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Background Systemic sclerosis (SSc) causes progressive fibrosis of multiple organs with the low efficacy of immunosuppressive therapies. Our previous study indicated the SSc pathological pathways are closely correlated with Ca2+ signals, and blockage of the intracellular Ca2+ elevation facilitates inhibition of SSc pathogenesis. Objective Transforming growth factor β (TGF-β)-modulated SMAD signaling is crucial in regulating SSc pathogenesis. Whether Ca2+ signals are involved in TGF-β1/SMAD signaling-induced fibrotic process has been further investigated. Methods We utilized TGF-β1-induced myofibroblasts as a model to detect how Ca2+ signals affected SSc pathogenesis, and investigated the combination of treatment with store-operated Ca2+ entry (SOCE) associated inhibitors, 2-aminoethyl diphenylborinate (2-APB) and SKF96365 to restrain the increased Ca2+ signaling in myofibroblasts. In addition, the SSc bleomycin mouse model was used to detect the effect of 2-APB on SSc pathogenesis in vivo. Results Our findings revealed increased levels of TGF-β1 production in SSc was associated with intracellular Ca2+ activity, and inhibition of intracellular Ca2+ regulation by 2-APB resulted in the dedifferentiation of TGF-β1-induced myofibroblasts. This was due to the fact that 2-APB restrained the expression fibrotic markers, α-SMA, fibronectin and vimentin through inhibiting TGF-β1/SMAD3 signaling. Thus, subcutaneous injection of 2-APB improved bleomycin-induced skin and pulmonary fibrosis. Conclusion 2-APB is a potential candidate for treating fibrosis, by disrupting intracellular Ca2+ regulation in SSc to induce the dedifferentiation of myofibroblasts and meliorates fibrosis pathogenesis via inhibiting TGF-β1/SMAD3 signaling.

Human breast milk lipids have major beneficial effects: they promote infant early brain development, growth and health. To identify the relationship between human breast milk lipids and infant neurodevelopment, multivariate analyses that combined lipidomics and psychological Bayley-III scales evaluation were utilized. We identified that 9,12-octadecadiynoic acid has a significantly positive correlation with infant adaptive behavioral development, which is a crucial neurodevelopment to manage risk from environmental stress. To further clarify the biological function of 9,12-octadecadiynoic acid in regulating neurodevelopment, Caenorhabditis elegans (C. elegans) was used as a model to investigate the effect of 9,12-octadecadiynoic acid on neurobehavioral development. Supplementation with 9,12-octadecadiynoic acid from the L1 to L4 stage in larvae affected locomotive behaviors and foraging ability that were not socially interactive, implying that 9,12-octadecadiynoic acid is involved in regulating the serotonergic neuronal ability. We found that supplementary 0.1 μM 9,12-octadecadiynoic acid accelerated the locomotive ability and foraging ability via increasing the expression of serotonin transporter mod-1. Antioxidant defense genes, sod-1, sod-3 and cyp-35A2 are involved in 9,12-octadecadiynoic acid-induced motor neuronal activity. Nevertheless, supplementary 9,12-octadecadiynoic acid at concentrations above 1 μM significantly attenuated locomotive behaviors, foraging ability, serotonin synthesis, serotonin-related gene expressions and stress-related gene expression, resulting in the decreased longevity of worms in the experiment. In conclusion, our study demonstrates the biological function of 9,12-octadecadiynoic acid in governing adaptive behavioral development.

Systemic sclerosis (SSc) causes progressive fibrosis of multiple organs with the low efficacy of immunosuppressive therapies. Our previous study indicated the SSc pathological pathways are closely correlated with CaTransforming growth factor β (TGF-β)-modulated SMAD signaling is crucial in regulating SSc pathogenesis. Whether CaWe utilized TGF-β1-induced myofibroblasts as a model to detect how CaOur findings revealed increased levels of TGF-β1 production in SSc was associated with intracellular Ca2-APB is a potential candidate for treating fibrosis, by disrupting intracellular Ca

Porcine circovirus type 3 (PCV3) is a newly emerging porcine circovirus that infects pig populations worldwide. In this study, we investigated the prevalence of PCV3 in Taiwan and analyzed the phylogenetic relationships between the Taiwanese PCV3 strains and those from other countries. A total of 463 clinical specimens from sick pigs were collected in 2016-2019 and analyzed for PCV3 by PCR. The positivity rate for PCV3 was 10.6% in 2016, increasing markedly to 34.78% in 2019. A phylogenetic analysis based on full-length genomic sequences of PCV3 divided the PCV3 strains into three clades, with the Taiwanese strains in clade 1.

