In situ strategy for biomedical target localization via nanogold nucleation and secondary growth.

Immunocytochemistry visualizes the exact spatial location of target molecules. The most common strategy for ultrastructural immunocytochemistry is the conjugation of nanogold particles to antibodies as probes. However, conventional nanogold labelling requires time-consuming nanogold probe preparation and ultrathin sectioning of cell/tissue samples. Here, we introduce an in situ strategy involving nanogold nucleation in immunoenzymatic products on universal paraffin/cryostat sections and provide unique insight into nanogold development under hot-humid air conditions. Nanogold particles were specifically localized on kidney podocytes to target synaptopodin. Transmission electron microscopy revealed secondary growth and self-assembly that could be experimentally controlled by bovine serum albumin stabilization and phosphate-buffered saline acceleration. Valuable retrospective nanogold labelling for gastric H+/K+-ATPase was achieved on vintage immunoenzymatic deposits after a long lapse of 15 years (i.e., 15-year-old deposits). The present in situ nanogold labelling is anticipated to fill the gap between light and electron microscopy to correlate cell/tissue structure and function.

Reversible lesion in the splenium of the corpus callosum in a patient with chronic alcoholism.

Marchiafava-Bignami disease (MBD) is often diagnosed in chronic alcoholics. The disease processes typically involve the corpus callosum and clinically presents with various manifestations on the basis of clinical condition, extent of the splenium of the corpus callosum involvement at brain magnetic resonance imaging (MRI), and prognosis. We report a patient affected by MBD, who presented an isolated reversible splenial lesion at brain MRI and achieved a favorable recovery.

Informative three-dimensional survey of cell/tissue architectures in thick paraffin sections by simple low-vacuum scanning electron microscopy.

Recent advances in bio-medical research, such as the production of regenerative organs from stem cells, require three-dimensional analysis of cell/tissue architectures. High-resolution imaging by electron microscopy is the best way to elucidate complex cell/tissue architectures, but the conventional method requires a skillful and time-consuming preparation. The present study developed a three-dimensional survey method for assessing cell/tissue architectures in 30-µm-thick paraffin sections by taking advantage of backscattered electron imaging in a low-vacuum scanning electron microscope. As a result, in the kidney, the podocytes and their processes were clearly observed to cover the glomerulus. The 30 µm thickness facilitated an investigation on face-side (instead of sectioned) images of the epithelium and endothelium, which are rarely seen within conventional thin sections. In the testis, differentiated spermatozoa were three-dimensionally assembled in the middle of the seminiferous tubule. Further application to vascular-injury thrombus formation revealed the distinctive networks of fibrin fibres and platelets, capturing the erythrocytes into the thrombus. The four-segmented BSE detector provided topographic bird's-eye images that allowed a three-dimensional understanding of the cell/tissue architectures at the electron-microscopic level. Here, we describe the precise procedures of this imaging method and provide representative electron micrographs of normal rat organs, experimental thrombus formation, and three-dimensionally cultured tumour cells.

Healthcare-associated infections in rheumatology in Japan.

Prospective observational study was performed to elucidate the incidence and characteristics of healthcare-associated infections in a university hospital for rheumatology care. In this study, a total of 1,226 patients were prospectively enrolled between March 2004 and February 2006 and between April 2008 and December 2008. Healthcare-associated infection was defined as an infection developing after the third day of admission to the rheumatology ward. We detected the following 54 healthcare-associated infections in 49 patients: respiratory tract infection, 14 cases; Clostridium difficile infection, 2 cases; urinary tract infection, 4 cases; bloodstream infection, 9 cases; skin infection, 2 cases; reactivation of latent cytomegalovirus infection, 6 cases; herpes zoster infection, 5 cases; Candida infection, 7 cases; others, 4 cases. The incidence rate of respiratory tract infection was the highest. Methicillin-resistant Staphylococcus aureus was the causative bacterium in 21% of respiratory tract infections cases. Bloodstream infection due to the insertion of a catheter and opportunistic infection by a latent virus were also occurred commonly. Respiratory tract infection, bloodstream infection and opportunistic infection by a latent virus were the most common causes of healthcare-associated infection in rheumatology. It is important to pay more attention to healthcare-associated infection.

A scale-free structure prior for graphical models with applications in functional genomics.

