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Summary: SAMHD1 is a dNTP triphosphohydrolase governing nucleotide pool homeostasis and can detoxify chemotherapy metabolites controlling their clinical responses. To understand SAMHD1 biology and investigate the potential of targeting SAMHD1 as neoadjuvant to current chemotherapies, we set out to discover selective small-molecule inhibitors. Here, we report a discovery pipeline encompassing a biochemical screening campaign and a set of complementary biochemical, biophysical, and cell-based readouts for rigorous characterization of the screen output. The identified small molecules, TH6342 and analogs, accompanied by inactive control TH7126, demonstrated specific, low μM potency against both physiological and oncology-drug-derived substrates. By coupling kinetic studies with thermal shift assays, we reveal the inhibitory mechanism of TH6342 and analogs, which engage pre-tetrameric SAMHD1 and deter oligomerization and allosteric activation without occupying nucleotide-binding pockets. Altogether, our study diversifies inhibitory modes against SAMHD1, and the discovery pipeline reported herein represents a thorough framework for future SAMHD1 inhibitor development.

Patients with recent pandemic coronavirus disease 19 (COVID-19) complain of neurological abnormalities in sensory functions such as smell and taste in the early stages of infection. Determining the cellular and molecular mechanism of sensory impairment is critical to understand the pathogenesis of clinical manifestations, as well as in setting therapeutic targets for sequelae and recurrence. The absence of studies utilizing proper models of human peripheral nerve hampers an understanding of COVID-19 pathogenesis. Here, we report that severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) directly infects human peripheral sensory neurons, leading to molecular pathogenesis for chemosensory impairments. An in vitro system utilizing human embryonic stem cell (hESC)-derived peripheral neurons was used to model the cellular and molecular pathologies responsible for symptoms that most COVID-19 patients experience early in infection or may develop as sequelae. Peripheral neurons differentiated from hESCs expressed viral entry factor ACE2, and were directly infected with SARS-CoV-2 via ACE2. Human peripheral neurons infected with SARS-CoV-2 exhibited impaired molecular features of chemosensory function associated with abnormalities in sensory neurons of the olfactory or gustatory organs. Our results provide new insights into the pathogenesis of chemosensory dysfunction in patients with COVID-19.

Background & Aims: One of the features of ulcerative colitis (UC) is a defect in the protective mucus layer. This has been attributed to a reduced number of goblet cells (GCs). However, it is not known whether abnormal GC mucus secretion also contributes to the reduced mucus layer. Our aims were to investigate whether GC secretion was abnormal in UC and exists as a long-term effect of chronic inflammation. Methods: Colonoids were established from intestinal stem cells of healthy subjects (HS) and patients with UC. Colonoids were maintained as undifferentiated (UD) or induced to differentiate (DF) and studied as three-dimensional or monolayers on Transwell filters. Total RNA was extracted for quantitative real-time polymerase chain reaction analysis. Carbachol and prostaglandin E2 mediated mucin stimulation was examined by MUC2 IF/confocal microscopy and transmission electron microscopy. Results: Colonoids from UC patients can be propagated over many passages; however, they exhibit a reduced rate of growth and transepithelial electrical resistance compared with HS. Differentiated UC colonoid monolayers form a thin and non-continuous mucus layer. UC colonoids have increased expression of secretory lineage markers ATOH1 and SPDEF, along with MUC2 positive GCs, but failed to secrete mucin in response to the cholinergic agonist carbachol and prostaglandin E2, which caused increased secretion in HS. Exposure to tumor necrosis factor α (5 days) reduced the number of GCs, with a greater percentage decrease in UC colonoids compared with HS. Conclusions: Chronic inflammation in UC causes long-term changes in GCs, leading to abnormal mucus secretion. This continued defect in GC mucus secretion may contribute to the recurrence in UC.

As computational methods are widely used in science disciplines, integrating computational thinking (CT) into classroom materials can create authentic science learning experiences for students. In this study, we classroom-tested a CT-integrated geoscience curriculum module designed for secondary students. The module consisted of three inquiry investigations that were age-appropriately translated from the computational practices of volcanologists who study tephra hazards and risks. We developed a domain-specific, block-code-based computational modeling environment where students carried out 1 of the 3 science practices in each inquiry investigation: experimentation, data visualization and interpretation, and modeling. We examined (1) student learning outcomes using pre- and post-tests and (2) student reflections on the computationally supported science practices surveyed at the end of each inquiry investigation during the module. Results indicate that students made statistically significant gains in science content as well as in computationally supported experimentation, data visualization and interpretation, and modeling practices. Over 80% of students identified different ways in which block coding supported their practices during inquiry investigations. Based on these findings, we discuss implications for the future development of computationally supported inquiry investigations in science.

