Emerging evidence suggests that long noncoding RNA (lncRNA) plays pivotal roles in regulating various biological process in human cancers. Titin-antisense RNA1 (TTN-AS1) has been regarded as a tumor promoting lncRNA in numerous cancers. However, the clinical significance and biological function of TTN-AS1 in lung adenocarcinoma (LUAD) remain unclear. In the present study, we revealed that the expression of TTN-AS1 was upregulated in LUAD tissues and cell lines. High TTN-AS1 expression was associated with TNM stage and lymph node metastasis of LUAD patients. In addition, high expression of TTN-AS1 was correlated with poor postoperative prognosis of LUAD patients. Knockdown of TTN-AS1 significantly inhibited the growth, proliferation, migration, and invasion ability of LUAD cells in vitro. Then, by using bioinformation analysis and luciferase reporter experiment, we identified that TTN-AS1 could function as a competing endogenous RNA (ceRNA) by sponging miR-142-5p to regulate the expression of cyclin-dependent kinase 5 (CDK5) in LUAD. Since CDK5 is a key regulator in the process of epithelial mesenchymal transition (EMT), we detected the expression of EMT-related proteins, consequently, EMT was suppressed by knockdown of TTN-AS1 while this phenomenon was rescued by miR-142-5p inhibitor. Taken above, our study revealed that TTN-AS1 played an important role in LUAD progression. TTN-AS1/miR-142-5p/CDK5 regulatory axis may serve as a novel therapeutic target in the treatment of LUAD. Although great progress has been made in diagnosis and treatment recently, the long-term clinical prognosis and overall survival of lung adenocarcinoma (LUAD) patients remain unsatisfactory2. The inadequate comprehension of LUAD-related biological mechanisms limits the improvement in therapeutic efficacies. Thus, it is urgent to further elucidate the involved mechanisms of cancer progression to improve the clinical outcome of LUAD patients.Lung cancer is the most common malignant tumor, constituting the leading cause of cancer-related death worldwide. Non-small cell lung cancer (NSCLC) accounts for 85% of lung cancers, and adenocarcinoma is the predominant histologic subtype at present3. Benefiting from the advances in high throughput analysis, increasing cancer-related long non-coding RNAs (lncRNAs) have been identified4. LncRNAs are defined as a class of transcripts longer than 200 nucleotides that lack any detectable open reading frame5. Accumulative studies have proved that lncRNAs exert pivotal roles in the regulation of gene expression. Aberrant expression of various lncRNAs have been identified in different tumors, including colorectal cancer, gastric cancer, breast cancer, NSCLC, etc.9 lncRNAs could function as either oncogene or tumor suppressor gene to participate in the pathogenesis and development of tumors, and some lncRNAs might be potential biomarkers for predicting tumor invasion and metastasis11. Certain differentially expressed lncRNAs may function as prognostic biomarkers for cancer patients12. lncRNAs can regulate gene expression via multiple ways, such as chromatin modification, transcription, and posttranscriptional regulation13. Emerging studies indicated that lncRNAs were able to function as critical elements of the competing endogenous RNA (ceRNA) network via binding miRNAs to neutralize their suppressing effects on target genes14. Recently, numerous lncRNAs have been proved to indirectly regulate gene expression by functioning as ceRNAs in NSCLC progression15.Aberrant expressions of oncogene and tumor suppressor gene are frequently observed in NSCLC and account for the carcinogenesis20. Importantly, TTN-AS1 exerted regulating effects via functioning as a ceRNA to sponge different miRNAs20. For instance, TTN-AS1 facilitated the proliferation and metastasis ability of esophageal squamous cell carcinoma by the modulating miR-133b/FSCN1 axis16. To the best of our knowledge, the expression and significance of TTN-AS1 in LUAD has not been investigated so far. In the present study, we first identified an upregulated expression of TTN-AS1 in LUAD by analyzing the data in The Cancer Genome Atlas (TCGA) database. Then, we detected the expression of TTN-AS1 in LUAD tissues, and found upregulated expression of TTN-AS1 was positively correlated with advanced tumor stage and lymph node metastasis. Knockdown of TTN-AS1 significantly decreased the growth, proliferation, migration, and invasion ability of LUAD cells. In mechanism, we showed that TTN-AS1 could act as a ceRNA to bind to miR-142-5p and indirectly upregulate the expression of cyclin-dependent kinase 5 (CDK5) in LUAD. Herein, TTN-AS1 also exerted promoting effect on epithelial mesenchymal transition (EMT) process of LUAD cells. Taken together, this study unveiled a novel biomarker axis, consisting of TTN-AS1/miR-142-5p/CDK5, which might serve as a novel therapeutic target in treating LUAD.LncRNA titin-antisense RNA1 (TTN-AS1) is transcribed on the antisense strand of TTN and has partial sequence complementarity with TTN. TTN-AS1 has been identified to be an oncogene in numerous cancers, such as hepatocellular carcinoma, cervical cancer, papillary thyroid cancer, and gastric cancerP = 0.003). Then, we determined the expression of TTN-AS1 in 107 LUAD tissues and matched adjacent normal tissues to further confirm the data from TCGA database. Among the total 107 tumor tissues of LUAD patients, 58 cases (54.21%) showed high expression of TTN-AS1, compared with matched adjacent normal tissues. First, we analyzed the association between TTN-AS1 expression and clinical parameters. The results showed that the expression of TTN-AS1 was significantly correlated with the TNM stage and lymph node metastasis while not with gender, age, and invasion range. LUAD patients with higher TNM stage (stage III) and positive lymph node metastasis had significantly higher expression of TTN-AS1 than those with lower TNM stage (stages I and II) and negative lymph node metastasis (Table P < 0.001). Then the COX proportional hazard regression analysis was performed to further assess the predictive role of TTN-AS1 expression in LUAD prognosis. The covariates included in the COX proportional hazard model were gender, sex, invasion range, lymph node metastasis, TNM stage, and TTN-AS1 expression. After stepwise multivariate survival analysis, the TNM stage and TTN-AS1 expression were found to be significantly correlated with the DFS and OS . In addition, the results showed that the high TNM stage was associated with shorter DFS and OS, with the unadjusted hazard ratio (HR) being 3.130 (95%CI: 1.894–5.171) and 3.702 (95%CI: 2.146–6.389). As to high TTN-AS1 expression, the unadjusted HR being 7.870 (95%CI: 4.537–13.652), and 5.342 (95%CI: 2.937–9.717) for DFS and OS, respectively. The above results indicated that the high expression of TTN-AS1 might be involved with tumorigenesis and regarded as a potential predictor in LUAD patients.To determine the role of TTN-AS1 in LUAD progression, we first explored its expression by analyzing the data from TCGA database. The results demonstrated that TTN-AS1 expression was significantly upregulated in LUAD tissues, compared with normal tissues  and LncBase V2.0 (http://carolina.imis.athena-innovation.gr/diana_tools/web/)21 were adopted to predict potential miRNAs which might be sponged by TTN-AS1. As Fig. R2 = 0.335 and P < 0.001, Fig. P < 0.001; P = 0.006 and Fig. Since many lncRNA can exert various functions in cancers via binding to miRNAs, we then tried to explore the potential miRNAs which might be bound with TTN-AS1. Then, the webserver lnCeDB (http://www.targetscan.org/vert_71/) and miRanda (http://www.microrna.org) algorithms to predict potential target of miR-142-5p. CDK5 was predicted to be a target of miR-142-5p in lung. Using the Targetscan and miRanda algorithms, two hypothetic miR-142-5p binding sites were identified in the CDK5 3′-untranslated region  to determine the expression of CDK5 in LUAD specimens and analyzed its correlation with TTN-AS1. As Fig. 22, we then detected the effect of TTN-AS1/miR-142-5p/CDK5 axis on EMT-associated proteins. The results demonstrated that knockdown of TTN-AS1 led to an increase of E-cadherin and decrease of N-cadherin, Twist, Snail and zinc finger E-box binding homeobox (ZEB1) in A549 and H1650 cells, which could be reversed by addition of the miR-142-5p inhibitor , etc.24 lncRNAs are defined as a class of transcripts longer than 200 nucleotides that lack any detectable open reading frame5. Numerous lncRNAs have been confirmed to regulate the expression of target genes through multiple mechanisms25. miRNAs are evolutionarily conserved single-stranded RNAs containing about 21–24 nucleotides, which can trigger target mRNA degradation or translation inhibition by targeting its 3′-UTRs26. Binding to miRNAs to indirectly regulate the expression of target genes is one of the most frequent function mechanism of lncRNA15. Recently, a growing number of studies have elucidated that dysregulated lncRNAs play crucial roles in tumorigenesis of LUAD by participating multiple biological behaviors27. For example, the lncRNA linc00673 could bind to miR-150-5p to upregulate ZEB1 and promote EMT in NSCLC28. Overall, the interaction between lncRNA and miRNA plays pivotal roles in regulating malignant behaviors of cancers, thus further study on this interaction is beneficial for improving clinical therapeutic efficacy in treating LUAD. However, the function and mechanisms of many lncRNAs have not revealed in LUAD.LUAD still remains a malignant disease with unsatisfactory curative effects in the clinical practice. Activation of invasion and metastasis is one of the hallmarks of cancer, which is closely associated with poor prognosis of LUAD paitents18. Then, Cui et al. verified that TTN-AS1 promoted proliferation, migration, invasion, and EMT in PTC19. Lately, Dong et al. found that TTN-AS1 could promote the progression of gastric cancer via functioning as a ceRNA of miR-376b-3p to regulate KLF1220. Aiming at realizing the function of TTN-AS1 in LUAD, we first analyzed the expression of TTN-AS1 in LUAD with the help of TCGA database, the results showed that TTN-AS1 was a highly expressed lncRNA in LUAD. Then, to verify the results from TCGA database, we detected the expression of TTN-AS1 in LUAD tissues. Consequently, we found that TTN-AS1 was highly expressed in LUAD tissues comparing with matched adjacent normal tissues. In addition, high expression of TTN-AS1 in tumor tissues was significantly correlated with poor clinical parameters of LUAD patients. Meanwhile, survival analysis revealed that high TTN-AS1 expression was significantly associated with shorter DFS and OS, which suggested that TTN-AS1 might be a potential predictor for poor prognosis of LUAD patients. Aiming at further realizing the biological function of TTN-AS1, we then assayed its expression in LUAD cell lines, and found that A549 and H1650 cells showed the highest expression of LUAD. For realizing the role of TTN-AS1 in LUAD cells, we performed a knockdown of TTN-AS1 with shRNA. As a result, knockdown of TTN-AS1 significantly suppressed the growth, invasion, and migration ability of A549 and H1650 cells. Next, bioinformatics algorithms were used to explore potential binding miRNA of TTN-AS1 to determine its possible functional mechanism. Then, we chose miR-142-5p for our further study. miR-142-5p, identified as a tumor suppressor gene, is proved to be attenuated in various cancers such as osteosarcoma, ovarian cancer, gastric cancer, and NSCLC32. In the present study, we found that TTN-AS1 shared a complementary binding site with miR-142-5p, thus TTN-AS1 could bind to miR-142-5p and decrease its expression in LUAD cells. In addition, TTN-AS1 and miR-142-5p could be pulled down by anti-Ago2. Taken together, these data indicated that TTN-AS1 could act as a ceRNA to bind to miR-142-5p in LUAD cells.In the current study, we examined expression and biological functions of lncRNA TTN-AS1 in LUAD. TTN-AS1, a recently identified oncogene, has been proved to participate the progression of numerous