Raised serum levels of interleukins 6 and 8 and antiphospholipid antibodies in an adult patient with Henoch-Schönlein purpura.

BACKGROUND: The long-term prognosis of Henoch-Schönlein purpura (HSP) is determined by the severity of renal involvement, known as HSP nephritis, which varies considerably from patient to patient. There is now increasing evidence that dysregulated cytokine production plays a crucial role in human autoimmune and inflammatory processes. AIM: To explore the possible contributions of serum antistreptolysin O, C-reactive protein, IgA, interleukin (IL)-6, IL-8, tumour necrosis factor (TNF)-α, anticardiolipin antibody (aCL) and antiphosphatidylserine-prothrombin complex antibody (anti-PSPT) in the pathogenesis of HSP, and to evaluate correlations between those biological parameters and the clinical features. METHODS: Records were reviewed of 58 patients with HSP who presented initially with palpable purpura between 2003 and 2009. Serum IL-6 levels were determined by chemiluminescent enzyme immunoassay, IL-8 levels by ELISA and TNF-α levels by quantitative sandwich enzyme immunoassay. Serum aCL and anti-PSPT levels were measured according to our previously published procedures. RESULTS: There was a significant correlation between the serum IL-6 and IgA anti-PSPT levels, and also between the serum IL-8 and IgA anti-PSPT levels. Serum IL-8 and IgA aCL levels were both significantly higher in patients with renal involvement than in those without. Serum IL-6 and IgM anti-PSPT levels were also significantly higher in patients with gastrointestinal symptoms than in those without. CONCLUSIONS: We suggest that serum IL-6 and IL-8 associated with antiphospholipid antibodies play a pivotal role in the induction of HSP. Based on our results, IL-8 and IgA aCL levels could be useful as markers to monitor the development of HSP nephritis, and IL-6 and IgM anti-PSPT levels could be used as markers to monitor the development of gastrointestinal involvement.

IgA pemphigus with paraneoplastic pemphigus-like clinical features showing IgA antibodies to desmoglein 1/3 and desmocollin 3, and IgG and IgA antibodies to the basement membrane zone.

A 79-year-old Japanese woman presented with severe recalcitrant erosions on her oral mucosa, resembling paraneoplastic pemphigus. Using indirect immunofluorescence, we detected IgA antibodies against the cell surface, and both IgG and IgA antibodies against the basement membrane zone. Immunoblotting showed that the IgG antibodies reacted weakly with bullous pemphigoid 230 and periplakin, whereas the IgA antibodies did not react with any antigen. IgA antibodies to both desmoglein (Dsg)1 and Dsg3 were detected by ELISA. IgA antibodies to desmocollin (Dsc)3 were also detected by using cDNAs for human Dsc1-3 transfected into COS-7 cells. Despite treatment with oral prednisolone, high-dose intravenous immunoglobulin and double-filtration plasmapheresis, the skin lesions remained active, and the patient died from bronchiolitis obliterans-like respiratory failure. Despite extensive investigations and postmortem examination, no underlying neoplasms were found. The complex immunopathological findings probably played an important role in the development of the patient's unusual clinical features.

Elevated antilysosomal-associated membrane protein-2 antibody levels in patients with adult Henoch-Schönlein purpura.

