Characteristics of cytotoxic cells induced by Toxoplasma lysate antigen in mouse spleen.

Spleen cells from Toxoplasma lysate antigen (TLA)-sensitized BALB/c mice showed the strong cytotoxic activity against both natural killer (NK)-sensitive cells (YAC-1 and RL male-1) and NK-insensitive cells (P-815), when incubated with TLA or recombinant human IL-2 (rhIL-2). The increment of TLA concentration in culture medium increased the cytotoxic activity. Treatment of effector cells; spleen cells from TLA-sensitized mice incubated with TLA, with anti-asialo GM1 or anti-Thy-1 plus complement inhibited the cytotoxic activity of effector cells, whereas treatment with anti-mouse Lyt-2.2 serum plus complement had no effect on the cytotoxic activity. Treatment of spleen cells from TLA-sensitized mice with anti-asialo GM1 and/or anti-Thy-1 plus complement inhibited cytotoxic activities of effector cells. These results suggested that spleen cells sensitized with TLA both in vivo and in vitro were asialo GM1 positive and Thy-1 positive, and the majority of cytotoxic cells induced by TLA were similar to lymphokine-activated killer (LAK) cells induced by IL-2.     

Involvement of a host erythrocyte sialic acid content in Babesia bovis infection

Host sialic acid (SA) has recently been suggested to play an important role in erythrocyte (RBC) infection by Babesia spp. The present study attempted to further determine the specific type of SAs important in the RBC invasion. Bovine RBC was found to bear abundant alpha2-3-linked SA residues but not alpha2-6-linked SA in nature, confirmed by flow cytometric analysis of the neuraminidase (Nm)-treated RBCs. Lectin-blot analyses revealed the removal of alpha2-3-linked SAs from the 97-, 33-, and 31-kDa bands by the Nm treatment. Addition of the Nm-treated RBCs into an in vitro culture of B. bovis resulted in a decreased population of the parasitized RBCs. The thin smear samples from the cultures were then observed under a confocal laser scanning microscope after staining with the alpha2-3-linked SA-specific lectin: a selective invasion of B. bovis was found only in the intact RBCs bearing the SAs, but not in the desialylated RBCs. Furthermore, a significant reduction of the parasitized RBCs was also observed in the culture supplemented with exogenous 3'-sialyllactose containing the alpha2-3-linked SAs. However, the complete inhibition of parasite proliferation was not achieved in the culture. These findings indicate that while the alpha2-3-linked SA-dependent pathway is needed for highly efficient invasion of host RBCs by B. bovis, there might also be other potential alternative pathways.     

Evaluation of in vitro and in vivo inhibitory effects of fusidic acid on Babesia and Theileria parasites

Fusidic acid known to has antibacterial, antifungal, and antimalarial activities. Fusidic acid blocks translation elongation factor G gene in Plasmodium falciparum. In the present study, the inhibitory effects of fusidic acid on the in vitro growth of bovine and equine Babesia parasites were evaluated. The inhibitory effect of fusidic acid on the in vivo growth of Babesia microti was also assessed. The in vitro growth of four Babesia species that were tested was significantly inhibited (P < 0.05) by micromolar concentrations of fusidic acid (IC₅₀ values = 144.8, 17.3, 33.3, and 56.25 μM for Babesia bovis, Babesia bigemina, Babesia caballi, and Theileria equi, respectively). Combinations of fusidic acid with diminazene aceturate synergistically potentiated its inhibitory effects in vitro on B. bovis and B. caballi. In B. microti-infected mice, fusidic acid caused significant (P < 0.05) inhibition of the growth of B. microti at the dose of 500 mg/kg BW relative to control group. These results indicate that fusidic acid might be incorporated in treatment of babesiosis.     

