Prenatal exclusion of harlequin ichthyosis; potential pitfalls in the timing of the fetal skin biopsy.

BACKGROUND: Harlequin ichthyosis (HI) is a severe and usually fatal congenital skin disorder with autosomal recessive inheritance. Several cases of HI prenatal diagnosis have been performed using fetal skin biopsy, mainly at around 23 weeks estimated gestational age (EGA), and reported in the literature. However, prenatal testing must be done earlier than 21 weeks EGA in several countries including Japan where the present HI families live, because termination is legally allowed only until 22 weeks EGA. OBJECTIVES: We report the successful prenatal exclusion of HI in two fetuses from two independent families and discuss the technical difficulties and potential pitfalls in the prenatal exclusion of HI at early gestation stages. METHODS: Fetal skin biopsy specimens and amniotic fluid samples at 19 and 20 weeks EGA from two fetuses at risk of HI were examined by light and electron microscopy. RESULTS: For the prenatal diagnosis in case 1, the fetal skin biopsy samples were obtained at 20 weeks EGA and showed normal keratinization in the hair canals; no abnormalities were observed in the keratinized cells. In case 2, the interfollicular epidermis and the hair follicles in the samples obtained at 19 weeks EGA had not differentiated enough to show proper keratinization. However, lamellar granules were normally formed in the inner root sheath cells of the late bulbous hair pegs. From these ultrastructural findings, the case 1 fetus was diagnosed as unaffected with HI, and the case 2 fetus was diagnosed as unlikely to be affected. Subsequently, both were born as healthy, unaffected babies. CONCLUSIONS: The timing of biopsies at 19 weeks EGA is not ideal for fetal skin biopsy because the samples are not always sufficiently differentiated for the prenatal diagnosis of HI. However, morphological observations of lamellar granules gives us important additional information useful for HI prenatal diagnosis.

Systemic lupus erythematosus-like autoimmune abnormalities induced by bacterial infection.

Recent evidence has revealed that bacterial DNA can promote several of the autoimmune abnormalities observed in systemic lupus erythematosus (SLE), and a possible pathogenic role in the induction of SLE has been highlighted. We have recently encountered patients in whom bacterial infection (septicemia) triggered the production of several autoantibodies. This seems to be interesting with respect to the consideration of the relationship between SLE and bacterial infection.

Allergic diseases in systemic lupus erythematosus: prevalence and immunological considerations.

We attempted to obtain a deeper understanding of the relationship between systemic lupus erythematosus (SLE) and allergic diseases through comparative studies. Accordingly, we reviewed the association of both disorders and compared their immunological features based on the literature and our own findings. Recent studies (including ours) have indicated that the risk of IgE-mediated and/or associated allergic diseases is not markedly increased in SLE patients despite their more allergic family history when compared with controls, in contrast with earlier studies. This may be related to a change of the environmental factors contributing to allergy. In addition, assessment of the immunological similarities and differences between SLE and various allergic diseases seems to be useful for understanding the relationship between them.

Allergic disorders in systemic lupus erythematosus: prevalence and family history.

The relationship between allergic disorders (such as atopic dermatitis, asthma, allergic rhinitis and allergic conjunctivitis) and systemic lupus erythematosus (SLE) is still unclear and controversial. We investigated the prevalence of these allergic disorders in SLE patients and their families. A questionnaire about the history of allergy was completed by 52 SLE patients and by 52 matched (including race, age, sex and region) non-SLE controls. Our results indicated that there was a significantly lower incidence of these allergic diseases in SLE patients, especially those who had an allergic family histories, when compared with the controls. These findings may be related to the immunological similarities and differences between SLE and various allergic diseases.

Sweet's syndrome associated with encephalitis.

The involvement of the central nervous system (CNS) in Sweet's syndrome (acute febrile neutrophilic dermatosis) is rare. We report a 47-year-old woman who presented with acute encephalitis and was subsequently diagnosed as having Sweet's syndrome. She developed altered consciousness following fever and erythematous skin plaques in the extremities. Cerebrospinal fluid (CSF) examination disclosed neutrophilic pleocytosis without decreased glucose level. Brain magnetic resonance imaging (MRI) showed abnormal signal intensity lesions in the basal ganglia and the hippocampus. Skin biopsy revealed a dense dermal infiltration of neutrophils, which is compatible with Sweet's syndrome. Treatment with acyclovir and antibiotics failed, but the subsequent corticosteroid therapy was effective. Awareness of neurological complication in Sweet's syndrome may avoid unnecessary empiric therapy for meningoencephalitis and will lead to a successful treatment with corticosteroids.

Semenogelin, the main protein of semen coagulum, inhibits human sperm capacitation by interfering with the superoxide anion generated during this process.