BACKGROUND: Henoch-Schönlein purpura (HSP) is characterized by IgA-containing immune complexes within leucocytoclastic vasculitis. Lysosomal-associated membrane protein-2 (LAMP-2) was first identified as part of a systematic search for antineutrophil cytoplasmic antibody (ANCA) antigens expressed on neutrophils and endothelial cells. OBJECTIVES: To investigate the presence of ANCA in patients with adult HSP and microscopic polyangiitis (MPA), and to measure serum LAMP-2 antibody levels in these patients. METHODS: Twenty-four adult patients with HSP, eight with MPA and 24 normal healthy controls were examined. ANCA detection was performed using indirect immunofluorescence (IIF), a direct enzyme-linked immunosorbent assay (ELISA) and a capture ELISA specific for myeloperoxidase (MPO) and proteinase 3 (PR3). We measured other ANCA-associated antibodies including anti-LAMP-2 antibody in serum using ELISA. Immunohistochemical (IHC) staining was used for anti-LAMP-2 antibody expression in patient skin biopsies. To determine the cut-off value of the serum anti-LAMP-2 antibody, a receiver operating characteristic (ROC) curve was constructed using statistical analysis software (JMP 8·0·2; SAS Institute Inc., Cary, NC, U.S.A.). RESULTS: The sera of all patients with HSP were negative for MPO-ANCA and PR3-ANCA by direct ELISA and by capture ELISA. However, ANCA was present in 17 (71%) of the 24 patients with HSP based on IIF. In contrast, we found MPO-ANCA in all eight patients with MPA using both ELISA methods. We found serum anti-LAMP-2 antibody levels in HSP significantly higher than in MPA and in healthy individuals (P = 0·002 and P = 0·00167, respectively). The area under the curve of serum anti-LAMP-2 antibody between HSP and MPA was 0·8698 by ROC analysis. The optimal cut-off point was 0·267 U mL(-1) (sensitivity 1·000, specificity 0·583). We found a significant positive correlation between serum anti-LAMP-2 antibody levels and serum IgA levels in HSP (r(s) = 0·731, P = 0·00226). Anti-LAMP-2 antibody overexpression in IHC staining was present in 20 (83%) of the patients with HSP. The overexpression was observed within the neutrophils and endothelial cells of leucocytoclastic vasculitis. There was a significant positive correlation between IHC staining score and positive serum anti-LAMP-2 antibody. The 24 patients with HSP and the eight patients with MPA were negative for antiazurocidin antibodies, antibactericidal permeability increasing protein antibodies, anticathepsin G antibodies, antielastase antibodies, antilactoferrin antibodies and antilysozyme antibodies. CONCLUSIONS: We suggest that anti-LAMP-2 antibody could play some role in the pathogenesis of adult HSP, and have excluded a role for MPO-ANCA and PR3-ANCA. We propose that measuring serum anti-LAMP-2 antibody could be a feasible method of differential diagnosis between HSP and MPA.

Genome-wide mapping for fatty acid composition and melting point of fat in a purebred Duroc pig population.

The fatty acid composition and melting point of fatty tissue are among the most important economic traits in pig breeding because of their influence on the eating quality of meat. Identifying the quantitative trait locus (QTL) of these traits may help reveal the genetic structure of fatty acid composition and the melting point of fatty tissue and improve meat-quality traits by marker-assisted selection. We conducted whole-genome QTL analysis for fatty acid composition and melting point of inner and outer subcutaneous fat and inter- and intramuscular fat in a purebred Duroc population. A total of 129 markers were genotyped and used for QTL analysis. For fatty acid compositions of inner and outer subcutaneous fat, three significant QTL and 17 suggestive QTL were detected on SSC2, 4, 6, 8, 9, 10, 11, 12, 14 and 18. For the melting point of inner and outer subcutaneous fat, two significant QTL were detected on the same region of SSC14. For fatty acid compositions of inter- and intramuscular fat, five significant QTL and 13 suggestive QTL were detected on SSC2, 4, 6, 8, 9, 10, 14 and 15. On SSC14, significant QTL for C18:0 and C18:1 of outer subcutaneous fat and intramuscular fat, and melting point of subcutaneous fat, which had high likelihood of odds (LOD) scores (2.67-5.78), were detected in the same region. This study determined QTL affecting fatty acid composition and melting point of different fat tissues in purebred Duroc pigs.

Involvement of brown adipose tissue in subcutaneous fat necrosis of the newborn.

