In vitro antimicrobial effect and in vivo preventive and therapeutic effects of partially purified lantibiotic lacticin NK34 against infection by Staphylococcus species isolated from bovine mastitis.

Lantibiotics are small (<5 kDa), polycyclic peptides produced by gram-positive bacteria; they are also known as gram-positive bacteriocins. The high antimicrobial activity of lacticins and the continuing appearance of antibiotic-resistant bacteria in recent years have resulted in a renewed interest in lantibiotics. A partially purified form of lacticin NK34 (a Lactococcus lactis product isolated from the Korean fermented fish jeotgal) was tested to determine its antimicrobial effects against Staphylococcus aureus (n=20) and coagulase-negative Staphylococcus (CNS, n=20) strains isolated from the raw milk of cows with subclinical bovine mastitis in the present study. The spot-on-lawn assay was used to identify the 2 strains from each group with the greatest lacticin NK34 susceptibility, and the minimal lethal dose (MLD) was measured in ICR (imprinting control region) mice. The preventive and therapeutic effects of lacticin NK34 on the mouse infection model were determined for the first time. Lacticin NK34 demonstrated antimicrobial effects in 14 of 20 (70%) S. aureus indicator strains and in 18 of 20 (90%) CNS strains. Staphylococcus aureus 69 and S. simulans 55 demonstrated the greatest susceptibility to lacticin NK34 in the spot-on-lawn assay. The S. aureus 69 MLD was measured at 1.53 x 10(9) cfu/mouse, whereas the S. simulans 55 MLD was 3.59 x 10(9) cfu/mouse. Mice infected experimentally with S. aureus 69 MLD or S. simulans 55 MLD were treated with lacticin NK34. Treated mice demonstrated an 80% survival rate (48 of 60 mice) compared with a survival rate of 7.5% (3 of 40 mice) in control mice treated with distilled water. These data suggest that lacticin NK34 might be useful in the control of bovine mastitis and systemic bacterial infection.

Antiplatelet effects of acidamides isolated from the fruits of Piper longum L.

The inhibitory effects of four acidamides, piperine, pipernonaline, piperoctadecalidine, and piperlongumine, isolated from the fruits of Piper longum L. on washed rabbit platelet aggregation were examined. All of the four tested acidamides showed dose-dependent inhibitory activities on washed rabbit platelet aggregation induced by collagen, arachidonic acid (AA), and platelet-activating factor (PAF), except for that induced by thrombin. Piperlongumine, in particular, showed stronger inhibitory effects than other acidamides to rabbit platelet aggregation induced by collagen, AA and PAF.

Effects of supplementation of beta-glucan on the growth performance and immunity in broilers.

Two experiments were conducted to evaluate the efficacy of beta-glucan on commercial broilers. In experiment 1, one hundred and forty-four broiler chicks were employed in a 2x3 factorial design with cage and open floor housing with three levels of beta-glucan viz. 0%, 0.02% and 0.04%. In experiment 2, ninety-six broilers were used with 4 treatments: No beta-glucan and antibiotic (T1), beta-glucan 0.03% (T2), antibiotic (T3), and beta-glucan 0.03% + antibiotic (T4) for 34 d with 3 replicates of 8 chicks each in both studies. During experiment 1 there was no significant effect of the feeding system or the beta-glucan levels on the performance from 0 to 17 d but during 18-34 days birds housed on the open floor had significantly (p<0.0001) higher weight gain compared with those in cages. In experiment 2, no significant effect was noticed on the weight gains when the effect of beta-glucan, antibiotic or their interaction were tested. The retention of dry matter increased in both experiments with beta-glucan supplementation. The CD8 and TCR 1 cells were significantly higher in the 0.04% beta-glucan group at 42 days as compared with the control. It could be concluded that beta-glucan supplementation was beneficial for broilers.

Application of remote monitoring and automatic control system using neural network for small wastewater treatment plants in Korea.

