Multi-Omic Analysis of Esophageal Adenocarcinoma Uncovers Candidate Therapeutic Targets and Cancer-Selective Posttranscriptional Regulation.

Efforts to address the poor prognosis associated with esophageal adenocarcinoma (EAC) have been hampered by a lack of biomarkers to identify early disease and therapeutic targets. Despite extensive efforts to understand the somatic mutations associated with EAC over the past decade, a gap remains in understanding how the atlas of genomic aberrations in this cancer impacts the proteome and which somatic variants are of importance for the disease phenotype. We performed a quantitative proteomic analysis of 23 EACs and matched adjacent normal esophageal and gastric tissues. We explored the correlation of transcript and protein abundance using tissue-matched RNA-seq and proteomic data from seven patients and further integrated these data with a cohort of EAC RNA-seq data (n = 264 patients), EAC whole-genome sequencing (n = 454 patients), and external published datasets. We quantified protein expression from 5879 genes in EAC and patient-matched normal tissues. Several biomarker candidates with EAC-selective expression were identified, including the transmembrane protein GPA33. We further verified the EAC-enriched expression of GPA33 in an external cohort of 115 patients and confirm this as an attractive diagnostic and therapeutic target. To further extend the insights gained from our proteomic data, an integrated analysis of protein and RNA expression in EAC and normal tissues revealed several genes with poorly correlated protein and RNA abundance, suggesting posttranscriptional regulation of protein expression. These outlier genes, including SLC25A30, TAOK2, and AGMAT, only rarely demonstrated somatic mutation, suggesting post-transcriptional drivers for this EAC-specific phenotype. AGMAT was demonstrated to be overexpressed at the protein level in EAC compared to adjacent normal tissues with an EAC-selective, post-transcriptional mechanism of regulation of protein abundance proposed. Integrated analysis of proteome, transcriptome, and genome in EAC has revealed several genes with tumor-selective, posttranscriptional regulation of protein expression, which may be an exploitable vulnerability.

IFITM proteins: Understanding their diverse roles in viral infection, cancer, and immunity.

Interferon-induced transmembrane proteins (IFITMs) are broad spectrum antiviral factors that inhibit the entry of a wide range of clinically important pathogens including influenza A virus, HIV-1, and Dengue virus. IFITMs are thought to act primarily by antagonizing virus-cell membrane fusion in this regard. However, recent work on these proteins has uncovered novel post-entry viral restriction mechanisms. IFITMs are also increasingly thought to have a role regulating immune responses, including innate antiviral and inflammatory responses as well as adaptive T-cell and B-cell responses. Further, IFITMs may have pathological activities in cancer, wherein IFITM expression can be a marker of therapeutically resistant and aggressive disease courses. In this review, we summarize the respective literatures concerning these apparently diverse functions with a view to identifying common themes and potentially yielding a more unified understanding of IFITM biology.

n-Type Fullerene-Carbon Dots: Synthesis and Electrochemical and Photophysical Properties.

The covalent incorporation of C60 and C70 derivatives of the well-known n-type organic semiconductor PCBM ([6,6]-phenyl-C61-butyric acid methyl ester) onto carbon dots (CD) is described. Morphological and structural characterization reveal combined features of both pristine starting materials (CD and PCBM). Electrochemical investigations evidenced the existence of additional reduction processes to that of CD or PCBM precursors, showing rich electron-acceptor capabilities, with multistep processes in an affordable and narrow electrochemical window (ca. 1.5 V). Electronic communication in the obtained nanoconjugated species were derived from steady-state absorption and emission spectroscopies, which showed bathochromically shifted absorptions and emissions well entering the red region. Finally, the lower fluorescence quantum yield of CD-PCBM nanoconjugates, compared with CD, and the fast decay of the observed emission of CD, support the existence of an electronic communication between both CD and PCBM units in the excited state.

The Role of IFITM Proteins in Tick-Borne Encephalitis Virus Infection.

