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Background: Increasing trends of HCV infection have been reported among HIV-positive men who have sex with men (MSM) in Europe, Australia and North America. The trends of recently acquired HCV infection among HIV-positive MSM are less clear in Asia–pacific region. Methods: All HIV-positive patients seeking care at a university hospital in Taiwan tested for anti-HCV IgG at least once annually to estimate the incidence of HCV seroconversion during 2011–2018. HCV genotyping and sequencing were performed and multivariate logistic regression analysis was conducted to identify the factors associated with HCV seroconversion among MSM. Findings: During the study period, 3495 HCV-seronegative patients (86·4% MSM) were included and 294 (8·4%) with recent HCV infection were identified, in whom 281 (95·6%) were MSM, during a total of 16,361·86 person-years of follow-up (PYFU), giving an overall incidence rate of 17·97 per 1000 PYFU, which increased from 14·28 per 1000 PYFU in 2011 to 25·38 per 1000 PYFU in 2018 (p

Cell culture remains as the golden standard for primary isolation of viruses in clinical specimens. In the current practice, researchers have to recognize the cytopathic effects (CPE) induced by virus infection and subsequently use virus-specific monoclonal antibody to confirm the presence of virus. Considering the broad applications of neural network in various fields, we aimed to utilize convolutional neural networks (CNN) to shorten the timing required for CPE identification and to improve the assay sensitivity. Based on the characteristics of influenza-induced CPE, a CNN model with larger sizes of filters and max-pooling kernels was constructed in the absence of transfer learning. A total of 601 images from mock-infected and influenza-infected MDCK cells were used to train the model. The performance of the model was tested by using extra 400 images and the percentage of correct recognition was 99.75%. To further examine the limit of our model in evaluating the changes of CPE overtime, additional 1190 images from a new experiment were used and the recognition rates at 16 hour (hr), 28 hr, and 40 hr post virus infection were 71.80%, 98.25%, and 87.46%, respectively. The specificity of our model, examined by images of MDCK cells infected by six other non-influenza viruses, was 100%. Hence, a simple CNN model was established to enhance the identification of influenza virus in clinical practice., Author summary Observation of cytopathic effects (CPE) induced by virus infection is a practical method to determine the prsence of viruses in the clinical specimens. However, CPE observation is labor-intensive and time-consuming because it requires medical examiner to inspect cell morphology changes for a period of time. Here, Convolutional Neural Networks (CNN) was applied to improve the disadvantage of CPE observation by using influenza virus as an example. To reduce the requirement for large image input of every clinical test, small amount of data was used to train our CNNs model without transfer learning and the trained model was examined with testing image data taken at 25hr post virus infection. The recognition of testing data shows that the model can identify CPE at 25hr and the high specificity of the model can differentiate the CPE induced by influenza viruses from those by other non-influenza viruses. The limit of our model was further examined by more experimental data of influenza-induced and mock-infected images, and the result shows our model can detect the slight changes at the initial stage of CPE development. Hence, our deep CNN model can significantly shorten the timing required to identify virus-induced cytopathic effects.

Hepatitis A virus (HAV) and hepatitis E virus (HEV) share the similar fecal-oral transmission route. During an outbreak of sexually transmitted acute hepatitis A among men who have sex with men (MSM) worldwide between 2015 and 2017, we investigated the possibility of sexual transmission and related morbidity of HEV infection among human immunodeficiency virus (HIV)-positive patients. From March 1, 2015 to August 31, 2017, anti-HEV immunoglobulin G was retrospectively determined among 3,293 HIV-positive patients, who were mainly MSM (87.6%) with a median CD4 count of 575 cells/μL. Prevalence and incidence of HEV infection were 3.7% (123 of 3,293) and 4.35 per 1,000 person-years of follow-up (PYFU), respectively, which were significantly lower compared with those of HAV infection (31.1% [996 of 3,204] and 12.61 per 1,000 PYFU, respectively). The number of patients with HEV infection did not increase with the hepatitis A epidemic. The factor associated with prevalent HEV infection was older age (per 1-year increase, adjusted odds ratio, 1.07; 95% confidence interval, 1.05-1.09), but neither sexual orientation nor acquisition of sexually transmitted infections was related to prevalent or incident HEV infection. Among 23 patients with incident HEV infection, 22 patients had viremia caused by HEV genotype 4. No patients had prolonged HEV viremia or clinical symptoms, and only a mild elevation of serum aminotransferase, ranging from 34 to 77 IU/L, was noted. Although 4 patients had hepatitis for a prolonged duration of between 8 and 17 months, no abdominal imaging revealed liver fibrosis or cirrhosis. Conclusion: HEV endemicity remained low among HIV-positive patients in Taiwan during the outbreak of acute hepatitis A. Our data suggest that sexual transmission of HEV with significant morbidity of HEV infection, if any, is rare in this population.