Specific ethnic genetic backgrounds are associated with the risk of Stevens-Johnson syndrome / toxic epidermal necrolysis (SJS/TEN) especially in Asians. However, there have been no large cohort, multiple-country epidemiological studies of medication risk related to SJS/TEN in Asian populations. Thus, we analyzed the registration databases from multiple Asian countries who were treated during 1998-2017. A total 1,028 SJS/TEN cases were identified with the algorithm of drug causality for epidermal necrolysis. Furthermore, those medications labeled by the US Food and Drug Administration (FDA) as carrying a risk of SJS/TEN were also compared with the common causes of SJS/TEN in Asian countries. Oxcarbazepine, sulfasalazine, COX-II inhibitors, and strontium ranelate were identified as new potential causes. In addition to sulfa drugs and beta-lactam antibiotics, quinolones were also a common cause. Only one acetaminophen-induced SJS was identified, while several medications (e.g., oseltamivir, terbinafine, isotretinoin, and sorafenib) labeled as carrying a risk of SJS/TEN by the FDA were not found to have caused any of the cases in the Asian countries investigated in this study.

Based on the oxidative stress theory, aging derives from the accumulation of oxidized proteins induced by reactive oxygen species (ROS) in the cytoplasm. Hydrogen peroxide (H2O2) elicits ROS that induces skin aging through oxidation of proteins, forming disulfide bridges with cysteine or methionine sulfhydryl groups. Decreased Ca2+ signaling is observed in aged cells, probably secondary to the formation of disulfide bonds among Ca2+ signaling-related proteins. Skin aging processes are modeled by treating keratinocytes with H2O2. In the present study, H2O2 dose-dependently impaired the adenosine triphosphate (ATP)-induced Ca2+ response, which was partially protected via co-treatment with β-mercaptoethanol, resulting in reduced disulfide bond formation in inositol 1, 4, 5-trisphosphate receptors (IP3Rs). Molecular hydrogen (H2) was found to be more effectively protected H2O2-induced IP3R1 dysfunction by reducing disulfide bonds, rather than quenching ROS. In conclusion, skin aging processes may involve ROS-induced protein dysfunction due to disulfide bond formation, and H2 can protect oxidation of this process.

Aging, cancer, and longevity have been linked to intracellular Ca2+ signaling and nociceptive transient receptor potential (TRP) channels. We found that TRP canonical 7 (TRPC7) is a nociceptive mechanoreceptor and that TRPC7 channels specifically mediate the initiation of ultraviolet B (UVB)‐induced skin aging and tumor development due to p53 gene family mutations. Within 30 min after UVB irradiation, TRPC7 mediated UVB‐induced Ca2+ influx and the subsequent production of reactive oxygen species in skin cells. Notably, this function was unique to TRPC7 and was not observed for other TRP channels. In TRPC7 knockout mice, we did not observe the significant UVB‐associated pathology seen in wild‐type mice, including epidermal thickening, abnormal keratinocyte differentiation, and DNA damage response activation. TRPC7 knockout mice also had significantly fewer UVB‐induced cancerous tumors than did wild‐type mice, and UVB‐induced p53 gene family mutations were prevented in TRPC7 knockout mice. These results indicate that TRPC7 activity is pivotal in the initiation of UVB‐induced skin aging and tumorigenesis and that the reduction in TRPC7 activity suppresses the UVB‐induced aging process and tumor development. Our findings support that TRPC7 is a potential tumor initiator gene and that it causes cell aging and genomic instability, followed by a change in the activity of proto‐oncogenes and tumor suppressor genes to promote tumorigenesis., Schematic representation of the role of TRPC7 as a potential tumor initiator gene in ultraviolet B (UVB)‐induced aging and cancer progression. UVB‐activated TRPC7 initiates skin aging via intracellular Ca2+ elevation, resulting in oxidative stress, DNA damage response activation, abnormal differentiation, and senescence inflammation response activation; this pathology is repaired with the activation of p53 to maintain tissue homeostasis. When homeostasis is no longer maintained and the aging process is activated, carcinogens promote cancer progression. TRPC7 initiates tumorigenesis by causing genomic instability, thereby changing the activity of proto‐oncogenes and tumor suppressor genes.

