SND1 binds SARS-CoV-2 negative-sense RNA and promotes viral RNA synthesis through NSP9.

Regulation of viral RNA biogenesis is fundamental to productive SARS-CoV-2 infection. To characterize host RNA-binding proteins (RBPs) involved in this process, we biochemically identified proteins bound to genomic and subgenomic SARS-CoV-2 RNAs. We find that the host protein SND1 binds the 5' end of negative-sense viral RNA and is required for SARS-CoV-2 RNA synthesis. SND1-depleted cells form smaller replication organelles and display diminished virus growth kinetics. We discover that NSP9, a viral RBP and direct SND1 interaction partner, is covalently linked to the 5' ends of positive- and negative-sense RNAs produced during infection. These linkages occur at replication-transcription initiation sites, consistent with NSP9 priming viral RNA synthesis. Mechanistically, SND1 remodels NSP9 occupancy and alters the covalent linkage of NSP9 to initiating nucleotides in viral RNA. Our findings implicate NSP9 in the initiation of SARS-CoV-2 RNA synthesis and unravel an unsuspected role of a cellular protein in orchestrating viral RNA production.

Fusobacterium and colorectal cancer: causal factor or passenger? Results from a large colorectal cancer screening study.

Colorectal cancer is a leading cause of morbidity and mortality worldwide in both men and women. The gut microbiome is increasingly recognized as having an important role in human health and disease. Fusobacterium has been identified in former studies as a leading gut bacterium associated with colorectal cancer, but it is still not clear if it plays an oncogenic role. In the current study, fecal samples were collected prior to bowel preparation from participants of screening colonoscopy in the German BliTz study. Using 16S rRNA gene analysis, we examined the presence and relative abundance of Fusobacterium in fecal samples from 500 participants, including 46, 113, 110 and 231 individuals with colorectal cancer, advanced adenomas, non-advanced adenomas and without any neoplasms, respectively. We found that the abundance of Fusobacterium in feces was strongly associated with the presence of colorectal cancer (P-value < 0.0001). This was confirmed by PCR at the species level for Fusobacterium nucleatum. However, no association was seen with the presence of advanced adenomas (P-value = 0.80) or non-advanced adenomas (P-value = 0.80), nor were there any associations observed with dietary or lifestyle habits. Although a causal role cannot be ruled out, our observations, based on fecal microbiome, support the hypothesis that Fusobacterium is a passenger that multiplies in the more favorable conditions caused by the malignant tumor rather than a causal factor in colorectal cancer development.

Serology of Streptococcus gallolyticus subspecies gallolyticus and its association with colorectal cancer and precursors.

Streptococcus gallolyticus subspecies gallolyticus (SGG) is potentially associated with colorectal cancer (CRC) and its precursors. A previous case-control study measured antibody responses to SGG pilus proteins Gallo2178 and Gallo2179 and identified significant associations with a small fraction of CRC cases. We aimed at replicating and expanding these findings in an independent study including additional SGG antigens and explored the association with precancerous lesions. We applied multiplex serology to measure antibodies to eleven SGG proteins in serum samples of a screening colonoscopy trial (BliTz study) including participants diagnosed with either non-advanced adenoma (NAA, n = 30), advanced adenoma (AA, n = 100), CRC (n = 50) or controls (n = 228). In addition, we analyzed CRC samples (n = 318) from patients recruited in a clinical setting (DACHSplus study). The association of antibody responses to SGG pilus proteins Gallo2178 and Gallo2179 with CRC was replicated with 4% positive DACHSplus cases compared to 0% positive BliTz controls. Positivity to two or more proteins of a newly defined panel of six SGG markers was significantly associated with CRC in the DACHSplus study (OR: 1.81, 95% CI: 1.07-3.06). Odds for CRC, AA and NAA in the BliTz study were also increased with antibody responses to SGG, and the association was significant for NAA (OR: 2.98, 95% CI: 1.18-7.57). Antibody responses to SGG are associated with CRC and its precursors. The newly identified SGG six-marker panel and associations found with precancerous lesions should be further explored.

Fresh vs Frozen Samples and Ambient Temperature Have Little Effect on Detection of Colorectal Cancer or Adenomas by a Fecal Immunochemical Test in a Colorectal Cancer Screening Cohort in Germany.

