 1350 and 1374 pg/mL, respectively, revealing the potent antitumor and differentiation-inducing activities of HM. Of note, MNC-CM-1 appeared to be more effective than day 5 MNC-CM (MNC-CM-5) in both antitumor and differentiation-inducing activities.

Simultaneous quantification of glyphosate, glufosinate, and their major metabolites in rice and soybean sprouts by gas chromatography with pulsed flame photometric detector.

Procedures were developed for the simultaneous determination of glyphosate [N-(phosphonomethyl)glycine] and glufosinate [dl-homoalanin-4-yl-(methyl)phosphinic acid] and their major metabolites, aminomethylphosphonic acid (AMPA) and 3-(methylphosphinico)propionic acid (3-MPPA), in rice and soybean sprouts by gas chromatography (GC) equipped with a pulsed flame photometric detector (PFPD). Herbicides and their major metabolites were previously derivatized with TMOA (trimethyl orthoacetate (TMOA) in the presence of acetic acid, and their GC responses versus heating temperature (70-90 degrees C) and heating time (30-120 min) were optimized. It was found that increases in heating temperature and heating time were unfavorable for the derivatization of glyphosate or glufosinate, whereas high temperature and extended reaction time remarkably facilitated that of AMPA and 3-MPPA except at 90 degrees C for an extended reaction time (120 min). Combination of AG1-X8 anion-exchange chromatography with a Florisil cartridge cleanup process was favorable for the GC-PFPD analysis. Four types of derivatives spiked in rice and soybean sprout matrices were eluted, reaching a baseline separation, in a sequence of 3-MPPA, AMPA, glyphosate, and glufosinate within 14 min using a DB-608 capillary column. Recoveries of glyphosate, AMPA, glufosinate, and 3-MPPA (0.5 ppm) spiked in both sample matrices were determined to be 72-81, 71-86, 101-119, and 83-90%, respectively, whereas the coefficient of variation was determined to be <10% in three repeated determinations. The instrumental limits of detection for glyphosate, AMPA, glufosinate, and 3-MPPA in sample matrices were 0.02, 0.03, 0.02, and 0.01 ppm, respectively. The limits of quantification for glyphosate, AMPA, glufosinate, and 3-MPPA in sample matrices were 0.06, 0.10, 0.06, and 0.04 ppm, respectively.

Antiproliferative and differentiating effects of polysaccharide fraction from fu-ling (Poria cocos) on human leukemic U937 and HL-60 cells.

Poria cocos (PC), Fu-Ling, is an oriental fungus and has been widely used as a Chinese traditional herbal medicine for centuries. In the present study, a neutral polysaccharide fraction from PC (PC-PS) was isolated by a series of chromatographies and its effects on antiproliferation and differentiation of human leukemic cells, U937 and HL-60, were investigated in vitro. Results showed that the molecular weight of isolated PC-PS was approximately 160 kDa, as estimated by gel permeation chromatography. The conditioned medium prepared with PC-PS (15 microg/ml)-stimulated human blood mononuclear cells for 5 days (PC-PS-MNC-CM5) exhibited a potent activity in suppressing the proliferation of U937 and HL-60 cells by 87.3 and 74.7%, respectively. Furthermore, PC-PS-MNC-CM5 induced about 66.6% of the U937 cells and 49.4% of the HL-60 cells to differentiate into mature monocytes/macrophages, which also markedly expressed surface antigens of CD11b, CD14, and CD 68. The differentiated U937 and HL-60 cells displayed physiological functions such as respiratory burst and phagocytosis, while PC-PS group and control group showed insignificant effects. Interestingly, the levels of interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha in PC-PS-MNC-CM were about 41 and 10 times, respectively, higher than that of control group. Antibody neutralization tests of the PC-PS-MNC-CM5 revealed that the growth-inhibitory and differentiation-inducing activities were mainly due to the elevated cytokines of IFN-gamma and TNF-alpha. It suggests that PC-PS is a biological response modifier (BRM), instead of a cytotoxic reagent, and may be a potential alternative in leukemia therapy.

Transacylation and de-esterification reactions of pectin as catalyzed by pectinesterases from tomato and citrus.