Pseudorabies virus (PRV) is an alphaherpesvirus that causes pseudorabies (PR), an economically important viral disease of pigs. Marker vaccines were widely used in PR prevention and eradication programs. The purpose of this study was to construct a novel recombinant virus with deletions at defined regions in the glycoprotein E (gE) and thymine kinase (TK) genes by homologous recombination. This study also evaluated the safety and efficacy of the virus for a live attenuated marker vaccine. No significant difference was observed in virus replication between gE gene-deleted (gE(-)), gE/TK double gene-deleted (gE(-)TK(-)), and wild-type PRV by growth curve analysis. However, gE(-)TK(-) PRV was completely attenuated in mice. To evaluate the immunogenicity of gE(-)TK(-) PRV, four 12-week-old specific-pathogen-free pigs per group were immunized intramuscularly with viral titers of 1×10(4), 1×10(5), or 1×10(6) TCID50, followed by intranasal challenge infection with virulent PRV (1×10(8) TCID50) at 3 weeks post vaccination. The gE(-)TK(-) PRV-vaccinated pigs displayed no general adverse effects after immunization and had protective immune responses after PRV challenge. Thus, gE(-)TK(-) PRV was safe and efficacious and might be a potential candidate for a live attenuated marker vaccine against PRV.

Scrub typhus is caused by the obligate intracellular bacterium Orientia tsutsugamushi. Macrophages are host cells for its replication and clearance. Severe complications in patients are mainly caused by a cytokine storm resulting from overproduction of proinflammatory cytokines; nevertheless, the molecular mechanism for the occurrence remains obscure. Herein, we investigate the interactive regulation of cytokines and micro-RNA (miR) in human macrophages infected with low and high doses of O. tsutsugamushi. During low dose infection, macrophages produce high levels of IL-10 through extracellular signal-regulated kinase activation, which inhibits proinflammatory cytokine production and facilitates pathogen replication. Increasing levels of pathogen results in reduced levels of IL-10, and macrophages begin to generate high levels of proinflammatory cytokines through NF-κB activation. However, during a high dose infection, macrophages produce high levels of miR-155 to slow the proinflammatory response. The extracellular signal-regulated kinase/IL-10 axis suppresses the NF-κB/tumor necrosis factor alpha axis via activation of signal transducer and activator of transcription 3. Both IL-10 and miR-155 inhibit the NF-κB signaling pathway. Furthermore, IL-10 is a potent inhibitor of miR-155. Patients susceptible to a cytokine storm, peripheral blood mononuclear cells showed significantly lower IL-10 and miR-155 responses to O. tsutsugamushi challenge. Thus, IL-10 and miR-155 operate inhibitory mechanisms to achieve a proper defense mechanism and prevent a cytokine storm.

Syndecan-1 is a member of the transmembrane heparan sulfate proteoglycan family, whose membrane-bound and soluble forms are involved in wound healing, inflammation and vascular biology. Because these physiological events are implicated in the pathogenesis of systemic sclerosis (SSc), we investigated the clinical association of serum syndecan-1 levels in this disease. Serum syndecan-1 levels were significantly higher in SSc patients, both in diffuse cutaneous SSc (dcSSc) and limited cutaneous SSc (lcSSc), than in healthy individuals, while comparable between dcSSc and lcSSc groups. In late stage dcSSc patients (disease duration of >6 years), but not non-late stage dcSSc patients (≤6 years), serum syndecan-1 levels were significantly higher than in normal controls. More importantly, SSc patients with elevated serum syndecan-1 levels had higher prevalence of telangiectasia, elevated right ventricular systolic pressure and decreased diffuse capacity of the lung for carbon monoxide than those with normal levels. Therefore, soluble syndecan-1 may be related to the development of proliferative vasculopathy in SSc patients.