Modeling and Evaluating the Performance of River Sediment on Immobilizing Arsenic from Hydrothermally Altered Rock in Laboratory Column Experiments with Hydrus-1D

Large volumes of excavated rock are produced as a result of road and railway tunnel construction in Hokkaido, Japan. Due to the geological condition of this region, these rocks have often undergone hydrothermal alterations, causing them to contain elevated amounts of hazardous elements including arsenic (As). Therefore, these excavated rocks are potentially hazardous waste, and proper disposal methods are required. In this article, performance of unsaturated river sediment on immobilizing As from hydrothermally altered rock is evaluated using laboratory column experiments and Hydrus-1D. The results reveal that the river sediment significantly reduces As migration. Arsenic retarded by river sediment was observed in three patterns. The first was an adsorption onto minerals originally contained in the river sediment. The next pattern was a combination of reduction of As generation by oxidation of As bearing-minerals, irreversible adsorption, and adsorption onto newly precipitated Fe oxy-hydroxide/oxide. The last pattern led to a further depletion of As leached from the rock layer due to a shift in the majority of the As generation mechanism from dissolution to oxidation in combination with a low concentration of oxygen in the rock layer. These patterns were satisfactorily evaluated by a Hydrus-1D model with reversible and irreversible adsorptions. The information from this work is effective in designing and establishing a reasonable technique for the disposal of hydrothermally altered rocks.     

Antimicrobial activity of some N-(arylalkyl)maleimides

A series of N-(arylalkyl)maleimides was prepared for the reaction of maleic anhydride and N-(arylalkyl) amines, and their antimicrobial activities were examined. All compounds exhibited antibacterial activity against gram-positive bacteria such as Bacillus subtilis and Staphylococcus aureus. Almost all compounds exhibited antibacterial activity against the gram-negative bacterium, Escherichia coli, but were inactive against Pseudomonas aeruginosa. Activities against gram-positive bacteria were independent of the nature of the substituent on the benzene ring or the length of alkyl group, but that against gram-negative bacteria was influenced by these parameters. All N-(arylalkyl)maleimides showed activity against yeasts and mycelial fungi.     

In vitro characterization of the inhibitory effects of ketoconazole on metabolic activities of cytochrome P-450 in canine hepatic microsomes

OBJECTIVE: To evaluate the inhibitory potency of ketoconazole (KTZ) on the metabolic activities of isozymes of cytochrome P-450 (CYP) in dogs. ANIMALS: 4 healthy 1-year-old male Beagles. PROCEDURE: Hepatic microsomes were harvested from 4 dogs after euthanasia. To investigate the effects of KTZ on CYP metabolic activities, 7-ethoxyresorufin, tolbutamide, bufuralol, and midazolam hydrochloride were used as specific substrates for CYP1A1/2, CYP2C21, CYP2D15, and CYP3A12, respectively. The concentrations of metabolites formed by CYP were measured by high-performance liquid chromatography, except for the resorufin concentrations that were measured by a fluorometric method. The reaction velocity-substrate concentration data were analyzed to obtain kinetic variables, including maximum reaction velocity, Michaelis-Menten constant, and inhibitory constant (Ki). RESULTS: KTZ competitively inhibited 7-ethoxyresorufin O-deethylation and midazolam 4-hydroxylation; it noncompetitively inhibited tolbutamide methylhydroxylation. Bufuralol 1'-hydroxylation was inhibited slightly by KTZ. The mean Ki values of KTZ were 10.6+/-6.0, 170+/-2.5, and 0.180+/-0.131 microM for 7-ethoxyresorufin O-deethylation, tolbutamide methylhydroxylation, and midazolam 4-hydroxylation, respectively. CONCLUSIONS AND CLINICAL RELEVANCE: In dogs, KTZ at a therapeutic dose may change the pharmacokinetics of CYP3A12 substrates as a result of inhibition of their biotransformation. Furthermore, no influence of KTZ on the pharmacokinetics of CYP1A1/2, CYP2C21, and CYP2D15 substrates are likely. In clinical practice, adverse drug effects may develop when KTZ is administered concomitantly with a drug that is primarily metabolized by CYP3A12.     

Comparison of macrophage scavenger receptor-A knockout mice with wild type ones in the immune response against repeated infestation with Haemaphysalis longicornis

Using macrophage scavenger receptor-A knockout (SRKO) mice, we examined the role of macrophage class A scavenger receptors (MRS-A) on the immune response and acquisition of host resistance against repeated infestation with Haemaphysalis longicornis. Except for one batch of nymphs that infested one of the SRKO (SR-/-) mice and showed no appreciable reduction in body weight, all the other groups of nymphs manifested significant decrease in body weight. Both SR-/- and wild type (SR+/+) mice showed a sustained increase in anti-tick antibody titers, but SR+/+ mice showed significantly higher titers. The IFN-gamma assayed in SR-/- mouse immune sera was substantially less compared with that in SR+/+ mice. Immune sera from SR-/- and SR+/+ mice recognized the 51 and 44 kDa, and 44 kDa proteins, respectively, of the salivary gland antigen. The difference in the level of anti-tick resistance manifested by both groups of mice may be influenced by less efficient trapping and processing of tick antigens by macrophages in mice lacking for the macrophage scavenger receptors, and consequently affected the cascade of Th1 and Th2 responses. We have thus obtained valuable data that strongly infer the role of MSR-A in enhancing host defense against repeated infestation with H. longicornis.     

