Distribution of kerosene components in rats following dermal exposure.

The systemic distribution of kerosene components in blood and tissues was analysed in rats following dermal exposure. Four types of trimethylbenzenes (TMBs) and aliphatic hydrocarbons (AHCs) with carbon numbers 9-16 (C(9)-C(16)) were analysed as major kerosene components by capillary gas chromatography/mass spectrometry (GC/MS). The kerosene components were detected in blood and all tissues after a small piece of cotton soaked with kerosene was applied to the abdominal skin. The amounts of TMBs detected were higher than those of AHCs. Greater increases in TMB levels were found in adipose tissue in an exposure duration-dependent manner. The amounts of TMBs detected were only at trace levels following post-mortem dermal exposure to kerosene. These findings suggest that kerosene components were absorbed percutaneously and distributed to various organs via the blood circulation. Post-mortem or ante-mortem exposure to kerosene could be distinguished when the exposure duration was relatively long. Adipose tissue would seem to be the most useful for estimating the degree of kerosene exposure.

Comparative roles of free fatty acids with reactive nitrogen intermediates and reactive oxygen intermediates in expression of the anti-microbial activity of macrophages against Mycobacterium tuberculosis.

We assessed the role of free fatty acids (FFA) in the expression of the activity of macrophages against Mycobacterium tuberculosis in relation to the roles of two major anti-microbial effectors, reactive nitrogen intermediates (RNI) and reactive oxygen intermediates (ROI). Intracellular growth of M. tuberculosis residing inside macrophages was accelerated by treatments of macrophages with either quinacrine (phospholipase A2 (PLA2) inhibitor), arachidonyl trifuloromethylketone (type IV cytosolic PLA2 inhibitor), NG-monomethyl-L-arginine (nitric oxide synthase inhibitor), and superoxide dismutase plus catalase (ROI scavengers). In addition, M. tuberculosis-infected macrophages produced and/or secreted these effectors sequentially in the order ROI (0-3 h), FFA (0-48 h), and RNI (3 to at least 72 h). Notably, membranous FFA (arachidonic acid) of macrophages translocated to M. tuberculosis residing in the phagosomes of macrophages in phagocytic ability- and PLA2-dependent fashions during cultivation after M. tuberculosis infection. FFA, RNI and H2O2-mediated halogenation system (H2O2-halogenation system) displayed strong activity against M. tuberculosis in cell-free systems, while ROI alone exerted no such effects. Combinations of 'FFA + RNI' and 'RNI + H2O2-halogenation system' exhibited synergistic and additive effects against M. tuberculosis, respectively, while 'FFA + H2O2-halogenation system' had an antagonistic effect. Moreover, a sequential attack of FFA followed by RNI exerted synergistic activity against M. tuberculosis. Since M. tuberculosis-infected macrophages showed simultaneous production of RNI with FFA secretion for relatively long periods (approx. 45 h) and prolonged RNI production was seen thereafter, RNI in combination with FFA appear to play critical roles in the manifestation of the activity of macrophages against M. tuberculosis.

Increases in mast cells and chymase in fibroproliferative paws of collagen-induced arthritic mice.

OBJECTIVE AND DESIGN: To investigate whether mast cells (MCs) and chymase, the major protease of murine MCs, were involved in a chronic fibroproliferative disorder of the paws associated with type II collagen (CII)-induced arthritis. MATERIALS: Eighteen DBA/1J mice were divided into 3 groups and were used to study fibroproliferative changes in paws elicited by immunization. TREATMENT: Arthritis was induced by immunization with CII, which was intradermally injected as an emulsion made with adjuvant. A booster shot was done 3 weeks after the initial shot. A group with no treatment and that received adjuvant alone served as control. METHODS: Twelve weeks after the booster shot, inflammation of the paws was evaluated for pathological and biochemical indices. Chymase activity was determined with a chromogenic peptide substrate. RESULTS: In CII-immunized group, collagen bundles accumulated around the destructed joints. In accordance with the pathological findings, MC density in the affected paws was increased (154.8+/-13.3/mm2; p<0.05 vs. control) and chymase activity was also increased (29.5+/-2.8 mU/mg protein; p<0.01 vs. control). CONCLUSIONS: The present results demonstrate increases in MCs and chymase in fibroproliferative paws of collagen-induced arthritic mice.