BACKGROUND: Subcutaneous fat necrosis (SCFN) of the newborn is a rare condition that manifests within days after birth. The interscapular region, axillae and shoulders are the most commonly affected sites, corresponding to anatomic sites of brown adipose tissue (BAT) in newborns. OBJECTIVE: We postulated a specific involvement of BAT in SCFN and searched for brown adipocytes at affected sites. METHODS: Biopsy specimens were immunostained with antibodies against uncoupling protein 1 (UCP-1) and examined by electron microscopy. We also examined BAT by (18)F-fluorodeoxyglucose ((18)F-FDG) positron emission tomography and computed tomography (PET-CT) scanning. RESULTS: A few cells in biopsy specimens from two patients bound antibodies against UCP-1, and brown adipocytes were detected at several stages of degeneration. PET-CT scans revealed lower uptake of (18)F-FDG at major sites of SCFN. CONCLUSION: SCFN and BAT can be found at the same sites, suggesting a pathophysiological connection.

Attitudes of dermatologists toward the Chapel Hill Consensus Conference nomenclature and classification.

This study was designed to explore the attitudes of dermatologists towards the Chapel Hill Consensus Conference (CHCC) nomenclature and classification. We developed a questionnaire to determine the views of chief assistant dermatological professors at 61 Japanese university hospitals. A chi2 analysis of the responses found a close relationship between dermatological facilities that based their evaluations on the CHCC and their likelihood of taking confirmatory skin biopsies from patients with suspected microscopic polyangiitis with cutaneous features. In those facilities, the physicians and pathologists tended to consider cutaneous polyarteritis nodosa and cutaneous leucocytoclastic angiitis as independent disease conditions. We believe that it would be beneficial for dermatologists to take advantage of the CHCC, through which an appropriate early diagnosis of vasculitis can be realized. The present investigation provides a picture of current practices of Japanese dermatologists with reference to the management of vasculitis, including the extent to which biopsies are used to establish the diagnosis.

Histopathological evidence of small-vessel vasculitis within the skin and lungs associated with interstitial pneumonia in an adult patient with dermatomyositis.

Dermatomyositis (DM) often has a poor prognosis, due to complications associated with malignancy or interstitial pneumonia (IP). It is uncommon to find histopathological small-vessel vasculitis within cutaneous lesions and pulmonary capillaritis in patients with DM. A 64-year-old woman was diagnosed with DM based on the presence of quadriparesis, typical heliotropic rash, Gottron's papules, increased serum levels of muscle enzymes and typical muscle biopsy findings. She also had associated IP, which subsequently developed into a rapidly progressive condition. We found a high titre of Krebs von den Lunge (KL)-6 in her serum. Measurement of serum KL-6 level is widely accepted as a diagnostic test to monitor the activity of interstitial lung diseases. Histopathological examinations of the cutaneous and lung-associated features showed small-vessel vasculitis. We suggest that investigation of KL-6 levels in patients with DM and associated conditions should be carried out to determine if raised KL-6 levels are useful at predicting outcome or severity of various features.

Enhanced skin regeneration by nanoegg formulation of all-trans retinoic acid.

All-trans retinoic acid (atRA) which could smooth wrinkles and produce less pigmented skin after a few months of treatment has been studied in research into topical treatments for a potent inhibitor of new melanin production. However, the clinical responses of commercial atRA cream predominantly comprise severe inflammation. We report a novel nanotechnology "nanoegg" system giving improved effects of atRA self-assembly which were coated by CaCO3. Dorsal areas of hairless mouse and porcine skin were employed for administration of nanoegg ointment and commercial products. The mRNA for heparin-binding epidermal growth factor-like growth factor (HB-EGF) from tissues was measured by a real-time PCR method. All tissues were stained for detection of hyaluronate and the thickness of the epidermis. A clinical trial in humans was carried out at St. Marianna University in Japan. As a result, the irritation and inflammation associated with atRA molecules were substantially reduced. The physicochemical instability of atRA was also dramatically improved. Furthermore, nanoegg enhanced marked expression of mRNA for HB-EGF from keratinocytes, which is known as one of the markers of keratinocyte turnover. Also, production of hyaluronate was surprisingly in the intercellular spaces of the basal and spinous cell layers 2 days after treatment. Even at the low concentration of atRA in the nanoegg system, the proliferation and differentiation of keratinocyte was somewhat enhanced. A nanoegg may thus not only prevent adverse effects, but also markedly enhance the main effect.

