Possible involvement of lymphocyte activating factor (LAF) as an endogenous pyrogen in fever induced by the cell wall skeleton of Nocardia rubra (N-CWS)

We investigated whether interleukin-1 (IL-1) acts as an endogenous pyrogen (EP) on the fever caused by the cell wall skeleton of Nocardia rubra (N-CWS) in guinea pigs. IL-1 activity was expressed as potency of lymphocyte activating factor (LAF). When guinea pig peritoneal macrophages were pulse-stimulated with N-CWS (1-100 micrograms/ml), dose-dependent LAF activity was detected in the supernatants after culture for 4 h. Gel filtration of the culture supernatants on Sephadex G-200 showed that the fractions with LAF activity were not the same as those with cytotoxic activity for L-929 cells, which was measured as an index of tumor necrosis factor (TNF) in parallel with LAF activity. Pyretic activity was detected both in the fractions with LAF activity and in those with cytotoxic activity for L-929 cells. Furthermore, when these macrophages were pulse-stimulated again, this time with the supernatant obtained from macrophages previously pulse-stimulated with N-CWS, LAF and cytotoxic activity for L-929 cells continued to be released from the macrophages. We suggest that IL-1 might be a possible EP in the process of fever elicited by N-CWS, and that such an EP stimulates the macrophages to release further IL-1 or TNF. The resultant long-lasting fever would thus be caused by the continuous release of an EP.     

A possible mechanism of fever induced by Nocardia rubra cell wall skeleton (N-CWS) in experimental animals

The characteristics of fever elicited by the cell wall skeleton of Nocardia rubra (N-CWS) and by lipopolysaccharide (LPS) were compared in rabbits, and the possible involvement of the antigenicity of N-CWS was investigated in guinea pigs. In rabbits, fever of more than 0.5 degree C developed after an intravenous (i.v.) injection of 10 micrograms/kg or more of N-CWS, and was monophasic with 30-100 micrograms/kg but biphasic with the highest dose of 300 micrograms/kg. LPS elicited fever with similar characteristics at doses of 0.01-0.1 microgram/kg. With both compounds, the fever was inhibited by indomethacin. Tolerance to N-CWS and LPS appeared after dosing with 30 or 0.1 micrograms/kg, respectively for 10 d. In guinea pigs sensitized with N-CWS, challenge with 1 or 10 micrograms/kg of N-CWS 10 d later, which did not induce fever in the nonsensitized animals, caused fever of more than 0.5 degree C, and delayed-type hypersensitivity (DTH) appeared. N-CWS also elicited fever in nonsensitized guinea pigs bearing N-CWS-sensitized lymphocytes or anti-N-CWS antibody; the fever was higher in the guinea pigs sensitized with the lymphocytes than in those with the anti-N-CWS antibody. In brief, single injections of N-CWS and of LPS elicited fever with similar characteristics, although the potency of N-CWS was weaker. With N-CWS, the fever is proposed to be triggered by the antigenicity of the compound itself, because doses as low as 1 or 10 micrograms/kg elicited fever along with immunological response in N-CWS-sensitized animals, but not in nonsensitized ones.     

Isolation, characterization and expression of the complementary DNA for human tumor necrosis factor (TNF-alpha)

A cDNA for human TNF-alpha (615bp) was isolated by means of polymerase chain reaction (PCR) using first strand cDNA from PMA-induced HL-60 cells as template. The result from sequencing the 615 bp cDNA fragment indicated that it corresponded to the entire sequence of mature human TNF coding region. Direct expression of mature human TNF was achieved using a plasmid pHT-1 constructed by ligation of the cDNA and a synthetic DNA. The IPTG-induced bacterial product (hTNF) showed cytotoxicity to mouse L-929 cells. The TNF activity was further identified by neutralization of a specific monoclonal antibody against human TNF-alpha. Approximately 80,000 units of activity were detected per ml of culture at A600 = 2.     

聚L-苏氨酸修饰电极对多巴胺和肾上腺素的电催化氧化

利用循环伏安法将L-苏氨酸聚合修饰在玻碳电极表面, 制成聚L-苏氨酸修饰电极. 实验表明, 该电极对多巴胺和肾上腺素都有较好的催化氧化效果. 运用循环伏安法详细研究了修饰电极的电化学性质. 在pH 2.5的磷酸盐缓冲溶液(PBS)中, 肾上腺素的电子传递系数为0.51, 表观反应速率常数为1.33 s-1; 在pH 7.5的PBS中, 多巴胺在电极上产生一对氧化还原峰, 多巴胺在电极上的电子传递系数为0.60, 表观反应速率常数为0.92 s-1. 该修饰电极对多巴胺和肾上腺素能够进行同时测定, 还原峰电流与多巴胺和肾上腺素浓度分别在1.0×10-6-5.0×10-4 mol·L-1和3.0×10-6-1.0×10-4 mol·L-1范围内呈现良好的线性关系.     