Acanthotic type of eccrine poroma on the arm: report of a case.

A 72-year-old Japanese man with eccrine poroma on the arm is described. To the best of our knowledge, he is the fifth patient with this tumor on the arm reported from Japan. The histopathological type of the tumor of our patient was unique because it was acanthotic; those of the previous patients were all of the intradermal type.

Effects of the Chinese traditional medicine mao-bushi-saishin-to on therapeutic efficacy of a new benzoxazinorifamycin, KRM-1648, against Mycobacterium avium infection in mice.

The Chinese traditional medicine mao-bushi-saishin-to (MBST), which has anti-inflammatory effects and has been used to treat the common cold and nasal allergy in Japan, was examined for its effects on the therapeutic activity of a new benzoxazinorifamycin, KRM-1648 (KRM), against Mycobacterium avium complex (MAC) infection in mice. In addition, we examined the effects of MBST on the anti-MAC activity of murine peritoneal macrophages (M phi s). First, MBST significantly increased the anti-MAC therapeutic activity of KRM when given to mice in combination with KRM, although MBST alone did not exhibit such effects. Second, MBST treatment of M phi s significantly enhanced the KRM-mediated killing of MAC bacteria residing in M phi s, although MBST alone did not potentiate the M phi anti-MAC activity. MBST-treated M phi s showed decreased levels of reactive nitrogen intermediate (RNI) release, suggesting that RNIs are not decisive in the expression of the anti-MAC activity of such M phi populations. MBST partially blocked the interleukin-10 (IL-10) production of MAC-infected M phi s without affecting their transforming growth factor beta (TGF-beta)-producing activity. Reverse transcription-PCR analysis of the lung tissues of MAC-infected mice at weeks 4 and 8 after infection revealed a marked increase in the levels of tumor necrosis factor alpha, gamma interferon (IFN-gamma), IL-10, and TGF-beta mRNAs. KRM treatment of infected mice tended to decrease the levels of the test cytokine mRNAs, except that it increased TGF-beta mRNA expression at week 4. MBST treatment did not affect the levels of any cytokine mRNAs at week 8, while it down-regulated cytokine mRNA expression at week 4. At week 8, treatment of mice with a combination of KRM and MBST caused a marked decrease in the levels of the test cytokines mRNAs, especially IL-10 and IFN-gamma mRNAs, although such effects were obscure at week 4. These findings suggest that down-regulation of the expression of IL-10 and TGF-beta is related to the combined therapeutic effects of KRM and MBST against MAC infection.

Skin mast cell promotion of matrix remodeling in burn wound healing in mice: relevance of chymase.

Inflammation, granulation, and collagen accumulation, which are observed in the wound healing process, occasionally lead to hypertrophic scarring. Several in vitro reports have suggested that skin mast cells (MCs) and their major protease, chymase, participate in the healing process as well as in fibrotic skin diseases. The present study examined the potential involvement of MCs and MC chymase in the healing of burns in mouse dorsal skin. The size of the burn wounds, density of the capillaries, collagen accumulation, MC number, and chymase activity were measured before and 1, 3, 7, and 14 days after burning. The healing process corresponded strongly with MC density and chymase activity in both acute and subacute phases. The maximum decrease in MC number and chymase activity occurred on day 3 when tissue loss due to necrosis was maximal. From day 7 to 14, the burn wounds retracted rapidly accompanied by increases in capillaries and collagen fibers, in correspondence with fast increments in MC numbers and chymase activity at the wound edges. The present results combined with previous in vitro results strongly support the contention that skin MC chymase plays a role in the normal wound healing process, and presumably in dermal fibrotic disorders.

Expression of cathepsin B-like enzyme activity in cell lysate from cultured human normal keratinocytes.

In the present study, we have found that the cell lysate from cultured human normal keratinocytes from foreskin (HFKs) hydrolyzed alpha-N-benzoyl-DL-arginine beta-naphthylamide (BANA), and the BANA hydrolysis occurred most under conditions of 37 degrees C and pH 6.0. This activity was strongly inhibited by leupeptin, which is an inhibitor to cathepsin B. These results suggested that the cell lysate from cultured HFKs contained cathepsin B-like enzyme activity. This is the first report to demonstrate that cathepsin B-like enzyme activity was expressed in the cell lysate from human normal keratinocytes.