Immunohistochemical detection of the placental form of glutathione S-transferase in dysplastic and neoplastic human uterine cervix lesions.

Expression of the human placental form of glutathione S-transferase (GST-pi) in dysplasia (53 cases), carcinoma in situ (10 cases), and invasive carcinoma (46 cases) of human uterine cervix was investigated immunohistochemically with specific anti-GST-pi rabbit antibody. While normal squamous epithelium was largely negative, the binding of antibody was appreciable in mild and moderate dysplasias, especially in the cytoplasm of cells demonstrating koilocytotic atypia. In severe dysplasia, the nuclei as well as the cytoplasm were strongly stained in all cell layers except for the superficial layer, and in carcinoma in situ both of them were also strongly stained in all cell layers. In invasive carcinoma, over 90% of cases exhibited strong cytoplasmic staining and in over 70% the nuclei were positive. GST activity towards 1-chloro-2,4-dinitrobenzene and GST-pi protein content were significantly increased in all of 4 squamous cell carcinomas examined as compared to values for normal cervical epithelia. Two-dimensional gel electrophoresis followed by immunoblotting using the GST-pi antibody demonstrated that, of many cytoplasmic proteins, only the GST-pi subunit was specifically bound. These results indicate that GST-pi is a potentially useful immunohistochemical marker for (pre)neoplasia of human uterine cervix. In addition, it was demonstrated that the cells in severe dysplasia, carcinoma in situ, and invasive carcinoma expressing GST-pi were often characterized by staining with a monoclonal antibody to the v-H-ras gene product.

Purification and subunit-structural and immunological characterization of five glutathione S-transferases in human liver, and the acidic form as a hepatic tumor marker.

Five glutathione S-transferase (GST, EC 2.5.1.18) forms were purified from human liver by S-hexylglutathione affinity chromatography followed by chromatofocusing, and their subunit structures and immunological relationships to rat liver glutathione S-transferase forms were investigated. They were tentatively named GSTs I, II, III, IV and V in order of decreasing apparent isoelectric points (pI) on chromatofocusing. Their subunit molecular weights assessed on SDS-polyacrylamide gel electrophoresis were 27 (Mr X 10(-3)), 27, 27.7,27 and 26, respectively, (26, 26, 27, 26, and 24.5 on the assumption of rat GST subunit Ya, Yb and Yc as 25, 26.5 and 28, respectively), indicating that all forms are composed of two subunits identical in size. However, it was suggested by gel-isoelectric focusing in the presence of urea that GSTs I and IV are different homodimers, consisting of Y1 and Y4 subunits, respectively, which are of identical Mr but different pI, while GST II is a heterodimer composed of Y1 and Y4 subunits. This was confirmed by subunit recombination after guanidine hydrochloride treatment. GST III seemed to be identical with GST-mu with regard to Mr and pI. GST V was immunologically identical with the placental GST-pi. On double immunodiffusion or Western blotting using specific antibodies to rat glutathione S-transferases, GST I, II and IV were related to rat GST 1-1 (ligandin), GST III(mu) to rat GST 4-4 (D), and GST V (pi) to rat GST 7-7 (P), respectively. GST V (pi) was increased in hepatic tumors.

Chemical reactions of organic compounds on clay surfaces.

Chemical reactions of organic compounds including pesticides at the interlayer and exterior surfaces of clay minerals and with soil organic matter are reviewed. Representative reactions under moderate conditions possibly occurring in natural soils are described. Attempts have been made to clarify the importance of the chemical nature of molecules, their structures and their functional groups, and the Brönsted or Lewis acidity of clay minerals.

Survival benefit for patients with colorectal cancer detected by population-based screening program using an immunochemical fecal occult blood test.