cancers. Chen et al. showed that TTN-AS1 could upregulate E2F3 to promote growth and metastasis ability of cervical cancer via sponging miR-57335. Moreover, Sun et al. also found that CDK5 could induce EMT process in head and neck squamous cell carcinoma22. In the present study, we observed that the loss of miR-142-5p was positively correlated with CDK5 expression in LUAD tissues, and further confirmed that miR-142-5p could downregulate CDK5 in LUAD cell lines in vitro. Moreover, we found that the lncRNA TTN-AS1 could bind to miR-142-5p and increase the expression of CDK5 in LUAD cells. Since CDK5 has been reported to be correlated with EMT22, thus we next analyzed the correlation between EMT-related proteins and CDK5 in LUAD cells. EMT, a frequent malignant biological behavior, is the process by which epithelial cancer cells lose their cell–cell adhesion, and gain invasive and migratory properties to become mesenchymal stem cells28. The EMT program is important for NSCLC to invade their surrounding desmoplastic stroma36. E-cadherin is necessary for epithelial cells to maintain cell–cell adhesion, and the loss of E-cadherin is considered to be a fundamental event of EMT37. The activation of ZEB1 can downregulate E-cadherin to initiate the EMT process38. Then we detected the expression of EMT-related proteins in LUAD cells and found the knockdown of TTN-AS1 led to increased expression of E-cadherin and decreased expressions of N-cadherin, Twist, Snail and ZEB1. Meanwhile, addition of miR-142-5p inhibitor reversed this phenomenon, which confirmed the existence of TTN-AS1/miR-142-5p/CDK5 axis in LUAD. Thus, it could be seen that TTN-AS1 could participate the initiation of EMT through sponging miR-142-5p. As a result, suppression on the invasion and migration ability could be brought by knockdown of TTN-AS1 and rescued by miR-142-5p inhibitor in LUAD cells. Likewise, in the LUAD without knockdown of TTN-AS1, the miR-142-5p mimic could upregulate E-cadherin, and downregulate N-cadherin, Twist, Snail and ZEB1 to inhibit their invasion and migration ability. These results largely expanded the tumor suppressing effect of miR-142-5p and provided more supporting evidence for the ceRNA regulatory network.Aiming to further comprehending the function of TTN-AS1/miR-142-5p axis, we then predicted the potential target of miR-142-5p with bioinformatics algorithms. Then we found that miR-142-5p might target CDK5 by binding to its 3′-UTR region. CDK5, a member of CDK family, was considered to play an important role in numerous cancer-related biological behaviors such as cell proliferation, metastasis, angiogenesis, and immune escapeIn conclusion, To the best of our knowledge, despite of the studies on the role of TTN-AS1 in cancers, this is the first study to analyze its expression and function in LUAD. TTN-AS1 could sponge miR-142-5p to promote migration, invasion, and EMT ability of LUAD cells. High expression of TTN-AS1 was correlated with poor clinical parameters and postoperative prognosis of LUAD patients. Knockdown of TTN-AS1 could restore the suppressing effect of miR-142-5p on CDK5 expression, and led to inhibition of invasion, migration and EMT in LUAD cells. Meanwhile, these phenomena could be rescued by addition of miR-142-5p mimic. The present study revealed that TTN-AS1 might act as a ceRNA to indirectly regulate CDK5 by binding to miR-142-5p. Furthermore, the TTN-AS1/miR-142-5p/CDK5 axis might be used as potential biomarker in predicting prognosis of LUAD patients, and regarded as a novel target for anti-tumor therapies.The specimens of LUAD and matched adjacent normal tissues were collected from 107 patients who underwent surgery at the Fourth Hospital of Hebei Medical University  between Jan. 2014 and Jan. 2015. The median patient age at the time of surgery was 51 years (range: 29–73 years). None of the LUAD patients received preoperative treatment including radiotherapy, chemotherapy, or immunotherapy, etc. The clinical stage and histological tumor type were determined according to the UICC Classification of 2009 (seventh edition) and the WHO classification of 2005. Patients’ clinical information were collected and stored in a database, which was updated every 2 months by telephone follow-up. Complete follow-up was updated until death or Jan. 2019. The specimens were collected and treated promptly after surgery. The specimens were frozen and stored at −80 °C for extracting RNA. This research was approved by the ethic committee of the Hebei Medical University and all written informed consents were obtained before surgery.2 incubator.Human LUAD cell line H1650, HCC827, A549, H1975, PC9 and human bronchial epithelial cell line BEAS-2B were obtained from the research center of Peking Union Medical College . All cells were maintained in RPMI 1640  containing 5% FBS , 100 U/ml penicillin and 100 µg/ml phytomycin, at 37 °C in the humidified atmosphere of the 5% CO3 cells were plated into 96-well plates. When cells adhered, 10 μl of CCK8 (Solarbio) was added to each well and incubated for 2 h in a humidified incubator containing 5% CO2 at 37 °C. The absorbance of each well was detected at a wavelength of 450 nm. Proliferation rates were determined at 0, 24, 48, 72, 96 h after transfection. Experiments were performed in triplicate.The proliferation of cells was measured by the cell-counting kit-8 (CCK-8) assay according to the manufacturer’s protocol. Briefly, ~2 × 10Negative control (NC) shRNA and shRNA targeting TTN-AS1 were designed and cloned into lentivirus pCMV-VSV-G by Genepharma . The shRNA sequences were listed in Supporting Information Table −ΔΔCt 39. Human GAPDH and U6 snRNA were used for TTN-AS1 and miR-142-5p as internal control, respectively.In brief, total RNA was extracted from cells or tissue specimens with TRIzol reagent (Invitrogen) in accordance with the manufacturer’s instructions. Reverse transcription was performed using 1 μg of total RNA with SuperScript RT . For miRNA detection, the reverse transcription of miRNAs was with special primers . Proteins were separated by 10% SDS-PAGE and transferred electrophoretically onto polyvinylidene difluoride membranes . The membranes were incubated in PBS containing 5% bovine serum albumin for 2 h at room temperature, followed by overnight incubation at 4 °C with different dilutions of the primary antibodies, including antibodies to CDK5 , ZEB1 , E-cadherin , N-cadherin , Twist , Snail  and GAPDH . The membranes were developed with the Odyssey infrared imaging system according to the manufacturer’s instructions. The levels of protein in each sample were normalized relative to those of GAPDH. Each experiment contained triplicate wells of each sample, and all experiments were repeated at least three times.5 A549 or H1650 cells were seeded in 24-well plates. After scraping the cell monolayer with a sterile micropipette tip, the wells were washed three times with serum-free medium. The first image of each scratch was acquired at the time zero. After 24 h, each scratch was examined and captured at the same location, and the healed area was measured.A total of 5 × 102 humidified incubator at 37 °C. Cells on the upper side of the filters were removed by cotton-tipped swabs, and the filters were washed with PBS. Then the cells on the lower side were fixed in 4% formaldehyde and stained with 1% crystal violet in PBS for 5 min at room temperature. The cells on the lower side of the filters were defined as migration cells and counted at ×200 magnification in 5 random fields of each filter.Tumor cell migration assay was performed in a 24-well transwell chamber , which contained an 8 μm pore size polycarbonate membrane filter and was precoated with 100 μg Matrigel for invasion assay . Briefly, the cells were seeded in the upper chambers and incubated in 500 μl RPMI 1640 medium without FBS, while 500 μl medium with 10% FBS was placed in the lower chambers. The plates were incubated for 24 h in a 5% CO5 cells were resuspended in 100 μl 1× binding buffer, then 5 μl PI and 5 μl FITC Annexin V were added. After incubated for 10 min at room temperature in the dark, 400 μl 1× binding buffer was added to each tube and stained cells were analyzed by FACS Calibur Flow Cytometer (BD Biosciences).Apoptosis rate was assayed with FCM using the FITC Annexin V Apoptosis Detection Kit I  according to the manufacturer’s protocol. Briefly, the A549 or H1650 cells were harvested using trypsin without EDTA. After washed twice with precooling PBS, about 5 × 104 cells/well in 24-well plates and allowed to settle overnight. On the next day, the cells were transfected with recombinant plasmids or an empty plasmid encoding the firefly luciferase reporter with Lipofectamine 2000. Renilla luciferase reporter pRL-CMV  was cotransfected into cells as a normalizing transfection control. After 48 h, the reporter luciferase activity was measured with the Dual-luciferase Reporter assay system (Promega) according to the manufacturer’s instructions. All transfection assays were carried out in triplicate.Plasmids containing the firefly luciferase reporter were constructed with CDK5-3′-UTR-WT and CDK5-3′-UTR-MUT. Cells were seeded at 5 × 10RIP assays were performed using the Mana RIP RNA-Binding Protein Immunoprecipitation Kit (Millipore) according to the manufacturer’s instructions. In brief, A549 and H1650 cells were harvested and lysed with RIP lysis buffer containing protease inhibitor cocktail. Then the cell supernatants were incubated with magnetic beads conjugated with antibody against Argonaute 2  or IgG  for 2 h at 4 °C. Following this, RNase-free DNase I and Proteinase K were consecutively used to remove DNA and protein in the RIP complex. The gained RNA was subjected to qRT-PCR for detecting the enrichment of TTN-AS1 and miR-142-5p.40. The rabbit polyclonal antibody against CDK5  was used for detection. For evaluating expression of CDK5 in LUAD tissues, the staining was visualized and classified based on the percentage of positive cells and the intensity of staining according to the 0–4 semi-quantitative system described in our previous study40. The total scores were determined by multiplying the percentage score and intensity score and graded as low for score of 0–4 and high for score of 5–12. Each section was scored independently by two pathologists and a third pathologist determined the final score, if there was any inconsistency.IHC analysis was performed as we previously describedt test was used for analyzing the data with equal variance, and Mann–Whitney U test was used for those without equal variance. Spearman rank correlation was used to analyze the association between TTN-AS1 with miR-142-5p or between CDK5 with TTN-AS1 and miR-142-5p in LUAD tissue specimens. P value < 0.05 was considered as statistically significant, and all P values were two-tailed.Statistical analysis was performed by using SPSS statistics software, version 25.0 . All measurement data were presented as mean ± SD. The association between TTN-AS1 expression and clinical parameters was evaluated using the chi-square test or Fisher’s exact test. Survival analysis was carried out using the log-rank test in association with Kaplan–Meier analysis and Cox proportional hazards model analysis. Student Supplemental Table S1Supplemental Table S2


