Five Japanese cases of antidesmoglein 1 antibody-positive and antidesmoglein 3 antibody-negative pemphigus with oral lesions.

BACKGROUND: Oral mucosal lesions develop in pemphigus vulgaris, but not in pemphigus foliaceus. This clinical phenomenon is explained by the 'desmoglein (Dsg) compensation theory'. Dsg3 and Dsg1 are major autoantigens for pemphigus vulgaris and pemphigus foliaceus, respectively. Dsg3 is overexpressed and Dsg1 is weakly expressed on the oral mucosa. Thus, on the oral mucosa, suppression of Dsg3 function by anti-Dsg3 autoantibodies is not compensated by weakly expressed Dsg1 in pemphigus vulgaris, while suppression of Dsg1 function by anti-Dsg1 autoantibodies is perfectly compensated by richly expressed Dsg3 in pemphigus foliaceus. OBJECTIVES: We present five Japanese patients with pemphigus who deviate from this theory, i.e. all patients showed oral lesions (three also had cutaneous lesions) and reacted only with Dsg1, but not with Dsg3, by enzyme-linked immunosorbent assay. METHODS: To confirm whether the unique clinical phenotypes in our patients were due to a different immunological profile from that in classical pemphigus, we examined the reactivity of the patient sera by immunoprecipitation-immunoblotting analysis using five Dsg1/Dsg2 domain-swapped molecules. RESULTS: The sera of two patients who had only oral lesions tended to react with the extracellular (EC) 5 domain of Dsg1, the domain that is considered nonpathogenic in classical pemphigus foliaceus. Sera of three patients with mucocutaneous lesions reacted with EC1 domain or with both EC1 and EC2 domains of Dsg1, like classical pemphigus foliaceus. CONCLUSIONS: These results indicate that antigenic diversity of anti-Dsg1 antibodies in these patients may cause the unique oral mucosal and cutaneous lesions, although further studies are required to elucidate the pathomechanisms.

Diabetic neuropathy is closely associated with arterial stiffening and thickness in Type 2 diabetes.

AIMS: Interaction of vascular and metabolic factors appears to contribute to the pathogenesis of diabetic neuropathy. The aim of the study was to assess the impact of arterial stiffening and thickness on diabetic neuropathy in Type 2 diabetes. METHODS: In 294 patients with Type 2 diabetes, neuropathy was assessed by four components: the presence of neuropathic symptoms, the absence of ankle tendon reflexes, perception of vibration scores and heart rate variation. We measured intima-media thickness (IMT) of carotid arteries to assess arterial thickening, and brachial-ankle pulse-wave velocity (PWV) and brachial pulse pressure (PP) which reflect arterial stiffening. RESULTS: Diabetic neuropathy, defined as > or = two of the four components, was significantly associated with age, duration, glycated haemoglobin (HbA(1c)), systolic blood pressure, diastolic blood pressure, PP, hypertension, retinopathy, urinary albumin excretion rate, nephropathy stages, PWV and IMT. PWV and PP were significantly associated with neuropathy independent of conventional cardiovascular risk factors. Multiple logistic regression analysis revealed that PWV, retinopathy, age, and HbA(1c), were significant independent determinants of neuropathy. CONCLUSIONS: The present cross-sectional study indicates that markers for vascular wall properties such as PWV, IMT and PP are significantly associated with diabetic neuropathy. PWV and PP are significant determinants of neuropathy independent of conventional cardiovascular risk factors. Multifactorial intervention to inhibit progression of the atherosclerotic process may slow progression of neuropathy.

Relationship between metabolic syndrome components and vascular properties in Japanese type 2 diabetic patients without cardiovascular disease or nephropathy.

To investigate the effect of metabolic syndrome (MS) components on early atherosclerosis markers, i.e., urinary albumin excretion rate (UAE), pulse wave velocity (PWV), and carotid intima-media thickness (IMT), we studied 536 Japanese patients with type 2 diabetes without cardiovascular disease or nephropathy. The MS definition by ATP III was employed. UAE, PWV, and IMT increased significantly with increasing the number of components even before fulfilling the diagnosis of MS. UAE was significantly influenced by high blood pressure, high triglycerides, and low HDL cholesterol. PWV was significantly increased by high blood pressure. IMT was significantly increased by high blood pressure and abdominal obesity. Multiple regression analysis, including MS components and putative risk factors, indicated that the number of MS components, age and glycosylated HbA1C were independent determinants of UAE, PWV, and IMT. LDL cholesterol and male gender were independent determinants of IMT. In conclusion, UAE, PWV, and IMT increased according to increasing the number of MS in type 2 diabetic patients without cardiovascular disease or diabetic nephropathy. The current observation considering the modifiable factors may help to identify patients who are at high risk of experiencing cardiovascular disease.