BACKGROUND & AIMS: Fecal immunochemical tests (FITs) are used in colorectal cancer (CRC) screening. We compared detection of CRCs and colorectal neoplasms by FITs using fresh samples (collected into buffer-filled tubes) vs frozen samples, and we assessed the effects of seasonal variations in ambient temperature on test performance. METHODS: We performed a prospective study of 3466 individuals (50% male; mean age, 62 years) undergoing screening colonoscopies at 20 gastroenterology practices in southern Germany from November 2008 through September 2014. Frozen stool samples (collected and frozen by patients through February 2012, n = 1644) and fresh stool samples (collected by patients into buffer-filled tubes after February 2012, n = 1822) were obtained; hemoglobin (Hgb) concentrations were measured by using a commercial, quantitative FIT (cutoff value for positive result, 17 µg Hgb/g feces). Colonoscopy results were used as the gold standard, with results categorized as CRC, advanced adenoma, non-advanced adenoma, or no colorectal neoplasm. Differences in detection of colorectal neoplasms with fresh vs frozen samples were compared by using Wilcoxon rank sum test (continuous variables) and Fisher exact test (categorical variables). We also compared test performance when samples were collected during different seasons (based on outdoor temperature less than 8°, 8°-15°, or more than 15°). RESULTS: Of the samples analyzed by FIT, 12.8% of frozen stool samples (95% confidence interval [CI], 11.3%-14.5%) and 8.7% of fresh stool samples (95% CI, 7.5%-10.1%) had positive results (P value for difference < .001). When adjusting the Hgb cutoff value to produce the same percentage of positive results for fresh and frozen samples (10% and 5%), FIT with frozen vs fresh samples detected colorectal neoplasms with similar levels of sensitivity and specificity. For example, at cutoff values that produced 5% positive results for each sample type, FIT detected advanced neoplasms with 27.8% sensitivity when frozen samples were used (95% CI, 21.4%-35.1%) and 25.6% sensitivity when fresh samples were used (95% CI, 19.8%-32.1%). Specificity values were 97.7% when frozen samples were used (95% CI, 96.8%-98.4%) and 97.6% when fresh samples were used (95% CI, 96.7%-98.3%). We did not observe any differences in detection of neoplasms during different seasons that were based on outdoor temperature. CONCLUSIONS: In a prospective study of 3466 individuals who underwent screening colonoscopies and received FITs, we found that use of fresh vs frozen samples slightly affected positivity rates and the proportions of CRCs or adenomas detected at the recommended Hgb cutoff value. However, after we adjusted Hgb cutoff values to produce equal proportions of positive results for fresh vs frozen samples, the performance of the FIT was similar with each sample type. Season of sample collection (based on outdoor temperature) did not affect detection of CRC using either sample type in this study from Middle Europe.

Soybean yield in relation to distance from the Itaipu reservoir.

Crops close to small water bodies may exhibit changes in yield if the water mass causes significant changes in the microclimate of areas near the reservoir shoreline. The scientific literature describes this effect as occurring gradually, with higher intensity in the sites near the shoreline and decreasing intensity with distance from the reservoir. Experiments with two soybean cultivars were conducted during four crop seasons to evaluate soybean yield in relation to distance from the Itaipu reservoir and determine the effect of air temperature and water availability on soybean crop yield. Fifteen experimental sites were distributed in three transects perpendicular to the Itaipu reservoir, covering an area at approximately 10 km from the shoreline. The yield gradient between the site closest to the reservoir and the sites farther away in each transect did not show a consistent trend, but varied as a function of distance, crop season, and cultivar. This finding indicates that the Itaipu reservoir does not affect the yield of soybean plants grown within approximately 10 km from the shoreline. In addition, the variation in yield among the experimental sites was not attributed to thermal conditions because the temperature was similar within transects. However, the crop water availability was responsible for higher differences in yield among the neighboring experimental sites related to water stress caused by spatial variability in rainfall, especially during the soybean reproductive period in January and February.

Systematic review: serum autoantibodies in the early detection of gastric cancer.