Although tobacco smoke has been associated with many infections, little is known of its association with human papillomavirus (HPV) infections among young adult women. The aim of the study was to explore the association of tobacco smoke exposure on HPV infections in young adult women in the United States. It was hypothesized that tobacco smoke exposure (both active and passive) as objectively measured by cotinine levels was associated with increased HPV infection in a national sample of 18 and 26 year-old women in the United States.The NHANES 2007–2012 dataset was used in the analyses. A national representative sample of women 18 to 26 year old  was included in the study. Infection with any HPV was determined by PCR while tobacco smoke exposure was determined by measuring serum cotinine levels. Women with cotinine levels <0.05 ng/mL were considered unexposed and those with levels > = 0.05 were considered as exposed. Exposed women were further categorized as passive smokers . Data were analyzed by descriptive statistics and multiple logistic regression analysis. Exposed women (passive smokers with cotinine levels > = 0.05ng/mL-10ng/mL) were almost 2 times (64% vs 35%) more likely to be infected with any HPV type than their unexposed counterparts (cotinine levels <0.05ng/mL). Also women who were active smokers (cotinine levels > = 10 ng/mL) were more than twice more likely (75%) to be infected with the virus than the unexposed counterparts. The relationship held true even after controlling for various socio-demographics. Indeed in the multiple regression analyses controlling for the various confounders, compared to their unexposed counterparts, women exposed to second hand smoke were more than twice more likely to have HPV infections . When compared to their unexposed counterparts, actively smoking women were more than 3.5 times more likely to be infected with HPV .th quartile .Finally, a strong dose-response relationship was further demonstrated with increasing risk of HPV with each quartile of cotinine levels even after controlling for various confounders. The risk of HPV was lowest in the lowest quartile (Referent OR = 1) and increased steadily with each quartile of cotinine levels until the risk was highest among women with cotinine levels in the 4Both passive and active tobacco smoking were strongly associated with any HPV infection in 18 to 26 year old young women with a significant dose-response relationship. Future studies should explore the effect of tobacco smoke exposure among younger women less than 18 years of age. Human papillomavirus (HPV) is probably the most common sexually transmitted infection in the United States affecting nearly all sexually active men and women. About 80 million Americans may be currently infected with the virus with almost 14 million new cases annually ,3.Known risk factors include, early initiation of sexual intercourse, frequency of sexual intercourse especially with multiple partners, ethnicity, and age –4. AlthoThe present study was aimed at determining the strength of the association of tobacco smoke exposure on HPV infections among a representative sample of young adult US women 18–26 years.The NHANES database was used for our analysis. The NHANES is a comprehensive research assessment of health and nutritional status of adults and children in the United States, which utilizes data collected through both interview and physical exam findings including medical, dental, and laboratory tests .NHANES data is and ongoing collection of data nationwide representative sample using stratified cluster complex sampling techniques by the CDC. Some population groups are over sampled in order to obtain accurate information on these groups not previously well studied. Full details of the complex sampling procedures have been described elsewhere .Subjects are questioned to obtain detailed socio-demographic information of all household members. In addition, subjects are asked to provide blood samples (in mobile examination centers (MEC), to assess serum cotinine levels. Serum cotinine levels were measured by an isotope dilution-high performance liquid chromatography . Data frTobacco smoke was classified into three levels. Subjects with serum cotinine levels <0.05 were defined as unexposed and non-smokers; individuals with levels of 0.05–10ng/mL were defined as exposed to second hand smoke, while those with levels > = 10 ng/mL were classified as active smokers. To further determine the effect of increasing cotinine levels on HPV infections, cotinine levels were categorized into 4 groups (quartiles).HPV infection was determine by the method of Roche Linear Array PCR using vaginal swabs and able to detect 37 HPV types viz 6,11,16,18,26,31,33,35,39,40,42,45,51,52,53,54,55,56,58,59,61,62,64,66,67,68,69,70,71,72,73,81,82,83,84,89,and 1539 [For the purpose of this analysis, race of the women was categorized as white and non-white.Average age of sex initiation was 16 years. The standard deviation of age at sex initiation was ~2 years. Therefore we dichotomized this variable into <14 years and 14+ years. The median number of lifetime sex partners was 4, with a mode of 1 so we dichotomized this variable into < 2 and 2 + for the analysis.The outcome variable of interest was HPV infection among 18 to 26year old women. The primary independent variable of interest was tobacco smoke exposure as objectively measured by cotinine levels.Four previously identified socio-demographic variables associated with HPV infections included to control for confounding were age of woman, race/ethnicity, life-time sexual partners, and age at sexual initiation. One potential confounder not previously explored was income.Descriptive statistics using X2 test for categorical variables were conducted to compare the proportions of subjects with HPV infections by selected socio-demographic variables. Multiple logistic regression analysis was further conducted to determine whether tobacco smoke exposure was still predictive of HPV infection after controlling for the other confounding variables. Because of the skewedness of cotinine levels, we further explored the impact of each quartile of cotinine level on HPV infection in order to determine if there was any dose-response relationship. A p-value of <0.05 was used as a test of significance for all cases. Because some population subgroups were oversampled to obtain unbiased estimates representative of US population, analyses were performed using the IBM SPSS complex sample analyses software for Windows, Version 240, Armonk, NY .th quartile of cotinine levels with a strong dose-response relationship (p <0.001).The other variables associated with HPV infections were annual household income (p = 0.021), race (p = 0.022), and age at sexual initiation (p = 0.030). Poorer women , while only 40% were unexposed Click here for additional data file.


G12D mice. Furthermore, α‐Mangostin inhibited the invasive and metastatic ability of pancreatic CSCs by suppressing the epithelial‐to‐mesenchymal transition (EMT) via up‐regulation of E‐cadherin and down‐regulation of mesenchymal phenotype by inhibiting N‐cadherin, Snail and Slug expression. Interestingly, the pluripotency maintaining factors and CSC markers were inhibited by α‐Mangostin thus suggesting that α‐Mangostin can target CSCs to inhibit pancreatic cancer effectively. Gli signalling plays a crucial role in the self‐renewal and pluripotency of CSCs. α‐Mangostin inhibited the Gli transcription and the expression of Gli target genes  in CSCs. Using ChIP assay, we demonstrated that Nanog could directly bind to promoters of Cdk2, Cdk6, FGF4, c‐Myc and α‐Mangostin inhibited Nanog binding to these promoters. Conversely, the inhibitory effects of the α‐Mangostin on CSC proliferation and Gli or Nanog transcription and their targets were abrogated by either enforced activation of sonic hedgehog (Shh) or by the overexpression of Nanog. Taken together, our studies suggest that α‐Mangostin may act as Gli inhibitor and establishes the pre‐clinical significance of α‐Mangostin for the prevention and treatment of pancreatic cancer.The current investigation was intended to elucidate the molecular mechanism of α‐Mangostin in the regulation of pancreatic cancer stem cell (CSC) characteristics. Here, we demonstrate that α‐Mangostin inhibited cell proliferation in pancreatic CSCs and cancer cell lines while it showed no effect on human pancreatic normal ductal epithelial cells. Also, α‐Mangostin inhibited colony formation and induced apoptosis in these cells. Further, α‐Mangostin inhibited the self‐renewal capacity of CSCs isolated from human primary tumours and Kras Mangosteen plant (Garcinia mangostana) grows abundantly in Southeast Asia mainly in the Sunda Islands and the Moluccas of Indonesia and other parts of tropical South America.G12D mice was inhibited by α‐Mangostin.The objective of this study was to delineate the molecular mechanism of α‐Mangostin in the regulation of pancreatic CSC characteristics. Our data demonstrate that α‐Mangostin inhibits cell proliferation in pancreatic CSCs and cancer cell lines. α‐Mangostin is non‐toxic to human pancreatic normal ductal epithelial cells. Also, α‐Mangostin inhibits the colony formation and induces apoptosis selectively in pancreatic CSCs and cancer cell lines, compared to the pancreatic normal ductal epithelial cells. Further, the self‐renewal capacity of CSCs isolated from human primary tumours and KrasFurthermore, α‐Mangostin inhibited the invasive and metastatic ability of pancreatic CSCs by modulation of cadherin expression and suppressing the transcription factors Snail, and Slug, which regulates the EMT. α‐Mangostin inhibited the expression of Nanog and pluripotency promoting factors  in CSCs. Using ChIP assay, we demonstrate that Nanog can directly bind to promoters of Cdk2, Cdk6, FGF4, c‐Myc and this Nanog binding was inhibited by α‐Mangostin. Interestingly, α‐Mangostin inhibits Gli transcription and expression. Conversely, the inhibitory effects of α‐Mangostin on pancreatic CSC proliferation and Gli or Nanog transcription and their targets was abrogated by either enforced activation of Shh or by the overexpression of Nanog. Taken together, these results suggest that α‐Mangostin may act as Gli inhibitor and also establishes the pre‐clinical significance of α‐Mangostin for the prevention and treatment of pancreatic cancer.22.1CD24, CD44, CD133, Nanog, Oct4, Sox2, KLF4, c‐Myc, Gli1, Gli2, Patched1, Patched2, Smoothened, Bcl‐2, Cyclin D1, E‐cadherin, N‐cadherin, Snail, Slug and Nanog antibodies were obtained from Cell Signaling Technology . Shh protein and anti‐β‐actin antibody were purchased from Santa Cruz Biotechnology Inc . α‐Mangostin (98% pure) was obtained from the LKT . Accutase was purchased from Innovative Cell Technologies, Inc . Matrigel was purchased from BD Bioscience . Crystal violet was purchased from Sigma‐Aldrich . TRIZOL was purchased from Invitrogen . Luciferase assay kit was purchased from Promega.2.22.Pancreatic cancer cell lines AsPC‐1 and PANC‐1 were used, and these cells were purchased from American Type Culture Collection  purchase. Cells were grown and frozen in liquid nitrogen for future use. Cells from second and third passages were used for the experiments. ATCC utilizes short tandem repeat (STR) profiling to authenticate the cell lines. PANC‐1 possesses mutations in p53 and K‐ras genes in codon 273 and codon 12 respectively. AsPC‐1 harbours mutation on codon 12 of K‐ras gene. CD133+/CD44+/CD24+/ESA+human pancreatic CSCs were isolated from primary tumours as described previously.2.34) in 1 mL of culture medium were incubated for 24 to 48 hours time‐points with various concentrations of 0‐10 µmol/L of α‐Mangostin. Cell viability and apoptosis were measured by trypan blue assay using Countess™ Automated Cell Counter (Invitrogen) and TUNEL assay respectively.Pancreatic cancer cells and CSCs . About 2 μg of the extracted RNA was reverse transcribed into cDNA using High‐Capacity cDNA Reverse Transcription Kit (Thermo Fisher Scientific). qRT‐PCR was carried out using fast SYBR Green Master Mix (Thermo Fisher Scientific). Relative mRNA expressions were compared with controls and evaluated using 22.8Cells were lysed in RIPA buffer and sonicated. The extracts were further centrifuged, and the supernatant was collected and stored at −80°C for future experiment and Western blot analysis. iMark (BIO‐RAD) protein assay was used to quantitate the cell lysates. 