BACKGROUND Acute hepatitis C virus (HCV) infections have been increasingly reported among human immunodeficiency virus (HIV)-positive men who have sex with men (MSM) in the Asia-Pacific region. It remains unknown whether international network of HCV transmission has occurred in this region. METHODS HIV-positive patients with acute HCV infection, defined as HCV seroconversion within a year or documented acute hepatitis with seroconversion, diagnosed in Hong Kong, Taipei and Tokyo during 2010-2016 were included in this molecular epidemiology study. The NS5B region of the HCV genome (365 bp) was amplified using nested polymerase chain reaction and sequenced. RESULTS Of 234 HIV-positive patients with acute HCV infection, all were male with 94% being MSM. At the diagnosis of acute HCV infection, 73.5% had concurrent sexually transmitted diseases and 88.0% were receiving combination antiretroviral therapy. The most prevalent HCV genotype was 3a, 2a and 1b in Hong Kong, Taipei and Tokyo respectively. Nine independent clusters belonging to five genotypes (1b, 2a, 2c, 3a and 6a) were identified, each of which occurred in one city without overlapping except for one 3a sequence from Taipei that was closely related genetically to the Hong Kong cluster. CONCLUSIONS No international network of HCV transmission was identified among HIV-positive patients in the three Asia-Pacific cities. The transmission dynamics of sexually acquired HCV differed by city, but the risk of intercity clustering should not be ignored.

Two novel highly oxygenated nortriterpenoids, schisarisanlactones A (1) and B (2), have been isolated from the fruits of Schisandra arisanensis, an endemic plant of Taiwan. Compounds 1 and 2 possess an unprecedented 5/5/7/5/5-fused pentacyclic ring system. The structures of both compounds were determined on the basis of spectroscopic analyses, especially 2D NMR and MS. A plausible biogenetic pathway of 1 was proposed. Schisarisanlactone A (1) showed significant anti-HIV activity.

Human immunodeficiency virus type 1 (HIV-1) circulating recombinant form (CRF) 07_BC has caused serious HIV-1 epidemics among injecting drug users (IDUs) in East Asia. Little is known about the characteristics of the virus and its impact on disease progression among the infected individuals. In this study, we compared immunological progression between 423 IDUs infected with CRF07_BC and 194 men who have sex with men (MSM) with primary subtype B infection, and a representative full-length CRF07_BC molecular clone, pCRF07_BC, was constructed to characterize the virus. We found that IDUs infected with CRF07_BC had significantly slower immunological progression in the Cox proportional hazards model (hazard ratio: 0.30; 95% confidence interval: 0.13-0.69; P=0.004). The constructed recombinant CRF07_BC viruses had a reduced processing of the Gag/Gag-Pol polyproteins, a decreased incorporation of Vpr in the virus particle, tethering of virus particles on the plasma membrane and decreased virus growth kinetics. These phenotypes are related to the unique 7-amino acid deletion in the p6 of CRF07_BC, since complementation of the 7-amino acid in pCRF07_BC could improve the defective phenotypes. In summary, compared with MSM infected with HIV-1 subtype B, IDUs infected with CRF07_BC had slower immunological progression, which is likely correlated with interference of virus particle maturation by the 7-amino acid deletion in p6.

