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Abstract: Sequences of the partial 293-bp nuclear Z-chromosome-linked chromo-helicase binding protein (CHD-Z) and 729-bp mitochondrial cytochrome b (cyt b) genes were obtained from two species of the same family Pycnonotidae of Pycnonotus sinensis (P. sinensis, Chinese Bulbul) and Pycnonotus taivanus (P. taivanus, Taiwan Bulbul) distributed in Taiwan. A panel of 10 individuals (n =5 for each) with unknown relationship was used to characterize single nucleotide polymorphisms (SNPs) of these genes. We identified 2 and 10 SNPs in CHD-Z and cyt b loci, respectively. Frequency of SNPs was 10 per every 1465- and 729-bp on average. Pairwise nucleotide divergences of CHD-Z and cyt b genes among 10 specimens ranged from 0 to 0.0034 and 0 to 0.0055, respectively. Phylogenetic analysis suggested that the P. taivanus group assignment based on the CHD-Z and cyt b sequences is obviously very similar to P. sinensis. [Copyright &y& Elsevier]

Abstract: Background: High-throughput single nucleotide polymorphism (SNP) genotyping generates a huge amount of SNP data in genome-wide association studies. Simultaneous analyses for multiple SNP interactions associated with many diseases and cancers are essential; however, these analyses are still computationally challenging. Methods: In this study, we propose an odds ratio-based binary particle swarm optimization (OR-BPSO) method to evaluate the risk of breast cancer. Results: BPSO provides the combinational SNPs with their corresponding genotype, called SNP barcodes, with the maximal difference of occurrence between the control and breast cancer groups. A specific SNP barcode with an optimized fitness value was identified among seven SNP combinations within the space of one minute. The identified SNP barcodes with the best performance between control and breast cancer groups were found to be control-dominant, suggesting that these SNP barcodes may prove protective against breast cancer. After statistical analysis, these control-dominant SNP barcodes were processed for odds ratio analysis for quantitative measurement with regard to the risk of breast cancer. Conclusion: This study proposes an effective high-speed method to analyze the SNP–SNP interactions for breast cancer association study. [Copyright &y& Elsevier]

Abstract: Structural properties of GaN epilayers on wet-etched protruding and recess-patterned sapphire substrates (PSSs) have been investigated in detail using high-resolution double-crystal X-ray diffraction (DCXRD) and etch-pit density methods. The DCXRD results reveal various dislocation configurations on both types of PSSs. The etch pits of GaN on the recess PSS exhibit a regular distribution, i.e. less etch pits or threading dislocation density (TDD) onto the recess area than those onto the sapphire mesas. On the contrary, an irregular distribution is observed for the etch pits of GaN on the protruding PSS. A higher crystal quality of the GaN epilayer grown onto the recess PSS can be achieved as compared with that onto the protruding PSS. These data reflect that the GaN epilayer on the recess PSS could be a better template for the second epitaxial lateral overgrowth (ELOG) of GaN. As a result, the GaN epilayers after the ELOG process display the TDDs of around ∼106 cm−2. [Copyright &y& Elsevier]

Abstract: Genome-wide association analysis involved many single-nucleotide polymorphisms (SNPs) data is challenging mathematically and computationally. Hence, we propose the odds ratio-based discrete binary particle swarm optimization (OR-DBPSO) method that uses the OR as a new quantitative measure of disease risk among many SNP combinations with genotypes called “SNP barcode”. DBPSO are applied to generate SNP barcode, which computes the maximal difference of occurrence between the case and control groups, to predict disease susceptibility such as osteoporosis. Different SNP barcode patterns may occur several times in either low or high bone mineral density (BMD) groups. Our results showed that a DBPSO can effectively identify a specific SNP barcode with an optimized fitness value. SNP barcodes with a low fitness value will naturally be discarded from the population. A representative SNP barcode with a variable number of SNPs is processed to OR analysis to determine the maximum difference between the low and high BMD groups in statistics manner. Therefore, this paper introduces a powerful procedure to analyze disease-associated SNP–SNP interaction in genome-wide genes. [Copyright &y& Elsevier]

Abstract: Gene expression profiles, which represent the state of a cell at a molecular level, have great potential as a medical diagnosis tool. Compared to the number of genes involved, available training data sets generally have a fairly small sample size in cancer type classification. These training data limitations constitute a challenge to certain classification methodologies. A reliable selection method for genes relevant for sample classification is needed in order to speed up the processing rate, decrease the predictive error rate, and to avoid incomprehensibility due to the large number of genes investigated. Improved binary particle swarm optimization (IBPSO) is used in this study to implement feature selection, and the K-nearest neighbor (K-NN) method serves as an evaluator of the IBPSO for gene expression data classification problems. Experimental results show that this method effectively simplifies feature selection and reduces the total number of features needed. The classification accuracy obtained by the proposed method has the highest classification accuracy in nine of the 11 gene expression data test problems, and is comparative to the classification accuracy of the two other test problems, as compared to the best results previously published. [Copyright &y& Elsevier]

Background: Antioxidants such as N-acetylcysteine and probucol have been used to protect patients from contrast media-induced nephrotoxicity. The mechanisms underlying these protective effects are not well understood. We hypothesized that acetylcysteine and probucol alter the activity of endogenous antioxidant enzyme activity. Methods: Four weeks after induction of diabetes with streptozotocin, diabetic and nondiabetic rats were divided into three groups. Group 1 rats did not receive any antioxidant agents. Group 2 rats were treated with acetylcysteine and group 3 rats with probucol for 1 week before injection of the contrast medium diatrizoate (DTZ). Results: We found that diabetic rats had higher renal glutathione peroxidase (GPx) activity than normal rats. DTZ suppressed renal GPx activity significantly in both group 1 diabetic and normal rats. Interestingly, renal GPx activity in both diabetic and normal rats pretreated with acetylcysteine or probucol was not inhibited by DTZ. Renal superoxide dismutase (SOD) increased significantly in normal rats after DTZ injection, but not in diabetic rats. Finally, acetylcysteine or probucol did not significantly influence renal SOD. Conclusions: These findings suggest that the renal protective effects of acetylcysteine and probucol against contrast-induced oxidative stress and nephrotoxicity may be mediated by altering endogenous GPx activity. [Copyright &y& Elsevier]

