H Akiyama

Tohoku University, Japan

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Publications (56)161.19 Total impact

  • British Journal of Dermatology 04/2005; 152(3):586-7. · 3.76 Impact Factor
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    ABSTRACT: Panton-Valentine leukocidin (PVL) is mainly associated with necrotic suppurative lesions, such as furuncles and abscesses in the skin and subcutaneous tissue, but it has also been isolated from patients with community-acquired, severe, necrotizing pneumonia. However, the clinical manifestations of furuncles caused by PVL-producing Staphylococcus aureus and the role of patients' background are not fully understood. We used polymerase chain reaction amplification to test for the PVL gene in 161 strains of S. aureus isolated from suppurative skin lesions. For all PVL gene-positive strains isolated from furuncles, we analyzed cutaneous manifestations, patient background characteristics, and bacteriological markers, including coagulase types, presence of the mecA gene, and toxin profiles, and we compared these results with those for PVL gene-negative strains. PVL genes were detected in 16 (40%) of the 40 S. aureus strains isolated from furuncles, 2 (28%) of the 7 strains isolated from carbuncles, 1 (14%) of the 7 strains isolated from abscesses, and 1 (5%) of the 20 strains isolated from folliculitis. PVL gene-positive S. aureus usually causes multiple (rather than single) furuncles, and such furuncles are usually associated with more-intense erythema around the lesions. PVL gene-positive strains were isolated from young adults without underlying diseases, whereas PVL gene-negative strains were isolated from patients with various systemic complications, including diabetes, leukemia, and autoimmune diseases. PVL gene-positive S. aureus strains are involved in the development of multiple furuncles with more-intense erythema, particularly in healthy young adults. An understanding of the characteristics of furuncles due to PVL gene-positive strains might be useful for preventing the development of the severe systemic infections.
    Clinical Infectious Diseases 03/2005; 40(3):381-5. · 9.37 Impact Factor
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    ABSTRACT: Cadexomer iodine releases iodine (0.9% weight/weight) slowly from beads of dextrin and epichlorhydrin. This preparation is an effective debridement and antiseptic agent for chronic exdudative wounds. The purpose of the present study is to examine the influence of cadexomer iodine against glycocalyx production of Staphylococcus aureus isolated from furuncle lesions on cut wounds in mice using confocal laser scanning microscope (CLSM), and the increase in and glycocalyx production of S. aureus in vitro. In the present study, distinct S. aureus cells and glycocalyx were not detected in the dermis around the cadexomer iodine beads or within those beads, while S. aureus cells encircled by glycocalyx were soaked up by the cadexomer beads and were detected within them in vivo and in vitro. We suggest that cadexomer iodine soaks up S. aureus cells encircled by glycocalyx, directly destroys biofilm structures, and collapses glycocalyx during dehydration, and further, that iodine can subsequently kill S. aureus cells within biofilm. Cadexomer iodine is a promising treatment to clear S. aureus cells within biofilm from skin lesions of exudative or infectious wounds and to prevent wound exacerbation.
    The Journal of Dermatology 08/2004; 31(7):529-34. · 2.35 Impact Factor
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    ABSTRACT: Granulysin is a recently identified antimicrobial protein expressed on cytotoxic T cells, natural killer (NK) cells and NKT cells. It has been shown that granulysin contributes to the defence mechanisms against mycobacterial infection. Superficial microbial folliculitis is a common skin disease. In a previous report, we showed that, as a first line of defence, alpha-defensin, a human neutrophil peptide, and beta-defensin (human beta-defensin-2) were expressed in infiltrating neutrophils and in lesional epidermal keratinocytes, respectively, in superficial folliculitis. As we also observed many infiltrating lymphocytes in lesional dermis, we hypothesized that infiltrating lymphocytes may possess antimicrobial substances, such as granulysin, and play a role in the defence mechanism as a second line of defence. Seven specimens of superficial microbial folliculitis diagnosed clinically and histologically were examined by means of immunohistochemistry. To identify the phenotype of cells expressing granulysin, confocal laser microscopic examination was performed. A dense lymphoid cell infiltrate was observed in pustules, in the perivascular regions. A large number of these lymphoid cells were positive for granulysin. Phenotypically, cells consisted of CD3+ T cells, CD8+ T cells and UCHL-1+ T cells. CD20+ cells and CD56+ cells were not observed. Microscopic examination with a confocal laser showed that the lymphocytes producing granulysin were CD3+ and CD4+ T cells but not CD8+ T cells. We showed that many granulysin-bearing T cells infiltrated affected follicles and perilesional dermis in superficial microbial folliculitis. However, few granulysin-positive lymphoid cells were observed in sterile pustular lesions. Our observations indicated that adaptive immunity such as granulysin, a lymphocyte-produced antimicrobial protein, may play an important role in the cutaneous defence mechanism.