Antiretroviral therapy containing an integrase strand transfer inhibitor (INSTI) plus two NRTIs has become the recommended treatment for antiretroviral-naive HIV-1-infected patients in the updated guidelines. We aimed to determine the prevalence of INSTI-related mutations in Taiwan. Genotypic resistance assays were performed on plasma from ARV-naïve patients (N = 948), ARV-experienced but INSTI-naive patients (N = 359), and raltegravir-experienced patients (N = 63) from 2006 to 2015. Major INSTI mutations were defined according to the IAS-USA list and other substitutions with a Stanford HIVdb score ≧ 10 to at least one INSTI were defined as minor mutations. Of 1307 HIV-1 samples from patients never exposed to INSTIs, the overall prevalence of major resistance mutations to INSTIs was 0.9% (n = 12), with an increase to 1.2% in 2013. Of these 12 sequences, 11 harboured Q148H/K/R, one Y143R, and none N155H. Of 30 sequences (47.6%) with INSTI-resistant mutations from raltegravir-experienced patients, 17 harboured Q148H/K/R, 8 N155H, and 6 Y143C/R. Other than these major mutations, the prevalence of minor mutations were 5.3% and 38.1%, respectively, in ARV-naive and raltegravir-experienced patients. The overall prevalence of INSTI mutations remains low in Taiwan. Surveillance of INSTI resistance is warranted due to circulation of polymorphisms contributing to INSTI resistance and expected increasing use of INSTIs.

The human immunodeficiency virus type 1 (HIV-1)-encoded RNA-binding protein Tat is known to play an essential role in viral gene expression. In the search for novel compounds to inhibit Tat transactivity, one coumarin derivative, BPRHIV001, was identified, with a 50% effective concentration (EC 50 ) against HIV-1 at 1.3 nM. BPRHIV001 is likely to exert its effects at the stage after initiation of RNAPII elongation since Tat protein expression and the assembly of the Tat/P-TEFb complex remained unchanged. Next, a reduction of the p300 protein level, known to modulate Tat function through acetylation, was observed upon BPRHIV001 treatment, while the p300 mRNA level was unaffected. A concordant reduction of phosphorylated Akt, which was shown to be closely related to p300 stability, was observed in the presence of BPRHIV001 and was accompanied by a decrease of phosphorylated PDPK1, a well-known Akt activator. Furthermore, the docking analysis revealed that the reduced PDPK1 phosphorylation likely resulted from the allosteric effect of interaction between BPRHIV001 and PDPK1. With strong synergistic effects with current reverse transcriptase inhibitors, BPRHIV001 has the potential to become a promising lead compound for the development of a novel therapeutic agent against HIV-1 infection.

Objectives: To describe the incidence of hypersensitivity to abacavir and frequency of human leucocyte antigen (HLA)-B*5701 in HIV-infected Taiwanese persons. Methods: Medical records of 337 HIV-infected Taiwanese in whom abacavir-containing combination antiretroviral therapy (CART) was prescribed from 1 May 2001 to 31 December 2006 were reviewed, and HLA typing of the patients was performed in 320 patients (232 receiving abacavir and 88 not receiving abacavir) with available blood samples. HLA class I and II polymorphisms were determined by PCR with specific primers. HLA-B*5701 was further confirmed by sequence-based typing. Results: Of the 337 patients, median CD4 count was 166.5cells/mm 3 (range, 1.0-1914.0) and 83 patients (24.6%) had AIDS-defining opportunistic infections. Thirty-eight patients (11.3%) discontinued abacavir within 6 weeks of starting abacavir-containing CART. Among them, 10 patients had successful abacavir re-challenge and another 11 patients had other specific reasons for abacavir discontinuation. Therefore, 14 patients (4.2%) were classified as cases in whom abacavir hypersensitivity could not be excluded, and 3 patients (0.9%) met the criteria of abacavir hypersensitivity. Of the 320 patients undergoing HLA typing, HLA-A02 was the most common allele and only one individual (0.3%) expressed HLA-B*5701. Along with some differences in allele distributions, there was a significant difference in the genetic frequency of HLA-B57 in our patients compared with those of previous studies in other Chinese populations. Conclusions: Abacavir hypersensitivity was less frequently encountered in HIV-infected Taiwanese initiating abacavir-containing CART than in Caucasians, which might be explained by the low frequency of the HLA-B*5701 allele.