Thyroid gland tumour, pemphigus foliaceus and myasthenia gravis in the daughter of a woman with myasthenia gravis.

We describe a rare case of pemphigus foliaceus associated with familial myasthenia gravis (MG). A 35-year-old woman developed MG during oral corticosteroid treatment for pemphigus foliaceus. She had been operated on for a thyroid gland tumour that was confirmed histopathologically to be papillary carcinoma without metastasis. At the time of treatment, her mother had had MG for 30 years and undergone thymectomy 22 years ago. A specific ELISA technique showed that antidesmoglein 1 antibody was present in the daughter. There are many reports of multiple diseases such as pemphigus, thymoma, malignancy, and other autoimmune diseases associated with MG. However, familial MG following pemphigus foliaceus has not been reported previously.

Induction of C-reactive protein, serum amyloid P component, and kininogens in the submandibular and lacrimal glands of rats with experimentally induced inflammation.

The mRNAs for acute-phase proteins and kininogens were found to be increased in the submandibular gland (SMG) and extraorbital and intraorbital lacrimal gland (ELG and ILG) in response to experimentally induced inflammation in rats; i.e., 24 hours after subcutaneous injection of turpentine oil, mRNAs for C-reactive protein (CRP), serum amyloid P component (SAP), and H- and T-kininogens were induced in the SMG, ELG, and ILG of rats, whereas these mRNAs were not detected in the same tissues of normal control rats. The induction of mRNAs for these inflammatory proteins by turpentine oil was preceded by a transient increase in the level of mRNA for tumor necrosis factor-alpha (TNF-alpha) at 6 hours after subcutaneous injection of the oil. This was confirmed by injection of another inflammation inducer, lipopolysaccharide (LPS), which induced the TNF-alpha mRNA in the same way at 6 hours as turpentine oil did. The up-regulation of acute-phase proteins including kininogens in the SMG, ELG, and ILG suggest the existence of a strict defense system in the exocrine glands.

Allergic contact dermatitis from a pyridine derivative in polyvinyl chloride leather.

Antimicrobial coating of household products has gained wide acceptance in Japan in the past several years. Pyridine derivatives, used as antifungal or antibacterial agents in many common products, are known to cause contact dermatitis. We present a case of severe contact dermatitis caused by a pyridine derivative used as an antifungal agent in the polyvinyl chloride (PVC) leather of a chair. An open patch test was performed with each ingredient of the PVC leather. Other products were previously eliminated from consideration based on a series of negative patch tests. The PVC leather obtained from the patient's chair gave a ++ reaction with evident blistering, according to the International Contact Dermatitis Research Group standard. Fifteen ingredients of the PVC leather were open patch tested; a positive reaction was found with 2,3,5,6-tetrachloro-4 (methylsulphonyl) pyridine (1% in petrolatum). Clinicians should be aware that antifungal or antibacterial agents may be increasingly incorporated into common household products and should be suspected in cases of contact dermatitis.

Expression of kininogens in the connective tissue-type mast cells of the rat.

The connective tissue-type mast cells present in the submandibular gland (SMG) and peritoneal cavity of rats were found to express kininogens (KGs), the expression of which was demonstrated by Western blotting, reverse transcription-polymerase chain reaction (RT-PCR), RT-PCR Southern blotting, and light- and electron-microscopic immunocytochemistry. In the SMG, the analysis of cDNA amplified by RT-PCR revealed that the molecular species of mRNAs expressed were high-molecular-weight (HMW)-K KG and T-I KG. Light microscopic immunocytochemistry exclusively localized the KG protein(s) in the mast cells present in the SMG. The signals in the mast cells were very strong, but no positive reaction was observed in the granular convoluted tubular cells, acinar cells or striated duct cells. As determined by using electron microscopy, extremely strong labelling with immunogold was observed in the secretory granules of the mast cells, but no labelling in their nucleus or cytoplasm. Analysis by Western blotting and RT-PCR Southern blotting indicated that both protein and mRNA of KGs were present in the mast cells separated from the peritoneal cavity, indicating de novo synthesis of KG in these cells. Preliminary experiments implied that the connective tissue-type mast cells in other rat tissues also expressed KG.

