Minoru Miyasato

FURUNCULAR CUTANEOUS MYIASIS CAUSED BY DERMATOBIA HOMINIS LARVAE EOLLOWING TRAVEL TO BRAZIL

A 62‐year‐old Japanese man had visited Rondonia in central Brazii on May 28, 1994, at which time he suffered insect bites on the left axiila and ieft chest regions. Three bites persisted and became tender and painfui. These areas developed into tender nodules with moderate serous drainage from a central pore. Malaise and an intermediate‐grade fever accompanied the eruption. The tender nodules continued after he returned to Japan on July 7. The diagnosis of furunculosis was made by his family physician, but treatment with oral cefdinir (300 mg per day) and naproxen (600 mg per day) for 2 days and application of ointment containing 0.1% gentamicin sulfate failed to resolve the lesions. The patient complained of a crawling sensation under the skin. Since a maggot was removed from the axillary lesion with the aid of the patients fingers, he was referred to the Dermatology Clinic of the Kurume University Hospital on August 1, for evaluation of parasitic diseases.

Serum Levels of Eosinophil Cationic Protein Reflect the State of In vitro Degranulation of Blood Hypodense Eosinophils in Atopic Dermatitis

In patients with atopic dermatitis (AD), serum levels of eosinophil cationic protein (ECP) have been shown to be a good reflector of disease severity. To elucidate what serum levels of ECP actually reflect, ECP levels in serum and plasma and cytological aspects of blood eosinophils were examined in AD patients (n=27) and compared to healthy subjects (n=12). Significantly elevated levels of serum ECP were noted in AD patients, while plasma ECP were uniformly recorded at nadir levels in both AD patients and normal subjects. In addition to blood eosinophilia, AD patients had significantly increased numbers of hypodense eosinophils (HEo) with morphological characteristics consistent with an activated state. Serum ECP levels strongly correlated with HEo numbers rather than with total eosinophil counts. These results indicate that elevated levels of serum ECP may be a consequence of in vitro degranulation of “activated” HEo, not of ECP supplementation from lesional skin. In addition, the dynamic correlations of eosinophil‐associated parameters (total eosinophil counts, HEo numbers, and serum ECP levels) with AD severity suggest that inflammatory events in lesional skin may be involved in causing not only eosinophilopoiesis in bone marrow, but also development of HEo in the periphery, whose degree in turn may be mirrored in the levels of serum ECP in vitro.

Alteration in the density, morphology, and biological properties of eosinophils produced by bullous pemphigoid blister fluid

SummaryBullous pemphigoid blister fluid (BP-BF) was examined for its effects on the density, morphology, and biological properties of eosinophils. Normodense eosinophils (NEo) were prepared from guinea pig peritoneal exudates by Nycodenz density gradient centrifugation. After culturing with BP-BF, NEo were converted into hypodense eosinophils (HEo) in a time-dependent manner. HEo were morphologically different from NEo in that HEo had spheroidal granules each with a lytic crystalloid core and a significantly increased cell volume. These HEo showed an enhanced antibody-and/or complement-dependent helminthotoxic activity to Schistosoma mansoni larvae, amplified chemiluminescence response to opsonized zymosan, and augmented expression of both FcR+ and CR+. These results suggest that BP-BF contains an activity that may not only induce an eosinophil hypodensity as a consequence of increasing cell volume, but simultaneously enhance an eosinophil cytotoxic potential through augmenting cell-surface receptors and receptor-linked oxidative metabolism. In addition, observed tissue accumulation of this activity suggests that eosinophils may be regulated by their phenotypic change in the skin lesions of bullous pemphigoid and be involved in blister formation.

Eosinophil Phenotypes in Bullous Pemphigoid

Eosinophil phenotypes were investigated in peripheral blood and skin lesions from eight patients with bullous pemphigoid (BP). By Nycodenz density gradients fractionation, blood eosinophils were divided into two phenotypes; normodense (>1.080 g/ml) and hypodense (>1.080 g/ml). Increased numbers of hypodense eosinophils were observed in the blood from all patients with BP. Immunocytochemical observations, using an EG2 monoclonal antibody to react with the secretion form of eosinophil cationic protein (ECP), revealed that EG2 was expressed in 86 ± 3% of hypodense phenotypes and 3 ± 2% of normodense phenotypes. Ultrastructurally, hypodense eosinophils were characterized by numerous spheroidal granules, each with a lytic crystalloid core. These indicate that the hypodense phenotype represents a cell in an activated state. Only eosinophils with immunocytochemical and morphological characteristics similar to hypodense phenotypes infiltrated around the basement membrane zone in involved skin of BP. Furthermore, direct adherence of eosinophils associated with degranulation into basal keratinocytes was seen at the sites of blistering lesions. Bullous fluids contained higher concentrations of ECP than sera as determined by a radioimmunosorbent assay; thus hypodense (activated) eosinophils may directly damage the basal keratinocytes by releasing their granule proteins, subsequently leading to dermo‐epidermal separation.

Eosinophil Chemiluminescence Response to Cytokines and Opsonized Zymosans in Atopic Dermatitis

Chemiluminescence (CL) responsiveness of eosinophils (Eos) to zymosan particles coated with either IgG, C3 or both and eosinophilopoiesis-modulating cytokines was investigated in patients with atopic dermatitis (AD), and an attempt was made to correlate these CL responses with serum levels of eosinophil cationic protein (ECP) which increased in response to the extent of AD severity, but not blood eosinophil counts. A high degree of positive correlation existed between serum levels of ECP and either IgG containing opsonized zymosan- or interleukin-5 (IL-5)-induced CLs. The CL values induced by these two stimuli appeared to be directly correlated with the intensity of each ligand-linked receptor expression. These results suggested that Fc gamma RII-and/or IL-5R-mediated eosinophil activation at the site of lesional skins might implicate in deterioration of cutaneous inflammation, and consequently cause an elevation of serum levels of ECP in AD.