Aims The purpose of this study was to produce a recombinant pseudorabies virus (PRV) glycoprotein E (gE) protein with the correct antigenicity for use as a low-cost diagnostic antigen. Methods and Results The gene fragment encoding the amino-terminal immunodominant region of PRV gE (codons 31–270) (gEN31–270) was codon-optimized and expressed constitutively and secreted using a Pichia pastoris expression system. Yeast-expressed gEN31–270 (ygEN31–270) was harvested from the culture supernatant, and ygEN31–270 was shown to exhibit N-linked glycosylation. An indirect sandwich enzyme-linked immunosorbent assay (ELISA) was developed using ygEN31–270 as a coating antigen, and the results showed that the assay had high sensitivity and specificity, as well as almost perfect concordance with a commercial gE ELISA kit. Conclusions The immunodominant region (amino acids 31–270) of gE was expressed successfully in P. pastoris using a codon optimization strategy. ygEN31–270 was secreted and N-glycosylated. The ygEN31–270-based indirect sandwich ELISA showed high sensitivity and specificity to detect gE-specific antibodies in swine serum samples. Significance and impact of Study The ygEN31–270-based indirect sandwich ELISA may provide an alternative method for developing a diagnostic kit with easy manipulation and low cost. This article is protected by copyright. All rights reserved.

Background Bullous pemphigoid (BP) is a chronic blistering disease that manifests as multiple tense bullae on the limbs and body. Detecting biomarkers present in skin fluids may assist in the early diagnosis and treatment of BP. In this study, a modern mass spectrometric method was developed for screening biomarkers in blister fluids collected from patients. Methods Blister fluids collected from BP patients and physically injured patients were analyzed and compared using matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS). The blister fluids were mixed with MALDI matrix solution on the target plate; after drying, they were analyzed by MALDI-TOF MS. Results Alpha-defensins 1–3 were detected in the samples collected from all BP patients and absent in all patients with physical injuries. Therefore, alpha-defensins 1–3 are potential biomarkers for BP and can be used to differentiate between blisters caused by BP and those caused by physical injuries. Compared to traditional skin biopsy methods that use immunofluorescent stains, analyzing biomarkers in blister fluids using MALDI-TOF is a more rapid and less invasive method. Conclusions MALDI-TOF-MS is a non-invasive and efficient method that is able to rapidly distinguish between blisters caused by BP and those caused by physical injuries.

Porcine circovirus type 2 (PCV2) is the primary causative agent of an economically important swine disease, now known as porcine-associated disease (PCVAD). The only structural protein of viral capsid, Cap has become the major target for development of PCV2 subunit vaccines. The purpose of this study is to express Cap of PCV2 using a recombinant pseudorabies virus (PRV) that is gE gene deficient, which is a widely used PRV marker vaccine. The recombinant PRV, gE(-)/PCV2cap(+)PRV, was constructed using homologous recombination techniques, in order to replace the upstream of the gE gene with the PCV2 cap gene. The expression of Cap during virus replication was confirmed using immunofluorescence and Western blotting analysis. The expressed Cap protein self-assembled into virus-like particles (VLPs), which was demonstrated using electromicrography. The immunization of mice or guinea pigs with purified VLPs could induce significant, specific antibody responses to PCV2 Cap. These results demonstrate an alternative to PCV2 for the development of a VLP-based subunit vaccine.