Antibodies against tumor-associated antigens have been found in serum of patients with various types of cancers and may serve as biomarkers for early detection of gastric cancer as well. This systematic review aims to give an overview about known autoantibodies and their diagnostic value in gastric cancer. We conducted a systematic literature search in two databases to identify studies which performed serological testing for autoantibodies in gastric cancer patients and controls. Data on study characteristics and results were extracted independently by two reviewers. Overall, 39 articles reporting the detection of 34 different autoantibodies met the inclusion criteria for this review. The most common antibody detection method was enzyme-linked immunosorbent assay and the most frequently assessed autoantibody was anti-p53, which was tested in 13 studies. Most antibodies were assessed in only one study and only few authors have evaluated the diagnostic value of combinations of multiple autoantibodies. For single autoantibodies, specificity was generally very high (median: 99.15%), but sensitivity was mostly rather low (median: 12.35%). For some autoantibody combinations, substantially higher sensitivity at reasonably high levels of specificity could be achieved. Development of extended and optimized multimarker panels of autoantibodies might be a promising approach for gastric cancer early detection.

Occurrence of gastrointestinal parasites in dogs in a rural area of Santa Catarina, Brazil.

We investigated the occurrence of gastrointestinal parasites in dogs in rural areas of the municipality of Painel, Santa Catarina, Brazil. For this, 91 canine feces samples were collected from 82 farms between August, 2017 and January, 2018. These fecal materials were processed using the techniques of spontaneous sedimentation, centrifugation-floatation in zinc sulfate and Ziehl-Neelsen staining. A questionnaire in the form of an interview was administered to the dogs' owner and information about the farm and the main care provided for the dogs was obtained. Among 91 sampled dogs, 27 (29.7%) were positive for at least one parasite species. Ancylostoma was the most prevalent genus (16.5%), followed by Giardia duodenalis (14.3%), Trichuris vulpis (6.6%), Toxocara canis (5.5%), Entamoeba spp. (4.8%), Cryptosporidium spp. (3.3%) and Taeniidae (1.1%). Most dogs do not receive veterinarian care and rarely received antiparasitic treatment. They were free to roam and had free access to animal remains and garbage, which was reflected in the significant associations with the occurrence of parasites that were found. We conclude that rural dogs harbor gastrointestinal parasites, but that their owners are unaware of the risks that these parasites can bring to human health.

Stereotactic radiosurgery and combined immune checkpoint therapy with ipilimumab and nivolumab in patients with melanoma brain metastases: A retrospective monocentric toxicity analysis.

Purpose and objective: Adding stereotactic radiosurgery (SRS) to combined immune checkpoint therapy with ipilimumab and nivolumab (IPI + NIVO) has led to promising results for patients with melanoma brain metastases (MBM). This study retrospectively analyzes the toxicity profile depending on the timing of SRS with regard to IPI + NIVO. Materials and methods: For this study, the clinical database was searched for all patients with MBM who were treated with SRS and IPI + NIVO. The patients were separated into three groups: group A completed IPI + NIVO (usually up to four cycles) >14 days before SRS, in group B IPI + NIVO was initiated>14 days after SRS, and group C received SRS concurrently to IPI + NIVO. Treatment related toxicity was obtained from clinical and neuroradiological records. Analyses were performed using the Fisher-Yates-test. Results: 31 patients were assessed including six (19.4 %), seven (22.6 %) and 18 (58.1 %) patients, in groups A, B and C, respectively. Baseline prognostic markers between groups were balanced. In total, five (16.1 %) patients experienced neurological grade 3 toxicities related to SRS. All of these five patients were in group C, which was near-significantly correlated with a risk for grade 3 toxicities (p = 0.058). Post-hoc analyses showed that a maximum time period of seven days between SRS and IPI + NIVO was significantly correlated with grade 3 toxicity (p = 0.048). Conclusion: Application of SRS to IPI + NIVO within a seven-day span was related to higher toxicity rates in this retrospective analysis. After previous studies focused on immune checkpoint monotherapies with SRS and declared it as safe, this study indicates that concomitant application of IPI + NIVO and SRS might increase side effects. Prospective validation is warranted to corroborate these findings.

Validation of a genotype-based algorithm that identifies individuals with low, intermediate, and high serum CA19-9 levels in cancer-free individuals and in patients with colorectal cancer.

