Enhanced wound healing by topical application of ointment containing a low concentration of povidone-iodine.

OBJECTIVE: To investigate the effect of a novel topical wound-healing agent, low-concentration povidone-iodine ointment (LPIO) with a hydrophobic white petrolatum-rich base on skin-wound models in rats and rabbits. METHOD: The therapeutic efficacy of topically applied LPIO was compared to that of standard-concentration povidone-iodine ointment (SPIO) and non-treatment control, using a full-thickness skin-wound model in 24 hairless rats and a full-thickness skin-defect model in rabbit earlobes. The animals were kept under standardised conditions at the Central Research Laboratory of Maruishi Pharmaceutical Co. Ltd. (Osaka, Japan). Therapeutic efficacy was evaluated based on macroscopic wound-size reduction, as well as histopathological and immuno-histochemical examinations. RESULTS: LPIO enhanced wound healing in rat full-thickness skin ulcers, reducing wound size and inflammation, when compared with that in SPIO and non-treatment control. LPIO also markedly improved wound healing in rabbit earlobe ulcers by significantly improving re-epithelialisation, compared with that in SPIO. CONCLUSION: The results of this study suggest that LPIO is a useful topical therapy for ulcerative lesions.

Heat-killed bacillus Calmette-Guérin and Mycobacterium kansasii antigen 85B combined vaccination ameliorates dermatitis in a mouse model of atopic dermatitis by inducing regulatory T cells.

BACKGROUND: Atopic dermatitis (AD) is a recurrent inflammatory skin disease characterized by dominant T-helper (Th) 2 cytokine response. Bacillus Calmette-Guérin (BCG) has been used for preventing tuberculosis, and is regarded as a strong Th1 cytokine inducer. Antigen (Ag) 85B is a secretory protein present in Mycobacterium species that induces Th1 cytokine production. OBJECTIVES: We investigated the effects of combined vaccination of heat-killed BCG (hkBCG) and Mycobacterium kansasii Ag85B in an AD mouse model. METHODS: For the AD model, keratin 14 promoter-derived caspase-1 overexpressing mice (KCASP1Tg) were used. The mice received a combination therapy of hkBCG at age 3 weeks and Ag85B twice weekly for 11 weeks from the 4th week; Ag85B monotherapy from the 4th week; hkBCG monotherapy at the 3rd week; or control saline. Areas of skin lesions, cytokine mRNA expression and serum interleukin (IL)-18 and immunoglobulin (Ig) E levels were analysed. Inducible Foxp3+ regulatory T cells (iTreg), IL-10-producing T cells (Tr1), and interferon (IFN)-γ/IL-4/IL-17-producing T cells were evaluated in the spleen. RESULTS: Saline-treated mice and hkBCG monotherapy mice spontaneously developed severe dermatitis. However, combined therapy with hkBCG and Ag85B significantly suppressed the development of skin lesions and mast cell infiltrations. Elevations of the serum IgE and IL-18 levels were significantly suppressed with combined therapy. Mice treated with hkBCG and Ag85B had a normal number of iTreg in the spleen, and decreased number of both IL-4- and IL-17-producing CD4+ T cells. The effect of Ag85B monotherapy was limited. CONCLUSIONS: Combined vaccination with hkBCG and Ag85B decreases AD skin lesions by inducing regulatory T cells, suggesting that this vaccination is a potent and novel therapeutic strategy for AD.

1,24-Dihydroxyvitamin D3 (tacalcitol) prevents skin T-cell infiltration.

BACKGROUND: 1,24-Dihydroxyvitamin D3 (tacalcitol), a vitamin D(3) compound, has been used to treat T cell-mediated inflammatory skin diseases such as psoriasis, prurigo and vitiligo. The best-known mechanism of action of this compound is inhibition of the abnormal proliferation of keratinocytes and subsequent maturation; however, its effects on skin T-cell recruitment have not yet been evaluated. Cutaneous lymphocyte-associated antigen (CLA), a surface glycoprotein expressed on T cells, plays a critical role in skin T-cell infiltration. We recently reported that 1,25-dihydroxyvitamin D3 inhibits skin infiltration of CD4+ T cells by suppressing CLA expression on T cells. OBJECTIVES: In this study, we investigated the effect of tacalcitol on CLA epitope decoration and on the levels of gut or lymph node homing receptor expression in human T cells. METHODS: We cultured human T cells with tacalcitol and analysed the effect on CLA expression and skin-homing ability, and evaluated glycosyltransferase mRNAs. We also performed an in vivo study using an antigen-dependent delayed-type hypersensitivity (DTH) mouse model and investigated the effect of tacalcitol on skin-infiltrating CD4+ T cells. RESULTS: Tacalcitol downregulated the expression of CLA and, in parallel, the E- and P-selectin ligand function; however, it exerted no effect on other homing receptors. Subcutaneously and intraperitoneally administered tacalcitol downregulated skin infiltration of effector CD4+ T cells in an in vivo DTH mouse model. CONCLUSIONS: These findings suggest that tacalcitol reduces skin inflammation by partially downregulating CLA expression levels.