The problem of reconstructing large-scale, gene regulatory networks from gene expression data has garnered considerable attention in bioinformatics over the past decade with the graphical modeling paradigm having emerged as a popular framework for inference. Analysis in a full Bayesian setting is contingent upon the assignment of a so-called structure prior-a probability distribution on networks, encoding a priori biological knowledge either in the form of supplemental data or high-level topological features. A key topological consideration is that a wide range of cellular networks are approximately scale-free, meaning that the fraction, , of nodes in a network with degree is roughly described by a power-law with exponent between and . The standard practice, however, is to utilize a random structure prior, which favors networks with binomially distributed degree distributions. In this paper, we introduce a scale-free structure prior for graphical models based on the formula for the probability of a network under a simple scale-free network model. Unlike the random structure prior, its scale-free counterpart requires a node labeling as a parameter. In order to use this prior for large-scale network inference, we design a novel Metropolis-Hastings sampler for graphical models that includes a node labeling as a state space variable. In a simulation study, we demonstrate that the scale-free structure prior outperforms the random structure prior at recovering scale-free networks while at the same time retains the ability to recover random networks. We then estimate a gene association network from gene expression data taken from a breast cancer tumor study, showing that scale-free structure prior recovers hubs, including the previously unknown hub SLC39A6, which is a zinc transporter that has been implicated with the spread of breast cancer to the lymph nodes. Our analysis of the breast cancer expression data underscores the value of the scale-free structure prior as an instrument to aid in the identification of candidate hub genes with the potential to direct the hypotheses of molecular biologists, and thus drive future experiments.

Resolution of anti-neutrophil cytoplasmic antibody-associated vasculitis after resection of gastric cancer.

We report a case of anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis in a 62-year-old patient with gastric cancer. The myeloperoxidase-anti-neutrophil cytoplasmic antibody (MPO-ANCA) level was threefold above normal preoperatively. Vasculitis was seen on renal biopsy. Gastric resection revealed well-differentiated adenocarcinoma and vasculitis. The MPO-ANCA level returned to normal post-operatively. Although ANCA-associated vasculitis occasionally accompanies malignant tumors, this is the first documented case of concurrent gastric cancer-associated and ANCA-associated vasculitis, with post-operative resolution of the vasculitis.

Protein-losing gastroenteropathy associated with primary Sjögren's syndrome: a characteristic oriental variant.

Protein-losing gastroenteropathy (PLGE) is a rare manifestation of primary Sjögren's syndrome (SS). We report a case of a 41-year-old Japanese man, who is the first male patient, with PLGE associated with primary SS. Although serum anti-SSA and SSB antibodies were detected, he had no subjective sicca symptoms. He had multiple annular erythema: a characteristic skin manifestation of Asian SS patients. A diagnosis of PLGE was made from results of (99m)Tc-labelled albumin scintigraphy and a faecal alpha-1-antitrypsin clearance test. Intravenous administration of high-dose glucocorticoid was not effective, but pulse methylprednisolone therapy alleviated disease manifestations. As all cases of PLGE associated with primary SS have been reported from East Asia, this complication could be essentially limited to Asian patients.

Sjögren's syndrome associated with chronic hepatitis C, severe thrombocytopenia, hypertrophic cardiomyopathy, and diabetes mellitus.

A 74-year-old woman with Sjögren's syndrome and chronic hepatitis C (CHC) was admitted to our hospital in October 2003 for treatment of diabetes mellitus. She had the past history of recurrent thrombocytopenia, which was proven to be due to peripheral destruction. Although she had been diagnosed with hypertrophic cardiomyopathy (HCM) for 2 years, she had never felt palpitation. She suddenly died probably of fatal arrhythmia related to HCM during the last hospitalization. Although hepatitis C virus (HCV) infection has been associated with Sjögren's syndrome, thrombocytopenia, HCM, and diabetes mellitus, all these diseases rarely occur in a single patient. It will be necessary to identify similar cases to elucidate the etiopathogenesis of extra-hepatic manifestations of HCV infection.

Muscular polyarteritis nodosa as a cause of fever of undetermined origin: a case report and review of the literature.

We describe an unusual presentation of a localized form of polyarteritis nodosa (PAN) manifested by fever of undetermined origin (FUO). Biopsies of the gastrocnemius muscle revealed necrotizing arteritis and initiation of prednisolone (PSL) brought rapid response. The PAN localized to muscle is rare; furthermore, this disease presented as FUO is very rare. We want to increase awareness that muscle can be also a single-affected site as well as other well known sites such as appendix, gallbladder, uterus or testis, and skin. Since there is no single appellation for this disease, we would like to propose the term "muscular PAN".

Kikuchi's disease involving the supraclavicular lymph nodes and associated with transient eruption.

