Estimating the time of infection for African swine fever in pig farms in Korea.

African swine fever (ASF) is a highly contagious and lethal disease with characteristics of hemorrhagic fever. ASF outbreaks in pig farms significantly damage the entire pork industry. Understanding the transmission dynamics of ASF is crucial to effectively respond. Notably, it is important to know when the infection started on the outbreak farm. This study aimed at establishing a procedure for estimating the time of infection on pig farms affected by the ASF outbreak in Korea. The protocol for sampling to detect ASF virus infection, the estimation of the time interval between infection and detection, and the estimation of the infection stage parameters for the simulation model were described. After infection, fattening sheds (9.8 days in median) had the longest detection time compared with pregnant (8.6 days) or farrowing sheds (8.0 days). The intervals were 8.8 days for farrow-to-finisher farms, 7.0 days for farrow-to-weaning farms, and 9.5 days for fattening farms. The findings of this study provide valuable insights into ASF outbreaks in pig farms thus, improving the disease control ability.

A suspected case of a multiple autoimmune syndrome in a poodle dog.

A 9-year-old castrated male poodle dog was presented with icterus, anorexia, and lethargy. The dog was diagnosed with hypothyroidism 1 month before and was treated with levothyroxine. Severe anaemia with spherocytes, positive saline agglutination test, and hyperbilirubinemia indicated immune-mediated haemolytic anaemia (IMHA). Therefore, immunosuppressive therapy with prednisolone, mycophenolate mofetil, and danazol was started. Although the IMHA was well controlled, during tapering of prednisolone, acute multiple joint swelling and oedema suspected immune-mediated polyarthritis occurred twice. First, clinical symptoms improved as the dosage of prednisolone increased. However, the dog showed severe adverse effects to the steroid. Second time, we added leflunomide as another immunosuppressant, and clinical signs of arthritis disappeared. About 3 weeks later, despite the immunosuppressive therapy, skin lesions resembling an autoimmune dermatologic disorder spread throughout the body. Addition of cyclosporine resolved the skin lesions. This is a case report of a dog showing several sporadic clinical signs related to multiple autoimmune syndromes and their management using different immunosuppressant drugs.

Combination of immunosuppressive drugs and allogeneic stem cell treatment in a dog with suspected nephrotic syndrome.

The case study aims to describe the nephrotic syndrome (NS) in a castrated 3-year-old male Cocker Spaniel dog. The patient arrived at the hospital with a loss of appetite and weakness. Skin oedema with ascites was observed along with hypoalbuminaemia, hypoproteinaemia, hyperlipidaemia, hypercholesterolaemia, and proteinuria (urine protein to creatinine ratio = 22.4). Based on these findings, the patient was diagnosed with NS, although a renal biopsy was not conducted. Prednisolone (1 mg/kg, p.o. q12 h) and mycophenolate mofetil (10 mg/kg, p.o. q12 h) were prescribed as the immunosuppressive drugs, and previously cryopreserved allogeneic adipose tissue-derived mesenchymal stem cells (2 × 107 cells/kg) were injected intravenously. After several weeks of treatment, the patient recovered from NS. This is the first case report on immunosuppressive drugs and allogeneic mesenchymal stem cells being used to treat a dog with NS.

Macrophage PPAR-γ suppresses long-term lung fibrotic sequelae following acute influenza infection.

Influenza virus causes a heterogeneous respiratory infectious disease ranging from self-limiting symptoms to non-resolving pathology in the lungs. Worldwide, seasonal influenza infections claim ~500,000 lives annually. Recent reports describe pathologic pulmonary sequelae that result in remodeling the architecture of lung parenchyma following respiratory infections. These dysfunctional recovery processes that disproportionately impact the elderly have been understudied. Macrophages are involved in tissue remodeling and are critical for survival of severe influenza infection. Here, we found intrinsic deficiency of the nuclear receptor PPAR-γ in myeloid cells delayed the resolution of pulmonary inflammation following influenza infection. Mice with myeloid cell-specific PPAR-γ deficiency subsequently presented with increased influenza-induced deposition of pulmonary collagen compared to control mice. This dysfunctional lung remodeling was progressive and sustained for at least 3 months following infection of mice with myeloid PPAR-γ deficiency. These progressive changes were accompanied by a pro-fibrotic gene signature from lung macrophages and preceded by deficiencies in activation of genes involved with damage repair. Importantly similar aberrant gene expression patterns were also found in a secondary analysis of a study where macrophages were isolated from patients with fibrotic interstitial lung disease. Quite unexpectedly, mice with PPAR-γ deficient macrophages were more resistant to bleomycin-induced weight loss whereas extracellular matrix deposition was unaffected compared to controls. Therefore PPAR-γ expression in macrophages may be a pathogen-specific limiter of organ recovery rather than a ubiquitous effector pathway in response to generic damage.