Growth of InGaSb/AlSb high hole mobility quantum well field effect transistors (QW FETs) on Si substrates with a step-graded GaAsSb metamorphic buffer layer by molecular beam epitaxy is explored. With an optimized growth temperature for the InGaSb/AlSb QW, hole mobility of 770 cm 2 /V s and 3060 cm 2 /V s have been achieved at room temperature and 77 K, respectively. It is also found that the twins in the samples do not cause significant anisotropic behavior of the InGaSb QW FETs in term of gate direction. [ABSTRACT FROM AUTHOR]

Several marine drugs exert anticancer effects by inducing oxidative stress, which becomes overloaded and kills cancer cells when redox homeostasis is imbalanced. The downregulation of antioxidant signaling induces oxidative stress, while its upregulation attenuates oxidative stress. Marine drugs have miRNA-modulating effects against cancer cells. However, the potential antioxidant targets of such drugs have been rarely explored. This review aims to categorize the marine-drug-modulated miRNAs that downregulate their antioxidant targets, causing oxidative stress in anticancer treatments. We also categorize the downregulation of oxidative-stress-inducing miRNAs in antioxidant protection among non-cancer cells. We summarize the putative antioxidant targets of miRNA-modulating marine drugs by introducing a bioinformatics tool (miRDB). Finally, the marine drugs affecting antioxidant targets are surveyed. In this way, the connections between marine drugs and their modulating miRNA and antioxidant targets are innovatively categorized to provide a precise network for exploring their potential anticancer functions and protective effects on non-cancer cells. • The downregulation of antioxidant signaling induces oxidative stress. • The upregulation of antioxidant signaling attenuates oxidative stress. • This review categorizes marine-drugs, miRNAs, antioxidant targets, and ROS in anticancer treatments. • Antioxidant targets of miRNA-modulating marine drugs are retrieved bioinformatically. • Marine drugs, miRNA, and antioxidant targets are innovatively connected. [ABSTRACT FROM AUTHOR]

Abstract: Multilayers [FePt(1nm)/B(t nm)]10 (t=0.05–0.6) were alternately deposited on a glass substrate and subsequently annealed by the rapid thermal process (RTP) at 800°C for 3min. After RTP, FePt and B layers intermix to form the FePtB film with (001) texture. The ordering degree of FePt was slightly increased with doped B. The (Fe–Pt)100−x B x (x=0, 5, 10) films show perpendicular magnetization and the minor FeB phase was indexed in isotropic (Fe–Pt)100−x B x (x=30, 40, 60) films. By adding Ag into (Fe–Pt)95B5 film, the ordering degree was slightly increased in (Fe–Pt)95(B0.9Ag0.1)5 film. In (Fe–Pt)100−x B x (x=5, 10) and (Fe–Pt)95(B0.9Ag0.1)5 granular films, the intermixed B or Ag atoms were diffused among FePt grain boundaries to isolate and refine FePt grains uniformly with average grain sizes of 20, 15, and 6.7nm, respectively. [Copyright &y& Elsevier]

Abstract: Ethnopharmacological relavence: Physalis angulata is well-known in traditional Chinese medicine as a ingredient for various herbal formulation; also, it has been shown to exhibit anti-cancer and anti-inflammatory effects. In this study, the ability of P. angulata to inhibit tumor metastasis and angiogenesis was investigated. Materials and methods: Anti-proliferative activity of ethyl acetate extracts of P. angulata (PA extracts), was determined against human oral squamous carcinoma (HSC-3) and human umbilical vein endothelial cells (HUVECs) by trypan blue exclusion method. Wound-healing migration, trans-well invasion, Western blotting and chick chorioallantoic membrane assay were carried out to determine the anti-metastatic and anti-angiogenic effects of PA extracts in vitro and in vivo. Results: We demonstrated that at sub-cytotoxic concentrations of PA extracts (5–15μg/mL) markedly inhibited the migration and invasion of highly metastatic HSC-3 cells as shown by wound-healing repair assay and trans-well assay. Gelatin zymography assay showed that PA extracts suppressed the activity of matrix metalloproteinase (MMP)-9 and -2, and urokinase plasminogen activator (u-PA) in HSC-3 cells. In addition, Western blot analysis confirmed that PA extracts significantly decreased MMP-2 and u-PA protein expression in HSC-3 cells. Notably, PA extracts significantly augmented the expression of their endogenous inhibitors, including tissue inhibitors of MMP (TIMP-1 and -2), and plasminogen activator inhibitors (PAI-1 and -2). Further investigations revealed that non-cytotoxic concentration of PA extracts (5–15μg/mL) inhibited vascular endothelial growth factor (VEGF)-induced proliferation, and migration/invasion of HUVECs in vitro. PA extracts also suppressed the activity of MMP-9, but not MMP-2, in HUVECs. Further, we observed, PA extracts strongly suppressed neovessel formation in the chorioallantoic membrane of chick embryos in vivo. Conclusions: These results strongly support an anti-metastatic and anti-angiogenic activity of P. angulata that may contribute to the development of better chemopreventive agent for cancer and inflammation. [Copyright &y& Elsevier]

Abstract: Aims: To evaluate the possible herb–drug interaction of St. John''s wort (SJW) extracts with indinavir in rats and to set up a model for characterizing pre-systemic sites for the interactions between orally administered herbs and pharmaceuticals. Main Methods: The in vivo pharmacokinetic study and in situ single-pass intestinal perfusion model were employed in the research. Plasma indinavir concentration and cytochrome P450 3A activities were measured by high-pressure liquid chromatography and spectrophotometric assays, respectively. Key findings: Oral administration of either 150 or 300mg/day SJW for 15days significantly reduced indinavir plasma levels with certain pharmacokinetic parameter changes. The cytochrome P450 3A analysis suggested that this interaction was attributable to the induction of indinavir metabolism. Further perfusion study demonstrated that both small intestine and the liver contributed significantly to the reduction of indinavir bioavailability and was flow rate-dependent. Moreover, the small intestine was the major site for the pre-systemic metabolism of indinavir, whether with or without SJW pretreatment. Significance: Herb–drug pharmacokinetic interactions between SJW and indinavir can be clearly observed in the Wistar rat model. Particularly, the respective first-pass effect contributed by the small intestine and the liver could be differentiated and quantified. The application of the animal model to investigating herb–drug interactions or other relevant research purposes is envisioned. [Copyright &y& Elsevier]