40‐50 μg of protein lysate was loaded on 10% or 12% bis‐acrylamide gels for protein separation and transferred onto PVDF membranes. The blot was blocked with 5% nonfat dry milk in TBS‐T (TBS and 0.01% Tween‐20) buffer and incubated with various primary antibodies in TBS‐T overnight at 4°C. The immunoblots were washed thrice with TBS‐T and incubated with secondary antibody (1:20 000). For chemiluminescence reactions, Super Signal West Pico substrate  was used as per the manufacturers' protocol. For reuse, the blots were washed in stripping buffer for 30‐60 minutes at room temperature.2.9Pancreatic CSCs were treated with various doses of α‐Mangostin, crosslinked and sonicated. The crosslinked sheared chromatin samples were then incubated with 3 μg of Nanog antibody and 5 μL of protein‐A and protein‐G magnetic beads. The ChIP DNA was further purified, and 1‐3 µL elutions were used to measure enrichment using quantitative real‐time PCR. ChIP‐derived DNA was electrophoresed on 2% agarose gels.2.10Measurement of Gli and Nanog reporter activities was performed as we described elsewhere.2.11t test or ANOVA was used to analyse the differences between groups. Differences among groups were considered significant at P < 0.05.The results presented are representative of three independent experiments run in triplicate, unless otherwise indicated. Student 33.1Unregulated growth of cancer cells is a hallmark of several solid tumours including pancreatic cancer. We, therefore, focused on examining the effects of α‐Mangostin on cell proliferation. Pancreatic cancer stem cells (CSCs), cell lines (AsPC‐1 and PANC‐1) and human pancreatic normal ductal epithelial cells (HPNE) were exposed to various concentrations of α‐Mangostin. As demonstrated in  significantly increased the Nanog expression as compared with that in CSCs/Empty vector group Figure D. Cell p4In the current study, we demonstrate that α‐Mangostin inhibits pancreatic CSC characteristics through Shh‐Nanog pathway. Specifically, α‐Mangostin inhibits markers of pancreatic CSCs, pluripotency maintaining factors and Nanog transcription. α‐Mangostin inhibits cell motility and markers of EMT in CSCs and may act as a Gli transcription inhibitor. The inhibitory effects of α‐Mangostin on pancreatic CSC proliferation and Gli or Nanog transcription and their targets were abrogated by either enforced activation of Shh or by the overexpression of Nanog.We and others have documented the biological significance of pancreatic CSCs. α‐Mangostin, a non‐toxic xanthonoid, inhibits the cell proliferation in pancreatic CSCs and cancer cell lines. Also, α‐Mangostin inhibits the colony formation and induces apoptosis in these cells as well, compared to the pancreatic normal ductal epithelial cells. Several recent studies have demonstrated the roles of CSCs in malignant transformation, cancer initiation, metastasis, recurrence and drug resistance. The ability of α‐Mangostin to inhibit pancreatic CSC markers CD24, CD44 and CD133 demonstrate that it can modulate the tumour growth by the suppression of the CSC population. Furthermore, the ability of α‐Mangostin to suppress the expression of pluripotency maintaining genes Oct4, Sox2, KLF4 and c‐Myc suggest that it can directly control cancer stem cells, proliferation, pluripotency and self‐renewal.Sonic hedgehog pathway in pancreatic cancer has been shown to be constitutively active and plays a significant role in the initiation, progression and metastasis.Nanog has been shown to play an essential role in the maintenance of self‐renewal capacity and pluripotency in embryonic stem cells and progenitor cells.Inhibition of Shh pathway at the level of Gli transcription will be more effective in treating patients because mutations in smoothened receptors (upstream of Gli) are commonly observed during treatment. Our studies define the new mechanism(s) and novel molecular targets of pancreatic cancer treatment and prevention. α‐Mangostin can regulate progression and metastasis through the inhibition of Shh signalling pathway and factors controlling pluripotency and EMT. In the present study, we have used α‐Mangostin doses up to 10 µmol/L, which are comparable to other chemodietary agents in the clinic. This dose is low enough to exert clinical impact and may not need any further modification. In another study, we have demonstrated the pre‐clinical potential of α‐Mangostin‐encapsulated PLGA nanoparticles for the management of pancreatic cancer.Both EMT and CSCs have been increasingly demonstrated to be associated with the development of invasive characteristic and distant metastasis.In conclusion, we demonstrate here that α‐Mangostin inhibits pancreatic CSC characteristics and cancer cell growth through the inhibition of Shh‐Gli pathway. α‐Mangostin inhibits Gli transcription, and its target genes Nanog, Oct4, c‐Myc, Sox2 and KLF4. α‐Mangostin not only inhibited smoothened but also Gli transcription. Therefore, α‐Mangostin has an additional clinical advantage, that is, if the pancreatic cancer patients develop resistance to smoothened inhibition due to mutation(s) on the smoothened receptors, they will be able to respond to α‐Mangostin because it also inhibits the expression of effector molecule Gli. Therefore, targeting the Shh‐Gli pathway through dual inhibition (smoothened and Gli) by α‐Mangostin could have enormous clinical significance for pancreatic cancer initiation, progression, metastasis and tumour growth. Overall, our study suggests that α‐Mangostin can be used for the treatment and prevention of pancreatic cancer by targeting CSCs.YM, W.Y and AS have declared that no competing interests exist. SS and RKS have declared intellectual property interest.WY and YM performed the experiments, analysed the data and wrote the manuscript; SS, AS and RKS designed the study and contributed reagents. All authors read and approved the manuscript.


The Medical Library Association (MLA) held its 118th annual meeting in Atlanta, Georgia, May 18–23, 2018, at the Hyatt Regency Atlanta. The meeting theme was “Adapting, Transforming, Leading.” Total attendance for the meeting was 1,670, with 236 participating in continuing education courses. Additional meeting content—including the meeting program and various electronic presentations from the business meeting, plenary sessions, poster sessions, and program sessions—can be accessed by all meeting registrants via the MLA ’18 website.Executive Director Kevin Baliozian welcomed attendees and gave a pre-session announcement about a suicide in the hotel in a public area the night before. He discussed the steps MLA was taking to reach out to members who had been present as well as counseling services provided by the hotel. He then introduced MLA President Barbara A. Epstein AHIP, FMLA.MLA President Barbara A. Epstein, AHIP, FMLA, welcomed attendees to the 2018 annual meeting. She updated the audience on a change to the meeting format in regard to how awards would be given out over a series of sessions, rather than all at once, and encouraged everyone to attend the plenary sessions and visit with vendors throughout the meeting. President Epstein then introduced Connie K. Machado, AHIP, who welcomed attendees on behalf of the Southern Chapter.Connie K. Machado, AHIP: Good morning and welcome. The Southern Chapter and all its members extend a warm welcome to you in beautiful downtown Atlanta on behalf of MLA for this 118th annual meeting. Many of our local Southern Chapter members have worked with MLA to plan this exciting meeting and venue for all of you in attendance. We hope that you are able to adapt to our humidity and temperatures while you’re here in the South.Founded in 1951, the Southern Regional Group later transformed to the Southern Chapter and has hosted three MLA meetings in this region. It has been seventeen years since the last meeting, which was held in Orlando, and we welcome you back with our warm Southern charm, great food, and hospitality. The Southern Chapter has a reputation for having fun and hosting great social events, so we hope you take some of our spirit of Southern hospitality home with you.Throughout the Southern Chapter’s rich history, many of our founding librarians have served as MLA president and in other positions as leaders. Names like Eileen Cunningham, Mildred Crow Langner, and Mary Louise Marshall, as well as William (Bill) Postel, Alfred N. Brandon, Mildred Jordan, T. Mark Hodges, and a host of others. And I’m sure that there will be more leaders to come in the future.Most of you have passed through Atlanta when traveling, at least at the airport. But there is much more to see and absorb here in the city’s rich history. Southern Chapter members are pleased to welcome you to Atlanta, home to the Centers for Disease Control and Prevention (CDC), Coca-Cola, and CNN, and plenty of dining experiences and fun places to visit during your stay. They also have seventy-five streets with the name “Peach” in it, so navigate carefully. So, again, I welcome you on behalf of the Southern Chapter members and MLA.Now, please join me in welcoming these truly creative and dedicated members who have spent more than three years planning this Atlanta meeting: the 2018 National Program Committee (NPC) Cochair David A. Nolfi, AHIP; 2018 NPC Cochair Debra Berlanstein, AHIP; 2018 Local Assistance Committee Chair Sandra G. Franklin, AHIP, FMLA; and 2018 Local Assistance Committee Cochair Joe Swanson Jr. Let’s give them a big hand.David A. Nolfi, AHIP: Thank you, Connie. On behalf of the MLA 2018 National Program Committee and the Local Assistance Committee, we officially welcome you to Atlanta, the first time for the MLA annual meeting. Our theme, “ATL: Adapting, Transforming, Leading,” reflects both this vibrant city and the spirit of our membership. The rising phoenix in our logo reflects the rise of Atlanta to become the dynamic leader in commerce, health care, research, education, and entertainment that it is today.Debra Berlanstein, AHIP: As medical librarians, we too, are continually evolving and searching for innovative ways to grow, change, reimagine, and most of all, make a difference. We know you will be inspired by the programming and the many opportunities to connect and share with colleagues during your stay in this amazing city. We hope you will get out and about and explore the world outside the doors of the Hyatt Regency Atlanta.David Nolfi: Planning this meeting over the last two years gave us the privilege of working with a great group of enthusiastic librarians whose hard work, energy, and commitment made cochairing the committee a pleasure. And so I ask now, would the members of the National Program Committee as well as the MLA staff who worked on the meeting please stand to be recognized. And now I ask you to please welcome Sandra Franklin, chair of the Local Assistance Committee, and Joe Swanson Jr., cochair.Candide, by Leonard Bernstein.Joe Swanson Jr.: Welcome to Atlanta. We’re so glad that you’re here and we hope you have an enjoyable time. We want you to enjoy the sessions and so forth. But in the meantime, we want you to have a little fun. We know that some of you have already been out and about, but you can visit places like the botanical gardens, the Jimmy Carter Library and Museum. You’ve already been around the attractions at the Centennial Olympic Park. And the Atlanta Symphony Orchestra will be performing Applause.]So, as I said, we want you to enjoy the meeting, but in your spare time and for those who are going beyond the meeting, enjoy the city. And come back to see us! We have what we call Southern hospitality here. Thank you.  They’re scattered hither and yon, but just thank them if they’re near you.In addition to that, I’d like to thank personally everyone who has worked on this committee, but especially our Local Assistance Committee. They each had their tasks, they did their tasks and just did a fantastic job, and you’re seeing the results of that. So I’d like all of them to stand. Please stand [and be recognized] for this 2018 meeting. , in building stronger relationships between these groups. And if you don’t know what the NIH All of Us research program is, plan to be here on Wednesday morning to hear Dr. Dara Richardson-Heron.This is an initiative of MLA and the Public Library Association, and it is funded by the National Network of Libraries of Medicine (NNLM) and the NIH All of Us research program. And this unique partnership allows us to leverage the expertise of our members, of you And third, my family. We have two great daughters who married two great sons-in-law, who brought us four terrific grandchildren. And the littlest one is Hartley, who was born just last month, and she’s still tiny. She’s eating a lot, she’s more than eight pounds now, and we expect her to grow into her hair bows very soon. [Applause.]And last but not least, my husband, Arnold. Through the years, he has been my confidante, my sounding board, my gentle critic, my press agent, my legal advisor, my best friend, and my secret sauce. I have dragged him to a few MLA meetings, and I have to say he sometimes enjoys them way more than I do. Last night, he came back from the exhibit opening with just a whole bagful of stuff that he collected and a list of people he talked to. And I think marrying him was the best decision I ever made. Thank you. If you have any questions, you’re welcome to email Cujo, my dog, and he will respond to you, or you can email me or Ray, whatever you prefer. Thank you for all of your time.  