The effectiveness of screening for colorectal cancer by an immunochemical fecal occult blood test was investigated by comparing the survival time of 194 screen-detected colorectal cancer subjects (screen-detected group) with that of 352 routinely diagnosed subjects (control group). The rate of Dukes' A cancers was significantly higher in screen-detected group than in the control group (43.8% vs 16.8%, p<0.0001), whereas that of Dukes' D cancer was the opposite (5.7% vs 21.3%, p<0.0001). The survival rate for screen-detected group was higher than that for the control group (Kaplan-Meier method, p<0.0001). By a proportional hazard model with adjustment for prognostic factors except stage, the risk ratio of death from colorectal cancer was 0.40 (p=0.0007) for screen-detected group versus the control group. Therefore, the screening was suggested to be effective in survival benefit, although there might be potential biases inherent in observational studies.

Surgical treatment of arch-type dissecting aortic aneurysms.

We have proposed calling dissecting aortic aneurysm with entry of the dissection in the aortic arch "arch-type". We described the procedures and results of surgery for 12 cases of this disorder. The dissection was localized (type a) in six cases, and extended (type b) in the other six. Seven cases were treated by patch or direct closure of the entry, with one operative death and two later deaths. Five cases were treated by reconstruction of the ascending aorta and the aortic arch with a prosthetic graft, with two operative deaths and one later death. Efforts must be directed towards improving the outcome of the latter operation.

Two cases of postoperative erythroderma.

Postoperative erythroderma is a rare disease of which the clinical course is almost always fatal. It is characterized by fever, skin rash, hepatic dysfunction and leukopenia, and many of the cases occur after cardiovascular surgery. We have experienced two cases of this disease which are suspected to be related to a large amount of blood transfusion. In the second case, we observed genetical mismatch of the HLA typing between the patient and his family members. This observation strongly suggests the possibility that postoperative erythroderma is a form of graft-versus-host (GVH) reaction induced by transfused lymphocytes. The reason why the patient's immunity is so suppressed as to cause GVH reaction is still unknown. Cardiopulmonary bypass and a large amount of fresh blood transfusion are suspected for immune suppression. The fact that postoperative erythroderma occurs almost exclusively in the Japanese cannot be fairly explained, either. Racial homogeneity in Japan might cause immune tolerance in blood transfusion.

A clinical study of cerebral circulation during extracorporeal circulation.

The objective of this study is to clarify the relationship of cerebral blood flow to extracorporeal circulation flow and mean arterial pressure during nonpulsatile extracorporeal circulation under moderate hypothermia. Cerebral blood flow was determined by an argon saturation and desaturation method after that of Pevsner and colleagues with a mass spectrometer in 21 adult patients undergoing cardiac operations. Cerebral blood flow was 25, 33, 35, and 42 ml/100 gm/min, ranging from 19 to 50 ml/100 gm/min, at extracorporeal circulation flow rates of 40, 50, 60, and 70 ml/kg/min, respectively. Cerebral blood flow increased proportionally to extracorporeal circulation flow. Cerebral blood flow scattered almost transversely to mean arterial pressure and was 31 ml/100 gm/min in a hypotensive group (mean arterial pressure 34 to 50 mm Hg) and 34 ml/100 gm/min in another group (mean arterial pressure 51 to 94 mm Hg). Mean arterial pressure did not significantly influence cerebral blood flow. Cerebral oxygen consumption did not remarkably decrease and remained in the reasonable range when cerebral blood flow was 23 to 40 ml/100 gm/min. Subsequently, we assumed that the average cerebral blood flow value of 25 ml/100 gm/min at an extracorporeal circulation flow rate of 40 ml/kg/min also would be in the safe range. All of the patients are living without cerebral complications. We conclude that (1) cerebral blood flow was extracorporeal circulation flow dependent and (2) cerebral blood flow in the safe range was maintained even in the hypotensive range, provided the extracorporeal circulation flow rate was 40 ml/kg/min or higher.

Enhanced expression of the high-affinity receptor for IgE (Fc(epsilon)RI) associated with decreased numbers of Langerhans cells in the lesional epidermis of atopic dermatitis.