Abstract: To test the applicability of isovolumic relaxation flow propagation velocity (IRFPV) as an indicator of relaxation, 38 patients undergoing cardiac catheterization for chest pain were included. The time constant of isovolumic left ventricular (LV) pressure decline (tau) had a significant correlation with the LV end-diastolic pressure, the peak negative dp/dt, the LV end-diastolic dimension, the fractional shortening, the late diastolic transmitral flow velocity, the early and late diastolic mitral annular velocities, the rapid filling flow propagation velocity, and the natural logarithms of heart rate, ejection fraction, LV end-systolic dimension and IRFPV (r = −0.773, p < 0.001). IRFPV was selected as the major independent determinant of tau, explaining 46% of its variance. In prediction of tau ≥42 ms, the sensitivity and specificity were 100% and 83% for IRFPV ≤104 cm/s, 77% and 67% for the rapid filling flow propagation velocity ≤50 cm/s, and 58% and 67% for the early diastolic mitral annular velocity ≤8 cm/s, respectively. In conclusion, our data suggest that IRFPV could be a better marker of LV relaxation than other parameters, as the rapid filling flow propagation velocity or the early diastolic mitral annular velocity. (E-mail: wcvoon@giga.net.tw) [Copyright &y& Elsevier]

• Provide a theoretical method is developed to investigate free vibration of rectangular thin plates fully immersed influids. • Rayleigh-Ritz method is applied to obtain the resonant frequency and mode shape of the rectangular plate immersed in fluid. • The effect on the resonant frequency of the plate for different depth of fluid and position of rectangular plate in the fluid. • We present the pressure and velocity of the fluid fields coupled with the rectangular plate. • We propose a modified meshing method for analyzing the fluid-structure interaction problem. In this paper, a theoretical method is developed to investigate the effect of a finite flow field on free vibration of a rectangular thin plate fully immersed in an incompressible and inviscid liquid. Rayleigh–Ritz method is applied to obtaining resonant frequencies and mode shapes of the wet plate, and different boundary conditions of a rectangular plate are discussed in this study, including fully clamped, free, and cantilever plates. The Galerkin method is utilized to deal with continuity conditions on the interface between plate and fluid and the beam method is applied to constructing mode shapes of a rectangular plate. Numerical results of the finite element method (FEM) are used to compare with the theoretical analysis on resonant frequencies and mode shapes and it shows the high accuracy of the proposed solution. In order to provide a convenient and accurate method to deal with the vibration characteristics of an immersed plate, the convergence of the Galerkin method is discussed and a modified computation method is presented. Finally, the effect of fluid's boundary on the resonating plate and the relation between the flow field and plate are investigated. Image, graphical abstract [ABSTRACT FROM AUTHOR]

Low ankle-brachial index (ABI) calculated using systolic blood pressure (SBP) is associated with poor prognosis. However, there is no study assessing ABI calculated using mean artery pressure (MAP) and diastolic blood pressure (DBP) in predicting mortality. Two cohort populations were enrolled. The first population comprised 379 patients (106 patients with angiography-proved peripheral artery disease (PAD) and 273 relative normal patients) to evaluate the best cutoff values of ABImbp and ABIdbp for prediction of PAD. The second population included 941 patients undergoing echocardiographic examinations to assess the ability of different ABIs in predicting mortality. ABIs were measured using an ABI-form device. The best cutoff values of ABImbp and ABIdbp for prediction of PAD were 0.92 and 0.88. In our second population, median follow-up to mortality was 93 months. There were 87 cardiovascular and 228 overall deaths. Multivariable analysis showed ABIsbp, ABImap, ABIdbp, ABIsbp <0.9, and ABImap <0.92 could predict overall and cardiovascular mortality (all p < 0.001). ABIdbp <0.88 could only predict CV mortality (p = 0.033). In a direct comparison of 6 multivariable models, the basic model consisting of significant variables in the univariable analysis plus ABImap <0.92 had the highest predictive value for overall and cardiovascular mortality (all p < 0.001). In a direct comparison of 6 multivariable models, the basic model + ABImap < 0.92 was the best model in predicting overall and cardiovascular mortality. Hence, calculation of ABI using MAP except SBP might provide extra benefit in survival prediction. Image 1 • Usefulness of ABI using mean arterial pressure (MAP) and diastolic blood pressure (DBP) to predict mortality is unknown. • We found the best cutoff values of ABImap and ABIdbp for prediction of peripheral artery disease were 0.92 and 0.88l. • Basic model + ABImap < 0.92 had the highest predictive value for overall and CV mortality than other models. • Calculation of ABImap except ABIsbp might provide an extra benefit in survival prediction. [ABSTRACT FROM AUTHOR]

Three new 9-methyladeninium diterpenoids, nakamusines A−C (1 − 3), along with four known analogous metabolites and a precursor of them were isolated from the Taiwanese sponge Agelas nakamurai. These new chemical structures were determined by extensive spectroscopic analysis, particularly HRESIMS and NMR data. The absolute configuration of nakamusine A (1) was deduced by the experimental circular dichroism (CD) and calculated electronic circular dichroism (ECD) spectra, is the 9-methyladeninium diterpenoid with a different configuration from the genus Agelas. Furthermore, 9-methyladenine (8) was isolated from Agelas sponge in this study, proving it to be an imperative precursor for the biosynthesis of 9-methyladeninium diterpenoids in Agelas sponge. The in vitro cytotoxicity, and antibacterial activity of these sponge natural products were also evaluated. Notably, (−)-agelasine C (4) and (−)-agelasine D (5) inhibited the growth of the microbial pathogen, Neisseria gonorrhoeae. Moreover, 4 and 5 showed cytotoxicity toward the human breast adenocarcinoma (MDA-MB-231) and oral squamous cell carcinoma (Ca9-22) with IC 50 values of 37.4 and 12.1 μM, respectively. [Display omitted] • Three new diterpenoids, nakamusines A−C, were isolated from sponge Agelas. • 9-Methyladenine is first discovery as precursor of 9-methyladeninium diterpenoids. • (−)-Agelasines C and D inhibit pathogens and show cytotoxicity to cancer cells. [ABSTRACT FROM AUTHOR]

A new cembranoid, flexibanone (1) , isolated from a Taiwanese soft coral Sinularia flexibilis , is the first cembranoid possessing an α-methylene-δ-lactone linking with a tetrahydronfuran ring by C-3/C-4 single bond. [Display omitted] A new cembranoid, flexibanone (1), and a known related diterpene sinulariolone (2) were isolated from a Taiwanese soft coral Sinularia flexibilis. Compound 1 is the first cembranoid possessing an α-methylene-δ-lactone linking with a tetrahydronfuran ring by C-3/C-4 single bond. The structures of 1 and 2 were established by using extensive spectroscopic analysis, including 2D NMR methods. Both compounds were suggested to be biosynthesized from the same cembranoid precursor. Further, the cytotoxicity against human oral cancer Ca9-22, breast adenocarcinoma MDA-MB-231 and non-small cell lung cancer A549 cell lines of 1 and 2 were also assayed. [ABSTRACT FROM AUTHOR]