Lentiviruses have been widely used as a means of transferring exogenous DNAs into human cells to treat various genetic diseases. Lentiviral vectors are fundamentally integrated into the host genome, but their integration sites are generally unpredictable, which may increase the uncertainty for their use in therapeutics. To determine the viral integration sites in the host genome, several PCR-based methods have been developed. However, the sensitivities of the PCR-based methods are highly dependent on the primer sequences, and optimized primer design is required for individual target sites. In order to address this issue, we developed an alternative method for genome-wide mapping of viral insertion sites, named CReVIS-seq (CRISPR-enhanced Viral Integration Site Sequencing). The method is based on the sequential steps: fragmentation of genomic DNAs, in vitro circularization, cleavage of target sequence in a CRISPR guide RNA-specific manner, high-throughput sequencing of the linearized DNA fragments in an unbiased manner, and identification of viral insertion sites via sequence analysis. By design, CReVIS-seq is not affected by biases that could be introduced during the target enrichment step via PCR amplification using site specific primers. Furthermore, we found that multiplexed CReVIS-seq, using collections of different single-guide RNAs (sgRNAs), enables simultaneous identification of multiple target sites and structural variations (i.e., circularized viral genome), in both single cell clones and heterogeneous cell populations.

Zinc finger protein with KRAB and SCAN domains 3 (ZKSCAN3) acts as an oncogenic transcription factor in human malignant tumors, including colon and prostate cancer. However, most of the ZKSCAN3-induced carcinogenic mechanisms remain unknown. In this study, we identified ZKSCAN3 as a downstream effector of the oncogenic Wnt/β-catenin signaling pathway, using RNA sequencing and ChIP analyses. Activation of the Wnt pathway by recombinant Wnt gene family proteins or the GSK inhibitor, CHIR 99021 upregulated ZKSCAN3 expression in a β-catenin-dependent manner. Furthermore, ZKSCAN3 upregulation suppressed the expression of the mitotic spindle checkpoint protein, Mitotic Arrest Deficient 2 Like 2 (MAD2L2) by inhibiting its promoter activity and eventually inducing chromosomal instability in colon cancer cells. Conversely, deletion or knockdown of ZKSCAN3 increased MAD2L2 expression and delayed cell cycle progression. In addition, ZKSCAN3 upregulation by oncogenic WNT/β-catenin signaling is an early event of the adenoma–carcinoma sequence in colon cancer development. Specifically, immunohistochemical studies (IHC) were performed using normal (NM), hyperplastic polyps (HPP), adenomas (AD), and adenocarcinomas (AC). Their IHC scores were considerably different (61.4 in NM; 88.4 in HPP; 189.6 in AD; 246.9 in AC). In conclusion, ZKSCAN3 could be responsible for WNT/β-catenin-induced chromosomal instability in colon cancer cells through the suppression of MAD2L2 expression.

Abstract The deoxycytidine analogue cytarabine (ara‐C) remains the backbone treatment of acute myeloid leukaemia (AML) as well as other haematological and lymphoid malignancies, but must be combined with other chemotherapeutics to achieve cure. Yet, the underlying mechanism dictating synergistic efficacy of combination chemotherapy remains largely unknown. The dNTPase SAMHD1, which regulates dNTP homoeostasis antagonistically to ribonucleotide reductase (RNR), limits ara‐C efficacy by hydrolysing the active triphosphate metabolite ara‐CTP. Here, we report that clinically used inhibitors of RNR, such as gemcitabine and hydroxyurea, overcome the SAMHD1‐mediated barrier to ara‐C efficacy in primary blasts and mouse models of AML, displaying SAMHD1‐dependent synergy with ara‐C. We present evidence that this is mediated by dNTP pool imbalances leading to allosteric reduction of SAMHD1 ara‐CTPase activity. Thus, SAMHD1 constitutes a novel biomarker for combination therapies of ara‐C and RNR inhibitors with immediate consequences for clinical practice to improve treatment of AML.

Introduction: Pharmacy practice has evolved to include direct patient care and interprofessional team models. Proper documentation of training and certification is required to verify eligibility for providing specialized services and for reimbursement. Objectives: The objectives of this study were to assess pharmacists’ views and perceptions on credentialing with respect to (1) familiarity and perceived importance of credentialing; (2) satisfaction with current credentialing tracking systems; and (3) challenges in adopting a centralized credentialing platform. Methods: This study used a cross-sectional, survey design to examine pharmacist perceptions of credentialing. The survey, distributed by the American Pharmacists Association from November 18, 2017 to December 2, 2017, consisted of 11 demographic items and 22 items about familiarity, importance, satisfaction and current systems of credentialing in pharmacy practice. Descriptive statistics were used to characterize the sample and outcome variables. Content analysis was conducted on freeform responses. Results: Data were analyzed from 446 (7.3%) completed surveys of the 6,144 distributed. Respondents were primarily represented by pharmacists from chain stores (29.6%), outpatient clinics (16.6%), and academic settings (15.2%). Job titles included staff pharmacist (33.9%), clinical pharmacist (21.3%), and manager positions (18.3%). Nine of 10 pharmacists reported familiarity with credentialing and considered credentialing as important to the pharmacy profession. Majority agreed with the importance of having a centralized online platform to store credentialing information (96.1%) and to obtain reimbursement (97.1%). Poor integration of data among different platforms (16%) was a common reason for dissatisfaction with current tracking systems. Most respondents (96.5%) were willing to provide information necessary for credentialing; however, over half were concerned about security of the information. Conclusions: This study was among the first to examine pharmacist perceptions of credentialing. Pharmacists in this study were familiar with and responsive to participating in credentialing process. They were also supportive of having a centralized credentialing system, but held reservations about security of information. Article Type: Original Research