Objective:To investigate the risk and genetic association of oxcarbazepine-induced cutaneous adverse reactions (OXC-cADRs), including Stevens-Johnson syndrome/toxic epidermal necrolysis (SJS/TEN), in Asian populations (Chinese and Thai).Methods:We prospectively enrolled patients with OXC-cADRs in Taiwan and Thailand from 2006 to 2014, and analyzed the clinical course, latent period, drug dosage, organ involvement, complications, and mortality. We also investigated the carrier rate ofHLA-B*15:02andHLA-A*31:01of patients with OXC-cADRs and compared to OXC-tolerant controls. The incidence of OXC-SJS/TEN was compared with carbamazepine (CBZ)–induced SJS/TEN according to the nationwide population dataset from the Taiwan National Health Insurance Research Database.Results:We enrolled 50 patients with OXC-cADRs, including 20 OXC-SJS/TEN and 6 drug reaction with eosinophilia and systemic symptoms, of Chinese patients from Taiwan and Thai patients from Thailand. OXC-cADRs presented with less clinical severity including limited skin detachment (all ≦5%) and no mortality. There was a significant association betweenHLA-B*15:02and OXC-SJS (p= 1.87 × 10−10; odds ratio 27.90; 95% confidence interval [CI] 7.84–99.23) in Chinese and this significant association was also observed in Thai patients. The positive and negative predictive values ofHLA-B*15:02for OXC-SJS/TEN were 0.73% and 99.97%, respectively.HLA-A*31:01was not associated with OXC-cADRs. The incidence and mortality of OXC-SJS/TEN was lower than CBZ-STS/TEN in new users (p= 0.003; relative risk 0.212; 95% CI 0.077–0.584).Conclusions:Our findings suggest thatHLA-B*15:02is significantly associated with OXC-SJS in Asian populations (Chinese and Thai). However, the severity and incidence of OXC-SJS/TEN are less than that of CBZ-SJS/TEN. The need for preemptiveHLA-B*15:02screening should be evaluated further.

The aim of this study was to investigate osteoporosis risk in atopic dermatitis (AD) patients. This study included patients in the Taiwan National Health Insurance Research dataset. The population-based study included all patients aged 20-49 years who had been diagnosed with AD during 1996-2010. In total, 35,229 age and gender-matched patients without AD in a 1:1 ratio were randomly selected as the non-AD group. Cox proportional-hazards regression and Kaplan-Meier analyses were used to measure the hazard ratios and the cumulative incidences of osteoporosis, respectively. During the follow-up period, 360(1.02%) AD patients and 127(0.36%) non-AD patients developed osteoporosis. The overall incidence of osteoporosis was4.72-fold greater in the AD patients compared to the non-AD patients (1.82 vs. 0.24 per 1,000 person-years, respectively) after adjusting for potential confounding factors. Osteoporosis risk factors included female gender, age, advanced Charlson Comorbidity Index, depression and use of corticosteroids. The dataset analysis showed that AD was significantly associated with subsequent risk of osteoporosis.

Orientia (O.) tsutsugamushi-induced scrub typhus is endemic across many regions of Asia and the Western Pacific, where an estimated 1 million cases occur each year; the majority of patients infected with O. tsutsugamushi end up with a cytokine storm from a severe inflammatory response. Previous reports have indicated that blocking tumor necrosis factor (TNF)-α reduced cell injury from a cytokine storm. Since TNF-α production is known to be associated with intracellular Ca2+ elevation, we examined the effect of store-operated Ca2+ entry (SOCE) inhibitors on TNF-α production in O. tsutsugamushi-infected macrophages. We found that 2-aminoethoxydiphenyl borate (2-APB), but not SKF96365, facilitates the suppression of Ca2+ mobilization via the interruption of Orai1 expression in O. tsutsugamushi-infected macrophages. Due to the decrease of Ca2+ elevation, the expression of TNF-α and its release from macrophages was repressed by 2-APB. In addition, a novel role of 2-APB was found in macrophages that causes the upregulation of heat shock protein 70 (HSP70) expression associated with ERK activation; upregulated TNF-α production in the case of knockdown HSP70 was inhibited with 2-APB treatment. Furthermore, elevated HSP70 formation unexpectedly did not help the cell survival of O. tsutsugamushi-infected macrophages. In conclusion, the parallelism between downregulated Ca2+ mobilization via SOCE and upregulated HSP70 after treatment with 2-APB against TNF-α production was found to efficiently attenuate an O. tsutsugamushi-induced severe inflammatory response.