A smartphone fluorescence imaging-based mobile biosensing system integrated with a passive fluidic control cartridge for minimal user intervention and high accuracy.

A key challenge for realizing mobile device-based on-the-spot environmental biodetection is that a biosensor integrated with a fluid handling sensor cartridge must have acceptable accuracy comparable to that of conventional standard analytical methods. Furthermore, the user interface must be easy to operate, technologically plausible, and concise. Herein, we introduced an advanced smartphone imaging-based fluorescence microscope designed for Hg2+ monitoring by utilizing a biosensor cartridge that reduced user intervention via time-sequenced passive fluid handling. The cartridge also employed a metal-nanostructured plastic substrate for complementing the fluorescence signal output; this helped the realization of high-accuracy detection, in which a ratiometric dual-wavelength detection method was applied. Using 30 samples of Hg2+-spiked wastewater, we showed that our device, which has a detection limit of ∼1 pM, can perform analytical assays accurately. The detection results from our method were in good linearity and agreement with those of conventional standard methods. We conclude that the integration of a simple-to-use biosensor cartridge, fluorescence signal-enhancing substrate, dual-wavelength detection, and quantitative image data processing on a smartphone has great potential to make any population accessible to small-molecule detection, which has been performed in centralized laboratories for environmental monitoring.

A smartphone imaging-based label-free and dual-wavelength fluorescent biosensor with high sensitivity and accuracy.

The accuracy of a bioassay based on smartphone-integrated fluorescent biosensors has been limited due to the occurrence of false signals from non-specific reactions as well as a high background and low signal-to-noise ratios for complementary metal oxide semiconductor image sensors. To overcome this problem, we demonstrate dual-wavelength fluorescent detection of biomolecules with high accuracy. Fluorescent intensity can be quantified using dual wavelengths simultaneously, where one decreases and the other increases, as the target analytes bind to the split capture and detection aptamer probes. To do this, we performed smartphone imaging-based fluorescence microscopy using a microarray platform on a substrate with metal-enhanced fluorescence (MEF) using Ag film and Al2O3 nano-spacer. The results showed that the sensitivity and specificity of the dual-wavelength fluorescent quantitative assay for the target biomolecule 17-ß-estradiol in water were significantly increased through the elimination of false signals. The detection limit was 1pg/mL and the area under the receiver operating characteristic curve of the proposed assay (0.922) was comparable to that of an enzyme-linked immunosorbent assay (0.956) from statistical accuracy tests using spiked wastewater samples. This novel method has great potential as an accurate point-of-care testing technology based on mobile platforms for clinical diagnostics and environmental monitoring.

The inhibitory activity of ginsenoside Rp4 in adenosine diphosphate-induced platelet aggregation.

BACKGROUND: Korean ginseng, Panax ginseng Meyer, has been used as a traditional oriental medicine to treat illness and promote health for several thousand years. Ginsenosides are the main constituents for the pharmacological effects of P. ginseng. Since several ginsenosides, including ginsenoside (G)-Rg3 and G-Rp1, have reported antiplatelet activity, here we investigate the ability of G-Rp4 to modulate adenosine diphosphate (ADP)-induced platelet aggregation. The ginsenoside Rp4, a similar chemical structure of G-Rp1, was prepared from G-Rg1 by chemical modification. METHODS: To examine the effects of G-Rp4 on platelet activation, we performed several experiments, including antiplatelet ability, the modulation of intracellular calcium concentration, and P-selectin expression. In addition, we examined the activation of integrin αIIbß3 and the phosphorylation of signaling molecules using fibrinogen binding assay and immunoblotting in rat washed platelets. RESULTS: G-Rp4 inhibited ADP-induced platelet aggregation in a dose-dependent manner. We found that G-Rp4 decreased calcium mobilization and P-selectin expression in ADP-activated platelets. Moreover, fibrinogen binding to integrin αIIbß3 by ADP was attenuated in G-Rp4-treated platelets. G-Rp4 significantly attenuated phosphorylation of extracellular signal-regulated protein kinases 1 and 2, p38, and c-Jun N-terminal kinase, as well as protein kinase B, phosphatidylinositol 3-kinase, and phospholipase C-γ phosphorylations. CONCLUSION: G-Rp4 significantly inhibited ADP-induced platelet aggregation and this is mediated via modulating the intracellular signaling molecules. These results indicate that G-Rp4 could be a potential candidate as a therapeutic agent against platelet-related cardiovascular diseases.