Salt is one of the most important factors for fermented foods, but the effect of salt treatment time on the quality of fermented foods has rarely been studied. In this study, the effect of different salt treatment times (0, 48, and 96 h) after the start of fermentation on the quality of the soy sauce moromi extract (SSME) was investigated. As the salt treatment time was delayed, the population of Aspergillus oryzae, Lactobacillaceae, and Enterococcaecea in SSME increased, whereas the population of Staphylococcaceae and Bacillaceae decreased, leading to changes in the enzymatic activity and metabolite profiles. In particular, the contents of amino acids, peptides, volatile compounds, acidic compounds, sugars, and secondary metabolites were significantly affected by the salt treatment time, resulting in changes in the sensory quality and appearance of SSME. The correlation data showed that metabolites, bacterial population, and sensory parameters had strong positive or negative correlations with each other. Moreover, based on metabolomics analysis, the salt treatment-time-related SSME metabolomic pathway was proposed. Although further studies are needed to elucidate the salt treatment mechanism in fermented foods, our data can be useful to better understand the effect of salt treatment time on the quality of fermented foods.

Many citrus varieties are hybridized to improve their quality and to overcome the effects of climate change. However, there is limited information on the effect of the chemical profiles of hybrid varieties on their quality. In this study, we analyzed 10 citrus varieties and evaluated the correlation with their general characteristics and antioxidant activities. Chemical profiles, including the contents of sugars, organic acid compounds, flavonoids, limonoids, and carotenoids, which are related to taste, color, and health benefits, were significantly different depending on the citrus varieties, leading to different antioxidant capacities and general quality parameters. Based on these data, the correlations were investigated, and 10 citrus varieties were clustered into four groups—Changshou kumquat and Jeramon (cluster I); Setoka (cluster II-1); Natsumi, Satsuma mandarin, and Navel orange (cluster II-2); Kanpei, Tamnaneunbong, Saybyeolbong, and Shiranui (cluster II-3). Moreover, a metabolomic pathway was proposed. Although citrus peels were not analyzed and the sensory and functional qualities of the citrus varieties were not investigated in this study, our results are useful to better understand the relationship between citrus quality and metabolite profiles, which can provide basic information for the development and improvement of new citrus varieties.

We investigated the effect of agreements related to climate change on bilateral trade, using the Poisson pseudo-maximum likelihood (PPML) estimator on a large dataset that requires computing high-dimensional fixed effects. This study develops the concordance between commodities based on harmonized systems (HS) and technologies defined by both international and cooperative patent classifications (IPC/CPC). Using the OECD ENV-TECH classification, covering 169 exporting countries and 179 importing countries worldwide from 1991 to 2019, we determined the relationship between HS commodities and IPC/CPC climate change technologies. The main findings show that, first, the Kyoto Protocol has increased international bilateral trade in the climate change industry, while the Montreal Protocol has led to the opposite effect. In addition, the impact of environmental stringency on international bilateral trade in climate-related industries was negative and significant, refuting the Porter Hypothesis. This result was consistent within the trade between the convention countries. Therefore, national and industrial perspectives are relative, and voluntary climate mitigation methods should be applied, considering the areas where carbon reduction is relatively difficult. In other words, the conventions and policies must shift to a global paradigm in environmental protection and a more inclusive approach, with recognition of diversity in governance types.

Autophagy is an evolutionally conserved process that recycles aged or damaged intracellular components through a lysosome-dependent pathway. Although this multistep process is propagated in the cytoplasm by the orchestrated activity of the mTOR complex, phosphatidylinositol 3-kinase, and a set of autophagy-related proteins (ATGs), recent investigations have suggested that autophagy is tightly regulated by nuclear events. Thus, it is conceivable that the nucleolus, as a stress-sensing and -responding intranuclear organelle, plays a role in autophagy regulation, but much is unknown concerning the nucleolar controls in autophagy. In this report, we show a novel nucleolar–cytoplasmic axis that regulates the cytoplasmic autophagy process: nucleolar protein NOP53 regulates the autophagic flux through two divergent pathways, the ZKSCAN3-dependent and -independent pathways. In the ZKSCAN3-dependent pathway, NOP53 transcriptionally activates a master autophagy suppressor ZKSCAN3, thereby inhibiting MAP1LC3B/LC3B induction and autophagy propagation. In the ZKSCAN3-independent pathway, NOP53 physically interacts with histone H3 to dephosphorylate S10 of H3, which, in turn, transcriptionally downregulates the ATG7 and ATG12 expressions. Our results identify nucleolar protein NOP53 as an upstream regulator of the autophagy process.