Tick-borne encephalitis virus (TBEV), of the genus Flavivirus, is a causative agent of severe encephalitis in regions of endemicity of northern Asia and central and northern Europe. Interferon-induced transmembrane proteins (IFITMs) are restriction factors that inhibit the replication cycles of numerous viruses, including flaviviruses such as West Nile virus, dengue virus, and Zika virus. Here, we demonstrate the role of IFITM1, IFITM2, and IFITM3 in the inhibition of TBEV infection and in protection against virus-induced cell death. We show that the most significant role is that of IFITM3, including the dissection of its functional motifs by mutagenesis. Furthermore, through the use of CRISPR-Cas9-generated IFITM1/3-knockout monoclonal cell lines, we confirm the role and additive action of endogenous IFITMs in TBEV suppression. However, the results of coculture assays suggest that TBEV might partially escape interferon- and IFITM-mediated suppression during high-density coculture infection when the virus enters naive cells directly from infected donor cells. Thus, cell-to-cell spread may constitute a strategy for virus escape from innate host defenses. IMPORTANCE TBEV infection may result in encephalitis, chronic illness, or death. TBEV is endemic in northern Asia and Europe; however, due to climate change, new centers of endemicity have arisen. Although effective TBEV vaccines have been approved, vaccination coverage is low, and due to the lack of specific therapeutics, infected individuals depend on their immune responses to control the infection. IFITM proteins are components of the innate antiviral defenses that suppress cell entry of many viral pathogens. However, no studies on the role of IFITM proteins in TBEV infection have been published thus far. Understanding antiviral innate immune responses is crucial for the future development of antiviral strategies. Here, we show the important role of IFITM proteins in the inhibition of TBEV infection and virus-mediated cell death. However, our data suggest that TBEV cell-to-cell spread may be less prone to both interferon- and IFITM-mediated suppression, potentially facilitating escape from IFITM-mediated immunity.

Factor XI in Carriers of Antiphospholipid Antibodies: Elevated Levels Associated with Symptomatic Thrombotic Cases, While Low Levels Linked to Asymptomatic Cases.

Antiphospholipid syndrome (APS) is a thromboinflammatory disorder caused by circulating antiphospholipid autoantibodies (aPL) and characterized by an increased risk of thrombotic events. The pathogenic mechanisms of these antibodies are complex and not fully understood, but disturbances in coagulation and fibrinolysis have been proposed to contribute to the thrombophilic state. This study aims to evaluate the role of an emerging hemostatic molecule, FXI, in the thrombotic risk of patients with aPL. Cross-sectional and observational study of 194 consecutive and unrelated cases with aPL recruited in a single center: 82 asymptomatic (AaPL) and 112 with primary antiphospholipid syndrome (APS). Clinical and epidemiological variables were collected. The profile of aPL was determined. Plasma FXI was evaluated by Western blotting and two coagulation assays (FXI:C). In cases with low FXI, molecular analysis of the F11 gene was performed. FXI:C levels were significantly higher in patients with APS than in patients with AaPL (122.8 ± 33.4 vs. 104.5 ± 27.5; p < 0.001). Multivariate analysis showed a significant association between symptomatic patients with aPL (APS) and high FXI (>150%) (OR = 11.57; 95% CI: 1.47-90.96; p = 0.020). In contrast, low FXI (<70%), mostly caused by inhibitors, was less frequent in the group of patients with APS compared to AaPL (OR = 0.17; 95%CI: 0.36-0.86; p = 0.032). This study suggests that FXI levels may play a causal role in the prothrombotic state induced by aPLs and holds the promise of complementary treatments in APS patients by targeting FXI.

Phylogenomic analyses of mud dragons (Kinorhyncha).

Mud dragons (Kinorhyncha) are microscopic invertebrates, inhabiting marine sediments across the globe from intertidal to hadal depths. They are segmented, moulting animals like arthropods, but grouping with the unsegmented priapulans and loriciferans within Ecdysozoa. There are more than 300 species of kinorhynchs described within 31 genera and 11 families, however, their evolutionary relationships have so far only been investigated using morphology and a few molecular markers. Here we aim to resolve the relationships and classification of major clades within Kinorhyncha using transcriptomic data. In addition, we wish to revisit the position of three indistinctly segmented, aberrant genera in order to reconstruct the evolution of distinct segmentation within the group. We conducted a phylogenomic analysis of Kinorhyncha including 21 kinorhynch transcriptomes (of which 18 are new) representing 15 genera, and seven outgroups including priapulan, loriciferan, nematode and nematomorph transcriptomes. Results show a congruent and robust tree that supports the division of Kinorhyncha into two major clades: Cyclorhagida and Allomalorhagida. Cyclorhagida is composed of three subclades: Xenosomata, Kentrorhagata comb. nov. (including the aberrant Zelinkaderes) and Echinorhagata. Allomalorhagida is composed of two subclades: Pycnophyidae and Anomoirhaga nom. nov. Anomoirhaga nom. nov. accommodates the aberrant genera Cateria (previously nested within Cyclorhagida) and Franciscideres together with five additional genera. The distant and derived positions of the aberrant Zelinkaderes, Cateria and Franciscideres species suggest that their less distinct trunk segmentation evolved convergently, and that segmentation evolved among kinorhynch stem groups.