    British Journal of Dermatology 06/2004; 150(5):904-9. · 3.76 Impact Factor
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    ABSTRACT: Streptococcus pyogenes and Staphylococcus aureus are often simultaneously detected from many cases of non-bullous impetigo with atopic dermatitis. Using confocal laser scanning microscopy (CLSM), to investigate formation of S. pyogenes microcolonies in skin lesions. The S. pyogenes cells in the stationary growth phase alone were strongly stained with fluorescein isothiocyanate-concanavalin A (FITC-ConA), and this staining was reduced by pretreatment with amylase. Although the components of sugars in glycocalyx produced by S. pyogenes cells are unknown, we suggested that the materials stained by FITC-ConA were consistent with the presence of ConA-reactive sugars in glycocalyx produced by S. pyogenes cells. S. pyogenes cells associated with streptococcal impetigo skin and croton-oil inflamed mouse skin formed microcolonies encircled by materials (glycocalyx) that stained strongly with FITC-ConA, and these findings were consistent with those in biofilms. In croton-oil inflamed mouse skin, polymorphonuclear leukocytes (PMNs) infiltrated to just below the epidermis in the cefdinir-treated group but only to the middle dermis in the cefdinir-non-treated group. In this case S. pyogenes and S. aureus cells formed separate microcolonies and existed independently in the outer walls of pustule lesions of streptococcal impetigo. In skin infections, S. pyogenes and S. aureus formed aggregates of microcolonies (similar to that in biofilms) encircled by glycocalyx, which can make the infection hard to eradicate using an antimicrobial agent alone. The effect of conventional antimicrobial agents against biofilm is mainly due to the increase of the invasion of PMNs into the biofilm.
    Journal of Dermatological Science 10/2003; 32(3):193-9. · 3.52 Impact Factor
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    ABSTRACT: Glycocalyx collapses during dehydration to produce electron-dense accretions. Confocal laser scanning microscopy (CLSM) may be used to visualize fully hydrated microbial biofilms. Using CLSM, to analyse glycocalyx production by Staphylococcus aureus cells in skin lesions of bullous impetigo, atopic dermatitis and pemphigus foliaceus. A second objective was to compare numbers of S. aureus cells in tissue sections prepared by different methods for routine light microscopy. S. aureus cells in skin lesions of impetigo, atopic dermatitis and pemphigus were stained with safranin, and positive staining with fluorescein isothiocyanate-conjugated concanavalin A was considered to indicate the presence of glycocalyx. All S. aureus cells tested in skin lesions of impetigo, atopic dermatitis and pemphigus were covered with glycocalyx and formed microcolonies. The numbers of S. aureus cells in a routine light microscopy section were significantly lower than those in a frozen section that had not been dehydrated with ethanol. S. aureus cells generally produce glycocalyx in skin lesions of bullous impetigo, atopic dermatitis and pemphigus foliaceus, which accounts for the difficulty of removing S. aureus cells from these skin lesions. The glycocalyx may collapse during dehydration and most of the S. aureus cells may be carried away during preparation of routine light microscope sections.