Comparison between famciclovir and valacyclovir for acute pain in adult Japanese immunocompetent patients with herpes zoster.

Famciclovir is a guanine analog antiviral drug used commonly for herpes zoster. Efficacy of famciclovir treatment has been reported to be comparable to valacyclovir treatment. Both of these medications reduce the time to complete cessation of zoster‐associated pain including post‐herpetic neuralgia, as compared to acyclovir. We conducted a multicenter, randomized, open clinical trial in order to evaluate the extent of pain relief afforded by these two antiviral drugs during the acute disease phase of herpes zoster. The study group comprised 86 immunocompetent adult patients suffering from herpes zoster, who were treated with either famciclovir or valacyclovir for 7 days. Of these, 55 patients enrolled in this study within 72 h of the onset of the rash and 31 patients after 72 h of the onset. There was a significant reduction in acute herpes zoster pain with famciclovir on day 7 and at 2–3 weeks in both of these patient groups, while with valacyclovir, there was not significant reduction in pain on day 7. Of patients aged 50 years or older, there was a significantly earlier reduction in pain with famciclovir than with valacyclovir. In addition, a significant reduction in the number of patients with pain was observed as early as days 3–4 with famciclovir treatment as compared with valacyclovir treatment. We conclude that famciclovir was superior to valacyclovir in the relief of acute pain of herpes zoster. Accordingly, famciclovir is recommended for herpes zoster patients with moderate symptoms and a visual analog scale score of under 50 mm.

Purification of human blood eosinophils by a combination method using anti-CD16 monoclonal antibody, immunobeads, and Nycodenz density gradient

A simple method is described for the procurement of human blood eosinophil phenotypes by combining an anti-CD16 monoclonal antibody, immunobeads, and a non-toxic and non-ionic density gradient medium, Nycodenz. The purification depends on the removal of mononuclear cells using a 1.076/1.102 g/ml Nycodenz density gradient, partial removal of neutrophils based on different binding to plastic dishes, interaction of residual neutrophils with immunobeads via an anti-CD16 monoclonal antibody and, finally, extraction of eosinophil phenotypes by sifting the immunobeads-loaded neutrophils through an 1.080/1.102 g/ml Nycodenz density gradient. This method permits simultaneous preparation of highly purified normodense (> 1.080 g/ml) and hypodense eosinophils (< 1.080 g/ml) with reasonable chemiluminescence responses to opsonized zymosans and helminthotoxic activity to opsonized schistosomula corresponding to their own immunocytological properties.

Bullous pemphigoid blister fluids influence the density distribution of eosinophils

Bullous pemphigoid (BP) has been suggested to be an autoimmune inflammatory disease affecting the cutaneous basement membrane zone (BMZ) [7]. Immunologic abnormalities include circulating complement-activating autoantibodies against BP antigen [6, 12]. Current evidence suggests that the inflammatory is initiated and directed at the BMZ by antibody-mediated complement activation [8], and that dermoepidermal separation (DES) is due to BMZ injury by activated leukocytes, particularly eosinophilis or neutrophils [5, 10, 11, 15]. Recent investigations on density gradient centrifugation of peripheral blood leukocytes have shown that eosinophils from eosinophilic patients are less dense than those from healthy subjects [4]. These hypodense eosinophils have higher functional and metabolic activities than normodense eosinophils, so that they are suggested to be in activated state [3, 16]. In BP lesion, if eosinophils are in an activated state, they may become hypodense. The experiment described here was designed to determine whether BP blister fluids (BP-BF) have an effect on eosinophil density. Blister fluids were obtained from four patients with BP (Table 1) and a patient with pemphigus vulgaris, who were diagnosed on the basis of clinical and immunological findings, and from two patients with burn. Suction blisters from normal volunteers were induced using the method o.f Kiistala [9]. BP-BF were heat-inactivated at 56~ for 30 min (hiBP-BF), and dialyzed BP-BF (diBP-BF) were prepared by

Polarization Assay Studies of Human Neutrophil Motility

The polarization assay was used to assess the effects of chemoattractants on neutrophil motility by means of cellular configuration. The addition of N‐formyl‐L‐methionyl‐phenylalanine (f‐Met‐Phe) to a neutrophil suspension from human peripheral blood induced a polarized change in cellular configuration, and the ratio of such transfiguration reached a maximum at 10–8 to 10–6 M of f‐Met‐Phe. The polarized configuration appeared 1 min after addition of f‐Met‐Phe and showed its maximum response at 10 min. The number of polarized cells were decreased after exposure to cytochalasin B or incubating with corticosteroid. The increased response of random polarization was induced in a dose‐dependent fashion by ascorbic acid. Tetracycline HCl did not cause any changes in cell shape at any concentrations examined. This assay can be used to examine patients with depressed neutrophil motility, such as those with systemic lupus erythematosus in the active phase and a severe case of atopic dermatitis.

Immunobead and Density Gradient Purification of Paget Cells: In vitro Studies of Proliferation

Previous studies using primary monolayer cultures of epithelial cells from the involved epidermis of patients with mammary and extramammary Pagets disease investigated whether Paget cells proliferate as other malignant cells do. Although epithelial monolayers from the involved skin were maintained for approximately 45 days, no permanent cell lines were established. The proportion of carcinoembryonic antigen (CEA)‐positive cells did not increase in the long‐term cultures.