Revisiting kinorhynch segmentation: variation of segmental patterns in the nervous system of three aberrant species.

BACKGROUND: Kinorhynch segmentation differs from the patterns found in Chordata, Arthropoda and Annelida which have coeloms and circulatory systems. Due to these differences and their obsolete status as 'Aschelminthes', the microscopic kinorhynchs are often not acknowledged as segmented bilaterians. Yet, morphological studies have shown a conserved segmental arrangement of ectodermal and mesodermal organ systems with spatial correspondence along the anterior-posterior axis. However, a few aberrant kinorhynch lineages present a worm-like body plan with thin cuticle and less distinct segmentation, and thus their study may aid to shed new light on the evolution of segmental patterns within Kinorhyncha. RESULTS: Here we found the nervous system in the aberrant Cateria styx and Franciscideres kalenesos to be clearly segmental, and similar to those of non-aberrant kinorhynchs; hereby not mirroring their otherwise aberrant and posteriorly shifted myoanatomy. In Zelinkaderes yong, however, the segmental arrangement of the nervous system is also shifted posteriorly and misaligned with respect to the cuticular segmentation. CONCLUSIONS: The morphological disparity together with the distant phylogenetic positions of F. kalenesos, C. styx and Z. yong support a convergent origin of aberrant appearances and segmental mismatches within Kinorhyncha.

Insights into the Morphology of Haplozoan Parasites (Dinoflagellata) using Confocal Laser Scanning Microscopy.

We describe new insights into the morphology and life history of the bizarre parasite Haplozoon axiothellae (Dinoflagellata) using light microscopy (LM), scanning electron microscopy (SEM), and confocal laser scanning microscopy (CLSM). Trophonts were isolated from the intestines of host maldanid polychaetes, Axiothella rubrocincta, collected from San Juan Island, Washington, USA. LM and SEM confirmed features previously observed, such as amphiesmal projections, mature and immature junctions between the nucleated compartments of the vermiform syncytium and visible polygonal alveoli. CLSM of adult trophonts fluorescently stained for DNA, tubulin, centrin, and plasma membrane demonstrated several new ultrastructural traits: (1) an extensive basket of parallel microtubules within the trophomere used for host attachment, (2) two physically separated MTOCs (i.e. putative pairs of basal bodies) beneath pores on the ventral side of each compartment, (3) robust mitotic and/or meiotic spindles associated with one to four nuclei in each compartment, (4) spindles with polar bodies that are disconnected from the MTOCs, (5) a centrin-stained fibril within the trophomeres that potentially functions to retract the motile stylet, and (6) cytokinesis in the posterior-most compartments. This study renames haplozoan compartments using the suffix "-mere" rather than "-cyte" (i.e. trophomere, gonomere, sporomere) to reflect their status within a single syncytium.

Application of sunflower pectin gels with low glycemic index in the coating of fresh strawberries stored in modified atmospheres.

BACKGROUND: This study reports the use of low glycemic sunflower pectin gel, elaborated with calcium and without or with sweeteners (sucrose, stevia and saccharin) as an edible coating and its possible combination with two modified atmosphere packaging (MAP), in order to extend shelf life, maintaining the quality, of strawberries during the storage at 4 °C. RESULTS: This pectin coating, formed with only calcium and/or stevia or saccharin, extended the shelf life of strawberries with respect to uncoating fruits, up to 12 days, keeping the microbial load constant, the firmness and less weight loss. With the same edible coatings, the shelf life of strawberries was extended up to 23 days when they were combined with MAP [10% carbon dioxide (CO2 ), 85% nitrogen (N2 ) and 5% oxygen (O2 )], maintaining the quality of strawberries, while the other MAP, with a higher CO2 concentration (20% CO2 , 75% N2 and 5% O2 ), had no effect. CONCLUSIONS: These results highlight the suitability of the combination of edible pectin coating combined with MAP to obtain an important shelf life extension, maintaining the good quality of the fruit. © 2021 Society of Chemical Industry.