This case study examines how material resistance (limitations posed by the physical world) and graph interpretation intersected during a high school biology investigation using digital sensors. We use an extended episode from a small group to illustrate how, in an inquiry-based unit, measuring near the resolution limit of a sensor caused scaling issues in graphs. Qualitative videotape analysis focuses on both the students' attempts to make sense of a perceived lack of variation in the collected data and the teacher's and classroom researchers' misinterpretation of the students' difficulties with graph interpretation. We suggest that these educators, though experienced, could have benefited from additional strategies to help them recognise and respond to graph interpretation issues introduced by digital representations of real-world data, and that the students could have benefited from explicit prompts to discuss the limitations of their equipment. We describe several implications for researchers, teachers, and curriculum developers interested in implementing inquiry-based biology investigations using sensor data. We argue that students should be supported to recognise that encountering unexpected results from their investigations and working to understand what these results have to say about the real world is an important and valid part of the practice of real science.

When integrated into online curriculum modules for students, educative curriculum materials (ECMs) can enhance teachers' enactment of these modules. This study investigated (1) the use of digitally enhanced ECMs built into an online plate tectonics curriculum module by teachers with different backgrounds and teaching experience, (2) the relationship between teachers' use of ECMs and student learning gains, and (3) teacher reflections on the value of the ECMs they used. We studied 26 teachers who taught middle and high school students (n = 1,098) by analyzing teacher log files automatically generated by the ECMs, teacher reflections collected with post-implementation surveys and interviews, student log files, and student learning gains from pretest to posttest. Results indicate that (1) there were large variations in the amounts and types of ECM features teachers accessed, (2) middle school teachers accessed significantly more ECM features than high school teachers p < 0.01, (3) students of teachers who used ECMs during class time made significantly higher learning gains than students of teachers who used them only before and/or between class time, p < 0.05, and (4) teachers most valued ECM features on student assessment. An overall non-significant, but positive, correlation between the total teacher interactions with ECMs and student learning gains was observed, r = 0.20, p = 0.32.

This study addresses the measurement of high school students' epistemic knowledge associated with scientific experimentation (EKSE) which concerns "how" scientific experimentation generates knowledge and "why" that knowledge is justified. Based on philosophical, educational standards, and literature analyses, an EKSE construct is characterized as (1) underlying students' decisions and reasoning elicited during experiment design, data collection/measurement, and data analysis/interpretation and (2) progressing in five levels of sophistication: no information, nascent, dogmatic, contextualized, and reflective. This study psychometrically examined the structure of the EKSE construct using the construct modeling approach. An instrument was designed to ask students decisions and reasoning encountered while carrying out the activities embedded in experimental inquiry. High school students took the instrument as a pretest and a posttest to a physics curriculum where they learned to carry out independent scientific experiments. Results indicate that (1) all decisions and reasoning taking place during experiment design, data collection/measurement, and data analysis/interpretation contribute to the EKSE construct, (2) epistemic knowledge develops in the hypothesized order, (3) scientifically aligned decisions are likely to occur when students' epistemic knowledge is at or higher than the dogmatic level, and (4) students' EKSE can improve after engaging in a curriculum that encourages independent scientific experimentation with materials. Implications of these results and further research are discussed.

This study investigates the microstructure and mechanical properties of wear-resistant hardfacing structures fabricated on an AISI 1045 carbon steel substrate using a specially designed Fe–Cr based powder for directed energy deposition (DED). Electron backscatter diffraction (EBSD) analysis reveals that the texture predominantly consists of cube rotated {001} and cube {001} textures, in addition to weaker Brass {112} texture components. These textures contribute to the random orientations of martensitic grains and facilitate the tracing of austenite reconstruction. Notably, the as-printed samples exhibited superior yield strength (999.4 ± 86.3 MPa) and ductility (9.6 ± 2.6%) along the build direction (BD), compared to conventional samples, which demonstrated a tensile strength of 790 MPa and ductility of 2%. This improvement is primarily attributed to the hardening effects associated with a low volume fraction of retained austenite and the precipitation of Cr carbides. Comprehensive mechanical response, nanoindentation hardness profile across the interface, and microstructural analyses were conducted to confirm the feasibility of using a Fe-based hardfacing alloy for DED. The findings underscore the outstanding balance of strength and ductility exhibited by as-printed hardfacing alloy, further enhanced by its high printability, highlighting its potential in producing wear-resistant structures for repair applications.

Abstract Effective enzyme stabilization through immobilization is essential for the functional usage of enzymatic reactions. We propose a new method for synthesizing elastic hydroxyapatite microgel (E-HAp-M) materials and immobilizing lipase using this mesoporous mineral via the ship-in-a-bottle-neck strategy. The physicochemical parameters of E-HAp-M were thoroughly studied, revealing that E-HAp-M provides efficient space for enzyme immobilization. As a model enzyme, lipase (LP) was entrapped and then cross-linked enzyme structure, preventing leaching from mesopores, resulting in highly active and stable LP/E-HAp-M composites. By comparing LP activity under different temperature and pH conditions, it was observed that the cross-linked LP exhibited improved thermal stability and pH resistance compared to the free enzyme. In addition, they demonstrated a 156% increase in catalytic activity compared with free LP in hydrolysis reactions at room temperature. The immobilized LP maintained 45% of its initial activity after 10 cycles of recycling and remained stable for over 160 days. This report presents the first demonstration of a stabilized cross-linked LP in E-HAp-M, suggesting its potential application in enzyme-catalyzed processes within biocatalysis technology.