Background: Serum levels of Carbohydrate antigen CA19-9 are determined by the genotype of fucosyltransferases 2 and 3. To validate, possibly modify, and improve a grouping algorithm based on these genotypes. Methods: CA19-9 levels genotypes and of fucosyltransferase 2 and 3 were analyzed in cancer-free and colorectal cancer patients. Patients were assigned to groups with low (group A), intermediate (B), or high (C) CA19-9 biosynthetic activity based on a previously developed grouping algorithm based on genotype of fucosyltransferases 2 and 3. Results: Three hundred thirty-eight patients were included (n=177 cancer-free). Of cancer-free patients 7.9%, 75.7%, and 16.4% were assigned to groups A, B, and C, respectively. In colorectal cancer patients it 7.5%, 77.0%, and 15.5%, respectively. There were significant differences between median CA19-9 levels in the three groups (P<0.001) in both cohorts. The T59G single-nucleotid polymorphism in fucosyltransferase 3 had a significant influence on CA19-9 levels in cancer-free group B patients, which led to establishment of subgroups B1 and B2. However, no difference in CA19-9 levels between these subgroups was found in colorectal cancer patients. A receiver-operating characteristic showed similar areas under the curve for original group B as well as for subgroups B1 and B2. Conclusions: The grouping algorithm based on genotype of fucosyltransferases 2 and 3, which defines groups with distinct CA19-9 serum levels, was validated in cancer-free patients and in colorectal cancer patients. No clinically relevant improvement to the grouping algorithm was identified.

Grazing intensity drives plant diversity but does not affect forage production in a natural grassland dominated by the tussock-forming grass Andropogon lateralis Nees.

Andropogon lateralis is a tall and highly plastic tussock-forming grass native from southern South America. It is a frequent component of Campos and Subtropical highland grasslands that often becomes dominant under lax grazing regimes. The aim of this work was to analyze the response of species diversity and forage production of a natural grassland dominated by A. lateralis to a wide range of grazing intensity. We hypothesized that species diversity and forage production would both peak at the intermediate canopy heights determined by grazing regimes of moderate intensity. A grazing experiment was conducted in a highland grassland with mesothermal humid climate at 922 masl (Atlantic Forest biome, Santa Catarina state, Brazil) that comprised 87 species from 20 families but had 50% of its standing biomass accounted by A. lateralis. Four pre-/post-grazing canopy heights-12/7, 20/12, 28/17, and 36/22 cm (measured on A. lateralis)-were arranged in a complete randomized block design with four replications, and intermittently stocked with beef heifers from October 2015 to October 2017. Andropogon lateralis cover decreased (from 75 to 50%), and species richness increased (15-25 species m-2) as canopy height decreased. Grazing intensity did not affect annual forage production (4.2 Mg DM ha-1). This natural grassland dominated by A. lateralis had a high capacity to adjust to grazing regimes of contrasting intensity, maintaining forage production stable over a wide range of canopy heights. However, to prevent losses in floristic diversity, such grassland should not be grazed at canopy heights higher than 28 cm.

The SARS-CoV-2 RNA-protein interactome in infected human cells.

Characterizing the interactions that SARS-CoV-2 viral RNAs make with host cell proteins during infection can improve our understanding of viral RNA functions and the host innate immune response. Using RNA antisense purification and mass spectrometry, we identified up to 104 human proteins that directly and specifically bind to SARS-CoV-2 RNAs in infected human cells. We integrated the SARS-CoV-2 RNA interactome with changes in proteome abundance induced by viral infection and linked interactome proteins to cellular pathways relevant to SARS-CoV-2 infections. We demonstrated by genetic perturbation that cellular nucleic acid-binding protein (CNBP) and La-related protein 1 (LARP1), two of the most strongly enriched viral RNA binders, restrict SARS-CoV-2 replication in infected cells and provide a global map of their direct RNA contact sites. Pharmacological inhibition of three other RNA interactome members, PPIA, ATP1A1, and the ARP2/3 complex, reduced viral replication in two human cell lines. The identification of host dependency factors and defence strategies as presented in this work will improve the design of targeted therapeutics against SARS-CoV-2.

Variation of diagnostic performance of fecal immunochemical testing for hemoglobin by sex and age: results from a large screening cohort.