Chiral Molecular Carbon Nanostructures.

Chirality is a fascinating property present in naturally occurring and artificial molecules and materials, observable as chiroptical behavior. The emerging area of carbon nanostructures has undergone tremendous development, with a wide variety of carbon nanoforms reported over the last two decades. However, despite interest in merging chirality and nanocarbons, this has been successfully achieved only in empty fullerenes, whereas in other kinds of fullerenes or carbon nanostructures such as carbon nanotubes, graphene, and graphene quantum dots (GQDs), to name the most popular systems, it is almost unknown. Therefore, controlling chirality in carbon nanostructures currently represents a major challenge for the chemical community. In this Account, we show our progress in the synthesis of chiral molecular carbon nanostructures, namely, metallofullerenes, endohedral fullerenes, GQDs, and curved molecular nanographenes, by using asymmetric catalysis and both top-down and bottom-up chemical approaches. Furthermore, we bring in a new family of lesser-known molecular chiral bilayer nanographenes, where chirality is introduced from the starting helicene moiety and a single enantiomer of the nanographene is synthesized. Some important landmarks in the development of chiral molecular carbon nanostructures shown in this Account are the application of synthesis-tailored, enantiomerically pure metallofullerenes as catalysts for hydrogen transfer reactions and the use of endohedral fullerenes to determine the effect of the incarcerated molecule in the carbon cage on the cis-trans stereoisomerization of optically active pendent moieties. Furthermore, the first top-down synthesis of chiral GQDs by functionalization with chiral alcohols is also presented. An emerging alternative to GQDs, when the desire for purity and atomistic control outweighs the cost of multistep synthesis, is the bottom-up approach, in which molecular nanographenes are formed in precise sizes and shapes and enantiomeric control is feasible. In this regard, a singular and amazing example is given by our synthesis of a single enantiomer of the first chiral bilayer nanographene, which formally represents a new family of molecular nanographenes with chirality controlled and maintained throughout their syntheses. The aforementioned synthetic chiral nanostructures represent groundbreaking nanocarbon systems where chirality is a further dimension of structural control, paving the way to a new scenario for carbon nanoforms in which chirality selection determines the properties of these novel carbon-based materials. Fine-tuning of such properties is envisioned to impact biomedical and materials science applications.

Neuroanatomy of mud dragons: a comprehensive view of the nervous system in Echinoderes (Kinorhyncha) by confocal laser scanning microscopy.

BACKGROUND: The Scalidophora (Kinorhyncha, Loricifera and Priapulida) have an important phylogenetic position as early branching ecdysozoans, yet the architecture of their nervous organ systems is notably underinvestigated. Without such information, and in the absence of a stable phylogenetic context, we are inhibited from producing adequate hypotheses about the evolution and diversification of ecdysozoan nervous systems. Here, we utilize confocal laser scanning microscopy to characterize serotonergic, tubulinergic and FMRFamidergic immunoreactivity patterns in a comparative neuroanatomical study with three species of Echinoderes, the most speciose, abundant and diverse genus within Kinorhyncha. RESULTS: Neuroanatomy in Echinoderes as revealed by acetylated α-tubulin immunoreactivity includes a circumpharyngeal brain and ten neurite bundles in the head region that converge into five longitudinal nerves within the trunk. The ventral nerve cord is ganglionated, emerging from the brain with two connectives that converge in trunk segments 2-3, and diverge again within segment 8. The longitudinal nerves and ventral nerve cord are connected by two transverse neurites in segments 2-9. Differences among species correlate with the number, position and innervation of cuticular structures along the body. Patterns of serotoninergic and FMRFamidergic immunoreactivity correlate with the position of the brain neuropil and the ventral nerve cord. Distinct serotonergic and FMRFamidergic somata are associated with the brain neuropil and specific trunk segments along the ventral nerve cord. CONCLUSIONS: Neural architecture is highly conserved across all three species, suggesting that our results reveal a pattern that is common to more than 40% of the species within Kinorhyncha. The nervous system of Echinoderes is segmented along most of the trunk; however, posterior trunk segments exhibit modifications that are likely associated with sensorial, motor or reproductive functions. Although all kinorhynchs show some evidence of an externally segmented trunk, it is unclear whether external segmentation matches internal segmentation of nervous and muscular organ systems across Kinorhyncha, as we observed in Echinoderes. The neuroanatomical data provided in this study not only expand the limited knowledge on kinorhynch nervous systems but also establish a comparative morphological framework within Scalidophora that will support broader inferences about the evolution of neural architecture among the deepest branching lineages of the Ecdysozoa.