Seesawed fluorescence nano-aptasensor based on highly vertical ZnO nanorods and three-dimensional quantitative fluorescence imaging for enhanced detection accuracy of ATP.

Probe-mediated fluorescence biosensing methods based on spectrophotometry still have limitations such as detection inaccuracy caused by the occurrence of false signals and lack of simultaneous qualitative and quantitative read-outs with an ultra-low detection limit. Herein, we describe a novel seesawed fluorescence detection strategy based on dual-colour imaging-based quantitation in which the green fluorescence of the capture aptamer decreases and the red fluorescence of the detection aptamer increases simultaneously upon their respective interactions with the target biomolecule. This approach enhances detection accuracy through facilitating identification of probable false-positives in biological samples. Furthermore, combining the seesawed detection scheme with three-dimensional imaging of fluorescence signal enhanced by highly vertical ZnO nanorods increases signal-to-noise ratio, which addresses the limited performance of digital cameras and, in turn, enhances sensitivity and dynamic range. This simple, robust, scalable, imaging-based and label-free fluorescence method allows highly specific and sensitive quantification of biomolecules with excellent reliability.

Real-time label-free quantitative fluorescence microscopy-based detection of ATP using a tunable fluorescent nano-aptasensor platform.

Although real-time label-free fluorescent aptasensors based on nanomaterials are increasingly recognized as a useful strategy for the detection of target biomolecules with high fidelity, the lack of an imaging-based quantitative measurement platform limits their implementation with biological samples. Here we introduce an ensemble strategy for a real-time label-free fluorescent graphene (Gr) aptasensor platform. This platform employs aptamer length-dependent tunability, thus enabling the reagentless quantitative detection of biomolecules through computational processing coupled with real-time fluorescence imaging data. We demonstrate that this strategy effectively delivers dose-dependent quantitative readouts of adenosine triphosphate (ATP) concentration on chemical vapor deposited (CVD) Gr and reduced graphene oxide (rGO) surfaces, thereby providing cytotoxicity assessment. Compared with conventional fluorescence spectrometry methods, our highly efficient, universally applicable, and rational approach will facilitate broader implementation of imaging-based biosensing platforms for the quantitative evaluation of a range of target molecules.

Acetyl Eburicoic Acid from Laetiporus sulphureus var. miniatus Suppresses Inflammation in Murine Macrophage RAW 264.7 Cells.

The basidiomycete Laetiporus sulphureus var. miniatus belongs to the Aphyllophorales, Polyporaceae, and grows on the needleleaf tree. The fruiting bodies of Laetiporus species are known to produce N-methylated tyramine derivatives, polysaccharides, and various lanostane triterpenoids. As part of our ongoing effort to discover biologically active compounds from wood-rotting fungi, an anti-inflammatory triterpene, LSM-H7, has been isolated from the fruiting body of L. sulphureus var. miniatus and identified as acetyl eburicoic acid. LSM-H7 dose-dependently inhibited the NO production in RAW 264.7 cells without any cytotoxicity at the tested concentrations. Furthermore it suppressed the production of proinflammatory cytokines, mainly inducible nitric oxide synthase, cyclooxygenase-2, interleukin (IL)-1ß, IL-6 and tumor necrosis factor α, when compared with glyceraldehyde 3-phosphate dehydrogenase. These data suggest that LSM-H7 is a crucial component for the anti-inflammatory activity of L. sulphureus var. miniatus.