Objective Sinuleptolide, a soft corals-derived bioactive norditerpenoid, is a marine natural product with a potent anti-inflammatory effect. We evaluate the potential anti-oral cancer effects of sinuleptolide and investigate the possible mechanisms involved. Designs Cell viability, cell cycle, apoptosis, reactive oxygen species (ROS), mitochondrial membrane potential (MMP), and DNA damage analyses were performed. Results In a cell viability assay, we found that sinuleptolide is dose-responsively antiproliferative against oral gingival cancer Ca9-22 cells but less harmful to normal human gingival fibroblast (HGF-1) cells ( P < 0.001). In cell cycle analysis, sinuleptolide induced subG1 accumulation at a higher dose and led to G2/M arrest of Ca9-22 cells ( P < 0.005). Apoptosis was significantly increased in sinuleptolide-treated Ca9-22 cells based on annexin V and poly(ADP-ribose) polymerase (PARP) expressions ( P < 0.05–0.0001). Based on flow cytometer analysis, sinuleptolide also induced the generation of ROS and decreased MMP in a dose-responsive manner ( P < 0.05–0.0001). DNA damage increased dose-responsively after sinuleptolide treatments ( P < 0.001) based on comet and γH2AX assays. Conclusion Sinuleptolide can induce an antiproliferation of oral cancer Ca9-22 cells involving apoptosis, oxidative stress and DNA damage, suggesting that sinuleptolide represents a potential chemotherapeutic drug for oral cancer treatment. [ABSTRACT FROM AUTHOR]

Epigenetic modifications play important roles in regulating carcinogenesis, and specific epigenetic modifications have emerged as potential tumor markers. Herein, we summarize several types of epigenetic modifications, explore the role played by epigenetic modifications in gene regulation, and describe the patterns of epigenetic modifications in cancers. Since epigenetic modifications have been reported to regulate the Warburg effect in cancers, the roles of epigenetic modifications in sugar metabolism are discussed. In addition, oxidative stress may play an important role in carcinogenesis, and the role of oxidative stress and epigenetic modification in carcinogenesis is addressed. We also discuss the role of epigenetic modifications as therapeutic targets. Finally, the synergistic effects of the combined treatment of epigenetic regulator and anticancer drugs for cancer therapy are described. [ABSTRACT FROM AUTHOR]

Abstract: Chronic dialysis association study involving individual single nucleotide polymorphisms (SNPs) in the mitochondrial displacement loop (D-loop) has previously been reported. However, possible SNP–SNP interactions for SNPs in the D-loop which could be associated with a reduced risk for chronic dialysis were not investigated. The purpose of this study was to propose an effective algorithm to identify protective SNP–SNP interactions in the D-loop from chronic dialysis patients. We introduce ISGA that uses an initialization strategy for genetic algorithms (GA) to improve the computational analysis for protective SNP–SNP interactions. ISGA generates genotype patterns with combined SNPs (SNP barcodes) for chronic dialysis. Using our previously reported 77 SNPs in the D-loop, the algorithm-generated protective SNP barcodes for chronic dialysis were evaluated. ISGA provides the SNP barcodes with the maximum frequency differences of occurrence between the cases and controls. The identified SNP barcodes with the lowest odds ratio (OR) values were regarded as the best preventive SNP barcodes against chronic dialysis. The best ISGA-generated SNP barcodes (two to nine SNPs) are more closely associated with the prevention of chronic dialysis when more SNPs are chosen (OR =0.64 to 0.32; 95% confidence interval=0.882 to 0.198). The cumulative effects of SNP–SNP interactions were more dominant in ISGA rather than in GA without the initialization strategy. We provide a fast identification of chronic dialysis-associated protective SNP barcodes and demonstrate that the SNP–SNP interactions may have a cumulative effect on prediction for chronic dialysis. [Copyright &y& Elsevier]

Summary: Objectives:: This analysis examined the association between oral hygiene and head and neck cancer (HNC) and whether this association differed by the consumption of alcohol, betel quid, or cigarette and by the genetic polymorphisms of inflammation-related genes. Materials and methods:: Interviews regarding dental care and oral health were conducted with 317 HNC cases and 296 controls. Genotyping was performed for 6 single nucleotide polymorphisms in IL6, IL10 and PTGS2. Results:: A positive association was observed between HNC and no regular dental visits (odds ratio (OR)=2.86, 95% confidence interval (CI): 1.47–5.57), brushing teeth <2times/day (OR=1.51, 95% CI: 1.02–2.23), frequent gum bleeding (OR=3.15, 95% CI: 1.36–7.28), and loss of >20 teeth (OR=2.31, 95% CI: 1.05–5.07). Analysis with dental care score (range: 0–4, 4=worst dental care), which combined regular dental visits, toothbrushing, and use of dental floss and mouthwash, showed a positive trend with HNC risk, particularly among alcohol drinkers and cigarette smokers. Multifactor dimensionality reduction analysis divided the study subjects into high- and low-risk group based on combinations of dental care score and IL6 rs1800796 genotypes. Compared to the low-risk group, the high-risk group had an OR of HNC=2.16 (95% CI: 1.44–3.25). Conclusions:: This study observed a positive association between poor oral hygiene and HNC, which appeared to differ by alcohol or cigarette consumption and the genotypes of IL6 rs1800796. Further investigations are needed to determine whether poor oral hygiene is a cause for HNC or a surrogatemarker of an unhealthy lifestyle that increases the risk of HNC. [Copyright &y& Elsevier]