Effects of Pigment-Protein Fraction from Nannocloropsis Oculata on TNFα and IL-6 which Act as an Anti-Inflammatory Against Viral Nervous Necrosis (VNN) Infection

The anti-inflammatory effect caused by VNN infection in the grouper fish using pigment-protein fraction of N.oculata has studied. Methods were to explore the pigment-protein fraction of N.oculata, in vivo test, and measurement of anti-inflammatory response using dot blott and immunohistochemistry technique. Results indicate that pigment-protein fraction of N. oculata was able to suppress grouper tissue inflammation when VNN infection was occurring. Inflammation sign was reduced by administration of pigment-protein fraction. This reaction was reinforced by raising of expression of TNFα and IL-6, which means that TNFα and IL-6 acts as an anti-inflammatory in fish tissue.     

Interspecies differences in pharmacokinetics of an antiallergic agent, 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1,4-diazepin-1-yl) benzimidazole difumarate (KG-2413) after intravenous administration to rats, guinea pigs and dogs

Interspecies differences in the pharmacokinetics of an antiallergic agent, 1-(2-ethoxyethyl)-2-(hexahydro-4-methyl-1,4-diazepin-1-yl)benzimid azole difumarate (KG-2413) after intravenous administration were investigated in rats, guinea pigs and dogs. The disappearance of unchanged KG-2413 base was described by biexponential curves in all three animal species. The areas under the plasma concentration-time curves (AUC) in rats, guinea pigs and dogs were 218, 421 and 369 ng.h/ml, respectively, at a dose of 2 mg/kg. The volume of distribution in rats was comparable to that in dogs, and was about three times greater than that in guinea pigs. This might be due to the difference in the unbound fractions of KG-2413 base in plasma (fu), that is, the values of fu in rats, dogs and guinea pigs were 0.607, 0.603 and 0.189, respectively. The first-order elimination rate constant from the central compartment (kcl) in dogs were smaller than those in rats and guinea pigs. Total body clearances (CLtot) of KG-2413 base in rats and guinea pigs were comparable to the hepatic blood flow rate (Q) of each animal. Assuming that KG-2413 base is only eliminated in the liver, relatively rapid metabolism of KG-2413 base occurred in the liver of rats and guinea pigs. In dogs, extrahepatic elimination was suggested because the value of CLtot seemed to be greater than that of Q.     

Solubilization of limulus test reactive material(s) from Candida cells by murine phagocytes.

Solubilization of limulus test reactive materials from Candida was examined in the presence or absence of phagocytic cells. Solubilized limulus test reactive materials (LTRM) were detected in culture supernatant, and hot water and sodium hydroxide extracts of the acetone dried cells of Candida parapsilosis. Suspensions of Candida cells also reacted with limulus test, and LTRM were released from the acetone dried cells by serum treatment. After treatment of the acetone dried cells with polymorphonuclear leucocytes (PMN) or macrophages (M phi), a significant amount of LTRM was solubilized. Significant amounts of LTRM were also released by PMN during treatment of live and growing C. parapsilosis. The reactivity of LTRM was completely inhibited by the addition of excess amount of purified (1----3)-beta-D-glucan, suggesting LTRM from Candida cells as described above would contain (1----3)-beta-D-glucan. These results suggested that LTRM during fungal infection would come from the extracellular water soluble polysaccharide fraction as well as the insoluble cell wall fraction solubilized by the action of phagocytes.     