Semenogelin (Sg), the major protein of the human semen coagulum, is present at high concentrations in seminal vesicle secretions. It is degraded by the prostate-specific antigen (PSA) to generate peptides of various biological activities that were found on and inside spermatozoa. Our aim was to determine the effect of Sg on capacitation, which is the series of transformations that spermatozoa must undergo to become fertile. At concentrations of 0.1 to 1.0 mg/mL (600- to 20-fold lower than those of semen), Sg did not affect sperm motility (%) but completely prevented capacitation induced by fetal cord serum ultrafiltrate; a partial inhibition of capacitation was noted with 0.03 mg Sg/mL. There was also a dose-dependent decrease in the tyrosine phosphorylation of fibrous sheath proteins and in the O2-.-related chemiluminescence. Ribonuclease (RNase), which has as high an isoelectric point (pI = 9.7) as Sg (pI = 9.5), also prevented sperm capacitation and O2-.-related chemiluminescence but to a lower extent, suggesting that one mechanism of Sg action on spermatozoa could be related to its positive charge at physiological pH. Sg at 1, but not 0.3 or 0.1 mg/mL, scavenged the O2-. generated by the mix of xanthine + xanthine oxidase and modified the kinetics of the reaction; RNase did not have such effects. Therefore, Sg is a potential scavenger for O2-. but probably also affects the sperm oxidase. Spermatozoa rapidly processed Sg; a high proportion of Sg was degraded after 15 minutes of incubation. The resulting polypeptide patterns were reminiscent of those obtained with PSA as a proteolytic enzyme. These data suggest that Sg, its degradation products, or both may be natural regulators of sperm capacitation and could prevent this process from occurring prematurely. One mechanism by which Sg acts could involve an interference with the O2-. that is normally generated during this process.

[Induction of atopic dermatitis-like skin lesion in NC/Nga mice--the influence of the skin barrier destroying solution to the induction of dermatitis].

NC/Nga mouse is well known as a mouse model for atopic dermatitis. In general, when NC/Nga mouse are raised under specific pathogen free (SPF) conditions no skin lesions are detected, but when under non-filtrated (conventional) condition, atopic dermatitis like skin lesions appear spontaneously. However, this dermatitis develops in 70-90% of mice (not 100%), which makes it difficult to perform reproducible experiments every time. This study was performed under SPF conditions, using the four solutions (2% SDS, 4% SDS, ethanol, acetone/ether) to destroy the skin barrier function, and thereafter, applying the extracted solution of mite: Dermatophagoides pteronyssinus, which is a very popular antigen in pathogenesis of human atopic dermatitis. The extracted solution of mite was applied repeatedly on the NC/Nga mice with a pretreatment of barrier destroying solution and after 8 weeks the mice developed severe dermatitis (clinical skin condition score of 7-10.2 points) with marked elevation of plasma IgE level, whereas mice coated only with the barrier destroying solution showed weak skin lesion with no elevation of plasma IgE level. BALB/c mice, which are employed as control, showed weak skin lesion (clinical skin condition score of 0-3.8 points) and slight elevation of plasma IgE level after repeated application of the extracted solution of mite with a pretreatment of the barrier destroying solution, whereas mice coated only with the barrier destroying solution showed weak skin lesion and the no elevation of plasma IgE level was observed. In this study, using several solutions to disturb the skin barrier function before applying the antigen, we have found a suitable condition and types of solutions in inducing dermatitis in NC/Nga mice.

A case of antiepiligrin cicatricial pemphigoid successfully treated by plasmapheresis.

We report the case of a 73-year-old Japanese woman suffering from antiepiligrin (laminin 5) cicatricial pemphigoid (CP) with typical clinical and immunopathological features. Histologically, the lesional mucous membrane showed a subepidermal blister formation. When indirect immunofluorescence techniques with skin split by 1 M NaCl as the substrate were used, the patient's serum reacted only to the dermal side. Immunoprecipitation studies demonstrated that the patient's serum contained IgG autoantibodies directed against a set of polypeptides that corresponded to epiligrin (laminin 5). After corticosteroids and immunosuppressive agents had been administered systemically, the patient's autoantibody titer decreased and the cutaneous and mucosal blister formations were suppressed. However, the ocular lesions persisted in spite of these therapeutic regimens. After combining these treatments with double-filtration plasmapheresis, the ocular lesions improved and showed almost no progression. Plasmapheresis may thus present a new option for the treatment of CP.

A case of ichthyosis linearis circumflexa successfully treated with topical tacrolimus.

We report a case of ichthyosis linearis circumflexa (ILC) without the typical atopic manifestations and deformities of the hair shaft. The patient responded positively to treatment with topical tacrolimus, suggesting that abnormalities in the immunoregulatory mechanism may be involved in the pathogenesis of ILC.