For this study, an automatic control system has been developed by using a neural network and internet-based remote monitoring system for efficient operation of plants that have a serious variance of influent loading and have difficulties in appropriate maintenance, just like small wastewater treatment plants in Korea. In the control algorithm, ORP was used as the main sensor for control. At the point where the ORP value was judged to reach the "nitrate knee" of denitrification and phosphorus release, ORP indicated the state of lower saturation read by the neural network and then changed the operating condition from the reduction state to the oxidation state. For example, if ORP indicates the state of higher saturation at the point of "nitrogen breakpoint" or "ammonia valley" of nitrification, the neural network reads it and cuts off the oxygen supply and mixing. The dORP data have been used as one of the main input for the neural network. After the operation of lab-scale cyclic aeration process using an automatic control system, it has been found that regardless of loading variance, more than 95% of organic matters and more than 60% of nitrogen and phosphorus have been removed. Assuming that an internet-connected computer and a basic web browser are available, this study has developed a remote monitoring system that can monitor the operating status of small plants or any troubles with them.

Automatic control and remote monitoring system for biological nutrient removal on small wastewater treatment plants in Korea.

Many small-size wastewater treatment plants in Korea's rural communities are designed to remove organic and suspended matter only, and they generally show a large fluctuation in the influent loading compared to municipal wastewater treatment plants (MWWTPs). They also have no professional engineers stationed for efficient operation against mechanical breakdown. For those reasons, the wastewater treatment plants have low efficiency in treatment of nitrogen and phosphorus as well as organic matter. In order to solve those problems, this study developed an automatic control system and RMS (remote monitoring system), which can keep efficiency stable despite any change in the small plants' loading rates and are capable of removing nutrient materials such as nitrogen or phosphorus. According to the results of the Experimental SBR system of the automatic control program, complete nitrification was made under oxic conditions and denitrification occurred as NO3-N concentration decreased by 0.5 mg/l in anoxic conditions and excellent nitrogen removal efficiency was seen generally. The Experimental SBR system created "phosphate release and uptake" effectively and displayed phosphate-removing efficiency up to more than 80% as the concentration of effluent was kept low by 0.4 mg/l. RMS developed in this study transmits a plant's data and operation states to clients in remote locations in real-time interval through the Internet. Therefore, although you are in a remote location, it allows you to see if a plant is properly operated or there is any breakdown.

Protective effects of green tea polyphenol against reactive oxygen species-induced oxidative stress in cultured rat calvarial osteoblast.

The injurious effects of reactive oxygen species on osteoblasts and the potential protective role played by green tea polyphenols (GtPP) were investigated using primarily cultured rat calvarial osteoblasts. Oxidative stress was induced in cultured osteoblasts, either by adding 100 mmol/L H2O2 or by the action of 40 U/L xanthine oxidase (XO) in the presence of xanthine (250 micromol/L). After incubation, the cellular viability, function and morphology were evaluated. Both treatments produced a significant reduction in osteoblast viability, as assessed by a two-colored fluorescence staining method combined with flow cytometric analysis and MTT assay. A significant reduction in the alkaline phosphatase activity was observed after H2O2 addition, whereas XO did not have the same effect. On the microscopic observations, the morphological changes and intracellular ultrastructural damages were remarkably induced by both treatments. The H2O2-induced alterations were prevented by pre-incubating the osteoblasts with 200 microg/ml GtPP for 1 h. When the oxidative stress was induced by XO, the cellular viability and morphology was also maintained at the same polyphenol concentration. These results demonstrate that GtPP can act as a biological antioxidant in a cell culture experimental model and protect cells from oxidative stress-induced toxicity.

Immunostimulatory effects of anionic alkali mineral complex solution Barodon in porcine lymphocytes.