Fusion Proteins Cpn10-E^rns with Properties of Generating CSFV-Neutralized Antibodies

IntroductionClassical swine fever(CSF), included in the listA of the Office International des Epizooties(OIE), is ahighly contagious disease of domestic pigs and is con-sidered as the mostharmful disease occurring in pigs allover the world[1]. Outbreaks o…     

EFFECTS OF SODIUM SALICYLATE ON THE FEBRILE RESPONSE AND INCREASED LEVELS OF CYCLIC AMP IN CEREBRO-SPINAL FLUID DURING ENDOGENOUS PYROGEN-INDUCED FEVER IN RABBITS

To further evaluate the causality between endogenous pyrogen (EP)-induced fever andcyclic adenosine- 3', 5'- monophosphate (cyclic AMP) level. the effects of sodium salicylate(SS) on the febrile response and increased levels of cyclic AMP in both cerebrospinal fluid(c.s.f.) and plasma during EP- induced fever in rabbits were observed. The results suggestthat cyclic AMP is probably involved in the central mediation of EP-induced fever and thatincreased concentration of cyclic AMP in c.s.f. associated with EP- induced fever is not theresult of temperature elevation but appears to be caused by the increased synthesis in the cen-tral nervous system. In addition it is confirmed that blood is impossibly a contributorysource of increased cyclic AMP in c.s.f. during EP fever, and that SS may act subsequentto the increase in cyclic AMP.     

Soluble glucocorticoid-induced TNF receptor (sGITR) induces inflammation in mice

Glucocorticoid-induced TNF receptor (GITR) was a new member of the TNF/nerve growth factor receptor (TNFR/ NGFR) family and induced in murine T cells by dexamathasone. Recombinant soluble GITR (sGITR) induced an inflammation in peritoneal membrane and changes in spleen after i.p. injection of 3 mg/kg in C57BL/6 mice. Spleen was enlarged and percentage of neutrophils and monocytes were increased. The area of red pulp in spleen was increased, while that of white pulp was decreased after GITR injection. The thickening of membrane and neutrophil infiltration was observed in peritoneal membrane with increased myeloperoxidase activity. At later time, neutrophil infiltration moved to inside the tissue with tissue damage. GITR ligand and GITR were expressed constitutively on the surface of spleen cells and cells from peritoneal fluid. In contrast, no significant change in the spleen and in peritoneal membrane was observed in mice treated with LPS. GITR may play a role in body's inflammatory processes.     

Synthesis,characterization and cytotoxic/cytostatic activity of Sm(III) and Gd(III) complexes

New Sm(III) and Gd(III) complexes of deprotonated 4-hydroxy-3[1-(4-nitrophenyl)-3-oxobutyl]-2H-1-benzopyran-2-one (Acenocoumarol) were synthesized and characterized using FT-IR, FT-Raman, NMR spectra, and elemental analyses. The vibrational study gave evidence for the coordination of ligand to lanthanide ions. The ligand and its lanthanide(III) complexes were tested for their cytotoxic/cytostatic activity against two tumor cell lines and peritoneal mouse macrophages. The Sm(III) and Gd(III) complexes exhibit good activity against melanoma B16 and fibrosarcoma L929 and are stronger inhibitors of tumor cell proliferation than the ligand. Besides their cytotoxicity to tumor cells, Acenocoumarol and its gadolinium(III) and samarium(III) complexes modulate NO generation in activated macrophages.     

Effects of soluble fractions from untreated and SiO2-treated subcellular particles of macrophages on nucleic acid metabolism in isolated nuclei of experimental granulation tissue.

Nuclei isolated from proliferating granulation tissue were incubated with 20 000 g supernatants from untreated and SiO2-treated subcellular particles of rat peritoneal macrophages in the presence of radioactive nucleic acid precursors. The supernatant from SiO2-treated subcellular particles increased the incorporation of [3H]CTP into nuclear RNA maximally by 26% at 5 min, and that of [methyl-3H]dTTP into DNA by 16% at 20 min. The release of radioactivity from labeled DNA was suppressed simultaneously. An RNase preparation from rat peritoneal macrophages enhanced the release of radioactivity from labeled DNA similarly as the soluble fraction from untreated subcellular particles of macrophages. The results suggest that the effects of the soluble fractions upon DNA metabolism of granuloma cells are at least partly independent of the effects on RNA metabolism and that the soluble fraction from SiO2-treated subcellular particles of macrophages stabilizes DNA through inhibition of nuclease activity.     

Intravenously administered (1----3)-beta-D-glucan, SSG, obtained from Sclerotinia sclerotiorum IFO 9395 augments murine peritoneal macrophage functions in vivo.