IL-4/IL-13 antagonist DNA vaccination successfully suppresses Th2 type chronic dermatitis.

BACKGROUND: Atopic dermatitis (AD) is a chronic disease with a Th2-type-cytokine dominant profile. Several cytokines and related peptides have been used for the treatment of AD but they were ineffective because of their limited biological half-life. We have recently developed a highly efficient mouse dominant negative interleukin (IL)-4/IL-13 antagonist (IL-4DM), which blocks both IL-4 and IL-13 signal transductions. OBJECTIVE: To examine the effects of IL-4DM in vivo in an AD model induced by the repeated exhibition of oxazolone (OX). METHODS: Plasmid DNA was injected intraperitoneally to cause an experimental AD-like dermatitis. The effect was evaluated by ear thickness, histological findings, and mast cells counts in the inflamed skin. The plasma IgE and histamine levels were measured. Cytokine production in skin and splenocytes were also analysed. RESULTS: Mice treated with control plasmid developed marked dermatitis with mast cells and eosinophil infiltration, and had increased plasma IgE and histamine levels with a Th2 type splenocyte cytokine profile. Treatment with mouse IL-4 DNA augmented the ear swelling and thickness with an increased dermal eosinophil count, plasma histamine level, and production of splenocyte IL-4. However, IL-4DM treatment successfully controlled the dermatitis, decreased the mast cell and eosinophil count, and suppressed plasma IgE and histamine levels. Splenocytes produced an increased level of IFN-gamma. CONCLUSION: These data showed that the simultaneous suppression of IL-4/IL-13 signals successfully controlled Th2-type chronic dermatitis. IL-4DM DNA treatment is a potent therapy for AD and related diseases.

What causes hidradenitis suppurativa?

Hidradenitis suppurativa (HS)--a rather common, very chronic and debilitating inflammatory skin appendage disorder with a notoriously underestimated burden of disease--has long been a playground for the high priests of nomenclature: Ask a bunch of eminent dermatologists and skin pathologists to publicly share their thoughts on what causes HS, and they will soon get entrenched in a heated debate on whether this historical term is a despicable misnomer. Fortunately, the recently founded Hidradenitis Suppurativa Foundation (HSF; http://www.hs-foundation.org), to which EXP DERMATOL serves as home journal, has broken with this unproductive tradition and has encouraged publication of the current CONTROVERSIES feature. This is exclusively devoted to discussing the pathobiology of this chronic neutrophilic folliculitis of unknown origin. Although traces of terminological bickering remain visible, it does the HS experts in our virtual debate room credit that they engage in a constructive and comprehensive dissection of potential pathogenesis pathways that may culminate in the clinical picture we know under the competing terms HS or acne inversa. These experts sketch more often complementary than mutually exclusive pathogenesis scenarios, and the outlines of a conceivable consensus on the many open pathobiology questions begin to emerge in these CONTROVERSIES. Hopefully, this heralds a welcome new tradition: to get to the molecular heart of HS pathogenesis, which can only be achieved by a renaissance of solid basic HS research, as the key to developing more effective HS therapy.

Cytokeratin expression in subungual squamous cell carcinoma.

Cytokeratin expression in subungual squamous cell carcinoma was investigated in order to evaluate the origin and state of differentiation of the tumour. The tumour nests contained cytokeratin 14, 16 and 17, which were also expressed in the nail bed. Therefore, cytokeratin expression in subungual squamous cell carcinoma may reflect its indolent clinical prognosis.

De novo Ph-negative acute T-lymphoblastic leukemia with BCR gene rearrangement.