OBJECTIVE: Fecal immunochemical tests (FITs) for hemoglobin in stool are increasingly used for colorectal cancer screening. Reported sensitivities and specificities have strongly varied between studies, but it is unclear to what extent such variation reflects differences between tests or between study population characteristics. We aimed to evaluate the key parameters of FIT performance for detecting advanced neoplasia (AN) according to sex and age. METHODS: Sex- and age-specific sensitivity, specificity, positive predictive values (PPVs) and negative predictive values (NPVs) for detecting AN of a quantitative FIT (FOB Gold®) were evaluated among 3211 men and women aged 50-79 years who underwent screening colonoscopy in Germany. RESULTS: At the cutoff recommended by the manufacturer (17 µg hemoglobin/g feces), sensitivity was higher (51.2% versus 34.7%, p=0.004) and specificity was lower (91.0% versus 94.8%, p<0.001) among 65-79 year-old participants compared with 50-64 year-old participants. PPVs and NPVs did not differ significantly between age groups. However, higher NPVs were observed among women compared with men (94.7% versus 92.5%, p=0.015). Specificity was also higher among women compared with men (94.7% versus 92.3%, p=0.007), while there was only a little variation in sensitivity (40.3% versus 41.8%, p=0.789) according to sex. In joint stratification by both factors, sensitivity ranged from 34.1% (95% CI 24.2%-45.2%) in 50-64 year-old women to 51.4% (95% CI 39.3%-63.3%) in 65-79 year-old men (p=0.029). The observed age and sex differences were highly consistent across a wide range of alternative cutoffs from 10 to 50 µg hemoglobin/g feces. CONCLUSION: There are major differences in diagnostic performance parameters according to sex and age, which should receive careful attention in the interpretation and comparison of results of FIT-based colorectal cancer screening studies.

Biomarker discovery study of inflammatory proteins for colorectal cancer early detection demonstrated importance of screening setting validation.

OBJECTIVES: Most studies identifying inflammatory markers for early detection of colorectal cancer (CRC) were conducted using clinically manifest cases. We aimed to identify circulating inflammatory biomarkers for early detection of CRC and validate them in both a clinical setting and a true screening setting. STUDY DESIGN AND SETTING: A total of 92 inflammatory proteins were quantified in baseline plasma samples from individuals clinically diagnosed with CRC and neoplasm-free controls matched on age and sex (training set). A multimarker panel was selected and evaluated in samples from another clinical setting (validation set C) and a screening setting (validation set S). RESULTS: In the training set (N = 330), a five-biomarker signature was selected that provided an area under curve (AUC) of 0.85 and 60.9% sensitivity to detect CRC at 90% specificity. When this algorithm was applied to validation set C (N = 318), the AUC (0.80) and sensitivity (49.5%) at 90% specificity for CRC diagnosis were only slightly lower than those in the training set. By contrast, the diagnostic performance of the algorithm in validation set S (N = 126) from a true screening setting was much poorer, with an AUC of 0.59 and a sensitivity of 28.6% at 90% specificity. CONCLUSIONS: An inflammation-related protein panel with apparently good diagnostic properties for CRC detection was identified and confirmed in an independent clinical validation set. However, the biomarker combination performed substantially worse in a validation sample from a true screening setting. Our results underline the importance of validation in screening settings subsequently to novel signature discovery for cancer early detection.

Occurrence of gastrointestinal parasites in dogs in a rural area of Santa Catarina, Brazil / Ocorrência de parasitas gastrintestinais em cães de área rural de Santa Catarina, Brasil

Abstract We investigated the occurrence of gastrointestinal parasites in dogs in rural areas of the municipality of Painel, Santa Catarina, Brazil. For this, 91 canine feces samples were collected from 82 farms between August, 2017 and January, 2018. These fecal materials were processed using the techniques of spontaneous sedimentation, centrifugation-floatation in zinc sulfate and Ziehl-Neelsen staining. A questionnaire in the form of an interview was administered to the dogs' owner and information about the farm and the main care provided for the dogs was obtained. Among 91 sampled dogs, 27 (29.7%) were positive for at least one parasite species. Ancylostoma was the most prevalent genus (16.5%), followed by Giardia duodenalis (14.3%), Trichuris vulpis (6.6%), Toxocara canis (5.5%), Entamoeba spp. (4.8%), Cryptosporidium spp. (3.3%) and Taeniidae (1.1%). Most dogs do not receive veterinarian care and rarely received antiparasitic treatment. They were free to roam and had free access to animal remains and garbage, which was reflected in the significant associations with the occurrence of parasites that were found. We conclude that rural dogs harbor gastrointestinal parasites, but that their owners are unaware of the risks that these parasites can bring to human health.