Effector molecules in expression of the antimicrobial activity of macrophages against Mycobacterium avium complex: roles of reactive nitrogen intermediates, reactive oxygen intermediates, and free fatty acids.

We studied microbicidal activities of reactive nitrogen intermediates (RNI), free fatty acids (FFA), and reactive oxygen intermediates (ROI) against Mycobacterium avium complex (MAC) and the mode of macrophage (mphi) production of these effectors. (1) Intracellular growth of MAC in murine peritoneal mphis was accelerated by scavengers for ROI or RNI and inhibitors of nitric oxide synthase or phospholipase A2, indicating roles of ROI, RNI, and FFA in mphi anti-MAC functions. (2) Acidified NaNO2-derived RNI, FFA (linolenic and arachidonic acids), and the H2O2-mediated halogenation system exhibited a significant anti-MAC bactericidal activity. The combination of RNI with FFA showed a synergistic effect. However, the H2O2-halogenation system in combination with either RNI or FFA showed an antagonism. When Listeria monocytogenes (Lm) was used as a target organism, the combinations of RNI + FFA and RNI + H2O2-halogenation gave a synergistic effect, whereas FFA + H2O2-halogenation showed an antagonism in exerting bactericidal activity. In addition, when ROI generated by the xanthine oxidase-acetaldehyde system was combined with RNI, anti-Lm but not anti-MAC activity was potentiated. (3) ROI production by murine peritoneal mphis was observed immediately after contact with MAC organisms (MAC stimulation) and ceased within 2 h. FFA release was seen 1-24 h after MAC stimulation. RNI production was initiated from 3 h and increased during the first 36 h and continued at least for 4 days. These findings suggest that RNI and FFA rather than ROI are important effectors of anti-MAC functions of mphis, and the collaborating action of RNI with FFA temporarily participates in mphi-mediated killing of MAC in the relatively early phase after MAC stimulation.

Angiosarcoma which developed on the abdominal wall: report of a case.

We report a 68-year-old Japanese woman with angiosarcoma (AS) on the abdominal wall. To the best of our knowledge, she is the third such patient reported from Japan.

Kaposi's varicelliform eruption in a patient with mycosis fungoides.

A 39-year-old Japanese woman suffering from plaque stage of mycosis fungoides (MF) developed Kaposi's varicelliform eruption (KVE) on her face. KVE appeared 1 month after commencing skin electron beam irradiation (SEBI), when the total irradiation had reached 46 Gy. Natural killer (NK) cell activity in the peripheral blood was abnormally low, but returned to normal after 1 year. However, lymphocyte reactivity to mitogens was persistently low. These facts suggested that the KVE in our patient developed because of abnormally low NK cell reactivity probably induced by radiation therapy.

Inhibitory effect of lead on tube formation by cultured human vascular endothelial cells.

The effect of lead acetate (Pb) on the formation of capillary-like structures (tube formation) by cultured human umbilical vascular endothelial cells (HUVECs) was examined. HUVECs were seeded on a gelled basement membrane matrix (Matrigel). Treatment of HUVECs with 0.3-30.0 microM Pb for 24 hours inhibited the tube formation dose-dependently. The length of tube formation decreased time-dependently with 3.0-10.0 microM Pb. To elucidate the main target factor of Pb for this inhibition, the effects of Pb on the activity of protein kinase C (PKC) and Matrigel were examined. The addition of beta-phorbol 12-myristate 13-acetate (PMA, 50 nM), an activator of PKC, and isoquinolinesulfonamide derivative (H-7, 30 microM), an inhibitor of PKC, showed an increase and decrease in the tube formation, respectively. However, the results of simultaneous addition of Pb and either PMA or H-7 to HUVECs indicated that PMA and H-7 acted not synergistically but additively. When PKC activities in HUVECs were measured by a colorimetric assay after treatments with 3.0-10.0 microM Pb for 24 hours, there was no significant change in PKC activity in the cells. The Pb-inhibition of tube formation was suggested to be independent of PKC activity. Pretreatment of Matrigel with 3.0-10.0 microM Pb for different periods decreased the tube formation dose- and time-dependently. These findings suggest that Pb can inhibit the tube formation by HUVECs dose- and time-dependently and that the inhibitory effect of Pb could be dependent on the degeneration of Matrigel, not on PKC activity.