We herein report a rare case of Ph chromosome-negative acute T-lymphoblastic leukemia (T-ALL) with a classical BCR rearrangement seen in chronic myelogenous leukemia (CML). There were no clinical or morphologic features to suggest a previous chronic phase of CML. The importance of a full analysis of the BCR gene in the case of adult acute lymphoblastic leukemia (ALL), especially in T-ALL, are discussed.

Keratinizing squamous epithelium associated with Syringocystadenoma papilliferum differentiates towards infrainfundibulum: case report with immunohistochemical study of cytokeratins.

We describe a case of Syringocystadenoma papilliferum (SCAP) with keratinizing squamous epithelium in a 26-year-old female presenting with a dark brown to black nodule on her forehead. After surgical excision, the specimen was examined immunohistochemically using antibodies against cytokeratin (CK) 1, 8, 10, 14, 17, 18 and 19. Within the keratinizing squamous epithelium, CK1, 10, 14 and 17 were present, whereas the other CKs were absent. Based on CK expression, keratinizing squamous epithelium in SCAP seems to differentiate towards the infrainfundibulum.

Monoclonal antibody labeling for cytokeratins and filaggrin in the human pilosebaceous unit of normal, seborrhoeic and acne skin.

The distribution of cytokeratins and filaggrin in human pilosebaceous unit was investigated in specimens obtained from normal (n = 15), seborrhoeic (n = 6), and acne skin (n = 6), using the monoclonal antibodies CK8.12, CK8.13, CK4.62, CK8.60, KL1, PKK2, RPN 1160, and an antibody for filaggrin. The type and amount of cytokeratin content was correlated with the stage of cell differentiation in these three skin types. In all specimens studied the sebocytes. The sebaceous duct cells, and the infundibular cells contained cytokeratins, no clear differences were found between normal, seborrhoeic, and acne skin. During sebocytic maturation the amount and type of cytokeratin content changed gradually and the labeling pattern was partly different compared to the interfollicular epidermal pattern. In the sebaceous duct and the infundibulum, the labeling pattern using KL1, CK8.12, and CK8.13 was similar to that seen in interfollicular epidermis, whereas labeling with CK8.60 and PKK2 was different. These findings indicate that sebaceous duct and infundibular cells express transitional patterns of differentiation between epidermal keratinocytes and sebocytes. Filaggrin was expressed only in some sebaceous duct cells and in infundibular cells. In seborrhoeic and in acne skin, however, the reactivity of antibody to filaggrin was more intense and was already observed in the lower parts of the sebaceous duct and the infundibulum. Although no filaggrin was found in the intermediate cells of the sebaceous duct and the infundibulum in normal skin, these cell types clearly contained filaggrin in seborrhoeic and acne skin.

Heterogeneity of Leukemic Cells with Basophilic Features: Cytochemical, Ultrastructural and Immunophonotypic Analysis of 8 Cases.

We have studied leukemic cells, derived from acute nonlymphocytic leukemia with basophilic features and basophilic crisis of chronic myelogenous leukemia (CML), by cytochemical and ultrastructural examination and analysis of surface markers. Cytochemical results varied from case to case, while the ultrastructural appearances of the granules were different from normal granules. The granules had more delicate granular matrices with or without myelinoid figures, whorled or scroll matrix, multivesicular bodies structures, theta granules, and crystalloid structures. Leukemic cells in all cases had myeloid surface markers with some degree of variability. In addition, they were occasionally positive for lymphoid markers, but not for CD10 and IgE receptors. The present results show that leukemic cells with basophilic features are heterogeneous in their morphology, cytochemistry and surface markers.

The effect of the immune response to Helicobacter pylori in the development of intestinal metaplasia.

AIM: To determine whether cytotoxic Helicobacter pylori antibodies occur in gastric mucosa, and whether these antibodies contribute to the development of intestinal metaplasia. MATERIALS AND METHODS: The number of mononuclear inflammatory cells, which specifically produce immunoglobulin (Ig)G and IgA antibodies, was investigated by using an enzyme-linked immunospot assay and the fraction of mononuclear inflammatory cells determined in gastric biopsy specimens from 34 subjects with H. pylori infection. Assays for the cytotoxicity of H. pylori antibodies were performed on cultured Japanese green monkey kidney (Vero) cells and by in vitro tests. RESULTS: The number of IgG and IgA antibody-producing cells was positively correlated with the degree of inflammation of the gastric mucosa. However, the number of IgG antibody-producing cells was lower in subjects with intestinal metaplasia than in those without. This was not the case for IgA. Significant cytotoxic damage was observed in Vero cells in vitro when incubated in a solution containing the H. pylori IgG antibody, antigen and polymorphonuclear leukocytes. No cytotoxicity was seen with the IgA antibody or with the antigen or polymorphonuclear leukocytes alone. CONCLUSIONS: H. pylori infection is associated with the appearance of immunocompetent mononuclear cells in gastric mucosa. These cells produce H. pylori antibodies of the IgG class which are capable of causing cytotoxic damage in the epithelial cells, obviously through activation of polymorphonuclear leukocytes by antibody-antigen complexes. The occurrence of these cells is inversely related to intestinal metaplasia, suggesting that they may be involved in the processes of epithelial damage leading to the appearance of intestinal metaplasia.