Why has it increased? Because of new programming, like the InSight Initiative, sponsored by the participating organizations, that allowed us to have the librarian attendees attend for free and pay for travel expenses, which is great.That number has increased by an impressive $46,000 this year, affecting 150 individuals. And lastly, after this meeting, vacation and kind of a zen in Canada on Lake Huron. And so between the graduation and that, I’ve had the pleasure of being here and meeting all of you. Thank you.  Now, I hope most of you know who Chef Ramsay [Gordon Ramsay of Hell’s Kitchen] is, because if you don’t, this won’t make any sense at all. And anyway, what does that have to do with libraries, you say? Well, nothing. But it does have an analogy to what I want to say.So, I’ve titled this time with you today: If Chef Ramsay was coming to your house, would you cook? So, I’m in the kitchen, but I’m not necessarily the expert. We, collectively, are the experts. And if Chef Ramsay was coming to my house, heck, no, I wouldn’t cook, because he scares me. But I’m not afraid of you, because you are all my colleagues, and I feel comfortable in saying this: In the 120th year of the association, I am proud and honored to be the first African American president of the Medical Library Association. We are changing. We are becoming a more diverse society. But our profession has not yet caught up to that. And I want to thank Elaine R. Martin, as she so beautifully articulated that yesterday in her Doe lecture, so thank you Elaine. You said everything I wanted to say and didn’t have time. So, over the next year, I want you to remain calm, I want you to remain open, I want you to remain flexible, and I want you to remain positive. As we’re sitting in this room here in Atlanta—the home of Coke, fried foods, grits, and pie—we’re making history together. Thanks for taking this journey with me and helping to shape the framework for the next year and years to come. You will always be the sunshine in my life. Thank you. [Applause.]I’d like to thank everyone. And please join us tonight for the Silver and Gold Networking Dinner, where we will present a few remaining awards and be entertained by our colleagues at MLA’s first-ever talent show. And get a sneak-peek at next year’s meeting, when the 2019 National Program Committee presents the MLA ’19 invitation. This session is now closed. [The Silver and Gold Networking Dinner, sponsored by McGraw-Hill Education, was held on Tuesday, May 22, 2018, from 6:30 p.m.–10:00 p.m. The networking dinner was in a different format than in previous years and featured a talent show, which included a live band, singing, a sing-along, and poetry. The MLA ’19 Invitation to Chicago also took place and was hosted by James Dale Prince, AHIP, chair, 2019 National Program Committee, and executive director, Southeastern/Atlantic Region, National Network of Libraries of Medicine, University of Maryland–Baltimore; Mellanye J. Lackey, cochair, 2019 National Program Committee, and associate director, Education and Research, Spencer S. Eccles Health Sciences Library, University of Utah–Salt Lake City; Rosie Hanneke, AHIP, chair, 2019 Local Assistance Committee, and information services and liaison librarian, Library of the Health Sciences, University of Illinois–Chicago; and Debra Werner, cochair, 2019 Local Assistance Committee, and director, Library Research in Medical Education, John Crerar Library, University of Chicago, Chicago, IL. The Dental Section, recipient of the MLA Section Project of the Year Award, and the Mid-Atlantic Chapter of MLA, recipient of the Majors/MLA Chapter Project of the Year (sponsored by J.A. Majors), were recognized during the dinner.Journal of the Medical Library Association.Section programs were presented in 4 time slots: Sunday, May 20, 3:00 p.m.–4:25 p.m.; Monday, May 21, 10:30 a.m.–11:55 a.m., and 1:00 p.m.–2:25 p.m.; and Tuesday, May 22, 3:00 p.m.–4:25 p.m. Paper abstracts that were scheduled to be presented are available on the MLA ’18 website. The final version of the abstracts, reflecting only those presented at the meeting, is included as an online-only Journal of the Medical Library Association.Poster sessions were presented in 4 time slots: Sunday, May 20, 2:00 p.m.–2:55 p.m.; Monday, May 21, 2:30 p.m.–3:25 p.m.; and Tuesday, May 22, 1:00 p.m.–1:55 p.m., and 2:00 p.m.–2:55 p.m. Poster abstracts that were scheduled to be presented are available on the MLA ’18 meeting website. The actual posters are available online in the MLA ’18 meeting website. The final version of the abstracts, reflecting only those posters presented at the meeting, is included as an online-only On Thursday, May 17, the MLA Board of Directors met. The MLA Board of Directors and Credentialing Committee met on Friday, May 18. On Saturday, May 19, the following groups met: 2019 National Program Committee; 2019 program planners; Chapter Council; Communities Strategic Goal Task Force; Joint Section Council/Chapter Council; National Network of Libraries of Medicine (NNLM) Steering Committee; Nominating Committee; and Section Council.JMLA Editorial Board; Leadership Curriculum Committee; Metadata 2020: Join the Discussion to Help Improve the Quality of Metadata for Research; Midcontinental Chapter Meeting and Greet #1; Outreach and Marketing SIG Business Meeting; Research and Evidence-Based Practice Curriculum Committee; Research Section Research Award Judging; Research4Life Grants Workshop Planning Session; Resource Sharing SIG Business Meeting; and Systematic Reviews SIG Business Meeting. Exhibitor meetings included: Lunch & Learn: AAAS/Science: The Blurred Line between Fact and Fiction; Medical Librarian Lunch and Learn: EBSCO Health; Cell Press & The Lancet Lunch & Learn: Best Practices in Publishing and Reproducibility; Supporting Institutional Research and Raising the Profile of the Library; and The R2 Digital Library as a Health Sciences eBook Database.On Sunday, May 20, the following groups, sections, and SIGs met: AAHSL Future Leadership Committee; AAHSL Leadership Fellows Program; AAHSL New and Developing Libraries Committee; Ad Hoc Committee to Review Core Clinical Journals; African American Medical Library Alliance SIG Business Meeting; chapter treasurers orientation meeting; Collection Development Section Business Meeting; Consumer and Patient Health Information Section (CAPHIS) Business Meeting and Executive Committee Meeting; Data SIG Business Meeting; Education Steering Committee; Educational Media and Technologies Section (EMTS) Business Meeting #1; Fellows of MLA; Health Disparities SIG Lunch; International Cooperation Section Business Meeting; Interprofessional Education SIG Business Meeting; On Monday, May 21, the following groups, sections, and SIGs met: 2020 National Program Committee; AAHSL Research Services Committee; Around the World: Global Librarians’ Experiences; Awards Endowment Task Force Meeting; Awards Committee; Books Panel; Cancer Librarians Section Business Meeting; Clinical Librarians and Evidence-Based Healthcare SIG Business Meeting; Complementary and Alternative Medicine SIG Business Meeting; Data Catalog Collaboration Information Session; Dental Section Business Meeting; Department of Veterans Affairs Librarians SIG Business Meeting #2; EMTS Business Meeting #2; Gaming in Adult Learning SIG Business Meeting; Government Relations Committee; Health Information Professionalism Committee; History of the Health Sciences Section Business Meeting; Hospital Libraries Section (HLS) Executive Board Meeting and Business Meeting and Social; Information Services Curriculum Committee; Instruction and Instructional Design Committee; Latino SIG Business Meeting; LGBQT Health Sciences Librarians SIG Business Meeting; Leadership Curriculum Committee; Libraries in Curriculum SIG Business Meeting; Medical Humanities SIG Business Meeting; Medical Informatics Section Business Meeting; Medical Libraries: Starting from Scratch; MLA Research Training Institute; Molecular Biology and Genomics SIG Business Meeting; New York-New Jersey Chapter Board Meeting; Nursing and Allied Health Resources Section (NAHRS) Business Meeting and Executive Board; Osteopathic Libraries SIG Business Meeting; Pacific Northwest Chapter Business Meeting; Pediatric Librarians SIG Business Meeting; Pharmacy and Drug Information (PDI) Section Business Meeting; Professional Recruitment and Retention Committee; Public Health/Health Administration Section Business Meeting; Public Services Section Business Meeting; Scholarly Communications Committee; Solo Librarians SIG Business Meeting; Technical Services Section Business Meeting; Transitional Sciences Collaboration SIG Business Meeting; Veterinary Medical Libraries Section Business Meeting; and Vision Section SIG Business Meeting. Exhibitor meetings included: Lunch & Learn: How to Conduct Systematic Reviews the JBI Way; and How to Provide a World-Class Systematic Review Service Using Covidence.On Tuesday, May 22, the following groups, sections, and SIGs met: AAHSL Program and Education Committee; Bylaws Committee; Diversity and Inclusion Task Force; education committee chairs joint meeting; Federal Libraries Section Business Meeting; Health Association and Corporate Libraries Section (HACLS) Business Meeting; Information Literacy in Medical Education (ILME) SIG Business Meeting; Information Management Curriculum Committee; Institutional Animal Care and Use SIG Open Discussion; Joint 2018 and 2019 Contributed Content Work Group; Leadership and Management Section Business Meeting; Joseph Leiter NLM/MLA Lectureship Committee; Librarians without Borders® Committee; Medical Library Education Section (MLES) Business Meeting; Medical Library Group of Northern California and Nevada Business Meeting; Medical Library Group of Southern California and Arizona Business Meeting; Membership Committee; Midcontinental Chapter Meeting and Greet #2; MLA community managers and webmasters; MLA News Editorial Board; Research Section Business Meeting; Rising Stars presentations; Rising Stars program; section treasurers orientation; Social Justice Section Business Meeting; Southern Chapter Executive Committee; Systematic Reviews SIG Informal Meeting; and Technical Services Section Social. Exhibitor meetings included: Lunch & Learn: Elsevier Luncheon for Medical Librarians: Collaboration for Innovation.On Wednesday, May 23, the following groups, sections, and SIGs met: Community Strategic Goal Task Force Meeting 2; Education Annual Programming Committee (EAPC); Grants and Scholarships Committee; and Oral History Committee.Three open forums were held on Sunday, May 20, from 4:30 p.m.–5:25 p.m.:MLA Communities Strategic GoalMLA Diversity and Inclusion Strategic GoalMLA InSight Initiative Summit 1 OutcomesThe National Library of Medicine (NLM) Update took place on Tuesday, May 22, 11:00 a.m.