    British Journal of Dermatology 04/2003; 148(3):526-32. · 3.76 Impact Factor
  • British Journal of Dermatology 02/2003; 148(1):188-91. · 3.76 Impact Factor
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    ABSTRACT: Bacteria that adhere to damaged tissues encase themselves in a hydrated matrix of polysaccharides, forming a slimy layer known as a biofilm. This is the first report of detection of glycocalyx production by Staphylococcus aureus using confocal laser scanning microscopy (CLSM) on damaged skin tissues. To analyse glycocalyx production by S. aureus cells on damaged skin tissues and the influence of polymorphonuclear leucocytes (PMNs) and various antimicrobial agents on its production using CLSM in cyclophosphamide (Cy)-treated (neutropenic) or non-Cy-treated (normal) mice. S. aureus cells were inoculated on damaged skin tissues in neutropenic or normal mice with or without topical application of antimicrobial agents. S. aureus cells were stained with safranine, and positive staining with fluorescein isothiocyanate-conjugated concanavalin A was considered to indicate the presence of glycocalyx. All S. aureus cells tested on damaged skin tissues formed microcolonies encircled by glycocalyx. The colony counts of S. aureus cells on croton oil dermatitis in normal mice treated with 2% fusidic acid ointment were about 100 times lower than those in neutropenic mice (control). As S. aureus cells can generally produce a biofilm on damaged skin tissues, antimicrobial agents may not eradicate S. aureus cells without the help of PMNs. S. aureus glycocalyx may play a crucial role in colonization and adherence to damaged skin tissues.
    British Journal of Dermatology 12/2002; 147(5):879-85. · 3.76 Impact Factor
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    ABSTRACT: Defensins are cationic antimicrobial peptides with a broad spectrum. Recently human beta-defensin 2 (hBD-2) has been isolated from psoriatic skin; however, its exact localization and fate have not been fully understood. We studied the distribution pattern of hBD-2 in skin tissues of psoriasis and other inflammatory skin diseases. In the upper spinous and granular layer of psoriasis vulgaris hBD-2 was present in the cytoplasm. In the horny layer the positive signals were in a basket-weave pattern, indicating possible accumulation of hBD-2 in the intercellular space. The similar pattern of hBD-2 distribution was observed in the lesions of nummular eczema and atopic dermatitis. hBD-2 was not detected in the section of normal elbow and knee skin. When isolated psoriatic scales were stained, hBD-2 was detected in a wrapping paper-like distribution pattern surrounding the corneocytes. In horny layer of psoriatic skin hBD-2 was closely associated or colocalized with elafin, which is known to be in extracellular space, as demonstrated by double staining. Western blot analysis using cultured human keratinocytes detected hBD-2 with an expected size in the conditioned medium and in the cell lysates when stimulated with 5% FCS or IL-alpha. These results indicate that hBD-2 was synthesized and remained in cytoplasm in the upper spinous and granular layer, and then secreted into intercellular space in the horny layer. This dynamic change in hBD-2 distribution in epidermis is certainly relevant to function as an innate host defense mechanism against invading micro-organisms.
    Journal of Molecular Medicine 11/2002; 80(10):678-84. · 4.77 Impact Factor
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    ABSTRACT: Heavy colonization of atopic dermatitis (AD) with Staphylococcus aureus is well documented. The purpose of the present study is to examine the actions of gluco-oligosaccharide (G-OS) against S. aureus for the control of AD skin lesions infected with S. aureus. The colony counts of S. aureus cells in 0.5% sodium chloride solution supplemented with 5% G-OS (pH 4.8) were about 10-fold lower than those in 0.5% sodium chloride solution (pH 6.6; control) after incubation for 24 hours. The colony counts of S. aureus cells attached on the coverslips (pre-treatment with 1% and 5% G-OS/PBS and following treatment with plasma) were about 10-fold lower than those on the coverslips (pre-treatment with PBS and following treatment with plasma; control) in PBS after incubation for 24 hours. The materials (sugars, probably glycocalyx) that stained positively for fluorescein-isothiocyanate (FITC) -concanavalin A and were consistent with the presence of S. aureus cells were reduced when S. aureus cells attached to the coverslips treated with 5% GC-OS. In conclusion, C-OS is a promising agent that can be applied topically in a cream to clear adherent S. aureus cells from skin lesions of AD in order to prevent its exacerbation. Further, 5% C-OS can inhibit glycocalyx production by S. aureus cells and consequently have some suppressive effect on the colonization of S. aureus on the horny cells of AD lesions.