Cutting edge: progesterone directly upregulates vitamin d receptor gene expression for efficient regulation of T cells by calcitriol.

The two nuclear hormone receptor ligands progesterone and vitamin D (vit.D) play important roles in regulating T cells. The mechanism that connects these two hormones in regulating T cells has not been established. In this study, we report that progesterone is a novel inducer of vit.D receptor (VDR) in T cells and makes T cells highly sensitive to calcitriol. At the molecular level, the induction by progesterone is mediated by two progesterone receptor-binding elements in the intron region after the first noncoding exon of the human VDR gene. Increased expression of VDR by progesterone allows highly sensitive regulation of T cells by vit.D even when vit.D levels are suboptimal. This novel regulatory pathway allows enhanced induction of regulatory T cells but suppression of Th1 and Th17 cells by the two nuclear hormones. The results have significant ramifications in effective regulation of T cells to prevent adverse immune responses during pregnancy.

Association between nocturnal/supine hypertension and restless legs syndrome in patients with Parkinson's disease.

BACKGROUND AND OBJECTIVE: Autonomic disturbances and sleep problems are common non-motor symptoms in patients with Parkinson's disease (PD). Orthostatic hypotension, supine hypertension (SH), and nocturnal hypertension (NH) are inter-related in patients with PD. These abnormalities might be associated with restless legs syndrome (RLS), which occurs predominantly at rest or during sleep. Few reports have suggested an association between circadian blood pressure disturbances and RLS in the general population. We evaluated the relationship between neurocardiovascular blood pressure alterations and RLS in patients with early PD. METHODS: A total of 225 patients, newly diagnosed with PD, were included in the study. RLS was diagnosed by the International Restless Legs Syndrome Study Group's diagnostic criteria. Orthostatic vital signs and ambulatory 24-h blood pressure were monitored and recorded. RESULTS: Thirty-six (16.0%) participating patients had RLS. SH and NH were more frequent in the PD+RLS group than in the group without RLS. Supine blood pressure, orthostatic decline in blood pressure, nighttime blood pressure, and the standard deviation of systolic blood pressure were significantly higher in the PD+RLS group than in the group without RLS. CONCLUSION: RLS is related to nocturnal/supine hypertension and blood pressure fluctuations, suggesting a neuropathological association between autonomic and sleep dysfunctions in patients with PD. RLS may be a determinant of neurocirculatory abnormalities. Detecting and effectively treating RLS might slow the rate of pressure-related neurocardiovascular damage in dysautonomic patients with PD.

Photodynamic Therapy for Bowen's Disease of the Vulva Area.

Bowen's disease is a squamous cell carcinoma in situ and has the potential to progress to a squamous cell carcinoma. The authors treated two female patients (a 39-year-old and a 41-year-old) with Bowen's disease in the vulva area using topical photodynamic therapy (PDT), involving the use of 5-aminolaevulinic acid and a light-emitting diode device. The light was administered at an intensity of 80 mW/cm(2) for a dose of 120 J/cm(2) biweekly for 6 cycles. The 39-year-old patient showed excellent clinical improvement, but the other patient achieved only a partial response. Even though one patient underwent a total excision 1 year later due to recurrence, both patients were satisfied with the cosmetic outcomes of this therapy and the partial improvement over time. The common side effect of PDT was a stinging sensation. PDT provides a relatively effective and useful alternative treatment for Bowen's disease in the vulva area.

Causality assessment of cutaneous adverse drug reactions.