The anionic alkali mineral complex solution, Barodon (Barodon-S.F. Corp., Korea), was evaluated for its effectiveness as a nonspecific immunostimulator in pigs. The effects of Barodon were determined by analysis of feed efficiency, growth rate, and phenotype of leukocyte subpopulations using monoclonal antibodies specific to porcine leukocyte differentiation antigens and flow cytometry (FC). The study was focused to investigate the change in proportion of the CD4+CD8+ double positive T lymphocyte subpopulation (dpp) which exists uniquely in pigs. In addition, the mitogen-stimulated lymphoproliferative response, tissue distribution in lymphoid organs and the adjuvant effect of Barodon on hog cholera vaccine efficiency were determined. The study has revealed the average daily gain rates and feed conversion rates were significantly (p<0.05) improved in either group of pigs fed with 0.05% Barodon-spray feed (Tx-1) or pigs fed with 3% Barodon-fermented feed (Tx-2) in comparison with group of pigs fed with feed containing no Barodon (control). The proportion of cells expressing CD4+ antigen in Barodon-treated group increased from 3 weeks posttreatment and was significantly higher (p<0.05) than that of control at 8 weeks posttreatment. Particularly, the significantly higher proportion was maintained from 8 weeks through 13 weeks posttreatment in Tx-1 group (p<0.05). The proportion of cells expressing CD8+ antigen was significantly higher at 3 weeks posttreatment in Tx-2 (p<0.01). Proportion of MHC class II-expressing cells was significantly higher in Tx-1 and Tx-2 group at 11 weeks and 8 weeks posttreatment (p<0.05), respectively. In addition, the proportion of Non T/Non B (N) cells was also significantly higher in Tx-2 at 3 weeks posttreatment (p<0.01) and maintained to 13 weeks posttreatment (p<0.1). Between Barodon-treated groups, the proportion of MHC class II-expressing cells was observed to be larger in Tx-2 than Tx-1 from 3 weeks to 8 weeks posttreatment (p<0.05). However, there were no significant difference in the proportions of CD2+ cells, B cells, monocytes and granulocytes between Barodon-treated and control group during the experiment. Dual-color FC analysis, study has revealed an increased proportion of dpp present in lymphocytes obtained from peripheral blood (PB) and mesenteric lymph node (MLN) of Barodon-treated group at 8 and 11 weeks posttreatment. The proportion of dpp in PB was 27.5% and 32.1% in Tx-1 and Tx-2, respectively, but only 2.2% in control group at 8 weeks posttreatment. In MLN, the proportion was 45.1% and 52.1% in Tx-1 and Tx-2, respectively, otherwise 16.5% in control group at 8 weeks posttreatment. The mitogen-stimulated activity was significantly higher in Tx-1 than in the control group at 11 weeks posttreatment when cells were stimulated with Con A and PHA, respectively (p<0.01). Also, Con A-, PHA and PWM-stimulated activity was significantly higher in Tx-2 than in the control group at the same time (p<0.05). The tissue distribution of CD4+, CD8+ and CD4+CD8+ dpp in MLN and spleen was significantly larger in Tx-1 and Tx-2 than in the control group (p<0.01). Also, a larger proportion of dpp was observed in Tx-2 than Tx-1 in spleen between Barodon-treated groups (p<0.01). In conclusion, the study has demonstrated that Barodon had an immunostimulatory effect on pigs through proliferation and activation of porcine immune cells, specially CD4+CD8+ dpp lymphocytes.

Relation between lymphocyte subpopulations of peripheral blood and immune responses of modified live hog cholera virus vaccine in pigs treated with an ionized alkali mineral complex.

Thirty-nine healthy pigs (28-32 days old) were purchased from a commercial swine farm and housed at swine pens of the College. The animals were vaccinated intramuscularly (1 ml) with an attenuated live hog cholera virus (HCV, LOM strain) and then boostered at 5 weeks after the first vaccination. The animals were divided into 4 experimental groups: 0.05% (w/w) PowerFeel-supplemented diet (T-1, n = 10); 3% (w/w) SuperFeed-supplemented diet (T-2, n = 10); diluted PowerFeel solution (1 : 500, v/v) as drinking water (T-3, n=9); control (n=10). PowerFeel is an original form of ionized alkali mineral complex (IAMC) and SuperFeed is a commercial product of IAMC. The subpopulation of lymphocyte in blood was assayed by a flow cytometry and HCV-specific antibody was determined by an indirect immunofluorescence assay. In IMAC-treated groups, the proportions of subpopulation expressing MHC-class II, CD2+, CD4+, CD8+, and surface IgM+ B lymphocytes were significantly decreased at 5-weeks after the first vaccination. Significant decreases were also observed in the proportions of MHC-class II, CD2+ and CD8+ lymphocyte at 3-weeks after the booster injection. The humoral immune responses in T-1 and T-2 groups were greater than those in T-3 or control group. These results suggest that IAMC-supplemented diets may have an HCV-specific immunostimulatory effect in pigs.