[The effect of suplatast tosilate on immunological parameters for the patients with atopic dermatitis].

A dose of 300 mg/day of suplatast tosilate was administered to one hundred one cases of atopic dermatitis for eight weeks, and the severity scores, peripheral blood eosinophil count, total serum IgE levels, plasma eosinophil cationic protein (ECP) levels, and other immunological parameters before and after the trial were observed and comparatively examined. The results are as follows: 1) Temporary improvements were found in the scores of severity and itchiness on all evaluated skin regions (face, limbs, and trunk). These scores decreased significantly for all observation periods at two, four, six and eight weeks after administration of suplatast tosilate compared with those before the administration (p < 0.01). 2) There was no sign of adverse effects on the drug. In the blood tests, one patient displayed elevated levels of GPT and another showed elevated total bilirubin. In the urine test (qualitative test), one case with positive urinary protein was observed. 3) Clinical examinations including assessment of the immunologic parameters were conducted at an average of 8.68 +/- 0.36th week. The peripheral blood eosinophil count, the percentage of eosinophil, and plasma ECP levels significantly diminished compared with those before administration, but no significant difference was found in total serum IgE levels and LDH levels. 4) The subjects were divided into two groups, one in which the clinical scores were improved by more than five and another with scores of less than five (including worsening), and the fluctuation of the immunological parameters (values before and after administration of the drug) of the two groups were compared. As a result, a significant difference was observed in the plasma ECP levels (p = 0.02) and peripheral blood eosinophil count (p = 0.091), but no difference was observed in total serum IgE levels and LDH levels. From the above mentioned results, the high efficacy and safety of suplatast tosilate in the treatment of severe atopic dermatitis were confirmed. At the same time, a decrease in the peripheral blood eosinophil count and the serum ECP levels were observed, suggesting the possibility that these values could be used as indices of the severity of atopic dermatitis.

Antibody raised against extracellular proteinases of Sporothrix schenckii in S. schenckii inoculated hairless mice.

Sporothrix schenckii produces two extracellular proteinases, namely proteinase I and II. Proteinase I is a serine proteinase, inhibited by chymostatin, while proteinase II is an aspartic proteinase, inhibited by pepstatin. Studies on substrate specificity and the effect of proteinase inhibitors on cell growth suggest an important role for these proteinases in terms of fungal invasion and growth. There has, however, been no evidence presented demonstrating that S. schenckii produces 2 extracellular proteinases in vivo. In order to substantiate the in vivo production of proteinases and to attempt a preliminary serodiagnosis of sporotrichosis, serum antibodies against 2 proteinases were assayed using S. schenckii inoculated hairless mice. Subsequent to an intracutaneous injection of S. schenckii to the mouse skin, nodules spontaneously formed and disappeared for a period of 4 weeks. Histopathological examination results were in accordance with the microscopic observations. Micro-organisms disappeared during the fourth week. Serum antibody titers against purified proteinases I and II were measured weekly, using enzyme-linked immunosorbent assay (EIA). As a result, the time course of the antibody titers to both proteinases I and II were parallel to that of macroscopic and microscopic observations in an experimental mouse sporotrichosis model. These results suggest that S. schenckii produces both proteinases I and II in vivo. Moreover, the detection of antibodies against these proteinases can contribute to a serodiagnosis of sporotrichosis.

Effects of proteinase inhibitors on the cutaneous lesion of Sporothrix schenckii inoculated hairless mice.

Sporothrix schenckii produces two extracellular proteinases, namely proteinase I and II. Proteinase I is a serine proteinase, inhibited by chymostatin. On the other hand, proteinase II is an aspartic proteinase, inhibited by pepstatin. The addition of either pepstatin or chymostatin to the culture medium did not inhibit cell growth, however the addition of both inhibitors strongly inhibited fungal growth. Accordingly, this suggested that extracellular proteinases play an important role in cell growth and that such cell growth may be suppressed if these proteinases are inhibited. In order to substantiate this speculation in sporotrichosis, the effects of proteinase inhibitors on the cutaneous lesions of mice were studied. Ointments containing 0.1% chymostatin, 0.1% pepstatin and 0.1% chymostatin-0.1% pepstatin were applied twice daily on the inoculation sites of hairless mouse skin, and the time courses of the lesions examined. The inhibitory effect in vivo on S. schenckii was similar to that demonstrated in our previous in vitro study. Compared to the control, the time course curve of the number of nodules present after the application of either pepstatin or chymostatin was slightly suppressed. The application of both pepstatin and chymostatin, however, strongly suppressed nodule formation. This study not only confirmed the role of 2 proteinases of S, schenckii for fungal growth in vivo, but also may lead to their use as new topical therapeutic agents.

A speculative view of atopic dermatitis: barrier dysfunction in pathogenesis.