Resumo Foi investigada a ocorrência de parasitos gastrintestinais com potencial zoonótico, em cães de área rural do munícipio de Painel, Santa Catarina, Brasil. Para isso, foram colhidas amostras de fezes de 91 cães de 82 propriedades rurais, no período de agosto de 2017 a janeiro de 2018. Este material fecal foi processado pelas técnicas de sedimentação espontânea, centrífugo-flutuação em sulfato de zinco e coloração pelo Ziehl-Neelsen. Um questionário em forma de entrevista foi aplicado aos tutores dos cães e foram obtidas informações sobre a propriedade rural e principais cuidados com os cães. Entre as 91 amostras de fezes, em 27 (29,7%) foi detectada a presença de parasitos gastrintestinais, sendo Ancylostoma o gênero mais prevalente (15 / 16,5%), seguido por Giardia duodenalis (13 / 14,3%), Trichuris vulpis (6 / 6,6%), Toxocara canis (5 / 5,5%), Entamoeba spp. (4 / 4,8%), Cryptosporidium spp. (3 / 3,3%) e Taeniidae (1 / 1,1%). A maioria dos cães não recebia cuidados veterinários ou antiparasitários. Concluiu-se que os cães de área rural albergam parasitos gastrintestinais, porém seus tutores desconhecem os riscos que esses parasitos podem trazer para a saúde humana.

First record of in vitro formation of ectomycorrhizae in Psidium cattleianum Sabine, a native Myrtaceae of the Brazilian Atlantic Forest.

Like many other species of trees native to the Brazilian Mata Atlântica (Atlantic Forest), the Myrtaceae, such as the Red Araza (Psidium cattleianum Sabine), are widely cited as arbuscular mycorrhizal formers. Nevertheless, recent studies show evidence that Myrtaceae from different tropical, subtropical and neotropical ecosystems can also prompt the formation of ectomycorrhizae, indicating that this species' ectomycorrhizal status should be further explored. Because of this, this research effort studied the in vitro interaction between the Red Araza and two ectomycorrhizal fungi isolates, belonging to the Pisolithus microcarpus (D17) and Scleroderma citrinum (UFSC-Sc133) species. An analysis was performed to determine the formation of ectomycorrhizal structures, or lack thereof, and the developmental differences between the in vitro mycorrhized and non-mycorrhized plants. The analysis proved that indeed an ectomycorrhizal association was developed between the Red Araza, and the D17 and UFSC-Sc133 isolates, a fact never before registered in the existing literature. After an in vitro period of 110 days, it was confirmed that the D17 and UFSC-Sc133 isolates formed mycorrhizal colonization of 91.6% and 15.7%, respectively. Furthermore, both isolates also promoted root thickening, and the formation of a fungal mantle and a Hartig net. However, when compared to the Control plants, the fungal isolates did not contribute to an increase in the development of the subject plants, possibly due to the specific experimental conditions used, such as a high humidity environment and high availability of nutrients in the symbiotic substrate.

Selecting a Cut-off for Colorectal Cancer Screening With a Fecal Immunochemical Test.

OBJECTIVES: Fecal immunochemical tests (FITs) for hemoglobin (Hb) are increasingly used for colorectal cancer (CRC) screening. However, cut-offs for defining test positivity are varying widely. We aimed to evaluate the impact of cut-off selection on key indicators of diagnostic performance in a true screening setting. METHODS: We evaluated diagnostic performance of FOB Gold, a widely used quantitative FIT, for detecting advanced neoplasms (AN) across a wide range of possible cut-offs among 1822 participants of screening colonoscopy aged 50-79 years in Germany. RESULTS: The positive predictive value (PPV) for detecting AN showed a very steep increase with increasing cut-off up to 35.2% (95% CI 29.9-40.9%) at a cut-off of 9 µg Hb/g feces at which sensitivity and specificity were 48.8% (95% CI 42.1-55.6%) and 88.5% (95% CI 86.8-89.9%), respectively. A further moderate increase of PPV up to 56.9% (95% CI 47.8-65.5%), along with a major decrease in sensitivity was observed when gradually increasing the cut-off to 25 µg Hb/g feces at which sensitivity and specificity were 31.9% (95% CI 25.9-38.5%) and 96.9% (95% CI 95.9-97.6%), respectively. Further increases of the cut-off hardly affected PPV and specificity, but went along with further relevant decline in sensitivity. CONCLUSIONS: Our study illustrates delineation of a range of meaningful cut-offs (here: 9-25 µg Hb/g feces) according to expected diagnostic yield in a true screening setting. Selecting a cut-off within or beyond this range should consider characteristics of the specific target population, such as AN prevalence or available colonoscopy capacity.

Development and validation of a panel of five proteins as blood biomarkers for early detection of colorectal cancer.