A Pseudomonas exotoxin (PE-KDEL)-based chimeric subunit vaccine system was recently developed using a reverse vaccinology technique. In this study, the plasmids containing PE-PRRS chimeric subunits were constructed that composed of porcine reproductive and respiratory syndrome virus (PRRSV) antigen moieties, a ligand moiety and a Pseudomonas exotoxin A deleted domain III (PE (DIII)), and a carboxyl terminal moiety that includes a polypeptide with amino acid sequence KDEL (K3). The PE-PRRS combination vaccine can effectively induce not only PRRSV-specific INF-γ cellular immunity but also a slow-reacting and complement-requiring type serum neutralizing antibody in pigs. In a specific pathogen free (SPF) pig challenge model, body temperature (colonic temperature), occurrence of PRRSV viremia, nasal excretions, gross and histopathological appearances of pneumonia, and serum antibody activity (IFA and SN) titers significantly differed between the immunized group and the control group. The survey showed that a 0.3 mg/dose PE-PRRS vaccine formula conferred protection against PRRSV. A field trial of PE-PRRS vaccine was performed to study the immune response of pregnant sows after vaccination in a PRRSV persist farm. The RT-PCR analysis of viremia and serological titers showed that the PE-PRRS vaccine not only increased sow reproductive performance and evoked its immune response to PRRS viremia, it also activated maternal immune protections to prevent piglets from inflicting viremia. In conclusion, we developed a novel and effective PRRS cytotoxic T-cells (CTLs)-based vaccine containing Pseudomonas exotoxin (PE-KDEL) carrier in combination with PRRSV conserved epitopes against PRRS virus. [ABSTRACT FROM AUTHOR]

Summary: Objectives: We compared the outcomes and survival rates of patients with oral cavity squamous cell carcinoma receiving postoperative conventional radiotherapy (RT) or intensity-modulated radiotherapy (IMRT). Materials and methods: From January 2005 to September 2008, medical records of 131 consecutive patients with oral cancer receiving postoperative radiotherapy in the Department of Radiation Oncology of National Cheng Kung University Hospital were reviewed. Patients were divided into two groups according to the administration of postoperative conventional RT or IMRT. The loco-regional control, survival, and other prognostic factors were compared. Results: The 3-year Kaplan–Meier estimates of overall survival for patients receiving conventional RT and IMRT groups were 51.2% vs. 69.4% (p =0.079), respectively. The 3-year local–regional control (53.5% vs. 76.3%; p =0.020) and disease-free survival rates (47.8% vs. 70.0%; p =0.027) were significantly increased in the IMRT group. This retrospective study also identified that extracapsular spreading, margin positive/close (⩽2mm), more advanced T stage (T3–4 vs. T1–2), and conventional RT method were associated with worse prognosis. Conclusions: The addition of chemotherapy to adjuvant radiotherapy is recommended in patients with above risk factors. Our result underscores that IMRT should be considered to apply to OSCC patients referred for postoperative treatment. [ABSTRACT FROM AUTHOR]

Abstract: Grapeseed extract (GSE) is a rich source of natural phenolic compounds and possesses various pharmacological activities, including antioxidant, anticarcinogenic and anti-inflammatory properties. However, effects of GSE on immunoglobulin (Ig) E-mediated allergic responses still remain elusive. In the present study, the effects of GSE on activation and degranulation of RBL-2H3 mast cells were investigated. GSE pretreatments (20–100μg/ml) reduced IgE-antigen mediated release of β-hexosaminidase and histamine in RBL-2H3 cells. Additionally, GSE reversibly inhibited expression of FcεRI on RBL-2H3 cells. GSE treatments caused a significant elevation of intracellular cAMP levels, whereas the Ca2+ influx upon antigen stimulation was inhibited. Suppression on FcεRI expression together with decreased calcium uptake and increased cAMP level might be involved in attenuated degranulation of mast cells by GSE treatment. Our results suggest a possible pharmaceutical application of GSE in treating type I allergic diseases. [Copyright &y& Elsevier]

Abstract: Flavokawain B is a natural chalcone isolated from the rhizomes of Alpenia pricei Hayata. In the present study, we have investigated the antiproliferative and apoptotic effect of flavokawain B (5–20 μg/ml; 17.6–70.4 μM) against human squamous carcinoma (KB) cells. Exposure of KB cells with flavokawain B resulted in apoptosis, evidenced by loss of cell viability, profound morphological changes, genomic DNA fragmentation and sub-G1 phase accumulation. Apoptosis induced by flavokawain B results in activation of caspase-9, -3 and -8, cleavage of poly ADP ribose polymerase (PARP) and Bid in KB cells. Flavokawain B also down-regulate Bcl-2 with concomitant increase in Bax level, which resulted in release of cytochrome c. Taken together, the induction of apoptosis by flavokawain B involved in both death receptor and mitochondrial pathway. We also observed that flavokawain B caused the G2/M phase arrest that was mediated through reductions in the levels of cyclin A, cyclin B1, Cdc2 and Cdc25C and increases in p21/WAF1, Wee1 and p53 levels. Moreover, flavokawain B significantly inhibits matrix metalloproteinase-9 and urokinase plasminogen activator expression, whereas tissue inhibitor of matrix metalloproteinase-1 and plasminogen activator inhibitor-1 were increased, which are playing critical role in tumor metastasis. In addition, flavokawain B treatment significantly inhibited in vivo growth of human KB cell-derived tumor xenografts in nude mice, which is evidenced by augmentation of apoptotic DNA fragmentation, as detected by in situ terminal deoxynucleotidyl transferase-meditated dUTP nick end-labeling staining. The induction of cell cycle arrest and apoptosis by flavokawain B may provide a pivotal mechanism for its cancer chemopreventive action. [Copyright &y& Elsevier]

Purpose: Constitutively activated signal transducers and activators of transcription (STAT) factors, in particular STAT1, STAT3, and STAT5, have been detected in a wide variety of human primary tumors and have been demonstrated to directly contribute to oncogenesis. However, the expression pattern of these STATs in cervical carcinoma is still unknown, as is whether or not they have prognostic significance. This study investigated the expression patterns of STAT1, STAT3, and STAT5 in cervical cancer and their associations with clinical outcomes in patients treated with radical radiation therapy. Methods and Materials: A total of 165 consecutive patients with International Federation of Gynecology and Obstetrics (FIGO) Stages IB to IVA cervical cancer underwent radical radiation therapy, including external beam and/or high-dose-rate brachytherapy between 1989 and 2002. Immunohistochemical studies of their formalin-fixed, paraffin-embedded tissues were performed. Univariate and multivariate analyses were performed to identify and to evaluate the effects of these factors affecting patient survival. Results: Constitutive activations of STAT1, STAT3, and STAT5 were observed in 11%, 22%, and 61% of the participants, respectively. While STAT5 activation was associated with significantly better metastasis-free survival (p < 0.01) and overall survival (p = 0.04), STAT1 and STAT3 activation were not. Multivariate analyses showed that STAT5 activation, bulky tumor (≥4 cm), advanced stage (FIGO Stages III and IV), and brachytherapy (yes vs. no) were independent prognostic factors for cause-specific overall survival. None of the STATs was associated with local relapse. STAT5 activation (odds ratio = 0.29, 95% confidence interval = 0.13–0.63) and advanced stage (odds ratio = 2.54; 95% confidence interval = 1.03–6.26) were independent predictors of distant metastasis. Conclusions: This is the first report to provide the overall expression patterns and prognostic significance of specific STATs in cervical carcinoma. Our results indicate that constitutive STAT5 activation correlates with better metastasis-free survival and overall survival in cervical cancer patients who have received radiation therapy. [ABSTRACT FROM AUTHOR]