Validation of a universal set of primers to study animal-associated microeukaryotic communities.

The application of metabarcoding to study animal-associated microeukaryotes has been restricted because the universal barcode used to study microeukaryotic ecology and distribution in the environment, the Small Subunit of the Ribosomal RNA gene (18S rRNA), is also present in the host. As a result, when host-associated microbial eukaryotes are analysed by metabarcoding, the reads tend to be dominated by host sequences. We have done an in silico validation against the SILVA 18S rRNA database of a non-metazoan primer set (primers that are biased against the metazoan 18S rRNA) that recovers only 2.6% of all the metazoan sequences, while recovering most of the other eukaryotes (80.4%). Among metazoans, the non-metazoan primers are predicted to amplify 74% of Porifera sequences, 4% of Ctenophora, and 15% of Cnidaria, while amplifying almost no sequences within Bilateria. In vivo, these non-metazoan primers reduce significantly the animal signal from coral and human samples, and when compared against universal primers provide at worst a 2-fold decrease in the number of metazoan reads and at best a 2800-fold decrease. This easy, inexpensive, and near-universal method for the study of animal-associated microeukaryotes diversity will contribute to a better understanding of the microbiome.

Anion-π Catalysis on Carbon Nanotubes.

Induced π acidity from polarizability is emerging as the most effective way to stabilize anionic transition states on aromatic π surfaces, that is, anion-π catalysis. To access extreme polarizability, we propose a shift from homogeneous toward heterogeneous anion-π catalysis on higher carbon allotropes. According to benchmark enolate addition chemistry, multi-walled carbon nanotubes equipped with tertiary amine bases outperform single-walled carbon nanotubes. This is consistent with the polarizability of the former not only along but also between the tubes. Inactivation by π-basic aromatics and saturation with increasing catalyst concentration support that catalysis occurs on the π surface of the tubes. Increasing rate and selectivity of existing anion-π catalysts on the surface of unmodified nanotubes is consistent with transition-state stabilization by electron sharing into the tubes, i.e., induced anion-π interactions. On pristine tubes, anion-π catalysis is realized by non-covalent interfacing with π-basic pyrenes.

Circulating microRNAs as biomarkers of disease and typification of the atherothrombotic status in antiphospholipid syndrome.

We aimed to identify the plasma miRNA profile of antiphospholipid syndrome (APS) patients and to investigate the potential role of specific circulating miRNAs as non-invasive disease biomarkers. Ninety APS patients and 42 healthy donors were recruited. Profiling of miRNAs by PCR-array in plasma of APS patients identified a set of miRNAs differentially expressed and collectively involved in clinical features. Logistic regression and ROC analysis identified a signature of 10 miRNA ratios as biomarkers of disease. In addition, miRNA signature was related to fetal loss, atherosclerosis, and type of thrombosis, and correlated with parameters linked to inflammation, thrombosis, and autoimmunity. Hard clustering analysis differentiated 3 clusters representing different thrombotic risk profile groups. Significant differences between groups for several miRNA ratios were found. Moreover, miRNA signature remained stable over time, demonstrated by their analysis three months after the first sample collection. Parallel analysis in two additional cohorts of patients, including thrombosis without autoimmune disease, and systemic lupus erythematosus without antiphospholipid antibodies, each displayed specific miRNA profiles that were distinct from those of APS patients. In vitro, antiphospholipid antibodies of IgG isotype promoted deregulation in selected miRNAs and their potential atherothrombotic protein targets in monocytes and endothelial cells. Taken together, differentially expressed circulating miRNAs in APS patients, modulated at least partially by antiphospholipid antibodies of IgG isotype, might have the potential to serve as novel biomarkers of disease features and to typify patients' atherothrombotic status, thus constituting a useful tool in the management of the disease.

Coronary vascular disease event risk and metabolic syndrome prevalence in patients enrolled in an assertive treatment community program.