Solubilization of biphenyl dimethyl dicarboxylate by cosolvency.

Biphenyl dimethyl dicarboxylate (BDD) is a synthetic analogue of schizandrin C, one of the components isolated from Fructus schizandrae, and has been widely prescribed for improvement of liver functions and symptoms of patients with liver disease. However, its oral preparations have been known to have limited bioavailability due to its extremely low solubility in water, and its solubility problem also limits preparation of its parenteral dosage forms. In this research, we searched for solvent systems to solubilize BDD to overcome these problems. The ternary solvent systems of N,N'-dimethylacetamide (DMA)/alcohol/water and Cremophor EL/DMA/alcohol were studied intensively for this purpose. BDD was solubilized effectively in these cosolvents, and the results showed that the cosolvent systems were effective for solubilizing BDD up to the concentration that might be employed for preparation of parenteral dosage forms. Formulation of a BDD concentrate for intravenous infusion was proposed employing the cosolvent system of Cremophor EL/DMA/alcohol.

Reduction of chronic ciclosporin nephrotoxicity by thromboxane synthase inhibition with OKY-046.

Ciclosporin A (CsA) is a potent immunosuppressive agent which is extremely effective in controlling allograft rejection and in the treatment of autoimmune disease and nephrotic syndrome. Unfortunately, its use is limited by chronic, irreversible nephrotoxicity. Administration of CsA induces renal vasoconstriction, causing a reduction in renal blood flow. An alteration of the prostaglandin-thromboxane cascade may be involved in the vasoconstriction. We studied the role of thromboxane A2 in CsA nephrotoxicity and the ability of a thromboxane synthase inhibitor, OKY-046, to reduce the CsA nephrotoxicity. Daily administration of CsA 25 mg/kg for 28 days to Sprague-Dawley rats resulted in increased excretion of urinary thomboxane B2 (47.9+/-11.5 vs. 27.2+/-9.7 ng/24 h; p<0.05) and decreased creatinine clearance (0.25+/-0.07 vs. 0.43+/-0.17ml/min/100 g; p<0.01) as compared with administration of vehicle only. Histologically, large numbers of lysosomes in the tubular epithelium were characteristic. Coadministration of OKY-046 prevented both the rise in urinary thromboxane B2 excretion (40.0+/-11.8 ng/24 h) and the reduction in the creatinine clearance (0.44+/-0.11 ml/min/100 g). The severity of the histological changes was significantly diminished. Selective inhibition of thromboxane production with OKY-046 may be valuable in the attenuation of CsA nephrotoxicity.

A sulfonoglycolipid with Na+,K+-ATPase inhibitory activity, produced by a cultured unique diatom symbiont isolated from a larger foraminifera.

A unique diatom (Entomoneis sp.) was isolated from a larger foraminifera (Marginopora vertebralis) and was successfully mass cultured. 6-Sulfo-O-alpha-D-quinovopyranosyldiacylglycerol was isolated from this diatom as an Na+,K+-ATPase inhibitor.

[Effectiveness of dapsone on refractory immune thrombocytopenia in a patient with systemic lupus erythematosus associated with sarcoidosis].