Abstract: The fermented culture broth of Antrodia camphorata (A. camphorata) has been shown to promote cell cycle arrest and apoptosis of human estrogen-nonresponsive MDA-MB-231 cells. Herein, we demonstrate that non-cytotoxic concentrations (20–80μg/mL) of A. camphorata markedly inhibited the invasion/migration of highly metastatic MDA-MB-231 cells as shown by an in vitro transwell and a wound-healing repair assay. The results of a gelatin zymography assay showed that A. camphorata suppressed the activity of matrix metalloproteinase (MMP)-9 and urokinase plasminogen activator (uPA). Western blot results demonstrated that treatment with A. camphorata decreased the expression of MMP-9, MMP-2, uPA, uPA receptor (uPAR) and vascular endothelial growth factor (VEGF); while the expression of the endogenous inhibitors of these proteins, i.e., tissue inhibitors of MMP (TIMP-1 and TIMP-2), and plasminogen activator inhibitor (PAI)-1, increased. Further investigation revealed that A. camphorata suppressed the phosphorylation of ERK1/2, p38, and JNK1/2. A. camphorata treatment also led to a dose-dependent inhibition on NF-κB binding and activation. This is the first report confirming the anti-metastatic activity of this potentially beneficial mushroom against human breast cancer. [Copyright &y& Elsevier]

Abstract: In this study, supercritical carbon dioxide extraction (SC-CO2) technology was developed to extract the active components (such as antioxidants) from a novel microalga, Chlorella vulgaris C-C, due to its superior advantages over conventional solvent or ultrasonic extraction methods. Using SC-CO2 and ultrasonic extractions, the polyphenol contents of C. vulgaris C-C were 13.40 and 0.46 (mggallic acid/g lyophilized extract), respectively. The flavonoid content obtained from SC-CO2 (3.18mgquercetin/g lyophilized extract) was also significantly higher than from ultrasonic extraction (0.86mgquercetin/g lyophilized extract). Investigation of C. vulgaris C-C extract from SC-CO2 indicates strong antioxidant activities in radical scavenging, ferric reducing power and metal chelating abilities. In cell proliferation assay, the extract of C. vulgaris C-C inhibits human lung cancer H1299, A549, and H1437 cells in a dose-dependent manner. In addition, the treatment with extracts of C. vulgaris C-C effectively reduced the migration of tumor cells, suggesting the potential of using the C. vulgaris C-C extract to inhibit lung cancer metastasis. [ABSTRACT FROM AUTHOR]

Abstract: Aims: The aims of the study are to investigate the additive effect of exogenous short-carbon chain phospholipids, C2-ceramide, on an anti-cancer drug paclitaxel (PTX)-induced senescence of human non-small cell lung cancer (NSCLC) cells deficient in functional p53 and p16, and to examine whether mitogen-activated protein kinase (MAPK) plays a role in ceramide-sensitized senescence of NSCLC cells. Main methods: To determine whether exogenous C2-ceramide renders lung cancer cells more sensitive to PTX treatment, techniques employing a flow cytometry-based cell cycle analysis and acidic β-galactosidase staining for senescent cells were used. Furthermore, to elucidate the role of MAPK proteins in modulating senescence, assays for protein levels of selective MAPKs and Bcl-2 family members, and detection of transcriptional levels senescence-associated genes were used in the study. Key findings: A sub-lethal dose of C2-ceramide sensitized the NSCLC H1299 cells to PTX treatment. The additive effects of C2-ceramide and PTX resulted in proliferative inhibition, G2-phase arrest of cell cycle, activation of p38 and eventually premature senescence. Importantly, neither p53, p21 waf1/cip1 nor p16 ink4 was shown to be involved in C2-ceramide-sensitized proliferative inhibition and senescence of H1299 cells by PTX in our study. Significance: Our study demonstrates that the short-carbon chain C2-ceramide can effectively sensitize PTX-induced senescence of H1299 cells via both p21 waf1/cip1 - and p16 ink4 -independent pathways. [Copyright &y& Elsevier]

Abstract: Mid-diastolic mitral annular motion may be driven by strain energy, an energy for myocardial recoil, stored during the previous systole. Hence, various patterns of mid-diastolic mitral annular motion may imply different left ventricular (LV) diastolic function. The purpose of this study is to compare LV diastolic properties among different types of mid-diastolic mitral annular motion. Two-hundred and three consecutive subjects underwent an echocardiographic examination at our outpatient clinic. Study subjects were classified into three groups according to mid-diastolic mitral annular motion patterns. Upward and downward La waves were defined, respectively, as a clear apically and atrially directed mid-diastolic annular motion on at least three consecutive beats with the average peak velocity ≥2 cm/s. Subjects with upward La wave, with downward but without upward La wave and without La wave were categorized as groups 1, 2 and 3, respectively. Early diastolic mitral annular velocity (Ea) was higher and the ratio of transmitral E wave velocity to Ea was lower in group 1 than in groups 2 and 3 (all p < 0.001). The diagnostic accuracy of upward La wave in prediction of normal diastolic function fell between 75% and 88%. In conclusion, patients with upward La wave had better LV diastolic function and lower LV filling pressure than patients without it. Upward La wave is useful in prediction of normal diastolic function. Therefore, analysis of mid-diastolic mitral annular motion may be complementary to other measures of LV diastolic function. (E-mail: wcvoon@giga.net.tw) [Copyright &y& Elsevier]