Effect of intravenously (i.v.) or intraperitoneally (i.p.) administered (1----3)-beta-D-glucan, SSG, obtained from Sclerotinia sclerotiorum IFO 9395 on the murine peritoneal macrophage (PM) functions were examined. A single i.v. administration of SSG increased the number of PMs at a dose of 250 micrograms/mouse, and the peak appeared 4 d after administration. However, no special change was observed on peritoneal exude cell (PEC) populations. These PMs showed augmented lysosomal enzyme activity and the peaks appeared in 2 phases, on days 2 and 10. In contrast, SSG administered i.p. (250 micrograms/mouse) increased the number of PMs and enhanced the lysosomal enzyme activity of PMs from day 4, and a broad peak appeared until days 8--12. The populations of PECs were also changed by i.p. injection of SSG. Additionally, SSG administered i.v. enhanced phagocytic activity, H2O2 production and interleukin 1 (IL-1) production, and the kinetics of the activation differed depending on the activities. These data suggest that the effects of SSG on macrophage functions are different depending on administration routes, and there are some different mechanisms in the activation of macrophages in vivo by SSG.     

REPAIR OF NEAR (365 nm)- AND FAR (254 nm)-UV DAMAGE TO BACTERIOPHAGE OF ESCHERICHIA COLI

Abstract: Intact bacteriophage have been irradiated at 365 nm or at 254 nm and then analysed for DNA photoproducts or injected into their bacterial host to test susceptibility of the damage to both phage and host-cell mediated repair systems. Both thymine dimers and single-strand breaks are induced in the phage DNA by 365 nm radiation. The dimers appear to be the major lethal lesion (approximately 2 dimers per lethal event) in both repair deficient bacteriophage T4 and bacteriophage λ. after irradiation with either 254 nm or 365 nm radiation. Damage induced in T4 by either wavelength is equally susceptible to x -gene reactivation (repair sector approximately 0.5). v -gene reactivation acts on a larger fraction of the near-UV damage (repair sector of 0.82 at 365 nm as against 0.66 at 254 nm). The host-cell mediated photoreactivation system is only slightly less effective for near-UV damage but host-cell reactivation (as measured by comparing survival of phage λ. on a uvr⁺ and a uvr^- host) is effective against a far smaller sector of near-UV damage (0.35) than far-UV damage (0.85). Weigle-reactivation (far-UV induced) of near-UV damage to phage λ is not observed. The results suggest that unless the near-UV damaged phage DNA is repaired immediately after injection. the lesions rapidly lose their susceptibility to repair with a consequent loss of activity of the phage particles.     

DEGRADATION OF THE DNA OF ESCHERICHIA COLI K12 REC- (JC1569b) AFTER IRRADIATION WITH ULTRAVIOLET LIGHT*

Abstract— Degradation of the DNA of a rec^- mutant of Escherichia coli K12 (JC1569 b) induced by u.v. light was investigated. The rate of degradation was much larger by growing bacteria than by stationary cells. When growing bacteria were starved for amino acids, their DNA became resistant to irradiation. The mode of u.v.-induced degradation was investigated by comparing the time course of release from the acid-insoluble fraction of the label for two growing cultures; the one was pulse-labeled with ³H-thymidine and the other was pulse-labeled and chased thereafter for 12 min. It was found that the label incorporated into the former culture begins to be lost from the acid-insoluble fraction prior to the loss of the label incorporated into the latter culture. It was concluded that breakdown of the replicating point precedes degradation of the bulk of the DNA. This result suggested that the replicating point is a sensitive site to irradiation and the u.v.-induced degradation of DNA seemed to be influenced by the state of chromosome at the time of irradiation. Experiments of centrifugation of lysed spheroplasts of bacteria uniformly labeled with ³H-thymidine in alkaline sucrose demonstrated that DNA of low molecular weight appeared after irradiation with only 5 ergs/ mm², and that the molecular weight could not be restored by post-irradiation incubation. Considering these results, an hypothesis is proposed concerning the initiation of induced degradation of the DNA of the rec^- mutant.     

Thin layer chromatographic resolution of some 2-arylpropionic acid enantiomers using L-(-)-serine,L-(-)-threonine and a mixture of L-(-)-serine and L-(-)-threonine-impregnated silica gel as stationary phases

Impregnated silica TLC plates with L-(-)-serine and L-(-)-threonine and a mixture of L-(-)-serine and L-(-)-threonine (1:1) as chiral selectors were prepared to use as chiral stationary phases (CSPs) in thin layer chromatography. The resolution of the enantiomers of 2-arylpropionic drugs, including ibuprofen, ibuproxam, ketoprofen, pranoprofen, benoxaprofen, flurbiprofen and tiaprofenic acid was investigated on these CSPs. A mobile phase system of acetonitrile-methanol-water (16:4:0.5, v/v/v) was used. The spots were detected with iodine vapours and the detection limits were found to range between 0.25 and 0.5 micro g/mL for all racemic compounds investigated. The effect of temperature, pH and concentration of the impregnating chiral selectors on resolution has been studied.     