Prurigo nodularis associated with advanced gastric cancer: report of a case.

In a male Japanese patient, prurigo nodularis (PN) appeared in association with gastric cancer. The cutaneous pruriginous lesions dramatically improved soon after total gastrectomy without any treatment for the skin lesions. Peripheral eosinophilia seen before the operation also rapidly disappeared. These data suggest that some cytokines involved in gastric cancer might have played an important role in the development of PN in our patient.

Monoclonal nonspecific suppressor factor beta (MNSF beta) inhibits the production of TNF-alpha by lipopolysaccharide-activated macrophages.

The monoclonal nonspecific suppressor factor (MNSF), a lymphokine produced by a murine T cell hybridoma, shows a pleiotropic antigen-nonspecific suppressive function. Most recently, a cDNA encoding a subunit of MNSF (MNSF beta) has been isolated and characterized. Recombinant form of MNSF beta (rMNSF beta) inhibits lymphokine functions, as does native MNSF. In this study, we investigated whether rMNSF beta also affects macrophage function in terms of LPS-induced TNF-alpha production by a mouse macrophage cell line, J774. rMNSF beta suppressed the TNF-alpha production in a dose-dependent manner. This suppressive effect was remarkably reduced when rMNSF beta was added after 6 h of LPS stimulation. In addition, enhancement of TNF-alpha production by IFN-gamma was also suppressed by rMNSF beta. The suppressive effect was partly neutralized by the addition of the serine/threonine phosphatase inhibitor, okadaic acid. This finding suggests that serine/threonine protein phosphatases type 1 and/or 2A may be implicated in the mechanism of action of MNSF.

A case of tick bite from Amblyomma testudinarium on the glans penis.

A tick which bit the glans penis of a 40-year-old Japanese man was surgically excised together with the skin where the bite occurred. It was identified as Amblyomma testudinarium through the taxonomical investigation of its morphological characteristics. To our best knowledge, our patient is the sixth tick bite from this species of tick recorded in the Chugoku District of Japan.

In vitro antimycobacterial activity of a new quinolone, levofloxacin (DR-3355).

OBJECTIVE: To examine in vitro antimycobacterial activity of levofloxacin. DESIGN: Minimum inhibitary concentrations (MICs) of levofloxacin for various mycobacterial species were determined by the agar dilution method using 7H11 medium and compared with those of ofloxacin. Antimicrobial activity of levofloxacin against Mycobacterium tuberculosis and M. intracellulare phagocytosed in murine peritoneal macrophages was measured in terms of reducing cell-associated bacterial colony forming units (CFUs). RESULTS: MICs of levofloxacin against M. tuberculosis, M. kansasii, M. marinum, M. scrofulaceum, M. avium, M. intracellulare, M. fortuitum, and M. chelonae were 2 to 4 times lower than those of ofloxacin. Levofloxacin exhibited higher efficacy in reducing bacterial CFUs in macrophages than ofloxacin. CONCLUSION: Levofloxacin possessed more potent in vitro antimycobacterial activities as compared to that of ofloxacin.

Therapeutic effect of KRM-1648 with various antimicrobials against Mycobacterium avium complex infection in mice.

A new benzoxazinorifamycin, KRM-1648 (KRM), was studied for its therapeutic efficacy in combination with other antimicrobials against Mycobacterium avium complex infections in mice. When M. intracellulare-infected (intravenously) mice were given KRM, clarithromycin (CAM), sparfloxacin (SPFX), or ethambutol (EB) each alone or in combination, by gavage, once daily 6 times per week (streptomycin [SM] was given subcutaneously twice per week) from day 1, KRM + CAM exhibited combined efficacy in terms of reducing the incidence of gross lung lesions and the bacterial loads in the lungs and spleens. The addition of either EB or EB + SPFX to KRM + CAM increased the efficacy. Moreover, the multi-drug regimen of KRM + CAM + EB + SPFX or ofloxacin [OFLX]) was more efficacious than rifampicin (RMP) + CAM + EB + SPFX (or OFLX). In M. avium infection, KRM + clofazimine was the most efficacious among two-drug combinations tested followed by KRM + SM. KRM + CAM was considerably less effective against M. avium than against M. intracellulare infection. KRM + EB and KRM+OFLX failed to show such a combined effect.