Antimicrobial effects of acidic hot-spring water on Staphylococcus aureus strains isolated from atopic dermatitis patients.

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.

The production of superantigenic exotoxins by coagulase-negative staphylococci isolated from human skin lesions.

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).

Adherence characteristics and susceptibility to antimicrobial agents of Staphylococcus aureus strains isolated from skin infections and atopic dermatitis.

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.

Highly regulated expression of subtilisin-like proprotein convertase PACE4 (SPC4) during dentinogenesis.

Expressions of mRNAs for four subtilisin-like proprotein convertases (SPCs: furin, PACE4, PC6, and PC8) and bone morphogenetic protein 4 (BMP4) in the rat molar tooth during development were analyzed by Northern blotting, reverse transcriptase-polymerase chain reaction (RT-PCR), and in situ hybridization to explore the possible involvement of SPCs in the processing of proBMPs. We found a temporospacial expression of PACE4, but not one of the other SPCs, in this tissue; i.e., RT-PCR analysis revealed that the level of PACE4 mRNA, but not that of the other SPC mRNAs became high around the second postnatal day. This increase was in good accordance with the increase in BMP4 mRNA, indicating an apparent association of these molecules with the differentiation and establishment of functional ameloblasts and odontoblasts. During dentinogenesis, PACE4 mRNA was localized in the ameloblasts and odontoblasts. These observations suggest that PACE4 plays a crucial role in dentinogenesis, especially via the activation of BMPs.

High-resolution imaging of coronary calcifications by intense low-energy fluoroscopic X-ray obtained from synchrotron radiation.

PURPOSE: To obtain an intense monochromatic low-energy X-ray from synchrotron radiation (SR) and apply it to detect coronary calcifications. METHODS AND RESULTS: The SR beam was reflected with a silicon crystal to be expanded (150 mm in height and 80 mm in width) and to be monochromatized at an energy level of 37 keV. The X-ray was intermittently irradiated to obtain dynamic imaging of 30 images/s. Images were recorded by a digital fluorography system. The low-energy X-ray from SR sharply visualized calcification of coronary arteries, while conventional X-ray could not visualize coronary calcification. CONCLUSION: The intense monochromatic low-energy X-ray from SR is sensitive, has high-resolution for imaging coronary calcification and may serve as a screening method for coronary artery disease.

Involvement of vesicle-cytoskeleton interaction in AQP5 trafficking in AQP5-gene-transfected HSG cells.

A cDNA of rat aquaporin 5 (AQP5) was used to transfect to HSG (human salivary gland cells), and the trafficking mechanism was studied in vitro by confocal laser microscopy. The trafficking of AQP5 to the plasma membrane was induced by stimulation of AQP5-gene-transfected human salivary gland cells (HSGAQP5 cells) with thapsigargin, an inhibitor of endoplasmic Ca(2+)-ATPase, and or with A-23187, a calcium ionophore. Pretreatment of these cells with colchicine or vinblastine, microtubule inhibitors, prevented the trafficking induced by thapsigargin or A-23187. The trafficking event was not completely inhibited by cytochalasin B, a microfilament inhibitor. These results demonstrate that the trafficking of AQP5 vesicles to the plasma membrane is triggered by an increase in intracellular Ca(2+) and that the interaction of AQP5-containing vesicles with the cytoskeleton is involved in this trafficking.

Characteristics in adherence of streptococci and Staphylococcus aureus isolated from various infective skin lesions: serum IgA decreases adherence of Streptococcus pyogenes but not Staphylococcus aureus.

We characterized adherence of streptococci and Staphylococcus aureus strains isolated from various infective skin lesions in terms of hydrophobicity, negative charge, tube adherence, slime production, and influence on adherence to coverslips by plasma and serum immunoglobulins. High hydrophobicity was more frequently observed in Streptococcus pyogenes strains than in Streptococcus agalactiae strains (P < 0.01) and S. aureus strains (P < 0.001) and slime production was more frequently observed in S. agalactiae strains than in S. pyogenes strains (P < 0.05). Serum IgA decreased adherence to coverslips of S. pyogenes strains but not that of S. aureus strains.