The development of atopy or atopic dermatitis is undoubtedly influenced by immunological abnormality. However, the question of factors which induce tolerance breakthrough and hypersensitivity to non-selective multiple antigens in atopy has remained unanswered. The broad clinical spectrum of this disease cannot be explained simply by allergy or immunological abnormalities. This speculative article is the first attempt to answer these questions and to explain the concept of atopic dermatitis by the barrier dysfunction theory based on our experimental results which demonstrated and increased permeability of the stratum corneum in patients with atopic dermatitis. We consider immunological abnormality and mucocutaneous barrier dysfunction as two major defects of atopy or atopic dermatitis. The wide range of clinical manifestations presented by this disease can be more easily comprehended by partial overlapping of these two core abnormalities. A mucocutaneous barrier defect readily allows penetration of multiple antigens or haptens. Repeated encounters with allergens may induce not only tolerance breakthrough and non-selective hypersensitivities but also enhancement of the allergic inflammation. On the other hand, an allergic inflammation stemming from the immunological abnormalities breaks down barrier functions. This sequence results in a 'vicious circle', which plays the most important role in pathogenesis of atopic dermatitis and probably of other atopic diseases.

Histopathological studies of Sporothrix schenckii-inoculated mice. Possible functions of polymorphonuclear leukocytes in normal and immunocompromised (congenitally athymic nude) mice.

Defense mechanisms against Sporothrix schenckii were studied using mouse models. After an intracutaneous injection of the yeast form of S. schenckii to the dorsal skin of the congenitally athymic nude and normal heterozygote littermate mice, nodules were formed. They regressed and disappeared in 10 weeks in the case of normal mice. On the other hand, nodules and then ulceration developed progressively in nude mice until all animals expired by dissemination of microorganisms at the 11th week of inoculation. Histopathologically the migrated cells were similar in both the normal and the nude mice, particularly during the early phase (within 24 h), with infiltration by PMNs being predominant. Fragmentation of S. schenckii commenced early during the 12-24 h stage of inoculation in the normal mice, while such fragmentation was scarce in nude mice even though numerous PMNs accumulated. Microscopic observations in the early stages (within 24 h of inoculation) suggested that the lack of killing activity by PMNs in nude mice contributes more to the impaired defense than the lack of macrophage activation by T-cells.

Skin barrier defect in atopic dermatitis: increased permeability of the stratum corneum using dimethyl sulfoxide and theophylline.

The existence of a defect in the skin barrier of patients with atopic dermatitis (AD) was demonstrated and its importance in the pathogenesis of AD was emphasized. In order to evaluate the penetration properties of the stratum corneum of AD patients, the in vivo skin response to the penetration of dimethyl sulfoxide (DMSO) and in vitro response to the penetration of theophylline utilizing a diffusion chamber were studied. Both methods demonstrated an increasing level of penetration through the epidermal stratum corneum, with greatest penetration being evident with lesional skin, followed by AD non-lesional and then the normal control. However, statistical significances existed only between non-lesional and lesional skins in the case of the DMSO test, and between the normal control and non-lesional skin in the case of the diffusion chamber analysis using theophylline. Increased penetration of a non-specific nature is important in the pathogenesis of AD.

A proposed guideline for psoralen photochemotherapy (PUVA) with atopic dermatitis: successful therapeutic effect on severe and intractable cases.

Psoralen photochemotherapy with UVA (PUVA) has been reported to be successfully substitutional for, or an adjunct to, conventional treatments in patients with atopic dermatitis (AD). Against the considerable advantages of utilizing PUVA for AD patients, however, it must also be balanced against the possible hazards for individual patients. We attempted herein to formulate a guideline for the selection of AD patients assigned to PUVA. According to this guideline, 114 patients were selected for PUVA treatment. Forty-five percent of the patients did not respond adequately to other conventional forms of treatment. Side effects from former treatments, particularly steroids, appeared in 39% of the patients. Subsequent to the treatments, the skin lesions significantly decreased in 81% of the inpatients and 67% of the outpatients, while some patient's lesions disappeared, despite that other forms of treatment had been unsuccessful in many cases.

Lipase of Malassezia furfur: some properties and their relationship to cell growth.

Lipase activity of Malassezia furfur, detected with alpha-naphthyl palmitate as a substrate, appeared to be associated with the insoluble fraction of the organism. Profiles of M. furfur lipase were similar to those of Candida cylindracea lipase. The pH optimum of the lipase was acidic, pH 5.0, which is very similar to skin surface pH. The enzyme activity was strongly activated by a lipase activator, sodium taurocholate (STC). Addition of STC to the culture medium activated cell growth in a dose-dependent manner and induced hyphae production. These results suggest that M. furfur lipase plays an important role in cell growth.