Abstract T cell immunoglobulin and mucin-containing molecule 3 (TIM-3) exhibits unique, cell type- and context-dependent characteristics and functions. Here, we report that TIM-3 on myeloid cells plays essential roles in modulating lung inflammation. We found that myeloid cell-specific TIM-3 knock-in (FSF-TIM3/LysM-Cre+) mice have lower body weight and shorter lifespan than WT mice. Intriguingly, the lungs of FSF-TIM3/LysM-Cre+ mice display excessive inflammation and features of disease-associated pathology. We further revealed that galectin-3 levels are notably elevated in TIM-3-overexpressing lung-derived myeloid cells. Furthermore, both TIM-3 blockade and GB1107, a galectin-3 inhibitor, ameliorated lung inflammation in FSF-TIM3/LysM-Cre+/− mice. Using an LPS-induced lung inflammation model with myeloid cell-specific TIM-3 knock-out mice, we demonstrated the association of TIM-3 with both lung inflammation and galectin-3. Collectively, our findings suggest that myeloid TIM-3 is an important regulator in the lungs and that modulation of TIM-3 and galectin-3 could offer therapeutic benefits for inflammation-associated lung diseases.

The strategy of cultivating healthy fish by manipulating their gut microbiota composition using probiotics is crucial for establishing sustainable fish farms. Information on the gut microbiota composition of healthy fish is important for the efficient development of probiotics and the aquaculture strategies that would apply them. This study assessed the gut microbiota composition of farmed olive flounder according to the growth stage to provide useful information for the development of probiotics. The fish were classified according to their weight, and the intestines of five juveniles weighing less than 100 g and five subadults weighing between 400 and 800 g were used for analysis. The results of alpha diversity analysis confirmed higher richness and diversity in the subadult group than in the juvenile group. Beta diversity analysis revealed clear boundaries and distances between the groups; however, individuals within each group were similar. The two groups showed marked differences in their gut microbiota composition. Phylum-level analysis revealed that the most abundant phylum was Proteobacteria in the juvenile group and Firmicutes in the subadult group. Notably, genus-level analysis revealed that Vibrio accounted for more than 50% of the total composition in the juvenile group. Linear discriminant analysis effect size revealed that the genera Vibrio, Photobacterium, and Leuconostoc were representative of the juvenile group, whereas 19 genera, including Lactococcus and Vagococcus, were representative of the subadult group. Thus, information on the representative microorganisms abundantly present in the intestines of healthy olive flounder can be considered for the selection and development of a probiotic genus for olive flounder farming.

Abstract We study the gravitational edge mode in the N $$ \mathcal{N} $$ = 1 Jackiw-Teitelboim (JT) supergravity on the disk and its osp(2|1) BF formulation. We revisit the derivation of the finite-temperature Schwarzian action in the conformal gauge of the bosonic JT gravity through wiggling boundary and the frame fluctuation descriptions. Extending our method to N $$ \mathcal{N} $$ = 1 JT supergravity, we derive the finite-temperature super-Schwarzian action for the edge mode from both the wiggling boundary and the superframe field fluctuation. We emphasize the crucial role of the inversion of the super-Schwarzian derivative in elucidating the relation between the isometry and the OSp(2|1) gauging of the super-Schwarzian action. In osp(2|1) BF formulation, we discuss the asymptotic AdS condition. We employ the Iwasawa-like decomposition of OSp(2|1) group element to derive the super-Schwarzian action at finite temperature. We demonstrate that the OSp(2|1) gauging arises from inherent redundancy in the Iwasawa-like decomposition. We also discuss the path integral measure obtained from the Haar measure of OSp(2|1).

Background: /purposeIn cases where oral squamous cell carcinoma (OSCC) invades the jawbone, clinicians frequently observe abnormal attenuation on computed tomography (CT) and pathologic signal intensity (SI) on magnetic resonance (MR) imaging of the affected underlying bone marrow. This study introduced a concept of “underlying bone change” to examine its association with clinicopathological features and prognosis of OSCC, as well as its correlation with medullary invasion. Materials and methods: We enrolled 93 consecutive patients diagnosed with OSCC, who underwent mandibulectomy between 2010 and 2016. CT and MR images, along with electronic medical records, were reviewed to evaluate correlations between underlying bone changes, clinicopathological features, five-year overall survival, and medullary invasion. Results: Of the 93 patients, 69 (74.2%) exhibited underlying bone sclerosis on CT, and 74 (79.6%) displayed pathological SI on MR images. These underlying bone changes correlated with the T stage and recurrence, but not with overall survival.Medullary invasion, observed in 61 (65.6%) patients, was strongly associated with T and TNM stages and was linked to poorer overall survival.The underlying bone changes on CT and MR images were positively associated with medullary invasion; however, no significant differences were found in the occurrence of underlying bone changes between the subtypes based on the extent of medullary invasion. Conclusion: Underlying bone changes on CT and MR images can provide valuable insights into the aggressiveness of bone invasion by OSCC. Accurate interpretation of these imaging findings might be crucial for correctly delineating surgical margins and preventing the overestimation of tumor extent.