Background: Oral cancer is a major public health burden in Taiwan. Studies have suggested that habits of betel quid chewing, tobacco smoking and alcohol drinking as well as their interaction effects are important risk factors for oral cancer and oral potentially malignant disorders (OPMD). A population-based oral cancer screening program has been implemented by Health Promotion Administration, Ministry of Health and Welfare, Taiwan. People with betel quid chewing and/or cigarette smoking habits are recommended for biennially screening. Previous studies found that majority of betel quid chewers were in labor intensive occupations. Given the fact that high risk groups of oral cancer are often reluctant to attend screening, targeted approach should be applied to improve the effectiveness of screening program. Aim: The aims of this study are to identify occupations with high prevalence of betel-quid chewing and also to investigate changing trends using the National Health Interview Survey (NHIS) database. Methods: The NHIS database of 2009 and 2013 were used to conduct this study. We identified current users of betel quid and/or cigarette from adults with aged 20-64 years old and working on jobs. Occupations were classified using the International Standard Classification of Occupations (ISCO-88). Statistical analyses included indirect standardized rate ratios and cluster analysis of chewing/smoking habits. Results: From the total survey participants (2009: 19,201, 2013: 17,249), we extracted 11,507 and 10,760 persons for the study. In the year of 2013, using the 2-digit code of ISCO-88, drivers and mobile-plant operators (code 83 in ISCO-88) had the highest prevalence of betel quid chewing habit (30.9%), followed by extraction and building trades workers (code 71 in ISCO-88, 29.6%). These two groups also had the 2 highest smoking prevalence (61.2% and 62.2%). The betel quid chewing prevalence decreased from 2009 to 2013 slightly (32.3%-30.9%). However, the prevalence of smoking increased (60.0%-62.2%). Among the betel quid chewers of the two focused occupations, the corresponding rates of receiving oral cancer screening were rather low (35.47% and 21.52%). The indirect standardized ratios using the total sample as the reference showed 6 occupations (2 digit of ISCO-88) with significantly higher ratios (larger than 2). Cluster analysis demonstrated that the group with high prevalence of betel quid chewing also had high prevalence of cigarette smoking habit. Conclusion: The betel quid chewing prevalence has been decreased in all occupations. While at the same time, prevalence of cigarette smoking did not largely decrease. We analyzed the NHIS databases and found that workers in some occupations tend to have higher prevalence in both betel quid chewing and smoking. The oral cancer screening and cancer awareness need be targeted on certain occupations.

Ultraviolet-B (UVB) is one of the most cytotoxic and mutagenic stresses that contribute to skin damage and aging through increasing intracellular Ca(2+) and reactive oxygen species (ROS). Derinat (sodium deoxyribonucleate) has been utilized as an immunomodulator for the treatment of ROS-associated diseases in clinics. However, the molecular mechanism by which Derinat protects skin cells from UVB-induced damage is poorly understood. Here, we show that Derinat significantly attenuated UVB-induced intracellular ROS production and decreased DNA damage in primary skin cells. Furthermore, Derinat reduced intracellular ROS, cyclooxygenase-2 (COX-2) expression and DNA damage in the skin of the BALB/c-nu mice exposed to UVB for seven days in vivo. Importantly, Derinat blocked the transient receptor potential canonical (TRPC) channels (TRPCs), as demonstrated by calcium imaging. Together, our results indicate that Derinat acts as a TRPCs blocker to reduce intracellular ROS production and DNA damage upon UVB irradiation. This mechanism provides a potential new application of Derinat for the protection against UVB-induced skin damage and aging.

A 7-year-old girl had a recalcitrant hyperkeratotic plaque on her left index finger, which had been present since the age of 3. She was referred to us for further management. During the past 4 years, she had received multiple sessions of liquid nitrogen treatment at monthly intervals. Because of intense pain and easy blistering after repeated cryotherapies, topical applications of Duofilm liquid (3% salicylic acid) and Aldara cream (5% imiquimod) were introduced; however, the verrucous plaque slightly reduced in size but regrew easily. Upon applying heavy pressure to the lesion site, hemorrhagic blisters frequently occurred. At her initial presentation, there were multiple brownish, verrucous papules aggregated on the coarse, indurated plaque on her left index finger (Figure 1A), indicating the typical appearance of verruca vulgaris. To identify the possible immunocompromised condition for this intractable lesion, common blood counts, immunoglobulin (Ig) G, IgM, and IgE were measured. These levels were all within normal limits, except for IgA, which was decreased (63.5 mg/dL; normal range 82e453 mg/dL). No signs of erosion or bone invasion were observed on an X-ray