Differentiation of Toxocara canis and T. cati eggs by light and scanning electron microscopy

In this study, we investigated the morphological identification of Toxocara canis and T. cati eggs on the basis of light and scanning electron microscopic (SEM) observations. T. canis and T. cati eggs used in this study were recovered from the uteri of respective gravid female worms. Measurement of egg size was not helpful in the differentiation of these species, because approximately 90% of eggs measured were of similar size. Using SEM, we were able to differentiate T. canis eggs from T. cati eggs based on their respective characteristic surface structures. Both species have subspherical eggs with markedly pitted surfaces like those of a golf ball, but the surface pitting in T. canis is more coarse than that in T. cati. In this study, however, these differences were not absolute, as 16% of T. canis and 29% of T. cati eggs showed surface pitting that was uncharacteristic of their species. Of the 16% of T. canis eggs that could not be differentiated by surface structure, 3% had pitting resembling T. cati, and the remaining 13% showed intermediate type surface pitting. Similarly, 5% of T. cati eggs resembled T. canis, and 25% of these were of intermediate type. Light microscopic observation yielded results similar to those of SEM, indicating that light microscopy is also a useful tool for the identification of Toxocara eggs.     

牛卵形巴贝斯虫的体外培养

本试验研究了长期在液氮或－80℃冷冻保存虫株－牛卵形巴贝斯虫（B.ovata)的体外培养方法，并探讨了培养条件，试验结果表明：液氮内保存2年之久的牛卵形巴贝斯虫809814号虫株，先经过BO－SCID小鼠体内大量增殖后，在完全埋头液和37℃，5％CO2，5％O2，90％N2培养箱内能够快速增殖，连续培养45天，继代9次，B.ovata在BO－RBC－SCID小鼠体内染虫率可达15％以上，体外培养最高染虫率为6．3％，染虫率5％以上红细胞可作诊断用抗原的制备材料或用液氮（或冷冻）继续保存。     

Sponge-associated fungi from a mangrove habitat in Indonesia: species composition,antimicrobial activity,enzyme screening and bioactive profiling

There is no report of diversity, biological properties and bioactive compounds of sponge-associated fungi from Indonesia’s mangrove ecosystem to date. This study was designed to isolate sponge-associated fungi from a mangrove ecosystem in Mangkang, to screen the antimicrobial and extracellular enzyme properties of the isolates, characterize the biologically promising isolates using molecular approaches, and profile the secondary metabolites using phytochemical and thin-layer chromatography (TLC) analyses. An unidentified sponge that lived in association with mangrove roots was collected from Mangkang. Total of eight associated fungi were isolated from the sponge. Among all isolates, only two fungi SPMKF 1 and SPMKF 6 produced extracellular amylase, another two fungi SPMKF 4 and 5 showed antibacterial activity against MRSA, and only one fungus SPMKF 8 was able to produce extracellular amylase and show antimicrobial activity against ESBL E. coli, Salmonella enterica ser. Typhi strain MDR and C. albicans, while SPMKF 2, SPMKF 3 and SPMKF 9 did not show any biological properties. The result of genetic characterization proved that SPMKF 1 was Cladosporium tenuissimum, SPMKF 4 was Eutypella sp., SPMKF 5 was Lasiodiplodia theobromae, SPMKF 6 was Fusarium keratoplasticum and SPMKF 8 was F. solani. Furthermore, an amylase gene was detected in fungi SPMKF 1, 6 and 8 while among the BGC, only NRPS genes were detected in SPMKF 4, 5 and 8. Interestingly, several same metabolites indicating the same retention factor (Rf) values in TLC were detected in the fungal crude extracts by TLC.