Abstract: Histone deacetylase inhibitors are promising chemotherapeutic agents and some are in clinical trials. Several molecular mechanisms have been invoked to describe their effects on cancer cells in vivo and in vitro. Autophagy has been observed in response to several anticancer reagents and has been demonstrated to be responsible for cell death. However, the exact mechanism of this phenomenon is still not clear. Here we demonstrated that suberoylanilide hydroxamic acid, a histone deacetylase inhibitor, induces nonapoptotic cell death with several specific features characteristic of autophagy in Hela S3 cells. Suberoylanilide hydroxamic acid inhibits the activity of the mammalian target of rapamycin, a negative regulator of macroautophagy which induces the formation of autophagosomes in a Beclin 1- and autophagy-related 7-dependent manner. This process is mediated by Akt and tuberous sclerosis 2 as is demonstrated by inhibition by continuous active Akt plasmid transfection and RNA interference of tuberous sclerosis 2. Our data provide the first evidence that suberoylanilide hydroxamic acid induces autophagy in Hela S3 cells through interference with the mammalian target of rapamycin signaling pathway. These findings suggest that suberoylanilide hydroxamic acid may induce autophagic cancer cell death via its specific pathway, and invite further investigation into the detailed mechanism of this pathway to explore this compound''s full potential as a chemotherapeutic agent. [Copyright &y& Elsevier]

Abstract: Background: Cardiac memory (CM) can be induced by both short and long period of pacing from the right ventricle. Although several mechanisms have been proposed in animal studies, mechanisms of CM in humans are not well studied. Methods: A total of 46 patients (20 females; mean age 46±13 years) with paroxysmal supraventricular tachycardia referred for catheter ablation were enrolled. After catheter ablation, CM was induced by 20 min of pacing from right ventricular apex (RVA). The CM was quantified as the difference of T wave area in each lead between baseline and after RVA pacing. After complete recovery from the induced CM, verapamil (1.5 mg/kg; 0.005 mg/kg/min), lidocaine (1 mg/kg; 2 mg/min), procainamide (10 mg/kg; 4 mg/min), and nitroglycerine (0.6 mg sublingually; 5 μg/min), were given in 14, 10, 12, and 10 patients respectively. The pacing procedure was repeated and the degrees of CM were compared before and after each drug administered. Results: The short-term CM was demonstrated by changes in T wave area after RVA pacing in all patients. The degrees of CM were suppressed in patients after verapamil and lidocaine. In contrast, procainamide and nitroglycerin had no significant effect on the degrees of CM expression. Conclusions: The expression of short-term CM can be suppressed by verapamil and lidocaine but not by procainamide and nitroglycerin. The results may suggest that short-term CM in humans can be modulated by calcium dependent process and the functional alternations of sodium and potassium channels. [Copyright &y& Elsevier]

Abstract: Background: Although the superior vena cava (SVC) may be involved in the triggering or maintenance of atrial fibrillation (AF), the electrophysiological properties of SVC in human are ill-defined. Methods: The baseline effective refractory periods (ERPs) of high right atrium (HRA), SVC and the conduction time (CT) between HRA and SVC were measured at pacing cycle lengths (PCL) of 600 and 400 ms respectively in 20 patients (12 females, age 46±13 years) with paroxysmal supraventricular tachycardia. Immediately after acute electrical remodeling (ER) induced by constant HRA or SVC pacing at PCL of 400 ms for 5 min, ERPs of HRA, SVC and the CT between HRA and SVC were determined. After verapamil was administered, the same protocols for determining ERPs of HRA, SVC and the CT between HRA and SVC were repeated. Results: The baseline ERP of SVC was significantly longer than that of HRA. The CT from SVC to HRA was significantly longer than that from HRA to SVC. After acute ER, both the ERPs of HRA and SVC were significantly shortened. However, no significant changes of the CT between HRA and SVC could be demonstrated. After verapamil infusion, significant shortening of the ERP of HRA and SVC still occurred following acute ER and the ERP of SVC was still longer than that of HRA. Conclusions: In human, ER can occur both in HRA and SVC after a short and moderately rapid heart rate pacing either from HRA or SVC. Verapamil cannot prevent such ER from occurring. [Copyright &y& Elsevier]

Abstract: Human herpesvirus 8 (HHV-8) is associated with the development of Kaposi''s sarcoma and several other human malignancies. Kaposin A protein of HHV-8 has been demonstrated as inducing tumorigenic transformation, being responsible for nuclear receptor coactivators in the transforming activity. In this study, a kaposin A-interacting septin 4 variant that contained the unique GDR at the N-terminus and AAALE at the C-terminus was identified using affinity selection of a phage display library. Co-immunoprecipitation and confocal imaging revealed in vitro binding specificity and in vivo co-localization of HHV-8 kaposin A protein to the septin 4 variant. The kaposin A-interacting septin 4 variant induced cell rounding up, activated caspase-3, and up-regulated transcriptional factor NF-κB. By contrast, kaposin A protein showed an antagonistic effect on the biological functions of the septin 4 variant. Therefore, the interaction of kaposin A protein and the septin 4 variant was suggested as playing a possible role in the development of HHV-8-associated malignancies. This study provides insights into the mechanism of the kaposin A protein pathology, in which the interactions of kaposin A protein with cellular proteins might allow alteration of fundamental cellular processes. [Copyright &y& Elsevier]

Diastolic mitral annular motion may terminate earlier in patients with higher left ventricular end-diastolic pressure (LVEDP). It was therefore hypothesized that the time interval measured from the end of the diastolic mitral annular velocity pattern to the onset of QRS (the AQ interval) would be a useful parameter in predicting LVEDP. The aim of this study was to evaluate the relation between the AQ interval and LVEDP. Forty-six patients with suspected coronary artery disease who underwent Doppler echocardiographic studies and cardiac catheterization were included. LVEDP was determined using a micromanometer-tipped catheter. On univariate analysis, the AQ interval had positive correlations with the PR interval (r = 0.405, p = 0.005), transmitral E-wave velocity (r = 0.502, p <0.001), isovolumic contraction time (r = 0.635, p <0.001), and LVEDP (r = 0.514, p <0.001) and a negative correlation with E-wave deceleration time (r = -0.430, p = 0.003). After stepwise multiple linear regression analysis, the PR interval, transmitral E-wave velocity, and LVEDP were the independent predictors of the AQ interval (beta = 0.234, p = 0.033; beta = 0.331, p = 0.004; and beta = 0.350, p = 0.003, respectively). In conclusion, the AQ interval is a novel, simple, and easily obtained index in the prediction of LVEDP. [ABSTRACT FROM AUTHOR]