Surveillance for influenza deaths has been used to gauge the severity of influenza seasons. Traditional surveillance, which relies on medical records review and laboratory testing, might not be sustainable during a pandemic. We examined whether electronic death certificates might provide a surveillance alternative. We compared information retrieved from electronic death certificates that listed influenza (or a synonym) with information retrieved from medical charts on which influenza deaths were reported by traditional means in Los Angeles County, California, USA, during the 2009 influenza A(H1N1) pandemic and 2 subsequent influenza seasons. Electronic death certificate surveillance provided timely information, matched the demographics and epidemiologic curve of that obtained from traditional influenza-related death surveillance, and had a moderately positive predictive value. However, risk factors were underreported on death certificates. Because surveillance by electronic death certificates does not require obtaining and reviewing medical records, it requires fewer resources and is less burdensome on public health staff.

The discovery of potent therapeutic compounds against dengue virus is urgently needed. The NS2B-NS3 protease (NS2B-NS3pro) of dengue fever virus carries out all enzymatic activities needed for polyprotein processing and is considered to be amenable to antiviral inhibition by analogy. Virtual screening of 300,000 compounds using Autodock 3 on the GVSS platform was conducted to identify novel inhibitors against the NS2B-NS3pro. Thirty-six compounds were selected for in vitro assay against NS2B-NS3pro expressed in Pichia pastoris. Seven novel compounds were identified as inhibitors with IC50 values of 3.9 ± 0.6–86.7 ± 3.6 μM. Three strong NS2B-NS3pro inhibitors were further confirmed as competitive inhibitors with Ki values of 4.0 ± 0.4, 4.9 ± 0.3, and 3.4 ± 0.1 μM, respectively. Hydrophobic and hydrogen bond interactions between amino acid residues in the NS3pro active site with inhibition compounds were also identified.

IntroductionEmerging evidence highlights the potential advantages of the Mediterranean diet (MD) in preserving kidney function and slowing chronic kidney disease (CKD) progression. However, interventional studies on the MD are scarce in East Asian populations.MethodsThis randomized crossover trial aimed to assess the safety and short-term impact of the Mediterranean Proper Optimal Balance (MEDi-POB) diet in Korean patients with stage 3–4 CKD. Kidney function was assessed using the estimated glomerular filtration rate, which was calculated using the CKD Epidemiology Collaboration equation. Fifty patients with CKD were randomly assigned to two groups, each starting with a different 4-week intervention, followed by a 4-week washout period, followed by a switch to the other 4-week intervention. During the MEDi-POB intervention, patients received home delivery of meals twice daily, 5 days a week, while the control intervention comprised a conventional diet. Forty-six patients successfully completed the entire 12-week trial. Paired t-tests were conducted to assess mean differences between the two groups. A linear mixed model was used to adjust for sequence and period.ResultsDietary fat, fiber, and niacin intake were significantly higher following the MEDi-POB diet than following the control diet (p = 0.001 for fat, p < 0.001 for fiber, and p = 0.007 for niacin). The MEDi-POB diet also yielded slightly increased total CO2 levels (p = 0.043), indicating effective management of metabolic acidosis. Conversely, sodium and copper intake were significantly lower with the MEDi-POB diet (p = 0.032 and p = 0.037, respectively). Caloric intake increased, but body mass index slightly decreased from baseline after consuming the MEDi-POB diet. Dietary potassium intake exhibited a non-significant increase (p = 0.053), and no significant changes in serum (p = 0.883) and urine potassium levels (p = 0.087) occurred. Kidney function remained well-preserved following the MEDi-POB diet.ConclusionThese results indicate that the MEDi-POB diet is safe even in patients with advanced CKD, as it does not adversely affect serum and urine potassium levels and helps maintain kidney function.

Objectives: The present retrospective study reviewed the association among sarcopenia, myosteatosis, and overall survival (OS) in patients with postoperative colorectal cancer (CRC) with regard to age. Design: A retrospective study was conducted with a five-year follow-up. Setting: Data from all patients with CRC, who underwent surgery between February 2005 and April 2014, were reviewed. Participants: Data from 1053 patients (622 male [59.1%], 431 female [40.9%]; mean [± SD] age, 62.8 ± 11.8 years) were analyzed. Measurements: Patients were divided into three groups according to age: ≤50, 51–74, and ≥75 years. Data, including perioperative parameters, and the presence of sarcopenia and myosteatosis according to skeletal muscle index (SMI) and skeletal muscle radiodensity (SMD), respectively, were collected. Sarcopenia was evaluated using CT by calculating the SMI at the L3 level by dividing the area of the skeletal muscle by height squared (cm2/m2). SMD was also calculated using CT at the L3 level, but by evaluating fat attenuation according to Hounsfield units (HU). Results: Patient allocation according to age group was as follows: ≤50 years, n = 147 (14.0%); 51–74 years, n = 742 (70.5%); and ≥75 years, n = 164 (15.5%). The presence of sarcopenia and myosteatosis were statistically significant with increasing age (P = 0.004 and P