Heparanase is an endo-β-D-glucuronidase cleaving heparan sulfate side-chains of heparin sulfate proteoglycans, which is involved in wound healing, inflammation, neovascularization and tumor progression through the degradation and remodeling of the extracellular matrix and the release of sequestered pro-angiogenic factors. Because heparanase-mediated biological processes seem to be involved in the development of systemic sclerosis (SSc), we investigated the clinical correlation of serum heparanase levels in patients with this disease. Serum heparanase levels were significantly higher in SSc patients than in healthy individuals, while comparable between diffuse cutaneous SSc and limited cutaneous SSc subgroups. On the other hand, SSc patients with digital ulcers had serum heparanase levels significantly lower than those without. These results suggest that serum heparanase levels may be elevated in SSc patients reflecting the contribution of heparanase-dependent biological processes to the development of SSc. SSc patients with high serum heparanase levels may be protected from the development of digital ulcers due to the increased release of sequestered pro-angiogenic factors such as vascular endothelial growth factor. Therefore, serum heparanase levels may serve as a protective marker against digital ulcers in SSc patients.

Arsenic-induced Bowen's disease (As-BD), a cutaneous carcinoma in situ, is thought to arise from gene mutation and uncontrolled proliferation. However, how mitochondria regulate the arsenic-induced cell proliferation remains unclear. The aim of this study was to clarify whether arsenic interfered with mitochondrial biogenesis and function, leading to aberrant cell proliferation in As-BD. Skin biopsy samples from patients with As-BD and controls were stained for cytochrome c oxidase (Complex IV), measured for mitochondrial DNA (mtDNA) copy number and the expression levels of mitochondrial biogenesis–related genes, including peroxisome proliferator–activated receptor gamma coactivator-1α (PGC-1α), nuclear respiratory factor 1 (NRF-1), and mitochondrial transcription factor A (mtTFA). The results showed that expression of cytochrome c oxidase, mtTFA, NRF-1, and PGC-1α was increased in As-BD compared with in healthy subjects. Treatment of primary keratinocytes with arsenic at concentrations lower than 1.0 μmol/L induced cell proliferation, along with enhanced mitochondrial biogenesis. Furthermore, we observed that the mitochondrial oxygen consumption rate and intracellular ATP level were increased in arsenic-treated keratinocytes. Blocking of mitochondrial function by oligomycin A (Complex V inhibitor) or knockdown of mtTFA by RNA interference abrogated arsenic-induced cell proliferation without affecting cyclin D1 expression. We concluded that mtTFA up-regulation, augmented mitochondrial biogenesis, and enhanced mitochondrial functions may contribute to arsenic-induced cell proliferation. Targeting mitochondrial biogenesis may help treat arsenical cancers at the stage of cell proliferation.

Background Atopic dermatitis (AD) is a chronic, relapsing dermatosis. Previous studies have focused mostly on pediatric patients, and investigations emphasizing adult AD have been limited. Objective We set out to determine the 1-year prevalence and evaluate the validity of the International Study of Asthma and Allergies in Childhood (ISAAC) and United Kingdom Working Party (UKWP) AD questionnaires of adult AD in Taiwan. Methods We conducted a cross-sectional study among nursing staff at a university hospital. The 1-year prevalence of AD was assessed by ISAAC and UKWP questionnaires. Subsequently, the dermatologists' diagnosis based on Hanifin and Rajka criteria was used as a reference for validation. Results The overall response rate was 92.9%, equivalent to 1131 complete questionnaires. Ninety adult patients with AD (8%) were identified by dermatologists' diagnosis whereas ISAAC identified 107 (9.5%); sensitivity and specificity were 36.7% and 92.9%, respectively. UKWP identified 42 (3.7%) patients with AD; sensitivity and specificity were 42.2% and 99.6%, respectively. Using the receiver operating characteristic curve analysis, the UKWP criteria performed significantly better than its ISAAC counterpart. Further analysis indicated that modification of these criteria resulted in significant improvement in their diagnostic efficacy. More specifically, modified ISAAC showed 90.0% and 55.2% sensitivity and specificity, respectively, whereas modified UKWP demonstrated 82.2% and 94.2% sensitivity and specificity, respectively. Limitation Most of the study subjects were female with a high educational background. Conclusion Currently available questionnaire instruments do not perform well in the identification of adult patients with AD. Modification of the original questionnaires may allow for future large-scale epidemiologic studies.