–11:55 a.m. Joyce E. B. Backus, associate director for library operations, began the session by introducing herself, Patricia F. Brennan, director of NLM, and Amanda J. Wilson, head, National Network Coordinating Office, National Network of Libraries of Medicine (NNLM), all of whom participated in the update.Dr. Brennan presented an explanation of the NLM Strategic Plan, “A Platform for Biomedical Discovery and Data-Powered Health 2017–2027.” The three pillars  were designed to target the future: (1) accelerate discovery and advance health through data-driven research; (2) reach more people through enhancement, dissemination, and engagement; and (3) build a workforce for data-driven research and health. In discussing the budget, Dr. Brennan said that NLM has received $22,000,000 for new investments in data science, accelerated training for librarians, and platform stabilization. She noted key accomplishments since February 2018.Ms. Backus reported on strategic planning activities and outcomes. She discussed MEDLINE 2022, a 5-year plan to do a behind-the-scenes modernization of MEDLINE and more efficient delivery of information on TOXNET. She introduced PubMed Labs and PubOne that launched in October 2017 and is a test site to try new features for PubMed 2.0, which will be released in December 2018. The NLM website has been refreshed: the PubMed Health Portal will be going away, but PubMed and Bookshelf will still have systematic review and drug information. MedlinePlus now includes Lab Test information. Ms. Backus mentioned several other NLM activities including the DeBakey Fellowship, rare book loans, visiting library and information science students, and NLM associate fellows.JAMA article on the history of NNLM from 1985–2015.Ms. Wilson noted NNLM year three activities and illustrated an organizational chart of NNLM. She highlighted projects in health literacy, hurricane relief, funding for classes, and the Wikipedia edit-a-thon. She showed videos of three examples of NNLM activities: University of Arizona, St. Paul, MN, and University of Pittsburgh Medical Center (UPMC). The All of Us public library initiative was also mentioned. Ms. Wilson showed a slide of the The remainder of the session was devoted to questions and answers.The Legislative Update was held on Tuesday, May 22, from 1:00 p.m.–1:55 p.m. Cristina Pope, AHIP, chair, MLA Governmental, Relations Committee, and director, Health Sciences Library, State University of New York (SUNY) Upstate Medical, University–Syracuse, moderated the session. Dina Paltoo, interim assistant director for policy development at NLM, presented information on the following: an overview of funding, functions of NLM, NLM’s most heavily used databases, statistics on PubMed Central, statistics on biomedical informatics and data science research, legislation and NLM, and bills and government activities that most affect NLM.Welcome Reception and Opening of the Hall of Exhibits, 5:00 p.m.–7:30 p.m.MLA New Members/First-Time Attendees Program and Breakfast, 7:00 a.m.–8:55 a.m.Yoga Class, 7:30 a.m.–8:30 a.m.Library School Reunion Tea, 2:00 p.m.–2:55 p.m.Clinical Librarianship Happy Hour, 5:30 p.m.–7:00 p.m.International Visitors Reception, 6:00 p.m.–7:00 p.m.Academy of Health Information Professionals Q&A Session, 2:30 p.m.–3:25 p.m.Diversity and Inclusion Task Force Fish Bowl, 5:00 p.m.–6:25 p.m.Public Librarian Welcome Breakfast and Speed-Networking Event, 7:00 a.m.–8:55 a.m.Public Librarians Symposium, Overview and Sessions 1 & 2, 9:00 a.m.–10:25 a.m.Public Librarians Symposium, Morning Concurrent Sessions 1 & 2, 11:00 a.m.–11:55 a.m.Public Librarians Symposium, Afternoon Concurrent Sessions 1 & 2, 1:00 p.m.–1:55 p.m.MLA Book Authors and Prospective Authors Gathering, 2:00 p.m.–2:55 p.m.Public Librarians Symposium, Health Literacy Heroes Sessions 1 & 2, 4:30 p.m.–5:25 p.m.Silver and Gold Networking Dinner, 6:30 p.m.–10:00 p.m.Exhibitors held Sunrise Seminars to provide information and to introduce new products and services. The following seminars were held:Artificial Intelligence (AI) in Health Care: From Adoption to DisruptionCochrane “Inside/Outside”: Update from Carol LefebvreEmbase Seminar: Librarian Roles in Teaching Evidence-Based MedicineThe Power of Single Sign-On and the Benefits to AllAmerican Psychological AssociationSpringer Nature Competitive Intelligence: Research Trends in Health CareThree Trends That Eclipse Point-of-Care ToolsTen Technology Showcases were held throughout Sunday, Monday, and Tuesday:Automating the Systematic Review Process: An Overview of DistillerSR’s New Productivity EnhancementsCase Files: Teaching Case CollectionCovering Your Nursing Bases: Research, Decision Support, and CompetenciesSupporting Your Interdisciplinary Care Dream TeamGetting Beyond the Impact Factor: Using Bibliometrics for Research Evaluation at the NIH LibraryTaking Medical Research to the Next Level: 72,987,390 Searches Can’t Be WrongThe Ever-Evolving World of Medical E-BooksFrom Data Standards to DermExpert: The Story of How an Image Collection Learned to ThinkLooking to the Future with EndNoteTop Questions Medical Libraries Ask OpenAthensThe 2017/2018 Education Annual Programming Committee offered the following courses to 236 attendees on May 18 and 19, 2018.Instructors: Julie Glanville, MCLIP, associate director, York Health Economics Consortium, University of York, York, United Kingdom, and Carol Lefebvre, HonFCLIP, independent information consultant, Lefebvre Associates, Oxford, United KingdomCE100 Advanced Searching Techniques and Advanced Strategy Design, Instructors: Karen Gutzman, impact and evaluation librarian, and Patricia L. Smith, impact and dissemination librarian, Galter Health Sciences Library, Feinberg School of Medicine, Northwestern University, Chicago, ILCE200 Dissemination in Action: Communicating Research in a Digital World, Instructors: Sarah Young, senior librarian, Mellon Institute Library, Carnegie Mellon University, Pittsburgh, PA; Jin Wu, emerging technologies librarian, Norris Medical Library, University of Southern California–Los Angeles; and Ayaba Logan, research and education informationist and assistant professor, Medical Libraries, University of South Carolina–CharlestonCE502 Statistics 101: A Primer in Statistical Methods for Health Sciences Librarians, Instructors: Gwen Wilson, health sciences librarian, Mabee Library, Washburn University, Topeka, KS, and Kristen DeSanto, AHIP, clinical librarian, Health Sciences Library, Anschutz Medical Campus, University of Colorado–AuroraCE600 Building Partnerships with Faculty, Clinicians, and Other Stakeholders, Instructors: Erin Eldermire, head, Flower-Sprecher Veterinary Library, Cornell University Library, Ithaca, NY; Sarah Young, senior librarian, Mellon Institute Library, Carnegie Mellon University, Pittsburgh, PA; and Lenny Rhine, FMLA, coordinator, E-Library Training Initiative, Librarians without Borders®/MLA, Gainesville, FLCE101 Performing Systematic Reviews in Resource-Limited Settings, Instructors: Julie Glanville, MCLIP, associate director, York Health Economics Consortium, University of York, York, United Kingdom, and Carol Lefebvre, HonFCLIP, independent information consultant, Lefebvre Associates, Oxford, United KingdomCE102 Trials without Tribulations: Identifying Clinical Trials for Systematic Reviews and Other Clinical and Research Questions, Instructors: Wichor M. Bramer, biomedical information specialist, Medical Library, Erasmus University Medical Center, Rotterdam, the Netherlands, and Melissa L. Rethlefsen, AHIP, deputy director, Spencer S. Eccles Health Sciences Library, University of Utah–Salt Lake CityCE103 Effectiveness and Efficiency in Exhaustive Searches, Instructors: Rachel Pinotti, AHIP, assistant library director, Education and Research Services, Levy Library, Icahn School of Medicine at Mount Sinai, New York, NY; Antonio P. DeRosa, AHIP, oncology consumer health librarian and assistant librarian faculty, Samuel J. Wood Library and C.V. Starr Biomedical Information Center, Weill Cornell Medicine, New York, NY; and Diana Delgado, AHIP, associate director, User Support, Research and Education, Samuel J. Wood Library and C.V. Starr Biomedical Information Center, Weill Cornell Medicine, New York, NYCE104 Upping Your Reference Game: Technologies and Strategies for Value-Added Reference Services, Instructor: Michele Malloy, research librarian, Congressional Research Service, Library of Congress, Washington, DCCE105 Health Policy Research: Navigating Governmental and Legislative Sources, Instructors: Julia M. Esparza, AHIP, head, User Education and Outreach Services; Montie’ L. Dobbins, head, User Access Services/Circulation; and David C. Duggar, AHIP, head, Library Liaison Program; Health Sciences Library, Louisiana State University (LSU) Health–Shreveport; and Alexandria (Lexi) Brackett, AHIP, clinical support librarian, Harvey Cushing/John Hay Whitney Medical Library, Yale University, New Haven, CTCE300 Not Just Numbers: Teaching Students to Think Using Practical Curriculum Exercises, Instructor: Jessica Sender, librarian, College of Nursing, Michigan State University–East LansingCE301 Innovations in Nursing Information Literacy: New Technologies, Approaches, and Ideas, Instructors: Andy Hickner, web services librarian, Harvey Cushing/John Hay Whitney Medical Library, Yale University, New Haven, CT, and Susanna Galbraith, virtual services librarian, Health Sciences Library, McMaster University, Hamilton, ON, CanadaCE302 Peeking Under the Hood: Understanding, Assessing, and Improving Your Library’s Website and LibGuides, Instructors: Jamie Conklin, research and education librarian, and Megan von Isenburg, AHIP, associate dean, Library Services and Archives, Medical Center Library & Archives, Duke University, Durham, NCCE303 Evidence-Based Teaching: Finding and Applying the Best Evidence to Instruction, Instructors: Heidi Heilemann, AHIP, associate dean and knowledge management director, Lane Medical Library and Knowledge Management Center, Stanford University, Stanford, CA; M.J. Tooey, AHIP, FMLA, associate vice president, Academic Affairs, and executive director, Health Sciences and Human Services Library, University of Maryland–Baltimore; and Gabriel Rios, director, Ruth Lilly Medical Library, Indiana University–IndianapolisCE400 Do You Want to Be a Library Director? Knowledge, Skills, and Career Paths for Library Leaders, Instructor: Erinn Aspinall, AHIP, strategic initiatives librarian and communications coordinator, Bio-Medical Library, Health Sciences Libraries, University of Minnesota–MinneapolisCE401 Digital Storytelling: Communication for Greater Impact, Instructor: Ayaba Logan, research and education informationist and assistant professor, Medical Libraries, Medical University of South Carolina–CharlestonCE500 What Did You Hear? Qualitative Data Analysis, Instructors: Marci Brandenburg, bioinformationist, and Jean Song, assistant director, Academic and Clinical Engagement, Taubman Health Sciences Library, University of Michigan–Ann ArborCE501 Introduction to Visualization Principles, Instructor: Lisa Federer, AHIP, research data informationist, NIH Library, National Institutes of Health, Bethesda, MDCE503 Introduction to Data Analysis and Visualization with R, Instructors: David A. Nolfi, AHIP, health sciences librarian and library assessment coordinator, Gumberg Library, Duquesne University, Pittsburgh, PA, and Carolyn Schubert, interim director, Research and Education Services, and health sciences and nursing librarian, Carrier Library, James Madison University, Harrisonburg, VACE601 Learning to Liaise with Health Professions, The online itinerary planner (sponsored by Wolters Kluwer) allowed attendees to peruse programs and events online. Complimentary WiFi was available throughout the Hyatt Regency Atlanta courtesy of The JAMA Network. Live streaming was available on Twitter using the hashtag #MLANET18, and volunteer bloggers, the Local Assistance Committee, and the 2018 National Program Committee contributed to the official meeting blog with meeting tips, announcements, and more. For those seeking new jobs and prospective employers, the Job Placement Center was open from Saturday through Tuesday, and the MLA Resume Clinic was available Saturday through Monday. The Hall of Exhibits was open Saturday evening through Monday.AppendixClick here for additional data file.AppendixClick here for additional data file.