The optimal conditions for the de-esterification reaction of tomato pectinesterase (PE) and citrus PE was 0.1-0.2 M NaCl and at pH 7.5-8.5, 65 degrees C, almost identical to those for the transacylation reaction as observed by turbidity (absorbance at 400 nm) change. Among the PEs tested, pea pod PE presented the most remarkable catalysis on the transacylation reaction, and 1.5% pectin solution was determined to be suitable for this reaction. Low methoxy pectin with a DE (degree of esterification) of 31% displayed a slow turbidity increase, revealing that the extent of DE was influential on the transacylation. Besides citrus pectin, apple pectin was also proved to progress transacylation reaction by PEs from tomato and citrus sources as apparently observed by turbidity method.

Changes in molecular weight of transacylated pectin catalyzed by tomato and citrus pectinesterases as determined by gel permeation chromatography.

The changes in molecular masses of pectin in 0.5% pectin-pectinesterase (PE) mixtures (2 units/mL) incubated at various temperatures, pH values, and NaCl levels for 30 min were observed by a Toyopearl TSK HW-65 (F) gel permeation chromatography. The molecular mass of pectin was remarkably increased from 103 to 266 kDa when the incubation temperature of pectin-tomato PE was increased from 25 to 45 degrees C. A further increase in molecular mass was observed when a pectin-citrus PE mixture was incubated at 65 degrees C. The values of pH and NaCl levels were also crucial to the transacylation activity of PEs. Reaction at pH 7.5 with tomato PE and citrus PE remarkably expanded the molecular mass of pectin to 410 and 670 kDa, respectively. The NaCl level of 0.3-0.5 and 0.3 M was favorable for the transacylation reaction of tomato PE and citrus PE, respectively. Only high methoxylpectin was the suitable substrate for PE to conduct the transacylation reaction.

Microencapsulation and modification of synthetic peptides of food proteins reduces the blood pressure of spontaneously hypertensive rats.

Synthetic peptides were microencapsulated into liposomes, cycled with a disulfide bond or modified with d-phenylglycine (d-phg) at the N-terminal, and their antihypertensive effects as orally administered (0.18 mM/kg body weight) to spontaneously hypertensive rats (SHR) were measured. The microencapsulated Leu-Lys-Pro reduced significantly the systolic blood pressures of SHR by 45 mmHg and showed a prolonged duration, revealing the significant protective effect of encapsulation. d-phg-Leu-Arg-Pro showed a duration about 2 h shorter than that of the peptide without modification. In addition, cyclic Leu-Arg-Pro peptide with a disulfide bond between the N- and C-terminal amino acids reduced the systolic blood pressure of SHR by 35 mmHg and displayed a lengthy duration.

Isolation and characterization of immunoglobulin in yolk (IgY) specific against hen egg white lysozyme by immunoaffinity chromatography.

Six hens were intramuscularly (im) immunized once a week for 3 weeks using chicken egg white lysozyme (LS) as antigen. Antibody (immunoglobulin in yolk, IgY) ELISA values of 10(3)-fold diluted yolk were almost as high as 1.879 in the sixth week and maintained a value of 0.756 in the eighth week after the initial immunization treatment. The purification efficiency (specific activity of purified IgY against LS/specific activity of antibody in yolk against LS) of IgY specific against LS isolated by laboratory-prepared LS-bound (IgY-) Sepharose 4 Fast Flow immunoaffinity column was approximately 3380. By applying various amounts (0-22 mg) of the thusly obtained IgY specific against LS to the immunoaffinity column, the binding capacity (q(m)) and dissociation constant (K(d), M(-1)) of such immunoaffinity gel for IgY against LS were found to be 0.68 mg of IgY/mL of wet gel (0.54 mg of IgY/mg of LS) and 7.13 x 10(-6) M, respectively, as determined by Langmuir-type adsorption isotherms.

Characterization of pectinesterase inhibitor in jelly fig (Ficus awkeotsang Makino) achenes.

Pectinesterase inhibitor (PEI) extract prepared from intact jelly fig (Ficus awkeotsang Makino) achenes was separated by membrane (MWCO 3 and 10 kDa) and fractionated by a Sepharose G-50 gel permeation chromatography. Results from Sepharose G-50 gel permeation chromatography and concanavalin A Sepharose chromatography revealed PEI as polypeptides with molecular weights ranging from 3.5 to 4.5 kDa. Incubation of a PE (1 unit/mL)-PEI (2 mg/mL) mixture for 1 min decreased the PE activity by approximately 50%. On the basis of the results of Lineweaver-Burk double-reciprocal plots, Michaelis constant (K(m)) and V(max) values for jelly fig achenes PE (pH 6.0, 30 degrees C) were 0.78 mM -OCH3 and 1.09 microequiv of -COOH/min, respectively. In addition, PEI competitively inhibited both citrus and jelly fig achenes PEs.