Epidermal Langerhans cells (LCs) and the high-affinity receptor for IgE (Fc(epsilon)RI) on their surface are considered important in the pathogenesis of atopic dermatitis (AD). We investigated the numbers of epidermal LCs and their Fc(epsilon)RI expression in patients with AD and healthy controls. Biopsy specimens taken from lesional skin from 17 patients with AD, non-lesional skin from five patients with AD and normal skin from five healthy individuals were immunohistochemically stained with a monoclonal antibody against CD1a or with either of two monoclonal antibodies against two different epitopes of Fc(epsilon)RI alpha chain. Many dendritic cells were positively stained with anti-CD1a antibody in the epidermis of each skin sample, and fewer cells were stained with anti-Fc(epsilon)RI antibodies. The numbers of epidermal LCs positive for Fc(epsilon)RI were significantly increased in both lesional and non-lesional skin from AD patients compared with those in normal skin, suggesting important roles of Fc(epsilon)RI+LCs in the pathogenesis of the disease. In contrast, the numbers of total epidermal LCs (CD1a-positive) were decreased in AD lesional skin compared with those in non-lesional skin from AD patients and in normal skin from healthy subjects. Together with our finding that the numbers of epidermal LCs were negatively correlated with the clinical severity of the AD lesions, we concluded that epidermal LCs may decrease in some conditions of AD, probably in lesions with severe inflammation.

Clinical results of cardiopulmonary bypass with selective cerebral perfusion for aneurysm of the ascending aorta and the aortic arch.

The purpose of this report is to describe our experience using cardiopulmonary bypass with selective cerebral perfusion in 10 patients whose aneurysms required cross-clamping the aorta and brachiocephalic vessels. Cerebral perfusion was carried out with individual roller pumps. Flow rate was 11.2 +/- 4.8 ml per kilogram of body weight per minute to the right axillary artery, 8.5 +/- 1.0 to the right common carotid artery, 7.0 +/- 2.0 to the left common carotid artery, and 3.9 to the left subclavian artery. All the patients came off bypass smoothly, and only 3 required small amounts of inotropic agents postoperatively. One patient with mycotic aneurysm died in the fourth postoperative week of massive bleeding due to disruption of the suture line. No cerebral complications were experienced. The operative results of this series suggest the usefulness of cardiopulmonary bypass with selective cerebral perfusion, and reasonably favorable outlook for patients with aneurysm involving the aortic arch and the ascending aorta.

Transcortical sensory aphasia following left frontal infarction.

Two right-handed patients who exhibited language disability after left frontal infarction are described. The patients spoke fluently and exhibited excellent repetition ability from the onset of infarction without exhibiting any oral apraxia, but had deficits in auditory comprehension, naming, reading and writing. In both patients, brain magnetic resonance imaging (MRI) revealed infarction in the left inferior frontal gyrus, the middle frontal gyrus and the anterior part of the lower precentral gyrus. Single photon emission computed tomography (SPECT) revealed decreased blood flow in the same regions as those shown to be infarcted by MRI. The MRI and SPECT findings and the symptoms of these patients suggest that left frontal lesions that encompass Broca's area produce fluent aphasia if the posterior part of the left precentral gyrus or motor cortex remains intact and that lesions anterior to Broca's area and the middle frontal gyrus produce a deficit in auditory comprehension of single words as well as sentences.

Systemic lupus erythematosus associated with antiribosomal P protein antibody.

Three patients with systemic lupus erythematosus had positive antiribosomal P protein antibodies. The patients are women who had mild diseases that did not involve prominent internal organs. By using an immunofluorescence method for the detection fo antinuclear antibodies that employs HEp-2 cells, these serum samples produced intense stainings on cytoplasm and nucleoli that were digested with RNase. With the use of an immunoblotting technique, these serum samples recognized three ribosomal P proteins, called PO (38 kd), P1 (19 kd), and P2 (17 kd). We suggest that antiribosomal P protein antibodies can be detected in patients with systemic lupus erythematosus and associated mild clinical features.