The Veterans Benefit Administration’s (VBA) Aid and Attendance enhanced pension benefit (A&A) is available to older, low-income Veterans who require assistance meeting their daily needs. However, reports indicate that A&A is underutilized with only 1/3 of eligible Veterans receiving this benefit. The objective of this mixed methods study is to characterize the variability in A&A enrollment across VA Medical Centers (VAMCs) and determine factors attributable to the variation. Using VA administrative data, we calculated the rate of enrollment in A&A among Veterans receiving pension. We then purposefully sampled 16 Chiefs of Social Work at VAMCs with the highest (n=7) and lowest (n=9) rates of enrollment. Interviews were transcribed, coded, and analyzed using conventional qualitative research methods. The rate of enrollment in A&A varies from <1% to 23% across VAMCs. VAMCs that had higher rates of enrollment were larger and more likely to be located in the South and Mid-Atlantic regions. Respondents at sites with low rates of enrollment indicate that education around the eligibility criteria is needed for VAMC staff. They also report that outreach to Veterans about this benefit is limited. Respondents at VAMCs with high rates of enrollment indicate that the relationships with VBA and Veterans Service Organizations facilitates access. Universally, respondents viewed the A&A benefit positively and note that it helps meet Veterans’ long-term care needs. As the Veteran population continues to age, it is important that VA ensure equal access to A&A for eligible Veterans. Implications of these findings and next steps will be discussed.


Personalized, precision, and risk-based medicine are becoming increasingly important in medicine. These involve the use of information about the prognosis of a patient, to make individualized treatment decisions. This has led to an accumulating amount of literature available on prognosis studies. To summarize and evaluate this information overload, high-quality systematic reviews are essential, additionally helping us to facilitate interpretation and usability of prognosis study findings and to identify gaps in literature. Four types of prognosis studies can be identified: overall prognosis, prognostic factors, prognostic models, and predictors of treatment effect. Methodologists have focussed on developing methods and tools for every step of a systematic review for reviews of all four types of prognosis studies, from formulating the review question and writing a protocol to searching for studies, assessing risk of bias, meta-analysing results, and interpretation of results. The growing attention for prognosis research has led to the introduction of the Cochrane Prognosis Methods Group (PMG). Since 2016, reviews of prognosis studies are formally implemented within Cochrane. With these recent methodological developments and tools, and the implementation within Cochrane, it becomes increasingly feasible to perform high-quality reviews of prognosis studies that will have an impact on clinical practice. Clinical practice roughly consists of diagnosis, prognosis, and treatment. Treatment selection used to be driven mostly by the diagnosis that was made, but prognosis has become increasingly important to inform evidence-based decisions about healthcare. Clinical trials mainly focused on estimating a single overall effect, while decisions are being made on an individual level. Some patients benefit more (or less) than average; therefore, identifying those subgroups with different effects of an intervention has become very important to guide evidence-informed decision making. This personalized, precision, and risk-based medicine all involves the use of prognostic and predictive characteristics to make individualized treatment decisions for patients. The shift to personalized medicine has led to an accumulating amount of evidence available from prognosis studies. Reviews of prognosis studies make this information overload informative and usable. They summarize and evaluate the available evidence and guide the interpretation of results, in order to facilitate optimal use of all existing evidence. Reviews of high quality provide trusted evidence for stakeholders, like clinicians and guideline developers, to help them deciding which prognostic model or factor to use in clinical practice or implement in evidence-based guidelines. In addition, reviews identify gaps and redundant or unnecessary studies in the scientific literature, highlight flaws in conduct and reporting of primary studies, and identify and indicate which further studies are needed –5. TherePrimary prognosis studies are presented as four types in the PROGnosis RESearch Strategy (PROGRESS) partnership series –9: overaAll types of primary prognosis studies can be summarized, evaluated, and interpreted in different types of systematic reviews, following the broad range of aims and objectives of the included prognosis studies. Reviews are, for example, helpful to give an overview of all available prognostic factors or models , to study the prognostic value of a certain externally validated prognostic model (e.g. the predictive (prognostic) performance of the Revised Cardiac Risk Score for cardiac outcomes after noncardiac surgery ), or theMethodological guidance for most steps of conducting a systematic review is currently developed. Amongst others, to facilitate searching and reduce the number of references to be screened, methodological search filters are available –18, and The growing attention for prognosis research and the increasing emphasis on the importance of prognostic information in clinical practice have led to the introduction of the Cochrane Prognosis Methods Group (PMG) in 2007 . Over thIn summary, systematic reviews are urgently needed to summarize the growing amount of prognostic evidence, to evaluate the available evidence and guide the interpretation of results, in order to facilitate optimal use of existing evidence for medical practice and policy making. With the recent methodological developments and tools for systematic reviews of prognosis studies, it becomes increasingly feasible to perform these reviews. With the implementation within Cochrane, it is ensured that reviews of high quality will be produced that will have an impact on clinical practice.


The indicating FTA card is a dry medium used for collection of cervical samples. HPVIR is a multiplex real-time PCR test that detects 12 high-risk human papillomavirus types (hrHPV) and provides single genotype information for HPV16, − 31, − 35, − 39, − 51, − 56, and − 59 and pooled type information for HPV18/45 and HPV33/52/58. The aim of this study was to evaluate whether a strategy with cervical samples collected on the FTA card and subsequently analysed with the HPVIR test complies with the criteria of the international guidelines for a clinically validated method for cervical screening.® HPV test) based on liquid-based cytology (LBC) samples. Two clinical samples (LBC and FTA) were collected from 896 participants in population-based screening. For evaluation of the specificity we used 799 women without ≥ CIN2, and for clinical sensitivity we used 67 women with histologically confirmed ≥ CIN2. The reproducibility was studied by performing inter- and intra-laboratory tests of 558 additional clinical samples.We performed a non-inferiority test comparing the clinical sensitivity and specificity of the candidate test (FTA card and HPVIR) with a clinically validated reference test . Adequate agreement of > 87% was seen in both the intra- and inter-laboratory comparisons.The clinical sensitivity and specificity for samples collected on the FTA card and analysed using the HPVIR test were non-inferior to samples analysed with the Cobas® HPV test based on LBC samples (non-inferiority test score, Samples collected on the indicating FTA card and analysed with HPVIR test fulfil the requirements of the international guidelines and can therefore be used in primary cervical cancer screening. In this® HPV test as reference test.One advantage of using the FTA card is their suitability for self-sampling, which represent an important development of cervical cancer screening. Recently, we have shown that repeated self-sampling on FTA cards is as reliable for HPV testing as assisted sampling by a midwife or gynaecologist , 15, andFor this study we invited 896 women who were scheduled for cervical cancer screening in Uppsala County during 2017 and 2018. Among these, 819 women visited a midwife surgery and performed a primary screening test with both methods. Seven hundred ninety-nine women were either HPV negative or had histologically confirmed < CIN2 and were included in the comparison between the candidate test and the reference test to assess the clinical specificity. To determine the sensitivity, we studied a cohort of 75 women scheduled for a colposcopy due to abnormal cytology or/and a positive Cobas® HPV test in primary screening. Of these, 67 women with histologically confirmed ≥ CIN2 were included in the analysis of sensitivity. The inclusion criteria for all participants in the study were ≥ 30 years of age and not hysterectomized, or pregnant.The included women either had a midwife or a gynaecologist collecting two cervical samples per participant using one cytobrush for each collection. One sample was applied to an indicating FTA elute micro card™  and the other sample was transferred to the liquid based medium (LBC) . Sampling was randomized, meaning that the first sample was allocated randomly to either the FTA card or the liquid based medium. The FTA card sample was analysed using the HPVIR test (candidate test) and the LBC sample using Cobas® HPV test (reference test) on the 4800 platform . Women with at least one HPV positive outcome of the tests were referred for colposcopy and biopsy. The study was approved by the regional ethics committee in Uppsala, Sweden (Dnr 2017/088).The inter-laboratory agreement and reproducibility of the candidate test (FTA card and HPVIR test) was determined using samples from 558 women, of which 284 (51%) had previously tested HPV positive using the Hybrid Capture 2 (HC2) test. These samples were originated from another cohort and collected in the Nelson Mandela Hospital (WSU) or Mbekweni Community Clinic, Eastern Cape, South Africa. One sample was collected on the FTA card and the other in a tube of liquid medium for HC2. The laboratory at University of Cape Town (UCT) performed the evaluation of the inter-laboratory agreement. The intra-laboratory reproducibility was studied by comparison of the results of two different research engineers (Operators) in Uppsala, processing the same set of 558 samples.Homo sapiens hydroxymethylbilane synthase; GenBank accession no. M95623.1) as a control that the sample contains enough cellular material for the test to be informative. The test also detects and quantifies HPV16, 18, 31, 33, 35, 39, 45, 51, 52, 56, 58 and 59, and provides single genotype information for all types except HPV18/45 and HPV33/52/58, which are detected together as two groups. The limit of detection (LOD) for HPVIR is 10 HPV copies per PCR. In order for a sample to contain sufficient amount of material for the HPV test to be informative, a threshold of 10 copies of the nuclear single copy gene per PCR is used [FTA cards were processed using an automated laboratory system  where a robot arm picked up each FTA card, took a photograph of the sample deposition area, and then calculated which part of the card that contained the highest concentration of cells using a machine learning software. The robot first collected 5 punches from a clean card area and thereafter it punched 4 circular pieces of 3 mm diameter from the sample deposit area and collected all 4 punches in a single well in a 96-well microtiter plate. DNA was then extracted as described earlier and the HPV test was performed using the real-time PCR assay HPVIR , 19. The® 4800 is a fully automated system for sample preparation and real-time PCR, including the FDA approved Cobas® HPV test. The Cobas® HPV test include 14 hrHPV  of which HPV16 and 18 are detected as single genotypes while the others types are reported as a group denoted ‘Other HPV’ types (12 pooled types). To perform this assay, the 20 mL tube with cervical sample in LBC were loaded into the Cobas® 480 instrument, and 400 μL from each tube was transferred to the extraction plate, after which the samples were lysed in the presence of a chaotropic reagent. The DNA was then purified by adsorption to magnetic glass particles, washed and eluted in 100 μL dH2O. Fifty microliter of this was mixed with PCR reagents for amplification in the Cobas® 480 instrument. All Cobas® tests were performed by an accredited laboratory .Samples were collected in 20 mL LBC medium and stored in room temperature until analysed. The CobasSamples for the Hybrid Capture® 2 assay (HC2)  were collected and handled according manufactures recommendations by the lab at University of Cape Town (UCT). HC2 is based on hybridization with RNA probes to detect 13 hrHPV . A cut-off value of RLU/CO = 1, is equivalent to 1 pg HPV DNA per mL sampling buffer was used as a threshold for an HPV positive test.