OBJECTIVE: To examine the risk of coronary vascular disease event (CVDE) and the prevalence metabolic syndrome (MS) and its cardiovascular risk factors (CVRF) in patients with severe mental illnesses enrolled in an assertive treatment community program (ATC) in Spain. METHOD: We carried out a cross-sectional descriptive study with all of the patients included in an ATC program in 2016 in a health area with 547,328 inhabitants in Galicia, Spain. We identified the CVRF in all the individuals, and calculated MS and 10-year CVDE. We also compared the prevalence of all traits in our cohort and the general population. RESULTS: The 10-year median of coronary vascular disease event (CVDE10) was 8.4%. The percentage of individuals with high CVDE10 (>5%) was 41.2% The CVDE10 median was higher in men than women (10.5% vs 5.1%, p<0.001). MS was detected in 50% of patients without differences between men and women (51.2% vs 48.2%). A prevalence of 68% was found for smoking, 55% for dyslipidemia, 47% for obesity, 29% for impaired glucose metabolism, and 38% for hypertension. Women showed a higher prevalence of obesity measured by elevated waist circumference (88.9% vs 55.6%, p=0.003). Men showed a higher prevalence of arterial hypertension (46.6% vs 22.2%, p=0.0001). CONCLUSIONS: The SMD Patients enrolled in ATC programs had a 1.5-times higher prevalence of MS and 8 times higher CVDE10 than those reported in the general population. Individual CVRF were also higher in the SMD patients. Prevention, early detection, and comprehensive treatment are important issues for patients with severe mental illnesses.

Fourteen-day high-dose esomeprazole, amoxicillin and metronidazole as third-line treatment for Helicobacter pylori infection.

INTRODUCTION: The efficacy of currently recommended third-line therapies for Helicobacter pylori is suboptimal, even that of culture-guided treatments. Resistance to multiple antibiotics is the major factor related to treatment failure. The aim of this study was to evaluate the effectiveness and safety of a 14-day therapy using high-dose of amoxicillin, metronidazole and esomeprazole. MATERIAL AND METHODS: Multicenter open-label study as a register in routine clinical practice in patients with two previous failures of eradication therapy. A triple therapy with esomeprazole 40 mg b.d., amoxicillin 1 g t.d.s and metronidazole 500 mg t.d.s for 2 weeks was administered as a third-line therapy after a first treatment including clarithromycin and a second treatment including a quinolone. Helicobacter pylori status was determined by either histology or 13 C-UBT both before and after treatment. RESULTS: A total of 68 patients were included in this study. An interim analysis showed that only three out of eight patients who had received metronidazole in previous eradication regimens were cured (37%, 95% CI 8-75); as a result, after this interim analysis only metronidazole-naïve patients were included. The ITT eradication rate in metronidazole-naive patients was 64% (95% CI 51-76). Adverse events occurred in 58% of patients, all of them mild-to-moderate. Two patients (3%) did not complete >90% of the treatment because of side effects. No severe adverse events occurred. CONCLUSION: Cure rates of this 14-day schedule using high-dose esomeprazole, amoxicillin and metronidazole as a third-line eradication regimen were suboptimal, especially in patients who had received metronidazole in previous failed eradication regimens.

Management of Everolimus and Voriconazole Interaction in Lung Transplant Patients.

BACKGROUND: To analyze the impact of voriconazole administration on everolimus dose, trough concentrations and concentration/dose (C0/D) ratio in order to determine the appropriate management of this interaction in lung transplant patients. METHODS: A retrospective study of 16 of consecutive lung transplant patients on a stable everolimus-based regimen to which oral voriconazole was added from January 2013 to February 2014. Everolimus blood levels were measured using the Thermo Scientific QMS Everolimus Immunoassay on an ARCHITECT-C8000 analyzer. The Wilcoxon signed-rank test was used to assess the exposure parameter variations before, during, and after azole withdrawal. A statistical analysis was performed using SPSS version 19.0. P-value < 0.05 was considered statistically significant. RESULTS: Sixteen patients were included. Voriconazole treatment led to a significant 8.7-fold increase in the everolimus C0/D ratio. Although initially the daily dose was reduced to 48.5% ± 20.5%, and subsequently to 79.5% ± 7.1%, the desired therapeutic levels were achieved in all patients when it was decreased to 86.6% ± 3.9%. After its withdrawal, the C0/D ratio returned to values similar to the baseline situation. The comparison of exposure parameters studied at stable moments, before and after the completion of azole treatment with the cotreatment period, revealed significant changes (P < 0.05). CONCLUSIONS: Oral voriconazole is a strong inhibitor of everolimus metabolism, requiring a dose reduction of around 87%. At the time of azole withdrawal, the dose should be increased to achieve C0/D ratio values similar to the initial situation. In our clinical practice, for a safe coadministration, a preemptive decrease to 75% of everolimus dose with the first azole prescription is recommended. Close monitoring of the everolimus concentrations and corresponding dosage adjustments are necessary until the target levels are achieved during both periods.