Background. Superinfection with hepatitis D virus (HDV) may increase the risk for hepatitis flares and chronic hepatic complications in patients with chronic hepatitis B virus (HBV) infection. This retrospective observational study aimed to examine the incidence of and factors associated with recent HDV superinfection among individuals coinfected with human immunodeficiency virus (HIV) and HBV. Method. Anti-HDV immunoglobulin G (IgG) was sequentially determined in 375 HIV/HBV-coinfected patients to estimate the HDV incidence between 1992 and 2012. Plasma HDV and HBV loads and HBV surface antigen (HBsAg) levels were determined for the HDV seroconverters. A nested case-control study was conducted to identify the associated factors with HDV seroconversion. Phylogenetic analysis was performed using HDV sequences amplified from HDV seroconverters and HDV-seropositive patients at baseline. Results. During 1762.4 person-years of follow-up [PYFU], 16 patients seroconverted for HDV, with an overall incidence rate of 9.07 per 1000 PYFU, which increased from 0 in 1992–2001, to 3.91 in 2002–2006, to 13.26 per 1000 PYFU in 2007–2012 (P < .05). Recent HDV infection was associated with elevated aminotransferase and bilirubin levels and elevated rapid plasma reagin titers. Of the 12 patients with HDV viremia, 2 were infected with genotype 2 and 10 with genotype 4. HBsAg levels remained elevated despite a significant decline of plasma HBV DNA load with combination antiretroviral therapy that contained lamivudine and/or tenofovir. Conclusions. Our findings show that the incidence of recent HDV infection in HIV/HBV-coinfected patients increased significantly from 1992–2001 to 2007–2011, and was associated with hepatitis flares and syphilis.

As the host of the 2008 Summer Olympic Games, Beijing implemented a series of stringent, short-term air quality control measures to reduce the emissions of anthropogenic air pollutants. Large reductions in the daily average concentrations of primary pollutants, e.g., non-methane hydrocarbons (NMHCs) and nitrogen oxides (NOx) of approximately 50% were observed at the air quality observatory of Peking University. Nevertheless, high levels of ozone were present during the control period. Although anthropogenic precursors were greatly reduced, the meteorological conditions in summer, including high temperature and light flux, are conducive to the production of large amounts of biogenic isoprene, which is extremely reactive. The diurnal pattern of isoprene showed daily maximum mixing ratios of 0.83 ppbv at noon and a minimum at night, reflecting its primarily biogenic properties. Using the ratio of isoprene to vehicle exhaust tracers, approximately 92% of the daytime isoprene was estimated from biogenic sources, and only 8% was attributed to vehicular emissions. In terms of OH reactivity and the ozone formation potential (OFP), biogenic isoprene with its midday surge can contribute approximately 20% of the total OFPs and 40–50% of the total OH reactivities of the 65 measured NMHCs during the midday hours. The discrepancy between decreased precursor levels and the observed high ozone was most likely caused by a combination of many factors. The changes in the partition among the components of oxidation products (O3, NO2 and NOz) and the contribution of air pollutants from regional sources outside Beijing should be two primary reasons. Furthermore, the influences of biogenic isoprene as well as the non-linearity of O3-VOC-NOx chemistry are other major concerns that can reduce the effectiveness of the control measures for decreasing ozone formation. Although anthropogenic precursors were greatly reduced during the Olympic Games, sufficient biogenic isoprene and moderate NOx were still present in the conditions of high radiation flux and temperature during midday and early afternoon, which can still contribute a significant fraction of midday and early afternoon O3.

We describe the clinical course and virological characteristics of the first H7N9 influenza case in a Taiwanese patient; this patient had detectable viruses in the airway for 2 weeks, during which time an oseltamivir resistance-associated R292K mutation rapidly emerged. Anti-H7N9 antibody was detected 21 days after onset of symptoms, when H7N9 viral load declined significantly.

The novel triterpenoid is tested for in vitro inhibitory activity against the HIV virus and exhibits significant effects.

An outbreak of respiratory adenovirus infection in children was observed in northern Taiwan between November 2004 and February 2005. Using polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) to determine the serotype(s) of 172 adenovirus isolates in the outbreak period, we found that adenovirus type 3 (Ad3) was the predominant type (87.2%), followed by Ad2 (6.4%), Ad1 (4.1%), Ad7 (1.2%), Ad4 (0.6%), and Ad5 (0.6%). The genotype of Ad3 was analyzed for 15 isolates from the outbreak period by RFLP of the full-length genome. All these isolates belonged to genotype Ad3a2. Compared with the Ad3-infected patients in the baseline period, a significantly higher proportion of Ad3-infected patients in the outbreak period had severe infections (58.0% vs. 40.2%, P = 0.01), which included bronchopneumonia (28.7%), exudative tonsillitis (24.1%), and tonsillitis (16.1%). Moreover, patients with severe infections were significantly younger than those without (4.10 vs. 8.15 years, P < 0.001). In summary, our study demonstrated that Ad3 was the predominant serotype responsible for the respiratory adenovirus outbreak in northern Taiwan during 2004-2005 and was associated with severe infections in the outbreak period.