    The Journal of Dermatology 10/2002; 29(9):580-6. · 2.35 Impact Factor
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    ABSTRACT: Human neutrophil peptide-1 (HNP-1), a defensin with antimicrobial properties, is also thought to promote wound healing. To elucidate the mechanism by which wound healing is facilitated by this factor, we investigated the effect of HNP-1 on the expression of interstitial collagenase (matrix metalloproteinase 1, MMP-1), collagen types I and III, and tissue inhibitor of metalloproteinase 1 (TIMP-1) by cultured fibroblasts by means of RT-PCR and ELISA. Our results showed that synthetic HNP-1 increased the expression of proalpha1(I) collagen mRNA and protein. In contrast, the expression of MMP-1 was decreased at both the mRNA and protein levels. Our observations suggest that HNP-1 may promote wound repair by enhancing extracellular matrix deposition and by controlling its degradation.
    Archives for Dermatological Research 08/2002; 294(4):185-9. · 2.71 Impact Factor
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    ABSTRACT: Heavy colonization of atopic dermatitis (AD) with Staphylococcus aureus is well documented. The isolation rate of methicillin-resistant S. aureus is high in strains from AD in Japan. Our objective in the present study was to investigate the actions of farnesol and xylitol against S. aureus for the control of AD skin lesion-colonizing S. aureus. We examined the actions of farnesol on plasma coagulation and superantigenic exotoxin production by S. aureus, the antimicrobial activity of beta-lactam antibiotics combined with farnesol at concentrations below the minimal inhibitory concentration (MIC) and the effect of xylitol on glycocalyx production. Coagulation by S. aureus cells was inhibited in plasma containing farnesol at a concentration of 1/12 of the MIC (100 microg/ml) after incubation for 24 h. The production of superantigenic exotoxins by S. aureus cells with farnesol (100 microg/ml) was about 10 times lower than that by S. aureus cells alone. The MICs of ampicillin and cefdinir against S. aureus were reduced to < or =0.06 microg/ml in Mueller-Hinton agar plates with farnesol (100 microg/ml). We suggest that farnesol at concentrations above the MIC had a suppressive effect against S. aureus cells in the exponential and stationary phase and acted on the cell wall of S. aureus cells in both phases. Farnesol is a promising adjuvant agent against S. aureus skin infections treated with beta-lactam antibiotics. Further, 5% xylitol inhibited glycocalyx production by S. aureus cells and consequently had a suppressive effect on the colonization of S. aureus on the horny cells of AD lesions.
    Chemotherapy 07/2002; 48(3):122-8. · 2.07 Impact Factor
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    ABSTRACT: We used a scanning confocal laser microscope to study the effects of various agents on sugar production by Staphylococcus aureus in vitro. S. aureus cells attached to coverslips in Pl-TSB (plasma:tryptic soy broth=1:1) were stained with fluorescein isothiocyanate-conjugated concanavalin A (FITC-conA) and were more strongly stained over time. We considered that the materials that stained positive for FITC-conA consistent with S. aureus cells were sugars, probably glycocalyx, produced by the S. aureus cells. Since the cells in the stationary growth phase alone were strongly stained with FITC-conA, all S. aureus cells attached to the coverslips in Pl-TSB were considered to be in this phase (low growth rate). The positive staining for FITC-conA was markedly reduced when fibrin was not formed in Pl-TSB with plasmin and sucrose, and was also markedly reduced when the fibrin in Pl-TSB was destroyed with plasmin. In conclusion, the results of the present study indicate that the existence of fibrin is essential for glycocalyx production and biofilm formation of S. aureus cells to aid in the attachment of S. aureus cells in vitro, because S. aureus cells attached on coverslips and fibrin alone produce glycocalyx. Of the antimicrobial agents tested, sulfadiazine silver most strongly inhibited the production of FITC-conA-positive materials by S. aureus cells at a sub-MIC concentration. Plasmin, sucrose, and sulfadiazine silver may be useful topical applications for use on clinical dermatology for the prevention and the treatment of staphylococcal biofilms. We consider that this simple method is very useful for the detection of S. aureus glycocalyx on dermatology field.