Cordycedipeptide A, a new cyclodipeptide from the culture liquid of Cordyceps sinensis (Berk.) Sacc

A new cyclodipeptide named as cordycedipeptide A, a new natural compound and two known compound were isolated from the culture liquid of Cordyceps sinensis (BERK.) SACC. Their structures were elucidated as 3-acetamino-6-isobutyl-2,5-dioxopiperazine (1), 3-isopropyl-6-isobutyl-2,5-dioxopiperazine (2) and 3,6-di(4-hydroxy)benzyl-2,5-dioxopiperazine (3) by 1D and 2D-NMR techniques. The cytotoxic assay showed compound 1 had the cytotoxic activities to L-929, A375, and Hela.     

Prevention of TNF-induced necrotic cell death by rottlerin through a Nox1 NADPH oxidase

Previous studies have demonstrated that rottlerin, a specific PKCdelta inhibitor, potentiates death receptor- mediated apoptosis through a cytochrome c-dependent or -independent pathway. However, its ability to regulate necrotic cell death, as well as the underlying mechanism, remains unknown. We found that in murine fibrosarcoma L929 cells, treatment with rottlerin protected the cells against TNF-induced necrosis, whereas it sensitized the cells to apoptosis induced by co-treatment with Hsp90 inhibitor geldanamycin and TNF, in a manner independent of its ability to inhibit PKC-delta. TNF treatment induced rapid accumulation of mitochondrial superoxide (O2-) through the Nox1 NADPH oxidase when cells undergo necrosis. Moreover, pretreatment with rottlerin failed to induce the GTP-bound form of small GTPase Rac1 by TNF treatment, and subsequently suppressed mitochondrial O2- production and poly(ADP-ribose) polymerase activation, thus inhibiting necrotic cell death. Therefore, our study suggests that Nox1 NADPH oxidase is a new molecular target for anti-necrotic activity of rottlerin upon death-receptor ligation.     

Microwaves and cellular immunity. II. Immunostimulating effects of microwaves and naturally occurring antioxidant nutrients

The effect of 8.15-18 GHz (1 Hz within) microwave radiation at a power density of 1 microW/cm2 on the tumor necrosis factor (TNF) production and immune response was tested. A single 5 h whole-body exposure induced a significant increase in TNF production in peritoneal macrophages and splenic T cells. The mitogenic response in T lymphocytes increased after microwave exposure. The activation of cellular immunity was observed within 3 days after exposure. The diet containing lipid-soluble nutrients (beta-carotene, alpha-tocopherol and ubiquinone Q9) increased the activity of macrophages and T cells from irradiated mice. These results demonstrate that irradiation with low-power density microwaves stimulates the immune potential of macrophages and T cells, and the antioxidant treatment enhances the effect of microwaves, in particular at later terms, when the effect of irradiation is reduced.     

Topical trans-4-aminomethylcyclohexanecarboxylic acid prevents ultraviolet radiation-induced pigmentation

We have studied the effect of a plasmin inhibitor, trans-4-aminomethylcyclohexanecarboxylic acid (trans-AMCHA), on skin pigmentation induced by ultraviolet (UV) exposure in Weiser-Maples guinea pigs. When guinea pigs are exposed to UV radiation (840 mJ cm-2), skin pigmentation is clearly observed from seven days after exposure and continued to increase to 29 days. Post-exposure applications of 2 and 3% solutions of trans-AMCHA to the exposed regions prevent or inhibit the pigmentation process. When the skin is removed and stained by the Fontana-Masson method, melanin content in the basal layer of UV-exposed epidermis is significantly reduced in the regions to which 2 and 3% trans-AMCHA solutions have been applied, compared with the vehicle control. As plasmin is known to contribute to the release of arachidonic acid (AA) and the production of prostaglandins (PGs), we have examined the effects of trans-AMCHA on AA-induced pigmentation in guinea pig skin. Topical application of trans-AMCHA causes a dose-dependent decrease in AA-induced pigmentation. These results suggest that trans-AMCHA reduces melanocyte tyrosinase activity by suppressing the production of PGs, UV-induced melanogens, through the suppression of the UV-induced increase in epidermal plasmin activity.