Herpes simplex virus type 1 (HSV-1) can establish latent infection in the nervous system and usually leads to life-threatening diseases in immunocompromised individuals upon reactivation. Treatment with conventional nucleoside analogue such as acyclovir is effective in most cases, but drug-resistance may arise due to prolonged treatment in immunocompromised individuals. In this study, we identified an in-use medication, digitoxin, which actively inhibited HSV-1 replication with a 50% effective concentration (EC(50)) of 0.05 microM. The 50% cytotoxicity concentration (CC(50)) of digitoxin is 10.66 microM and the derived selective index is 213. Several structural analogues of digitoxin such as digoxin, ouabain octahydrate and G-strophanthin also showed anti-HSV activity. The inhibitory effects of digitoxin are likely to be introduced at the early stage of HSV-1 replication and the virus release stage. The observation that digitoxin can inhibit acyclovir-resistant viruses further implicates that digitoxin represents a novel drug class with distinct antiviral mechanisms from traditional drugs.

Lung cancer is one of the most common causes of cancer deaths in the world, and the emergence of resistant lung cancer cells can lead to treatment failure. The development of new drugs to treat lung cancer, especially those resistant lung cancer cells, is urgent. Previously we identified that coumarin could inhibit replication of human lung cancer A549 cells. The efficacy of 284 coumarin derivatives on human lung cancer A549 cells was examined, and BPR2P001S0 was the most potent compound. BPR2P001S0 exhibited growth inhibition and apoptotic activity on A549 cells with an IC50 of 10 nM. A similar level of inhibitory effects was also observed in cisplatin-resistant A549 cells (IC50 = 12.5 nM). The ability of colony formation and migration capability of A549 cells decreased significantly in the presence of BPR2P001S0. In cell cycle analysis, BPR2P001S0 induced cell cycle arrest at the G0/G1 phase through down-regulation of cyclin D1 and Cdk4 protein expression. The expression of cyclinD1/Cdk4 regulatory protein, Akt, was also decreased. Subsequently, we found that the expression level of Ser241 phosphorated PDPK1 protein, an Akt activator in Akt pathway, was also reduced. By molecular docking analysis, we found that the reduced PDPK1 phosphorylation is likely resulted from the allosteric effect of interactions between BPRHIV001 and PDPK1. In conclusion, our results suggest that the anti-tumor activity of BPR2P001S0 is likely resulting from its ability to interfere with PDPK1 autophosphorylation at Ser241, which leads to reduced Akt protein and subsequent cyclin D1 degradation. Citation Format: Pi-Han Lin, Tzu-Ming Jao, Yu-Lin Hung, Hsing-Pang Hsieh, Ya-Chien Yang, Sui-Yuan Chang. BPR2P001S0, a Coumarin derivative, induced cell cycle arrest in A549 through inhibiting PDPK1 activity. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 1754. doi:10.1158/1538-7445.AM2015-1754

Background The fourth-generation human immunodeficiency virus (HIV) combination assay, which can simultaneously detect the presence of anti-HIV antibody and HIV antigen, has been shown to shorten the window period in HIV diagnosis compared with the third-generation HIV antibody immunoassay. This study was aimed to determine the performance of HIV combination assays in Taiwan, where the HIV-1 seroprevalence is 0.007% and HIV-2 infection has never been reported. Methods Performance of three fourth-generation HIV Ag/Ab combination assays (Dia.Pro, Wantai, and Bio-Rad) and one third-generation HIV Ab immunoassay (AxSYM HIV 1/2 gO) was assessed. Results A total of 152 specimens, including 86 confirmed HIV-seropositive and 66 HIV-seronegative samples, were used in the study. The sensitivity of four assays varied from 98.8% to 100%, and specificity varied from 98.5% to 100%. Performance of the 75 equivocal samples, the HIV status of which was confirmed later, in terms of negative prediction varied from 81.8% to 87.5%. The Bio-Rad and Dia.Pro assays exhibited higher sensitivity for the detection of p24 antigen among the three fourth-generation HIV combination assays. Conclusion The three fourth-generation HIV Ag/Ab combination assays exhibited better sensitivity, specificity, and negative prediction than the third-generation HIV Ab immunoassay.