Diastolic mitral annular motion may terminate earlier in patients with higher left ventricular end-diastolic pressure (LVEDP). It was therefore hypothesized that the time interval measured from the end of the diastolic mitral annular velocity pattern to the onset of QRS (the AQ interval) would be a useful parameter in predicting LVEDP. The aim of this study was to evaluate the relation between the AQ interval and LVEDP. Forty-six patients with suspected coronary artery disease who underwent Doppler echocardiographic studies and cardiac catheterization were included. LVEDP was determined using a micromanometer-tipped catheter. On univariate analysis, the AQ interval had positive correlations with the PR interval (r = 0.405, p = 0.005), transmitral E-wave velocity (r = 0.502, p <0.001), isovolumic contraction time (r = 0.635, p <0.001), and LVEDP (r = 0.514, p <0.001) and a negative correlation with E-wave deceleration time (r = −0.430, p = 0.003). After stepwise multiple linear regression analysis, the PR interval, transmitral E-wave velocity, and LVEDP were the independent predictors of the AQ interval (β = 0.234, p = 0.033; β = 0.331, p = 0.004; and β = 0.350, p = 0.003, respectively). In conclusion, the AQ interval is a novel, simple, and easily obtained index in the prediction of LVEDP. [Copyright &y& Elsevier]

Purpose: Radiotherapy is the most effective treatment for nasopharyngeal carcinoma (NPC). The aim of this study is to evaluate the efficacy and toxicity of fractionated stereotactic body radiation therapy (SBRT) boost for NPC. Methods and Materials: Sixty-four patients with newly diagnosed, nonmetastatic NPC were treated with conventional radiotherapy 64.8–68.4 Gy followed by fractionated SBRT boost 12–15 Gy between January 2002 and July 2004. Most patients (72%) presented with Stage III-IV disease. Fifty-two patients also received cisplatin-based concurrent (38) or neoadjuvant (14) chemotherapy. The major endpoints were local control, overall survival, and complications. Results: All patients finished the planned dose of radiotherapy. After a median follow-up of 31 months (range, 22–54), 15 patients developed tumor recurrences—3 in the nasopharynx, 4 in the neck, 5 in distant sites, 1 in both nasopharynx and neck, 2 in the neck and a distant site. The 3-year actuarial rate of local control was 93.1%, regional control 91.4%, freedom from distant metastasis 90.3%, and overall survival 84.9%, respectively. There were no Grade 4 acute or chronic radiation-related complications. Conclusions: Fractionated SBRT boost for NPC is technically feasible and provides good local control without any severe complications. [Copyright &y& Elsevier]

Abstract: During 2003–2004, approximately 13% of birth in Taiwan was given by foreign-born females. The aims of this study were to compare the seroprevalence of rubella antibodies between Taiwan-born and foreign-born pregnant women and evaluate the effect of rubella vaccination program in Taiwan. We reviewed the rubella antibody test results of 5007 women during routine pregnancy check-ups at Fooyin University Hospital during 1999–2002. In Taiwan-born women, rubella antibody was undetectable in 29.2%, 7.3%, and 8.3% of the cohorts born before 1971, between 1971 and 1976, and after 1976, respectively. In the cohorts born between 1971 and 1976 and after 1976, pregnant women born in China, Vietnam, Indonesia, and Philippines had significant higher chances of being susceptible. Our results suggested that the voluntary adult vaccination program was not as effective as the school or wipe-out programs. Both Taiwanese women born before 1971 and foreign-born women were more likely to be susceptible to rubella. The introduction of ‘catch-up’ immunization program and enforcement of the checking of immunization record and/or blood test before pregnancy for these women are necessary in preventing CRS. [Copyright &y& Elsevier]

Background: In addition to lowering lipid profile, statins have pleiotropic effects on improving vascular function. Changes of these pleiotropic effects and their relationship to lipid profile after withdrawal of statin treatment remained unclear. Methods: We investigated the changes of lipid profile and plasma concentrations of human soluble vascular cell adhesion molecule-1 (sVCAM-1) and human tissue type plasminogen activator (tPA) after withdrawal of statin treatment. Twenty-two patients (14 F, 8 M, aged 63±13 years) with hypercholesterolemia were treated with atorvastatin (ATOR; 10 mg/day) for 12 weeks. Blood samplings for serum lipid and markers were collected before, after and withdrawal of statin treatment. Results: The total cholesterol, LDL-cholesterol and sVCAM-1 (from 394.4±251.7 to 321.0±198.9 ng/ml, p<0.05) all decreased significantly and the tPA (from 9.47±3.57 to 11.62±3.99 ng/ml, p<0.05) increased significantly after atorvastatin treatment. During the following 3 days after withdrawal of atorvastatin, the total cholesterol and LDL-cholesterol did not show any significant change. However, the plasma sVCAM-1 significantly elevated on day 2 (from 321.0±198.9 to 371.2±220.4 ng/ml, p<0.05) and the plasma tPA significantly decreased on day 1 (from 11.62±3.99 to 10.52±3.55 ng/ml, p<0.05) and day 3 (from 11.62±3.99 to 10.27±3.69 ng/ml, p<0.05). Both the significant elevation of sVCAM-1 and decrease of tPA after withdrawal of atorvastatin treatment occurred within 3 days, while the serum cholesterol and LDL-chol levels did not have any significant change and were still within the therapeutic range. Conclusions: After 12 weeks of atorvastatin treatment, the beneficial pleiotropic effects can be demonstrated simultaneously with lowering the lipid profile. However, after withdrawal of atorvastatin, the beneficial pleiotropic effects are abrogated rapidly within days and are independent on elevation of serum cholesterol. [Copyright &y& Elsevier]

Abstract: The objective was to perform high-throughput gender identification of three Columbidae species (Columba livia, Columba pulchricollis, and Streptopelia tranquebarica). Although the chromo-helicase-DNA binding protein (CHD)-based Griffiths P2/P8 primer set resolved the amplicon products of these species in 3% agarose gel electrophoresis, it was unsuitable for molecular gender identification using the melting curve analysis (MCA) curve for high-throughput analysis. After sequencing the CHD-Z and CHD-W genes for these species, we redesigned a female-specific CHD-W primer (dove-W) and a female/male (or CHD-Z/CHD-W)-common primer (dove-ZW) to combine with the Griffiths P2 primer to generate two PCR amplicons with different lengths (P2/dove-W and P2/dove-ZW for 252- and 104-bp, respectively). Melting temperature (Tm) values for P2/dove-W and P2/dove-ZW amplicons were determined and resolved in MCA at approximately 79.0∼79.5 and 77.5 °C, respectively. Accordingly, females contained two Tm peaks, whereas males contained one. In conclusion, melting curve analysis (MCA) using our proposed primer sets was a robust gender identification method for the three Columbidae species tested. [ABSTRACT FROM AUTHOR]