    Journal of Dermatological Science 06/2002; 29(1):54-61. · 3.52 Impact Factor
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    ABSTRACT: We examined the antibacterial action of several tannins on plasma coagulation by Staphylococcus aureus and the effect of conventional chemotherapy combined with tannic acid below the MIC. Coagulation was inhibited in plasma containing tannic acid (100 mg/L), gallic acid (5000 mg/L), ellagic acid (5000 mg/L), (-)-epicatechin (1500 mg/L), (-)-epicatechin gallate (500 mg/L) or (-)-epigallocatechin gallate (200 mg/L) after incubation for 24 h. All tannins inhibited coagulation at a concentration below the MIC. The MICs of oxacillin and cefdinir for S. aureus were reduced to < or = 0.06 mg/L in Mueller-Hinton agar plates with tannic acid (100 mg/L) at a concentration below the MIC. The antistaphylococcal activity of tannic acid was reduced in plates with 10% rabbit blood, but not in those with 10% rabbit plasma. Membranous structures formed in a culture medium containing equal proportions of plasma and tryptic soy broth after incubation for 24 h. The colony counts of S. aureus in membranous structures in the medium containing oxacillin (40 mg/L) and tannic acid (100 mg/L) were c. 10-fold lower than those in medium containing oxacillin (40 mg/L) alone (P < 0.01). Tannic acid merits further investigation as a possible adjuvant agent against S. aureus skin infections treated with beta-lactam antibiotics.
    Journal of Antimicrobial Chemotherapy 10/2001; 48(4):487-91. · 5.34 Impact Factor
  • O Yamasaki, H Akiyama, Y Toi, J Arata
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    ABSTRACT: We examined the effects of a combination of roxithromycin and imipenem on a biofilm model of Staphylococcus aureus. Treatment with roxithromycin alone and imipenem alone did not decrease the number of viable bacterial cells compared with the control. However, a combination treatment of roxithromycin and imipenem significantly decreased the number of viable bacterial cells on day 8 after inoculation in the in vivo model (P < 0.01). On days 5 and 8 after inoculation, numerous polymorphonuclear leucocytes and macrophages invaded the bacterial clusters in the roxithromycin- and roxithromycin/imipenem-treated groups, but did not invade the control or imipenem-treated groups. The present study indicated that a combination of roxithromycin and imipenem is a potentially effective treatment for S. aureus biofilm-associated skin infections as it can induce the invasion of polymorphonuclear leucocytes into the biofilm.
    Journal of Antimicrobial Chemotherapy 10/2001; 48(4):573-7. · 5.34 Impact Factor
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    ABSTRACT: The present study examined the antimicrobial effects of acidic hot-spring water on Staphylococcus aureus strains isolated from atopic dermatitis (AD) patients. Plasma coagulation by S. aureus cells was not detected in plasma containing acidic hot-spring water (60%, pH 5.4) or hydrochloric acid (pH 5.0) after incubation for 24 h. S. aureus cells did not grow in Mueller-Hinton broth with acidic hot-spring water (50%, pH 4.4) after 24 h incubation. The colony counts of S. aureus cells in tryptic soy broth containing acidic hot-spring water (60%, pH 3.9) were over ten times lower than those in tryptic soy broth alone after incubation for 24 h (P<0.01). A membranous structure (an immature biofilm) was formed on the coverslips of tissue culture dishes by S. aureus cells in plasma after incubation for 24 h, although the colony counts of S. aureus cells in the immature biofilms in plasma containing acidic hot-spring water (60%, pH 5.4) were about eight times lower than those in plasma alone after incubation for 24 h (P<0.01). The colony counts of S. aureus cells that attached on coverslips in plasma containing acidic hot-spring water (60%, pH 5.4) or hydrochloric acid (pH 5.4) were over 1000 times lower than those in plasma alone after incubation for 24 h. These results suggest that 50% acidic hot-spring water has a bacteriostatic effect, 60% acidic hot-spring water has a moderate bactericidal effect against floating S. aureus cells and those cells in a biofilm, and, 60% acidic hot-spring water has an inhibitory effect on plasma coagulation and attachment of S. aureus cells. Furthermore, our present results suggest that a small amount of some ions in hot-spring water such as manganese and iodide ions are very important for a bactericidal activity of hot-spring water as well as the low pH condition.