OBJECTIVE: Reliable noninvasive biomarkers for early detection of colorectal cancer (CRC) are highly desirable for efficient population-based screening with high adherence rates. We aimed to discover and validate blood-based protein markers for the early detection of CRC. PATIENTS AND METHODS: A two-stage design with a discovery and a validation set was used. In the discovery phase, plasma levels of 92 protein markers and serum levels of TP53 autoantibody were measured in 226 clinically recruited CRC patients and 118 controls who were free of colorectal neoplasms at screening colonoscopy. An algorithm predicting the presence of CRC was derived by Lasso regression and validated in a validation set consisting of all available 41 patients with CRC and a representative sample of 106 participants with advanced adenomas and 107 controls free of neoplasm from a large screening colonoscopy cohort (N=6018). Receiver operating characteristic (ROC) analyses were conducted to evaluate the diagnostic performance of individual biomarkers and biomarker combinations. RESULTS: An algorithm based on growth differentiation factor 15 (GDF-15), amphiregulin (AREG), Fas antigen ligand (FasL), Fms-related tyrosine kinase 3 ligand (Flt3L) and TP53 autoantibody was constructed. In the validation set, the areas under the curves of this five-marker algorithm were 0.82 (95% CI, 0.74-0.90) for detecting CRC and 0.60 (95% CI, 0.52-0.69) for detecting advanced adenomas. At cutoffs yielding 90% specificity, the sensitivities (95% CI) for detecting CRC and advanced adenomas were 56.4% (38.4%-71.8%) and 22.0% (13.4%-35.4%), respectively. The five-marker panel showed similar diagnostic efficacy for the detection of early- and late-stage CRC. CONCLUSION: The identified most promising biomarkers could contribute to the development of powerful blood-based tests for CRC screening in the future.

Blood-Based Protein Signatures for Early Detection of Colorectal Cancer: A Systematic Review.

OBJECTIVES: Blood-based proteins might be an attractive option for early detection of colorectal cancer (CRC), but individually they are unlikely to achieve the diagnostic performance required for population based screening. We aimed at summarizing current evidence of diagnostic performance of signatures based on multiple proteins for early detection of CRC. METHODS: A systematic literature review adhering to the PRISMA (preferred reporting items for systematic reviews and meta-analysis) guidelines was performed. PubMed and Web of Science databases were searched for potentially relevant studies published until 28th August, 2017. Relevant studies were identified by predefined eligibility criteria. Estimates of indicators of diagnostic performance such as sensitivity, specificity, and the area under the curve (AUC), along with information on validation and other key methodological procedures were extracted. Study quality was assessed by a QUADAS-2 (Quality Assessment of Diagnostic Accuracy Studies 2) instrument tool. RESULTS: Thirty six eligible studies with numbers of CRC cases ranging from 23 to 512 and the number of proteins included in signatures ranged from 3 to 13 were identified. Reported Youden's Index and AUC ranged from 0.19 to 0.95 and from 0.62 to 0.996, respectively. However most studies, especially those reporting better diagnostic performance, were conducted in clinical rather than screening setting and many studies lacked any internal or external validation of identified algorithm. CONCLUSIONS: Blood-based tests using signatures of multiple proteins may be a promising approach for non-invasive CRC screening. However, promising signatures identified in clinical settings still require rigorous evaluation in large studies conducted in true screening setting.

Influence of preoperative pain in the success rate of indirect pulp capping: a retrospective study.

BACKGROUND: The association of preoperative pain and indirect pulp treatment in permanent teeth is still unknown. This study aimed to evaluate the influence of preoperative pain in the success rate of indirect pulp capping. METHODS: The sample of this retrospective study was constituted for 133 teeth treated in a Private clinic in Joinvile, Brazil, by one experienced dentist between 2007 and 2012. Data related to clinical and demographic variables that could influence in the success rate of indirect pulp capping were also collected, such patient age and sex, type of teeth and arch. Chi­square test was used to evaluate the association between preoperative pain with postoperative one and success of the treatment. Logistic regression was performed to identify significant clinical and demographical factors associated with the success of the indirect pulp capping. RESULTS: The overall success rate was 100% in the absence of preoperative pain. However, when the preoperative pain was present, the failure rate was 18.3%. Statistical analysis showed significant association between preoperative pain and postoperative one and failure rate of the indirect pulp capping. The other independent variables were not associated with the success rate of the treatment. CONCLUSIONS: The presence of preoperative pain is associated with the decrease of the success rate of indirect pulp capping.