® HPV genotyping test . To this end, 5 mL ThinPrep LBC cervical cell material were centrifuged at 5000 g for 30 min in 4 °C, and the cell pellet was resuspended in 400 μL phosphate-buffered saline. DNA was extracted from resuspended cells using MagNA Pure Compact (Roche) and the MagNA Pure Compact Nucleic Acid Isolation Kit (Roche). HPV genotyping was then performed using the Roche Linear Array HPV genotyping test which identifies 37 different high- and low-risk types .Specimen that were positive with the Cobas® HPV test and typed as ‘Other HPV‘but negative with HPVIR were genotyped using the Roche LINEAR ARRAYAll gynaecologic examinations of Swedish samples were performed at the Clinic of Obstetrics and Gynaecology, Uppsala University Hospital. A sample for cytological analysis was also collected at the same visit. The colposcopic evaluation included an identification of squamocolumnar junction and transformation zone (TZ) with application of 5% acetic acid and iodine solution. Directed biopsies were obtained from all the identified abnormal areas and a blind biopsy was taken in women with normal colposcopy. All cytology and histology samples were analysed at the Clinic of Pathology and Cytology, Uppsala University Hospital, Uppsala. The classification was according to SNOMED  and the highest histological grade found in each patient was used for interpretation of the results.https://www.graphpad.com/quickcalcs/kappa1/). The thresholds for intra-laboratory reproducibility and inter-laboratory agreement were set to a lower confidence bound of ≥87% with kappa values > 0.5 as proposed by Meijer and colleagues [Clinical specificity and sensitivity for samples on FTA card analysed with HPVIR was compared with LBC samples analysed with the Cobas® HPV test, using a binomial test for non-inferiority with a power of at least 80%, as estimated in earlier studies , 20. Nonlleagues .® HPV test  compared to using HPVIR . Among women that were positive only with Cobas® HPV test, 42% (11/26) had normal histology and 58% (15/26) had CIN1. The clinical specificity of HPVIR was 96.8% (95% CI = 95.3–97.8) compared to 93.9% (95% CI = 92.0–95.3) for the Cobas® HPV test. There was no statistical evidence that the clinical specificity of HPVIR was inferior to that of the Cobas® HPV test .Two samples, the FTA card and LBC, were obtained from 799 women  with no evidence of ≥ CIN2, and these were used in the calculation of clinical specificity. Of the 799 women, the candidate and reference test agreed on 747 women to be HPV negative and 23 women to be HPV positive, corresponding to an agreement of 96.4% Table . Among t® HPV test. No woman was negative by both tests. The agreement between the methods was 89.6% (60/67). The clinical sensitivity of HPVIR was 95.5% (95% CI = 87.2–99.0) and that of Cobas® HPV was 94.0% (95% CI = 85.2–98.1). In conclusion, there was no statistical evidence that the clinical sensitivity of HPVIR was inferior to that of Cobas® HPV test .To estimate the clinical sensitivity, samples from 67 women ≥30 years  with histologically confirmed ≥ CIN2 were used. Among these, 23 women had CIN2 and 44 had ≥ CIN3 and the median follow-up time was 5.2 (range 0 to 42.2) months. Among the 67 women, both tests agreed on 60 women to be HPV positive and none of the women to be HPV negative Table . Four won = 23) of the LBC samples that were HPV positive with ‘Other HPV’ using Cobas® HPV test but negative with HPVIR were analysed using the Roche LINEAR ARRAY® HPV genotyping test. Of the 23 samples, the linear array was negative for 2 samples, 2 samples had only non-hrHPV types , another 2 samples contained both hrHPV and non-hrHPV  while the remaining 17 samples had one or several of the following hrHPV 31, 33, 35, 39, 45, 52, 58, 59, 66, 68. Among the 23 samples, 6 contained only HPV66 and/or HPV 68.A set . In Uppsala, two research engineers (Operators) performed a blinded intra-laboratory test. The reproducibility  between operators was estimated to 0.974 (95% CI = 0.956–0.993), with an agreement of 98.7%  of discordant samples were HPV positive only with the Cobas® HPV test. In 88.5% (23/26) of these discordant samples, the genotype was indicated as ‘Other HPV‘by the Cobas® HPV test. For two of these samples, the linear array did not indicate the HPV type, possibly due to limited amounts of material. Of the 21 samples that were Cobas® HPV positive but HPVIR negative and for which the linear array resulted in an HPV genotype, 6 of the samples had HPV66 and 68, types not included in the HPVIR test. HPV66 is a genotype that was previously considered high-risk, but recent studies indicate that it has low or no oncogenic properties [® HPV cross-hybridizes. The category ‘Other HPV‘in the Cobas® HPV test has previously been reported to show cross-reactivity with low-risk HPV genotypes. A study of 5022 women comparing two other commercial hrHPV tests with Cobas® HPV showed that about 25% of the samples that were positive with the Cobas® HPV test represented cross-reactivity with low-risk genotypes [® HPV test compared to HPVIR test would increase colposcopy resources and costs for the screening program, as well as worry some women that will be exposed to unnecessary examination.Among women without disease, 3.6% (29/799) showed discordant HPV test results between the HPVIR and the Cobasoperties . The oncIn 88.5% /26 of thenotypes . Thus, t® HPV was similar in the group of cases used to estimate the sensitivity, were none of the samples was HPV negative and 89.6% (60/67) had HPV positive results. Three samples were positive only using the Cobas® HPV test  and 4 samples  were positive only using the HPVIR test. Our results confirms that HPV16 or 18 occurs in more than 50% of women with high-risk lesions [The performance of HPVIR and Cobas lesions .® LBC for the Cobas® HPV test was compared to the indicating FTA Elute™ micro card, the dry sampling medium for the HPVIR test. Since two different samples were collected from each woman, the sampling order may affect the results of the HPV assay. There was no significant difference in HPV positivity between the samples collected first and second for either of the tests. A screening method using a dry storage media as the indicating FTA Elute™ micro card in combination with a highly sensitive HPV test provides an effective method for primary cervical cancer screening. This strategy will present a low cost and high throughput alternative, in addition to being easily adapted for self-sampling. HPV testing using vaginal self-sampling on FTA card has been shown to be as reliable as assisted cervical sampling [In addition to comparing the HPVIR test against a reference test, we used different sampling media. The internationally widely used standardized method ThinPrepsampling , 15. We sampling , 15. Thisampling .The inter- and inter-laboratory analyses was based on 558 samples collected in South Africa selected on the criterion that at least 30% had previously tested HPV positive with the reference test HC2. In both the intra- and inter-laboratory analyses the lower confidence bounds were > 87% with kappa values > 0.5 and, the results complied well with the guidelines. The HPVIR test has been validated yearly using the WHO panel with 100% proficiency. The result from inter-laboratory testing showed that the HPVIR method is robust enough and can be operated reliably at other sites than Uppsala.® HPV test, and thereby fulfil the requirements of the international guidelines for an HPV test to be used for primary cervical cancer screening. The use of the FTA cards and HPVIR has the advantage that it is both easily used for self-sampling and report extended HPV genotyping results.Cervical samples applied on FTA card and analysed with HPVIR test shows similar sensitivity and specificity as LBC samples analysed with the Cobas


Young European Scientist Meeting has its main stage in Porto, Portugal. This year's conference will gather more than 500 students from biomedical areas, who will have the unique chance to present their research findings and to participate in the social programme, scientific sessions, practical workshops, and clinical competition.The 14th edition of the The YES Meeting concept was born in 2006 as medical students felt the need to create a space where they would present their research findings and have the opportunity to contact and chat with scientists and physicians from all over the world. This networking need of Portuguese medical students led to 14th successful editions so far, promoting a conference which grows year after year, being the first Portuguese biomedical congress solely organized by students.Throughout every edition, all YES Meeting Committees worked to shape the conference we see and know today. In 2019, the YES Meeting is happy to receive, in the humble city of Porto, Professor F. Ulrich Hartl, Lasker Award Basic Research winner in 2011, and Professor John van der Oost, Spinoza Prize winner in 2018, as well as experts on robotics, virtual surgery, mental health, melanoma, human genetics, ex-utero devices, among other interesting subjects.Porto Biomedical Journal, which launched the Scientific Competition further away. By visiting the following pages, readers may find all abstracts presented at the 14th YES Meeting, selected out of 262 valid submissions in the fields of Neurosciences, Oncology & Molecular Biology, Public Health & Medical Informatics, Internal Medicine, Immunology & Physiology, and Surgery, as well as articles written by renowned scientists, who attended previous editions of the YES Meeting. On behalf of the Organizing Committee, I would like to thank all the speakers who have embraced this initiative.Since 2017, the YES Meeting has established a prosperous partnership with the Porto Biomedical Journal, to break boundaries one more time.In conclusion, it is my pleasure to invite you all to read this issue's pages, born out a colligation between the YES Meeting and the