    Journal of Dermatological Science 12/2000; 24(2):112-8. · 3.52 Impact Factor
  • H Akiyama, O Yamasaki, J Tada, J Arata
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    ABSTRACT: We examined the production of superantigenic exotoxins in 136 coagulase-negative staphylococci isolated from various skin lesions in humans using a reversed passive latex agglutination test (Denka Seiken). As a control we examined the same in 50 Staphylococcus aureus strains isolated from non-infective skin ulcers in humans. Of the 136 strains of coagulase negative-staphylococci, 9 (6.6%) produced one or more identifiable exotoxins. In contrast, 21 (42%) out of the 50 S. aureus strains produced one or more identifiable exotoxins (P<0.01).
    Journal of Dermatological Science 11/2000; 24(2):142-5. · 3.52 Impact Factor
  • H Akiyama, O Yamasaki, J Tada, J Arata
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    ABSTRACT: We examined the adherence characteristics and susceptibility to various antimicrobial agents of 130 strains of Staphylococcus aureus isolated from infective skin lesions and 135 strains of S. aureus isolated from non-infective eczematous lesions of atopic dermatitis (AD) patients. The isolation rate of methicillin-resistant S. aureus (MRSA) was 27.7% in strains from clinical sources excluding AD and 31.1% in those from AD. Coagulase type II strains were most frequently observed in MRSA strains isolated from all sources excluding AD, and coagulase type III strains were most frequently observed in those isolated from AD. We proposed that antimicrobial treatment for AD patients should be carefully designed to prevent MRSA infection. Plasma coagulation ability was lowest in S. aureus strains isolated from abscesses, suggesting that the lower production of fibrin observed in abscesses may assist the infiltration of neutrophils into skin tissues and that a decrease in plasma coagulation ability may enable abscess formation. Adherence to polypropylene tubes with slime production was most evident in S. aureus strains isolated from felon and least evident in those isolated from cellulitis and lymphangitis. Tube adherence was characteristic of the S. aureus strains attached to superficial skin tissues, but not necessarily for strains that had infiltrated the deep skin tissues. Fusidic acid demonstrated significant antimicrobial activity against the MRSA strains, but rifampicin was the strongest antimicrobial agent.
    Journal of Dermatological Science 09/2000; 23(3):155-60. · 3.52 Impact Factor
  • H Akiyama, O Yamasaki, J Tada, J Arata
    Clinical and Experimental Dermatology 07/2000; 25(4):353-4. · 1.33 Impact Factor
  • H Akiyama, O Yamasaki, J Tada, J Arata
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    ABSTRACT: We examined the effect of ceramic powder slurries on the coagulation of plasma by Staphylococcus aureus cells. Plasma coagulation by S. aureus strains or their cultured supernatant was inhibited in the plasma with 0.12% calcium oxide or 0.25% magnesium oxide after incubation for 24 h at 37 degrees C. Inhibition of plasma coagulation by calcium oxide and magnesium oxide was observed at the lower concentration than zinc oxide.
    Journal of Dermatological Science 01/2000; 22(1):62-5. · 3.52 Impact Factor