BACKGROUND: Cutaneous adverse drug reactions (ADRs) are the most common adverse reactions attributed to drugs. A systematic and effective approach to a patient with suspected drug eruption allows for prompt recognition, classification and treatment of cutaneous ADRs. A standardized and effective approach for objective causality assessment is necessary to make consistent and accurate identification of ADRs. OBJECTIVE: Although the Naranjo algorithm is the most widely used assessment tool, it contains many components which are not suitable for clinical assessment of ADRs in Korea. The purpose of this study is to compare correlations of the Naranjo algorithm and the Korean algorithm to evaluate usefulness of both algorithms in order to make a causal link between drugs and cutaneous ADRs. In addition, this study classifies the clinical types and causative agents of cutaneous ADRs. METHODS: The authors retrospectively reviewed the clinical types and laboratory findings of patients who were diagnosed with cutaneous ADRs in the dermatology clinic at Gil hospital. One hundred forty-one patients were enrolled in this evaluation. The causal relationship of ADRs was assessed by using the Naranjo algorithm and Korean algorithm (version 2.0). RESULTS: A cross-tabulation analysis was applied to the Naranjo algorithm and Korean algorithm (version 2.0). Simple correlation analysis and a Bland-Altman plot were used for statistical analysis. Correlation analysis confirmed that the two assessment algorithms were significantly correlated. Exanthematous eruptions (68.8%), Stevens- Johnson syndrome (10.6%), and urticaria (8.5%) were the most common types of cutaneoues ADRs. The most common causative agents were antibiotics/antimicrobials, antipyretics/non-steroidal anti-inflammatory drugs, and central nervous system depressants. CONCLUSION: The Naranjo algorithm and Korean algorithm (version 2.0) were significantly correlated with each other, and thus reliable assessment methods to determine cutaneous ADRs.

Photosensitivity reactions to vandetanib: redevelopment after sequential treatment with docetaxel.

Vandetanib (ZD6474, Zactima™) is a novel, orally available inhibitor of different intracellular signaling pathways involved in tumor growth, progression, and angiogenesis, including vascular endothelial growth factor receptor-2, epidermal growth factor receptor, and rearranged during transfection tyrosine kinase activity. The most frequently reported adverse events attributed to vandetanib include diarrhea, elevated aminotransferase, asymptomatic corrected QC interval prolongation, and hypertension. In a few randomized, double-blinded studies, cutaneous adverse events including these general symptoms have been reported, but there are only a few reports on the photosensitivity reaction to vandetanib domestically as conducted by dermatologists. In this report, we describe two cases of photosensitivity reactions induced by vandetanib. After improvement with steroid and antihistamine, the photosensitivity reaction was redeveloped by sequential treatment with docetaxel.

The neumann type of pemphigus vegetans treated with combination of dapsone and steroid.

Pemphigus vegetans is a rare variant of pemphigus vulgaris and is characterized by vegetating lesions in the inguinal folds and mouth and by the presence of autoantibodies against desmoglein 3. Two clinical subtypes of pemphigus vegetans exist, which are initially characterized by flaccid bullae and erosions (the Neumann subtype) or pustules (the Hallopeau subtype). Both subtypes subsequently develop into hyperpigmented vegetative plaques with pustules and hypertrophic granulation tissue at the periphery of the lesions. Oral administration of corticosteroids alone does not always induce disease remission in patients with pemphigus vegetans. We report here on a 63-year-old woman with pemphigs vegetans. She had a 2-year history of vegetating, papillomatous plaques on the inguinal folds and erosions of the oral mucosa. The enzyme-linked immunosorbent assay was positive for anti-desmoglein 3, but it was negative for anti-desmoglein 1. She was initially treated with systemic steroid, but no improvement was observed. The patient was then successfully treated with a combination of systemic steroid and dapsone with a good clinical response.

Chromoblastomycosis Caused by Phialophora richardsiae.

Chromoblastomycosis is a chronic fungal disease of the skin and subcutaneous tissues caused by a group of dematiaceous (black) fungi. The most common etiologic agents are Fonsecaea pedrosoi and Cladophialophora carrionii, both of which can be isolated from plant debris. The infection usually follows traumatic inoculation by a penetrating thorn or splinter wound. Several months after the injury, painless papules or nodules appear on the affected area; these papules then progress to scaly and verrucose plaques. We report a case of chromoblastomycosis caused by Phialophora richardsiae, which has been rarely associated with chromoblastomycosis. The case involved a 43-year-old male, who for the past 2 months had noted an erythematous, pustulous plaque that was somewhat dark brown in color on his right shin; the plaque also had intermittent purulent discharge and crust formation. On histopathological examination, chronic granulomatous inflammation and sclerotic cells were seen. The tissue fungus culture grew out the typical black fungi of P. richardsiae, which was confirmed by polymerase chain reaction. The patient has been treated with a combination of terbinafine and itraconazole for 3 months with a good clinical response.