A 26-year-old woman presented with high fever, marked supraclavicular lymphadenopathy, and morbilliform eruptions and was diagnosed with Kikuchi's disease (KD) based on pathologic findings from biopsied lymph nodes. All her manifestations of KD improved, however, without any specific treatment. The picture of transient morbilliform eruptions typified in KD here is seldom shown in the literature. In general, KD would run a benign course of supraclavicular lymphadenopathy.

Rheumatoid arthritis complicated with acute interstitial pneumonia induced by leflunomide as an adverse reaction.

A 49-year-old Japanese man with rheumatoid arthritis acutely developed a skin eruption and severe non-productive cough seventeen days after the administration of leflunomide. Because all bacteriology findings were negative, steroid pulse-therapy was initiated promptly due to the rapidity of chest X-ray progression and the deterioration of arterial blood oxygen pressure. Although cough was induced by methotrexate, interstitial pneumonia was not detected clinically before leflunomide administration. He finally died of respiratory failure 128 days after the onset of acute interstitial pneumonia. According to the post-market surveillance, as high as approximately 1.1% of the patients on,leflunomide have developed interstitial pneumonia in Japan. It is important to emphasize that acute interstitial pneumonia due to leflunomide is a very severe and potentially fatal side effect.

[Systemic lupus erythematosus with steroid induced non-alcoholic steatohepatitis: a case report].

A 56-year old Japanese female was admitted to our hospital because of the increased levels of serum AST, ALT, and gamma-GTP. She was diagnosed with systemic lupus erythematosus in September, 1996 and had been on a regular glucocorticoid therapy since then. Abdominal ultrasonography showed the mild fatty liver, and hepatic histopathology revealed a typical and remarkable steatohepatitis, a remarkable neutrophil infiltration, and Mallory bodies. Because she had no history of alcohol-drinking, diagnosis of non-alcoholic steatohepatitis (NASH) was made. Treatment was started with a low-calorie diet, bed-rest, and an oral administration of alpha-tocopherol and bezafibrate with favorable effects on her serum levels of AST, ALT, gamma-GTP, and LDH. When a patient on a glucocorticoid therapy shows signs of fatty liver, diabetes mellitus, hyperlipidemia, an insulin resistance, NASH should be considered as one of the differential diagnosis. This is particularly important since proper therapy with a low-calorie diet and drugs with anti-oxidant activities improve this potentially progressive disease before resulting in liver cirrhosis and hepatic carcinoma.

Effects of FTY720 in MRL-lpr/lpr mice: therapeutic potential in systemic lupus erythematosus.

UNLABELLED: OBJECTIVE To examine the effects of a novel immunosuppressant, FTY720, on hematolymphoid cells and the clinical course of MRL-lpr/lpr (MRL/lpr) mice genetically predisposed to systemic lupus erythematosus (SLE). METHODS: Apoptosis of hematolymphoid cells was determined in vitro by FACScan after staining with propidium iodide or merocyanine 540. From 4 months of age, 15 female MRL/lpr mice received oral administration of 2 mg/kg each of FTY720, methylprednisolone (mPSL), or vehicle, 3 times per week. Therapeutic efficacy was evaluated by levels of anti-dsDNA antibodies in serum and the survival rate. In parallel, T cell proliferation and secretion of interleukin 2 (IL-2) induced by anti-CD3, phenotypes of the spleen, lymph node and bone marrow cells, as well as immunohistochemistry of the kidney, were examined in vitro. RESULTS: FTY720 at 2 microM induced apoptosis in more than 70% of double negative (CD4-/CD8-) T cells from the spleen of MRL/lpr mice in vitro. Oral FTY720 was tolerated well with no apparent side effects. FTY720 treated and control mice gained weight at an identical pace through to 9 months of age. FTY720 significantly suppressed the production of anti-dsDNA antibodies (FTY720 vs control: 1739 +/- 898 U/ml vs 410 +/- 356 U/ml at 8 months of age; p < 0.05) and reduced the deposition of IgG in glomeruli compared to control animals. At 9 months of age, the survival rate in the FTY720 treated mice was 86.9% compared to 33.0% in controls (p < 0.01). FTY720 decreased the number of double negative T cells from the spleen and lymph nodes in vivo, and increased T cell proliferation and IL-2 secretion induced by anti-CD3 stimulation in vitro. CONCLUSION: FTY720 suppressed the development of autoimmunity and prolonged the lifespan of female MRL/lpr mice. Suppression of autoimmunity, at least in part, may have resulted from an apoptogenic potential of FTY720. Hence, it could be useful for primary or adjunctive therapy of human SLE.