Possible influences of Staphylococcus aureus on atopic dermatitis-- the colonizing features and the effects of staphylococcal enterotoxins.

BACKGROUND: Heavy colonization of atopic dermatitis (AD) with Staphylococcus aureus is well documented. This phenomenon suggests that S. aureus in AD lesions influences the disease processes of AD. OBJECTIVE: We describe the importance of the presence of S. aureus and staphylococcal enterotoxins A and B (SEA, SEB) in AD lesions. METHODS: We investigated the colonizing features of S. aureus in AD lesions using electron microscopy, the distribution of SEB in the eczematous skin of AD using immunofluorescence, the effects of SEA and SEB on normal human epidermal keratinocytes in organ culture, and the presence of specific IgE antibodies to SEA and/or SEB in serum of AD patients by enzyme immunoassay. RESULTS: S. aureus in AD lesions colonized on and in the horny layers of the eczematous skin. SEB produced by S. aureus was distributed mainly on the dermal-infiltrated cells, especially on eosinophils. SEA and SEB stimulated expression of ICAM-1 and HLA-DR in normal human keratinocytes. More than half of the AD patients in the present study had specific IgE antibodies to SEA and/or SEB in their serum. CONCLUSION: S. aureus and SEs have important roles in the exacerbation and prolongation of AD.

Dynamic intravenous coronary angiography using 2D monochromatic synchrotron radiation.

A method of examination for coronary artery disease that is less invasive and easier than coronary angiography (CAG) has been sought. We have developed a dynamic intravenous coronary angiography (IVCAG) system using synchrotron radiation (SR) and have used it clinically. Four patients suspected of having angina pectoris underwent IVCAG. An SR beam was reflected asymmetrically with a silicon crystal to produce a wide (150 mm x 80 mm) and monochromatic (37 keV) X-ray beam, with an energy level to achieve high sensitivity to the contrast agent. Following an intravenous injection of contrast agent, irradiation was applied for 4 ms periods at 33 ms intervals for dynamic IVCAG at 30 images s-1. Images were acquired with an image intensifier and recorded with a digital fluorography system. The dynamic images permitted clear visualization of the coronary arteries and permitted evaluation of coronary anatomy. Two patients exhibited no stenotic lesions, one patient had a 90% stenosis in the right coronary artery, and the remaining patient had a 25% stenosis at the site of previous percutaneous transluminal coronary angioplasty in the left anterior descending artery (LAD). The total irradiation doses used for IVCAG were less than those for conventional angiography. Dynamic IVCAG can be readily used for the evaluation of coronary arteries.

A common signaling pathway via Syk and Lyn tyrosine kinases generated from capping of the sialomucins CD34 and CD43 in immature hematopoietic cells.

The sialomucin CD34 is a useful marker for hematopoietic stem/progenitor cells. However, the role of CD34 remains poorly understood. Here we investigate the functions of CD34 and another sialomucin CD43 coexpressed on hematopoietic stem/progenitor cells. Stimulation of undifferentiated hematopoietic KG1a cells with anti-CD34 or anti-CD43 induced homotypic cytoadhesion, accompanied by formation of a long-lived cap of CD34 and CD43 respectively, which colocalized with F-actin. Stimulation with either antibody specifically increased tyrosine phosphorylation of the identical set of proteins of Lyn, Syk, pp60, pp69, and pp77 at the capping site. These events were similar to those observed in monocytic U937 cells ectopically expressing CD34. After stimulation of KG1a cells, coimmunoprecipitation of Lyn with pp69 and pp77 and of Syk with pp37 was detected in the membrane fraction. Blockade of antibody-induced cap formation by treatment with cytochalasin D leads to inhibition of tyrosine phosphorylation of Syk and pp77 and homotypic cytoadhesion. Moreover, normal human CD34(+) bone marrow cells showed cap formation of CD34 or CD43 after stimulation. These results suggest that crosslinking of either CD34 or CD43 activates the same signaling pathway for cytoadhesion through Lyn, Syk, and the novel tyrosine-phosphorylated proteins within hematopoiesis.