Effects of subminimal inhibitory concentrations of minocycline on neutrophil chemotactic factor production in comedonal bacteria, neutrophil phagocytosis and oxygen metabolism.

Comedonal bacteria, Propionibacterium acnes, P. granulosum and coagulase-negative staphylococci (CNS) seem to play an important initiating role in the inflammatory process by producing neutrophil chemotactic factors. The attracted neutrophils, after phagocytosis, release inflammatory factors such as reactive oxygen species (ROS). We investigated the effects of minocycline at subminimal inhibitory concentrations (sub-MIC), i.e. one-tenth MIC, on the production of human neutrophil chemotactic factors in comedonal bacteria, and on several inflammatory parameters of neutrophils, including neutrophil phagocytosis and generation of ROS (O2-, H2O2, OH.). ROS generation in a cell-free, xanthine-xanthine oxidase system was also assessed. Production of neutrophil chemotactic factors in all strains of P. acnes, P. granulosum and CNS were significantly suppressed by sub-MIC minocycline. Sub-MIC minocycline effectively reduced three kinds of neutrophil-generated ROS (O2-, H2O2, OH.). However, neutrophil phagocytosis and the ROS generated in a cell-free system were not markedly changed in the presence of sub-MIC minocycline. The results suggest that sub-MIC minocycline has an anti-inflammatory effect by inhibiting the production of neutrophil chemotactic factors in comedonal bacteria as well as ROS generated by neutrophils in the inflammatory process of acne.

Antimicrobial susceptibility of Propionibacterium acnes isolated from acne vulgaris.

Systemic and topical antimicrobial treatment for acne vulgaris remains the mainstay method of therapy in Japan. Strains of Propionibacterium acnes (P. acnes) resistant to erythromycin (EM), clindamycin (CLDM), tetracycline (TC), doxycycline (DOXY) and minocycline (MINO) have been reported. The aim of the present study was to examine the antimicrobial susceptibility to 10 currently used antimicrobial agents of 50 strains of P. acnes isolated from acne lesions and identified using a Rap ID ANA II panel. Minimum inhibitory concentrations (MIC) were determined by the agar dilution method according to the criteria of the Japan Society of Chemotherapy. EM, ampicillin (ABPC), and CLDM were the most potent drugs, followed by MINO, nadifloxacin (NDFX), cephalexin (CEX), DOXY, ofloxacin (OFLX), and TC. In terms of the MIC80, EM and ABPC were the most potent, followed by CLDM, NDFX, MINO, CEX, DOXY, OFLX, TC and gentamycin (GM). Although most of the strains used were susceptible to the antimicrobial agents tested, strains of P. acnes resistant (MIC 12.5 mug/ml) to EM (4%), CLDM (4%), DOXY (2%) and TC (2%) were observed. In this study, no strains of P. acnes resistant to MINO were seen, suggesting that oral MINO is the most useful treatment for acne vulgaris with minimal risk of bacterial resistance.

Asymptomatic hyponatremia due to inappropriate secretion of antidiuretic hormone as the first sign of a small cell lung cancer in an elderly man.

A 72-year-old man was hospitalized with asymptomatic hyponatremia. Despite hyponatremia, urinary sodium excretion with urine osmolality exceeding plasma osmolality persisted. Plasma vasopressin levels were high and independent of plasma osmolality during hypertonic saline infusion. Computed tomography of the chest showed enlarged mediastinal and right hilar lymph nodes. Microscopically, a specimen of lymph nodes obtained by biopsy represented vasopressin-producing small cell lung carcinoma. Chemotherapy plus irradiation improved the hyponatremia. Thus, careful evaluation is necessary to determine the cause of hyponatremia disorders in elderly patients.