Pseudorabies virus (PRV) is an alphaherpesvirus, and its gene organization and regulation are similar to the well-characterized human herpes simplex virus (HSV). The PRV early protein UL54 consists of 363 amino acids with homology to the HSV ICP27 immediate-early protein. Previously, we have demonstrated the nuclear accumulation and poly(G) RNA-binding activity of UL54 protein. In the present study, we have identified further the functional regions within UL54 conferring for nuclear localization and RNA-binding activity. Several recombinant expression plasmids containing various coding regions of UL54 gene were constructed for producing a series of C-terminally truncated or internally deleted forms of UL54 mutants in Escherichia coli or porcine kidney (PK-15) cells. RNA-binding activity of E. coli-expressed UL54 mutants was characterized by the binding ability to poly(G) RNA homopolymer in dot blot hybridization assay and the results have shown that the N-terminal 83 residues were responsible for RNA-binding, and the region of residues 35-82 containing an RGG box was necessary for its function. Furthermore, the region responsible for nuclear localization was investigated by transient expression of various deletion mutants in PK-15 cells followed by detection of their subcellular distribution. The results showed that C-terminal deletion beyond the amino acid residue 83 or internal deletion containing the RGG box sequence could restrict UL54 mutants in the cytoplasm. The ability of the N-terminal 83 residues to target the green fluorescence protein to the nucleus confirmed further its role as a functional nuclear localization signal (NLS). The utmost N-terminal 83 residues portion of UL54 contains two important functional domains, NLS and RNA-binding, and thus it would play an indispensable role in UL54 regulatory function.

Burns are usually caused by fire; however, contact injuries after immersion in gasoline may also cause burn-like injuries to the skin.A 49-year-old man was trapped under an overturned tractor in a traffic accident for around 20 minutes. The oil tank was broken and his clothes were saturated as he lay in a prone position, partially immersed in a pool of leaded gasoline. There were no flames ignited at the accident. He was brought to the emergency department after more than half-an-hour.Initially there [...]

Pseudorabies virus (PRV) is an alphaherpesvirus, and its gene organization and regulation are similar to the well-characterized human simplex virus (HSV). Sequence analysis of the complete coding region of PRV UL54 gene revealed that the UL54 gene consisted of 1092 nucleotides encoding a protein of 363 amino acids and the gene showed homology to HSV immediate-early protein ICP27. Detection of the UL54 transcript in infected cells by reverse transcription-polymerase chain reaction (RT-PCR) demonstrated that the UL54 gene belonged to the early kinetic class based on sensitivity to cycloheximide and insensitivity to phosphonoacetic acid (PAA). To study the structure and function of UL54 protein, this gene was subcloned on Escherichia coli expression vector pET28b for overexpression, and the expressed product was applied to generate specific antibody against UL54 protein. The specificity of the mouse immuneserum was confirmed by its ability to react with a 40kDa viral protein present in the PRV infected cells in Western immunblotting assay, detected as early as 4h after infection. In addition, immunoperoxidasing staining of PRV infected cells undertaken with this antibody demonstrated mainly nuclear staining pattern. Furthermore, the RNA binding potential of UL54 protein was demonstrated by its binding activity to poly(G) RNA homopolymer in Northwestern blotting assay.

A multiplex PCR (mPCR) assay was developed and subsequently evaluated for its effectiveness as a means to simultaneously detect multiple viral infections of swine. Specific primers for each of four common DNA viruses, namely, pseudorabies virus (PRV), porcine circovirus type I (PCV1), porcine circovirus type II (PCV2), and porcine parvovirus (PPV), were used for testing procedure. The assay was shown to be highly sensitive in that as little as 10(-4) ng of each of the respective amplicons (approximately equal to 10,000 molecules) was detected when a composite of all four viruses (including both field and gene-deleted permutations of PRV) was tested as a single sample. It was also effective for detecting one or more of these same viruses in various combinations in specimens including lymph nodes, lungs, spleens, and tonsils collected from clinically ill pigs, and in specimens in spleen collected from aborted fetuses. The relative efficiency (compared to performing separate assays for each virus) and apparent sensitivity of mPCR suggest its potential application for routine molecular diagnostic purposes.