Patterns of drug therapy in newly diagnosed Spanish patients with systemic lupus erythematosus.

OBJECTIVES: This is the first Spanish multicentric inception lupus cohort, formed by SLE patients attending Spanish Internal Medicine Services since January 2009. We aimed to analyse drug therapy during the first year of follow-up according to disease severity. METHODS: 223 patients who had at least one year of follow-up were enrolled upon diagnosis of SLE. Therapy with prednisone, pulse methyl-prednisolone, hydroxychloroquine, immunosuppressives and calcium/vitamin D was analysed. RESULTS: Prednisone was given to 65% patients, at a mean (SD) daily dose of 11 (10) mg/d. 38% patients received average doses >7.5 mg/d during the first year. Patients with nephritis and with a SLEDAI ≥6 were treated with higher doses of prednisone. 81% of patients were treated with hydroxychloroquine, with higher frequency among those with a SLEDAI ≥6 (88% vs. 68%, p<0.001). The use of immunosuppressive drugs and methyl-prednisolone pulses was higher in patients with a baseline SLEDAI ≥6, however, differences were no longer significant when patients with lupus nephritis were excluded. The use of calcium/vitamin D increased with the dose of prednisone, however, 43% of patients on medium-high doses of prednisone did not take any calcium or vitamin D. CONCLUSIONS: This study gives a real-world view of the current therapeutic approach to early lupus in Spain. The generalised use of hydroxychloroquine is well consolidated. There is still a tendency to use prednisone at medium to high doses. Pulse methyl-prednisolone and immunosuppressive drugs were used in more severe cases, but not as steroid sparing agents. Vitamin D use was suboptimal.

Assessment of radiobiological metrics applied to patient-specific QA process of VMAT prostate treatments.

VMAT is a powerful technique to deliver hypofractionated prostate treatments. The lack of correlations between usual 2D pretreatment QA results and the clinical impact of possible mistakes has allowed the development of 3D verification systems. Dose determination on patient anatomy has provided clinical predictive capability to patient-specific QA process. Dose-volume metrics, as evaluation criteria, should be replaced or complemented by radiobiological indices. These metrics can be incorporated into individualized QA extracting the information for response parameters (gEUD, TCP, NTCP) from DVHs. The aim of this study is to assess the role of two 3D verification systems dealing with radiobiological metrics applied to a prostate VMAT QA program. Radiobiological calculations were performed for AAPM TG-166 test cases. Maximum differences were 9.3% for gEUD, -1.3% for TCP, and 5.3% for NTCP calculations. Gamma tests and DVH-based comparisons were carried out for both systems in order to assess their performance in 3D dose determination for prostate treatments (high-, intermediate-, and low-risk, as well as prostate bed patients). Mean gamma passing rates for all structures were bet-ter than 92.0% and 99.1% for both 2%/2 mm and 3%/3 mm criteria. Maximum discrepancies were (2.4% ± 0.8%) and (6.2% ± 1.3%) for targets and normal tis-sues, respectively. Values for gEUD, TCP, and NTCP were extracted from TPS and compared to the results obtained with the two systems. Three models were used for TCP calculations (Poisson, sigmoidal, and Niemierko) and two models for NTCP determinations (LKB and Niemierko). The maximum mean difference for gEUD calculations was (4.7% ± 1.3%); for TCP, the maximum discrepancy was (-2.4% ± 1.1%); and NTCP comparisons led to a maximum deviation of (1.5% ± 0.5%). The potential usefulness of biological metrics in patient-specific QA has been explored. Both systems have been successfully assessed as potential tools for evaluating the clinical outcome of a radiotherapy treatment in the scope of pretreatment QA.