Zinc finger with KRAB and SCAN domain 3 (ZKSCAN3) upregulates genes encoding proteins involved in cell differentiation, proliferation and apoptosis. ZKSCAN3 has been reported to be overexpressed in several human cancers such as colorectal cancer and prostate cancer and is proposed as a candidate oncoprotein. However, the molecular mechanism by which ZKSCAN3 participates in carcinogenesis is largely unknown. Here, we evaluated ZKSCAN3 expression in uterine cervical cancers (CC) by immunohistochemistry using formalin-fixed, paraffin-embedded tissues from 126 biopsy samples from 126 patients. The clinicopathological findings were analyzed and compared with ZKSCAN3 expression levels. ZKSCAN3 was strongly overexpressed in CCs compared to adjacent non-neoplastic cervical mucosa tissues. Moreover, a gene copy number assay showed amplified ZKSCAN3 in CC samples. ZKSCAN3 overexpression was also significantly associated with poor overall survival of the patients. Overall, our findings indicate that ZKSCAN3 overexpression is a frequent event in uterine CC and is correlated with a poor clinical outcome. ZKSCAN3 could be developed as a molecular marker for prognostic prediction and early detection.

Appointment-based medication synchronization (ABMS) programs have been associated with increased adherence and persistence to chronic medications. Adherence to statin therapy, angiotensin-converting enzyme inhibitors (ACEIs), angiotensin receptor blockers (ARBs), and non-insulin antidiabetic medications (NIDM) are used to determine a health plan’s Centers for Medicare and Medicaid Services (CMS) Star Rating under a pay-for-performance model. The objective of this study was to evaluate the impact of implementing an ABMS program on overall pharmacy adherence measures for statins, ACEI/ARBs, and NIDM, as presented through the Electronic Quality Improvement Platform for Plans and Pharmacies (EQuIPP©) platform. This retrospective, pre-post ABMS program study evaluated EQuIPP© generated adherence performance measures, represented as proportion of days covered (PDC), 6-months before and 6- and 12-months after the ABMS service for statin therapy, ACEIs/ARBs, and NIDM. All adherence measures showed statistically significant improvement in PDC percentage post ABMS implementation, except for NIDM percentage in 6-months post-ABMS service. This study shows that a comprehensive medication synchronization program can enhance adherence measures that are important to health plans to increase CMS Star Rating under a pay-for-performance model.

Barriers to medication adherence stem from multiple factors. An effective and convenient tool is needed to identify these barriers so that clinicians can provide a tailored, patient-centered consultation with patients. The Modified Drug Adherence Work-up Tool (M-DRAW) was developed as a 13-item checklist questionnaire to identify barriers to medication adherence. The response scale was a 4-point Likert scale of frequency of occurrence (1 = never to 4 = often). The checklist was accompanied by a GUIDE that provided corresponding motivational interview-based intervention strategies for each identified barrier. The current pilot study examined the psychometric properties of the M-DRAW checklist (reliability, responsiveness and discriminant validity) in patients taking one or more prescription medication(s) for chronic conditions. A cross-sectional sample of 26 patients was recruited between December 2015 and March 2016 at an academic medical center pharmacy in Southern California. A priming question that assessed self-reported adherence was used to separate participants into the control group of 17 “adherers” (65.4%), and into the intervention group of nine “unintentional and intentional non-adherers” (34.6%). Comparable baseline characteristics were observed between the two groups. The M-DRAW checklist showed acceptable reliability (13 item; alpha = 0.74) for identifying factors and barriers leading to medication non-adherence. Discriminant validity of the tool and the priming question was established by the four-fold number of barriers to adherence identified within the self-selected intervention group compared to the control group (4.4 versus 1.2 barriers, p < 0.05). The current study did not investigate construct validity due to small sample size and challenges on follow-up with patients. Future testing of the tool will include construct validation.

Explaining phenomena associated with a system involves describing a system's structure and articulating the process through which the system's structure changes over time. This paper defines geo-sequential reasoning in the context of plate tectonics and uses it to analyse how students explain the geological processes that occur along convergent boundaries as part of the plate tectonics system. This study was part of design-based research on an online Plate tectonics module that included simulation-based modelling developed for secondary school students. We analysed students' explanations (n = 950) about phenomena found along a convergent boundary (1) as an oceanic plate and a continental plate move towards each other and (2) between two oceanic plates located on the opposite side of a tectonic plate from a divergent boundary. We also analysed images created by students of the simulation as evidence to support their explanations. We found that a majority of students used simulation-based evidence when describing the sequence of events along the convergent boundary and that the synced planet surface and cross-section views in the simulation supported students' inclusion of processes responsible for the events. These findings have implications for how teaching and research with dynamic simulations can support reasoning built with temporal evidence.