A 57-year-old man was admitted with massive nasal bleeding and blurred vision in January, 1991. Laboratory examination showed a prominent decrease of platelet number (1,000/microliters) and a marked elevation of PAIgG (4,025 ng/10(7) cells). Serological test revealed positive antinuclear factor, low concentration of C3 and C4, high level of immune complex and polyclonal hypergammaglobulinemia. The patient had uveitis and bilateral hilar lymphadenopathy with a high level of serum lysozyme and negative PPD skin test. The diagnosis of SLE complicated with thrombocytopenia and sarcoidosis was made. In spite of the various trials of treatment, such as oral prednisolone (PSL), methyl-PSL pulse therapy, plasma exchange, high-dose intravenous gammaglobulin, cyclophosphamide, azathioprine, vincristine, colchicine, cyclosporine-A, mizoribine, danazol, ascorbic acid and interferon alpha 2b, the platelet number could not be raised enough to keep more than 10,000/microliters, though the level of PAIgG decreased to 200 ng/10(7) cells. Finally, the administration of 75 mg/day of dapsone brought about a significant rise in platelet number within 2 weeks. The maximum number of 6.2 x 10(4)/microliters was obtained after 2 months. Then the patient stopped himself to take the drug, but the platelet number remained around 4-5 x 10(4)/microliters. Same dose of the drug was again prescribed to confirm the effect of dapsone. The platelet number increased to 7.9 x 10(4)/microliters in 2 weeks, and gradually returned to 5 x 10(4)/microliters after cessation of the drug. Thus being certainly effective against thrombocytopenia, dapsone should be considered as one of the therapeutic choice for refractory autoimmune thrombocytopenia.

Hepatocellular carcinoma with skeletal metastasis: management with intraarterial radioactive iodine.

Two patients with skeletal metastasis from hepatocellular carcinoma (HCC) treated with internal radiation are presented. An oily contrast medium injected into the hepatic artery accumulates in the hepatic tumor tissue and remains there for long periods: therefore, an oily contrast tagged with radioactive iodine could intensify therapeutic effect on tumors. I-131 was tagged onto Lipiodol, an iodized oil, and injected into the arteries supplying the tumors of skeletal metastasis and hepatocellular carcinoma delivering high internal radiation to both primary and secondary lesions. At 6-month follow-up period, tumor has decreased in size and patients are alive and pain free.

Antimicrobial agents from higher plants. Antimicrobial isoflavanoids and related substances from Glycyrrhiza glabra L. var. typica.

Bioassay-directed fractionation of Glycyrrhiza glabra L. var. typica resulted in the isolation and characterization of glabridin (I), glabrol (2), glabrene (3), 3-hydroxyglabrol (4), 4'-O-methylglabridin (5), 3'-methoxyglabridin (6), formononetin (7), phaseollinisoflavan (8), hispaglabridin A (9), hispaglabridin B (13), salicylic acid and O-acetyl salicylic acid. Of these, hispaglabridin A, hispaglabridin B, 4'-O-methylglabridin, glabridin, glabrol and 3-hydroxyglabrol possess significant antimicrobial activity in vitro; hispaglabridin A, hispaglabridin B, 3'-methoxyglabridin, 4'-O-methylglabridin 3-hydroxyglabrol, phaseollinisoflavan, salicylic acid, and O, acetyl salicylic acid are newly found in Glycyrrhiza sp.; and hispaglabridin A, hispaglabridin B, 3'-methoxyglabridin, 4'-O-methylglabridin, and 3-hydroxyglabrol are new to the literature and their structures are proposed herein.

Antimicrobial agents from higher plants. An investigation of Hunnemannia fumariaefolia pseudoalcoholates of sanguinarine and chelerythrine.

The antimicrobial activity seen in extracts of Hunnemmania fumariaefolia S. resides in the alkaloids. The activity seen in the non-polar fractions had been shown by chromatography, physicochemical measurements, and comparisons with authentic samples to be due to apparently artifactual formation of the pseudomethanolates and pseudoethanolates of the alkaloids sanguinarine and chelerythrine, which are abundant in the plant. The enhanced potency of the pseudoalcoholates of sanguinarine over the parent benzophenanthridine is discussed in comparison with the in vitro biopotencies of other analogs prepared for the purpose. It appears that the pseudoalcoholates may be useful prodrugs.