The present study constitutes the first attempt to identify low-temperature intestinal probiotics for olive flounder (Paralichthys olivaceus) and analyze their effects on this species at 20 °C, which is its optimum culture temperature. Initially, 291 intestinal bacteria were isolated, and Rummeliibacillus stabekisii PM227 (PM227), Microbacterium oxydans NB29 (NB29), and Microbacterium foliorum BZ24 (BZ24), which successfully passed the characterization tests, were considered for dietary supplementation. The formulated diets, denoted as Pro1, Pro2, and Pro3, contained 1 × 108 CFU/g of PM227, NB29, and BZ24, respectively, whereas the control diet contained no probiotics. Triplicates of 20 fish (10.34 ± 0.26 g) were randomly allocated to the four diet groups and fed one of the diets for 8 weeks. At the end of the trial, Pro2 and Pro3 significantly increased growth and feed utilization parameters compared to the control and Pro1 groups. Only Pro2 and Pro1 improved (p < 0.05) myeloperoxidase activity and the transcription of interleukin (IL)-10 in the fish liver, respectively. Amylase, lipase, and protease activities were increased in the flounder intestine in all three potential probiotics groups, except for amylase in Pro2. Intestinal microbial richness (Chao1 index) and diversity (Shannon index) were higher in Pro2 than in the other diet groups, with some similarities observed between Pro1 and Pro3. A significantly lower relative abundance of Firmicutes and Staphylococcales was found in the intestine of all probiotics-supplemented flounder. Moreover, the relative abundance of Lactobacillus and Staphylococcus exhibited the following order among the examined groups: Pro1 > Pro3 > Pro2 > control and control > Pro2 > Pro1 > Pro3, respectively. Therefore, the identified low-temperature potential probiotics increased growth, immunity, digestive enzyme activities, and intestinal microbial populations in olive flounder, demonstrating their value as feed supplements in the flounder industry.

BACKGROUND: Despite the endemic nature of Echinococcus granulosus and Echinococcus multilocularis infection in regions of the United States (US), there is a lack of data on echinococcosis-related mortality. To measure echinococcosis-associated mortality in the US and assess possible racial/ethnic disparities, we reviewed national-death certificate data for an 18-year period. METHODOLOGY/PRINCIPAL FINDINGS: Echinococcosis-associated deaths from 1990 through 2007 were identified from multiple-cause-coded death records and were combined with US census data to calculate mortality rates. A total of 41 echinococcosis-associated deaths occurred over the 18-year study period. Mortality rates were highest in males, Native Americans, Asians/Pacific Islanders, Hispanics and persons 75 years of age and older. Almost a quarter of fatal echinococcosis-related cases occurred in residents of California. Foreign-born persons accounted for the majority of echinococcosis-related deaths; however, both of the fatalities in Native Americans and almost half of the deaths in whites were among US-born individuals. CONCLUSIONS/SIGNIFICANCE: Although uncommon, echinococcosis-related deaths occur in the US. Clinicians should be aware of the diagnosis, particularly in foreign-born patients from Echinococcus endemic areas, and should consider tropical infectious disease consultation early.

Fruit and Vegetable Prescription (FVRx) Programs combine produce prescriptions and nutrition education to reduce fruit and vegetable consumption barriers and promote health among low-income patient populations. This study examined whether a multi-level FVRx intervention model with intensive education improves dietary behaviors, food security, and health outcomes over single-level interventions alone. A 6-month nonrandomized, parallel, controlled trial was conducted with one intervention, FVRx (n = 31) and two comparison groups, Ad hoc Nutrition Education (n = 13) and Control (n = 16). The FVRx group received produce prescriptions (US$1/day/household member) redeemable at a farmer's market, two SNAP-Ed programs, one financial literacy program, and monthly health screenings. The Nutrition Education (NE) group participated in one SNAP-Ed program, and the Control group received safety-net clinic care only. Surveys assessed dietary intake, food security, food purchasing practices, and financial and food resource management. Pre-post clinical biomarkers (blood lipid and hemoglobin A1c) and monthly biometrics (anthropometrics and blood pressure) were measured. Descriptive analysis and one-way analysis of variance (ANOVA) were conducted. Compared with comparison groups, FVRx participants significantly increased the frequency of consuming dark green vegetables, FVRx (0.36 ± 0.72); NE (0.14 ± 0.33); Control (-0.09 ± 0.19) cups/day (p < 0.05). FVRx participants significantly improved multiple healthful food purchasing practices, and the ability to afford more utilities (FVRx (33%); NE (0%); Control (10%); p < 0.05). Limited changes were observed in food security and clinical biomarker outcomes between groups. Combining expanded nutrition and financial literacy education with produce prescriptions improves low-income adults' financial and food resources, preference, knowledge, purchase, and consumption of locally grown vegetables over single-level interventions.